Molecular phylogenetic identification of bat reservoirs for Rabies virus

CARBALLO DIEGO; MONTANI MARIA EUGENIA; MARTÍNEZ LEILA; ANTONIAZZI LEANDRO RAUL; CISTERNA DANIEL; BELTRAN FERNANDO; COLOMBO, VALERIA C.

Buenos Aires

Congreso; XXIX International conference of rabies in the americas; 2018

RITA Rabies in the americas

Bats are main reservoirs for Lyssavirus worldwide, constituting an important public health issue since they represent one of the main challenges in rabies control. Different rabies virus (RABV) variants are maintained by non-hematophagous bats in nature which are mainly species-specific. The correct identification of both bat species and RABV variant are essential in epidemiological surveillance to detect new variants as well as spillover events.In this study, we implemented a molecular phylogenetic approach to identify bats at the species level. A total of 15 unknown specimens, belonging to two families, Vespertilionidae (Myotis spp.) and Molossidae (Molossus spp. and Eumops spp.), captured in the province of Santa Fe were studied. Genomic DNA was extracted from patagium or muscle samples. The complete mitochondrial cytochrome-b gene was amplified by PCR and sequenced using both primers. A Bayesian molecular phylogeny was obtained including unknown samples, plus 3 outgroups and 88 additional sequences representing all possible genera and allied species present in the sampling area, retrieved from Genbank.Molecular analysis show that samples belong to three genera: Myotis, Molossus, and Eumops. The three Eumops samples were unambiguously identified as E. bonariensis. Five Myotis samples were identified as M. nigricans, but their possible ascription to M. aff. diminutus is discussed. The 7 remnant samples segregate into two subclades within Molossus molossus, but the possible relationship between one of these clades and M. currentium is examined.This method proved to be useful for the identification of bat species when there is a reliable pre-existing classification. When such classification is absent, this approach combined with morphological data could serve to generate reference sequences to further identify samples remitted in the future.