Of Animal and Men: The Importance of Animal Environment to Antimicrobial Resistance: A One Health Approach

The contribution of the animal environments to the worsening of the global antimicrobial resistance framework is related to the use of antimicrobials in subtherapeutic doses and, for long periods, establishing ideal conditions for the circulation of resistance genes, which can be transmitted to pathogens adapted to the human microbiota. The study of the animal environment as conducive to the acceleration of resistance evolution is an emerging and critical area for understanding the development and dissemination of resistance genes among the circulating bacteria. The connection between people, animals, and the environment allows us to consider antimicrobial resistance in an approach within the “One Health” concept, which provides a global strategy for expanding collaboration and interdisciplinary communication. This chapter will highlight the emergence of colistin resistance, a great challenge in antimicrobial resistance field. Also, it will focus on some agents included in the priority list of superbugs of the World Health Organization (WHO) or correlated species already identified in veterinary medicine, such as the critical superbugs; priority level 1, Carbapenem-resistant Acinetobacter baumannii, Carbapenem-resistant Pseudomonas aeruginosa, and ESBL-producing Carbapenemic-resistant Enterobacteriaceae; and the high-priority, level 2, methicillin-resistant Staphylococcus aureus (MRSA)

TESTE DE HODGE MODIFICADO EM ÁGAR CLED PARA TRIAGEM DE Proteus mirabilis PRODUTORES DE CARBAPENEMASE

Enterobacteriaceae produtoras de carbapenemase vêm sendo descritas em todo o mundo.  Uma detecção precisa de bactérias produtoras de carpabenemase é necessária pois esta classe de antibióticos é usada no tratamento de infecções severas. A nível laboratorial, o método fenotípico para a detecção de produtores de carbapenemase é o teste de Hodge modificado. Entretanto, algumas enterobactérias tem grande motilidade dificultando a leitura e interpretação dos resultados desta técnica. O objetivo deste estudo foi validar um meio para se obter resultados confiáveis em bactérias com grande motilidade, como é o caso de Proteus mirabilis. O meio ágar Müller-Hinton, preconizado pelo CLSI, foi comparado ao ágar CLED que demostrou ser um bom meio para análise da produção de carbapenemase em Proteus mirabilis suspeitos de produzirem esta enzima embora todos os isolados tenham sido negativos no teste

Isolamento de agentes microbianos a partir de amostras de sangue e umbigo de bezerros mestiços neonatos

Umbilical structures infections can be followed by bacteremic, septicemic infections and death in newborns with passive immunity deficiency. Microorganisms isolated in omphalitis have been also isolated from animals with bacteremia. From september/2002 to september/2003, a research was developed using 44 calves, in order to evaluate bacteremia frequency in newborns and to correlate microorganisms isolated from blood samples and umbilical structures. Blood samples were collected from jugular vein for microorganisms isolation. Twenty four samples were collected in a period of 24 to 48 hours, the other 20 samples from thirty to fifty days after birth. Umbilical structures materials were collected though swabs. Microbial growth occurred in 17 (38,67%) blood samples and in 100% of umbilical samples. Staphylococcus spp., E. coli, Bacillus spp, Pseudomonas spp. and Streptococcus spp were the major isolated microorganisms from blood. All bacteremic animals presented systemic or localized clinical manifestations. The most reported clinical sign was thickness of umbilical structures. Bacillus spp, Enterobacter spp, Micrococcus spp. and E. coli were isolated from umbilical structures. Data confirm a mixed bacterial environment in omphalitis, and suggest a high prevalence of bacteremic infections in newborns calves, pointing out the need of passive immunity transfer through colostrum.Infecção das estruturas umbilicais pode resultar em bacteremia, septicemia e morte em neonatos com falha na imunidade passiva sendo os microrganismos usuais de onfalites isolados freqüentemente em animais com bacteremia. Um estudo foi desenvolvido entre setembro 2002 e setembro 2003 utilizando-se 44 bezerros, visando verificar a freqüência de bacteremia em bezerros neonatos e a correlação entre os agentes isolados a partir de amostras de sangue de estruturas umbilicais. Amostras de sangue foram obtidas por punção da jugular e submetidas à cultura para isolamento de agentes microbianos, sendo 24 obtidas entre 48 e 72 horas e 20 entre o terceiro e o quinto dia após o nascimento. Através de "Swabs" procedeu-se à coleta de material das estruturas umbilicais para a mesma finalidade. Obteve-se crescimento microbiano em 17 (38,67%) amostras de sangue e 100% das amostras de estruturas umbilicais. Os microorganismos mais freqüentes em amostras de sangue foram Staphylococcus sp., E. coli, Bacillus spp, Pseudomonas sp. e Streptococcus. Todos os animais com bacteremia apresentaram manifestações de enfermidade focal ou sistêmica. O sinal clínico mais freqüentemente relacionado com cultura de sangue positiva foi o espessamento das estruturas umbilicais. Bacillus spp, Enterobacter spp, Micrococcus spp. e E. coli foram os microrganismos isolados das estruturas umbilicais. Os dados confirmam uma flora bacteriana mista nos casos de onfalite e sugerem uma prevalência alta de bacteremia em bezerros neonatos, sobretudo aqueles com onfalite, evidenciando a importância de boa transmissão de imunoglobulinas através do colostro

Isolamento de espécies enterobacterianas em Stomoxys calcitrans Isolation of enterobacterial species in Stomoxys calcitrans

Este estudo teve por objetivo relatar o isolamento de espécies enterobacterianas em segmentos de Stomoxys calcitrans coletadas, em propriedades rurais de dois municípios do Estado do Rio de Janeiro, Brasil. Nove das 28 espécies isoladas e identificadas ainda não haviam sido descritas na mosca dos estábulos. Algumas dessas espécies possuem destacado potencial patogênico ao homem e aos animais ou são oportunistas e outras não foram descritas como causadoras de enfermidades de importância médica ou veterinária.<br>This study aimed to report enterobacterial species recovered from body segments of Stomoxys calcitrans collected in dairy farms from counties of Rio de Janeiro State, Brazil. Nine of the isolated and identified species were not yet described in stable flies, according to the worldwide literature. Some of them are known to present pathogenical potential to man and other animals, while others were not described causing diseases of medical or veterinary importance

Characterization of quinolone resistance in Salmonella spp. isolates from food products and human samples in Brazil

Abstract Non-typhoidal salmonellosis is an important zoonotic disease caused by Salmonella enterica. The aim of this study was to investigate the prevalence of plasmid-mediated quinolone resistance in Salmonella spp. and its association with fluoroquinolone susceptibility in Brazil. A total of 129 NTS isolates (samples from human origin, food from animal origin, environmental, and animal) grouped as from animal (n = 62) and human (n = 67) food were evaluated between 2009 and 2013. These isolates were investigated through serotyping, antimicrobial susceptibility testing, and the presence of plasmid-mediated quinolone resistance (PMQR) genes (qnr, aac(6')-Ib) and associated integron genes (integrase, and conserved integron region). Resistance to quinolones and/or fluoroquinolones, from first to third generations, was observed. Fifteen isolates were positive for the presence of qnr genes (8 qnrS, 6 qnrB, and 1 qnrD) and twenty three of aac(6')-Ib. The conserved integron region was detected in 67 isolates as variable regions, from ±600 to >1000 pb. The spread of NTS involving PMQR carriers is of serious concern and should be carefully monitored

Occurrence of Shiga-toxigenic Escherichia coli in Stomoxys calcitrans (Diptera: Muscidae)

This study aimed to verify the occurrence of Shiga toxin-producingEscherichia coli (STEC) strains in three distinct anatomic parts of the stable fly Stomoxys calcitrans by multiplex polymerase chain reaction (PCR Multiplex). According to the results obtained,E. coli was identified in 19.5% of the stable flies. Shiga toxin genes were detected in 13% of the E. coli isolated, most frequently from the surface, followed by abdominal digestive tract and mouth apparatus of insects, respectively. This is the first study to detect presence of STEC in Stomoxys calcitrans in Brazil; it has also revealed the potential role of stable flies as carriers of pathogenic bacterial agents

Bacterial diversity and detection of resistance genes to broad-spectrum betalactams in dairy family farm soils

Bovine mastitis is a complex disease that brings great losses to the dairy producer. The microbial diversity of the soils, as well as the presence of resistance genes in the environment directly influence the maintenance of mastitis in the farm. The objective of this work was to analyze the bacterial diversity in pasture soils of a dairy family farm, detecting enterobacteria that may be involved in the etiology of bovine mastitis, and to detect genes that encode broad-spectrum betalactamases in these soils. Twelve soil samples, representative of different areas of the farm located in the municipality of Barra do Piraí, Rio de Janeiro, were collected at different times of the year. Total DNA was extracted from the samples, gene amplified by Nested-PCR and then the amplification products were separated by DGGE (Denaturing Gradient Gel Electrophoresis). With the DGGE it was possible to construct dendograms that effectively represented the bacterial diversity of these soils. Eight of the soil samples were used to amplify the genes encoding the betalactamase enzymes TEM (blaTEM gene), SHV (blaSHV gene) and CTX (blaCTXM gene). In three of the eight soil samples, the blaSHV gene was found to be present. The blaTEM and blaCTX-M genes were not detected in any of the samples. The detection of genes encoding broad-spectrum betalactamases in dairy cattle pasture soils is of concern, because the transfer of gene material between pathogenic and non-pathogenic bacteria in this environment is a reality

Mapeamento do perfil de resistência e detecção do gene mecA em Staphylococcus aureus e Staphylococcus intermedius oxacilina-resistentes isolados de espécies humanas e animais Resistance pattern and detection of mecA gene in oxacillin-resistant isolates of Staphylococcus aureus and Staphylococcus intermedius from animal and human samples

Espécies de Staphylococcus spp resistentes a antimicrobianos representam um problema cosmopolita, sendo o controle de sua disseminação um importante desafio. O perfil de resistência a antimicrobianos de isolados de Staphylococcus aureus e Staphylococcus intermedius em amostras clínicas humanas e animais foi avaliado através do método de difusão em disco, no qual foi possível detectar um elevado nível de resistência à ampicilina e à penicilina. A avaliação da resistência à oxacilina, devido à heterogeneidade de resposta do gênero estudado, foi desenvolvida também através das seguintes técnicas: difusão em ágar modificado, ágar screen e microdiluição em caldo, e posteriormente correlacionada com a detecção do gene mecA, pela técnica de PCR, nas amostras consideradas resistentes em pelo menos um dos testes utilizados. A correlação entre os resultados obtidos nos testes fenotípicos com a presença do gene de resistência, considerado um método de referência, foi utilizada para validar a sensibilidade destes. De um total de 80 amostras avaliadas, 28 apresentaram resistência à oxacilina, sendo possível detectar a presença do gene de resistência mecA em 12 destas amostras. Os testes de suscetibilidade à oxacilina apresentaram sensibilidade superior a 50,0%, sendo a difusão em disco simples e o ágar screen considerados mais sensíveis, e a difusão em disco modificada, o de menor sensibilidade.Antimicrobial resistant Staphylococcus species represent an important cosmopolitan problem, and its spreading control is a significative challenge. Resistance pattern of Staphylococcus aureus and Staphylococcus intermedius species isolated from animals and humans clinical samples to different antibiotics was evaluated through disk diffusion method, where ampicillin and penicillin presented the highest level of resistance. The evaluation of the resistance to oxacillin, due to the heterogeneity of the response of the studied genus was carried out through the following tests: modified agar diffusion, agar screen and microdilution, and further correlation with the detection of mecA gene in samples that showed resistance in at least one of the susceptibility tests used. The correlation between the results obtained from phenotypic methods and the detection of resistance gene, considered as a reference method, was used in order to validate its sensitivity. Eighty clinical staphylococcal isolates (29 human and 51 animal isolates) were evaluated, 28 were oxacillin-resistant, mecA gene being detected in 12 samples. Susceptibility assessment tests to oxacillin presented above 50% of specificity, disk diffusion and agar screen being the most sensitive one, while modified disk diffusion presented the lowest sensibility rate. Ampicillin and penicillin presented the highest level of resistance