A s amphibian biologists bearing witness to the sixth mass extinction, we have to

A s amphibian biologists bearing witness to the sixth mass extinction, we have to

be careful not to lock ourselves down into some form of pre-clinical post-traumatic stress disorder. The world, whether it realizes it or not, needs us. The trick in all this is to not lament the loss, but to get on with it, and to find energy; and rather than mourn what is no longer, try to celebrate those things that are or were. Easier said than done, and for people in certain sub- professions of our field this can be more than difficult. My friends who work in zoos and aquariums, especially those who run captive breeding programs, are particularly vulnerable. Their jobs are high stress—they fail to succeed and there are forever consequences. I understand their loss of hope, which can sometimes descend deeper, into a place that is much darker and from where it is more difficult to rise. These are people who, if they drink, drink faster and more than your average herpetologist, which is saying something. What our friends need to understand is that the problem does not lie with them; it lies within the situation they find themselves in. If roles were reversed it would be us on the floor next to that barstool. We are asking them to be on the front lines of the sixth mass extinction, and we’re not giving them the financial or emotional support they need. They are frazzled and fraying. It is easy for them to think they’re alone, and following a loss, it’s easy for them to despair. But their fight is our fight, and they need our help. Why wouldn’t we expect burnout

No Retreat, Baby, No Surrender.

Michael J . Lannoo

and despair, and the sorts of behaviors isolated on mountain fragments.

in bars typically reserved for returning If substantiated,

taxonomic

soldiers and off-shift firefighters? These are revision will be necessary with people that need to know they’re not alone, implications for conservation and that they have our support. Bruce assessments. The two sampled Springsteen understood when he wrote: lowland, coastal individuals are

nested within primarily montane

“Yes, we made a promise we swore we’d clades. Molecular clock estimates always remember.

No retreat, baby, no surrender.”

Blood brothers in the stormy night a vow Boulengerula, perhaps suggesting to defend.

regional causal events. However,

No retreat, baby, no surrender.”

other divergences happened at different times and over a

Columbia Records, 1984

long period, perhaps extending back to the Oligocene/ Eocene.

Fu ll a rticle : La n n o o , M.J. 2 0 11.

Our results for Boulengerula

N o Re tre a t, Ba by, N o Su rre n d e r.

suggest a role for relative long-

H e rp e to lo gica l Re vie w 4 2 :14 2 – 14 5 .

term environmental stability in the origins of the Eastern Arc Mountains biodiversity hotspot.

Molecular phylogenetics of Boulengerula (Amphibia:

Fu ll a rticle : Lo a d e r, SP,

Metamorphic (top) and paedomorphic (bottom) Oklahoma

salamanders ( Gymnophiona: Caeciliidae) Eurycea tynerensis). Images by Samuel D. Martin. W ilkin s o n , M., Co tto n , J.,

and implications for taxonomy,

Me a s e y, G.J., Me n e go n ,

biogeography and conservation

M., H o w e ll, KM., Mü lle r,

and conservation attention focused only

on paedomorphic populations. We perform Simon P. Loader, Mark Wilkinson, J ames A.

H . Go w e r, D J. 2 0 11. Mo le cu la r

phylogenetic analyses of mitochondrial and M. Howell, Hendrik Müller & David J . Gower

Cotton, G. J ohn Measey, Michele Menegon, Kim

p h ylo ge n e tics o f Bo u le n g e r u la

( Am p h ibia : Gym n o p h io n a :

nuclear genes to assess patterns of historical

Ca e ciliid a e ) a n d im p lica tio n s

isolation in

E. tynerensis, and test whether

T life history mode is randomly distributed

he African caecilians of the genus Boulengerula are almost entirely

fo r ta xo n o m y, bio ge o gra p h y a n d

with respect to the phylogeny and geography. confined to the Lowland coastal and Eastern

We find three divergent mitochondrial Afromontane region but little is known of

co n s e rva tio n . H e rp e to lo gica l

Jo u rn a l, 2 1: 5 -16 .

lineages and significant shifts in allele their evolutionary history. Phylogenetic

frequencies of a nuclear gene between the relationships of the East African caeciliid

eastern, western, and southwestern portions Boulengerula were investigated using

Considering alternative life history

of the distribution. Life history mode mitochondrial sequence data from Kenya

modes and genetic divergence in

varies extensively, but paedomorphosis and Tanzania. The phylogenetic tree

is largely restricted to the widespread supports a formerly proposed bipartition

conservation: a case study of the

western clade. Therefore, the two most of the genus, and differs significantly from

Oklahoma salamander

divergent and narrowly distributed clades recent morphological analyses. Our analysis

Sarah L. Emel & Ronald M. Bonett

(southwestern and eastern) were previously identified genetic differences between

A provide important variation for the long-term persistence of a species. However,

lternative life history strategies can

overlooked due to their metamorphic life several mtDNA groups that potentially

history. Paedomorphosis has allowed E. represent undescribed species – populations

conservation of such lineages

tynerensis to drastically increase its niche

can be complicated because

breadth and distribution size. Nevertheless,

each life history mode may have

metamorphosis is also an important

different habitat requirements

attribute, and metamorphic populations

and may be vulnerable to

are the ultimate source for paedomorphic

different

environmental

evolution. Preservation of divergent genetic

lineages, and regions that include adjacent

to identify and conserve

habitat for both life history modes, may be

divergent intraspecific genetic

the most effective way to maintain historical

lineages of amphibians is well

and adaptive variation and provide

recognized, but conservation of

gateways for ongoing life history evolution.

alternative life history modes has been underemphasized. Fu ll a rticle : Em e l, S. L., a n d R. M.

The Oklahoma salamander Bo n e tt. 2 0 11. Co n s id e rin g a lte rn a tive ( Eurycea

tynerensis) life h is to ry m o d e s a n d ge n e tic endemic d ive rge n ce in co n s e rva tio n : a ca s e to the Ozark Plateau of North America, s tu d y o f th e Okla h o m a s a la m a n d e r.

is

and has two discrete life D OI 10 .10 0 7/ history modes, metamorphic s 10 5 9 2 -0 11-0 2 2 6 -9

recently, these modes were considered separate species

Boulengerula. Photo: Michele Menegon

56 | FrogLog Vol. 97 | July 20 11

Conservation genetics of

gene flow occurring during or following

only known remaining Indiana population.

endangered southern California

population divergence. Hence, our results

Mark-recapture surveys were conducted

mountain yellow-legged frogs,

demonstrate that unique evolutionary

from J une 20 0 8 – October 20 0 8 and J uly

Rana muscosa

lineages of R. muscosa occupy each

20 0 9 – September 20 0 9 at 35 southern

Indiana sites. Despite considerably higher Sean. D. Schoville, Tate S. Tunstall, Vance

mountain range in southern California and

T. Vredenburg, Adam R. Backlin, Elizabeth

survey efforts than previous studies, we Gallegos, Dustin A. Wood & Robert N. Fisher

should be managed separately. We also show

documented fewer total captures and ountain yellow-legged frogs,

that some genetic variation has been lost in

M muscosa, have been listed as a federally endangered distinct population

the establishment of the captive population, Rana extremely low population densities. Density

suggesting that recruitment of additional

was estimated at 0 .0 6 individuals/ 10 0

m 2 with a catch per unit effort of 0 .0 5 segment in southern California since 20 0 2.

breeding frogs may be necessary. However,

individuals/ person hour. These results Surveys indicate that populations are now

the initial stages of the captive breeding

fall well below estimates from other extinct at >99% of historical sites in the

program have been promising, with the first

reintroduction occurring this year in April. parts of the species range and highlight a San Gabriel, San Bernardino, San J acinto

significant decline from historical surveys

and Cuyamaca mountain ranges, with only in the study area. Sex ratios were male nine small populations remaining. The Tu n s ta ll, V.T. Vre d e n bu rg, A. R.

Fu ll a rticle : Sch o ville , S.D ., T.S.

biased (2.6 males: 1 female), no sub-adult small size of these populations makes them Ba cklin , E. Ga lle go s , D .A. W o o d ,

age classes were found, and only two nests

prone to unpredictable environmental were located. The population’s bias towards

a n d R.N . Fis h e r. 2 0 11. Co n s e rva tio n

events, as was recently demonstrated when older age classes combined with low

o f e vo lu tio n a ry lin e a ge s o f th e

a catastrophic wildfire, followed by flooding e n d a n ge re d m o u n ta in ye llo w -le gge d population densities will likely negate the and heavy stream erosion, impacted the fro g, R a n a m u s co s a ( Am p h ibia :

effects any natural breeding efforts would San Bernardino population. Recently, the Ra n id a e ) , in s o u th e rn Ca lifo rn ia .

have on long-term population survival.

B io lo g ic a l Co n s e r v a t io n

San Diego Zoo Institute for Conservation

Fu ll a rticle : Bu rgm e ie r, N . e t

Research established a captive breeding

a l. ( 2 0 11) . Po p u la tio n Sta tu s o f th e Ea s te rn H e llbe n d e r ( Cr y p t o b r a n ch u s a lle g a n ie n s is

a lle g a n ie n s is ) in In d ia n a . J. H e rp

4 5 ( 2 ) : 19 5 -2 0 1.

Effects of urbanization on occupancy of stream salamanders

Steven J . Price, Kristen K. Cecala, Robert A. Browne & Michael E. Dorcas

U rban development is the most common

form of land conversion in the United States. Using a before– after control-impact study design, we investigated the effects of urbanization on larval and adult stages of southern two-lined salamanders ( Eurycea cirrigera) and northern dusky salamanders ( Desmognathus fuscus). Over 5 years, we estimated changes in occupancy and probabilities of colonization and survival in

13 stream catchments after urbanization and

Adult Rana muscosa from Riverside County, California. Photo taken by Adam Backlin.

in 17 catchments that were not urbanized. We also examined effects of proportion of

urbanized area in a catchment and distance J acinto Mountains during a drought year.

program from tadpoles salvaged in the San

Population Status of the Eastern

of the salamander population to the nearest To facilitate conservation and recovery of R.

Hellbender ( Cryptobranchus

stream on probabilities of colonization and muscosa, we examined patterns of genetic

alleganiensis alleganiensis) in

survival. Before urbanization, adult and variation, historical population connectivity,

Indiana

larval stages of the two salamander species and declines in genetic variability using Nicholas G. Burgmeier, Shem D. Unger, Trent

occupied nearly all surveyed streams, with mitochondrial and microsatellite data.

M. Sutton & Rod N. Williams

occupancy estimates ranging from 1.0 to Levels of genetic variation are low in all

0 .78. Four years after urbanization mean remaining southern California populations

T status of imperiled wildlife species, occupancy of larval and adult two-lined

o better understand the conservation

and there is strong evidence of recent

salamanders had decreased from 0 .87 genetic bottlenecks. At the same time,

rigorous examinations of current and

and 0 .78 to 0 .57 and 0 .39, respectively. genetic variation among populations is

historical population densities need

Estimates of mean occupancy of larval quite high, suggesting a high degree of

to be conducted. Eastern Hellbender

alleganiensis northern dusky salamanders decreased historical isolation within and between

( Cryptobranchus

from 1.0 to 0 .57 in urban streams 4 years mountain ranges. We fit the observed data

alleganiensis) populations have drastically

after urbanization; however, adult northern to a biogeographic model and estimate that

declined throughout much of their range

dusky salamander occupancy remained these populations diverged during glacial

during the past few decades. This study

close to 1.0 in urban streams over 5 years. episodes of the Pleistocene, with little

documents the decline of the Eastern

Hellbender over the previous 25 years in the

Occupancy estimates in control streams

FrogLog Vol. 97 | July 20 11 | 57 FrogLog Vol. 97 | July 20 11 | 57

and Lithobates sylvaticus nearer to sunset over 5 years. Urbanization was associated

of some of the rarer species at risk. The

than Pseudacris crucifer, Lithobates with decreases in survival probabilities of

creation of spatial pond persistence models

catesbeianus, or Lithobates clamitans. adult and larval two-lined salamanders

allowed the identification of areas most at

Of eight species recorded during calling and decreases in colonization probabilities

risk of further pond loss. We suggest that

surveys (the preceding five plus Scaphiopus of larval dusky salamanders. Nevertheless,

local amphibian conservation efforts will

holbrookii, Hyla versicolor, and Lithobates proportion of impervious surface and

be the most effective if the focus is on the

palustris), detection probabilities of all distance to nearest stream had little effect

marshes and dune areas and on cattle-pond

preservation in the remaining grasslands. but Scaphiopus holbrookii and Lithobates on probabilities of survival and colonization.

sylvaticus varied seasonally. Peak detection Our results imply that in the evaluation of

periods ranged from 11 to 33 days and the effects of urbanization on species, such

Fu ll a rticle : Cu ra d o N ., H a rte l T.,

peak period detection probabilities ranged as amphibians, with complex life cycles,

Arn tze n J.W . ( 2 0 11) . Am p h ibia n

from 0 .0 6 for Scaphiopus holbrookii to consideration of the effects of urbanization

p o n d lo s s a s a fu n ctio n o f la n d s ca p e

0 .84 for Pseudacris crucifer. There was on both adult and larval stages is required.

ch a n ge – A ca s e s tu d y o ve r th re e

d e ca d e s in a n a gricu ltu ra l a re a

strong to moderate support for models

o f n o rth e rn Fra n ce . Bio lo gica l

with annual variation for all species except

Fu ll a rticle : Price , S.J., K.K. Ce ca la ,

Co n s e rva tio n 14 4 ( 5 ) : 16 10 – 16 18

Hyla versicolor. Detectability was affected

R.A. Bro w n e , a n d M.E. D o rca s .

more by surface water temperature than

2 0 11. Effe cts o f u rba n iza tio n o n

by air temperature, but models with both

o ccu p a n cy o f s tre a m s a la m a n d e rs .

Effects Of Temperature And

received greater support in five species.

Co n s e rva tio n Bio lo gy 2 5 : 5 4 7-5 5 5 .

Temporal Factors On Anuran

For six species, models with temperature

Detection Probabilities At

and seasonal effect received the greatest

Cape Cod National Seashore,

support, indicating that detectability is a

Amphibian pond loss as a function Massachusetts, Usa: Implications function of both temperature and day of of landscape change – A case

For Long-Term Monitoring

year. Durations of peak calling periods were

study over three decades in an

long enough and detection probabilities

agricultural area of northern France Paton & Brad C. Timm

high enough to effectively monitor six Nuno Curado, Tibor Hartel & J an W. Arntzen

Robert P. Cook, Todd A. Tupper, Peter W.C.

o evaluate the efficacy of calling surveys, of the eight species with calling surveys. gricultural reform and infrastructural

T we studied effects of temperature and Fu ll a rticle : Co o k, R.P., T. Tu p p e r,

A development are among the major

temporal factors (diel, seasonal, annual)

P.W .C. Pa to n , a n d B. Tim m . 2 0 11.

drivers of biodiversity loss and landscape

on detection probabilities for anurans

Effe cts o f te m p e ra tu re a n d te m p o ra l

homogenization worldwide. Ponds, as

of southeastern Massachusetts, USA.

fa cto rs o n a n u ra n d e te ctio n

other small stagnant water bodies, have a

We used automated recording systems

p ro ba bilitie s a t Ca p e Co d N a tio n a l

significant importance on regional-scale

(ARS) to quantify diel chronology during

Se a s h o re : Im p lica tio n s fo r Lo n g-

biodiversity; yet, they are particularly

one field season, and conducted calling

Te rm Mo n ito rin g. H e rp e to lo gica l

vulnerable to destruction and increased

surveys to investigate seasonal and annual

Co n s e rva tio n a n d Bio lo gy 6 ( 2 ) :2 5 -

39.

isolation caused by human induced

variation in calling at 10 3 wetlands over

landscape change. In this study we

six years at Cape Cod National Seashore.

investigated for a set of 199 ponds if pond

Five species detected with ARS called

persistence (n = 86, 43%) or disappearance

primarily between sunset and midnight,

(n = 113, 57%) was related to wider changes

with mean calling time of Anaxyrus fowleri

in the landscape over the period 1975– 20 0 6 in an agricultural area in north-western France, and determined to what extent pond loss affects the local amphibian species’ breeding possibilities. Landscape data was obtained from the classification of aerial photographs (1963) and two satellite images (20 0 1 and 20 0 3), complemented with field observations (1975 and 2006). Land use around the ponds was described and compared over concentric circles with five different radii in the 100–1000 m range. Overall, pond disappearance was associated with a decrease in grassland and an increase in arable fields and, in a lesser extent, urban areas around the ponds. We found that the small, man-made cattle ponds, with either natural substrate or concrete drinking troughs, were more often affected than the larger, semi-natural ponds. Since the cattle ponds are regularly used for amphibian reproduction, their massive destruction

had a strong effect of local amphibian Seasonal variation in detection probabilities of calling anurans during 5-minute calling surveys at Cape Cod National

Seashore, based on model ψ (.),γ(.),ε(.), p(julian+julian 2 ) in Appendix Table 1. SCHO = Scaphiopus holbrookii , ANFO

richness and breeding sites’ availability,

= Anaxyrus fowleri, HYVE = Hyla versicolor, PSCR = Pseudacris crucifer, LICA = Lithobates catesbeianus, LICL = L.

therewith weakening the population

clamitans , LIPA = L. palustris .

network and putting the local occurrence

58 | FrogLog Vol. 97 | July 20 11

FrogLog Vol. 97 | July 20 11 | 59

T he use of explosive compounds has resulted in contamination of terrestrial and aquatic ecosystems and is currently

a serious concern in the United States, Germany, and Canada. In the United States both 2,4,6-trinitrotoluene (TNT) and 2,4- dinitrotoluene (DNT) have been detected in vicinity of munitions facilities and waste water from TNT manufacturing plants. Little is known regarding the effects of chronic exposure of TNT and DNT in early developmental stages of North American species, such as the bullfrog. Chronic aqueous exposures were conducted using bullfrog ( Rana catesbeiana) tadpoles (8 d old) exposed to TNT, 2,4-DNT, and 2,6-DNT for 90 d. Survival, growth, gross morphology, incidence of abnormal swimming and morphological abnormalities were evaluated. Survival of tadpoles was significantly reduced at all concentrations tested. Only 2,4-DNT did not have a significant effect on body mass or SVL, but all three compounds tested had significant effects on survival. Our study suggests that long-term continuous exposure to these compounds at concentrations of 0 .25 mg/L could lead to significant changes in growth and survival of larval amphibians.

Fu ll a rticle : Effe cts o f ch ro n ic 2 ,4 ,6 ,- trin itro to lu e n e , 2 ,4 -d in itro to lu e n e ,

a n d 2 ,6 -d in itro to lu e n e e xp o s u re o n d e ve lo p in g bu llfro g ( R a n a

c a t e s b e ia n a ) ta d p o le s . 2 0 11. Eco to xico lo gy a n d En viro n m e n ta l Sa fe ty ( 74 ) : 9 2 4 – 9 2 8

Effects of three explosive compounds on developing bullfrog

( Rana catesbeiana) tadpoles

Norka E. Paden, Ernest E. Smith, J onathan D. Maul & Ronald J . Kendall

Control bullfrog tadpole after two weeks (A). Mild abdominal edema in tadpole exposed to 2,6-DNT (8 mg/L) for one week (B). Tail flexure in tadpole exposed to 1mg/L of TNT for one week (C).

Diseases and Toxicology

I nfectious diseases have emerged as important factors in the declines of many amphibian populations. Helminths,

in particular, are common parasites of frogs and evidence suggests that at least some species (e.g., Ribeiroia ondatrae, Echinostoma

trivolvis)

may have detrimental effects on the growth and survival of larval and juvenile frogs. In addition, previous studies have suggested that wetland pollution and loss of native habitats in the landscapes surrounding wetlands may influence the transmission of these parasites in frog populations; however, a complete understanding of the contributions of wetland versus landscape factors on helminth transmission is lacking. To better understand the interplay between wetland and landscape factors and to identify specific environmental factors and spatial scales associated with helminth infections, we examined the mean abundances, richness and diversity of helminth taxa in

18 populations of juvenile northern leopard frogs ( Rana pipiens) in relation to wetland and landscape variables quantified at 1 km (local) and 10 km (regional) spatial extents in Minnesota (USA). Regression analyses and variance partitioning techniques revealed that variation in the abundances of parasite taxa that are found as larvae in frogs (larval trematodes) were influenced primarily by

wetland ( R 2 adj = 29.1%) and local landscape ( R 2 adj = 28.0 %) variables, whereas the abundances of parasite taxa found as adults in frogs (adult trematodes and nematodes) were significantly associated

with regional landscape variables ( R 2 adj = 60.6%). The particular variables identified as important for larval trematodes were the concentrations of atrazine and phosphorus in wetland water (positive associations), the percentage of forest habitats in the local landscapes (positive association) and size of open water habitats in the local landscapes (negative association). Large patches of woody wetland habitats in the regional landscapes were identified as important for adult parasite taxa. Forest and woody wetland habitats in surrounding landscapes were also identified as important for maintaining parasite taxa richness and diversity. In summary, our data emphasize the influence of multiple spatial scales on helminth transmission in frogs and suggest that anthropogenic activities that have resulted in the loss of the availability and connectivity of native habitats in landscapes surrounding wetlands are associated

Effects of wetland vs. landscape variables on parasite communities

of Rana pipiens: links to anthropogenic factors

Anna M. Schotthoefer, J ason R. Rohr, Rebecca

A. Cole, Anson V. Koehler, Catherine M. J ohnson, Lucinda B. J ohnson & Val R. Beasley

C ritical developmental windows are particularly important for infections that alter host development. Here, we

test the influence of host developmental stage on the risk of mortality and the types of malformations produced in Pacific chorus frogs (Pseudacris regilla) following

exposure

to trematode ( Ribeiroia ondatrae) parasite infection.

Consistent with a critical window of vulnerability, host mortality and malformations were greatest among animals exposed during pre-limb and early limb development (15– 90 %) and decreased to <5% with progressive development. Early stage animals exhibited a higher frequency of missing limbs, whereas extra limbs and limb elements developed predominantly among tadpoles exposed after limb development was initiated. Hosts infected later in limb development were normal or exhibited only minor abnormalities.

Increases in host tolerance rather than host resistance largely explained the observed changes in pathology. Prior to host metamorphosis, parasites exhibited comparable success invading host tissue, but the amount of resulting damage differed significantly as a function of host size and developmental stage. Following metamorphosis hosts were significantly more resistant to infections.

These findings highlight the importance of critical developmental windows for infectious diseases and underscore the role of developmental changes in host tolerance in controlling this process.

Fu ll a rticle : Jo h n s o n , P. T. J., E. Ke lle rm a n n s , a n d J. Bo w e rm a n .

Critical windows of disease risk: amphibian pathology driven by developmental changes in host

resistance and tolerance

Pieter T. J . J ohnson, Esra Kellermanns & J ay Bowerman

( 2 0 11) . Critica l w in d o w s o f d is e a s e ris k: a m p h ibia n p a th o lo gy d rive n by d e ve lo p m e n ta l ch a n ge s in h o s t

re s is ta n ce a n d to le ra n ce . Fu n ctio n a l Eco lo gy 2 5 : 72 6 -73 4 .

exposed 5 days post-hatch

effect was noted across all studies, it was

through Nieuwkoop Faber Stage

considered the most sensitive indicator of

62. The levels of atrazine tested

atrazine exposure measured. The changes

included those potentially found

noted in body and organ size at 20 0 and

in puddles, vernal ponds and

400 μg/L atrazine indicated chronic

runoff soon after application

developmental exposure negatively affected

(200 and 400 μg/L) and a low

the tadpoles. Significant reductions in

level studied by a number of

fat body size could potentially lessen

other investigators (25 μg/L).

their ability to survive metamorphosis

During all exposures, mortality,

or diminish reproductive fitness as frogs

growth, metamorphosis, sex

rely on stored lipids for these processes.

ratio, fat body (a lipid storage organ) size and liver weights,

Fu ll a rticle : Za ya , R.M. e t a l ( 2 0 11)

both relative to body weight, were

Atra zin e e xp o s u re a ffe cts gro w th ,

evaluated. In certain studies,

bo d y co n d itio n a n d live r h e a lth in

feeding behavior was recorded,

Xe n o p u s la e vis ta d p o le s Aqu a tic

Metacericae of Echinostoma trivolvis infecting the kidney of a

livers and fat bodies were

To xico lo gy 10 4 ; 2 4 3 -2 5 3 .

frog. This larval trematode species was the most common parasite

histologically evaluated, liver

observed in juvenile northern leopard frogs in Minnesota and has been implicated in causing tadpole death and reduced growth in laboratory

glycogen and lipid content were

studies. Credit: Jason R. Rohr.

determined by image analysis,

Agricultural intensity in ovo affects

and

immunohistochemical

growth, metamorphic development

with declines in helminth richness and

detection of activated caspase-3

and sexual differentiation in the

abundance, but that alteration of wetland

in hepatocytes was performed. No effects

common toad ( Bufo bufo)

water quality through eutrophication or

were noted at 25 μg/L. None of the exposure

pesticide contamination may facilitate

F. Orton & E. J . Routledge the transmission of certain parasite

levels changed sex ratios nor were intersex

taxa. Reducing inputs of agrochemicals ollution was cited by the Global

gonads noted, however, no definitive

P Amphibian Assessment to be the second

histological evaluation of the gonads was

into wetlands may be particularly

important for limiting the effects of larval most important cause of amphibian decline

performed. Exposure to 400 μg/L atrazine

trematode infections on frog populations. worldwide, however, the effects of the

did not increase mortality versus controls.

Although a marginal increase in mortality

agricultural environment on amphibians

at the 200 μg/L level was noted, it was not

Fu ll a rticle : Sch o tth o e fe r e t a l. ( 2 0 11)

are not well understood. In this study,

statistically significant. At the 400 μg/L

Effe cts o f w e tla n d vs . la n d s ca p e

spawn from

Bufo bufo was taken from four

level, tadpoles were smaller than controls by

va ria ble s o n p a ra s ite co m m u n itie s o f

sites in England and Wales with varying Rana pipiens: lin ks to an thro po ge n ic 72 hours of exposure and remained smaller

intensities of arable agriculture. Spawn was

for the rest of the exposure. Appetite was

fa cto rs . Eco lo gica l Ap p lica tio n s : 2 1;

either placed in tanks containing aged tap

not decreased at any exposure level. Slowed

12 5 7-12 71.

water ( ex-situ, 5 replicates) or in cages at the

metamorphosis was noted only at 400 μg/L

Schotthoefer.Anna@mcrf.mfldclin.

native site (caged, 4 replicates). Hatching

in two of five studies. No pathological

edu

success, abnormal tadpoles, and forelimb

changes or differences in glycogen or lipid

emergence were recorded during the larval

stores were noted in the livers of exposed

stage. Individuals were also sampled at five

Atrazine exposure affects growth, tadpoles. However, in one study, livers time points during development (5-, 7-, 9-,

were significantly smaller after exposure to

body condition and liver health in

12-, 15-weeks post-hatch) and analysed for

both 200 and 400 μg/L. In another study,

Xenopus laevis tadpoles

morphological parameters. The thyroids

(TP2) and the gonads (TP3,4,5) were Renee M. Zaya, Zakariya Amini, Ashley S.

livers from 400 μg/L exposed tadpoles

Whitaker, Steven L. Kohler & Charles F. Ide also analysed histologically. With the

had higher numbers of activated caspase-3

exception of the thyroid histopathology, ix studies were performed regarding the

immunopositive cells suggesting increased

S all analysed endpoints were significantly

rates of apoptosis. Fat body size decreased

different between ex-situ individuals detected pesticide in fresh water in the US,

effects of atrazine, the most frequently

significantly after exposure to 200 and 400

reared under identical conditions from on developing

μg/L although they still contained some

Xenopus laevis tadpoles lipid and lacked any pathology. Since this the different sites. In addition, intensity of arable agriculture had a negative effect on growth and development. At one site, despite distinct rearing conditions, a high level of intersex (up to 42%) and similar sex ratios were observed in both ex-situ and caged indiviuals. Taken together, these data suggest that maternal exposure and/ or events in ovo had a much larger effect on growth, metamorphic development, and sexual differentiation in Bufo bufo than the ambient environment. This could have important implications for traditional exposure scenarios that typically begin at the larval stage. Intersex is reported for the

Exposure to 200 and 400 µg/L atrazine throughout development significantly decreased fat body size in Xenopus laevis

first time in European amphibians in situ,

tadpoles (p ≤ 0.05). Fat bodies (circled) from control (A) and 400 µg/L atrazine exposed (B) stage 62 tadpoles. Arrows:

highlighting the potential use of distinct

ovaries, K: Kidneys, Bar = 1 mm.

60 | FrogLog Vol. 97 | July 20 11

FrogLog Vol. 97 | July 20 11 | 61

B oth emerging infectious diseases and environmental contamination are known to impact amphibian survival in a

wide variety of studies. While there are clear instances of epizootics occurring in pristine environments, many species of amphibians are regularly exposed to pollutants such as pesticides. Despite this being well known, few studies have examined the influence of contaminants on disease susceptibility, and even fewer have also incorporated the role of natural stressors such as predation. This experimental study investigates the interaction of the insecticide carbaryl, dragonfly predator cue, and the emerging pathogen Ambystoma tigrinum virus (ATV) on fitness correlates and disease susceptibility in tiger salamander larvae. Four week old larvae were exposed for 22 days in a 2 (0 , 500 μg/l carbaryl) × 2 (control, predator cue water) × 2 (0, 1 × 10^4 pfu ATV) factorial designed laboratory study. In this study, predator cue strongly exacerbated disease-driven mortality. There was a clear pattern of reduced survival with the addition of stressors, with those where all three stressors were present exhibiting the worst effects: a decrease in survival from 93% in controls to 60 % with all three stressors present. For the surviving larvae, there were several sub-lethal impacts in mass, SVL, and development. Predator cue and pesticide treatments significantly reduced both SVL and mass while virus and predator treatments significantly slowed

highlight the importance of examining combined natural and introduced stressors to understand potential impacts on amphibian survival and development. Experiments focused on single stressors might be drastically underestimating actual effects. In particular, runoff from agricultural areas might contribute to the emergence of ATV in particular regions, raising concerns about the influence of pesticides on disease emergence in general.

Combined effects of virus, pesticide, and predator cue on the larval tiger salamander ( Ambystoma tigrinum)

J acob L. Kerby, Alison J . Hart & Andrew Storfer

R anaviral disease has been implicated in mortality of wild and captive amphibians. It is caused by members of

the genus Ranavirus and was recently listed as a notifiable disease by the World Organization for Animal Health. In Canada, amphibian die-offs associated with ranavirus infection have occurred in Saskatchewan, Manitoba, Ontario and New Brunswick and we recently reported the first incidence of ranaviral infection in amphibians from Quebec. The infection was found in juveniles of Lithobates pipiens captured in a privately-owned wildlife preserve in Boucherville, Quebec (45.6477°N, 73.4350 °W). DNA sequences suggested that the Quebec material was related to Frog Virus 3 (FV3). Wild caught froglets exhibited high rates of mortality in the laboratory that was accompanied by erythema, ulceration of the limbs, pupillary constriction, excessive shedding of the skin and loss of the righting reflex. The rapid onset of symptoms suggests that this form of ranaviral infection may be highly lethal to frogs that are exposed to additional stressors such as transport and capture. We also observed an unusually high rate of unilateral anophthalmia (0 .8%) in the frog population. Possible links into ocular malformations and ranaviral infections warrant future investigation, given a recent publication that associated reduced eye size in an adult of Lithobates catesbeianus with infection by FV3.

Fu ll a rticle : Pa e to w , L. J. e t a l. ( 2 0 11)

Firs t D e te ctio n o f Ra n a viru s in

Lit h o b a t e s p ip ie n s in Qu e be c.

H e rp e to l. Re vie w : 4 2 ( 2 ) ; 2 11-2 14 . ( lin d a .p a e to w @gm a il.co m )

First Detection of Ranavirus in

Lithobates pipiens in Quebec.

By Linda J . Paetow, Bruce D. Pauli, J . Daniel McLaughlin, J ulie Bidulka & David J . Marcogliese

B d prevalence rates (>2.1%) in Warren is present in salamanders at very low County, Va. We calculated Bd prevalence

at >2.1% in a multispecies survey of 20 0

Bd is widespread in aquatic Appalachian salamanders, but

prevalence rates are low.

Brian Gratwicke, Matthew Evans, Evan H. Campbell Grant, J oe Greathouse, William J . McShea, Nancy Rotzel & Rob C. Fleischer.

Fu ll a rticle :Ke rby e t a l. ( 2 0 11) Co m bin e d e ffe cts o f viru s , p e s ticid e ,

a n d p re d a to r cu e o n th e la rva l tige r s a la m a n d e r ( Am b y s t o m a t ig r in u m ) Eco h e a lth D OI: 10 .10 0 7/ s 10 3 9 3 -0 11-

0 6 8 2 -1

E ngimatic declines have come to characterize the majority of decline events documented within the

global amphibian crisis in the past 40 years. The infectious fungal pathogen, Batrachochytrium dendrobatidis (Bd), has now been implicated in many of these declines. However, since Bd was only recently described in 1999, the impact of Bd on historical amphibian declines remains unclear. We investigated the role of Bd outbreak in two cases of enigmatic decline: (1) the disappearance of 40 % of anurans from Monteverde, Costa Rica, and (2) the decline of plethodontid salamanders in Mexico and Guatemala. We first prove the reliability of a molecular technique to detect Bd in formalin-preserved specimens. We swabbed, extracted, and ran qPCR for

38 specimens that had previously been examined using histology, and found these molecular techniques to be successful in detecting Bd in specimens with 83-90 % accuracy in comparison to histological results. Using this molecular technique, we sampled over 10 0 0 specimens collected from Mexico, Guatemala, and Costa Rica from the 1960 s to 20 0 0 s. We found a strong pattern of the absence of Bd over multiple years, species, and localities followed by

Bd emergence coincident with decline at all localities. Furthermore, our data supports the hypothesis that Bd is a novel

Museum specimens link the emergence of the chytrid fungus, Batrachochytrium dendrobatidis,

with the decline of neotropical amphibians

Tina L. Cheng, Sean M. Rovito, David B. Wake & Vance T. Vredenburg

populations of amphibians in fundamental research into the aetiology of specific developmental effects in wild amphibians.

Fu ll a rticle : Orto n , F. & Ro u tle d ge ,

E. ( 2 0 11) Agricu ltu ra l in te n s ity in o vo a ffe cts gro w th , m e ta m o rp h ic

d e ve lo p m e n t a n d s e xu a l

d iffe re n tia tio n in th e co m m o n to a d ( Bu fo b u fo ) . Eco to xico lo gy: 2 0 ;

9 0 1-9 11.

terrestrial and aquatic salamanders. Bd was detected only on Desmognathus monticola. Other studies testing for Bd on salamanders in the Appalachians found very low occurrences and the actual level of threat that Bd poses to wild Appalachian salamander populations remains an unresolved question. Our observations in Virginia are consistent with observations elsewhere in the Appalachian region, where others have found that though Bd appears to be widely distributed it is found at low prevalence rates and is predominantly found

Fu ll a rticle : B Gra tw icke , M Eva n s , E H Ca m p be ll Gra n t, J Gre a th o u s e , W J McSh e a , N Ro tze l, RC Fle is ch e r. Lo w p re va le n ce o f

B a t r a c h o c h y t r iu m d e n d r o b a t id is

d e te cte d in a qu a tic Ap p a la ch ia n s a la m a n d e rs fro m W a rre n Co u n ty, Virgin ia , U SA. H e rp e to lo gica l Re vie w , 2 0 11, 4 2 ( 2 ) , 2 17– 2 19 .

62 | FrogLog Vol. 97 | July 20 11

nitrate fertilizer and predator stress on larval Western Spadefoot toad ( Pelobates cultripes) from two nearby locations from Western Spain, using adult caged male marbled newts (Triturus m arm oratus) as predators. Local differences were detected in terms of larval growth, which was affected by the combination of stressors in one of the two populations tested. Whereas tadpoles exposed to 45.2 mg N-NH4+/ L were 7 % smaller than controls, the presence of predators from a foreign community resulted in animals 15 % larger than without predators. Interestingly, predators from the same community as the tadpoles did not affect larval growth. The length of the tadpoles from a nearby locality was unaffected after exposure to

spreading pathogen. Our results show that Bd emerged in Mexico in the early 1970 s and subsequently spread south to Guatemala by the 1980s/1990s and finally to Monteverde, Costa Rica by 1987. We also show lethality in two species of plethodontid salamanders ( Bolitoglossa rufescens, Pseudoeurycea leprosa) and resistance in a Mexican frog, Plectrohyla matudai, which could potentially be one of the many carrier hosts spreading Bd from aquatic habitats to terrestrial environments. This study highlights the potential use of museum specimens to answer questions regarding

Bd emergence and spread in the scope of amphibian decline, and further contributes to the complex ecology of Bd by describing its impact in a group of terrestrial amphibians, plethodontid salamanders.

Fu ll a rticle : Ch e n g TL, Ro vito SM, W a ke D B, Vre d e n bu rg VT ( 2 0 11) Co in cid e n t m a s s e xtirp a tio n o f n e o tro p ica l a m p h ibia n s w ith th e e m e rge n ce o f th e in fe ctio u s p a th o ge n , Ba t r a ch o ch y t r iu m

Spatial-temporal spread of Bd found from museum sampling, including “ Bd -wave” from Lips et al. (2008). Map by Sean M. Rovito

T adpoles use chemical signals to detect predators such as dragonfly larvae and respond by changing their body and

tail shape. The new phenotype gives them greater protection from predation. There is concern that pollutants such as metals and pesticides may affect the ability of tadpoles to respond to the naturally occurring chemical signals from predators. Elevated concentrations of metals such as copper and zinc are often found in natural ecosystems. The aim of this study was to measure the effects of increasing concentrations of these

metals on the response of tadpoles to chemical signals from dragonfly larvae. The study used tadpoles of the Arabian toad

( Bufo

arabicus) and exposed them to low levels of copper and zinc in water that had previously contained dragonfly larvae ( Crocothemis

erythrea) that were fed tadpoles, or water that had contained only tadpoles. Tadpoles that received the predator- water, developed deeper bodies and slightly longer tail fins than controls. But when tadpoles were grown in predator-water plus 35 μg/L copper they reverted to close to the control phenotype. The copper appears to have either inhibited the development of the protective phenotype or has prevented the tadpoles from detecting the dragonfly smell. On

the other hand zinc did not have any effect on the tadpoles at the low concentrations used in this study. The study shows

Effects of copper, zinc and dragonfly kairomone on growth rate and

induced morphology of Bufo arabicus tadpoles

Michael J Barry

A griculture-related pollution is undoubtedly a major risk for many amphibian populations, but even within

a small area, diffuse agricultural pollution does not affect all aquatic environments equally, which could account for local differences in amphibians’ sensitivity to agrochemicals. Furthermore, ecological stressors to which amphibians are exposed in the field, such as predation pressure, can make agrochemicals far more deadly that when acting in isolation. We examined the combined effects of ammonium

Responses of toad tadpoles to ammonium nitrate fertilizer and

predatory stress: Differences between populations on a local scale

Manuel E. Ortiz-Santaliestra, María J . Fernández-Benéitez, Miguel Lizana & Adolfo Marco

Snout-vent length (SVL) of larvae from two Pelobates cultripes populations exposed to ammonium nitrate and predatory marbled newts. Lowercase letters indicate homogeneous groups defined by Tukey post-hoc tests.

ammonium nitrate and predatory stress.

Fu ll a rticle : Ortiz-Sa n ta lie s tra , M.E. e t a l. ( 2 0 11) Re s p o n s e s o f to a d ta d p o le s to a m m o n iu m n itra te

fe rtilize r a n d p re d a to ry s tre s s :

D iffe re n ce s be tw e e n p o p u la tio n s o n a lo ca l s ca le . En viro n m e n ta l To xico lo gy a n d Ch e m is try 3 0 : 14 4 0 -

14 4 6 .

FrogLog Vol. 97 | July 20 11 | 63

C hytrid fungus Batrachochytrium dendrobatidis (Bd) was first detected

in Britain in 20 0 4 and since that time has achieved high prevalence in many natterjack toad ( Bufo calamita) populations, indeed consistently higher than in any other British amphibian species so far. We quantified aspects of larval fitness, adaptive (major histocompatibility

complex)

diversity

and neutral (microsatellite) diversity in natterjack populations in two regions of north-west England. Toads in region one had no evidence of chytrid infection whereas in region two there was a substantial prevalence of Bd. Larval fitness (growth rate, time to metamorphosis and survival) of

B. calamita did not differ between the regions. Genetic diversity at microsatellite loci was much higher in the infected than in the uninfected region but the converse was true of MHC diversity, indicating that genetic drift was unlikely to explain the differences in MHC between the regions. Furthermore, MHC allele frequencies varied significantly between Bd- infected and uninfected populations. Microsatellite diversity was not a robust indicator of larval fitness in these toad populations while MHC genotype frequencies varied in

a way that was consistent with directional selection in response to pathogen prevalence.

The acquired

immune

defences may therefore play an important role in determining the susceptibility of amphibians to chytridiomycosis although this requires much wider investigation both in other natterjack populations and in a range of other species. Thus far there is no evidence of decline in any of the infected natterjack toad populations, providing some hope that this species might prove resistant to the pathogen.

Fu ll a rticle : Co n s e rva tio n Ge n e tics

12 , 8 0 5 -8 11 ( 2 0 12 )

Larval fitness and immunogenetic diversity in chytrid-infected and uninfected natterjack toad ( Bufo calamita) populations

Shoshanna May, Inga Zeisset & Trevor J .C. Beebee

that metallic pollutants such as copper may affect indirect interactions between species in ways that we did not previously suspect.

Fu ll a rticle : Ba rry, M.J. ( 2 0 11) Effects of copper, zinc and dragonfly ka iro m o n e o n gro w th ra te a n d

in d u ce d m o rp h o lo gy o f Bu fo

a r a b ic u s ta d p o le s . Eco to xico lo gy

a n d En viro n m e n ta l Sa fe ty 74 :4 9 18 - 923

Consequences of an amphibian malformity for development and

fitness in complex environments

Matt J . Michel & Stephanie Burke

(a) Non- scoliotic and (b) scoliotic tadpoles. Photo: Matt J. Michel