Amphibian Decline - University Of Tennessee
Influences of Cattle Grazing onAmphibiansElizabeth Burton and Chandler SchmutzerUniversity of TennesseeDepartment of Forestry, Wildlife and FisheriesAmphibian Decline Climate changes Global warming UV-B rays Invasive species Competition / Predation Ie. Rana catesbeiana in the west Water contaminatesAmphibian Decline Pathogens Aeromonas hydrophila - “red leg”ChytridiomycosisIridovirusesRibeiroia1
Amphibian Declinesand Anthropogenic iological Conservation125: 75197219691966196301960 250Number of Populations Habitat DestructionNature404:752-755Influences of Cattle on Amphibians Previous researchHealey et al. 1997, Jansen and Healey 2002 -Australia Correlate amphibian abundance with wetland characteristics Suggest cattle indirectly negatively affect amphibiansBull et al. 2001, Bull and Hayes 2000 -Oregon Compare abundance of Columbia Spotted frogs in grazed and ungrazed areas Found no differences in abundance between treatmentsKnutson et al 2004- Minnesota Evaluate agricultural wetlands for value as amphibian habitat Species richness and abundance of some species lower at grazed wetlandsInfluences of Cattle on Amphibians Grazing vegetation Vegetation Structure Detritus Trampling egg masses Affects demographics at later life stages Soil Compaction Water Quality2
Influences of CattleDecrease in Water QualityDefecationNutrientsEutrophication Increases inammonia,nitrite, nitrateand phosphate Decrease inDissolvedOxygen Change in algalcommunityPlanorbid snailDecrease ingrowth andsurvivorship Change ininvertebratecompositionBird hostRibeiroiaP. llasnail sp.CercariaP. JohnsonP. JohnsonInfluences of gensDecrease WaterQualityIncreaseStressRanaviruses(FV3)Chytrid dIndividualMortality3
Amphibians and Cattle in TennesseeQuick FactsMost studies occur out west andalong streams(Belsky et al. 1999, Line 2002Jansen and Healey 2003) Amphibian richnesshighest in the southeast 44 anurans 84 caudates 40% of land area farmland 57% cattle production 48,000 cattle operations 9th in nation in beef cattle use Value of cattle 1.67 bill In Tennessee 21 anurans 45 caudatesBailey et al. 2006Redmond and Scott 1996NASS and USDAJustification It has been reported that cattle negatively affect emergent vegetation and water quality and thuscould potentially affect resident amphibians.Cattle could potentially increase pathogenoccurrence.The effect of cattle on adults has rarely beenquantified.There are no replicated studies for larvalamphibians.No studies documented in the Southeast,specifically Tennessee.Study AreaUniversity of TennesseePlateau Research and Education CenterCrossville, TN44AccesswetlandsNonaccesswetlandsAccess 10 yearsNever hadaccess28 March- 26August 200527 March- 25August 2006Size rangeAll pondshave fish0.153-1.29ha8 Wetlands4
Wetland 1Wetland 2Access WetlandsWetland 3Wetland 4Wetland 5Wetland 6Non-Access WetlandsWetland 7Wetland 8Larval Sampling DesignObjective 1. Determine the influence of cattle on larval amphibian abundance andrichnessNSeine plotDip net transectWater qualitymeasurementsiteRandomizedtransect foralgae, detritusand aquaticinvertebratesDip net transect Four Quadrants 2 Techniques Seine Plots Dip Net TransectsSeine plot1m5m10 m 4.5 m in lengthDip site every 1.5 m 3 sweeps at each dipsite 10 x 3 m permanentplot5
Larval SamplingObjective 1. Determine the influence of cattle on larval amphibianabundance and richness Larvae caught Counted Identified First 5 larvae per species Gosner stage (1960) recorded Measure BL and TL WeighedAny fish and invertebrates caught countedand identifiedGosner(1960)N Water Quality SamplingObjective 2. Determine the influence of cattle on waterSeine qualityplotDip net transect2.5 m from shoreWater variables measured at sampling location: Conductivity, temperature, pH and dissolved oxygenTurbidityWater qualityWater collected from sampling location measuredmeasurementfor:site Ammonia, nitrite, nitrate and phosphateRandomizedtransect foralgae, detritusand aquaticinvertebratesMeasured every 2 weeksSampling locationoccurred at 1 cardinalazimuthDip net transectYSI probeSeine plotLaMotte colorimeterFilamentous Algae, Detritus andInvertebrate SamplingFilamentousNDip net transectSeine plotObjective 3. Determine the influence of cattle on macroscopicfilamentous algae, detrital biomass and aquatic invertebrate abundance. Each Sample taken at a 0.5 m depth A cylinder (0.25-m2 ) placed in sample areaAll contents collected using a dip netRandomized (20 cm x 20 cm) Water qualitymeasurementsitetransect foralgae, detritusand aquaticinvertebratesContents collected and transportedon ice in plastic bags back to UTDip net transect1m5m10 mOnce per monthSeineplotMeasuredat one location in twoopposing quadrants in each wetland6
Filamentous Algae, Detritusand Invertebrate SamplingObjective 3. Determine the influence of cattle on macroscopicfilamentous algae, detrital biomass and aquatic invertebrateabundance. All samples sortedAlgae and detritus separated and dried at 80 Cfor 48 hours All invertebrates identified to family Weighed and dry mass recordedPathogen SamplingObjective 4. Determine the influence of cattle on the presence ofpathogens (viruses, bacteria and parasites) in larval communities, Pathogens measured Winter-February 15th 2005Summer-June 15th 2005Fall-October 12th 20052 species Bullfrog (Rana catesbiana)Green frog (Rana clamitans)SeiningLarvae collected opportunistically 5 individuals per species per wetlandDip-nettingBullfrogGreen frogRana catesbianaRana clamitansPathogen Processing MethodsObjective 4. Determine the influence of cattle on the presence of pathogens(viruses, bacteria and parasites) in larval communities, Transported back to UT Benzocaine hydrochloride Body mass and length, development stageGosner 1960 Fixed and fresh tissues UGA Veterinary Diagnostic andInvestigational Laboratory7
Pathogen SamplingObjective 4. Determine the influence of cattle on the presence of pathogens(viruses, bacteria and parasites) in larval communities, FV3 Identification Techniques: Virus isolation Electron microscopy PCRElectronmicroscopyof FV3Inoculationinto celllinesPCR of FV3MethodsMark Recapture(Objective 1. Determine the influence of cattle on species richness and relativeabundance of postmetamorphic amphibians) Pitfall Traps Fence surrounds 1/2 of thecircumference of each pond Buckets spaced every 5 or 10mwith separating leads Electric fence (access ponds) Pitfalls opened 2X per week Pitfalls opened for 24h thencheckedProcessing Captured Individuals Measure (SVL) Weigh Tag-VIA tags Mark-Toe clipping8
MethodsBreeding Call SurveysObjective 1. Determine the influence of cattle on species richness and relative abundance ofpostmetamorphic amphibiansSurveys followed North American AmphibianMonitoring Program (NAAMP) protocol 2 survey durations 5 minutes (0-5:00) 10 minutes (0-10:00) 2 Permanent listening stationsObservers did not share survey resultsExposed to the same chorus Began 30 minute after sunset Upon arrival waited 1 minute Species occurrence and ranked abundanceMethods Ranking species-specific abundance 1 individuals can be distinguishedand calls do not overlap 2 individuals can be distinguishedand calls do overlap 3 full chorus (individuals cannot bedistinguished and calls do overlap)MethodsSampling TechniquesObjective 2: Determine the influence of cattle on egg mass abundance Egg Mass Surveys Each wetland visuallysurveyed once perweek Traverse a permanentbelt transect in 2randomly selectedopposing quadrants Transects are 10 mlong and 2 m from andparallel to shore9
MethodsSampling TechniquesObjective 2: Determine the influence of cattle on shoreline vegetation structure and composition Vegetation Structure and Height Measured with graduated profile board Percent Horizontal Cover Visual estimation of a 1-m2 plot Plant Species Richness Enumerated in a 1-m2 plotMeasured once per monthPlot location: midpoint of vegetation zonealong random azimuth in 2 opposingquadrantsMethodsSampling TechniquesObjective 4. Determine the influence of cattle on pathogen and malformation prevalence in amphibians Pathogen prevalence 5 metaporphs Rana clamatins collected from each wetland onJune 15, 2005 Individuals euthanized via transdermal exposure to benzocainehydrocloride Comprehensive histological and parasitological analysis of tissuesamples performed at the Tifton Veterinary Diagnostic andInvestigational Lab Bacteria, viruses, parasites and other pathogensMethodsSampling TechniquesObjective 4. Determine the influence of cattle on pathogen and malformation prevalence in amphibians Trematode prevalence Malformed individuals opportunistically collected Malformation classified using USGS Field Guide to Malformations ofFrogs and Toads Humanely euthanized via transdermal exposure to benzocainehydrochloride Fixed in 10% buffered formalin and Cleared Light microscopy used to detect presence of encysted trematodemetacercariae10
Statistical AnalysesRepeated Measures ANOVA: Amphibians Response: Relative Daily Abundance Effects: Access Treatment, MonthTwo-sample T-tests (Trt*Month, P 0.1)Repeated Measures ANOVA: Egg Mass Response: Mean Total Abundance Effect: Access Treatment, MonthStatistical AnalysesRepeated Measures ANOVA: Vegetation Response: Mean Vegetation Structure Vegetation Variables: Percent Vertical &Horizontal Cover, Height Effects: Access Treatment, MonthTwo Sample Z-test: Pathogens and MalformationsResultsLarval Abundance12.00BDaily relative abundance10.008.00RACA 2005RACL 20062.9X greater5X FOPSCRRACARACLRAPASpeciesAll otherp 0.11α 0.10SAS AnalysisRepeated measures ANOVA11
ResultsLarval Diversity and RichnessBp 0.080.073-4Xgreater0.060.4A0.3p 0.130.0820060.5A0.092.7X greater0.6D iv e rs ityR e la tiv e da ily s pe c ie s ric hne s atmentAnalysisDiversity Shannon-Weiner Diversity Index Algorithmα 0.10SAS Repeated Measures ANOVAResultsSpecies Composition1.000.90Proportion of Species0.80s 9s essNon-accessTreatmentCricketfrogGreen frogLarval Species CapturedSpringPeeperGrey treefrogBullfrogMudpuppySouthernLeopard FrogEastern red-spotted newtPickerel frog12
ResultsFish Composition1.000.90Proportion of aredSunfishNon-accessLargemouth BassFish Species CapturedBluegillGolden ShinerGreen SunfishResultsWater Quality DO 28.2% nonaccess (2006)S p ecific Co n d u ctivity (m S cm -1)120A87B8060402001TreatmentAp 0.1554 NH3, NO2, NO3and TEMP321 Higher in access01Treatmentp 0.09100 All other waterquality variablesBp 0.036AnalysisRepeatedMeasuresANOVAα 0.10SAS T u rb i d i ty l e v e l s (F o rm a z i n T u r b i d i ty U n i ts )1409D is s olv e d O x y ge n le v e ls (m g/L) SPCOND 67.8% in access (2005)70.4% in access(2006) TURB 3.7X access (2005)3.5X access(2006)10090Ap 0.058070B60504030201001Treatment13
ResultsAlgae and Detritus7p 0.35p 0.09140120Biomass in grams5Biomass in ess60A402000Algae-1Detritus 10.9X non-access in 2005Analysisα 0.10SAS Repeated Measures ANOVAResultsAll other pvalues wereInvertebrate Abundancep 0.1140.000A35.000LIBE: 1.8X (2005) and 5.2X (2006) non-accessA25.000OLIG: 4.9X (2005) accessPLAN: access15.000PHYRelative abundance30.000Taxaα 0.10SAS AnalysisRepeated measures ANOVADragonflylarvaeLIBESmall SquaregillMayflyNon-BitingMidge atesCapturedDale ParkerDale ParkerSWCSMHAquaticwormOLIGPlanorbidsnailPLANPea ClamPhysid SnailPHYSSPHALeechHIRU14
ResultsRegression Model2005 BUFO 0.098 (TURB) – 5.076 (NH3)RAPA 0.004 (OFISH)2006 RACA 0.393 (OFISH)RACL – 0.026 (SPCOND) 0.556 (SR)RAPA 0.001 (PREDF)standardized coefficients presentedResultsPathogen PrevalenceTreatment0.5P 0.78AAFV3 Prevalence0.4P 0.020.40.363.9XMoreLikely!!!A0.30.3B0.20.15Cattle LandUseAccessNon-access0.10BullfrogGreen Frogn 104 tadpolesn 80 tadpolesAnalysisLogistic Regression and Maximum Likelihood EstimationResultsPathogen PrevalenceFV3 PrevalenceSeasonal Effects0.7A0.60.57P 0.02P 0.006B0.57.7X0.4More Likely!!0.30.2BB0.240.454.7X MoreLikely!A0.150.1Trt*SeasonDid notInteract,P 0.30SeasonW interSummerFall0.15No WinterCaptures0BullfrogGreen Frogn 104 tadpolesn 80 tadpolesAnalysisLogit and Logistic Regressions and Maximum Likelihood Estimation15
ResultsDevelopmental Stage EffectBullfrog0.70FV3 Prevelance0.600.550.50Bullfrogs28% Decrease in the PredictedOdds of Infection0.50with each unit increase inGosner stage.0.380.40n 102 9-3031-33P 0.00534-3536-3738-394041Gosner StageGosner Stage(1960)AnalysisLogistic Regression and Odds-Ratio EstimatesResultsDevelopmental Stage EffectGreen Frog0.70Green FrogsNo detectable trend.n 80 tadpolesFV3 4041Gosner StageP 0.872Gosner Stage(1960)Summary of Results Larval Abundance, Richness and Diversity Bullfrog and green frog abundance was greater in nonaccessSpecies richness was greater in non-access wetlandsNo significant difference in species diversityWater Quality Specific conductivity and turbidity were higher and dissolvedoxygen lower in cattle-access wetlandsNo significant difference in other water quality variablesDetritus and Algae Detritus was greater in non-access wetlandsNo significant difference in algae biomass betweentreatments16
Summary of Results Invertebrates Regression Model Dragonfly larvae abundance was greater in non-accessAquatic worm abundance was greater in cattle-accessSpecific conductivity explained 82% of variation in green frog larvalabundance.Other fish (non-predators) explained 73% of variation in bullfroglarval abundance.FV3 Green frog larvae were more likely to be infected with FV3 in cattleaccess wetlands.FV3 prevalence was higher in cooler months for both species.As development progressed FV3 prevalence decreased inAmerican Bullfrog larvae.Discussion Larval Abundance It was documented that cattle access wetlandsnegatively impacted American bullfrog and greenfrog tadpole populations Water quality and fish abundance were important predictorsof abundance It appears that American and Fowler’s toadtadpoles were not negatively impacted by cattleaccess Higher resistance to water quality Jofre and Karasov 1999 Exploitation of habitat where there is lower abundance ofranids DiscussionDetritus and Algae Detritus in non-access wetlands Algae trend toward being in cattle-access wetlands LIBE – somewhat tolerantOLIG –tolerant to water pollutionIn General In-direct effects from lack of grazing pressure Provided better habitat for ranids Trend toward higher nutrientsInvertebratesVoshell 2002 More snails in cattle-access Slight change in composition – difference in abundance More “sensitive” species in non-access wetlands17
Discussion Water quality Cattle negatively impacted water quality Reductions in water quality can increasemortality of amphibian eggs and larvae Induces stress making them moresusceptible to pathogensBoyer andGrue 1999Carey et al. 1999Discussion Pathogen Prevalence- Frog Virus 3 (FV3) Water quality Effected green frog survival Potentially compromised immunityJofre and Karasov 1999 Seasonal Effects Low temperatures increases pathogen prevalence Low temperatures cause a decrease in overall immunefunction Developmental StageManiero and Carey 1997 Immunity could increase in bullfrogs Brunner et al., 2004 Susceptible tadpoles at earlier stages experiencedmortalityGosnerStage(1960)18
Desmognathus ocoeeAmbystoma talpoideumNotophthalmusviridescensPseudotriton montanusPlethodon glutinosusResultsPostmetamorphsSpecies Specific Relative Abundance of Pitfall Trap CapturesP 0.06P 0.198.4X in 20062.3X in cies Specific Relative Abundance of Pitfall Trap CapturesP 0.06P 0.189.8X in 20062.5X in 200519
Results Species Richness did not differ between treatments Species Diversity did not differ between treatmentsSpecies CompositionS 12S 13ResultsEgg Masses1.8AMean number of egg masses1.61.41.2P 0.3510.80.60.4A0.20Acce ssNon-acce ssTreatmentResultsVegetation ResponsesA cce ssHeight (m)Access10.90.80.70.60.50.40.30.20.10Non-acce 05074% 200641% 2005BAAccessNon- accessPercent Vertical StructurePercentVertical StructurePercent Horizontal CoverP ercent Vertical essPercentHorizontalCPercentHorizontalCover20
Results: Pathogens No differences in bacterial, viral or parasitic prevalencebetween treatments 35 malformed individuals 11 Malformation types2% malformation rateAnophthalmiaPolymeliaPercent Malform ations706050AccessPolydactylyAP -Access41%20102 individuals withtrematode metacercariae0AccessNon-AccessTreatmentSummary of Results Green frog metamorphs are negatively affected by cattlepresence Vegetation structure, horizontal cover and height are reducedin cattle access wetlands Egg mass abundance did not differ between treatments Prevalence of pathogens and malformations did not differbetween treatmentsDiscussion Vegetation Structure: Breeding sites Foraging and escape cover Egg depositionJansen & Healey (2003),Healey et al. (1997) Green Frog Metamorphs driving the trends Higher Abundance More sensitive to disturbance Heavily impacted by vegetative cover and water quality Toads- going against the trend Reduced Competition / Out competing other species Better suited to areas with less vegetation Desiccation, movement Can withstand lower water quality than other species21
Discussion Water ccess Reduced immunocompetenceincreased FV3 prevalence Tadpoles affect later demographic stages Modeling Postmetamorphic Amphibian AbundanceEnvironmental Cofactors: vegetation, water quality, cattle density, tadpole abundance2006 RACL -0.002(Specific Conductivity)R2adj 0.7982% Specific Conductivity2005 BUAM 0.0004(Cattle) – 0.39(NO2) – 0.0002(Turbidity) – 0.03(P04)R2adj 0.9983% Cattle Density2006 BUAM 0.0001(Turbidity) – 0.006(NH3) 0.002(Temperature)R2adj 0.9990% TurbidityConservation Implications Cattle grazing may be contributing toamphibian declines Separation of cattle and amphibians Providing alternative food and watersourcesAcknowledgementsFunding: UT Dept. of Forestry, Wildlife and Fisheries Tennessee Wildlife Resources AgencyAssistance: Walt Hitch PREC Staff Volunteers22
4 Amphibians and Cattle in Tennessee Quick Facts 40% of land area farmland 57% cattle production 48,000 cattle operations 9th in nation in beef cattle use Value of cattle 1.67 bill NASS and USDA Amphibian richness highest in the southeast 44 anurans 84 caudates In Tennessee 21 anurans 45 caudates Redmond and Scott 1996 Most studies occur out west and
2015 BP Amphibian Egg Mass Survey Results 4 The egg masses and/or adults of five species of amphibian were identified within the study area (Table 1). All the species detected were those that breed in aquatic habitats. No terrestrial amphibian species were detected, but none were expected as the survey protocol focused on aquatic habitats. Table 1.
The SLT amphibian monitoring program is accomplished with the help of new and past citizen science volunteers and Land Stewards. Amphibian identification training, adapted from the Whatcom Amphibian Monitoring Project, was conducted on February 4, 2017 to teach and refresh the materials and methods used to conduct amphibian egg mass surveys.
Burke Lake Reptile & Amphibian Merit Badge bout how to prepare your scout for Reptile & Amphibian Merit Badge Program at Burke Lake Park. After reading information, print out Reptile & Amphibian Workbook (pages 5 -19 of PDF) to bring to class. Be sure that you are familiar with the requirements for th
78 REPTILE AND AMPHIBIAN STUDY Reptile and Amphibian Study Resources. Ross, Charles A. Crocodiles and Alligators. Facts on File, 1989. Zug, George R., Carl H. Ernst, et al. Smithsonian Answer Book: Snakes, 2nd ed. Smithsonian Books, 2015. Caring for Reptiles and Amphibians in Captivity Bartlett, R.D. The 25 Best Reptile and Amphibian Pets.
2.1. Decline Curve Analysis. The Arps decline curve is the most common DCA. The Arps hyperbolic decline curve model is [29]. q q i ðÞ1 bD i t 1/b, ð1Þ where q is the predicted production, q i is the initial produc-tion, t is time, b is a constant, and D i is the initial decline rate. When the constant loss ratio b is 0, the decline curve
summarizes the observations for this pool. One large amphibian egg mass was observed on April 14, 2005. During that visit, a local home owner mentioned that spring peepers (a facultative amphibian species) were calling at night. By April 26, 2005, numerous amphibian egg masses
AMPHIBIAN AND REPTILE DISEASES MATTHEW J. GRAY* Department of Forestry, Wildlife and Fisheries, Center for Wildlife Health, University of Tennessee Institute of Agriculture, Knoxville, Tennessee 37996, USA AMANDA L. J. DUFFUS Department of Biology and Physical Sciences, Gordon State College, Barnesville, Georgia 30204, USA KATHERINE H. HAMAN
English Language Arts & Literacy in History/Social Studies, Science, and Technical Subjects ISBN 978-0-8011-17 40-4 . ISBN 978-0-8011-1740-4. Bar code to be printed here. California Common Core State Standards. English Language Arts & Literacy in . History/Social Studies, Science, and Technical Subjects. Adopted by the. California State Board of Education . August 2010 and modified March 2013 .
