December 2013 IDENTIFIER NOTES OF INTEREST NEWSLETTER Page 1

The IDF plan addresses potential issues What is ID Facilitation? of delay in the PPQ pest identification system by: DA’s office charged NIS and Field -Op Message from Joe Cavey, erations (FO) managers with addressing 1. Investigating general and NIS Assistant Director the timeliness issues of pest identifica- specific (to location) causes of delay to tion processing. The resulting IDF plan pest interception processing. (Note: n December 6, 2013, a message was approved and supported with the pest interception processing is defined as Ofrom the PPQ Deputy Adminis- addition of 30 new positions. In this the period from detection of the pest to trator announced a plan to hire 30 pest article, I want to briefly introduce you the time the action decision is delivered identification positions beginning in to aspects of IDF other than staffing. to the broker, to include CBP and PPQ January 2014. The new positions will actions.) continued on page 8 include PPQ identifiers, national taxo- nomic specialists and positions under a Good Progress Made by the PPQ Molecular new GS-11 Plant Health Safeguarding Diagnostics Task Force by Joel Floyd, NIS Domestic Specialist/Pest Identification position Diagnostics Coordinator description. Discussions are under way hanks to the with NAAE, affected SPHD’s and CBP Thard work of on what is the most significant employ- the PPQ Molecular ment incident affecting the pest identifi- Diagnostics Task cation function in 40 or more years. As Force (MDTF), we Mr. El-Lissey stated in his message, we are closer to a goal of will announce the openings and loca- deploying molecular tions in January. We will also share our diagnostics for pest hiring plans, as all positions will not be identification at announced and opened at once. ports of entry. In the coming year, there This employment initiative is part of are three pilot projects planned to take CONTENTS a larger effort named Identification place at Plant Inspections Stations (PIS) page Facilitation (IDF). The IDF project around the country. More on that later, What is ID Facilitation? . . . 1 was developed to respond to increased but first let’s catch everyone up on this Molecular Diagnostics . . . . 1 demands on PPQ’s pest identification significant effort. Mary Joyce Burns...... 2 system, especially regarding timeliness While discussions began in early 2011, NIS Urgent Policy Guide . . 3 of processing cargo interceptions for Mary Palm and Joe Cavey began in New Nikon Camera ...... 4 identification. These demands result 2012 gathering personnel from various Fruit Fly DNA ...... 6 from increases over the past decade in functions and levels in PPQ including Insect ID Videos ...... 9 cargo volume, numbers of pest intercep- identifiers, PIS supervisors, CPHST image ID...... 10 tions, numbers of pest taxa intercepted, molecular biologists, Field Opera- Woodborer Rearing ...... 12 hours of cargo inspection operation and tions managers, taxonomic specialists, industry expectations. With support Historic Photos ...... 14 of the APHIS Administrator, the PPQ continued on page 7 Page 22 IDENTIFIER IDENTIFIER NOTES NOTES OF INTEREST OF INTEREST NEWSLETTER NEWSLETTER December December 2013 2013 Many Happy Returns: A Big Thanks to Mary Joyce Burns, NIS’ Program Assistant

dentifiers in entomology may have Inoticed more regular and increased numbers of specimens being returned from the ARS Systematic Entomology Laboratory (SEL) in the last year or so. This is thanks to the dedication and diligence of Mary Joyce Burns, our NIS Program Assistant who takes her job very seriously, and enjoys it too.

After the SEL specialists do their iden- tifications / confirmations, the intercep- tions are then routed through the SEL Communications & Taxonomic Services Unit in Beltsville, MD where they are checked off and boxed up to be sent to tions have become more than a Latin Mary Joyce also has other duties but her Mary Joyce at NIS in Riverdale, MD. binomial and specimen in a vial or on first priority is always getting intercep- She checks the Pest ID data on each in- a pin, which makes her job much more tions processed with accuracy and speed terception to make sure it is correct, and satisfying. She has actually become of delivery back to identifiers. There then divides them up according to work quite an insect enthusiast taking a are some older stashes of interceptions location to be returned for identifier’s little extra time to look at the more she has discovered in NIS storage areas collections. This is no small task con- impressive insects that cross her desk. and has been trying to get those finally sidering the mountains of interceptions She sometimes shows NIS staff these back out to the field as well. Summer submitted and processed from all ports interesting specimens and asks questions help comes from student interns, but it by SEL weekly. or reads up about the various kinds of is only for a couple months, so the vast insect pests. She even displays photos of majority of the responsibility falls on Mary Joyce does not see the job is not insects in her cubicle, and has begun to her. We at NIS are very appreciative of just simple mindless data checking take an interest in snails and slugs. Will her hard work, attention to detail, and and paper processing. To her, intercep- nematodes be next? enthusiasm for the job she obviously takes great pleasure in. JF National Identification Services Staff Roles/Responsibilities APHIS Headquarters Riverdale, MD Scott Neitch Tadd Dobbs Administrative and quarantine ac- Identification authority (entomology), Joe Cavey, Assistant Director tion policies, diagnostic reviews entomology identifier specialties Administrative and quarantine ac- tion policies Pete Touhey Mike Petrillo Pest ID technical support, ARM CBP Liaison, cargo release authority Mary Palm, Supervisor technical liaison Administrative, policy, supervision, Joel Floyd special initiatives Steve Bullington Domestic Diagnostics Coordinator, Trend/pathway analysis Joyce Cousins NIS website, NIS INOI editor Urgent process coordinator, pest Indira Singh categorizations, Pest ID technical Botany policy issues, HQ identifier Mary Joyce Burns support orientation and advancement plan, Program Assistant, AQAS quality pest interception training control & interception processing December 20132013 IDENTIFIER IDENTIFIER NOTES NOTES OF INTEREST OF INTEREST NEWSLETTER NEWSLETTER Page 3 Page 3 NIS Urgent Policy Guide Launch

he process by which NIS makes de- The final product of these efforts was Tcisions after urgent pests are identi- sent out to you as a draft for comment fied by national specialists may seem to by Joe Cavey in late October, 2013. It be somewhat of a mystery to identifiers. will be linked from the NIS Identifier For years, NIS staff assigned to urgents Collaboration Intranet site, accessible have used a loose-leaf binder full of only to PPQ. various instructions and policies to assist in making action determinations for While this guide is of most use, of urgents. Historic policies were handed course to NIS in Riverdale, but it down by ancestral staffers like sacred will also come in handy to identi- texts, some of which were lost or needed fiers in making decisions on pests the likes of a Rosetta stone to decipher. for which they have identification The basis for some historical decision authority and during those times memos was not always there or well when NIS is shut down due to articulated. New policies were added to snow storms or other unpredict- address a particular problem that arose, able eventualities. and the binder contents became more complicated to interpret every year. A few caveats to using the guide: 1) start with Chapter 2 for all cargo/conveyance may supersede subsequent charts. It is In an effort to standardize the urgent interceptions; 2) if you have a non- suggested then, that you not go straight process and make it more transpar- reportable organism, you should not be to the flow chart that has the taxa you ent, Bud Petit de Mange, formerly the running it through the flow charts, (but are interested in, at least until you are director of the PPQ Manuals Unit, now there are exceptions, for example, ants more familiar with the overall structure retired, worked with Joyce Cousins and intercepted in Hawaii); 3) the Mexican of the guide. others to document our urgent process Action Policy no longer has species lists, in a flowchart format, similar to other so in most cases, be guided by the quar- Another nice advantage of the guide is PPQ manuals. We had many meetings antine status in Pest ID (and remarks) that the policy documents are linked organized by Joyce to go over the flow when you have a species level identifica- from many of the flow charts, for refer- and try to capture the various aspects tion. You will see there are no longer ence. Just click on the highlighted text, correctly. When NIS staffer, Mike species list for mites, insects, mollusks, and the policy should pop up. Petrillo returned to Riverdale, he started pathogens, or nematodes. (Seeds are working on revising the entire Mexican still listed, i.e., FNWs). The one“list” Take some time to familiarize yourself Action Policy and then facilitated many is for actionable insect genera from with the Urgent Policy Guide since, as more meetings on the overall organiza- Mexico with no known US species; and identifiers, this is a key part of making tion and logic of the way the urgent 4) please realize there is a hierarchy of decisions, in a consistent way, about the manual would flow. these flow charts because one decision important work we do. JF

NIS Staff Roles/Responsibilities (continued) See NIS Staff Directory John McKemy & Megan Romberg Philadelphia, PA National Mycologists identification authority (plant pathology) David Robinson Columbus, OH Steven Passoa National Malacologist, identification National Lepidoptera Specialist Rodney Young & David Bitzel authority (mollusks) National Botanists Beltsville, MD identification authority (botany) Program Manager (vacant) Greg Evans National Sternorrhyncha Specialist Aaron Kennedy Claireann Cook Molecular Biologist, (mycology) Remote Pest Identification Program Page 4 IDENTIFIER NOTES OF INTEREST NEWSLETTER December 2013 The New Nikon DS-Fi2 Camera and L3 Control Unit — First Impressions by Patrick Haslem, Area Identifier, Entomology, Los Indios, Texas e live in a technology-oriented significantly — especially for the less Wsociety where many people go tech-savvy identifiers. The first thing I to great lengths to be the first to try noticed was that the menus have been out any new technological wizardry. redesigned and many of the menu But then there are some people functions familiar from the L2 have (like my parents!) who need a little been relocated in the L3. The best extra time to get used to changes in suggestion I can make for becom- technology, and take longer to learn ing familiar with the new design is even simple tasks using it. I’m about to allot some time for just “playing” in the middle of these two groups with the unit when you first receive — I’m not afraid of new technol- it. Work through the menus one at a ogy, nor am I hesitant to try it, but time until they become familiar. This I usually require a little extra time will take time and patience but is well to feel that I’m using a new product worth the effort. Most of the identi- fluently. I was lucky enough to be fiers who field tested the new units part of the field evaluation team for Figure 1. The camera menu on the monitor of felt very comfortable with the new the DS-L3 can be displayed either on the top the new Nikon DS-Fi2 Camera and design within a couple weeks. right or the top left of the screen. L3 Control Unit that will be provided The Main Menus to the identifiers by the Remote Pest easily within reach, he or she could click The L3 has four main menus accessible Identification Program (RPIP), starting the large camera button with a finger from the image screen CAM (Cam- later this fiscal year. rather than having to use the mouse era), T.BAR (Toolbar), INFO, and required by the L2 . In my case, the SETUP (see Figure 2). The Camera At first glance, the unit is quite similar unit sits further back on the desk than menu includes functions like white to the previous generation Nikon DS- my short arms can reach, so I elected balancing (one of the golden rules of Fi1 Camera and L2 Control Unit. The to keep the mouse (which, by the way, digital imaging), zooming, camera dimensions are almost exactly the same; plugs in at the same exact USB port as gain, scene selection, etc. Clicking the the screen is about the same size as well. on the L2). CAM button once places the menu in Using the same amount of real-estate on the top-right of the screen, clicking it the desktop, the camera will not cause Physical setup is the same with the L3 as again places it in the top left. This is any space deficiencies other than the it was with the L2. No additional cables helpful if you are examining a speci- usual “spaghetti ball” of cables connect- are needed, and when you, the identi- men and need to see the setup menu ing the terminals from the control unit fier receive the camera and control unit but when it is open it obscures part of to the camera, and the computer, plus and place the two side by side, transfer- the specimen. By simply clicking the the power plugs (now if Nikon could ring each cable one at a time from the CAM button, you can then move it to a only invent a wireless electrical plug old to the new units should make setup different location for a more convenient connection . . . !). simple. In fact, if you have the cable view. The T.BAR menu includes func- from the L2 zip-tied or bundled with tions such as distance measuring, color, The most noticeable difference between other cables, it is possible to use all but erasing etc. Clicking the button once the two units will be familiar to those the camera connection cable from the places the menu in the top of the screen, keeping up with the ever-evolving cell L2 with the L3. Another helpful tool clicking it again places the menu in the phone industry — it has a touch screen is the quick setup guide provided by lower 1/3rd of the screen, and click- (see Figure 1). In my opinion, this fea- Nikon (with a few extra tips provided ing the X closes the menu. The INFO ture will be popular among identifiers in a setup guide by RPIP). These give a menu shows the histogram and related who have their cameras/ scopes located pictorial step by step process for setting information about the image as it is on the same desk as their workstations. up the new unit. being viewed on the screen. Finally the If configured properly, the identifier SETUP menu has all of the options for could view the image that is being Okay, now for the meat and potatoes changing the format, size and quality of captured by the camera on one of the of the new unit. This is where the L3 computer monitors, and, if the unit is acquiring the image. and L2 differ, and in some cases, differ continued on next page December 2013 IDENTIFIER NOTES OF INTEREST NEWSLETTER Page 5 used as an independent unit (with images recorded on the CF card, as the current cameras) or can be controlled by Nikon Elements software (with the images captured directly to the com- puter workstation hard drive).

Figure 2. The main menu buttons on the monitor of the DS-L3, including CAM, The L3 is a very sophisticated camera T.BAR, INFO, and SETUP. system that will bring years of imaging The Format Function — It’s worth skimming through Chapter support to the identifier community. Still Important! 8 of the full Nikon Manual that comes With a little experimentation identi- One function that is critical for main- with the control unit, as this chapter fiers can get the settings just right for taining your memory card in good discusses custom settings and what they many of their commonly-encountered working order is FORMAT. Format- do. To manipulate these settings I first specimens. The touch screen func- ting requires a few extra steps with the had to change the camera setting from tion will be a welcome update by new unit (see Figure 3). You must first auto to manual. many, and this interface drives the select the SETUP menu, then click on Nikon system into the current tech- the “Main Menu” button, then click on Another very important feature of the nological times that many of us have the “File” button. There, you will find new system is that the camera can be found a most welcome development. the Format button, on the lower left of the screen.

Adjusting to the New Camera and Control Unit To become proficient using the new camera system it’s helpful to practice us- ing the different settings of the different menus. One way to do this is by taking two sets of images, one using the default settings on the camera, then changing a setting, recapturing, and comparing the images to find out which has a better appearance for what you’re trying to show in your image.

When my camera arrived, the settings were not the same as my L2. I spent some time “tweaking” my settings to get the image size, quality, and format that I wanted. Also, my camera had a tenden- cy to over-emphasize the yellows / reds of my subjects. I found I could correct this by carefully adjusting a few settings within my camera unit. By tweaking the Hue (“purity” of the color), Chroma (color intensity), and Camera Gain red channel (sensitivity of camera to light in the red/yellow range), I did achieve an accurate color rendering. It’s impor- tant that identifiers are aware of what the settings mean so its possible to Figure 3:. Formatting the data card requires a few more steps using adjust them for the best image possible. continued on next page the DS-L3 than the L2. Page 6 IDENTIFIER NOTES OF INTEREST NEWSLETTER December 2013 Using DNA to reveal introduction pathways for fruit flies:I dentifiers’ contribution to comparing interception results from ports of entry with fly captures in the US By Norman Barr and Raul Ruiz-Arce, CPHST Mission Laboratory, Edinburg, Texas In addition to using DNA to identify o protect U.S. agriculture from an interception to species, it is pos- Texotic fruit flies (Tephritidae), sible to determine its likely geographic APHIS PPQ in concert with DHS origin as well. This, however, often Customs and Border Protection insti- requires a thorough understanding of tutes pest exclusion activities such as genetic diversity across a species’ known passenger baggage inspection at ports of distribution and the development of entry. Despite our joint efforts, exotic species-specific protocols. Despite the illustration by Joel Floyd pests are occasionally detected in the investment, these tools significantly U.S. and sometimes result in additional improve our confidence and resolution they share a common pathway. How- surveillance and/or eradication costs of pathways. ever, if the port intercepted material and to the agency. Is it possible to reduce fly captures occurring in the US have The Mission Lab is currently complet- these events? We believe so. One way to distinct genetic profiles, then they may ing the development of methods for enhance our exclusion operations is to have different pathways. pathway analyses of two important identify likely pathways by which flies fruit fly pests: the Mediterranean fruit enter the U.S. and shut them down. In 2012, NIS requested that PPQ en- fly,Ceratitis capitata and oriental fruit This change could be accomplished tomology identifiers submit all suspect flies,Bactrocera spp. For both diagnostic by directing our attention to higher Mediterranean and oriental fruit fly tools, genetic profiles of fly captures risk pathways and targeting increased interceptions to the Mission Lab. These occurring in the US are compared to inspections of problem pathways or specimens include all tephritid larvae potential source populations in Ha- outreach activities along those pathways, that matched the morphological type of waii and other regions. This will help such as traveler education. these two genera. This material is cur- identify likely starting points in the rently being stored in ultra-cold freezers A major obstacle to identifying high pathways. to preserve DNA and its analysis will risk pathways is collecting and analyz- commence in FY14. These interceptions ing the right information. Even though The Mediterranean fruit fly method represent a valuable resource in pathway shipping and host records are important distinguishes among likely pathways of analysis because they provide spatial and for examining trends from interception flies based on worldwide collections. temporal samples for comparison with data, identification of interceptions to This method is currently being tested flies captured over the past few years. species is also needed. Since fruit flies using historical Mediterranean fruit fly Our lab continues to work with identi- are usually intercepted as larvae and not captures from California and Florida. fiers to collect fly material for analyses in as adults at ports, making a definitive The oriental fruit fly method has been future years. Once validated, our goals identification to species level based on used in determining whether Hawaii is include working closely with identifiers morphology is not possible. a possible source of flies captured on the mainland U.S. This project was devel- and state cooperators to ensure that these methods and reference data sets The PPQ Mission Lab in south Texas oped through the cooperation of PPQ provide end users informative diagnostic is continually exploring better DNA S&T, PPQ Hawaii FF Exclusion and tools. methods to help identify pest species Detection Program, USDA-ARS Pacific and support pathway analysis. For Basin Agricultural Research Center, Exposing the true invasion pathway of example, in cooperation with the PIS’ University of Hawaii and the California a pest is a complicated task. It requires in Florida, Texas, and Puerto Rico, the Department of Food and Agriculture. careful consideration of complex Mission Lab conducted a pilot study to This method was successfully applied information and strong collaborations identify intercepted Anastrepha larvae in 2013 to analyze recent California fly among identifiers, scientists, and risk from November 2010 to May 2011. captures. analysts within and outside PPQ. Our That work demonstrated PPQ’s abil- Apart from the host, it is also important ongoing fruit fly projects at the Mission ity to analyze port intercepted material to know the origin of specimens inter- Lab are good examples of how members using standard operating procedures cepted at ports of entry. If these inter- of different PPQ functional areas are and confirmed the identity of important cepted flies share genetic profiles with working together to address program- pest species. the captures in the U.S., then perhaps matic goals.

December 2013 IDENTIFIER NOTES OF INTEREST NEWSLETTER Page 7

Task Force: Molecular Diagnostics at PIS’ (continued from page 1) and NIS policy managers to form the MDTF conducted in-depth analyses of lar biologist with NIS in Beltsville, has MDTF. Their task was to take a broad the logistics, resources needed, policy been working on sequencing various look at the potential for using molecular implications, and other impacts for each intercepted fungi in genera that are diagnostics at port of entry field loca- of the representative test types. These difficult to identify to species using tions to make identifications on cargo analyses were developed to provide PPQ morphology. Other members of this interceptions or to gather pathway leadership the framework that they need group participating at the San Francisco information (see the Dec. 2012 Identi- to determine strategic resource alloca- PIS are Supervisor Art Berlowitz, plant fier Notes of Interest, page 6). tion for using molecular techniques to pathology identifier Fengru Zhang, and detect and identify pests at ports. technicians Shana Gallant and Kyle First, the three working groups exam- Beucke. This pilot will sequence inter- ined in detail 1) the breadth of pest or- A template was developed and used ceptions of Colletotrichum from San ganisms which this technology could be for each analysis in order to cover all Francisco and other California ports applied to; 2) possible criteria for devel- the aspects, impacts, and implications of entry to ascertain what species are oping and using molecular diagnostics of the representative test type being coming in on various hosts. C. gloeospo- for Agriculture Quarantine Inspection analyzed. For each pilot, each detailed rioides is currently considered non-re- (AQI) purposes; and 3) currently avail- implementation plan included: hosts, portable morphologically, but is actually able molecular assays including finding specificity, logistics, policy implica- a complex of twenty-two species when out the extent to which other country’s tions, costs of equipment, supplies, looked at with DNA analysis. Some national plant protection organizations physical space needed, training and sequencing data for Colletotrichum use molecular diagnosis for making personnel requirements, interactions have already been analyzed from port regulatory decisions. with CBP, trade implications, benefit to interceptions and determined at NIS PPQ, needed collaborations, and other Beltsville, but this pilot seeks to gather Preliminary discussions were held with considerations. The very detailed plans more data and answer the question if the PPQ Management Team, and an were presented to the PPQ Manage- some of these species should actually be operational plan was developed at their ment Team in August, and after a lively considered reportable/actionable. request. Fulfilling aspects of the Opera- discussion, they approved each of the tional Plan is the effort that the group pilots for immediate implementation Thrips Sequencing Pilot: The thrips pi- has been working on since the Spring .We are moving forward in purchasing lot will take place at the Miami PIS with of 2013. The highest priority pests were supplies and planning for training at the Cheryle O’Donnell, now an identifier then determined and grouped according various work locations. in San Diego, as the pilot lead work- to the type of tests. The MDTF identi- ing with CPHST Mission molecular fied representative test types, each of The three pilots were chosen because biologist, Norman Barr. The pilot builds which could be used for a range of pests. they represent a variety of molecular on work already begun by Identifier The test types chosen represent different diagnostics technologies and each are Tom Skarlinksi, in cooperation with a aspects of what molecular diagnostic real-world questions or needs for PPQ researcher at the University of California techniques can do to assist PPQ in the interception diagnostics and identi- at Riverside, has been looking at identi- goal of accurate, timely, and reliable fication. The main objectives revolve fications of immature thrips, primarily identifications as well as pathway data around the feasibility of doing this kind Franklinella spp., frequently intercepted gathering. of work at PIS’, and the potential infor- on cut flowers from South America. mation gained from each of these pilots Tom found some morphological differ- After much telephone collaboration of represent secondary objectives. The ences in larve based on re-examination the working groups, the test types finally pilots as planned are the following (click after molecular identification. The chosen for pilot studies are Polymerase on the pilot title for more details): molecular identifications will help PPQ Chain Reaction (PCR) and DNA determine if immature thrips are action- sequence analysis for fungi (using Col- Colletotrichum Sequencing Pilot: This able or not, thus potentially reducing letotrichum), for insects (using thrips), pilot project will take place at the San the numbers of fumigations required. and detection assays such as immune- Francisco PIS, and focuses on DNA test strips and a new technology called sequencing of interceptions of fungal Sequence data from interceptions and CANARY (using Ralstonia solanacear- pathogens in the genus Colletotrichum. laboratory methods will be developed um, a bacterial plant pathogen). The The pilot lead, Aaron Kennedy, molecu- continued on page 8 Page 8 IDENTIFIER NOTES OF INTEREST NEWSLETTER December 2013

Molecular Diagnostics Task Force IDP Facilitation The PPQ identifier position was -de (continued from page 7) (continued from page 1) veloped and first staffed more than 45 at UC Riverside and shared with the years ago, specifically to facilitate the 2. Revising PPQ policies, pro- Miami PIS over time, to eventually be clearance of imported cargo at a time cedures and staffing to facilitate ID able to implement the procedures on when overnight mail service did not ex- processing. their own. Besides Tom, other mem- ist. Since then, a small identifier cadre bers of the Miami PIS team involved has grown, with expanding international Delays may result from local circum- include PIS Supervisor Pedro Milan, trade and domestic programs, and will stances, such as the absence of a PPQ the new Plant Pathology Identifier, soon exceed 100 in number. You, the identifier or lack of sufficient numbers Weston Msikita, and biological techni- current identifier cadre, efficiently of inspectors at the site. Overarching cians. Ultimately, the information clear 85-95% of cargo shipments held issues may cause delays at multiple port gathered from immature thrips se- for intercepted pests through com- of entry locations. For example, delays quencing will help inform the desig- mendable and consistent performance. in identifications of Prompt and Rou- nation of consistent morphological But, as cargo volume and clearance tine interceptions made at the national characters that can be used for species responsibilities continue to increase, level can slow issuance of ID Authority level identifications. this task becomes more difficult. The which, in turn, affects cargo clearance IDF project and associated hiring plan time adversely. NIS and FO manag- Ralstonia solanacearum CANARY will be implemented to help PPQ meet ers are working with CBP and you to Pilot: The two PIS’ were chosen for current and future timeliness require- identify and resolve these issues. this pilot are Atlanta, GA and Linden, ments for pest interception processing NJ. because of their volume of gera- while maintaining the credibility of pest The IDF plan includes a number of nium, or Pelargonium, plant imports identifications. internal policy and procedural changes from countries not part of the off-shore that can help facilitate time for identi- certification program. This host show fication without sacrificing credibility. no symptoms while harboring the bac- While PPQ may find that we do not Welcome Recently Hired New terial pathogen, Ralstonia solanacearum, need to implement all of the changes race 3, biovar 2 which can also infect Identifiers listed in the IDF plan, some were iden- potato, tomato, and eggplant among tified as critical some time ago. Revi- others. It is a serious quarantine patho- Plant Pathology sion of our ID Authority policies consti- gen also on the Select Agent list. tutes one of these areas. In January, NIS Portland. OR Jinya “Jack” Qiu plans to implement revisions of IDA, The pilot lead is Laurene Levy, who Miami, FL Weston Msikita especially for non-reportable groups and with CPHST Beltsville molecular for Provisional IDA. Changes will ex- biologists, Zhaowei Liu, Gang Wei, Entomology pand conferred entomology and botany and Jinbo Wang have been working on IDA, primarily. Another potential the CANARY technology for a number Miami, FL Alexander Cunningham objective involves modifying submission of years. The test is a serological assay Buffalo, NY Carla Coots procedures for botany interceptions to designed to detect R. solanacearum (but increase use of digital imaging. PPQ not the race/biovar) from cuttings im- may also look into how we might maxi- ported through the PIS’. The PIS par- Recent Newly Relocated Plant mize the contributions of technicians ticipants include Atlanta Plant Health involved with processing interceptions Inspection Stations: Safeguarding Officer, Cesar Cardona, and the use of digital imaging, in gen- Linden PIS Supervisor Randy Cadet eral. And finally, discussions with you Atlanta, GA and others. The purpose of the pilot is are planned for the IDF project. NIS Los Angeles, CA to see if this technology can be imple- and FO will contact identifiers at most Nogales, AZ mented effectively at a PIS, if it can ports to discuss local pest interception detect R. solanacearum in plant imports processing efficiency. We want to hear and blind tested positive controls, and your assessment and ideas for improving how it compares with immunostrip the process at your location and within technology, currently used on some your area of coverage. imported Pelargonium plants. December 2013 IDENTIFIER NOTES OF INTEREST NEWSLETTER Page 9 Entomology Specialists Star in Instructional Videos for Identifiers ideo products of a 2012 Farm Bill by Joel Floyd Vsuggestion are starting to appear on a computer monitor near you. Due to the difficulties with getting travel funds and organizing pest identification workshops, an alternative approach is being tried by video taping training and posting them on the web. The first of these are8 videos (click here) on techniques for mounting soft-bodied in- sects on microscope slides, including one on mailing them. These were initiated by Gary Miller, SEL Aphididae Specialist, and now Research Leader at the USDA-ARS Systematic Entomology ARS-SEL Research Leader Gary Miller in one of the Laboratory (SEL). Also included is a video on mounting mites, instructional videos on how to mount insects on slides. with Ron Ochoa. They are now posted on our NIS Identifier Col- laboration Intranet site. The videographer and editor is Arthur Tracy, who coincidentally is the son of Area Identifier Bob Tracy at Linden, New Jersey. The second group of videos are still in the editing process at the University of Florida and include lectures, student questions, and keying out exercises. The first group includes Aleyrodidae, Aphididae, and Coccoidea conducted by John Dooley and Greg Evans (PPQ), Gary Miller (SEL), Susan Halbert and Ian Stocks (Florida Division of Plant Industry). The second group included Auchenorrhyncha (Fulgoroidea, Membracoidea, Cercopoidea), and Pentaomoidea taught by SEL’s Stuart McKamey, Charles Bar- tlett of the University of Delaware, and Joe Eger of Dow Agrosci- ences. We only had small groups of students who included a few area identifiers who have these taxa as specialties. Thanks much goes to Amanda Hodges, Director, Doctor of PPQ Specialist Greg Evans showing insect morphology Plant Medicine program at University of Florida, Gainseville for during filming of a taxonomic instrutional video in organizing this major effort, with help from Stephen McLean and Gainsville, FL. Photo courtesy of Julieta Brambila Stephanie Stocks. These videos should be ready in early 2014.

Entomologist Joe Eger during filming of video on Screen shot of SEL Specialist Stuart McKamey from edited pre- Pentatoidea keying out instructions. sentation on Auchenorrhyncha taxonomy. Photo courtesy of Stephanie Stocks Page 10 IDENTIFIER NOTES OF INTEREST NEWSLETTER December 2013 by Deena Walters, PPQ Identification Technology Program (ITP) imageID Cooperator at Colorado State University, Ft. Collins, Colorado bout a year ago, the Identification the pest(s) in an image has been verified those accurately-identified images that ATechnology Program (ITP) in Fort by an expert. At Encyclopedia of Life represent the taxa in the PestID system. Collins, Colorado received Farm Bill (EOL), they use the phrase “trusted” Instead of identifiers having to this on funding to begin work on an image to indicate that an image has been so their own, we are creating a searchable database and website based on pest im- verified. website containing such images. ages taken over the years by Identifiers. We call the project imageID. It is being A large group of images that automati- Revealing relationships developed in cooperation with Dr. Mark cally have been verified are those of type During our interactions with identifiers Simmons of Colorado State University specimens. A type specimen is the over the past year, sometimes in person and Joe LaForest of the University of specimen that a taxon name was origi- and also by way of e-mail and question- Georgia. nally based on. So, the identification of naires, we’ve been given a clearer picture images of these specimens are inherently as to the functionalities that identifiers The purpose of the website imageID correct, although sometimes labelled would like to see in imageID. Since is to give all identifiers access to each with an outdated synonym given the some pests are relatively specific when other’s images, but particularly those ever-changing nature of plant and it comes to association with a particu- images of diagnostic value. Diag- lar crop import, especially when nostic images are those that aid in originating from a particular part the identification process because of the globe, we have begun an they show relevant characters and analysis of the years of data in character states. Typically, no one the PestID system as it relates to image is going to allow someone region/commodity/pest associa- to identify an unknown pest all tions. Results of these analyses the way down to species. So, will be incorporated into the what we are doing is collecting website in a way that is useful for and displaying suites of images identifiers. For example, a search that, when taken together, point on the pest group “Coleoptera,” an identifier in the right direction the commodity “mango,” and the or, at the very least, narrow down region of origin as “Central Amer- the possible taxon identifications. ican & the Caribbean” would bring up a note saying that “. . . What’s trustworthy? 95% of past Coleoptera intercep- Although imageID began as a tions have been of the pest species way to let identifiers share their Stenochetus mangiferae.” images, plans for the project have since evolved into what we see Size Matters as an identification aid offering Other concerns expressed by more than just the identifiers’ identifiers that we are address- images. As many of you know, ing include: 1) the inclusion of there are innumerable images synonyms and their direct links in on the internet of various plant pests. animal nomenclature. In any case, vari- the database; 2) coding of images based Unfortunately, there is also considerable ous organizations, including the MCZ on life stage and body parts so that an variation in the quality of those images Type Database @ Harvard Entomology, identifier can focus on a narrowed group for identification purposes. For example, are offering digital images of the type of images (this is called “filtering”); 3) images need to be clear, sufficiently large specimens in their collections. including labelled organism outlines to see desired details, and perhaps most for various pest groups since body part importantly, they need to be correctly My point is that there are many im- names can vary across these groups; and identified. The latest trend at some of ages on the internet that are of value 4) having a few organism-specific char- the websites featuring pest images is to for pest identification. The trick is to acters than can be used for filtering im- say whether or not the identification of weed through all the junk and find ages. With respect to the latter, I’ve been December 2013 IDENTIFIER NOTES OF INTEREST NEWSLETTER Page 11

working with Charles Brodel on some Jarmin, Liana Maller, and Rodney and fruits last spring. We’ve taken de- basic distinguishing Coleoptera charac- Young. The plan in the future will be to tailed images of this material, focusing ters that could reduce a larger selection put out a “call” for images of particular on characters important for identifica- of images to those more likely to be of pest taxa, instead of trying to choose tion, for which we often provide label- use to an identifier trying to identify a images from a particular person’s col- ling on the images as well as detailed particular unknown. It can be as simple lection. Late this summer, we put out a image descriptions. as selecting a size category, e.g., beetles request for images of various Coleoptera < 3 mm long VS. beetles 3-13 mm long taxa, along with instructions on how to What’s ahead? VS. beetles 13-20 mm long VS. beetles transfer these images to our FTP site. Currently, we are developing the image- > 20 mm long. We also plan to offer at ID website from several angles, all em- the website pertinent screening aids that In addition to using images created by phasizing the usefulness of images to the have been produced by various experts the identifiers, various other scientific pest identification process. This is the over the years. experts, and reliable sources on the end of year one of what was proposed as internet, we have also begun a program a three-year project. However, we hope The first identifier collection of images of taking high-quality images of inter- to have a beta-version online for testing I evaluated belonged to retired identi- cepted pests that have survived well in in June of 2014. We will let you know fier Bill Carlson. More recently, we’ve storage over the years. These pests are when the website becomes available. In received images from several identifiers, mostly plant propagules such as seeds or the meantime, any suggestions anyone including Greg Bartman, Patrick Mar- fruits. Our trial program with Shannon may have are certainly welcome and can quez, Alan Smith-Pardo, Phil Johnson, Jarmin at SFO has gone very well. She be sent to me at : John Dooley, David McCoy, Shannon sent us close to 100 intercepted seeds [email protected]. Page 12 IDENTIFIER NOTES OF INTEREST NEWSLETTER December 2013 Identifying Beetle Larvae from Wood Packing Material Hannah Nadel, CPHST Otis Laboratory, Buzzards Bay, MA and Peter Reagel, Xavier University, Cincinnati, OH fter establishment of Asian long- Ahorned beetle and emerald ash borer in North America, International Standards for Phytosanitary Measures #15 (ISPM-15) for solid wood packing materials (WPM) were adopted in 2005 to reduce risk of further introductions of tree and timber pests. WPM is treat- ed before export with heat or methyl bromide fumigation at certified facili- ties and marked with an IPPC stamp to signal compliance. But live wood borers are still routinely intercepted in marked WPM at US ports, indicating ineffective phytosanitary treatment or counterfeit marking.

Because most wood boring insects are intercepted as larvae, which are at best difficult to identify while vessels and Figure 1. CBP agriculture specialists examine wood packing material cargo are awaiting clearance, they gener- at Miami seaport. ( All photos courtesy of Hannah Nadel) ally trigger re-export based on family level identification. However, valuable data can be used to further streamline quences from identified adults are regulatory and pathway risk information targeted inspection of WPM from treat- submitted to database libraries GenBank is lost when larvae remain unidentified. ment facilities consistently exporting and BOLD, they can generally be used Documentation of consistent treatment higher-risk species, in addition to shed- to identify conspecific or congeneric failure for identified high risk species ding light on the validity of treatment barcodes submitted earlier or later in can be used to direct phytosanitary marks. Currently, unidentified larvae time, and from any life stage. research toward improving treatment may be sent to the USDA-ARS Sys- efficacy for those species, and pathway tematic Entomology Laboratory (SEL), Project scope was limited to the Ceram- where systematic knowledge for bycidae and Buprestidae because Otis identification below the fam- Laboratory has the potential to rear ily or subfamily level is usually members of these families on artificial limited or unknown. diet, if necessary. The laboratory is in a unique position to carry out this project In 2012, with funding from AQI not only because of its rearing capacity and cooperation with Assistant under PPQ permit and containment, Professor Ann Ray at Xavier but because it houses a DNA laboratory University, Cincinnati, OH, the and expertise in phytosanitary wood CPHST Otis Laboratory in Buz- treatment. After a six-month pilot zards Bay, MA, took up the chal- study in Seattle coordinated with ento- lenge to improve identification mology identifier Emilie Bess and CBP capacity for intercepted larval agriculture supervisor Alisha Beckham, wood borers. Two complemen- the handling, shipping, and rearing pro- tary approaches are used: larval Figure 2. At the CPHST Otis Laboratory tocols were developed and the project rearing to the identifiable adult was expanded to include Long Beach, under containment, infested host wood stage and expansion of DNA CA, Detroit/Romulus, MI, and Hous- is held in metal mesh bags and checked barcode databases from identi- ton, Laredo, and Pharr/Hidalgo, TX. fied adults. Once barcode se- periodically for adult emergence. continued on next page December 2013 IDENTIFIER NOTES OF INTEREST NEWSLETTER Page 13

Figure 3. A cerambycid larva that left its host wood Figure 4. Larvae are reared singly in cups of artificial and is being reared on artificial diet. This larva was diet under controlled conditions with containment. intercepted at Long Beach with tile cargo from Italy.

In October 2013, the ports of Miami visible); the rest died in transit, dur- treatment types. For example, the Asian and Port Everglades, FL, also signed on. ing rearing, or were hidden in wood. longhorned beetles arrived on separate The work is carried out by a large team. Rearing is still in progress for about 80 vessels but in WPM bearing the IPPC CBP agriculture specialists (Fig. 1) bring larvae. Of 160 dead cerambycid larvae, mark of a single wood treatment facility live cerambycid and buprestid larvae 75% of them were identified to 16 spe- in China and indicating treatment with and ideally up to 6 inches of host wood cies and 11 genera, while the remainder methyl bromide. to the PPQ identifiers, along with the matched no barcode even at the genus completed PPQ form 309 and IPPC level in the databases. About 70 ceram- The next phase of the project will also stamp information. Action status at the bycid adults were reared, of which more include wood identification at Penn- port is carried out as usual. A kit sup- than 30 examined so far at SEL were sylvania State University to further plied by Otis Laboratory is used to ship identified to 14 species in 10 genera. examine risk factors. The project is the insects overnight to the Otis Insect Combined larval and adult identifica- expected to continue on its present Containment Facility where they are tions total 19 species and 14 genera. course through 2014 and will likely be reared, preferably in the tunneled host extended beyond, with future tabling of wood (Fig.2), but in artificial diet if host Notable cerambycid interceptions options to shift the focus to particular wood is not supplied (Figs. 3,4). Reared include known pests such as Asian commodities and other pest taxa. adult tissue is retained for DNA barcod- longhorned beetle, Anoplophora glabri- ing at Otis Laboratory, and pinned pennis (5 interceptions), and velvet specimens (Fig. 5) are sent to SEL longhorned beetle, Trichoferus for identification or confirmation by campestris (6 interceptions), morphology. Tissue from larvae that die although several other species during transit or rearing is also retained have pest potential. To date, at Otis Laboratory for DNA barcoding 35 buprestids were received and the sequences are compared with but none was reared to adult, others in databases for potential conspe- and DNA barcoding has just cific or congeneric matches. begun. Overall, early project results are promising. Risk To date about 345 intercepted ceram- pathways are beginning to bycid and buprestid larvae from 19 emerge that should result countries were received by Otis Labora- in targeted inspection of Figure 5. Asian longhorn beetle reared from a tory; of these, over 140 cerambycid and wood coming from particu- larva intercepted in Seattle (note that two legs 13 buprestid larvae appeared healthy (if lar treatment facilities and were removed for DNA analysis). Page 14 IDENTIFIER NOTES OF INTEREST NEWSLETTER December 2013

Plant Quarantine Inspectors, Port of Philadelphia, 1926 Photo courtesy of the APHIS Library

Entomological Workers at the US National Museum, Washington DC, 1925 Photo courtesy of the Smithsonian Instituion Archives