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No detection of the pathogen Batrachochytrium dendrobatidis in Sardinian cave salamanders, genus Hydromantes

Identifieur interne : 000E03 ( Istex/Corpus ); précédent : 000E02; suivant : 000E04

No detection of the pathogen Batrachochytrium dendrobatidis in Sardinian cave salamanders, genus Hydromantes

Auteurs : Ylenia Chiari ; Arie Van Der Meijden ; Mauro Mucedda ; Norman Wagner ; Michael Veith

Source :

RBID : ISTEX:E27F0CD359E3F82A8C57EC50DEC957D253D0EB64

Abstract

Amphibian declines have been documented worldwide. Chytridiomycosis, a disease caused by the aquatic fungus Batrachochytrium dendrobatidis (Bd), is one of the causes associated with these declines. Occurrence, rate of infection and mortality due to chytridiomycosis in amphibians depend on multiple factors including habitat and life-style (aquatic/terrestrial). Bd infection is lower in terrestrial than in aquatic species, but a fully terrestrial life-style alone may not explain the absence of Bd in some species. Low individual dispersal, decreasing the occurrence of contact with infected organisms, could also favour lower Bd infection. To survey the occurrence of Bd infection in fully terrestrial salamanders with low dispersal, we sampled the Sardinian Hydromantes species to measure their level of infection. Bd was not detected and likely absent in Sardinian Hydromantes. This phenomenon could be explained by a combination of terrestrial habitat, low dispersal, and occurrence mostly in habitats where other amphibians do not occur.

Url:
DOI: 10.1163/15685381-00002876

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ISTEX:E27F0CD359E3F82A8C57EC50DEC957D253D0EB64

Le document en format XML

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<idno type="eISSN">1568-5381</idno>
<imprint>
<publisher>Brill</publisher>
<pubPlace>Netherlands</pubPlace>
<date type="published" when="2013"></date>
<biblScope unit="volume">34</biblScope>
<biblScope unit="issue">1</biblScope>
<biblScope unit="page" from="136">136</biblScope>
<biblScope unit="page" to="141">141</biblScope>
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<idno type="istex">E27F0CD359E3F82A8C57EC50DEC957D253D0EB64</idno>
<idno type="DOI">10.1163/15685381-00002876</idno>
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<date>2013</date>
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<abstract>
<p>Amphibian declines have been documented worldwide. Chytridiomycosis, a disease caused by the aquatic fungus Batrachochytrium dendrobatidis (Bd), is one of the causes associated with these declines. Occurrence, rate of infection and mortality due to chytridiomycosis in amphibians depend on multiple factors including habitat and life-style (aquatic/terrestrial). Bd infection is lower in terrestrial than in aquatic species, but a fully terrestrial life-style alone may not explain the absence of Bd in some species. Low individual dispersal, decreasing the occurrence of contact with infected organisms, could also favour lower Bd infection. To survey the occurrence of Bd infection in fully terrestrial salamanders with low dispersal, we sampled the Sardinian Hydromantes species to measure their level of infection. Bd was not detected and likely absent in Sardinian Hydromantes. This phenomenon could be explained by a combination of terrestrial habitat, low dispersal, and occurrence mostly in habitats where other amphibians do not occur.</p>
</abstract>
<textClass>
<keywords scheme="keyword">
<list>
<item>
<term>Short Notes</term>
</item>
</list>
</keywords>
</textClass>
<textClass>
<keywords scheme="keyword">
<list>
<head>keywords</head>
<item>
<term>amphibian decline</term>
</item>
<item>
<term>amphibians</term>
</item>
<item>
<term>Batrachochytrium dendrobatidis</term>
</item>
<item>
<term>low dispersal</term>
</item>
<item>
<term>pathogen infection</term>
</item>
<item>
<term>terrestrial salamander</term>
</item>
</list>
</keywords>
</textClass>
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<change when="2013">Created</change>
<change when="2013">Published</change>
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<istex:metadataXml wicri:clean="corpus brill-journals not found" wicri:toSee="no header">
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<article article-type="other">
<front>
<journal-meta>
<journal-id journal-id-type="e-issn">15685381</journal-id>
<journal-title>Amphibia-Reptilia</journal-title>
<abbrev-journal-title>AMRE</abbrev-journal-title>
<issn pub-type="ppub">0173-5373</issn>
<issn pub-type="epub">1568-5381</issn>
<publisher>
<publisher-name>Brill</publisher-name>
<publisher-loc>Netherlands</publisher-loc>
</publisher>
</journal-meta>
<article-meta>
<article-id pub-id-type="doi">10.1163/15685381-00002876</article-id>
<article-categories>
<subj-group>
<subject>Short Notes</subject>
</subj-group>
</article-categories>
<title-group>
<article-title>No detection of the pathogen
<italic>Batrachochytrium dendrobatidis</italic>
in Sardinian cave salamanders, genus
<italic>Hydromantes</italic>
</article-title>
</title-group>
<contrib-group>
<contrib contrib-type="author">
<name>
<surname>Chiari</surname>
<given-names>Ylenia</given-names>
</name>
<xref ref-type="aff" rid="aff001"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>van der Meijden</surname>
<given-names>Arie</given-names>
</name>
<xref ref-type="aff" rid="aff001"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Mucedda</surname>
<given-names>Mauro</given-names>
</name>
<xref ref-type="aff" rid="aff002"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Wagner</surname>
<given-names>Norman</given-names>
</name>
<xref ref-type="aff" rid="aff003"></xref>
</contrib>
<contrib contrib-type="author">
<name>
<surname>Veith</surname>
<given-names>Michael</given-names>
</name>
<xref ref-type="aff" rid="aff003"></xref>
<xref ref-type="author-notes" rid="fn001">*</xref>
</contrib>
</contrib-group>
<aff id="aff001">
<sup>1</sup>
Centro de Investigação em Biodiversidade e Recursos Genéticos, Universidade do Porto, Campus Agrário de Vairão, Vairão, Portugal</aff>
<aff id="aff002">
<sup>2</sup>
Gruppo Speleologico Sassarese, Sassari, Italy</aff>
<aff id="aff003">
<sup>3</sup>
Biogeography Department, Trier University, 54286 Trier, Germany</aff>
<author-notes>
<fn id="fn001">
<p>
<sup>*</sup>
Corresponding author; e-mail:
<email>veith@uni-trier.de</email>
</p>
</fn>
</author-notes>
<pub-date pub-type="ppub">
<year>2013</year>
</pub-date>
<pub-date pub-type="epub">
<year>2013</year>
</pub-date>
<volume>34</volume>
<issue>1</issue>
<fpage>136</fpage>
<lpage>141</lpage>
<history>
<date>
<day>15</day>
<month>10</month>
<year>2012</year>
</date>
<date date-type="received">
<day>22</day>
<month>01</month>
<year>2013</year>
</date>
<date date-type="accepted">
<day>22</day>
<month>01</month>
<year>2013</year>
</date>
</history>
<permissions>
<copyright-statement>© Koninklijke Brill NV, Leiden, The Netherlands</copyright-statement>
<copyright-year>2013</copyright-year>
</permissions>
<self-uri content-type="pdf" xlink:href="15685381_034_01_S16_text.pdf"></self-uri>
<abstract>
<p>Amphibian declines have been documented worldwide. Chytridiomycosis, a disease caused by the aquatic fungus
<italic>Batrachochytrium dendrobatidis</italic>
(
<italic>Bd</italic>
), is one of the causes associated with these declines. Occurrence, rate of infection and mortality due to chytridiomycosis in amphibians depend on multiple factors including habitat and life-style (aquatic/terrestrial).
<italic>Bd</italic>
infection is lower in terrestrial than in aquatic species, but a fully terrestrial life-style alone may not explain the absence of
<italic>Bd</italic>
in some species. Low individual dispersal, decreasing the occurrence of contact with infected organisms, could also favour lower
<italic>Bd</italic>
infection. To survey the occurrence of
<italic>Bd</italic>
infection in fully terrestrial salamanders with low dispersal, we sampled the Sardinian
<italic>Hydromantes</italic>
species to measure their level of infection.
<italic>Bd</italic>
was not detected and likely absent in Sardinian
<italic>Hydromantes</italic>
. This phenomenon could be explained by a combination of terrestrial habitat, low dispersal, and occurrence mostly in habitats where other amphibians do not occur.</p>
</abstract>
<kwd-group>
<kwd>amphibian decline</kwd>
<kwd>amphibians</kwd>
<kwd>
<italic>Batrachochytrium dendrobatidis</italic>
</kwd>
<kwd>low dispersal</kwd>
<kwd>pathogen infection</kwd>
<kwd>terrestrial salamander</kwd>
</kwd-group>
</article-meta>
</front>
<body>
<p>Global climate change has dramatically facilitated the worldwide spread of pathogens (Fisher et al., 2012). Among these, the aquatic chytridiomycete fungus
<italic>Batrachochytrium dendrobatidis</italic>
(
<italic>Bd</italic>
) is regarded as a major threat to amphibians worldwide (Fisher, Garner and Walker, 2009), with one expansive virulent lineage considered to be responsible for a decline of amphibian populations on large geographic scales (Farrer et al., 2011). This fungus causes chytridiomycosis, a lethal skin disease of amphibians. The amphibian skin is physiologically active, regulating the exchange of respiratory gases, water, and electrolytes (Hillman et al., 2009). It has been shown that
<italic>Bd</italic>
infection causes mortality in amphibians by disrupting normal regulatory functions of the amphibian skin and causing electrolyte depletion and osmotic imbalance (Berger et al., 1998; Voyles et al., 2007, 2009; see also Rosenblum et al., 2010 for a review).</p>
<p>In principle,
<italic>Bd</italic>
spreads through aquatic zoospores (Rosenblum et al., 2010). Therefore, an important life history trait that affects
<italic>Bd</italic>
susceptibility may be habitat type (e.g., Lips et al., 2006).
<italic>Bd</italic>
occurrence in fully terrestrial amphibians is generally much lower than in aquatic or semi-aquatic species (e.g. Kriger and Hero, 2007), but it can be found in some terrestrial species, especially if they have some aquatic life-stages during their development (e.g., Cummer, Green and O’Neill, 2005; Weinstein, 2009).</p>
<p>In Europe,
<italic>Bd</italic>
infects many but not all amphibian species and it cannot be easily predicted which species will be affected (Garner et al., 2005; Sztatecsny and Glaser, 2011). Lötters et al. (2012) recently showed that throughout its distribution range the fully terrestrial and viviparous Alpine salamander,
<italic>Salamandra atra</italic>
was not infected with
<italic>Bd</italic>
, although in the Swiss and Austrian Alps other amphibian species with an aquatic larval stage were at least in parts infected (Sztatecsny and Glaser, 2011; Tobler, Borgula and Schmidt, 2012). Lötters et al. (2012) proposed that, among other factors, a purely terrestrial mode of life could also contribute to the absence of
<italic>Bd</italic>
in
<italic>S. atra</italic>
(see Bielby et al., 2008 for a general discussion of the potential link between life history traits and infection occurrence in amphibians). However, a phylogenetically closely related and only partly terrestrial species,
<italic>S. salamandra</italic>
(fully terrestrial in its adult stage), is a European amphibian species that at least regionally suffers from population declines due to chytridiomycosis (Bosch and Martínez-Solano, 2006). This would weaken the hypothesis that a partly terrestrial habitat alone, at least during some of the developmental stages, could prevent
<italic>Bd</italic>
infection in European salamanders. However, a fully terrestrial habitat could be related to a lower occurrence of
<italic>Bd</italic>
infection in these animals. Another factor that could contribute to limit
<italic>Bd</italic>
infection could also be a limited individual dispersal, as this would decrease the contact probability with infected amphibian species which could transmit the pathogen. To this extent, we should expect that fully terrestrial species with limited individual dispersal such as those belonging to the genus
<italic>Hydromantes</italic>
(Plethodontidae) in Sardinia (Chiari et al., 2012) should show low levels of
<italic>Bd</italic>
infection. Furthermore, specific habitat requirements characteristic of European
<italic>Hydromantes</italic>
 – the animals occur mostly either in caves or deeply burrowed under layer of stones (see Van der Meijden et al., 2009 and Chiari et al., 2012 for further information) – may also limit contact with other potentially infected amphibian species and water sources therefore contributing to a low rate of infections in these animals. Therefore, we expect that in support of the life history and low dispersal hypothesis, all
<italic>Hydromantes</italic>
specimens would be protected from
<italic>Bd</italic>
exposure and free of
<italic>Bd</italic>
or have low level of
<italic>Bd</italic>
infection. To assess the validity of this suggestion, we sampled
<italic>Hydromantes</italic>
across all Sardinian species and areas. All eight species of European cave salamanders, genus
<italic>Hydromantes</italic>
, are fully terrestrial during their life cycle, with seven being oviparous and direct developing and one,
<italic>H. sarrabusensis</italic>
, hypothesized to be viviparous (Lanza et al., 2005). On the Italian island of Sardinia, where five
<italic>Hydromantes</italic>
species are endemic,
<italic>Bd</italic>
has been detected in several species, such as
<italic>Euproctus platycephalus</italic>
, a mostly aquatic salamander (although it can also seldom occur on land), and
<italic>Discoglossus sardus</italic>
, an anuran species that may be found in both aquatic and terrestrial environments (Bovero et al., 2008; Bielby et al., 2009;
<uri>http://www.bd-maps.net/maps/</uri>
, inquiry 29.08.2012).</p>
<p>
<italic>Bd</italic>
infection is temperature-dependent, showing seasonal variation. For this reason, sampling was carried out during spring, when temperature should be optimal for
<italic>Bd</italic>
occurrence (e.g., Lannoo et al., 2011). In spring 2008, we collected 143
<italic>Hydromantes</italic>
specimens from 17 Sardinian populations (table 1, fig. 1).
<italic>Hydromantes</italic>
can be difficult to find in its terrestrial habitats, and this holds true especially for cave populations. Therefore, the average sample size per population and species is 8.4 and 23.8 respectively, which is not necessarily sufficient to reliably infer absence of the pathogen (required sample size of minimum 30 individuals, DiGiacomo and Koepsell, 1986). Sampling was carried out by swabbing with ordinary cotton cosmetic swabs the ventral surfaces of body as well as fore and hind legs of salamanders. Swabs were kept at ambient temperature (which was never above 20°C) during field work and frozen at −80°C upon return from the field (Hyatt et al., 2007).</p>
<table-wrap>
<label>
<bold>Table 1.</bold>
</label>
<caption>
<p>Sardinian
<italic>Hydromantes</italic>
populations tested for
<italic>Bd</italic>
occurrence. Last column indicates the Bayesian 95% credible intervals.</p>
</caption>
<graphic xlink:href="15685381_034_01_S16_i0001.jpg" alt-version="yes"></graphic>
</table-wrap>
<fig>
<label>
<bold>Figure 1.</bold>
</label>
<caption>
<p>Sampling sites of
<italic>Hydromantes</italic>
in Sardinia, Italy. Localities 5-6, 8-9:
<italic>H. flavus</italic>
; localities 12-13, 20-26:
<italic>H. imperialis</italic>
; localities 34, 36:
<italic>H. supramontis</italic>
; locality 38:
<italic>H. sarrabusensis</italic>
; localities 43, 45:
<italic>H. genei</italic>
. Modified from Chiari et al. (2012).</p>
</caption>
<graphic xlink:href="15685381_034_01_S16_i0002.png" alt-version="yes"></graphic>
</fig>
<p>For
<italic>Bd</italic>
screening, we used quantitative real-time PCR (polymerase chain reaction) of the ITS-1/5.8S ribosomal DNA region of
<italic>Bd</italic>
following the protocol of Boyle et al. (2004), but running a duplicate for each sample. We used a
<italic>Bd</italic>
standard from ecogenics (Zurich, Swizerland) of concentrations 0.1, 1.0, 10.0 and 100.0 according to Hyatt et al. (2007). For each run, we used a positive control (sample with a known high
<italic>Bd</italic>
concentration) to rule out that PCR inhibition may have caused negative results. To obtain a Bayesian 95% credible interval for prevalence, we used the R2WinBUGS package and WinBUGS to estimate the posterior distribution of prevalence (Kéry, 2010; Lötters et al., 2012). We used a uniform prior for prevalence (e.g., prevalence ∼ U(0,1)). Three parallel Markov chains with 20 000 iterations each were run discarding the first 5000 iterations as burn-in. Chains were not thinned.</p>
<p>Our results indicate that all the samples of Sardinian
<italic>Hydromantes</italic>
tested in this study were
<italic>Bd</italic>
negative (table 1) and therefore most likely not infected. However, due to the small sample size of some populations, the 95% credible intervals are very wide and therefore reliable detection of
<italic>Bd</italic>
absence may be more uncertain (see table 1). Posterior distributions were left-skewed towards zero and all 95% credible intervals included a prevalence of zero. In other plethodontid salamanders, such as the ones studied here,
<italic>Bd</italic>
infection has been observed in wild-captured animals (e.g., the American salamander
<italic>Bolitoglossa dofleini</italic>
, Pasmans, Zwart and Hyatt, 2004), and infected animals have been shown to die under experimental conditions (e.g., in
<italic>Plethodon metcalfi</italic>
, Vazquez, Rothermel and Passier, 2009). While
<italic>Bd</italic>
is an aquatic fungus and aquatic amphibian species are more susceptible to
<italic>Bd</italic>
infection, Vazquez, Rothermel and Passier (2009) suggest that as other Chytridiomycetes,
<italic>Bd</italic>
may survive in moist environments and may be spread among amphibian species with different habitat requirements (e.g., aquatic versus terrestrial). Wake and Vredenburg (2008) proposed the possible influence of the vagile and terrestrial frog
<italic>Pseudacris regilla</italic>
on which
<italic>Bd</italic>
has been detected in explaining the wave of die-offs due to chytridiomycosis observed in the yellow-legged frogs. Low individual dispersal as shown in Sardinian
<italic>Hydromantes</italic>
(at least according to mitochondrial data, Chiari et al., 2012) could limit the transmission of
<italic>Bd</italic>
from other species. We suggest that the ecology of Sardinian
<italic>Hydromantes</italic>
at least in part may explain the absence of
<italic>Bd</italic>
in these species. First, the animals are terrestrial and
<italic>Bd</italic>
is an aquatic fungus. Second, these salamanders mostly occur in a different habitat from the one of other amphibian species on Sardinia where
<italic>Bd</italic>
and chytridiomycosis were found. Third, the low dispersal proposed for Sardinian
<italic>Hydromantes</italic>
could reduce the contact with other, potentially
<italic>Bd</italic>
-infected species. However, it has been shown that susceptibility for the pathogen may in part be host-specific, either due to the kind and composition of skin peptides, microbial communities, or due to host-specific genetic and life history traits (e.g. Harris et al., 2006; Woodhams et al., 2007a, b; Becker and Harris, 2010; Savage and Zamudio, 2011; Searle et al., 2011). We cannot exclude that our findings may be associated with skin characteristics and skin bacterial communities occurring in these species, as for example already observed and experimentally tested in another plethodontid salamander (
<italic>Plethodon cinereus</italic>
, Becker and Harris, 2010). Further studies should assess potential
<italic>Bd</italic>
pathogen-repellent skin characteristics of Sardinian
<italic>Hydromantes</italic>
. Future researches on
<italic>Bd</italic>
infection, distribution and spread should focus on understanding the influence of amphibian community composition and amphibian dispersal on infection rates and transmission of the
<italic>Bd</italic>
pathogen to fully terrestrial species.</p>
</body>
<back>
<ack>
<title>
<bold>Acknowledgements</bold>
</title>
<p>We are indebted to the speleologists of the Gruppo Grotte Ogliastra and to Maria Luisa Bertelli, Gesuino Chighini, Sandro Corona, Luciano Cuccu, Paolo Marcia, Franco Meloni, Maria Adelaide Oggiano, Carlo Onnis, Ermanno Pidinchedda, Marco Pisano, Titino Salis, Laura Sanna, Giuseppe Sotgiu, Renzo Tocco, and “il fontaniere” of Tertenia for their help with the sampling. Linda Harant and Karin Fischer helped with the laboratory work. We are grateful to Stefan Lötters, Benedikt Schmidt, and two anonymous reviewers for their comments on this article. We are thankful to the Ministero dell’Ambiente, della Tutela del Territorio e del Mare for issuing the collecting permit 9333 (03/04/2007). YC and AvdM were financially supported by FCT, Fundação para a Ciência e Tecnologia (Portugal), postdoctoral fellowships SFRH/BDP/73515/2010 and SFRH/BDP/48042/2008, respectively.</p>
</ack>
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<namePart type="given">Ylenia</namePart>
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<abstract>Amphibian declines have been documented worldwide. Chytridiomycosis, a disease caused by the aquatic fungus Batrachochytrium dendrobatidis (Bd), is one of the causes associated with these declines. Occurrence, rate of infection and mortality due to chytridiomycosis in amphibians depend on multiple factors including habitat and life-style (aquatic/terrestrial). Bd infection is lower in terrestrial than in aquatic species, but a fully terrestrial life-style alone may not explain the absence of Bd in some species. Low individual dispersal, decreasing the occurrence of contact with infected organisms, could also favour lower Bd infection. To survey the occurrence of Bd infection in fully terrestrial salamanders with low dispersal, we sampled the Sardinian Hydromantes species to measure their level of infection. Bd was not detected and likely absent in Sardinian Hydromantes. This phenomenon could be explained by a combination of terrestrial habitat, low dispersal, and occurrence mostly in habitats where other amphibians do not occur.</abstract>
<note type="footnotes">*Corresponding author; e-mail: veith@uni-trier.de</note>
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