Analysis of the antimicrobial resistance profile in bacteria isolated from necropsied animals at the Federal Rural University of Rio de Janeiro

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2022-05-10T18:38:35Z No. of bitstreams: 1 2022 - Leticia Baptista Pinto.pdf: 1572478 bytes, checksum: 9b8b1148e65527a8c43819d714e79dd8 (MD5) Made available in DSpace on 2022-05-10T18:38:35Z (GMT). No. of bitstreams: 1 2022 - Leticia Baptista Pinto.pdf: 1572478 bytes, checksum: 9b8b1148e65527a8c43819d714e79dd8 (MD5) Previous issue date: 2022-03-18 CAPES - Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior FAPERJ - Funda??o Carlos Chagas Filho de Amparo ? Pesquisa do Estado do Rio de Janeiro The emergence and spread of antimicrobial resistance is one of the three main threats to Public Health in the 21st century and must be analyzed in an integrated One Health approach, as it is a health risk shared by people, animals and the environment. Despite understanding the multifactorial origin of antimicrobial resistance, little is known about the contribution of environments aimed at the production, maintenance and care of animals in disseminating this phenomenon. Among these, the necropsy space represents a point of cohesion, being a place of extreme relevance for research and understanding of the circulation of bacterial microbiota and its resistance genes. The present study evaluated the occurrence of superbugs in samples of animals necropsied at the Federal Rural University of Rio de Janeiro, considering the priority criteria established by the World Health Organization (WHO). Of the 198 samples collected from 45 animals, being 20 companion animals, 20 production animals, and three wild ones, 325 strains were isolated, of which 51,38% (167/325) were Enterobacterales, 31,69% (103/325) Staphylococcus spp., 12,62% (41/325) Enterococcus spp., 2,46% (8/325) Streptococcus spp. and 1,85% (6/325) BGNNF. MALDI-TOF proved to be an efficient tool for bacterial identification, especially in Enterococcus spp. and Enterobacterales. The agreement between biochemical, proteomic and genotypic techniques in identifying Staphylococcus spp. was 80,58%, which confirms the importance of the association between different diagnostic methods for the characterization of this genus, leading to the correct direction of the resistance analysis. 8,74% (9/103) of Staphylococcus spp. showed phenotypic resistance indicative of PBP2a production, with detection of the mecA gene in all strains. Phenotypic resistance to vancomycin was evidenced in E. faecalis, with detection of the vanB gene. In 29,13% (30/103) of Staphylococcus spp. there was detection of the blaZ gene. 11,98% (20/167) of enterobacteria showed resistance to beta-lactams, mediated by the ESBL production, in the screening antibiogram and 80% (16/20) of them were positive in the confirmatory test. The search for genes encoding ESBL revealed the presence of blaSHV in 10,18% (17/167), blaTEM in 6,59% (11/167) and blaCTX-M-1 in 4,19% (7/167). There was no detection of carbapenemase-producing strains. No mcr genes were detected. These results reveal species characterized as critical superbugs in the necropsy environment and reinforce the need to monitor these strains in the veterinary environment, not only for the adoption of adequate control and treatment measures for the animals but also for the implementation of safe protocols for the disposal of their carcasses. O surgimento e a dissemina??o da resist?ncia aos antimicrobianos ? uma das tr?s principais amea?as ? Sa?de P?blica no s?culo XXI, e deve ser analisado em uma abordagem integrada de Sa?de ?nica, por se tratar de um risco ? sa?de compartilhado por pessoas, animais e meio ambiente. Apesar da compreens?o a respeito da origem multifatorial da resist?ncia antimicrobiana, pouco se sabe sobre a contribui??o dos ambientes voltados a produ??o, manuten??o e cuidados de animais na dissemina??o desse fen?meno. Dentre estes, o espa?o de necropsia representa um ponto de coes?o, sendo um local de extrema relev?ncia para pesquisa e compreens?o da circula??o da microbiota bacteriana e seus genes de resist?ncia. O presente estudo avaliou a ocorr?ncia de superbact?rias em amostras de animais necropsiados na Universidade Federal Rural do Rio de Janeiro, considerando os crit?rios de prioridade estabelecidos pela Organiza??o Mundial de Sa?de (OMS). Das 198 amostras coletadas de 45 animais, sendo 20 animais de companhia, 20 de produ??o e 3 selvagens, foram isoladas 325 cepas, das quais 51,38% (167/325) foram Enterobacterales, 31,69% (103/325) Staphylococcus spp., 12,62% (41/325) Enterococcus spp., 2,46% (8/325) Streptococcus spp. e 1,85% (6/325) BGNNF. O MALDI-TOF mostrou-se uma ferramenta eficiente para identifica??o bacteriana, principalmente em Enterococcus spp. e Enterobacterales. A concord?ncia entre as t?cnicas bioqu?mica, prote?mica e genot?pica na identifica??o de Staphylococcus spp. foi de 80,58%, o que confirma a import?ncia da associa??o entre diferentes m?todos diagn?sticos para a caracteriza??o nesse g?nero, levando ao direcionamento correto da an?lise de resist?ncia. 8,74% (9/103) dos Staphylococcus spp. apresentaram resist?ncia fenot?pica indicativa de produ??o de PBP2a, com detec??o do gene mecA em todas as cepas. Em 29,13% (30/103) dos Staphylococcus spp. houve detec??o do gene blaZ. Foi evidenciada resist?ncia fenot?pica ? vancomicina em E. faecalis, com detec??o do gene vanB. 11,98% (20/167) das enterobact?rias apresentaram resist?ncia aos beta-lact?micos, mediada pela produ??o de ESBL, no antibiograma de triagem e 80% (16/20) delas foi positiva no teste confirmat?rio. A pesquisa dos genes que codificam ESBL revelou a presen?a de blaSHV em 10,18% (17/167), blaTEM em 6,59% (11/167) e blaCTX-M-1 em 4,19% (7/167). N?o houve detec??o de cepas produtoras de carbapenemases. N?o foram detectados genes mcr. Esses resultados revelam a ocorr?ncia de esp?cies caracterizadas como superbact?rias cr?ticas pela OMS em ambiente de necropsia e refor?am a necessidade de monitoramento dessas cepas no ambiente veterin?rio n?o apenas para a ado??o de medidas de controle e tratamento adequados dos animais, mas tamb?m para a implementa??o de protocolos seguros para o descarte de suas carca?as.