Phylogenetic inferences reveal multiple intra- and interhost genetic diversity among bat rabies viruses circulating in northeastern Brazil

Background: Rabies, a lethal viral zoonotic disease, remains a significant global public health concern. In northeastern Brazil, in particular, its epidemiology is complex and dynamic, characterized by the presence of several reservoirs associated with human rabies infection.

Methods: This study, conducted from June 2022 to July 2023, was part of a passive epidemiological surveillance initiative under Brazil's National Rabies Surveillance Program. It investigated the presence of Rhabdovirus (RhabV) in 356 postmortem chiropteran brain samples using three diagnostic techniques for rabies and conducted an evolutionary study on both pan-RhabV- and pan-LYSSAV-positive PCR samples. The samples were collected from 20 bat species and different locations in the State of Ceará, an endemic region for the rabies virus (RABV). Rabies-positive samples were further explored through Bayesian, genetic distance mapping and recombination analyses.

Results: From a total of 356 samples collected, 43 (12.07%) were positive for direct immunofluorescence (DIF) and 40 (11.23%) for mouse intracerebral inoculation (MIT) tests. Among the positive results, 40 samples were confirmed by both DIF and MIT, while 13 (3.65%) had inconclusive results for one or both techniques. Molecular assays identified 38 rabies-positive samples (10.67%). Members of the Molossidae and Phyllostomidae families had the highest prevalence, highlighting the role of insectivorous and frugivorous bats in the cycle and dynamics of rabies transmission. Phylogenetic reconstructions revealed three distinct and well-supported clusters and clades, indicating the cocirculation of different RABV lineages in the region and shedding light on both intra- and interhost diversity. We also demonstrated genetic distance among the RABV clusters and inferred that their common ancestor originated in Europe, later diversifying across continents. No recombination breakpoints were identified.

Conclusions: This study highlights the dynamic nature of RABV evolution within individual bat hosts, contributing to the understanding of the genetic diversity of RABV variants found in several bat species in northeastern Brazil. This study provides crucial insights into viral transmission dynamics within and between different host species and is essential for designing effective rabies control and prevention strategies tailored to endemic regions.