1. POPULATION GENETICS OF GOPHER TORTOISES AT MOODY AIR FORCE BASE, GEORGIA David I. Mederos and J. Mitchell Lockhart Department of Biology Valdosta State University

2. The Gopher Tortoise (Gopherus polyphemus)

3. Gopherus polyphemus – Natural History  Found in Southeastern U.S. with major population concentrations in Florida, southern Georgia and Alabama  State Status – Threatened  Federal Status – Threatened in west AL, MS, and LA.

4. Photo Credit - LaShaun Lockhart

5. Gopherus polyphemus Gopherus polyphemus Quick Facts  Only tortoise east of Texas  Populations are concentrated in areas with deep sandy soils suitable for digging burrows  Live in loose colonies  Most fossorial of the North American tortoise species  40-80 year life span with 10-20 yr sexual maturity  ~1% recruitment rate

6. Gopherus polyphemus – Natural History Habitat preference: Habitat preference: Gopher tortoise’s are found in various habitats but require relatively well-drained soils for digging burrows, herbaceous food plants, and open sunny areas for resting and basking. Gopher tortoise’s are found in various habitats but require relatively well-drained soils for digging burrows, herbaceous food plants, and open sunny areas for resting and basking.

7. Gopherus polyphemus Considered a keystone species in the longleaf pine community, meaning its existence is CRITICAL to the existence of many other species. Considered a keystone species in the longleaf pine community, meaning its existence is CRITICAL to the existence of many other species.

8. The Gopher Tortoise Burrow

9. Part One: Gopher Tortoise Biology Part Two: Gopher Tortoise Restoration at Moody Air Force Base Part Three: Population Genetics Part Three: Uprising problems

10. Moody Air Force Base (MAFB) Photo Credits - Moody AFB

11. Moody AFB

12. Natural Resources Management Forestry Program Forestry Program 2,600 acres of upland pine/hardwood forests 2,600 acres of upland pine/hardwood forests 5,500 acres of bottomland forest/scrub-shrub 5,500 acres of bottomland forest/scrub-shrub Fish and Wildlife Program Fish and Wildlife Program Main Base hunting for military personnel Main Base hunting for military personnel Remainder of base is part of Grand Bay WMA Remainder of base is part of Grand Bay WMA Three installation lakes Three installation lakes Ecosystem Management Program Ecosystem Management Program Grand Bay-Banks Lake Ecosystem Council Grand Bay-Banks Lake Ecosystem Council RTE species management RTE species management

13. Moody Air Force Base Moody AFB is required by federal mandate to protect rare, threatened, and endangered (RTE) species likely to be impacted by the military mission of the installation. Moody AFB is required by federal mandate to protect rare, threatened, and endangered (RTE) species likely to be impacted by the military mission of the installation.

14. Moody Air Force Base 10,913 acres in Lowndes/Lanier Counties, Georgia. 10,913 acres in Lowndes/Lanier Counties, Georgia. 1. Main Base 1. Main Base Airfield and Cantonment areas Airfield and Cantonment areas Photo Credit - LaShaun Lockhart

15. Moody Air Force Base 2. Grand Bay Range, also serving as a state Wildlife Management Area (WMA) 2. Grand Bay Range, also serving as a state Wildlife Management Area (WMA) Weapons range and Buffer area Weapons range and Buffer area Photo Credit - LaShaun Lockhart

16. Part One: Gopher Tortoise Biology Part Two: Gopher Tortoise Restoration at Moody Air Force Base Part Three: Population Genetics Part Three: Uprising problems

17. Procedure Guanidinium Isothiocyanate extraction of nucleic acids Guanidinium Isothiocyanate extraction of nucleic acids - isolate DNA PCR – Epicentre PCR – Epicentre - FailSafe PCR PreMix worked best for multiplexing - Premix A and C USF-9 primers USF-9 primers - 2 multiplexed reactions - amplify 9 different VNTR loci  Gel electrophoresis - produce visible DNA fingerprint to analyze

18. Optimization

19. In-progress Results 71CPARCSBF 710.000000 CP0.0860100.000000 AR0.2910500.3095730.000000 CS0.2192970.1150570.4478960.000000 BF0.2660320.1739940.5031410.4174770.000000 Nei’s Genetic Distance Matrix Moody AFB Gopher Tortoise Populations Preliminary Analysis October 22, 2004 Note: Sample sizes are insufficient for a real, meaningful interpretation of the data. This is a trial analysis for the purpose of demonstration of capability.

20. Genetic Distance Tree Phylogenetic tree of Moody AFB Gopher tortoise populations. Trial analysis Note: Small population sizes prevent real interpretation of this tree. Analysis is for demonstration of capability. October 22, 2004 Continuous character Maximum Likelihood method version 3.61 +--------------------------------BF +---3 +----------------------2 +--------------CS +ACE- +ACE- +ACE- +-CP +ACE- 1-----------------------------------------AR +ACE- remember: this is an unrooted tree+ACE-

22. What can be obtained from these results? Genetic Distance Genetic Distance Migration- breeding Migration- breeding Are the colonies in fact separate or one big population Are the colonies in fact separate or one big population Importation of tortoises Importation of tortoises

23. Part One: Gopher Tortoise Biology Part Two: Gopher Tortoise Restoration at Moody Air Force Base Part Three: Population Genetics Part Three: Uprising problems

24. Upper Respiratory Tract Disease  Also called URTD’s  Causative Agent - Mycoplasma agassizii  Characterized externally by nasal and ocular discharge, palpebral edema, and conjunctivitis

25. Photo Credit - University of Florida Veterinary School

26. URTD’S and Gopher Tortoises Studies suggest that URTD’s negative tortoises are suitable for relocation and repatriation programs. Studies suggest that URTD’s negative tortoises are suitable for relocation and repatriation programs. Seropositive, clinically ill, culture or PCR positive animals should not be included in such programs. Seropositive, clinically ill, culture or PCR positive animals should not be included in such programs. One positive has been found, may be Imported animal One positive has been found, may be Imported animal Photo Credit - LaShaun Lockhart

27. Applications of Results o Land use changes o Relocation of Gopher Tortoises o Necessity of keeping infected tortoises separate from healthy tortoises o Which population possibly infected