Historical Perspective

1. The American Burying Beetle in Ohio: Current Status and Future Direction of the 1st Mainland Reintroduction and Ohio’s Captive Rearing Program.Presented by George D. Keeney and David J. Horn The Ohio State University Department of Entomology

2. Historical Perspective • Last known extant occurrence in Ohio at Old Man’s Cave in Hocking Hills State Park (1974). • Collected from dead woodchuck. • Found in OARDC collection. • The first mainland reintroductionof ABB to historic habitat

3. Last known historical occurrence of ABB in Ohio (1974, Hocking Hills St. Park.) OH counties where ABB is currently listed

4. The Search for Extant Populations • Started surveying in 1988. • Obtained small grant from ODNR in 1993. • Considered Lake Erie islands, Wayne National Forest, various ODNR forests, associated state parks, wildlife areas and metro parks.

5. Considerations in surveys and selection of release sites • Land use/ownership. • Types and numbers of congenerics. • Degree of scavenger disturbance. • Estimated type, weight and amounts of carrion available. • Other rare or unusual fauna

7. Ohio Recovery Efforts: • Restoration Goals and Objectives • To establish a self-sustaining viable population of the American burying beetle within Ohio; • To maintain at least one captive breeding colony of ABBs to provide adequate numbers of animals for release; and • To produce and distribute informational materials for the general public and to conduct educational programs about the ABB.

8. Waterloo Wildlife Area and Experimental Station Adjoins 26,827 acre Zaleski St. Forest, Lake Hope St. Park and Wayne National Forest OH counties where ABB is listed

10. Summary of ABB Releases for Reintroduction to Ohio

11. The Wilds ConservationArea -14 square miles of primarily grassland - Reclaimed strip mine land - 200 acres of wetlands - 150 lakes

12. The OSU Biological Control Quarantine Facility formerly housed our captive breeding colony.

13. Brooding our ABB • 14 hour photoperiod • 25° Celsius

14. Adult beetle maintainance • 14 hour photoperiod • 25° Celsius • held here 3-4 weeks before cooler storage or breeding.

15. Adults are housed separately in 3”w x 3”h x 7”l crisper. • Adult beetles are fed combination of Galleria and Tenebrio larvae. • Damp paper toweling changed every 3-4 days.

16. Evening approaches!

17. 2007 Action Items • Continue surveying effort… • Intensify surveys at Waterloo Wildlife Area in spring prior to ABB release for 2007. • Expand survey area by utilizing smaller “sentinel” surveys in other tri-county areas, i.e. state forest land, state parks, national forest, private land, etc. • If ABB are found in a sentinel area, then focus more effort in that locality.

18. 2007 Action Items • Continue surveying effort… • Continue with 3rd year of surveys at The Wilds. • Prepare for release perhaps in 2008? • Continue with surveys in the nearby Wayne National Forest. • Long Hollow (Athens County) and Wildcat Hollow (Perry County). • Tick Ridge (Vinton County)? • Adjoining private lands of interested participants?

19. 2007 Action Items • Placement of road kill at several specific sites in Waterloo Wildlife Area away from release areas. • Provide carrion resources as an adult ABB feeding resource. • May attract and hold ABB in the general vicinity. • Easily checked for Nicrophorus spp. at any visit without setting up transects. • Perhaps satiating local vertebrate scavengers or throwing them “off the scent”? Perhaps “low-hanging fruit”?

20. 2007 Action Items • Monitor the fate of carrion at Waterloo Wildlife Area. • Determine how carrion is utilized when available in “protected” and “unprotected” conditions? • Is it being used by Nicrophorus spp.? Which ones? • Effective in detecting ABB compared to baited transects? • Determine type and numbers of vertebrate scavengers? • Feral pigs, coyotes, turkey vultures, raccoon, etc. • Evaluate protective measures prior to 2007 release.

21. 2007 Action Items • Tools in determining type and numbers of vertebrate scavengers. • Infrared cameras trained on carrion. • Road kill. • “Protected” and “unprotected” quail carcasses. • “Wet sand” prints? • Correct for any deficiencies in protective measures against vertebrate scavengers prior to 2007 release.

22. 2007 Action Items • Release of ABB supplied by the St. Louis Zoo • Release is scheduled for Thursday, June 21, 2007. • Estimated that 100 or so pairs of captive reared beetles will be provided. • Will be last year of releases at Waterloo Wildlife Area. • The bucket rearing method which was utilized last year on a limited trial basis will be expanded. • Rolled heavy duty garden fence will probably be utilized in lieu of poultry netting squares to protect brooding ABB after burial.

23. 2007 Action Items • Post-release ABB activities • Monitor burial and brooding success of ABB 10 to 15 days post-release (weather dependent). • Estimate potential brood size by sampling. • Determine impact of scavengers. • Provide potential “alternative” brooding opportunities locally if adults abscond from those provided. Maybe useful in monitoring.

24. 2007 Action Items • Post-release ABB activities • Continue to closely monitor Waterloo specifically throughout season. • Continue to conduct surveys throughout greater Athens/Hocking/Vinton area. • Provide supplemental carrion in the general locality to emerging beetles later in season. • Local control measures against feral pigs may be implemented by ODNR on Waterloo as this is also a wild turkey management area.

25. 2007 Action Items • The captive ABB breeding program in Ohio • A few pair of adults from the arriving St. Louis stock will be initially culled to establish a small captive colony at The Wilds. • A few pair of adults produced from the buckets used at the Waterloo release site will be culled upon emergence to reestablish a captive colony at The Ohio State University Department of Entomology.

26. Future Direction • Captive cultures reestablished with beetles from St. Louis Zoo • A few pairs culled initially for establishment of The Wilds colony • Field reared beetles culled later for OSU colony • Increase numbers of captive reared ABB at The Wilds and OSU in 2007 • Involvement by Cincinnati Zoo as a tertiary captive population (2008?)

27. Justifications for multiple rearing sites • Increases ability to raise more quality beetles by… • Effectively increasing manpower • Increasing rearing space • Spreading burden of limited financial, physical and personnel resources • “Puts the eggs in more than one basket” • Increases potential genetic diversity by periodically outcrossing between cultures

28. Future home of OSU ABB colony • Access to water, walk-in cooler, environmental chambers • Adequate space for brooding, maintaining adults • Controlled stable environment • Interior room • Temperature, ventilation and lighting controlled • Secure

29. Future Direction • Releases planned for The Wilds in 2008 or 2009 using ABB reared at The Wilds and OSU? • Additional releases planned at The Wilds and perhaps additional sites in SE Ohio beyond 2008 or 2009? • Study on fate of carcasses

30. Current release sites = Waterloo Wildlife Area =Potential future release sites The Wilds Wildcat Hollow (WNF) Long Hollow (WNF)

31. Assessment of the Potential Impact of Chlorophacinone on Burying Beetles. • Introduction: • Chlorophacinone is an indandione anticoagulant rodenticide. • Used to control nuisance prairie dogs, pocket gophers and voles. (OK, KS and NB). • Potential impact of this chemical on ABB needed to be assessed. • Nicrophorus orbicollis, was used as a surrogate species. • Nicrophorus orbicollis = nearest extant relative to ABB.

32. Assessment of the Potential Impact of Chlorophacinone on Burying Beetles. • Hypothesis/Expectations: • Exposure to chlorophacinone would have minimal to no impact on beetle behavior or survivorship. • No direct acute mortality effects to adult beetles and/or immature forms. • No direct acute/chronic effects upon adult behavior and/or immature forms. • No indirect or secondary effects upon adults and/or immature forms, i.e. reduced fecundity of adults, issues with immature growth and development, etc.

33. METHODS/MATERIALSPhase 1.) Investigation of any acute or chronic effects of chlorophacinone upon Nicrophorus larvae developing in dosed carcasses and the subsequent generation of adults produced from those carcasses • Test carcasses of female laboratory Norway rats. • Treatment: Fed exclusively 50 ppm chlorophacinone bait for a period of 5 to 10 days. • Control: Undosed rats fed upon a standard laboratory rodent diet. • Standard procedures used for production of ABB. • The total number of young produced per brood was tallied, sexed and weighed.

34. METHODS/MATERIALSPhase 2.) Investigation of any direct acute or chronic effects of chlorophacinone upon Nicrophorus adults and the subsequent ability of those adults to brood and produce normal progeny • Adult beetles (128 total) were maintained for 28 days on a no-choice diet. • Treatment: Homogenized ground beef containing 3.0 ppm chlorophacinone. • Control: Non-dosed homogenized ground beef. • Beetles were observed daily for signs of aberrant behavior and scored for mortality.

35. METHODS/MATERIALSPhase 2.) Investigation of any direct acute or chronic effects of chlorophacinone upon Nicrophorus adults and the subsequent ability of those adults to brood and produce normal progeny • After the trial period, were then offered Japanese quail carcasses. • Standard procedures used for production of ABB. • Treatment beetles were closely observed for aberrant behavior during carcass burial, preparation and brooding. • The total number of young produced per brood were tallied, sexed and weighed.

36. RESULTS/DISCUSSIONVariables measured • total number of beetles produced/brood, • total number of beetles produced/carcass weight, • weight/beetle, • estimated total brood weight/brood, • total brood weight/carcass weight, • total numbers of females/brood, • total number of males/brood, • proportion of males/brood.

37. RESULTS/DISCUSSIONPhase 1.) Investigation of any acute or chronic effects of chlorophacinone upon Nicrophorus larvae developing in dosed carcasses and the subsequent generation of adults produced from those carcasses • Significant differences existed in rat carcass weights between treatment and control. • Difference was reflected in the raw unadjusted data. • total number of beetles produced per brood. • total brood weight. • Transforming data or adjusting for carcass weight. • Statistically significant differences disappeared with the exception of mean number of females produced per brood. • Difference may be attributable to average carcass weights between treatment and control .

38. RESULTS/DISCUSSIONPhase 2.) Investigation of any direct acute or chronic effects of chlorophacinone upon Nicrophorus adults and the subsequent ability of those adults to brood and produce normal progeny • Over the duration of the 28 day trial feeding period using the no-choice diets • 12.5 % of the control adults died (9.4% female/3.1% male). • 4.7% of the treatment adults died (1.6% female/3.1% male). • Chlorophacinone has no direct negative impact on acute mortality of adults.

39. RESULTS/DISCUSSIONPhase 2.) Investigation of any direct acute or chronic effects of chlorophacinone upon Nicrophorus adults and the subsequent ability of those adults to brood and produce normal progeny • In the second aspect of this experimental phase, the distribution of quail carcass weight was carefully held to a relatively narrow range. • No significant differences occurred for any of the variables measured. • It is believed that the lower number of brood produced in this experiment, relative to the first, which used rats as the carrion resource, may be attributed to either the age of the beetles and/or the use of a different type/condition of carrion.

40. ConclusionsChlorophacinone would appear to have… • No negative impact on the survival, burial or brooding of adult N. orbicollis. • No negative impact or the growth and development of their larvae when carcasses contaminated with chlorophacinone are used as breeding resources. • No direct negative impact on acute mortality of the adults. • No apparent indirect or secondary effects upon adults and/or immature forms. • By implication, no impact on ABB, since the biology of ABB is so similar to that of N. orbicollis.

41. Acknowledgements We wish to thank the numerous individuals from the United States Fish and Wildlife Service, the Ohio Department of Natural Resources Division of Wildlife, the Oklahoma Biological Survey, the St. Louis Zoo, Fort Chaffee, and others from The Ohio State University, for their wonderful efforts in support of this program.

42. Acknowledgments • ODNR Division of Wildlife: Carolyn Caldwell, Dave Swanson • USFWS Endangered Species: Sarena Selbo, Angela Zimmerman, Mike Amaral • University of Oklahoma: Kiki Hiott, Gary Schnell • The Ohio State University: Foster Purrington, Carrie Fisher, Dick Maxey, Matt Schroeder, Joshua Bryant • The Wilds: Adam Davis, Steve Shurter, Evan Blumer • Saint Louis Zoo: Jane Stevens, Bob Merz

43. Acknowledgements This program is supported by funds from the Ohio Department of Natural Resources Division of Wildlife “Do Something Wild” tax check-off program.