Mosquito Management Plan and Environmental Assessment

Home , Berosus exiguus, Brachydeutera, Brachydeutera argentata, Crossopriza lyoni, Culiseta inornata, Culiseta melanura

DRAFT Mosquito Management Plan and Environmental Assessment

for the

Great Meadows Unit at the Stewart B. McKinney National Wildlife Refuge

Prepared by: ______Date:______Refuge Manager

Concurrence:______Date:______Regional IPM Coordinator

Concured:______Date:______Project Leader

Approved:______Date:______Assistant Regional Director Refuges, Northeast Region

Table of Contents

Chapter 1 PURPOSE AND NEED FOR PROPOSED ACTION ...... 5 1.1 Introduction ...... 5 1.2 Refuge Location and Site Description ...... 5 1.3 Proposed Action ...... 7 1.3.1 Purpose and Need for Proposed Action ...... 7 1.3.2 Historical Perspective of Need ...... 9 1.3.3 Historical Mosquito Production Areas of the Refuge ...... 10 1.3.4 How the Proposed Action Would Be Accomplished ...... 10 1.3.5 Objectives of the Proposed Action ...... 11 1.4 Issues and Concern ...... 11 1.4.1 Public Participation ...... 11 1.4.2 Issues Related to the Proposed Action ...... 11 1.5 Summary of Laws, Regulations, and Policies Governing the Proposed Action ...... 11 1.5.1 National Wildlife Refuge System Administration Act of 1966 ...... 11 1.5.2 National Wildlife Refuge System Regulations ...... 12 1.5.3 Endangered Species Act (ESA) of 1973, as amended (16 U.S.C.1531-1544) ...... 12 1.5.4 Biological Integrity, Diversity, and Environmental Health policy (BIDEH, 601 FW 3) ...... 12 1.5.5 Integrated Pest Management (IPM) policy (569 FW 1) ...... 12 1.5.6 Appropriate Refuge Uses policy (603 FW 1) ...... 13 1.5.7 Compatibility policy (603 FW 2) ...... 13 1.5.8 Special Use Permits ...... 13 1.5.9 National Environmental Policy Act (NEPA) of 1969, as amended (42 U.S.C. 4321-4347) ...... 14 1.6 Decision to be Made ...... 14

Chapter 2 ALTERNATIVES INCLUDING THE PROPOSED ACTION ...... 15 2.1 The Process Used to Develop the Alternatives ...... 15 2.2 Description of Alternatives, Including the Proposed Action and No Action ...... 15 2.2.1 Factors Common to All Alternatives ...... 15 2.3 Alternative A – Proposed Action – Phased Approach to Mosquito Control ...... 15 2.3.1 Phase 1 ...... 18 2.3.2 Phase 2 ...... 18 2.3.3 Phase 3 ...... 19 2.3.4 Phase 4 ...... 19 2.3.5 Phase 5 ...... 19 2.3.6 Access ...... 20 2.3.7 Mosquito Monitoring and Surveillance ...... 20 2.3.8 Wildlife Monitoring and Surveillance ...... 21 2.3.9 Pesticide Approval Process ...... 21 2.3.10 Mosquito Control Pesticides ...... 22 2.3.11 Annual Meeting/Training ...... 25 2.4 Alternative B – Monitoring and Larvicide Only ...... 26 2.5 Alternative C – No Action ...... 26 2.6 Alternatives Summary ...... 27

Chapter 3 AFFECTED ENVIRONMENT ...... 27 3.1 Physical Environment ...... 27 3.1.1 Climate ...... 27 3.1.2 Topography, Geology, and Soils ...... 28 3.1.3 Air Quality ...... 28 3.1.4 Water and Water Quality ...... 29 3.2 Biological Considerations ...... 29 3.2.1 Vegetation and Habitat Types ...... 29

3.2.2 Special Biological Communities/Critical Wildlife Habitat ...... 30 3.2.3 Noxious Weeds/Exotic Plants ...... 30 3.2.4 Wildlife ...... 34 3.3 Socioeconomic Considerations ...... 40 3.3.1 Cultural Resources ...... 40 3.3.2 Socioeconomics and Environmental Justice...... 40 3.3.3 Land Use ...... 41 3.3.4 Aesthetics ...... 42

Chapter 4 ENVIRONMENTAL CONSEQUENCES ...... 42 4.1 Impacts on the Physical Environment ...... 43 4.1.1 Impacts to Air Quality...... 43 4.1.2 Impacts to Topography...... 44 4.1.3 Impacts to Water ...... 44 4.2 Impacts on Biological Resources ...... 46 4.2.1 Impacts to Vegetation ...... 46 4.2.2 Impacts on Mammals...... 47 4.2.3 Impacts on Birds...... 48 4.2.4 Impacts on Reptiles and Amphibians ...... 51 4.2.5 Impacts on Fisheries ...... 52 4.2.6 Impacts on Invertebrates ...... 53 4.3 Impacts on the Social Environment ...... 55 4.3.1 Impacts on Cultural and Historic Resources ...... 55 4.3.2 Impacts on Land Use ...... 55 4.3.3 Impacts on Human Health and Safety Concerns ...... 55 4.4 Cumulative Impacts ...... 56 4.4.1 Alternative A – Proposed Action ...... 56 4.4.2 Alternative B – Monitoring and Larvacides...... 56 4.4.3 Alternative C - No Action...... 56

Chapter 5 COMPLIANCE, CONSULTATION, AND COORDINATION WITH OTHERS ...... 57 5.1 Agency Coordination and Public Involvement ...... 57 5.2 Environmental Review and Consultation ...... 57 5.3 Other Federal Laws, Regulations, and Executive Orders...... 57 5.4 Distribution and Availability ...... 57

LITERATURE CITED ...... 58

TABLES Table 1. Phased-response mosquito management guidelines for the Great Meadows Unit at the Stewart B. McKinney NWR ...... 17 Table 2. Common pesticides used for mosquito control in wetlands of Connecticut ...... 22 Table 3. Summary of mosquito management activities permitted under each alternative ...... 27 Table 4. Federal and State listed wildlife and plant species that occur or have the potential to occur on the Stewart B. McKinney NWR ...... 31 Table 5. Finfish species found in the waters near the Great Meadows Unit of SBMNWR ...... 38 Table 6. Summary of biological impacts of the pesticides used for mosquito control in wetlands by the CT MMP ...... 48

FIGURES Figure 1. Map of the entire Refuge showing all units taken from the draft CCP maps...... 6 Figure 2. Map of the Great Meadows Unit taken from the draft CCP maps...... 7

APPENDICES APPENDIX A. USFWS Director’s Memorandum dated May 27, 2014, Mosquito Management on National Wildlife Refuges APPENDIX B. CT MMP – Mosquito Management an Integrated Approach APPENDIX C. USFWS Compatibility Determination APPENDIX D. USFWS Pesticide Use Proposal – Example Form APPENDIX E. USFWS Special Use Permit – Example Application APPENDIX F. Connecticut MMP’s West Nile Virus Surveillance & Response Plan, Contingency Plan for Eastern Equine Encephalitis, and Zika Virus Surveillance and Response Plan APPENDIX G. CT Trapping and Arbovirus Testing Program APPENDIX H. Effects of Larvicides on Non-Target Organisms APPENDIX I. Environmental Effects of Mosquito Control APPENDIX J. Mosquito Biology APPENDIX K. Pyrethrins and Pyrethroids APPENDIX L. Summary of Connecticut Mosquito Testing from 2016 APPENDIX M. Public Comment and Response

Note: Appendices have their own page numbers.

Chapter 1 PURPOSE AND NEED FOR PROPOSED ACTION

1.1 Introduction The U.S. Fish and Wildlife Service (Service) considers mosquitoes a part of the natural ecosystem in most National Wildlife Refuge System (Refuge System) habitats in which they occur. We allow mosquitoes on refuges to exist unimpeded unless they pose a specific human risk, or in some unique locations (e.g., Hawaii) a wildlife health risk. A Refuge Manager may authorize others to conduct mosquito management activities on a refuge to protect public health when local, current mosquito monitoring data provided by the public health agency or an authorized designated representative indicate that mosquitoes on the refuge are causing, or are expected to cause, a public health threat.

Unless mosquitoes interfere with refuge-specific management goals and objectives, or cause a public health risk, they are allowed to exist unimpeded on a refuge. Mosquito-vectored pathogens that cause disease are the primary public health concern associated with mosquitoes on a refuge. When faced with mosquito management decisions affecting Refuge System lands and waters, our position is to work with others to protect the public health using the most effective method or combination of methods that pose the lowest risk to fish, wildlife, and their habitats. All Refuge System mosquito management activities, including Service planning documents, must be consistent with all applicable Federal laws, regulations and policies.

The management of mosquitoes in Connecticut is a collaborative effort involving the Department of Energy & Environmental Protection (DEEP), the Connecticut Agricultural Experiment Station (CAES) and the Department of Public Health (DPH), together with the Department of Agriculture and the Department of Pathobiology at the University of Connecticut (UCONN). These agencies are responsible for monitoring and managing the state’s mosquito population levels to reduce the potential public health threat of mosquito-borne diseases. The program was created by a legislative act (PA 97-289) in 1997 to monitor and control the spread of eastern equine encephalitis (EEE), a potentially deadly disease. In 1999, West Nile virus (WNV) was discovered in New York, New Jersey, and Connecticut. Although WNV causes fatal illness in a smaller proportion of cases than EEE, its greater potential to cause large outbreaks makes it an important health concern. Currently, the Connecticut Agricultural Experiment Station (CAES) maintains a network of 91 fixed mosquito trapping stations located in 72 municipalities throughout the state.

From 1998 until 2007, the Connecticut Mosquito Management Program (MMP) conducted surveillance and applied larvicide on less than 5 acres of the Great Meadow Unit of the Stewart B. McKinney National Wildlife Refuge (see figure 2; NWR; Refuge). Beginning in 2003 a marsh restoration project was conducted at the Great Meadows Unit, resulting in decreased populations of mosquitoes in restored sites. Following the restoration project, there was a decline in the number of locations identified as areas of public health concern from 9 sites to 2 (see changes between figure 2 and 3 in Appendix C). In 2008, surveillance and treatment of the Great Meadow Unit was suspended pending review of the U.S. Fish and Wildlife Service’s new compatibility process (Wolfe 2015 personal communication; see Appendix C for CD). However, the CT DEEP continues to identify specific areas of public health concern in the Great Meadows Unit on an annual basis.

1.2 Refuge Location and Site Description Stewart B. McKinney NWR is comprised of 10 units stretched across 70 miles of Connecticut's coastline (see figure 1). It was established in 1972 and was originally called Salt Meadow National Wildlife Refuge. The Refuge was renamed in 1987 to honor the late U.S. Congressman Stewart B. McKinney, who was instrumental in expanding it. The purposes for this Refuge are: • ... for use as an inviolate sanctuary, or for any other management purpose, for migratory birds (16 U.S.C. § 715 et seq.). • To enhance the populations of herons, egrets, terns, and other shore and wading birds within the Refuge; • To encourage natural diversity of fish and wildlife species within the Refuge; • To provide for the conservation of all fish and wildlife within the Refuge; • To fulfill the international treaty obligations of the United States respecting fish and wildlife; and

• To provide opportunities for scientific research, environmental education, and fish and wildlife- orientated recreation. • ... for the development, advancement, management, conservation, and protection of fish and wildlife resources ... (16 U.S.C. § 742f(a)(4)) • ... for the benefit of the United States Fish and Wildlife Service, in performing its activities and services. Such acceptance may be subject to the terms of any restrictive or affirmative covenant, or condition of servitude ... (16 U.S.C. § 742f(b)(1))

Figure 1: Map of the entire Refuge showing all units taken from the draft CCP maps.

Figure 2: Map of the Great Meadows Unit taken from the draft CCP maps. Refuge owned boundary is in YELLOW. Acquisition boundary is in ORANGE.

1.3 Proposed Action The Refuge proposes to implement an Integrated Pest Management Plan (IPM) that consists of a phased approach to mosquito management on the Great Meadows Unit of the Stewart B. McKinney NWR. The IPM includes ongoing coordination with the Connecticut Mosquito Management Program (MMP) and adheres to U.S. Fish and Wildlife Service (Service; USFWS) regulations and policies for mosquito-management.

1.3.1 Purpose and Need for Proposed Action The purpose of the Proposed Action is to ensure that activities to survey and control mosquito populations on the Great Meadows Unit of the Refuge are compatible with the establishing purposes of the Refuge. With the spread of West Nile Virus (WNV) and the potential for other mosquito-borne disease to spread, there is increasing pressure to manage mosquito populations that occur on lands of the NWRS, especially in densely populated areas like the Connecticut coast. The Refuge considers mosquitoes a natural component of tidal wetlands but also recognizes that mosquitoes may pose a threat to human health. Mosquito control management plans, such as this one, and documentation of management actions on refuges are necessary to protect biological integrity and ecological health, as well as sensitive species and to ensure the health and welfare of surrounding human populations.

The Service uses regulations and policies to plan and guide mosquito management actions on refuges. Enabling regulations of the Refuge System are contained in Title 50 Code of Federal Regulations (CFR) Subchapter C, Part 25, Administrative Provisions, as well as the National Wildlife Refuge System Administration Act of 1966, as amended by the National Wildlife Refuge System Improvement Act of 1997, as amended (16 U.S.C. §§ 668dd- 668ee). Guiding policies for mosquito management are referenced in a USFWS Director’s Memorandum dated May 27, 2014 (see Appendix A) and include the: • Comprehensive Conservation Planning Process (602 FW 3) • Step-Down Management Planning (602 FW 4) • Biological Integrity, Diversity, and Environmental Health (601 FW3) 7

• Integrated Pest Management (569 FW 1) • Appropriate Refuge Uses (603 FW 1) • Compatibility (603 FW 2)

Mosquitoes (members of the Phylum Arthropoda) are well known vectors of disease to both humans and wildlife and in rare cases can cause serious illness and death. “Arthropod-borne viruses (termed "arboviruses") are viruses that are maintained in nature through biological transmission between susceptible vertebrate hosts by blood-feeding arthropods (mosquitoes, sand flies, ceratopogonids "no-see’ems", and ticks). Vertebrates can become infected when an infected arthropod bites them to take a blood meal” (CDC 2010). Recently, the arbovirus labeled West Nile virus (WNV) has been of particular concern across the United States and on the coast of Connecticut. As a result, Service personnel across the refuge system have undertaken a number of actions, including: stepping-up coordination and communication with mosquito experts at state agencies, universities, and elsewhere; increasing communication with public health officials; participating in mosquito management seminars and workshops; initiating mosquito management-oriented research on refuges; and conducting restoration that benefits natural resources and reduces the need for pesticide application to control mosquito populations.

The association between mosquitoes and certain vector-borne diseases is also well known (e.g., West Nile virus). Historic measures for reducing mosquitoes included draining or filling wetlands, but today, a suite of other measures are now in place and include the placement and maintenance of mosquito ditches and application of pesticides (Resh 2001). The placement and maintenance of ditches in salt marshes is a physical technique that was used throughout salt marshes in the northeast to reduce mosquito production where water collects in depressions. The purpose of ditching is to increase tidal flushing and permit access by fish that can predate upon mosquito larvae (Resh 2001). Ditches were placed throughout the Refuge over the last century but results are typically short-lived (e.g., 10 years). Today the Refuge and the CT MMP advocate for an integrated approach to mosquito management that includes a range of tools to improve habitat conditions for estuarine wildlife while reducing threats to public health from mosquito species capable of transmitting disease to humans.

1.3.1a West Nile Virus. In the United States, WNV is transmitted by infected mosquitoes, primarily members of the Culex and Aedes species, although 64 species have been identified in WNV positive mosquito pools in the United States since 1999 (CDC, June 2010). The WNV has been isolated from 18 different species of mosquitoes in Connecticut, but 5 species have been implicated as the most important vectors: Cx. pipiens, Cx. restuans, Cx. salinarius, Culiseta melanura, and Aedes vexans. Cx. pipiens, Cx. Restuans, and Cs. melanura principally feed on birds and are largely involved in perpetuating the virus among wild bird populations in nature, while Cx. salinarius and Ae. vexans readily feed on mammals including humans and are believed to be involved in transmission of WNV to horses and humans. Cx. pipiens, a peridomestic species that develops in water with high organic content and is particularly abundant in urban centers, is the most frequently infected mosquito species in the state (CT DEEP MMP website). A host of arbovirus mosquito vector species were found or have the potential to be found on the Refuge. A summary of the CT MMP’s surveillance from 2003-2007 lists 12 of these species, which disease it can carry, the flight range, and whether it is known to occur on the Refuge (see Appendix C – table 3). There were 69 cases of WNV in the State from 2000-2009; 32 of those from Fairfield County where the Great Meadows Unit is located and 51 cases of WNV from 2010-2014 – 32 of those were from Fairfield County. There have been 3 fatal incidents of WNV in the State from 2000-2014 (CT Department of Public Health website).

1.3.1b Eastern equine encephalitis. This is a rare but serious disease caused by a virus that is spread by adult mosquitoes. On average there are 5 cases each year in the United States. There has never been a documented human case of EEE in Connecticut, but the virus is found in birds and bird-biting mosquitoes that live near wetland habitats along the eastern seaboard from New England to Florida. In some years high numbers of birds are infected with EEE, which could increase the likelihood that the types of mosquitoes that bite both mammals and birds will become infected. These mosquitoes can then infect people and horses. EEE is not spread directly by people and horses with the disease. The risk of getting EEE is highest from late July through September. 8

The virus responsible for EEE attacks the central nervous system of its host. Horses are particularly susceptible to the infection and mortality rates approach 100%. Onset is abrupt and EEE in horses is almost always fatal. Signs of the disease in horses include unsteadiness, erratic behavior, loss of coordination and seizures. There is no effective treatment for the disease and death can occur within 48 to 72 hours of the horse's first indications of illness. Horses can and should be inoculated against this disease, especially in areas where EEE is known to circulate.

In humans, symptoms of EEE appear from 3 to 10 days after being bitten by an infected mosquito. Some infected people may not develop illness. For those who become ill, the clinical symptoms may include high fever (103 to 106 degrees F), stiff neck, headache, and lack of energy. Inflammation of the brain, encephalitis, is the most dangerous of the clinical symptoms. The disease gets worse quickly and some patients go into a coma within a week. Once symptoms develop, treatment for EEE is supportive and aimed at reducing the severity of the symptoms. As many as one-third of people who get the disease die from it and of those who survive, approximately one-half will have permanent neurologic damage. Presently, there is no available vaccine for use in humans (CT MMP Website). One case of EEE was reported in Connecticut from 2004-2013 (CDC Website EEEV).

1.3.2 Historical Perspective of Need The first record of mosquito transmitted diseases to rise to epidemic level in the colony of Connecticut occurred with an outbreak of Yellow Fever in New Haven in 1743. An outbreak of Yellow Fever in June 1794 claimed 64 citizens of New Haven; 150 people were stricken with the disease (Anderson 2010). Seven Yellow Fever epidemics were recorded between 1743 and 1820. Yellow fever is an acute infectious disease of short duration and extremely variable severity, caused by a virus that is transmitted by mosquito bite and followed by life-long immunity in the survivors (Kerr 1951).

Malaria was another mosquito transmitted disease known to the colony of Connecticut spanning from the late 1600s to the early 1800s with the outbreaks rising to epidemic levels. The Connecticut State Board of Health reported that between 1894 and 1903, 1,073 citizens of the state of Connecticut died due to Malaria. Symptoms of Malaria are high fever, chills, and rigger, which can lead to death. (Anderson 2010).

It was not until the late 1800s and early 1900s when it was determined the route of transmission of Yellow Fever and Malaria was from the bite of a mosquito. Armed with this knowledge the State and Local governments began projects to control mosquitos to curb the transmission of mosquito-related diseases (Anderson 2010).

Between 1903 and 1915 Wilton Britton, the Connecticut State Entomologist, studied the State salt marsh and mosquitos to determine possible effects mosquito populations have on public health. A result from this study was the development of a mosquito control plan, which called for the drainage of salt and fresh water marshes to control mosquito populations. The control procedure also called for the use of oil as an asphyxiant to control mosquito production until the marsh could be drained (Anderson 2010).

In 1933 the Federal Government provided funding through the Civil Works Administration to control mosquito populations. The State of Connecticut utilized this funding to drain marshes and install and maintain tide gates and dikes to reduce mosquito populations. Starting in 1936 federal funding for mosquito control came from the Works Progress Administration, which continued for the State until 1940 when it was severely reduced. During this time period, the area that would become the Salt Meadow and Great Meadow Units of the Refuge were ditched to help control mosquitos (the Great Meadow area was ditched by the land owner to control mosquitoes).

In 1945 the use of pesticides began in the State. The first experimental use of pesticides was DDT, which was used to control mosquitos at a series of outdoor concerts for the comfort of concert attendees and not for public health reasons. From that date to the present time, pesticides have been used as a part of mosquito population management in Connecticut (Anderson 2010).

In 1999 WNV was discovered in New York, New Jersey, and Connecticut. The outbreak, in which seven humans and six horses died in New York and hundreds of birds died within the three states, was the first documented infection of WNV in the Western Hemisphere. Unlike EEE, WNV is new to the Americas and native birds have not developed a natural 9

immunity to the virus. Hence, a large proportion of the birds bitten by WNV-infected mosquitoes died. Historically, sporadic outbreaks of WNV have occurred in parts of Africa and Eurasia since 1937. The virus is similar to the virus that causes St. Louis Encephalitis (SLE) and causes similar symptoms in humans. Although WNV causes fatal illnesses in a smaller proportion of cases than EEE, its greater potential to cause large outbreaks makes it an important health concern.

The management of mosquitoes in Connecticut is a collaborative effort involving the Department of Energy & Environmental Protection (DEEP), the Connecticut Agricultural Experiment Station (CAES) and the Department of Public Health (DPH), together with the Department of Agriculture and the Department of Pathobiology at the University of Connecticut (UCONN). These agencies are responsible for monitoring and managing the state’s mosquito population levels to reduce the potential public health threat of mosquito-borne diseases. The program was created by a legislative act (PA 97-289) in 1997 to monitor and control the spread of EEE, a potentially deadly disease. The EEE is a virus that is present in nature and is cycled in the wild bird population by certain species of bird-feeding mosquitoes. The virus has no effect on wild birds; however, it can be fatal to humans, horses, and commercial exotic fowl (e.g., pheasants, emus). In Connecticut, outbreaks of EEE have occurred sporadically among horses and domestic pheasants since 1938, but no human cases have ever been confirmed. However, human deaths have occurred in nearby states.

1.3.3 Historical Mosquito Production Areas of the Refuge As part of the statewide MMP, the CT DEEP has previously been allowed to monitor and control larval mosquito populations on the Refuge, at both the Salt Meadow Unit in Westbrook and the Great Meadows Unit in Stratford. In the early 1990s, CT DEEP performed Open Marsh Water Management in the mosquito producing areas of the Salt Meadow Unit, essentially eliminating salt marsh mosquito producing sites there. As part of their WNV surveillance and response planning CT DEEP began monitoring at Great Meadows Unit and Salt Marsh Unit in 1998.

There were 9 sites at Great Meadows that were monitored, surveyed, and treated from 1998-2002. They were labelled Site A through Site I. However in 2003, CT DEEP assisted in the restoration of 40 acres of tidal wetlands as part of the Stratford Development Company mitigation project. In doing so, many of the mosquito producing sites that were once part of CT DEEP’s larviciding program were eliminated. Two sites were created from the old sites (sites D & G) totaling about 5 acres of marsh. These sites are the remaining mosquito production areas on the Refuge according to the CT MMP’s monitoring and surveillance. There have been discussions within the last few years between the Service, CT DEEP, and National Oceanic and Atmospheric Administration (NOAA Fisheries) regarding additional tidal wetland restoration on the Refuge in completion with the Lordship/ Raymark Restoration Plan/ Environmental Assessment. Like the previous restoration work done in 2003, this restoration project should reduce or eliminate many of the salt marsh mosquito producing areas on the Refuge. In the interim, these sites continue to produce large numbers of mosquitoes.

1.3.4 How the Proposed Action Would Be Accomplished The Proposed Action provides a phased approach for surveillance, monitoring, and control of mosquitoes on the Great Meadows Unit of the Refuge in a manner consistent with Federal laws, regulations and policies on National Wildlife Refuges), State response plans for mosquito borne diseases, and in partnership with the CT MMP. The proposed action serves as the Refuge’s Mosquito Management Plan (Plan). A Compatibility Determination for mosquito management activities is included in Appendix C. The Refuge has made an initial determination that the proposed plan is appropriate and compatible with Refuge purposes, pending input by the public.

Each year the Refuge will work with the CT MMP to develop the Special Use Permit (SUP) that will cover the surveillance, monitoring, and control activities allowed on the Refuge that year. An annual meeting between the Refuge and the CT MMP will ensure that permits are current, communication is continuous, and concerns related to mosquito populations and other biological resources of the Refuge are addressed. Vital to the mission of our respective agencies is maintaining a positive and productive working relationship. Pesticide Use Permits (PUP) and Pesticide Use Reports will be prepared annually by Refuge staff with data support from the CT MMP. In addition, prior to issuing the SUP, we will review the Section 7 consultation, cultural resource compliance, and this Environmental Assessment to determine if any additional documentation will be necessary.

1.3.5 Objectives of the Proposed Action • Protect public from mosquito-borne diseases • Protect threatened and endangered species, migratory birds, and other wildlife and their habitats from mosquito- borne diseases • Allow compatible surveillance of mosquito populations on the Refuge Development of Refuge-based phased response plan • Where mosquito control is needed based on established thresholds and survelillance data, use the most effective means that pose the lowest risk to wildlife and associated habitats • In rare cases, if other control methods have not been successful, allow the use of adulticides only when the state declares a public health emergency and there are no practical and effective alternatives to reduce mosquito populations on the Great Meadows Unit of the refuge. • Identify priority areas for enhancement or restoration to reduce the need for mosquito management and improve habitat for native wildlife, fisheries, and plants

1.4 Issues and Concern

1.4.1 Public Participation Public comments and review of this draft will be solicited. Substantial comments and information will be incorporated into the final MMP as appropriate.

1.4.2 Issues Related to the Proposed Action Specific issues associated with mosquito population management on the Refuge include: • Understanding how Refuge-based mosquito populations contribute to or pose a mosquito-borne disease threat to surrounding human developments • Effects of mosquito population monitoring, disease surveillance and control activities on migratory birds, endangered species, and other wildlife and associated habitats • Inter- and intra- agency communication regarding mosquito management activities • Planning and implementation of wetland enhancement and restoration projects that reduce the persistence of above normal mosquito populations • Reliable, consistent management of mosquito program by the CT MMP and the Refuge

1.5 Summary of Laws, Regulations, and Policies Governing the Proposed Action

This section lists the laws, regulations, and policies relevant to mosquito management planning.

1.5.1 National Wildlife Refuge System Administration Act of 1966 The National Wildlife Refuge System Administration Act of 1966, as amended by the National Wildlife Refuge System Improvement Act of 1997 (Act) (16 U.S.C. 668dd-668ee) articulates management priorities for units of the Refuge System and governs refuge uses. Specifically, the Act prohibits uses that are not compatible with the purpose(s) of an individual refuge, maintenance of biological integrity, and the following mission of the Refuge System. “The mission of the System is to administer a national network of lands and waters for the conservation, management, and where appropriate, restoration of the fish, wildlife, and plant resources and their habitats within the United States for the benefit of present and future generations of Americans.” The Service manages the Refuge System for wildlife conservation and gives priority consideration to wildlife-dependent recreational uses in refuge planning and management (see 605 FW 1 -7 in the Service Manual).

1.5.2 National Wildlife Refuge System Regulations Title 50 Code of Federal Regulations (CFR) Subchapter C, 25-38, Administrative Provisions, are Refuge System regulations as authorized by the Administration Act.

• 50 CFR 25.21 (a), (b), and (c) allow a Refuge manager to take actions to protect public health, such as opening or closing a Refuge, or temporarily allowing a particular use, such as mosquito management. • 50 CFR 25.31 establishes requirements for notifying the public about changes to Refuge uses. • 50 CFR 25.41-43 establish responsibility and requirements for issuing or revoking Refuge permits; they also describe the appeals procedure. • 50 CFR 26.41 tells us how to determine whether a Refuge use is a compatible use, meaning it will not materially interfere with or detract from the fulfillment of the mission of the Refuge System or the purposes of the Refuge. • 50 CFR 27.51 prohibits, except by special permit, disturbing, injuring, spearing, poisoning, destroying, collecting or attempting to disturb, injure, spear, poison (such as through the intended use of pesticides), destroy or collect any plant or animal on a Refuge is prohibited.

1.5.3 Endangered Species Act (ESA) of 1973, as amended (16 U.S.C.1531-1544) The ESA provides for the identification, protection, and recovery of species approaching extinction. Protection of federally listed, proposed and candidate species and designated critical habitat can be achieved, in part, through section 7 of the ESA, which requires Federal agencies to consult with the Service or the NOAA Fisheries to ensure that any action an agency authorizes, funds, or carries out is not likely to jeopardize the continued existence of any federally listed listed species, or result in the destruction or adverse modification of designated critical habitat. Agencies consult with the NOAA Fisheries for species that are marine, including anadromous fish, most marine mammals, and sea turtles (when not nesting); otherwise, they consult with the Service.

The actions proposed in an MMP, including but not limited to surveillance, monitoring, and control activities, are subject to a section 7 consultation whenever an activity may affect even a single individual of a listed, proposed, or candidate species or modify designated critical habitat. This consultation helps us to adequately evaluate risk, assess the effects of the physical activities, and also assess ecotoxicological effects of pesticide products to these species and their critical habitats. This level of analysis is necessary for fauna and flora, as effects can be unusual and unexpected.

Even if a public health authority declares a public health emergency, the response is still subject to emergency consultation with the NOAA Fisheries and/or the Service for endangered species issues. Emergency consultations, by regulation, may occur shortly after response. Completing section 7 compliance documentation in conjunction with an MMP may allow the Refuge Manager to avoid emergency consultation if and when there is an unforeseen public health emergency.

1.5.4 Biological Integrity, Diversity, and Environmental Health policy (BIDEH, 601 FW 3) The Service’s BIDEH policy provides for maintenance and restoration of healthy, functioning biological communities composed of native species and habitats. The BIDEH policy favors refuge management which restores or mimics natural ecosystem processes or functions. The BIDEH policy generally discourages controlling native species or using pesticides, yet acknowledges these actions may at times be necessary. Refuge managers must assess any proposed mosquito management actions to ensure they meet the BIDEH policy requirements.

1.5.5 Integrated Pest Management (IPM) policy (569 FW 1) The IPM policy allows the Service to manage pests. It defines pests as any living organism that may interfere with site-specific purposes, operations, or management objectives or that jeopardizes human health and safety. Under 569 FW 1.3 and 1.6, we manage pests when they interfere with site management goals and objectives, jeopardize human health or safety. We also manage pests when their populations exceed action thresholds. An action threshold is the level of damage or number of pests at which we will carry out a management strategy to control the pest population. We allow use of pesticides only after we evaluate a range of alternatives, including physical and cultural methods, biological controls, or taking no action at all. In doing so, we consider human safety, environmental integrity, 12

effectiveness of the action, and cost. The Service’s IPM policy requires a Pesticide Use Proposal (PUP) be approved before a pesticide can be applied on a refuge. Depending on the specific pesticide proposed for use and the application method(s), approval of a PUP may reside with the Refuge Manager, Regional IPM Coordinator, or National IPM Coordinator.

1.5.6 Appropriate Refuge Uses policy (603 FW 1) The Service’s Appropriate Refuge Uses policy provides evaluation procedures (603 FW 1.11A(3)) for refuge managers to ensure that new or existing management actions or methods are appropriate refuge uses. There are five types of refuge uses, and mosquito management to protect public health would be covered under 603 FW 1.10D, Specialized Uses. Because a Refuge Manager must find a use appropriate before undertaking a compatibility review, he/she would need to find mosquito monitoring and management to be appropriate as a precursor to compatibility.

A proposed use must meet at least one condition listed to be determined appropriate; the conditions include being: • A wildlife-dependent recreational use as identified in the Improvement Act, • A use that contributes to fulfilling the refuge purpose(s), • A use that involves the take of fish and wildlife under State regulations, or • A use that an analysis has been found to be appropriate.

The Refuge Manager will address the condition criteria and analysis by completing Service Form 3-2319 for each proposed use under review (see section 1.11 of 603 FW 1). A Refuge Manager retains the authority to reject or modify a use in accordance with this policy.

1.5.7 Compatibility policy (603 FW 2) The Service’s Compatibility policy and associated regulations (50 CFR 26.41) provide guidelines and direct Refuge Managers to ensure that a new or existing activity will not interfere with or detract from the fulfillment of refuge purpose(s) and the mission of the Refuge System. It also requires that we periodically review any use considered compatible to ensure that it complies with all applicable laws, policies, and regulations. If an action is found to be appropriate and compatible, then the Refuge Manager may issue a Special Use Permit (SUP).

Mosquito monitoring and control activities proposed may qualify as a “Refuge use” in accordance with 603 FW 2. Compatibility Determinations (CD) must allow opportunity for public comment and be finalized in writing.

The Compatibility policy also states that a Refuge Manager must determine a use is not compatible if there is insufficient information to determine compatibility. If there are insufficient management resources (e.g., funds, staff, facilities, and equipment) to ensure that a use would occur in a compatible manner, then the use is not compatible. The Compatibility policy states that a use would not be compatible if it conflicts with maintenance of refuge biological integrity, diversity, and environmental health (BIDEH) policy.

If a Refuge Comprehensive Conservation Plan (CCP) included a CD on mosquito management activities, refuge managers should include that documentation in their MMPs. The Refuge does not currently have a completed CCP.

When a public health agency or mosquito control organization proposes to conduct mosquito management activities on a refuge in support of the refuge purpose(s) and in the role of a Service-authorized agent, then that use qualifies as a “Refuge management activity” and the Compatibility policy requirements do not apply. This may be applicable when mosquito monitoring is being conducted at the request of the Service. In this case, the mosquito surveillance, monitoring, and control are being requested by the State.

1.5.8 Special Use Permits The Refuge Manager issues Special Use Permits (SUPs) to authorize special uses on a refuge (see http://www.fws.gov/Refuges/visitors/permits.html (accessed 6/14/2017). The SUPs are issued for three categories of uses: • Commercial Activities, • Research and Monitoring, and 13

• Other General Activity.

A Refuge manager may issue a SUP to allow appropriate and compatible surveillance and monitoring of larval, pupal, and adult mosquitoes and, if necessary, mosquito control activities. To avoid harm to wildlife or habitats, access to traps and sampling stations must meet the compatibility requirements found in 603 FW 2 and the activities are subject to Refuge-specific restrictions.

The instrument we use to document approval of the activity depends on why the activity is taking place. If only monitoring and surveillance are conducted, and no treatment actions will be implemented, monitoring and surveillance activities may be permitted under a Research and Monitoring SUP (FWS Form 3-1383-R, http://www.fws.gov/forms/3-1383-R.pdf, Memorandum of Understanding or other agreement). If a public health agency or mosquito control organization is conducting mosquito management activities on a refuge in support of the refuge purpose(s) and in the role of a Service authorized agent, then an agreement or contract is an appropriate instrument to guide their activities. Others who conduct mosquito management on a refuge for other reasons must get an Other General Activity SUP (FWS Form 3-1383-G, http://www.fws.gov/forms/3-1383-G.pdf.).

1.5.9 National Environmental Policy Act (NEPA) of 1969, as amended (42 U.S.C. 4321-4347) NEPA requires Federal agencies to consider the environmental effects of a proposed action in conjunction with an environmental review addressing among other things, impacts on social, cultural, economic, and natural resources. Agencies must consider a range of reasonable alternatives and the effects of their implementation.

A primary source of information for NEPA is found at https://ceq.doe.gov/ and in the Service’s NEPA for National Wildlife Refuges, A Handbook (http://www.fws.gov/policy/NEPARefugesHandbook.pdf).

Categorical exclusions (CatEx) are categories of Federal actions that do not have a significant effect on the quality of the human environment (individually or cumulatively). As such, they can be excluded from preparation of an Environmental Assessment (EA) or an Environmental Impact Statement (EIS). A CatEx is an environmental review that does not require the extent of analysis that occurs in an EA or an EIS. The Service’s CatEx list is in the Departmental Manual at 516 DM 8.

Although mosquito surveillance or monitoring activities may qualify for consideration as a CatEx, most mosquito surveillance, monitoring and, in particular, treatment actions would have measurable environmental impacts, so they are precluded from CatEx consideration. In this instance, the Refuge has prepared this EA to assess impacts to the environment from the proposed activities.

If the Refuge has completed an MMP that reflects current activities and is NEPA-compliant, then it may be necessary to periodically review the MMP to ensure that it continues to comply with NEPA.

1.6 Decision to be Made

Under NEPA, the Service must decide whether implementing the Proposed Action would have a significant impact to the human environment. If we conclude that the Proposed Action does not have a significant impact to the human environment then we will sign a finding of no significant impact (FONSI) and begin implementation immediately.

Chapter 2 ALTERNATIVES INCLUDING THE PROPOSED ACTION

2.1 The Process Used to Develop the Alternatives

Alternatives were developed to meet the purpose and need, based on U.S. Fish & Wildlife Service laws, regulations, and policies relevant to mosquito management planning (see section 1.5 and Appendix A). A significant amount of information was provided by the CT MMP and included mosquito ecology, history of mosquito populations and their management on the Refuge, cultural tolerances for mosquitoes, past and current historical human health threats, monitoring techniques, treatment thresholds and disease surveillance.

2.2 Description of Alternatives, Including the Proposed Action and No Action

2.2.1 Factors Common to All Alternatives Actions that are common to all alternatives are described below and are not repeated in each alternative description.

2.2.1a General Permits. The CT MMP must obtain all permits required for state and federal endangered species compliance before allowing mosquito management activities in endangered species habitat on the Refuge. Other general permits may also be required such as a National Pollutant Discharge Elimination System (NPDES) permit, depending on the scope of the action proposed each year.

2.2.1b Special Use Permits. The CT MMP must obtain an annual Refuge Special Use Permit (SUP; see Appendix E) if they will be conducting mosquito management activities (which include surveillance, monitoring, and control) on the Refuge. A SUP will be issued, renewed, and/or revised annually and will document all uses on the Refuge and provide clear guidance for activities on the Refuge. To ensure that mosquito management activities are compatible with the Refuge purposes, permitted activities must meet the stipulations listed in the Compatibility Determination. For more details on SUP, see Annual Meeting on page 31 of this document.

2.2.1c Supplemental NEPA Documentation. Each of the alternatives described below may require a supplemental NEPA depending on the scope of the action proposed each year.

2.2.1d Education and Outreach. Where appropriate, we will collaborate with Federal, State, and/or local wildlife agencies, public health authorities, agriculture departments, and vector control agencies to conduct education and outreach activities aimed at protecting human health from threats associated with mosquitoes. Where appropriate, we will provide access to information materials about mosquito-associated threats to our visitors and employees (e.g., Refuge office, internet sites, and signage). The Refuge will prepare an instructional package for employees on personal protection measures to minimize their exposure to mosquito- borne diseases.

2.3 Alternative A – Proposed Action – Phased Approach to Mosquito Control

The Proposed Action is to implement a mosquito management plan (Plan) at Great Meadows Unit of the Refuge that consists of a phased approach to mosquito management and is consistent with the principles of IPM. The Proposed Action emphasizes design, restoration, and management of wetlands in a manner to benefit wildlife and minimize mosquito production. Wetland restoration or enhancement projects would be implemented as funding becomes available. The improvement of hydrology within wetlands would be the primary mechanism for enhancing habitat and reducing mosquito production. Restoration projects would be focused on improving habitat for native wildlife and plants as well as minimizing mosquito production. The Refuge will focus restoration activities for areas of the site that are current mosquito management areas (for map of these areas, see Appendix C; p. 38).

This alternative is consistent with an IPM approach, which is a sustainable approach to managing pests by combining biological, cultural, physical, and chemical tools in a way that minimizes economic, health, and 15

environmental risks. When practical, the approach may include compatible actions that reduce mosquito production, which minimizes or eliminates the need to apply pesticides. We consider the procedures described below as long- term practices to reduce persistent potential mosquito-associated health threats that Federal, State, and/or local public health authorities have identified.

While the emphasis of this preferred alternative is restoring wetlands, the phased approach also includes monitoring, surveillance, and the warranted application of pesticides. The application of pesticides would be approved based on the phased approach outlined below. The principle goal of a phased approach to mosquito management is to minimize effects on Refuge resources while addressing legitimate human health concerns and complying with Service regulations and policies. The implementation of a phased-response program represents a standardized approach that would result in a consistent mosquito management program that adheres to Service and Connecticut state guidelines. Because occurrence of arboviruses and other human health issues resulting from mosquitoes are sporadic, phases of mosquito management implemented on the Refuge could vary from year to year.

The following phased mosquito management program is dependent upon communication and cooperation with public health agencies and CT MMP (which includes the CT Department of Health). The Refuge would actively engage in the implementation of a mosquito management program with CT MMP. Although the CT MMP would have the lead for monitoring, disease surveillance, and pesticide applications, evaluation of monitoring data and approval for each management action would be the responsibility of the Refuge Manager. Although additional staff time would be required to oversee the mosquito management program, due diligence is necessary to ensure that the conditions for compatibility are met and the program is implemented so as to avoid or minimize effects on Refuge resources.

Table 1 below provides abbreviated descriptions and responses associated with each of the mosquito management phases. Because of the nature of mosquito-borne diseases, as well as the limited information available regarding the effects of these diseases on wildlife of the Refuge, this approach focuses on the implementation of a mosquito management program to protect the public from mosquito-borne disease.

Mosquito monitoring represents the baseline activity of mosquito management on the Refuge and would occur prior to and along with the following management phases. Monitoring is required to determine mosquito population estimates and locations of infestations. Because the CT MMP would have the lead for monitoring mosquito populations, communication and cooperation are imperative to develop reliable information to determine appropriate management level(s). All mosquito management decisions would be made in consultation with the Refuge and appropriate health and vector control agencies using monitoring data collected on and within the vicinity of the Refuge.

The foundation for the following phased mosquito management approach is a series of IPM options that are intended to minimize effects of mosquito management to Refuge resources while protecting human health. We will use human, wildlife, or domestic animal mosquito associated health threat determinations, combined with refuge mosquito population estimates, to determine the appropriate refuge mosquito management response. We will allow pesticide treatment to control mosquitoes on refuge lands only after evaluating all other reasonable IPM actions. The decision to use pesticide treatments will be based on monitoring data for the relevant mosquito life stage and used only when necessary to protect the health of human, wildlife, or domestic animals.

Table 1. Phased-response mosquito management guidelines for the Great Meadows Unit at

thePhase Stewart B. McKinney ConditionNWR Response

1 No documented existing health threat1. Monitoring and surveillance of areas surrounding

Mosquito management issues have not been the Refuge to inform management actions on the

reported or identified by the CT MMP. Refuge. Remove/manage artificial breeding sites

such as tires, tanks, or similar debris/containers.

Consult with CT MMP when planning wetland

enhancement or restoration projects. See State

WNV and EEE plans in Appendix F for more

details.

2 Potential human or wildlife (incl. threatened and Response as in threat level 1, plus: allow endangered species) health threat1 (presence of compatible monitoring and disease surveillance. vector spp, historical health threat, etc.), as Consider compatible non-pesticide management documented by the CT MMP. options to reduce the potential for above-normal mosquito production (e.g., restore/enhance tidal marsh hydrology). See State WNV and EEE plans in Appendix F for more details. 3 Mosquito larvae threshold2 exceeded for human Response as in threat level 2, plus: allow health on the Refuge as determined by compatible site-specific application of larvicide in standardized monitoring. Documented potential areas with above average mosquito populations, as human or wildlife/domestic animal health threat determined by monitoring. Conduct post larvicide (historic health threat, presence of vector species). monitoring to determine efficacy. See State WNV and EEE plans in Appendix F for more details.

4 Mosquito larvae have begun to reach last instar Response as in threat level 3, plus: if appropriate, stages or pupate reducing the efficacy of increase the intensity and frequency of larvicides, larvicides. Mosquito larval and pupal population allow compatible site-specific use of pupacides in thresholds2 exceeded on the Refuge. Mosquitoes areas with above average mosquito populations, 1 produced by the Refuge pose a health threat as determined through monitoring to be beyond determined by the CTMMP. Before the use of control with larvicides. Increase monitoring and pupacides, the Directors of the CT DEEP and CT disease surveillance. Conduct post larvicide and DPH will approve the use in coordination with pupacide monitoring to determine efficacy. See the Refuge Manager. This approval will be State WNV and EEE plans in Appendix F for more documented by the CT DEEP. details. 5 Exceedance of larval, pupal, and adult mosquito Response as in threat level 4, plus: Consider site- 2 specific adulticiding in areas (with a pyrethrin- population thresholds on the Refuge. High risk based adulticide) in areas with above average for mosquito-borne disease (imminent risk of serious human disease or death, or an imminent mosquito populations as determined by risk of serious disease or death to populations of monitoring. Conduct post adulticide monitoring wildlife) within communities surrounding the to determine efficacy. See State WNV and EEE plans in Appendix F for more details. Refuge has been documented by the CT MMP and the Governor of the State of Connecticut has declared a public health emergency.

1An adverse impact to the health of human or wildlife populations from mosquito-borne disease identified and documented by Federal, State, and/or local public health authorities. Health threats are locally derived and are based on the presence of endemic or enzootic mosquito-borne diseases, including the historical incidence of disease, and the presence and abundance of vector mosquitoes. Health threat levels are based on current monitoring of vectors and mosquito-borne pathogens. All State plans in Appendix F.

2Human health threshold (e.g., numbers per dip) is determined by considering several factors as determined by Appendix F. Larval thresholds are presented in state plans in Appendix F.

The first two phases require use of indirect approaches (e.g., source reduction, adjustments in habitat management programs) to reduce mosquitoes with little or no effect to Refuge resources. The last three phases allow the use of certain pesticides (e.g., larvicides) to address threats to human health. Phase 5 allows for the use of adulticides when: 1) mosquito-borne disease activity is documented on the Refuge or within flight range of vector mosquito species present on the Refuge; 2) mosquito monitoring on the Refuge indicates adult vector populations have exceeded the thresholds identified in the State’s surveillance and response plans Appendix F; 3) there are no practical and effective alternatives to reduce the health threat; and 4) the National IPM Coordinator has approved the use of the adulticide. To avoid the use of adulticides, the approach favors an early response with larvicides (See Appendix F for surveillance and response plans and the CT MMP’s PDF about larvicides here: http://www.ct.gov/mosquito/lib/mosquito/publications/2001larv.pdf). Early response with larvicides would be directed at specific locations where mosquito larvae are present. The delayed use of larvicides may result in the need for adulticides that pose the greatest potential threat to Refuge resources (see Appendix C for mosquitoes found at testing stations near Refuge). Pesticide treatment may not be used on the Great Meadows Unit at Stewart B. McKinney NWR solely for nuisance mosquito relief, but may be considered when there is a demonstrated human health risk, mosquitoes are detrimental to refuge goals and objectives, and mosquito management actions will not interfere with refuge goals and objectives.

2.3.1 Phase 1 In Phase 1, a health threat has not been identified and mosquito management issues have not been reported or identified by the appropriate public health authority or to CT MMP. To avoid any possible mosquito management issues, artificial mosquito breeding habitat throughout the Great Meadows Unit at the Refuge, such as tires, open containers, and other equipment or objects that pool water where mosquitoes may breed, should be eliminated. Although artificial mosquito breeding habitat is not currently an issue on the Refuge, this should still be a concern into the future as a preventative measure.

The Refuge would consult with the CT MMP when wetland enhancement or restoration projects are being planned on the Refuge. Consultation would allow Refuge staff and the CT MMP to identify potential issues or opportunities related to mosquito production and management in the future. Monitoring and surveillance of mosquito abundance and disease prevalence in areas similar and near the Refuge would be conducted by the CT MMP which would inform the potential for mosquito management needs on the Refuge.

2.3.2 Phase 2 In Phase 2, the Refuge Manager is contacted by the appropriate public health authority(ies) or CT MMP regarding a potential human health threat posed by mosquitoes harbored or produced on the Refuge. In response, Refuge staff may increase compatible mosquito population monitoring and disease surveillance by the CT MMP. The initial step to developing a proactive prevention and management program for mosquitoes is to determine mosquito species presence and abundance at the Great Meadows Unit on Refuge lands, and to identify potential or documented vectors of mosquito-borne diseases that represent a potential human health threat. This would be accomplished at the State monitoring locations – of which there are two – adjacent to the Great Meadows Unit. These stations will determine adult species type, abundance, and if the mosquiotes harbor disease vectors. Monitoring and surveillance activities should be well-documented and presented to Refuge staff by the CT MMP. This data will help determine the need to control larval mosquitoes.

In order to avoid or minimize the use of pesticides, habitat management practices or wetland enhancement/restoration projects that improve wildlife habitat and reduce seasonal abundance of larval and adult mosquitoes should be implemented where possible.

Refuge staff and visitors would be informed of an increased health threat associated with mosquito-borne disease activity. Personal protection measures such as wearing mosquito repellant would be recommended to staff and visitors.

2.3.3 Phase 3 If non-pesticide attempts to reduce mosquito populations are unsuccessful or are not feasible and mosquito larvae thresholds have been exceeded (Appendix F), application of larvicides would be considered. The treatment of marshes on the Great Meadows Unit of the refuge using larvicides would only be permitted after the CT MMP has provided the refuge manager with information as described in the Connecticut West Nile Virus Surveillance and response Plan (CT DEEP 2012) that shows mosquito larvae populations are widespread within the salt marsh (areas C and D), and after monitoring data reveals that mosquito populations meet or exceed the following abundance measures: using a long-handle, plastic, 8 ounce cup dipper - if 5 or more larvae/dip are found in 50 percent or more of the dips (minimum of 3 dips/site). Other factors to be considered in making the determination to allow larvacide treatment include marsh hydrology (drying versus flooding), rainfall, temperature, instar larval stages, and treatment history. Locations of larvicide treatments would be based on standardized monitoring results (see Appendix F). The preferred larvicide treatments are biorationals (biological agents) Bti and Bs because of limited non-target effects (Appendix H, I). Post larvicide monitoring would be conducted to determine efficacy.

2.3.4 Phase 4 If appropriate, the intensity and frequency of larvicides would be increased. Larvicides (Bti or Bs) are most effective on mosquitoes during early instar stages (up to the fourth) and do not control pupae. If developing mosquitoes have reached late instar stages or have pupated, then we would consider using pupacides in areas with above average mosquito populations (determined through monitoring). Because pupacides can negatively impact all invertebrates that require surface air (e.g., act as surfactants), the use of these pesticides should be carefully considered. Due to the broad-spectrum action of surface oils and films, they typically are not appropriate and are rarely authorized for use on refuges. For this reason, pupacides (Agnique) would only be used if large numbers of mosquitoes are considered an immediate threat to human health and thresholds developed by the appropriate public agency have been exceeded (there is active transmission of mosquito-borne disease from the Great Meadows Unit at the Refuge based mosquitoes or within flight range of vector mosquito species present on the Refuge). Post larvicide and pupacide monitoring would be conducted to determine efficacy and any adverse impacts. Before the use of pupacides the Directors of the DEEP and DPH will approve the use in coordination with the Refuge and that approval will be documented by DEEP.

2.3.5 Phase 5 In this phase, mosquito-borne disease activity has been documented on the Great Meadows Unit at the Refuge or within flight range of vector mosquito species present on the Refuge. There is active transmission of mosquito borne disease on the Great Meadows Unit at the refuge from refuge based mosquitoes. A risk of serious mosquito-borne human disease or death has been documented by the appropriate public health authority. Disease surveillance determines that there is a high risk for mosquito-borne disease within the vicinity of the Great Meadows Unit at the Refuge. For example, pathogen presence in mosquito pool(s), wild birds, horses, or humans has been documented within the flight range of vector mosquito species present on the Great Meadows Unit at the Refuge. These conditions in combination with adult mosquito populations above threshold levels determined by the CT MMP on the Great Meadows Unit at the Refuge would trigger consideration of a more aggressive treatment strategy, including the use of adulticides. If larvicide and/or pupacide treatments fail, pyrethrin/pyrethroid adulticides would be considered for use on the Great Meadows Unit at the Refuge to suppress populations of adult mosquitoes and interrupt epidemic virus transmission. Because the efficacy and effects of adulticides are variable, adulticides should not be applied broadly without site-specific data indicating a need for control. Further, the use of adulticide would be considered in concert with the West Nile Virus Surveillance & Response Plan, Contingency Plan for Eastern Equine Encephalitis, and Zika Virus Surveillance and Response Plan and only after the Governor has issued a public health emergency (Appendix F). The CT MMP would be required to prepare a Risk Assessment as part of their request to apply adulticides. The Risk Assessment evaluates a number of factors including environmental conditions, species presence, virus infection rate, dead bird presence, and human cases to determine whether adulticide should be considered. We would only consider application in areas where a pathogen is present and mosquito population thresholds have been exceeded on the Great Meadows Unit at the Refuge that can be effectively treated while minimizing non-target effects, especially to threatened and endangered species. However, specific areas treated and

the extent of treatment would vary from year to year depending on mosquito populations and environmental conditions.

In order to limit human contact with adulticides, visitors would not be allowed in those parts of the Great Meadows Unit at the Refuge that are being treated with adulticides. Information about treatment scheduling, location, and pesticide would be posted on the Refuge website, at the Refuge Headquarters, and at the treatment location. Post adulticide monitoring would be conducted to determine efficacy and any adverse impacts.

In summary, application of adulticides on the Refuge would require the following steps:

• Prior approval from the National IPM Coordinator via an approved Pesticide Use Proposal • The CT MMP must present to the Refuge Manager with data supporting presence of an arboviral disease on the Refuge or within flight range of the vector mosquito species on the Refuge, including a Risk Assessment in the region • The CT MMP must provide the Refuge Manager with types/quantities and locations of adulticides proposed • If beneficial, the CT MMP should conduct simultaneous application of larvicides with the adulticide application to prevent future adult outbreaks

2.3.6 Access Access for the purposes of mosquito management (e.g., monitoring, surveillance, control) would be limited in areas known to support the State listed plant species, salt marsh pink (Sabatia stellaris) or in known saltmarsh sparrow (Ammodramus caudacutus) and seaside sparrow nesting areas (during the nesting season) and/or in close proximity to sloughs and channels to avoid trampling and effects on non-targeted species.

The following access limitations apply: • Motorized access only in areas designed for motor vehicles e.g. railroad trail and other access roads. • Unless permitted by the Refuge Manager, mosquito management activities should not occur within 100 feet of natural sloughs and channels. • All personnel entering the wetlands would be trained by Refuge staff and CT DEEP staff at the annual meeting to avoid disturbance to endangered, threatened or other sensitive species of the Refuge. • Application is only to be conducted as approved in SUP/PUPs

These access limitations would limit direct and indirect (e.g., habitat) negative effects on sensitive species. Access within sensitive areas would be identified by the Refuge Manager in coordination with the CT MMP and designated in the annual Special Use Permit.

2.3.7 Mosquito Monitoring and Surveillance We would allow compatible monitoring and surveillance of larval and adult mosquito populations on the Great Meadows Unit at the Refuge under a Refuge Special Use permit. To avoid harm to wildlife or habitats, access to traps and sampling stations must meet the requirements outlined in the Compatibility Determination (appendix C) and may be subject to Refuge-specific restrictions.

Mosquito population monitoring involves activities associated with collecting quantitative data to determine mosquito species composition and to estimate relative changes in mosquito populations over time. Mosquito population monitoring on the Great Meadows Unit of the refuge will document mosquito species composition to genus or species level, and estimate population size and distribution across refuge wetland habitats during the breeding season, using standard methods employed by mosquito control professionals. The objectives of mosquito population monitoring are to:

1. Establish baseline data on species and abundance, 2. Map breeding and/or harboring habitats, 20

3. Estimate relative changes in population sizes and associated health risks for making IPM decisions to reduce mosquito populations when necessary, 4. Determine effectiveness of treatment, and 5. Use this information to guide integrated pest management of mosquito populations.

The purpose of mosquito-borne disease surveillance involves activities associated with detecting pathogens causing mosquito-borne diseases, such as testing adult mosquitoes for pathogens or testing reservoir hosts for pathogens or antibodies. These activities assist in determining public health risks associated with mosquito-borne pathogens on or near the Refuge.

Monitoring of immature mosquitoes on the Great Meadows Unit at the Refuge would be conducted by the CT MMP (typically from April through October). Field technicians within these agencies maintain a list of known mosquito developmental sites on the Refuge and visit them during predominant periods of mosquito production. The timing and frequency of monitoring is based on a number of factors including history of mosquito production, tidal cycles, precipitation levels, and available monitoring resources. In Connecticut, mosquito larvae/pupae need at least 5 to 7 days in water to fully develop; they cannot survive in situations where surface water drains off or evaporates within that time frame. Therefore, inspections will focus on identifying and sampling shallow depressions, occluded ditches, or similar sites in tidal and non tidal habitat that could hold tidal or rain water for more than 5 to 7 days. Inspections will occur during normal working hours. Mosquito populations are sampled using established protocols (Appendix G). If data reveals that mosquito populations meet or exceed the following abundance measures: using a long-handle, plastic, 8 ounce cup dipper - if 5 or more larvae/dip are found in 50 percent or more of the dips (minimum of 3 dips/site). Other factors to be considered in making the determination to allow larvacide treatment include marsh hydrology (drying versus flooding), rainfall, temperature, instar larval stages, and treatment history.Samples are examined in the laboratory by the CT MMP to determine the abundance, species, and the presence of viruses. Field examinations are made for population and life cycles. This information is compared to historical records and established thresholds (Appendix F) and would be used as a tool for treatment decisions.

Although larval mosquito control is preferred, identifying all larval sources in a timely manner is not possible. Therefore, adult mosquito monitoring is also needed to pinpoint problem areas and locate previously unrecognized or new larval developmental sites. These trap sites are located within a mile radius of the Refuge. Adult mosquitoes are sampled using standardized trapping techniques (i.e., carbon dioxide-baited light traps and Gravid traps; Appendix I). Mosquitoes collected using these methods are counted and identified to species. Information on adult mosquito abundance from traps is augmented by tracking mosquito complaints from local residents. Analysis of requests for mosquito control allows CT MMP staff to gauge the success of control efforts and locate undetected sources of mosquito development. CT MMP conducts public outreach programs and encourages local residents to contact them to request services.

Monitoring will be conducted by the CT MMP, primarily on foot. Vehicle access to the marsh edge will be restricted to established roads and fire trails at the Great Meadows Unit at the refuge. A more detailed presentation of monitoring and disease surveillance protocols used by the CT MMP is presented in Appendices F and G.

2.3.8 Wildlife Monitoring and Surveillance Wildlife monitoring will be conducted to assess any potential impacts from mosquito management activities. The Refuge and the CT MMP will monitor for impacts to state listed species, particularly salt marsh and seaside sparrow and salt marsh pink, due to mosquito control (including mosquito monitoring and surveillance). All wildlife surveys will follow established protocols. Monitoring plans will be developed as a result of the annual meeting in conjunction with the CT MMP.

2.3.9 Pesticide Approval Process As a result of its statute authority under the Migratory Bird Treaty Act, the Endangered Species Act and Service policy, the Service is required to consider whether use of specific pesticides would harm trust species. The Service

evaluates approval of specific pesticide use based on its history of adverse effects on non-target species and persistence in the environment.

Refuge staff would prepare Pesticide Use Proposals (PUPs) on an annual basis (in coordination with the CT MMP) for Service approval. The PUPs would include pesticides that CT MMP or other permitted groups propose for use as part of a mosquito management program for the Great Meadows Unit at the Refuge. Pesticide Use Reports (PURs) would be prepared by Refuge staff in coordination with the CT MMP on an annual basis following application of pesticides to control mosquitoes on the Refuge. To assist in tracking mosquito management activities, the CT MMP would prepare an annual quantitative summary of Refuge mosquito monitoring and surveillance results, control activities on the Great Meadows Unit at the Refuge (e.g., pesticides applied, amount of pesticides applied, locations of application, method of application), and regional disease surveillance. The report should be accompanied by maps showing specific areas where management activities occurred. All surveillance and control activities would be spatially referenced (e.g., use of GPS, GIS). Comparisons of mosquito management within and among years should be presented to permit analysis of patterns that may indicate success of habitat management efforts or suggest the need for a new management approach.

Methods used to reduce mosquito populations are primarily based on efficacy, cost, and minimal ecological disruption, including minimum effects on non-target organisms and natural systems of the Refuge. Chemical pesticides should be used only where practical physical, cultural, and biological alternatives or combinations thereof, are impractical or incapable of providing adequate mosquito population control. Furthermore, chemical pesticides would be used primarily to supplement, rather than as a substitute for, practical control measures of other types. Whenever a chemical is needed, the most narrow ranging and specific pesticide available for the target organism in question should be chosen, unless consideration of persistence or other hazards would preclude that choice." (7 RM 14.2).

2.3.10 Mosquito Control Pesticides Mosquito control pesticides can be categorized into 3 groups: larvicides, pupacides (surface films/surfactants), and adulticides. Compared with other forms of pest control, there are relatively few pesticides available within each of these categories, and all differ with regard to efficacy and effects on non-target organisms. Additional information on pesticides presented here can be found in Appendices H, I, and K. Pesticides commonly used by local CT MMP are presented in Table 2.

Table 2. Common pesticides used for mosquito control in wetlands of Connecticut.

a Name Trade Name Formulation Application Bacillus thuringiensis Vectobac CG Granules Larvae israelensis (Bti)

Bacillus sphaericus VectoLex FG Fine granules Larvae (Bs)

Monomolecular film Agnique Liquid Larvae, pupae

Pyrethroids ANVIL 2+2 ULV; Scourge; ULV, ULV, LC, LC Adults Masterline Bifenthrin 7.9; ExciteR™

Table adapted from Rose (2001) (http://www.cdc.gov/ncidod/eid/vol7no1/rose.htm) aAS = Aqueous suspension; B = Briquets; EC = Emulsifiable concentrate; G = Granules; LC = Liquid concentrate; P = Pellets; ULV = Ultra low volume; WDG = Water-dispersible granule

The use of larvicides would be routinely approved subject to review by the Regional Office Integrated Pest Management Coordinator acting under the Service’s Washington Office. Data from various sources (e.g., scientific literature) would be used to identify whether new preferred chemicals exist, as they become available. New control products will be considered based on their effects compared to those products identified in the plan.

Before applying pesticides to the Great Meadows Unit at the Refuge in a non-emergency situation Refuge staff must:

1. Use current monitoring data for larval, pupal, and adult mosquitoes which documents the need for mosquito management. 2. Determine the most appropriate pesticide treatment options based on monitoring data for the relevant mosquito life stage. This will be accomplished through the Pesticide Use Proposal preparation system. Only pesticides described in the preferred alternative will be utilized for the control of larval and pupal stages. The control of adult mosquitoes would only be considered if a public health emergency were declared. At that time, a Pesticide Use Proposal would be developed for adulticides. 3. Consider whether use of pesticide would harm trust species. 4. Have an approved pesticide use proposal (PUP) in place.

2.3.10a Larvicides. Larvicides are materials that affect the four larval stages of mosquitoes known as instars. They can be applied through a wide variety of methods including hand application and backpack sprayers, amphibious tracked vehicle, and truck-mounted equipment. Mosquito larvicides relevant to this EA include Bti (Bacillus thuringiensis var. israelensis), and Bs (Bacillus sphaericus). The treatment of marshes on the Great Meadows Unit of the refuge using larvicides would only be permitted after the CT MMP has provided the refuge manager with information as described in the Connecticut West Nile Virus Surveillance and response Plan (CT DEEP 2012) that shows mosquito larvae populations are widespread within the salt marsh (areas C and D), and after monitoring data reveals that mosquito populations meet or exceed the following abundance measures: using a long-handle, plastic, 8 ounce cup dipper - if 5 or more larvae/dip are found in 50 percent or more of the dips (minimum of 3 dips/site). Other factors to be considered in making the determination to allow larvacide treatment include marsh hydrology (drying versus flooding), rainfall, temperature, instar larval stages, and treatment history. Larvicides may be approved through a PUP by the Refuge Manager of the Stewart B. McKinney NWR. Bti and Bs will be applied on the Great Meadows Unit at the Refuge. We favor using the larvicide that would have the least adverse impacts on non target invertebrates, produce fewer disruptions to food webs critical for migratory birds, and reduce lethal effects on natural mosquito predators, such as larval forms of odonates, hemipterans, and coleopterans. CT MMP will conduct post larvicide monitoring to determine effectiveness. Refer to Appendix H for a more detailed account of non-target effects of larvicides used for mosquito control.

Bacillus thuringiensis (Bt) is a natural soil bacterium that acts as a larval stomach poison. Bt must be ingested by the larval form of the insect in order to be effective. Bt contains crystalline structures containing protein endotoxins that are activated in the alkaline conditions of an insect’s gut. These toxins attach to specific receptor sites on the gut wall and, when activated, destroy the lining of the gut and eventually kill the insect. The toxicity of Bt to an insect is directly related to the specificity of the toxin and the receptor sites. Without the proper receptor sites, the Bt will simply pass harmlessly through the insect’s gut. Several varieties of Bt have been discovered and identified by the specificity of the endotoxins to certain insect orders. Bacillus thuringiensis var. kurstaki, for example, contains toxins that are specific to Lepidoptera (butterflies and moths), while Bti (variety is specific only to certain primitive dipterans (flies), particularly mosquitoes, black flies, and some chironomid midges (Boisvert and Boisvert 2000). Bti is the form that would be used on the Great Meadows Unit at the Refuge. Bti is not known to be directly toxic to non-dipteran insects. Bti is an U.S. Environmental Protection Agency (USEPA) toxicity class III general use pesticide, does not contain toxic inert ingredients, and is practically non toxic to animals (Extoxnet 1996).

Because Bti must be ingested to kill mosquitoes, the treatment is much more effective on first-, second-, and early third-instar larvae than on late third and fourth instars since the earlier instars feed at a faster rate (fourth instar larvae feed very little). The pesticide is completely ineffective on pupae because they do not feed at all.

Formulated products may be granular or liquid, and potency is expressed in International Toxicity Units (ITU), usually ranging from 200-1200 ITU. The concentrations of Bti in water necessary to kill mosquito larvae vary with environmental conditions, but are generally 0.05-0.10 ppm. Higher concentrations (0.1->0.5 ppm) of Bti are necessary when there is a high amount of organic material in the water, late-third and early fourth instar larvae predominate, larval mosquito density is high, or water temperature is low (Nayar et al. 1999). Operationally, Bti is applied within a range of volume or weight of formulated product per acre as recommended on the pesticide label, with the goal to achieve an effective concentration. The label recommended range of application rates under most conditions varies by a factor of 4 for most formulations (e.g., for granular formulations, 2.72-11.12 kg/ha (2.5-10 lb/acre)). For later instar larvae and water with a high organic content, higher application rates are recommended that may reach 8 times the lowest rate (e.g., for granular formulations, the higher rate is 11.1-22.5 kg/ha (10-20 lb/acre)). Mosquito control agencies use the recommended label rates, along with previous experience, to administer an effective dose. Because water depths even within a single wetland can vary greatly, field concentrations of Bti can vary widely, especially when the pesticide is applied aerially. Efficacy is monitored by post-application reductions in mosquito larval density, but the actual concentration of Bti following an application is not measured. Thus, an insufficient concentration of Bti can be detected by low mortality of mosquito larvae, but an overdose (i.e., a concentration greater than necessary to kill mosquito larvae) of the pesticide is rarely monitored for.

Bacillus sphaericus is a naturally occurring bacterium that is found throughout the world. Bacillus sphaericus was initially registered by USEPA in 1991 for use against various kinds of mosquito larvae. Mosquito larvae ingest the bacteria, and as with Bti, the toxin disrupts the gut in the mosquito by binding to receptor cells present in insects, but not in mammals. VectoLex CG and WDG are registered B. sphaericus products, and are effective for approximately one to four weeks after application. Because Bs is a more recently developed larvicide than Bti, there are fewer studies that have examined the non-target effects of this pesticide. The data available, however, indicate a high degree of specificity of Bs for mosquitoes, with no demonstrated toxicity to chironomid larvae at any mosquito control application rate (Mulla et al. 1984; Ali and Nayar 1986; Lacey and Mulla 1990).

2.3.10b Pupacides (Surface Films). Surface films are applied to mosquito breeding sites to kill mosquito larvae and pupae. The products create a barrier to the air-water interface and suffocate insects, which require at least periodic contact with the water surface in order to obtain oxygen. Surface films are alcohol based and produce a monomolecular film over the water surface.

Films are potentially lethal to any aquatic insect that lives on the water surface or requires periodic contact with the air-water interface to obtain oxygen.

Monomolecular Film (MMF) is a non-ionic surfactant that has an alcohol base. The film produced by MMF reduces the surface tension of the water making mosquito larvae and pupae unable to attach, thus causing them to drown. Emerging adult mosquitoes or midges are unable to fully emerge and will drown. The film produced by Agnique is not visible on the water surface and should not be used in areas that are subject to unidirectional winds greater than 10 mph or where surface water overflow or runoff is an issue. See Appendix H for non-target effects of Agnique.

2.3.10c Adulticides. Adulticides are pesticides used to kill adult mosquitoes. Adulticides appear to effectively control adult mosquito populations and the spread of mosquito borne diseases such as WNV (Carney et al. 2008), but only for a brief time, and are therefore only recommended during a disease event to break the disease transmission cycle (http://www.townofsilvercity.org/vector/ scienceofvector.html). All pesticides used to kill adult mosquitoes are broad-spectrum insecticides. The only selective aspect of these pesticides is in the manner in which they are applied. Most adulticides under use in Connecticut are applied as ultra-low volume (ULV) sprays and they are sprayed as very fine droplets. Small droplet size allows the spray to drift for a relatively longer period of time compared to larger droplets, and the small size delivers an appropriate dose of the pesticide to kill an adult mosquito. Drift is a necessary component of adulticiding because these sprays are most effective on flying insects. For this reason, adulticide applications generally would occur in the evening or early morning hours when the majority of mosquito species are most active. Adulticides would be primarily applied by truck-mounted or backpack sprayers on the Great Meadows Unit at the Refuge. Aerial spraying of adulticides may occur under emergency situations. Although the local mosquito population is reduced for a few days, these treatment methods do not prevent mosquitoes from re entering the sprayed area.

There are three general classes of adulticides: organophosphates, pyrethroids and pyrethins. These pesticides work on the nervous system although they have different modes of action. Only pyrethrin based adulticides are being considered for use on the Great Meadows Unit at the Refuge at this time. Organophosphates are cholinesterase inhibitors while pyrethroids and pyrethrins are sodium channel blockers. Organophosphates are not used by the CT MMP for mosquito control and they are not permitted for use on the Refuge. Pyrethrins are naturally occurring compounds extracted from chrysanthemum plants and have been used to make pesticides (McLaughlin 1973, Klassen et al. 1996, Todd et al. 2003). Pyrethroids are synthetic products that have the same basic chemical make-up as pyrethrins but are not naturally occurring. Pyrethrum is the general term covering pyrethrins and pyrethroids.

The most common pyrethroids are the synthetic pyrethroids, permethrin, resmethrin, and sumethrin. Both pyrethroids and pyrethrins are usually combined with the synergist piperonyl butoxide, which interferes with an insect's detoxifying mechanisms (Tomlin 1994). Nontarget toxicity from pyrethroids may occur in either terrestrial or aquatic habitats as a result of deposition, runoff, inhalation, or ingestion (Appendix I, K). In general, pyrethroids have lower toxicity to terrestrial vertebrates than the organophosphates. Pyrethroids, although less toxic to birds and mammals, are toxic to fish and aquatic invertebrates (Anderson 1989, Siegfried 1993, Tomlin 1994, Milam et al. 2000). The actual toxicity of pyrethroids in aquatic habitats, however, is less than may be anticipated because of the propensity of these pesticides to adsorb to organic particles in the water (Hill et al. 1994). There are also data that indicate synthetic pyrethroid degradates have endocrine disrupting properties (Tyler et al. 2000).

The natural pyrethrins are non-systemic contact poisons which quickly penetrate the nerve system of the insect and cause paralysis and subsequent death (Extoxnet 1994, and Tomlin 1994). A few minutes after application, the insect cannot move or fly away, but a "knockdown dose" does not mean a killing dose. The natural pyrethrins are swiftly detoxified by enzymes in the insect. Thus, some pests will recover. To delay the enzyme action so a lethal dose is assured, commercial products are formulated with synergists, e.g. piperonyl butoxide, which inhibit detoxification (Tomlin, 1994). Pyrethrins are generally considered less toxic to invertebrates and are less persistent in the environment relative to synthetic pyrethroids although data on toxicity are lacking (Spurlock 2006). Studies of pyrethrin have shown low toxicity to birds and mammals but higher levels of toxicity among aquatic species such as fish and invertebrates (e.g., Gunasekara 2005).

2.3.11 Annual Meeting/Training All alternatives require that an annual meeting be held to discuss mosquito activities for the past year and any proposed wetland and mosquito management changes or issues for the upcoming season. Annual meetings will also allow for any changes that may need to be adapted as a result of changing environmental conditions or new 25

treatment methods and pesticides. Significant changes will require a new or revised mosquito management plan for the refuge. The following is a list of topics that would be covered:

Service: • Staff introduction/changes • Pest management policy review and changes • Summary of current wetland restoration and management program • Proposed enhancement or restoration projects • Current wildlife populations & status • Techniques to minimize disturbance to wildlife

CT MMP: • Summary of mosquito production areas • Summary of mosquito management activities • Staff introduction/changes • Mosquito policy changes • Updated Pesticide Use Proposals and labels • Proposed changes to mosquito management program • Current mosquito and disease information • Listed species monitoring and surveillance Results of relevant mosquito research projects • Proposed mosquito reduction projects • Current mosquito production areas • By this meeting a notice of intent would be submitted to the Environmental Protection Agency for the use of pesticides in the salt marsh in accordance with the Clean Water Act.

2.4 Alternative B – Monitoring and Larvicide Only

Under this alternative, the CT MMP would continue to operate in a manner similar to the years 1998-2007 in which mosquito monitoring and disease surveillance is followed by control with larvicides (Bti) when action thresholds have been exceeded. The CT MMP would coordinate with the Refuge Manager prior to surveillance, control, and monitoring activities on the Great Meadows Unit at the Refuge. This alternative is a phased approach similar to the Proposed Action with the exception that adulticides and pupicides are not permitted on the Refuge. This alternative would include wetland restoration.

2.5 Alternative C – No Action

Under this alternative, only mosquito population monitoring and disease surveillance would be allowed. However, mosquito control using biological or chemical methods would not be permitted on the Great Meadows Unit at the Refuge. Efforts to enhance and restore wetlands would necessarily be increased with a focus on design, management and maintenance that reduces mosquito populations to below threshold treatment levels.

2.6 Alternatives Summary

Table 3 below summarizes the activities that would be permitted on the Great Meadows Unit at the Refuge under each of the mosquito management alternatives.

Table 3. Summary of mosquito management activities permitted under each alternative.

2 Alternative Monitoring and Pesticides Permitted for Use Restore Access Surveillance of Tidal 1 Mosquito Populations Circulation 3 A Yes L, P, A Yes Limited in sensitive species habitat and along tidal channels and sloughs. B Yes L (Bti and Bs only) Yes Same as A C Yes Pesticides not permitted Yes Same as A 1 See Appendix I for methods 2L = larvicides (Bti and Bs), P = pupacides (monomolecular film), A = adulticides (pyrethrin or pyrethroids) 3 Phase 5 = High risk of mosquito-borne disease

Chapter 3 AFFECTED ENVIRONMENT

3.1 Physical Environment

3.1.1 Climate The Connecticut Coast lies in the humid zone of the temperate climate range and experiences warm summers and cold winters. The climate is influenced year-round by the moderating effects of the Atlantic Ocean and Long Island Sound. According to NOAA’s 100 year averages (1901-2001), the average daily summer temperature is 68.5 degrees Fahrenheit and the average daily winter temperature is 27.5 degrees Fahrenheit (NOAA State Annual and Seasonal Time Series website). Annual precipitation averages 47 inches, with approximately 39 inches of snowfall each year. Thunderstorms occur on an average of 22 days each year, primarily during the summer months (USFWS 1989a).

3.1.1a Climate Change in Connecticut.From 2011 through 2016, The Nature Conservancy (TNC) worked with Connecticut’s coastal communities to assess local vulnerability to sea level rise and storm surge impacts, as well as identify unprotected parcels of land that would accommodate the predicted salt marsh advancement, using the Coastal Resilience Tool (http://coastalresilience.org/project-areas/connecticut- solutions/, accessed September 2016). The Coastal Resilience Tool allows users to visualize various seal level rise and/or storm surge scenarios, analyze the potential impacts on communities, natural resources, and critical infrastructure such as roads and schools at the parcel scale (i.e., a 0.61-meter raster cell size in a GIS). Connecticut is now the first state in the nation to complete such a detailed assessment down to the parcel scale. As a result of this work, coastal communities in Connecticut can more effectively work together to promote the ecological and socioeconomic health of their communities. Furthermore, TNC’s Coastal Resilience Tool is a user-friendly, internet-based tool that allows its users to visualize various predicted climate change scenarios that have been adopted by Connecticut’s local and State government planning, including the 2015 WAP (CT DEEP 2015a).

3.1.1b Climate Change and Mosquito Management. “Unfortunately, rising temperatures and other changes in our weather we will see as a result of climate change increases the likelihood of tropical mosquito-borne diseases gaining a foothold in Connecticut, so it is critical for us to continue building on effective strategies we have put in place to control these insects and to increase public awareness of this challenge.” - DPH Acting Commissioner Dr. Raul Pino (regarding the development of a plan for preparedness for the Zika virus) 27

As described in Dr. Raul Pino’s quote above, rising temperatures associated with climate change may alter the types of mosquitoes and mosquito borne diseases that occur in Connecticut. Some diseases that we may have to look out for in the future because of rising temperatures are Zika, dengue, chikungunya, and malaria. Another aspect of climate change is sea-level rise. The changes in the rise of the water may alter the hydrology of our coastal marshes and lands adjacent to the Refuge’s salt marshes. The sites that are now mosquito breeding areas could change due to inundation of salt water or other factors associated with sea- level rise. The Refuge bases its predictions for sea-level rise using TNC’s Coastal Resilience Tool as addressed above.

3.1.2 Topography, Geology, and Soils The Connecticut Coast was shaped by gradual submergence which produced a series of flooded valleys and a fringe of islands, peninsulas, channels and bays. Except for the lowlands of the Connecticut River Valley, the coast is made up of gently rolling hills and rock outcrops, known as the Southern New England Uplands. Upland soils are composed chiefly of sands, gravels, and clays with fertile loam depressions.

Glacial deposits comprise the islands and barrier beaches currently found along the Connecticut coast. Glacial sediment carried down the Housatonic River and deposited at its mouth created sand and mud flats along the coast of Stratford.

Coastal sand and mud flats provide ideal sites for salt marsh formation. Marsh substrate consists of gravel and sandy soils overlain with organic peat soils grading to silt in some areas. Tidal marsh soil is classified as Westbrook Mucky Peat (USFWS 1989a).

Nearly level to moderately steep loamy soils characterize the coastlines of Stratford and Bridgeport. In 1955, the Army Corp of Engineers used material contaminated with heavy metals and dredged from Bridgeport Harbor to construct dikes within the Great Meadows Unit (GMU). The Service studied organochlorine and heavy metal levels during a Level II Pre-Acquisition Survey, and further analyzed sediment for chromium, pesticides, chlorinated herbicides, semivolatile organics, ETPH, PCBs, TOC, and grain size. Both studies concluded levels to be far below threshold concentrations, and therefore do not pose a threat to the public or to wildlife.

3.1.3 Air Quality The State of Connecticut Department of Energy and Environmental Protection (CT DEEP) and its predecessor departments have monitored some aspects of air quality since the 1950s. The Federal Clean Air Act of 1970 (CAA), as amended, established the USEPA, which implements National Ambient Air Quality Standards (NAAQS) for six criteria pollutants: carbon monoxide, lead, nitrogen dioxide, ozone, sulfur dioxide, and particulate matter. These pollutants are considered harmful public health and welfare, and detrimental to sustaining healthy ecosystems. The CT DEEP has also monitored for toxic pollutants and metals including volatile organic compounds, aldehydes, polycyclic aromatic hydrocarbons, mercury, and dioxin. The CT DEEP ambient air monitoring network consists of 20 monitoring stations. Although no air quality monitoring stations are located within the Refuge boundaries, 8 stations are situated within 20 miles of Refuge management units.

The NAAQS establish air quality attainment goals for each of the six criteria pollutants, expressed as maximum concentrations measured in parts per million or parts per billion (ppm or ppb). Concentrations of all six criteria pollutants have been generally decreasing throughout Connecticut (CT DEP 2010): • from the mid-1970s to the present for ozone, carbon monoxide, nitrogen dioxide, and sulfur dioxide; • from the mid-1970s until monitoring in Connecticut was discontinued 2002 for lead; and • from 2001 to present for particulates.

Sources of air pollution in Connecticut include both mobile sources (primarily motor vehicles) and stationary sources (such as power plants and industrial facilities). The greatest concentration of air pollution sources in the 28

Refuge vicinity is New York City, located approximately 25 miles southwest from the Refuge’s Calf Island Unit in Greenwich.

3.1.4 Water and Water Quality Great Meadows Marsh and Lewis Gut are located within the Great Meadows Marsh drainage sub-basin, within the Southwest Coast Basin, and west of the Housatonic River Basin. Erosion of the main headland to the east of Great Meadows formed the Long Beach peninsula. The protection of Long Beach by the back-barrier lagoon, Lewis Gut, encouraged the formation of the Great Meadows salt marsh complex.

The Lewis Gut system once supported 1,450 acres of tidal wetlands, of which only about a third remains today. The integrity of the hydrological regime and water quality of the Great Meadows system have been impacted throughout the years by land use changes and urban development, such as the construction of the Bridgeport- Sikorsky Airport, Lordship Boulevard, landfills, and the disposal of dredged material for industrial, commercial, and residential areas (USFWS 2001).

Sources of freshwater to the Lewis Gut embayment include a ditch extending from Frash Pond to the GMU and storm water runoff from adjacent areas. Prior to significant urban development of the past century, Neck Creek connected Lewis Gut to the Housatonic River. Due to extensive tidal flushing, Lewis Gut is strongly influenced by the water quality of Bridgeport Harbor and Johnsons Creek. Both areas experience periodic high counts of coliform bacteria (King’s Mark 1987). Heavy industrial and residential development contributed to the poor sediment quality, the potential for contamination from heavy metals, and low dissolved oxygen levels of Bridgeport Harbor (CT DEEP 1998). Salinity measures taken from Lewis Gut range from about 18 ppm to 26 ppm (Integrated Waterbird Management and Monitoring survey; IWMM).

Historical aerial photographs reveal three ponds on the Great Meadows Unit. Two are currently dominated by open water and the third is a depression dominated by common reed (Phragmites australis). In 2001, Service staff found the salinity of the largest pond at GMU (2.72 acres) to be 2-3 parts per thousand (ppt). Water temperatures of this pond during winter vary from of -0.1C at the surface to 3.0C near the bottom. The low salinity may be a result of saltwater intrusion through storms and dike breeches or salt leaching from the surrounding dredge spoil (USFWS 2001).

The smallest pond (0.70 acres) had a salinity of 11-12 ppt at the surface and 20 ppt in February and 10 ppt on the surface in July of 2001. Although trivalent chromium occurs in sediment of the ponds below threshold concentrations, precautions should be taken to avoid direct dermal contact by humans to prevent skin irritation (www.osha- slc.gov/SLTC/healthguidelines/ chromium3/recognition.html). The ponds are located in areas closed to the public.

3.2 Biological Considerations The Refuge provides a variety of environments, each with its own characteristic set of flora and fauna. Environments throughout Stewart B. McKinney NWR have been altered by past and current human actions including human development, agriculture, and other land uses. Today, land managers of remaining open spaces and interested partners are working towards enhancement and restoration of the terrestrial and wetland environments of the Refuge. These efforts provide opportunities to enhance or expand existing habitats for the benefit of wildlife, plants, and people. An important consideration as we move forward is to ensure that our actions do not enhance or create conditions in which mosquito populations increase above levels that are naturally found in these wetland environments.

3.2.1 Vegetation and Habitat Types The Great Meadows Marsh system, which includes properties owned the Service, the City of Bridgeport, and the Town of Stratford, is comprised of tidal salt marsh, filled wetlands and upland, barrier beach, and the Lewis Gut embayment. This 600 acre complex is a remnant of what was once an extensive tidal-marsh system covering approximately five square miles extending from Johnsons Creek in the west to the Housatonic River in the east 29

(King’s Mark ERT 1987). The Service’s Northeast Coastal Areas Study (1991) identifies this area as the Lower- Housatonic River-Great Meadows Marsh Complex, an important coastal habitat site containing the largest block of unditched high salt marsh (225 acres/91 ha) in Connecticut.

Salt marsh vegetation is organized along a gradient depending on species tolerance for saline and anoxic conditions (Mitsch and Gosselink, 2000). The low marsh, which receives tidal flow twice daily, is generally dominated by salt marsh cord grass (Spartina alterniflora). The high marsh, which receives less regular tidal influence, is comprised of a variety of species including salt marsh hay (Spartina patens), black grass (Juncus gerardii), and spike grass (Distichlis spicata) (Nixon, 1982; Bertness, 1991). Salt pannes, pools and tidal creeks may also characterize salt marsh habitats. Salt pannes are bare, exposed, or water-filled depressions in a salt marsh (Mitsch and Gosselink, 2000). Most of the Great Meadows marsh is low marsh.

According to the State Plant Ecologist, there are three state listed species that occur on the Great Meadows Unit. They are beach needle grass (Aristida tuberculosa – state endangered), bayonet grass (Bolboschoenus maritimus ssp. Paludosus – state species of concern), and salt marsh pink (Sabatia stellaris – state endangered). In fact, the population of salt marsh pink at Great Meadows is the last natural population of this plant in the state.

3.2.1a Salt Marsh Pink. Salt marsh pink, Sabatia stellaris, is a state endangered annual plant that grows on open, sandy soils at the upper edges of salt and brackish marshes. Although salt marsh pink is abundant along much of the Atlantic and Gulf coast, it is rare in New England and only exists at one site in Connecticut, which is on the Refuge. In partnership with the State Plant Ecologist and the New England Wildflower Society, the population on the Refuge is regularly mapped and monitored. Seed has also been collected from the site.

One of the biggest threats to this species in New England and on the Refuge is the spread of the invasive plant species, Phragmites australis. Salt marsh pink requires open space and cannot compete with this tall invasive grass. The population of salt marsh pink at the Refuge site has declined since 1992 when over 10,000 individuals were found in several large areas. Management for this species would require opening up sandy space adjacent to present populations at the site by controlling Phragmites using mechanical and chemical methods, as well as minimizing human disturbance and the development of wrack in these areas.

3.2.2 Special Biological Communities/Critical Wildlife Habitat The Refuge was established to conserve habitats of migratory birds such as wading birds and shorebirds, as well as federally endangered and threatened species such as the roseate tern (Sterna dougallii) and the piping plover (Charadrius melodus). The Refuge also provides breeding or wintering habitat for many state-listed species. Table 4 provides a list of federal or state listed species that occur or have the potential to occur on the Refuge.

3.2.3 Noxious Weeds/Exotic Plants Non-native and exotic plants exist in virtually every habitat type on the Refuge. Special attention must be paid to any mechanical manipulation of wetlands for mosquito control to prevent the development of conditions that encourage colonization and spread of invasive plants. In some instances, the use of mechanical means to control mosquito populations may help reduce and/or eliminate invasive weeds by improving tidal inundation cycles.

Dominant invasive plants of the salt marsh habitats on the Refuge include common reed (Phragmites australis), princess tree (Paulownia tomentosa), perennial pepperweed (Lepidium latifolium), and spotted knapweed (Centaurea maculosa). Common reed occupies areas around the ponds, as well as areas that have altered hydrology and are upland. Much of the upland consists of common reed. Princess tree and spotted knapweed are found on the dikes and higher upland areas such as the railroad tracks. Although the Great Meadows Unit does not have perennial pepperweed as of yet, this invasive plant is in the state so the potential for invasion is there.

Table 4: Federal and State listed wildlife and plant species that occur or have the potential to occur on the Stewart B. McKinney NWR

3 Occurrence

Common Name Refuge T&E Federal T&E State LANDBIRDS American Kestrel X SC

Bald Eagle X T

Barn Owl E

Broad-winged Hawk X SC

Brown Thrasher X SC

Cerulean Warbler X SC

Common Nighthawk X E

Horned Lark X E

Ipswich Savannah Sparrow X SC

Long-eared Owl X SC

Northern Harrier X E

Northern Parula X SC

Northern Saw-whet Owl X SC

Peregrine Falcon X T

Purple Martin X SC

Red-headed Woodpecker X E

Saltmarsh Sparrow X SC

Seaside Sparrow X T

Sedge Wren X E

Sharp-shinned Hawk X E

Short-eared Owl X T

Whip-poor-will SC

WATERBIRDS American Bittern X E

Common Loon X SC

Common Tern X SC

Glossy Ibis X SC

Great Egret X T

Least Bittern X T

Least Tern X T

3 Occurrence

Common Name Refuge T&E Federal T&E State Little Blue Heron X SC

Pied-billed Grebe X E

Snowy Egret X T

Yellow-crowned Night Heron X SC

SHORE AND SEA BIRDS American Oystercatcher X T

Piping Plover X T T Red Knot X T

Roseate Tern X E E Upland Sandpiper E

MAMMALS Eastern Red Bat X SC

Eastern Small-footed Bat X E Harbor Porpoise SC

Harbor Seal X

Hoary Bat SC

Least Shrew E

Little Brown Bat X E

Northern Long-eared Bat T E

Silver-haired Bat SC

Southern Bog Lemming SC

Tri-colored Bat E

REPTILES AND AMPHIBIANS Atlantic Hawksbill E

Eastern Box Turtle X SC

Eastern Ribbonsnake SC

Green Sea Turtle T T

Kemp's Ridley Turtle E E

Leatherback E E

Loggerhead Sea Turtle E T

Northern Diamond-backed Terrapin X SC

Smooth Green Snake X SC

Wood Turtle SC

FISH Atlantic Sturgeon E E

3 Occurrence

Common Name Refuge T&E Federal T&E State Blueback Herring X SC

Sea Lamprey X

Shortnose Sturgeon E E

Spiny Dogfish X

Striped Bass X

Summer Flounder X

INVERTEBRATES American Burying Beetle E SC EX

Atlantis Fritillary X E

Northeastern Beach Tiger Beetle T SC EX

Puritan Tiger Beetle T E

Saltmarsh Tiger Beetle SC PLANTS Bayonet Grass X SC

Beach Needle Grass X E

Blazing-star SC

Dillenius' Tick-trefoil SC

Dioecious Sedge SC

Eastern Prickly-pear X SC

Featherfoil SC

Fragrant Sumac SC

Golden Alexanders E

Hairy Forked Chickweed SC EX

Lilaeopsis SC

Marsh Pink X E

Mudwort SC

Panic Grass X T

Parker's Pipewort E

Red Goosefoot SC EX

Seabeach Sandwort X SC

Sickle-leaf Golden-aster X E

Small Skullcap E

Smooth Black-haw X T

Starry Campion T

Stiff Goldenrod E

3 Occurrence

Common Name Refuge T&E Federal T&E State Swamp Cottonwood T

Tall Cinquefoil SC

Yellow Pimpernel E

Yellow Thistle E

1 Refuge Occurrence X=Species is known to occur on the Refuge, as provided by several physical surveys, observations, and inventories. 2 Federal T&E Federal Endangered Species List. E-Endangered; T-Threatened. (https://www.fws.gov/endangered) 3 State T&E Connecticut’s Endangered, Threatened, and Special Concern Species-2015. E-Endangered; T-Threatened; SC-Special Concern; EX- Believed Extirpated. (CT DEEP 2015b)

3.2.4 Wildlife Located along the Atlantic Flyway, the Great Meadows Unit is an important site for migratory birds including waterfowl, shorebirds, wading birds, raptors, and passerines. The GMU was recognized in the Atlantic Coast Joint Venture Plan of the North American Waterfowl Management Plan (USFWS 1989b). Over 270 species of birds have been observed on or near the area since 1900 (King’s Mark 1987). In the 1920s, Roger Tory Peterson, Allan Cruickshank and other prominent ornithologists regularly traveled by train from New York City to Stratford Great Meadows to birdwatch, then the premier habitat in the entire greater metropolitan New York region (TNC 1993). It has also been classified as an Important Bird Area by Audubon Connecticut. See http://ct.audubon.org/conservation/stratford-great-meadows-area for more information.

A National Diversity Database request of state-listed species that may be found at the Great Meadows Unit gave the following information:

Scientific Name Common Name State Status Vertebrate Animal Ammodramus caudacutus Saltmarsh sharp-tailed sparrow SC Ammodramus maritimus Seaside sparrow T Asio flammeus Short-eared owl T Botaurus lentiginosus American bittern E Circus cyaneus Northern harrier SC Ixobrychus exilis Least bittern T Gallinula chloropus Common moorhen E Malaclemys terrapin terrapin Northern diamondback terrapin SC Podilymbus podiceps Pied-billed grebe E Rallus elegans King rail E Invertebrate Animal Cicindela marginata Saltmarsh tiger beetle SC Vascular Plants Aristida tuberculosa Beach needle grass E

Bolboschoenus maritimus ssp. paludosus Bayonet grass SC

Sabatia stellaris Marsh pink E

3.2.4a Waterfowl. The Great Meadows marsh and Lewis Gut are important feeding and staging areas during winter storms for Canada Geese (Branta canadensis), American black ducks (Anas rubripes), mallards (Anas platyrhynchos), and diving ducks such as scaup (Aythya spp.), common goldeneye (Bucephala clangula), bufflehead (Bucephala albeola), and old squaw (Clangula hyemalis) (USFWS 1989a). Species which have historically nested at Great Meadows include pied- billed grebe (Podilymbus podiceps), gadwall (Anas strepera), black duck, mallard, green-winged teal (Anas crecca), blue-winged teal (Anas discors), northern shoveler (Anas clypeata), mergansers (Mergus merganaser), and bufflehead (King’s Mark 1987 and USFWS 1989a). Waterfowl and shorebirds heavily utilize the tidal mud flats and the high marsh when feeding.

The Connecticut DEEP has conducted statewide midwinter waterfowl (aerial) surveys in coordination with the annual Service national census. Areas surveyed include the coastal region of Connecticut and major rivers and selected lakes within ten miles of the Long Island Sound. Dabbling ducks include mallard, American black duck, gadwall, American wigeon, green-winged teal, northern shoveler, and northern pintail. Diving ducks include redhead, canvasback, scaup, ringneck duck, common goldeneye, bufflehead, ruddy duck, eider, scoter, long-tailed duck, and merganser (USFWS 2007 and CT DEP 2007).

The Integrated Waterbird Management and Monitoring surveys were conducted at Great Meadows from 2011 to the present. Some of the most common species >100 (count for high year) are American black duck (323 – 2013), brant (522 – 2012), and Canada goose (229 - 2014). Other waterfowl species seen include 35

American widgeon, northern pintail, green-winged teal, blue-winged teal, bufflehead, common merganser, Eurasian widgeon, gadwall, hooded merganser, mallard, northern shoveler, red-breasted merganser, and ring-necked duck.

3.2.4b Shorebirds/Seabirds. Protected or undisturbed portions of Long Beach support nesting populations of piping plovers (Charadrius hiaticula), American oystercatchers (Haemotopus palliatus), common terns (Sterna hirundo), least terns (S. antillarum), killdeer (Charadrius vociferus), and spotted sandpipers (Actitis macularia).

More than 5000 individual shorebirds roost on backdune sandflat communities of Pleasure and Long beaches during migration (USFWS 1991). Nesting and migrating shorebirds, gulls, and terns utilize the Great Meadows marsh as a feeding and loafing area. Wintering residents include herring gulls, great black backed gulls, and double crested cormorants.

The following species have been identified during the IWMM surveys – sanderling, semipalmated plover, black-bellied plover, dunlin, least tern, spotted sandpiper, semipalmated sandpiper, short-billed dowitcher, lesser and greater yellowlegs, killdeer, least sandpiper, and common tern.

3.2.4c Marsh Birds. There are records of both seaside (state threatened) and saltmarsh sparrow (state species of concern) nesting at Great Meadows (Kruitbosch and Leenders, 2009). They were predominately found near Area D – area of public health concern identified by the CT MMP in 2014 and in the marsh just east of the FedEx buildings. Additionally, the annual rail surveys and the bird surveys associated with the Salt Marsh Integrity Index have shown clapper rail and willet using the marsh for breeding throughout the site. These species nest on the ground or close to the ground, which makes their nests susceptible to trampling or spring flooding tides.

Clapper Rail - According to the Saltmarsh Habitat & Avian Research Program (SHARP) study results, the clapper rail has seen significant declines at about 12.9% annually since 1998 in Connecticut and about a 4.2% decline annually in the region (USFWS Region 5). This may be due to habitat loss from changes in marsh hydrology, limits on sedimentation deposits, and rises in sea- level. The clapper rail is listed as a “high priority” species in BCR 30 and is in the State’s Wildlife Action Plan, as a “very important” species of greatest conservation need.

Salt marsh Sparrow - This saltmarsh specialist is one of 19 species on the Partners in Flight Landbird Conservation Plan’s Red Watch List. The species has lost 94% of its global population over the last 44 years (1970-2014) due to climate change (sea-level rise) and urbanization (development) and continues to decline 9.0% annually (Correll et al. 2016). In fact, Correll et al. predicted that within the next 50 years if nothing changes, the species population will likely collapse and extinction will be inevitable. The authors urge “immediate conservation actions” and recommend restoring sediment to salt marshes, which would increase the amount of high marsh for breeding habitat for species like the saltmarsh sparrow.

Seaside Sparrow - The studies conducted by SHARP detected no changes in the breeding population of this species in Connecticut. It is, however, a State threatened species and on the Partners in Flight “yellow watch list”, which means that “species not declining but vulnerable due to small range or population and moderate threats”.

3.2.4d Mammals.At least 24 species of mammals have been observed on the Refuge, including six species of bat, five of which are state listed species (2 endangered and 3 species of concern). Most of these observations were made with trail cameras or by tracking species. All bat “observations” were acquired using a bat monitor that records echolocation sounds. No bat data has been obtained from the Great Meadows Unit. Additionally, an intensive mammal survey that includes small mammals has not been 36

undertaken as of this date. Therefore, there are likely more species of mammal, especially small mammal, that occupy habitat on the Refuge. A CT National Diversity Database request (March 2017; see list above) did not reveal any mammal species of concern with the potential to occur at this site.

3.2.4e Reptiles and Amphibians.Although extensive reptile and amphibian surveys have not been conducted on the Refuge, the Refuge staff knows of at least ten species that utilize the habitat at Stewart B. McKinney. These species include black rat and garter snakes, red-backed salamanders, spring peepers and wood frogs. Two of the turtles that are known to occupy Refuge property are species of concern in the state – the northern diamondback terrapin and the eastern box turtle. The National Diversity database request identified the northern diamondback terrapin as the only reptile of concern that may occur at the Great Meadows site.

Northern Diamondback Terrapin – Northern diamondback terrapins (Malaclemys t. terrapin), nest at Great Meadows and are found in large numbers in the tidal creeks of the unit. Diamondback terrapins have a gray, light brown or black carapace that is broad, wedge-shaped, and patterned with concentric rings or ridges. The plastron can range from yellowish to greenish gray and their head, tail, and legs are usually light gray and spotted black. This is the only species of turtle in North America that spends its life in brackish water. There are sandy areas adjacent to the marsh where female turtles nest.

3.2.4f Fisheries.The Housatonic River supports important anadramous fish runs for American shad (Alosa sapidissima), sea-run brown trout (Salmo trutta), alewife (Alosa pseudoharengus), blueback herring (Alosa aestivalis), striped bass (Morone saxatilis), white perch (Morone americana) and Atlantic sturgeon (Acipenser oxyrhynchus) (USFWS 1991). Topminnows and killifish (Fundulus spp.) were found in the Great Meadow ponds during site visits, with possible sticklebacks (Family Gasterosteidae), sheepshead minnows (Cyprinodon variegatus) and eels in the small pond (USFWS 2001). For a complete list of finfish species found in the waters near the Great Meadows Unit of SBMNWR see Table 5 below.

3.2.4g Invertebrates.Invertebrates are considered an important component of any habitat, including tidal ecosystems. Despite their importance to ecosystems as a whole, little is known about the ecology and biology of invertebrates (excepting mosquitoes) within the Great Meadows Unit of the Refuge.

The mouth of the Housatonic River contains important natural shellfish beds, particularly for American oysters (Crassostrea virginica) and hard-shelled clams (Mercenaria mercenaria). Although invertebrate surveys of the Great Meadows Unit have not been conducted, typical salt marshes most likely characterize the Great Meadows. According to the Tidal Wetland Ecology of Long Island Sound (Parts 2 and 3), the ribbed mussel (Geukensia demissa), marsh fiddler or black fiddler (Uca pugnax), red-jointed fiddler (Uca minax), marsh crab (Sesarma reticulatum), striped sea anenome (Haliplanella luciae), common clamworm (Nereis succinea), rough periwinkle (Littoria saxatilis), and mud snail (Ilyanassa obsoleta) can be found in the low marsh, while the saltmarsh snail, saltmarsh isopod (Philosocia vittata), and saltmarsh amphipod (Orchestia grillus and O. uhler) are found in typical high salt marsh ecosystems.

Saltmarsh Tiger Beetle – The 2017 National Diversity Database request identified the saltmarsh tiger beetle as an invertebrate species of concern that may occur at the Great Meadows Unit. This species is a brown-bronzed to green-bronzed beetle with metallic green below. They are found most often on mudflats in salt marshes on the bay side of barrier beaches in all New England states except for Vermont. The saltmarsh tiger beetle can be seen in the northern regions form July to mid- September. The burrows for this species are in sandy vegetated soil and are approximately 25 centimeters (cm) deep. Not much is known about their life cycle (Leonard and Bell 1999).

Table 5. Finfish species found in the waters near the Great Meadows Unit of SBMNWR

Long Beach/Lewis Gut Seasonal Finfish Occurrence* Common Name Scientific Name Area Jan-Mar Apr-Jun Jul-Sep Oct-Dec Alewife (Alosa pseudoharengus) G A S A,J A,J Blackfish (Tautoga onitis) B,G - S S,J A,J Bluefish (Pomatomus saltatrix) 0 - A A,J A,J Butterfish (Peprilus triacanthus) B - A A - Cunner American (Tautogolabrus adspersus) B,G A S S,J A,J Eel (Anguilla rostrata) G - A A A Summer Flounder (Paralichthys dentatus) B - A A - Winter Flounder (Pseudopleuronectes americanus) B,G S S A,J A,J Windopane Founder (Scophthalmus aquosus) B,G A S S,J A,J Killifish (Fundulus spp.) G A S S,J A,J Mackerel (Scomber scombrus) B,G - A A - Menhaden (Brevoortia tyrannus) B,G - A A - Pipefish Rainbow (Syngnathus fuscus) B,G S S,J A,J A,J Smelt Sand Lace (Osmerus mordax) B,G S S,J J J Scup (Ammodytes americanus) B,G - A A S Sheepshead Minnow (Stenotomus chrysops) B,G - S S - Silversides (Cyprinodon variegatus) G A S S,J A,J Sticklebacks (Menidia menidia) G A S S,J A,J Striped Bass (Apeltes spp., Gasterosteus spp.) G A S S,J A,J Tomcod (Morone saxatilis) B,G - A A A Weakfish (Microgadus tomcod) G S, J A A,J S,J White Perch (Cynoscion regalis) (Moron B - S S,J A,J americanus) G A S A,J A,J

Areas: B - Long Beach Area Occurrence: A - Adults (>=1 year) G - Lewis Gut / Great Meadows S - Spawning adults J - Young of the year juveniles - - Not present

*Table 3, Page 61. King's Mark Environmental Review Team. 1987. Environmental Review Team Report: Long Beach, Stratford, CT. Wallingford (CT): King's Mark Resource Conservation and Development Area, Inc. 83 pp.

3.2.4h Mosquitoes.Mosquitoes are typical nematoceran dipterans with aquatic immature stages and aerial adult stages. Eggs must come in contact with water in order to survive. Mosquitoes have four larval stages (instars) and one aquatic pupal stage. The aerial adult emerges from the pupal stage onto the surface of the water, expands its wings, hardens its exoskeleton, and flies off. The biology, diseases and pest ability of each mosquito species is different and influences decisions concerning control strategies.

A brief summary of the most common mosquito species detected on the Refuge are presented below. A more detailed account of mosquito biology and vector capabilities within the region is presented in Appendix J.

Aedes cantator – otherwise known as Ochlerotatus cantator, has a distribution that extends from Labrador, Newfoundland and Nova Scotia south along the east coast of the United States to Virginia. Isolated populations have been reported from some inland areas east of the Mississippi River. The mosquito reaches greatest abundance along the Atlantic coast of New Jersey and areas that border Delaware Bay. Aedes cantator is a multivoltine species that occurs in greatest numbers during early spring. The initial generation of larvae can usually be found along the upland edge of salt marsh habitats during April. Larval populations from the spring brood generally peak by mid- May and become mixed with those of Aedes sollicitans. As the season advances, larvae appear in lesser numbers but become distributed over a wider range of salt marsh habitat. Larvae can generally be collected from both salt and brackish habitats well into the fall. Aedes cantator is recognized as a salt marsh species, but the larvae can be found in a wide range of floodwater habitats. The species is most common in mixed stands of Spartina patens, Spartina alterniflora and Distichlis spicata along the upland edge of salt marsh habitats. Aedes sollicitans, Culex salinarius and Anopheles bradleyi are the most common species associated with salt and brackish water habitats.

Aedes sollicitans – The eastern saltmarsh mosquito, otherwise known as Ochlerotatus sollicitans, is a species of mosquito native to the eastern seaboard of the United States and Canada as well as the entire Gulf coast and is also present in the Bahamas and Greater Antilles. While primarily found in coastal areas within a few miles of the coast, it is occasionally found inland in areas with saline pools; the species was reported as far west as Arizona. The species is a prime vector for Eastern equine encephalitis (EEE), Venezuelan equine encephalitis (VEE), and dog heartworm. Aedes sollicitans has a conspicuous band of white scales around the central area of the proboscis and the anterior portion of the hind tarsomeres upon which there is also band a band of yellow scales in the middle. The abdomen has white basal bands and is divided by a medial longitudinal stripe. The thorax is white on the sides and the top is brown, yellow, golden, and white (iNaturalist Eastern Saltmarsh Mosquito description). The female Aedes sollicitans lays her eggs on the dried out substrate of salt pannes, depressions within salt marshes that dry out between periods of very high tides (spring tide). Spring high tides fill the panne to create optimal conditions for the eggs to hatch. The eggs hatch in 4-5 days with optimal conditions upon the panne filling at the next spring tide. Aedes sollicitans tends to stay within 5 miles of the coast on average all the range can be greater dependent upon a number of factors such as wind speed and duration. It tends to feed most actively at twilight but is an opportunistic feeder and will feed on a host species that enters its area in daytime. The female requires one blood meal for each egg batch with the primary host species being mammals, and birds as a secondary host. In the south the peak number of adults occurs in the spring and fall, and in the northern portion of its range peak adult population occurs in the summer. The last batches of eggs laid in the fall remain in diapause until the spring.

Aedes taeniorhynchus – The black salt marsh mosquito, otherwise known as Ochlerotatus taeniorhynchus, is very common in the eastern coastal areas of the Americas. Although it is not a primary vector of major concern, it can transmit pathogens to humans and other animals. Its characteristic emergence in large numbers after rains and flooding events as well as its aggressive biting contribute to its notoriety as a pest insect (CAES 2005). This species is present in all of the coastal counties in Connecticut and inhabits high-tide salt marsh pools in salt marsh habitat. Its host preference is mammals and can be found in mostly shady locations or at night. This species is medically relevant, primarily as a vector of two alphaviruses from the family Togaviridae, EEE and (VEE (Drew 2001). Additionally, black salt marsh mosquitoes have also been known to transmit the filarial worm Dirofilaria immitis, commonly known as the dog heartworm (University of Florida).

Other species of mosquitoes – Additional species that may be found at the Great Meadows Unit of the Refuge are Culex pipiens, Cu. restuans, Cu. salinarius (prefers brackish water), Anopheles punctipennis, An. Quadrimaculatus, Aedes vexans, possibly Ae. albopictus (Asian tiger mosquito - the Ag Exp Station has found it in other parts of Stratford/Bridgeport).

3.3 Socioeconomic Considerations

3.3.1 Cultural Resources The Great Meadows Unit was used by both Native Americans and early European settlers. Decades before the arrival of the first settlers in 1639, Native Americans inhabited the Johnsons Creek area each summer and actively used the marsh for fishing, oystering, clamming, and hunting game birds. There was also a large Paugussett village near Frash Pond, just north of Great Meadows (Doucette and Elam 2011).

Rich game, fish, shellfish and other natural resources in and around the Great Meadows Unit supported seasonal and permanent settlements. Productive oyster beds supported a historically important industry and abundant salt marsh hay offered open grazing pasture for cattle and material for roof thatching. Other practical uses for marsh and beach plants included candles, jelly, tea, and seasonings for food and salad. The 20th century saw the marsh used for waterfowl hunting and recreation.

Although the Great Meadows marsh has a significant human history, the 2011 Archaeological Overview Assessment for the Refuge makes it clear that the “low-lying, waterlogged terrain and the poorly drained soils do not lend themselves to human habitation.” Therefore the prospect of archaeological or post-contact cultural resources being found in the wetland areas, which were used by humans for hunting, fishing and recreation - but not for settlement, has a low to moderate probability (Doucette and Elam 2011).

3.3.2 Socioeconomics and Environmental Justice Although Connecticut is the third smallest state in the nation, it has the fourth greatest population density at 738 persons per square mile (USCB 2010). Within the state, population density is highest in Fairfield County, which is where the Great Meadows Unit is located. The Connecticut Department of Energy and Environmental Protection classify Stratford and Bridgeport, the two towns where the Great Meadows Unit is found, as “Urban Core” communities. The combined population of the two towns is approximately 200,000.

The Great Meadows Unit consists largely of marsh with a small amount of upland. The land did not historically and does not now serve as housing or locations of business for the local population. However, there are large industrial and warehouse buildings employing many workers located directly adjacent to the Refuge boundary. Minority or low income populations inhabit housing in areas within 1-2 miles of the Refuge boundary. The Great Meadows Unit of the Refuge provides the local populace and local workers with many benefits, including an accessible green space, walking trails for exercise, wildlife viewing areas, and a place to hunt waterfowl. These are some of the activities identified in the Connecticut Statewide Comprehensive Outdoor Recreation Plan as in high demand and likely to increase. Therefore, the Refuge is a part of a larger patchwork of publicly-owned lands that can help to achieve equal access to natural areas across socioeconomic groups.

Mosquito management and control would be especially beneficial to the local populace and local workers surrounding Great Meadows, as they are the most likely to be negatively affected by a large mosquito population and potential disease near their homes and places of employment. The anticipated effects of each of the alternatives of this plan would occur only within the boundaries of the Refuge and do not involve loss or acquisition of businesses, residential homes or community facilities.

3.3.3 Land Use The Refuge was established to for the following purposes: • To enhance the populations of herons, egrets, terns, and other shore and wading birds within the Refuge; • To encourage natural diversity of fish and wildlife species within the Refuge; • To provide for the conservation of all fish and wildlife within the Refuge; • To fulfill the international treaty obligations of the United States respecting fish and wildlife; and • To provide opportunities for scientific research, environmental education, and fish and wildlife- orientated recreation.

The Refuge consists of about 1,000 acres of island and mainland habitat. The Service manages the Refuge consistent with the purposes for which the Refuge was established and the mission of the National Wildlife Refuge System. Significant public laws, regulations and policies that assist in Refuge management are described in section 1.5.3. Other than Refuge approved recreational activities and operation and maintenance activities, no other land use exists on the Refuge. Management of the Great Meadows Unit has centered on protecting and enhancing salt marsh habitat for resources of concern such as saltmarsh sparrows and salt marsh pink.

3.3.3a Public Use of the Refuge. Public use of the Refuge is limited to wildlife-dependent recreational activities, or the Big 6, which includes hunting, fishing, wildlife photography, wildlife observation, interpretation, and environmental education. Approximately 27,000 people visit the Refuge annually for these wildlife dependent activities. The Great Meadows Unit has a small trail system, as well as a designated waterfowl hunting zone. There is no fishing on the Refuge as of this date. Access is provided through SUPs and partnerships for certain types of research which is conducted throughout most of the year. Mosquito control activities must be adequately coordinated with the Refuge Manager to avoid applying mosquito control products, such as pesticides, when visitors, partners, and staff are present.

3.3.3b Surrounding Land Uses.The Refuge spans 70 miles of the Connecticut coast, which is the most densely populated area in the state. In fact, Connecticut is the fourth most densely populated in the United States predominantly due to the coastal population. About 200,000 people live in the towns and cities surrounding the Great Meadow Unit (Town of Stratford and the City of Branford). Most of the land use around the unit is industrial and commercial with some residential zones bordering the eastern side of the marsh.

3.3.3c Human Health and Safety Concerns. The management of mosquitoes in Connecticut is a collaborative effort involving the DEEP, CAES and the DPH, together with the Department of Agriculture and the Department of Pathobiology at the UCONN. These agencies are responsible for monitoring and managing the state’s mosquito population levels to reduce the potential public health threat of mosquito-borne diseases.

The mosquito control program was created by a legislative act (PA 97-289) in 1997 to monitor and control the spread of EEE, a potentially deadly disease. The EEE is a virus that is present in nature and is cycled in the wild bird population by certain species of bird-feeding mosquitoes. The virus has no effect on wild birds; however, it can be fatal to humans, horses and commercial exotic fowl (e.g., pheasants, emus). In Connecticut, outbreaks of EEE have occurred sporadically among horses and domestic pheasants since 1938, but no human cases have ever been confirmed. Human deaths, however, have occurred in nearby states.

In 1999, West Nile virus (WNV) was discovered in New York, New Jersey, and Connecticut. The WVN outbreak, in which seven humans and six horses died in New York and hundreds of birds died within the three states, was the first documentation of this virus in the Western Hemisphere. Unlike EEE, WNV is new to the Americas and native birds had not developed a natural immunity to this virus. Hence, a large

proportion of the birds bitten by WNV-infected mosquitoes died. Historically, sporadic outbreaks of WNV have occurred in parts of Africa and Eurasia since 1937. The virus is similar to the virus that causes St. Louis encephalitis (SLE) and causes similar symptoms in humans. Although WNV causes fatal illness in a smaller proportion of cases than EEE, its greater potential to cause large outbreaks makes it an important health concern.

3.3.4 Aesthetics

3.3.4a Scenery.Numerous studies have attempted to assign economic benefits to wetlands and open space, but quantifying the value of scenery for aesthetic purposes is extremely difficult. The draw and attachment that residents and visitors have for the Great Meadows Unit at Stewart B. McKinney NWR is largely due to the fact that the marsh is one of the only natural places in the area. The Refuge contributes to the aesthetic value of the area with its large tracts of wetlands, forest, and scrub-shrub. Refuge wetlands support sport and commercial fisheries, improve water and air quality, help control floods, support wildlife and provide outdoor recreation opportunities. In addition, people enjoy wetlands for their beauty, wildness and solitude.

3.3.4b Noise. Noise levels can be quite high at the Great Meadows Unit depending on proximity to roads and adjacent land uses e.g., Sikorsky Memorial Airport. US or State Highway 113 bisects the marsh and separates the Refuge portion from the airport, producing consistent background noise. Additionally, Interstate 95 is about 1 mile away from the Unit producing more background noise. Sikorsky Memorial Airport is comprised of 800 acres (324 ha) at an elevation of 9 feet (3 m) above mean sea level. It has two asphalt runways: 11/29 is 4,761 by 150 feet (1,451 x 46 m) and 6/24 is 4,677 by 150 feet (1,426 x 46 m). It has one asphalt helipad, 40 by 40 feet (12 x 12 m). In the year ending June 30, 2010 the airport had 67,951 aircraft operations, averaging 186 per day: 96% general aviation, 3% air taxi, and 1% military. Approximately 190 aircraft were then based at the airport: 74% single-engine, 13% jet, 11% multi-engine, and 3% helicopter. The Connecticut Wing Civil Air Patrol 022nd Stratford Eagles Composite Squadron (NER-CT-022) operates out of this airport (Airport IQ 5010 2017).

Chapter 4 ENVIRONMENTAL CONSEQUENCES This chapter provides an analysis of the effects of each of the alternatives on the physical, biological, and social environments discussed in Chapter 3. This analysis focuses on three aspects of each alternative – impacts associated with monitoring and surveillance activities, impacts associated with pesticide application, and impacts associated with wetland restoration projects. The impact analysis focuses on a programmatic-level approach to evaluate the effects of each alternative. The analysis of monitoring, surveillance, and pesticide use is presented at a project- specific level, while the analysis of wetland restoration projects is presented at a more general level because specific wetland restoration projects have not been developed at a site specific level. Further analyses of environmental consequences may occur when site-specific wetland restoration planning has been done. Where appropriate we have identified best management practices (BMPs) that should be implemented to avoid and minimize any potential environmental impacts.

The following resources would not be affected by any of the alternatives:

• Climate – None of the alternatives would change the climate in Connecticut. • Soils/Geology – None of the alternatives would have any effect on the soils or geology of the Refuge. • Environments – open water, mudflats, tidal marsh, and seasonal wetlands – None of the alternatives would change the abundance or distribution of the general environments found on the Refuge. While wetland restoration projects to improve tidal circulation may result in a different vegetative composition, they would not change the overall amount or location of environment. • Aesthetics – mosquito management activities are not expected to affect the scenery of the Refuge or noise levels at the Refuge. Wetland restoration projects to improve tidal circulation would minimally improve the aesthetic value of the Refuge. There would be no change to the noise environment. 42

• Noxious Weeds/Exotic Plants – Noxious weeds and exotic plants will continue to be part of the plant community on the Refuge. None of the alternatives are focused on eradicating noxious weeds or exotic plants. • Socioeconomics and Environmental Justice – All of the alternatives would occur within the boundary of the Refuge and do not involve the loss or acquisition of businesses, residential homes, or community facilities. Therefore, there are no adverse social or economic effects, and no minority or low-income populations or communities would be disproportionately affected under any of the alternatives.

These resources are not discussed further in this chapter.

In regard to pesticide effects measures, the following is a description of some of the measures used to assess effects of pesticides on plants and wildlife by the USEPA. These measures are used to describe potential pesticide effects to wildlife and plants as a result of mosquito management on the Refuge. “Measures of effect are obtained from a suite of registrant-submitted guideline studies conducted with a limited number of surrogate species and/or from acceptable open literature studies” (USEPA 2004, USFWS/NMFS 2004). The acute measures of effect routinely used for listed and non-listed animals in screening level assessments are the LD50, LC50, or EC50, depending on taxa. LD stands for "Lethal Dose", and LD50 is the amount of a material, given all at once, that is estimated to cause the death of 50% of a group of test organisms. LC stands for “Lethal Concentration” and LC50 is the concentration of a chemical that is estimated to kill 50% of a sample population. EC stands for “Effective Concentration” and the EC50 is the concentration of a chemical that is estimated to produce some measured effect in 50% of the test population. Endpoints for chronic measures of exposure for listed and non-listed animals are the NOAEL or NOAEC. NOAEL stands for “No Observed-Adverse-Effect-Level” and refers to the highest tested dose of a substance that has been reported to have no harmful (adverse) effects on a test population. The NOAEC (i.e., “No- Observed-Adverse-Effect-Concentration”) is the highest test concentration at which none of the observed results were statistically different from the control.”

4.1 Impacts on the Physical Environment

4.1.1 Impacts to Air Quality. 4.1.1a Alternative A – Proposed Action. Regular mosquito monitoring and surveillance activities would not have any adverse effects on air quality. Mosquito monitoring and surveillance activities are limited to checking carbon-dioxide baited light traps and Gravid traps at two locations within a mile of the Great Meadows Unit at the Refuge and dipping for larvae or pupae at the historic sites located on the Refuge. Monitoring and surveillance activities on the Refuge can occur by foot. This work is currently conducted by the CT MMP and the proposed action would not increase the number of monitoring trips needed, therefore, there would be no effect to air quality.

The aerial application of pesticides (adulticides only) where large areas of control are needed (e.g., greater than 50 acres) could result in aerial drift of pesticides. To minimize potential impacts, wind speed restrictions on spraying would be employed. Application of pesticide would be in compliance with the CT MMP WNV Surveillance and Response Plan.

Constructing wetland restoration projects with a focus of improving tidal circulation in order to reduce mosquito production could affect air quality by temporarily increasing vehicle related emissions. Depending on the size of the restoration project this temporary increase in emissions could range from one month to several months.

Temporary, localized dust may also occur as a result of restoration or enhancement projects. As site-specific projects are identified, potential air quality effects would be further analyzed. Best management practices to minimize any effects to air quality would be identified in a project-specific document.

4.1.1b Alternative B – Monitoring and Larvacides: Under this alternative no adulticides would be used to control mosquitoes. This would eliminate emissions associated with aircraft used to apply adulticides.

4.1.1c Alternative C - No Action. Under this alternative there would be no control of mosquitoes on the Refuge but monitoring would continue. The effects of this alternative on air quality are specific to emissions from the vehicles used for monitoring. The CT MMP would continue monitoring and surveillance activity at locations off of the Refuge. Under this alternative no pesticides would be used to control mosquitoes. This would eliminate emissions associated with aircraft used to apply pesticides. This alternative does include the construction of wetland restoration projects aimed at reducing mosquito production. As described under Alternative A, increases in vehicle related emissions and localized dust would be temporary in nature. As site-specific restoration projects are identified, potential air quality effects would be further analyzed.

4.1.2 Impacts to Topography. 4.1.2a Alternative A - Proposed Action. Regular mosquito monitoring and surveillance, and the application of pesticides for mosquito control proposed in this alternative would have no effect on topography. None of the monitoring and control work would result in any physical changes to the existing topography.

Under this alternative we would construct wetland restoration projects to improve tidal circulation and reduce the production of mosquitoes. Restoration projects are likely to change site topography. As restoration projects are developed we would analyze changes in topography in project-specific documents. These changes would be beneficial because they would result in increased tidal circulation that promotes native tidal marsh regeneration and a reduction in mosquito production.

4.1.2b Alternative B – Monitoring and Larvacides. Impacts under this Alternative would be the same as described under Alternative A.

4.1.2c Alternative C - No Action. This alternative does not include mosquito monitoring and control on the Refuge. Potential impacts from wetland restoration projects would be the same as described under Alternative A.

4.1.3 Impacts to Water 4.1.3a Alternative A - Proposed Action. Under this alternative, the regular mosquito monitoring and surveillance activities conducted by the CT MMP would continue. Monitoring and surveillance activities would not affect water resources because on the Refuge this work consists of walking to the sample sites. No contaminants are introduced to the Refuge’s water resources.

The application of pesticides on the Great Meadows Unit at the Refuge could affect water resources because pesticide application is occurring in an aquatic environment. We do not expect application of pesticides to result in any adverse effects to water quality on the Refuge. Bti and Bs are not expected to have any measurable effect on water quality. Bti and Bs are naturally occurring in most aquatic environments. There are no established standards, tolerances or USEPA approved tests for Bs. Surfactant (Agnique). This surfactant is considered “practically nontoxic” by the USEPA.

Pyrethrin/pyrethroid is extremely toxic to aquatic life, especially fish (Ecotoxnet 1994, USEPA 2006). Pyrethrum compounds are broken down in water to nontoxic products (Ecotoxnet 1994). Pyrethrin/pyrethroids are inactivated and decomposed by exposure to light and air. Pyrethrin/pyrethroids are also rapidly decomposed by mild acids and alkalis.

Best management practices for the application of pesticides would be listed in the SUP and include: Where mosquito control is needed based on established tresholds and survelillance data, use the most effective means that pose the lowest risk to abiotic and biotic resources. • Apply pesticides where monitoring and surveillance data justify its use. Employ wind speed restrictions on spraying. • The application of pyrethrin/pyrethroids will only occur in rare instances when a risk of serious human disease or death, or high risk to public health has been documented by a public health agency whose jurisdiction includes the Refuge. The application of pyrethrin/pyrethroids should occur at an ultra-low volume (according to pesticide label instructions and per habitat type). In a season when a risk of serious human disease or death, or high risk to public health is documented, the application of pyrethrin/pyrethroids must be limited to reduce impacts to habitat and wildlife, but sufficient to ensure effective mosquito control. Avoid application of pesticides during high tides and avoid open water areas of the Refuge (e.g., sloughs, channels, open bay).

Construction of wetland restoration projects is likely to change the amount and location of surface water on the Great Meadows Unit at the Refuge. Because these restoration projects would be constructed to improve tidal circulation, this change would be beneficial to water resources on the Refuge by decreasing anaerobic conditions and improving conditions for aquatic vertebrate and invertebrate communities. Implementing wetland restoration projects could also provide an increase of the floodwater capacity of the Refuge, which would reduce the risk of local flooding. Restoration projects would also have some short term, localized impacts to water quality from construction activities. Best management practices would be developed as these projects proceed to avoid or minimize impacts to water resources on the Refuge. The potential effects of restoration projects on water resources of the Refuge would be analyzed in project-specific documents.

4.1.3b Alternative B – Monitoring and Larvacides. The impacts of this alternative would be the same as described under Alternative A as applicable to the use of Bti and Bs only.

4.1.3c Alternative C - No Action. Under this alternative no mosquito monitoring, surveillance, or control would be allowed on the Refuge. Therefore there would be no effect to water resources on the Refuge from either monitoring or control. Potential effects from restoration projects would be the same as Alternative A.

4.2 Impacts on Biological Resources

Table 6 summarizes the biological impacts of pesticides that would be used under Alternatives A and B. No pesticides will be used in Alternative C.

Table 6. Summary of biological impacts of the pesticides used for mosquito control in wetlands by the CT MMP.

Name Degradation Effects on Wildlife Effects on humans Bacillus thuringiensis Foliar HL 1-4d, soil HL Nontoxic to birds and fish, Adverse impacts not 1, 2, 3, 4, 5 israelensis (Bti) mean 4mo. minimal toxicity to bees and likely from label other non-target insects; applications of Bti on the indirect food web effects may Refuge. occur, especially among higher order vertebrates (e.g., birds). Bacillus sphaericus TBD – cannot find in Various tests revealed no Adverse impacts not (Bs) 2362 serotype literature expected harm to non-target likely from label H5a5b, strain ABTS organisms. (EPA - applications of Bs on the 17439 https://www3.epa.gov/pestici Refuge. Avoid contact des/chem_search/reg_actions/ with granules. registration/fs_PC- 119803_06-May-14.pdf) 8 Monomolecular film Environmental fate not Films are potentially lethal to May cause eye irritation; determined any aquatic insect that lives otherwise acute and chronic on the water surface or effects are unknown. requires periodic contact with the air-water interface to obtain oxygen; other ecotoxicological effects are

unknown (e.g., birds, fish). 6, 7, 9 Pyrethrin/Pyrethroids Soil HL 12d, rapidly Toxic to aquatic life Slight acute oral toxicity to degrades in sunlight and including fish, insects, and mammals, allergic reaction water. aquatic invertebrates, slightly possible.

toxic to vertebrates.

1Data taken from National Pesticide Information Center for Bt (2015), 2Poulin et al. 2010, 3Siegel and Shadduck 1992 (for Bti), 4Appendix I (2003), 5Appendix H, 6National Pesticide Information Center (1998), 7Extension Toxicology Network (1994), 8Agnique Material Safety Data Sheet (2006), USEPA 2006. 9 US Environmental Protection Agency fact sheet on Bacillus sphaericus

4.2.1 Impacts to Vegetation 4.2.1a Alternative A - Proposed Action. Impacts to vegetation could occur during access (on-foot) within tidal marsh to conduct mosquito management. There are three state listed plant species of concern that occur at the Great Meadows site: salt marsh pink (endangered), beach needlegrass (endangered), and bayonet grass (species of concern). Walking in the tidal marsh can also introduce and spread invasive weeds. The following best management practices – which would be included in the SUP - would be implemented under the Proposed Action to reduce negative impacts to vegetation: • Use existing pathways or limit the number of travel pathways used by people on foot within the marsh. • Designate pathways for CT MMP staff – to ensure no trampling of state-listed species. • Apply pesticide (larvicide or pupacide) by foot – no vehicles would be allowed in the marsh.

• Refuge staff and the CT MMP would develop and implement measures to protect plant resources of concern and to reduce the introduction and spread of invasive weeds by mosquito management activities during all mosquito control operations. Use the most specific - least toxic to non-target organisms - possible. The use of broad spectrum pesticides will only be used under emergency situations in discrete locations. • Invasive weeds would be controlled through manual removal and chemical control.

The application of pesticides, including adulticides, are not likely to adversely affect vegetation directly because the pesticides used for mosquito control are not known to harm plants (Table 5). How reductions in certain invertebrate populations, as a result of repeated pesticide applications, would impact specific invertebrate-plant interactions (e.g., pollination) within tidal marsh of the Refuge is not known.

Best management practices would also include controlling invasive weeds, both manually and through chemical control where possible. The Refuge staff and CT MMP would meet annually to discuss how to reduce the spread of invasive weeds from mosquito management activities.

Tidal circulation is a critical component in restoring native vegetation. Areas of the Refuge submerged in persistent shallow water typically have poor vegetative health relative to other tidal areas (i.e., reduced species richness, cover, and height). This condition is exacerbated by mosquito management activities described above. However, projects designed to restore or enhance tidal circulation and subsequently reduce mosquito production, would also improve habitat conditions for native vegetation. Restoring the hydrology would also have the added benefit of reducing any direct impacts to the vegetation from accessing the marsh for mosquito control activities. Wetland restoration projects may have some temporary, negative, short-term impacts to vegetation during project implementation (e.g., use of construction equipment in the marsh). A detailed evaluation of vegetation impacts from restoration projects would be completed as site-specific projects are identified.

4.2.1b Alternative B – Monitoring and Larvacides. Same as described under Alternative A.

4.2.1c Alternative C - No Action. Under this alternative mosquito monitoring would take place. Therefore, access to the sampling sites may contribute to vegetation trampling (see Alternative A). However, there would be less trampling under this alternative since no mosquito control activities would take place on the Refuge. The effects of wetland restoration projects are the same as described under Alternative A.

4.2.2 Impacts on Mammals 4.2.2a Alternative A - Proposed Action. The Proposed Action has the potential to impact common mammals through the application of pesticides and the disturbance of having humans present at the site. However, a request to the National Diversity Database in March 2017 revealed that the Great Meadows site was not identified as a potential location for any mammal species of concern.

Adverse impacts to mammals may occur as a result of marsh access via foot for mosquito management activities. Under the Proposed Action we would implement the following best management practices – which would be included in the SUP - to reduce potential impacts: • Use existing pathways or limit the number of travel pathways used by mosquito control staff within the marsh. • Refuge staff and the CT MMP would develop and implement measures to reduce disturbance to mammals and to protect resources of concern during all mosquito control operations.

The use of larvicides and pupacides for the purpose of mosquito management are not likely to directly affect native mammal populations of the Refuge. Larvicides and pupacides allowed under this alternative include Bti, Bs, and monomolecular films (Agnique). New products will not be applied without prior Refuge approval. The CT MMP and Refuge will communicate regarding existing and new mosquito larvicides and 47

adulticides that minimize non-target effects. Adverse effects on mammals from Bti, Bs, and Agnique (monomolecular film) are not expected (Appendix H, I) when applied according to the label instructions.

Furthermore, label rate application of pesticides is far below LD and LC50 toxicity data (pers. comm., R. Wolfe). Bti and Bs have practically no acute or chronic toxicity to mammals, birds, fish, or vascular plants (USEPA 1998, USEPA 2014). Extensive acute toxicity studies indicated that Bti and Bs are virtually innocuous to mammals (Siegel and Shadduck 1992 and USEPA Bs factsheet). Studies exposed a variety of mammalian species to Bti and Bs at moderate to high doses and no pathological symptoms, disease, or mortality were observed.

Under the Proposed Action we would also allow the use of pyrethrin/pyrethroid pesticides on the Refuge under certain conditions. Oral exposure of pyrethrin/pyrethroids could occur through consumption of plants or plant parts that have been sprayed (ground-based application). A terrestrial exposure model showed no acute or chronic risks to mammal or bird species (USEPA 2006). To reduce any potential negative effects on mammals the Refuge would require the following best management practices – which would be included in the SUP - for the CT MMP when applying adulticides: • The application of pyrethrin/pyrethroids occurs in rare instances when a mosquito-borne disease incidence has been documented on the Great Meadows Unit at the Refuge or within flight range of vector mosquito species present on the Refuge; adult vector mosquito thresholds decided by the CT MMP are exceeded on the Refuge; and when there are no practical and effective alternatives to reduce a mosquito-borne, disease-based health threat. • The application of pyrethrin/pyrethroids must occur at an ultra-low volume (according to pesticide label instructions and per habitat type). • The application of pyrethrin/pyrethroids must occur where monitoring and surveillance data justify its use. • In a season when a risk of serious human disease or death, or high risk to public health is documented, the application of pyrethrin/pyrethroids must be limited to reduce impacts to habitat and wildlife, but sufficient to ensure effective mosquito control.

Wetland restoration projects that improve tidal circulation and reduce shallow ponding are likely to improve mammalian habitats by improving vegetation health and reduced need for mosquito management and associated disturbance. Significant adverse effects to mammalian species are not likely to occur during restoration or enhancement projects because areas with poor hydrology typically exhibit low habitat quality for mammals, especially small mammals. Best management practices to reduce effects would be developed and implemented to reduce direct effects during project construction. As restoration projects progress, additional site-specific analysis will be completed and methods to avoid and minimize any potential direct effects to mammals will be identified.

4.2.2b Alternative B – Monitoring and Larvacides. Under this alternative, impacts related to access for mosquito management and impacts related to wetland restoration or enhancement projects are the same as described under Alternative A. This alternative would include the application of Bti and Bs for mosquito control. Both are not known to directly affect vertebrate species (Appendix H, I).

4.2.2c Alternative C - No Action. This alternative includes mosquito monitoring and surveillance. Therefore there would be some effects to mammals from mosquito management activities under this alternative (see Alternative A site access). This alternative does include wetland restoration projects and effects to mammals from these activities would be the same as described in Alternative A.

4.2.3 Impacts on Birds 4.2.3a Alternative A - Proposed Action. Impact to birds that use the Great Meadows Unit at the Refuge may occur during Refuge access for mosquito monitoring, surveillance and control, as well as the application of pesticides. The birds of concern to the Refuge in the salt marsh are the seaside sparrow (state threatened), 48

saltmarsh sparrow (state species of concern), and clapper rail (declining populations in CT). The National Diversity Database request identified seven additional species that could occur on the Refuge. None of these species nest at the site; they may only be present during migration. These species are the short-eared owl, least bittern, American bittern, king rail, common moorhen, and pied-billed grebe.

Impacts to birds may occur; birds may be temporarily flushed and nests may be trampled as a result of ground access via foot. A plan would be created by the Refuge in conjunction with the CT MMP to identify access routes to the mosquito management areas so that disturbance to these species will be minimized. Birds will be able to return to roosting and nesting sites once operations have ceased in the area. The Refuge staff has data from 2009 documenting where salt marsh and seaside sparrows nested. In these areas, and any future known areas, dip site would be established to avoid harming these species and their nests. These would be the only mosquito dip sites in these areas throughout the annual surveillance and monitoring. If the CT MMP needs to spray in these areas, Refuge staff would survey the area for sparrow nests (as well as rail) and inform CT MMP staff of any nest to avoid trampling.

The use of pesticides for the purpose of mosquito management may directly or indirectly affect resident and migratory bird populations on the Refuge. Pesticides allowed under this alternative include Bti and Bs, monomolecular films (Agnique) and pyrethrin/pyrethroids.

Bti and Bs have practically no acute or chronic toxicity to mammals, birds, fish, or vascular plants (USEPA 2014 and USEPA Bs factsheet; Appendix H, I). The USEPA states that various tests were conducted for Bs, and they revealed no expected harm to non-target organisms. There is the potential for Bti to kill midge larvae (family chironomidae). Chironomid (non- biting midge) larvae can be abundant in wetlands and form a significant portion of the food base for other wildlife, including birds (Batzer et al. 1993; Cooper and Anderson 1996; Cox et al. 1998). As with Bti, there is concern regarding potential negative impacts to chironomid larvae from methoprene. Some studies have suggested methoprene impacts to other organisms that may form part of the food base for birds. McKenney and Celestial (1996) noted significant reductions in the number of young produced in mysid shrimp at 2 ppb. Sub-lethal effects on the cladoceran, Daphnia magna, in the form of reduced fecundity, increased time to first brood, and reduced molt frequency have also been observed at lower concentrations of methoprene (Olmstead and LeBlanc 2001).

Surface oils may adversely affect wildlife, such as oiling the feathers of young waterfowl and other birds. This may be of particular concern at low temperatures when the oil and lack of feather function could affect thermoregulation (Lawler et al. 1998). Pyrethrin/pyrethroid pesticides would be the only adulticide chemicals permitted for use on the Refuge. Pyrethrin/pyrethroids are not considered toxic to birds (Milam et al. 2000, USEPA 2006) when applied at labeled rates. See Appendices I and K for non-target effects of pyrethrin/pyrethroids on vertebrates and invertebrates.

However, non-target effects to birds from pesticide application may occur as a result of reduced food base (e.g., Chironomid invertebrates). For example, monomolecular films are potentially lethal to any aquatic insect that lives on the water surface or requires periodic contact with the air-water interface to obtain oxygen (Appendix H, I). There is uncertainty with regard to pyrethrin/pyrethroids, which have been shown to have no impact on large-bodied arthropods, but have been shown to reduce invertebrate populations, especially among small-bodied arthropods (Boyce et al. 2007). Characteristics of areas that require mosquito control in the Great Meadows marsh include poor tidal hydrology and dense monocultures of invasive species e.g., Phragmites. The habitat quality in these areas is poor for a variety of wildlife species. Therefore, potential indirect impacts to the food chain as a result of pesticide application within these areas are likely to be limited because of existing low quality habitat conditions for many estuarine-dependent species.

The reduction of mosquito production through improvement of tidal circulation and wetland quality would benefit both resident and migratory bird populations. Benefits would include improved water quality

(reduced pesticide application), reduced human and vehicular disturbance, and improved primary productivity and food resources (e.g., invertebrates, fish). Short-term negative impacts to birds during restoration or enhancement projects (aimed at reducing mosquito production) may occur as a result of construction activities. Techniques to reduce effects (e.g., exclusionary fencing, timing, equipment types) would be used to reduce impacts during project construction. As restoration projects are identified, a project specific document would be prepared that addresses any project- specific potential impacts.

Significant mosquito production and absence of control may negatively impact bird populations. Although mosquitoes themselves are a part of estuarine ecosystems, they are known vectors of disease, including diseases that cause harm to humans and wildlife (e.g., West Nile Virus). Mosquito-borne diseases such as West Nile Virus (WNV) have shown to be lethal to wildlife. As of 2011, 326 bird species have been listed in the Center for Disease Control WNV avian mortality database (http://www.cdc.gov/ncidod/dvbid/westnile/birdspecies.htm, accessed May 2, 2011). The list includes wildlife that inhabit tidal marsh such as waterfowl, grebes, heron, egrets, cormorants, songbirds (wrens, yellowthroats, song sparrows), and rails (clapper rail). Other vertebrates known to be infected by WNV include horses, bats, chipmunks, skunks, rabbits, and squirrels.

Research conducted in the eastern U.S. in 2002 when the WNV outbreak was in full swing, found fewer incidences of WNV in humans in areas with a diverse array of bird species (Swaddle and Calos 2008). This link between higher bird diversity and reduced human WNV infection is attributed to the fact that crows, jays, thrushes, and sparrows are competent (amplifying) hosts of the WNV, making them able to contract the disease and pass it on through a vector more efficiently. When bird diversity is low, competent host species tend to represent a higher proportion of the bird population, increasing the likelihood that a mosquito will encounter an infected bird and transmit the virus during its next bite. A diverse suite of bird species, with large numbers of incompetent hosts in the population, reduces the transmission rate to other birds or mammals, including humans. A similar study showed increased mammalian diversity decreased Lyme disease risk to humans (LoGiudice et al. 2003).

To reduce the potential for negative impacts to bird populations the following best management practice would be implemented under this alternative and included in the SUP: • Access (via foot) to tidal marsh for the purpose of mosquito management would be seasonally limited (e.g., reduced access during high tide events) from May 15 to September 1 in areas that are known to be occupied by marsh species of concern e.g., clapper rail, saltmarsh sparrow, and seaside sparrow. • In sites known or historically known to have nesting species of concern, dip sites would be established by Refuge staff in conjunction with CT MMP at the annual meeting. These would be the only dip sites that can be used in these areas during monitoring/surveillance for the year. • If spraying needs to occur in nesting sites, Refuge staff would survey those areas for nests and inform CT MMP staff of nest locations to avoid trampling. • Provide CT MMP with training on measures to avoid impacts to wildlife. • Application of pyrethrin/pyrethroids would be informed by monitoring of mosquito vector populations and surveillance indicating location of disease prevalence. • Application of pyrethrin/pyrethroids would be a limited to reduce impacts to wildlife and habitat but sufficient to prevent a second adult emergence. • When applying pesticides, the pesticide would be applied according to pesticide label instructions and per habitat type. Appropriate training for pesticide applicators would occur to ensure pesticide labels are followed.

The potential also exists for the transmission of mosquito-borne diseases, which have been shown to cause mortality in birds (e.g., WNV). Impacts to birds as a result of physical access (trampling of vegetation, nests) and application of pesticides (food chain effects) as a result of mosquito management could occur, but

it is unlikely that these actions would significantly affect bird populations on the Refuge as long as access to the mosquito management sites is planned and implemented according to species occurrence.

4.2.3b Alternative B – Monitoring and Larvacides. See Alternative A for effects related to access and wetland restoration or enhancement projects. Bti and Bs are considered for mosquito control under this alternative. Neither is known to directly affect vertebrate species although indirect impacts may occur as a result of reduced invertebrate populations (See Appendix H, I).

If mosquitoes are not adequately controlled at the larval stage, adult populations will develop and short-term control measures will not be available (e.g., adulticides). Significant adult mosquito production may produce negative effects on some bird populations. Although mosquitoes themselves are a part of estuarine ecosystems they are known vectors of disease, including diseases that cause harm to humans and wildlife (e.g., WNV). Mosquito-borne diseases such as WNV have been shown to be lethal to wildlife.

This alternative would also include wetland restoration projects. The effects of wetland restoration projects are described under Alternative A.

4.2.3c Alternative C –No Action. Under this alternative, there is a monitoring component. Just as in Alternative A, CT MMP must follow the BMPs for reducing the disturbance to birds and avoiding trampling of nests. See Alternative A for impacts related to wetland restoration or enhancement projects.

4.2.4 Impacts on Reptiles and Amphibians 4.2.4a Alternative A - Proposed Action. Under this alternative the CT MMP would conduct mosquito monitoring and surveillance, mosquito control through application of pesticides and tidal wetland restoration. The National Diversity Database request from March 2017 listed northern diamondback terrapins as the only reptile species of concern. No amphibians were listed.

Pesticide effects on reptiles and amphibians may occur through reductions in insects that serve as food source (Hoffman et al. 2008), through direct individual effects from pesticide application, or from trampling of individuals or habitat. Birds are often used as surrogates to estimate or predict the effects of pesticides on reptiles whereas fish are often the surrogates for amphibians (Hoffman et al. 2008). Direct chronic effects have been found for the San Francisco garter snake from application of labeled rates of permethrin (synthetic pyrethroid, Hoffman et al. 2008). This species does not occur anywhere near the Refuge (and is in fact a western species) but these findings suggest other reptiles may incur direct chronic effects. Best management practices to reduce potential adverse effects of pyrethrin/pyrethroid would be included in the SUP and include: The application of pyrethrin/pyrethroids would only occur in rare instances when a risk of serious human disease or death, or high risk to public health has been documented by a public health agency whose jurisdiction includes the Refuge. The application of pyrethrin/pyrethroids should occur at an ultra-low volume (according to pesticide label instructions and per habitat type). The application of pyrethrin/pyrethroids should occur where monitoring and surveillance data justify its use (e.g., incidence of mosquito-borne disease, exceedance of tolerance limits for adult mosquitoes). In a season when a risk of serious human disease or death, or high risk to public health is documented, the application of pyrethrin/pyrethroids must be limited to reduce impacts to habitat and wildlife, but sufficient to ensure effective mosquito control.

4.2.4b Alternative B – Monitoring and Larvacides. The impacts to reptiles and amphibians of this alternative are the same as those described under Alternative A. This alternative would only use Bti and Bs which are not known to affect reptiles and amphibians – although some food web changes could occur (see Alternative A). 51

4.2.4c Alternative C - No Action. Under this alternative the CT MMP would conduct mosquito monitoring. This alternative also includes wetland restoration projects. See Alternative A for a description of potential impacts from access and wetland restoration projects.

4.2.5 Impacts on Fisheries 4.2.5a Alternative A – Proposed Action. Mosquito monitoring and surveillance activities are not expected to adversely affect fish because these activities do not occur within open sub tidal waters of the Great Meadows Unit at the Refuge (e.g., sloughs, channels, open bay) and are not expected to adversely affect water quality (e.g., turbidity, dissolved oxygen). The Proposed Action does include the application of larvicides, pupacides and adulticides under certain conditions.

Direct effects on fish populations are not expected from proposed larvicides and pupacides (Appendices H, I). However, the application of adulticides has the potential to adversely affect fish populations (Gunasekara 2005). Pyrethroids are considered highly toxic to fish and invertebrates (Appendices I and K). Fish sensitivity to the Pyrethrin/Pyrethroids may be explained by their relatively slow metabolism and elimination of these compounds.

Pesticide application may result in indirect impacts to fish species by reducing their invertebrate prey base. Adverse impacts to invertebrates are expected from pesticide application. However, the application of pesticides on discrete areas of the Refuge is meant to lower mosquito numbers, but not eliminate mosquito populations entirely (or other invertebrate species), ensuring food availability for fish. Also, application of pesticides would occur in site specific locations and would not be applied indiscriminately across the entire Refuge.

To avoid adverse impacts to fish species the following best management practices – also outlined in the SUP - would be used: Application would be restricted to those specific areas where a pathogen is present and mosquito population thresholds have been exceeded on the Great Meadows Unit at the Refuge, and where application can be effectively treated while minimizing non-target effects, especially to endangered species. If beneficial, the CT MMP should conduct simultaneous application of larvicides with the adulticide application to prevent future adult outbreaks When applying pesticides, the pesticide would be applied according to pesticide label instructions and per habitat type. Apply pesticides only to discrete, mosquito producing areas of the Refuge. Appropriate training for pesticide applicators would occur to ensure pesticide labels are followed.

This alternative also includes construction of wetland restoration projects to improve tidal circulation. Areas of the Great Meadows Unit at the Refuge where above average mosquito production occurs also contain swales where water impounds following high tides. These swales do not drain into tidal channels and can entrap fish that forage within tidal marsh during higher tides. We expect that wetland restoration projects aimed at reducing mosquito production will reduce effects related to entrapment and will improve foraging and nursery habitat. Reduced mosquito production would also reduce the need for pesticide application which would also benefit fisheries. As restoration projects are developed, additional project-specific analysis will be completed on the short-term impacts and long- term benefits to fish.

The frequency of conditions that would require use of adulticides on the Refuge has been rare over the past few decades. No requests have been made in the past even with the presence of West Nile virus in the region.

This pattern suggests that future use of adulticides in discrete areas of the Refuge is unlikely, but if occurred, the frequency and scope of application is not likely to cause significant adverse impacts to fish and invertebrate populations. Applications would likely occur along the upland edge where adult mosquitoes coming off the Refuge’s wetlands rest before flying into neighboring locales. Applications would not be made over water, as adult mosquitoes would unlikely be located there. Therefore, potential impacts to fish would be negligible.

4.2.5b Alternative B – Monitoring and Larvacides. Mosquito monitoring and surveillance activities would have no effect on fisheries because none of this work takes place in the water. No adverse effects on fish are expected from the application of approved larvacides for mosquito control. Wetland restoration projects are part of this alternative and potential impacts are described in Alternative A.

4.2.5c Alternative C - No Action. Mosquito monitoring would be permitted under this alternative; therefore, there would be no impacts to fish. Wetland restoration projects are part of this alternative and potential impacts are described in Alternative A.

4.2.6 Impacts on Invertebrates 4.2.6a Alternative A – Proposed Action. Monitoring and surveillance activities are not expected to adversely impact invertebrate populations. Chemical treatment of mosquito populations on the Great Meadows Unit at the Refuge has the potential to adversely impact invertebrates and these are described below.

Bacillus thuringiensis israelensis (Larvicide) – The impact on local populations of invertebrate species over time with periodic and continued use of Bti is unknown, but potential for negative effects is a possibility (Appendices H, I). Host range and effect on non-target organisms indicates that Bti is relatively specific to the Nematocera suborder of Deptera, in particular filter-feeding mosquitoes (Culicidae) and blackflies (Simuliidae) (Glare and O’Callaghan 1998). Bti is pathogenic to some species of midges (Chironomidae) and Tipulidae, although to a lesser extent than mosquitoes and biting flies and is not reported to affect a large number of other invertebrate species (Glare and O’Callaghan 1998). Bti concentration is important with regard to effects on non-target organisms. Of particular concern is the potential for Bti to kill midge larvae (family Chironomidae). Chironomid (non-biting midge) larvae are often the most abundant aquatic insect in wetland environments and form a significant portion of the food base for other wildlife (Batzer et al. 1993; Cooper and Anderson 1996; Cox et al. 1998). Reduced invertebrate populations as a result of food web effects (e.g., reduction of nematoceran Diptera) have been shown in studies of Bti (Hershey et al. 1998).

Bacillus sphaericus (Larvicide) – Field and laboratory studies of the effects of Bs have found that its toxin is more precise then that of Bti’s and has been determined safe for a large variety of non-target invertebrates, including a number of mosquito predators (Toxorhynchites), non-biting midges (Chironomidae) and other species of long horned flies (Nematocera; Lacey 2007). High concentration of Bs can adversely affect some genera of Toxorhynchites (e.g., tree hole mosquito) when the species is in its first instar (Lacey 2007) but does not affect the elephant mosquito (Toxorhynchites rutilus), which is found in Connecticut. A three year study conducted in southeastern Wisconsin by Merritt et al. (2005) assessed the ecological effects of Bs when used for mosquito control and found that there were no detrimental effects on non-target organisms. Few long-term effects of repeated treatment with Bs on aquatic community structure and diversity have been reported (Mulla et al. 1984; Lacey and Mulla 1990).

Monomolecular film (Agnique) – Monomolecular films are potentially lethal to any aquatic insect that lives on the water surface and requires periodic contact with the air-water interface to obtain oxygen (USFWS 2004). The film interferes with larval orientation at the air-water interface and/or increases wetting tracheal structures, thus suffocating the organism. As the film spreads over the water surface, the treatment tends to concentrate the larvae, which may increase mortality from crowding stress (Dale and 53

Hulsman 1990). Surface oil is effective against all immature stages through suffocation; but studies have demonstrated negative effects on water surface-dwelling insects from applications of oils (Mulla and Darwazeh 1981; Lawler et al. 1998).

Pyrethrin/pyrethroid (adulticide) – Under the Proposed Action, pyrethrin/pyrethroid pesticides could be applied when there is high risk of mosquito-borne disease (Section 2.2.1, Phase 5). Pyrethrin/pyrethroids are known to cause acute toxicological impacts to benthic invertebrates at rates used for mosquito abatement (USEPA 2006). Because pyrethrin/pyrethroids are broad-spectrum insecticides, they are potentially lethal to most insects, including both terrestrial and benthic forms. All adulticides are very highly toxic to aquatic invertebrates in low concentrations (e.g., 1 ppb) (Milam et al. 2000). Because most adulticides can be applied over or near water when used for mosquito control, there are risks to aquatic invertebrates from direct deposition and runoff of the pesticides. These chemicals may affect the saltmarsh tiger beetle, the only invertebrate species of concern that may occur on the site.

In order to reduce adverse impacts to invertebrate populations the following best management practices (also listed in the SUP) would be required. • The CT MMP would be required to minimize the use of pesticides and continually investigate formulations and compounds that are least damaging to fish and wildlife populations. • The CT MMP would be required to review the past year’s pesticide proposals and submit any changes in the pesticides or formulations of pesticides that they expect to use in the upcoming year. This information should be made available at or before the time of the annual meeting. • Pesticide would be applied according to pesticide label instructions and per habitat type. Apply pesticides only to discrete, mosquito producing areas of the Refuge. Appropriate training for pesticide applicators would occur to ensure pesticide labels are followed. Application of pesticides would be informed by monitoring mosquito vector populations and surveillance indicating location of disease prevalence. • The CT MMP would adapt methods to reduce ecological risk to the environment (e.g., boom height, droplet size, application rate) as new information on ecological risk and avoidance measures are identified by appropriate regulatory agencies. • The CT MMP would follow all the best management practices for pesticide application stated in this document including ultra-low volume application and limiting the amount of pesticides applied.

Under this alternative, we would also plan and implement wetland restoration or enhancement projects to reintroduce or improve tidal circulation. Improved tidal circulation is likely to improve invertebrate communities while reducing above-average mosquito production. Where tidal marsh restoration or enhancement has occurred on the Refuge, the need for mosquito management has been significantly reduced or eliminated.

Studies of invertebrate populations in tidal marsh with poor hydrology (e.g., shallow water impoundments) that require mosquito management have not been conducted. Available data on wildlife suggest these areas of the Refuge provide poor habitat for a variety of species. Poor habitat conditions could also exist for invertebrate communities, leading to depressed or absent higher order predators (e.g., birds, small mammals). Although some studies have shown direct and indirect effects of pesticides, little is unknown which of two conditions, poor hydrological conditions or repeated pesticide applications, would contribute to depressed invertebrate populations.

4.2.6b Alternative B – Monitoring and Larvacides. There would be no adverse impacts to invertebrates from continued mosquito monitoring and surveillance activities. Under this alternative we would allow the application of Bti and Bs, which are non-chemical pesticides. Use of Bti may have a negative effect on invertebrate population as described under Alternative A. This alternative also includes the construction of wetland restoration projects aimed at restoring tidal circulation. Restoring tidal circulation would reduce the

need to apply non-chemical pesticides and may have a positive effect on vertebrate populations, as described in Alternative A.

4.2.6c Alternative C - No Action. Under this alternative no effects to invertebrates are expected because no pesticides would be applied at the Refuge. Restoration of tidal marsh to reduce mosquito production is likely to benefit other invertebrate populations (see Alternative A).

4.3 Impacts on the Social Environment

4.3.1 Impacts on Cultural and Historic Resources 4.3.1a Alternative A – Proposed Action. Mosquito monitoring and surveillance activities are not likely to have any adverse effects to cultural resources because these activities are limited to walking on the Great Meadows Unit at the Refuge and sampling various mosquito production areas. The application of pesticides is also unlikely to have any adverse effect to cultural resources because they target mosquito populations and are applied through targeted spraying.

Wetland restoration projects identified to restore tidal circulation could affect cultural resources. As individual restoration projects are identified, the Service will exercise Section 106 of the National Historic Preservation Act including consultation with the State Historic Preservation Officer (SHPO), in accordance with the programmatic agreement between the SHPO and the Service.

4.3.1b Alternative B – Monitoring and Larvacides. See Alternative A.

4.3.1c Alternative C - No Action. See Alternative A.

4.3.2 Impacts on Land Use 4.3.2a Alternative A – Proposed Action. Under this alternative, land use or access to the Great Meadows Unit at the Refuge may be limited depending on the activity. The use of adulticides or any qualified construction activity for marsh restoration that would put visitors at risk would be cause to temporarily close the trails and perhaps hunting activities at the Great Meadows Unit. These closures would be temporary and the activities would be permitted again when it was safe.

4.3.2b Alternative B – Monitoring and Larvacides. See Alternative A for construction temporary closures.

4.3.2c Alternative C - No Action. See Alternative A for construction temporary closures.

4.3.3 Impacts on Human Health and Safety Concerns 4.3.3a Alternative A –Proposed Action. The primary purpose of mosquito control on the Great Meadows Unit at the Refuge is to protect human health and safety. There would be no adverse impacts on human health and safety from mosquito monitoring and surveillance. Under this alternative, the potential exists for human exposure (i.e., CT MMP staff, Refuge visitors, and Refuge staff) to pesticides including adulticides. The need to adjust mosquito control techniques and monitor mosquito populations is expected to progress according to future influxes of mosquito-borne disease and mosquito breeding sites. To minimize the potential impacts of human exposure to pesticides on the Refuge, the following best management practices – also listed in the SUP - would be implemented: Prior to the application of any pesticide, the CT MMP must identify the treatment locations, treatment schedule, and identify the pesticide to be used. The CT MMP must provide this information to the Refuge Manager at least 48 hours prior to proposed application. The Refuge Manager would post a notice at the Refuge Office and at all visitor parking areas with information on the dates of adulticide application.

During application of adulticides, the Refuge would be closed to all visitors and Refuge staff in areas where application will occur.

Construction of wetland restoration projects to improve tidal circulation would have no adverse impacts to human health and safety. During construction, public access would not be allowed in the project area. Improving tidal circulation should reduce mosquito populations and improve human health and safety.

4.3.3b Alternative B – Monitoring and Larvacides. While the control of mosquito larvae and reduction in mosquito breeding sites should reduce the potential for disease outbreaks, the inability to use chemical larvicides and adulticides in the event of a disease outbreak could have an impact on human health and safety. The use of biological/biorational controls and habitat manipulation is designed to prevent the establishment of disease- carrying adult mosquito populations; however, tide and topographic features could combine to create unanticipated conditions that allow the emergence of adult mosquitoes at population levels higher than average. There would be no adverse impacts to human health and safety from mosquito monitoring and surveillance activities.

4.3.3c Alternative C - No Action. The lack of mosquito control may negatively affect human health and safety of Refuge visitors or surrounding communities. Seasonal wetlands, tidal marsh and dredge pond areas that do not have full tidal exchange may harbor disease vectors that spread to adjacent urban areas.

4.4 Cumulative Impacts

4.4.1 Alternative A – Proposed Action This action is likely to have cumulative impacts due to the long-term use of pesticides. Persistent use of access points reduces vegetation health along access routes. Persistent use of pesticides may depress invertebrate populations and alter food webs. The ultimate land management practice to reduce cumulative impacts of mosquito management is to enhance or restore tidal hydrology. These types of actions would reduce mosquito production to “natural” levels and promote a healthy tidal marsh ecosystem. Other best management practices – also included in the SUP - to reduce cumulative impacts include: • Limit the number of travel pathways used on foot within wetlands • To the extent feasible, minimize the use of pesticides and continually investigate formulations and compounds that are least damaging to fish and wildlife populations. • Apply pesticides according to pesticide label instructions and per habitat type. Apply pesticides only to discrete, mosquito producing areas of the Refuge. Appropriate training for pesticide applicators would occur to ensure pesticide labels are followed. • Refuge staff and the CT MMP will develop and implement measures to protect resources of concern and to reduce the introduction and spread of invasive weeds by mosquito management activities. Use the most specific - least toxic to non-target organisms - possible. The use of broad spectrum pesticides will only be used under emergency situations in discrete locations. • Invasive weeds will be controlled through manual removal and chemical control.

4.4.2 Alternative B – Monitoring and Larvacides. See Alternative A.

4.4.3 Alternative C - No Action. The cessation of mosquito control on the Great Meadows Unit at the Refuge may result in an increase the level of mosquito management in areas adjacent to the Refuge.

Chapter 5 COMPLIANCE, CONSULTATION, AND COORDINATION WITH OTHERS

5.1 Agency Coordination and Public Involvement The Plan and EA was prepared with the involvement of local partners including the Connecticut Mosquito Management Program, which includes the following state agencies: • Department of Energy & Environmental Protection (DEEP) • Connecticut Agricultural Experiment Station (CAES) • Department of Public Health (DPH) • Department of Agriculture and the Department of Pathobiology at the University of Connecticut (UCONN)

5.2 Environmental Review and Consultation As a federal agency, the Service must comply with provisions of NEPA. An EA was developed to evaluate reasonable alternatives that would meet stated goals and assess the possible environmental, social, and economic impacts on the human environment. This EA serves as the basis for determining whether implementation of the preferred alternative would result in a federal action significantly affecting the quality of the environment. The EA also acts as a vehicle for consultation with other government agencies and interface with the public in the decision- making process.

5.3 Other Federal Laws, Regulations, and Executive Orders In undertaking the preferred alternative, the Service would comply with the following federal laws, Executive Orders (EOs), and legislative acts: Intergovernmental Review of Federal Programs (EO 12372); Archaeological Resources Protection Act of 1979, as amended; Fish and Wildlife Act of 1956, as amended; Fish and Wildlife Conservation Act of 1980, as amended (16 USC 661-667e); Fish and Wildlife Improvement Act of 1978, aas amended; Endangered Species Act of 1973, as amended (16 USC 1531 et seq.); National Environmental Policy Act of 1969, as amended; Federal Noxious Weed Act of 1990, as amended; Floodplain Management, as amended (EO 11988); Protection of Wetlands (11990); National Historic Preservation Act of 1966, as amended; National Wildlife Refuge System Improvement Act of 1997, as amended; Antiquities Act of 1906, as amended; Protection and Enhancement of the Cultural Environment (EO 11593); Archaeological and Historic Preservation Act of 1974, as amended (PL 93-291; 88 STAT 174; 16 USC 469); Environmental Justice (EO 12898); Management and General Public Use of the National Wildlife Refuge System (EO 12996); Refuge Recreation Act of 1962, as amended; Invasive Species (EO 13112); Migratory Bird Treaty Act of 1918, as amended (MBTA); and Responsibilities of Federal Agencies to Protect Migratory Birds (EO 13186).

5.4 Distribution and Availability The draft Plan and EA will be made available for comment at a later date. A 30-day public notice will be placed in a regional newspaper. Readers will be directed to the Refuge website where they can read the EA, or request a hardcopy of the EA by phone or email. Comments will be accepted in writing either through email or regular post. Any comments or information received will be reviewed and changes incorporated into the Plan and EA, as appropriate.

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GLOSSARY

Action Threshold. Mosquito population levels that trigger integrated pest management (IPM) actions to manipulate mosquito populations.

Adulticide. Killing adult mosquitoes or a pesticide that kills adult mosquitoes.

Approved Acquisition Boundary. Area within which the Service is authorized to work with willing landowners to acquire and/or manage land. An approved acquisition boundary only designates those lands which the Service has authority to acquire and/or manage through various agreements, based upon planning and environmental compliance processes. Approval of an acquisition boundary does not grant the Service jurisdiction or control over lands within the boundary, and it does not make lands within the acquisition boundary part of the National Wildlife Refuge System. Lands do not become part of the National Wildlife Refuge System unless they are purchased or are placed under an agreement that provides for management as part of the Refuge System.

Arbovirus. Arthropod-borne virus. A viral disease carried and transmitted by mosquitoes or other arthropods.

Biological Diversity. The variety of life and its processes, including the variety of living organisms, the genetic differences among them, and communities and ecosystems in which they occur. (See 601 FW 3 for more information on biological diversity.)

Biological Integrity. Biotic composition, structure, and functioning at genetic, organism, and community levels comparable with historic conditions, including the natural biological processes that shape genomes, organisms, and communities. (See 601 FW 3 for more information on biological integrity.)

BMPs. Best management practices

CD. Compatibility Determination

Competence (mosquito). The relative ability of a mosquito species to carry and transmit virus.

CWA. Clean Water Act (33 U.S.C. 1251-1387)

CX. Categorical exclusion.

EA. Environmental assessment.

EAS. Environmental action statement.

EIS. Environmental impact statement.

Environmental Health. Composition, structure, and functioning of soil, water, air, and other abiotic features comparable with historic conditions, including the natural abiotic processes that shape the environment. (See 601 FW 3.)

Enzootic. A relatively consistent prevalence of disease in animals. The term is comparable to endemic, but refers to animals.

EPA. Environmental Protection Agency.

Epizootic. An outbreak of disease affecting many animals of one kind at the same time; also, the disease itself

ES. Ecological Services Program.

ESA. Endangered Species Act (16 U.S.C. 1531-1544).

Health Threat. An adverse impact to the health of human, wildlife, or domestic animal populations from mosquito-borne disease identified and documented by Federal, State, and/or local public health authorities. Health threats are locally derived and are based on the presence of endemic or enzootic mosquito-borne diseases, including the historical incidence of disease, and the presence and abundance of vector mosquitoes. Health threat levels are based on current monitoring of vectors and mosquito- borne pathogens.

Integrated Pest Management (IPM). A sustainable approach to managing pests by combining biological, cultural, physical, and chemical tools in a way that minimizes economic, health, and environmental risks.

Larvicide. Killing mosquito larvae, or a pesticide that kills mosquito larvae. MAD: Mosquito abatement district.

MMP. Mosquito Management Program

Monitoring (mosquito). Activities associated with collecting quantitative data to determine mosquito species composition and to estimate relative changes in mosquito population sizes over time.

Mosquito management. Any activity designed to inhibit or reduce populations of mosquitoes in the family Culicidae. Activity includes physical, biological, cultural, and chemical means of population control directed against any life stage of mosquitoes.

Mosquito-borne disease. An illness produced by a pathogen that mosquitoes transmit to humans and other vertebrates. The major mosquito-borne pathogens presently known to occur in the United States that are capable of producing human illness are the viruses causing eastern equine encephalitis, western equine encephalitis, St. Louis encephalitis, West Nile encephalitis/fever, LaCrosse encephalitis, and dengue, as well as the protozoans causing malaria.

NEPA. National Environmental Policy Act (42 U.S.C. 4321-4347).

NOAA. National Oceanic and Atmospheric Administration.

NOI. Notice of Intent.

Nontarget Organisms. Species or communities other than those designated for population control. NPDES. National Pollutant Discharge Elimination System established by section

402 of the NWRS. National Wildlife Refuge System.

Public Health Authority. A Federal, State, and/or local agency that has health experts with training and expertise in mosquitoes and mosquito-borne diseases and that has the official capacity to identify health threats and determine health emergencies.

PUP. Pesticide Use Proposal.

Pupacide. A pesticide that kills the pupal stage of mosquitoes.

PUR. Pesticide Use Report.

Refuge-Based Mosquitoes. Mosquitoes that are produced within, or occur on, a Refuge.Reservoir Host. A species in which a pathogen is maintained over time. Reservoir hosts are capable of transferring the pathogen to a vector.

Service. United States Fish and Wildlife Service

Service-Authorized Agent. A contractor, cooperating agency, cooperating association, Refuge support group, volunteer, or other party working on a Refuge on behalf of the Service to help achieve the Refuge purpose(s) or NWRS mission.

SLEV. Saint Louis encephalitis virus

Surveillance (mosquito-borne disease). Activities associated with detecting pathogens causing mosquito-borne diseases, such as testing adult mosquitoes for pathogens or testing reservoir hosts for pathogens or antibodies.

ULV. Ultra-low volume.

Vector. An organism, such as an insect or tick, that is capable of acquiring and transmitting a disease-causing agent, or pathogen, from one vertebrate host to another, or the act of transmitting a pathogen in such a manner.

Viremia. Level of virus in the blood.

WEEV. Western equine encephalomyelitis virus

WNV. West Nile Virus.

APPENDIX A. USFWS Director’s Memorandum dated May 27, 2014, Mosquito Management on National Wildlife Refuges

APPENDIX B. CT MMP – Mosquito Management an Integrated Approach

Connecticut Mosquito Management Program Mosquito Management An Integrated Approach

The Wetland Habitat and Mosquito Management (WHAMM) Program of the DEEP's Wildlife Division uses an integrated approach to manage mosquitoes that includes larval (immature) and adult mosquito population monitoring, public education, and cultural, biological and chemical control methods. Similar components are used in other Integrated Pest Management (IPM) strategies. However, the mosquito control profession has been practicing IPM concepts for over 100 years and has refined these practices to be specific to controlling mosquitoes. The American Mosquito Contro l Association and others recognize this specialized strategy as Integrated Mosquito Management or IMM.

Public education which promotes eliminating sources of mosquitoes around the home and minimizing exposure to mosquito bites by taking protective measures is critical to managing mosquitoes. Control measures can be initiated when immature (larval) mosquito levels reach certain threshold limits. Additional steps can be taken to reduce adult mosquito populations when viruses like Eastern Equine Encephalitis (EEE) or West Nile Virus (WNV) are detected in mosquitoes. If warranted, biological or chemical insecticides can be strategically applied by ground or aerial application equipment to control larvae or adults. Long-term control using water management, particularly in tidal wetlands, can be used for managing mosquitoes and can be further integrated into enhancing and restoring degraded wetland habitats. These various components of Connecticut's IMM strategy are discussed in further detail below.

Mosquito Management Around the Home The Connecticut Public Health Code prohibits homeowners from creating or maintaining sources of mosquitoes on their property. Violators are subject to enforcement actions by their local health department. There are several ways homeowners can minimize the number of biting mosquitoes in their yards. One of the easiest and surest ways to manage mosquitoes around the home is to eliminate standing water where mosquitoes can lay eggs. Mosquitoes need at least 7-10 days in water to fully develop. Some common sources of mosquitoes around the home are:

• Artificial containers that hold water (e.g., pails, paint cans, discarded tires pdf) • Open cesspools or septic tanks • Boat or pool covers or tarps that collect rain water • Unmaintained bird baths or wading pools • Storm sewer catch basins, rain barrels and clogged roof gutters • Rot holes in trees and stumps

Practice good sanitation around the home. Homeowners should properly dispose of or recycle trash which can hold rainwater. Make it a practice to flush bird baths and wading pools weekly. Swimming pool filtering systems should be maintained and in good working order. Abandoned pools should be drained, filled or "shocked" with pool chemicals. Openings for standing water sources, such as septic tanks or rain barrels, can be sealed or covered with screening. Rotten stumps and tree holes can be filled with sand. Discarded tires should be disposed of properly, holes (0.5 inches or larger) can be drilled in the bottom of the tires to drain rainwater or the tires can be stacked and covered to prevent rainwater from entering. Lawns and gardens should be watered minimally to prevent puddling and to conserve water.

Ornamental pools and aquatic gardens can become sources of mosquitoes if the water is allowed to stagnate. Water should be changed frequently or an aerator can be installed. Homeowners can practice their own biological control by stocking minnows, such as Gambusia, koi or guppies, which will eat mosquito larvae. The fish will need to be brought indoors for the winter or restocked annually because they will not survive Connecticut winters. Large pond stocking with non-native fish or releasing fish into public waters is prohibited. Insecticides, such as those containing Fusco the bacteria Bacillus thurgiensis var. israelensis (Bti), are available at many nurseries and garden supply centers and can be used to treat mosquito breeding sites. In general, natural ponds and lakes are not sources of mosquito breeding, because permanent bodies of water usually contain fish and other predators that would consume mosquito larvae.

There are also ways homeowners can minimize the annoyance caused by adult mosquitoes. Mosquitoes prefer to rest in shady, calm areas and will avoid more open sunny, breezy areas. Mowing tall grass will reduce places where mosquitoes can rest. Mosquitoes are most active around dawn and dusk although some, such as the common saltmarsh mosquito, may be active throughout the day or may be more active during cloudy, humid weather. Simply avoiding outdoor activity during these peak mosquito times can minimize contact with mosquitoes.

To reduce the chance of being bitten when outside, wear protective clothing such as long sleeves, long pants and head cover. Light-colored, loose-fitting clothing is preferable because dark clothing radiates more heat and attracts more mosquitoes. Insect repellents containing DEET, picaridin, IR3535 or oil of lemon eucalyptus can be used by most people and are often effective for varying lengths of time. Permethrin, a synthetic parathyroid that is widely available for repelling and killing ticks, also repels and kills mosquitoes. It is applied to clothing and provides longer-lasting protection. Do not apply permethrin products directly to skin. Although not marketed as repellents, there are several cosmetic liquids and creams that claim some level of mosquito repellency. These products may effectively repel when mosquito pressure is light, but need to be reapplied frequently. The U.S. Environmental Protection Agency (EPA) provides further information on the use and effective use of repellants.

Homeowners may also consider spraying pesticides labeled for mosquito control to shade trees, hedges and shrubs adjacent to foundations, fences and stone walls where adult mosquitoes are most likely to light. There are several over-the-counter aerosol sprays that homeowners can use to control mosquitoes. Always read and follow the label. Private, CT-certified applicators can also be hired to treat yards and neighborhoods. Make sure the applicator is certified in the "Mosquito and Biting Fly" category (cat. 7f) by the CT DEEP Pesticide Management Program. (Businesses Registered to Perform Mosquito Control in Connecticut)

To reduce mosquito infestations in the house, maintain screens over doors and windows. A porch or deck also can be enclosed with screening. Outside light use should be reduced and yellow light bulbs used when possible.

EEE and WNV can be fatal to horses. Horse owners are strongly encouraged to protect their horses from EEE and WNV by inoculation. Canine heartworm (filariasis) is a fatal disease circulated in dogs by biting mosquitoes. The DEEP, CAES or DPH do not monitor for heartworm in mosquitoes. Dog owners are encouraged to protect their pets from canine heartworm by administering preventative medications obtained through their veterinarian.

Managing Mosquitoes Using Insecticides Insecticides used for mosquito management are grouped into two categories. Larvicides/pupacides are used to control immature (larval or pupal) mosquitoes in aquatic habitats. Adulticides are used to control adult mosquitoes. The insecticides used are registered by the EPA and the CT DEEP Pesticide Management Program and do not pose any health hazards to humans or the environment when used in accordance with the label.

Larvicides are applied by hand-, backpack or aerial application equipment to mosquito-breeding habitats when there is an abundance of larvae. Larviciding is more efficient and effective for managing mosquitoes than adulticiding because the larvae are concentrated in relatively small, well-defined, aquatic habitats. If larval control methods are successful, the need for adult mosquito management is greatly reduced or eliminated.

Currently, the primary larvicides used by the WHAMM Program are the microbial compounds Bti (Bacillus thuringiensis var. israelensis) and Bs (B. sphaericus) and insect growth regulators containing methoprene. The microbial products release toxins when ingested by the filter-feeding mosquito larvae. Bti and Bs target mosquitoes and Bti is also labeled to control black flies and some midges. Bti has a short effective life (two to three days) and must be reapplied to each new generation of mosquitoes. The bacterial spores in Bs recycle in the larval mosquito population and can provide 4-6 weeks of larval control. Since these products must be ingested by mosquito larvae to be effective, they do not control the non-feeding pupae or adult mosquitoes.

Methoprene is a compound that mimics the action of an insect growth-regulating hormone and prevents the normal maturation of mosquito larvae. The active ingredient, S-methoprene, breaks down rapidly in

ultraviolet light. Methoprene does not persist in the environment but encapsulated formulations allow a slow release of minute amounts of methoprene and can provide several weeks of control. Methoprene is also used in flea and tick control in pets.

Larvicide Plan (Strategies for the Application of Larvicides to Control Mosquitoes in Response to West Nile Virus in Connecticut - Supplement to West Nile Virus Response Plan)

Pupacides in the form of monomolecular films (MMF's) or oils create a thin film on the water surface which drowns the larvae, pupae or emerging adult. MMF's break down in about 10-14 days.

Adulticides are considered for use by the WHAMM Program for reducing the adult mosquito population when a public health threat from mosquito-borne diseases like EEE or WNV exists. Adulticiding provides an immediate but short-term reduction in adult mosquito numbers. Backpack or truck-mounted equipment is used to create tiny, ultra-low volume (ULV) droplets of insecticide that drift through the swarm of mosquitoes or impinge on vegetation on which the mosquitoes will land. Truck-mounted applications are used in relatively small, localized areas where road access allows adequate coverage. If, however, the public health threat exists in a larger geographic area, where truck-mounted spraying would be ineffective, aircraft can be used to aerially-apply adulticides. The primary adulticides used by the WHAMM Program contain synthetic pyrethroids, such as resmethrin, sumithrin, or bifenthrin. These products contain the same active ingredients as several over-the-counter yard, garden and pet sprays. They do not pose unreasonable risks to humans or the environment when applied according to the label. Adulticiding is more costly than larviciding because adulticides are usually applied over larger areas.

The WHAMM Program is actively evaluating new mosquito control products as they become available. New products must provide consistent mosquito control, be nonhazardous to humans and the environment and be cost-effective. If new products meet these requirements, they are considered for possible use.

Mosquito Control Using Water Management Where environmentally feasible, the WHAMM Program uses water management for source reduction and biological control of mosquitoes by making the sites 1) unsuitable for mosquito egg and larval development and 2) enhancing the area to provide open water habitat for natural mosquito predators such as fish and birds. This method provides more permanent control of mosquitoes than insecticides, resulting in a substantial reduction in insecticide applications and costs.

In tidal saltmarshes, a technique known as Open Marsh Water Management (OMWM) is the preferred method for controlling mosquitoes and enhancing or restoring wetland habitat (more about OMWM pdf). Unlike the parallel grid-ditch method used in Low ground pressure equipment is used to the 1930's which had adverse affects on tidal wetland hydrology excavate shallow pools and channels on the marsh and habitat, OMWM involves the selective excavation of shallow smface to control mosquitoes and provide wildlife pools and ditches in mosquito-breeding areas. These pool and habitat. ditch networks are not connected directly to tidal channels and, therefore, do not drain at low tide. A higher water level is maintained in the pools which provides habitat for fish and other wildlife and encourages revegetation of the surrounding marsh by native grasses. Mosquito management is achieved by modifying egg-laying sites and by creating open water habitat for small naturally-abundant killifish, which prey on mosquito larvae and pupae. OMWM systems provide long-term control of mosquitoes, thus reducing the need to apply insecticides.

Mosquito Myths A number of products on the market claim to have mosquito control capabilities. In most cases, these products have not been rigorously tested and do not perform as advertised. Mechanical traps, such as ultraviolet "bug zappers" or devices that repel using Shallow pools and plugged ditches restore ultrasonic sound waves, do not meet advertiser's claims. In fact, wildlife habitat and control mosquitoes bug zappers attract few mosquitoes and may actually kill beneficial insect predators. The Connecticut MMP as well as other states and the American Mosquito Control Association do not endorse the use of these products to reduce mosquito infestations. Natural products, such as citronella-scented candles and plants, clove oil, peppermint, or diet supplements like garlic or vitamins that claim to repel mosquitoes are not supported by scientific evidence. There are many individuals who feel these products are effective; however, each person has a unique metabolism and body chemistry and these products may not be equally effective for everyone.

Natural predators, such as bats and some birds, will eat adult mosquitoes as do other types of insects such as dragonflies. However, studies have shown that mosquitoes make up a very small percentage of a bat or bird's diet. Bats and insectivorous birds are opportunistic feeders and may consume a large quantity of mosquitoes if mosquito populations are very high. However, if adult mosquitoes are at moderate or low levels (but yet are at pestiferous levels or in numbers that could still effectively transmit disease) bats or birds will not expend the energy to chase enough mosquitoes to obtain the equivalent amount of food as say a moth or large beetle. The CT Mosquito Management Program encourages the placement of bat and bird houses for the conservation of these species but does not endorse the use of Open marsh water management creates and them solely for the control of mosquitoes. For further information restores habitat for fish, inveitebrates. on mosquito myths read the Rutgers Cooperative Research and shorebirds, wading birds and waterfowl Extension Fact Sheet (pdf) or visit the University of Florida's Mosquito Information Website .

Content last updated February 20

APPENDIX C. USFWS Compatibility Determination

JUSTIFICATION FOR A FINDING OF APPROPRIATENESS OF A REFUGE USE

Refuge Name: Great Meadows Unit at Stewart B. McKinney National Wildlife Refuge

Use: Mosquito Management

NARRATIVE: Mosquito management includes population monitoring and control, if warranted. Mosquito surveillance monitoring and control, when necessary, will be conducted in 5 acres of wetlands within the Great Meadows Unit of the refuge. Mosquito and mosquito-borne disease management is not a priority public use of the National Wildlife Refuge System (NWRS) under the National Wildlife Refuge System Administration Act of 1966 (16 U.S.C. 668dd-668ee), as amended by the National Wildlife Refuge System Improvement Act.

The Connecticut Mosquito Management Program (CT MMP) will conduct mosquito population monitoring and control, following the protocols and best management practices identified in the Connecticut West Nile Virus Surveillance and Response Plan (CT DEEP 2012) and in compliance with refuge-specific regulations. In general, we allow populations of native mosquito species to function unimpeded unless they cause a wildlife or human health threat. Mosquitoes are a natural component of most wetland ecosystems but may also represent a threat to human, wildlife, or domestic animal health. Refuge personnel are to collaborate with Federal, State, or local public health authorities and vector control agencies to identify refuge-specific health threat categories that represent increasing levels of health risks that are based on monitoring data.

Mosquito-associated health threats will be addressed using an integrated pest management (IPM) approach, including when practical, compatible, non-pesticide actions that reduce mosquito production. Treatment options will be chosen based on our IPM policy (569 FW 1) and the Biological Integrity Diversity and Environmental Health (BIDEH) policy (601 FW 3), and will emphasize human safety and environmental integrity, effectiveness, and cost factors. We will use human, wildlife, or domestic animal mosquito-associated health threat determinations, combined with refuge mosquito population estimates, to determine the appropriate refuge mosquito management response. We will allow pesticide treatment to control mosquitoes on refuge lands only after evaluating all other reasonable IPM actions. The decision to use pesticide treatments will be based on monitoring data for the relevant mosquito life stage and used only when necessary to protect the health of human, wildlife, or domestic animals. Mosquito surveillance activities would be conducted from April through October under the conditions of this compatibility determination and special use permit (SUP) in accordance with the Director’s May 27, 2014 Order and the BIDEH policy.

COMPATIBILITY DETERMINATION

USE: Mosquito Management

REFUGE NAME: Great Meadows Unit at the Stewart B. McKinney National Wildlife Refuge

DATE ESTABLISHED: February 25, 1985

ESTABLISHING AND ACQUISITION AUTHORITY(IES): Stewart B. McKinney National Wildlife Refuge (NWR; refuge), formerly the Connecticut Coastal NWR, was authorized for re-designation by an Act of Congress under Title II of the Wetlands Loan Extension Act (Public Law 98-548) on October 26, 1984. The refuge is managed by the U.S. Fish and Wildlife Service (Service; USFWS).

The legislation designated four lands, totaling 151 acres, for initial acquisition including Milford Point, Chimon, Sheffield, and Falkner Islands. The refuge was initially established on February 25, 1985, with the acceptance of the deed to Chimon Island by the U.S. Fish and Wildlife Service (Service) from The Nature Conservancy of Connecticut. On May 13, 1987 (P.L. 100-38) the refuge was re-designated as the Stewart B. McKinney NWR in honor of Congressman McKinney’s efforts in its establishment.

The Connecticut Coastal Protection Act authorized the expansion of the refuge on October 19, 1990, by incorporating Salt Meadow NWR as a unit of the Stewart B. McKinney NWR and permitting future land acquisitions (P.L. 101-443, H.R. 3468). The Salt Meadow NWR was established in 1971 under authority of the Migratory Bird Conservation Act of 1934, as amended. The refuge currently consists of ten management units: Salt Meadow, Falkner Island, Sheffield Island, Milford Point, Chimon Island, Great Meadows, Goose Island, Outer Island, Calf Island, and Peach Island.

REFUGE PURPOSE(S):

• “… to conserve (A) fish or wildlife which are listed as endangered species or threatened species …. Or (B) plants …”16 U.S.C. § 1534 (Endangered Species Act of 1973)

• “… suitable for− (1) incidental fish and wildlife-oriented recreational development, (2) the protection of natural resources, (3) the conservation of endangered species or threatened species …” 16 U.S.C. § 460k-1“… the Secretary … may accept and use … real … property. Such acceptance may be accomplished under the terms and conditions of restrictive covenants imposed by donors …” 16 U.S.C. § 460k-2 (Refuge Recreation Act (16 U.S.C. § 460k-460k-4, as amended).

• “… particular value in carrying out the national migratory bird management program.”16 U.S.C. § 667b (An Act Authorizing the Transfer of Certain Real Property for Wildlife, or other purposes)

• “… for use as an inviolate sanctuary, or for any other management purpose, for migratory birds.” 16 U.S.C. § 715d (Migratory Bird Conservation Act)

• “... to protect the natural features of a contiguous wetland area.” 84 Stat. 1095, dated Oct. 22, 1970.

NATIONAL WILDLIFE REFUGE SYSTEM MISSION: The mission of the National Wildlife Refuge System is to administer a national network of lands and waters for the conservation, management, and where appropriate, restoration of the fish, wildlife, and plant resources and their habitats within the United States for the benefit of present and future generations of Americans. (National Wildlife Refuge System Improvement Act of 1997, Public Law 105-57; 111 Stat. 1252).

DESCRIPTION OF USE:

(a) What is the use? Is the use a priority public use? The use is to implement an Integrated Pest Management (IPM) plan that consists of a phased approach to mosquito management, which includes population monitoring and, if warranted, control on the Great Meadows Unit at the refuge. The Service uses regulations and policies to plan and guide mosquito management actions on refuges. Enabling regulations of the Refuge System are contained in Title 50 Code of Federal Regulations (CFR) Subchapter C, Part 25, Administrative Provisions, as well as the National Wildlife Refuge System Administration Act of 1966, as amended by the National Wildlife Refuge System Improvement Act of 1997, as amended (16 U.S.C. §§ 668dd-668ee). Guiding policies for mosquito management are referenced in a USFWS Director’s Memorandum dated May 27, 2014 and include the:

• Comprehensive Conservation Planning Process (602 FW 3)

• Step-Down Management Planning (602 FW 4)

• Biological Integrity, Diversity, and Environmental Health (601 FW3)

• Integrated Pest Management (569 FW 1)

• Appropriate Refuge Uses (603 FW 1)

• Compatibility (603 FW 2)

Mosquito and mosquito-borne disease management is not a priority public use of the National Wildlife Refuge System under the National Wildlife Refuge System Administration Act of 1966 (16 U.S.C. 668dd-668ee), as amended by the National Wildlife Refuge System Improvement Act of 1997.

The Connecticut Mosquito Monitoring Program (CT MMP) will conduct mosquito population monitoring and control, following the protocols and best management practices identified in the Connecticut West Nile Virus Surveillance and Response Plan (Connecticut Department of Energy and Environmental Protection [CT DEEP] 2012) and in compliance with refuge-specific regulations. The Service recognizes that mosquitoes are a natural component of most wetland ecosystems but may also represent a threat to humans, wildlife, or domestic animal health. Refuges are to collaborate with Federal, State, or local public health authorities and vector control agencies to identify refuge-specific health threat categories that represent increasing levels of health risks and are based on monitoring data. Mosquito-borne arboviruses of public health importance in Connecticut include West Nile Virus (WNV), Eastern equine encephalitis (EEE), Jamestown Canyon (JC), Cache Valley (CV), Trivittatus (TVT), Highlands J (HJ), LaCrosse (LAC), and Potosi (POT) viruses (CT DEEP 2012).

The refuge will not conduct mosquito monitoring or control, but may allow these activities under a special use permit (SUP).

(b) Where would the use be conducted? Mosquito surveillance monitoring and control, if necessary, will be conducted in 5 acres of salt marsh and freshwater wetlands within the Great Meadow Unit of the refuge.

Currently, the Connecticut Agricultural Experiment Station (CAES) maintains a network of 91 fixed mosquito-trapping stations located in 72 municipalities throughout the state. Two stations are located within one mile of the refuge’s Great Meadow Unit: the Beacon Point site (#ST64) in Stratford and the Barrum Boulevard site (#BP40) in Bridgeport. Connecticut has 52 species of mosquitoes, many of which can be found on the refuge or in the Stratford/Bridgeport area; these include both freshwater and saltwater varieties (Wolfe 2015 personal communication). Some species, such as the salt marsh species, are aggressive daytime biters and can fly up to 15 miles in search of a blood meal. Many are also involved in the amplification and transmission of diseases, such as EEE and WNV. Each year, mosquito traps in Stratford, Bridgeport and surrounding towns that are sampled by the CAES frequently contain mosquitos infected with WNV. The highest levels of WNV (in mosquitoes and humans) in 2014 were found near the refuge’s Great Meadow Unit (Figure 1). The WNV has been identified in 21 mosquito species identified in Connecticut, with 72 percent of the virus isolates from Culex pipiens (http://www.ct.gov/dph/lib/dph/infectious_diseases/ctepinews/vol34no2.pdf).

From at least 1998 through 2007, the CT MMP conducted mosquito surveillance and applied larvicide to less than 15 acres of the Great Meadow Unit until the practice was suspended pending review of the Service’s new compatibility process (Wolfe 2015 personal communication; Figure 2). The CT DEEP continues to annually identify the Great Meadows Unit as an area of concern to public health because of the risk from mosquitoes (Figure 3).

Figure 1. 2014 West Nile Virus Activity in Connecticut. (http://www.ct.gov/caes/cwp/view.asp?a=2819&q=377446&caesNav=%7C; accessed January 2015)

Stewart B. McKinney NWR Great Meadow Unit

Town Trap Site Number of Positive or WNV, EEE, JC Mosquito Species Mosquitoes Negative (Number of Mosquitos) Bridgeport Barnum Blvd. 3,168 Positive WNV: 4 Cx. pipiens: 3 Cx. salinarius: 1 Stratford Beacon 5,009 Positive WNV: 20 Cx. pipiens: 18 Point Ae. vexans: 1 Oc. trivittatus: 1 Year to Date Totals for 229,097 WNV(68) All 91 Trap Sites JC (23)

Figure 2. Areas of public health concern in the Great Meadows Unit of Stewart B. McKinney NWR for 2003 (CT DEEP 2003).

Figure 3. Areas of public health concern in the Great Meadows Unit of Stewart B. McKinney NWR for 2014 (CT DEEP 2014).

(c) When would the use be conducted? Surveillance activities associated with this use would be conducted on the Great Meadows Unit once per week from April through October by CT MMP staff under the conditions of this compatibility determination and a special use permit. Known sites for mosquito production on the refuge will be identified for monitoring and surveillance during periods of mosquito production. The timing and frequency of monitoring will be based on a number of factors, including history of mosquito production, tidal cycles, precipitation levels, and available resources. In Connecticut, mosquito larvae/pupae need at least 5 to 7 days in water to fully develop; they cannot survive in situations where surface water drains off or evaporates within that time frame. Therefore, inspections will focus on identifying and sampling shallow depressions, occluded ditches, or similar sites in tidal and non-tidal habitat that could hold tidal or rain water for more than 5 to 7 days. Inspections will occur during normal working hours.

Mosquito control activities will only occur outside of normal working hours (irregularly) when: 1) it is necessary to protect the health and safety of humans, wildlife, or domestic animals, and 2) after surveillance monitoring data show that mosquito populations have reached specified abundance criteria.

The treatment of marshes on the Great Meadows Unit of the refuge using larvicides would only be permitted after the CT MMP has provided the refuge manager with information as described in the Connecticut West Nile Virus Surveillance and response Plan (CT DEEP 2012) that shows mosquito larvae populations are widespread within the salt marsh (areas C and D), and after monitoring data reveals that mosquito populations meet or exceed the following abundance measures: using a long- handle, plastic, 8 ounce cup dipper - if 5 or more larvae/dip are found in 50 percent or more of the dips (minimum of 3 dips/site). Other factors to be considered in making the determination to allow larvacide treatment include marsh hydrology (drying versus flooding), rainfall, temperature, instar larval stages, and treatment history.

Pupacides or adulticides will only be used when large numbers of mosquitoes are considered an immediate threat to human health and thresholds developed by the appropriate CT MMP met or exceeded (i.e., there is active transmission of mosquito-borne disease on the Great Meadows Unit of the refuge from refuge-based mosquitoes).

(d) How would the use be conducted? Mosquito-associated health threats will be addressed using an integrated pest management (IPM) approach including, when practical, compatible, non-pesticide actions that reduce mosquito production as found in the Mosquito Management Plan for the Great Meadows Unit at the refuge (USFWS 2017). We will choose treatment options based on our IPM policy (569 FW 1) and our Biological Integrity Diversity and Environmental Health policy (601 FW 3). We will base the choice on and emphasize: human safety and environmental integrity, effectiveness, and cost. We will use human, wildlife, or domestic animal mosquito-associated health threat determinations combined with refuge mosquito population estimates to determine the appropriate refuge mosquito management response. We will consider allowing pesticide treatment to control mosquitoes on refuge lands after we have evaluated all other reasonable IPM actions.

In Connecticut, mosquito control activities and work are performed pursuant to the provisions of the State’s General Statutes (http://www.ct.gov/mosquito). The CT MMP monitors larval and adult mosquitos throughout the state and adheres to the Connecticut West Nile Virus Surveillance and Response Plan (CT DEEP 2012). Additionally, the CT MMP will conduct surveillance, monitoring,

and if necessary, control measures, and the monitoring of the efficacy to control the targeted life stage under the conditions contained in a SUP that will be issued by the refuge manager.

Baseline mosquito management actions on the Great Meadows Unit at Stewart B. McKinney NWR will involve monitoring and surveillance of mosquito vector populations. Annual surveillance monitoring on refuge lands for arbovirus incidence in adult mosquito vectors and wildlife (especially birds) will be allowed. Mosquito vector monitoring on the refuge will document mosquito species composition to genus or species level, and estimate population size and distribution across refuge wetland habitats during the breeding season, using standard methods employed by mosquito control professionals.

Mosquito population monitoring objectives are to:

• Establish baseline data on species and abundance.

• Map breeding and harboring habitats.

• Estimate relative changes in population sizes and evaluate associated health risks for making IPM decisions to reduce mosquito populations when necessary

• Determine the effectiveness of treatment.

• Use this information to guide integrated pest management of mosquito populations.

All sites identified as potential mosquito habitat have been logged and recorded in the CT MMP GIS system. Throughout the mosquito season, CT MMP staff conducts larval surveys every 10 days at each site on a regular rotation from June through October. The CT MMP checks all sites known to harbor mosquitoes for mosquito larvae using a standard (8 oz.) dipper and may search for new larval habitats on or adjacent to the Great Meadows Unit of the refuge. Two trap types are used at all trapping stations. A carbon dioxide light trap is designed to trap host-seeking adult female mosquitos of any species. A gravid mosquito trap with hay infusion is designed to trap previously blood-fed adult female mosquitos, principally Culex species and container-breeding Ochlerotatus species. When the traps are deployed, adult mosquitoes are collected from them weekly, taken back to the laboratory each morning, identified to the species level, and counted. A maximum of 50 female mosquitos are included in the pooled data, and aliquots of each mosquito pool are inoculated into Vero cell cultures for detection of WNV and other mosquito-borne arboviruses of public health importance. Isolated viruses are identified by Real Time (TaqMan) PCR or standard RT-PCR using virus-specific primers, plaque reduction neutralization (PRNT) and/or an enzyme-linked immunosorbent assay (ELISA) with specific reference antibodies. All of the virus isolation work is conducted in a certified Bio-Safety Level 3 laboratory at the CAES. Weekly test results are reported to the Centers for Disease Control and Prevention electronically via ArboNet and to the DPH for dissemination to other state agencies, local health departments, the media, and neighboring states.

Monitoring and control will be conducted by the CT MMP, primarily on foot. Vehicle access to the marsh edge will be restricted to established roads and fire trails at the refuge. To avoid harm to wildlife or habitats, access to traps and sampling stations will comply with the Stipulations Necessary to Ensure Compatibility included in this determination.

Refuge staff has coordinated with the CT MMP to develop a mosquito management plan that will provide specifics on how and when the refuge will allow, if necessary, control of mosquitoes on

refuge lands, using predetermined threat levels and mosquito vector population densities (Table 1). A phased approach will be used to guide appropriate control response up to and including the use of adulticides (table 2). That will occur when Federal and State public health officials, using arbovirus monitoring and surveillance data, have determined that the Great Meadows Unit at the refuge is in a high- risk area for mosquito-borne disease transmission, and it has been demonstrated through surveillance that refuge-based mosquitoes have been shown to carry specific diseases. A high-risk determination indicates an imminent risk of serious human disease or death.

Pesticide treatment may not be used on the Great Meadows Unit at Stewart B. McKinney NWR solely for nuisance mosquito relief, but may be considered when there is a demonstrated human health risk, mosquitoes are detrimental to refuge goals and objectives, and mosquito management actions will not interfere with refuge goals and objectives. Only pesticides identified in the special use permit and for which a pesticide use proposal (PUP) has been submitted and approved will be used on the refuge. The preferred larvicide treatments for use on the Great Meadows Unit at the refuge are Bacillus thuringiensis israelensis (Bti) or Bacillus sphaericus (Bs), because of the bacterium’s limited non-target effects. Due to specificity of the effects of Bti and Bs on the insect order Diptera, VectoBac® GS is deemed compatible for use, under the stipulations prescribed at the end of this compatibility determination. Bti and Bs are the preferred chemical control option and will be used under appropriate conditions. We favor using the larvicide that would have the least adverse impacts on non-target invertebrates, produce fewer disruptions to food webs critical for migratory birds, and reduce lethal effects on natural mosquito predators, such as larval forms of odonates, hemipterans, and coleopterans. CT MMP will conduct post-larvicide monitoring to determine effectiveness.

Table 1. The refuge’s phased mosquito management response to mosquito threat level.

Threat Condition Response Level

1 No documented existing health threat. Monitoring and surveillance of areas Mosquito management issues have not surrounding the refuge to inform been reported or identified by the CT management actions on the refuge. MMP. Remove/manage artificial breeding sites such as tires, tanks, or similar debris/containers. Consult with CT MMP when planning wetland enhancement or restoration projects. 2 Potential human or wildlife (including Response as in threat level 1 plus: allow threatened and endangered species) compatible monitoring and disease health threat (presence of vector surveillance. Consider compatible non- species, historical health threat, etc.), as pesticide management options to reduce documented by the CT MMP. the potential for above-normal mosquito production (e.g., restore/enhance tidal marsh hydrology). 3 Mosquito larvae threshold2 exceeded Response as in threat level 2 plus: allow for human and/or wild/domestic animal compatible site-specific application of health on the refuge as determined by larvicide in areas with above average standardized monitoring. Documented mosquito populations as determined by potential human or wild/domestic monitoring. Conduct post larvicide animal health threat (historic health monitoring to determine efficacy threat, presence of vector species).

Threat Condition Response Level

4 Mosquito larvae have begun to reach Response as in threat level 3 plus: if last instar stages or pupate reducing the appropriate, increase the intensity and efficacy of larvicides. Mosquto larval frequency of larvicides allow compatible and pupal population thresholds site-specific use of pupacides in areas with exceeded on the refuge. Mosquitoes above average mosquito populations, produced by the refuge pose a health determined through monitoring to be threat as determined by the CT MMP. beyond control with larvicides. Increase Before the use of pupacides, the monitoring and disease surveillance. Directors of the CT DEEP and CT DPH Conduct post larvicide and pupacide will approve the use in coordination monitoring to determine efficacy. with the Refuge Manager. This approval will be documented by the CT DEEP. 5 Exceedance of larval, pupal, and adult Response as in threat level 4 plus: mosquito population thresholds2 on the consider site specific adulticiding (with a refuge. High risk for mosquito-borne pyrethrin-based adulticide) in areas with disease (imminent risk of serious above average mosquito populations as human disease or death, or an imminent determined by monitoring. Conduct post risk of serious disease or death to adulticide monitoring to determine populations of wild/domestic animals) efficacy. within communities surrounding the refuge has been documented by the CT MMP and a declaration of a Public Health Emergence issue by the Governor of the State of Connecticut.

2Human health threshold (e.g., numbers per dip) is determined by considering several factors as determined by Appendix F. Larval thresholds are presented in state plans in Appendix F

A description of the refuge’s phased approach to mosquito management is contained in the following:

In Phase 1 a health threat has not been identified and mosquito management issues have not been reported or identified by the CT MMP. To help prevent possible mosquito management issues, artificial mosquito breeding habitat throughout the Great Meadows Unit at the refuge, such as tires, open containers, and other equipment or objects that pool water where mosquitoes may breed, should be eliminated. Although artificial mosquito breeding habitat is not currently an issue on the Refuge, this should still be a concern into the future as a preventative measure.

The refuge would consult with the CT MMP when wetland enhancement or restoration projects are being planned on the refuge. Consultation would allow refuge staff and the CT MMP to identify potential issues and/or opportunities related to mosquito production and management in the future. The monitoring and surveillance of mosquito abundance and disease prevalence in areas similar to and near the Great Meadows Unit at the refuge would be conducted by the CT MMP, which would then inform of the potential for mosquito management needs on the refuge.

In Phase 2 the refuge manager will be contacted by the CT MMP of the potential human health threat posed by mosquitoes harbored or produced on the refuge. In response, refuge staff may request increased mosquito population monitoring and disease surveillance by the CT MMP. The initial step to developing a proactive prevention and management program for mosquitoes is to determine mosquito species presence and abundance on the Great Meadows Unit at the refuge, and to identify potential or documented vectors of mosquito-borne diseases that represent a potential human health threat. This would be accomplished at the State monitoring locations – of which there are two – adjacent to the Great Meadows Unit. These stations will determine adult species type, abundance, and if the mosquiotes harbor disease vectors. Monitoring and surveillance activities should be well-documented and presented to Refuge staff by the CT MMP. This data will help determine the need to control larval mosquitoes.

Monitoring and surveillance activities should be well-documented and presented to refuge staff by the CT MMP. In order to avoid or minimize the use of pesticides, habitat management practices or wetland enhancement/restoration projects that improve wildlife habitat and reduce seasonal abundance of larval and adult mosquitoes should be implemented where possible. Refuge staff and visitors would be informed of an increased health threat associated with mosquito-borne disease activity. Personal protection measures such as wearing mosquito repellant would be recommended to staff and visitors.

In Phase 3 the application of larvicides would be considered if non-pesticide attempts to reduce mosquito populations are unsuccessful or are not feasible, and mosquito larvae thresholds have been exceeded. Locations of larvicide treatments would be based on standardized monitoring results. The preferred larvicide treatments are biorationals (biological agents) Bti, and Bs. because of limited non-target effects. Post larvicide monitoring would be conducted to determine efficacy

In Phase 4, the intensity and frequency of larvicides would be increased if appropriate. Larvicides (Bti and Bs, are only effective on mosquitoes during early instar stages (up to the fourth) and do not control pupae. If developing mosquitoes have reached the last instar stages or have pupated, then we would consider site-specific pupacides in areas with above average mosquito populations (determined through monitoring). Because pupacides can negatively affect all invertebrates that require surface air (e.g., act as surfactants), the use of these pesticides should be carefully considered. Due to the broad-spectrum action of surface oils and films, they typically are not appropriate and are rarely authorized for use on refuges. For this reason, pupacides (Agnique) would only be used if large numbers of mosquitoes are considered an immediate threat to human health and thresholds developed by the appropriate public agency have been exceeded (there is active transmission of mosquito-borne disease from refuge based mosquitoes or within flight range of vector mosquito species present on the Great Meadows Unit at the refuge). Post larvicide and pupacide monitoring would be conducted to determine efficacy and any adverse impacts. The use of pupacide on the Refuge would only be allowed after the Directors of CT DEEP and DPH have approve the use in coordination with the refuge and that determination as been documented by CT DEEP.

In Phase 5, mosquito-borne disease activity has been documented on the Great Meadows Unit at the refuge or within flight range of vector mosquito species present on the refuge. There is active transmission of mosquito borne disease on the Great Meadows Unit at the refuge from refuge based mosquitoes. A risk of serious mosquito-borne human disease or death has been documented by the appropriate public health authority. Disease surveillance determines that there is a high risk for

mosquito-borne disease within the vicinity of the refuge. For example, pathogen presence in mosquito pool(s), wild birds, sentinel chicken flock(s), horses, or humans has been documented within the flight range of vector mosquito species present on the refuge. These conditions in combination with adult mosquito populations above thresholds levels on the refuge would trigger consideration of a more aggressive treatment strategy, including the use of Pyrethrin-based adulticides. If larvicide and/or pupacide treatments fail, pyrethrin-based adulticides would be considered for use on the refuge to suppress populations of infected mosquitoes and interrupt epidemic virus transmission. Because the efficacy and effects of adulticides are variable, adulticides should not be applied broadly without site-specific data indicating a need for control. Further, the use of adulticide would be considered in concert with the Assessment found in the West Nile Virus Surveillance and Response Plan, Contingency Plan for Eastern Equine Encephalitis, and Zika Virus Surveillance and Response Plan and only after the Governor has issued a public health emergency. The CT MMP would be required to prepare a Risk Assessment as part of their request to apply adulticides. The Risk Assessment evaluates a number of factors including environmental conditions, species presence, virus infection rate, sentinel chicken seroconversion, dead bird presence, and human cases to determine whether adulticide should be considered. We would only consider application in areas where a pathogen is present and mosquito population thresholds have been exceeded on the refuge that can be effectively treated while minimizing non-target effects, especially to threatened and endangered species. However, specific areas treated and the extent of treatment would vary from year to year depending on mosquito populations and environmental conditions.

In order to limit human contact with adulticides, visitors would not be allowed in parts of the refuge that are being treated with adulticides. Information about treatment scheduling, location, and pesticide would be posted on the refuge website, at the refuge Headquarters, and at the treatment location. Post adulticide monitoring would be conducted to determine efficacy and any adverse impacts.

In summary, application of adulticides on the Great Meadows Unit at the refuge would require the following steps:

• Prior approval from the National IPM Coordinator via an approved Pesticide Use Proposal (PUP) • The CT MMP must present the refuge manager with data supporting presence of an arboviral disease on the refuge or within flight range of the vector mosquito species on the refuge, including a Risk Assessment in the region • The CT MMP must provide the refuge manager with the types/quantities of adulticides proposed as well as the proposed areas for treatment • If beneficial, the CT MMP should conduct simultaneous applications of larvicides with the adulticide application to prevent future adult outbreaks

Based on monitoring data, we will determine the most appropriate pesticide treatment options for the relevant mosquito life stage. We will use current monitoring data for larval, pupal, and adult mosquitoes to determine the need for larvicides, pupacides, and adulticides, respectively. Treatment areas will be based on surveillance and monitoring results. Specific areas treated and the extent of treatment would vary from year to year depending on mosquito populations, the mosquito vector flight distance, and environmental conditions. We will allow the use of adulticides only when the state declares a public health emergency and there are no practical and effective alternatives to reduce mosquito populations on the Great Meadows Unit of the refuge. We will not allow pesticide treatments for mosquito control on the Great Meadows Unit at the refuge without current mosquito population data indicating that such actions are warranted. We require an approved PUP prior to applying pesticides on the Great Meadows Unit at the refuge.

If appropriate, the intensity and frequency of larvicides would be increased. Larvicides (Bti and Bs,) are only effective on mosquitoes during early instar stages and do not control pupae. If developing mosquitoes have reached the last instar stages or have pupated, then we would consider site-specific pupacides in areas with above average mosquito populations (determined through monitoring). Because pupacides can negatively affect all invertebrates that require surface air (i.e., they act as surfactants), the use of these pesticides should be carefully considered. For this reason, pupacides would only be used if: 1) large numbers of mosquitoes are considered an immediate threat to human health; and 2) thresholds developed by the appropriate public agency have been exceeded; and 3) there is active transmission of mosquito-borne disease from refuge based mosquitoes, or within flight range of vector mosquito species present on the refuge. Post larvicide and pupacide treatment monitoring would be conducted to determine efficacy and any adverse impacts.

The following conditions, in combination with adult mosquito populations above specified thresholds levels on the refuge, would trigger consideration for a more aggressive mosquito treatment strategy, including the use of adulticides:

• If mosquito-borne disease activity has been documented on the Great Meadows Unit at the refuge or within flight range of vector mosquito species present on the refuge. • A risk of serious mosquito-borne human disease or death has been documented by the appropriate CT MMP. • Disease surveillance has determined that there is a high risk for mosquito-borne disease within the vicinity of the refuge. For example, pathogen presence in mosquito pool(s), wild birds, sentinel chicken flock(s), horses, or humans has been documented within the flight range of vector mosquito species present on the refuge.

If larvicide and/or pupacide treatments fail, pyrethrin-based adulticides would be considered for use on the refuge to suppress populations of infected mosquitoes and interrupt epidemic virus transmission. Because the efficacy and effects of adulticides are variable, adulticides would not be applied broadly without site-specific data indicating a need for control. Further, the use of adulticide would be considered in concert with the CT West Nile Virus Surveillance and Response Plan, Contingency Plan for Eastern Equine Encephalities, Zika Virus Surveillance and Response Plan, and only after the Governor of the State of Connecticut has declared a Public Health Emergency.

The CT MMP would be required to prepare a Risk Assessment as part of their request to apply adulticides. The Risk Assessment evaluates a number of factors including environmental conditions, species presence, virus infection rate, sentinel chicken seroconversion, dead bird presence, and human cases to determine whether adulticide should be considered. We would only consider

application in areas where a pathogen is present and mosquito population thresholds have been exceeded on the refuge that can be effectively treated while minimizing non-target effects, especially to threatened and endangered species.

Treatment regimens will vary annually, depending on the current threat level; the process for determining the threat level will be clearly delineated in the Mosquito Management Plan for the Great Meadows Unit at Stewart B. McKinney NWR. Because disease threat levels vary from year to year, mosquito management on the refuge is unlikely to include all phases in any given year. Action thresholds that trigger chemical interventions will incorporate various factors listed in Service Policy 601 FW 7, Exhibit 3, as developed with refuge staff, State mosquito control section, public human health services, and vector control agencies. Thresholds must be genus and life-stage specific and be related to the refuge decision-making response matrix.

We will rarely allow the CT MMP to undertake targeted larvicide applications (e.g., using VectoBac® GS granular Bti or Vectolex granular Bs) to protect human safety if the mean number of mosquito larvae is less than the threshold that is established in consultation with CT MMP. At a minimum, the threshold will be 5 or more larvae/dip with more than 50 percent of the dips (minimum of 3 dips/site) containing larvae per standard long-handled plastic cup dipper (8 oz.) taken on the same day within each source pool across the 5-acre site. This criterion is subject to change depending on the results of future coordination with the CT MMP. Mosquito vector populations below this level will not be treated. The CT MMP will coordinate with the refuge manager prior to surveillance, monitoring, and control activities occur on the refuge.

Variations in annual SUP restrictions may be necessary to accommodate wildlife breeding, roosting, and feeding activity, endangered species, administrative needs, public use management, research, or monitoring protocols. Other conflicts that may arise will be incorporated into the annual SUP to ensure there are no significant adverse impacts to refuge wildlife and habitats.

The CT MMP is required to provide the refuge manager with an annual quantitative summary of refuge mosquito monitoring and surveillance results, control activities on the refuge (e.g., type of pesticides applied, amount of pesticides applied, locations of application, method of application), and regional disease surveillance. All surveillance and control activities would be spatially referenced as technologies develop at CT MMP (e.g., use of global positioning satellites (GPS) and geographic information systems (GIS). Comparisons of mosquito management within and among years should be presented to permit analysis of patterns that may indicate success of habitat management efforts or suggest the need for a new management approach.

(e) Why is this use being proposed? The use is proposed to minimize health risks to humans and wildlife from mosquito-borne disease.

The Great Meadows Unit at the refuge lies within the jurisdiction of the CT MMP, which is an interagency state working group led by the CT DEEP and includes the Department of Public Health (DPH), the Connecticut Agricultural Experiment Station (CAES), the Department of Agriculture (DoAg), the University of Connecticut Department of Pathobiology and Veterinary Science (UConn), and the Connecticut Association of Directors of Health. Public Act 97-289, “An Act Concerning Mosquito Control and Aerial Application of Pesticides,” (CGS Sec 22a-45b) created the CT MMP and allows the Commissioner of Connecticut Energy and Environmental Protection or his/her agent to:

• adopt regulations or make orders regarding elimination of mosquitoes and their breeding

places; • enter land to determine whether mosquitoes breed there; and • survey, drain, fill, treat, excavate, construct or make other permanent alterations to eliminate mosquito breeding. Under the act, if such activities involve private property, the commissioner must have the consent of the owner.

This Act authorizes the necessary measures to abate any pest-borne threat, including prevention and remedial measures, and allows for the application of broad spectrum chemical pesticides to address an imminent peril to the public health, safety, or welfare posed by pests.

The original focus of the CT MMP was to monitor the threat of EEE, a potentially deadly disease. EEE is a virus that is present in nature and is cycled in the wild bird population by certain species of bird-feeding mosquitoes. The virus has no effect on wild birds; however, it can be fatal to humans, horses, and commercial exotic fowl (e.g., pheasants, emus). In Connecticut, outbreaks of EEE have occurred sporadically among horses and domestic pheasants since 1938.

In 1999, West Nile virus (WNV) was discovered in New York, New Jersey and Connecticut. The outbreak, in which seven humans and six horses died in New York and hundreds of birds died within the three states, was the first documentation of this virus in the Western Hemisphere. Unlike EEE, WNV is new to the Americas and native birds had not developed a natural immunity to this virus. Hence, a large proportion of the birds that are bitten by WNV-infected mosquitoes die. Historically, sporadic outbreaks of WNV have occurred in parts of Africa and Eurasia since 1937. The virus is similar to the virus that causes St. Louis encephalitis (SLE) and causes similar symptoms in humans. Although WNV causes fatal illness in a smaller proportion of cases than EEE, its greater potential to cause large outbreaks makes it an important health concern. Of the 114 WNV-associated illnesses reported to the DPH from 2000 to 2013, there were three human deaths associated with meningitis or encephalitis in patients over 80 years of age (http://www.ct.gov/dph/lib/dph/infectious_diseases/ctepinews/vol34no2.pdf; accessed January 2015).

Mosquito population monitoring is necessary to detect changes that indicate increased human health risks. In addition, surveillance for incidence of mosquito-borne disease by testing wildlife, especially birds, and adult mosquitoes for pathogens is needed to help characterize the level of health risk. As a result of the on-going human surveillance efforts, regional and temporal patterns of human illness and virus isolations from mosquitoes have been detected and help focus the public health response (http://www.ct.gov/dph/lib/dph/infectious_diseases/ctepinews/vol34no2.pdf; accessed January 2015). From 2000-2013, 93 percent of people with WNV infections acquired in-state were residents of urban and suburban towns with dense human population in three of Connecticut’s eight counties (Fairfield, New Haven, and Hartford counties). Based on the dates of onset of illness and typical incubation period, the risk for acquiring WNV infection is generally highest from early-August to early-September. These findings are also supported by mosquito surveillance data that provides early warning of regional presence of WNV infected mosquitoes, and detailed information for risk assessment. The goal of early mosquito larvae monitoring is rapidly detection of relative and absolute changes in population size that can indicate an increased short-term risk to human, wildlife, or domestic animal health.

As part of the statewide CT MMP, the CT DEEP has previously been allowed to monitor and control larval mosquito populations on the refuge, at both the Salt Meadow Unit in Westbrook and the Great Meadows Unit in Stratford. In the early 1990s, CT DEEP performed Open Marsh Water Management in the mosquito-producing areas of the Salt Meadow Unit, essentially eliminating salt

marsh mosquito-producing sites there. Then in 2001, CT DEEP assisted in the restoration of 40 acres of tidal wetlands as part of the Stratford Development Company mitigation project. In doing so, many of the mosquito-producing sites that were once part of CT DEEP’s larviciding program were also eliminated. However, there are several areas at Great Meadows that were not restored and still produce prolific numbers of mosquitoes after heavy rainfall or lunar high tidal events. The cumulative size of these sites is approximately 5 acres. There have been discussions for a few years now between the Service, CT DEEP, and NOAA/NMFS regarding additional tidal wetland restoration on the refuge in completion with the Lordship/Raymark Restoration Plan/Environmental Assessment. Like the work done in 2001, once this restoration is completed, salt marsh mosquito-producing areas on the refuge should be eliminated. In the interim, these sites continue to produce mosquitoes.

Several primary and bridge vector mosquito species associated with EEE and WNV transmission to humans have been detected on the refuge from CT MMP surveillance (from 2003 through 2007; table 2) and at nearby trapping sites outside of the refuge. Some of the mosquitoes detected are bridge vectors, meaning these species feed on birds and other animals, thereby enhancing the risk of disease transmission to people.

Table 2. Arbovirus Mosquito Vectors Potentially Found on Stewart B. McKinney NWR

Mosquito Vector EEEV WNV Vector Flight Range Known to Occur on Refuge Culiseta morsitans X Birds Coquillettidia perturbans X Bridge X Bridge 5 km Ochlerotatus canadensis X Bridge X Bridge 2 km Aedes vexans X Bridge X Bridge >25 km Culex pipiens X Bridge X Birds 2 km Yes Culex restuans X Bridge X Birds 2 km Yes Culex salinarius X Bridge X Bridge 10 km Yes Ochlerotatus excrusians Ochlerotatus sollicitans X Bridge X Bridge >25 km Yes (Formerly Aedes Ochlerotatus cantator X Bridge X Bridge >10 km Yes (Formerly Aedes contator) Ochlerotatus X Bridge Yes taeniorhynchus Ochlerotatus triseriatus X Bridge X Bridge 0.2 km

AVAILABILITY OF RESOURCES: The CT MMP will conduct monitoring and control, coordinated with the refuge manager on an annual basis through the issuance of a SUP. Existing funds are available to support the refuge manager and other staff in coordinating this use (table 3). As funding becomes available, refuge staff will take an active and, in most cases, a lead role in planning and implementing tidal circulation enhancement and wetland restoration projects aimed at improving wildlife habitat while reducing mosquito production on the Great Meadows Unit at Stewart B. McKinney NWR. Developing a mosquito management plan for the refuge will be a one-time effort that is likely to take 0.20 of a

full-time employee (FTE). A notice of intent would need to be submitted to the Environmental Protection Agency for the use of pesticides in the salt marsh, and it would be the responsibility of the CT MMP to draft a notice of intent and either acquire the permit, or provide all the information needed so the Service can obtain the permit. This will be listed as a condition for issuing a SUP for mosquito control.

Table 3. Staffing needs to conduct mosquito management on the Great Meadows Unit at Stewart B. McKinney NWR

Staff Member Involvement FTE Cost Refuge manager General oversight 0.04 $3,200 Wildlife biologist Field visits, mosquito management plan review and 0.07 $4,200 implementation; preparation of pesticide use proposal (PUP), special use permit (SUP), and pesticide use report; and oversight of mosquito-borne disease monitoring, vector control activities. Involvement in coordination and oversight of mosquito monitoring and control activities.

Total FTES and Staffing 0.11 FTE $7,400

ANTICIPATED IMPACTS OF THE USE: Direct impacts of monitoring and control include temporary disturbance to habitat and possible direct effects to non-target wildlife. Areas of vegetation may be crushed underfoot, with impacts ranging from temporary in nature to loss of habitat over time. Invasive weeds may be introduced or spread by foot. Indirect effects associated with mosquito control include reducing mosquito populations and other non-target species that serve as the base of food chains for wildlife species.

Impacts to birds as a result of physical access (trampling of vegetation, nests) for mosquito management could occur, but are unlikely, as these actions would not significantly affect bird populations on the Great Meadows Unit at the refuge given the small size and limited bird habitat that the areas receiving mosquito management provide.

Chemical Treatment Effects on Target Mosquito Populations The use of mosquito larvicides generally is considered preferable to the use of adulticides because larvicides prevent the appearance of the blood feeding adults; larvicides can provide up to a month of control, rather than the few hours provided by fogging with adulticides; the commonly used larvicides are less toxic than the adulticides and the application method greatly reduces human exposure; and larvicides generally are applied to smaller areas than are adulticides.

A natural soil bacterium, Bacillus thuringiensis var. israelensis (Bti) and Bacillus sphaericus (Bs), like other varieties of Bacillus sp., is a stomach poison that must be ingested by the larval form of the insect in order to be effective. Bti is an USEPA toxicity class III general use pesticide and is practically non-toxic to animals (Extoxnet 1996). Bti is specific to certain primitive dipterans, especially mosquitoes, black flies, and some chironomid species (Boisvert and Boisvert 2000), and is not known to be directly toxic to non-dipteran insects; there are no toxic inert ingredients included in Bti products (Extoxnet 1996). Bti produces protein endotoxins, activated in the alkaline mid-gut

of target insect species that bind to protein specific receptors of dipteran larvae species, resulting in mortality. Bti must be ingested by the target insect to be effective and is most effective on larval salt marsh mosquito instar stages 1 and 2; it is considerably less effective against instar stages 3 and 4. Bti has no effect on pupae or adult mosquitoes. Because Bs is a more recently developed larvicide than Bti, there are fewer studies that have examined the non-target effects of this pesticide. The data available, however, indicate a high degree of specificity of Bs for mosquitoes, with no demonstrated toxicity to chironomid larvae at any mosquito control application rate (Mulla et al. 1984; Ali and Nayar 1986; Lacey and Mulla 1990).

Pupacides (surface films) are applied to mosquito breeding sites to kill mosquito larvae and pupae. The products create a barrier to the air-water interface and suffocate insects, which require at least periodic contact with the water surface in order to obtain oxygen. Surface films are alcohol based and produce a monomolecular film over the water surface. Monomolecular films are potentially lethal to any aquatic insect that lives on the water surface and requires periodic contact with the air- water interface to obtain oxygen. The film interferes with larval orientation at the air-water interface and/or increases wetting tracheal structures, thus suffocating the organism. As the film spreads over the water surface, larvae tend to concentrate, which may increase mortality from crowding stress (Dale and Hulsman, 1990).

Adulticides appear to effectively control adult mosquito populations and the spread of mosquito-borne diseases such as WNV (Carney et al. 2008), but only for a brief time, and are therefore only recommended during a disease event to break the disease transmission cycle (http://www.townofsilvercity.org/vector/ scienceofvector.html). Adulticides kill only mosquitoes that contact insecticide droplets. The fog soon dissipates. Although the local mosquito population is reduced for a few days, fogging does not prevent mosquitoes from re-entering the sprayed area. Adulticides will only be considered in the case of a declared public health emergency. Focused timing and location of adulticide application to control mosquito disease vector source populations is essential for effectiveness (http://wildpro.twycrosszoo.org/s/00man/WNVOverviews/wnvindtech/wnvcontrolaerialadulticides.htm).

The Ecotoxnet database (http://extoxnet.orst.edu/) includes the following summary of how Pyrethrin/Pyrethroids act as insecticides: “Human-made Pyrethrin/Pyrethroids are based on natural pyrethrins in chrysanthemums, which are a neurotoxic chemical to insects. Pyrethrin/Pyrethroids act by inhibiting the nervous system of insects. This occurs at the sodium ion channels in the nerve cell membrane. Some type II Pyrethrin/Pyrethroids also affect the action of a neurotransmitter called GABA. Pesticide products containing pyrethrins usually contain a synergist (such as piperonyl butoxide). Synergists work by restricting an enzyme that insects use to detoxify the pyrethrins. A synergist allows the insecticide to be more effective. These products are dissolved in petroleum-based products.”

Pesticide Toxicity and Other Effects to Non-target Organisms The few small refuge sites that would receive pesticide application for the purpose of mosquito management typically provide limited habitat for native wildlife and plants. These areas are mostly shallow swales within the intertidal marsh plain (4 to 6 feet wide) that hold water for extended periods (e.g., following high tides); the area lacks tidal channels that permit drainage. These characteristics result in poor tidal hydrology and, in turn, lower biotic productivity for a variety of plant and wildlife species relative to other refuge areas with better tidal flushing.

Giving full consideration to the protection and integrity of non-target organisms and communities, the greatest concerns the Service has with chronic mosquito larvicide control chemical use are the

subsequent degradation of biological integrity and diversity, and disruption of vital food webs. Aquatic invertebrates play important roles in wetland ecology. They aid in the breakdown of fresh and salt marsh-derived organic matter and provide important food resources for different life stages of fish, breeding and migrating birds, and other wildlife. As such, they are critically important and directly linked to the future conservation and management of refuge-specific resources of concern listed in Comprehensive Conservation Planning (CCP) goals and habitat objectives.

Impacts to birds, mammals, reptiles, or amphibian may occur as a result of ground access. However, bird and mammal impacts are considered limited because areas that need mosquito management are small in size, and provide only limited habitat. The use of pesticides for the purpose of mosquito management may directly or indirectly affect resident and migratory bird, mammal, reptile or amphibian populations of the refuge. Direct effects may occur from direct contact with the pesticides. Indirect effects are related to the potential reduction in the invertebrate food supply. Pesticide effects on reptiles and amphibians may occur through reductions in insects that serve as a food source (Hoffman et al. 2008), through direct individual effects from pesticide application, or from trampling of individuals or habitat. Birds are often used as a surrogate for effects on reptiles, and fish as a surrogate for amphibians (Hoffman et al. 2008). Bti and Bs have practically no acute or chronic toxicity to mammals, birds, fish, or vascular plants (USEPA 1998, USEPA 2014).

Migratory birds that depend on invertebrate food resources may not be mobile enough to seek alternative feeding sites, post-treatment, particularly during the breeding season. Precocial young seek food items on their own. Since they are flightless, food items must be available within a relatively small home area. Reduction of invertebrate food resources within even a small geographic area may be detrimental to breeding wetland birds and precocial young.

The young of altricial birds are solely dependent upon the parents for food as they are relatively helpless and restricted to a discrete nest site during the first few weeks of life. When invertebrate foods are scarce, parents may have to make more extended feeding forays and be less able to provide sufficient nutrition to all offspring, potentially resulting in increased chick mortality. Adults making extended flights into less familiar territory may be more likely to suffer predation or to experience inter- or intra-specific competition. Young birds that are subjected to extended periods in the nest without parental attention may be more likely to suffer predation or weather-related stress.

The use of larvicides and pupacides for the purpose of mosquito management is not likely to directly affect native mammal populations of the refuge. Adverse effects on mammals from Bti, methoprene, and Agnique (monomolecular film) are not expected when applied according to the label instructions. Extensive acute toxicity studies indicated that Bti is virtually innocuous to mammals (Siegel and Shadduck 1992). These studies exposed a variety of mammalian species to Bti at moderate to high doses and no pathological symptoms, disease, or mortality were observed. Impacts to the mammalian community as a result of reduced invertebrate populations are not expected because most mammal species that inhabit refuge wetlands are herbivores and invertebrates are not a primary component of their diet. Insectivorous shrews may be reduced over the short-term, post-treatment, if they experience reduced arthropod food availability. Negative effects on fish populations are not expected from proposed larvicides and pupacides.

Using larvicides can adversely affect non-target insects, especially non-biting midges (Chironominae). Bti concentration is important with regard to impacts on non-target organisms such as ecologically important non-biting midge larvae. Chironomid larvae are often the most abundant aquatic insects in freshwater, brackish, and salt marsh wetland environments and represent a major component in food webs for many wetland-dependent wildlife species (Miller 1987, Euliss et al.

1991, Helmers 1992, Skagen and Oman 1996, MacKenzie 2005). Chironomids also frequently make up the largest proportion of wetland invertebrate biomass (Eldridge 1992, Rehfisch 1994, Davis and Smith 1998, MacKenzie 2005).

The effect on local populations of invertebrate species over time with periodic and continued use of Bti is unknown but the potential for negative effects is a possibility. Host range and effect on non-target organisms indicates that Bti is relatively specific to the Nematocera suborder of Diptera, in particular filter-feeding mosquitoes (Culicidae) and blackflies (Simuliidae) (Glare and O’Callaghan 1998). Bti is pathogenic to some species of midges (Chironomidae) and Tipulidae, although to a lesser extent than to mosquitoes and biting flies; it is not reported to affect a large number of other invertebrate species (Glare and O’Callaghan 1998). Other factors, such as temperature, water depth, aquatic vegetation, and suspended organic matter, may act to reduce its toxicity to chironomids in the environment (Charbonneau et al. 1994; Merritt et al. 1989, Lacey and Merritt 2004). Negative impacts on chironomid density and biomass could have deleterious effects on wetland and wildlife food webs and could lower biodiversity. The effects of a single Bti application are difficult to predict because of documented differences in toxicity based on formulation, potency, application rate, and timing. Published studies (Hershey et al. 1998, Niemi et al. 1999) have examined the long-term, non-target effects of Bti. In Minnesota, 27 wetlands were sampled for macroinvertebrates over a 6-year period with no effects observed on the bird community (Niemi et al. 1999). In judging the potential for adverse ecological effects of Bti applications, one should consider the non-target aquatic organisms of concern that would be impacted from the potential loss of both mosquito and chironomid larvae.

The extent to which the use of Bti or Bs will limit the food resources for individual birds or local avian populations is unknown. Though often discounted as inefficient pollinators, some researchers have suggested that the efficiency of pollinating flies (dipterans), mosquitoes (dipterans), and midges can exceed that of bees (http://eol.org/pages/421/entries/24921263/details#relevance_to_humans_and_ ecosystems). Further, dipterans appear to be crucial for the pollination of certain flowers in some habitats.

Monomolecular films are not known to cause direct chronic or acute toxicological effects to birds, but are potentially lethal to any aquatic insect that lives on the water surface or requires periodic contact with the air- water interface to obtain oxygen; this may result in a negative impact to the avian food base, e.g., chironomid invertebrates (USFWS 2005). The film interferes with larval orientation at the air-water interface or increases wetting of tracheal structures, suffocating the organism. As the film spreads over the water surface, larvae tend to concentrate, which may increase mortality from crowding stress (Dale and Hulsman 1990).

Pyrethroid insecticides are subject for review as potential developmental neurotoxicants because of their mode of action on voltage-sensitive sodium channels (Lu et al. 2006). Permethrin, the most widely used pyrethroid insecticide, is suspected to be an endocrine-disrupting chemical and was classified as a potential carcinogen at high exposure levels (USEPA 2006). Pyrethrin/Pyrethroids may also have a suppressive effect on the immune system and may cause lymph node and spleen damage. Pyrethrin/Pyrethroids are reported to degrade rapidly in the environment and to be broken down to nontoxic products. However, Tyler et al. (2000) and Hong Sun et al. (2007) argue that products of the metabolism of permethrin are potentially far more potent as endocrine disruptors than the parent compound because of their ability to interact with steroid hormone receptors. Pyrethrines/Pyrethroids have a slight toxicity to bird species (National Pesticide Information Center 1998, Extoxnet 1994). Non-target effects to birds from pyrethrin application may also occur as a result of a reduced food base (e.g., chironomid invertebrates) if non-target invertebrate populations

are significantly reduced.

The application of adulticides has the potential to adversely affect fish and aquatic invertebrate populations. Pyrethrines/Pyrethroids are considered highly toxic to fish and invertebrates (USEPA 2006).

Because Pyrethrines/Pyrethroids are broad-spectrum insecticides, they are potentially lethal to most insects. All adulticides are very highly toxic to aquatic invertebrates in concentrations as low as one part per billion (Milam et al. 2000). Pyrethrines/Pyrethroids are known to cause acute toxicological effects to benthic invertebrates at rates used for mosquito abatement (USEPA 2006). Because most adulticides can be applied over or near water when used for mosquito control, risks to aquatic invertebrates from direct deposition and runoff of the pesticides exist.

The pyrethroid insecticides are extremely toxic to fish, with 96-hour LC50 values generally below 10 ug/l. Corresponding LD50 values in mammals and birds are in the range of several hundred to several thousand mg/ kg. Fish sensitivity to the Pyrethrin/Pyrethroids may be explained by their relatively slow metabolism and elimination of these compounds. The half-lives for elimination of several Pyrethrin/Pyrethroids by trout are all greater than 48 hours, while elimination half-lives for birds and mammals range from 6 to 12 hours. Generally, the lethality of Pyrethrin/Pyrethroids to fish increases with increasing octanol/water partition coefficients. The pyrethroid resmethrin is slightly toxic to birds and highly toxic to fish and to bees. Its LD50 in California quail was greater than 2,000 mg/kg; the LC50 in mosquito fish is 0.007 ppm. The LC50 for resmethrin synergized with piperonyl butoxide in red swamp crawfish, Procambarus clarkii, is 0.00082 ppm. The LC50 in bluegill sunfish is 0.75 to 2.6 ug/l, and 0.28 to 2.4 ug/l in rainbow trout. DeMicco et al. (2010) found a dose-dependent increase in zebrafish embryo mortality and pericardial edema, which was consistent with mammalian studies that demonstrated slight teratogenesis at high doses. Resmethrin is highly toxic to bees, with an LD50 of 0.063 ug/bee. Adulticides (Pyrethrines/Pyrethroids) may adversely affect amphibians such as tadpoles that occur within seasonal freshwater wetlands of the refuge (Gunasekara 2005).

De Guise et al. (2005) studied a die-off of lobsters following mosquito spraying with resmethrin; they found that adult lobsters are no more sensitive than other aquatic species to the lethal effects, but are very sensitive to immune and endocrine endpoints tested (sublethal effects). Modulation in immune functions could result in increased susceptibility to infectious agents, contributing to mass mortality with sufficient exposure. Weston et al. (2005) examined toxicity of run-off sediments to an amphipod Hyalella azteca in creeks draining a Roseville, California, single-family subdivision. Nearly all creek sediments collected caused toxicity in laboratory exposures, and about half the samples caused nearly complete mortality. The pyrethroid bifenthrin was implicated as the primary cause of the toxicity, with additional contributions to toxicity from the pyrethroids cyfluthrin and cypermethrin originating from residential (structural) pest control by professional applicators or homeowner use of insecticides, particularly lawn care products.

The small scale and low frequency in past use of adulticides suggests that any future adulticide use on the refuge is unlikely to cause significant adverse effects to fish and invertebrate populations. Application would only occur in swales and not to channels, sloughs, or other open water areas. Application would only occur during low tides to avoid potential impacts to fish that may move into the tidal marsh plain during higher high or extreme tides. Oral exposure of mammals to Pyrethrines/Pyrethroids could occur through consumption of plants or plant parts that have been sprayed. A terrestrial exposure model showed no acute or chronic risks to mammal or bird species (USEPA 2006).

The Service recognizes that spray drift could enter the refuge from neighboring communities. The refuge has no jurisdiction over mosquito control on lands outside the refuge boundary; therefore, no SUP is required for off-refuge mosquito management. Since the State employs best management practice and follows the USEPA-approved label, the Service expects impacts to refuge resources to be minimal.

Refuge habitat management actions that increase biological integrity, diversity, and environmental health (BIDEH) and avian diversity have the potential to provide a buffer against future disease outbreaks. Recent infectious disease models illustrate a suite of mechanisms that can lower incidence of disease in areas of higher disease host-diversity (defined as the dilution effect). These models are particularly applicable to human zoonosis (i.e., infectious diseases of wildlife or domestic animals that spill over into human populations) (Keesing et al. 2006, Krasnov et al. 2007, Ostfeld and Kessing 2000 (a&b)), such as avian influenza, anthrax, Lyme disease, and West Nile virus.

PUBLIC REVIEW AND COMMENT: A news release announcing the availability of the draft compatibility determination (CD) for a 15- day public review and comment period was issued the following media outlets on [DATE]: [MEDIA CONTACTS]. The draft CD was distributed to representatives of the [groups, organizations, agencies].

A copy of the draft CD was made available for public review and comment at these locations: • Refuge Headquarters: [ADDRESS]

• Internet: http://www.fws.gov/refuge/Stewart_B_McKinney

DETERMINATION (CHECK ONE BELOW):

Use is not compatible X Use is compatible, with the following stipulations

STIPULATIONS NECESSARY TO ENSURE COMPATIBILITY: The following stipulations are required to ensure compatibility: • The CT MMP must apply for and receive a special use permit (SUP) annually from the refuge manager prior to conducting any mosquito and mosquito-borne disease surveillance and monitoring activities.

• The CT MMP will notify the refuge manager prior to monitoring and conducting disease surveillance. All personnel entering the wetlands will be oriented at the beginning of the surveillance period or escorted by refuge staff to avoid disturbance to endangered, threatened, or other sensitive species on the refuge.

• The CT MMP will be responsible for monitoring disease activity in reservoir hosts for pathogens or antibodies, and collecting adult mosquito samples in same-genus pools for virus or any other monitoring required to substantiate a high-risk disease situation on or near the refuge.

• The CT MMP will assume all monetary costs and perform all activities associated with mosquito monitoring, disease surveillance, and treatment. Service personnel may accompany CT MMP personnel to examine exact locations of heavy mosquito breeding problems to ascertain the presence of non-targets or mosquito predator species in these areas.

• The CT MMP will limit the number of travel pathways used for mosquito management within the marsh.

• Caged sentinel chickens may not be used for reservoir host surveillance due to the risk of spreading disease to wild birds.

• The CT MMP will remove equipment and refuse resulting from operations on refuge lands daily, and will promptly repair all damage to government property that may result.

• All decisions for chemical interventions to control mosquitoes will be made by the refuge manager and will be based on meeting or exceeding predetermined mosquito abundance and disease thresholds.

• Current mosquito population data is necessary before mosquito larvicide treatments may be applied on the refuge.

• Only approved larvicides may be applied on refuge salt marshes within the prescribed area as identified in the special use permit.

• The refuge manager will be contacted at least 24 hours in advance of each larvicidal application.

• The CT MMP must provide a copy of the Clean Water Act NPDES permit from the Environmental Protection Agency prior to conducting any chemical treatment.

• Application of chemical mosquito control measures will be conducted in accordance with approved pesticide use proposals (PUP).

• Insecticide applications will avoid areas known to contain butterfly and moth host-plants in order to conserve and protect rare or specialist insect pollinators and also ensure that adequately buffered habitat around host plants or refugia is available during and after insecticide spraying.

• Application of pesticides will be in discrete, mosquito-producing areas of on the Great Meadows Unit at the refuge and at the lowest possible dilution rate (ultra-low volume) required for effectiveness.

• The CT MMP will minimize the use of pesticides on the Great Meadows Unit at the refuge, and continually investigate formulations and compounds that are least damaging to fish and wildlife populations.

• The CT MMP must provide the refuge manager with monitoring and disease surveillance data demonstrating that action thresholds have been reached or exceeded before pupacides are applied. Refuge manager approval must be obtained prior to CT MMP staff elevating to the next action or response threshold.

• Only the refuge manager, in consultation with the CT MMP and public health officials, may authorize application of mosquito adulticide and only when there is evidence of refuge-based mosquitoes contributing to a declared public health emergency.

• Immediately after any pesticide application, the CT MMP will monitor mosquito vector populations to assess the effectiveness of all pesticide treatments.

• Treatment in populated areas off-refuge will be considered first.

• General mosquito control will not be allowed during high tide events in order to avoid impacts to tidal marsh species. Unless permitted by the refuge manager, pesticide application should not occur within 100 feet of natural sloughs and channels.

• A final report of all monitoring and control activities conducted on the Great Meadows Unit at the refuge must be provided to the refuge manager before the end of the calendar year.

• The CT MMP will meet with the refuge manager during the first quarter of each calendar year as a condition of the SUP renewal for the upcoming year. Prior to that meeting, the CT MMP will review the previous year’s pesticide proposals and submit to the refuge manager any changes in the pesticides or formulations of pesticides they expect to use in the upcoming year.

JUSTIFICATION: Mosquitoes are a natural component of tidal wetlands but can pose a significant potential threat to human health when refuge wetlands are within the known mosquito flight ranges of populated areas and refuge mosquitoes have been demonstrated to be infected with arboviruses. WNV and EEE have been of particular concern across the United States and Long Island Sound. Mosquito species known as vectors of these diseases occur on the Great Meadows Unit at the refuge.

The staffs of Stewart B. McKinney NWR and the CT MMP advocate an integrated approach to mosquito management that includes a range of tools to improve habitat conditions for estuarine

wildlife while reducing threats to public health from mosquito species capable of transmitting disease to humans. With the continued existence of WNV and EEE and the potential for spread of other mosquito-borne disease, pressure is increasing to manage mosquito populations that occur on lands of the National Wildlife Refuge System, especially in populated areas near the Long Island Sound. Understanding the actual risk of refuge-based mosquitoes to the spread of WNV and EEE is an important part of managing a mosquito control program on the Great Meadows Unit at the refuge.

The use of larvicides and other pesticides, if necessary, will receive periodic compatibility review if future studies bring more information to light on the ecological impacts of mosquito control. In addition, new chemicals that may come to market in the future may be evaluated for potential use on the Great Meadows Unit at Stewart B. McKinney NWR.

The stipulations above address the Service’s laws and Refuge System policies to maintain, enhance, and restore biological integrity, diversity, and environmental health, manage an IPM program, and protect the public from mosquito-borne health threats.

This activity will not materially interfere with or detract from the mission of the National Wildlife Refuge System or the purpose for which the refuge was established.

SIGNATURE: Refuge Manager: ______

(Signature) (Date)

CONCURRENCE: Regional Chief: ______(Signature) (Date)

MANDATORY 10 YEAR RE-EVALUATION DATE: ______

LITERATURE CITED:

Ali, A. and J. K. Nayar. 1986. Efficacy of Bacillus sphaericus Neide against larval mosquitoes (Diptera: Culicidae) and midges (Diptera: Chironomidae) in the laboratory. Florida Entomologist 69: 685-690.

Carney, R.M., S. Husted, C. Jean, C. Glaser, and V. Kramer. 2008. Efficacy of Aerial Spraying of Mosquito Adulticide in Reducing Incidence of West Nile Virus, California, 2005. Emerging Infectious Diseases 14(5).

Centers for Disease Control (CDC). 2010. West Nile Virus Home Page. Available at http://www.cdc.gov/ncidod/dvbid/westnile/index.htm

Charbonneau, C.S., R.D. Drobney, and C.F. Rabeni. 1994. Effects of Bacillus thuringiensis var. israelensis on nontarget benthic organisms in a lentic habitat and factors affecting the efficacy of the larvicide. Environmental Toxicology and Chemistry 13: 267-279.

Connecticut Department of Energy and Environmental Protection (CT DEEP). 2012. Connecticut West Nile Virus Surveillance and Response Plan, 2012. Accessed January 2015 at: http://www.ct.gov/mosquito/lib/mosquito/publications/wnvplan.pdf.

Connecticut General Statutes (CGS). 2013. Title 22a – Environmental Protection. Accessed January 2015 at: http://www.cga.ct.gov/2013/pub/title_22a.htm.

Dale, P.E.R. and K. Hulsman. 1990. A critical review of salt marsh management methods for mosquito control. Review in Aquatic Sciences 3: 281-311.

Davis, C. A. and L. M. Smith. 1998. Ecology and management of migrant shorebirds in the playa lakes region of Texas. Wildlife Monographs 140: 1–45.

De Guise, S., J. Maratea, E.S. Chang, and C. Perkins. 2005. Resmethrin immunotoxicity and endocrine disrupting effects in the American lobster (Homarus americanus) upon experimental exposure. Journal of Shellfish Research 24(3): 781–786.

DeMicco, A., K.R. Cooper, J.R. Richardson, and L.A. White. 2010. Developmental Neurotoxicity of Pyrethroid Insecticides in Zebrafish Embryos. Toxicological Sciences 113(1): 177–186.

Eldridge, J. 1992. Management of habitat for breeding and migrating shorebirds in the Midwest. Chapter 13.2.14 In U.S. Fish and Wildlife Service Waterfowl Management Handbook. Washington, D.C.

Euliss, N. H., Jr., R. L. Jarvis, and D. S. Gilmer. 1991. Standing crops and ecology of aquatic invertebrates in agricultural drainwater ponds in California. Wetlands 11: 179-190.

Extension Toxicology Network (Extoxnet). 1994. Pyrethrins and Pyrethroids. Available at http://extoxnet.orst. edu/pips/pyrethri.htm.

Extension Toxicology Network (Extoxnet). 1996. Bacillus thuringiensis. Pesticide Information Profile.

Extension Toxicology Network. Available at http://extoxnet.orst.edu/pips/bacillus.htm, Accessed May 6, 2012.

Glare, T.R. and M. O’Callaghan. 1998. Environmental and health impacts of Bacillus thuringiensis israelensis. Report for New Zealand Ministry of Health. 58 p.

Gunasekara, A.S. 2005. Environmental Fate of Pyrethrins. California Department of Pesticide Regulation, Environmental Monitoring Branch, Sacramento, California. 19 pp.

Helmers, D.L. 1992. Shorebird Management Manual. Western Hemisphere Shorebird Reserve Network, Manomet, 58 pp.

Hershey, A.E., A.R. Lima, G.J. Niemi, and R.R. Regal. 1998. Effects of Bacillus thuringiensis israelensis (Bti) and methoprene on nontarget invertebrates in Minnesota wetlands. Ecological Applications8: 41-60.

Hoffmann, M., J.L. Melendez, and M.A. Mohammed. 2008. Risk of permethrin use to the federally threatened California red-legged frog and bay checkerspot butterf ly, and the federally endangered California clapper rail, salt marsh harvest mouse, and San Francisco garter snake. Pesticide Effects Determination.

Hong S., X.L. Xu, L.C. Xu, L. Song, X. Hong, J.F. Chen, L.B. Cui, X.R. Wang. 2007. Antiandrogenic activity of pyrethroid pesticides and their metabolite in reporter gene assay. Chemosphere 66: 474–479.

Keesing, F., R.D. Holt, and R.S. Ostfeld. 2006. Effects of species diversity on disease risk. Ecology Letters 9(4) 485-498.

Krasnov, B.R., M. Stanko, and S. Morand. 2007. Host community structure and infestation by ixodid ticks: repeatability, dilution eVect and ecological specialization. Oecologia 154: 185–194.

Lacey, L. A. and M. S. Mulla 1990. Safety of Bacillus thuringiensis ssp. israelensis and Bacillus sphaericus to nontarget organisms in the aquatic environment, p. 169-188. In M. Laird, L.Lacey, and E. Davidson [eds.], Safety of Microbial Insecticides. CRC Press. Baco Raton, FL.

Lacey, L. A. and R.W. Merritt. 2004. The safety of bacterial microbial agents used for black f ly and mosquito control in aquatic environmentsKluwer Academic Publishers Netherlands. Appears in: Environmental Impacts of Microbial Insecticides: Need and methods for Risk Assessment.

LoGiudice,K., R.S. Ostfeld, K.A. Schmidt, and F. Keesing. 2003. The Ecology of Infectious Disease: Effects of Host Diversity and Community Composition on Lyme Disease Risk. Proceedings of the National Academy of Sciences 100(2): 567-571.

Lu, C., D.B. Barr, M. Pearson, S. Bartell, and R. Bravo. 2006. A Longitudinal Approach to Assessing Urban and Suburban Children’s Exposure to Pyrethroid Pesticides. Environmental Health Perspectives Vol. 114 (9): 1419-1423.

MacKenzie, R.A. 2005. Spatial and temporal patterns in insect emergence from a southern Maine salt marsh. American Midland Naturalist 153: 257-269.

Meredith, W.H., D.E. Saveikis, and C.J. Stachecki. 1985. Guidelines for open marsh water management in Delaware’s salt marshes – Objectives, system designs, and installation procedures. Wetlands 5:119-133.

Merritt, R.W., E.D. Walker, M.A. Wilzbach, K.W. Cummins, and W.T. Morgan. 1989. A broad evaluation of Bti for black f ly (Diptera: Simuliidae) control in a Michigan river: Efficacy, carry and nontarget effects on invertebrates and fish. Journal of the American Mosquito Control Association 5: 397-415.

Milam, C.D., J.L. Farris, and J.D. Wilhide. 2000. Evaluating Mosquito Control Pesticides for Effect on Target and Non-target Organisms. Archives of Environmental Contamination and Toxicology 39: 324-328.

Miller, M. R. 1987. Fall and winter foods of northern pintails in the Sacramento Valley, California. Journal of Wildlife Management 51: 403–412.

Moore, C.G., R.G. McLean, C.J. Mitchell, R.S. Nasci, T.F. Tsai, C.H. Calisher, A.A. Marfin, P.S. Moore, and D.J. Gubler. 1993. Guidelines for arbovirus surveillance programs in the United States. Centers for Disease Control, Ft. Collins, Colorado. 81 pp.

Mulla, M. S., H. A. Darwazeh, E. W. Davidson, H. T. Dulmage, and S. Singer. 1984. Larvicidal activity and field efficacy of Bacillus sphaericus strains against mosquito larvae and their safety to nontarget organisms. Mosquito News 44: 336-342.

National Pesticide Information Center. 1998. Pyrethrins & Pyrethroids. Available at http://npic.orst.edu/factsheets/pyrethrins.pdf.

Niemi, G.J., A.E. Hershey, L. Shannon, J.M. Hanowski, A. Lima, R.P. Axler, and R.R. Regal. 1999. Ecological effects of mosquito control on zooplankton, insects, and birds. Environmental Toxicology and Chemistry. 18(3): 549-559.

Ostfeld, R.S. and F. Keesing. 2000a. The function of biodiversity in the ecology of vector-borne zoonotic diseases. Canadian Journal of Zoology 78: 2061–2078.

Ostfeld, R.S. and F. Keesing. 2000b. Biodiversity and Disease Risk: The Case of Lyme Disease. Conservation Biology 14(3): 722-728.

Rehfisch, M.M. (1994) Man-made lagoons and how their attractiveness to waders might be increased by manipulating the biomass of an insect benthos. Journal of Applied Ecology 31: 383–401.

Siegel, Joel, P. and J. A. Shadduck. 1992. Mammalian safety of Bacillus thuringiensis israelensis and Bacillus sphaericus. Pp. 202-217 in de Barjac, Huguette and Donald J. Sutherland, eds.

Bacterial control of mosquitos and black flies: biochemistry, genetics, and applications of Bacillus thuringiensis israelensis and Bacillus sphaericus. Kluwer Academic.

Skagen, S.K. and H.D. Oman. 1996. Dietary flexibility of shorebirds in the western hemisphere. Canadian Field-Naturalist 110(3): 419-444.

Swaddle, J.P. and S.E. Calos. 2008. Increased Avian Diversity Is Associated with Lower Incidence of Human West Nile Infection: Observation of the Dilution Effect. PLoS ONE e2488 3(6): 1-8.

U.S. Environmental Protection Agency (USEPA). 1998. Re-registration eligibility document. Bacillus thuringiensis. Office of Prevention, Pesticides and Toxic Substances. EPA738-R-98-004.

U.S. Environmental Protection Agency (USEPA). 2014. Bacillus sphaericus: pesticide fact sheet. Environmental Protection Agency.

U.S. Environmental Protection Agency (USEPA). 2006. Permethrin Facts: Reregistration Eligibility Decision Fact Sheet. Available at http://www.epa.gov/oppsrrd1/REDs/factsheets/permethrin_ fs.htm accessed 05/07/2012.

U.S. Fish and Wildlife Service (USFWS). 2005. Interim Guidance for Mosquito Management on National Wildlife Refuges. 20 pp.

U.S. Fish and Wildlife Service (USFWS). 2017. Mosquito Management Plan and Environmental Assessment for the Great Meadows Unit at the Stewart B. McKinney National Wildlife Refuge.

Weston, D.P., R.W. Holmes, J. You, and J. Lydy. 2005. Aquatic toxicity due to residential use of pyrethroid insecticides. Environmental Science and Technology.

Wolfe, Roger. 2014 and 2015. Personal communication.

APPENDIX D. USFWS Pesticide Use Proposal – Example Form

U.S. Department of Interior Pesticide Use Proposal

U.S. Department of Interior Pesticide Use Proposal "tab" moves between fields PUP #: R1- CY Org Code Number R1- 2005 Refuge, Complex, Hatchery or Other Site Name:

County and State: County State

Contact Person and Phone: Person Phone Fax

Crop/Habitat of Treatment Site:

Location of Proposed Application (mgt. Unit or other unique ID), with map if available

Site Management Goal(s):

Need(s) for Treatment: Invasive species State or Federally listed noxious species Crop pest Native habitat restoration

Habitat improvement listed species protection/recovery public health protection wildlife health protection other Specify other

Target Pest(s) -- list all: Common Name & Binomial Nomenclature

Is there a monitoring plan for the target pest(s)? yes no

Proposed Action Threshold(s) Triggering Treatment:

Year of last approved IPM plan (attach if available):

Is this pesticide use part of your IPM plan? yes no

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U.S. Department of Interior Pesticide Use Proposal

Will non-chemical control methods be attempted? yes no If "no", explain rationale for pesticide use if different than previous descriptions:

Trade Name(s):

EPA Reg. Number(s):

Common or Chemical Name(s):

Manufacturer(s):

Please attach or provide URL for label(s) and MSDS(s):

Are the Pest(s) in this PUP Listed on the Label? Yes No

Is the Crop, Type of Vegetation, or Site Type Listed on the Label? Yes No

If the crop, type of vegetation, or site type is not listed, is there a current Section 24(c) or Section 18 exemption under which you are proposing to operate? Yes No N/A

Is use of the proposed pesticide part of any trial to compare different methods of treatment? Yes No

Is This labeled as a Restricted Use Pesticide (RUP)? Yes No

If a Restricted Use Pesticide: Certified Pesticide Applicator ID#: Company: Expiration date:

If a General Use Pesticide: Lead Pesticide Applicator Name: Company:

Is this a tank mix? Yes No

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U.S. Department of Interior Pesticide Use Proposal

Formulation: aqueous flowable aqueous suspension dust dry flowable Emulsifiable Concentrate

check one flowable microencapsulated granule solution Wettable Powder Other Specify other

Toxic Inert Ingredients Listed on MSDS:

Trade Names of Adjuvants (Drift Control Agents, Stickers, Surfactants, Oils):

Application Date(s):

Number of Applications:

Not to Exceed Limits on Label (lbs a.i./acre/season):

Product Application Rate(s) Per Acre Proposed: ------> pounds ounces fluid ounces check one pints quarts gallons other Specify other

Maximum Active Ingredient Rate Allowed on Label, if specified (lb ai/acre):

Application Method (check): Broadcast Directed-spray Backpack spray Cut-stump Frill Basal spray Injection Wick/wipe Ultra low volume (ULV) Chemigation other Specify other

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U.S. Department of Interior Pesticide Use Proposal

Application by (check as many as needed): Hand-Held Backpack Fogger Wet-blade mower Boom ATV Truck Boat Fixed-wing Helo other Specify other

Estimated Maximum Size of Treatment Area(s) (to nearest acre):

If spot treatment, Estimated Average Percent Cover To Be Treated (if not 100%):

Average Monthly Rainfall at Site During Proposed Application Period(s) (use range if multiple months): inches Soil Texture(s): clay silty clay sandy clay clay loam silty clay loam sandy clay loam silt silt loam loam sandy loam loamy sand sand gravel other Specify other

Organic Matter in Soil (if known); range if more than one site: %

Slope of Treatment Site: flat <3o <10o o >10

Soil pH, if known: If unknown pH < 7 pH ~ 7 pH > 7.5

Top Soil (to 3-ft Depth) during/ following treatment: Dry Moist Saturated Not predictable

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U.S. Department of Interior Pesticide Use Proposal

Shallowest Depth to Groundwater (check): <1 ft <5 ft < 10 ft < 100 ft > 100 ft Unknown

Distance to closest drinking water source (well or surface water intake): <0.25 mile <0.5 mile < 1 mile < 2 miles > 2 miles unknown

Closest Water to Treatment Site(s): N/A Pond Ditch Spring Drain Lake Canal Estuary Creek/stream/branch/run Ocean River Hatchery Wetland other Specify other

Nearest Distance of Treatment Site to Waterbody (check): < 25 ft < 50 ft < 100 ft < 150 ft < 300 ft < 400 ft > 400 ft unknown

Organisms which may occur at/near treatment site during or immediately after treatment (check): Sensitive plant species native lepidopterans native pollinating insects honeybees mussels crustaceans fish amphibians reptiles passerines shorebirds piscivorous birds waterfowl mammals other specify other

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U.S. Department of Interior Pesticide Use Proposal

If no written plan is available, describe other IPM methods used for the pests listed in this PUP: Describe sanitation, crop rotations, changes to resistant crop varieties, changes in timing, elimination of alternate host species, fallowing, cover crops, tillage, open-water marsh management, moisture/water manipulations, burning, mechanical/manual removal, biocontrols, pheromones and any other IPM methods to reduce or eliminate the pests and/or to reduce pesticide risks.

Best Management Practices (BMPs) Proposed to Reduce Pesticide Risks: If not discussed in your written plan, list planned buffers from water or sensitive habitats, wind speed restrictions, and other BMPs:

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U.S. Department of Interior Pesticide Use Proposal

Endangered Species Compliance

Federally Listed, Proposed, and Candidate Species and Designated Critical Habitat (Listed Resources): If your proposed application is located near or adjacent to any listed resources you must complete and submit th appropriate Section 7 compliance documentation as part of this PUP. For species listed by the U.S. Fish and Wildlife Service, you may complete the attached Intra-Service Section 7 form in consultation with and with assistance from the appropriate Endangered Species staff. For species listed by the National Marine Fisheries Service (NOAA Fisheries), you must contact the appropriate office and complete Section 7 consultation with them. If a determination of no effect is made, then Section 7 consultation is complete. The obligations under Section 7 must be reconsidered if: (1) new information reveals impacts of this identified action that may affect listed species or critical habitat in a manner not previously considered, (2) this action is subsequently modified in a manner that was not considered in this review, or (3) a new species is listed or critical habitat is determined that may be affected by the identified action.

Is the appropriate Section 7 documentation completed and attached? yes no N/A, no federally listed, proposed, and/or candidate species and/or critical habitats are near or adjacent to the treatment site

State-Listed Species Present:

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U.S. Department of Interior Pesticide Use Proposal

R1- 2005 0 0 PUP Reviewer(s) Signature Page

Reviewed By: Name Signature Date

Manager

Regional IPM Coordinator/Designee:

Other Reviewer if Applicable

PUP Approval/Disapproval PUP Approved As Is PUP Approved with required modifications PUP Disapproved PUP Reviewed by Region, Forwarded to WO for review

Natl. Pest Management Coordinator

PUP Approval/Disapproval PUP Approved As Is PUP Approved with required modifications PUP Disapproved

Required Modifications (please attach additional sheets if necessary):

PUP template, effective Sept. 9, 2004 R1RO ver 1.0

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APPENDIX E. USFWS Special Use Permit – Example Application

APPENDIX F. Connecticut Mosquito-Borne Virus Surveillance and Response Plans

Connecticut West Nile Virus Surveillance and Response Plan, 2012 1

State of Connecticut

West Nile Virus Surveillance and Response Plan, 2012

Introduction The West Nile Virus (WNV) Surveillance and Response Plan originally was developed in 2000 by the Mosquito Management Program (MMP), an interagency state working group led by the Department of Energy and Environmental Protection (DEEP). The Department of Public Health (DPH), the Connecticut Agricultural Experiment Station (CAES), the Department of Agriculture (DoAg), the University of Connecticut Department of Pathobiology and Veterinary Science (UConn), and the Connecticut Association of Directors of Health participated in the planning process. The Plan is used as a guide for the state’s mosquito-borne disease prevention activities.

Mosquito Management Program In 1997, Public Act 97-289, “An Act Concerning Mosquito Control and Aerial Application of Pesticides,” (CGS Sec 22a-45b) created the MMP to monitor mosquito breeding populations for the prevalence of infectious agents that can cause disease in humans and to determine when measures to abate a threat are necessary. The original focus of the program was to monitor the threat of Eastern equine encephalitis (EEE) virus. The Act authorizes the necessary measures to abate any pest-borne threat, including prevention and remedial measures, and allows for the application of broad spectrum chemical pesticides to address an imminent peril to the public health, safety, or welfare posed by pests, including mosquitoes that carry the EEE virus. The Mosquito Management Program is based on an integrated pest management (IPM) approach, which includes a combination of surveillance, education, source reduction, larval and adult mosquito control and personal protection measures.

Surveillance Activities Public health surveillance is the ongoing and systematic collection, analysis, and interpretation of health data in the process of describing and monitoring a health event. This information is used for planning, implementing, and evaluating public health interventions and programs. Surveillance activities are at the core of the Plan and currently include surveillance for EEE as well as WNV in mosquitoes, domestic animals and poultry, and humans.

Mosquito Surveillance Surveillance for WNV in mosquitoes is integral to the public health response to WNV in Connecticut. The CAES maintains a network of 91 fixed mosquito-trapping stations located in 72 municipalities throughout the state providing information that includes mosquito species composition and abundance in the community, seasonal and spatial distribution of mosquito vectors, and WNV infection rates in mosquitoes. One-third of the sites are located in southern Fairfield and New Haven counties where the highest levels of WNV activity in mosquitoes and humans have been detected in previous years (see Figs. 1a and b).

Traps are set and attended by CAES staff every 10 days at each site on a regular rotation from June through October. Two trap types are used at all trapping stations – a CO2-baited CDC Light Trap, designed to trap host-seeking adult female mosquitoes (all species), and a Gravid Mosquito Trap with hay

Connecticut West Nile Virus Surveillance and Response Plan, 2012 2

infusion, designed to trap previously blood-fed adult female mosquitoes (principally Culex and container- breeding Ochlerotatus species). Mosquitoes are transported alive to the laboratory each morning where they are identified to species. Mosquitoes are grouped (pooled) according to species, collecting site, and date and frozen at –80oC. A maximum of 50 female mosquitoes are included in each pool. Aliquots of each mosquito pool are inoculated into Vero cell cultures for detection of WNV and other mosquito-borne arboviruses of public health importance. Virus isolates from mosquito pools are tested for WNV, Eastern equine encephalitis (EEE), Jamestown Canyon (JC), Cache Valley (CV), Trivittatus (TVT), Highlands J (HJ), LaCrosse (LAC), and Potosi (POT) viruses. Isolated viruses are identified by Real Time (TaqMan) PCR or standard RT-PCR using virus-specific primers, plaque reduction neutralization (PRNT) and/or an enzyme-linked immunosorbent assay (ELISA) with specific reference antibodies. All of the virus isolation work is conducted in a certified Bio-Safety Level 3 laboratory at the CAES. Weekly test results are reported to the CDC electronically via ArboNet and to the DPH for dissemination to other state agencies, local health departments, the media, and neighboring states.

Domestic Animal Surveillance The DoAg investigates potential cases involving domestic animals, poultry and pet birds with suspicious neurologic disease reported to the State Veterinarian and/or presented for necropsy and testing to the Connecticut Veterinary Medical Diagnostic Laboratory at UConn. Horses are emphasized since they are most frequently affected. Horses presenting with the following clinical signs during the mosquito season raise suspicion: apprehension, head shaking, inability to stand, depression, flaccid paralysis of lower lip, single or multiple limb paralysis, listlessness, loss of coordination, weakness of hind limbs, or acute death. This surveillance (approximately 50,000 horses in the state) provides another means to detect the

Connecticut West Nile Virus Surveillance and Response Plan, 2012 3

presence of WNV and assess the risk of WNV infection to the human population, especially in more rural areas where mosquito trapping is not conducted. A WNV vaccine for horses is now available.

Human Surveillance The surveillance for disease in humans caused by WNV is coordinated by the DPH. Testing of serum and cerebrospinal fluid specimens for WNV antibodies and antibodies to other arboviruses (e.g. EEE, California encephalitis group, St. Louis encephalitis, Jamestown Canyon) is available at the DPH Laboratory and has been offered at no charge. Emphasis is on patients who require hospitalization for neurologic illness. Testing is available year round but is of particular importance for Connecticut residents who have not traveled during June through October indicating locally acquired infection. Physicians wishing to test persons suspected of having WNV infection on the basis of mild illness, such as fever or headache, and recent mosquito bites are encouraged to submit specimens to hospital or commercial laboratories since they are unlikely due to WNV infection and not necessary for prognostication. Reporting of positive test results from laboratories to DPH is required (see Fig. 2).

Should spraying of pesticides be conducted to reduce adult mosquito populations in response to WNV or EEE virus, the DPH also conducts surveillance for possible health effects of pesticide exposure. Physicians are encouraged to report to the DPH Environmental and Occupational Health Assessment Program possible pesticide-related health effects. The DPH compiles and summarizes this information and reports significant findings to the local health departments and other agencies as appropriate. This system is based on National Institute for Occupational Safety and Health classification of acute pesticide- related illness. The DPH assists local health departments monitor calls from the general public regarding health complaints and reports unusual clustering of complaints in terms of location or time to the DEEP Division of Pesticides for investigation of possible misapplication of pesticide.

Wild Bird Surveillance West Nile virus has been detected in dead birds of over 300 species. Infected mosquitoes carrying virus particles in their salivary glands infect susceptible bird species. Bird species capable of sustaining a sufficiently high level of virus circulating in the bloodstream for several days then serve as a source of infection for additional mosquitoes.

Although most birds infected with WNV do not develop serious illness some species, particularly crows and jays, can develop fatal infections. During the first several years of WNV in Connecticut reports of dead crows served as a useful sentinel for the presence of WNV and to describe seasonal variation. From 2000 to 2003, 92% of the human WNV infections acquired in Connecticut were preceded by a dead crow sighting in their town and 87% by a bird with laboratory confirmed WNV infection. However during 2004 and 2005, the numbers of dead crow reports and submissions for testing decreased sharply resulting in reduced utility of this system for WNV monitoring purposes.

Since 2006, mosquito surveillance has been more reliable than avian surveillance in describing the level of statewide WNV activity. Dead birds are no longer being tested for WNV. Available resources are currently devoted to maintaining the statewide mosquito trapping and testing program conducted June through October. Dead birds can be placed in a double plastic bag and placed out with the trash or brought to a municipal landfill. They can also be disposed of on-site by burying. As for all dead animals, avoid handling with birds with bare hands.

Connecticut West Nile Virus Surveillance and Response Plan, 2012 4

Mosquito Management Activities Pre-emptive mosquito control is the most effective way to prevent transmission of WNV and other mosquito-borne viruses. The most effective and economical way to control mosquitoes is by larval source reduction through local abatement programs that monitor mosquito populations and initiate control before disease transmission occurs. In addition, larval control allows for the use of target-specific agents in definable areas, which is an environmental benefit over other methods. Depending on the time of year, these programs also can be used in an emergency response for mosquito control if disease is detected in humans or domestic animals.

To prevent standing water, federal, state and local governments need to maintain existing drainage structures on their properties such as sumps, recharge basins, sewage or wastewater treatment facilities, street catch basins, upland streams, ponds, and pools. The DEEP Wetlands Habitat and Mosquito Management Program directly conducts mosquito control activities on state-owned property in coastal marsh areas and on contiguous land. The DEEP also works with municipal officials statewide to identify mosquito-breeding habitat (e.g. tidal and inland wetlands, catch basins) and develop appropriate control strategies based on Integrated Pest Management strategies to eliminate larval mosquito breeding sites.

Municipalities are responsible for coordination of mosquito control activities on municipal and private lands in their jurisdictions, working with state agencies on behalf of residents, and enforcement of abatement requirements of mosquito breeding areas if necessary. Mosquito breeding on residential and commercial properties can be reduced significantly by reducing the amount of standing water available for mosquito breeding. Regulations relevant to mosquito control and the powers of local directors of health are addressed in the Public Health Code.

To further reduce the risk of mosquito-borne virus infections, individuals are urged to take personal protective measures to avoid mosquito bites when outdoors and mosquitoes are biting through the use of repellents and proper clothing (e.g., light-colored, loose-fitting pants and shirts, head nets). Homeowners are advised to assure that window and door screens are in good repair.

Insecticides Larvicides can be used to control mosquitoes in the aquatic stage before they become biting adults. This type of control using insecticides generally is the most effective at controlling mosquitoes and has the least effect on non-target species and the environment. Ideally, use of larvicides is started early in the mosquito season repeated as necessary. The use of larvicides may require a permit from the DEEP, and the product must be registered for use in Connecticut. Depending upon the type of product used, or for commercial applications, the applicator must be licensed by the DEEP Pesticide Division to apply mosquito pesticides.

Adulticides can be used to kill adult mosquitoes when a quick reduction of mosquitoes is needed. Currently available adulticides may be applied by hand-held, backpack or truck-mounted Ultra Low Volume (ULV) foggers, or by fixed-wing or rotary aircraft. These materials have advantages and disadvantages that will influence which material is most appropriate for a given situation, and all must be applied according to regulations and label directions. Weather and logistical conditions are important factors influencing the ability to effectively control adult mosquito populations and include the following: Ground-level adulticiding is done when mosquitoes are most active (between dusk and dawn). Aerial application is done between dusk and dawn, under favorable weather conditions and at the discretion of the DEEP and its aerial contractor. Wind speed is less than 10-12 mph.

Connecticut West Nile Virus Surveillance and Response Plan, 2012 5

Wind direction and temperature inversions favor drift onto the target area. Air temperature is above 50 degrees F. Adulticide application is not made during rainfall. When making a ground-level application, the distribution and network of roads and access areas in the treatment zone are considered, as this affects the level of coverage.

Communication and Public Awareness Activities Public education about mosquito-borne diseases, particularly modes of transmitting and means of preventing or reducing risk for exposure, is a critical component of a prevention and control program. Communication and public awareness activities are designed to provide pertinent information during the mosquito season to state agencies, municipal officials, health care providers, the public, and the media including: Disease prevention recommendations including personal protective measures and homeowner source reduction. The use of larvicides, adulticides and other control methods. The virus, clinical manifestations, and its diagnosis. Mosquito Management Program information. Recommendations in response to the identification of WNV or other mosquito-borne viruses in Connecticut.

Outreach to Municipal Officials and the Public The DEEP makes available brochures, flyers, and fact sheets on WNV infection, larvicides, pesticides, personal protective measures for people, and mosquito control methods targeted at homeowners. This information and surveillance results are available in electronic format on the state Mosquito Management Program website (www.ct.gov/mosquito). The website includes links to the DPH, CAES, and DoAg. Surveillance findings are disseminated by press release to media statewide as needed. Each agency has designated staff to respond to media inquiries for up to date information. The DPH includes a WNV update as needed at its semi-annual meetings with directors of health. The DPH Infectious Diseases Section newsletter, the Connecticut Epidemiologist, is available to hospitals, laboratories, local health departments, and physicians statewide; the newsletter periodically includes summaries of prior seasons. Conference calls with local health directors occur as needed and are organized by the DPH Local Health Administration, and include members of the State Mosquito Management Team. Conference calls with state experts are a forum to discuss current state and national information and actions.

When WNV Activity is Identified Local health directors in the towns where WNV is identified in mosquitoes, domestic animals, or people are notified directly by the DPH. Notification is done by phone when the first infected mosquitoes, a human or domestic animal is identified in a town. Information available on human cases of WNV infection is posted as they become available on the DPH and web site. Upon identification of the first human case of WNV, the MMP will issue a statewide press release announcing the finding. In specific cases, where the human identification does not indicate an increase in a threat to public health, the MMP may decide not to issue a statewide press release. Examples of such cases include, but are not limited to, identification of WNV in a Connecticut resident when the infection is believed to have occurred out of state or identification of WNV in a Connecticut resident late in the season when mosquitoes are not as active and WNV is not believed to be a significant threat.

Connecticut West Nile Virus Surveillance and Response Plan, 2012 6

In coordination with officials in the towns affected, the MMP will issue a statewide press release when needed to warn Connecticut residents that WNV activity is increasing in intensity or geographic distribution and an elevated risk of human infections may occur. Throughout the season the DPH, on behalf of the MMP, will issue press releases as needed to announce findings of regional or statewide importance. The MMP conducts discussions with municipal officials in the towns affected and provides guidance for of public health actions if necessary. These may include dissemination of public information and mosquito control measures. If findings suggest a possible role for spraying to kill adult mosquitoes to mitigate a heightened risk of sustained transmission to people, the DEEP organizes and coordinates a conference call with the appropriate state and local officials. The purpose of this call is to develop a plan based on all available surveillance information and community sentiment. The state assists municipalities with key support information needed to respond to common questions from the general public. In the event adulticide use is recommended the state works with the municipality to: Notify residents of the targeted area at least 24 hours before application. Place posters and signs at key town locations as needed. Address local resident’s questions and concerns. Coordinate application with local police. Monitor health complaints.

Interagency Communication During the mosquito season state mosquito management team members are in contact regularly and multiple times each week by telephone and e-mail. Conference calls with the team occur as needed and are coordinated by the DEEP. The DEEP and DPH work together to disseminate information regarding WNV among the Mosquito Management Program agencies including surveillance results as they are available. Press releases are drafted by the DPH and distributed for review to DEEP and CAES, and, in the event of animal cases, to the DoAg.

Public Health Action Levels If WNV is confirmed in Connecticut, the DPH, in consultation with other state and local agencies, evaluate the potential threat to human health. Following evaluation of the data obtained from public health surveillance activities and depending on the nature of the threat, either the Commissioner of the DPH or the Commissioner of the DEEP, after consulting with the Commissioner of DPH, will recommend implementation of control measures.

Recommendations reflect a graded response that is in proportion to the threat of WNV infections in people. Numerous factors contribute to the level of increased risk making each situation unique. The goal is to prevent a sustained outbreak of human infections. Sporadic cases are likely to occur each season that WNV is circulating in mosquitoes since the principal mosquito species responsible for transmission is found in high numbers in residential areas.

Factors The following factors are important considerations in formulating an appropriate response to the identification of WNV: Mosquito populations and relative species abundance. Proportion of mosquitoes infected and number of pools previously identified.

Connecticut West Nile Virus Surveillance and Response Plan, 2012 7

Local surveillance data in previous season. Time of the season. Weather conditions. Geographic extent. Nature and proximity of potential mosquito habitat. Proximity and nature of human residential areas. Number and location of infected horses. Number and residence of human patients with WNV related illness. Community concern and acceptance of mosquito control activities. Extent of previous larval mosquito control activities. Likely effectiveness of local application of larval or adult insecticides.

Activities The following activities may be part of the response: Evaluation of the findings by the entire Mosquito Management Program team. Consultation with local directors of health and other municipal officials. Advice to community groups regarding outdoor activities. Dissemination of information on prevention and control methods locally or statewide. Emphasize the importance of Culex mosquito breeding site reduction on residential properties. Urge adoption of personal protective measures among high risk residents in affected areas. Expansion of mosquito trapping and human surveillance locally and beyond town lines. Identification of locations in the affected area where larviciding would be effective. Disseminate information on adulticide applications. Assessment of the need, practicality, frequency, extent and method necessary to control mosquitoes. Application of adulticide by the state with approval and at the request of municipal officials for assistance in the towns affected.

Public Health Emergency The Commissioner of DPH may proclaim a Public Health Emergency, pursuant to CGS Section 22a-66l, when WNV is confirmed in a town or contiguous towns in Connecticut. The following additional actions would be taken if a Public Health Emergency is proclaimed: In accordance with the provisions of CGS Section 28-9, the Governor evaluates the need for declaring a civil preparedness emergency. The application of adulticides by the state under these circumstances does not require the approval of the municipal officials in the towns affected. After consultation with the Commissioner of DPH, the Commissioner of DEEP has the responsibility and authority to act unilaterally if the application of chemical pesticides from the air or ground is necessary to control mosquito vectors of human disease pursuant to CGS section 22a-54(e). Concurrent with this determination, officials from the Mosquito Management Program will meet with local officials in the affected communities to inform them of the situation and to discuss the logistics of spraying.

Important State Phone Numbers and Websites Mosquito Management Program Website http://www.ct.gov/mosquito Department of Energy and Environmental Protection http://www.ct.gov/deep Communications Division (860) 424-4100

Connecticut West Nile Virus Surveillance and Response Plan, 2012 8

- State mosquito control policy and programs, media inquiries Wetlands Habitat and Mosquito Management Program (860) 642-7630 - Technical questions regarding mosquitoes, mosquito control measures Pesticide Management Program (860) 424-3369 - Technical questions regarding safe pesticide use and chemical make-up. Also, persons who wish to be specifically notified prior to a pesticide application or those who are chemically sensitive to pesticides should contact the Pesticide Pre-Notification Registry at this number Department of Public Health http://www.ct.gov/dph

Epidemiology and Emerging Infections Program (860) 509-7994 - WNV infections in people, laboratory testing of human specimens Environmental and Occupational Assessment Program (860) 509-7740 - Effects of pesticides on people Virology Laboratory (860) 509-8553 - Technical questions regarding testing of human specimens from physicians, hospitals, laboratories Connecticut Agricultural Experiment Station http://www.ct.gov/caes Main Number (203) 974-8510 - Technical questions from local health departments regarding mosquito trapping and testing University of Connecticut http://www.patho.uconn.edu Department of Pathobiology and Veterinary Science (860) 486-4000 - Testing and necropsy of animals Department of Agriculture http://www.ct.gov/doag Office of the State Veterinarian (860) 713-2505 - WNV infections in domestic animals, including livestock, poultry, and pet

Connecticut Mosquito Management Program – June, 2012

Connecticut West Nile Virus Surveillance and Response Plan, 2012 9

Connecticut West Nile Virus Surveillance and Response Plan, 2012 10

1 WNV Epidemic Curve Connecticut 1999 16 8 - 2011 0

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Figure 2. West Nile virus epidemic curve showing the cumulative number of virus-infected mosquitoes collected in surveillance traps and subsequent human cases in Connecticut from 1999 to 2008.

Contingency Plan for Eastern Equine Encephalitis (EEE)

State of Connecticut Department of Environmental Protection Connecticut Agricultural Experiment Station Department of Public Health

Routine statewide surveillance of mosquito populations for Eastern Equine Encephalitis virus activity, in conjunction with a contingency plan to control mosquito populations when necessary, will enable the state to address potential health risks in areas of concern. Consistent routine testing over a period of years will provide data upon which to revise and refine the state’s mosquito management efforts.

The state’s mosquito monitoring and management effort is a collaboration involving the Department of Environmental Protection (DEP), the Department of Public Health (DPH), and The Connecticut Agricultural Experiment Station (CAES). The program will be coordinated by the Department of Environmental Protection. DEP is responsible for the systematic identification and monitoring of mosquito breeding sites, the provision of technical assistance to municipalities and private property owners regarding mosquito control, and the collection and communication of information and data. Long term mosquito breeding site management will continue through DEP’s wetland restoration program.

The Connecticut Agricultural Experiment Station will trap, identify and submit mosquitoes for the EEE virus testing. Trapping will be conducted in areas known or suspected to support mosquito populations, which have historically tested positive for EEE, are capable of supporting such populations, or are proximate to locations where EEE-related horse deaths have occurred.

The Department of Public Health will review all mosquito test data and consult with the DEP and CAES regarding the epidemiological significance of such results. Based upon its evaluation of the potential human health risks, and in accordance with this contingency plan, DPH will advise as to appropriate personal, municipal, and state actions to reduce such risks.

Staff of DEP, DPH and CAES will form an Eastern Equine Encephalitis (EEE) Working Group to evaluate the state’s management program and protocols. This group will report to and advise the Commissioner of DEP regarding implementation of the contingency plan.

The contingency plan identifies a progression of state responses based upon Connecticut mosquito testing results, test results reported from neighboring states and reports of disease in animals in Connecticut and neighboring states. Recommended actions are limited to those that are warranted by the specific source and the extent of the potential threat to human health.

Background

Eastern Equine Encephalitis (EEE) is a rare but serious disease caused by a virus that is spread by certain kinds of mosquitoes. In Connecticut, outbreaks of EEE have occurred sporadically among horses and domestic pheasants since 1938, but no human cases have ever been confirmed. The lack of verified human cases of EEE in Connecticut is not entirely understood, since human cases have repeatedly been documented in neighboring Massachusetts and Rhode Island.

EEE is spread by mosquitoes; the transfer of the virus to a mammal can only be effected by a ‘cross bite’ scenario. That is, the mosquito must first bite a bird carrier and then a mammal. Most sites where EEE has been identified have been in or near fresh-water swamps or swamp-forest border locations that support a wide variety of wild bird life and numerous woodland mosquito species. Salt marsh mosquitoes, while they breed in huge numbers, are not generally found near the forested swamp environments where bird reservoirs of EEE are concentrated. Therefore, while the numbers of salt marsh mosquitoes and their nuisance effect is large, the risk of EEE transmission to humans is low.

At present, EEE does not appear to be a major health risk to the general public of Connecticut. An increased risk of transmission of EEE to people depends on multiple factors: introduction of EEE into swamps where there are large numbers of bird feeding mosquitoes, build up of large numbers of infected birds, isolation of EEE from multiple species of mosquitoes, isolations during the early part of the season (mid-summer) and proximity of infected mosquitoes to residential areas.

Monitoring and Public Information Program

Findings:

Basic program with no virus isolations from mosquitoes and no human, horse or commercial exotic bird deaths reported.

Actions:

• Trapping at 37* locations throughout the state will be conducted from June through October by CAES; weekly results will be reported to the media and made available to the public through an information telephone line established by the DEP. • Information regarding mosquito surveillance and control will be provided to municipal officials and local health departments by DEP. • The EEE Working Group will establish communication with Massachusetts and Rhode Island regarding their EEE monitoring programs and obtain updated information on confirmed EEE cases and public health advisories issued in those areas. • The DEP will conduct mosquito larval surveillance around the trap sites to determine the mosquito bridge vectors’ habitat areas.

* An additional 36 traps were added to test for West Nile virus and EEE in the year 2000, bringing the total of traps to 73 statewide.

Phase I: Public Health Notification

Findings:

EEE virus isolations from Culiseta melanura or other bird feeding mosquitoes (Culiseta morsitans and Culex and Culex spp.) and with human biting mosquitoes present; and/or

Confirmation of EEE virus in mosquitoes in areas of Massachusetts or Rhode Island near Connecticut borders.

Actions:

The DEP will issue a precautionary warning to appropriate local officials and health agencies, the Department of Agriculture, veterinarians (through the Connecticut Veterinary Medicine Association (CVMA), and the media for people in affected regions to avoid mosquito bites because of the potential

for increased EEE activity in Connecticut. The mosquito testing Information Line will be updated to include recommended personal protective measures.

Recommended personal protective measures to reduce mosquito bites include the following:

• Minimize outdoor activities at dawn and dusk. If you must be outdoors wear long-sleeved shirts and long pants. Use mosquito repellent that contains DEET* and follow the directions on the label. outdoor activities at dawn and dusk. If you must be outdoors wear long-sleeved shirts and long pants. Use mosquito repellent that contains DEET* and follow the directions on the label. • Cover up the arms and legs of children playing outdoors near swampy areas. When outdoors, cover babies’ playpens or carriages with mosquito netting. • Fix any holes in screens and make sure they are tightly attached to all doors and windows. • Don’t let stagnant water collect around your home, for example, in ditches, clogged gutters, old tires, wheelbarrows and wading pools. • Don’t camp overnight near freshwater swamps. When camping outdoors in tents in other areas, make sure that your tent is equipped with mosquito netting.

CAES will intensify trapping and testing in the region of occurrence. and testing in the region of occurrence.

* See Fact Sheet on DEET.

Phase II: Public Health Alert

Findings:

EEE virus isolations from human biting mosquitoes, and/or

Confirmed case of EEE involving a human, horse or commercial exotic bird.

Actions:

In addition to Phase I actions, the DEP will issue a heightened public health warning to local officials, health agencies and residents advising personal precautions in the region(s) of concern. DEP will prepare and distribute signs for posting in public places containing recommended personal precautions. In addition to the precautions recommended in Phase I, the public will be advised:

• to avoid outdoor activities from one hour before to one hour after dawn and dusk, and • not to camp out.

The Information Line will be updated to include additional measures and as necessary will include recommendations regarding the cancellation of regional outdoor activities.

The DPH will notify acute care hospitals.

The DEP will notify the Department of Agriculture and state veterinarians through the CVMA.

The EEE Working Group will evaluate the possible use of Ultra Low Volume (ULV) fogging application to knock down mosquito vectors within the region of the trap site and/or the application of larvicide to known mosquito bridge vectors larval breeding areas. The isolation of the virus from bird

feeding mosquitoes alone does not pose a significant health threat to the public that would warrant pesticide spraying. Any recommendation to use pesticide will be determined by consideration of the weather conditions, the number of virus isolations, mosquito species with EEE virus, mosquito population estimates, and breeding cycles.

• Pesticide applications to be made by State of Connecticut staff and/or by certified commercial pest control operators. • Affected communities, including municipal officials and the general public, shall be notified in advance of any pesticide application.

The EEE Working Group, if necessary, will determine the most appropriate method of adulticide application: aircraft or truck mounted adulticiding. Truck mounted application is limited by access to the intended area while aerial application can affect large areas and numbers of adult mosquitoes but is less discriminating, exposes more people and wildlife, and is more expensive and less easily repeated. Any decision to apply pesticides from trucks or the air would be made only after evaluation of the multiple factors which contribute to risk of transmission of EEE to people and after discussion with officials from the potentially affected community.

STATE OF CONNECTICUT Zika Virus Surveillance and Response Plan, 2016 Revised 04/22/2016

Introduction

The Zika Virus Surveillance and Response Plan is based on the West Nile Virus Surveillance and Response Plan updated in 2012 by the Mosquito Management Program (MMP), an interagency state working group led by the Department of Energy and Environmental Protection (DEEP). The purpose of this Plan is to provide a guide for the state’s Zika virus prevention activities. The Plan will be modified and updated as additional information and federal guidance regarding this newly emerging threat becomes available. Some provisions of the Plan depend on availability of additional state or federal funds.

Zika virus, first discovered in Uganda in 1947, was limited to Africa and Asia infrequently causing human illnesses. In May 2015, the World Health Organization reported the first local transmission of Zika virus in the Western Hemisphere. As of April 8, 2016, local transmission has been identified in at least 34 countries or territories in the Americas, including Puerto Rico, with further spread to other countries in the region likely but precisely to what extent is currently not known.

In the Western Hemisphere Zika virus is transmitted primarily by Aedes aegypti, the mosquito that also spreads yellow fever, dengue and chikungunya viruses. Aedes aegypti are aggressive daytime biters, can also bite at night and can be found in buildings. They become infected with Zika virus when they bite a person already infected with the virus and can then transmit the virus when they bite another person. While Ae. aegypti is not present in Connecticut, a related species, Aedes albopictus, has been identified in the southwestern area of the state and it is also considered capable of transmission of the virus. The degree to which it will contribute to transmission in Connecticut is not known. Of recent concern is also the possibility of spread from a woman to her baby during pregnancy and between sexual partners.

Historically and in the majority of recent case-patients, Zika virus causes asymptomatic infections or relatively mild illnesses that are rarely fatal. However, an association with Guillain-Barre syndrome has been suggested and, during the current outbreak, an increase in birth defects among infants born to women infected during pregnancy is associated with the virus. The full spectrum of clinical manifestations is not known.

Mosquito Management Program

In 1997, Public Act 97-289, “An Act Concerning Mosquito Control and Aerial Application of Pesticides,” (CGS Sec 22a-45b) created the MMP to monitor mosquito breeding populations for the prevalence of infectious agents that can cause disease in humans and to determine when measures to abate a threat are necessary. The original focus of the program was to monitor the threat of Eastern equine encephalitis (EEE) virus. The Act authorizes the necessary measures to abate any pest-borne threat, including prevention and remedial measures, and allows for the application of broad spectrum chemical pesticides to address an imminent peril to the public health, safety, or welfare posed by pests. The Mosquito Management Program is based on an integrated pest management (IPM) approach, which includes a combination of surveillance, education, source reduction, larval and adult mosquito control, and personal protection measures.

Based on the multiple modes of potential transmission, severe health consequences to neonates, the role of laboratory testing for diagnosis and medical management, and heightened public concern, Governor Dannel P. Malloy designated the Department of Public Health (DPH) as the lead agency for the State’s response to Zika virus. The DPH will also be responsible for conducting surveillance for human cases of Zika virus associated illnesses and coordinating dissemination of information. The Department of Energy and Environmental Protection (DEEP) will provide technical advice regarding mosquito control to municipalities and The Connecticut Agricultural Experiment Station will conduct mosquito surveillance and provide entomological expertise.

Communications

As the designated lead agency for Connecticut’s response to Zika virus, the DPH will coordinate communications. This will include disseminating information on: 1) the risk of acquiring infection in Connecticut during the mosquito season, 2) results of human and mosquito surveillance, 2) precautions that women and men should take to avoid mosquito bites when travelling to an affected area, 3) national guidelines for prevention of fetal infections, 4) availability of laboratory testing, and 4) mosquito control activities for control of Ae. albopictus.

Principal methods to inform the public and health care providers will include: • DPH Zika virus web page including links to the Mosquito Management Program web site and to other state and national resources • Press releases and, potentially, press conferences for important announcements • Radio public service announcements in English, Spanish and Portuguese • The DPH Health Alert Network (HAN) which provides electronic real-time communications to health care providers, local health departments, first responders and others • The Connecticut Epidemiologist newsletter • Existing DPH Programs such as WIC, Healthy Start, Immunization Action Coordinators • Advisories at Bradley International Airport for travelers to and from affected areas

Surveillance Activities

Public health surveillance is the ongoing and systematic collection, analysis, and interpretation of health data in the process of describing and monitoring a health event. This information is used for planning, implementing, and evaluating public health interventions and programs. Surveillance activities are at the core of the Plan and currently include surveillance for Zika virus in humans and mosquitoes.

Human Surveillance

The surveillance for disease in humans caused by Zika virus is coordinated by the DPH. Zika virus was recently declared a nationally notifiable disease. Zika virus disease was added to lists of Reportable Diseases, Emergency Illnesses and Health Conditions and Reportable Laboratory Findings in Connecticut effective Monday, February 15, 2016. As of 4/8/2016, there are no commercially available diagnostic tests; testing for Connecticut residents is conducted by the DPH Public Health Laboratory (PHL) or at the Centers for Disease Control and Prevention (CDC). Specimens are submitted to the DPH PHL for testing or shipment to the CDC. Required patient demographic, clinical and travel history is collected by the DPH Epidemiology and Emerging Infections Program (EEIP) using a standardized questionnaire. Staff of the EEIP is available 24/7/365 to answer questions and facilitate testing of appropriate specimens. The DPH Laboratory reports test results directly to the requesting health care provider.

During the first week after onset of symptoms, Zika virus disease can often be diagnosed by performing reverse transcriptase-polymerase chain reaction (RT-PCR) on serum. Virus-specific IgM and neutralizing antibodies typically develop toward the end of the first week of illness; cross-reaction with related flaviviruses (e.g., dengue and yellow fever viruses) is common and may be difficult to discern. Plaque- reduction neutralization testing can be performed to measure virus-specific neutralizing antibodies and often discriminate between cross-reacting antibodies in primary flavivirus infections.

Beginning March 1, 2016, the DPH PHL was able to begin offering RT-PCR testing for Zika virus genetic material. On April 11, 2016 identification of IgM antibodies was added to the list of tests available. These two types of tests account for most of the testing that is needed. Some specimens will continue to be sent to CDC for testing when additional test methodologies (e.g. PRNT) are needed for confirmation of results or testing of tissue specimens (e.g. immunohistochemistry).

Recommendations for testing are evolving. The current emphasis is on testing of serum specimens from pregnant women with a history of travel to areas where Zika virus is circulating in the prior 2-12 weeks for the presence of IgM antibodies. Testing of serum specimens by RT-PCR is also available regardless of pregnancy status for symptomatic patients if collected during the first week of illness. As more is learned about transmission and the spectrum of illnesses caused by Zika viruses, testing protocols will be modified.

A Zika virus module was developed in the Connecticut Electronic Disease Surveillance System (CT EDSS) database to monitor provider reports of Zika virus associated illness, travel exposures of pregnant women and laboratory test results. DPH Laboratory test results are imported electronically from the DPH Laboratory information management system into CT EDSS. Local health directors have web based access to jurisdiction-specific information based on the patient’s official town of residence.

To identify children with potential Zika virus associated birth defects, laboratory and provider reporting will be supplemented by regular review of the Connecticut Birth Defects Registry. The registry collects information on birth defects through various sources of data, including reporting from birth hospitals across the state, vital records, and hospital discharge data. Testing will be encouraged for children born with microcephaly, poorly developed brain structures or ocular defects not attributed to other causes, and their mothers

Should spraying of pesticides be conducted to reduce adult mosquito populations in response to Zika, WN or EEE viruses, the DPH also conducts surveillance for possible health effects of pesticide exposure. Physicians are encouraged to report to the DPH Environmental and Occupational Health Assessment Program possible pesticide-related health effects. The DPH compiles and summarizes this information and reports significant findings to the local health departments and other agencies as appropriate. This system is based on National Institute for Occupational Safety and Health classification of acute pesticide- related illness. The DPH assists local health departments monitor calls from the general public regarding health complaints and reports unusual clustering of complaints in terms of location or time to the DEEP Division of Pesticides for investigation of possible misapplication of pesticide.

Mosquito and Virus Surveillance

Surveillance for Zika virus in mosquitoes is integral to the public health response in Connecticut by identifying the areas where local transmission, though unlikely, may potentially take place. The Connecticut Agricultural Experiment Station (CAES) maintains a network of 91 fixed mosquito-trapping stations located in 72 municipalities throughout the state providing information that includes mosquito species composition and abundance in the community, seasonal and spatial distribution of mosquito vectors, and prevalence of virus infected mosquitoes. One-third of the sites are located in southern Fairfield and New Haven counties where the highest levels of West Nile virus (WNV) activity in

mosquitoes and humans have been detected in previous years. Transmission of WNV is primarily by Culex pipiens mosquitoes which, like Ae. albopictus, lays its eggs in small containers of water often found near homes.

Traps are set and attended by CAES staff every 10 days at each site on a regular rotation from June through October. At least two trap types are used at the trapping sites designed to collect host-seeking adult female mosquitoes (all species) and previously blood-fed adult female mosquitoes (principally Culex and container-breeding Aedes species). Mosquitoes are transported alive to the laboratory each morning where they are identified to species. Mosquitoes are grouped (pooled) according to species, collecting site, and date and then frozen. Aliquots of each mosquito pool are inoculated into Vero cell cultures for detection of mosquito-borne arboviruses of public health importance. Isolated viruses are identified by Real Time (TaqMan) PCR or standard RT-PCR using virus-specific primers. All of the virus isolation work is conducted in a certified Bio-Safety Level 3 laboratory at the CAES. Weekly test results are reported to the CDC electronically via ArboNet and to the DPH for dissemination to other state agencies, local health departments, the media, and neighboring states.

Based on trapping during 2006-2015, distribution of Ae. albopictus is confined to the southwestern portion of Connecticut but it has been expanding up the coast and increasing in abundance. Currently areas of potential concern are south of I-95 in the 10 coastal towns from Greenwich to West Haven. Collection numbers are low compared to other species, but may be due to under sampling because the current traps are not efficient for luring and trapping Ae. albopictus. Expansion of the trapping program to specifically target Ae. albopictus would include addition of BG-Sentinel traps baited with human scent

lure and placed in locations in Connecticut where Ae. albopictus has been collected in previous years or in peripheral sites with the appropriate habitat for this species. For the 2016 season, BG-Sentinel traps will be deployed once Ae. albopictus is identified at existing trap sites.

The CAES has expanded diagnostic testing capabilities to include detection of Zika virus in field-collected mosquitoes. The cell culture assay currently used is ideal for screening for a diversity of arboviruses and was found to be very sensitive for Zika virus. An RT-PCR assay was recently optimized to test for Zika virus at CAES laboratories. The need and feasibility of molecular diagnostic tools to improve sensitivity and timeliness for detection of Zika virus directly from mosquito pools will be explored in subsequent seasons.

Prevention Activities

Environmental Control

The primary mode of transmission is by mosquito bites. Therefore, pre-emptive mosquito control is the most effective way to prevent transmission of Zika virus and other mosquito-borne viruses. The most effective and economical way to control mosquitoes is by larval source reduction through local abatement programs that monitor mosquito populations and initiate control before disease transmission occurs. With similar preferences for breeding habitat, efforts to reduce Cx. pipiens

populations for control of WNV will also reduce populations of Ae. albopictus. In Connecticut, municipalities are responsible for coordination of mosquito control activities on municipal and private lands in their jurisdictions, working with state agencies on behalf of residents, and enforcement of abatement requirements of mosquito breeding areas if necessary. Technical advice regarding source reduction is available for municipalities from the DEEP Wetlands Habitat and Mosquito Management Program. Assistance from the DEEP is focused on training municipal personnel to conduct mosquito control activities in their jurisdictions. Direct participation from the DEEP is limited and depends on allocation of state funds for field staff.

Larvicides can be used to control mosquitoes in the aquatic stage before they become biting adults. This type of control using insecticides generally is the most effective at controlling mosquitoes and has the least effect on non-target species and the environment. Ideally, use of larvicides is started early in the mosquito season and repeated as necessary. The use of larvicides may require a permit from the DEEP, and the product must be registered for use in Connecticut. Depending upon the type of product used, or for commercial applications, the applicator must be licensed by the DEEP Pesticide Division to apply mosquito pesticides. Recommended larvicide use is as per Strategies for the Application of Larvicides to Control Mosquitoes in Response to West Nile Virus in Connecticut (updated and approved by DEEP, DPH, CAES, DoAG in January, 2014). The following options are available.

 Products containing the biological agents Bacillus sphaericus (Bs) or Bacillus thuringiensis var. israelensis (Bti). Both agents come in a granular, wetable powder, slow release briquette or water-soluble packet formulations. Also available are dual-action formulations of Bs and Bti. The bacterial strains in Bs are more specific to Culex larvae than Bti. Bs and Bti are bacterial agents and must be ingested by the filter-feeding mosquito larvae and as such, these products will not affect the non-filter feeding mosquito pupae or adults. The use of Bti or Bs on municipal or individual homeowner property does not require any special licensing by the CT DEEP. • S-methoprene (trade name Altosid). Methoprene is an insect growth regulator and comes in a variety of liquid, granular, pellet and briquette formulations. If using Altosid for catch basins a pellet, 30-day or 150-day briquette formulation is recommended. Methoprene will not affect pupae or adults. Connecticut regulations specify that the use of methoprene requires that the applicator be licensed and a permit be obtained from the DEEP prior to application. Also, PA 13-197 prohibits certain uses of methoprene in the coastal zone (http://www.cga.ct.gov/2013/ACT/PA/2013PA-00197-R00HB-06441- PA.htm). • The biological agent Saccharopolyspora spinosa or Spinosad (trade name Natular®). Spinosad comes in a variety of formulations and works on all mosquito species. Natular will not affect mosquito pupae or adults. Although it is a bacterial agent, because of its mode of action, Connecticut regulations specify that the use of spinosad requires that the applicator be licensed and a permit be obtained from the DEEP prior to application. • The Larvasonic Acoustic Larvacide Device emits sound waves to kill mosquito larvae (www.newmountain.com). The Larvasonic works on all species of mosquitoes. Mosquito larvae must be present for the Larvasonic to be effective and as such, requires more intensive larval surveillance. Since this device works by emitting sound waves, it is not considered a pesticide and therefore is exempt from pesticide regulations.

be applied according to regulations and label directions. Weather and logistical conditions are important factors influencing the ability to effectively control adult mosquito as well as the preferred habitat and feeding habits of the target mosquito species.

Preventing Mosquito Bites

There is no available vaccine to prevent Zika virus infection and no specific treatment for Zika virus related illnesses. Prevention depends on avoiding mosquito bites. When travel cannot be avoided to countries where Zika virus or other viruses spread by mosquitoes are found people should take the following steps:  Weather permitting wear long-sleeved shirts and long pants; loose fitting light colors work best  Stay in places with air conditioning or that use window and door screens to keep mosquitoes outside  Sleep under a mosquito bed net if you are overseas or outside and are not able to protect yourself from mosquito bites  Use Environmental Protection Agency (EPA)-registered insect repellents. When used as directed, EPA-registered insect repellents are proven safe and effective, even for pregnant and breast-feeding women. o DEET o Picaridin (also known as KBR 3023) o Oil of lemon eucalyptus (OLE) or PMD (Products containing OLE include Repel and Off! Botanicals) o IR3535 o Always follow the product label instructions o Reapply insect repellent as directed o Do not spray repellent on the skin under clothing o If you are also using sunscreen, apply sunscreen before applying insect repellent  If you have a baby or child: o Do not use insect repellent on babies younger than 2 months of age o Dress your child in clothing that covers arms and legs, or o Cover crib, stroller, and baby carrier with mosquito netting o Do not apply insect repellent onto a child’s hands, eyes, mouth, and cut or irritated skin o Adults: Spray insect repellent onto your hands and then apply to a child’s face.  Treat clothing and gear with permethrin or purchase permethrin-treated items o Treated clothing remains protective after multiple washings - see product information to learn how long the protection will last o If treating items yourself, follow the product instructions carefully o Do not use permethrin products directly on skin  If you have Zika, protect others from getting sick, avoid mosquito bites during the first week of illness

Prevention of Transmission of Zika Virus in Women of Reproductive Age

(National recommendations as of 4/11/2016) Pregnant women should avoid travel to areas where Zika virus is circulating locally among mosquitoes for the duration of the pregnancy whenever possible. When travel cannot be avoided women should:  Practice prevention measures for preventing mosquito bites

 Be tested for the presence of virus by RT-PCR during the first 7 days - if she develops illness • Be tested for the presence of IgM antibodies 2-12 weeks after the last potential exposure to help guide prenatal care – if she does not develop illness

Recommendations for men and their pregnant partners In addition to bites of infected mosquitoes, sexual transmission from men to sexual partners is also possible and is of particular concern for women who are pregnant or intend to become pregnant. At this time, tests of serum or semen of men for the purpose of assessing risk for sexual transmission is not recommended due to lack of understanding of what such test results mean. As more is learned about the incidence and duration of seminal shedding from infected men recommendations will be updated. Based on what is currently known about transmission men who travel to an area of active Zika virus transmission should:  Abstain from sex (i.e., vaginal intercourse, anal intercourse, or fellatio) for the duration of the pregnancy or  Consistently and correctly use condoms  Have pregnant partner tested if either partner develops symptoms

Recommendations for men and their non-pregnant sex partners Since most infections are asymptomatic or cause mild illness among adults, mosquito-borne Zika virus infections may go unrecognized. For persons who travel to a Zika affected area and want to attempt conception:  Women who become ill should wait at least 8 weeks after symptom onset  Women who do not become ill should wait at least 8 weeks from last date of exposure  Men who are diagnosed with Zika virus disease should wait at least 6 months  Men who do not become ill should wait at least 8 weeks from last date of exposure

Phased Risk-Based Response

Prevention activities in Connecticut adapted from CDC national recommendations are appropriate to the potential levels of risk for transmission of Zika virus in Connecticut or for travelers to Zika virus affected areas. They are based on human and environmental surveillance systems and include:

Before mosquito season Response Actions • Designation of DPH as lead agency to coordinate the State’s response  Identification of agency response personnel at DPH, DEEP and CAES  Develop a state response plan specific for Zika virus based on West Nile and Eastern equine encephalitis plans and national recommendations  DPH Commissioner is briefed weekly on planning progress Communication • Include DPH updates for local health departments on the regularly scheduled monthly statewide conference calls – on 2/18/2016 the entire hour long call was devoted to Zika with participation of the DEEP and CAES  On 5/20/2016 the CAES will host a half-day symposium on vector-borne infectious diseases for local health directors and selected staff  Conduct public mosquito education campaigns focusing on reducing or eliminating larval habitats for other mammal-biting container breeding mosquitoes including Ae. albopictus (DEEP, DPH, municipalities) • DEEP Mosquito Management Program will update and maintain the state’s web site: www.ct.gov/mosquito with information on mosquito-borne illness in humans, mosquito

surveillance and control options, and the Zika virus Response Plan.  The DPH will maintain a Zika virus page on its web site  The DPH will develop public service announcements for radio on mosquito source reduction, prevention of fetal infections, prevention of sexual and mosquito transmission to pregnant women or women who plan to conceive  Train DPH Epidemiology and Emerging Infections Program staff to respond to telephone inquiries from the public and health care providers, and collect information on Zika virus associated illnesses including fetal infections Surveillance  Zika virus added to the lists of reportable diseases and laboratory findings on 2/15/2016  Develop surveillance system for human cases including a standardized questionnaire module in CT EDSS  Implement electronic reporting of Zika virus test results from DPH Laboratory to the EEIP  Use the Newborn Screening and Birth Defects Registry systems to identify newborns with potential Zika virus associated birth defects born to pregnant women who were not previously tested  Designate DPH contacts for the US Pregnancy Registry Laboratory  Develop capacity to conduct testing of human specimens for Zika virus  DPH Laboratory offers RT-PCR testing starting 3/1/2016  DPH Laboratory offers IgM ELISA testing starting 4/11/2016 Vector Control  Recommend surveys to determine abundance, distribution, and type of containers that may serve as mosquito breeding sites, especially in towns where Ae. albopictus has been previously identified (municipalities with technical advice/training provided by DEEP)  Cover, dump, modify or treat large water-holding containers with long-lasting larvicides (municipalities, property owners)  DEEP Pesticide Management Program will prioritize registration of products and the issuance of permits needed for the commercial application of pesticides and insecticides.  DEEP Solid Waste Program will assist with outreach and education efforts and will prioritize coordination with local officials to address blight and illegal disposal of materials such as tires. Will pursue enforcement actions involving large-scale tire disposal areas. Pregnant women outreach  Distribute guidance for testing potentially exposed pregnant women, prevention of mosquito and sexual transmission and monitoring of developing fetuses  Develop radio PSA  Use the HAN to communicate directly with obstetricians/gynecologists and other licensed health care providers • “Piggy-back” on existing DPH Programs that distribute information to pregnant women and their health care providers such as WIC, Healthy Start, Immunization Action Coordinators

Beginning of mosquito season Activities conducted before mosquito season are continued and, in addition, the following are added if not already in place. Response Actions

 As for WN and EEE viruses regular telephone and email communications are conducted between designated staff of the DPH, DEEP and CAES Communication  Continue public education campaigns focusing on reducing or eliminating larval habitats for Ae. albopictus (DPH, DEEP) • Notify the local director of health of human cases or infected mosquitoes in the health department’s jurisdiction (DPH)  Announce identification of imported cases (DPH)  Distribute information about Ae. albopictus and personal protection measures (DPH, LHDs) to avoid mosquito bites among travelers returning from Zika affected areas, including posters at Bradley International Airport, urging: o Insect repellents o Window and door screens to prevent mosquitoes from entering the house o Air conditioning  Elimination of larval habitats at least within 100-200 yards/meters around the residence of imported cases (municipality) • Results of mosquito trapping and testing are posted on the CAES web site with links from the MMP, DPH and DoAg – these include weekly results at each trap site and maps with positive findings Surveillance  Mosquito trapping is conducted June through October (CAES) to: o estimate abundance of mosquito species o determine distribution o develop detailed vector distribution maps o evaluate the efficacy of source reduction and larvicide treatment  If Ae. albopictus is identified at established trapping sites, BG-Sentinel traps will be used to further monitor populations Vector Control  Based on integrated pest management  Continued emphasis on larval source reduction and use of larvicides in large containers that cannot be emptied (municipalities)

Single or several local mosquito-acquired cases Activities conducted with the start of mosquito season are continued and, in addition, the following are added if not already in place Response Actions  Determine the need for assistance from a CDC field team to provide technical, communication, vector control, or logistical support Communication  Notify Red Cross  Distribute updates utilizing multiple methods including press releases and the DPH HAN Vector Control  Consideration will be given to adult mosquito control in consultation with Director of Health and Chief Elected Official in affected town o Treat the outdoors within 100–200 yards/meters around a case’s home o Initiate/maintain adult sampling to estimate adult mosquito abundance and evaluate effectiveness of insecticide treatments o Use of adulticides will consider the following factors:

 Considerations for use of adulticides o Abundance of Ae. albopictus o Number of infected pools o Local surveillance data in previous season o Time of the season o Weather conditions o Geographic extent o Proximity and nature of human residential areas o Number and residence of human patients with Zika virus related illness o Community concern and acceptance of mosquito control activities o Extent of previous larval mosquito control activities o Likely effectiveness of local application of insecticides Pregnant women outreach  Enhanced targeted warnings to pregnant women regarding prevention of mosquito bites

Outbreak - clusters cases This is considered very unlikely during the 2016 season in Connecticut. CDC recommendations include:  Divide the outbreak area into operational management areas where control measures can be effectively applied to all buildings and public areas within a few days; repeat as needed to reduce mosquito density  Conduct door-to-door inspections and mosquito control in an area-wide fashion  Identify and treat, modify, or remove mosquito-producing containers  Organize area/community clean-up campaigns targeting disposable containers (source reduction), including large junk objects that accumulate water (broken washing machines, refrigerators, toilets) in buildings, public areas, etc. • Combine outdoor spatial or residual spraying with source reduction and larviciding (including residual spraying of container surfaces and adjacent mosquito resting areas). Don’t forget to treat storm drains, roof gutters and other often overlooked cryptic water sources

Important State Phone Numbers and Websites State Mosquito Management Program Website http://www.ct.gov/mosquito

Department of Public Health http://www.ct.gov/dph

Office of Communications (860) 509-7270 -Media inquiries Epidemiology and Emerging Infections Program (860) 509-7994 - Zika virus infections in people, laboratory testing of human specimens Environmental and Occupational Assessment Program (860) 509-7740 - Effects of pesticides on people Public Health Laboratory (860) 920-6500 - Technical questions regarding testing and shipping of human specimens from physicians, hospitals, laboratories

Department of Energy and Environmental Protection http://www.ct.gov/deep Communications Division (860) 424-4100 - State mosquito control policy and programs

Wetlands Habitat and Mosquito Management Program (860) 642-7630 - Technical questions regarding mosquitoes, mosquito control measures

Pesticide Management Program (860) 424-3369 - Technical questions regarding safe pesticide use and chemical make-up. Also, persons who wish to be specifically notified prior to a pesticide application or those who are chemically sensitive to pesticides should contact the Pesticide Pre-Notification Registry at this number

Connecticut Agricultural Experiment Station http://www.ct.gov/caes Center for Vector Biology & Zoonotic Diseases (203) 974-8510 - Technical questions regarding mosquito trapping and testing

F-46

APPENDIX G. Connecticut Trapping and Arbovirus Testing Program

Center for Vector Biology & Zoonotic Diseases

The Center for Vector Biology & Zoonotic Diseases at The Connecticut Agricultural Experiment Station (CAES) brings together the research, surveillance and diagnostic activities of our scientific and technical staff working on arthropods of public health and veterinary importance and the infectious disease organisms they transmit in Connecticut and the northeastern United States. The mission of the Center is to advance knowledge on the ecology and epidemiology of vector-borne disease organisms and to develop novel methods and more effective strategies for their surveillance and control.

The Center is responsible for conducting the statewide Mosquito and Arbovirus Surveillance Program for eastern equine encephalitis (EEE) and West Nile (WNV) viruses and testing of ticks for the Lyme disease agent. Scientists at the Center are also engaged in full-time laboratory and field research on the biology and control of mosquitoes, ticks and bedbugs, and are investigating the epidemiology and ecology of a variety of mosquito- and tick-associated diseases that occur throughout the region including: eastern equine encephalitis, human babesiosis, ehrlichiosis, granulocytic anaplasmosis, Lyme disease, West Nile virus, Powassan virus and a related deer tick virus.

The Center maintains several microbiology, pathology, immunology, electron microscopy and molecular biology laboratories located at the main campus in New Haven and a Biosafety Level 3 containment facility where a world-wide reference collection of about 475 arboviruses are housed. The laboratory is one of the few in the northeast that is certified by the Centers for Disease Control and Prevention and the US Department of Agriculture to work with "select agents". Select agents are bio-agents which have been declared by the U.S. Department of Health and Human Services or by the U.S. Department of Agriculture to have the "potential to pose a severe threat to public health and safety". The Tick Identification, Testing and Information Laboratory and Insect Information Office are also located in New Haven, while insectary facilities for maintaining insect, tick, and vertebrate animal colonies are located at the Station's 75-acre research farm, Lockwood Farm in Hamden, CT. A 28- acre field station and laboratory for conducting additional studies is located at the Griswold Research Center in Griswold/Voluntown, CT. Core funding for the Center is provided from the State of Connecticut and federal Hatch

2 funds administered by the U.S. Department of Agriculture. Research and surveillance activities on mosquitoes and mosquito-borne diseases are additionally supported in part by an "Epidemiology and Laboratory Capacity for Infectious Diseases (ELC)" grant from the Centers for Disease Control and Prevention (CDC) administered through the Connecticut Department of Public Health (DPH). The Center has nine lead scientists, one support scientist and seven technicians, currently divided into three Research Groups investigating (1) Mosquitoes, (2) Ticks and (3) Bedbugs. A description of the Mosquito Research and Surveillance Program is described below.

Mosquito Research and Surveillance Group Scientists: • Dr. Theodore Andreadis, Director - Medical Entomology • Dr. Philip Armstrong, Scientist - Virology • Dr. Douglas Brackney, Assistant Scientist - Virology • Dr. Goudarz Molaei, Associate Scientist - Insect Physiology and Molecular Biology • Dr. Charles Vossbrinck, Associate Scientist - Evolution, Ecology and Molecular Biology • Dr. John Anderson, Distinguished Scientist Emeritus - Medical Entomology Support Staff • John Shepard, Assistant Scientist I – Mosquito laboratory, New Haven • Angela Bransfield, Technician II and Responsible Official – BSL3 laboratory, New Haven • Michael Thomas, Technician II – Mosquito laboratory, New Haven • Michael Misencik, Technician I – BSL3 laboratory, New Haven • Michael Vasil, Technician II – Mosquito and animal colony laboratory, Lockwood farms • Heidi Stuber, Technician I – Mosquito and animal colony laboratory, Lockwood farms

Mosquito Research and Surveillance Programs Program History

In 1997, Public Act 97-289, An Act Concerning Mosquito Control and Aerial Application of Pesticides, (CGS Sec 22a-45b) created a Mosquito Management Program to monitor mosquito breeding populations for the prevalence of infectious agents that can cause disease in humans and to determine when measures to abate a threat are necessary. The original focus of the program was to monitor the threat of EEE virus in southeastern CT and state funding was given to the Experiment Station to create a state-wide mosquito trapping and testing program. The original program, the first of its kind in the State, was established by Dr. Theodore Andreadis and included 36 trapping locations where approximately 50,000 mosquitoes were trapped and tested on an annual basis from June through October. State funding was used to hire one Technician and five seasonal staff to assist in the collection and identification of mosquitoes. Virus isolation work was subcontracted out to Yale University, as no Biosafety Level 3 (BSL3) laboratory for conducting such work existed at the Experiment Station at that time. The following year, a small laboratory at the Experiment Station which had been used for Rocky Mountain spotted fever and Lyme disease studies, was converted to a BSL3 virus isolation laboratory and certified by the CDC and DPH, as facilities at Yale were closing. Dr. John Anderson, then Experiment Station Director, assumed responsibilities for operation of the virus laboratory and a second laboratory Technician was hired to assist in this work.

The establishment of the mosquito trapping and testing program proved to be most timely, as it allowed Experiment Station staff to respond immediately to the introduction of WNV into the

3 US in 1999. This led Station scientists to obtain the first isolations of WNV from mosquitoes in North America, an achievement that was published in the prestigious journal Science (Dec. 17, 1999).

The unprecedented establishment and expansion of WNV in the US necessitated an expansion of the mosquito trapping and testing program to include more densely populated urban and suburban communities in the State where the peridomestic mosquitoes that carried the virus were most abundant and where WNV activity proved to be a greater threat to human health. With grants from the US Department of Agriculture (Congressional award) and CDC (ELC funds) the program was expanded threefold to include trapping at 91 locations throughout the State. These sites were mostly located in southeastern and central CT. Two new Assistant Scientists, 2 Postdoctoral Scientists, 5 Technicians and 8 additional seasonal staff were hired with this funding to sustain the expanded surveillance program and support new research initiatives on WNV. The USDA funding was additionally used in 2003 to equip a new state-of-the-art BSL3 laboratory in the Johnson-Horsefall building with expanded capacity to handle increased numbers of mosquitoes and new mosquito-borne viruses. Permanent cuts in CDC funding ($230,000 to $110,000 annual) which directly supported the surveillance program occurred in 2009, and all USDA funding ($750,000 annual) which supported both surveillance and research programs was eliminated in 2010 with the abolishment of many congressional programs. This resulted in the termination of 2 Postdoctoral Scientists and 2 Technicians and a scaling back of specific research programs. State funding was increased in 2011 from $230,000 to $460,000 to make up for the loss in federal funding and to specifically maintain the surveillance program at its current level thus avoiding elimination of trapping locations in the State.

Current Surveillance Program Description

West Nile and EEE viruses constitute ongoing threats to human health in Connecticut causing severe illness and death. Surveillance for EEE and WNV in mosquitoes is integral to the public health response to these mosquito-borne diseases and the Mosquito Trapping and Testing Program conducted by the Experiment Station provides an effective early warning system that directs targeted intervention strategies and prevents disease in humans and domestic animals. The objectives of the surveillance program are to provide:

• Early evidence of local virus activity, including new viruses that may emerge • Information on the abundance, distribution, identity and infection rates of mosquito vectors • Information that is used to assess the threat of WNV virus and EEE to the public and guide the implementation of mosquito control measures

The entire state-wide program, including collection and identification of mosquitoes and isolation and identification of viruses is undertaken by staff at the Experiment Station under the direction of Dr. Philip Armstrong who also supervises virus isolation work in the BSL3 laboratory. Support staff include: 4 laboratory technicians and eleven seasonal stuff that assist in mosquito collection in the field and identification in the laboratory, virus isolation and identification in the BSL3 laboratory and electronic data management and reporting to the DPH and the National CDC Arbovirus Surveillance System (ArboNet).

4 The CAES maintains a network of 91 permanent mosquito-trapping stations in 72 municipalities throughout the state. One- third of the sites are located in densely populated areas in southern Fairfield and New Haven counties where the highest levels of WNV activity in mosquitoes and humans are detected each year. Another third are located in southeastern CT where EEE acticity is most frequently encountered. Annual training of seasonal support staff begins in May and formal mosquito trapping and testing continues through October. Mosquito trapping is conducted daily and at each site every 10 days on a regular rotation until virus is detected. Thereafter, trapping frequency is increased to 2-3 times per week in an effort to obtain more information on the abundance of specific mosquito vectors and virus activity.

Two trap types are used at all trapping stations - a CO2-baited CDC Light Trap, designed to trap host-seeking adult female mosquitoes (all species), and a Gravid Mosquito Trap with hay infusion, designed to trap previously blood-fed adult female mosquitoes (principally Cu/ex that transmit WNV).

CO2-baited CDC Gravid Mosquito Trap Light Trap

Mosquitoes are transported alive to the laboratory each morning where they were identified to species. Mosquitoes are grouped (pooled) according to species, collecting site, and date and frozen at -80°C. A maximum of 50 female mosquitoes are included in each pool. Aliquots of each mosquito pool are inoculated into Vero cell cultures (African Green Monkey cells) and screened daily for detection of WNV, EEE and other mosquito-borne arboviruses of public health importance. Virus isolates from mosquito pools are tested for WNV, EEE, Jamestown Canyon (JC), Cache Valley (CV), Trivittatus (TVT), Highlands J (HJ), Lacrosse (LAC), and Potosi (POT) viruses. Isolated viruses are identified by Real Time (TaqMan) polymerase chain reaction (PCR) or standard RT-PCR using virus-specific primers, plaque reduction neutralization (PRNT) and/or an enzyme-linked immunosorbent assay (ELISA) with specific reference antibodies. All of the virus isolation work is conducted in the certified Bio-Safety Level 3 laboratory at the CABS. Complete processing of mosquitoes is usually completed within 7days. Weekly test results are immediately reported to the CDC electronically via ArboNet and to the DPH for dissemination to other state agencies, local health departments, the media, and neighboring states.

Since 1996, nearly than 2.8 million mosquitoes have been trapped and tested and specific geographic localities in the State and time of the season associated with increased risk of human exposure to EEE and WNV have been identified. A total of 1,402 isolations of WNV have been made from 21 different species of mosquitoes , and a total of 341 isolations of EEE have been made from 18 species of mosquitoes. There have been 110 human cases of WNV in the state with 3 fatalities and one human case (fatal) of EEE. The principal foci of WNV activity in Connecticut have been identified as densely populated residential communities in coastal Fairfield and New Haven Counties. The principal foci for EEE activity are in more rural locales located in the southeastern comer of the state. We have observed a correlation both temporally and spatially between the isolation of WNV and EEE from field-collected mosquitoes and the elevated risk of human infection that typically extends from late July through September in Connecticut. Seasonal Activity of West Nile Virus

200 20 ..------Human- - - Cases------r -+-WNV (+) Mosquitoes

160 16

(1) 0" ' '5 120 12 CT u"' "' C :£ E"' 8 :i i 80

40 4

Human Cases of West Nile Virus in Connecticut 2000 - 2013

2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 2013

Mosquito Collection and Arbovirus Testing Summary 1996-2013

No. Mosquitoes No. Pools No. Arboviruses Isolated Year Trapped Tested CV EEE HJ JC LAC PV TVT WNV 36 1996 6,440 620 - 19 - - - - -

1997 45,556 3,268 - 2 22 7 - - - -

1998 66,383 3,981 22 8 23 5 - - 2 - 1 9 1999 45,391 3,524 - - - - 1 2 1 10 11 24 20 2000 137,199 9,085 - 3 14 7 2001 124,414 10,135 3 14 31 6 - - 30 16 4 2002 119,950 10,643 - - - - - 73 15,447 20 72 2003 255,334 72 87 56 - 73 16 12,521 37 12 1 2004 156,409 - - - - 43 9,843 2005 111,731 1 - - 22 1 - 4 34 12,661 4 3 5 23 3 2006 197,793 - 7 219 11,233 5 2 42 69 2007 157,476 - - - - 15,108 13 13 20 163 9 2008 211,657 - - 191 16,895 1 122 61 43 35 2009 291,641 - - 29

2010 115,377 10,580 6 4 1 22 - - - 220 19,132 64 2 9 166 2011 333,334 43 4 21 . 2012 189,379 14,058 14 234 - 9 3 . 6 -

2013 192,172 13,601 - 58 9 15 - 3 IO 90 2,759,025 192,568 163 399 234 400 1 270 102 1,493 TOTAL

CV= Cache Valley, EEE = Eastern Equine Encephalitis, HJ = Highlands J, JC= Jamestown Canyon, LAC= Lacrosse, PV = Potosi, TVT = Trivittatus, WNV = West Nile Virus

Current Research Projects on Mosquitoes and Mosquito-Borne Diseases

The Connecticut Agricultural Experiment Station has a distinguished history of research on mosquitoes, ticks and associated disease-causing agents. Our scientists made the first isolation of WNV in North America and were instrumental in assisting the CDC Division of Vector-Borne Infectious Diseases during the initial outbreak (GAO/HEHS-00-180, West Nile Virus Outbreak: Lessons for Public Health Preparedness, September 2000) Since then, we have remained on the forefront of efforts to investigate and monitor mosquito-borne viruses and tick associated diseases throughout the Northeast through research and surveillance activities conducted at our Center for Vector Biology & Zoonotic Diseases.

Since its introduction in 1999, WNV virus has sickened nearly 30,000 people across the US and Canada resulting in over 1,500 fatalities. It is estimated that in the US alone, more than 1.6 million people have been infected and new evidence indicates that the virus can persist for years in convalescing patients. In 2012 we experienced the largest outbreak ever recorded in the State with 234 WNV positive mosquito pools recovered from 51 different locations in 44 towns in the slate and a record 21 human cases. Meanwhile, we have witnessed a resurgence of EEE virus activity throughout the northeastern US, including regions where it had not been previously detected, resulting in 26 human cases and 9 fatalities. Our surveillance activities have been integral to the public health response to these viruses and have provided an early warning system that has directed targeted intervention strategies and prevented transmission of mosquito-borne diseases to humans. More than two and a half million mosquitoes have been trapped and tested and specific geographic localities in the state and time of the season associated with increased risk of human exposure have been identified. Work in our Select Agent Laboratory, where a worldwide reference collection of arboviruses is currently held, has resulted in the isolation and molecular characterization of seven mosquito-borne viruses that cause human disease, including two not previously recognized in the state, LaCrosse and Potosi. This work highlights our capacity to detect and respond to the potential threat to the U.S. from mosquito transmitted diseases such as dengue, chickungunya and Rift Valley fever viruses which affect millions of people globally. Through our research initiatives, we have further:

• Elucidated the natural history and epidemiology of WNV in the northeastern US including the role of various mosquitoes and birds • Evaluated the competence of mosquitoes to transmit and serve as over-wintering hosts for the virus • Examined the feeding and biting behavior of the primary mosquito vectors of WNVand other mosquito-borne viruses • Developed more sensitive and rapid molecular diagnostic techniques to identify viruses • Documented the introduction and establishment of two invasive mosquitoes from Asia • Developed and tested novel mosquito trapping methods to enhance the early detection of mosquito-borne viruses • Evaluated the efficacy of new and established biological agents to control mosquitoes.

The findings from these investigations have resulted in the publication of over 95 scientific articles in 26 different peer-reviewed journals since 1999.

A comprehensive surveillance program complemented with science-based mosquito control and a well-developed and timely outreach network continue to be the most effective ways to educate and protect the public. The recent establishment of invasive mosquitoes (Aedes albopictus and Aedesjaponicus) from Asia in the continental U.S. including Connecticut, and the potential for the introduction of exotic mosquito-borne viruses are cause for concern due to increased global trade and travel. The detection of emerging threats from abroad in our Select Agent Virus Laboratory is further strengthened by our strategic location in close proximity to the nation's busiest ports and travel hubs, New York and Boston. We are the only regional agency in the northeastern US with the facilities, biological reference collections, technical scientific expertise, trained personnel and infrastructure to conduct these investigations from one central location. Our research findings have national significance and we are collaborating with researchers and agencies throughout the nation including: California, Delaware, Florida, Georgia, Illinois, Kentucky, Massachusetts, Michigan, New Hampshire, New Jersey, New Mexico, New York, North Carolina, and North Dakota.

Current Research Projects include:

West Nile Virus • Continuing to monitor the ecology and epidemiology of WNV in the northeastern US • Tracking changes in mosquito populations and virus activity patterns that may be related to global climate change • Examining the role of various mosquitoes in supporting enzootic virus transmission among avian hosts and epidemic "bridge" transmission to humans • Investigating virus titer variation in field-collected mosquitoes to evaluate their capacity to acquire, replicate and transmit the virus • Defining genetic mutations in WNV that may impact virulence, host-range, and infectivity through whole genome sequencing

Eastern Equine Encephalitis • Investigating the re-emergence and expansion of eastern equine encephalitis virus in the northeastern US • Examining the role of Culiseta melanura and wild avian hosts in supporting local enzootic virus transmission (amplification) in several recognized freshwater foci in Connecticut (Chester, Killingworth, Madison, North Stonington) and epidemic "bridge" transmission to mammals • Examining regional differences in the epidemiology of EEE virus activity in the northeast. Have cooperative projects with Vermont, Massachusetts and New York • Studying the fine scale-scale population structure of EEE virus along the eastern seaboard through molecular phylogenetic analyses to infer virus introduction, spread, overwintering and extinction in the region

• Examining the ecological and climatic factors associated with the re-emergence of eastern equine encephalitis virus in the northeast. Cooperative project with Dr. Heidi Brown, University of Arizona • Examining the genetic structure of Culiseta melanura populations throughout the east coast using novel DNA-based nucleotide polymorphism markers (microsatellites) to determine if population differences exist that may be associated with vector competence • Identified a unique mosquito-specific flavivirus within Culiseta melanura populations in Connecticut. Studies are underway to characterize this virus through molecular and ultrastructural analyses and assess its distribution among geographically isolated populations

Human Cases of EEE in the Northeastern US 1965- 2013

11 Exotic Invasive Mosquitoes Ochlerotatus (Aedes) japonicus • 1998 - First detected in State, likely introduced in mid-l 990's • Shown to be established throughout State • Evaluating the competitive impact of the species on native mosquito species in natural and artificial container habitats • Evaluating the biological control potential of a novel recently described microsporidian parasite from Japan, Takaokaspora nipponicus • Evaluating its role as a local vector of WNV and other arboviruses

Aedes albopictus • 2006 - first adults detected in Stratford and at a tire recycling plant in Sterling, but unable to overwinter • 2012 - Overwintering populations confirmed in Stratford for first time • Tracking its dispersal throughout coastal Fairfield and New Haven Counties. Current distribution extends from Greenwich to West Haven.

PUBLICATIONS ON MOSQUITOES AND MOSQUITO-BORNE DISEASES 1999-2014

1999 Anderson, J. F., Andreadis, T. G., Vossbrinck, C.R., Tirrell, S, Wakem, E. M., French' R. A., Garmendia, A. E. and Van Kruiningen, H.J. 1999. Isolation of West Nile Virus from mosquitoes, crows, and a Cooper's hawk in Connecticut. Science, 286:2331-2333. Andreadis, T. G. 1999. Epizootiology Amblyospora stimuli (Microspora: Amblyosporidae) infections in field populations of a univoltine mosquito, Aedes stimulans (Diptera: Culicidae) inhabiting a temporary vernal pool J. Invertebr. Pathol. 74:198-205. Andreadis, T. G., Anderson, J. F., and Vossbrinck. 1999. Mosquito arbovirus surveillance in Connecticut, 1999: isolation and identification of West Nile virus. Proc. Northeastern Mosq. Control Assoc. 45:57-67.

2000 Garmendia, A. E. Van Kruiningen, H.J., French, R. A, Anderson, J. F., Andreadis, T. G., Kumor, A and West, A. B. 2000. Recovery and identification of West Nile virus from a hawk in winter. J Cinical Microbial. 38:31 l 0-3111.

2001 Anderson, J. F., Vossbrinck, C. F., Andreadis, T. G., Iton, A. Beckwith, W. H. and Mayo, D. R. 2001. A phylogenetic approach to following West Nile virus in Connecticut. Proc. Nat. A.cad. Sci. 98: 12885-12889. Anderson, J. F., Vossbrinck, C. F., Andreadis, T. G., lton, A. Beckwith, W. H. and Mayo, D. R. 2001. Characterization of West Nile virus from five species of mosquitoes, nine species of birds, and one mammal. Ann. NY Acad Sci. 951:328-331. Andreadis, T. G., Anderson, J. F. and Vossbrinck, C. F. 2001. Mosquito surveillance for West Nile virus in Connecticut, 2000: isolation from Culex pipiens, Cu/ex restuans, Cu/ex salinarius and Cu/iseta melanura. Emerg. Infect. Dis. 7:670-674. Andreadis, T. G., Anderson, J. F., Munstermann, L. E., Wolfe, R. J. and Florin, D. A. 2001. Discovery, distribution and abundance of a newly introduced mosquito, Ochlerotatus japonicus (Diptera: Culicidae) in Connecticut, USA. J Med. Entomol. 38:774-779.

Eidson, M. and the West Nile Avian Mortality Surveillance Group [Andreadis, T., Anderson J., Vossbrinck, C.]. 2001. Crow deaths as a sentinel surveillance system for West Nile virus in the northeastern United States, 1999. Emerg. Infect. Dis. 7:615-620. Fonseca, D. M., Campbell, S., Crans, W. J., Mogi, M, Miyagi, I., Toma, T., Bullians, M., Andreadis, T. G., Berry, R. L. Pajac, B., Sardelis, M. and Wilkerson, R. C. 2001. Aedes (Finlaya) japonicus (Diptera: Culicidae) a newly recognized mosquito in the USA: first analyses of genetic variation in the US and putative source populations. J. Med. Entomol. . 38:133-146. Hadler, J., Nelson, R., McCarthy, T., Andreadis, T., Lis, M. J., French, R., Beckwith, W., Mayo, D., Archambault, G. and Cartter, M. 2001. West Nile virus surveillance in Connecticut, 2000: evidence that an intense epizootic can occur without humans being at high risk for severe disease. Emerg. Infect. Dis. 7:636-642. Marfin, A.A. and the ArboNET Cooperative Surveillance Group [Andreadis, T.]. 2001. Widespread West Nile virus activity, eastern United States, 2000. Emerg. Infect. Dis. 7:730- 735. Micieli, M. V., Garcia, J. J. and Andreadis, T. G. 2001. Epizootiological studies of Amblyospora albifasciati (Microsporidiida: Amblyosporidae) in natural populations of Aedes albifasciatus (Diptera: Culicidae) and Mesocyclops annulatus (Copepoda: Cyclopidae) in a transient floodwater habitat. J Invertebr. Pathol. 77:68-74. Wang, T., Anderson, J. F., Magnarelli, L.A., Bushmich, S., Wong, S. Koski, R. A., and Fikrig, E. 2001. West Nile virus envelope protein-role in diagnosis and immunity. Ann. NY Acad Sci. 951: 325-327.

Wang, T., Anderson, J. F., Magnarelli, L.A., Wong, S., Koski, R., and Fikrig, E.. 2001. Immunization of mice against West Nile virus with recombinant envelope protein antisera. Journ. Immunol. 167:5273-5277.

2002 Anderson, J. F. and Rahal, J. J. 2002. Efficacy oflnterferon Alpha-2B and Ribavirin against West Nile virus in-vitro. Emer. Infect. Dis. 8:107-108. Andreadis, T. G. 2002. Epizootiology of Hyalinocysta chapmani (Microsporidia: Thelohaniidae) infections in field populations of Culiseta melanura (Diptera: Culicidae) and Orthocyclops modestus (Copepoda: Cyclopidae): a three-year investigation. J. Invertebr. Pathol. 81:114-121. Andreadis, T. G. 2002. West Nile virus: an exotic emerging pathogen in the New World. VIIIth Intl. Colloq. Invertebr. Pathol. and Microbial Control. Foz do Iguassu, Brazil. pp. 58- 64. Andreadis, T. G. and Vossbrinck, C. F. 2002. Life cycle, ultrastructure and molecular phylogeny of Hyalinocysta chapmani (Microsporidia: Thelohaniidae) a parasite of Culiseta melanura (Diptera: Culicidae) and Orthocyclops modestus (Copepoda: Cyclopidae). J. Euk. Microbial. 49:350-364. Wang T., Magnarelli, L.A., Anderson, J. F., Gould, L. H., Bushmich, S. L., Wong, S. J., and Fikrig, E. 2002. A recombinant envelope protein-based enzyme-linked immunosorbent assay for West Nile virus serodiagnosis. Vector Borne Zoonotic Dis. 2:105-9.

2003 Anderson, J. F., Main, A. J., Andreadis, T. G., Wikel, S. K. and Vossbrinck, C.R. 2003. Transstadial transfer of West Nile virus by four species of ixodid ticks (Acari: Ixodidae). J. Med. Entomol. 40:528-533. Andreadis, T. G. 2003. A checklist of the mosquitoes of Connecticut with new state records. J. Am. Mosq. Control Assoc. 19:79-81. Andreadis, T. G., Becnel, J. J., and White, S. E. 2003. Infectivity and pathogenicity of a novel baculovirus, CuniNPV from Culex nigripalpus (Diptera: Culicidae) for thirteen species and four genera of mosquitoes. J. Med. Entomol. 40:512-517. Armstrong, P. M. and Rico-Hesse, R.. 2003. Efficiency uf dt:ngut: st:rutypt: 2 virus strains to infect and disseminate in Aedes aegypti. Am. J. Trop. Med. Hyg. 68:539-544. Wang T., Scully, E., Yin, Z., Kim, J. H., Wang, S., Yan, J., Mamula, M., Anderson, J. F., Craft, J., and Fikrig, E. 2003. IFN-gamma-producing gamma delta T cells help control murine West Nile virus infection. J. Immunol. 171:2524-31.

2004 Anderson, J. F., Andreadis, T. G., Main, A. J., and Kline, D. L. 2004. Prevalence of West Nile virus in tree-canopy inhabiting Culex pipiens and associated mosquitoes. J. Trop. Med Hyg. 71: 112-119.

Andreadis, T. G., Anderson, J. F., Vossbrinck, C.R. and Main, A. J. 2004. Epidemiology of West Nile virus in Connecticut, USA: a five year analysis of mosquito data 1999- 2003. Vector-Borne and Zoonotic Dis. 4:360-378.

Rahal, J. J., J. F. Anderson, C. Rosenberg, T. Reagan, and L.L. Thompson. Effect of interferon alpha-2b therapy on St. Louis virus meningoencephalitis: clinical and laboratory results of a pilot study. J. Infect. Dis. 190:1084-1087, 2004.

Spielman, A., Andreadis, T. G., Apperson, C. S., Cornel, A. J., Day, J. F., Edman, J. D., Fish, D., Harrington, L. C., Kiszewski, A. E., Lampman, R., Lanzaro, G. C., Matuschka, F. R., Munstcrmann, L. E., Nasci, R. S., Norris, D. E., Novak, R. J., Pollack, R. J., Reisen, W. K., Reiter, P., Savage, H. M., Tabachnick, W. J., and Wesson, D. M. 2004. Outbreak of West Nile virus in North America. Science. 306:1473-5. Vossbrinck, C.R., Andreadis, T. G., Vavra, J. and Becnel, J. J. 2004. Molecular phylogeny and evolution of mosquito parasitic Microsporidia (Microsporidia: Amblyosporidae). J. Euk. Microbiol. 51:88-95.

Wang, T, T. Town, L. Alexopoulou, J. F. Anderson, E. Fikrig, and R. A. Flavell. 2004. Toll like receptor 3 mediates West Nile virus entry into the brain causing lethal encephalitis. Nature Medicine 10: 1366-1373, 2004.

2005 Andreadis, T. G. 2005. Evolutionary strategies and adaptations for survival between mosquito parasitic microsporidia and their intermediate copepod hosts: a comparative examination of Amblyospora connecticus and Hyalinocysta chapmani (Microsporidia: Amblyosporidae). Folia Parasitologica 52: 23-35. Andreadis, T. G., Thomas, M. C. and Shepard, J. J. 2005. Identification guide to the mosquitoes of Connecticut. Conn. Agric. Exp. Stn. Bull. 617, 179 pp. Armstrong, P. M., Andreadis, T. G., Anderson, J. F. and Main, A. J. 2005. Isolations of Potosi virus from mosquitoes (Diptera: Culicidae) collected in Connecticut. J. Med Entomol. 42:875-881. Farajollahi, A., Crans, W. J., Nickerson, D., Bryant, P., Wolf, B., Glaser, A. and Andreadis, T. G. 2005. Detection of West Nile virus RNA from the louse fly Icosta americana (Diptera: Hippoboscidae). J. Am. Mosq. Control Assoc. 21:474-476. Turell, M. J., Dohm, D. J., Sardelis, R., O'Guinn, M. L., Andreadis, T. G. and Blow, J. A. 2005. An update on the potential of North American mosquitoes (Diptera: Culicidae) to transmit West Nile virus. J. Med Entomol. 42:57-62. Tuttle, A. D., Andreadis, T. G., Frasca, S. Jr. and Dunn, J. L. 2005. Eastern equine encephalitis in a flock of African penguins maintained at an aquarium. J. Am. Vet. Med Assoc. 226:2059- 2062.

2006 Anderson, J. F., Andreadis, T. G., Main, A. J., Ferrandino, F. J., and Vossbrinck, C.R. 2006. West Nile virus from female and male mosquitoes (Diptera: Culicidae) in subterranean, ground, and canopy habitats in Connecticut. J. Med. Entomol. 43: 1010-1019.

Anderson, J. F. and G., Main, A. J. 2006. Importance of vertical and horizontal transmission of West Nile virus by Culex pipiens in the northeastern United States. J. Infect. Dis. 194:1577-1579.

Armstrong, P. M. and Andreadis, T. G. 2006. A new genetic variant of La Crosse virus (Bunyaviridae) isolated from New England. Am. J. Trap. Med. Hyg. 75:491-496.

Diuk-Wasser, M.A., Brown, H. E., Andreadis, T. G. and Fish, D. 2006. Modeling the spatial distribution of mosquito vectors for West Nile virus in Connecticut, USA. Vector-Borne and Zoonotic Dis. 6:283-295.

Molaei, G. and Andreadis, T. G. 2006. Identification of avian- and mammalian-derived blood meals in Aedes vexans and Culiseta melanura (Diptera: Culicidae) and its implication for West Nile virus transmission in Connecticut, USA. J. Med. Entomol. 43:1088-1093.

Molaei, G., Andreadis, T. G., Armstrong, P. M., Anderson, J. F. and Vossbrinck, C.R. 2006. Host feeding patterns of Culex mosquitoes and West Nile virus transmission, northeastern United States. Emerg. lrifect. Dis. 12:468-474.

Molaei, G., Oliver, J., Andreadis, T. G., Armstrong, P. M., and Howard, J. J. 2006. Molecular identification of blood meal sources in Culiseta melanura and Culiseta morsitans from an endemic focus of eastern equine encephalitis (EEE) virus in New York, USA. Am. J. Trap. Med Hyg. 75:1140-1147.

Shepard, J. J., Andreadis, T. G. and Vossbrinck, C.R. 2006. Molecular phylogeny and evolutionary relationships among mosquitoes (Diptera: Culicidae) from the northeastern United States based on small subunit ribosomal DNA (18S rDNA) sequences. J. Med Entomol. 43:443-454.

Wang, T., Y. Gao, E. Scully, C. T. Davis, J. F. Anderson, A. Barrett, M. Ledizet, R. Koski, A. Yin, J. Craft, and E. Fikrig. γδ T cells facilitate adaptive immunity against West Nile virus infection in mice. J. Immun. 177:1825-1832, 2006.

2007

Anderson, J. F., Main, A. J. Ferrandino, F. J. and Andreadis, T. G. 2007. Nocturnal activity of mosquitoes (Diptera: Culicidae) in a West Nile focus in Connecticut. J. Med. Entomol. 44:1102-1108.

Andreadis, T. G. 2007. Microsporidian parasites of mosquitoes. In: T. G. Floore (ed.), Biorational Control of Mosquitoes, Bull. No. 7, Am. Mosq. Control Assoc. 23:3-29.

Andreadis T. G. and Armstrong P. M. 2007. A 2-yr evaluation of elevated canopy trapping for Cu/ex mosquitoes and West Nile virus in an operational surveillance program in the northeastern United States. J. Am. Mosq. Control Assoc. 23: 137-148.

Armstrong, P. M. and Andreadis, T. G. 2007. Genetic relationships of Jamestown Canyon virus strains infecting mosquitoes collected in Connecticut, USA. J. Trap. Med. Hyg. 77:1157-116 Bai, F, T. Town, D. Pradhan, J. Cox, Ashish, M. Ledizet, J. F. Anderson, R. A. Flavell, J. K. Krueger, R. A. Koski, and E. Fikrig. 2007. Antiviral peptides targeting the West Nile virus envelope protein. J Virol. 81: 2047-2055. Linthicum, K. J., Anyamba, A., Britch, S. C., Chretien, J.P., Erickson, R. L., Small, J., Tucker, C. J., Bennett, K. E., Mayer, R. T., Schmidtmann, E.T., Andreadis, T. G., Anderson, J. F., Wilson, W. C., Freier, J.E., James, A. M., Miller, R. S., Drolet, B. S., Miller, S. N., Tedrow, A., Baile y, C. L., Strickm an, D. A., Barnard, D. R., Clark, G. G., and Zou, L. 2007. A Rift Valley fever risk surveillance system for Africa using remotely sensed data: potential for use on other continents. Vet. Ital. 43:663-674.

Molaei, G., Andreadis, T. G., Armstrong, P. M., Bueno, R., Dennett, J., Bala, A., Randle, Y., Guzman, H., Da Rosa, A. T., Wuithiranyagool, T., and Tesh, R. B. 2007. Host feeding pattern of Cu/ex quinquefasciatus (Diptera: Culicidae) and its role in transmission of West Nile virus in Harris County, Texas. Am. J Trop. Med. Hyg. 77: 73- 81.

16 . 2008 Anderson, J. F., A. J. Main, K. Delroux, and E. Fikrig. 2008. Extrinsic incubation periods for horizontal and vertical transmission of West Nile virus by Culex pipiens pipiens. J Med.Entomol. 45: 445-451

Andreadis, T. G., J. F. Anderson, P. M. Armstrong, and A. J. Main. 2008. Isolations of Jamestown Canyon virus (Bunyaviridae: Orthobunyavirus) from field-collected mosquitoes (Diptera: Culicidae) in Connecticut, USA: a ten-year analysis, 1997-2006. Vector-Borne and Zoonotic Dis. 8:175-188.

Armstrong, P. M., Andreadis, T. G., Anderson, J. F. and Mores, C. N. 2008. Tracking eastern equine encephalitis virus perpetuation in northeastern U.S. by phylogenetic analysis. Am. J Trop. Med Hyg. 79:291-296.

Brown, H. E., Diuk-Wasser, M.A., Andreadis, T. G. and Fish, D. 2008. Remotely-sensed vegetation indices identify mosquito clusters of West Nile virus vectors in an urban landscape in the northeastern US. Vector-Borne Zoonotic Dis. 8:197-206.

Coleman, A. S, Yang, X., Kumar, M., Zhang, X. Promnares, K., Shroder, D., Kennedy, M. R., Anderson, J. F, Akins , D.R., and Pal, U. 2008. Borrelia burgdorferi Complement Regulator-Acquiring Surface Protein 2 Does Not Contribute to Complement Resistance or Host Infectivity. PLOS ONE (8) art.e3010

Huang, S., Molaei, G., and Andreadis, T. G. 2008. Genetic insights into the population structure of Cu/ex pipiens (Diptera: Culicidae) in the northeastern United States by using microsatellite analysis. Am. J Trop. Med Hyg. 79:518-527.

Kong, K-F., X. Wang, J. F. Anderson, E. Fikrig, R.R. Montgomery. 2008. West Nile virus attenuates activation of primary human macrophages. Viral Immunol. 21:78-82.

Magnarelli, L. A., S. L. Bushmich, J. F. Anderson , M. Ledizet, and R. A. Koski. Serum antibodies to West Nile virus in naturally exposed and vaccinated horses . J Med. Microbiol. 57:1087-1093.

Molaei, G., Andreadis, T. G., Armstrong, P. M., and Duik-Wasser, M. 2008. Host-feeding patterns of potential mosquito vectors of arboviral agents in Connecticut, USA: molecularanalysis of blood meals from twenty-three species of Aedes, Anopheles, Culex, Coquillettidia, Psorphora, and Uranotaenia. J. Med. Entomol. 45:1143-1151.

Simakova, A. V., Vossbrinck, C.R. and Andreadis, T. G. 2008. Molecular and ultrastructural characterization of Andreanna caspii n. gen., n. sp. (Microsporidia: Amblyosporidae) a parasite of Ochlerotatus caspius (Diptera: Culicidae). J. Invertebr. Pathol. 99:302-311.

Wang, S., T. Welte, M. McGargill, T. Town, J. Thompson, J. F. Anderson, R, A, Flavell, E. Fikrig, S. M. Hedrick, and T. Wang. 2008. Drak2 signaling mediates T cell trafficking and helps West Nile virus entry into the brain. J. Immunology. 181:2084-91.

Welte, T, J. Lamb, J. F. Anderson, W. K. Born, R. L. O'Brien, M. Zabel, and T. Wang. 2008. The role of two distinct Gamma/delta T cell subsets during West Nile virus infection. FEMS Immunol. Med. Microbiol. 53:275-283.

2009

Anderson, J. F., F. J. Ferrandino, S. Mcknight, J. Nolen, and J. Miller. A carbon dioxide, heat, and chemical lure trap for capturing bed bugs, Cimex lectularius. 2009. Med. Vet. Entomol. 23:99-105.

17 Andreadis, T. G. 2009. Failure of Aedes albopictus (Diptera: Culicidae) to overwinter following introduction and seasonal establishment at a tire recycling plant in the northeastern USA. J. Amer. Mosq. Control Assoc. 25: 25-31.

Andreadis, T. G. 2009. Trapping and testing program for mosquito-borne viruses to begin June 1. Connecticut Weekly Agricultural Report 89: 1-3.

Bonafe, N., J. A. Rininger, R. G. Chubet, H. G. Foellmer, S. Fader, J. F. Anderson, S. L. Bushmich, K. Anthony, M. Ledizet, E. Fikrig, R. A. Koski, and P. Kaplan. 2009. A recombinant West Nile virus envelope protein vaccine candidate produced in Spodoptera frugiperda expresSF+ cells. Vaccine. 27:213-222.

Huang, S., Hamer, G., L., Molaei, G., Walker, E., D., Goldberg, T., L., Kitron, U. D., and Andreadis, T. G. Genetic variation associated with mammalian feeding in Cu/ex pipiens from a West Nile virus epidemic region in Chicago, Illinois. Vector-Borne Zoonotic Dis. 9:637-642.

Liu, A., Lee, V., Galusha, D., Slade, M., Diuk-Wasser, M., Andreadis, T., Nelson, R., Scotch, M., and Rabinowitz, P. Risk factors for human infection with West Nile virus in Connecticut: a multi-year·analysis. Int. J. Health Geographies 8:67 doi:10.l 186/1476- 072X- 8-67.

Micieli, M. V., Garcia, J. J. Andreadis, T. G. 2009. Factors affecting horizontal transmission of the microsporidium Amblyospora albifasciati to its intermediate host, Mesocyclops annulatus. J. Invertebr. Pathol. 101:228-233.

Molaei, G., Farajollahi, A., Armstrong, P. M., Oliver, J., Howard, J. J., and Andreadis,T. G. 2009. Identification of blood meals in Anopheles quadrimaculatus and Anopheles punctipennis from eastern equine encephalitis virus foci in northeastern USA. Med. Vet. Entomol. 23:350-356.

Molaei, G., Farajollahi, A, Scott, J. J., Gaugler, R., and Andreadis, T. G. 2009. Human blood feeding by the recently introduced mosquito, Aedes japonicus japonicus (Diptera: Culicidae) and public health implications. J. Amer. Mosq. Control Assoc. 25:210-214.

Simpson, J.E., Folsom-O'Keefe, C. M., Childs, J.E., Simons, L. E., Andreadis, T. G. Diuk Wasser, M.A. 2009. Avian host-selection by Culexpipiens Say (Diptera: Culicidae) in experimental trials. PLOS One 4(11): e7861. doi:10.1371/journal.pone.0007861.

Town, T., F. Bai, T. Wang, A. T. Kaplan, F. Qian, R.R. Montgomery, J. F. Anderson, R. A. Flavell, E. Fikrig. 2009. Tlr7 mitigates lethal West Nile encephalitis by affecting inte4rleukin 23- dependent immune cell infiltration and homing. Immunity. 30:242-253.

2010

Andreadis, T. G., Armstrong, P.A., and Bajwa, W. J. 2010. Studies on hibernating populations of Cu/ex pipiens (Diptera: Culicidae) from a West Nile virus endemic focus in New York City: parity rates and isolation of West Nile virus. J. Am. Mosq. Control Assoc. 26:257-264.

Andreadis, T. G. and Wolfe, R. J. 2010. Evidence for reduction of native mosquitoes with increased expansion on the by the invasive Ochlerotatus japonicus japonicus (Diptera: Culicidae) in the northeastern United States J. Med. Entomol. 47:43-52.

Armstrong, P.A. and Andreadis, T. G. 2010. Eastern equine encephalitis virus in mosquitoes and their role as bridge vectors. Emerging Inf Dis. 16:1869-1874

Diuk-Wasser, M.A., Molaei, G., Simpson, J.E., Folsom-O'Keefe, C. M., Armstrong, P. M., and Andreadis, T. G. Avian communal roosts as amplification foci for West Nile virus in urban areas in northeastern United States. Am. J. Trop. Med Hyg. 82:337-343.

Molaei, G., Cummings, R. F., Su, T., Armstrong, P. M., Williams, G. A., Cheng, M. L., Webb, J.P., and Andreadis, T. G. 2010. Vector-host interactions governing epidemiology of West Nile virus in southern California. Am. J. Trop. Med. Hyg. 83:1269-1282

Vossbrinck, C.R., Baker, M. D., and Andreadis, T. G. 2010. Phylogenetic position of Octosporea muscaedomesticae (Microsporidia) and its relationship to Octosporea bayeri based on small subunit rDNA analysis. J. lnvertebr. Pathol. 105:366-370.

2011

Anderson, J. F., Ferrandino, F. J., Dingman, D. W., Main, A. J., Andreadis, T. G., and Becnel, J. J. 2011. Control of mosquitoes in catch basins in Connecticut with Bacillus thuringiensis israelensis, Bacillus sphearicus and spinosid. J. Am. Mosq. Control Assoc. 27:45-55.

Armstrong, P. M, Andreadis, T. G., Finan S., Shepard, J. J., Thomas, M. C., and Anderson, J. F. 2011. Detection of infectious virus from field-collected mosquitoes by Vero cell culture assay. J. Visualized Exper. 52. http://www.jove.com/index/Details.stp?ID=2889,doi: 10.3791/2889.

Armstrong, P. M., Vossbrinck, C.R., Andreadis, T. G., Anderson, J. F., Pesko, K. N., Newman, R. M., Lennon, N. J., Birren, B. W., Ebel, G.D. and Henn, M. R. 2011. Molecular Evolution of West Nile virus in a northern temperate region: Connecticut, USA 1999-2008. Virol. 417:203-210.

Huang, S., Molaei, G., and Andreadis, T. G. 2011. Reexamination of Cu/ex pipiens hybridization zone in the eastern United States by ribosomal DNA-based single nucleotide polymorphism markers. Am. J Trop. Med. Hyg. 85:434-441.

Simakova, A. V., Luk'iantsev, V. V., Vossbrinck, C.R., Andreadis, T. G. 2011. Identification of mosquito-parasitic microsporidia, Amblyospora rugosa and Trichoctosporea pygopellita (Microsporidia: Amblyosporidae), from Acanthocyclops venustus and Acanthocyclops reductus (Copepoda: Cyclopidae), based on small subunit rDNA analysis. Parazitologiia 45:140-146 (in Russian with English abstract).

19 2012

Anderson, J. F., Main, A. J., Cheng, G., Ferrandino, F. J. and Fikrig. E. 2012. Horizontal and vertical transmission of West Nile Virus genotype NY99 by Culex salinarius and genotypes NY99 and WN02 by Cu/ex tarsalis. Am. J Trop. Med. Hyg. 86:134-139.

Anderson, J. F., McKnight, S. and Ferrandino, F. J. 2012. Aedesjaponicusjaponicus and associated woodland species attracted to CDC miniature light traps baited with carbon dioxide and the Traptech mosq utio lure. J Amer. Mosq. Control Assoc. 28:184-191.

Andreadis, T. G. 2012. The contribution of Culex pipiens complex mosquitoes to transmission and persistence of West Nile virus in North America. J Amer. Mosq. Control Assoc. 28s:137-151.

Andreadis, T. G. and Armstrong, P. M. 2012. 2012: a record year for West Nile virus activity in Connecticut. CT Weekly Agric. Report 92:1-4.

Andreadis, T. G., Shepard, J. J. and Thomas, M. C. 2012. Field observations on the overwintering ecology of Culiseta melanura in the northeastern United States. J Amer. Mosq. Control Assoc. 28:286-291

Andreadis, T. G., Simakova, A. V., Vossbrinck, C. R, Shepard, J. J., Yurchenko, Y. A. 2012. Ultrastructural characterization and comparative phylogenetic analysis of new Microsporidia from Siberian mosquitoes: evidence for coevolution and host switching. J lnvertebr. Pathol. 109:59-75.

Armstrong, P. M., Prince, N. and Andreadis, T. G. 2012. Development of a multi-target TaqMan assay to detect eastern equine encephalitis virus variants in mosquitoes. Vector Borne Zoonotic Dis. 12:872-876.

Hardstone, M. C. and Andreadis, T. G. 2012. Weak larval competition between the invasive mosquito, Aedes japonicus japonicus (Diptera: Culicidae) and three resident container inhabiting mosquitoes under standard laboratory conditions. J Med. Entomol. 49:277-285.

Molaei, G., Huang, S., and Andreadis, T. G. 2012. Vector-host interactions of Culex pipienscomplex mosquitoes in northeastern and southern USA. J Amer Mosq. Control Assoc. 28:127-13.

Morningstar, R. J., Hamer, G. L., Goldberg, T. L., Huang, S., Andreadis, T. G., and Walker E. 2012. Diversity of Wolbachia pipientis strain wPip in a genetically admixtured, above- ground Cu/ex pipiens (Diptera: Culicidae) population: association with form molestus ancestry and host selection patterns. J. Med. Entomol. 49:474-481

Simpson, J.E., Hurtado, P. J., Medlock, J. Molaei, G. Andreadis, T. G., Galvani,A. P., and Diuk-Wasser, M.A. 2012. Vector host-feeding preferences drive transmission of multi- host pathogens: West Nile virus as a model system. Proc. R. Soc. B. 279:925-933.

2013

Andreadis, T. G., Takaoka, H., Otsuka, Y., and Vossbrinck, C.R. 2013. Morphological and molecular characterization of a microsporidian parasite, Takaokaspora nipponicus n. gen., n. sp. from the. invasive rock pool mosquito, Ochlerotatusjaponicusjaponicus. J.!nvertebr. Pathol. 114:161-172.

Armstrong, P. M. and Andreadis, T. G. 2013. Eastern equine encephalitis virus: old enemy, new threat. New Engl. J. Med. 368:1670-1673.

20 Armstrong, P. M., Anderson, J. F., Farajollahi, A. Healy, S. P., Unlu, I., Crepeau, T. N. Gaugler, R., Dina M. Fonseca, D. M., and Andreadis, T. G. 2013. Isolations of Cache Valley virus from Aedes albopictus (Diptera: Culicidae) in New Jersey and evaluation of its role as a regional arbovirus vector J. Med. Entomol. 50: 1310-1314.

Huang, S., Smith, V. J., Molaei, G., Andreadis, T. G., Larsen, S. E., and Luccchesi, E. F. 2013. Prevalence of Dirofilaria immitis (Spirurida: Onchocercidae) infection in Aedes, Culex and Culiseta mosquitoes from north San Joaquin Valley, CA. J. Med. Entomol. 50:1315- 1323. Molaei, G., Andreadis, T. G., Armstrong, P. M., Thomas, M. C., Deschamps, T., Cuebas Incle, E., Montgomery, W., Osborne, M., Smale, S., Matton, P., Andrews, W., Best, C., Cornine III, F., Bidlack, E., and Texeira, T. 2013. Vector-host interactions and epizootiology of eastern equine encephalitis virus in Massachusetts, USA. Vector-Borne Zoonotic Dis. 13:312-323.

2014

Nelson, R., Esponda, B. Andreadis, T., Armstrong, P. 2014. West Nile virus -Connecticut, 2000-2013. Connecticut Epidemiologist 34:5-7.

Nelson, R., Ciesielski, T. Andreadis, T., Armstrong, P. 2014. Human case of eastern equine encephalitis - Connecticut, 2013. Connecticut Epidemiologist 34:9-10.

Andreadis, T. G., Anderson, J. F., Armstrong, P. M. and Main, A. J. Spatial-temporal analysis of Cache Valley virus (Bunyaviridae: Orthobunyavirus) infection in Anopheline and Culicine Mosquitoes (Diptera: Culicidae) in the Northeastern United States, 1997 - 2012. Vector-Borne and Zoonotic Dis. (in press).

Becnel, J. J. and Andreadis T. G. 2014. Microsporidia in insects. In: M. Witter and J. J Becnel (eds.), Microsporidia: Pathogens of Opportunity. John Wiley & Sons, Inc., Hoboken, NJ. (in press) Anderson, J. F., Main, A. J., Armstrong, P. E., Andreadis, T. G. and Ferrandino, F. J. Arboviruses in North Dakota, 2003-2006. Am. J. Trop. Med. Hyg. (in press).

APPENDIX H. Effects of Larvacides on Non-Target Organisms

Effects of Larvacides on Non-Target Organisms USFWS 2003

Prepared By

Wennona Brown USFWS, Maryland Coop Unit 1120 Trigg Hall University of Maryland Eastern Shore Princess Anne, MD 21853 (410) 651-7505 FAX (410) 651-7662 e-mail: [email protected]

Introduction

The information contained within this document is a guide to mosquito larvicide effects on nontarget organisms. Included is information on the four most commonly used larvicides: monomolecular surface films (Arosurf7), Bacillus thuringiensis israelensis (BTI), methoprene (Altosid7) and temephos (Abate7). Articles presented are representative entries whose information would lend itself to tabulation. This does not represent a comprehensive treatment of the subject.

The following information is provided for each larvacide: a short description of how the larvicide works, a generalized synopsis of non-target effects, label application rates of various formulations of the product, references cited within the effects table, and a tabulation of nontarget effects on various organisms. The table is arranged by taxonomic categories (e.g., birds, insects), and alphabetical within category. Taxonomy may not be the most current. Label application rates are excerpted from the manufacturer=s information sheets as follows: Arosurf7 now manufactured as Agnique7 by the Henkel Corporation; BTI (Vectobac7 products) manufactured by Abbott Laboratories; Methoprene (Altosid7 products) manufactured by Sandoz Agro, Inc., and Zoecon; Temephos (Abate7 products) manufactured by Clarke Mosquito Control Products, Inc. At the end of each section is an extensive bibliography of mosquito larvicide articles. Additional information is available on the Internet. Two web sites that are useful include:

http://ace.ace.orst.edu/info/extoxnet/pips/ghindex.html These Pesticide Information Profiles provide general information on many registered pesticides, such as mode of action, toxicity, ecological effects, and references.

http://www.famu.edu/jamsrl/peis/mosquito/mosqsearch.html The Non-target Search Form provides a searchable database for mosquito literature. A search can be conducted by author, organism, pesticide, or a key word search. The database provides abstracts for many of the articles.

Outline

Arosurf7 (ISA-20E) Synopsis of Non-target Effects Label Application Rates for Agnique References Cited Table 1. Non-target Effects of Arosurf7 Fish Mollusks Crustaceans Insects Annelids Bacillus thuringiensis israelensis (BTI) Synopsis of Non-target Effects Label Application Rates References Cited Table 2. Non-target Effects of BTI Fish Crustaceans Insects Methoprene Synopsis of Non-target Effects Label Application Rates References Cited Table 3. Non-target Effects of Methoprene Fish Amphibians Arachnids Mollusks Crustaceans Insects Annelids Aschelminths Flatworms Protozoa Phytoplankton Temephos Synopsis of Non-target Effects Label Application Rate

References Cited Table 4. Non-target Effects of Temephos Birds Reptiles Amphibians Fish Arachnids Mollusks Crustaceans Insects Annelids Aschelminths Flatworms Plankton

Mosquito Bibliography

7 Arosurf (ISA-20E) now produced as Agnique7

Arosurf is a monomolecular surface film, which reduces the water surface tension. This interferes with larval orientation at the air-water interface and/or increases wetting tracheal structures, thus suffocating the organism. As the film spreads over the water surface, it tends to concentrate the larvae, which may increase mortality from crowding stress (Dale and Hulsman 1990).

According to the Henkel Corporation, Agnique=s improvements over Arosurf center around removal of the byproducts that left the white residues in the drums and application equipment. Removing these byproducts lowered the freezing point of the product. The spreading ability was also improved, so that application of the product was made easier.

Synopsis of Non-target Effects

Arosurf had no adverse effect on any of the organisms tested. However, none of the studies listed investigated species such as water boatman or backswimmers.

Label Application Rates for Agnique

Example habitat: Salt-marsh, ponds, storm water retention basins, roadside ditches, grassy swales, potholes, fields, reservoirs, irrigated croplands, etc. Larvae: 0.2-0.5 gal/surface acre Pupae: 0.2-0.3 gal/surface acre

Example habitat: Pumping station bunkers, settings, polishing and evapo-percolation ponds of sewage treatment systems, drainage areas containing effluent from slaughter houses, etc. Larvae: 0.4-0.5 gal/surface acre Pupae 0.2-0.3 gal/surface acre

References Cited:

Dale, P.E.R. and K. Hulsman. 1990. A critical review of salt marsh management methods for mosquito control. Review in Aquatic Sciences 3:281-311.

Hester, P.G., M.A. Olson, and J.C. Dukes. 1991. Effects of ArosurfR MSF on a variety of aquatic nontarget organisms in the laboratory. J. Amer. Mosq. Control Assn. 7:48-51.

Mulla, M.S., H.A. Darwazeh, and L.L. Luna. 1983. Monolayer films as mosquito control agents and their effects on non-target organisms. Mosq. News. 43:489- 495.

J-5

Table 1. Non-target Effects of Arosurf7 (now produced as Agnique7) Classification Organism Reference Formulation Application Adverse No Comments Rate (study) Effects Effects

Longnose Hester et MSF X 96-hour acute static toxicity lab test Fish 47 ml/m2 killifish al.1991 Atheriniformes (50 (Fundulus gal/acre) simulus)

Snail (Physa sp.) Hester et MSF X 96-hour acute static toxicity lab test Mollusks 47 ml/m2 al.1991 Basommatophora (50 gal/acre) Amphipod Hester et MSF X 96-hour acute static toxicity lab test Crustaceans 47 ml/m2 (Grammarus al.1991 Amphipoda (50 spp.& unknown) gal/acre) Anostraca Fairy shrimp Hester et MSF X 96-hour acute static toxicity lab test 47 ml/m2 (Streptocephalus al.1991 (50 seali) gal/acre) Copepoda Copepods Mulla et al MSF 0.5-0.75 X information from abstract 1983 gal/acre

Decapoda Fiddler crab Hester et MSF X 96-hour acute static toxicity lab test 47 ml/m2 (Uca spp.) al.1991 (3.3% mortality, not attributed to test) (50 gal/acre) Decapoda Freshwater shrimp Hester et MSF X 96-hour acute static toxicity lab test 47 ml/m2 (Palaemonetes al.1991 (50 gal/acre)

paludosus) Decapoda Grass shrimp Hester et MSF X 96-hour acute static toxicity lab test 47 ml/m2 (Palaemonetes al.1991 (50 pugio) gal/acre) Decapoda Crayfish Hester et MSF X 96-hour acute static toxicity lab test 47 ml/m2 (Procambarius al.1991 (50 spp.) gal/acre) Isopoda Isopod (Asellus Hester et MSF X 96-hour acute static toxicity lab test 47 ml/m2 spp.) al.1991 (50 gal/acre) Ostracoda Seed shrimp Mulla et al MSF 0.5-0.75 X information from abstract 1983 gal/acre

Insects Diving Beetle Mulla et al MSF 0.5-0.75 X information from abstract Coleoptera adults (Berosus 1983 gal/acre metalliceps) Ephemeroptera Mayfly naiads Mulla et al MSF 0.5-0.75 X information from abstract (Callibaetis 1983 gal/acre pacificus)

Polychaete Hester et MSF X 96-hour acute static toxicity lab test Annelids 47 ml/m2 (Laeonereis al.1991 Polychaeta (50 culveri) gal/acre)

Bacillus thuringiensis israelensis (BTI)

BTI is a bacterial pathogen which produces a parasporal body know as a Acrystal.@ This toxin kills larvae rapidly by attacking the plasma membrane of the gut epithelia (Dale and Hulsman, 1990). BTI forms asexual reproductive spores that enable it to survive adverse conditions; during spore formation, BTI produces unique crystalline bodies as a companion product. These spores and crystals must be ingested before they act as poisons to target insects (referred to as a Astomach@ poison). The crystals dissolve in response to intestinal conditions of susceptible insect larvae. The toxins released paralyze the gut, thus interfering with normal digestion which triggers the insect to stop feeding. Then the BTI spores can invade other tissues and multiply in the bloodstream until the insect dies. BTI is ineffective against adult insects. BTI is effective against mosquitoes, black flies, and certain midges. Other strains of Bacillus thuringiensis are effective against other insects, such as the wax moth, gypsy moth and cabbage looper, and a new strain has been found is effective against the boll weevil (Pesticide Information Profile, EXTOXNET).

Synopsis of Non-target Effects

The attached Table 2 presents detailed information regarding the effects of BTI on non-target organisms. A few generalizations can be drawn from this information. Target organisms for BTI applications are various species of mosquitoes (both freshwater and salt marsh) and black flies. Effectiveness of BTI on mosquito species is not included. Chironomids, also a Dipteran (like mosquitoes and black flies), were primarily the non-target group adversely affected by BTI, but this also varied by species. A 3-year study found the other Dipterans (crane flies and stone flies) were affected in the third year of the study, as were the Ephemeropterans (mayflies).

Label Application Rates

Vectobac G (200 International Toxic Units/mg = 0.091 billion ITU/lb) Habitat: irrigation/roadside ditches, floodwater, standing ponds, woodland pools, catch basins, storm water retention ponds, tidal water, and salt marshes Application Rate: 2.5 - 10 lbs/acre

Late 3rd instar or early 4th instar, high populations, or heavily polluted water (sewage lagoons, etc.) or abundant algae Application Rate: 10 - 20 lbs/acre.

Allow 7 to 14 days between applications.

Vectobac 12AS (1200 ITU/mg = 4.84 billion ITU/gal or 1.279 billion ITU/liter) Mosquito habitat: irrigation/roadside ditches, floodwater, standing ponds, woodland pools, catch basins, storm water retention ponds, tidal water, salt marshes, rice fields Application rate: 0.25 - 1 pt/acre

Use higher rate in polluted water and when late 3rd and early 4th instar larvae predominate, when mosquito population is high, water is heavily polluted, or abundant algae Application rate: 1- 2 pts/acre

Blackfly habitat B streams stream water (=ppm) for 1 minute exposure time: 0.5 - 25mg/liter stream water (=ppm) for 10 minute exposure time: 0.05 - 2.5 mg/liter (use higher rate range when stream contains high concentration of organic materials, algae or dense aquatic vegetation)

Vectobac CG (200 ITU/mg = 0.091 billion ITU/lb) Habitat: irrigation/roadside ditches, floodwater, standing ponds, woodland pools, catch basins, storm water retention ponds, tidal water, salt marshes, rice fields Application rate: 2.5 - 10 lb/acre Allow 7

to 14 days between applications.

Web Site:

Pesticide Information Profiles, EXTOXNET http://ace.ace.orst.edu/info/extoxnet/pips/ghindex.html

References Cited:

Charbonneau, C.S., R.D. Drobney, and C.F. Rabeni. 1994. Effects of Bacillus thuringiensis var. Israelensis on nontarget benthic organisms in a lentic habitat and factors affecting the efficacy of the larvicide. Environ. Tox. Chem. 13:267-279.

Cilek, J.E. and F.W. Knapp. 1992. Distribution and control of Chironomus riparius (Diptera: Chironomidae) in a polluted creek. J. Amer. Mosq. Control Assn. 8:181-183.

Colbo, M.H. and A.H. Undeen. 1980. Effect of Bacillus thuringiensis var. israelensis on nontarget insects in stream trials for control of Simuliidae. Mosq. News 40:368-371.

Dale, P.E.R. and K. Hulsman. 1990. A critical review of salt marsh management methods for mosquito control. Review in Aquatic Sciences 3:281-311. Hershey, A.E., A.R. Lima, G.J. Niemi, and R.R. Regal. 1998. Effects of Bacillus thuringiensis israelensis (BTI) and methoprene on nontarget macroinvertebrates in Minnesota wetlands. Ecol. Appl. 8:41-60.

Knepper, R.G. and E.D. Walker. 1989. Effect of Bacillus thuringiensis israelensis (H-14) on the isopod Asellus forbesi and the spring Aedes mosquitoes in Michigan. J. Am Mosq. Control Assoc. 5:596-598.

Lee, B.M. and G.I. Scott. 1989. Acute toxicity of temephos, fenoxycarb, diflubenzuron, and methoprene and Bacillus thuringiensis var. israelensis to the Mummichog (Fundulus heteroclitus). Bull. Environ. Contamin. And Toxicol. 43:827-832.

Miura, T., R.M. Takahashi, and F.S. Mulligan, III. 1980. Effects of the bacterial mosquito larvicide, Bacillus thuringiensis serotype 14 on selected aquatic organisms. Mosq. News 40:619-622.

Molloy, D.P. 1992. Impact of the black fly (Diptera: Simuliidae) control agent Bacillus thuringienses var. Israelensis on chironomids (Diptera: Chironomidae) and other nontarget insects: results of ten field trials. J. Amer. Mosq. Control Assn. 8:24-31.

Molloy, D. And H. Jamnback. 1981. Factors influencing efficacy of Bacillus thuringiensis var. israelensis as a blackfly biocontrol agent and its effect on nontarget stream insects. J. Econ. Entomol. 74:314-318.

Mulligan, F.S., III and C.H. Schaefer. 1981. Integration of a selective mosquito control agent Bacillus thuringiensis Serotype H-14, with natural predator populations in pesticide-sensitive habitats. Proceedings California Mosquito Vector Control Association 49:19-22. Rodcharoen, J., M.S. Mulla and J.D. Chaney. 1991. Microbial larvicides for the control of nuisance aquatic midges (Diptera: Chironomidae) inhabiting mesocosms and man-made lakes in California. J. Amer. Mosq. Control Assn. 7:56-62.

Table 2. Non-target Effects of BTI

Classification Organism Reference Formulation Application Rate Adverse No Comments (study) Effects Effects

Mummichog Lee & Scott Vectobac EC 96-hour LC = 980 mg/L Fish 50 (Fundulus 1989 Atheriniformes (1,176,000 ITU/L); no effect conc. heteroclitus) =22.36 mg/L

Cypriniformes Golden Shiner Mulligan & B.t. H-14 1.0 kg/ha & X hand applied to bait fish ponds (Notemigonus Schaefer (692 69 2.0 kg/ha crysoleucas) 1981 ITU/mg)

Water fleas Miura et al SAN 402 I X experimental plots Crustaceans 0.25 kg/ha 1980 WDC Cladocera (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Cladocera Water fleas Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands Schaefer (552 22 1981 ITU/mg)

Conchostraca Clam Shrimp Miura et al SAN 402 I X experimental plots 0.25 kg/ha 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml)

Copepoda Copepods Miura et al SAN 402 I 0.25 kg/ha X experimental plots 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml)

Copepoda Eucopepoda Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands Schaefer (552 22 1981 ITU/mg) Isopoda Asellus forbesi Knepper & Bti X hardwood bottomland pools; isopods Walker 1989 not negatively affected (information from abstract)

Ostracoda Podocopa Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands Schaefer (552 22 1981 ITU/mg)

Ostracoda Seed shrimp Miura et al SAN 402 I X experimental plots 0.25 kg/ha 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Beetles Colbo & B.t. H-14 X flowing stream Insects 1x105spores/ml Undeen 1980 Coleoptera

Coleoptera Beetles Mulligan & B.t. H-14 1.1 kg/ha X aerially applied to duck club pond Schaefer (576 20 1981 ITU/mg)

Coleoptera Beetles Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands Schaefer (552 22 1981 ITU/mg)

Coleoptera Beetles Hershey et Vectobac G 11.720.64 natural wetlands; aerial application, al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); significantly reduced in 1993 season only; not significantly reduced over the 3- yr period

Coleoptera Dytiscid beetles Miura et al SAN 402 I X experimental plots 0.25 kg/ha 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Coleoptera Elmids Molloy & Primary powder X 0.5ppm conc. stream study, flow rate 1770 l/min; Jamnback (R153-78) 8 o o 1981 (1.4x10 water temp. range 8 - 17 C spores/mg) Coleoptera Hydrophilid Miura et al SAN 402 I X experimental plots 0.25 kg/ha beetles 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Diptera Biting midges Hershey et Vectobac G 11.720.64 X natural wetlands; aerial

Ceratopogonids al. 1998 kg/ha application, 6 treatments per year for 3 years (1991-1993 long-term effects study); not affected until 3rd year, reduced by 67% in 1993; reduced by 29% over the 3-yr period

Diptera Black flies Molloy & Primary powder 0.5ppm conc. X stream study, flow rate 1770 l/min; Jamnback (R153-78) (1.4x108 water temp. range 8o- 17oC 1981 spores/mg) Diptera Black flies Molloy 1992 Bactimos WP ranged from X flowing streams; water temp. ranged Teknar WDC 3.7ppm/15 min o o Vectobac WP to 50ppm/1min from 3 C to 17 C; discharge rates ranged from 168 l/min to 20,740 l/min

Diptera Black flies Colbo & B.t. H-14 1x105spores/ml X flowing stream (Simuliidae) Undeen 1980 Diptera Chironomid: Rodcharoen Vectobac 6253 56 kg/ha X lake study; no effect Procladius et al. 1991 (200 ITU/mg) bellus & corn grit Tanypus granules neopunctipenni s

Diptera Chironomid: Rodcharoen Vectobac 6253 56 kg/ha X lake study; 42-67% control for 3 Chironomus et al. 1991 (200 ITU/mg) weeks decorus corn grit

granules Diptera Chironomid: Rodcharoen Vectobac ABG 1.4 kg/ha X lake study; no effect noted Procladius et al. 1991 6164 (technical 2.8 kg/ha bellus & powder) Tanypus neopunctipenni s

Diptera Chironomid: Rodcharoen Vectobac technical 2.2 kg/ha X lake study; lower rate yielded Chironomus et al. 1991 powder (5,000 4.5 kg/ha maximum control of 66% at 2 decorus ITU/mg) 6.7 kg/ha weeks; middle rate yielded higher level of control; higher rate yielded 95% control at 1 week, then 100% at 2-3 weeks; higher 2 rates yielded over 70% control for 4 weeks

Diptera Chironomid: Rodcharoen Vectobac 6264 11.2 kg/ha lake study: Amediocre@ control (32 Chironomus et al. 1991 (400 ITU/mg) 19.1 kg/ha & 47% respectively) for decorus corn grit about 2 weeks granules

Diptera Chironomid: Rodcharoen Vectobac 6253 13.5 kg/ha X lake study; unaffected even at highest Procladius et al. 1991 (200 ITU/mg) 28 kg/ha rate bellus & corn grit 56kg/ha Tanypus granules grodhausi

Diptera Chironomid: Rodcharoen Vectobac 6264 11.2 and 22.4 X mesocosm studies; highly Dicrotendipes et al. 1991 (400 ITU/mg) kg/ha susceptible sp. corn grit granules Diptera Chironomid: Rodcharoen Vectobac ABG 1.4 kg/ha X lake study; lower rate yielded Chironomus et al. 1991 6164 (technical 2.8 kg/ha maximum reduction of 73% 2 weeks decorus powder) post treatment and lasted about 4 weeks; high rate yielded max. control of 87% at 3 weeks post treatment

Diptera Chironomid: Rodcharoen Vectobac 6253 22.4 and 44.8 X mesocosm studies; highly Chironomus sp. et al. 1991 (200 ITU/mg) kg/ha susceptible to higher rate corn grit granules

Diptera Chironomid: Rodcharoen Vectobac 6 AS 11.2 and 22.4 X mesocosm studies; 11.2 kg/ha Chironomus sp. et al. 1991 (aqueous kg/ha yielded 37% control at 1 week post suspension) treatment; 22.4 kg/ha yielded 57% control at 2 weeks post treatment; conclusion that control not evident until 2 weeks post treatment

Diptera Chironomid: Rodcharoen Vectobac 6264 11.2 and 22.4 X mesocosm studies; lower rate yielded Procladius sp. et al. 1991 (400 ITU/mg) kg/ha 24% control after 1 week; higher rate corn grit no effect; conclusion little to no granules control

Diptera Chironomid: Rodcharoen Vectobac ABG- 5.6 and 11.2 X mesocosm studies; yielded 98% Chironomus sp. et al. 1991 6164 (technical kg/ha and 100% control (respectively) at powder) 2 weeks post treatment

Diptera Chironomid: Rodcharoen Vectobac 6264 11.2 and 22.4 X mesocosm studies; conclusion little Paratanytarsus et al. 1991 (400 ITU/mg) kg/ha to no control sp. corn grit granules

Diptera Chironomid: Rodcharoen Vectobac 6253 22.4 and 44.8 X mesocosm studies; conclusion little Paratanytarsus et al. 1991 (200 ITU/mg) kg/ha to no control sp. corn grit granules

Diptera Chironomid: Rodcharoen Vectobac 6253 13.5 kg/ha X lake study; lowest rate showed only Chironomus et al. 1991 (200 ITU/mg) 28 kg/ha 22% control at 2 weeks; higher rates decorus corn grit 56kg/ha showed 83% and 96% control granules (respectively); control of over 70% at 2 higher rates lasted over 4 weeks

Diptera Chironomid: Rodcharoen Vectobac 6253 22.4 and 44.8 X mesocosm studies; highly Dicrotendipes et al. 1991 (200 ITU/mg) kg/ha susceptible sp. corn grit granules

Diptera Chironomid: Rodcharoen Vectobac 6264 11.2 and 22.4 X mesocosm studies; highly Chironomus sp. et al. 1991 (400 ITU/mg) kg/ha susceptible corn grit granules Diptera Chironomid: Rodcharoen Vectobac 6253 22.4 and 44.8 X mesocosm studies; lower rate no Procladius sp. et al. 1991 (200 ITU/mg) kg/ha effect; higher rate yielded 17% corn grit control after 1 week conclusion little granules to no control

Diptera Chironomidae Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); reduced by 66% in 1992 & 84% in 1993

Diptera Chironomids Cilek & Vectobac-6AS 50 ppm X field test in flowing creek, velocity Knapp 1992 0.8 m/s & 0.5 m/s, water temp. 25oC, pH 7.5 Diptera Chironomids Cilek & Vectobac-G 22.4 kg/ha X field test in flowing creek, velocity Knapp 1992 0.1 m/s, water temp. 25oC, pH 7.5

Diptera Chironomids Mulligan & B.t. H-14 0.8 kg/ha aerially applied to wetlands; peak Schaefer (552 22 numbers 1 day after treatment, with 1981 ITU/mg) gradual decline thereafter

Diptera Chironomids Molloy & Primary powder X 0.5ppm conc. stream study, flow rate 1770 l/min; Jamnback (R153-78) (1.4x108 water temp. range 8o- 17oC

1981 spores/mg) Diptera Chironomids Miura et al SAN 402 I X experimental plots; all larvae 0.25 kg/ha 1980 WDC 3 collected were killed w/i 2 days of (~1.3x10 treatment, but daily collections spores/ml) & 1 3 rapidly increased indicating short- kg/ha (~5.4x10 term effects spores/ml) Diptera Chironomids Molloy 1992 Bactimos WP ranged from X flowing streams; water temp. ranged (filter-feeders; Teknar WDC 3.7ppm/15 min o o Rheotanytarsus Vectobac WP to 50ppm/1min from 3 C to 17 C; discharge rates spp.) ranged from 168 l/min to 20,740 l/min Diptera Molloy 1992 Bactimos WP ranged from X Chironomids B flowing streams; water temp. ranged Teknar WDC 3.7ppm/15 min Other o o Vectobac WP to 50ppm/1min from 3 C to 17 C; discharge rates ranged from 168 l/min to 20,740 l/min Diptera Chironomids Colbo & B.t. H-14 X flowing stream 1x105spores/ml Undeen 1980

Diptera Chironomids Charbonneau Vectobac-G 28.1 kg/ha X adversely affected in lab, but et at 1994 environmental factors (temperature, larval instar, water depth & water surface area coverage) reduced efficacy in the field

Diptera Chironomids Charbonneau Vectobac-G 5.6 kg/ha X adversely affected in lab, but et al 1994 environmental factors

(temperature, larval instar, water depth & water surface area coverage) reduced efficacy in the field

Diptera Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, Chironomids B al. 1998 kg/ha 6 treatments per year for 3 years predatory (1991-1993 long-term effects study); reduced 62% in 1992 & 83% in 1993

Diptera Crane flies Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, (Tipulidae) al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); reduced by 73% over the 3-yr treatment period

Diptera Diptera Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); over 3-yr study total reduction = 63%

Diptera Nematocera Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, 6 al. 1998 kg/ha treatments per year for 3 years (1991- 1993 long-term effects study); over 3- yr treatment total reduction = 67%

Diptera Soldier flies Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, (Stratiomyidae) al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); reduced in 1993 season only, reduction =74%; yielding 56% reduction over the 3-yr period

Ephemeroptera May flies Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, (Brachycera) al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); showed no effect until 1993 when reduced by 66%

Ephemeroptera Mayflies Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands Schaefer (552 22 1981 ITU/mg)

Ephemeroptera Mayflies Miura et al SAN 402 I X experimental plots 0.25 kg/ha 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Ephemeroptera Mayflies Molloy 1992 Bactimos WP ranged from X flowing streams; water temp. ranged Teknar WDC 3.7ppm/15 min o o Vectobac WP to 50ppm/1min from 3 C to 17 C; discharge rates ranged from 168 l/min to 20,740 l/min

Ephemeroptera Mayflies Molloy & Primary powder 0.5ppm conc. X stream study, flow rate 1770 l/min; Jamnback (R153-78) (1.4x108 water temp. range 8o- 17oC 1981 spores/mg) Ephemeroptera Mayflies Colbo & B.t. H-14 X flowing stream 1x105spores/ml Undeen 1980

Hemiptera Corixids Miura et al SAN 402 I 0.25 kg/ha X experimental plots 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Hemiptera Notonectids Miura et al SAN 402 I X experimental plots 0.25 kg/ha 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Hemiptera True bugs Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands Schaefer (552 22 1981 ITU/mg)

Hemiptera True bugs Mulligan & B.t. H-14 1.1 kg/ha X aerially applied to duck club pond Schaefer (576 20 1981 ITU/mg)

Odonata Damselflies Miura et al SAN 402 I 0.25 kg/ha X experimental plots 1980 WDC (~1.3x103 spores/ml) & 1

kg/ha (~5.4x103 spores/ml) Odonata Damselflies Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands (Zygoptera) Schaefer (552 22 1981 ITU/mg)

Odonata Dragonflies & Colbo & B.t. H-14 1x105spores/ml X flowing stream Damselflies Undeen 1980 Odonata Dragonflies Mulligan & B.t. H-14 0.8 kg/ha X aerially applied to wetlands (Anisoptera) Schaefer (552 22 1981 ITU/mg)

Odonata Dragonflies Mulligan & B.t. H-14 1.1 kg/ha X aerially applied to duck club pond (Anisoptera) Schaefer (576 20 1981 ITU/mg)

Odonata Dragonflies Miura et al SAN 402 I X experimental plots 0.25 kg/ha 1980 WDC (~1.3x103 spores/ml) & 1 kg/ha (~5.4x103 spores/ml) Plecoptera Stoneflies Colbo & B.t. H-14 X flowing stream 1x105spores/ml Undeen 1980

Plecoptera Stoneflies Molloy & Primary powder 0.5ppm conc. X stream study, flow rate 1770 l/min; Jamnback (R153-78) (1.4x108 water temp. range 8o- 17oC 1981 spores/mg)

Trichoptera Caddisflies Colbo & B.t. H-14 X flowing stream 1x105spores/ml Undeen 1980

Trichoptera Caddisflies Molloy & Primary powder X 0.5ppm conc. stream study, flow rate 1770 l/min; Jamnback (R153-78) 8 o o 1981 (1.4x10 water temp. range 8 - 17 C spores/mg) Trichoptera Caddisflies Molloy 1992 Bactimos WP ranged from X flowing streams; water temp. ranged Teknar WDC 3.7ppm/15 min o o Vectobac WP to 50ppm/1min from 3 C to 17 C; discharge rates ranged from 168 l/min to 20,740 l/min. Miscellaneous Non-dipteran Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, predators al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); no significant seasonal effect in 1991 & 1992, but significant reduction in 1993

Miscellaneous Total predatory Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, insects al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term effects study); no significant seasonal effect in 1991 & 1992, but 60% reduction in 1993

Miscellaneous Non-insect macro- Hershey et Vectobac G 11.720.64 X natural wetlands; aerial application, invertebrates al. 1998 kg/ha 6 treatments per year for 3 years (1991-1993 long-term

effects study)

Methoprene

Methoprene is an insect growth regulator (IGR), which mimics juvenile hormones (Dale and Hulsman 1990). It interferes with the insect=s maturation stages and makes it impossible for the insect to reach the adult stage, thus preventing it from reproducing. Methoprene is considered a biochemical pesticide because it interferes with the life cycle rather than direct toxicity. To be effective, it must be administered at the proper life stage of the mosquito (or target species). It is not toxic to pupal or adult stages. Treated larvae will pupate, but will not emerge as adults (Pesticide Information Profiles, EXTOXNET).

Synopsis of Non-target Effects

As seen in Table 3, methoprene had no effect on the vertebrate species tested. Mixed effects were seen for snails, and crustaceans such as grass shrimp and mud crabs. Insects most affected were dipterans, with some mixed effects reported for mayflies and some coleopterans.

Label Application Rates

Altosid7 Liquid Larvicide (A.L.L.) effective on 2nd, 3rd, or 4th instar larvae of floodwater mosquitoes; has no effect on pupae or adult mosquitoes

Crop Areas: irrigated croplands after flooding, e.g. vineyards, rice fields, irrigated pastures, berry fields, orchards, bogs. Application Rate: 3 to 4 fluid ounces/acre (219 to 293 ml/hectare) in water.

Intermittently Flooded Areas: freshwater swamps and marshes, salt marshes, woodland pools and meadows, dredging spoil sites, drainage areas, waste treatment and settling ponds, ditches and other natural or man-made depressions. Application Rate: 3 to 4 fluid ounces/acre (219 to 293 ml/hectare) in water.

Dense Vegetation or Canopy Areas: Apply A.L.L. on sand granules at standard application rate (as stated above).

Altosid7 Pellets is toxic to aquatic dipteran (mosquitoes) and Chironomid (midge) larvae. It has no effect on pupal or adult stage mosquitoes; pellets release effective levels for up to 30 days.

Floodwater Sites: pastures, meadows, rice fields, freshwater swamps and marshes, salt and tidal marshes, cattail marshes, woodland pools, floodplains, tires, and other artificial water-holding containers. Application Rate: 2.5 - 5.0 lb/acre.

Floodwater Sites: dredging spoil sites, waste treatment and settling ponds, ditches and other man-made depressions. Application Rate: 5.0 - 10.0 lb/acre.

Permanent Water Sites: ornamental ponds and fountains, fish ponds, cattail marshes, water hyacinth beds, flooded crypts, transformer vaults, swimming pools and other man-made depressions, etc. Application Rate: 2.5 - 5.0 lb/acre

Permanent Water Sites: storms drains, catch basins, roadside ditches, cesspools, septic tanks, waste settling ponds, vegetation-choked phosphate pits. Application Rate: 5.0 - 10.0 lb/acre.

Altosid7 XR-G (extended residual granules) is toxic to aquatic dipteran; it has no effect on pupal or adult life stages; length of control up to 21 days, but actual length depends on duration and frequency of flooding.

Non-Crop Areas: snow pools, salt and tidal marshes, freshwater swamps and marshes, woodland pools and meadows, dredging spoil sites, drainage areas, ditches, water-holding receptacles and other natural or man-made depressions.

Aedes, Anopheles, and Psorophora spp. Application Rate: 5 - 10 lb/acre (5.6 - 11.2 kg/ha). Culex, Culiseta, Coquillettidia, and Mansonia spp. Application Rate: 10 - 20 lb/acre (11.2 - 22.4 kg/ha). Within these rates, use lower rate when water is shallow (2 ft. [60 cm]) and vegetation and/or pollution are minimal. Use higher rates when water is deep (2 feet) and vegetation and/or pollution are heavy.

Altosid7 Briquets: toxic to aquatic dipterans; no effect on pupal or adult stage mosquitoes; under normal conditions, repeat treatment every 30 days; designed to control mosquitoes in small bodies of water.

Sites: storm drains, catch basins, roadside ditches, fish ponds, ornamental ponds and fountains, septic tanks, waste treatment and settling ponds, abandoned swimming pools, other man-made depressions, cattail marshes, water hyacinth beds, pastures, meadows, rice fields, freshwater swamps and marshes, salt and tidal marshes, woodland pools, floodplains, dredging spoil sites.

Application Rates: non-(or low) flow, shallow depressions (up to 2 ft. deep), treat on basis of surface area placing one briquet per 100 sq. ft. Flowing water or deeper than 2 ft, treat on basis of volume, one briquet per 10 cu ft. (75 gal of water).

Altosid7 XR (extended residual briquets): toxic to aquatic dipterans; no effect of pupal or adult stage mosquitoes; one application should last entire breeding season, or 150 days.

Sites: storm drains, catch basins, roadside ditches, fish ponds, waste treatment and settling ponds, cattail marshes, meadows, rice fields, freshwater swamps and marshes, salt and tidal marshes, woodland pools, floodplains and dredging spoil sites.

Application Rates: Aedes and Psorophora spp. in non-(or low) flow shallow depressions (2 ft. deep) treat on basis of surface area - 1 briquet per 200 ft2. Culex, Culiseta, and Anopheles spp. - 1 briquet per 100 ft2. Coquillettidia and Mansonia spp. for application to cattail marshes and water hyacinth beds, place 1 briquet per 100 ft.2.

Web Site:

Pesticide Information Profiles, EXTOXNET http://ace.ace.orst.edu/info/extoxnet/pips/ghindex.html

References Cited:

Ali, A. 1991. Activity of new formulations of methoprene against midges (Diptera: Chironomidae) in experimental ponds. J. Amer. Mosq. Control Assn. 7:616-620.

Breaud, T.P., J.E. Farlow, C.D. Steelman and P.E. Schilling. 1977. Effects of the insect growth regulator methoprene on natural populations of aquatic organisms in Louisiana intermediate marsh habitats. Mosq. News 37:704-712.

Celestial, D.M. and C.L. McKenney, Jr. 1994. The influence of an insect growth regulator on the larval development of the mud crab Rhithropanopeus harrisii. Environ. Poll. 85:169-173.

Dale, P.E.R. and K. Hulsman. 1990. A critical review of salt marsh management methods for mosquito control. Review in Aquatic Sciences 3:281-311.

Ellgaard, E.G., J.T. Barber, S.C. Tiwari and A.L. Friend. 1979. An analysis of the swimming behavior of fish exposed to the insect growth regulators, methoprene and diflubenzuron. Mosq. News 39:311-314.

Hershey, A.E., A.R. Lima, G.J. Niemi, and R.R. Regal. 1998. Effects of Bacillus thuringiensis israelensis (BTI) and methoprene on nontarget macroinvertebrates in Minnesota wetlands. Ecol. Appl. 8:41-60.

McAlonan, W.G., F.J. Murphey and R.W. Lake. 1976. Effects of two insect growth regulators on some selected saltmarsh non-target organisms. Proc. Ann. Meet. N.J. Mosq. Control Assoc. 63:198.

McKenney, C.L., Jr. and D.M. Celestial. 1996. Modified survival, growth, and reproduction in an estuarine mysid (Myosidposis bahia) exposed to a juvenile hormone analogue through a complete life cycle. Aquatic Toxicol. 35:11-20.

McKenney, C.L., Jr. and E. Matthews. 1990. Influence of an insect growth regulator on the larval development of an estuarine shrimp. Environ. Poll. 64:169-178.

Miura, T. And R.M. Takahashi. 1973. Insect development inhibitors. 3. Effects on nontarget aquatic organisms. J. Econ. Entomol. 66:917-922.

Norland, R.L. and M.S. Mulla. 1975. Impact of Altosid on selected members of an aquatic ecosystem. Environ. Entomol. 4:145-152.

Quistad, G.B., D.A. Schooley, L.E. Staiger, B.J. Bergot, B.H. Sleight, and K.J. Macek. 1976. Environmental degradation of the insect growth regulator methoprene. IX. Metabolism by bluegill fish. Pest. Biochem. Physiol. 6:523-529.

Table 3. Non-target Effects of Methoprene

Classification Organism Reference Formulation Application Adverse No Comments Rate (study) Effects Effects

Killifish McAlonan et Altosid 10- 0.012 to 0.120 X caused no mortality Fish at. 1976 F lbs AI/A Atheriniformes Atheriniformes Mosquitofish Ellgaard et methoprene 0.2 ppm X exposed for 12 days; methoprene was (Gambusia affinis) al 1979 added at rate every 2 days such that total conc was increased by 0.1 ppm; no effect on motility

Atheriniformes Mosquitofish Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Gambusia affinis) 1977

Atheriniformes Mosquitofish Miura & technical 5 laboratory toxicity tests: 0% mortality (Gambusia affinis) Takahashi ZR-515 concentrations at 1 ppm; 60% at 100 ppm; test 1973 duration 312 hours Atheriniformes Mummichog Lee & Scott methoprene 96-hour LC = 124.95 mg/L; no (Fundulus 1989 EC 50 heteroclitus) effect concentration = 24.68 mg/L

Cypriniformes Goldfish Ellgaard et methoprene 0.2 ppm X exposed for 13 days; methoprene was al 1979 added at rate every 2 days such that total conc was increased by 0.1 ppm; no effect on motility

Cypriniformes Heterandria Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18

formosa 1977 months; Perciformes Bluegill Quistad et methoprene 0.31 & 0.005 radioactive tag to study uptake; al. 1976 ppm higher dose, fish exhibited stress (LC50 = 2.1 ppm); within 2 weeks after treatment, 93-95% residue had been eliminated

Western toad Miura & technical 5 laboratory toxicity tests: 0% Amphibians tadpoles, Bufo Takahashi ZR-515 concentrations mortality at 1 ppm; test duration 24 Anura borcas helophilus 1973 hours

Oribateid mites Miura & technical X irrigated pasture study Arachnids Takahashi ZR-515 Acarina 1973

Physa sp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; Mollusks 1977 Basommatophora Basommatophora Pond snail, Physa Miura & technical 5 X laboratory toxicity tests: 0% spp. Takahashi ZR-515 concentrations mortality at 100 ppm; test duration 1973 72 hours

Basommatophora Snail, Lymnaea sp. Miura & technical 5 X laboratory toxicity tests: 0% Takahashi ZR-515 concentrations mortality at 1 ppm; test duration 72 1973 hours

Crustaceans Hyallela azteca Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; Amphipoda (Scud) 1977 greater reduction in open water habitats

Amphipoda Sideswimmers, Miura & technical 5 laboratory acute toxicity tests: Hyallela azteca Takahashi ZR-515 concentrations LC50 = 1.25 ppm; test duration 1973 24-120 hours

Cladocera Water fleas, Miura & technical 5 laboratory acute toxicity tests: LC50 Daphnia magna Takahashi ZR-515 concentrations = 0.90 ppm; test duration 24 hours 1973

Cladocera Water fleas, Miura & technical X irrigated pasture study Daphnia magna Takahashi ZR-515 1973

Cladocera Water fleas, Miura & technical 0.7 lb corncob X outdoor cage study Daphnia magna Takahashi ZR-515 granular/acre 1973

Cladocera Water fleas, Miura & technical 0.1 ppm X outdoor test in artificial container; no Daphnia magna Takahashi ZR-515, detectable effects 1973 10% flowable liquid (slow release)

Conchostraca Clam shrimp, Miura & technical 0.1 lb EC/acre X outdoor caged study Eulimnadia sp. Takahashi ZR-515 1973

Conchostraca Clam shrimp, Miura & technical 5 laboratory acute toxicity tests: LC50 Eulimnadia sp. Takahashi ZR-515 concentrations = 1.00 ppm; test duration 24 hours 1973

Conchostraca Clam shrimp, Miura & technical 0.7 lb corncob X outdoor caged study Eulimnadia sp. Takahashi ZR-515 granular/acre 1973

Conchostraca Clam shrimp, Miura & technical X irrigated pasture study Eulimnadia sp. Takahashi ZR-515 1973

Copepoda Copepods, Miura & technical X irrigated pasture study Cyclops sp. Takahashi ZR-515 1973

Copepoda Copepods, Miura & technical 0.1 lb EC/acre X pond study Cyclops sp. Takahashi ZR-515 1973

Copepoda Copepods, Miura & technical 5 laboratory acute toxicity tests: LC50 Cyclops sp. Takahashi ZR-515 concentrations = 4.60 ppm; test duration 24 hours 1973 Copepoda Copepods, Miura & technical 0.1 ppm X outdoor test in artificial container; no Cyclops sp. Takahashi ZR-515, detectable effects 1973 10% flowable liquid (slow release)

Decapoda Crayfish Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Procambarius 1977 population increases attributed to clarki and reduced predator

Cambarellus sp.) populations Decapoda Fiddler Crab McAlonan et Altosid SR- 0.024 to 0.384 X no significant mortality nor frequency at. 1976 10 lbs AI/A; 3 of ecdysis affected treatments at 2- week intervals

Decapoda Fiddler Crab McAlonan et Altosid 10- 0.012 to 0.120 X caused no mortality at. 1976 F lbs AI/A

Decapoda Grass Shrimp McKenney & methoprene, 1000  g/l X lab study; all larvae died (Palaemonetes Matthews technical pugio) 1990 grade

Decapoda Clam Shrimp Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Palaemonetes 1977 paludosus)

Decapoda Grass Shrimp McKenney & methoprene, 0.1  g/l lab study; 100 g/l rate had (Palaemonetes Matthews technical 10 g/l significant effect; other rates had no pugio) 1990 grade 100  g/l effect

Decapoda Grass Shrimp McKenney & A.L.L. 1000  g/l X lab study; all larvae died (Palaemonetes Matthews pugio) 1990

Decapoda Grass Shrimp McAlonan et Altosid SR- 0.024 to 0.384 X no significant mortality nor frequency at. 1976 10 lbs AI/A; one of ecdysis affected series of 4

treatments & second series of 3 treatments at 2-week intervals

Decapoda Grass Shrimp McKenney A.L.L. 8, 16, 32, 62, X lab study; significant mortality was (Palaemonetes & Celestial 125, 250  g/l seen after 2 days exposure for 250, pugio) 1993 after 4 days for 62 & greater, and after 8 days for all conc. ; both dry weights & daily growth rates for 1- and 9-day old larvae significantly reduced by 8 g/l and greater conc. exposures

Decapoda Grass Shrimp McKenney & A.L.L. 0.1  g/l X lab study; no effect (Palaemonetes Matthews 10 g/l pugio) 1990 100  g/l

Decapoda Grass Shrimp McAlonan et Altosid 10- 0.048 to 0.120 X produced greater than 60% mortality at. 1976 F lbs AI/A

Decapoda Mud Crab Celestial & A.L.L. varying conc.: X lab study; no statistically significant (Rhithropanopeus McKenney 0.1, 1.0, 10.0 reductions in survival rates; although harrisii) 1994 g/l zoeal stages I & II showed reduced survival rates; no significant differences in cumulative development duration at these conc.

Decapoda Mud Crab Celestial & A.L.L. 100  g/l X lab study; significant reductions in (Rhithropanopeus McKenney survival for all development stages harrisii) 1994 except zoeal stage II; significant development duration, increased total development duration by 4 days

Decapoda Mud Crab Celestial & A.L.L. 1000  g/l X lab study; no larvae survived beyond (Rhithropanopeus McKenney zoeal stage I harrisii) 1994 Decapoda Mysidiopsis bahia McKenney A.L.L. varying: X lab study; no significant effects on & Celestial 2,4,8,16,32, 62 mortality through life cycle 1996 g/l

Decapoda Mysidiopsis bahia McKenney A.L.L. varying: X lab study; reproduction affected by & Celestial 2,4,8,16,32, 62 sublethal concentrations greater than 1996 g/l 2 g/l; average time to first brood release significantly delayed for all conc. except 2 & 16 g/l; brood size reduced in all conc. greater than 8 g/l.

Decapoda Mysidiopsis bahia McKenney A.L.L. 125  g/l X lab study; 100% mortality by 4 days & Celestial of exposure 1996

Decapoda Mysidiopsis bahia McKenney A.L.L. varying: lab study; 62 g/l significantly & Celestial 2,4,8,16,32, 62 affected dry weights after 15 days of exposure; other concentrations

1996 g/l had no effect Mysidacea Taphromysis Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; louisiana (opossum 1977 greater numbers collected in open shrimp) water habitats, but slightly higher mortality occurred in emergent plant habitat

Notostraca Tadpole shrimp, Miura & technical 5 laboratory acute toxicity tests: Triops Takahashi ZR-515 concentrations LC50 = 5.00 ppm; test duration longicaudatus 1973 24-96 hours

Notostraca Tadpole shrimp, Miura & technical 0.1 lb EC/acre X outdoor caged study Triops Takahashi ZR-515 longicaudatus 1973

Notostraca Tadpole shrimp, Miura & technical X irrigated pasture study Triops Takahashi ZR-515 longicaudatus 1973

Ostracoda Ostracod Norland & Altosid EC 0.1 ppm X repeated treatments of experimental (Cyprinotus sp.) Mulla 1975 ponds; (information from abstract)

Ostracoda Seed shrimp, Miura & technical X irrigated pasture study Cypricercus sp. Takahashi ZR-515 1973

Ostracoda Seed shrimp, Miura & technical 5 laboratory acute toxicity tests: LC50 Cypricercus sp. Takahashi ZR-515 concentrations = 1.50 ppm; test duration 24

1973 hours Ostracoda Seed shrimp, Miura & technical 0.7 lb corncob X outdoor caged study Cypricercus sp. Takahashi ZR-515 granular/acre 1973

Berosus sp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; Insects 1977 Coleoptera Coleoptera Berosus exiguus Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Coleoptera Berosus infuscatus Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Coleoptera Coleoptera Hershey et Altosid - 3 5.820.44 X natural wetlands; aerial application, 6 al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study); 46% reduction in 1992; 59% reduction in 1993; 48% reduction over 4-yr period

Coleoptera Copelatus sp. Miura & technical X irrigated pasture study Takahashi ZR-515 1973

Coleoptera Dytiscid beetle Norland & Altosid EC 0.1 ppm X repeated treatments of experimental (Laccophilus sp.) Mulla 1975 ponds; eliminated from treated ponds (information from abstract)

Coleoptera Enochrus Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; blatchleyi 1977 Coleoptera Hydrocanthus spp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Coleoptera Hydrovatus Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; cuspidatus 1977

Coleoptera Laccophilus Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; proximus 1977

Coleoptera Liodessus affinis Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977 population increases attributed to reduced predator populations

Coleoptera Lissorhoptrus spp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977 Coleoptera Lixellus sp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Coleoptera Noteridae Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Coleoptera Predaceous water Miura & technical X irrigated pasture study beetle, Laccophilus Takahashi ZR-515 sp. 1973

Coleoptera Predaceous water Miura & technical 5 laboratory acute toxicity tests:

beetle, Takahashi ZR-515 concentrations LC50 = 2.00 ppm; test duration Laccophilus sp. 1973 48-72 hours Coleoptera Scavenger beetle Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Tropisternus 1977 lateralis)

Coleoptera Suphisellus spp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977 Coleoptera Water scavenger Miura & technical 5 laboratory toxicity tests: 0% mortality beetle, Takahashi ZR-515 concentrations at 1 ppm; test duration 120 hours Tropisternus 1973 lateralis

Coleoptera Water scavenger Miura & technical 5 laboratory toxicity tests: 57% beetle, Helophorus Takahashi ZR-515 concentrations mortality at 0.8 ppm; 48% at 2.5 ppm; sp. 1973 test duration 72-96 hours

Coleoptera Water scavenger Miura & Miura & X irrigated pasture study beetle, Helophorus Takahashi Takahashi sp. 1973 1973

Coleoptera Water scavenger Miura & technical 5 laboratory toxicity tests: 0% beetle, Hydrophilus Takahashi ZR-515 concentrations mortality at 24 ppm; 100% at 100 triangularis 1973 ppm ; test duration 144-240 hours

Coleoptera Water scavenger Miura & technical X irrigated pasture study beetle, Takahashi ZR-515

Tropisternus 1973 lateralis Coleoptera Water scavenger Miura & technical X irrigated pasture study beetle, Hydrophilus Takahashi ZR-515 triangularis 1973

Coleoptera Whirligig beetle, Miura & technical 5 laboratory toxicity tests: 100% Gyrinus punctellus Takahashi ZR-515 concentrations mortality at 6 ppm; test duration 48 1973 hours

Diptera Anopheles sp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Diptera Biting Midges - Hershey et Altosid - 3 5.820.44 X natural wetlands; aerial application, Ceratopogonids al. 1998 wk release kg/ha 6 treatments per year for 3 years granules (long-term effects study); reduced in 1992 & 1993; 3-yr period showed reduction of 55%

Diptera Chironomid Norland & Altosid EC 0.1 ppm X repeated treatments of Mulla 1975 experimental ponds; twofold reduction by treatment (information from abstract)

Diptera Chironomidae Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, 6 al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects

study); seasonal significant reduction in 1992 & 1993, and for overall 3-yr treatment period Diptera Chironomids Ali 1991 XR 0.82 kg AI/ha X experimental pond; 38-96% Briquets control for 7 weeks

Diptera Chironomids Ali 1991 Pellets 0.22 kg AI/ha X experimental pond; 64-98% control for 7 weeks Diptera Chironomids Ali 1991 Granular 0.17 kg AI/ha X experimental pond; lost (SAN 810 I rd 1.3 GR) effectiveness in 3 week post- treatment Diptera Chironomids Ali 1991 A.L.L. 0.28 kg AI/ha X experimental pond; returned to pre-treatment levels in 3rd week after treatment Diptera Chironomids Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Diptera Chironomids Ali 1991 A.L.L. 0.015 kg AI/ha X experimental pond

Diptera Crane flies -- Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, Tipulidae al. 1998 wk release kg/ha 6 treatments per year for 3 years granules (long-term effects study); reduced in 1992 & 1993; 3-yr period showed reduction of 73%

Breaud et al. 6 aerial applications over 18

Diptera Culex salinarius 1977 methoprene 28 gm AI/ha X months; Diptera Diptera Hershey et Altosid - 3 5.820.44 X natural wetlands; aerial application, 6 al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study); exhibited 3-yr reduction of 66%

Diptera Flower fly, Nylota Miura & technical 5 laboratory toxicity tests: 0% sp. Takahashi ZR-515 concentrations mortality at 6 ppm; test duration 72 1973 hours

Diptera Flower fly, Nylota Miura & technical X irrigated pasture study sp. Takahashi ZR-515 1973

Diptera Green heads - Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; Dolichopodidae 1977

Diptera Lispe sp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Diptera Midge, Miura & technical irrigated pasture study; some dead Chironomus Takahashi ZR-515 pupae stigmaterus 1973

Diptera Midge, Miura & technical 5 laboratory toxicity tests: 50% Chironomus Takahashi ZR-515 concentrations mortality at 0.01 ppm; test duration stigmaterus 1973 288 hours

Mothfly, Pericoma Miura & technical 5 laboratory toxicity tests: 50%

Diptera sp. Takahashi ZR-515 concentrations mortality at 0.1 ppm; test duration 480 1973 hours Diptera Nematocera Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, 6 al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study); exhibited 3-yr reduction of 68%

Diptera Notophila ap. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Diptera Predatory Hershey et Altosid - 3 5.820.44 X natural wetlands; aerial application, 6 chironomids al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study); seasonal significant reduction in 1992 & 1993, and for overall 3-yr treatment period

Diptera Sandflies Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Psychoda sp.) 1977 population increases attributed to reduced predator populations

Diptera Shorefly, Miura & technical irrigated pasture study; some dead Brachydeutera Takahashi ZR-515 pupae argentata 1973

Diptera Shorefly, Miura & technical 5 laboratory toxicity tests: 70% Brachydeutera Takahashi ZR-515 concentrations mortality at 0.01 ppm; test

argentata 1973 duration 504 hours Diptera Soldier flies Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Eulalia sp.) 1977

Diptera Soldier flies -- Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, Stratiomyidae al. 1998 wk release kg/ha 6 treatments per year for 3 years granules (long-term effects study); no effect seen until 1993, then showed 71%, with overall 3- yr reduction of 44%

Ephemeroptera Mayflies -- Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, Brachycera al. 1998 wk release kg/ha 6 treatments per year for 3 years granules (long-term effects study); no effect seen until 1993, then showed 69%, with overall 3- yr reduction of 36%

Ephemeroptera Mayflies Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Callibaetis sp.) 1977

Ephemeroptera Mayfly nymphs, Miura & technical 5 laboratory toxicity tests: 0% at 10 Callibaetis sp. Takahashi ZR-515 concentrations ppm; test duration 48 hours 1973

Ephemeroptera Mayfly (Caenis Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; sp.) 1977

Ephemeroptera Mayfly nymphs, Miura & technical X irrigated pasture study

Callibaetis sp. Takahashi ZR-515 1973 Ephemeroptera Mayfly Norland & Altosid EC 0.1 ppm X repeated treatments of experimental (Callibaetis Mulla 1975 ponds; mortality in early and late pacificus) instars during winter; effect lessened with rising water temperatures (information from abstract)

Hemiptera Backswimmer, Miura & technical technical ZR- X outdoor test in artificial container; no Notonecta unifasciata Takahashi ZR-515, 515, 10% visible effects on populations 1973 10% flowable liquid flowable (slow release) liquid (slow release)

Hemiptera Backswimmer, Miura & technical 5 laboratory acute toxicity tests: LC50 Notonecta unifasciata Takahashi ZR-515 concentrations = 1.20 ppm; test duration 24 hours 1973 Hemiptera Backswimmer, Miura & technical X irrigated pasture study Notonecta unifasciata Takahashi ZR-515 1973

Hemiptera Buenoa spp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Hemiptera Corixids Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 (Trichocorixa 1977 months; population increases

louisianae) attributed to reduced predator populations Hemiptera Giant water bug Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Belostoma 1977 testaceum)

Hemiptera Water treader Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Mesovelia mulsanti) 1977

Hemiptera Waterboatman, Miura & technical 0.1 ppm X outdoor test in artificial containers; Corisella decolor Takahashi ZR-515, no visible effects on populations 1973 10% flowable liquid (slow release)

Hemiptera Waterboatman, Miura & technical X irrigated pasture study Corisella decolor Takahashi ZR-515 1973

Hemiptera Waterboatman, Miura & technical 5 laboratory acute toxicity tests: Corisella decolor Takahashi ZR-515 concentrations LC50 = 1.65 ppm; test duration 1973 24-96 hours

Odonata Coenagrionidae Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Odonata Damselfly Miura & technical 5 laboratory toxicity tests: 0%

nymphs, Argia sp. Takahashi ZR-515 concentrations mortality at 1 ppm; test duration 48 1973 hours Odonata Dragonflies Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; (Belonia & Anax) 1977

Odonata Dragonfly nymphs, Miura & technical 5 laboratory toxicity tests: 0% mortality Orthemis sp. Takahashi ZR-515 concentrations at 24 ppm; 30 % at 100 ppm; test 1973 duration 72 hours

Odonata Odonata naiads Norland & Altosid EC 0.1 ppm X repeated treatments of experimental Mulla 1975 ponds; (information from abstract)

Odonata Pachydiplax sp. Breaud et al. methoprene 28 gm AI/ha X 6 aerial applications over 18 months; 1977

Miscellaneous Non-dipteran Hershey et Altosid - 3 5.820.44 X natural wetlands; aerial application, 6 predators al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study); significant reduction in 1992 (46%) and 1993 (64%),

Miscellaneous Total predatory Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, 6 insects al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study); significant reduction in 1992 (65%) and 1993 (77%), and over 3-yr period (62%)

Miscellaneous Non-insect macro- Hershey et Altosid 3- 5.820.44 X natural wetlands; aerial application, 6 invertebrates al. 1998 wk release kg/ha treatments per year for 3 years (long- granules term effects study);

Aquatic Miura & technical 5 laboratory toxicity tests: 0% Annelids earthworms, Takahashi ZR-515 concentrations mortality at 100 ppm; test duration Oligochaeta Aulophorus sp. (3 1973 168 hours species)

Oligochaeta Mud worm, Miura & technical 5 laboratory toxicity tests: 0% mortality Tubifex tubifex Takahashi ZR-515 concentrations at 10 ppm; test duration 168 hours 1973

Rhynochobdellida Leeches, Miura & technical 5 laboratory toxicity tests: 0% Helobdella Takahashi ZR-515 concentrations mortality at 1 ppm; test duration 72 stagnalis 1973 hours

Nematodes Miura & technical X irrigated pasture study Aschelminths Takahashi ZR-515 Nematoda 1973

Rotifera Rotifer, Philodina Miura & technical 5 laboratory toxicity tests: 5% sp. Takahashi ZR-515 concentrations mortality at 100 ppm; test duration 1973 48-72 hours

Brown planarian, Miura & technical 5 laboratory toxicity tests: 33% Flatworms Dugesia tigrina Takahashi ZR-515 concentrations mortality at 10 ppm; test duration 168 Tricladida 1973 hours

Protozoa Paramecia, Miura & technical 5 laboratory acute toxicity tests: LC50 Hymenostomatida Paramecium sp. Takahashi ZR-515 concentrations = 1.25 ppm; test duration 48 hours 1973 Diatom, Diatoma Miura & technical 0.1 ppm X lab study; no visible effects after 1 Phytoplankton vulgare Takahashi ZR-515 solution week 1973

Phytoplankton Green algae (3 Miura & technical 0.1 ppm X lab study; no visible effects after 1 species), Takahashi ZR-515 solution week Pithaphora 1973 ocdogonia, Spirogyra sp., Hydrodictyon reticulatum

Temephos

Temephos is an organophosphate pesticide, which functions by competing with acetylcholine for cholinesterase, the enzyme that transmits nerve impulses across synapses to other nerves and muscles (known as a Acholinesterase inhibitor@). While acetylcholine is present, the neurons continue to be stimulated; paralysis results from the failure of cholinesterase to destroy the acetylcholine (Dale and Hulsman 1990). Temephos is a general use pesticide; temephos-containing products are moderately toxic and are labeled with WARNING, due to the high toxicity of xylene, one of the carrier compounds found in many trade products. Toxicological effects include both acute and chronic toxicity (Pesticide Information Profiles, EXTOXNET).

Synopsis of Non-target Effects

Effects of temephos on some non-target organisms are presented in Table 4. Moderate toxicity to birds and fish; was shown to accumulate in tissues of fish and snails, but effect was reversible. Wide range of crustaceans, insects and mollusks were affected by temephos. Some crustacean and mollusks exhibited sub-lethal effects (slowed responses resulting in increased susceptibility to predation).

Label Application Rate

5% Skeeter Abate7 (Abate7 5-BG) is used for the control of mosquito and midge larvae. It is toxic to birds and fish; fish and other aquatic organisms in water treated with this product may be killed. Consult state fish and game agency before applying this product to waters or wetlands. Do not use on crops used for food, forage or pasture.

Habitat: standing water, shallow ponds, lakes and woodland pools. Application Rate: 2 lbs/acre

Habitat: tidal waters, marshes, swamps and waters high in organic content. Application Rate: 4 lbs/acre

Habitat: highly-polluted water. Application Rate: 10 lbs/acre.

1% Skeeter Abate7 (Abate7 1-BG) is used for the control of mosquito and midge larvae. It is toxic to birds and fish; fish and other aquatic organisms in water treated with this product may be killed. Consult state fish and game agency before applying this product to waters or wetlands. Do not use on crops used for food, forage or pasture.

Habitat: standing water, shallow ponds, lakes, woodland pools, catch basins. Application Rate: 5 - 10 lbs/acre

Habitat: tidal waters, marshes, swamps and waters high in organic content. Application Rate: 10 - 20 lbs/acre

Web Site:

Pesticide Information Profiles, EXTOXNET http://ace.ace.orst.edu/info/extoxnet/pips/ghindex.html

References Cited

Ali, A. and M.S. Mulla. 1978. Effects of chironomid larvicides and diflubenzuron on nontarget invertebrates in residential-recreational lakes. Environ. Entomol. 71:21- 27.

Ali, A. and M.S. Mulla. 1977. Chemical control of nuisance midges in the Santa Ana River Basin, Southern California. J. Econ. Entomol. 70:191-195.

Balcomb, R., R. Stevens and C. Bowen. 1984. Toxicity of 16 granular insecticides to wild- caught songbirds. Bull. Environ. Contam. Toxicol. 33:302-307.

Campbell, B.C. and R.F. Denno. 1976. The effect of Temephos and Chlorpyrifos on the aquatic insect community of a New Jersey salt marsh. Environ. Entomol. 5:477- 483

Chambers, H. And D.L. Fabacher. 1972. Midge larvae control in commercial catfish ponds: toxicity of Abate7 to channel catfish (Ictularus punctatus). Proc. Annu. Conf. Southeast. Assoc. Game Fish Comm. 26:399-401. (Abstract)

Dale, P.E.R. and K. Hulsman. 1990. A critical review of salt marsh management methods for mosquito control. Review in Aquatic Sciences 3:281-311.

Dale, W.E., J.W. Miles and G.O. Guerrant. 1974. Monitoring of residues of Abate in streams treated for Similium control. IN Environmental Quality and Safety Supplement, Vol. III, Pesticides. Pp 780-783.

Denno, R.F. 1974. Initial studies of Abate in a salt-marsh ecosystem: sampling the nontarget component. Proc. N.J. Mosq. Exterm. Assn. 61:138-144.

Fales, J.H., P.J. Spangler, O.F. Bodenstein, G.D. Mills, Jr. and C.G. Durbin, Jr. 1968. Laboratory and field evaluation of Abate against a backswimmer Notonecta undulata Say (Hemiptera: Notonectidae). Mosq. News 28:77-81.

Fleming, W.J., G.H. Heinz, J.C. Franson, and B.A. Rattner. 1985. Toxicity of Abate 4E (temephos) in mallard ducklings and the influence of cold. Environ. Toxicol. Chem. 4:193-199.

Frank, A.M. and R.D. Sjogren. 1978. Effect of temephos and chlorpyrifos on Crustacea. Mosq. News 38:138-139.

Franson, J.C., J.W. Spann, G.H. Heinz, C. Bunck and T. Lamont. 1983. Effects of dietary AbateR on reproductive success, duckling survival, behavior, and clinical pathology in game-farm mallards. Arch. Environ. Contam. Toxicol. 12:529-534.

Fitzpatrick, G. and D.J. Sutherland. 1976. Uptake of the mosquito larvicide temefos by the salt marsh snail, New Jersey--1973-74. Pesticide Monitoring J. 10:4-6.

Hanazato, T., T. Iwakuma, M. Yasuno, and Mitsuru Sakamoto. 1989. Effects of temephos on zooplankton communities in enclosures in a shallow eutrophic lake. Environ. Pollut. 59:305-314.

Hill, E.F. 1971. Toxicity of selected mosquito larvicides to some common avian species. J. Wildl. Manage. 35:757-762.

Kpekata, A.E. 1983. Acute toxicity of O,O=-(thio-di-4,1-phenylene) bis (O,O-dimethyl phosphorothioate) (temephos) to Lebistes reticulatus and Sarotherodon galilea. Bull. Environ. Contam. Toxicol. 31:120-124.

Levy, R. And T.W. Miller, Jr. 1977. Susceptibility of the mosquito nematode, Romanomermis culicivorax (Mermithidae) to pesticides and growth regulators. Environ. Entomol. 6:447- 448.

Mathavan, S. And E. Jayakumar. 1987. Long-term effects of pesticides (fenthion and temephos) on growth and fecundity of an aquatic bug Laccotrephes griseus (Guerin). Ind. J. Exp. Biol. 25:48-51.

Mohsen, A.H. and M.S. Mulla. 1981. Toxicity of blackfly larvicidal formulations to some aquatic insects in the laboratory. Bull. Environ. Contam. Toxicol. 26:696- 703.

Muirhead-Thomson, R.C. 1979. Experimental studies on macroinvertebrate predator- prey impact of pesticides. The reactions of Rhyacophilia and Hydropsyche (Trichoptera) larvae to Simulium larvicides. Can. J. Zoo. 57:2264-2270.

Nelson, F.R.S., J. Gray and F. Aikhionbare. 1994. Tolerance of the planarian Dugesia tigrina (Tricladida: Turbellaria) to pesticides and insect growth regulators in a small-scale field study. J. Am. Mosq. Control Assoc. 10:104-105.

Pierce, R.H., R.C. Brown, K.R. Hardman, M.S. Henry, C.L.P. Palmer, T.W. Miller, and G. Wichterman. 1989. Fate and toxicity of temephos applied to an intertidal mangrove swamp. J. Amer. Mosq. Control Assn. 5:569-578.

Sanders, H.O., D.F. Walsh, and R.S. Campbell. 1981. Abate: Effects of the organophosphate insecticide on bluegills and invertebrates in ponds. U.S. Fish & Wildl. Serv. Tech. Pap. 104, 6pp.

Tietze, N.S., P.G. Hester, C.F. Hallmon, M.A. Olson, and K.R. Shaffer. 1991. Acute toxicity of mosquitocidal compounds to mosquitofish, Gambusia affinis. J. Amer. Mosq. Control Assn. 7:290-293.

Von Windeguth, D.L. and R.S. Patterson. 1966. The effects of two organic phosphate insecticides on segments of the aquatic biota. Mosq. News 26:377-380.

Wall, W.J., Jr. and V.M. Marganian. 1973. Control of salt marsh Culicoides and Tabanus larvae in small plots with granular organophosphorus pesticides, and the direct effect on other fauna. Mosq. News 33:88-93.

Wall, W.J. Jr. and V.M. Marganian. 1971. Control of Culicoides mellus (Coq.) (Diptera: Ceratopogonidae) with granular organophosphorus and the direct effect on other fauna. Mosq. News 31:209-214.

Wallace, R.R., A.S. West, A.E.R. Doune and H.B.N. Hynes. 1973. The effects of experimental blackfly (Diptera: Simuliidae) larviciding with Abate, Dursban, and methoxychlor on stream invertebrates. Can. Entomol. 105:817-831.

Ward, D.V. and D.A. Busch. 1976. Effects of temefos, an organophosphorous insecticide, on survival and escape behaviors of the marsh fiddler crab Uca pugnax. Oikos 27:331- 335.

Ward, D.V. and B.H. Howes. 1974. The effects of Abate, an organophosphorous insecticide, on marsh fiddler crab populations. Bull. Environ. Contam. Toxicol. 12:694-697.

Ward, D.V., B.L. Howes, and D.F. Ludwig. 1976. Interactive effects of predation pressure and insecticide (temefos) toxicity on populations of the marsh fiddler crab Uca pugnax. Marine Biol. 35:119-126.

Yasuno, M., Y. Sugaya and T. Iwakuma. 1985. Effects of insecticides on benthic community in a model stream. Environ. Pollut. 38:31-43.

Table 4. Non-target Effects of Temephos

Classification Organism Reference Formulation Application Adverse No Comments Rate (study) Effects Effects

Blue Jays Hill 1971 technical 5 conc. X 30 ppm killed all birds in test; birds fed for Birds grade tested 5 days on toxic diet Aves Aves Bobwhites Hill 1971 technical 5 conc. LC = 1,540 ppm; birds fed for 5 days on grade tested 50 toxic diet Aves Cardinals Hill 1971 technical 5 conc. LC = 76 ppm; birds fed for 5 days on grade tested 50 toxic diet Aves House Sparrows Hill 1971 technical 5 conc. LC = 47 ppm; birds fed for 5 days on grade tested 50 toxic diet

Aves House Sparrows Balcomb et al granules 0.078 mg X no mortality in doses up to 40 granules 1984 4%AI mean granule weight Aves Mallard Fleming, et Abate 4E 0.1ppm; 1 treatments of 10 ppm or less did not enhance ducklings al. 1985 ppm; 10 cold effects on ducklings nor depressed brain ppm; cholinesterase (ChE); 100pm did significantly 100ppm affect cold tolerance and depressed brain ChE and degree of inhibition was less than previously used to document death from

anticholinesterase insecticides Aves Mallard adults Franson et Abate 4E 1 ppm & 10 females took longer to complete egg- laying al. 1983 ppm with 10 ppm concentration diet

Aves Mallard Franson et Abate 4E 1 ppm & 10 ducklings in both treatment diets had 20% ducklings al. 1983 ppm body weight (not statistically significant but noteworthy); survivability reduced 40% in both treatments

Aves Red-winged Balcomb et al granules 0.078 mg X no mortality in doses up to 40 granules Blackbirds 1984 4%AI mean granule weight

Natrix sipedon Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Reptiles 1968 (EC post-application; no dead found Squamata Testudines Chrysemys picta Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. 1968 (EC post-application; no dead found

Rana clamitans Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Amphibians 1968 (EC post-application; no dead found Anura Caudata Triturus Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. viridescens 1968 (EC) post-application; no dead found

Fish Guppies Kpekata temephos acute effect lab study; 96-hour LC50 = Atheriniformes (Sarotherodon 1983 0.47 mg/l (information from abstract) galilaea)

Atheriniformes Guppies Kpekata temephos acute effect lab study; 96-hour LC50 = (Lebistes 1983 1.9 mg/l (information from abstract) reticulatus)

Atheriniformes Guppy (Lebistes Von Abate, 0.20-0.25 X safe at 0.1 ppm; 24-hour LD50 = 200 ppm reticulatus) Windeguth technical lb/acre + and material (conc. of Patterson 0.1 ppm in 1966 1 ft water depth or 0.01 ppm in 10 ft. depth)

Atheriniformes Juvenile snook Pierce et temephos, X no mortality observed (information from (Centropomis al. 1989 aerially abstract) undecimalis) applied

Atheriniformes Killifish Wall and Abate 1% 0.3 lb/acre fish in 2 traps were dead, but those in 3rd trap (Fundulus spp.) Marganian on sand survived with no apparent effect for 7 days; 1973 granules unable to attribute mortality to pesticide

Atheriniformes Killifish Wall and Abate 1% 0.4 lb/ac X (Fundulus spp.) Marganian on sand 1971 granules

Atheriniformes Mosquito fish Von Abate, 0.20-0.25 X safe at 0.1 ppm; 24-hour LD50 = 200 ppm (Gambusia Windeguth technical lb/acre + affinis) and material (conc. of Patterson 0.1 ppm in

1966 1 ft water depth or 0.01 ppm in 10 ft. depth) Atheriniformes Mosquitofish Tietze et al Abate 4-E various X no affect at recommended application rate; 1991 24-hour LC50 = 5.60 ppm

Atheriniformes Mummichog Lee & Scott technical 96-hour LC = 0.04 mg/L; no effect (Fundulus 1989 grade 50 heteroclitus) concentration = 0.02 mg/L

Atheriniformes Sheepshead Pierce et temephos, X no mortality observed (information from minnow al. 1989 aerially abstract) (Cyprinodon applied variegatus)

Cypriniformes Catfish Chambers Abate R LC50 determined to be 5-7ppm in & laboratory Fabacher 1972

Perciformes Blue gill Von Abate, 0.20-0.25 X safe at 0.1 ppm; 24-hour LD50 = 200 ppm (Lepomis Windeguth technical lb/acre + macrochirus) and material (conc. of Patterson 0.1 ppm in 1966 1 ft water depth or 0.01 ppm in

10 ft. depth) Perciformes Bluegills Sanders et Abate EC 18 g/ha X 3 treatments in experimental ponds; initially al 1981 more rapid growth and higher reproduction presumably from increased food (dead Dipterans), but declined after 3rd treatment attributed to decline in Dipterans

Perciformes Bluegills Sanders et Abate EC 180 g/ha 3 treatments in experimental ponds; brain al 1981 acetylcholinesterase activity depressed 40% when water temperature exceeded 20oC; lower growth and production rates attributed to greater losses of Dipterans from first treatment

Perciformes Largemouth bass Von Abate, 0.20-0.25 X safe at 0.1 ppm; 24-hour LD50 = 200 ppm (Micropterus Windeguth technical lb/acre + salmoides) and material (conc. of Patterson 0.1 ppm in 1966 1 ft water depth or 0.01 ppm in 10 ft. depth)

Water Mites -- Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Arachnids Hydrachnidae 1968 (EC) post-application; 1 species, no dead found Acarina

Ribbed mussel Wall and Abate 1% Mollusks

Anisomyaria (Modiolus Marganian on sand 0.3 lb/acre X demissus) 1973 granules Basommatophora Snails -- Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Planorbidae 1968 (EC) post-application; 1 species, no live individuals in samples

Basommatophora Snails -- Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Physidae 1968 (EC) post-application; 1 species, no live individuals in samples Gastropoda Snail (Melampus Fitzpatrick Abate 2% 0.10 lb/acre; X uptake detectable 1 day after 1st treatment; bidentatus) and granular 10 residues persisted for more than 5 weeks Sutherland applications after last treatment; 1976 at 2 week intervals

Gastropoda Snail (Melampus Fitzpatrick Abate 0.032 X uptake detectable 6 days after 2nd treatment; bidentatus) and emulsion lb/acre; 4 residues rose gradually as number of Sutherland applications treatments increased, then decreased below 1976 at 2 week detection limit 3-weeks after last treatment; intervals data indicate significant but reversible decline in population density

Mesogastropoda Mud snail Wall and Abate 1% 0.3 lb/acre X (Nassarius Marganian on sand obsoletus) 1973 granules

Mesogastropoda Mud snail Wall and Abate 1% 0.4 lb/ac those confined in traps were alive but

(Nassarius Marganian on sand some appeared to have slowed responses obsoletus) 1971 granules Mesogastropoda Periwinkle (snail) Wall and Abate 1% 0.3 lb/acre X (Littorina littorea) Marganian on sand 1973 granules

Crustaceans Sideswimmer Ali and temephos 0.28 kg X tolerant to temephos; higher concentration Amphipoda (Hyallela Mulla AI/ha used in lake fingers, lower concentration in azteca) 1978 (0.0092 main lake area; (information from abstract) ppm) & 0.17 kg AI/ha (0.0042 ppm)

Amphipoda Sideswimmer Von Abate, 0.20-0.25 safe at 0.1 ppm; 24-hour LD50 = 0.65 (Hyallela Windeguth technical lb/acre ppm; LD90 = 2-2.5 ppm azteca) and material (conc. of Patterson 0.1 ppm in 1966 1 ft water depth or 0.01 ppm in 10 ft. depth)

Calanoida Calanoid Hanazato Abate 500 g AI/l X shallow lake; eliminated; nauplii showed et al. 1989 slight recovery by end of experiment

Calanoida Diaptomus spp. Ali and temephos 0.28 kg higher concentration used in lake fingers, Mulla AI/ha lower concentration in main lake area;

1978 (0.0092 (information from abstract) ppm) & 0.17 kg AI/ha (0.0042 ppm)

Cladocera Bosmina Ali and temephos 0.28 kg X higher concentration used in lake fingers, longirostris Mulla AI/ha lower concentration in main lake area; 1978 (0.0092 (information from abstract) ppm) & 0.17 kg AI/ha (0.0042 ppm)

Cladocera Cyclops sp. Ali and temephos 0.28 kg X higher concentration used in lake fingers, Mulla AI/ha lower concentration in main lake area; 1978 (0.0092 (information from abstract) ppm) & 0.17 kg AI/ha (0.0042 ppm)

Cladocera Water fleas -- Hanazato Abate 500 g AI/l X shallow lake; all eliminated; had not Cladocerans et al. 1989 recovered by end of experiment

Cladocera Water flea Ali and temephos 0.28 kg X higher concentration used in lake fingers,

(Daphnia pulex) Mulla AI/ha lower concentration in main lake area; 1978 (0.0092 population reduced in fingers but recovered ppm) & within 1-3 weeks (information from abstract) 0.17 kg AI/ha (0.0042 ppm)

Cladocera Water flea Ali and temephos 0.28 kg X higher concentration used in lake fingers, (Daphnia Mulla AI/ha lower concentration in main lake area; galeata) 1978 (0.0092 population reduced in fingers but recovered ppm) & within 1-3 weeks (information from abstract) 0.17 kg AI/ha (0.0042 ppm)

Crustacea Crustacea Frank and temephos 0.025 lb X copepods, ostracods, amphipods, & Sjogren AI/acre cladocerans; no effect on occurrence (numbers 1978 not studied)

Cyclopoida Cyclopoids Hanazato Abate 500 g AI/l X shallow lake; eliminated; nauplii showed et al. 1989 slight recovery by end of experiment

Cyclopoida Paracyclops Yasuno et temephos 5 mg/l; 30 X model stream study fimbriatus al 1985 min exposure

Decapoda Brown shrimp Pierce et temephos, X no mortality observed (information from (Panaeus aerially

aztecus) al. 1989 applied abstract) Decapoda Fiddler Crab Ward and Abate 99% 12 X 24-hour lab experiments; number of crabs Busch pure concentratio either dead or not responding to stimulus 1976 crystalline ns from 0.5 (EC) increased with increasing temephos powder ppm to 15 concentration; LC20 = 2.06 ppm; LC50 = ppm 9.12 ppm; LC80 = 39.8 ppm; EC20 = 1.10 ppm; EC50 = 4.31 ppm; EC80 = 16.6 ppm

Decapoda Fiddler Crab (Uca Wall and Abate 1% 0.3 lb/acre few dead crabs found in treated area sp.) Marganian on sand 1973 granules

Decapoda Fiddler Crab Wall and Abate 1% 0.4 lb/ac X numerous dead crabs found in treated areas; (Uca pugilator) Marganian on sand however, those confined in traps were not 1971 granules visibly affected at 7 days when released

Decapoda Fiddler Crab Ward et al. Abate 2% 0.1 lb X field experiment; population reduced 14% 1976 granular AI/acre after 2nd application and 30% after 4th application; conclusion that temephos has primarily sublethal effect on crabs that renders them more susceptible to predation

Decapoda Fiddler Crab Ward and Abate 2% 0.1 lb X field test; populations declined over time in Howes granular AI/acre; 3 treated areas 1974 treatments 2 weeks

apart; expected conc. 0.5 ppm Decapoda Freshwater Von Abate, 0.20-0.25 X safe at 0.1 ppm; 24-hour LD = 1.0 ppm; shrimp Windeguth technical lb/acre 50 (Palomonetes and material (conc. of LD90 = 2.0 ppm paludosus) Patterson 0.1 ppm in 1966 1 ft water depth or 0.01 ppm in 10 ft. depth)

Decapoda Grass shrimp Pierce et temephos, X no mortality observed (information from (Palaemonetes al. 1989 aerially abstract) pugio) applied

Mysidacea Mysids Pierce et temephos, X significant mortality at 1 site during 1 of 3 (Mysidopsis bahia) al. 1989 aerially applications monitored (information from applied abstract)

Ostracoda Seed shrimp Ali and temephos 0.28 kg X tolerant to temephos; higher concentration (Cyprinotus sp.) Mulla AI/ha used in lake fingers, lower concentration in 1978 (0.0092 main lake area; (information from abstract) ppm) & 0.17 kg AI/ha (0.0042

ppm) Burrowing Water Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Insects Beetles -- 1968 (EC) post-application;1 species, some mortality Coleoptera Noteridae

Coleoptera Crawling Water Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Beetles -- 1968 (EC) post-application; 5 species found, all of Halipidae which had some mortality

Coleoptera Predaceous Diving Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Beetles -- 1968 (EC) post-application; 13 species found, of Dytiscidae which 6 had some mortality

Coleoptera Water Scavenger Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. post- Beetles -- 1968 (EC) application; 11 species found, all of which Hydrophilidae had some mortality

Coleoptera Whirligig Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Beetles -- 1968 (EC) post-application; 3 species found, of Gyrinidae which 2 had some mortality

Collembola Springtails -- Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Poduridae 1968 (EC) post-application; 1 species, no live individuals found at 24-hour sample

Diptera Black flies Dale et al. Abate 20% 50 ppb for X complete kill 45-50 km downstream 1974 EC 10 min.

Diptera Blackfly Mohsen Abate 50% various X 24- hour LC50 = 0.020 ppm; LC90 = (Simulium and Mulla

argus) 1981 EC dilutions 0.038 ppm

Diptera Blackfly Mohsen Abate 50% various X 24-hour LC50 = 0.0082 ppm; LC90 = (Simulium and Mulla EC dilutions 0.020 ppm virgatum) 1981

Diptera Blackfly Muirhead- Abate 20% various conc. X exposures ranged from 15 minutes to 1 hour; (Simulium spp.) Thompson EC ranging from 24-h mortality ranged from 24% at 1979 0.05 to 0.2 ppm to 98% at 1.0 ppm 2.0 ppm

Diptera Chironomidae Wallace et Abate initial conc. X stream study; al 1973 0.1 ppm

Diptera Chironomids Ali and temephos 0.28 kg X 88-95% control of total midge larvae after 3 Mulla granules 1% AI/surface weeks of treatment; control lasted 5-6 weeks 1977 ha

Diptera Chironomids (3 Yasuno et temephos 5 mg/l; 30 X model stream study species) al 1985 min exposure

Diptera Dipterans Sanders et Abate EC 18 g/ha X 3 treatments in experimental ponds; biomass al 1981 similar to control ponds, however biomass declined rapidly after 3rd treatment

Diptera Dipterans Sanders et Abate EC 180 g/ha X 3 treatments in experimental ponds; al 1981 biomass declined rapidly after 1st

application and remained low Diptera Midge Yasuno et temephos 5 mg/l; 30 X model stream study (Procladius sp.) al 1985 min exposure

Diptera Phantom Midges Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. -- Chaoboridae 1968 (EC) post-application; 1 species, estimated millions dead

Diptera True flies Denno Abate 2% X densities reduced in Spartina patens 1974 on celatom community granules

Ephemeroptera Mayflies Wallace et Abate initial conc. X stream study; al 1973 0.1 ppm

Ephemeroptera Mayfly (Baetis Mohsen Abate 50% various X 24-hour LC50 = 0.0097ppm; LC90 = parvus) and Mulla EC dilutions 0.018 ppm 1981 Hemiptera Laccotrephes Mathavan temephos 0.1 ppm X growth affected; fecundity severely reduced griseus and (information from abstract) Jayakumar 1987

Heteroptera Backswimmers - Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. -Notonectidae 1968 (EC) post-application; 3 species, heavy mortality; in lab experiments 0.02 ppm produced 100% mortality of

backswimmers in 4 days Heteroptera Creeping Water Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Bugs -- Naucoridae 1968 (EC) post-application; 1 species, some mortality

Heteroptera Giant Water Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Bugs B 1968 (EC) post-application; 1 species, some Belostomatidae mortality

Heteroptera Marsh Treaders - Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. -Hydrometridae 1968 (EC) post-application; 1 species

Heteroptera Water Boatmen B Campbell 4E 34.75 g X applications by helicopter Corixidae and Denno emulsifiable AI/ha; 4 (Trichocorixa 1976 conc. biweekly verticalis) treatments

Heteroptera Water Boatmen Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. B Corixidae 1968 (EC) post-application; 4 species, only 1 of which had some live individuals

Heteroptera Water Scorpions Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. -- Nepidae 1968 (EC) post-application;

Heteroptera Water Treaders Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. B Mesovliidae 1968 (EC) post-application; 1 species, no live individuals found post-treatment

Heteroptera Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Water Striders B post-application; 3 species, 2 of which

Gerridae 1968 (EC) had some mortality Homoptera Cicadas/leaf Denno Abate 2% X densities reduced in Spartina patens hoppers 1974 on celatom community granules

Hymenoptera Ants/bees/wasp Denno Abate 2% X densities reduced in both Spartina 1974 on celatom alterniflora and Spartina patens granules communities

Odonata Damselfly -- Fales et al. Abate 4E 0.39 lb/acre X lake application, sampled 24- & 48-hr. Coenagrionidae 1968 (EC) post-application; 2 species, only dead found in samples

Odonata Dragonfly -- Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Aeschnidae 1968 (EC) post-application; 1 species, some mortality

Odonata Dragonfly -- Fales et al. Abate 4E 0.39 lb/acre lake application, sampled 24- & 48-hr. Libellulidae 1968 (EC) post-application; 3 species, 1 only dead found, 1 no dead found, 1 some dead found

Plecoptera Stone flies Wallace et Abate initial conc. X stream study; al 1973 0.1 ppm

Trichoptera Caddis flies Wallace et Abate initial conc. X stream study; al 1973 0.1 ppm

Trichoptera Caddisfly Muirhead- Abate 20% various X exposed for 1 hour to Abate solution; 24- h (Hydropsyche Thompson conc. mortality 29% at 0.2 ppm; 76% at 0.5

pellucidula) 1979 EC ranging from ppm; 74% at 1.0 ppm 0.05 to 1.0 ppm Trichoptera Caddisfly Muirhead- Abate 20% various exposures ranged from 15 minutes to 30 (Rhyacophila Thompson EC conc. minutes; 24-h mortalities reported were dorsalis) 1979 ranging 12% at 0.5 ppm (15 min); 18% at 1.0 from 0.2 ppm (15 min); 8% at 2.0 ppm (15 min); ppm to 2.0 48% at 1.0 ppm (30 min); 33% at 2.0 ppm ppm (30 min)

Trichoptera Caddisfly Mohsen Abate 50% various 24-hour LC50 = 1.3 ppm; LC90 = 4.0 ppm (Hydropsyche and Mulla EC dilutions californica) 1981

Oligochaetes Ali and temephos 0.28 kg X higher concentration used in lake fingers, Annelids Mulla AI/ha lower concentration in main lake area; Oligochaeta 1978 (0.0092 (information from abstract) ppm) & 0.17 kg AI/ha (0.0042 ppm)

Nematode Levy and Abate 0.001 ppm X information from abstract Aschelminths (Romanomermis Miller 1977 Nematoda culicivorax)

Rotifera Rotifers Hanazato Abate 500 g AI/l X shallow lake; original species eliminated

et al. 1989 and replaced by other rotifer species Brown planaria Nelson et temephos 4E X only minimal effect under field conditions Flatworms (Dugesia tigrina) al. 1994 (information from abstract) Tricladida

Microscopic Von Abate, 0.20-0.25 safe at 0.1 ppm; 48-hour LD =50 ppm Plankton 100 plankton (Rotifers, Windeguth technical lb/acre Euglena, Coleps, and material (conc. of Ileonema, etc.) Patterson 0.1 ppm in 1966 1 ft water depth or 0.01 ppm in 10 ft. depth)

Mosquito Bibliography

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*Ali, A. 1991. Activity of new formulations of methoprene against midges (Diptera: Chironomidae) in experimental ponds. J. Amer. Mosq. Control Assn. 7:616-620.

*Ali, A. 1991. Perspectives on Management of Pestiferous Chironomidae (Diptera), An Emerging Global Problem. J. Amer. Mosq. Control Assn. 7:260-281.

@Ali, A. 1981. Bacillus thuringiensis serovar israelensis (ABG-6108) against chironomids and some nontarget aquatic invertebrates. J. Invertebr. Pathol. 38:264-272.

*Ali, A., R.D. Baggs, and J.P. Stewart. 1981. Susceptibility of some Florida Chironomids and mosquitoes to various formulation of Bacillus thuringiensis serovar. Israelensis. J. Econ. Entomol. 74:672-677.

*Ali, A., L.C. Barbato, F. Ceretti, S. Sella Sala, R. Riso, G. Marchese and F.D=Andrea. 1992. Efficacy of two temephos formulations against Chironomus salinarius (Diptera: Chironomidae) in the saltwater lagoon of Venice, Italy. J. Amer. Mosq. Control Assn. 8:353-356.

*Ali, A. and J. Lord. 1980. Impact of experimental insect growth regulators on some nontarget aquatic invertebrates. Mosq. News. 40:564-

Ali, A. and M.S. Mulla 1976. Substrate type as a factor influencing spatial distribution of chironomid midges in an urban flood control channel system. Environ. Entomol. 5:631- 636.

Ali, A. and M.S. Mulla. 1976. Insecticidal control of chironomid midges in the Santa Ana River water spreading system, Orange County, California. J. Econ. Entomol. 69:509-513.

*Ali, A. and M.S. Mulla. 1977. Chemical control of nuisance midges in the Santa Ana River Basin, Southern California. J. Econ. Entomol. 70:191-195.

@Ali, A. and M.S. Mulla. 1978. Effects of chironomid larvicides and diflubenzuron on nontarget invertebrates in residential-recreational lakes. Environ. Entomol. 71:21- 27.

Ali, A. and M.S. Mulla. 1978. Declining field efficacy of chlorpyrifos against chironomid midges and laboratory evaluation of substitute larvicides. J. Econ. Entomol. 71:778- 782.

Ali, A. and M.S. Mulla. 1978. Chironomid population changes in an intermittent water spreading system. Mosq. News 38:386-392.

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*Anderson, L.M., J.H. Nelson, C. Thies, and M.V. Meisch. 1983. Evaluation of a controlled- release silicate formulation of temephos against Aedes aegypti larvae (Diptera: Culicidae) in rice field plots. J. Med. Entomol. 20:325-329.

Arias, M.R. and M.S. Mulla. 1975. Postemergence effects of two insect growth regulators on the mosquito Culex tarsalis (Diptera: Culicidae). J. Med. Entomol. 12: 317.

*Axtell, R.C. 1979. Principles of integrated pest management (IPM) in relation to mosquito control. Mosq. News 39: 709-718.

Axtell, R.C. (Ed.) 1974. Training manual for mosquito and biting fly control in coastal areas. UNC Sea Grant Publ. UNC-SG-74-08. 249pp. [Acc. No. COM-74- 1128/AS, Nat. Tech. Inf. Serv., Springfield, VA]

@Back, C., J. Boisvert, J.O. Lacoursiere, and G. Charpentier. 1985. High-dosage treatment of a Quebec stream with Bacillus thuringiensis serovar israelensis: efficacy against black fly larvae (Diptera: Simuliidae) and impact on non-target insects. Can. Entomol. 117:1523- 1534.

*Balcomb, R., R. Stevens and C. Bowen. 1984. Toxicity of 16 granular insecticides to wild- caught songbirds. Bull. Environ. Contam. Toxicol. 33:302-307.

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*Breaud, T.P., J.E. Farlow, C.D. Steelman and P.E. Schilling. 1977. Effects of the insect growth regulator methoprene on natural populations of aquatic organisms in Louisiana intermediate marsh habitats. Mosq. News 37:704-712.

*Brooks, G.D., H.F. Schoof and E.A. Smith. 1966. Evaluation of five formulations of Abate against Aedes aegypti, Savannah, Georgia, 1965. Mosq. News 26:580-582.

Brown, A.W.A. 1972. The ecological implications of insecticide usage in malaria programs. Am. J. Trop. Med. Hyg. 21:829-834.

Brown, P.W. and M.L. Hunter, Jr. 1984/85. Potential effects of insecticides on the survival of dabbling duck brooks. J. Minn. Academy Science 50:41-45.

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Busch, D.A. 1975. The failure of temephos and chlorpyrifos to affect the short-term productivity of (Spartina alterniflora) and (S. patens) on a New Jersey salt marsh. Proc. Ann. Meet. N.J. Mosq. Control Assoc. 62:50-55.

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*Campbell, B.C. and R.F. Denno. 1976. The effect of Temephos and Chlorpyrifos on the aquatic insect community of a New Jersey salt marsh. Environ. Entomol. 5:477-483. *Carey, W.F. 1974. Initial studies of Abate in a salt-marsh ecosystem: chemical studies. Proc., N.J. Mosq. Exterm. Assoc. 61:129-137.

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Carlson, D.B. and R.R. Vigliano. 1985. The effects of two different water management regimes on flooding and mosquito production in a salt marsh impoundment. J. Amer. Mosq. Control Assn. 1:203-211.

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@Case, T.J. and R.K. Washino. 1978. Effects of growth regulator methoprene on (Culex tarsalis) and non-target organisms in California rice fields. Mosq. News 38:191- 196.

*Catts, E.P., Jr., F.H. Lesser, R.F. Darsie, Jr., O. Florschutz, and E.E. Tindall. 1963. Wildlife usage and mosquito production on impounded tidal marshes in Delaware, 1956- 1962. Trans. 29th N. Am. Wildl. Res. Conf. 129-

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*Center for Disease Control. 1973. Public Health Pesticides. Technical Development Laboratories, Malaria Program, Center for Disease Control, Public Health Service, USDA. Reprint No. 530. 21pp.

@Chambers, H. And D.L. Fabacher. 1972. Midge larvae control in commercial catfish ponds: toxicity of AbateR to channel catfish (Ictularus punctatus). Proc. Annu. Conf. Southeast. Assoc. Game Fish Comm. 26:399-401. (Abstract)

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Coats, J.R. 1994. Risks from natural versus synthetic insecticides. Ann. Rev. of Entomol. 39:489-515.

*Colbo, M.H. and A.H. Undeen. 1980. Effect of Bacillus thuringiensis var. israelensis on nontarget insects in stream trials for control of Simuliidae. Mosq. News 40:368-371.

Cope, O.B. 1965. The effects of pesticides on fish and wildlife. U.S. Department of the Interior, Fish & Wildlife Service Circular 226:51-56.

@Costlow, J.D., Jr. 1977. The effect of juvenile hormone mimics on development of the mud crab Rhithropanopeus harrisii (Gould). In Physiological Responses of marine biota to pollutants. F.J. Thurberg and W.B. Vernberg (eds.) Academic Press, NY, pp. 439-457.

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*Dale, P.E.R. and K. Hulsman. 1990. A critical review of salt marsh management methods for mosquito control. Review in Aquatic Sciences 3:281-311.

*Dale, W.E., J.W. Miles and G.O. Guerrant. 1974. Monitoring of residues of Abate in streams treated for Similium control. IN Environmental Quality and Safety Supplement, Vol. III, Pesticides. Pp 780-783.

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*Dame, D.D. and K.E. Savage. 1981. Assessment of industrial formulations of Bacillus thuringiensis var israelensis. Mosq. News 41:540-546.

@Daorai, A. and R.E. Menzer. 1977. Behavior or Abate in microorganisms isolated from polluted water. Arch. Environ. Contamin. Toxicol. 5:229-240.

@Darwazeh, H.A. and M.S. Mulla. 1974. Toxicity of herbicides and mosquito larvicides to the mosquito fish (Gambusia affinis). Mosq. News 34:214-218.

@Darwazeh, H.A. and M.S. Mulla. 1981. Encapsulated formulations of primiphos- methyl against mosquito larvae and some nontarget organisms. ?? 49:90-94.

@Darwazeh, H.A. and M.S. Mulla. 1981. Pyrethrin tossits against mosquito larvae and their effects on mosquito fish and selected nontarget organisms. Mosq. News 41:650-655.

*Dash, A.P. and M.R. Ranjit. 1992. Comparative efficacy of aphid extracts and some juvenoids against the development of mosquitoes. J. Amer. Mosq. Control Assn. 8:247-251.

@Davison, K.L. 1976. Carbon-14 distribution and elimination in chickens given methoprene-14C. J. Agric. Food Chem. 24:641-643.

DeBord, D.V., G.A. Carlson and R.C. Axtell. 1975. Demand for and cost of coastal salt marsh mosquito abatement. North Caroling Agric. Expt. Sta. Tech. Bull. 232. 85pp.

*Denno, R.F. 1974. Initial studies of Abate in a salt-marsh ecosystem: sampling the nontarget component. Proc. N.J. Mosq. Exterm. Assn. 61:138-144.

@Derby, S.B. and E. Ruber. 1971. Primary production: depression of oxygen evolution in algal cultures by organophosphorous insecticides. Bull. Environ. Contam. Toxicol. 5:553-558.

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APPENDIX I. Environmental Effects of Mosquito Control

ENVIRONMENTAL EFFECTS OF MOSQUITO CONTROL (USFWS, 2004)

This paper provides a summary of the potential impacts of mosquito control practices. This is not intended to discount the important role that mosquitoes play in the transmission of disease, nor is it intended to diminish the role that mosquito control can play in reducing the incidence of such vector- borne disease in humans. The first part discusses the ecological role of mosquitoes in the environment, aside from that as vectors of disease. The second part addresses the potential impacts of mosquito control pesticides to nontarget organisms and communities.

1. The Ecological Role of Mosquitoes

Mosquitoes are most often associated with their roles as vertebrate ectoparasites and vectors of disease- causing microorganisms. However, do mosquitoes provide any Abeneficial@ role in the environment, or are they, as Spielman and D=Antonio (2001) claim Aself-serving@ and of Ano purpose other than to perpetuate her species@?

Mosquitoes As Prey

Larvae. Mosquitoes have evolved to use a wide variety of both permanent and temporary aquatic habitats for larval development. There are nearly as many habitats for mosquito larvae as there are types of lentic water bodies. For purposes of this discussion, mosquitoes will be divided into those that develop in ephemeral water bodies and those that develop in permanent to semi-permanent water.

The evolution of a drought- and sometimes freeze-resistant egg has allowed certain species of mosquitoesCthe most common in the genera Aedes, Ochlerotatus, and PsorophoraCto colonize a wide variety of ephemeral habitats large and small, from the tropics to sub-arctic zones. These mosquitoes lay eggs in dry or moist areas that will flood later. This strategy has at least two advantages: 1) the recently flooded detritus provides a nutrient-rich and abundant source of food for developing larvae; and 2) in many habitats there is a lag time before invertebrate predators colonize these temporary water bodies, allowing the larvae to develop in relatively predator-free environments. In most ephemeral habitats, mosquito eggs will hatch within hours of being flooded, often in very large numbers. In many of these habitats, such as summer flood pools and salt marshes, colonization by invertebrate predators occurs from highly mobile insects like dragonflies, beetles, and backswimmers that fly from more permanent bodies of water. Although some predators will arrive relatively quickly, it can take several days to weeks for an invertebrate predator community to become established. During the summer, a floodwater mosquito brood can develop from egg to adult in a week, and thereby mostly escape predation by these colonizing invertebrates.

In unpredictably flooded ephemeral habitats such as summer flood pools and storm-flooded salt marshes, there are few predators that have been identified to rely principally on mosquito larvae

as a source of food. The unreliable nature of mosquito larvae as prey in these habitats prevents the development of any close predator-prey relationship unless the predator shares diapausing strategies similar to those of floodwater mosquitoes. The only predators in these habitats that rely on mosquito larvae for prey are other mosquitoes. A few species of Psorophora mosquitoes have larvae that are predatory in late instar stages. These species are generally found in summer flood pools. Although there are few predators that specialize on mosquito larvae in these habitats, generalist predators such as beetles (larvae and adults), backswimmers, and some odonates (damselflies and dragonflies) will take advantage of the temporary abundance of mosquitoes if the timing of arrival into the habitats coincides with the presence of mosquito larvae.

Some ephemeral aquatic habitats, however, have flooding regimes that are more predictable. In at least two of these habitats, vernal pools and treeholes, we see the development of very close predator-prey relationships with mosquito larvae. In treeholes, species of another mosquito, Toxorhynchites, have evolved as predators of other treehole-dwelling mosquito larvae. Vernal pools in northern temperate regions predictably flood to their maximum extent in the early spring from rain and snowmelt, and this triggers a hatch of one or more species of Ochlerotatus mosquitoes. These are usually univoltine (single generation) species that laid eggs in the dry pool basin the previous summer. Hatching of mosquito larvae in vernal pools often occurs when water temperatures are still well below 10C, with few predators active in such cold environments. The predators present at this time of the year are generally those that share similar overwintering strategies with mosquitoes, such as cyclopoid copepods (e.g., Macrocyclops) and a few species of beetles. Some species of predaceous diving beetles (family Dytiscidae) in the genus Agabus have evolved a diapausing strategy that closely resembles that of the Ochlerotatus mosquitoes. Unlike most dytiscid beetles, these species have drought- and freeze-resistant eggs that are laid in the dry basin the previous summer and hatch in the early spring concurrently with mosquitoes. The beetle larvae are active in the cold water and appear to feed primarily on mosquito larvae and pupae (Nilsson and Soederstroem 1988; Higgins and Merritt 1999). The predictable abundance of mosquitoes and general paucity of other potential prey species during the early spring in these pools has probably contributed to this specialization. Other predators in vernal pools will feed opportunistically on mosquito larvae. Some species of dragonflies and damselflies (Odonata, primarily Sympetrum and Lestes) have also evolved drought- and freeze- resistant eggs, but hatch later in the spring. Colonizing species of backswimmers (Notonectidae), water striders (Gerridae), and water beetles (Hydrophilidae and Dytiscidae) will feed on late- instar mosquito larvae and pupae, but are considered generalist predators (Higgins and Merritt 1999).

Mosquitoes that require water for oviposition include the common genera Culex and Anopheles. These mosquitoes colonize permanent to semi-permanent bodies of water, laying eggs on the surface. In many natural bodies of water, the larvae of these species must develop in the presence of an oftentimes-diverse invertebrate predator community. The co-occurrence of mosquito larvae and predatory invertebrates is more predictable in these habitats, but the diversity of other potential prey species may preclude the development of specialized predator- prey relationships. Potential invertebrate predators in these habitats include: backswimmers, water striders, giant water bugs (Belostomatidae), water measurers (Hydrometridae), adult and

larval beetles (Dytiscidae, Hydrophilidae, Gyrinidae), many species of damselflies and dragonflies (Odonata), phantom midge larvae (Chaoboridae), and even copepods and flatworms. Although all of these predators can be considered generalists with regard to prey consumption, experimental evidence suggests that mosquito larvae, when available, are a preferred prey of some species (Helgen 1989; Urabe et al. 1990; Robert and Venkatesan 1997; Safurabi and Madani 1999).

Adults. Like other aquatic insects with terrestrial adult stages, mosquitoes provide a link between aquatic and terrestrial ecosystems as they convert detritus and aquatic microbial biomass into flying insect biomass. Most adult mosquitoes are relatively short lived. The probability of daily survival for adult mosquitoes, an important factor in disease transmission, varies among species and habitats. Daily survival probabilities usually range from 0.6-0.9, with much of the mortality coming from predation (Service 1993). Mosquitoes are fed upon by a variety of invertebrate predators, including spiders (Strickman et al. 1997; Fox 1998) and odonates (Sukhacheva 1996), although there are no known specialist predators that prey exclusively on mosquitoes. Vertebrate predators include insectivorous birds and bats (Zinn and Humphrey 1981), although mosquitoes often account for only a small percentage of the total biomass consumed. Consumption of mosquitoes by the Indiana bat, Myotis sodalis, for example, accounted for up to 6.6 percent of the total diet (Kurta and Whitaker 1998). The apparent absence of any specialized predator-prey relationships among adult mosquitoes and predators, however, does not necessarily discount the contribution of mosquitoes to the diet of a wide variety of generalized predators.

Other Ecological Roles of Mosquitoes

Mosquito larvae may feed by one or more of several different mechanisms. They may filter-feed, graze microbial biofilms, or even shred detritus (Merritt et al. 1992). In this sense, mosquitoes are a component of a functioning wetland ecosystem, processing detritus and aquatic microbes, and eventually providing a link between aquatic and terrestrial systems when they emerge.

Most adult mosquitoes require sugar meals during their lifetimes as a source of energy. The primary sources of sugars consumed by mosquitoes are nectar from flowers and honeydew excreted by aphids (Foster 1995). Both male and female mosquitoes frequently take nectar meals from flowers, but are they important as pollinators? Due to their small size and the limited probing abilities of the proboscis, mosquitoes are limited to feeding on nectar sources within flowers that have shallow or flat corollas. Unlike relatively large pollinators like bees and butterflies, mosquitoes can nectar feed efficiently without coming into contact with pollen-coated stamens. Thus, although they may transfer some pollen during the course of acquiring a meal of nectar, mosquitoes are probably not important pollinators in general (Foster 1995). A documented exception to this occurs in the subarctic where mosquitoes are significant pollinators of many plants (Kevan 1972).

The impact of reducing the density of mosquitoes in aquatic or terrestrial systems has not been studied. Generalist predators probably switch to alternate prey, which in turn may be impacted by the increased predation. The few specialist predators of mosquito larvae may be impacted the

greatest due to the lack of alternate prey and/or the inability of such predators to uncouple from a closely evolved predator-prey relationship. 2. Nontarget Effects of Mosquito Control Pesticides

Mosquito control pesticides can be categorized into three groups: larvicides, adulticides, and water surface films (for controlling mosquito larvae and pupae). Compared with other forms of pest control, there are relatively few pesticides available within each of these categories, and all differ with regard to efficacy and effects on nontarget organisms.

Larvicides

Bacillus thuringiensis var. israelensis (Bti). Like other varieties of the natural soil bacterium, Bacillus thuringiensis (Bt), Bti is a stomach poison that must be ingested by the larval form of the insect in order to be effective. Bt contains crystalline structures containing protein endotoxins that are activated in the alkaline conditions of an insect=s gut. These toxins attach to specific receptor sites on the gut wall and, when activated, destroy the lining of the gut and eventually kill the insect. The toxicity of Bt to an insect is directly related to the specificity of the toxin and the receptor sites. Without the proper receptor sites, the Bt will simply pass harmlessly through the insect=s gut. Several varieties of Bt have been discovered and identified by the specificity of the endotoxins to certain insect orders. Bacillus thuringiensis var. kurstaki, for example, contains toxins that are specific to lepidopterans (butterflies and moths), while Bti is specific only to certain primitive dipterans (flies), particularly mosquitoes, black flies, and some chironomid midges. Bti is not known to be directly toxic to nondipteran insects.

Because Bti must be ingested to kill mosquitoes, it is much more effective on first-, second-, and early third-instar larvae than on late third and fourth instars since the earlier instars feed at a faster rate (fourth instar larvae feed very little). The pesticide is completely ineffective on pupae because they do not feed at all. Formulated products may be granular or liquid, and potency is expressed in International Toxicity Units (ITU), usually ranging from 200-1200 ITU. The concentrations of Bti in water necessary to kill mosquito larvae vary with environmental conditions, but are generally 0.05-0.10 ppm. Higher concentrations (0.1->0.5 ppm) of Bti are necessary when there is a high amount of organic material in the water, late-third and early fourth instar larvae predominate, larval mosquito density is high, or water temperature is low (Nayar et al. 1999). Operationally, Bti is applied within a range of volume or weight of formulated product per acre as recommended on the pesticide label, with the goal to achieve an effective concentration. The label recommended range of application rates under most conditions varies by a factor of 4 for most formulations (e.g., for granular formulations, 2.72-11.12 kg/ha (2.5-10 lb/acre)). For later instar larvae and water with a high organic content, higher application rates are recommended that may reach 8 times the lowest rate (e.g., for granular formulations, the higher rate is 11.1-22.5 kg/ha (10- 20 lb/acre)). Mosquito control agencies use the recommended label rates, along with previous experience, to administer an effective dose. Typical application rates are often in the mid- to upper values of the normal ranges recommended on the labels (Abbott Laboratories 1999). Because water depths even within a single wetland can vary greatly, field concentrations of Bti can vary widely, especially when the pesticide is applied aerially.

Efficacy is monitored by post-application reductions in mosquito larval density, but the actual concentration of Bti following an application is not measured. Thus, an insufficient concentration of Bti can be detected by low mortality of mosquito larvae, but an overdose (i.e., a concentration greater than necessary to kill mosquito larvae) of the pesticide is rarely monitored for.

The issue of Bti concentration is important with regard to impacts on nontarget organisms. Of particular concern is the potential for Bti to kill midge larvae (family Chironomidae). Chironomid (non-biting midge) larvae are often the most abundant aquatic insect in wetland environments and form a significant portion of the food base for other wildlife (Batzer et al. 1993; Cooper and Anderson 1996; Cox et al. 1998). Negative impacts on chironomid density/biomass could have deleterious effects on wetland/wildlife food webs and could also lower biodiversity.

The potential for Bti to impact chironomid populations depends on the fate and availability of the pesticide, the ingestion of the pesticide, and the presence and number of specific receptor sites in the insect gut for the toxins (as discussed above). Fate and availability encompass both the initial dose/concentration and the fate of the pesticide in the aquatic environment. Chironomid larvae live primarily in the benthos of wetlands. Mosquito larvae ingest Bti primarily within the water column, but Bti readily adheres to suspended particulate matter and settles to the benthos (Yousten et al. 1992).

Ingestion of Bti by chironomid larvae depends primarily on the feeding mechanism. The family Chironomidae is a relatively large group, with nearly 1,000 species identified for North America (Merritt and Cummins 1996). This family encompasses a variety of feeding strategies: filter- feeders, collector- gatherers, scrapers, shredders, and even predators. Filter-feeding larvae are more likely to ingest Bti than larvae with other feeding strategies (Pont et al. 1999).

Chironomid larvae appear to possess mid-gut receptor sites for Bti endotoxins similar to those in mosquito larvae, and exhibit similar histopathological changes in the gut lining that lead to death of the insects when exposed to lethal concentrations of the pesticide (Yiallouros et al. 1999). There are, however, differences in the susceptibility of midge larvae to Bti at the subfamily level and among larval instars. In general, larvae in the subfamily Chironominae (Tribes Chironomini and Tanytarsini) are more susceptible to Bti than larvae of other subfamilies (Yiallouros et al. 1999) (Pont et al. 1999) (Ali 1981). Also, early-instar larvae are much more susceptible to Bti than later instars (Ali et al. 1981; Charbonneau et al. 1994).

There have been a number of laboratory and field studies examining the toxicity of Bti to chironomid larvae (Boisvert and Boisvert 2000). There have been many different formulations and potencies of Bti products used in these studies, and in many cases actual concentrations of Bti within the water were not measured. Also, differences in the species and instar of the chironomid larvae used (sometimes not specified), and in the environmental conditions of the field experiments make direct comparisons among the studies difficult. Most field studies examined the nontarget effects from a single application of Bti and did not address the potential long-term impacts from repeated applications over a season or over several seasons.

It is clear that in laboratory studies Bti is lethally toxic to some species of chironomid larvae at concentrations expected for mosquito control. Charbonneau et al. (1994) determined an EC50 (the concentration required to cause an effect in 50 percent of the test population) of 0.20 ppm for Chironomus riparius (fourth instar?), and the toxicity of Bti to earlier instars was over two orders of magnitude greater. Similarly, Ali et al. (1981) found the LC50 (the concentration required to kill 50 percent of the test population) for first-instar Glyptotendipes paripes (0.034 ppm) to be over two orders of magnitude lower than the LC50 for third instar larvae (8.31 ppm).

Charbonneau et al. (1994) studied the effects of Bti on chironomid larvae in the laboratory and the field. Laboratory toxicity tests on Chironominae larvae (the most susceptible subfamily) demonstrated up to 100 percent mortality at label-recommended rates, but the toxicity of Bti to chironomids was influenced by several environmental factors. Factors that lowered toxicity to chironomids included higher water temperature, greater water depth, organic matter, and coverage by macrophytes. Field enclosure tests with Bti applied at 5.6 kg/ha (5 lb/acre) failed to demonstrate any pesticide effects on midge larvae within the enclosures, leading the authors to conclude that environmental factors reduce the toxicity of Bti to chironomids in the field. However, mortality of nontarget organisms within the enclosures was measured after 48 hours. Apparent effects of Bti on chironomids may not be detectable for 5-7 days post application (Ali 1981; Lacey and Mulla 1990; Pont et al. 1999). Also, because early instar larvae are much more susceptible to Bti, first and second instars would likely exhibit the greatest mortality. The 575 m mesh used to sample benthic invertebrates in the field tests of the Charbonneau et al. (1994) study, however, was too large to effectively sample first- and some second-instars. Thus, the conclusions regarding the field component of this study must be viewed with caution.

There is some evidence from field studies in which negative impacts to chironomid larvae were observed that such impacts are relatively short-lived (e.g., Miura et al. 1980). In most of these studies, however, it is not clear if the rebounding densities of midge larvae represent the same species or even the same subfamily that was initially reduced by the pesticide. Furthermore, population-level impacts to species from repeated applications over a season were usually not addressed. Although many species of chironomids are capable of producing several generations per year and could re-colonize a treated wetland relatively quickly, other species have only one generation per year and therefore would not be able to re-colonize until the following year. The ability of Bti-susceptible species to re-colonize a wetland following pesticide treatment would also depend on 1) the frequency of Bti applications, 2) the extent of Bti treatments within the wetland, and 3) the extent of Bti applications in the surrounding landscape. Widespread larviciding with Bti would provide few refugia for re-colonizing source populations of susceptible species.

In a study that examined population-level impacts to chironomids from a single application of Bti at a mosquito control rate, investigators showed that, while there was no statistical difference in the number of emerging adult chironomids between control and treatment enclosures, the species composition was different (Pont et al. 1999). Species sensitive to Bti (Tanytarsus horni, T. fimbriatus, and Microchironomus deribae) were 24-54 percent less abundant in enclosures treated at mosquito control rates than in control enclosures, while a less sensitive species

(Polypedilium nubifer) was over 200 percent more abundant in the treated enclosure versus control. Higher application rates resulted in greater reductions of the Bti-sensitive species. This suggests that as Bti-sensitive chironomid larvae are killed by the pesticide, less sensitive species may thrive as they are released from competition (Pont et al. 1999). Thus, although chironomid larval numbers often appear to rebound after a treatment with Bti, this may be indicative of a shift in the species composition of the community, with species less sensitive to Bti replacing the sensitive species. It is unknown how or if such a shift would affect food web dynamics, but biodiversity would be lowered.

There is only one published study that examined the long-term, nontarget effects of Bti (Hershey et al. 1998; Niemi et al. 1999). In this study conducted in Minnesota, 27 wetlands were sampled for macroinvertebrates over a 6-year period. All wetlands were sampled for 3 pre-treatment years and randomly assigned to 3 treatment groups: Bti, methoprene (see discussion below), and an untreated control group. The wetlands were sampled for 3 treatment years. Bti was applied to wetlands in a granular formulation at the rate of approximately 11.1 kg/ha (10 lb/ac), which represents the high end of the normal label-recommended application range. Bti was applied to each treatment wetland 6 times per year at intervals of 3 weeks or after rainfall of >1.25 cm, whichever came first (Niemi et al. 1999). Although this frequency of application is high, it is within the range that could occur from operational mosquito control.

After the first year of treatment, no differences in macroinvertebrate density, biomass, or community composition (richness of genera) among the treatments were observed (Hershey et al. 1998). However, in the second and third years of treatment, highly significant differences were observed in the two treatment groups compared to control. Chironomid larvae were significantly impacted by Bti treatments, with reductions in density of 66 percent and 84 percent for the second and third years of the study, respectively, compared to densities in control wetlands (Hershey et al. 1998). Significant declines in other nematoceran (primitive) dipteran larvae were also observed during the last two years of the study. There were also declines in macroinvertebrate predator densities in the Bti treated wetlands that the authors interpret as indirect effects from the reduction in a prey base dominated by chironomid larvae (Hershey et al. 1998; Niemi et al. 1999).

In summary, there is clear evidence from both laboratory and field studies that Bti can kill some chironomid larvae. Species in the subfamily Chironominae are apparently the most susceptible to direct toxicity; other subfamilies exhibit little mortality at mosquito control rates. Even within the subfamily Chironominae there are apparent differences among in susceptibility to Bti, relating perhaps to feeding mode (Pont et al. 1999). Within susceptible species, toxicity is greatest to early instars. Lethal concentrations of Bti are orders of magnitude lower for early versus late instars, and well within the concentrations expected from operational mosquito control. There is evidence that environmental conditions such as temperature, organic content of the water, vegetation, and density of larvae can ameliorate some of the potential negative impacts to chironomid larvae (Charbonneau et al. 1994), although field experiments designed to test this may be suspect.

The only long-term study on the nontarget effects of Bti for mosquito control demonstrated significant adverse effects on the chironomid community of treated wetlands, and this translated into numerous significant negative effects within the food web (Hershey et al. 1998; Niemi et al. 1999). The intensity of Bti applications used in this study, both the application rate and the frequency of applications, would represent the high end of those that would normally occur for operational mosquito control. In addition, entire wetlands were treated, which may or may not occur with aerial applications of Bti. Thus, the Minnesota study may represent a Aworst-case scenario@ of potential mosquito control operations, but it has generated the only data available on the long-term nontarget effects from Bti. Studies that examine nontarget effects of Bti from a single application or even within a single season may not be adequate to detect potential long- term impacts from pesticide use (Hershey et al. 1995).

There is also evidence that application rate can have a profound effect on impacts to chironomids from Bti (Rodcharoen et al. 1991). Because application rates of Bti for mosquito control can vary by a factor of 8, field concentrations of the pesticide can reach levels that are toxic to chironomid larvae, yet are still within the pesticide label directions. In addition, there are no label restrictions on the number of applications of Bti to any one area. Economic considerations may preclude regular applications at the highest label rate, yet even at lower rates, adverse impacts to chironomid midge larvae have been demonstrated (Miura et al. 1980; Ali 1981; Ali et al. 1981).

Bacillus sphaericus

Bacillus sphaericus (Bsph) is a naturally-occurring soil bacterium similar to Bti, and has been developed as a commercially-available mosquito larvicide since the early 1990s. Like Bti, it releases a protein endotoxin in the alkaline gut of larval mosquitoes that attaches to specific receptor sites of susceptible species. This endotoxin dissolves the lining of the gut wall and eventually kills the larva. Unlike Bti, Bsph has only one endotoxin (Bti has two or more). Also, unlike Bti, Bsph is very effective in water with a high organic content, and is therefore often used in such habitats for control of Culex mosquitoes. Bsph is also capable of Acycling@ in the aquatic environment, meaning it can retain its larvicidal properties after passing through the gut of a mosquito andCunlike BtiCprovide effective mosquito control for weeks after a single application. Bsph, however, is not effective on all species of mosquitoes.

Because Bsph is a more recently developed larvicide than Bti, there are fewer studies that have examined the nontarget effects of this pesticide. The data available, however, indicate a high degree of specificity of Bshp for mosquitoes, with no demonstrated toxicity to chironomid larvae at any mosquito control application rate (Mulla et al. 1984; Ali and Nayar 1986; Lacey and Mulla 1990; Rodcharoen et al. 1991). This high specificity to some mosquito species and low toxicity to chironomid larvae is probably the consequence of the one endotoxin contained with the Bsph spore. Unfortunately, this also makes the development of resistance to this pesticide more likely if this pesticide becomes widely and frequently used.

Methoprene

Methoprene is a synthetic mimic of a naturally produced insect hormone, juvenile hormone (JH). All insects produce JH in the larval stages, with the highest levels occurring in the insect=s early developmental stages. As an insect reaches its final stage of larval development, the level of JH is very low. This low level of JH triggers the development of adult characteristics. When an insect is exposed to methoprene, a hormonal imbalance in the development of the insect results, and it fails to properly mature into an adult. The insect eventually dies in the pupal stage. The most susceptible stages of development to methoprene are the later instars (for mosquitoes, third and fourth instars). In mosquito control applications, methoprene is applied directly to the larval breeding habitat. Larvae will continue to feed and may reach the pupal stage, but they will not emerge as adults. Methoprene is completely ineffective on mosquito pupae and adults. It is available in several formulations: liquid, granular, pellet, and briquet. There are several microencapsulated and extended-release formulations that remain effective for up to 150 days.

The amount of methoprene necessary for mosquito control is < 1.0 part per billion (ppb). The initial concentrations of methoprene when applied to aquatic habitats may reach 4-10 ppb, but residual concentrations are approximately 0.2 ppb (Ross et al. 1994). Once released into the aquatic environment, it is non-persistent, with a half-life of about 30-40 hours. Micro- encapsulated and extended-release formulations will, of course, be present in the water longer as the pesticide is slowly released over time, 7- 150 days, depending on the formulation. In field applications, efficacy is determined only by an observed inhibition of emergence of adults, since larvae are not directly killed by the pesticide.

Because methoprene is a JH mimic and all insects produce JH, there is concern about potential adverse impacts to nontarget aquatic insects when this pesticide is used for mosquito control. As with Bti, there is particular concern regarding potential negative impacts to chironomid larvae due to their importance in food webs. As with any pesticide, toxicity is a factor of dose plus exposure. At mosquito control application rates, methoprene is present in the water at very small concentrations (4-10 ppb, initially). With regard to exposure, chironomid larvae occur primarily in the benthos, either within the sediments and/or within cases constructed of silk and detritus. Thus, there may be differences with regard to exposure to methoprene between chironomid and mosquito larvae, the latter occurring primarily in the water column.

The published literature on the impacts of methoprene to chironomids is not as extensive as that for Bti. However, there is evidence for potential toxicity to chironomid and other aquatic invertebrates from methoprene treatments. Some early experiments indicated approximately 50 percent mortality of Chironomus stigmaterus (Chironomidae) and 70 percent of Brachydeutera argentata (Diptera: Ephidridae) larvae when exposed to 0.01 ppm of technical grade methoprene (Miura and Takahashi 1973). Mulla et al. (Mulla et al. 1974) noted up to 100 percent inhibition of emergence for some midge species, although the lowest concentration tested was 0.1 ppm. Breaud et al. (1977) observed reductions in several aquatic invertebrate taxa, including chironomids, after six applications of methoprene over an 18-month period in a Louisiana marsh. The application rate in this latter study was 0.028 kg/ha of active ingredient, although the formulation was not specified (Breaud et al. 1977).

In testing different formulations of methoprene against chironomids in experimental ponds, Ali (1991a) found that sustained-released formulations inhibited emergence of midges by 38-98 percent, in some cases for up to 7 weeks. A liquid, microencapsulated formulation applied at mosquito control rates resulted in a 60 percent inhibition of emergence in the tribe Chironomini for 14 days post-treatment. A pelletized, sustained-release (30 days) formulation applied at mosquito control rates inhibited all chironomid emergence by 64-98 percent for 7 weeks. A briquet formulation (30 days sustained-release) produced 38- 98 percent inhibition of all chironomids for 7 weeks. The granular formulation applied at the high end of mosquito control rates reduced chironomid emergence by 61-87 percent (Ali 1991a).

In the multi-year Minnesota study cited above, a 3-week sustained-release, granular formulation of methoprene was applied to treatment wetlands at a label-recommended rate of 5-10 kg/ha (Hershey et al. 1998; Niemi et al. 1999). The pesticide was applied six times per season at 3- week intervals. The impacts from methoprene in this study were very similar to those observed for Bti. Negative impacts were not observed until the second and third years of treatment. In those years, significant declines in aquatic insect density and biomass were detected in methoprene-treated wetlands compared to controls. Total insect biomass was 70 percent and 81 percent lower in the second and third years of treatment, respectively, than in control wetlands (Hershey et al. 1998). Reductions were observed across many insect taxa, including predators and non-predators, suggesting direct (pesticide) and indirect (food web) effects from methoprene treatments (Hershey et al. 1998).

Although the application rate of methoprene used in the Minnesota study was well within operational rates used in mosquito control, the frequency of application exceeded what would probably occur under most field situations. Using a 3-week sustained release formulation and applying that every 3 weeks ensured a nearly constant exposure of methoprene to aquatic invertebrates in the treated wetlands throughout the season. Under such a scenario, it is unlikely that most impacted invertebrate populations would be able to re-colonize the wetlands during the treatments. However, this does not discount the conclusion that nontarget aquatic invertebrates were indeed impacted by methoprene at rates and concentrations used for mosquito control. Whether or not the observed food web effects would have been lessened under a more realistic pesticide application regime is debatable.

Studies of adverse impacts from methoprene on insect taxa other than chironomids are less conclusive. Because methoprene affects insect development and does not directly kill larvae, traditional toxicity testing over a few days is often inadequate when looking for potential impacts. Methoprene toxicity can only be observed at the point in which the immature insects reach (or fail to reach) adulthood. Thus, many published laboratory and field studies looking at nontarget impacts from methoprene were of insufficient duration to detect actual negative impacts (e.g., Miura and Takahashi 1973).

Breaud et al. (1977) observed adverse effects from methoprene on 14 aquatic invertebrate taxa, including Callibaetis sp. mayflies, odonates (dragonflies and damselflies), predaceous diving beetles, and chironomids. Negative impacts to Callibaetis mayflies from methoprene treatments have been observed by others (Steelman et al. 1975; Norland and Mulla 1975). Miura and

Takahashi (1973) did not observe any mortality on Callibaetis from methoprene in laboratory or field studies, but neither was of sufficient duration (48 hours and 1 week, respectively) to adequately detect developmental effects (Miura and Takahashi 1973). Pinkney et al. (2000) observed consistently lower numbers of mayflies emerging from methoprene-treated wetlands compared to controls, but these differences were not statistically significant (Pinkney et al. 2000).

There is evidence of methoprene impacts to non-insects as well. McKenney and Celestial (1996) noted significant reductions in number of young produced in mysid shrimp at 2 ppb (McKenney and Celestial 1996). Sub-lethal effects on the cladoceran, Daphnia magna, in the form of reduced fecundity, increased time to first brood, and reduced molt frequency have also been observed at concentrations < 0.1 ppb (Olmstead and LeBlanc 2001).

There has been speculation and some preliminary data to suggest that methoprene causes limb malformations in amphibians (La Clair et al. 1998). However, experiments with methoprene and its degradation products have failed to demonstrate developmental toxicity even at concentrations exceeding 100 times that expected for mosquito control (Ankley et al. 1998; Degitz et al. 2003). Therefore, current data do not support a role of methoprene in amphibian malformations.

In summary, there is evidence for significant adverse nontarget effects from methoprene even when applied at mosquito control rates. With regard to negative impacts to chironomid midges, there may be differences in susceptibility among species and differences depending on the formulation used. One study in particular suggested that methoprene formulations with short- term residual activity may have smaller impacts to chironomids (Ali 1991a). However, even the "ineffective" liquid formulation used in this study reduced emergence of Chironomini midges by 60 percent for two weeks. Certainly, not all midges will be affected by a single application of methoprene for mosquito control. However, the apparent differences in pesticide formulations, the varied susceptibility of species, and perhaps even the influence of some as- yet-undetermined environmental factors, make predicting the degree of any impacts nearly impossible.

Because methoprene does not immediately kill susceptible chironomid larvae, they are still available for predators. However, repeated applications of methoprene over a mosquito breeding season would eventually hinder recruitment as adults repeatedly fail to emerge (Hershey et al. 1998). Longer-term studies conducted over the course of a season or over multiple seasons are especially necessary for examining nontarget impacts from methoprene in order to detect potential impacts on longer-lived larvae (e.g., odonates, mayflies, and aquatic beetles) and to detect potential impacts to long- term recruitment. As was the case with Bti, the ability for a population to re-colonize a wetland following a methoprene treatment would depend on the intensity and frequency of applications at different spatial scales.

Temephos

Temephos is the only remaining organophosphate pesticide used for larval mosquito control. Like all organophosphate pesticides, it functions on the nervous system by inhibiting the production of acetylcholinesterase. Without this enzyme, nerves continue to fire, eventually resulting in death of the insect. Temephos is available in liquid or granular formulations that are

applied directly to aquatic breeding habitats of mosquitoes. Expected environmental concentrations of temephos in water are 20-35 ppb, but actual field concentrations can vary widely (Pierce et al. 1996). Temephos is not persistent, but can remain effective for 7-10 days (Fortin et al. 1987).

There have been many studies examining the adverse nontarget impacts of temephos. Many of these studies have documented significant negative impacts to a wide range of aquatic taxa, especially in freshwater wetlands. Temephos is very highly toxic to cladocerans (water fleas, e.g., Daphnia) at fractions of expected mosquito control concentrations (Fortin et al. 1987; Helgen et al. 1988). The U.S. Environmental Protection Agency (EPA) has determined an LC50 value of 0.01 ppb for Daphnia magna (EPA 1999), orders of magnitude lower than the expected environmental concentration of 20-35 ppb. The pesticide is highly toxic to chironomid larvae at or below mosquito control concentrations (Mulla and Khasawinah 1969; Iannacone and Alvarino 1998; Pinkney et al. 2000), although some researchers have documented only minimal effects on some species (Ali et al. 1978) (Ali 1991b). Temephos is especially toxic to larvae of the non- biting phantom midge, Chaoborus (Fales et al. 1968; Helgen et al. 1988; Pinkney et al. 2000). Temephos has also been found to be very toxic to potential mosquito predators such as odonates and backswimmers (Fales et al. 1968). Pinkney et al. (2000) reported significant reductions in insect diversity, richness, and density within temephos-treated experimental ponds, with significant declines in Ephemeroptera (mayflies), Odonata, Diptera, and Chironomidae compared to control ponds.

The effects of temephos on nontarget estuarine species are less studied. There are some data that suggest negative impacts from temephos are not as pronounced on estuarine species (Lawler et al. 1999b). However, there is evidence for toxicity to estuarine crustaceans from temephos at concentrations below those expected from field applications for mosquito control (Mortimer and Chapman 1995; Brown et al. 1996). Studies have also shown sublethal and indirect impacts of temephos on fiddler crabs (Ward and Busch 1976; Pinkney et al. 1999). In addition, at least some chironomid species in salt marsh habitats are susceptible to temephos (Ali et al. 1992).

Temephos has also been shown to be lethal to tadpoles of green frogs at concentrations < 10 ppb (Sparling et al. 1997).

It is clear that temephos is a much less specific larvicide compared to Bti and methoprene. Severe, negative impacts from temephos at mosquito control concentrations have been documented for a broad range of aquatic taxa in both freshwater and estuarine habitats, although some estuarine species are apparently more tolerant of the pesticide.

Surface Oils and Films

Surface oils and films are applied to mosquito breeding sites to kill mosquito larvae and pupae. The products create a barrier to the air-water interface and suffocate the insects, which require at least periodic contact with the water surface in order to obtain oxygen. The oils are mineral oil based and are effective for 3-5 days. Surface films are alcohol based and produce a monomolecular film over the water surface.

Both the oils and the films are potentially lethal to any aquatic insect that lives on the water surface or requires periodic contact with the air-water interface to obtain oxygen. Studies have demonstrated very significant negative impacts to water surface-dwelling insects from applications of oils (Mulla and Darwazeh 1981; Lawler et al. 1998).

Surface oils may also adversely impact wildlife by wetting the feathers of young waterfowl. This may be of particular concern at low temperatures when the oil could affect thermoregulation (Lawler et al. 1998).

Adulticides

All pesticides used to kill adult mosquitoes are broad-spectrum insecticides. The only selective aspect of these pesticides is in the manner in which they are applied. Most adulticides used currently are applied as ultra-low volume (ULV) sprays, meaning relatively small amounts are used (compared to some agricultural pesticides) and they are sprayed as very fine droplets (10-30 m in diameter). This small droplet size allows the spray to drift for a relatively longer period oftime compared to larger droplets, and the small size delivers an appropriate dose of the pesticide to kill an adult mosquito. Drift is a necessary component of adulticiding because these sprays are most effective on flying insects. For this reason, adulticide applications generally occur in the evening or early morning hours when the majority of mosquito species are most active. Adulticides may be applied by truck-mounted sprayers or applied aerially by helicopter or fixed- wing aircraft.

There are only two general classes of adulticides: organophosphates and pyrethroids. Both classes of pesticides work on the nervous system, although have different modes of action. Organophosphates are cholinesterase inhibitors while pyrethroids are sodium channel blockers. There are currently three organophosphate adulticides: malathion, naled, and fenthion, although fenthion is used only in a few counties in Florida and will be removed from the market in 2004. The most common pyrethroids are the synthetic pyrethroids, permethrin, resmethrin, and sumithrin. The pyrethroids are usually combined with the synergist piperonyl butoxide, which interferes with an insect's detoxifying mechanisms. None of these pesticides is persistent, with half-lives ranging from hours (naled) to several days (malathion and some pyrethroids).

Nontarget toxicity from adulticides may occur in either terrestrial or aquatic habitats as a result of deposition, runoff, inhalation, or ingestion. In general, pyrethroids have lower toxicity to terrestrial vertebrates than the organophosphates. With the exception of fenthion, which is highly toxic to birds, the application rates of the organophosphate adulticides are not likely to cause any direct mortality of vertebrates. Pyrethroids, although less toxic to birds and mammals, are very toxic to fish and aquatic invertebrates (Anderson 1989; Siegfried 1993; Milam et al. 2000). The actual toxicity of pyrethroids in aquatic habitats, however, is less than may be anticipated because of the propensity of these pesticides to adsorb to organic particles in the water (Hill et al. 1994). There are also data that indicate synthetic pyrethroid degradates have endocrine disrupting properties (Tyler et al. 2000).

In general, there are very few studies that have examined the nontarget effects of mosquito control adulticides. As all of these chemicals are broad-spectrum insecticides, they are potentially lethal to most insects. Yet there is a paucity of data available on the nontarget impacts to either terrestrial or aquatic invertebrates. There are data indicating the high toxicity of adulticides to honey bees (Taylor et al. 1987; Hagler et al. 1989; Pankiw and Jay 1992a; Pankiw and Jay 1992b), although the timing of adulticide applications in the evening can be expected to minimize these impacts.

Salvato (2001) examined the toxicity of naled, malathion, and non-synergized permethrin to 5 species of butterflies, including larval and adult stages. Naled and permethrin were found to be the most toxic to all life stages. The LD50 data presented for some larvae and adults coincide with that delivered by a single ULV droplet of 5-23 m, within the desired range for mosquito control (Salvato 2001). Mosquito control adulticiding has been identified as a likely contributing factor in the decline of several rare lepidopteran species in the Florida Keys (Calhoun et al. 2000; Salvato 2001).

All adulticides are very highly toxic to aquatic invertebrates in concentrations < 1 ppb (Milam et al. 2000). Because most adulticides can be applied over or near water when used for mosquito control, there are risks to aquatic invertebrates from direct deposition and runoff of the pesticides. However, very few field studies have been conducted that have examined the impacts to aquatic organisms from mosquito control adulticides. Jensen et al. (1999) failed to detect reductions in aquatic invertebrate abundance or biomass from truck-mounted applications of pyrethrin, permethrin, and malathion. However, the potentially most sensitive group of invertebrates, cladocerans (water fleas), were not sampled (Jensen et al. 1999). This could be important given that malathion residues of 6 ppb were recovered from water in the treatment areas during this study. This is several times the LC50 values of 0.69 ppb and 1.8 ppb of malathion for Simocephalis serrulatus and Daphnia magna, respectively (USEPA 2000), indicating that cladocerans would be at risk from applications of malathion for mosquito control. Declines in flying insect abundance were also observed during this study following pesticide applications, but the numbers quickly rebounded (Jensen et al. 1999).

As was the case with studies of nontarget impacts from larvicides, the limited numbers of studies on adulticide impacts all involve examining short-term effects, usually from a single application of a pesticide. It is difficult to extrapolate the results of short-term experiments into predictions of long-term impacts, whether the short-term studies detected impacts or not. In addition, mosquito control is most often conducted at a landscape level. Studies of impacts at such larger temporal and spatial scales are non- existent, and would be a challenge both scientifically and economically.

Biological Control

The mosquitofish, Gambusia affinis, has been used for decades as a biological control of mosquito larvae. These fish are effective in removing mosquito larvae from relatively small, closed, and artificial aquatic systems, such as backyard ponds. In more complicated natural systems however, Gambusia are not selective predators, and can adversely impact native

vertebrate and invertebrate communities (Rupp 1996). They can out-compete many native species of fish by feeding on eggs and fry, and they can actually reduce the density of natural invertebrate predators. There is also evidence that mosquitofish may cause direct and indirect impacts on tadpoles (Lawler et al. 1999a).

Summary/Conclusions

Mosquitoes are a natural component of many aquatic and terrestrial ecosystems. Like other aquatic insects with terrestrial adult stages, mosquitoes provide a link between aquatic and terrestrial habitats. Predation is probably the largest source of mortality for both larval and adult mosquitoes and, although there are relatively few predators that specialize on mosquitoes, these insects are fed upon by a wide variety of invertebrate and vertebrate predators. The impact of greatly reducing mosquito populations in aquatic and terrestrial ecosystems has not been studied.

Virtually every pesticide currently used to manage mosquito populations has the potential to adversely impact nontarget species. Widely used larvicides such as Bti and methoprene have been demonstrated to kill susceptible chironomid midge larvae, with experimental evidence suggesting that such population- level impacts may result in community-level food web effects. All adulticides are broad-spectrum insecticides that can potentially impact a wide variety of invertebrates and some vertebrates. The degree to which nontarget organisms or communities may be impacted by mosquito control pesticides is often difficult to predict because of differences in susceptibility among species, differences in toxicity of various formulated products, and basic knowledge gaps in toxicity data to certain species. An additional factor is the paucity of studies examining nontarget impacts of mosquito control at large spatial and temporal scales.

Organized mosquito control most often occurs at a landscape level such as a county or parish. When pesticides are applied to manage mosquito populations, it is often at multiple locations over relatively large spatial scales. Furthermore, pesticides may be applied to any given area multiple times in a season, year after year. The majority of nontarget mosquito control pesticide studies have examined impacts at much smaller temporal and spatial scales, such as one application in a single wetland. While these studies provide useful data, it is difficult to extrapolate the results of these small-scale experiments into predictions of impacts from much larger scale treatments.

References Cited

Abbott Laboratories. 1999. Technical Bulletin, Granular Formulations VectoBac. North Chicago, IL.

Ali, A. 1981. Bacillus thuringiensis Serovar. israelensis (ABG-6108) Against chironomids and Some nontarget aquatic invertebrates. Journal of Invertebrate Pathology 38: 264-272.

Ali, A. 1991a. Activity of new formulations of methoprene against midges (Diptera: Chironomidae) in experimental ponds. Journal of the American Mosquito Control Association 7: 616-620. Ali, A. 1991b. Perspectives on management of pestiferous Chironomidae (Diptera), an emerging global problem. Journal of the American Mosquito Control Association 7: 260-281. Ali, A., R. A. Baggs, and J. P. Stewart. 1981. Susceptibility of some Florida chironomid midges and mosquitoes to various formulations of Bacillus thuringiensis serovar. israelenisis. Journal of Economic Entomology 74: 672. Ali, A., L. C. Barbato, G. Ceretti, S. Della Sala, R. Riso, G. Marchese, and F. D'Andrea. 1992. Efficacy of two temephos formulations against Chironomus salinarius (Diptera: Chironomidae) in the saltwater Lagoon of Venice, Italy. Journal of the American Mosquito Control Association 8: 353-356. Ali, A., M. S. Mulla, A. R. Pfuntner, and L. L. Luna. 1978. Pestiferous midges and their control in a shallow residential-recreational lake in southern California. Mosquito News 38: 528 - 535. Ali, A. and J. K. Nayar. 1986. Efficacy of Bacillus sphaericus Neide against larval mosquitoes (Diptera: Culicidae) and midges (Diptera: Chironomidae) in the laboratory. Florida Entomologist 69: 685- 690. Anderson, R. L. 1989. Toxicity of synthetic pyrethroids to freshwater invertebrates. Environmental Toxicology and Chemistry 8: 403-410. Ankley, G. T., J. E. Tietge, D. L. DeFoe, K. M. Jensen, G. W. Holcombe, E. J. Durhan, and S. A. Diamond. 1998. Effects of ultraviolet light and methoprene on survival and development of Rana pipiens. Environmental Toxicology and Chemistry 17: 2530-2542. Batzer, D. P., M. McGee, V. H. Resh, and R. R. Smith. 1993. Characteristics of invertebrates consumed by mallards and prey response to wetland flooding schedules. Wetlands 13: 41- 49.

Boisvert, M. and J. Boisvert. 2000. Effects of Bacillus thuringiensis var. israelensis on target and nontarget organisms: a review of laboratory and field experiments. Biocontrol Science and Technology 10: 517-561. Breaud, T. P., J. E. Farlow, C. D. Steelman, and P. E. Schilling. 1977. Effects of the insect growth regulator methoprene on natural populations of aquatic organisms in Louisiana intermediate marsh habitats. Mosquito News 37: 704-712. Brown, M. D., D. Thomas, K. Watson, J. G. Greenwood, and B. H. Kay. 1996. Acute toxicity of selected pesticides to the estuarine shrimp Leander tenuicornis (Decapoda: Palaemonidae). Journal of the American Mosquito Control Association 12: 721-724. Calhoun, J. V., J. R. Slotten, and M. H. Salvato. 2000. The rise and fall of tropical blues in Florida: Cyclargus ammon and Cyclargus thomasi bethunebakeri (Lepidoptera: Lycaenidae). Holarctic Lepidoptera 77: 13-20. Charbonneau, C. S., R. D. Drobney, and C. F. Rabeni. 1994. Effects of Bacillus thuringiensis var. israelensis on nontarget benthic organisms in a lentic habitat and factors affecting the efficacy of the larvicide. Environmental Toxicology and Chemistry 13: 267-279.

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Cooper, C. B. and S. H. Anderson. 1996. Significance of invertebrate abundance to dabbling duck brood use of created wetlands. Wetlands 16: 557-563. Cox, R. J., M. A. Hanson, C. C. Roy, N. J. Euliss, D. H. Johnson, and M. G. Butler. 1998. Mallard duckling growth and survival in relation to aquatic invertebrates. Journal of Wildlife Management 62: 124-133. Degitz, S. J., E. J. Durhan, J. E. Tietge, P. A. Kosian, G. W. Holcombe, and G. T. Ankley. 2003. Developmental toxicity of methoprene and several degradation products in Xenopus laevis. Aquatic Toxicology 64: 97-105. Fales, J. H., P. J. Spangler, O. F. Bodenstein, G. D. Mills, Jr., and C. G. Durbin. 1968. Laboratory and field evaluation of Abate against a backswimmer (Notonecta undulata Say) (Hemiptera: Notonectidae). Mosquito News 28: 77-81. Fortin, C., A. Maire, and R. Leclair. 1987. The residual effect of temephos (Abate 4-E) on nontarget communities. Journal of the American Mosquito Control Association 3: 282- 288. Foster, W. A. 1995. Mosquito sugar feeding and reproductive energetics. Annual Review of Entomology 40: 443-474. Fox, I. 1998. Predation on Aedes aegypti (Diptera: Culicidae) by Theridion rufipes (Araneae: Theridiidae) in Puerto Rico. Journal of Medical Entomology 35: 611-613. Hagler, J. R., G. D. Waller, and B. E. Lewis. 1989. Mortality of honeybees (Hymenoptera: Apidae) exposed to permethrin and combinations of permethrin with piperonyl butoxide. Journal of Apicultural Research 28: 208-211. Helgen, J. C. 1989. Larval mosquitoes as vulnerable prey: Chaoborus predation. Canadian Journal of Fisheries and Aquatic Sciences 46: 1642-1650. Helgen, J. C., N. J. Larson, and R. L. Anderson. 1988. Response of zooplankton and Chaoborus to temephos in a natural pond and in the laboratory. Archives of Environmental Contamination and Toxicology 17: 459-471. Hershey, A. E., A. R. Lima, G. J. Niemi, and R. R. Regal. 1998. Effects of Bacillus thuringiensis israelensis (Bti) and methoprene on nontarget macroinvertebrates in Minnesota wetlands. Ecological Applications 8: 41-60. Hershey, A. E., L. Shannon, R. Axler, C. Ernst, and P. Mickelson. 1995. Effects of methoprene and Bti (Bacillus thuringiensis var. israelensis) on non-target insects. Hydrobiologia 308: 219-227. Higgins, M. J. and R. W. Merritt 1999. Temporary woodland ponds in Michigan: Invertebrate seasonal patterns and trophic relationships, p. 279-297. In D. P. Batzer, R. B. Rader, and S. A. Wissinger [eds.], Invertebrates in Freshwater Wetlands of North America: Ecology and Management. John Wiley and Sons. New York. Hill, I. R., J. L. Shaw, and S. J. Maund.Hill, I. R., Heimbach, F., Leeuwangh, P., and Mattiessen, P. [eds.] 1994. Review of Aquatic Field Tests With Pyrethroid Insecticides. Lewis Publishers. Boca Raton, FL (USA). Iannacone, J. A. and L. Alvarino. 1998. Acute ecotoxicity of the organophosphate insecticide temephos to Chironomus calligraphus Goeldi (Diptera: Chironomidae) Ecotoxicidad aguda del insecticida organofosforado tempephos sobre Chironomus calligraphus Goeldi (Diptera: Chironomidae). Acta Entomologica Chilena 22: 53-55. Jensen, T., S. P. Lawler, and D. A. Dritz. 1999. Effects of ultra-low volume pyrethrin, malathion, and permethrin on nontarget invertebrates, sentinel mosquitoes, and mosquitofish in

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seasonally impounded wetlands. Journal of the American Mosquito Control Association 15: 330- 338. Kevan, P. G. 1972. Insect pollination of high arctic flowers. Journal of Ecology 60: 831-847. Kurta, A. and J. J. Whitaker. 1998. Diet of the Endangered Indiana Bat (Myotis sodalis) on the Northern Edge of Its Range. American Midland Naturalist 140: 280-286. La Clair, J. J., J. A. Bantle, and J. Dumont. 1998. Photoproducts and metabolites of a common insect growth regulator produce developmental deformities in Xenopus. Environmental Science & Technology 32: 1453-1461. Lacey, L. A. and M. S. Mulla 1990. Safety of Bacillus thuringiensis ssp. israelensis and Bacillus sphaericus to nontarget organisms in the aquatic environment, p. 169-188. In M. Laird, L. A. Lacey, and E. Davidson [eds.], Safety of Microbial Insecticides. CRC Press. Baco Raton, FL. Lawler, S. P., D. Dritz, T. Strange, and M. Holyoak. 1999a. Effects of introduced mosquitofish and bullfrogs on the threatened California red-legged frog. Conservation Biology 13: 613- 622. Lawler, S. P., T. Jensen, D. A. Dritz, and G. Wichterman. 1999b. Field efficacy and nontarget effects of the mosquito larvicides temephos, methoprene, and Bacillus thuringiensis var. israelensis in Florida mangrove swamps. Journal of the American Mosquito Control Association 15: 446-452. Lawler, S. P., K. Miles, D. Dritz, and S. E. Spring. 1998. Effects of Golden Bear oil on nontarget aquatic organisms inhabiting salt marshes. Mosquito Control Research, Annual Report 66-71. University of California, Division of Agriculture and Natural Resources, Oakland, CA. McKenney, C. L. and D. M. Celestial. 1996. Modified survival, growth and reproduction in an estuarine mysid (Mysidopsis bahia) exposed to a juvenile hormone analogue through a complete life cycle. Aquatic Toxicology 35: 11-20. Merritt, R. W. and K. W. Cummins. 1996. An Introduction to the Aquatic Insects of North America, 3rd. ed. Kendall/Hunt. Dubuque, IA. Merritt, R. W., R. H. Dadd, and E. D. Walker. 1992. Feeding behavior, natural food, and nutritional relationships of larval mosquitoes. Annual Review of Entomology 37: 349- 376. Milam, C. D., J. L. Farris, and J. D. Wilhide. 2000. Evaluating Mosquito Control Pesticides for Effect on Target and Nontarget Organisms. Archives of Environmental Contamination and Toxicology 39: 324-328. Miura, T. and R. M. Takahashi. 1973. Insect developmental inhibitors. 3. Effects on nontarget organisms. Journal of Economic Entomology 66: 915-922. Miura, T., R. M. Takahashi, and F. I. Mulligan. 1980. Effects of the mosquito larvicide Bacillus thuringiensis serotype H-14 on selected aquatic organisms. Mosquito News 40: 619-622. Mortimer, M. R. and H. F. Chapman. 1995. Acute toxic effects of (S)-methoprene and temephos to some Australian non-target aquatic crustacean species. Australasian Journal of Ecotoxicology 1: 107- 111.

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Mulla, M. S. and H. A. Darwazeh. 1981. Efficacy of petroleum larvicidal oils and their impact on some aquatic nontarget organisms. Proceedings of the California Mosquito Control Association 49:84- 87. Mulla, M. S., H. A. Darwazeh, E. W. Davidson, H. T. Dulmage, and S. Singer. 1984. Larvicidal activity and field efficacy of Bacillus sphaericus strains against mosquito larvae and their safety to nontarget organisms. Mosquito News 44: 336-342. Mulla, M. S. and A. M. Khasawinah. 1969. Laboratory and field evaluation of larvicides against chironomid midges. Journal of Economic Entomology 62: 37-41. Mulla, M. S., R. L. Norland, T. Ikeshoji, and W. L. Kramer. 1974. Insect growth regulators for the control of aquatic midges. Journal of Economic Entomology 67: 165-170. Nayar, J. K., J. W. Knight, A. Ali, D. B. Carlson, and D. O'Bryan. 1999. Laboratory evaluation of biotic and abiotic factors that may influence larvicidal activity of Bacillus thuringiensis serovar. israelensis against two Florida mosquito species. Journal of the American Mosquito Control Association 15: 32-42. Niemi, G. J., A. E. Hershey, L. Shannon, J. M. Hanowski, A. Lima, R. P. Axler, and R. R. Regal. 1999. Ecological effects of mosquito control on zooplankton, insects, and birds. Environmental Toxicology and Chemistry 18: 549-559. Nilsson, A. N. and O. Soederstroem. 1988. Larval consumption rates, interspecific predation, and local guild composition of egg-overwintering Agabus (Coleoptera, Dystiscidae) species in vernal ponds. Oecologia 76: 131-137. Norland, R. L. and M. S. Mulla. 1975. Impact of Altosid on selected members of an aquatic ecosystem. Environmental Entomology 4: 145-152. Olmstead, A. W. and G. L. LeBlanc. 2001. Low exposure concentration effects of methoprene on endocrine-regulated processes in the crustacean Daphnia magna. Toxicological Sciences 62: 268- 273. Pankiw, T. and S. C. Jay. 1992a. Aerially applied ultra-low-volume malathion effects on caged honey bees (Hymenoptera: Apidae), caged mosquitoes (Diptera: Culicidae), and malathion residues. Journal of Economic Entomology 85: 687-691. Pankiw, T. and S. C. Jay. 1992b. Aerially applied ultra-low-volume malathion effects on colonies of honey bees (Hymenoptera: Apidae). Journal of Economic Entomology 85: 692-699. Pierce, R., M. Henry, D. Kelly, P. Sherblom, W. Kozlowsky, G. Wichterman, and T. W. Miller. 1996. Temephos distribution and persistence in a southwest Florida salt marsh community. Journal of the American Mosquito Control Association 12: 637-646. Pinkney, A. E., P. C. McGowan, D. R. Murphy, T. P. Lowe, D. W. Sparling, and L. C. Ferrington. 2000. Effects of the mosquito larvicides temephos and methoprene on insect populations in experimental ponds. Environmental Toxicology and Chemistry 19: 678- 684. Pinkney, A. E., P. C. McGowan, D. R. Murphy, T. P. Lowe, D. W. Sparling, and W. H. Meredith. 1999. Effects of temephos (Abate registered 4E) on fiddler crabs (Uca pugnax and Uca minax) on a Delaware salt marsh. Journal of the American Mosquito Control Association 15: 321-329. Pont, D., E. Franquet, and J. N. Tourenq. 1999. Impact of different Bacillus thuringiensis variety israelensis treatments on a chironomid (Diptera Chironomidae) community in a temporary marsh. Journal of Economic Entomology 92: 266-272.

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Robert, N. and P. Venkatesan. 1997. Prey preference and predatory efficiency of the water bug, Diplonychus indicus Venk. & Rao (Hemiptera: Belostomatidae), an effective biocontrol agent for mosquitoes. Journal of Entomological Research 21: 267-272. Rodcharoen, J., M. S. Mulla, and J. D. Chaney. 1991. Microbial larvicides for the control of nuisance aquatic midges (Diptera: Chironomidae) inhabiting mesocosms and man-made lakes in California. Journal of the American Mosquito Control Association 7: 56-62. Ross, D. H., D. Judy, B. Jacobson, and R. Howell. 1994. Methoprene concentrations in freshwater microcosms treated with sustained-release Altosid formulations. Journal of the American Mosquito Control Association 10: 202-210. Rupp, H. R. 1996. Adverse assessments of Gambusia affinis: An alternate view for mosquito control practitioners. Journal of the American Mosquito Control Association 12: 155-166. Safurabi, S. and J. I. Madani. 1999. Prey preference of an aquatic beetle Dineutes indicus Aube (Coleoptera: Gyrinidae). Journal of Ecobiology 11: 237-240. Salvato, M. H. 2001. Influence of mosquito control chemicals on butterflies (Nymphalidae, Lycaenidae, Hesperiidae) of the lower Florida Keys. Journal of the Lepidopterists' Society 55: 8-14. Service, M. W. 1993. Mosquito Ecology: Field Sampling Methods, 2nd ed. Chapman and Hall. London. Siegfried, B. D. 1993. Comparative toxicity of pyrethroid insecticides to terrestrial and aquatic insects. Environmental Toxicology and Chemistry 12: 1683-1689. Sparling, D. W., T. P. Lowe, and A. E. Pinkney. 1997. Toxicity of Abate registered to green frog tadpoles. Bulletin of Environmental Contamination and Toxicology 58: 475-481. Spielman, A. and M. D'Antonio. 2001. Mosquito: The Story of Man's Deadliest Foe. Hyperion. New York. Steelman, C. D., J. E. Farlow, and T. P. Breaud. 1975. Effects of growth regulators on Psorophora columbiae (Dyar and Knab) and non-target aquatic insect species in rice fields. Mosquito News 35: 67-76. Strickman, D., R. Sithiprasasna, and D. Southard. 1997. Bionomics of the spider, Crossopriza lyoni (Araneae, Pholcidae), a predator of dengue vectors in Thailand. Journal of Arachnology 25: 194-201. Sukhacheva, G. A. 1996. Study of the natural diet of adult dragonflies using an immunological method. Odonatologica 25: 397-403. Taylor, K. S., G. D. Waller, and L. A. Crowder. 1987. Impairment of a classical conditioned response of the honey bee (Apis mellifera L.) by sublethal doses of synthetic pyrethroid insecticides. Apidologie 18: 243-252. Tyler, C. R., N. Beresford, M. van der Woning, J. P. Sumpter, and K. Thorpe. 2000. Metabolism and degradation of pyrethroid insecticides produce compounds with endocrine activities. Environmental Toxicity and Chemistry 19: 801-809.

Urabe, K., T. Ikemoto, and S. Takei. 1990. Studies on Sympetrum frequens (Odonata: Libellulidae) nymphs as natural enemies of the mosquito larvae, Anopheles sinensis , in rice fields. 4. Prey- predator relationship in the rice field areas. Japanese Journal of Sanitary Zoology 41: 265-272. K-20

USEPA. 1999. Revised Environmental Fate and Effects Division Reregistration Eligibility Document for Temephos. U.S. Environmental Protection Agency, Washington, D.C. USEPA. 2000. Malathion Reregistration Eligibility Document, Environmental Fate and Effect Chapter (Revised). U.S. Environmental Protection Agency, Washington, D.C. Ward, D. V. and D. A. Busch. 1976. Effects of temephos, an organophosphate insecticide, on survival and escape behavior of the marsh fiddler crab Uca pugnax. Oikos 27: 331-335. Yiallouros, M., V. Storch, and N. Becker. 1999. Impact of Bacillus thuringiensis var. israelensis on Larvae of Chironomus thummi thummi and Psectrocladius psilopterus (Diptera: Chironomidae). Journal of Invertebrate Pathology 74: 39-47. Yousten, A. A., F. J. Genthner, and E. F. Benfield. 1992. Fate of Bacillus sphaericus and Bacillus thuringiensis serovar israelensis in the aquatic environment. Journal of the American Mosquito Control Association 8: 143-148. Zinn, T. L. and S. R. Humphrey. 1981. Seasonal food resources and prey selection of the southeastern brown bat (Myotis austroriparius ) in Florida. Florida Scientist 44: 81-90.

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APPENDIX J. Mosquito Biology

Mosquito Biology (USFWS 2004)

The following species descriptions provide information about some important mosquito vectors of human arboviruses in the U.S. An arbovirus is any virus that is maintained and biologically transmitted through vertebrate hosts by blood-feeding arthropod vectors (e.g., ticks and mosquitoes). The virus multiplies within a competent vector and is passed to a new host through subsequent blood meals. Examples of arboviruses include various forms of encephalomyelitis as well as dengue. Note that this section is not all- inclusive there are more species of mosquitoes that represent potential arbovirus vectors. The following species represent primary and important bridge vectors.

Aedes aegypti

Mosquitoes in the genera Aedes and Aedes have eggs that are capable of withstanding dry and, often, freezing conditions. For this reason, eggs deposited in a dry habitat may remain viable for years before hatching. Females deposit eggs on moist surfaces that will flood at a later time. Species of Aedes and Aedes mosquitoes inhabit a very diverse range of ephemeral habitats, ranging from small containers to large wetlands.

Larval Habitat/Ecology: Aedes aegypti is a tropical/sub-tropical species of mosquito introduced into the United States. Because it does not tolerate cold weather, it is found exclusively in the Southeast, especially in Florida. Aedes aegypti larvae occur exclusively in artificial container habitats and natural treeholes. Their preference for containers makes this species a serious pest in urban areas.

Number of generations per year: several

Flight range: one mile or less

Blood-feeding Host: Aedes aegypti feeds primarily on mammals (including humans).

Disease Association: Aedes aegypti is a primary vector of the human arboviruses Dengue and Yellow Fever throughout the tropical and sub-tropical parts of the world. It has been found with WN in 2002, but it preference for mammalian blood probably makes it relatively unimportant as a bridge vector for zoonotic arboviruses.

Aedes albopictus (Asian tiger mosquito)

Larval Habitat/Ecology: Aedes albopictus is an exotic species of mosquito introduced into the United States during the early 1980s via used tire shipments from Asia. Since its introduction, it has spread widely over the southern U.S. and along the Mid-Atlantic coast. Aedes albopictus larvae occur primarily in container habitats, either natural or artificial. While they are frequently associated with discarded tires, they may also be found in any container that will hold water, including natural treeholes. Females oviposit on the sides of containers, and the eggs hatch when the container is flooded. Aedes albopictus is often the most serious mosquito pest in urban and

residential areas within its range. It is rarely abundant in natural areas.

Number of generations per year: several

Flight range: one mile or less (much of the rapid dispersal of this species throughout the southern and eastern U.S. is the result of human activities such as movements of used tires).

Blood-feeding Host: Aedes albopictus feeds primarily on mammals (including humans), but will occasionally take blood meals from birds. Females are aggressive and will readily bite during the daylight hours as well as at dusk.

Disease Association: Aedes albopictus is a competent experimental vector of several arboviruses, including Dengue, Eastern Equine Encephalitis (EEE), West Nile virus (WN), and Cache Valley. Although EEE virus has been isolated from field-collected Aedes albopictus, its role in the transmission of this disease remains uncertain. West Nile Virus has been detected in this species on numerous occasions, suggesting that it is an important bridge vector of this disease to humans.

Aedes vexans (vexans mosquito, inland floodwater mosquito)

Larval Habitat/Ecology: Larvae of Aedes vexans may be found in small snowmelt pools in the spring, but are much more common in temporary rain-filled pools from late spring through late summer. Principal habitats include (freshwater) shallow, grass-filled depressions, temporary woodland pools, and shallow pools formed from irrigation. Larvae in a suitable habitat can reach extremely high population numbers, with an estimate of 80 million/ha recorded at one site in Canada. Because hatching usually occurs in the summer when water temperatures are warm, larval development can be rapid, from egg to biting adult in less than a week. Females oviposit in dry basins where the eggs will remain viable until the habitat is flooded.

Number of generations per Year: One to several.

Flight Range: up to 5-10 miles

Blood-feeding Host: Mammals are preferred, but Aedes vexans will occasionally feed on birds, making this species an important bridge vector for encephalitis viruses. Aedes vexans feeds primarily at dusk, but will also bite during the day in shaded areas.

Disease Association: Aedes vexans has been implicated (as a bridge vector) in the disease cycle of Eastern Equine Encephalitis (EEE), Western Equine Encephalitis (WEE), and West Nile virus (WN).

Aedes canadensis (woodland pool mosquito)

Larval Habitat/Ecology: Larvae occur in small snowmelt pools in woodland habitats. They hatch slightly later than other species of Aedes found in these same habitats, and may also hatch after summer rains flood these woodland basins. Adults generally emerge in May, and may be encountered at any time from late spring through late summer. Adults usually remain near their

woodland larval habitats and are apparently long-lived.

Number of Generations per Year: one or two

Flight Range: usually one mile or less

Blood-feeding Host: Aedes canadensis prefers mammals (including humans) but will readily feed on birds as well. They bite predominantly during the day.

Disease Association: Aedes canadensis has been implicated in the disease cycle of EEE and California Encephalitis (CE). Because it will feed on both birds and mammals (including humans), and because adults tend to be long-lived, it is a potentially important bridge vector. More research is needed to assess the vector potential of this species.

Aedes dorsalis

Larval Habitat/Ecology: Aedes dorsalis is found throughout the U.S. except for the Southeast, but is most common in the West. Larvae occur in shallow, open floodwaters with grassy vegetation. The larvae can be found in both saline and alkaline freshwater habitats, often in very large numbers. Along the Pacific Coast it is common in tidal marshes.

Number of Generations per Year: Several

Flight Range: several miles (up to 20-30)

Blood-feeding Host: This species feeds almost exclusively on mammals, usually in the evening hours. Females of this species are aggressive biters.

Disease Association: Aedes dorsalis is apparently important as a vector of California Encephalitis, in which the disease cycle is in small mammals. The virus can be passed directly into the mosquito=s eggs, so emerging adults can already be infected with the disease.

Aedes melanimon

Larval Habitat/Ecology: The larval habitats of Aedes melanimon are irrigated pastures and shallow, open flooded areas in the West. Larvae of this species occur only in freshwater and are often associated with wetlands flooded for waterfowl habitat. Like many other species of floodwater mosquitoes, Aedes melanimon densities can be very high in recently flooded habitats.

Number of Generations per Year: Several.

Blood-feeding Host: Aedes melanimon feeds primarily on mammals, with hares and rabbits favored hosts. This species will also readily bite humans. Blood-feeding generally takes place in the evening and early morning hours.

Flight Range: up to 10 miles

Disease Association: Aedes melanimon has been implicated as a vector of Western Equine Encephalitis (in a cycle involving hares and rabbits) and in California Encephalitis.

Aedes washinoi

Larval Habitat/Ecology: The larval habitats of Aedes washinoi are snowmelt pools and floodwater habitats of valleys and foothills to about 1300 m above sea level. Until recently, this species was grouped with A. increpitus. Occurs primarily in California.

Number of Generations per Year: 2 or more at lower elevations.

Blood-feeding Host: Aedes washinoi feeds primarily on mammals, and will readily bite humans. Females feed readily during the day.

Flight Range: less than one mile

Disease Association: Unknown

Coquilletidia ( = Mansonia) perturbans (irritating mosquito)

Larval Habitat/Ecology: Coquilletidia perturbans is a resident of permanent freshwater marshes that have soft muck bottoms and abundant emergent vegetation. The larvae are unusual for mosquitoes in that they remain buried in the soft sediments and obtain oxygen by piercing the roots of aquatic plants with a specialized siphon. They can be found on a variety of aquatic plants including Typha, Sagittaria, Pontederia, Nymphaea, Juncus, and Carex. Larvae overwinter attached to roots and emerge as adults in June and July. Because of their unique means of obtaining oxygen, larvae of Coquilletidia perturbans are not as mobile as other mosquito larvae and are especially susceptible to desiccation from declining water levels (a factor used in control of this species). Adult females oviposit directly on the surface of the water in areas of emergent vegetation. The eggs hatch after a short time and the larvae attach themselves to the roots of plants in the muck, where they remain throughout the winter.

Number of Generations per Year: One (two in Florida).

Flight Range: several miles

Blood-feeding Host: Coquilletidia perturbans is somewhat of a generalist feeder, apparently preferring mammals but readily taking blood meals from birds as well. They bite predominantly at dusk.

Disease Association: Coquilletidia perturbans is a primary vector of EEE. Because it readily feeds on both birds and mammals, this species is believed to be important in transmitting EEE virus to horses and humans. It may also be an important bridge vector of WN.

Culex nigripalpus

Mosquitoes in the genus Culex generally breed in water containing a high amount of organic material. Eggs are deposited directly on the surface of the water (unlike eggs of Aedes/Aedes, they are not desiccation-resistant) and all species are capable of multiple generations per year. All species of Culex overwinter as adults.

Larval Habitat/Ecology: Culex nigripalpus is found in freshwater ditches and marshes in the Southeast, especially Florida. Like other species in this genus, females lay eggs directly on the water surface, and breeding is continuous throughout the warmer months of the year.

Number of Generations per Year: several

Flight Range: several miles

Blood-feeding Host: Culex nigripalpus is a generalist feeder, taking blood meals from both birds and mammals. They bite predominantly after dusk.

Disease Association: Culex nigripalpus is a competent vector of SLE and WN. It readily feeds on both birds and mammals, making it a potentially important bridge vector, and may be capable of maintaining the virus cycle in birds.

Culex pipiens and Culex pipiens quinquefasciatus (northern and southern house mosquito)

Larval Habitat/Ecology: As the common names imply, Culex pipiens and Culex p. quinquefasciatus (two closely related species/subspecies) are usually associated with human habitations. Larvae occur in water with a high organic content, and are often found in artificial container habitats such as bird baths, children=s wading pools, storm sewers, rain gutters, tires, and even tin cans. Because of their preference for these habitats, Culex pipiens and Culex p. quinquefasciatus are often very abundant in urban centers, where artificial containers are often numerous and widespread. They are also common in small ground pools in both urban and natural areas. Adult females oviposit directly on the surface of the water and eggs hatch shortly thereafter. In small habitats during the summer, larval development can be rapid. Thus, although water is required for egg laying and larval development, even containers that hold water for only a week or so can provide suitable breeding habitat. Adults overwinter in buildings, sewers, etc.

Number of Generations per Year: Several.

Flight Range: at least 1 mile.

Blood-feeding Host: Both Culex pipiens and Culex p. quinquefasciatus appear to prefer birds to mammals, especially in the late spring and early summer. Later in the summer, however, feeding on mammals (including humans) seems to increase, especially for Culex p. quinquefasciatus. Adults will readily enter houses to feed on humans. They bite primarily from dusk to midnight.

Disease Association: Culex pipiens and Culex p. quinquefasciatus are primary vectors for St. Louis Encephalitis (SLE). Culex pipiens and Culex p. quinquefasciatus have also been identified as primary vectors of West Nile virus (WN) in the East and South, respectively. Both SLE and WN are amplified within bird populations in the spring and early summer, when the mosquitoes are feeding primarily on birds.

Culex restuans (white dotted mosquito)

Larval Habitat/Ecology: Larvae of Culex restuans occupy similar habitats as Culex pipiens and Culex p. quinquefasciatus: water with a high organic content. Ditches, open ground pools, and containers are preferred habitats. Adults are most abundant during the spring and early summer (less so in late summer), and again in the autumn. Adults overwinter in similar habitats as Culex pipiens.

Number of Generations per Year: several

Flight Range: 1-3 miles

Blood-feeding Host: Culex restuans appears to feed primarily on birds, rarely biting humans. They bite primarily at dusk.

Disease Association: Culex restuans is a competent vector of SLE virus and WN. Its preference for birds and its early season abundance suggest that it is important in amplifying virus among birds in the spring and early summer. It is probably relatively unimportant as a bridge vector.

Culex tarsalis

Larval Habitat/Ecology: This species is much more common in the southern, central, and western U.S than it is in the eastern U.S. Culex tarsalis larvae are found in irrigation ditches, ponds, and storm sewers, usually that contain abundant organic material. As in other species of Culex, overwintering is in the adult stage.

Number of Generations per Year: several

Flight Range: up to 25 miles

Blood-feeding Host: Culex tarsalis is a generalist feeder. While birds may be preferred, it will readily feed on mammals (including humans) as well. They bite primarily after dusk.

Disease Association: Culex tarsalis is the primary vector of Western Equine Encephalitis in the Midwest and West. It is also an efficient vector of SLE. West Nile virus was detected in this species in 2002, and laboratory tests indicate it is a very competent vector of WN. Vertical transmission of WN (i.e., from adult female to her eggs) has been demonstrated in the laboratory.

Culiseta inornata

Larval Habitat/Ecology: Larvae of Culiseta inornata occur in a wide variety of permanent and semi- permanent aquatic habitats, including impoundments, duck club ponds, ditches and seepages. Although primarily a freshwater species, Cs. inornata may also be found in brackish water. In the southern part of its range (including much of California) this species is most abundant in the fall, winter, and spring, with adult females aestivating during the hot, dry summers.

Number of Generations per Year: 2 or more

Flight Range: several miles

Blood-feeding Host: Culiseta inornata feeds primarily on large domestic animals, but will readily take a blood meal from a human host.

Disease Association: Culiseta inornata has been implicated in the transmission of Western Equine Encephalitis and the California Encephalitis group of viruses. It has been demonstrated in the laboratory to be capable of transmitting SLE and WN.

Psorophora sp.

Psorophora mosquitoes are generally large species that can inflict a painful bite. These mosquitoes develop in rain-filled pools during the summer, frequently with Aedes vexans. Females deposit drought- resistant eggs in dry basins in the summer and fall, and the eggs do not hatch until inundated with warm water the following summer. They are capable of multiple generations per year, especially in warmer climates. The three most common species in the U.S.

are Psorophora ciliata, Ps. columbiae, and Ps. ferox. Psorophora ciliata are unique in that late- stage larvae are predaceous, often consuming other mosquito larvae. Aside from being troublesome biters, Psorophora mosquitoes are not important vectors of endemic arborviruses in most of the U.S. However, they have been implicated in outbreaks of Venezuelan Equine Encephalitis in southern Texas and Mexico. These mosquitoes may fly one to several miles from their breeding habitat.

Anopheles sp.

Anopheles mosquitoes are common in permanent to semi-permanent bodies of water. Females overwinter as adults and deposit eggs directly on the surface of the water during the summer (eggs are not drought- resistant). Several generations per year are produced. Anopheles larvae are easily distinguished from other genera by their horizontal position at the water surface, compared with other mosquitoes that rest more-or-less vertically at the surface. Anopheles mosquitoes are generally not important vectors of arboviruses, but are the primary vectors of malaria. A common species in the East is Anopheles quadrimaculatus and in the West, Anopheles freeborni. The flight range of Anopheles mosquitoes is generally about 1-2 miles, but some may fly several miles from their larval habitat.

Table 3. Summary of Important Mosquito Vectors of Human Arboviruses in the United States

Voltinis Disease Mosquito species Range in US Larval Habitat Type3 Principal Host m Vector2

Aedes aegypti SE containers M Dengue (Den) P m (humans) containers, EEE(?), WN, Aedes albopictus S, E M B m (occas. b) treeholes Den summer rain EEE, WEE, Aedes vexans all US M B m:66-99+ pools WN Aedes canadensis E 2/3 spring pools U or M EEE, WN(?) B m saltmarsh, Aedes dorsalis US, ex. SE M CE, SLE, WEE B m brackish Aedes melanimon W grassy pools M CE, WEE B m E, SE coast, S Aedes taeniorhynchus saltmarsh M EEE(?), WN(?) B m Calif. US, ex. Far summer rain Aedes trivittaus M WN(?) B m West pools US, ex. High cattail, sedge U, M in Coquillettidia perturbans EEE, WN B m:90 Plains marsh FL ditches, FW Culex nigripalpus SE M EEE, SLE, WN P,B generalist marshes Culex pipiens N 2/3 grnd. pools, M SLE, WN P,B b (occas. m) grnd. pools, Culex quinquefasciatus S 1/2 M SLE, WN P,B b and m containers grnd. pools, Culex restuans most US M SLE, WN P b containers W; most US, irrig. ditches, WEE, SLE, WN Culex tarsalis M P,B b (spring) m (sum.) ex. NE ponds

1 U: Univoltine, one brood M: Multivoltine, 2+ broods 2 CE: California Encephalitis EEE: Eastern Equine Encephalitis LAC: LaCrosse Encephalitis SLE: St. Louis Encephalitis WEE:

Western Equine Encephalitis; WN: West Nile Virus Den: Dengue 3 b: birds; m: mammals; 4 Number is %

APPENDIX K. Pyrethrins and Pyrethroids

PYRETHRINS AND PYRETHROIDS (http://extoxnet.orst.edu/pips/pyrethri.htm)

Produced By the Extension Toxicology Network

Pesticide Information Profiles

A Pesticide Information Project of Cooperative Extension Offices of Cornell University, Oregon State University, the University of Idaho, and the University of California at Davis and the Institute for Environmental Toxicology, Michigan State University. Major support and funding was provided by the USDA/Extension Service/National Agricultural Pesticide Impact Assessment Program.

EXTOXNET primary files maintained and archived at Oregon State University Revised

3/94

PYRETHRINS AND PYRETHROIDS

TRADE OR OTHER NAMES: Several trade names associated with these compounds are Buhach, Chrysanthemum Cinerariaefolium, Ofirmotox, Insect Powder, Dalmation Insect Flowers, Firmotox, Parexan and NA 9184.

INTRODUCTION: Pyrethrins are natural insecticides produced by certain species of the chrysanthemum plant. The flowers of the plant are harvested shortly after blooming and are either dried and powdered or the oils within the flowers are extracted with solvents. The resulting pyrethrin containing dusts and extracts usually have an active ingredient content of about 30%. These active insecticidal components are collectively known as pyrethrins. Two pyrethrins are most prominent, pyrethrin-I and pyrethrin-II. The pyrethrins have another four different active ingredients, Cinerin I and II and Jasmolin I and II. The typical composition of pyrethrin is pyrethrin I (38.0%), cinerin I (7.3%), jasmolin I (4.0%), pyrethrin II (35.0%), cinerin II (11.7%) and jasmolin II (4.0%); the composition of piperonyl butoxide is 80% 5-[2-(2-butyloxyethoxy)ethoxymethyl]-6- propyl-1,3-benzodioxole and 20% related compounds. Pyrethrin compounds have been used primarily to control human lice, mosquitoes, cockroaches, beetles and flies. Some "pyrethrin dusts," used to control insects in horticultural crops, are only 0.3% to 0.5% pyrethrins, and are used at rates of up to 50 lb/A. Other pyrethrin compounds may be used in grain storage and in poultry pens and on dogs and cats to control lice and fleas. The natural pyrethrins are contact poisons which quickly penetrate the nerve system of the insect. A few minutes after application, the insect cannot move or

fly away. But, a "knockdown dose" does not mean a killing dose. The natural pyrethrins are swiftly detoxified by enzymes in the insect. Thus, some pests will recover. To delay the enzyme action so a lethal dose is assured, organophosphates, carbamates, or synergists may be added to the pyrethrins. Semisynthetic derivatives of the chrysanthemumic acids have been developed as insecticides. These are called pyrethroids and tend to be more effective than natural pyrethrins while they are less toxic to mammals. One common synthetic pyrethroid is allethrin. In this report, the term "pyrethrins" refers to the natural insecticides derived from chrysanthemum flowers; "pyrethroids" are the synthetic chemicals, and "pyrethrum" is a general name covering both compounds. The EPA classifies pyrethrin-I as a Restricted Use Pesticide (RUP). Restricted Use Pesticides may be purchased and used only by certified applicators.

TOXICOLOGICAL EFFECTS Acute Toxicity: Synthetic pyrethroid compounds vary in their toxicity as do the natural pyrethrins. Pyrethrum carries the signal word CAUTION. Inhaling high levels of pyrethrum may bring about asthmatic breathing, sneezing, nasal stuffiness, headache, nausea, incoordination, tremors, convulsions, facial flushing and swelling, and burning and itching sensations (102). The most severe poisonings have been reported in infants, who are not able to efficiently break down pyrethrum. The lowest lethal oral dose of pyrethrum is 750 mg/kg for children and 1,000 mg/kg for adults (102). Oral LD50 values of pyrethrins in rats range from 200 mg/kg to greater than 2,600 mg/kg (96). Some of this variability is due to the variety of constituents in the formulation. Mice have a pyrethrum oral LD50 of 370 mg/kg (102). Animals exposed to toxic amounts may experience tongue and lip numbness, nausea, and diarrhea. Symptoms may also include incoordination, tremors, convulsions, paralysis, respiratory failure, and death. Pyrethroids can cause two quite different responses at near lethal doses in rats; aggressive sparring and a sensitivity to external stimuli progressing to tremors is the one response and pawing and burrowing behavior, and salivation leading to chronic seizures is the other (105). Human response to these two different types of pyrethroids has not yet been evaluated. Recovery from serious poisoning in mammals is fairly rapid. Rats and rabbits are not affected by large dermal applications (96, 102). On broken skin, pyrethrum produces irritation and sensitization, which is further aggravated by sun exposure.

Chronic Toxicity: Absorption of pyrethrum through the stomach and intestines and through the skin is slow. However, humans can absorb pyrethrum more quickly through the lungs during respiration. Response appears to depend on the pyrethrum compound used. Overall, pyrethrins and pyrethroids are of low chronic toxicity to humans and the most common problems in humans have resulted from the allergenic properties of pyrethrum (104). Patch tests for allergic reaction are an important tool in determining an individuals sensitivity to these compounds. Many of the natural and synthetic compounds can produce skin irritation, itching, pricking sensations and local burning sensations. These symptoms may last for about two days (105).

Reproductive Effects: Rabbits that received pyrethrins orally at high doses during the sensitive period of pregnancy had normal litters. A group of rats fed very high levels of pyrethrins daily for three weeks before first mating had litters with weanling weights much lower than normal (96). Overall, pyrethrins appear to have low reproductive toxicity.

Teratogenic Effects: The one rabbit reproduction study performed showed no effect of pyrethrins on development of the offspring (101). More information is needed.

Mutagenic Effects: No information was found.

Carcinogenic Effects: No carcinogenic status has been established for pyrethrins or pyrethroids.

Organ Toxicity: In mammals, tissue storage has not been recorded. At high doses, pyrethrum can be damaging to the central nervous system and the immune system. When the immune system is attacked by pyrethrum, allergies can be worsened. Animals fed large doses of pyrethrins may experience liver damage.

Rats fed pyrethrin at high levels for two years showed no significant effect on survival, but slight, definite damage to the livers was observed (96). Inhalation of high doses of pyrethrum for 30 minutes each day for 31 days caused slight lung irritation in rats and dogs (102). Fate in Humans and Animals: Pyrethrins, pyrethroids, and their metabolites are not known to be stored in the body nor excreted in the milk (100). The urine and feces of people given oral doses of pyrethrum contain chrysanthemumic acid and other metabolites (100, 96). These metabolites are less toxic to mammals than are the parent compounds (101). Pyrethrins I and II are excreted unchanged in the feces (100). Other pyrethrum components undergo rapid destruction and detoxification in the liver and gastrointestinal tract (96).

ECOLOGICAL EFFECTS Pyrethrin is extremely toxic to aquatic life, such as bluegill and lake trout while it is slightly toxic to bird species, such as mallards. Toxicity increases with higher water temperatures and acidity. Natural pyrethrins are highly fat soluble, but are easily degraded and thus do not accumulate in the body. These compounds are toxic to bees also.Because pyrethrin-I, pyrethrin-II, and allethrin have multiple sites in their structures that can be readily attacked in biological systems, it is unlikely that they will concentrate in the food chain (100).

ENVIRONMENTAL FATE Two pyrethroid synthetic insecticides, permethrin and cypermethrin, break down in plants to produce a variety of products (103). Pyrethrins have little residual effect. In stored grain, 50% or more of the applied pyrethrins disappear during the first three or four months of storage. At least 80% of what remains is removed by handling, processing, and cooking (101). Pyrethrins alone provide limited crop protection because they are not stable. As a result, they are often combined with small amounts of antioxidants to prolong their effectiveness. Pyrethrum compounds are broken down in water to nontoxic products. Pyrethrins are inactivated and decomposed by exposure to light and air. Pyrethrins are also rapidly decomposed by mild acids and alkalis. Stored pyrethrin powders lose about 20% of their potency in one year. As the pyrethrins are purified, their stability decreases; thus, pure pyrethrin-I and pyrethrin-II are the least stable of the pyrethrins (96). Purified pyrethrins are very expensive and are only available for laboratory uses.

PHYSICAL PROPERTIES AND GUIDELINES Physical Properties: Appearance: The pyrethrins are viscous brown resins, liquids, or solids which inactivate readily in air. Chemical Name: n/a

CAS Number: 8003347 Molecular Weight: Due to differences in the types and amounts of esters in the pyrethrum mixture, its molecular weight ranges from 316 to 374. Water Solubility: considered to be insoluble in water. Solubility in Other Solvents: soluble in organic solvents like: alcohol, kerosene, nitromethane, petroleum ether, carbon tetrachloride, and ethylene dichloride. Melting Point: n/a Vapor Pressure: about 0 mm/Hg Partition Coefficient: n/a Adsorption Coefficient: n/a Exposure Guidelines: ADI: 0.04 mg/kg body weight (humans) (101) MCL: Not Available RfD: Not Available PEL: 5 mg/m3 HA: Not Available TLV: 5 mg/m3 BASIC MANUFACTURER There are several manufacturers of products in this category. REFERENCES References for the information in this PIP can be found on the website, http://extoxnet.orst.edu/pips/pyrethri.htm) (Reference List Number 2)

DISCLAIMER: The information in this profile does not in any way replace or supersede the information on the pesticide product label/ing or other regulatory requirements. Please refer to the pesticide product label/ing.

APPENDIX l. Summary of Connecticut Mosquito Trapping and Testing Results - 2016

Mosquito Trapping and Testing Cumulative Results Week of October 24, 2016

Updated October 27

The following numbers of mosquitoes were collected, identified, and tested at The Connecticut Agricultural Experiment Station in New Haven. The towns and locations of the sites, number of mosquitoes tested, and virus isolations are listed. The total number tested to date: 173,988

WNV = West Nile virus EEE = Eastern Equine Encephalitis virus JC = Jamestown Canyon virus

The closest sites to the Great Meadows Unit at Stewart B. McKinney National Wildlife Refuge are highlighted in YELLOW.

Pos. Number of Mosquito Species Town Trap Site or WNV, EEE, JC Mosquitoes (No.) Neg.

Barkhamsted Hoyt Hayes Swamp 508 Neg. - -

Bethany Bethany Bog 811 Neg. - -

An. Bethel Meckauer Park 289 Pos. JC(1) quadrimaculatus (1)

Branford Hosley Ave. 1,089 Neg. - -

Bridgeport Beardsley Zoo 2,180 Pos. WNV(6) Cx. pipiens (6)

Bridgeport Barnum Blvd. 3,676 Neg. - -

Cx. pipiens (2) Bridgeport Central High School 2,773 Pos. WNV(3) Ae. albopictus (1)

Canaan Robin’s Swamp 1,976 Neg. - -

Cheshire Lock 12 566 Neg. - -

CockaponsetSt. Oc. abserratus (1) Chester 1,338 Pos. JC(2) Forest Oc. canadensis (1)

Cornwall Mohawk Pond 1,768 Neg. - -

Cromwell Cromwell Meadows 6,457 Pos. WNV(1) An. walkeri (1)

Danbury Reservoir Road 2,356 Neg. - -

Darien Brush Island Road 13,357 Pos. WNV(4) Cx. pipiens (4)

Darien High School Lane 1,557 Pos. WNV(3) Cx. pipiens (3)

Pos. WNV(1) Cx. pipiens (1) East Haven Kenneth Street 6,425 Pos. JC(4) Oc. aurifer (4)

Easton Sport Hill Road 821 Pos. WNV(1) Cs. morsitans (1)

Fairfield Catamount Road 1,293 Pos. JC(1) Oc. canadensis (1)

Cx. pipiens (5) Fairfield Pine Creek Area 657 Pos. WNV(6) Cx. restuans (1)

Farmington Shade Swamp 1,371 Neg. - -

Franklin Wildlife Refuge 2,865 Neg. - -

Glastonbury Tryon Street 1,474 Neg. - -

Greenwich Civic Center 1,314 Neg. - -

Greenwich Lake Avenue 322 Neg. - -

Greenwich Mianus River Park 562 Neg. - -

Groton U. S. Naval Base 2,908 Neg. - -

Guilford Moose Hill Road 2,233 Neg. - -

Haddam Little City Road 1,447 Neg. - -

Hamden Lake Wintergreen 952 Neg. - -

Hampton Hampton Reservoir 2,964 Neg. - -

Cx. pipiens (3) Cx. restuans(1) Hartford Keney Park 1,550 Pos. WNV(6) An. quadrimaculatus(1) Oc. japonicus(1)

Raymond Brook Hebron 2,239 Neg. - - Marsh

Killingworth Chittenden Road 532 Neg. - -

Ledyard Cedar Swamp 2,725 Neg. - -

Litchfield/Morris White Memorial 1,671 Neg. - -

Lyme Cedar Lake 702 Neg. - -

Cedar Swamp, Rte. Madison 1,292 Neg. - - 80

Manchester Oak Grove Street 970 Pos. WNV(2) Cx. pipiens(2)

Meriden Falcon Park 922 Neg. - -

Middlefield Durham Meadows 4,328 Neg. - -

Milford Erna Avenue 1,679 Pos. WNV(4) Cx. pipiens(4)

Milford Fresh Meadow Lane 1,115 Pos. WNV(1) Cx. pipiens(1)

Monroe Garder Road 1,188 Neg. - -

New Britain Village Square Drive 7,922 Pos. WNV(3) Cx. pipiens(3)

New Canaan Hoyts Swamp 1,034 Neg. - -

New Canaan Michigan Road 325 Neg. - -

New Haven Beaver Pond Park 2,455 Pos. WNV(6) Cx. pipiens(6)

Cx. pipiens (1) Newington Churchill Park 1,925 Pos. WNV(2) An. punctipennis(1)

Newtown Key Rock Road 774 Neg. - -

North Branford Cedar Pond 747 Neg. - -

Quinnipiac River North Haven 782 Neg. - - Park

North Pawcatuck River 806 Neg. - - Stonington

North Bell Cedar Swamp 842 Neg. - - Stonington

North Exit 93 634 Pos. JC(1) Oc. stimulans (1) Stonington

North Wyassup Lake 2,036 Neg. - - Stonington

Norwalk Cranbury Park 612 Neg. - -

Norwalk Rowayton School 1,249 Neg. - -

Old Lyme Great Island 936 Neg. - -

Oc. abserratus (2) Orange Meeting House Lane 2,103 Pos. JC(4) Oc. aurifer (2)

An. Plainfield Cedar Swamp 1,389 Pos. JC(1) punctipennis (1)

Redding Lyons Swamp 382 Neg. - -

Ridgefield Great Swamp 750 Pos. JC(1) Oc. aurifer (1)

Shelton Shelton Avenue 1,793 Neg. - -

South Windsor Burgess Road 3,128 Pos. JC(1) Oc. excrucians (1)

East Rd/Kensington Southington 358 Neg. - - Rd

Stafford Cedar Swamp 1,765 Neg. - -

Cx. pipiens (24) Stamford Cove Island Park 4,594 Pos. WNV(26) Cx. restuans (2)

Cx. pipiens (2) Stamford Sleepy Hollow Park 1,257 Pos. WNV(3) Cx. restuans (1)

Stamford Intervale Road 4,224 Pos. WNV(20) Cx. pipiens (20)

Stonington Barn Island 1,278 Neg. - -

Stonington Coogan Boulevard 557 Neg. - -

Stonington High Stonington 1,326 Neg. - - School

Stratford Beacon Point 3,265 Pos. WNV(5) Cx. pipiens (5)

Stratford Beaver Dam Road 894 Neg. - -

Tolland Bolton Lake 1,450 Neg. - -

Trumbull Cranbury Drive 635 Neg. - -

Pos. EEE(1) Cs. melanura (1) Voluntown Mt. Misery 3,541 Pos. JC(1) Oc. abserratus (1)

Wallingford South Elm Street 533 Pos. WNV(1) Cx. pipiens(1)

Waterbury Hamilton Park 699 Neg. - -

Pos. WNV(2) Cx. pipiens(2) Waterford Wtfd/N.London Line 2,039 Pos. JC(1) Oc. stimulans (1)

Cx. pipiens (7) West Hartford Spicebush Swamp 2,605 Pos. WNV(9) Cx. restuans (2)

Cx. pipiens (4) Pos. WNV (5) Cx. restuans (1)

West Haven Thill Street 6,530

Pos. JC(1) Oc. cantator (1)

Westbrook Willard Avenue 771 Neg. - -

Weston Devil’s Den 1,095 Neg. - -

Westport North Avenue 1,244 Neg. - -

Westport Sherwood Island 1,492 Pos. WNV(1) Cx. pipiens(1)

Wethersfield Goff Road 1,294 Pos. WNV(1) Cx. pipiens(1)

Willington Pinney Hill Road 609 Neg. - -

Wilton Saunders Drive 197 Neg. - -

Oc. aurifer (4) Wilton Spectacle Road 3,226 Pos. JC(6) Oc. canadensis(1) Oc. stimulans (1)

Windham Bass Road 3,561 Pos. JC(1) Oc. stimulans (1)

Woodbridge Burnt Swamp Road 1,103 Neg. - -

......

WNV(122) . Year to Date 173,988 - EEE (1) - JC(26)

APPENDIX M. Public Comment and Response

This appendix will be populated when the document is released for public comment.