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1

Good DNA from bat droppings

100%

Puechmaille S. J. , Mathy G. , Petit E. J.

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tom 09

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nr 1

EN

Amplification of a mitochondrial DNA fragment was used to compare the efficiency of five methods for extracting DNA from bat droppings. The Qiagen DNA Stool Kit, which yielded > 90% mtDNA amplification success, was chosen to extract DNA from 586 samples taken over two years in three French colonies of the lesser horseshoe bat (Rhinolophus hipposideros). Samples, for which mtDNA amplification was successful, were subject to the multiplex amplification of eight microsatellite loci. This resulted in > 95% amplification success over 12,592 PCRs. Allelic dropout (ADO) and false allele (FA) rates were low, and consequently, sample and locus quality indexes (QI) were high. These results demonstrate that large scale noninvasive studies of bat colonies are possible.

2

Horseshoe bats recognise the sex of conspecifics from their echolocation calls

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Schuchmann M. , Puechmaille S. J. , Siemers B. M.

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nr 1

EN

Echolocation calls produced by bats contain information on the emitter that could be used in social interactions. Nevertheless, little is known about the whether such information actually is being used by bats. Here we tested the ability of two horseshoe bat species (Rhinolophus mehelyi and R. euryale; n = 20 each) to discriminate conspecific male from conspecific female echolocation calls and vice versa. Behavioural data from a habituation-discrimination paradigm revealed that both species were able to recognize the sex of conspecifics from their calls, providing evidence that cues on the sex of the emitter are encoded in echolocation signals. While our data support that the sex-specific acoustic cues are not (just) the horseshoe bats' constant-frequency call part, their nature remains enigmatic. This is the second study to show that bats can recognize the caller's gender from echolocation calls and the first to do so for ‘high-duty-cycle bats’; those that use calls with a prominent constant-frequency (CF) component.

3

Effect of sample preservation methods on the viability of Geomyces destructans, the fungus associated with white-nose syndrome in bats

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Puechmaille S. J. , Fuller H. , Teeling E. C.

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tom 13

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nr 1

EN

Geomyces destructans is a fungus directly associated with white-nose syndrome (WNS), a recently discovered disease that has caused the death of over one million bats in North America and therefore has prompted intense scientific investigation into its biology. If precautions are not taken, the spread of G. destructans by human transportation for scientific study may be a substantial threat to bat populations. We investigated the viability of G. destructans after being exposed to commonly used DNA/RNA preservation methods. Our first experiment revealed that G. destructans is able to germinate after an eight day storage period in RNAlater or kept dry but could not germinate after storage in 70% or absolute ethanol for the same time period. Storing G. destructans samples at different temperatures did not substantially affect the results. In a second experiment, we showed that G. destructans conidia were only killed after being stored in 70% ethanol for a minimum of 24 hours while only 30 minutes were necessary when stored in absolute ethanol. Our results suggest that the DNA/RNA preservation method has an important impact on the ability of G. destructans to remain viable and should therefore be considered before samples are transported, especially to regions where the fungus has not yet been documented. Our results also strongly advocate the use of high concentrations (i.e., absolute) ethanol over 70% ethanol to rapidly kill G. destructans.

4

Towards navigating the Minotaur's labyrinth: cryptic diversity and taxonomic revision within the speciose genus Hipposideros (Hipposideridae)

88%

Foley N. M. , Goodman S. M. , Whelan C. V. , Puechmaille S. J. , Teeling E.

Acta Chiropterologica

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2017

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tom 19

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nr 1

EN

Recent molecular evidence has shown that the largest genus of the family Hipposideridae, Hipposideros, is paraphyletic with respect to H. commersonii sensu lato and H. vittatus, both belonging to a species complex referred to as the commersonii group. The taxonomic issues at the generic level of certain species of Hipposideros remain unresolved in part related to insufficient material in previous molecular studies. Herein, we expand sampling of the commersonii group and include H. commersonii sensu stricto from its type locality, Madagascar. Our phylogenetic analysis revealed that the commersonii group forms a highly supported monophyletic clade with H. cyclops, which is sister taxa to Aselliscus and Coelops. A combination of phylogenetic and comparative morphological analyses, as well as divergence time estimates, were used to provide compelling evidence to support the placement of the clade containing the commersonii group and that with H. cyclops in two resurrected genera, Macronycteris and Doryrhina, respectively. Divergence time estimates indicated that Macronycteris and Doryrhina diverged 19 mya and separated from Coelops and Aselliscus in the Oligocene, about 31 mya. The commersonii group underwent a rapid radiation as recently as 3 mya likely in response to favourable climatic conditions during the Late Pliocene in Africa. Phylogenetic analysis of Cyt-b could not resolve relationships within this morphologically conserved complex. Further sampling is necessary to fully elucidate the evolutionary history of Doryrhina. Given that cryptic species are widespread among bats, including within the genus Hipposideros, this study highlights the shortcomings of current chiropteran taxonomy to describe hidden diversity.

5

The effects of human-mediated habitat fragmentation on a sedentary woodland-associated species (Rhinolophus hipposideros) at its range margin

88%

Dool S. E. , Puechmaille S. J. , Kelleher C. , McAney K. , Teeling E. C.

Acta Chiropterologica

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2016

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tom 18

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nr 2

EN

Among the many anthropogenic modifications to earth's ecosystems, habitat loss and degradation pose the most immediate threat to many biota. The predicted consequences of fragmented habitats include lower species diversity, smaller population sizes, disrupted gene flow, increased drift and inbreeding and increased differentiation between neighbouring populations; all of which are thought to be further enhanced in species with low dispersal abilities. These factors, especially when occurring in tandem, can lead to an increased risk of extinction. To examine the genetic consequences of habitat fragmentation we selected an isolated population of a sedentary woodland specialist species (Rhinolophus hipposideros) to act as an indicator of disruptions to landscape level connectivity. Based on 491 individuals from 37 colonies our results revealed the presence of a broad North-Range/South-Range differentiation within this species in Ireland; a finding supported across datasets (mtDNA and nuclear microsatellites) and analyses. Analyses of echolocation data and microsatellites suggested further differentiation of the northern-most colonies. A landscape genetics framework to assess the impact of habitat versus geographic distance on population differentiation showed that habitat features (at a five km resolution) were equally likely to be correlated with differentiation as geographic distance considered alone. Further differentiation of the geographically disjunct groups is likely to occur in the future. The viability of either group alone is uncertain given their restricted distribution, small population sizes (based on census data and Ne estimates) and isolation. Roost provision and habitat restoration in the geographic region separating the differentiated groups will be fundamental to the recolonization of this area and the reestablishment of connectivity between the regional groups.

6

Empirical assessment of non-invasive population genetics in bats: Comparison of DNA quality from faecal and tissue samples

63%

Boston E. S. M. , Puechmaille S. J. , Scott D. D. , Buckley D. J. , Lundy M. G. , Montgomery I. W. , Prodohl P. A. , Teeling E. C.

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nr 1

EN

Non-invasive population genetics has become a valuable tool in ecology and conservation biology, allowing genetic studies of wild populations without the need to catch, handle or even observe the study subjects directly. We address some of the concerns regarding the limitations of using non-invasive samples by comparing the quality of population genetic information gained through DNA extracted from faecal samples and biopsy samples of two elusive bat species, Myotis mystacinus and Myotis nattereri. We demonstrate that DNA extracted from faeces and tissue samples gives comparable results for frequency based population genetic analyses, despite the occurrence of genotyping errors when using faecal DNA. We conclude that non-invasive genetic sampling for population genetic analysis in bats is viable, and although more labour-intensive and expensive, it is an alternative to tissue sampling, which is particularly pertinent when specimens are rare, endangered or difficult to capture.

7

A taxonomic review of Hipposideros halophyllus, with additional information on H. ater and H. cineraceus (Chiroptera: Hipposideridae) from Thailand and Myanmar

63%

Douangboubpha B. , Bumrungsri S. , Soisook P. , Murray S. W. , Puechmaille S. J. , Satasook C. , Si Si Hla Bu B. , Harrison D. L. , Bates P. J. J.

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2010

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tom 12

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nr 1

EN

Based on recent field surveys in Thailand, Myanmar, and northern peninsular Malaysia, this paper reviews the taxonomy, morphometric and acoustic characters, distribution and ecology of the little known, globally endangered species Hipposideros halophyllus. It lists nine new localities records, including the first from northern and peninsular Thailand, which represent a substantial increase in the species' known range; it confirms the record from northern peninsular Malaysia. In addition, it provides further information on two other small species of the Hipposideros bicolor group, H. ater and H. cineraceus.