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

POPULATION GENETICS POPULATION GENETICS OF GOPHER TORTOISES AT OF GOPHER TORTOISES AT MOODY AIR FORCE BASE, MOODY AIR FORCE BASE,

GEORGIAGEORGIA

David I. Mederos andDavid I. Mederos and

J. Mitchell LockhartJ. Mitchell LockhartDepartment of BiologyDepartment of Biology

Valdosta State UniversityValdosta State University

The Gopher TortoiseThe Gopher Tortoise((Gopherus polyphemusGopherus polyphemus))

Gopherus polyphemus – Gopherus polyphemus – Natural HistoryNatural History

Found in Southeastern Found in Southeastern U.S. with major U.S. with major population population concentrations in concentrations in Florida, southern Florida, southern Georgia and AlabamaGeorgia and Alabama

State Status – State Status – ThreatenedThreatened

Federal Status – Federal Status – Threatened in west AL, Threatened in west AL, MS, and LA.MS, and LA.

Photo Credit - LaShaun Lockhart

Gopherus polyphemusGopherus polyphemusQuick Facts

Only tortoise east of TexasOnly tortoise east of Texas Populations are concentrated in areas with Populations are concentrated in areas with

deep sandy soils suitable for digging burrowsdeep sandy soils suitable for digging burrows Live in loose coloniesLive in loose colonies Most fossorial of the North American tortoise Most fossorial of the North American tortoise

speciesspecies 40-80 year life span with 10-20 yr sexual 40-80 year life span with 10-20 yr sexual

maturitymaturity ~1% recruitment rate~1% recruitment rate

Gopherus polyphemus – Gopherus polyphemus – Natural HistoryNatural History

Habitat preference:Habitat preference: Gopher tortoise’s are found in various habitats but Gopher tortoise’s are found in various habitats but

require relatively well-drained soils for digging require relatively well-drained soils for digging burrows, herbaceous food plants, and open sunny burrows, herbaceous food plants, and open sunny areas for resting and basking.areas for resting and basking.

Gopherus polyphemusGopherus polyphemus

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

The Gopher Tortoise BurrowThe Gopher Tortoise Burrow

Part One: Gopher Tortoise BiologyPart One: Gopher Tortoise BiologyPart Two: Gopher Tortoise Part Two: Gopher Tortoise Restoration at Moody Air Force Restoration at Moody Air Force BaseBasePart Three: Population GeneticsPart Three: Population GeneticsPart Three: Uprising problemsPart Three: Uprising problems

Moody Air Force Base (MAFB)Moody Air Force Base (MAFB)

Rough Draft

Photo Credits - Moody AFB

Moody AFB

Natural ResourcesNatural ResourcesManagementManagement

Forestry ProgramForestry Program 2,600 acres of upland pine/hardwood forests2,600 acres of upland pine/hardwood forests 5,500 acres of bottomland forest/scrub-shrub5,500 acres of bottomland forest/scrub-shrub

Fish and Wildlife ProgramFish and Wildlife Program Main Base hunting for military personnelMain Base hunting for military personnel Remainder of base is part of Grand Bay WMARemainder of base is part of Grand Bay WMA Three installation lakesThree installation lakes

Ecosystem Management ProgramEcosystem Management Program Grand Bay-Banks Lake Ecosystem CouncilGrand Bay-Banks Lake Ecosystem Council RTE species managementRTE species management

Rough Draft

Moody Air Force BaseMoody Air Force Base

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


10,913 acres in Lowndes/Lanier Counties, 10,913 acres in Lowndes/Lanier Counties, Georgia.Georgia. 1. Main Base1. Main Base

Airfield and Airfield and Cantonment areasCantonment areas



2. Grand Bay Range, also serving as a state 2. Grand Bay Range, also serving as a state Wildlife Management Area (WMA)Wildlife Management Area (WMA) Weapons range and Buffer areaWeapons range and Buffer area



ProcedureProcedure Guanidinium Isothiocyanate extraction of nucleic acidsGuanidinium Isothiocyanate extraction of nucleic acids

- isolate DNA- isolate DNA

PCR – EpicentrePCR – Epicentre- FailSafe PCR PreMix worked best for multiplexing- FailSafe PCR PreMix worked best for multiplexing- Premix A and C- Premix A and C

USF-9 primersUSF-9 primers- 2 multiplexed reactions- 2 multiplexed reactions- amplify 9 different VNTR loci- amplify 9 different VNTR loci

Gel electrophoresis Gel electrophoresis - produce visible DNA fingerprint to analyze - produce visible DNA fingerprint to analyze

Optimization Optimization

In-progress ResultsIn-progress Results

71 CP AR CS BF

71 0.000000

CP 0.086010 0.000000

AR 0.291050 0.309573 0.000000

CS 0.219297 0.115057 0.447896 0.000000

BF 0.266032 0.173994 0.503141 0.417477 0.000000

Nei’s Genetic Distance Matrix

Moody AFB Gopher Tortoise PopulationsPreliminary AnalysisOctober 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.

Genetic Distance TreeGenetic Distance TreePhylogenetic tree of Moody AFB Gopher tortoise populations.Trial analysisNote: 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-

What can be obtained from these What can be obtained from these results?results?

Genetic DistanceGenetic Distance Migration- breedingMigration- breeding Are the colonies in fact separate or one big Are the colonies in fact separate or one big

populationpopulation Importation of tortoises Importation of tortoises


Upper Respiratory Tract DiseaseUpper Respiratory Tract Disease

Also called URTD’sAlso called URTD’s Causative Agent - Causative Agent - Mycoplasma agassiziiMycoplasma agassizii Characterized externally by nasal and ocular Characterized externally by nasal and ocular

discharge, palpebral edema, and conjunctivitisdischarge, palpebral edema, and conjunctivitis

Photo Credit - University of Florida Veterinary School

URTD’S and Gopher TortoisesURTD’S and Gopher Tortoises Studies suggest that URTD’s negative tortoises are Studies suggest that URTD’s negative tortoises are

suitable for relocation and repatriation programs.suitable for relocation and repatriation programs. Seropositive, clinicallySeropositive, clinically

ill, culture or PCRill, culture or PCRpositive animals positive animals should not be should not be included in such included in such programs.programs.

One positive has been found, may be Imported One positive has been found, may be Imported animalanimal


Applications of ResultsApplications of Results

o Land use changesLand use changeso Relocation of Gopher Relocation of Gopher

TortoisesTortoiseso Necessity of keeping Necessity of keeping

infected tortoises infected tortoises separate from healthy separate from healthy tortoisestortoises

o Which population Which population possibly infectedpossibly infected

Documents

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