Virus detection in snakes held captive in Butantan Institute Plantels

• Grant number: 19/12303-4
• Support Opportunities: Regular Research Grants
• Duration: August 01, 2020 - January 31, 2023
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Viviane Fongaro Botosso
• Grantee: Viviane Fongaro Botosso
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Associated researchers: Carla Lilian de Agostini Utescher ; Danielle Bruna Leal de Oliveira ; Edison Luiz Durigon ; Kathleen Fernandes Grego ; Sávio Stefanini Sant Anna ; Selma Maria de Almeida Santos

Abstract

The (WHO) included snakebites accidents in the list of neglected tropical diseases mainly affecting rural and low-income populations, with limited access to education and health systems. WHO estimates that approximately 5 million of snake bites occur annually, resulting in up to 2.7 million of poisonings, with 81,000 to 138,000 deaths and 400,000 amputations or other type of permanent physical damage per year, thus presenting high morbidity and mortality rates. In Brazil between 2000 and 2017 more than 470,000 cases of snakebites accidents were reported, and the disease has been included on the Compulsory Notification list (LNC) of Brazil since 2010. The most common accidents in our country are caused by snakes of the genus: Bothrops; Crotalus; Lachesis and Micrurus. The most efficient treatment of this important disease is antivenom heterologous Serum. Instituto Butantan (IBUT) is the main producer of these immunobiologicals and is the only national producer against Lachesis and Micrurus accidents. Aiming the production of all these sera the IBut maintains and reproduces snakes for poison extraction, considered the initial raw material for the production of the sera. The poisons produced are also used in several research lines, including, for example, toxinology, pathophysiology, immunology and studies of bio-prospecting. In addition to this activity, the Institute maintains other snakes used in different areas of research and cultural diffusion, including the preservation program of Jararaca-Ilhoa, a brazilian snake, threatened by extinction. Several prophylactic measures are taken before the snakes are incorporated into the bioteriumm to prevent the entry and dissemination of pathogens and, consequently, to improve the health and welfare of the animals, and enhance the quality of the obtained poisons. However, all these measures are still unable to detect or even prevent virus entry, because it is often necessary several months before clinical changes caused by these organisms can be noticed. Thus, the present project aims to implement the methodology of detection and identification of viruses in snakes maintained in captivity in the laboratories of Instituto Butantan, aiming the optimization and improvement of the protocols of management and conservation of the animals. We emphasize that this area of research has grown a lot in the last decade, including the description of new species of viruses and that in Brazil these studies are very rare.Blood samples, cloacal and tracheal swabs, as well as other clinical samples collected during pertinent surgical procedures, will be collected from different species while in quarantine period, before being incorporated into the bioterium, as well as from animals of the bioterium presenting clinical symptomatology suggestive of viral infectious disease (as respiratory changes and the presence of visceral or subcutaneous masses). In addition animals dying without confirmed clinical diagnosis will have fragments of all tissues sampled, such as lung, venom gland, liver and brain, for histopathological analysis and for viruses researcher. These fragments will be subjected to extraction of nucleic material, followed by obtainment of cDNA (when necessary). Virus detection will be carried out by the PCR technique for the following families: Astroviridae, Arenaviridae, Coronaviridae, Flaviviridae, Filoviridae, Paramyxoviridae, Caliciviridae, Adenoviridae, Herpesviridae and genera: Alphavirus, Lyssaviru, Hantavirus, Enterovirus, Phlebovirus, Nipahvirus, influenza, Orbivrus. The amplicons will be subjected to the sequencing reaction by the Sanger method to confirm and characterize the detected viruses. We intend, at the end of this Project, to implement the methodology routinely at the bioteriums, improving the management and care of the animals. (AU)