Abstract
With more than 1400 chiropteran species identified to date, bats comprise one-fifth of all mammalian species worldwide. Many studies have associated viral zoonoses with 45 different species of bats in the EU, which cluster within 5 families of bats. For example, the Serotine bats are infected by European Bat 1 Lyssavirus throughout Europe while Myotis bats are shown infected by coronavirus, herpesvirus and paramyxovirus. Correct host species identification is important to increase our knowledge of the ecology and evolutionary pattern of bat viruses in the EU. Bat species identification is commonly determined using morphological keys. Morphological determination of bat species from bat carcasses can be limited in some cases, due to the state of decomposition or nearly indistinguishable morphological features in juvenile bats and can lead to misidentifications. The overall objective of our study was to identify insectivorous bat species using molecular biology tools with the amplification of the partial cytochrome b gene of mitochondrial DNA. Two types of samples were tested in this study, bat wing punches and bat faeces. A total of 163 bat wing punches representing 22 species, and 31 faecal pellets representing 7 species were included in the study. From the 163 bat wing punches tested, a total of 159 were genetically identified from amplification of the partial cyt b gene. All 31 faecal pellets were genetically identified based on the cyt b gene. A comparison between morphological and genetic determination showed 21 misidentifications from the 163 wing punches, representing ~12.5% of misidentifications of morphological determination compared with the genetic method, across 11 species. In addition, genetic determination allowed the identification of 24 out of 25 morphologically non-determined bat samples. Our findings demonstrate the importance of a genetic approach as an efficient and reliable method to identify bat species precisely.
Highlights
All bat species and their roosts are legally protected in France and in Europe by national and international legislation due to the significant decrease in their populations over the last few decades
The aim of this study was 1) to optimize the rapid Polymerase Chain Reaction (PCR) method previously described in Lopez-Oceja et al (2016) with the new universal cyt b primers to identify autochthonous bat species from different types of bat sample, namely guano and wing punches tested for the first time; 2) to genetically determine bats in France and 3) to compare the morphological and genetic species identification of bat carcasses submitted for rabies diagnosis in 2018 and 2019
Of the 18 bat carcasses morphologically not determined, we reported the identification of 6 species belonging in the Vespertilionidae family with P. pipistrellus (n = 6), P. kuhlii (n = 7), M. daubentonii (n = 1), R. hipposideros (n = 1), Hypsugo savii (n = 2) and P. pipistrellus or P. pygmaeus (n = 1)
Summary
All bat species and their roosts are legally protected in France and in Europe by national and international legislation due to the significant decrease in their populations over the last few decades. 51 bat species have been identified in Europe of which 35 occur in France, including one new cryptic species (Myotis crypticus) recently identified in France [3, 4]. Insectivorous bats in Europe are divided into four different families: Rhinolophidae, Vespertilionidae, Molossidae and Miniopteridae [7, 8]. Some studies have suggested a possible association between some bat species and micro-organisms, including more than 200 viruses, bacteria, parasites and pathogenic fungi. Common in France, has been reported with the presence of alphacoronaviruses [13], herpesviruses and with Yersinia bacteria [14]
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