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Epidemiological, evolutionary and ecological spillover-risk assessment of bat viruses from Atlantic Forest Hotspot

Abstract

Emerging infectious diseases represent a threat to public health worldwide. The emergence and spread of the SARS-CoV-2 in the human population probably originated from ancestral bat viruses evolving into a Pandemic that has caused thousands of deaths worldwide, clearly demonstrating the importance of etiologic agents from bats reservoirs. Before the pandemic, bats were already recognized as important reservoirs of emerging viruses of high mortality and comorbidity in humans, such as the Betacoronaviruses CoV-MERS and CoV-SARS, the Filoviruses such as Ebola and Marburg, and Paramyxoviruses such as Nipah and Hendra. The emergence of zoonotic diseases has been accelerating in the last decades, probably due to anthropogenic actions. The Atlantic Forest (AF) is classified as a "hot spot" for the emergence of infectious diseases due to the high level of biodiversity, habitat of 117 bat species, and the high anthropogenic pressure. Holding the largest urban centers in South America and home to the largest percentage of the Brazilian population (125 mi), the AF also is the most productive land in Brazil, making potential zoonotic hosts from AF essential targets for viral surveillance. Recent data obtained from our studies indicate the presence of different viral groups related to emerging viruses in bats from AF, including Betacoronaviruses, Hantaviruses, Arenaviruses, Influenza viruses, and Paramixoviruses, whose eco-epidemiology, genetic diversity, and zoonotic potential are still poorly known. To deepen the knowledge about the diversity and the possibility of the emergence of distinct viruses from this host and biome, we propose to perform active genomic surveillance in bats from pristine forests, urban and rural areas of the AF, followed by eco-epidemiology, evolution, seroprevalence, and cell tropism evaluation of distinct viruses making possible to analyze the potential for spillover risks of bat-borne viruses. Bat carcasses will be obtained from pristine, urban, and peri-urban forest areas of the São Paulo Metropolitan region and rural areas of the Northwestern part of São Paulo state, an important agricultural and livestock production region. An active surveillance study will also collect blood and oral/rectal swab samples from captured animals. RNA will be extracted from distinct samples and submitted to broad RT-PCR assays targeting specific virus groups such as Coronaviruses, Hantaviruses, Arenaviruses, and Paramyxoviruses. Preliminary phylogenetic data will indicate the samples to be exposed to next-generation sequencing and used in inoculation attempts in cell lines such as Vero, BHK, and Caco. Further tropism tests on other human and animal cells to evaluate the potential of host switch will also be performed. Primary cell culture of specific bat tissues obtained from collaborators or developed in our lab will be used for bat virus isolation attempts. Pseudovirus of Vesicular Stomatitis Virus (VSV) will be produced for virus strains that could not be isolated and used on seroprevalence and tropism assays. Data generated from this study will deepen the eco-epidemiological understanding of potential viral agents emerging from bats in areas with high population density, intense deforestation by human encroachment, and agricultural activity, areas of the most significant theoretical potential for viral transmission events between mammalian species and humans. (AU)

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