key: cord-0788057-8v7851w7
authors: Abreu, Filipe Vieira Santos de; Macedo, Mariana Viana; Silva, Alex Junio Jardim da; Oliveira, Cirilo Henrique de; Ottone, Vinícius de Oliveira; Almeida, Marco Antônio Barreto de; Santos, Edmilson dos; Cardoso, Jader da Cruz; Campos, Aline Scarpellini; Silva, Claudia Maria Dornelles da; Silva, Amanda Gonzales da; Andrade, Miguel de Souza; Bernis, Valéria Magro Octaviano; Bernis Filho, Walter Octaviano; Trindade, Giliane de Souza; Albuquerque, George Rego; Sevá, Anaiá da Paixão; Ribeiro, Bergmann Morais; Teixeira, Danilo Simonini; Campos, Fabrício Souza; Franco, Ana Cláudia; Roehe, Paulo Michel; Oliveira, Danilo Bretas de
title: No evidence of SARS-CoV-2 infection in Neotropical Primates sampled during COVID-19 pandemic in Minas Gerais and Rio Grande do Sul, Brazil
date: 2021-06-18
journal: bioRxiv
DOI: 10.1101/2021.06.17.448890
sha: 7ccad2e04212109acc72d576587a98840795edb2
doc_id: 788057
cord_uid: 8v7851w7

In 2019, a new coronavirus disease (COVID-19) was detected in China. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was capable to infect domestic and captive mammals like cats, tigers and minks. Due to genetic similarities, concern about the infection of Non-Human Primates (NHPs) and the establishment of a sylvatic cycle has grown in the Americas. In this study, neotropical primates (NP) were sampled in different areas from Brazil to investigate whether they were infected by SARS-CoV-2. A total of 89 samples from 51 NP of four species were examined. No positive samples were detected via RT-qPCR, regardless of the NHP species, tissue or habitat tested. This work provides the first report on the lack of evidence of circulation of SARS-CoV-2 in NP. The expand of wild animals sampling is necessary to understand their role in the epidemiology of SARS-CoV-2.

there is a great concern about the risks of SARS-CoV-2 spillback from humans to wildlife, including neotropical

NHPs. This could have at least two impacts: a) establishment of a sylvatic cycle with serious implications on control or eradication efforts, and: b) unforeseeable impact on biodiversity, especially if endangered NHP species become involved. The historical comparison with yellow fever virus (YFV), whose sylvatic cycle was first recognized almost a century ago after a spillback from human to NHPs, exemplifies the potential damage that may be associated to such impacts (Possas et al. 2018 ).

To our knowledge, the only neotropical primate species experimentally infected with SARS-CoV-2 was the marmoset Callithrix jacchus. Infected individuals had pyrexia and viral RNA loads detectable for several days post-infection in oral, nasal and anal swabs, as well as in blood and stool samples (Lu et al. 2020 ).

primates' families or species. Worryingly, in many Brazilian cities where cases of COVID-19 are currently reaching unprecedented levels, Callithrix spp. are adapted to synanthropic environments, often kept as pets and feeding on human foods. Such coexistence might represent a significant potential source of infection (Longa et al. 2011 ). In addition, the rapid dissemination of SARS-CoV-2 currently verified throughout Brazil can lead to the exposure of other NHPs, including wild species.

The present study was carried out in search for serological and virological evidences of SARS-CoV-2 infections in NHPs from urban, sylvatic and rural areas of two Brazilian states, Minas Gerais and Rio Grande do Sul. The current study is part of a wider NHP sampling effort whose main goal is to contribute to the national program for epidemiological surveillance of yellow fever virus.

Neotropical Primates' samplings were performed before (November 2019) and during (July 2020 - Live marmosets were captured with Tomahawk traps, according to the protocol previously described In total, 51 NHPs belonging to two different families (Callitrichidae and Atelidae) and four species (Callithrix penicillata, C. jacchus, Alouatta caraya and A. guariba clamitans) were examined (Table 1) . Seven NHPs were captured before the reported virus introduction (august -november / 2019) and 44 after the introduction (july 2020 to february 2021). Different tissues were analyzed, totalizing 89 samples: 30 of serum, 20 of oral swabs, 19 of liver, 14 of kidney, 3 of lung and 3 of whole blood (Table 1) . No positive samples were detected, regardless of NHP species, tissue, habitat, municipalities or state/biome tested (Table 1) . This is the first study evaluating the natural infection of free-living neotropical NHPs with SARS-CoV-2. We found no evidence of SARS-CoV-2 infection in NHPs from urban, rural or sylvatic habitats regardless of the tissue or species surveyed. Even so, it is important to maintain and expand the surveillance of wildlife, aiming at the early detection of spillover / spillback among human beings and other animals, considering the widespread of SARS-CoV-2 in Brazil.

Since the emergence, reports of natural SARS-CoV-2 infection of several animals have accumulated,

including domestic animals like cats (Sailleau et al. 2020; Carlos et al. 2021 ) and wild captive animals like tigers (McAloose et al. 2020 ) and minks (Hammer et al. 2021) . Minks have also been described as a source of infection for humans (Hammer et al. 2021 ). In addition, experimental infection studies have reported susceptibility of several animals such as ferrets, bat, hamsters Imai et al. 2020) , and mainly Old World NHPs Lu et al. 2020) . As a result, concern about the SARS-CoV-2 spillback for Neotropical Primates -as happened with YFV in the last century and probably with ZIKA in the last decadehas grown (Terzian et al. 2018; Yunes Guimarães et al. 2020 ).

The only report of experimental infection in a neotropical NHP published to date, examined the dynamics of SARS-CoV-2 infection in Callithrix jacchus, the common marmoset. Viral genomes were detected in oral, nasal and anal swabs as well as in serum and feces, despite the lower susceptibility of the species when compared to Old World Primates. (Lu et al. 2020 differences across the twelve binding sites regarded as essential for the ACE2 protein to allow binding of SARS-CoV-2 (Melin et al. 2020; Joyraj Bhattacharjee et al. 2021 ). This may be related to species' differences in susceptibility and, possibly, with our negative results, even in NHPs living in urban areas with high SARS-CoV-2 human incidence. Even so, it is important to investigate whether the new variants of SARS-CoV-2 could increase their infectivity in wild animals.

Brazil is the country with the greatest biodiversity on the planet and concentrates the largest number of NHP species, many of these endangered due to anthropogenic effects such as deforestation which, in addition to drastically reducing habitats, increases the risks of zoonotic transmissions (Paglia et al. 2012; Guthid et al. 2020 ). The explosive spread of SARS-CoV-2 in the country and the close contact between humans and native fauna, provides conditions for spillovers and spillback which increases the need for surveillance for early identification and management of potential harmful effects (Gryseels et al. 2020) . In this sense and despite the small sampling, our study contributes to shedding light on the subject by providing the first results about the absence of circulation of SARS-CoV-2 in Neotropical Primates. It is urgent to expand the sampling of NHP living in different environments, to include other primate species and even other mammals to carry out an active surveillance of wild animals and understand the epidemiology of SARS-CoV-2 in natural environments.

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The authors thanks to all institutions and technicians that contributed and supported the field work. We are especially grateful to Centro de Estudo Multidisciplinar em Conservação Animal IFNMG -Salinas (CEMCA),