1 Modern Threats to the Lepidoptera Fauna in The

Home , Ablerus, Acemya, Achatodes, Achatodes zeae, Acleris variana, Acoloithus

MODERN THREATS TO THE LEPIDOPTERA FAUNA IN THE FLORIDA ECOSYSTEM

THOMSON PARIS

A THESIS PRESENTED TO THE GRADUATE SCHOOL OF THE UNIVERSITY OF FLORIDA IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE

UNIVERSITY OF FLORIDA

2011

2011 Thomson Paris

To my mother and father who helped foster my love for butterflies

ACKNOWLEDGMENTS

First, I thank my family who have provided advice, support, and encouragement throughout this project. I especially thank my sister and brother for helping to feed and label larvae throughout the summer.

Second, I thank Hillary Burgess and Fairchild Tropical Gardens, Dr. Jonathan Crane and the University of Florida Tropical Research and Education center Homestead, FL, Elizabeth

Golden and Bill Baggs Cape Florida State Park, Leroy Rogers and South Florida Water

Management, Marshall and Keith at Mack’s Fish Camp, Susan Casey and Casey’s Corner

Nursery, and Michael and EWM Realtors Inc. for giving me access to collect larvae on their land and for their advice and assistance.

Third, I thank Ryan Fessendon and Lary Reeves for helping to locate sites to collect larvae and for assisting me to collect larvae. I thank Dr. Marc Minno, Dr. Roxanne Connely, Dr.

Charles Covell, Dr. Jaret Daniels for sharing their knowledge, advice, and ideas concerning this project.

Fourth, I thank my committee, which included Drs. Thomas Emmel and James Nation, who provided guidance and encouragement throughout my project.

Finally, I am grateful to the Chair of my committee and my major advisor, Dr. Andrei

Sourakov, for his invaluable counsel, and for serving as a model of excellence of what it means to be a scientist.

TABLE OF CONTENTS

page

ACKNOWLEDGMENTS ...... 4

LIST OF TABLES ...... 8

LIST OF FIGURES ...... 9

ABSTRACT ...... 13

CHAPTER

1 INTRODUCTION ...... 14

Threats to the Florida Ecosystem ...... 14 Project Objectives ...... 15

2 INVASIVE FIRE ANTS, SOLENOPSIS INVICTA AND NATIVE PLANTS IN FLORIDA ...... 16

Invasive Fire Ants ...... 16 Exotic Plants ...... 19

3 PARASITOIDS OF LEPIDOPTERA PARASITOIDS AND THEIR EFFECT ON NATIVE POPULATIONS IN FLORIDA...... 21

Biological Control Agents ...... 21 Biological Control ...... 21 Diptera: Tachinidae ...... 22 Objective ...... 23 Materials and Methods ...... 23 Natural History and Natural Enemies of Non-Target Species ...... 23 Natural History of Lepidoptera Species Surveyed for Parasitoidism in Florida in 2009-2010...... 24 Gulf Fritillary, Agraulis vanillae (Linnaeus) (Lepidoptera: Nymphalidae) ...... 24 Zebra Longwing, Heliconius charithonia (Linnaeus) (Lepidoptera: Nymphalidae)...... 26 Long-Tailed Skipper, Urbanus proteus (Linnaeus) (Lepidoptera: Hesperiidae) .....28 Dorantes Longtail. Urbanus dorantes (Stoll) (Lepidoptera: Hesperiidae)...... 30 Monarch, Danaus plexippus Linnaeus (Lepidoptera: Nymphalidae) ...... 31 Fall Webworm, Hyphantria cunea Drury (Lepidoptera: Arctiidae) ...... 36 Eastern tent caterpillar, Malacosoma americanum (Fabricius) (Lepidoptera: Lasiocampidae) ...... 40 Parasitoid Checklists for Six Additional Lepidoptera Hosts Collected During Survey ...... 43

Tawny emperor, Asterocampa clyton (Boisduval & Leconte) (Lepidoptera: Nymphalidae) ...... 43 Polydamas Swallowtail, Battus polydamas lucayus (Rothschild & Jordan) (Lepidoptera: Papilionidae) ...... 44 Mourning Cloak, Nymphalis antiopa (Linnaeus) Lepidoptera: Nymphalidae ...... 44 Brazilian Skipper, Calpodes ethlius (Stoll) (Lepidoptera: Hesperiidae) ...... 45 White-Marked Tussock Moth, Orgyia leucostigma (JE Smith) (Lepidoptera: Noctuidae) ...... 46 Survey Protocol ...... 47 Collecting Localities ...... 47 Collecting and Rearing ...... 47 Specimen Preservation and Identification ...... 48 Photography ...... 49 Results and Discussion ...... 49

4 DIFFERENCES BETWEEN HOST-PARASITOID RELATIONSHIPS IN URBAN AND RURAL SETTINGS ...... 66

Urbanization and Parasitoidism ...... 66 The Effect of Urbanization on Biodiversity ...... 66 Hypotheses for the Effect of Urbanization on Parasitoidism rates ...... 67 Objective ...... 69 Materials and Methods ...... 69 Habitat Types ...... 69 Tropical hammock ...... 69 Pine flatwood ...... 69 Freshwater marshes ...... 70 Disturbed habitat ...... 70 Beaches ...... 70 Mangrove ...... 71 Urban versus Rural Settings ...... 71 Data Analysis ...... 71 Results...... 73 Urban and Rural Settings ...... 73 Parasitoidism Ratios in Rural Versus Urban Habitats ...... 74 Gainesville, FL ...... 74 Miami, FL ...... 75 Discussion ...... 75

5 POSSIBLE EFFECTS OF PESTICIDES ON THE HOST-PARASITOID RELATIONSHIPS OF LEPIDOPTERA AND PARASITOIDS ...... 96

Pesticides and Parasitoids ...... 96 Are Pesticides Negative or Positive? ...... 96 Response of Caterpillars to Pesticides ...... 96 Effect of Pesticides on Parasitoids ...... 97 Materials and Methods ...... 97

Collection and Rearing ...... 97 Locations ...... 97 Results...... 99 Discussion ...... 100

6 DISCUSSION AND CONCLUSIONS ...... 113

APPENDIX: TACHINIDAE:DIPTERA OF FLORIDA AND THEIR HOSTS ...... 116

Description of Checklist ...... 116 Checklist of Diptera: Tachinidae in Florida and their Known Hosts ...... 117

LIST OF REFERENCES ...... 165

BIOGRAPHICAL SKETCH ...... 182

LIST OF TABLES

Table page

3-1 Life Table for Hyphantria cunea at NS1 in the First generation of 1967 (after Ito & Miyahsita 1968).Note: L I, LII, etc. mean the first-instar larva, second-instar larva, etc...... 50

3-2 Collection localities for survey of parasitoids in Florida...... 51

3-3 Taxa, whose larvae were collected and reared as part of parasitoid study in Florida in 2009-2010...... 54

3-4 Diversity of Parasitoids affecting Lepidoptera hosts...... 55

4-1 Habitat quantification for YMCA Road Gainesville, FL Scale: 3128 ft per 2 cm...... 80

4-2 Description of Habitat types...... 81

4-3 Sample sizes and parasitoid percentages (parasitoids/total adults eclosed) for each collection site in Gainesville and Miami, FL ...... 82

5-1 Parasitoid percentage in both pesticide and non pesticide settings in Alachua county. ..108

5-2 Parasitoid percentage and diversity in Pesticide and non-Pesticide settings in Florida counties Miami-Dade and Broward...... 111

5-3 Sample sizes and parasitoid percentages (parasitoids/total adults eclosed) for each collection site in Gainesville and Miami, FL ...... 112

LIST OF FIGURES

Figure page

3-1 Map of Gainesville with collection sites labled. See Table 3-1 for names of collection sites...... 52

3-2 Map of Miami, FL with collection sites labeled by letters. See Table 3-1 for names of collection sites...... 53

3-3 Chrysotachina sp. 1, voucher # 833, (Diptera: Tachinidae) a larval parasitoid of the Longtail Skipper Urbanus proteus (Lepidoptera: Hesperiidae)...... 56

3-4 Chrysotachina sp. 2,voucher # 147, (Diptera: Tachinidae)a larval parasitoid of the Longtail Skipper, Urbanus proteus (Lepidoptera: Hesperiidae)...... 56

3-5 Aplomya theclarum,voucher # 1467, (Diptera: Tachinidae)a larval parasitoid of the Appalachian Azure, Celastrina neglectamajor (Lepidoptera: Lycaenidae)...... 57

3-6 Tachinidae sp. 4.voucher # 805, (Diptera: Tachinidae)a larval parasitoid of theGulf Fritillary, Agraulis vanillae (Lepidoptera: Nymphalidae)...... 57

3-7 Tachinidae sp. 5, voucher # 807, (Diptera: Tachinidae) a larval parasitoid of theDorantes Longtail, Urbanus dorantes (Lepidoptera: Hesperiidae)...... 58

3-8 Tachinidae sp. 6, voucher # 1460, (Diptera: Tachinidae) a larval parasitoid of theMonarch, Danaus plexippus (Lepidoptera: Nymphalidae) ...... 58

3-9 Tachinidae sp. 7, voucher # 1168, (Diptera: Tachinidae)a larval parasitoid of the: Eastern Tent Caterpillar Moth, Malacosoma americanum (Lepidoptera: Lasiocampidae)...... 59

3-10 Tachinidae sp. 8, voucher # 1148, (Diptera: Tachinidae) a larval parasitoid of host White-marked Tussock Moth, Orgyia leucostigma (Lepidoptera: Lymantriidae)...... 59

3-11 Tachinidae sp. 9,voucher # 1337, (Diptera: Tachinidae)a larval parasitoid of the Mourning Cloak, Nymphalis antiopa (Lepidoptera: Nymphalidae)...... 60

3-12 Tachinidae sp. 10.voucher # 807, (Diptera: Tachinidae) a larval parasitoid of theDorantes Longtail,Urbanus dorantes (Lepidoptera: Hesperiidae), ...... 60

3-13 Campopleginae sp. 1,voucher # 3019, (Hymenoptera: Ichneumonidae) a larval parasitoid of the Eastern Tent Caterpillar, Moth, Malacosoma americanum (Lepidoptera: Lasiocampidae) ...... 61

3-14 Campopleginae sp. 2,voucher # 1470, (Hymenoptera: Ichneumonidae) a larval parasitoid of theAppalachian Azure, Celastrina neglectamajor (Lepidoptera: Lycaenidae)...... 61

3-15 Chalcidoidea sp. 1, voucher # 1321, (Hymenoptera: Chalcidoidea) a larval parasitoid of theTussock moth (Lepidoptera: Lymantriidae) ...... 62

3-16 Mesochorinae sp 1, voucher # 981, (Hymenoptera: Ichneumonidae) a larval parasitoid of the Fall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae)...... 62

3-17 Braconidae sp. 1, voucher # 1046, (Hymenoptera: Braconidae) a larval parasitoid of the Fall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae)...... 63

3-18 Chalcidoidea sp. 1, voucher # 1101, (Hymenoptera: Chalcidoidea) a larval parasitoid of the Fall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae)...... 63

3-19 Chalcidoidea sp. 2, voucher # 1453, (Hymenoptera: Chalcidoidea) an egg parasitoid of the Cecropia silkmoth, Hyalophora cecropia (Lepidoptera: Saturniidae)...... 64

3-20 Chalcidoidea sp. 3, voucher # 1454, (Hymenoptera: Chalcidoidea) a larval parasitoid of theFall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae)...... 64

3-21 Chalcidoidea sp. 4, voucher # 1510, (Hymenoptera: Chalcidoidea) a larval parasitoid of theGulf Fritillary,Agraulis vanillae (Lepidoptera: Nymphalidae)...... 65

3-22 Chalcidoidea sp. 5, voucher # 2402, (Hymenoptera: Chalcidoidea) a parasitoid of the Gulf Fritillary, Agraulis vanillae (Lepidoptera: Nymphalidae)...... 65

4-1 Land usage in the United States from 1960 to 2000 (after Stein et al. 2000)...... 78

4-2 Habitat quantification via Satellite Images. August 1, 2010, Google Earth, Scale: 3130 ft. per 2 cm, 12 cm x 18 cm (total area 216 cm2) Total Area: 18.98 mi2 ...... 79

4-3 Comparison and contrast of the range of percentages of three characters (streets, buildings, and trees/shrubs) found in thirteen collection areas...... 83

4-4 Habitat quantification for YMCA Road Dec 17, 2007, Google Earth, Scale 3128 ft (. 216 cm2, 12 cm x 18 cm) ...... 84

4-5 Habitat quantification for SW 63rd AveDec 17, 2007, Google Earth, Scale 3130 ft. (216 cm2, 12 cm x 18 cm) ...... 84

4-6 Habitat quantification for 301 Shell station Dec 17, 2007, Google Earth, Scale 3134 ft. (216 cm2, 12 cm x 18 cm) ...... 85

4-7 Habitat quantification for Stadium by pool Dec 17, 2007, Google Earth, Scale 3131 ft. (216 cm2, 12 cm x 18 cm) ...... 85

4-8 Habitat quantification for NATL Dec 17, 2007, Google Earth, Scale 3134 ft. (216 cm2, 12 cm x 18 cm) ...... 86

4-9 Habitat quantification for NW 98 St Dec 17, 2007, Google Earth, Scale 3128 ft. (216 cm2, 12 cm x 18 cm) ...... 86

4-10 Habitat quantification for Santa Fe Dec 17, 2007, Google Earth, Scale 3131 ft. (216 cm2, 12 cm x 18 cm) ...... 87

4-11 Habitat quantification for Kanapaho Park Dec 17, 2007, Google Earth, Scale 3128 ft. (216 cm2, l12 cm x 18 cm) ...... 87

4-12 Habitat quantification for Everglades Holiday Park April 1, 2010, Google Earth, Scale 3135 ft. (216 cm2, 12 cm x 18 cm) ...... 88

4-13 Habitat quantification for South Florida Water Management April 1, 2010, Google Earth, Scale 3130 ft. (216 cm2, 12 cm x 18 cm) ...... 88

4-14 Habitat quantification for Casey's Corner Nursery August 1, 2010, Google Earth, Scale 3133 ft. (216 cm2, 12 cm x 18 cm) ...... 89

4-15 Habitat quantification for UF Tropical Research Station August 1, 2010, Google Earth, Scale 3133 ft. (216 cm2, 12 cm x 18 cm) ...... 89

4-16 Habitat quantification for Abandoned Plot August 1, 2010, Google Earth, Scale 3130 ft. (216 cm2, 12 cm x 18 cm) ...... 90

4-17 Habitat quantification for Bill Baggs Cape Florida State Park April 1, 2010, Google Earth, Scale 3128 ft ...... 90

4-18 Percentage of parasitoids (Diptera: Tachinidae) affecting hosts Agraulis vanillae in urban and rural settings in Gainesville, FL from October to December 2009 ...... 91

4-19 Percentage of parasitoids (Diptera:Tachinidae and *Hymenoptera:Ichneumonidae) affecting hosts Hyphantria cunea in urban and rural settings in Gainesville, FL in July 2010 ...... 92

4-20 Comparison of parasitoids affecting Hyphantrea cunea in April and July in rural Gainesville, FL...... 93

4-21 Percentage of parasitoids (Diptera: Tachinidae) affecting hosts Urbanus proteus in urban and rural settings in Gainesville, FL from September to October in 2009 and from August to September in 2010 ...... 94

4-22 Percentage of parasitoids (Hymenoptera: Chalcidae) affecting hosts Agraulis vanillae in urban and rural settings in Miami, FL from April to September in 2010...... 95

5-1 Map of Gainesville with pesticide spray zones...... 102

5-2 Map of Miami with 2009 and 2010 pesticide spray zones...... 103

5-3 Percentage of parasitoids (Diptera: Tachinidae) affecting hosts Agraulis vanillae in pesticide and non-pesticide settings in Gainesville, FL from October to December 2009...... 104

5-4 Percentage of parasitoids (Diptera: Tachinidae) affecting hosts Urbanus proteus in pesticide and non-pesticide settings in Gainesville, FL from September to October in 2009 and from August to September in 2010 ...... 105

5-5 Percentage of parasitoids (Diptera:Tachinidae and*Hymenoptera:Ichneumonidae) affecting hosts Hyphantria cunea in pesticide and non-pesticide settings in Gainesville, FL in July 2010 ...... 106

5-6 Percentage of parasitoids (Hymenoptera: Chalcidae) affecting hosts Agraulis vanillae in pesticide and non-pesticide settings in Miami, FL from April to September in 2010...... 107

Abstract of Thesis Presented to the Graduate School of the University of Florida in Partial Fulfillment of the Requirements for the Degree of Master of Science

MODERN THREATS TO THE LEPIDOPTERA FAUNA IN THE FLORIDA ECOSYSTEM

Thomson Mason Paris

May 2011

Chair: Andrei Sourakov Major: Entomology and Nematology

In the present study, I examine the possible threats to the Lepidoptera fauna of Florida.

Factors affecting the Lepidoptera fauna in Florida ecosystems include native and exotic parasitoids. Over 3,000 larvae of Lepidoptera were sampled in Alachua, Broward, and Miami-

Dade Counties in August 2009 -September 2010. The larvae were reared in the lab. To-date, a

total of 596 parasitoids were obtained from the rearings. The survey provided information about

the prevalence of parasitoids on select native species and populations of Lepidoptera in Florida.

The differences between host-parasitoid interactions in urban and rural settings and the possible

effect of pesticides on these interactions are reported.

CHAPTER 1 INTRODUCTION

Threats to the Florida Ecosystem

Florida has significant pressure on its ecosystem due to a variety of factors that include urbanization, pesticide and herbicide usage, and the introduction of nonnative species. These factors have led to some anecdotal reports of a decrease in Florida Lepidoptera species, especially in South Florida. Several isolated refuges within the city limits of certain south Florida cities (for example, Castellow Hammock Park and Camp Owaissa Park) seem to possess a greater butterfly diversity than large tracts of protected land (for example, Biscayne National

Park and Everglades National Park) (Minno & Minno 2009). There are anecdotal observations that butterfly diversity might be higher in relatively urban areas that are sprayed for mosquitoes than in the unsprayed areas (Daniels pers. comm. 2010; Minno & Minno 2009).

The impact of urbanization on an ecosystem is not only species extermination, but also the alteration of the balance of the ecosystem. Florida experiences a high influx of exotic species

(Frank & McCoy 1995). Biological control agents are one type of nonnative species introduced into the Florida ecosystem (Frank & McCoy 1995). While their negative effect on the native species has not been found in Florida, further north, the introduced parasitoid Compsilura concinnata (Diptera: Tachinidae) has had a negative impact on the local Saturniidae population

(Boettner et al.2000, Kellog et al.2003). In 1993, Frank and McCoy published a list of all known introduced insects into Florida, which included 351 invertebrates (1993). One hundred and fifty four were reported as being established in Florida (Frank & McCoy 1993). By excluding the list of established insects to those that were only introduced for biological control, Frank and McCoy reduced the list to 60 invertebrates (2007). After a thorough examination of biological records of each of the 60 species, 10 were found to be a potential risk to non-target populations. None of the

ten detrimental biological agents were found to have had a marked negative effect on non-target

populations (Frank & McCoy 2007).

Project Objectives

The influence of factors like parasitoids and urbanization on the Florida ecosystem is the focus of this study. Both direct and indirect effects are evaluated in this study. Indirect effects with respect to urbanization involve factors,such as presence of exotic parasitoids and fire ants, that have established in new niches due to urbanization. These non-natives outcompeted old occupants in recently formed urban habitats. Indirect effects also include the impact of pesticides on insects, accidental invaders (fire ants), exotic plants, and native parasitoids as well as nonnative parasitoids. Some parasitoids might have been introduced intentionally, to control the accidental invaders, such as Lymantria dispar (Linnaeus) (Lepidoptera:Lymantriidae). Direct effects include pesticides usage and intentional habitat destruction during development. The goal of my project is to explore possible variation in parasitism of native butterfly species caused by native and exotic parasitoids in a variety of habitats throughout Florida, with special emphasis on urban and rural areas, as well as areas with heavy versus little or no mosquito spraying.

Additionally, I am providing a comprehensive review of scientific literature pertaining to harmful effects of introduced arthropods on Florida Lepidoptera populations.

CHAPTER 2 INVASIVE FIRE ANTS, SOLENOPSIS INVICTA AND NATIVE PLANTS IN FLORIDA

Invasive Fire Ants

The state of Florida contains the highest number of exotic ants (52 species) in the United

States (Deyrup et al. 2000). The exotic ants have varying degrees of impact on Florida ecosystems. Several of the more intrusive and devastating species to Florida ecosystems include

Solenopsis invicta Buren (Hymenoptera: Formicidae), Pseudomyrmex gracilis (Fabricius)

(Hymenoptera: Formicidae), Camponotus planatus (Roger) (Hymenoptera: Formicidae),

Wasmannia auropunctata (Roger) (Hymenoptera: Formicidae) (Deyrup et al. 1984 2000). Exotic

ants were introduced to Florida initially via the importation of exotic plants and as stowaways in

cargo ships. Because of higher monitoring efforts of plants imported to the US, more recent

exotic ant invasions have occurred via airplanes (Deyrup et al. 2000).

One of the most successful invaders to enter the United States, accidentally, is the Red

Imported Fire Ant, Solenopsis invicta. Approximately 50 years after its introduction into the

United States, S. invicta inhabits 150 million hectares (Meer et al. 1986). The origin of S. invicta

in the United States was Mobile, Alabama around 1933 via a cargo ship. Solenopsis invicta’s

establishment of a beachhead in Mobile, Alabama was preceded by the arrival of its close

relative, the Black Fire Ant, Solenopsis richteri Buren (Hymenoptera: Formicidae), in 1928

(Tschinkel 2006). Solenopsis invicta was spread by the shipment of plant material to a variety of

nurseries and by 1953 it had been identified in 10 states and 109 counties in the southeastern part

of the United States (Tschinkel 2006). King and Tschinikel (2008) aptly point out that S. invicta

is not a driver of ecological change but merely a passenger. Solenopsis invicta’s success upon

arriving in new territories stems from its high reproductive capacity. Each colony of fire ants can produce 3 to 5 thousand queens per colony per year (Lofgren and Meer 1986). Further

enhancing S. invicta’s ability for colonization is the fact that many of its natural enemies, such as parasitoids and pathogens, are lost upon colonization (Yang et al. 2010).

As S. invicta has multiplied and expanded across the southeastern region of the United

States, questions have arisen regarding its impact on newly invaded ecosystems. In some cases,

S. invicta has been shown to actually reduce the population of some pest species, such as, the boll weevil, Anthonomus grandis grandis Boheman (Coleoptera: Curculionidae), the horn fly,

Haematobia irritans (Linnaeus) (Diptera: Muscidae) , and the tobacco budworm, Heliothis virescens (Fabricius) (Lepidoptera:Noctuidae) (Wojcik 2001). Solenopsis invicta is relatively nonspecific in it prey selection, which has had a detrimental effect on the beneficial arthropod communities where S. invicta has invaded (Eubanks et al. 2002; Forys et al. 2001; Morrison

2002; Porter et al. . 1990; Risch 1982; Vinson 1990; Wojcik 2001). Allen et al. (2001) found that a general reduction in the arthropod population and subsequent reduction in the population of the

Loggerhead shrike, Lanius ludovicianus Linnaeus (Passeriformes: Laniidae), was linked to the presence of S. invicta. Other studies point to S. invicta having a more moderate effect on the ecosystem (Morrison & Porter 2003). It appears, that the initial negative effects of S. invicta in a newly invaded ecosystem may become more moderate over time (Morrison 2002). Morrison found that otherwise identical plots that possess S. invicta had greater ant and arthropod diversity then those plots that possessed less S. invicta (2002).

The negative impacts of S. invicta on Lepidoptera in Florida have been reported in several studies (e. g., Kenis et al. 2009). Forys et al.estimated the potential negative effect of S. invicta on Heraclides aristodemus ponceanus (Schaus) (Lepidoptera: Papilionidae) by studying the egg, larvae, and pupa life stages of Heraclides cresphontes (Cramer) (Lepidoptera: Papilionidae) in the Florida Keys. The investigators chose H. cresphontes as their study model forH. a.

ponceanus (Schaus) because both feed on plants in the family Rutaceae and both are members of

the same lepidotperan family, Papilionidae. The investigators concluded that S. invicta was likely

to be endangering the survival of H. a. ponceanus in the Florida Keys (Forys et al. 2001). Further research is needed directly with H. a. ponceanus and S. invicta to be able to make more conclusive statements regarding S. invicta’s affect on H. a. ponceanus.

The interaction between the foraging S. invicta at each of the three life stages (eggs, larvae, and pupae) of H. a. ponceanus could be variable and perhaps even at some stage beneficial. For example, the Cyclargus thomasi bethunebakeri Comstock & Huntington was also found to be at risk from predation by S. invicta, but after further study was observed receiving ant tending behaviors in the field (Trager & Daniels 2009).

Solenopsis invicta has hampered Lepidoptera larvae from becoming successfully established as biological control agents against invasive water lettuce, Azolla caroliniana Willd.

(Azollaceae: Salviniales) (Cuda et al. 2007; Cuda et al. 2004; Dray Jr. 2001).Stemming the negative effect of Solenopsis invicta has been attempted through the use of pesticides. Bacillus thuringiensis Berliner pesticides were used to control S. invicta in an area where Lepidoptera were being used as biological control agents to control an invasive aquatic weed Azolla carolinana Willd. (Azollaceae: Salviniales). Following the application of the pesticide, and subsequent reduction in the population of S. invicta, the Lepidoptera biological control agents were able to flourish and successfully control the invasive weed (Cuda et al. 2004).

Meer et al. (1986) promote the benefits of an integrated pest management approach for the control of S. invicta 1986). As a method for management, Forys et al. (2001) found that insecticide treatments using Andro for S. invicta were ineffective. Instead of insecticides, cultural practices such as habitat restoration of the Florida Keys hardwood hammock were cited as the

preferred long-term method for S.invicta’s control in the Florida Keys (Forys 2001). Forys et al.

(2001) found that there was a greater abundance of S. invicta where the habitat had undergone

modification, which included paths, roads, and powerlines.

Exotic Plants

As in the case with non-native insects, the introduction of non-native plants has resulted in changes to the ecosystems in south Florida. There are several examples of Lepidoptera benefitting from the exploitation of introduced plants. Erynnis baptisiae (Forbes) (Lepidoptera:

Hesperiidae) has made a transition from its traditional host Wild Indigo, Baptisia tinctoria

(Linnaeus) (Fabales: Fabaceae), to an invasive plant Crown Vetch, Securigera varia (Linnaeus)

(Fabales: Fabaceae),a plant originally used to stem soil erosion along roadsides (Department of

Conservation and Recreation). Staphylus hayhurstii (Edwards) (Hesperiidae: Lepidoptera), has expanded its range to the north, because of the introduction of Lamb’s quarters, Chenopodium album Linnaeus (Caryophyllales: Amaranthaceae). Chenopodium album was introduced into the

United States accidentally through contaminated agricultural seed packets (Schweintz 1832;

Mack & Erneberg 2002). Despite the above examples, most of the exotic plant introductions to the United States have not benefited butterflies. Some plant introductions have been detrimental to butterflies. Garlic weed, Alliaria petiolata (Bieb.) Cavara & Grande (Brassicales:

Brassicaceae), has chemical markers that induce a higher attractiveness to the West Virginia white butterfly, Pieris virginiensis Edwards (Lepidoptera: Pieridae:) than its typical host plants

Dentaria diphylla and Dentaria laciniata (Brassicales: Brassicaceae) (Cech and Tudor 2005).

Alliaria petiolata was probably introduced into the United States by settlers as a food or medicinal crop (Nuzzo 1993). Pieris virginiensis larvae that develop on this exotic host die by the 1st or 2nd instar. Likewise, Vincetoxicum nigrum Kartesz & Gandhi(Gentianales:

Asclepiadaceae) causes mortality to larvae of Danaus plexippus(Linnaeus) (Nymphalidae:

Lepidoptera) (Casagrande & Dacey 2001; Haribal & Renwick 1998). Vincetoxicum nigrum,

which outcompetes native milkweeds, was first introduced into the United States from Europe in

gardens as a weed, in 1882 between New England and Pennsylvania (Sheely & Dudley 1996).

The train of invasive species starts with Chinese tallow, Sapium sebiferum, Melaleuca,

Melaleuca quinquenervia, Brazilian pepper-tree, Schinus terebinthifolius, Australian pine,

Casuarina equisetifolia, Chinaberry, Melia azedarach,and mimosa tree, Albizia julibrissin etc.

CHAPTER 3 PARASITOIDS OF LEPIDOPTERA PARASITOIDS AND THEIR EFFECT ON NATIVE POPULATIONS IN FLORIDA

Biological Control Agents

Biological Control

Unlike the accidental introduction of ants, many biological control agents have been purposefully introduced into Florida over the past hundred years. Frank and McCoy (1993) report that of the 351 potential biological control agents brought into Florida, 154 were released into the wild. Of the insects released, 24.5 % were targeting insect pests, the remainder targeted weedy plants. Of the total number of insect pests targeted, 24% were Lepidoptera (Frank &

McCoy 1993). In an analysis of the potential risk of classical biological control, it was found that

24 agents could threaten non-target species of insects in their host range. Of these 24 agents, 10 agents were implicated to have caused population changes in non-target species and four agents were shown to affect non-target species populations (Frank & McCoy 2007). These four species included Cotesia flavipes Cameron (Hymenoptera: Braconidae), Aphytis holocanthus DeBache

(Hymenoptera: Aphelinidae), Coccinella septempunctata DeBach (Coleoptera: Coccinellidae),

Cryptolaemus montrouzieri Mulsant (Coleoptera: Coccinellidae) (Frank & McCoy 2007).

The ability of exotic species to exploit native arthropod communities is evident in Hawaii

(Kaufman & Wright 2009). Of the parasitoids affecting Lepidoptera larvae collected in the field in Hawaii, 84% were biological control agents. Among the larvae that were collected in the field in Hawaii, as much as 21% were parasitized (Henneman & Memmott 2001). It should be noted that all of these biological control agents were generalists released before 1945. It is important to note that following 1945, in Hawaii, there was a significant change in biological control practices from releasing generalist parasitoids to only releasing specialist parasitoids into the targeted ecosystem (Henneman & Memmott 2001).

The biological control agent, Compsilura concinnata (Meigen) (Diptera: Tachnidae) was released to control 13 pest species among which was the gypsy moth, Lymantria dispar

(Linnaeus) (Lepidoptera: Lymantriidae) in New England in 1918 (Boettner et al.2000). As in the

biological control agents of Hawaii, Compsilura concinnata is a generalist parasitoid and

developed a non-target host range, which included silk moths, family Saturniidae. The effects of

this newly formed parasitoid-host relationship have been linked to the marked decline of several

species of silk moths in New England (Boettner et al. 2000). Kellog et al. (2003) found

parasitism by the parasitoid C. concinnata of Actias luna Linnaeus (Lepidoptera: Saturniidae)

larvae to be between zero and 62%.

Diptera: Tachinidae

Though parasitic Diptera in general are not considered to be as effective parasites as

parasitic Hymenoptera (Askew 1971; Quicke 1997), there are more than 10,000 species of

Tachinidae in the world (O’Hara 2008), which are capable of parasitizing a wide array of insect

hosts. As a result, Tachinidae have been used as biological control agents to help stem the

growth of unwanted pests without the use of pesticides. Such practices have on occasion resulted

in unanticipated negative effects towards non-target species. Compsilura concinatta is a biological control agent introduced into New England to help control the Gypsy moth, but has been found to have adverse effect on the populations of several species of moths in the family

Saturniidae (Boettner 2000; Kellog 2003).

Little is known about the Neotropical and Australasian Tachinidae. Illustrating this fact was a study carried out in Costa Rica, which divided a set of 17 seemingly generalist parasitoids into 32 highly host specific cryptic species through the aid of DNA barcoding (Smith et al.

2006). O’Hara estimates that 5,000 species of Tachinidae are undescribed in the neotropics and

Australasian regions (O’Hara 2008).

While the Neotropical and Australian regions of the world have great gaps in the knowledge of Tachinidae, the Nearctic region has benefited from the work of specialists such as

James O’Hara and Paul Arnaud. Arnaud (1978) compiled an exhaustive set of data comprising all the known host species for Tachinidae in North America up to 1973. O’Hara and Wood report that 261 species of Tachinidae are found in Florida alone (2004) (see Appendix for more information).

Objective

The present study attempts to discern the effect of native and introduced parasitoid species

on native species of Lepidoptera in Florida in a variety of natural and man-made ecosystems.

This study consisted of collecting Lepidoptera larvae in several different localities in Florida,

rearing them through on their hostplants while collecting data on ratio of parasitism, as well as

voucher specimens of resulting parasitoids

Materials and Methods

Natural History and Natural Enemies of Non-Target Species

As a preparations for the parasitoid survey, parasitoid records for the thirteen species of

Lepidoptera listed in Table 3-3 were found by consulting Krombein et al.(1979) for

Hymenopteran parasitoids and Arnaud (1978) for Tachinid parasitoids. A literature search in

Google Scholar was conducted to obtain more recent parasitoid-host records. Seven Lepidoptera

species were surveyed and hence a short description that includes the biology, life cycle, host

plants, and parasitoid predators is provided below for each of these species. The Lepidoptera

species whose larvae were only occasionally collected, are listed further, with only a checklist of

their parasitoids provided.

Natural History of Lepidoptera Species Surveyed for Parasitoidism in Florida in 2009- 2010.

Gulf Fritillary, Agraulis vanillae (Linnaeus) (Lepidoptera: Nymphalidae)

The common name, Gulf Fritillary, is derived from frequent observations of this species flying over the Gulf of Mexico (Pyle 1981). Agraulis vanillae is a species that is capable of surviving in and near urban centers (Wagner 2005). This fact is related to A. vanillae’s preference for disturbed habitats that contain its host, Passiflora, and nectar sources. During

migration, it can be found in almost any habitat (Cech & Tudor 2005). A. vanillae is distributed

throughout Florida and the southeast (Kimball 1965). Agraulis vanillae nigrior is the typical

variety encountered in the east (Cech & Tudor 2005). In 1932, a subspecies of A. vanillae was

found, in Key Largo, called Agraulis vanillae comstocki (Forsyth 1932). Although isolated

forms, such as Agraulis vanillae comstocki, exist, A. vanillae’s habit of migration precludes it

from developing discernable forms (Cech & Tudor 2005).

Distribution: A. vanillae is seen year round in parts of Florida and Texas (Wagner 2005).

Adult strays have extended its northward range into Ohio, New York, and Pennsylvania (Cech &

Tudor 2005; Whan & Belth 1992).

Description: The egg is oval in shape. The surface is not smoothe, but has columnar ridges

rising from the base to the crest of the egg. At the crest, the egg is flattened and the small

protuberances form a circle. The egg is creamy yellow.

The larva is bright orange with ribbons of brown. It can reach 1.5’’ in length (Minno et

al. 2005). Wagner reports that Texas varieties of A. vanillae have ribbons of purple instead of

brown (2005). Large black spines also extend out from the larval body.

The pupa is light brown with splotches of white and darker brown. A few spots of silver are on the back. The chrysalis appears to resemble a bird dropping and/or a withered leaf

(Wagner 2005).

The adult is bright orange on the dorsal part of the forewing and hindwing. The tips of the

wings have black spots. The ventral view of the hind wings and forewings reveals large silver

spots. Males have a pheromone circulating structure on multiple veins of the dorsal forewing

(Rauser & Rutoski 2003).

Biology: The life cycle of A. vanillae begins with the female ovipositing a single egg on a

Passiflora spp. The author has observed the female laying an egg adjacent to, but not on the host.

Hosts Plants:

• Passiflora incarnata; • Passiflora suberosa; • Passiflora multiflora; and • Other Passiflora species.

The aposematically colored larvae are toxic to predators (Cech & Tudor 2005), while the adults are preyed upon by Grossbeaks and Orioles (Brower 1985) and probably other birds.

More recent studies indicate that abdominal glands present in both sexes of adult A. vanillae may be defense against predation (Ross et al. 2001).

The pupa stage follows the larval stage. Pupation has been noted as far as fifty feet from the nearest host plant (Kimball 1965). It has been thought that in the southern United States, A. vanillae overwinter as adults (Cech & Tudor 2005). Surviving winter, however, may be accomplished during the pupal and larval stages in north-central Florida (Sourakov 2009). While

the immatures of A. vanillae may be able to endure freezing temperatures, adults undergo

seasonal migrations. In Florida, adults can be observed flying north in the spring (between

February and June) and south in the fall (between August and November) (Walker 1978; 1991;

2001). Numbers of migrating Lepidoptera, including Agraulis vanillae, flying north in the spring

have been estimated to be four million and, in the fall, 40 million (Walker 1991).

Agraulis vanillae males seek to copulate with females that have recently emerged, are not

yet able to fly, or are still partially in the pupa (Tveten, J. & Tveten, G. 1996).

Natural Enemies: Several types of birds have been observed eating adult A. vanillae, although this seems to rarely occur in the wild, which include Grossbeaks and Orioles (Brower

1985; Ross et al.2001).

Parasitoids known to use A. vanillae as a host are:

Hymenoptera Chalcidoidea Chalcididae Chalcis flavipes (Fabricius) Pteromalus puparum Linnaeus (Krombein 1979)

Diptera Oestroidea Tachinidae Compsilura concinnata (Meigen) Arnaud (1969) Chetogena claripennis (Macquart) Arnaud (1978) Lespesia aletiae (Riley) Hyphantrophaga virilis (Aldrich & Webber), undetermined Hyphantrophaga sp. (Sourakov 2009)

Zebra Longwing, Heliconius charithonia (Linnaeus) (Lepidoptera: Nymphalidae).

Heliconius charithonia is the state butterfly of Florida (Cech & Tudor 2005). It is fitting

that Florida chose H. charithonia, because it was the first butterfly described in Florida (Bartram

1996). It is one of the longest living butterflies, which may be due to its habit of pollen feeding

(Gilbert 1972).

Distribution: Heliconius charithonia is a widespread species extending from South

America to southern parts of Georgia. Adults may wander as far north as New York (Cech &

Tudor 2005). It can also be seen year round in South Florida and Texas (Wagner 2005).

Description: The egg is oval in shape and has ridges that proceed upward in columns. The

egg is blunt at the crest and the ridges form circles. It is yellow in color.

The larvae are white with black spines that may reach up to 1.7” in length. The white body

is interspersed with black dots. The head of the larva contains two black horn-like spines that

project forward.

The pupa is light brown with dark brown splotches and silver spots. The pupa is shaped

like a sea horse. The small jaws and reflecting silver spots give the pupa a menacing appearance

to potential prey (Minno, M. C. & Minno, M. 1999).

Adults have forewings that extend out beyond the width of the hind wing. The wings are

chiefly black in color and have long thin stripes of yellow that extend across both the hind wing

and the forewing. The ventral parts of adults have small red dots close to the thorax that is on

both the hind wings and forewings.

Biology: Heliconius charithonia females oviposit eggs laid in clusters on new growth in

shady areas on a variety of Passiflora spp.

Host plants:

• Passiflora incarnata; • Passiflora lutea; • Passiflora suberosa; and • Passiflora multiflora (Minno, M.C. & Minno M. 1999; Cech & Tudor 2005).

The longevity of the adults is linked to their ability to include pollen in their diet. Pollen consumption is achieved by the secretion of enzymes from the proboscis, allowing adequate digestion (Cech & Tudor 2005; Wagner 2005).

Like many other Heliconius spp., H. charithonia exhibits pupal mating. The only description of which is based on observations conducted in Gainesville, FL. Males began congregating on the pupa days prior to the female becoming pharate. During this anticipatory

period fights between rival males took place. Male copulation occurred two to three hours prior

to the eclosion of the female (Sourakov 2008).

Another unique behavior among the Heliconius spp. is group sleeping. Adults sleep in groups of 25 to 30 called crèches (Cech & Tudor 2005). The benefit of the crèches formation

include (a) shelter, (b) a drier area compared to the surrounding habitat, and (c) reduced light

conditions (Salcedo 2010).

Natural Enemies: After 22 surveys, egg parasitism for H. charithonia was found to

average 53.0 ± 5.0% with an average of 6.6 ± 0.6 wasps (n=34; range: 1-14) (Fleming et

al.2005).

Diptera Oestroidea Tachinidae Unidentified Tachinidae larva (Quintero 1988) Hymenoptera Chalcidoidea Trichogrammatidae unidentified Trichogrammatid wasp. (Fleming et al. 2005)

Long-Tailed Skipper, Urbanus proteus (Linnaeus) (Lepidoptera: Hesperiidae)

Urbanus proteus or “leaf roller”, as it is commonly called, is a migratory species that

occurs in the southern region of the United States. Its range is limited by its inability to tolerate

freezing temperatures (Opler 1992). From the mid 1800’s to the mid 1900’s adults were

observed in parts of the northeast United States, after which, adults were not observed for about

forty years in the northeast. Perhaps, as a result of additional soy bean fields, adults were again

observed in the northeast during the 1990s (Cech & Tudor 2005).

Distribution: Urbanus proteus is a year round resident in parts of Florida and Texas.

During the summer months, adults venture as far north as New England, Kansas, and Illinois.

The planting of soy beans in various regions seems to be related to its distribution (Walker 1978;

Cech & Tudor 2005).

Description:The egg is laid singly or stacked on two or three other eggs (pers. observation; Greeney & Sheldon 2008). The egg is circular with smooth ridges forming 10 or 11 sectors and yellow in color.

The larva grows to 1.5” in length (Minno et al. 2005) and is made up of several shades of green, yellow, and black. Two yellow stripes sandwiched between a black stripe run along the dorsal surface of the larva. The head capsule is reddish brown, except for the area on the front that has a black spot. Two “eyespots” by the stemmata are orange.

The pupa is brown and covered with a white dust that acts as a waterproofing agent (Scott

1992).

The adult is brown and possesses metallic blue green on the forewings and hind wings near the body. Semitransparent spots are present on the forewing and the hind wing has a long tail. Males possess a row of androconial scales along the costal margin of the forewing.

Biology: As noted above, females oviposit 1 to several eggs (>1 = stacked) on a wide variety of host plants in the pea family (Minno et al.2005). Larvae form three to five different leaf shelters during their five instar larval cycle (Greeney & Sheldon 2008).

Adults have been observed migrating north in the spring and south in fall (Walker 1978;

2001). In the spring and summer, less adults are observed in Florida (Kimball 1965). In fall,

there is an increased amount of adults observed, which could be due to an increase in the

abundance of soybean farms in the southeast (Cech & Tudor 2005).

Natural Enemies: Predators of U. proteus include many spp. of Polistes (Hymenoptera:

Vespidae) and Euthyrhynchus floridanus (Linnaeus) (Hemiptera: Pentatomidae). Another

significant factor effecting mortality a nuclear polyhedrosis virus has been reported. Its

infestation has been noted to result in the death of as many as 40 to 50 % of fall larvae in one

study (Capinera 2007).

Diptera Oestroidea Tachinidae Lespesia aletiae (Riley) Nemorilla pyste (Walker) (Arnaud 1978) Chrysotachina alcedo (Loew) (Arnaud 1978; Capinera 2007)

Dorantes Longtail. Urbanus dorantes (Stoll) (Lepidoptera: Hesperiidae)

Urbanus dorantes was recently introduced to Florida in 1969 (Knudson 1974). The origin of this introduction was shown to be Texas and not the Caribbean due to the immense sea barrier required to originate directly from the Antilles would be unlikely (Miller, L. & Miller, J. 1970).

The multiple shared characteristics between Urbanus proteus and Urbanus dorantes give rise to questions regarding the mechanisms involved in the process of speciation (Cech & Tudor 2005).

Distribution: In the United States Urbanus dorantes is regularly found in Florida, southern Texas, and southern Arizona (Minnoet al. 2005). Occasionally, Urbanus dorantes strays north to North Carolina and Virginia, while its southern border is Argentina (Cech &

Tudor 2005).

Description: The egg is circular in shape and has ridges that divide the egg up into 10 or

11 sectors. The crest of the egg is blunt and the ridges form a circular ridge. It is light green in color.

The larva is either orange and yellow with brown spots or green and yellow with brown

spots. There is a green stripe that runs from the head to the end of the abdomen on the dorsal

surface of the body. The head capsule is uniformly brown, which distinguishes U. dorantes

larvae from Urbanus proteus larvae (Wagner 2005).

The pupa is brown with scattered dark brown spots.

The adult is very similar to Urbanus proteus, but it lacks the blue green metallic scales near the body. Semitransparent spots are present on the forewing and the hind wing has a long

tail.

Biology: A single egg is oviposited by the female on the host (Minno, M. C. & Minno, M.

1999). Adults produce several broods per year. It differs from Urbanus protesus, in that, it is not

a migrating species. Therefore, parts of Florida have Urbanus dorantes adults year round

(Minno, M. C. & Minno, M. 2005).

Natural Enemies: Hymenoptera Ichneumonoidea Ichneumonidae *Ichneumon panamensis Cameron *Trogomorpha arrogans (Cresson) *Trogomorpha trogiformis (Cresson) *Ichneumon ferrugator Fabricius (Krombein 1979) * The above mentioned species list represents parasitoids of Hesperiidae and could only potentially be implicated as parasitoids for Urbanus dorantes and Urbanus proteus.

Monarch, Danaus plexippus Linnaeus (Lepidoptera: Nymphalidae)

Danaus plexippus is probably the most charismatic species on earth, which is the result of

its handsome appearance and habit of migrating to Mexico to escape the winter. The location of

the destination in Mexico to which adult D. plexippus in the eastern United States migrated

remained a mystery for many years. In 1977, Kenneth Bruegger discovered that the adults were

heading to Michoacan, Mexico (Urquhart, F., & Urquhart, N. 1976; Brower 1977). The

elevation of the mountains around Michoacan, Mexico is between 10 000 to 11,000 feet and the

forest is dominated by firs. While Monarchs are not considered endangered, the yearly migration

of Monarchs inhabiting localities east of the Rockies to high altitude forests in Mexico is

considered by the International Union for Conservation of Nature as an endangered

phenomenon. The North American Monarch Conservation Plan represents a concerted effort on

the part of scientists, government officials, and laymen to help keep the Monarch migration

intact.

D. plexippusis facing pressures including, but not limited to, (a) habitat destruction, (b) predators (mice and birds), (c) lack of food, and (d) a protozoan called Ophryocystis

elektroschirra (Cech & Tudor 2005).

Distribution: Danaus plexippus are found throughout the state of Florida. Worldwide distribution is quite broad, ranging from western and northern South America to southern

Canada. Following a colonization event in the 19th century, Monarchs were established in

Australia, Micronesia, Madeira, Canary Islands, Spain, Portugal, and other islands throughout the Atlantic and Pacific.

Description: The egg is yellow-white and has an oval shape with a pointed crest. It has very fine ridges rising up from the base and proceeding to the tip of the base.

The larva possesses yellow, black, and white transverse stripes. Four black fleshy filaments protrude out of the caterpillars’ body. One pair comes out of the thorax and the other pair comes out near the rear of the caterpillar. Black strips are located on the white head of the caterpillar.

The pupa is mint green with a black and gold band along the underside of the abdomen.

The head has six pairs of gold spots, which differs from the Queen pupa, Danaus gilippus

(Cramer) (Lepidoptera: Nymphalidae), which has only four pairs at the head. One pair of gold

spots resides near the oculus. Another pair of gold spots resides near the labrum. Two pairs of

gold spots rest between the oculus and the labrum at the head of the pupa, which differentiates

the Monarch from the Queen butterfly. Two final pairs of gold spots occur at the top of the

thorax. The forewing also has a gold dot. The pupa has a black cremaster with a pair of black

spots at the top of the anal area.

The adult wingspan ranges from 8 cm to 11 cm. The upper side of the Monarch butterfly is

chiefly orange with black veins and borders. The black borders and the tips of the forewings

have white spots. The coloration of the underside is chiefly white cream orange on the lower

hind wings and a more pale orange on the forewings. Like the upper side, black covers the veins

and forms the borders of the wings. White spots intersperse the black borders on both the lower

side of the hind wings and fore wings. Males have a small patch of androconial scales on the

hind wings on vein Cu2. The patch is visible on both the upperside and underside of the wings.

Males are slightly larger than the females. In males, hair pencils are present at the tip of the

abdomen. Females lack the small patch of androconial scales on the hind wings on vein Cu2.

They also possess a greater amount of brown scales on the orange patches of the wings and more

black scales along the veins. This makes the female appear to have larger veins and a duller

complexion (Minno, M.C. & Minno, M. 1999).

Biology: Female D. plexippus oviposits a single egg on the host plant. Host plants are from

the family Asclepiadaceae and include

• Sarcostemma clausum; • Morrenia odorata;

• Asclepias curassavica; • Asclepiascurtissii; • Asclepiashumistrata; • Asclepiasincarnata; • Asclepiaslongifolia; • Asclepiasperennis; • Asclepiastomentosa; and • Asclepiastuberosarolfsii (Minno, M. C., & Minno, M. 1999).

The larvae are surface feeders on their host. Later instar larvae will prey upon the earlier

instar larvae and eggs of their own species (Minno, M. C. & Minno, M. 1999).

D. plexippus are multivoltine and in Florida, they are more visible during the spring

(March to May) and fall (August to December), and there is a resident population of Monarchs that are visible in Central and Southern Florida year-round (Minno, M. C. & Minno, M. 1999).

D. plexippus are slow deliberate fliers that cruise at an altitude of 1 to 2 meters, but during migration, they reach higher altitudes between 600 and 1250 meters. Scientists pursuing

Monarchs during this migration period noted that the D. plexippus seemed to be taking advantage of thermals (Gibo 1981). D. plexippus have been reported to be overwintering in

Apalachicola ,Florida as early as 1875-1876 (Kimball 1965). During the 1950’s and 60’s the

original 1875-1876 overwintering sites of D. plexippus and many additional overwintering sites were documented. The new sites included

• Cedar Key; • Lighthouse point; • East point; • St. Joseph’s Key; • Springfeild; • Alligator Point (Wakulla county); • The everglades; • Miami; and • Dry Tortugas, FL (Kindall 1965).

Natural Enemies: While many birds avoid the D. plexippus adult, some birds such as

Orioles, Jays, and Grosbeaks have developed the ability to eat adults (Calvert et al.1979). Several

species of mice are another major predator of the overwintering adults (Brower et al.1985).

Up to ten parasitoids of Lespesia archippivora (Riley) (Diptera: Tachinidae) have been

found inside a D. plexippus larva. It was found that the later the instar the more likely more than

one L. archippivora would be recoverved (Oberhauser et al.2007). The average incidence of

parasitism of D. plexippus larva by L. archippivora was 11% (Oberhauser et al.2007). The

incidence of parasitism by Sturmia convergens Wiedemann (Diptera:Tachinidae)and Paradrino

laevicula (Mesnil) (Diptera: Tachinidae) in Australia varied from 11% to 100% (Zalucki 1981).

Hymenoptera Chalcidoidea Pteromalidae Pteromalus cassotis Walker (Krombein 1979) Perilampidae Perilampus hyalinus (Say) (Oberhauser & Solensky 2004) Trichogrammatidae Trichogramma pretiosum Riley (Querino et al. 2002) Trigonaloidea Trigonalidae Taeniogonalos raymenti Carmean & Kimsey (Clarke & Zalucki 2001) Diptera Oestroidea Tachinidae Exorista mella (Walker) Lespesia schizurae (Townsend) (Arnaud 1978) *Lespesia archippivora (Riley) (Arnaud 1978; Oberhauser et al.2007) **Paradrino laevicula (Mesnil) **Sturmia convergens Wiedemann (Cantrell 1986) *possibly exhibiting superparatism (Oberhauser et al.2007) ** Australian spp. (Cantrell 1986)

Fall Webworm, Hyphantria cunea Drury (Lepidoptera: Arctiidae)

The fall webworm, Hyphantria cunea, is an invader to the United States, Japan, China, and Korea (Nordin & O’Canna 1985; Yang & Zhang 2007). The gregarious larvae form large tents around the host. Currently, the taxonomic status of Hyphantria cunea as a single species remains intact. The variations in phenotypes and behavior have resulted into some discussion concerning the possibility that a sympatric speciation has occurred (Takeda 2005). In Japan,

DNA barcoding of the mitochondria suggests two species are occurring sympatrically along with numerous behavioral and morphological differences between the two forms (Gomi et al. 2004)

Distribution: The Hyphantria cunea’s northern range limit occurs at the latitude of 50-55

° where there are less than 200 days of 42° F (Morris 1963). The single voltine and chiefly black headed and dark bodied larva occurs above the latitude 40°. Whereas the multivoltine cycles persist with both red and black headed larvae and the bodies of the larva are generally lighter green (Wagner et al. 1998). In the United States, the southern limit of its range is Mexico.

Hyphantria cunea has been introduced from the United States to Europe and Asia and has expanded its range so as to be considered to inhabit all suitable habitats of the holarctic region

(Worth 1994).

Description: The egg mass of Hyphantria cunea is almost iridescent green in color. The mean egg mass is between 400 to1000 (Ito et al. 1969).

The larva is hairy and has a lime green body with black spots. Farther north, the larvae are more dark black or brown than lime green and black spotted (Morris 1963). The head capsules are found in two colors red or black (Takeda 2005). Larvae form a large tent structure around the foliage they are consuming. The red morph larva stays in the shelter of the tent throughout the

larval stage, while the black morph leaves the tent after the fifth instar (Ito & Warren 1973;

Takeda 2005). There are five grades of larva color according to Masaki and Ito (1977).

Grade 1- body color yellow, setae white with black spots;

Grade 2- body color a mosaic of yellow and brown, setae white with black spots;

Grade 3- dorsal body gray-black, lateral body yellow, setae white;

Grade 4- body color solid gray to black, lateral body with black gray, and yellow spots , setae white, black turbercules; and

Grade 5- dorsal body is uniformly black, lateral body black or dark gray, setae black

The pupa of Hyphantria cunea is covered with a cocoon that many times contains fragments of leftover foliage. The pupa is brown.

The adult female is immaculate from either the red or black morphs (Takeda 2005). The adult males on the other hand can have spotted or immaculate wings. The red headed larva never produces spotted males, while the black headed males may produce spotted males (Takeda

2005).

Biology: There are definite geographic regions where black headed larvae dominate

(northern climes, e.g. Nova Scotia), red larvae dominate (southern climes, e.g. Florida), and regions where mosaics form (in between the southern and northern climes, e.g. South Dakota)

(Ito & Hattori 1973). The adaption of the black form to emerge prior to the red form is, possibly, caused by the red form’s superior ability to survive in the wild, which could stem from differences in (a) net (b) mating style, (c) time of eating, (d) length of stay in the tent, and (e) inception of diapause (Masaki & Ito 1977; Takeda 2005).

Host Plants: The wide range of the Hyphanea cunea is due to the larvae being the consummate generalist. There is no other insect on the planet that has been recorded eating more plant species than Hyphanea cunea (Worth 1994). The ability to successfully generalize and

consume a wide variety of hosts that may even possess toxic materials is a hallmark of the family

Arctiidae (Krasnoff & Dussourd 1989).

Economic Importance: Hyphanea cunea has caused the destruction of many ornamental

trees and forests throughout the holarctic regions of the world (Franz 1961; Yang & Zhang

2007). It has notably been implicated as a pest of sericulture, because of its preference for

mulberry leaves (Nordin & O’Canna 1985).

Natural Enemies: From the egg to the first larval instar there is approximately 50 to 60 %

mortality, but predation of Hyphanea cunea is most severe from the fourth instar to the pupa

with between 98 and 99 % mortality (Masaki & Ito 1977). The high mortality stems from bird

predation see Table 3-1. Some of the major hymenopteran predators are Polistes annularis

(Linnaeus) (Hymenoptera: Vespidae) and Polistes fuscatus (Hymenoptera: Vespidae) (Krombein

et al. 1979).

Parasitoids:Parasitoids found infecting Hyphanea cunea are

Sinophorus validus (Cresson) (Hymenoptera: Ichneumonidae); Meteorus hyphantriae Riley (Hymenoptera: Braconidae); Apanteles hyphantriae Riley (Hymenoptera: Braconidae); Elachertus hyphantriae Crawford (Hymenoptera: Eulophidae); and Mericia ampelus (Walker) (Diptera: Tachinidae)

which affected the black and red race larvae with an incidence of parasitism at 34% and 39%

respectively (Nordin 1972).

Hymenoptera Ichneumonoidea Braconidae Aleiodes sanctihyacinthi (Provancher) Apanteles diacrisiae Gahan Apanteles hyphantriae Riley Microplitis hyphantriae Ashmead Meteorus bakeri Cook and Davis Meteorus hyphantriae Riley (Krombein et al. 1979)

***Rogas malacosomatos Mason (Mason 1979) Chalcidae *Brachymeria obscurata (Walker) (Ito & Yamada 1968) Ichneumonidae Itoplectis inquisitor Say Vulgichneumon brevicinctor (Say) Ichneumon navus Say Therion morio (Fabricius) Therion sassacus Vier Sinophorus validus (Cresson) Hyposoter fugitivus (Say) Hyposoter rivalis (Cresson) (Krombein et al. 1979) Hyposoter pilosulus Prov Campoplex validus Cress. (Tadic 1977) Enicospilus glabratus Say Casinaria genuina (Norton) Casinaria limenitidis Howard (Krombein et al. 1979) *Coccygomimus disparis (Viereck) (Ito & Yamada 1968) Pimpla turionellae Linnaeus (Arthur & Wylie 1959)

Pteromalidae Dibrachys cavus, (Walker) Eulophidae Elachertus hyphantriae Crawford Syntomosphyrum esurus (Riley) (Krombein et al. 1979) **Chouioia cunea Yang (Yang 2007) Torymidae *Monodontomerus minor (Ratzeburg) (Ito & Yamada 1968) Trichogrammatidae Trichogrammadendrolimi Matsumura (Kato et al. 1951) Diptera Oestroidea Tachinidae Lespesia aletiae (Riley) Chetogena claripennis (Macquart)

Chetogena scutellaris (Van der Wulp) Hyphantrophaga hyphantriae (Townsend) Blondelia hyphantriae (Tothill) (Arnaud 1978) *Exorista japonica Townsend *Zanillia libatrix Panz. *Pales pavida Meigen (Ito & Yamada 1968) Mericia ampelus (Walker) (Tadic 1977) Lespesia frenchii (Williston) (Arnaud 1978; Tadic 1977)

*Japan (Ito & Yamada 1968) **China (Yang 2007) *** parasitoid of Malacosoma spp. (Mason 1979)

Eastern tent caterpillar, Malacosoma americanum (Fabricius) (Lepidoptera: Lasiocampidae)

The eastern tent caterpillar, Malacosoma americanum, is widespread in the eastern part of north America (Stehr & Cook 1968). Larvae of M. americanum emerge from the egg diapause,

due to environmental changes and begin feeding (Fitzgerald 1995).

Distribution: M. americanum is distributed along the eastern part of the United States and

the southeastern part of Canada (Stehr & Cook 1968).

Description: The eggs are laid as a group of 200 to 300 (Stehr & Cook 1968). Eggs are

dark brown.

The larva is partly dark brown and golden yellow. The larvae possess nonpigmental

coloration. The dorsal white stripe and the lateral blue stripe on the larval body are caused

arrangement of the microtubercles (Stehr & Cook 1968). The lateral blue stripes with yellow

borders, on the larval body, are broken up by white and black patches.

The pupa is brown and is surrounded by an off white cocoon.

The adult is brown to reddish brown and hairy. The forewings possess two vertical white stripes. The female has less distinctive white stripes on the forewing and thinner antennae. The males are smaller in size than the females.

Biology: After the oviposition of the eggs by the female, the eggs are very quick to complete embryogenesis, but do not come out of the egg until the next spring (Fitzgerald 1995).

The tents constructed by the larvae are designed according to the orientation of the light source.

In a series of experiments, altering the light source produced a corresponding alteration in the location for the entrance to the larval tent (Fitzgerald & Willer 1983).

Feeding occurs outside the larval tent, which requires the larva to forage. During foraging a silk line is made along the route taken. When food is discovered the silken trail is reinforced with more silk and the pheromone 5b-cholestane-3-one (Fitzgerald 1995). Synthetic duplicates of this pheromone are attractive or even more attractive to larva (Fitzgerald 1995).

Natural Enemies: Larval predation is very high in M. americanum. Its many predators include numerous bird species, certain Hymenoptera, Hemiptera, Coleoptera as well as a host of parasitoids.

After the eggs are laid and sometimes after the eggs have undergone embryogensis, an egg parasitoid oviposits in the egg (Maple 1937). The timing of egg parasitoid eclosion is dependent on whether they are a generalist or a specialist. Specialist egg parasitoids emerge after the larvae have emerged (85 to 115 days following larval emergence (Liu 1926)) and have begun feeding on the host plant, while generalists usually emerge prior to larval emergence

(Fitzgerald 1995).

Hymenoptera Ichneumonoidea Ichneumonidae Hyposoter fugitivus (Say)

(Krombein et al.1979) Labrorychus analis (Say) Gellus tenellus (Say) (Dethier 1980) Enicospilus cushmani Gauld (Gauld 1988) Coccygomimus disparis (Viereck) (Schaefer et al. 1989) Itoplectis conquisitor (Say) (Witter & Kulman 1972) Torymidae Monodontomerus minor (Ratzeburg) (Krombein et al. 1979) Eulophidae Tetrastichus malacosomae Girault Tetrastichus silvaticus Gahan Tetrastichus sp. Braconidae Phobocampa clisiocampae Weed Therion sp. (Dethier 1980)

Platygastroidea Scelionidae Telenomus clisiocampae Riley Chalcidoidea Encrytidae Oencyrtus clisiocampae Ashmead Oencyrtus sp. Aphelinidae Ablerus clisiocampae (Ashmead) Ablerus sp. Eupelmidae Anastatus sp. Trichogrammatidae Trichogramma evanescens West Trichogramma minutum Riley (Fitzgerald 1995) Chalcididae Brachymeria ovata (Say) (Dethier 1980) Diptera Oestroidea Tachinidae Carcelia laxifrons Villeneuve (introduced) Lespesia aletiae (Riley)

Lespesia datanarum (Townsend) Lespesia schizurae (Townsend) Chetogena claripennis (Macquart) Chetogena edwardsii (Williston) Chetogena lophyri (Townsend) Blepharipa pratensis (Meigen)(introduced) Archytas lateralis (Macquart) (Arnaud 1978) *Compsilura concinnata (Meigen) Leschenaultia exul (Townsend) (Fitzgerald 1995) Euphororocera tachinomoides (Townsend) Hyphantrophaga hyphantriae (Townsend) Achaetonera sp. (Dethier 1980) Lespesia archippivora (Williston) Lespesia frenchii (Williston) Exorista mella (Walker) (Arnaud 1978; Dethier 1980) Sarcophagidae **Sarcophaga aldrichi Parker (Hodson 1939) *Obligatory to Malacosoma americanum **Facultative to Malacosoma americanum

Parasitoid Checklists for Six Additional Lepidoptera Hosts Collected During Survey

Tawny emperor, Asterocampa clyton (Boisduval & Leconte) (Lepidoptera: Nymphalidae)

The gregarious behavior of this species gives rise to high incidence of parasitism (Cech &

Tudor 2005).

Hymenoptera Platygastroidea Scelionidae (Friedlander 1985) Diptera Oestroidea Tachinidae Chetogena claripennis (Macquart) (Arnaud 1978)

Polydamas Swallowtail, Battus polydamas lucayus (Rothschild & Jordan) (Lepidoptera: Papilionidae)

Members of the genus Battus seem to be protected from parasitoid attack by the

reservation of chemicals from their host Aristolochia spp. (Sime 2002)

Mourning Cloak, Nymphalis antiopa (Linnaeus) Lepidoptera: Nymphalidae

Hymenoptera Chalcidoidea Pteromalidae Pteromalus puparum Linnaeus (Krombein et al. 1979) Diptera Oestroidea Tachinidae Lespesia aletiae (Riley) Lespesia archippivora (Riley) Lespesia dubia (Williston) Lespesia frenchii Williston Chetogena claripennis (Macquart) Chetogena edwardsii (Williston) Exorista mella (Walker) Hyphantrophaga blanda (Osten Sacken) Hemisturmia parva (Bigot) Winthemia sinuate Reinhard (Arnaud 1978) Cecropia silkmoth, Hyalophora cecropia (Linnaeus) (Lepidoptera: Saturniidae)

Diptera Oestroidea Tachinidae Lespesia datanarum (Townsend) Lespesia frenchii Williston Chetogena claripennis (Macquart) Winthemia datanae (Townsend) (Arnaud 1978) Compsilura concinnata (Meigen) (Boettner et al.2000) Hymenoptera Ichneumonoidea Ichneumonidae Theronia atalantae fulvescens (Cresson) Gambrus extrematis (Cresson) Enicospilus americanus (Christ) (Krombein et al 1979)

Ephialtes capulifera (Kriechbaumer) Theronia atalantae fulvescens (Cresson) Gnotus sp. Gambrus polyphemi H.Townes Agrypon illinois Dasch Braconidae Cotesia sp. Chalcidoidea Torymidae Monodontomerus minor (Ratzeburg) Perilampidae Perilampus hyalinus (Say) Pteromalidae Psychophagus omnivorus (Walker) Dibrachyscavus (Walker) Chalcididae Conura maria (Riley) Eulophidae Dimmockia incongrua (Ashmead) Cirrospilus inimicus Gahan Pediobius tarsalis (Ashmead) Winthemia cecropia (Riley) Winthemia datanae (Townsend) Winthemia leucanae (Kirkpatrick) Winthemia militaris (Walsh) Winthemia quadripustulata (Fabricius) Eusisyropa virilis (Aldrich & Webber) Lespesia sp., archippivora complex Lespesia frenchii (Williston) Lespesia samiae (Webber) (Piegler 1994)

Brazilian Skipper, Calpodes ethlius (Stoll) (Lepidoptera: Hesperiidae)

Hymenoptera Chalcidoidea Encyrtidae *Ooencyrtus calpodicus Noyes Chalcididae Brachymeria incerta (Cresson) Trichogrammatidae Xenufens ruskini Girault (Krombein et al.1979) Trichogramma minutum Riley (Moore 1928). Diptera Oestroidea

Tachinidae Hyphantrophaga blanda (Osten Sacken) Lixophaga diatraeae Townsend (Arnaud 1978) *Caribbean sp.

White-Marked Tussock Moth, Orgyia leucostigma (JE Smith) (Lepidoptera: Noctuidae)

Hymenoptera Ichnemonoidea Ichneumonidae Itoplectis inquisitor Say Pimpla annulipes Say Gelis insolitus (Howard) Adiastola americana Howard (Howard 1897) Apanteles acronyctae (Riley) Apanteles diacrisiae Gahan Apanteles hyphantriae Riley Bracon xanthonotus Ashmead Iseropus coelebs (Walsh) Theronia atlanta fulvescens (Cresson) Gelis insolitus (Howard) Canadensis burkei (Viereck) (Krombein et al.1979) Hyposoter spp. (Guzo & Stoltz 1985) Braconidae Meteorus autographae Muesebeck (Krombein et al.1979) Cotesia melanoscela (Ratzeburg) (Guzo & Stoltz 1985) Cotesia delicatus (Howard) Meteorus hyphantriae Riley Meteorus communis (Cresson) (Howard 1897) Chalcidoidea Pteromalidae Psychophagus omnivorus (Walker) Tritneptis hemerocampae Viereck Eulophidae Syntomosphyrum esurus (Riley) Syntomosphyrum orgyiazle Burks (Krombein et al. 1979) Dipter Oesteroidea Tachinidae

Aphantorhaphopsis samarensis (Villeneuve) (introduced)** (Fuester et al.2001) Chetogena claripennis (Macquart) Exorista griseomicans Van der Wulp Nilea lobeliae (Coquillett) Lespesia aletiae (Riley) Lespesia frenchii Williston Exorista mella (Walker) Winthemia 4-pustulata Fabricius Amorphota orgyiae Howard (Howard 1897) Bessa selecta (Meigen) Chetogena edwardsii (Williston) Chetogena floridensis (Townsend) Hyphantrophaga hyphantriae (Townsend) Patelloa leucaniae (Coquillett) Winthemia datanae (Townsend) (Arnaud 1978)

Survey Protocol

Collecting Localities

In the fall 2009, Lepidoptera caterpillars of species listed above were collected thoughout

Alachua county (Florida). In 2010, Broward and Miami Dade counties were included in the survey. The collecting sites were each grouped into ~19 mi2blocks. When more than one site was

present in a block, the sites were clumped together. Table 3-1 shows the locality data (GPS

readings) for the collecting sites. Figure 3-1 shows the locations for collection sites in the vicinities of Gainesville, Alachua County. Figure 3-2 shows the locations for collection sites in

Miami, FL.

Collecting and Rearing

The methods for collecting larvae were influenced by the work of Daniel Janzen and his

team of parataxonomists in Costa Rica (Janzen & Hallwachs 2009). Janzen’s team begins by

searching for Lepidopteran larvae in the field on a variety of hostplants. The discovered

Lepidopteran larvae are collected and reared to adult hood. The reared adults are recorded and sent to experts for identification and DNA barcoding.

Larvae were collected on host plants in the wild. Initially, all larval instars were included into the survey; however, high mortality during rearing led to a revising of the protocol to collecting 3rd through 5th instars only. Upon collection of a larva in the wild, the host plant, date, and location where the caterpillar was found were recorded. The collected caterpillars were taken to the lab and placed into small plastic or Styrofoam cups. The containers holding the caterpillars were labeled with the date and a voucher number. Each voucher number was entered into a MSExcel file that helped track the dates of collection, pupation, eclosion, larva species, host plant information, and the outcome of rearing (presence or absence of parasitism).

Caterpillars in containers were fed on host plant cuttings or artificial diet (Ward’s Stonefly

Heliothis Diet) until the larva pupated or died. When an adult Lepidoptera or parasitoid eclosed from a pupa, it was frozen at 9-18° Celsius and stored until curation could be undertaken.

Specimen Preservation and Identification

Dead adults were curated and placed into a collection box for storage. Voucher specimens of adult tachinid flies were sent to James E. O’Hara at Invertebrate Biodiversity

Agriculture and Agri-Food Canada, Ottawa, Ontario, Canada for identification and deposition of voucher specimens. Adult parasitoids of the families Ichneumonidae and Braconnidae were identified with the aid of specialists at the Department of Plant Industry in Gainesville, FL and

Dr. Andrei Sourakov at the Florida Museum of Natural History. A leg was removed and placed in 85 % alcohol for future DNA-barcoding analysis. Legs of parasitoids were sent with voucher numbers to the Biodiversity Institute of Ontario at the University of Guelph.

Photography

Voucher specimens of adult parasitoids were photographed with Syncroscopy Auto-

Montage digital microscopy system at the Entomology and Nematology Department at the

University of Florida.

Results and Discussion

Over 3,000 larvae were collected between August 2009 and September 2010. To date, 596 parasitoid pupae and adults have been recovered. The list of species of Lepidoptera larvae collected during the survey is found in Table 3-3. Twenty-six morphospecies were identified from the 596 recovered parasitoids. The list of parasitoid species reared from the Lepidoptera larvae can be found in Table 3-4. Figures 3-3 through 3-22 are Auto-Montage Images of the parasitoids listed in Table 3-4.

The highest number of parasitoids was obtained from Urbanus proteus, and with one exception were species of Tachinidae. Hyphantria cunea followed closely behind, as the host for mostly hymenopteran species. The rate of parasitism was quite variable depending on location and species. Danaus plexipus larvae from MSA nursery in Loxahatchee, FL had 70% parasitism

(N= 54). Conversely, D. plexippus larvae from Fairchild Tropical Botanical Gardens in Miami,

FL had a parasitism rate of 0% (N=15). Interestingly, no parasitoids were recovered from some of the species, such as Heliconius charithonia and Battus polydamus (Table 3-3).

Table 3-1. Life Table for Hyphantria cunea at NS1 in the First generation of 1967 (after Ito & Miyahsita 1968).Note: L I, LII, etc. mean the first-instar larva, second-instar larva, etc. x lx dxF dx 100qx Egg 4287 Unhatched 134 3.1 L hatched 4153 Spiders and 746 18.0 unknown L I 3407 Physiological 104 3.1 causes Spiders and 1093 32.1 unknown L II 2210 Physiological 11 0.5 causes Spiders and 322 14.6 unknown L III 1877 Spiders and 463 24.7 unknown L IV 1414 Great tit young 680 48.1 Mainly adult 693 49.0 birds L VII 41 Birds and 29 70.7 Polistes Prepupa 12 Tachinid 3 25.0 parasites Pupa 9 Tachinid 1 11.1 parasites Disease 1 11.1 Adult 7 Total mortality= 4280 99.84%

Table 3-2. Collection localities for survey of parasitoids in Florida. County Florida Location Map GPS coordinates Cod e Miami-Dade Abandoned Plot, G 25°29'23.13"N 80°28'32.26"W Miami-Dade Fairchild Tropical Botanic E 25°40'36.04"N 80°16'18.22"W Gardens Miami-Dade UF Trop. Research Station F 25°30'24.16"N 80°29'52.84"W Miami-Dade Casey's Corner Nursery H 25°28'13.41"N 80°30'35.01"W Broward South Florida Water A 26° 5'23.07"N 80°25'39.37"W Management District Miami-Dade Mack's Fishing Camp C 25°57'46.40"N 80°27'21.01"W Miami-Dade Everglades Holiday Park B 26° 3'39.95"N 80°26'27.48"W Miami-Dade Bill Baggs Cape Florida D 25°40'37.82"N 80° 9'32.02"W SP Alachua NW 83rd St/NW 39th Ave 0 29°41'19.03"N 82°25'48.89"W Alachua SW 23rd St AA 29°37'36.83"N 82°21'24.46"W Alachua I-75/NW 39th Ave M 29°41'16.74"N 82°26'32.07"W Alachua Natural Area T 29°38'3.76"N 82°22'1.77"W Alachua Kanapaho Park KK 29°37'12.63"N 82°25'4.42"W Alachua SE Williston Rd Z 29°37'51.40"N 82°19'17.28"W Alachua NE Waldo Rd/SE 11th St Y 29°39'0.11"N 82°18'41.60"W Alachua SW 13th St/SW 9th Rd U 29°38'27.97"N 82°20'21.12"W Alachua NW 39th Ave Q 29°41'20.61"N 82°24'24.19"W Alachua Stadium by Pool X 29°39'39.43"N 82°18'25.29"W Alachua Apartments near dead end W 29°40'11.59"N 82°18'4.20"W Alachua SW 34th/ SW 63rd Ave JJ 29°35'35.78"N 82°21'36.28"W Alachua Lake Wauberg South GG 29°31'20.96"N 82°18'9.01"W Alachua SW 63rd Ave DD 29°35'39.03"N 82°20'27.47"W Alachua SW 66 PL CC 29°35'28.65"N 82°20'29.19"W Alachua SE 134th Ave HH 29°31'55.31"N 82°19'2.69"W Alachua Lake Wauberg North EE 29°32'0.13"N 82°18'30.61"W Alachua SE 134th Ave (2) FF 29°31'47.99"N 82°18'36.50"W Bradford Shady Oak butterfly farm I 29°51'23.94"N 82°15'16.03"W Alachua Railroad tracks off of 301 K 29°42'50.09"N 82° 8'22.18"W Alachua Bat Conservancy J 29°49'33.24"N 82°20'27.90"W Alachua Long Leaf Pine Plot L 29°41'33.82"N 82°27'35.76"W Alachua NW 98 St/NW 37 Pl N 29°41'14.63"N 82°27'6.37"W Alachua Santa Fe library P 29°40'59.86"N 82°25'52.87"W Alachua NW 23RD St R 29°40'26.48"N 82°24'2.56"W Alachua 34th St S 29°40'53.43"N 82°22'9.98"W Alachua 2nd Ave/SE 6th St V 29°39'1.57"N 82°19'7.99"W Alachua SW 63rd Ave (Farm) BB 29°35'36.51"N 82°21'13.25"W

Figure 3-1. Map of Gainesville with collection sites labled. See Table 3-1 for names of collection sites.

Figure 3-2. Map of Miami, FL with collection sites labeled by letters. See Table 3-1 for names of collection sites.

Table 3-3. Taxa, whose larvae were collected and reared as part of parasitoid study in Florida in 2009-2010. Species Family #of individual larvae Number of collected parasitoids reared Battus polydamas Papilionidae 27 0 Agraulis vanillae Nymphalidae 280 33 Heliconius charithonia Nymphalidae 42 0 Danaus plexippus Nymphalidae 54 13 Asterocampa clyton Nymphalidae 31 2 Nymphalis antiopa Nymphalidae 2 2 Dryas iulia Nymphalidae 1 0 Euptoieta claudia Nymphalidae 2 1 Urbanus proteus Hesperiidae 1,252 341 Urbanus dorantes Hesperiidae 14 1 Calpodes ethlius Hesperiidae 42 0 Hyphantria cunea Arctiidae 444 177 Malacosoma americanum Lasiocampidae 144 10 Hyalophora cecropia Saturniidae 1 1 Antheraea polyphemus Saturniidae 1 0 Orgyia leucostigma Lymantriidae 22 6 Lymantriidae spp. Lymantriidae 26 10

Table 3-4. Diversity of Parasitoids affecting Lepidoptera hosts. Host Species Location Parasitoid Diversity of Parasitoids Urbanus proteus Gainesville, FL Diptera:Tachinidae Chrysotachina sp. 1 Gainesville, FL Diptera:Tachinidae Chrysotachina sp. 2 Miami, FL Hymenoptera: Braconidae Braconid sp. 1 Urbanus dorantes Gainesville, FL Diptera:Tachinidae Tachinid sp. 3 Agraulis vanillae Gainesville, FL Diptra: Tachinidae Tachinid sp. 4 Gainesville, FL Hymenoptera: Chalcidoidae Chalcid sp. 1 Miami, FL Hymenoptera: Chalcid sp. 2 Chalcidoididae Miami, FL Hymenoptera: Chalcid sp. 3 Chalcidoididae Asterocampa clyton Gainesville, FL Diptera: Tachinidae Tachinid sp. 5 Euptoieta claudia Miami, FL Hymenoptera: Ichneumonidae sp.1 Ichneumonidae Celastrina Huntindon Co. Diptera: Tachinidae Tachinid sp. 6 neglectamajor PA Hymenoptera: Campopleginae sp. Ichneumonidae 1 Danaus plexippus Loxahatchee, FL Diptera: Tachinidae Tachinid sp. 7 Nymphalis antiopa Gainesville, FL Diptera: Tachinidae Tachinid sp. 8 Malacosoma Gainesville, FL Diptera: Tachinidae Tachinid sp. 9 americanum Hymenoptera: Campopleginae sp. Ichneumonidae 2 Hyphantria cunea Gainesville, FL Hymenoptera: Mesochorinae sp. 1 Ichneumonidae Gainesville, FL Hymenoptera: Braconidae Braconidae sp. 2 Gainesville, FL Hymenoptera:Chalcidoidea Chalcidae sp. 4 Gainesville, FL Hymenoptera: Chalcidoidea Chalcidae sp. 5 Hyalophora Gainesville, FL Hymenoptera:Chalcidoidea Chalcidae sp. 6 cecropia Orgyia leucostigma Gainesville, FL Diptera: Tachinidae Tachinid sp. 10 Lymantriidae Gainesville, FL Hymenoptera: Chalcidoidea Chalcidae sp. 7 Noctuid sp. 1 Miami, FL Hymenoptera: Braconidae Braconid sp. 3 Miami, FL Hymenoptera: Bracnoidae Braconid sp. 4 Noctuid sp. 2 Miami, FL Hymenoptera: Braconidae Braconid sp. 5

Figure 3-3. Chrysotachina sp. 1, voucher # 833, (Diptera: Tachinidae) a larval parasitoid of the Longtail Skipper Urbanus proteus (Lepidoptera: Hesperiidae).

Figure 3-4. Chrysotachina sp. 2,voucher # 147, (Diptera: Tachinidae)a larval parasitoid of the Longtail Skipper, Urbanus proteus (Lepidoptera: Hesperiidae).

Figure 3-5. Aplomya theclarum,voucher # 1467, (Diptera: Tachinidae)a larval parasitoid of the Appalachian Azure, Celastrina neglectamajor (Lepidoptera: Lycaenidae).

Figure 3-6. Tachinidae sp. 4.voucher # 805, (Diptera: Tachinidae)a larval parasitoid of theGulf Fritillary, Agraulis vanillae (Lepidoptera: Nymphalidae).

Figure 3-7. Tachinidae sp. 5, voucher # 807, (Diptera: Tachinidae) a larval parasitoid of theDorantes Longtail, Urbanus dorantes (Lepidoptera: Hesperiidae).

Figure 3-8. Tachinidae sp. 6, voucher # 1460, (Diptera: Tachinidae) a larval parasitoid of theMonarch, Danaus plexippus (Lepidoptera: Nymphalidae)

Figure 3-9. Tachinidae sp. 7, voucher # 1168, (Diptera: Tachinidae)a larval parasitoid of the: Eastern Tent Caterpillar Moth, Malacosoma americanum (Lepidoptera: Lasiocampidae).

Figure 3-10. Tachinidae sp. 8, voucher # 1148, (Diptera: Tachinidae) a larval parasitoid of host White-marked Tussock Moth, Orgyia leucostigma (Lepidoptera: Lymantriidae).

Figure 3-11. Tachinidae sp. 9,voucher # 1337, (Diptera: Tachinidae)a larval parasitoid of the Mourning Cloak, Nymphalis antiopa (Lepidoptera: Nymphalidae).

Figure 3-12. Tachinidae sp. 10.voucher # 807, (Diptera: Tachinidae) a larval parasitoid of theDorantes Longtail,Urbanus dorantes (Lepidoptera: Hesperiidae),

Figure 3-13. Campopleginae sp. 1,voucher # 3019, (Hymenoptera: Ichneumonidae) a larval parasitoid of the Eastern Tent Caterpillar, Moth, Malacosoma americanum (Lepidoptera: Lasiocampidae)

Figure 3-14. Campopleginae sp. 2,voucher # 1470, (Hymenoptera: Ichneumonidae) a larval parasitoid of theAppalachian Azure, Celastrina neglectamajor (Lepidoptera: Lycaenidae).

Figure 3-15. Chalcidoidea sp. 1, voucher # 1321, (Hymenoptera: Chalcidoidea) a larval parasitoid of theTussock moth (Lepidoptera: Lymantriidae)

Figure 3-16. Mesochorinae sp 1, voucher # 981, (Hymenoptera: Ichneumonidae) a larval parasitoid of the Fall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae).

Figure 3-17. Braconidae sp. 1, voucher # 1046, (Hymenoptera: Braconidae) a larval parasitoid of the Fall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae).

Figure 3-18. Chalcidoidea sp. 1, voucher # 1101, (Hymenoptera: Chalcidoidea) a larval parasitoid of the Fall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae).

Figure 3-19. Chalcidoidea sp. 2, voucher # 1453, (Hymenoptera: Chalcidoidea) an egg parasitoid of the Cecropia silkmoth, Hyalophora cecropia (Lepidoptera: Saturniidae).

Figure 3-20. Chalcidoidea sp. 3, voucher # 1454, (Hymenoptera: Chalcidoidea) a larval parasitoid of theFall Webworm, Hyphantrea cunea (Lepidoptera: Arctiidae).

Figure 3-21. Chalcidoidea sp. 4, voucher # 1510, (Hymenoptera: Chalcidoidea) a larval parasitoid of theGulf Fritillary,Agraulis vanillae (Lepidoptera: Nymphalidae).

Figure 3-22. Chalcidoidea sp. 5, voucher # 2402, (Hymenoptera: Chalcidoidea) a parasitoid of the Gulf Fritillary, Agraulis vanillae (Lepidoptera: Nymphalidae).

CHAPTER 4 DIFFERENCES BETWEEN HOST-PARASITOID RELATIONSHIPS IN URBAN AND RURAL SETTINGS

Urbanization and Parasitoidism

Butterflies have managed to survive in both urban and rural environments. The world is currently undergoing urbanization at an unprecedented rate. As of 2001, 80% of Americans now live near urban centers (USCB 2001). The scale of urbanization threatens to erode the balance of biodiversity and create an environment with vacant niches for invaders to dominate (McKinney

2006). As Figure 4-1 illustrates, in 2000, the amount of land dedicated to urbanization was significantly greater than the amount dedicated to conservation (Stein 2000). Habitat destruction and/or alteration is one of many factors affecting species survival. In the present chapter, the effects produced by the alteration of landscape from rural to urban are examined in a brief literature review. This review of urban-rural relationships will provide another factor in species survival, a background to the question of butterfly population decline in South Florida, and how this question is related to parasitoids. Concerning parasitoids, special attention will be given to the Tachinidae in the order Diptera.

The Effect of Urbanization on Biodiversity

The destruction of habitat by urbanization creates large tracts of habitat that are very similar. Inside these tracts are pockets of unaltered habitat that, in part, resemble the original ecosystem. The creatures that inhabit these small pockets of natural habitat are many times overpowered by “weed-like” species that are present in many urban environments. The “weed-

like” species are able to overpower the native species and become established in the remaining

pockets of natural (rural) habitat, because of a lack of predatory pressure (McKinney 2002). As a

result, the diversity of plants, birds, insects, and mammals in urban centers is less than half the

diversity found in rural areas (Kowarik 1995; Kurta 2002; Denys and Schmidt 1998; McIntyre

2000; Mackin-Rogalska et al. 1998). Sometimes, as in the European Carabid beetles

(Coleoptera:Carabidae), species do not respond to urbanization, and maintain population levels

(structure) similar to those in rural areas (Niemela 2002). Nonnative species tend to have greater population sizes towards the centers of urban areas (Stein 2000).

While the species diversity of insects, birds, mammals, and plants is reduced in urban centers, suburban areas have an increase in diversity. The species diversity of birds, butterflies, mammals, bumblebees, ants, lizards, and plants has been shown in some suburban areas to be greater than the diversity found in rural areas (Racey & Euler 1982; Blair 2001; Pawlikoski &

Pokornieka 1990; Nuhn & Wright 1979; Germaine & Wakeling 2000; Kowarik 1995). Suburban areas that have higher animal diversity than rural areas also tend to have a high diversity of plant species. Urban centers have a greater proportion of man made structures and materials than suburban or rural spaces and thus are lower in biodiversity . The diversity of birds, insects, mammals, amphibians, and reptiles is related to the species diversity and abundance of plants

(Majer 1997; Shugart et al. 1975; Goldstein et al. 1986; Dickman 1987; McIntyre 2000).

Hypotheses for the Effect of Urbanization on Parasitoidism rates

The differences noted above occurring along urban-rural gradients bring into question the impact of urbanization to a given area. Three hypotheses are cited by Hamback et al. (2007) to describe the ecosystems found in modified habitats. First, rates of immigration of species are decreased and extinction rates of species are increased locally as the size of the core habitat is reduced and the size of the edge habitat is increased (Hanski 1999). Second, local growth rates are adversely affected by modified habitats (Summerville & Crist 2004). Third, the habitat specialists are impacted negatively by the increase in the abundance of generalists and matrix specialists that flourish within the smaller core habitat and larger edge habitats. Furthermore, the

increase in numbers of alternative species that are generalists or matrix specialists may cause species diversity to remain high, but modified (Burke & Nol 2000; Ewers & Didham 2008).

The above-mentioned hypotheses are reiterated to give perspective on the effect of habitat modification on species diversity. It is interesting to note that in the literature there is a wide disparity of opinions regarding the effect of urbanization or habitat modification on biodiversity.

Sometimes, the correlation between plot size and biodiversity is positive, while other times the correlation is neutral or negative (Bonier et al.2007; Bowers & Matter 1997; Clergeau et al.

1998; Clergeau et al. 2001; Connor et al. 2000; Didham et al. 1996; Hamback et al. 2007; Kurta et al. 1992; McGeoch & Chown 1997; Niemelä et al. 2002; Raguso & Llorente-Bousquets 1990).

There is an unequal effect of habitat modification on various trophic levels in the ecosystem.

Parasitoids, for instance, are particularly vulnerable to habitat modification (Didham et al. 1996).

Pockets of isolated herbivore host plants surrounded by disturbed habitats have been shown to have a significantly lower incidence of parasitoid abundance (Kahn & Cornell 1989; Kreuss &

Tscharntke 1994). At a different level, parasitoid species abundance has also been noted to vary temporally and spatially (Inclan and Stireman 2011).

Urbanization and habitat destruction or modification can cause alteration of the food webs. Nonnative plants, for example might replace native vegetation. Similarly, herbivorous insects, predatory insects, and parasitoids can begin to consume the available energy or food resources. The evolving urban ecosystem may appear to have a significant amount of species diversity, but the energy flow within the ecosystem that formerly included only native species must now include invasive organisms on all trophic levels (Tylianaskis et al. 2007).

Parasitoid abundance and diversity should decrease towards the urban center and increase in the rural and suburban areas. The level of increase in parasitoid abundance and diversity is

dependent on edge effects and plant species diversity and abundance (Didham et al. 1996; Kahn

& Cornell 1989; Kreuss & Tscharntke 1994).

Objective

The present study attempts to investigate parastioid populations in both urban and rural settings in Alachua County, Broward County, and Miami-Dade County, Florida over the course of one year.

Materials and Methods

Habitat Types

A variety of habitats found in Florida were also represented in our study: at least one and sometimes more than one of the six most common habitats in Florida were present within 19 mi2

block from each of the collecting sitea. The typical Florida habitats include:

Tropical hammock

Tropical hammocks are threatened by urbanization in South Florida. Hammocks range

from Miami-Dade County to Martin County. This tropical habitat contains plants such as

• Florida boxwood, Schaefferia frutescens Jacq. (Celastrales: Celastraceae) • Lignum vitae, Guaiacum sanctum Linnaeus (Zygophyllales: Zygophyllaceae) • Manchineel tree, Hippomane mancinella Linnaeus (Malpighiales: Euphoribiaceae) • Gumbo limbo, Bursera simaruba (Linnaeus) Sarg. (Sapindales: Burseraceae) • Black ironwood, Krugiodendron ferreum (Vahl) Urban (Rosales: Rhamnaceae) and • Inkwood, Exothea paniculata (Juss.) Radlk. (Sapindales: Sapindaceae) (Karim & Main 2009)

Pine flatwood

Pine flatwoods are the most widespread habitat type in Florida. The soil is not well

drained, and the dominant vegetation consists of the following species:

• Longleaf Pine, Pinus palustris Mill. (Pinales:Pinaceae) • Slash Pine, Pinus elliottii Engelm. (Pinales: Pinaceae) • Pawpaw, Asimina reticulata Shuttleworth (Magnoliales: Annonaceae) • Tarflower, Befaria racemosa Vent. (Ericales: Ericaceae) and

• Fetterbush, Lyonia lucida (Lam.) (Ericales: Ericaceae) (Minno & Minno 1999).

Freshwater marshes

Fresh water marshes are wetlands that have no or very few trees. Plants that are present in

this habitat include

• Carolina Willow, Salix caroliniana Michx. (Malpighiales:Salicaceae) • Wax Myrtle, Myrica sp. (Fagales: Myricaceae) • Saltbush, Atriplex sp. (Caryophyllales: Amaranthaceae) • Silverling, Baccharis halimifolia, (Asterales: Asteraceae) • Elderberry, Sambucus canadensis Linnaeus (Dipsacales: Adoxaceae) • Alligator flag, Thalia geniculataLinnaeus(Zingerberales: Marantaceae) (Minno & Minno 1999),

Disturbed habitat

As a result of urbanization and agriculture there are large areas of disturbed habitat. These areas can be of importance to Lepidoptera conservation becasues they are dominated by by many nectar and hostplants for butterflies. Plants that dominate disturbed habitats include

• Florida beggarweed, Bidens alba Linnaeus (Asterales: Asteraceae) • Tick-trefoil, Desmodium spp. (Fabales: Fabaceae) • Passion-flower, Passiflora spp. (Malpighiales: Passifloraceae)

Beaches

The beach is a very harsh habitat that does not have a large variety of Lepidoptera. Plants

that dominate this region include

• Coco Plum, Chrysobalanus icacoLinnaeus (Malpighiales: Chrysobalanaceae) • Beach Croton, Croton punctatusN. von Jacquin(Malpighales: Euphorbiaceae) • Saw Palmetto, Serenoa repens (Bartram) J. K. Small (Arecales: Arecaceae) • Golden Creeper, Ernodia littoralis, ((Small) R. W. Long (Rubiales: Rubiaceae) • Railroad vine, Ipomea pes-caprae(Linnaeus) R. Brown (Solanales: Convolvulaceae) • Passion-flower, Passiflora spp. (Malpighiales: Passifloraceae) (Minno & Minno 1999).

Mangrove

Mangrove habitat consists of trees that flourish around sea level and the high spring tide

mark.Mangrove trees that occur in this habitat include the

• Red mangrove, Rhizophora mangleLinnaeus (Malpighiales: Rhizophoraceae)

• Black mangrove, Avicennia germinans (Linnaeus) Linnaeus (Lamiales: Acanthaceae) and

• White mangrove, Laguncularia racemosa (Linnaeus) C.F. Gaertn. (Myrtales: Combretaceae)

(Rey & Rutledge 2002)

Urban versus Rural Settings

Each of the collection sites possessed one or more of the above mentioned habitat types.

To determine whether the habitats found at the collection sites were in urban or rural settings

satellite imagery was employed via Google Earth. Figure 4-2 illustrates the 216 cm2 divisions.

The satellite images were scaled to 2 cm=3130 ± 2.4 feet. The scale bar was not identical for each ~ 19 mi2 block, because of variation in the zoom settings in Google Earth. Grids were

formed using Photoshop. The following categories were used to determine the nature of each

square found on the grid, which are as follows: Pasture/Agriculture, Roads, Buildings, Wetlands,

and Trees and Shrubs. If a cm2 on the statelite image, visibly possesses none of the above mentioned categories, it was given a colored dot. After all the dots had been assigned, they were

tallied.

Data Analysis

Larvae were collected and reared to pupation according to the steps outlined in the

Meterials and Methods section of Chatper 3. The outcome of rearing of each caterpillar was recorded and databased according to their collection site and species. During the course of the survey, the suburban and rural sites were visited multiple times at different times of the year in

order to collect caterpillars. To perform the data analysis, parasitoidism rates for the caterpillars collected during one season and within a single collecting locality were considered a single data point.

Statistical analysis

The Mann-Whitney, a nonparametric rank sum test was used to compare the percentages of parasitoids obtained from the larvae of Urbanus proteus in pesticide settings and non- pesticide settings.

The equation for the Mann-Whitney statistic for the percentage of parasitoids found in urban settings is given below.

 (nu )(nu +1) Uu = (nu )(nr ) +   − Tu  2 

The equation for the Mann-Whitney statistic for the percentage of parasitoids found in urban settings is given below.

 (nr )(nr +1) Ur = (nu )(nr ) +   − Tr  2 

The standard errors and means for the urban and rural samples are calculated from the data in

Table 4-3.

Standard Error Equation for urban settings (u=urban)

S S = f X u n

For the remainder of species that had a total sample size n<7, confidence intervals were used to compare the of percentage of parasitoidsobtained in urban and rural settings in

Gainesville, FL and Miami, FL. The small sample sizes prevented an accurate assessment from a normality test.

Confidence Interval Equation

µ = ± × X sX t To complete the Confidence Interval calculation we use degrees of freedom (n-1=t) with a level of confidence of .95.

Confidence Interval Equation for urban settings

µ = u ± × X sX u t

Standard Error Equation for rural settings (r=rural)

S S = f X r n

The non-pesticide collection settings possessed number of samples from the rural collection

settings. Therefore, a different Confidence Interval was used. The degrees of freedom (n-1=t)

with a level of confidence of .95.

Confidence Interval Equation for rural settings

µ = r ± × X sX r t

Results

Urban and Rural Settings

Total tallies of the grid were made into 14 blocks of ~19 mi2. Figure 4-3 illustrates that rural and urban habitats have significant differences between the incidence of streets and buildings. The differences between urban and rural habitat according to the above mentioned criterion would have been more significant without including Bill Baggs Cape Florida State

Park, which possessed a large amount of water cells in the ~ 19 mi2 block that gave an abnormal

urban setting. Figure 4-3 also shows that there is no significant difference between the incidence

of trees and shrubs in urban and rural settings. Figures 4-4 through Figures 4-17 graphically

depict the percentage of cells (216 total) possessing Pasture/Agriculture, Roads, Buildings,

Wetlands, and Trees and Shrubs.Table 4-1illustrates how the graphs were quantified. Table 4-2 outlines the total results for the urban and rural settings and also includes the habitat type(s) found ineachblock. Setting identification using satellite images yielded three rural sites and five urban sites in Gainesville, FL. In Miami, FL, setting identification using satellite images yielded five urban sites and one rural site. Bill Baggs Cape Florida State Park falsely appears to be a rural site (see Figure 4-17 and explanation above), but is misleading because of being an island and having less possible land.

The key differences between the urban and rural habitat were the percentage of buildings and roads found in the grid. In Gainesville, rural settings averaged 27.2% and 22.4% for presence of roads and buildings, respectively. Conversely, urban habitat averaged 69.7 % and

66.2% for presence of roads and buildings, respectively. In Miami, only one rural location proved to yield the proper type of larvae for comparison. Urban locations in Miami, averaged

86.6% and 66.4 % for presence of roads and buildings, respectively.

While there were key differences in the presence of buildings and roads, none of the sites was lacking in trees and shrubbery. Wetlands habitat was not the dominant habitat in any of the sites: Gainesville sites had a median of only 2.3 % and Miami sites had a mean of 11.7 % of water The South Florida Water Management Site had a 46% presence of wetlands.

Pasture/Agriculture prevalence at the sites was more prevalent in Miami (Mean 29.3 %) than

Gainesville (Mean 10.875 %). In Miami, FL, the sites in Homestead, FL were had a high prevalence of agricultural sites with a mean of 58.7%.

Parasitoidism Ratios in Rural Versus Urban Habitats

Gainesville, FL

The number of parasitoids affecting Lepidoptera hosts was significantly different for several species of Lepidoptera in Gainesville between urban and rural habitat. The most

significant result was found in Urbanus proteus, which showed a much higher rate of parasitism in urban settings, see Figure 4-21. After conducting the Mann-Whitney test (P<0.05) for data from Urbanus proteus, the null-hypothesis was rejected that the two populations are identical and the alternative hypothesis that the percentage of parasitioidism are different was accepted.

All of the parasitoids that affected U. proteus were Tachinids. Like Urbanus proteus, Agraulis vanillae tended toward more parasitism in urban settings, but the difference was reliant on only three sites and a smaller sample size, see Figure 4-18.Hyphantrea cunea tended to show higher parasitism in rural settings during the month of July by tachinids, but the difference was like,

Agraulis vanillae, due to a small number sample locations. There was an interesting difference in parasitoids affecting Hyphantrea cunea in one rural sight between April and June in 2010, see

Figure 4-20. Further study of the phenology of parasitoids affecting Hyphantrea cunea throughout the year could prove interesting.

Miami, FL

In Miami, poor sample sizes and only one rural site resulted in asignificant difference between urban and rural sites, see Figure 4-22 and Table 4-3. A. vanillaewere parasitized by

Hymenoptera:Chalcidae in Miami, FL while in Gainesville, FL they were parasitzed by

Diptera:Tachinidae. The small sample size was a limiting factor in the ability to analyze the data form A. vanillae.

Discussion

Frank and McCoy (2007) only considered introduced biological conrol agents that had become established in Florida. Hawkins et al. (1999) noted that established biological control agenst are not as detrimental to nontarget species as non-established biological control agents.

The ten biological control agents considered to be potentially harmful in Florida, have not been documented up to this point as adversely affecting nontarget species. More research is needed to

assess the impact these biological control agents are having on nontarget species in Florida.

Regarding Lepidoptera, two families of leaf miners (Gracillariidae and Lyonetiidae) are particularly at risk of biological control agents (Frank and McCoy 2007).

Higher incidence of parasitoids in urban areas supports the hypothesis that altered habitats may have high species diversity and that habitat modification affects the integrity of preexisting food webs (Tylianakis et al. 2007). In Ecuador, Tylianakis et al. (2007) found that species richness remained high in both undisturbed and disturbed habitats, however the food web structure was significantly different.

Mysteriously, aggregations, of parasitoids are known to occur in locations where their host is even absent (Hassell 1984). The insect herbivore population is not necessarily correlated to the presence of suitable host plants and habitat; parasitoids are thought to be responsible for this phenomenon. For example, Orgyia vetusta Boisduval (Lepidoptera: Lymantriidae) was found in fragmentary spaces throughout seemingly suitable habitat, which was explained by their interaction with predators and parasitoids (Maron & Harrison 1997).

The effect of parasitoids on insect populations has been the object of several metadata analyses. Hawkins states that the susceptibility hypothesis is the most likely method to explain the host deaths caused by parasitoids (1994). The susceptibility hypothesis states that feeding niches/refuges are the key factors affecting the relationship between the host and parasitoid.

Feeding niches are the host plants that herbivores feed on and the predators that prey upon the herbivores. The refuges provide protection for the hosts and serve as a barrier for the parasitoids.

The combination of these two factors affects the numbers of parasitoids and their hosts in an ecosystem (Hawkins 1994; Hawkins et al. 1993). Models that accurately predict the effect of

parasitoid populations on non-target host species are important in making decisions regarding the introduction of biological control agents (Frank & McCoy 2007; Roitberg 2000).

Figure 4-1. Land usage in the United States from 1960 to 2000 (after Stein et al. 2000).

Figure 4-2. Habitat quantification via Satellite Images. August 1, 2010, Google Earth, Scale: 3130 ft. per 2 cm, 12 cm x 18 cm (total area 216 cm2) Total Area: 18.98 mi2

Table 4-1. Habitat quantification for YMCA Road Gainesville, FL Scale: 3128 ft per 2 cm. # cm2 % mi2 Pasture/Agriculture 12 5.6 1.0 Roads 65 30.0 5.7 Buildings 3 1.4 0.3 Wetlands 11 5.1 1.0 Trees and Shrubs 207 95.8 18.2 Total 216 100 19.0

Table 4-2. Description of Habitat types. Site name Habitat Type(s) Setting % p % s % b % w % t YMCA Pine Flatland, Freshwater Marsh, r Disturbed Habitat 6.0 30.0 1.4 5.1 96.0 SW 63rd Pine Flatland, Disturbed Habitat r 11.0 26.4 11.0 0 100.0 Shell Pine Flatland, Disturbed Habitat r 37.5 25.5 11.0 0 100.0 NW 98 St Pine Flatland, Freshwater Marsh, u 14.0 54.2 38.4 1.4 100.0 Disturbed Habitat Kanapaho Park Pine Flatland, Freshwater Marsh, u 0 55.0 49.1 10.2 100.0 Disturbed Habitat NATL Pine Flatland, Freshwater Marsh, u 7.9 84.0 82.4 3.7 96.8 Disturbed Habitat Santa Fe Pine Flatland, Disturbed Habitat u 0 58.8 66.2 0 100.0 Stadium by Pool Gainesville, Pine Flatland, Disturbed Habitat u 0 96.3 94.9 0 99.5 FL Bill Baggs Cape Florida State Pine Flatland, Disturbed Habitat, u 0 31.5 18.5 3.2 32.8 Park* Beaches, Mangrove, Tropical Hammock Casey's Corner Nursery, Pine Flatland, Disturbed Habitat u 54.2 88.4 54.2 0 78.2 Homestead, FL UF Tropical Research Station Pine Flatland, Disturbed Habitat u 91.2 93.1 51.9 0 65.3 Abandoned Plot, Homstead, Pine Flatland, Disturbed Habitat u 30.6 96.8 94.0 0 71.8 FL Fairchild Botanical and Disturbed Habitat, Mangrove, u 0 68.1 65.3 20.4 82.9 Tropical Gardens, Tropical Hammock South Floida Water Pine Flatland, Disturbed Habitat, r 0 32.9 6.0 46.8 100 Management Freshwater Marshes

Note: urban=u, rural=r, pasture/agriculture=p, roads=s, buildings=b, wetlands=w, and trees and shrubs =t. * Bill Baggs Cape Florida State Park is located on a peninsula, which affects the ratios

Table 4-3. Sample sizes and parasitoid percentages (parasitoids/total adults eclosed) for each collection site in Gainesville and Miami, FL Lepidoptera and Sample Size Quadrat Number 1 2 3 4 5 Location Urbanus proteus n=306 Urban (parasitoid/total .301 ,214 ,620 .666 Gainesville, FL ecalosed)

n=120 Rural (parasitoid/total eclosed .01086 0 0 ) Agraulis vanillae n=63 Urban (parasitoid/total .41 ,32 Gainesville, FL eclosed)

n-16 Rural (parasitoid/total eclosed) 0

Hyphantrea cunea n=128 Urban (parasitoid/total .187 .696 Gainesville, FL eclosed)

n=294 Rural (parasitoid/total eclosed) 0.056 .0476

Agraulis vanillae n=77 Urban (parasitoid/total .143 .304 .143 .2 0 Miami, FL eclosed)

n=2 Rural (parasitoid/total eclosed) 0

100

character

of cells possessing a certain a certain possessing of cells

Percentage (%) Percentage r

/Shrubs /Shrubs Trees Rural Trees/Shrubs Urban Streets Rural Streets Urban Buildin Rural Buildings Urban

Figure 4-3. Comparison and contrast of the range of percentages of three characters (streets, buildings, and trees/shrubs) found in thirteen collection areas.