Genetic identification of bat species for pathogen surveillance across France

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.

Fig 1. Phylogenetic tree of the partial cytochrome b (cyt b) gene of 52 referenced sequences and 25 bat sequences representing 15 autochthonous bat species.

B.barb: Barbastella barbastellus, E.nils: Eptesicus nilssonii, E.ser: Eptesicus serotinus, H.s: Homo sapiens, H.savi: Hypsugo savii, M.al: Myotis alcathoe, M.bech: Myotis bechsteinii, M.bly: Myotis blythii, M.br: Myotis brandtii, M.c: Myotis capaccinii, M.daub: Myotis daubentonii, M.das: Myotis dasycneme, M.em: Myotis emarginatus, M.esc: Myotis escalerai, M.myo: Myotis Myotis, M.mys: Myotis mystacinus, M. nat: Myotis nattereri, M.schr: Myotis schreibersii, N.leis: Nyctalus leisleri, N.noct: Nyctalus noctula, P.aur: Plecotus auritus, P.aust: Plecotus austriacus, P.kuh: Pipistrellus kuhlii, P.pip: Pipistrellus pipistrellus, P.pyg: Pipistrellus pygmaeus, P.nath: Pipistrellus nathusii, R.fer: Rhinolophus ferrumequinum, R.hip: Rhinolophus hipposideros, R.meh: Rhinolophus mehelyi, S.a: Sorex araneus, V.mur: Vespertilio murinus.