PROCEEDINGS OF THE

2017 Annual Imported Fire Ant and

Other Invasive Ants Conference Mobile, AL

May 16-18, 2017

Hosted by:

Auburn University College of Agriculture

and

Alabama Cooperative Extension System

Planning Committee:

Fudd Graham

Kathy Flanders

Anne-Marie Callcott

Tim Davis

Vicky Bertagnolli

Kelly Palmer

Local Arrangements, Registration and Setup

Henry Dorough

Lucy Edwards

Ellen Huckabay

Mallory Kelley

Jeremy Pickens

Special Thanks to Renaissance Mobile Riverview Plaza Hotel

Kelly McDonald

Cadi Mills

Jenney Granata

Chiquitta Howard

Nora Presswood

Special Thanks to Admin Staff at Auburn

Will Rankin

Thank you Will for managing funds for the meeting, paying bills, ensuring we

stayed within AU guidelines and putting up with Fudd

These proceedings were compiled from author submissions of their presentations at the 2017 Imported Fire Ant

and Invasive Pest Ant Conference In Mobile, AL. The opinions, conclusions, and recommendations are those of

the participants and are advisory only. Mention of trade names or commercial products in this publication is sole-

ly for the purpose of providing specific information and does not imply recommendation or endorsement. The

papers and abstracts published herein have been included as submitted and have not been peer reviewed. They

have been collated and duplicated solely for the purpose to promote information exchange and may contain pre-

liminary data not fully analyzed. For this reason, the authors should be consulted before referencing any of the

information printed herein. This proceedings issue does not constitute a formal peer review publication.

Many thanks to our sponsors

Platinum Sponsors

Silver Sponsors

Table of Contents Past, Present and Future: written submissions were not requested from invited speakers History of the Annual IFA Research Conference and thoughts for the future. Tim Davis (University of Georgia Extension)………………………………………………..1 Historical perspective on Clemson’s contribution to the fire ant research and Extension. Paul M. (Mac) Horton (retired Director Clemson Sandhill Research and Education Center) – invited……………………………………………………………..NS The 3 years of fire ant chemical ecology that turned into 40, and still counting. Robert Vander Meer (USDA-ARS-CMAVE) – invited……………………………..........NS Imported Fire Ants: What they do and why they do it. Sanford Porter (USDA-ARS-CMAVE) – invited…………………………………………………………...NS History of APHIS IFA quarantine. Anne-Marie Callcott (USDA-APHIS-PPQ)……………8 General Presentations: Biosecurity and surveillance of ants in Australia. Antonette Walford (Department of Agriculture and Water Resources. Australia)………………………………………..........15 Red imported fire ants in Australia: Update. Robert T. Puckett (Texas A&M University, Dept. of Entomology, Texas A&M AgriLife Extension)…..…………………..NS Systems approach to imported fire ant compliance inspection. Awinash Bhatkar (Texas Dept. of Ag)………………………………………………………………………….NS Watching ants: How insect behavior impacts protocol. Gabriela Perezchica-Harvey and Jennifer A. Henke (Coachella Valley Mosquito and Vector Control District)………….NS Imported fire ant worker size as relates to species/hybrid and geographic distribution in Tennessee. Manoj Pandey (Tennessee State University), Jason Oliver (TSU), Karla Addesso (TSU), Steven Valles (USDA-ARS-CMAVE), Reginald Archer (TSU), and Anne-Marie Callcott (USDA-APHIS-PPQ)……………………………………..16 Volatile compounds in the imported fire ants. Jian Chen (USDA-ARS-NBCL)…………...NS Fifteen years with the Lago Santa Fe fire ant project. Paul Nester (Texas A&M AgriLife Extension Service)………………………………………………………………….18 Evaluation and Review of the Scotts® Electric Powered Handseeder – the Wizz™. Timothy S. Davis, PhD. (University of Georgia Extension Agent – Chatham County)…….25

Geographic incidence of Kneallhazia solenopsae and Solenopsis invicta viruses (SINV-1, -2, and -3) in imported fire ant species and their hybrid in Tennessee. Jason Oliver (Tennessee State University), Karla Addesso (TSU), Steven Valles (USDA-ARS-CMAVE), Nadeer Youssef (TSU), Manoj Pandey (TSU), Paul O’Neal (TSU), Joshua Basham (TSU), Joseph Lampley (TSU), Debbie Eskandarnia (TSU), Garrett Roper (TSU), Megan Patton (TSU), Steve Powell (Tennessee Dept of Agriculture), and Anne-Marie Callcott (USDA-APHIS-PPQ)………………………………27 Introduction and establishment of fire ant biocontrol agents in the Coachella Valley of California. David Oi (USDA-ARS-CMAVE), Steven Valles (USDA-ARS- CMAVE), Sanford Porter (USDA-ARS-CMAVE), Christopher Cavanaugh (Coachella Valley Mosquito & Vector Control District) and Gregory White (Coachella Valley Mosquito & Vector Control District)……………………………………30 Tropical fire ants and Pseudacteon decapitating flies in Ecuador. Sanford Porter (USDA-ARS-CMAVE), Henri Herrara (Escuela Superior Polite̒cnica de Chimborazo, Ecuador), Robert Vander Meer (USDA-ARS-CMAVE), and Charlotte Causton (Charles Darwin Foundation for the Galapagos Islands, Ecuador)…………………………NS The USDA-APHIS phorid fly release project: 2001-2017. Anne-Marie Callcott (USDA-APHIS-PPQ) ……………………………………………………………………….33 Getting familiar with invasive ants. Margaret L. Allen (USDA-ARS)…………………....NS The tawny crazy ant (Nylanderia fulva) at the port of Savannah, Georgia. Ben Gochnour and Dan Suiter (University of Georgia)…………………………………………37 Alleged injury to new born calves in Texas counties attributed to presence of tawny crazy ant. Paul Nester and Robert Puckett (Texas A&M AgriLife Extension Service)……38 Fire ant data management: Preserving the past and tools for the future. Joseph LaForest (University of Georgia), Rebekah Wallace (University of Georgia), and Kathy Flanders (Auburn University)…………………………………………………...NS Past, present and future of eXtension IFA/Ant Pests communities of practice. Kathy Flanders (Auburn University) ……………………………………………………...NS Posters: Fire ant control demonstration at Storybook Farm. Mallory Kelley (Alabama Cooperative Extension System) and Fudd Graham (Auburn University)……………………42 Evaluation of various insecticides and insecticide combinations as fire ant quarantine treatments on commercial grass sod. Kelly Loftin and John Hopkins (University of Arkansas Extension System), and Anne-Marie Callcott (USDA-APHIS-PPQ)……………..45

Contact and fumigation toxicities of three isothiocyanate compounds from Bagrada Bugs against red imported fire ants and tawny crazy ants. Yuzhe Du, Jian Chen and Michael J. Grodowitz (USDA-ARS-NBCL)………………………………..NS Evaluation of bifenthrin for control of tawny crazy ants (Nylanderia fulva) in container substrates. Jeremy Pickens (Department of Horticulture, Auburn University), L.C. (Fudd) Graham (Department of Entomology and Plant Pathology, Auburn University), and Kelly Palmer (Alabama Cooperative Extension System)………….……..NS An estimation of temperature limits for the tawny crazy ant, Nylanderia fulva (Hymenoptera: Formicidae). David C. Cross and M.A. Caprio (Mississippi State University)…………………………………………………………………………………..49 Comparative water relations and desiccation tolerance of four invasive ant species. Olufemi Ajayi (Auburn University), Arthur Appel (AU), Li Chen (Chinese Academy of Sciences), Rammohan Balusu (AU), and Henry Fadamiro (AU)………………………..56 Relationship of imported fire ant (Solenopsis spp.) integument coloration to cuticular hydrocarbon and venom alkaloid indices. Manoj Pandey (Tennessee State University), Jason Oliver (TSU), Karla Addesso (TSU), Steven Valles (USDA-ARS-CMAVE), and Anne-Marie Callcott (USDA-APHIS-PPQ)…………………57 List of Attendees…………………………………………………………………………….58

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History of the Annual IFA Research Conference and Thoughts for the Future

Facilitated by Dr. Tim Davis University of Georgia Extension Service

The Annual Imported Fire Ant and Invasive Ant Conference has been slowly changing in the last 5-10 years. Attendance has dropped and many of us involved in IFA work of all aspects, have found our IFA work to be lessened as we are assigned other duties. Resources, both financial and staffing, are decreasing in the IFA community. Therefore, instead of one key-note speaker this year, we decided to invite several of those in our group who have been with IFA for multiple decades to give us a perspective of their work. The planning committee would especially like to thank the following for coming and giving us an overview of their 30-40 years of work with IFA.

• Dr. Paul M (Mac) Horton, retired, Clemson University • Dr. Robert Vander Meer, USDA-ARS • Dr. Sanford Porter, USDA-ARS • Anne-Marie Callcott, USDA-APHIS

In addition, we, as a group, spent a good bit of the conference talking about and deciding how to move forward into the future as a group. We thank Tim Davis for leading these conversations. As a result, the group has decided:

• Change the name of the conference to Invasive and Pest Ant Conference (IPAC) • Meet with the NCUE-National Conference on Urban Entomology– during their meeting

years (every other year, 2018, etc). David Oi will facilitate this for 2018 • On the off year of NCUE, instead of a formal conference, perhaps have field working

days or other similar educational events On the following pages is a presentation by C.S. Lofgren, made 30 years ago, for the 1987 Imported Fire Ant Conference. This text provided a brief overview of the previous 25 years of IFA conferences (prior to 1987) and had a listing of the conferences from 1963-1987. We have added to that table to show the meetings through 2017.

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A Quarter Century of Imported Fire Ant Research Conferences From Proceedings of the 1987 Imported Fire Ant Conference

C. S. Lofgren USDA-ARS

At the March 1986 Imported Fire Ant Conference, I was asked to prepare an historical review of the origin and development of these conferences. With the help of Mr. W. A. Banks I was able to determine the location, dates, and chairman of the meetings since their origin in 1963 and various other miscellaneous information. Before elaborating on this information, however, it is important to recognize that research on imported fire ants (IFA) pre-dates these conferences. Of particular interest is the work of Wilson and Eads (1949). Their report presents the first biological observations on the IFA in the U.S. The USDA established a research station in Spring Hill, Alabama in 1949. Surveys by Mr. George Culpepper while at this laboratory from 1949-1953 revealed for the first time the extent of the IFA infestation and its link with nursery stock. Control techniques using chlorinated hydrocarbons were developed at Auburn and Mississippi state universities and at the USDA lab. A more detailed account of this early history is published in the proceedings of the Fire Ants and Leaf-Cutting Ant Conference held in Gainesville, FL in March 1985 (Lofgren, C. S. 1986. History of the Imported Fire Ant in the United States. pp. 35047. In Fire Ants and Leaf Cutting Ants: Biology and Management. C. S. Lofgren and R. K. Vander Meer (eds.). Westview Press, Boulder, co, 435 pp.). The first large scale effort to control IFA started with the Federal-State Control Program initiated by the U.S. Congress in 1957. At that time I became involved in IFA research when I was transferred from the ARS Insects Affecting Man and Animals Research Laboratory in Orlando, FL to the Plant Pest Control Division (PPCD) of ARS in Gulfport, MS to organize an IFA Methods Development Laboratory for the purpose of developing improved methods to control IFA. After several years it became evident that we needed more direct exchange of information between this laboratory and other IFA researchers. Communications with Dr. Murray S. Blum, who was at LSU at that time, and Mr. Henry Green, Mississippi State University, resulted in the decision to hold a workshop in Gulfport in 1963. In July of that year I transferred back to the Insects Affecting Man and Animals Research Laboratory (now located in Gainesville, FL); however, other entomologists at the IFA laboratory, Mr. W. A. Banks, Mr. F. J. Bartlett, and Mr. C. E. Stringer, continued contacts with Dr. Blum and Mr. Green and the first conference was held in Gulfport, MS in October, 1963. Correspondence between Mr. C. C. Fancher, Regional Supervisor of PPCD, ARS, and Dr. Carroll N. Smith, Laboratory Director, Insects Affecting Man and Animals Research Laboratory, Outline the agenda and organization. The letters are reproduced on the next 2 pages.

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At the termination of the first meeting, Dr. M. S. Blum was elected chairman for the next meeting in 1964. Unexplained delays resulted in this conference being held in February, 1965 in Gulfport, Mississippi. The third conference was hosted by Mr. H. B. Green at Mississippi State University in November, 1965. There is no record of a meeting in 1966, but a conference was held in Biloxi, MS in March 1967. Correspondence in our files indicated that this meeting was to be hosted by Mr. W. C. Rhoades, University of Florida, at Quincy, FL; however, travel restrictions caused it to be moved to Biloxi. A meeting announcement from Mr. Carl Scott, Georgia Department of Agriculture, stated that the next conference, which was held in Biloxi in March 1968, was the 5th Annual Research Conference. From this time forward a conference was held in all years except 1985. In 1985 the conference was to be held in Puerto Rico; however, the chairman appointed did not fulfill this commitment. A late attempt was made by Dr. S. B. Hays, Clemson University, to organize a conference, but lack of commitments by persons to attend caused its cancellation. A list of all the conferences, locations, chairmen and number of attendees is given in Table 1*. The initial guidelines for the conferences specified that no proceedings would be published. These guidelines were changed in 1984. At this time, a committee chaired by Dr. Mike Mispagel assumed responsibility for assembling a non-refereed proceedings of abstracts, short papers, etc. Prior to this time unofficial notes, papers, and abstracts were found in our files for the years 1967, 1973, and 1980. A review of the information in Table 1 reveals two periods when attendance at the meeting increased significantly: 1972 to 1975 and after 1978. The first of these coincides with a period during which grant money was provided by the USDA to university personnel. The second occurred following the cancellation of registrations for Mirex by the EPA. The latter resulted in a large number of attendees from commercial concerns interested in IFA control. In conclusion, I have kept all the records, sparse as they are, at this office. If anyone reading this report has any information on the conferences, I would appreciate receiving a copy for the files. *The compilers have taken the liberty of reformatting Table 1 from Lofgren, eliminating the chairmen, and adding the dates from 1987-2017 to the table. To see the original, please go to the 1987 Proceedings available online at: http://articles.extension.org/pages/19000/proceedings-of-the-imported-fire-ant-conference

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Table 1. Information on IFA Conferences from 1963-2017. Re-formatted from Lofgren 1987 IFA Proceedings and information for 1987-2017 added by 2017 conference compilers.

YEAR DATES LOCATION # attendees # talks # posters 1963 Oct Gulfport, MS 15-20

1965 Feb Gulfport, MS --

1965 Nov Starkville, MS --

1967 Mar Biloxi, MS 27

1968 Mar Biloxi, MS 25

1969 Mar New Orleans, LA 30

1970 Apr Gainesville, FL --

1971 Mar Gulfport, MS 33

1972 Mar Athens, GA 62

1973 Mar New Orleans, LA --

1974 Mar Brownsville, TX 38

1975 Mar Gainesville, FL 50

1976 Mar Gulfport, MS 64

1977 Mar College Station, TX 54

1978 Apr Raleigh, NC 83

1979 May Baton Rouge, LA 89

1980 Mar Gainesville, FL 78

1981 Mar Biloxi, MS 85

1982 Mar Austin, TX 97

1983 Mar Starkville, MS 122

1984 Mar 27-28 Gainesville, FL 126 32 0 1986 April 3-4 Austin, TX 97 25 0 1987 April 15-16 Baton Rouge, LA 63 22 0 1988 May 4-5 Athens, GA 63 24 0 1989 April 18-19 Biloxi, MS 95 24 0 1990 April 2-3 College Station, TX 115 29 4 1991 March 19-20 Atlanta, GA 83 15 0 1992 April 20-24 San Juan, PR 41 20 0 1993 June 15-18 Charleston, SC 84 31 0 1994 May 9-11 Mobile, AL 99 24 0 1995 May 2-4 San Antonio, TX 111 38 0 1996 April 16-18 New Orleans, LA 111 33 2 1997 April 14-16 Gainesville, FL 89 25 9 1998 April 6-8 Hot Springs, AR 176 45 0 1999 March 3-5 Charleston, SC 169 30 17 2000 April 5-7 Chattanooga, TN 160 50 25 2001 Feb 28-Mar2 San Antonio, TX 192 49(16+33)* 34 2002 Mar 24-26 Athens, GA 168 34 33

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YEAR DATES LOCATION # attendees # talks # posters 2003 Mar30-April 1 Palm Springs, CA 155 29 36 2004 Mar 21-23 Baton Rouge, LA 137 32 22 2005 Mar 22-24 Gulfport, MS 157 32 27 2006 Mar 28-30 Mobile, AL 159 42 29 2007 April 23-25 Gainesville, FL 153 33 21 2008 Mar 24-26 Charleston, SC xx 33 22 2009 April 6-9 Oklahoma City, OK xx 19 11 2010 April 19-22 Little Rock, AR 91 25 16 2011 April 4-7 Galveston, TX 95 32 17 2012 April 16-18 Nashville, TN 66 20 9 2013 April 8-11 Virginia Beach, VA 70 20 11 2014 May 5-8 Palm Springs, CA 98 20 10 2015 April 6-8 New Orleans, LA 64 21 8 2016 May 23-25 Alburquerque, NM 60-ish 27(8+19)* 0 2017 May 16-18 Mobile, AL 69 20 7

* these meetings were held in conjunction with other meetings and presentation numbers indicate total IFA/ant presentation (symposium number + regular IFA meeting number)

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USDA Imported Fire Ant Quarantine History

Anne-Marie Callcott USDA-APHIS-PPQ

Caveat: this is an overview of information that I could get my hands on - gleaned from documents, manuals, and the Code of Federal Regulations (7CFR 301.81); not a full review of all publications about the IFA Quarantine There are currently 2 main laws that govern the Federal Imported Fire Ant Quarantine

• Plant Quarantine Act originally enacted in 1912 (7 U.S.C. 151 et seq.) o gave the authority “to regulate the importation of nursery stock and other plants

and plant products; to enable the Secretary of Agriculture to establish and maintain quarantine districts for plant diseases and insect pests; to permit and regulate the movement of fruits, plants, and vegetables therefrom and for other purposes.”

• Plant Protection Act of 2000 (7 U.S.C. 7701 et seq.) o consolidated authorities, including the Plant Quarantine Act, the Federal Plant

Pest Act and the Federal Noxious Weed Act of 1974 o “The Secretary may prohibit or restrict the importation, entry, exportation, or

movement in interstate commerce of any plant, plant product, biological control organism, noxious weed, article, or means of conveyance, if the Secretary determines that the prohibition or restriction is necessary to prevent the introduction into the United States or the dissemination of a plant pest or noxious weed within the United States.”

Prior to the Federal IFA Quarantine IFA were spreading throughout the southeast, slowly at first and then in the 1950s, when the suburbs came into being, and the nursery industry expanded, more quickly as humans helped spread the ants.

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How the Federal IFA Quarantine came into being; • 1957 – Southern Plant Board adopted resolution regarding cooperative control and

eradication program. National Association of Commissioners, Secretaries, and Directors of Agriculture supported SPB resolution.

• 1957 – USDA submitted an eradication plan to Congress • 1957 – Fall: first treatments applied in cooperative program. (2 lbs heptachlor/A – 20 lbs

of 10% granules) • 1958 – Federal quarantine invoked in eight southern states (7CFR 301.81)

o Alabama: 32 full counties and 9 partials o Arkansas: 1 partial county o Florida: 3 full and 10 partial o Georgia: 3 full and 1 partial o Louisiana: 21 full and 9 partial o Mississippi: 27 full and 17 partial o South Carolina: 2 partial o Texas: 6 full

From 1957-1976, USDA was involved with both:

1. Eradication program 2. Quarantine program

When the eradication program ended the quarantine program changed its mission from eradication to suppression to “slow the spread” Eradication era of the Federal IFA Program:

• 1957 – USDA-ARS-Plant Pest Control Division initiated eradication o heptachlor applied aerially

• 1962 – Mirex replaced heptachlor • 1962 – Silent Spring published • 1964-1965 – federal funding decreased

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• 1966 – federal funding increased • 1967 – USDA initiated new eradication program • 1970 – Mirex uses restricted • 1971 – Mirex registration cancelled and after court battle banned in 1978 • 1971 – APHIS established with regulatory mission for animals and plants • 1972 – USDA eradication efforts stopped • 1972-1978 – USDA provided some control to highly infested areas with states

During this time, USDA treated a vast amount of acreage, but the total is unknown. Many acres were treated multiple times. The following information is from a document in USDA files but its accuracy and provenance is unknown. For detailed information on the IFA eradication program read: “The Fire Ant Wars” by Joshua Blu Buhs.

Total Acreage Treated (accuracy unknown) FY 1958 449,553 FY 1959 931,981 FY 1960 892,521 FY 1961 452,834 FY 1962 974,104 FY 1963 1,857,994 FY 1964 2,347,681 FY 1965 3,672,040 FY 1966 6,128,542 FY 1967 10,247,043 FY 1968 12,833,187 FY 1969 11,121,521 FY 1970 14,508,107 FY 1971 11,609,907 FY 1972 11,065,153 FY 1973 14,184,017 FY 1974 13,599,628 FY 1975 12,679,181 FY 1976 & TQ 5,102,533

Over the years, the Federal IFA Quarantine program changed from eradication to suppression to “slow the spread”. The current (2017) mission of quarantine:

• To prevent the artificial spread of imported fire ants (S. invicta, S. richteri, and their hybrid) from where they are to where they aren’t – but could establish

o Establish a quarantine area o Regulate known pathways for IFA movement (nursery stock, hay, soil, bee

equipment, and anything else that can move fire ants)

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Regulatory/Treatment Era of the Federal IFA Program: USDA Players: 1958 – ARS Gulfport Methods Development Lab opened under ARS Plant Pest Control Division

and conducted methods development on eradication techniques 1958-1970 – no apparent methods development on regulatory treatments and dieldren and

heptachlor were used to certify sod and other plant material 1971 – APHIS established (regulatory section; ARS is research section)

• Gulfport Methods Development Lab moved to APHIS-PPQ o Reorganization cut staff and changed mission from eradication to control o Mission was to develop new and improved methods of controlling IFA

• ARS IFA Lab in Gulfport continued at a reduced level 1981 – APHIS Gulfport Methods Development lab realigned and became the Imported Fire Ant

Station • Mission was to develop quarantine treatments and areawide suppression techniques

1983 – ARS IFA Lab at Gulfport closed and moved to Gainesville, FL facility 2012 – APHIS-PPQ IFA Lab closed and quarantine methods development work outsourced Regulated Areas: 1962 – began using “generally infested area” and “eradication area” 1963 – eradication area defined as where cooperative treatments taking place 1969 – changed “eradication area” to “suppressive area” 1993 – removed “generally infested area” and “suppressive area” and just had a list of

quarantined area Regulated Articles: 1963 – Regulated articles:

• soil and unprocessed sand and gravel, separately or with other things • forest, field or nursery-grown woody or herbaceous plants with soil attached • plants in pots or containers, grass sod • unmanufactured forest products, such as stump wood or timbers if soil is attached • and any other products or articles that may be specifically determined likely to spread the

imported fire ant 2017 – Regulated articles:

• IFA queens and reproducing colonies of IFA • Soil, separately or with other articles, except potting soil that is shipped in original

containers in which the soil was placed after commercial preparation • Baled hay and baled straw stored in direct contact with the ground • Plants and sod with roots and soil attached, except plants maintained indoors in a home or

office environment and not for sale • Used soil-moving equipment, unless removed of all noncompacted soil; and • Any other article or means of conveyance when:

o An inspector determines that it presents a risk of spread of the imported fire ant due to its proximity to an infestation of the imported fire ant; and

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o The person in possession of the product, article, or means of conveyance has been notified that it is regulated under this subpart.

Certificates and Permits: 1960s (don’t know exact dates):

• Certificates may be issued for movement of regulated articles (paraphrased) under any one of the following:

1. Not been exposed to infestation 2. Examined by inspector and found free of infestation 3. Treated under observation of inspector and in accordance by him from

administratively authorized procedures know to be effective 4. Grown, produced, stored or handled in such a manner that no infestation would be

transmitted 2017 – little change

• Certificates may be issued for movement of regulated articles (paraphrased) under any one of the following

i. Is free of an imported fire ant infestation, based on his or her visual examination of the article;

ii. Has been grown, produced, manufactured, stored, or handled in a manner that would prevent infestation or destroy all life stages of the imported fire ant;

iii. Has been treated in accordance with part 305 of this chapter Treatments: 1958-1970

• Most treatments were dieldren and heptachlor 1975 Authorized Chemicals

• Fumigants o Eithylene dibromide-chlordane #431 (20%EDB; 10% chlordane) o Methyl bromide

• Insecticides o Chlordane o Mirex

1985 – Authorized Chemicals

• Fumigants o Ethylene dibromide #434 (2½ % EDB) o Methyl bromide

• Insecticides o Carbaryl o Chlordane (existing stock can be used for B&B, containers, sod per label) o Chlorpyrifos

• Examples of 1985 treatments o Hay and straw

If stacked in direct contact with ground bales above first layer can move without further treatment

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Methyl bromide if in direct contact with ground o Plants – balled or in container

Chlorpyrifos liquid: dip or drench 2X/day for 3 consecutive days Carbaryl (section 18 in limited states): one time drench for containers only Ethylene dibromide #434: inject into rootball (<36” diameter)

o Soil-Potting and Bench (containers) Chlorpyrifos granular: incorporation at ounces/cubic yard material with 24

month certification Methyl bromide: fumigate soil per schedule

o Sod Chlorpyrifos granular: broadcast at 4 or 6 lb ai/acre

1986 – EDB removed from list of approved chemicals (not sure when MeBr removed) 1988 – bait + chlorpyrifos approved for field grown stock (fenoxycarb) and, diazinon added

as 24(c) treatment for blueberries 1992 – hydramethylnon bait added to approved list 1992 – bifenthrin liquid approved for container drench treatment: first use of ppm dose rates 1993 – bifenthrin granular approved for container incorporation 1994 – tefluthrin granular approved for container incorporation 1999 – chlorpyrifos granular incorporation treatments removed for containers 1999 – pyriproxifen bait added 2002 – fipronil approved for use on containers and grass sod (manufacturer does not make label

for containers) 2003 – methoprene bait added 2013 – bifenthrin approved for use as B&B dip and on grass sod 2013 – revised B&B drench language and application to treat twice in one day and rotate/flip

rootball between applications 2015-2016 – abamectin and metaflumizone baits added 2017 Authorized insecticides

• Abamectin - bait • Bifenthrin • Chlorpyrifos • Diazinon (section 24(c) only for blueberries – none available) • Fenoxycarb - bait • Fipronil • Hydramethylnon - bait • Metaflumizone - bait • Methoprene - bait • Pyriproxyfen - bait • Tefluthrin (no label available)

• Examples of 2017 Treatments

o Nursery Stock – Containers Incorporation (bifenthrin, fipronil, tefluthrin)

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Drench (bifenthrin, chlorpyrifos) Dip/Immersion (bifenthrin, chlorpyrifos)

o Nursery Stock – Balled or Field Grown Balled (B&B – postharvest) Drench (chlorpyrifos) Dip/Immersion (bifenthrin, chlorpyrifos) Field Grown (preharvest)

• Bait + contact insecticide (several baits + chlorpyrifos) o Grass Sod

Broadcast insecticide (bifenthrin, fipronil) o Hay and straw

Baled hay and straw stored in direct contact with the ground is ineligible for movement from the quarantined area to an area outside the quarantine, unless inspected, found free of IFA, and issued a certificate

o Greenhouse grown plants Certifiable without treatment if inspector determines that the greenhouse is

constructed of fiberglass, glass, or plastic in such a way that IFA is physically excluded and cannot become established

o Soil – Bulk Heated

o Soil – samples Heated or frozen

USDA-APHIS webpage has program aids and other info for growers impacted by the quarantine including a new interactive quarantine map that allows viewer to type in an address to see if it is in the regulated area or to zoom into street level to see quarantine lines. https://www.aphis.usda.gov/aphis/ourfocus/planthealth/plant-pest-and-disease-programs/pests-and-diseases/imported-fire-ants/ct_imported_fire_ants

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Biosecurity and Surveillance of Ants in Australia

Antonette Walford Department of Agriculture and Water Resources, Melbourne, Australia.

www.agriculture.gov.au/biosecurity antonette.walford@agriculture.gov.au

The Department of Agriculture and Water Resources protects Australia from exotic, invasive species to protect agriculture, environment and human health. This is achieved through inspection of imported goods and surveillance at properties where imported goods are received including international air and seaports. Hundreds of invasive and tramp ants are intercepted and identified at Australia’s borders, including fire ants (Solenopsis invicta). Preventing an incursion or establishment of invasive species is considered valuable as early detection yields a significantly higher return on investment than the costs associated with control or eradication for an established population. The department began delivery of a National Border Surveillance (NBS) program from November 2016. One aim of this program is to survey sites associated with imported goods for invasive ants, in order to achieve early detection of incursions. The department continues to build capability and resources to ensure competence for rapid identification and recognition of exotic and endemic species. Part of this development included my 2017 trip to the USA to work on relevant diagnostic challenges, collect species not found in Australia and to build professional networks. It also allowed discussion and observation of current surveillance techniques with experienced professionals to identify potential improvements.

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Imported Fire Ant Worker Size as Relates to Species/Hybrid and Geographic Distribution in Tennessee

Manoj Pandey,1 Jason Oliver, 1 Karla Addesso, 1 Steven Valles, 2 Reginald Archer, 1

and Anne-Marie Callcott3

1 Tennessee State University, College of Agriculture, Human, and Natural Sciences, 3500 John A. Merritt Boulevard, Nashville, TN.

2United States Department of Agriculture - Agricultural Research Service, Center for Medical, Agricultural and Veterinary Entomology (CMAVE), 1700 SW 23rd Drive, Gainesville, FL.

3United States Department of Agriculture – Animal, Plant Health Inspection Service – Plant Protection and Quarantine, Biloxi Station, 1815 Popps Ferry Road, Biloxi, MS.

Imported fire ants (IFA), Solenopsis spp. (Hymenoptera: Formicidae) have continued to expand their range since entering Tennessee in 1987. Past surveys have indicated that red (Solenopsis invicta Buren) and black (Solenopsis richteri Forel) IFA and their hybrid, S. invicta × S. richteri constitute about 2.3, 43.2, and 54.5% of the Tennessee IFA population, respectively (Oliver et al. 2009). These ants are polymorphic and exhibit a range of worker ant sizes. The IFA species and worker size are relevant to size-dependent biological control agents like Pseudacteon spp., particularly in states like Tennessee with mixed populations of black, red and hybrid IFA. The objective of this research was to determine the relationship of IFA species and their hybrids with worker ant size, and the geographic distribution of IFA species and their hybrids in Tennessee. Imported fire ant workers were collected from 440 colonies in 62 infested Tennessee counties south of the Federal Imported Fire Ant Quarantine (TDA 2017) from late July to early October in 2015 and 2016. Counties were divided into 16.1 × 16.1 km quadrants and workers from one IFA colony were collected near the center of each grid quadrant. Each sample location was mapped with global positioning units (GPS) (LandMark Spatial Systems, LLC, Starkville, MS) that included either a Flint S Series F4 Device hand-held unit running SOLO Forest for Windows CE version 4.0.7 or a Nomad Data Collector (Tripod Data Systems, Inc., Corvallis, OR) running the same software in version 4.0.5. The Flint unit had an internal antenna, whereas the Nomad unit was connected by Bluetooth to a Hemisphere GPS (Crescent A100 Model and version 6.8 /2.6K) which received Wide Area Augmentation System (WAAS) corrected signal. Worker ants were sampled by digging into the IFA mound and then allowing ants to crawl up a wooden board (90 by 3.5 by 2.0 cm). Ants were then knocked into an 18.9 L bucket, which had the rim treated with Insect-a-Slip Insect Barrier – Fluon (Catalog number 2871; BioQuip Products Rancho Dominguez, CA) or baby powder to prevent ant escape. Cuticular hydrocarbons and venom alkaloids of workers were collected via hexane extraction from sampled colonies and analyzed by gas chromatography and mass spectrometry. Cuticular hydrocarbon and venom alkaloid profiles were used to determine IFA species (i.e., black or red IFA) or their hybrid using procedures developed by Vander Meer et al. (1985) and Ross et al. (1987). Although not assessed molecularly, it was assumed all of the black and hybrid IFA colonies in Tennessee were monogyne based on low incidence of polygyny in hybrid and black IFA (Menzel et al. 2008) and the polymorphism of worker sizes observed in all Tennessee colonies. Thirteen red IFA colonies from a Gainesville, FL monogyne site were collected for worker size comparisons with Tennessee black and hybrid colonies, due to the low incidence of red IFA in Tennessee samples.

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Among the 440 Tennessee samples, a sub-sample of colony collections were randomly selected, the head capsules of 50 worker ants from each colony were photographed under AxioCam ERc Microscope Camera (ZEISS, Jena, Germany), and head size was measured with Image J software (National Institute of Health, New York, NY). Statistical analysis was performed to determine whether ant species/ hybrid status and geographical distribution contributed to worker size. Average Nearest Neighbor Analysis was used to determine the geographic distribution pattern of colonies with similar average worker ant size in ArcGIS 10.3.1 (ESRI, Redlands, CA). The study results indicated similar patterns of head size between black and hybrid IFA colonies. The average worker size of red IFA collected from FL was smaller than black or hybrid IFA. The colonies with similar head size were randomly distributed geographically within Tennessee, and therefore, regional location had no detectable effect on average head size in colonies. Also, the mean head capsule width (mm) of the colonies was not related to elevation. Among the 440 colonies collected, 336 (76.3%), 101 (23.0%), and 3 (0.7%) were identified as hybrid, black, and red IFA, respectively. The eastern part of the state was mostly dominated by hybrid IFA with exception of a couple of red IFA colonies. The middle part of the state was a mixture of black IFA and hybrid IFA with black IFA frequency increasing in the far western counties of the state. Three red IFA colonies in total were found in Davidson, Jefferson, and McMinn counties. There appears to be an increase in the proportion of hybrid IFA in the Tennessee IFA population compared to the previous study conducted a decade ago (Oliver et al. 2009). We thank USDA-APHIS-PPQ for providing partial funding of this study (Cooperative Agreement Award 15-8130-0523-CA). Literature Cited

Menzel, T. O., D. C. Cross, J. Chen, M. A. Caprio, and T. E. Nebeker. 2008. A survey of imported fire ant (Hymenoptera: Formicidae), species and social forms across four counties in east-central Mississippi. Midsouth Entomologist 1: 3−10.

Oliver, J. B., R. K. Vander Meer, S. A. Ochieng, N. N. Youssef, E. Pantaleoni, F. A. Mrema, K. M. Vail, J. P. Parkman, S. M. Valles, W. G. Haun, and S. Powell. 2009. Statewide survey of imported fire ant (Hymenoptera: Formicidae) populations in Tennessee. Journal of Entomological Science 44: 149−157.

Ross, K. G., R. K. Vander Meer, D. J. Fletcher, and E. L. Vargo. 1987. Biochemical phenotypic and genetic studies of two introduced fire ants and their hybrid (Hymenoptera: Formicidae). Evolution 41: 280−293.

Vander Meer, R. K., C. S. Lofgren, and F. M. Alvarez. 1985. Biochemical evidence for hybridization in fire ants. Florida Entomologist 68: 501−506.

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“Fighting Texas’ Fire Ants: The Team Approach” 15 Years and Counting – Review of the Lago Santa Fe Fire Ant Project

Paul R. Nester, Extension Program Specialist – IPM

Texas A&M AgriLife Extension Service, Houston, TX Managing the red imported fire ant, Solenopsis invicta Buren (Hymenoptera: Formicidae) through broadcast applications of fire ant bait products has been demonstrated to dramatically reduce the cost, insecticide use, maintain control of fire ants, and help eliminate problems caused by the fire ant (Riggs et.al, 2002). The Lago Santa Fe Fire Ant Project is a good example of a successful community-wide fire ant management program where the bi-annual broadcasting of a fire ant bait product (Extinguish® Plus) has resulted in continued suppression of a fire ant population. Lago Santa Fe is a Private Lake Community, situated 25 miles, South/Southwest of Houston, TX, in Galveston Co. The Community consists of 100 acres with four, ½ mile long X 200 ft wide lakes, with 48, one acre lots that border the lakes and 17 lots that do not border the lakes (Nester et.al, 2003).

Historical Perspective, 2001 – 2002. In 2001, Lago Santa Fe (Figure 1) was chosen as the site to host the 2002 National Water Ski Championships and the 2002 U.S. Open Water Ski Championships. This meant that in August of 2002, 1000 participants from all over the U.S. and the World, with up to 4,000 spectators would invade this 100 acre area for 7 days. Initial fire ant mound activity evaluations showed over 160 large (> 12” diameter) active fire ant mounds per acre. The Lago Santa Fe Community realized they needed a coordinated approach to manage these pests. The Texas A&M AgriLife Extension Service decided to work with the residents of Lago Santa Fe and develop an annual repeatable process for the management of the fire ants in this community. The Lago Santa Fe Fire Ant Project gave The Texas A&M AgriLife Extension Service a chance to “showcase” various ideas, practices, products, and product uses developed by the Texas Imported Fire Ant Research and Management Plan which included:

1) Evaluation of the effectiveness of the "hopper blend" treatment (50:50 blend of

hydramethylnon plus s-methoprene ant bait later released as Extinguish® Plus) at 1.5 lb. blended product/acre.

2) Demonstration of application methods such as the truck-mountable industrial "bait blower"; and the ATV mounted Herd Broadcast Spreader.

3) Demonstration of the practicality of scheduling fire ant bait treatments prior to events to reach a goal of maximum control such as the 2002 and 2003 National Water Ski Championships, and U.S. Open Water Ski Championships being hosted by the Lago Santa Fe Community

4) Demonstration of how coordinating the efforts of the entire community resulted in an efficient process for managing fire ants over the Lago Santa Fe property.

Fire Ant mound activity counts showed that after a single spring 2002 hopper blend treatment, fire ant activity was reduced 85% in the community of Lago Santa Fe before the scheduled 2002 water ski events. Full report documenting this project can be found at: http://www.extension.org/sites/default/files/w/0/03/2003_IFA_Conference_Proceedings.pdf. A video documenting this effort was created, YRS31blpT-M, and can be found on YouTube.com.

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2002 – 2004 ongoing program treatments: This community was also chosen to host these same events in the summer of 2003. In the succeeding years after the successfully implementation of the Lago Santa Fe fire ant project the community reduced the highs and lowered the lows of the yearly fluctuations in fire ant numbers. After an additional fall 2002 and spring 2003 hopper blend treatment, fire ant mound activity was down 95%, before the scheduled 2003 water ski events. The community has continued to manage their fire ants. After 5 scheduled baiting events (April 2002 through May 2004) the fire ant mound activity reduction remained above 90% when compared to initial fire ant activity recorded in the spring of 2002.

2003 – 2008 ongoing program treatments: Lago Santa Fe residents continue their efforts to manage the fire ant with bi-annual applications of a fire ant bait product. Lago Santa Fe residents pay quarterly dues to the Property Owners Association (POA). Fire ant baiting is funded from the general dues just like mowing, lake dye, insurance, electricity etc. Fire ant bait is a line item in the budget that is approved annually by the Board of Directors (BOD). The most recent (2011) fire ant budget consists of seven 25 pound bags of fire ant bait, miscellaneous supplies and fuel for ATV on which a Herd G-77 Broadcast Seeder is mounted. It is estimated that each property owner will pay about $20 for fire ant management every 6 months for a total cost of $40/year/property. In a 2008, the Lago Santa Fe Fire Ant Project was re-visited and the residents surveyed as to their satisfaction of this project over the past years (Nester et.al, 2008). The residents (11 responses from 40 property owners, 28%) all responded (100%) that they felt the dollars spent by the POA for the Lago Santa Fe Fire Ant Project was cost-effective and all responded (100%) that they wanted it to continue. All (100%) ranked the project from good to excellent and 10 of the residents (91%) felt very positive about this fire ant baiting program being supported by the Texas AgriLife Extension Service. One of the residents was somewhat positive. One comment from a resident was, “Prior to moving to TX / Lago Santa Fe a few years ago, we had never lived anywhere that had a fire ant problem. We’ve only been exposed to the problem and solutions since living here but have heard numerous stories from neighbors as to how bad it was and how it is today. We certainly appreciate all the efforts made by Texas A&M AgriLife Extension Service and Jay and Lydia Gilbert”

2010 – 2011 program activities: Extenuating circumstances kept the original fire ant management organizers from coordinating the baiting event in the fall of 2010. Since other community members did not pick up the effort, the fall 2010 community-wide baiting event did not happen. As the year ended and the new (2011) year began, there was chatter in the community indicating that fire ant populations were increasing on the various properties, in numbers not seen in previous years (personal communication with Jay Gilbert, Lago Santa Fe resident, fall 2010). When communicating with Jay Gilbert, one of the original organizers of the Lago Santa Fe Fire Ant Project, it was decided to try and renew interest in the community-wide project by making a concerted effort in the spring of 2011 (10 years after the first fire ant baiting event) to make all residents aware of the importance of the bi-annual baiting by organizing an event and taking before and after observations of fire ant activity.

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Materials and Methods for 2011 follow-up evaluation. A date to spread fire ant bait was set. Initial fire ant mound activity observations were taken using the minimal disturbance method April 25, 2011from 0.25 acre circles from 6 properties within the Lago Santa Fe community (a reduction in plot numbers due to houses constructed on previously vacant lots). As in previous years, fire ant activity observations and ant mound counts were also taken from four 0.25 acre circles in an adjacent untreated pasture to serve as a control plot area. These same treated and untreated plot locations were checked 90 days later for fire ant mound activity. At the same time we conducted active ant mound counts in plots, we measured fire ant foraging activity by placing 20 hot dog food lures (0.25 inch thick slices of Bar-S Jumbo Franks) in the front yards of 20 of the interior properties that border the lakes. After 60 minutes, the total estimated number of fire ants per lure was recorded. Food lures were also used 90 days later to check for fire ant foraging activity. Fire ant bait product using the active ingredients hydramethylnon + s-methoprene (Extinguish® Plus Fire Ant Bait) was donated by Central Life Sciences. This is the only fire ant bait active ingredients that had been used on this property in previous years. This time the Lago Santa Fe Fire Ant Project gave The Texas A&M AgriLife Extension Service a chance to again:

1) Showcase The Community –Wide fire ant baiting concept promoted by the Texas

AgriLife Extension Service. 2) Demonstrate how coordinating the efforts of the entire community can result in an

efficient process for managing fire ants over the Lago Santa Fe property. 3) Evaluate if tolerance or resistance (behavioral or physiological) of the fire ants in this

community to the active ingredients hydramethylnon and s-methoprene was occurring.

Results and Discussion What started out to be a promising year for obtaining fire ant management data turned into a record setting year for Texas by having the hottest average temperature for June through August according to National Weather climatologists. The spring and summer of 2011 was extremely dry in south Texas. One would assume that the irrigation of lawns would attract fire ants. The lawns of Lago Santa Fe were no exception. Good fire ant activity was seen April 25, 2011. Of the 20 food lures, 19 (95%) of them had an average of 54 fire ants per lure. In April 2002 over 160 large (> 12” diameter) active fire ant mounds per acre were observed. The mounds (136/acre) observed in April 2011 were not overly large but they were present. Fire ant mound activity counts taken in an adjacent pasture also showed good activity at 112 active fire ant mounds per acre. The dry conditions did have an effect on mound survivability as seen by the 36% reduction in activity with no bait application in the adjacent pasture. Greater than 90% reduction of the fire ant mound activity was observed in the treated properties of Lago Santa Fe when compared to pre-treatment counts, and a 74% reduction in food lure hits was observed. Were the residents of Lago Santa Fe satisfied with the 2011 effort? An email from Jay Gilbert, one of the originators of the Lago Santa Fe Fire Ant Project said, “Fall baiting was in October

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2011 and was facilitated with 4 block captains. Utilizing block captains reduced the burden on the 2 of us and we are more willing to continue overseeing the semi-annual baiting. The Fall baiting appeared successful - as there were no complaints compared to lots of negative chatter last year. You will recall we skipped the fall baiting that year (2010) and the ants really were a problem. I have only spotted a couple of nuisance mounds following a 6.5" rain event Jan 9, 2012. We will go after them again in the Spring.” Block captains were chosen to help facilitate future fire ant management events. 2011 Summary - 10 years of community planned fire ant management:

• Sample size was reduced for 2011 counts due to construction on previously empty lots. • Despite drought, populations in untreated area (pastureland) remained unchanged,

statistically. • Percent reduction from April 25, 2011 to July 27, 2011 was 94.7% (versus 85.5%

reduction after initial treatment, April 18, 2002 to July 7, 2002), indicating that after annual multiple application, Extinguish® Plus continues to perform over 10 years with no indication of resistance by imported fire ant populations.

• The Community-Wide fire ant management concept as promoted by the Texas AgriLife Extension Service is still a viable solution to the management of the red imported fire ants in a community setting.

2017 follow-up of 15 years of a community planned fire ant management In the spring of 2017 Jay and Lydia Gilbert, longtime residents of the Lago Santa Fe water skiing community were contacted for a 15 year follow-up to the initiation of the original Lago Santa Fe Fire Ant Management Project started back in spring 2002. Jay outlined the most recent iteration of the long lasting Lago Santa Fe Fire Ant Project.

• Lago Santa Fe has had a viable fire ant management program since April 2002. • Since 2002 Lago Santa Fe residents have treated every fall and spring except for fall

2010. • The purchase of a fire ant bait product is a budgeted item for the Lago Santa Fe

Community Property Owners Association (POA). • One household organizes the distribution and co-ordinates volunteers for common areas

and absentee owner properties. They used a committee for a couple of years but it ended up being more trouble than it was worth.

• The hopper Blend of Amdro + Extinguish or the product Extinguish Plus (since 2004) has been exclusively used by the property owners of Lago Santa Fe as the first part of the 2 step approach.

• A Herd Broadcast Seeder mounted to an ATV was used up to 2012 when there were many absentee land owners, 38 out of 65 lots were developed in 2002 as opposed to 61 developed in 2017. It is now retired. (Figure 2)

• Now since all but 4 of the 65 lots are occupied, the POA purchases the Extinguish Plus fire ant bait product and it is divided up and given to the homeowners to treat properties they are assigned.

• Of the original 38 participants, 29 still reside at Lago Santa Fe.

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• The Lago Santa Fe Community mainly focus on the properties (both occupied and publicly shared) and do not treat the outside perimeter boundary area of the lakes since the cost and effort vs benefit didn't favor their continued treatment.

2017 Final Summary - 15 years of community planned fire ant management The Lago Santa Fe Water Skiing Community in Santa Fe, TX, is still treating for fire ants with a slightly altered management strategy.

• The Lago Santa Fe Community still supports the community-wide treatment concept for fire ants as planned back in 2002.

• The Community will still plan for a biannual community “baiting weekend.” • They have a property owner that will organize the biannual event. • The POA will purchase product for the community. • The product will be passed out to property owners. • Property owners will use handheld seeders to broadcast the fire ant bait product • Property owners will treat adjacent vacant lots and public areas. • The outside perimeter boundary area of the lakes will no longer be treated. • When new property owners move in they are told of the Lago Santa Fe video

(YRS31blpT-M) explaining why the POA stresses the need for the fire ant community-wide baiting.

• Community leaders have made fire ant baiting a required community activity • According to a Lago Santa Fe a POA Representative, and longtime resident Jay Gilbert,

Lago residents are very satisfied with the fire ant baiting program. According to Jay, “The baiting program has improved the outdoor experience for the entire community over the past 15 years. In fact since the Extinguish Plus product is now readily available online and in smaller 1.0 & 4.5 lb. containers. I have directed some non-Lago people to buy small containers of Extinguish Plus for their individual property so they can benefit from area wide control of fire ants.”

Acknowledgements The author want to thank the residents of Lago Santa Fe for allowing him access to their community over the past 15 years to document events and follow the effects of their fire ant management attempts. Special thanks go to Jay and Lydia Gilbert for their willingness to meet with the authors when necessary and help with fire ant activity evaluations. Central Life Sciences (Doug Van Gundy) is thanked for the generous donation of fire ant bait product when it was needed.

Literature cited: Nester, P.R., C.P. Bowen, B. M. Drees. 2003. The Lago Santa Fe Fire Ant Project: An example

of Community-wide imported fire ant management in Texas. 2003 proceedings of the Red Imported Fire Ant Conference.

Nester, P.R., C.P. Bowen, B. M. Drees. 2008. “Fighting Texas’ Fire Ants: the Team Approach” A six year review of the Lago Santa Fe Fire Ant Project. Pg 36-42. Urban IPM Summary

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report. Texas AgriLife Extension Service posted at: http://fireant.tamu.edu/research/projects/pdf/ipmmanual08a1.pdf

Riggs, N. L., L. Lennon, C. L. Barr, B. M. Drees, S. Cummings, and C. Lard. 2002. Community-wide red imported fire ant programs in Texas (B. M. Drees, ed.). Southwestern Entomologist Supplement No. 25:31-42 posted at: http://www.extension.org/sites/default/files/w/0/03/2003_IFA_Conference_Proceedings.pdf.

Figure 1. Lago Santa Fe subdividion, Galvestion Co., TX, site of a community wide fire ant management program, 2002-2012.

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Figure 2. Lago Santa Fe subdivision, Galveston Co., TX, site of a community wide fire ant management program, 2002-2017. View of development over the past 15 years.

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Evaluation and Review of the Scotts® Electric Powered Handseeder – the Wizz™

Timothy S. Davis, PhD

University of Georgia Extension Agent – Chatham County Handseeders operated with a hand crank have been used for years in smaller yards and plot for the broadcast application of fire ant bait. The adjustments allow the bait to be spread at the appropriate rates. Through the years several models have been available. The Scotts® Handy Green II™ (pictured below) has been a much used model. The wrist loop allows for applications as large as one acre with minimal fatigue impacting the application rate. Recently, Scotts® introduced the Wizz™, a battery powered version of their handseeder. This paper is an evaluation and review of the Wizz™ for the purpose of fire ant bait applications.

The Wizz™ handseeder was purchased from a big box store off the shelf. Amdro Fire Ant Bait was also purchased off the shelf at a big box store to simulate the conditions that a typical home owner would encounter with the product. The tests were NOT replicated using multiple baits or multiple spreaders and is not intended to be research. It is intended to review the product for the suitability of spreading fire ant bait in a homeowner situation. A side by side comparison shows that the Wizz™ is slightly larger and heavier than the Handy Green II and lacks the wrist loop. Since the Wizz™ is powered by an electric motor it can be held in a different posture and the need for the wrist loop is eliminated.

The back side of the label has operating instructions and settings for some common products that might be spread using the Wizz™ – all Scotts® products. It does not have any calibration settings for any fire ant baits included.

The Wizz™ is powered by four AA batteries that are supplied with the original purpose. They are changeable. One complaint is that screw to open the battery compartment was very small and difficult to open with a standard Philips head screwdriver. Battery life was very good and ran for over two hours continuously without slowing down before the test was terminated

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The calibration control dial on the Wizz™ had more settings and allowed for finer tuning between settings. Fire ant bait requires a minimum of 1/8 in opening. The minimum setting on the Wizz™ was smaller than 1/8 in. When opened to the highest setting we were able to achieve the satisfying and over applied yellow cloud. Proper application rates were relatively easy to achieve at the 2.75 to 3 setting on the particular spreader used in this review. A full acre lot was treated with 1.5 lbs of Amdro Fire Ant bait. Swath width with the Wizz™ averaged 10ft on as measured after 10 passes over tar paper. This is more than sufficient for

an appropriate application of fire ant bait. The trigger pull was stiff and difficult to hold leading to fatigue during the application. Fortunately the Wizz™ has a trigger lock mechanism that keeps the spreader running without the need to constantly squeeze the trigger. This feature significantly reduced fatigue and contributed to a more even spread of the fire ant bait across the test plot. Hand crank speed with the Handy Green II impacted the product dispersal rate. The limitations of an individual person to crank at a consistent speed has always introduced application variability. The battery powered spreader ran at a consistent speed which reduced the application variability introduced by the spinner and agitator speed. Conclusions: The Wizz™ is more adjustable than the old product. The battery power reduces variability over the hand cranked unit. The motor locking feature reduced overall fatigue during the application. Based on the performance of the Wizz™ it can be recommended for small applications of less than 1 acre in size.

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Geographic Incidence of Kneallhazia solenopsae and Solenopsis invicta Viruses (SINV-1, -2, and -3) in Imported Fire Ant Species and their Hybrid in

Tennessee Jason Oliver,1 Karla Addesso,1 Steven Valles,2 Nadeer Youssef,1 Manoj Pandey,1 Paul O'Neal,1 Joshua Basham,1 Joseph Lampley,1 Debbie Eskandarnia,1 Garrett Roper,1 Megan Patton,1 Steve

Powell,3 and Anne-Marie Callcott4

1 Tennessee State University, College of Agriculture, Human, and Natural Sciences, Otis L.

Floyd Nursery Research Center, 472 Cadillac Lane, McMinnville, TN. 2United States Department of Agriculture - Agricultural Research Service, Center for Medical,

Agricultural and Veterinary Entomology (CMAVE), 1700 SW 23rd Drive, Gainesville, FL. 3Tennessee Department of Agriculture, Plant Certification Section, Ellington Agricultural

Center, 440 Hogan Road, Porter Building, Nashville, TN. United States Department of Agriculture – Animal, Plant Health Inspection Service – Plant

Protection and Quarantine, Biloxi Station, 1815 Popps Ferry Road, Biloxi, MS.

Imported fire ant (Solenopsis spp.) (IFA) pathogens like Kneallhazia solenopsae Knell Allen Hazard and Solenopsis invicta viruses (SINV-1, -2, and -3) can impact colony reproduction and survival (Valles et al. 2013, Porter et al. 2013). Pathogens are widespread in the southern parts of the red IFA (Solenopsis invicta Buren) range, but knowledge of pathogen distribution is limited in the northern range areas dominated by black IFA (Solenopsis richteri Forel) and the S. invicta × S. richteri hybrid IFA (Valles et al. 2007). In Tennessee, past surveys have indicated the IFA complex is primarily black IFA in west Tennessee and hybrid IFA in middle and east Tennessee with red IFA being rare (Oliver et al. 2009). The objective of this study was to determine the incidence and seasonality of Kneallhazia and SINV viruses in Tennessee. The goal of the project was to assess the need for redistribution efforts of IFA pathogens in the northern parts of the IFA range. The IFA infested areas south of the Federal IFA Quarantine line in Tennessee were divided into 16.1 by 16.1 km (10 by 10 mile) survey grids. A single IFA colony was sampled near the middle of each survey grid during summer (late July to early October in 2015 and 2016) and during the winter (January to early March 2016). If colonies could not be located after about a 30 minute search of the survey grid center, then the grid was not sampled. Because of potential variation in pathogen seasonal occurrence (Valles et al. 2010), samples were collected in both summer and winter. During the summer, all survey grids were sampled when possible (n= 440), but in the winter when temperatures were lower and colonies potentially harder to sample, every other grid was surveyed (n=227) (i.e., checker board pattern). Worker ants were collected by using a trowel to open the mound and then placing a wooden stick on the surface to let the ants crawl up. Ants were then tapped into an 11.4 liter bucket treated with an escape-prevention barrier of either Insect-a-Slip Insect Barrier (fluoropolymer resin [PTFE-30]) (BioQuip Products, Rancho Dominguez, CA) or baby powder. About 100 worker ants were removed and placed into scintillation vials with 95% ethyl alcohol (A.C.S. spectrophotometric grade, Item No. 61511-0040, CAS 64-17-5; Arcos Organics – Fisher Scientific, Fair Lawn, NJ). The ant samples in ethanol were analyzed at the CMAVE facility for the presence of SINV-1, -2, or -3 or K. solenopsae using molecular procedures (Valles et al. 2002, 2004, 2009). An additional ~50

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workers were collected live and were kept in a cooler with ice packs until they could be immersed in n-hexane Chromasolv® for High Performance Liquid Chromatography (Sigma-Aldrich, Saint Louis, MO) for cuticular hydrocarbon and venom alkaloid analysis with a Shimadzu QP-2010 Gas Chromatograph - Mass Spectrometer (GC-MS) (Shimadzu, Kyoto, Japan) (Ross et al. 1987; Vander Meer et al. 1985, 1989). An additional 1,000+ workers were collected and subsequently stored in a -80oC freezer for any additional needs. Each site location was mapped with global positioning units (GPS) (LandMark Spatial Systems, LLC, Starkville, MS) that included either a Flint S Series F4 Device hand-held unit running SOLO Forest for Windows CE version 4.0.7 or a Nomad Data Collector (Tripod Data Systems, Inc.) running the same software in version 4.0.5. The Flint unit had an internal antenna, whereas the Nomad unit was connected by Bluetooth to a Hemisphere GPS (Crescent A100 Model and version 6.8 /2.6K) which received Wide Area Augmentation System (WAAS) corrected signal. Pathogen incidence in Tennessee was very low. During summer months, 6.1, 0.2, 1.4, and 1.1% of the colonies were infected with SINV-1, SINV-2, SINV-3 and K. solenopsae, respectively, while during the winter only SINV-1 and K. solenopsae were detected in 0.4 and 0.9% of the colonies, respectively. The SINV-1 virus occurred statewide, SINV-2 was found at only one site in Rutherford County, SINV-3 was found in one southern and three east Tennessee counties, and K. solenopsae was mostly restricted to west Tennessee. The percentage of infected colonies in Tennessee are much lower than the 73 to 93% infection rates reported for Florida monogyne and polygyne colonies, respectively (Allen et al. 2011). In addition, no colonies in Tennessee were found with multiple pathogen infections, which occurred in 13 and 33% of Florida monogyne and polygyne colonies, respectively (Allen et al. 2011). Only four red IFA colonies were found during the survey in one west, one middle, and two east Tennessee counties. All other IFA samples in east Tennessee were hybrid IFA, in middle Tennessee most of the IFA were hybrid, and west Tennessee had a mixture of hybrid and black IFA. Hybrid IFA had increased in prevalence in western Tennessee counties compared to a previous survey done during 2004 to 2006 (Oliver et al. 2009). Kneallhazia and SINV-1 were detected in black IFA, while all four pathogen types were found in hybrid IFA. Detailed data and distribution maps will be published in a future journal article. Fig. 1. Clockwise from left, topping a fire ant mound with a trowel, collecting fire ant workers with a wooden board, fire ants in a 11.4 liter bucket lined with an escape-prevention barrier, and examples of some fire ant collecting sites near railroad tracks and roadsides.

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Literature Cited Allen, C., S. M. Valles, and C. A. Strong. 2011. Multiple virus infections occur in individual polygyne and monogyne Solenopsis invicta ants. Journal of Invertebrate Pathology 107: 107−111. Oliver, J. B., R. K. Vander Meer, S. A Ochieng, N. N. Youssef, E. Pantaleoni, F. A. Mrema, K. M. Vail, J. P. Parkman, S. M. Valles, W. G. Haun, and S. Powell. 2009. Statewide survey of imported fire ant (Hymenoptera: Formicidae) populations in Tennessee. Journal of Entomological Science 44: 149−157. Porter, S. D., S. M. Valles, and D. H. Oi. 2013. Host specificity and colony impacts of the fire ant pathogen, Solenopsis invicta virus 3. Journal of Invertebrate Pathology 114: 1−6. Ross, K. G., R. K. Vander Meer, D. J. Fletcher, and E. L. Vargo. 1987. Biochemical phenotypic and genetic studies of two introduced fire ants and their hybrid (Hymenoptera: Formicidae). Evolution 41: 280−293. Valles, S. M., D. H. Oi, O. P. Perera, and D. F. Williams. 2002. Detection of Thelohania solenopsae (Microsporidia: Thelohaniidae) in Solenopsis invicta (Hymenoptera: Formicidae) by multiplex PCR. Journal of Invertebrate Pathology 81: 196−201. Valles, S. M., C. A. Strong, P. M. Dang, W. B. Hunter, R. M. Pereira, D. H. Oi, A. M. Shapiro, and D. F. Williams. 2004. A picorna-like virus from the red imported fire ant, Solenopsis invicta: initial discovery, genome sequence, and characterization. Virology 328: 151−157. Valles, S. M., C. A. Strong, D. H. Oi, S. D. Porter, R. M. Pereira, R. K. Vander Meer, Y. Hashimoto, L. M. Hooper-Bùi, H. Sánchez-Arroyo, T. Davis, V. Karpakakunjaram, K. M. Vail, L. C. Graham, J. A. Briano, L. A. Calcaterra, L. E. Gilbert, R. Ward, K. Ward, J. B. Oliver, G. Taniguchi, and D. C. Thompson. 2007. Phenology, distribution, and host specificity of Solenopsis invicta virus-1. Journal of Invertebrate Pathology 96: 18−27. Valles, S. M., L. Varone, L. Ramírez, and J. Briano. 2009. Multiplex detection of Solenopsis invicta viruses -1, -2, and -3. J. Virological Methods 162: 276−279. Valles, S. M., D. H. Oi, S. D. Porter. 2010. Seasonal variation and the co-occurrence of four pathogens and a group of parasites among monogyne and polygyne fire ant colonies. Biological Control 54: 342−348. Valles, S. M., S. D. Porter, M. Choi, and D. H. Oi. 2013. Successful transmission of Solenopsis invicta virus 3 to Solenopsis invicta fire and colonies in oil, sugar, and cricket bait formulations. Journal of Invertebrate Pathology 113: 198−204. Vander Meer, R. K., C. S. Lofgren, and F. M. Alvarez. 1985. Biochemical evidence for hybridization in fire ants. Florida Entomologist 68: 501−506. Vander Meer, R. K. and C. S. Lofgren. 1989. Biochemical and behavioral evidence for hybridization between fire ants, Solenopsis invicta and Solenopsis richteri (Hymenoptera: Formicidae). Journal of Chemical Ecology 15: 1757−1765.

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Introduction and Establishment of Fire Ant Biocontrol Agents in the Coachella Valley of California

David Oi1, Steven Valles1, Sanford Porter1,

Christopher Cavanaugh2, Greg White2, 3, and Jennifer Henke2

1USDA-ARS Center for Medical, Agricultural, and Veterinary Entomology 1600 SW 23rd Drive, Gainesville, Florida 32608

2Coachella Valley Mosquito and Vector Control District 43-420 Trader Place, Indio, CA 92201

3(currently: Salt Lake City Mosquito Abatement District, Salt Lake City, Utah 84116) A collaborative project between the USDA-ARS Center for Medical, Agricultural, and Veterinary Entomology and the Coachella Valley Mosquito and Vector Control District (CVMVCD) was initiated in 2014 to establish biological control agents of the red imported fire ant, Solenopsis invicta, in the Coachella Valley of Riverside County, California. The following biocontrol agents were released: the microsporidian fire ant pathogen Kneallhazia solenopsae, Solenopsis invicta virus 3 (SINV-3), and two species of fire ant decapitating phorid flies. Red imported fire ants (or ‘fire ants’ hereafter) generally inhabit the irrigated, urban landscapes, in the desert climate of the Coachella Valley. The objective of this project was to determine if the biocontrols could survive in the hot, irrigated habitat. Permits to release K. solenopsae and phorid flies were issued by the USDA Animal and Plant Health Inspection Service with concurrence from the California Dept. of Food and Agriculture. A permit to release SINV-3, a domestically isolated insect virus, was not needed under current policy. Four sprinkler irrigated sites located in a golf course (Monterey Country Club), and stormwater retention basins along a highway (El Dorado) and neighborhoods (Sonrisa, Harland) were initially surveyed for the following fire ant biological control agents: the microsporidian fire ant pathogen Kneallhazia solenopsae, Solenopsis invicta virus 3 (SINV-3), and fire ant decapitating phorid flies. Surveys were conducted in March 2014, and an additional site, irrigated turf between a street and parking lot (La Quinta Medical Center), was surveyed in October 2014. Surveys were conducted by collecting worker ant samples from 8-25 nests per site and using PCR to detect the pathogens or flies (Valles et al. 2010). In addition, the social form (i.e. monogyny or polygyny) was determined by Gp-9 analysis (Valles and Porter 2003). Three sites (Monterey, El Dorado, Sonrisa) were predominately polygyne, while Harland and La Quinta samples were 92 and 100% monogyne. The microsporidian K. solenopsae and the SINV-3 virus were each detected at three sites. There was no evidence of phorid flies in any of the samples (Table 1). Of the five sites, the Harland site was excluded from releases because most fire ant nests were located partially under a sidewalk which would hinder inoculations. K. solenopsae was released on May 8, 2014 at Monterey by introducing live, infected brood into 16 colonies following the procedures described in Oi and Valles (2009). While this site had a low initial K. solenopsae prevalence of 7.1% prior to inoculations, infections increased to 100, 70, and 71% at 12, 25, and 76 weeks after inoculation, respectively. Infections were detected from pupae samples several months after inoculations which indicated the pathogen had established because infections were occurring in progeny that were not from the pupae contained

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in the inocula. It should be noted that infections persisted despite irrigation being reduced at the Monterey site when landscapers tried to establish drought tolerant grass. In addition, K. solenopsae infection was found in 75% (6/8) of nests sampled at the La Quinta Medical Center, a new site over 7 miles away that was not inoculated. This suggested that K. solenopsae was well established at various locations in the Coachella Valley. In addition, CVMVCD had earlier records of low levels of K. solenopsae at other sites. For the fire ant decapitating phorid fly releases, a total of 64 g of fire ants (~64,000) were collected from 13 colonies at the Monterey site and 28 g of fire ants collected from 8 colonies at the Sonrisa site on May 8-9, 2014. These ants were exposed to parasitism attacks from large numbers of the little decapitating fly Pseudacteon curvatus and the larger decapitating fly Pseudacteon obtusus in Gainesville, Florida on May 12-14. Attacks were conducted in the field and at the USDA/APHIS rearing facilities at the Florida Division of Plant Industries. Parasitized ants were shipped back to California on May 13 and 14 for field release by CVMVCD staff into their mother colonies on May 15 and 16, 2014. Releases also were made at the La Quinta Medical Center site on Nov. 6-7, 2014. At the Monterey site, two male and one female P. obtusus were collected in October 2014 and Jan 2015, respectively. In May and October 2015, P. obtusus males and females and P. curvatus females were collected on sticky traps (Puckett et al. 2007). A majority of flies were P. obtusus males (65), followed by 9 female P. obtusus and 8 female P. curvatus. In May 2016 only 4 male P. obtusus were trapped, however that year had a severe drought. Collections increased dramatically in May 2017 consisting of 14 P. obtusus (3♀, 11♂) and 33 P. curvatus (30♀, 3♂). Flies trapped in 2017 were as far as 1/8 of a mile away from the release location. Flies were never collected at the Sonrisa or La Quinta sites after four samplings spanning 2 and 3 years, respectively. SINV-3 was released at the El Dorado site on June 11, 2014 and at the La Quinta Medical Center site on Jan. 26 and Oct. 22, 2015 using methods adapted from Valles and Oi (2014). SINV-3 was not detected in surveys of each of these sites prior to inoculation. Inocula consisted of dead, SINV-3 infected fire ants homogenized in a sucrose solution. The SINV-3 slurry was applied as a 50 ml drench on individual fire ant nests or applied as a bait by inserting a vial containing 12 ml of the slurry into a nest. Samples of foraging worker ants collected along transects in the vicinity of the treated nests at El Dorado and worker ants collected directly from inoculated and non-inoculated nests at La Quinta revealed that the virus had established at both sites. SINV-3 prevalence at El Dorado fluctuated between 9 and 48% from July 2014 to May 2016. At the La Quinta site, SINV-3 prevalence ranged from 18 to 68% from Feb. 2015 to May 2016. Infected samples were obtained beyond the vicinity of the inoculations, indicating SINV-3 was spreading. Evidence of the biocontrol agents impacting fire ant populations was not apparent based on the number and size of fire ant nests still present at sites where they established. However, that was expected due to the limited time of biocontrol establishment and their subtle impacts documented in the southern U.S. (Cook 2002, Oi and Williams 2002, Fuxa et al. 2005, Morrison and Porter 2005).

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In summary, releases of fire ant biological control agents were made in several irrigated urban landscapes in the Coachella Valley of California. The microsporidian pathogen, Kneallhazia solenopsae, Solenopsis invicta virus-3, and two species of the fire ant decapitating phorid flies, Pseudacteon obtusus and P. curvatus, were recovered for at least 3 years. All three types of agents increased in prevalence or spread from the original release locations, and have survived the extreme heat of the desert climate. Table 1. Fire ant social form and biocontrol prevalence at potential release sites sampled in March 2014. The Harland site was not used for releases.

Sonrisa El Dorado Monterey Harland La Quinta1 # nest samples 15 13 14 25 8 % monogyne 0.0 15.4 0.0 92.0 100

% SINV-3 53.3 0.0 28.6 8.0 0.0 % K. solenopsae 33.3 0.0 7.1 0.0 75.0

% phorid 0.0 0.0 0.0 0.0 --- 1Surveyed Oct. 2014. References Cited: Cook, T. J. 2002. Studies of naturally occurring Thelohania solenopsae (Microsporida:

Thelohaniidae) infection in red imported fire ants, Solenopsis invicta (Hymenoptera: Formicidae). Environmental Entomology 31(6): 1091-1096.

Fuxa, J. R., Y. Y. Sokolova, M. L. Milks, A. R. Richter, D. F. Williams, and D. H. Oi. 2005. Prevalence, spread, and effects of the microsporidium Thelohania solenopsae released into populations with different social forms of the red imported fire ant (Hymenoptera: Formicidae). Environmental Entomology 34(5): 1139-1149.

Morrison, L. W., and S. D. Porter. 2005. Testing for population-level impacts of introduced Pseudacteon tricuspis flies, phorid parasitoids of Solenopsis invicta fire ants. Biological Control 33(1): 9-19.

Oi, D. H., and D. F. Williams. 2002. Impact of Thelohania solenopsae (Microsporidia: Thelohaniidae) on polygyne colonies of red imported fire ants (Hymenoptera: Formicidae). Journal of Economic Entomology 95(3): 558-562.

Oi, D. H., and S. M. Valles. 2009. Fire ant control with entomopathogens in the USA, pp. 237-257. In A. E. Hajek, T. R. Glare and M. O'Callaghan (eds.), Use of microbes for control and eradication of invasive arthropods, vol. 6. Springer Science+Business Media B.V.

Puckett, R. T., A. Calixto, C. L. Barr, and M. Harris. 2007. Sticky traps for monitoring Pseudacteon parasitoids of Solenopsis fire ants. Environmental Entomology 36(3): 584-588.

Valles, S. M., and S. D. Porter. 2003. Identification of polygyne and monogyne fire ant colonies (Solenopsis invicta) by multiplex PCR of Gp-9 alleles. Insectes Sociaux 50(2): 199-200.

Valles, S. M., and D. H. Oi. 2014. Successful transmission of Solenopsis Invicta Virus 3 to field colonies of Solenopsis invicta (Hymenoptera: Formicidae). Florida Entomologist 97(3): 1244-1246.

Valles, S. M., D. H. Oi, and S. D. Porter. 2010. Seasonal variation and the co-occurrence of four pathogens and a group of parasites among monogyne and polygyne fire ant colonies. Biological Control 54(3): 342-348.

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APHIS Imported Fire Ant – Phorid Fly Rearing and Release Project 2001-2017

Anne-Marie Callcott (USDA-APHIS-PPQ)

and many, many cooperators History of the Project:

• In a late 1990s USDA-APHIS survey, 7 southern states ranked IFA as a top priority target organism for biological control

• Fire ant populations in native South America are 1/5 the density that they are in the U.S. • Phorid flies from South America (homeland of IFA) were the most promising biocontrol agent for

IFA at that time o Relatively specific to IFA o Active throughout the year o As many as 15-20 species or biotypes that will impact IFA

• Anticipated the flies being an important tool in the management of IFA – not a standalone control tool

o Allow native ants to compete with IFA for food and territory o Allow homeowners, etc. to make fewer insecticide treatments o Suppress IFA to acceptable tolerance level

Killing some IFA worker ants Impacting IFA foraging activity through attacks Being possible vectors/carriers of pathogens

In 1994, two independent projects were initiated in Brazil:

1. USDA-ARS/Porter (Gainesville, FL) o focused on phorid fly life history studies and rearing methods o in collaboration with Harold Fowler at Sao Paulo State University in Rio Claro

and Luiz Alexandre Nogueira de Sa at the Embrapa research center near Jaguariuna, SP, Brazil

o first permits to release in U.S. in 1997 2. University of Texas/Gilbert (Austin, TX) o focused on ecological studies of fire ants and phorid flies o in cooperation with Woodruff Benson at the University of Campinas, Brazil o and later with Patricia Folgarait (Universidad Nacional de Quilmes, Buenos Aires,

Argentina) o first permits to release in U.S. in 1995

With the success of the ARS and UT releases, in 2001, USDA-APHIS initiated a cooperative program to rear and release decapitating flies as IFA biocontrol agents in all infested states

• USDA-APHIS-PPQ (Biloxi Station) o coordinated activities

• USDA-ARS-CMAVE, Gainesville, FL o Explored in South America; imported; developed rearing methods; conducted

preliminary releases; transferred rearing technology to FL DPI • Florida Dept. of Ag.-DPI reared and shipped flies

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o Pseudacteon tricuspis: prefers red IFA and attacks workers on/around mound o Pseudacteon curvatus: prefers smaller red IFA (polygyne) o Pseudacteon obtusus: prefers very large workers o Pseudacteon culltelatus: prefers small workers and attacks on foraging trails

• Cooperators o State cooperators, such as universities, state department of agriculture, etc.

handled releases and monitoring APHIS Releasers/Cooperators: Primary contacts – lots of other participated:

• Alabama – Fudd Graham (Auburn) and extension agents and students • Arkansas – Kelly Loftin (Univ. Ark) and extension agents and students • California – Kris Godfrey (CDFA); PPQ staff • Florida – a lot of FL extension agents • Georgia – Wayne Gardner (Univ. Ga) • Louisiana – Seth Johnson (LSU) and students • Mississippi – MS-DPI; Anne-Marie Callcott and PPQ staff • North Carolina – Kathleen Kidd (NCDA) • Oklahoma – Don Molnar (OK Ext); Russell Wright (OSU); Jeanetta Cooper (OK-DPI) • Puerto Rico – Nilda Perez (PR-DPI) • South Carolina – Tim Davis (Clemson ext); Steve Compton (SC-DPI) • Tennessee – Jason Oliver (TSU) • Texas – Bart Drees/Robert Puckett (TAMU) and a lot of TX ext agents • Virginia – Gina Goodwyn/Larry Nichols (VA-DPI)

• Lots of ARS releases throughout the U.S. and lots of UT releases throughout Texas –

both with multiple species of releases Rearing of Phorid Flies: Rearing of these flies is extremely labor intensive, requiring 1-1.5 person(s) to maintain every 2 attack boxes. These flies cannot be reared on a special diet or medium but require live fire ants to complete their life cycle. An excellent pictorial and text description of the rearing technique is available online from the FL DPI at: http://www.freshfromflorida.com/Divisions-Offices/Plant-Industry/Science/Biological-Control/Imported-Fire-Ant-Biological-Control Release of Phorid Flies: Release techniques for the flies evolved over the years. P. tricuspis was released by shipping pupae to the field and having the cooperator rear the flies out to adults (within a few days of shipping) in rearing cages. The adults would then be collected, hand carried to the field and released over disturbed mounds. To release the other species, worker IFA were field collected from the release site, shipped to the FL-DPI facility, exposed to flies, and then shipped back to the field to be returned back to their “home” mound. In later years, ants were separated by size and exposed to different flies, therefore many site releases were composed on 2 or even 3 species of flies. Releases by year are shown in Figure 1.

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Figure 1. Releases per year of phorid flies by species.

Monitoring of flies: Monitoring was originally done at release sites, then moving out symmetrically to determine spread. However, as more and more releases were done by multiple groups, it became more practical to survey at the county level for presence or absence. Distribution of Flies: Through APHIS releases, along with other federal and university releases, P. tricuspis is well established in the southern areas of the IFA regulated area and is estimated to cover ca. 70% of the IFA regulated area, although populations have declined in recent years. To date, P. tricuspis is not known to be established in CA, NM, TN or VA. The second species, P. curvatus, is well established in most southern IFA regulated states and PR, and is estimated to cover more than 90% of the regulated area. Establishment of P. obtusus has occurred in small areas of GA and MS, and fairly large areas in FL and TX, and in 2014 it was established in CA. P. obtusus is estimated to cover about 25% of the regulated area. P. cultellatus has been confirmed in FL with limited expansion at this time. In 2013, the Imported Fire Ant community of practice within the e-Xtension website began compiling information on phorid fly distribution from all sources, as well as estimates of spread based on data from Dr. Sanford Porter and others. They have continued adding data as federal and state cooperators, universities and others publish data or provide unpublished survey data, in an effort to keep fairly current maps of distribution of six species of phorid flies. These interactive maps are available at: http://articles.extension.org/pages/30546/natural-enemies-of-fire-ants. This site also provides information on some other potential biological control agents for IFA.

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Conclusion: The APHIS-funded phorid fly rearing and release project will be doing its last releases in July 2017. We would like to thank the following for their continued support of this successful project over the last 16 years. Special Thank You….. FL-DPI: George Schneider, Debbie Roberts, Amy Croft, Amy Bass and Catherine White ARS: Sanford Porter University of Texas: Larry Gilbert, Rob Plowes And to all the cooperators

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The Tawny Crazy Ant (Nylanderia fulva) at the Port of Savannah, Georgia

Ben Gochnour & Dr. Dan Suiter Department of Entomology, University of Georgia, Griffin, Georgia.

Invasive species are an economic and ecological threat. Port cities play a significant role concerning the introduction of potentially invasive species into and out of North America, making them important research sites. Recently, the Tawny Crazy ant (Nylanderia fulva) was found on the Port of Savannah, Georgia. Though the species appeared to be restricted to several wooded areas on the port’s property, the potential for expansion was great given the species’ documented ability to spread rapidly. Consequently, the objective of this study was to determine the rate of expansion of the Tawny Crazy ant on the Port of Savannah property and to evaluate the impacts of the species on the ant communities in the area. Intensive sampling of the ant communities in the wooded areas (pitfalls, leaf litter, baits, tree trunks, and logs) as well as along roadsides (baits and quadrats) was carried out from July 2015 to July 2016. A total of 48 species across 19 genera were found on the port. Of the 48 species, 14 were exotic across 10 genera. In the wooded areas on the port the Tawny Crazy ant reduced ant species richness, homogenizing the ant community. However, its effect appeared to be non-random. Larger, ground foraging species were the most susceptible to extirpation by the Tawny Crazy ant, while very small, cavity dwelling species and arboreal nesting species showed the most resistance in invaded areas. The Red Imported Fire ant (Solenopsis invicta) was readily eliminated from both wooded areas and roadsides following invasion by the Tawny Crazy ant. Baiting and quadrat sampling along the roadsides revealed the competitive exclusion of the Red Imported Fire ant by the Tawny Crazy ant through resource domination. Additionally, it was determined that the Tawny Crazy ant was moving along the highly disturbed roadside habitats faster than through the wooded areas on the port. This difference in movement led to the ant first surrounding the wooded areas, then moving into them and affecting the ant communities within. The results of this study may implicate small scale disturbance corridors such as dirt roads, trails, and fire breaks in the facilitation of the invasive Tawny Crazy ant’s movement into undisturbed habitats.

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Alleged Injury to New Born Calves in Texas Counties Attributed to Presence of Nylanderia fulva (tawny crazy ant)

Paul R. Nester1 and Robert T. Puckett2,

1Texas A&M AgriLife Extension Service, Houston, TX 2Texas A&M AgriLife Extension Service, College Station, TX

In late summer of 2015 and 2016, injury to new born calves (Figures 1, 2, and 3) was observed in both Brazoria and Wharton Counties in Texas and attributed to the presence of the tawny crazy ant (Nylanderia fulva). Possible strategies to limit the impact of the tawny crazy ant on new born calves are outlined below. Strategies for protecting newborn calves

1) In a TCA infested area it is best to plan calving before May and after October 2) If this is not possible it is suggested to prepare a confined calving pasture to minimize the

impact of the TCA 3) Depending on the number of animals to be confined, set up the confined pasture area in

2-3 separate lots so animals can be rotated from lot to lot to reduce foraging pressure on the total confined pasture area

4) Choose and prepare an area for a confined calving pasture so broadcast insecticide treatments can be applied to the grounds to reduce ant activity and allow for a stress free calving

Strategies for preparation of confined pasture areas

1) Do not place confined pasture area against a wooded area of land. This places the confined pasture away from areas of TCA refuge and over-hanging limbs

2) Leave a 100 ft. buffer from edge of wooded area to fence of confined pasture to allow easy treatment of ground around the confined area

3) Remove any debris, i.e., limbs, wood, any unnecessary materials, from within and around the confined pasture area.

Management strategies for tawny crazy ant in confined pasture areas

1) Select insecticides with approved active ingredients for use around and on livestock. 2) Begin broadcast insecticide treatments with lowest labeled rate in and around the

confined calving pasture one month before calving. 3) Continue monitoring weekly to make sure insecticide treatments are effective. 4) Once ant populations start to increase, move targeted cattle to one of the treated confined

pasture lots. 5) Rotate stock from one split pasture lot to the other every two weeks. This will reduce

foraging pressure within lots and also allow treatments to lots without livestock being present.

6) Treat lots every 2 weeks and continue until young are up and walking and able to move freely in pastures.

7) Application rates and intervals may need to be adjusted depending on weekly monitoring observations.

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8) Stock density will decrease as calves are more mobile and can be moved to a permanent pasture

9) As stock density decreases and the young are moved to other pastures, foraging pressure will be reduced within the confined pasture area.

10) Treatment to young calves with approved insecticide may be necessary. 11) Contact veterinarian for suggestions on treatment of symptoms if necessary.

Monitoring for TCA presence

1) Begin monitoring tawny crazy ant populations in April, in and around the confined calving pasture

2) A ¼ inch thick slice of a large diameter or “jumbo” hot dog is used as a monitoring food lure.

3) Usually a slice from a jumbo sized hot dog gives enough area to observe the presence of TCA

4) Place lures every 10 – 15 steps around the border and within the confined pasture area. 5) Mark the lures with an electrician’s flag to aid in equal placement of lures and their

recovery. 6) Let stand for 20 minutes then observe for the presence of the TCA (Figures 4 and 5) 7) The denser the population of TCA, the quicker the lures will be recruited to. 8) If more than 50% of the food lures have TCA present within the 20 minute period, it is

time to treat. If any of the lures have TCA present, monitor every week.

Figure 1: Suspected TCA injury to new-borne calf. Ants present around the eye, Brazoria, TX, August 2015.

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Figure 2: Suspected TCA injury to new-borne calf, Brazoria, TX, August 2015

Figure 3: Suspected TCA injury to new-borne calf with ants present in corner of eye, Brazoria, TX, August 2015.

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Figure 4: Hot dog food lure with TCA present, good time to plan to treat

Figure 5: Excessive number of TCA on hot dog food lure

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Fire Ant Control Demonstration at Storybook Farm

Mallory Kelley¹ and L. C. ‘Fudd’ Graham² ¹Regional Home Grounds Horticulture Agent, Alabama Cooperative Extension System,

Autaugaville, AL 36003 ²Department of Entomology and Plant Pathology, Auburn University , Auburn, AL 36849

Abstract

Storybook Farm, located in Opelika, Alabama is a non-profit organization serving children ages two to young adulthood who face obstacles such as autism, cerebral palsy, cognitive delays, sensory integration issues and bereavement situations to name a few. Horseback riding provides physical benefits such as improvements to balance, motor skills, muscle strength and coordination. Due to the popularity of this venue and those it serves, public health concerns must be addressed. Fire ant management has been a challenge on this 25-acre site. In 2015 we were notified of the problem through a volunteer at the facility. Extension then reached out to Storybook to develop a strategy to address this public nuisance. After a site assessment, an application of fire ant bait was chosen as the control measure of choice for the farm. Pre-treatment mound counts were taken on four ¼ acre plots and Extinguish Plus® bait was applied in October of 2015. Data were collected four, eight and twelve weeks post-application. By week twelve, fire ant mound numbers were reduced by 80 percent. A second bait application was made in October 2016. November data indicated we had achieved 97 percent control compared to original numbers in 2015. Storybook Farms director, Dena Little has been thrilled with the extremely effective control using the bait application method and especially the safe environment it has provided at the facility.

Introduction

We contacted Storybook Farm at the request of the farm’s veterinarian Dr. Debra Taylor, ambulatory clinician at Auburn University. One of the horses, Dr. Dolittle, had rolled in a bed of fire ants resulting in a massive amount of hives covering a large portion of his body. This wasn't the first time they had encountered a fire ant infestation and subsequent trauma for both animals and staff members. Fire ants had literally taken over the paddocks and programming areas. This posed a danger, not only to the horses, but also the volunteers and clients being served. They had tried various measures for control in the past with no success. We discussed products, application methods and timing of control. We then set up a demonstration to begin control of the fire ant population at Storybook Farms.

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Methods

Four ¼-acre circular plots were selected for data collection to determine the effectiveness of our demonstration treatments, one by the riding paddock and three in pastures. Broadcast applications of Extinguish Plus®, donated by Central Life Sciences, were made at a rate of 1.5 lbs./A using a Herd® GT-77 Bait Spreader on 10/6/15. Data were collected pre-treatment and four, eight and twelve weeks post-treatment. A second bait application was 10/4/16 and data were collected again six weeks post-treatment. Mound data were collected by minimally disturbing and visually surveying mounds in the four ¼ acre plots.

Results

On 10/6/15, Storybook had an excessive number of fire ants on the property with a mean of 127 mounds per acre (Fig 1). The average number of mounds per acre in Alabama ranges from 40-80. Eight weeks post-treatment, only 46 mounds per acre were present. By week twelve, fire ant mound numbers were reduced by 80 percent. One year later, on 10/4/16, 16 mounds per acre were found in the plots. A second bait application was made after sampling 10/4/16. November data indicated we had achieved 97 percent control compared to original numbers in 2015.

44

Impacts

Storybook Farms is a place where children challenged with a variety of physical, emotional, social and mental situations experience the exhilaration of riding. Fire ants cause painful stings and create a hazardous situation for children, volunteers and animals at Storybook. Founder and executive director, Dena Little, had this to say about the demonstration, “Our treatments had produced little to no result and had not abated their resurgence at all. In sharp contrast to our endeavors, these treatments dramatically reduced Storybook's fire ant population. Where I once saw dozens of mounds, I only saw one or two. Because of their help, we have had no additional incidents with fire ants. Their efforts truly changed the landscape at Storybook Farm and we remain grateful for their expertise.”

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Figure 1. Mean number of mounds per acre at Storybook Farms. Percent control is indicated by the blue line.

45

Evaluation of Imported Fire Ant Quarantine Treatments in Commercial Grass Sod: Arkansas 2016

Kelly M. Loftin1, John D. Hopkins2, and Anne-Marie Callcott3

1 Univ. of Arkansas System, Div. of Agric., CES, Cralley-Warren Research Center 2601 North Young Ave., Fayetteville AR 72704, 2University of Arkansas System, Division of Agriculture, CES, 2301 S. University Ave., Little Rock, AR 72204, 3USDA, APHIS, PPQ, CPHST-Gulfport Laboratory, Imported Fire Ant Section, Gulfport, MS 39501 Abstract In 2016, studies were conducted in SW Arkansas to evaluate potential quarantine treatments against imported fire ants in commercial sod. Applications of Taurus Trio with and without a prior Advion bait application were compared to a tank mix of carbaryl/bifenthrin with a prior Advion bait treatment and an untreated control. All insecticide treatments significantly reduced the number IFA colonies in treated plots. The Advion® plus Taurus® Trio G and the Taurus® Trio G treatments provided six and seven weeks of quarantine-level control, respectively. Data suggests that Advion® fire ant bait reduced the exposure period by one week. Introduction Imported fire ants (IFA) originated from South America and were accidentally introduced into the United States in the early to mid-1900s. IFAs are now widespread across the Southeastern United States. Movements of this pest are regulated through a system of Federal and State quarantines. Products regulated by the IFA quarantine include but are not limited to hay, nursery plants and other landscape materials including grass sod. When treating sod in compliance with Federal and State quarantine regulations, sod producer’s options are limited (USDA-APHIS 2006). One option was treatment using the active ingredient chlorpyrifos at a rate of eight pounds of active ingredient per acre. Currently, no chlorpyrifos products are registered for IFA in sod at that required rate. Another option is to use two separate applications of fipronil at 0.0125 pounds per acre about one week apart. Fipronil can be too expensive to apply and the longer required exposure period (30 days) can be a logistical problem for sod producers. One recently approved quarantine option is two applications of 0.2 lb ai/acre bifenthrin, one week apart, for a total of 0.4 lb ai/acre. This option is less costly than fipronil and has a slightly shorter exposure period (28 days) than fipronil. Because of limited or costly options available to sod producers, a field study was conducted to evaluate the efficacy of other insecticides for use in the IFA quarantine. Using fire ant bait as the first application, followed by 0.2 lb.ai/acre of bifenthrin or the combination of 0.17 lb.ai/acre bifenthrin and 0.35 lb.ai/acre clothianidin has shown much promise as a quarantine treatment. The current study is to evaluate another commercial combination product – Taurus® Trio G (fipronil, bifenthrin and lambda cyhalothrin) with and without a prior (7 days) fire ant bait application with Advion. In addition, a tank mix of bifenthrin and carbaryl with a prior Advion fire ant bait application was evaluated.

46

Materials and Methods The study was conducted on an irrigated sod farm in Fulton, AR (Hempstead Co.) beginning in late July 2016 and ending in October 2016. Plots were square, measuring ½ acre in area, and treatments (three treatments and an untreated control) were arranged in a Randomized Complete Block Design (RCBD) with three replications. Plots used in the study had a range of 16-40 active fire ant mounds per acre when the study began. An active fire ant mound is defined as a mound with 25 or more ants in the colony which is the USDA standard for classifying active mounds. In plots receiving two treatments (bait/application timing #1 followed by the carbaryl/bifenthrin combination or Taurus® Trio G/application timing #2), application timing #1 and application timing #2 were separated by seven days. In plots receiving only the Taurus® Trio G product, treatments were applied at application timing #2. Spray applications were made using a towed boom sprayer applying @ 20 gal/A (15 ft. boom with ten 8003FF nozzles on an 18" spacing at 20 psi and 5.2 MPH). Granular bait applications were made using a Herd fire ant spreader attached to a Kawasaki Mule ATV and calibrated to apply 1.5 pounds per acre. Granular fipronil/bifenthrin/lambda cyhalothrin (Taurus® Trio G) applications were made using a tow-type granular applicator (Agri-Fab) towed by a Yamaha ATV and calibrated to apply 87 pounds per acre. Treatment numbers, insecticide rates and the total amount of active ingredients applied per acre are provided in Table 1. Table 1. Insecticide applications, rates and total amount of active ingredients.

Treatment Number

Insecticide Application

Total active ingredients/acre

1 None – Untreated Control None 2 1 application Advion® 0.045% bait (1.5 lb./A) and

1 application of Taurus® Trio G at 87 lb/acre applied 7 days after bait

0.000675 lb ai/A indoxacarb 0.0124 lb ai/A fipronil 0.2 lb ai/A bifenthrin 0.051 lb ai/A lambda cyhalothrin

3 1 application of Taurus® Trio G at 87 lb/acre 0.0124 lb ai/A fipronil 0.2 lb ai/A bifenthrin 0.051 lb ai/A lambda cyhalothrin

4 1 application Advion® 0.045% bait (1.5 lb./A) and 1 application of tank mix Onyx Pro at 13.9 oz./A and Sevin SL at 128 fl oz/A applied 7 days after bait

0.000675 lb ai/A indoxacarb 0.2 lb ai/A bifenthrin 4.0 lb ai/A carbaryl

The number of active mounds per plot was determined by counting the mounds in a circle at the of the center plot. This circle had a diameter of 58.9 ft which corresponds to a circle with an area of 0.25 acre. The mounds were counted by anchoring one end of a 58.9 ft. rope at the center of the plot and moving the free end along the circumference of the circle. Each mound encountered anywhere along the length of the rope was disturbed by probing with a small rod and estimating the number of imported fire ants exiting the mound within 20 seconds (Jones et al 1998). The number of active mounds in each plot was determined before any treatments were applied and then at seven days after the last application (DALA) then weekly up to 28 DALA, at which time evaluations were made every 14 days, through early October.

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All data were analyzed using Gylling’s Agriculture Research Manager Software (ARM 7.0.3. 2003). An analysis of variance was performed and Least Significant Difference (p=0.05) was used to separate means only when AOV Treatment P(F) was significant at the 5% level (ARM 2003). Results and Discussion The data are summarized on Table 2 and Figure 1. Before applying treatments, there were no significant differences in the number of active mounds in any of the plots used in the study. Throughout the remainder of the study, all insecticide treated plots had significantly (p<0.05) fewer active IFA colonies compared to the untreated control. At eight through 42 DALA, the Advion® bait plus Taurus® Trio G treated plots had zero active mounds per acre. The Taurus® Trio G-only treated plots had no active mounds at 14 through 56 DALA. For the Advion® fire ant bait followed by the bifenthrin/carbaryl tank mix treated plots, no fire ant colonies were found at the eight through 21 DALA evaluations. Fire ant colonies returned to the plots in the Advion® plus Taurus® Trio G and Taurus® Trio G-only plots at 56 and 70 DALA, respectively. Although the Advion® plus bifenthrin/carbaryl treated plots had significantly fewer active colonies than the untreated control, eight through 21 DALA were the only evaluation periods with no active colonies. Untreated controls maintained reasonable fire ant activity all summer, probably due to routine irrigation of the test area. All insecticide treatments significantly reduced the number IFA colonies in treated plots throughout the study duration and are suitable for control in home lawns, parks or recreational areas. The Advion® plus Taurus® Trio G and the Taurus® Trio G treatments provided six and seven weeks of quarantine-level control, respectively (quarantine level control is defined as no active colonies). Data indicated that the addition of Advion® fire ant bait reduced the exposure period by one week. This study indicated that the Taurus® Trio G treatment demonstrates the level of control necessary for shorter-term (six weeks) fire ant quarantine treatment of grass sod. References ARM 7.0.3. 2003. Gylling Data Management, Inc. Brookings, SD. Jones, D., L. Thompson and K. Davis. 1998. Measuring Insecticide Efficacy: Counting Fire Ant Mounds vs. Bait Station Sampling. In Proceedings of the 1998 Imported Fire Ant Conference. Hot Springs, Arkansas. pp. 70-78. USDA-APHIS. 2015. Imported Fire Ant: Quarantine Treatments for Nursery Stock, Grass Sod and Regulated Materials. APHIS 81-25-001.

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Table 2. Average Number of Active Mounds/0.25 acres for each treatment.

Treatment Average number of active colonies per 0.25 acre Pre

Treat 8

DALA 14

DALA 21

DALA 28

DALA 42

DALA 56

DALA 70

DALA 84

DALA UTC 7.3a 6.3a 5.7a 8.0a 7.7a 5.0a 5.7a 9.7a 10.3a Advion® 0.045% bait (1.5 lb./A) followed by Taurus® Trio G at 87 lb/acre applied 7 days after bait

7.3a 0.0b 0.0b 0.0b 0.0b 0.0b 0.3b 0.3b 0.3b

Taurus® Trio G at 87 lb/acre

6.0a 0.3b 0.0b 0.0b 0.0b 0.0b 0.0b 0.3b 0.0b

Advion® 0.045% bait (1.5 lb./A) followed by tank mix of Onyx Pro at 13.9 oz./A and Sevin SL at 128 fl oz/A applied 7 days after bait

5.7a 0.0b 0.0b 0.0b 0.7b 0.3b 1.7b 1.0b 1.3b

Means followed by same letter do not significantly differ (P=.05, protected LSD) Figure 1. Average Number of Active Mounds/0.25 Acres for each treatment.

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An Estimation of Temperature Limits for the Tawny Crazy Ant, Nylanderia fulva (Hymenoptera: Formicidae)

David C. Cross and Michael A. Caprio

Biochemistry, Molecular Biology, Entomology and Plant Pathology, Mississippi State University Introduction Tawny crazy ant, Nylanderia fulva, has a native range in a south-central region of South America that has been identified as a hotbed for producing many invasive ant species. It has spread to the Galapagos, several parts of the Caribbean islands and pockets of Mexico and Central America. It likely came to the U.S. mid-20th century, arriving in Florida. Currently this species is in 30 counties in TX, 3 in MS, 2 in AL, 6 in GA, 28 in FL and 25 parishes in LA. Making predictions for the limits of introduced ranges for invasive ants has been shown to be prone to error, as evidenced by early predictions for the imported fire ant in the U.S. We hope to add timely information useful for both gauging physical limitations of tawny crazy ant (TCA) as well as prompting more investigation in its behavioral plasticity that may be contributing to this nuisance ant’s expansion in its non-native range.

Fig. 1. U.S. Counties with records of N. fulva Materials and Methods Worker ants were collected from TCA colonies in Jackson Co., MS, during different seasons in 2014-2015. They were maintained in small, covered containers with a choice of 10% honey water, tap water and an artificial diet (Dussutour and Simpson, 2008, modified). The holding container walls were coated with Insect-a-Slip (BioQuip). The ants were kept at 80-95% relative humidity and temperatures from 16-28 °C. Chilling injury protocol: Ants were placed in 4 ml glass vials with moistened paper toweling in the cap. Vials were held in an aluminum block (10 x 11.8 x 5 cm) within an insulated 13x13x5

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cm box containing sensors from a Pico TC-08 data logger and a Fluke 54 II thermometer. The box was placed in a frost-free freezer at the appropriate level for each temperature and at differing durations. Ants were scored at 30 min. and again after 24 hrs. after returning the ants to a 20-25 °C ambient temperature. Those that would walk and move normally after light prodding were scored as “live”. All others were scored “dead”. Heating injury: MacDonald (2012) reported that TCA foraging ceases if ground temperatures reach 37.3 °C. We placed test boxes in an environmental chamber for differing durations at 40 °C and recorded their ability to recuperate post treatment. Mimicking environmental buffering: After observing TCA possibly sheltering from winter cold in partially rotted pine logs in coastal MS, we measured within log temperature versus that of the ambient air in 3 pine logs or log sections at sites in northeast MS. We drilled holes into the interior and placed a T–thermocouple within and in air at an adjacent shaded spot. We logged temperatures during cold weather events using a Fluke 54 II thermometer.

Fig. 2. Chilling injury recording set-up.

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Fig. 3. Typical chilling injury temperature profile. X-axis is time (hours).

Fig. 4. Chilling injury survivorship. Probit analysis for survival probability of TCA workers using exposure of 17.3 hours within 1 °C of final low temperature, (95% Confidence Limit).

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Fig. 5. Heating injury survivorship. Probit analysis for survivorship of TCA workers at differing durations for exposure at a temperature constant of 40.3 °C, (95% Confidence Limit).

Fig. 6. Cold aversion via habitat choice? Interior temperatures of rotting logs recorded during cold weather events.

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Fig. 7. Air versus within log temperature. 7a. Log: 34cm diameter and 11cm depth for sensor

7b. Log: 46cm diameter and 12cm depth for sensor

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Results and Discussion Chilling injury: 1173 worker ants were tested in 42 trials. At a test duration of 17.3 hours, the data suggest an LT50 (temperature with 50% mortality) of -6.6 °C (Fig. 4). Estimations of the metabolic cost for an insect’s recovery from chill-coma have been shown to be a significant drain of resources. Given real-time weather patterns, cycling in and out of this critical range is likely limiting for colony growth and reproductive success. Heating injury: For the 389 workers tested in 5 trials, we found the “LD50 ” for the 40.3 °C test temperature to be approximately 4.6 hours. Microhabitat choice shift: Nylanderia fulva has been shown to favor microhabitats with moderate to high protection from temperature and moisture extremes, nesting in areas such as layered refuse with soil contact, cavities in tree roots, under bark or even in termite galleries within rotting logs. Having found TCA moving in abandoned termite tunnels at least 8-10 cm within a rotting pine snag during a winter event in 2011, we wished to simulate the habitat for cooler temperatures. Depending on depth within the wood, interior temperature change lags in duration for both cooling and warming phases. The greater buffering from greater depth may protect ants from death from freezing, but also might prevent movement and foraging within a sufficient time during the warming phase. The question that arises is what behavioral responses does the TCA have to ameliorate environmental conditions and to what degree are they effective? Given the likelihood they can moderate these extremes, care must be taken when making predictions on limits to TCA range expansion. One might consider peculiarities of microhabitat factors, such as

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considering a source of metabolic heat from wood decay or other organisms and their effect on temperature or other environmental conditions. Areas for additional study:

• Repeated chill events more closely relating to real weather patterns • Survivorship at multiple moisture levels • Extensive evaluation of an upper thermal limit • Quantify the ability of a colony to produce sufficient numbers of sexual caste members to

expand the colony in these colder (or hotter or drier) conditions • Devise experiments to gauge behavioral response within protected environments against

physical extremes Literature Cited Berman, D.I., A.V. Alfimov, Z.A. Zhigulskaya and A.N. Leirikh. 2010. Overwintering and

cold-hardiness of ants in the northeast of Asia. Pensoft Publishers, Sofia, Bulgaria. Dussutour, A., and S.J. Simpson. 2008. Description of a simple synthetic diet for studying

nutritional responses in ants. Insectes Sociaux 55: 329-333. Gotzek, D., S.G. Brady, R.J. Kallal, and J.S. LaPolla. 2012. The importance of using multiple

approaches for identifying emerging invasive species: the case of the rasberry crazy ant in the United States. PloS One, 7(9):e45314.

MacDonald, D.L. 2012. Investigation of an invasive ant species: Nylanderia fulva colony extraction, management, diet preference, fecundity, and mechanical vector potential (Ph.D. Dissertation). Department of Entomology, Texas A&M University, College Station. p. xi + 150.

MacGown, J.A. Ants (Formicidae) of the Southeastern United States. 14 Jan. 2013. Mississippi State University. 25 Oct., 2013. <http://www.mississippientomologicalmuseum.org.msstate.edu/Researchtaxapages/Formicidaepages/genericpages/Nylanderia_fulva.htm>

Acknowledgements Trials and care of insects were carried out with the assistance of Rob Gilmore and Dillon Coulter. This research was supported in part by the National Institute of Food and Agriculture, U.S. Department of Agriculture, under Project No. MIS-012040 and the USDA-ARS Area-wide Management of Imported Fire Ant Project.

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Comparative Water Relations and Desiccation Tolerance of Five Invasive Ant Species

Olufemi Ajayi1, Arthur Appel1, Chen Li2, Rammohan Balusu1, and Henry Fadamiro1

1Auburn University, 301 Funchess Hall, Auburn. AL 36849-5413 2Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, China

The success of invasive insects can be linked to their impressive tolerance to environmental stress, but little is known about such responses in the invasive tawny crazy ant, Nylanderia fulva (Hymenoptera: Formicidae), a rapidly emerging economically important species in the southern United States. Nylanderia fulva is sympatric with the alien-invasive Solenopsis species (Hymenoptera: Formicidae) in their native South America. Determining the tolerance of N. fulva to environmental stress may provide insight into its invasive potential. Here, we examined the water relations and desiccation tolerance of N. fulva relative to that of the introduced S. invicta, S. richteri, S. invicta × S. richteri; and native S. geminata. We compared these variables among size-classes and species. Results show that cuticular water loss is significantly more important to overall water balance than respirometry water loss. This study provides insight on the adaptability and potential distribution of invasive ant species. The ecological significance and practical implications of the results are discussed.

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Relationship of Imported Fire Ant (Solenopsis spp.) Integument Coloration to Cuticular Hydrocarbon and Venom Alkaloid Indices

Manoj Pandey,1 Jason Oliver, 1 Karla Addesso, 1 Steven Valles, 2 and Anne-Marie Callcott3

1 Tennessee State University, College of Agriculture, Human, and Natural Sciences, 3500 John A. Merritt Boulevard, Nashville, TN.

2United States Department of Agriculture - Agricultural Research Service, Center for Medical, Agricultural and Veterinary Entomology (CMAVE), 1700 SW 23rd Drive, Gainesville, FL.

3United States Department of Agriculture – Animal, Plant Health Inspection Service – Plant Protection and Quarantine, Biloxi Station, 1815 Popps Ferry Road, Biloxi, MS.

This study investigated the relationship of imported fire ant (Solenopsis spp.) (IFA) integument coloration to cuticular hydrocarbon and venom alkaloid indices used in IFA species assessment in an effort to facilitate faster identification of black (Solenopsis richteri Forel) and red (Solenopsis invicta Buren) IFA species and their hybrid. The identification of the IFA species and hybrid is important to match with appropriate classical biocontrol agents and to determine their regional distribution to guide future biocontrol strategies. Worker ant samples were collected from 62 IFA infested Tennessee counties, cuticular hydrocarbons and venom alkaloids were analyzed using gas chromatography and mass spectrometry, samples grouped into colony cohorts based on hydrocarbon and alkaloid values, and integument color quantified using a Konica Minolta CM-2600d spectrophotometer. The L* (lightness), a* (green/red), b* (blue/yellow), C* (chroma), and h* (hue) values, as well as CIE L*a*b and CIE L*C*h differences from red IFA standards, were measured and then analyzed for differences among colony cohorts (International Commission on Illumination [CIE], Vienna, Austria). Significant color differences were found among alkaloid and hydrocarbon colony values, particularly between black and red IFA. Hybrid colonies with increasing cuticular hydrocarbon and venom alkaloid values going from black to red IFA did not follow an expected linear change in the different color attributes measured. The change in color space (L*a*b* or L*C*h*) from the red IFA standard did not yield distinct patterns that could be used to predict IFA species or hybrid. Individual color attributes (L*, a*, b*, C*, or h*) may be a better predictor of IFA species or hybrid, but there was some overlap in color values among hybrid colonies with different cuticular hydrocarbon and venom alkaloid values. Because of inconsistencies in color space values among black, red, and hybrid IFA cohorts, integument coloration may not be a reliable method for identification of IFA species and their hybrids. Thus, further studies are needed. We thank USDA-APHIS-PPQ for providing partial funding of this study (Cooperative Agreement Award 15-8130-0523-CA).

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2017 IFA and Other Invasive Ant Conference Attendees Olufemi Ajayi Auburn University 301 Funchess Hall, Auburn University Auburn, AL 36830 osa0001@auburn.edu Charles Barr Barr Research and Consulting 1508 Caudill College Station, TX 77840 clbarr@verizon.net Vicky Bertagnolli Clemson Extension 1555 Richland Avenue East, Suite 500 Aiken, SC 29801 vbertag@clemson.edu Awinash Bhatkar Texas Dept of Agriculture PO Box 12847 Austin, TX 78711 Rhonda Britton ACES 819 Cook Ave Huntsville, AL 35801 RCB0003@aces.edu Anne-Marie Callcott USDA, APHIS, PPQ 1815 Popps Ferry Rd Biloxi, MS 39532 anne-marie.a.callcott@aphis.usda.gov Charlie Cassidy Orange County Mosquito & Vector Control District 13001 Garden Grove Blvd Garden Grove, CA 92843 ccassidy@ocvcd.org

Jian Chen USDA-ARS 59 Lee Road Stoneville, MS 38776 jian.chen@ars.usda.gov Stephen Compton Clemson University 511 Westinghouse Road Pendleton, SC 29670 scompto@clemson.edu Sydney Crawley The Scotts Miracle-Gro Company 14111 Scottslawn Rd Marysville, OH 43041 sydney19@gmail.com David Cross Mississippi State University 100 Old Hwy 12 Mississippi State, MS 39762 dcc6@msstate.edu Everett Dale USDA, APHIS, PPQ 301 NW 6th Street; Suite 101 Oklahoma City, OK 73102 everett.dale@aphis.usda.gov Willie Datcher, Regional Extension Agent ACES 107 Harris Avenue Eutaw, AL 35462 datchwe@auburn.edu Bob Davis BASF 2605 Butler National Dr. Pflugerville, TX 78660 robert.davis@basf.com

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Timothy Davis UGA Cooperative Extension - Chatham Co. 124 Bull Street, Suite 120 Savannah, GA 31401 tdavis@uga.edu Kimberly Dickinson Central Garden & Pet 7625 Woodhaven Drive North Richland Hills, TX 76182 kdickinson@central.com Bobbye Dieckmann CVMVCD 43420 Trader Place Indio, CA 92201 cmoreno@cvmvcd.org Jessica Dieckmann 43420 Trader Place Indio, CA 92201 cmoreno@cvmvcd.org Henry Dorough Auburn University 130 North Court Street Talladega, AL 35160 dorouhd@auburn.edu Yuzhe Du USDA-ARS, BCPRU 59 Lee Road Stoneville, MS 38776 yuzhe.du@ars.usda.gov Lucy Edwards Alabama Cooperative Extension System 202 South Hwy. 123 Ste. D Ozark, AL 36360 lee0003@auburn.edu Kathy Flanders Auburn University 201 Extension Hall Auburn University, AL 36849 flandkl@auburn.edu

Ben Gochnour University of Georgia 110 Oxford court Athens, GA 30606 bmg1110@gmail.com Fudd Graham Auburn University 301 Funchess Hall Auburn, AL 36849-5413 grahalc@auburn.edu Michael Grodowitz USDA-ARS 59 Lee Road Stoneville, MS 38776 michael.grodowitz@ars.usda.gov Jennifer Henke Coachella Valley Mosquito & Vector Control District 43420 Trader Place Indio, CA 92201 jhenke@cvmvcd.org Bob Hickman BASF 1220 Manchester Road Maitland, FL 32751 robert.hickman@basf.com John Hopkins University of Arkansas Cooperative Extension Service 2301 S. University Ave Little Rock, AR 72204 jhopkins@uaex.edu Ellen Huckabay Alabama Cooperative Extension 302A Byrne Street Bay Minette, AL 36507 knighec@auburn.edu

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Richard Johnson USDA/APHIS/PPQ 4700 River Road, Unit 26 Riverdale, MD 20737 richard.n.johnson@aphis.usda.gov Phil Kaster KASCO MFG. & HERD SEEDER 170 W 600 N Shelbyville, IN 46176 INFO@KASCOMFG.COM Mallory Kelley Alabama Cooperative Extension System 2226 Hwy 14 West, Suite E Autaugaville, AL 36003 jones57@aces.edu Joseph LaForest Southern IPM Center 2360 Rainwater Road Tifton, GA 31793 laforest@uga.edu Nancy H. Y. Lee Chung Hsi Chemical Plant, Ltd. 4F, No. 20, Nanhai Road Taipei, Taiwan 100 master.first@msa.hinet.net Timothy Lockley Mississippi Department of Agriculture PO Box 5207 MS State, MS 39762 Tim@mdac.ms.gov Allis Wan-Cheng Lu Chung Hsi Chemical Plant, Ltd. 4F, No. 20, Nanhai Road Taipei, Taiwan 100 kang@chunghsi.com.tw

Joe MacGown Mississippi State University 100 Old Hwy 12 Mississippi State, MS 39762 jmacgown@entomology.msstate.edu Paul Mask ACES/AU 107 Comer Hall Auburn University, AL 36849 maskpau@aces.edu Carol Motloch USDA APHIS PPQ 903 San Jacinto Blvd, Suite 270 Austin, TX 78701 carol.m.motloch@aphis.usda.gov John Nabors AL Green Industry Training Center P.O. Box 383031 Birmingham, AL 35238 jtnabors@aces.edu Paul Nester Texas A&M AgriLife Extension Service 3033 Bear Creek Drive Houston, TX 77084 p-nester@tamu.edu Michael O'Shea Scotts Miracle-Gro 14111 Scottslawn Road Marysville, OH 43040 michael.oshea@scotts.com David Oi USDA-ARS, CMAVE 1600 SW 23rd Dr. Gainesville, FL 32608 david.oi@ars.usda.gov

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Jason Oliver Tennessee State University 472 Cadillac Lane McMinnville, TN 37110 joliver@tnstate.edu Kelly Palmer Alabama Cooperative Extension System 302A Byrne Street Bay Minette, AL 36507 ridleka@auburn.edu Manoj Pandey Tennessee State University 472 Cadillac Lane McMinnville, TN 37110 pandeymanoz@gmail.com Gabriela Perezchica-Harvey Coachella Valley Mosquito & Vector Control District 43420 Trader Place Indio, CA 92201 gharvey@cvmvcd.org Jeremy Pickens Auburn University PO Box 8276 Mobile, AL 36689 pickejm@auburn.edu Colleen Porter 2225 NW 36th Dr Gainesville, FL 32605 colleen_porter@cox.net Sanford Porter USDA-ARS, CMAVE 1600 SW 23rd Dr Gainesville, FL 32605 porters22@cox.net

Robert Puckett Texas A&M University AgriLife Extension 2556 F&B Rd College Station, TX 77843 rpuck@tamu.edu Darryl Ramoutar Scotts Miracle Gro 14111 Scottslawn Road Marysville, OH 43015 darryl.ramoutar@scotts.com Matthew Rawlings Scotts Miracle-Gro 14111 Scottslawn Rd Marysville, OH 43041 matthew.rawlings@scotts.com Dina Richman FMC Corp. 2929 Walnut Street Philadelphia, PA 19104 dina.richman@fmc.com Addie Rife The Scotts Miracle-Gro Company 14111 Scottslawn Road Marysville, OH 43041 addaleigh.rife@scotts.com Dennis Ring LSU AgCenter 404 Life sciences building 110 Union Square Baton Rouge, LA 70803 dring@agctr.lsu.edu Lawrence Shaw Orange County Mosquito & Vector Control District 13001 Garden Grove Blvd. Garden Grove, CA 92843 lshaw@ocvcd.org

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Jerry Sims Orange County Mosquito & Vector Control District 13001 Garden Grove Blvd Garden Grove, CA 92843 jsims@ocvcd.org Kerry Smith Alabama Extension Home Grounds 101 Funchess hall Auburn, AL 36849 smithkp@auburn.edu Jessica Strange-George MGK 8810 Tenth Ave N Minneapolis, MN 55427 jessica.strange-george@mgk.com Dan Suiter University of Georgia UGA Griffin Campus Department of Ent 1109 Experiment Street Griffin, GA 30223 dsuiter@uga.edu Melinda Sullivan USDA APHIS PPQ Field Operations 2150 Centre Avenue, Building B Fort Collins, CO 80526 melinda.j.sullivan@aphis.usda.gov Azieza Uhnavy Central Garden & Pet 1000 Parkwood Circle, Suite 700 Atlanta, GA 30339 auhnavy@central.com John Van Dyk FBA Consulting 42B Ellice Road Glenfield Auckland, New Zealand john@fbaconsulting.co.nz

Viv Van Dyk FBA Consulting 42B Ellice Road Glenfield Auckland, New Zealand viv@fbaconsulting.co.nz Doug VanGundy Central Life Sciences 12111 Ford Rd Dallas, TX 75234 repmn1@hotmail.com Robert Vander Meer USDA-ARS 1600 SW 23rd Drive Gainesville, FL 32608 bob.vandermeer@ars.usda.gov Antonette Walford Department of Agriculture and Water Resources PO Box 1006 Tullamarine, VIC, Australia 3043 antonette.walford@agriculture.gov.au Jason Williams University of Florida Entomology & Nematology Dept 6400 SW 20th Ave., Apt. #133 Gainesville, FL 32607 jwilli81@ufl.edu Nelson Wynn Regional Extension Agent PO Box 1606 Columbiana, AL 35051 wynnnel@aces.edu