Para avaliar um sistema integrado de aquicultura foram realizadas análises microbiológicas da água utilizada neste sistema e determinada a incidência e resistência antimicrobiana dos enteropatógenos no ecossistema relacionado. As amostras de água testadas apresentaram 32,9% de taxas de coliformes fecais (≤1.600/100mL), de acordo com a OMS para piscicultura em águas residuais. Salmonella spp. foram detectadas em 14,5% das amostras. De um total de 33 cepas, 15,1% eram resistentes a um ou dois antimicrobianos testados e resistência a múltiplas drogas não foi observada. Aeromonas spp. foram identificadas em 91,6% das amostras. De um total de 416 cepas, resistência a uma classe de antimicrobianos foi observada em 66,3% e a multirresistência às drogas em 37,7%. Na avaliação da virulência dos isolados de Aeromonas hydrophila, 85,3% das cepas apresentaram Beta-hemólise nos três diferentes tipos de eritrócitos empregados e 99,1% nos eritrócitos de coelho e cavalo, sendo possível a caracterização através da PCR do gene aerA e lip, em 100% das amostras. Os resultados obtidos apontam para a relevância quanto às vantagens da implementação de um sistema integrado, disponibilizando alimentos com custo reduzido, porém este sistema necessita de um controle rígido e efetivo para que estes produtos não constituam veículos para a disseminação de doenças. / To evaluate an integrated aquaculture system, microbiological analyses of water used in this system were carried out and the incidence and antimicrobial resistance of enteropathogens were determined in the related ecosystem. Water samples tested had 32.9% of fecal coliforms rates (≤1600/100mL) in accordance with WHO for psiculture in wastewater. Salmonella spp. were detected in 14.5% of the samples. From a total of 33 strains, 15.1% were resistant to one or two antimicrobial drugs tested and multidrug-resistance was not observed. Aeromonas spp. were identified in 91.6% of the samples. From a total of 416 strains, resistance to one antimicrobial class was observed in 66.3% and multidrug-resistance in 37.7%. In relation to virulence factors of Aeromonas hydrophila, 85.3% of the strains showed beta-hemolysis in three different types of erythrocytes and 99.1% in horse and rabbit erythrocytes. It was possible to characterize by PCR assay, the genes aerA and lip in 100% of the strains. The results indicate the relevance of the benefits of implementing an integrated system, providing food with reduced cost, but this system requires a strict and effective control so that these products do not constitute a vehicle for the spread of disease.

Mycoplasma hyopneumoniae est l’agent causal de la pneumonie enzootique. On le retrouve dans plusieurs élevages de porcs à travers le monde. Même si ce micro-organisme est présent dans plusieurs troupeaux canadiens, peu d’informations sont présentement disponibles sur les isolats québécois. Un total de 160 poumons de porcs possédant des lésions de pneumonie ont été récupérés à l’abattoir, mis en culture et testés par PCR pour M. hyopneumoniae et Mycoplasma hyorhinis. D’autres pathogènes bactériens communs du porc et les virus du syndrome reproducteur et respiratoire porcin (VSRRP), de l’influenza et le circovirus porcin de type 2 (CVP2) ont été également testés. Quatre-vingt-dix pourcent des échantillons étaient positifs pour M. hyopneumoniae et 5.6% l’étaient seulement pour M. hyorhinis. Dans ces échantillons positifs pour M. hyopneumoniae, la concentration de ce mycoplasme variait de 1.17 x 105 à 3.37 x 109 génomes/mL. Vingt-cinq poumons positifs en culture ou par PCR en temps réel pour M. hyopneumoniae ont été sélectionnés, parmi ceux-ci 10 étaient en coinfection avec Pasteurella multocida, 12 avec Streptococcus suis, 9 avec CVP2 et 2 avec le VSRRP. Les analyses des nombres variables de répétitions en tandem à de multiples loci (MLVA) et PCR-polymorphisme de longueur de fragments de restriction (PCR-RFLP) de M. hyopneumoniae ont démontré une forte diversité des isolats de terrain. Par contre, il semble y avoir plus d’homogénéité à l’intérieur d’un même élevage. L’analyse MLVA a également démontré que près de la moitié des isolats possédaient moins de 55% d’homologie avec les souches vaccinales et de référence utilisées dans la présente étude. L’absence d’amplification du locus 1 de M. hyopneumoniae en MLVA a été significativement associée à une baisse de la concentration de bactérie et de la sévérité des lésions. Pour tous les isolats de M. hyopneumoniae, des concentrations minimales inhibitrices (CMI) de faibles à intermédiaires ont été obtenues envers tous les antimicrobiens testés. Les isolats possédant des CMI intermédiaires envers les tétracyclines, les macrolides et les lincosamides ont été testés pour la présence des gènes de résistance tetM, ermB et pour des mutations ponctuelles dans les gènes des protéines L4, L22 et de l’ARNr 23S. Aucun de ces gènes n’a été détecté mais la mutation ponctuelle G2057A a été identifiée. Cette mutation est responsable de la résistance intrinsèque de M. hyopneumoniae face aux macrolides à 14 carbones. Ces résultats indiquent qu’il ne semble pas y avoir de résistance acquise aux antimicrobiens parmi ces isolats. En conclusion, cette recherche a permis d’obtenir de nouvelles données scientifiques sur les isolats québécois de M. hyopneumoniae. / Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, is present in swine herds worldwide. However, little is known about the prevalence of this microorganism in Quebec and Canadian herds. A total of 160 swine lungs with lesions suggestive of enzootic pneumonia were recovered from two slaughterhouses. They were cultured and tested by PCR for M. hyopneumoniae and Mycoplasma hyorhinis and for the porcine reproductive and respiratory syndrome virus (PRRSV), the influenza virus, and the porcine circovirus type 2 (PCV2). Samples were also cultured for other commonly encountered swine pathogenic bacteria. Ninety percent of the samples were positive for M. hyopneumoniae (Real-time PCR) whereas 5.6% were positive only for M. hyorhinis (PCR). The concentration of M. hyopneumoniae in these positive samples varied between 1.17 x 105 and 3.37 x 109 genomes/mL Among 25 selected M. hyopneumoniae positive lungs (culture or real-time PCR), 10 demonstrated a co-infection with Pasteurella multocida, 12 with Streptococcus suis, 9 with PCV2 and 2 with the PRRS virus. Multiple loci variable number of tandem repeats analysis (MLVA) and PCR-restriction fragments length polymorphism (PCR-RFLP) analyses showed a high diversity among field M. hyopneumoniae isolates. However, there seemed to be greater homogeneity within the same herd. The MLVA analysis also demonstrated that almost half of the field isolates presented less than 55% homology with the vaccine and reference strains used in our study. The absence of amplification of one locus (locus 1) of M. hyopneumoniae was significantly associated with a lower number of bacteria and a lower severity of lung lesions. All M. hyopneumoniae isolates showed low to intermediate MICs against the antimicrobials tested. Cultures with intermediate MICs for tetracyclines, macrolides and lincosamides were tested for the presence of previously described resistance genes tetM, ermB and L4, L22 and 23S rRNA point mutations. None of these genes were found, although one point mutation, G2057A, was identified. This mutation is responsible for the intrinsic resistance of M. hyopneumoniae against 14-membered macrolides. These results indicate that there is probably no acquired antimicrobial resistance within these isolates. This research provides new scientific data on M. hyopneumoniae isolates from Canada.

Introdução. A resistência bacteriana a antibióticos é aceita como um dos maiores problemas de saúde pública. As tetraciclinas são antibióticos de amplo espectro, e após seu uso indiscriminado observou-se o surgimento de bactérias resistentes, levando médicos e veterinários a diminuírem seu uso. Objetivos. Verificar o perfil de sensibilidade a tetraciclinas em isolados clínicos e ambientais de Escherichia coli, Klebsiella pneumoniae e Aeromonas spp., bem como pesquisar os principais genes tet associados à resistência a esta classe de antibióticos e determinar a potencial forma de disseminação destes genes através da caracterização de seu ambiente genético. Material e Métodos. Os perfis de sensibilidade à tetraciclina (TET), doxiciclina (DOX), minociclina (MIN) e tigeciclina (TGC) de 572 isolados foram obtidos através das técnicas de Disco-Difusão e Concentração Inibitória Mínima. Os isolados não-sensíveis à tetraciclina foram submetidos a reações de PCR para pesquisa de grupos Inc, genes tet e para a caracterização de seu ambiente genético pela pesquisa das integrases de classes 1, 2, 3 e 4, e dos elementos genéticos móveis Tn1721, IS26, Tn10 e ISAS5. Perfis de similaridade genética dos isolados foram obtidos através das técnicas de ERIC-PCR e PFGE. Após análise destes resultados 33 cepas foram selecionadas para as técnicas de S1-PFGE e transformação. Resultados. A partir dos 572 isolados 18,5 por cento foram resistentes à TET, 13,5 por cento à DOX, 8 por cento à MIN e nenhum à TGC. Vinte e dois por cento dos isolados clínicos e 16,3 por cento ambientais foram resistentes à TET. Os genes codificadores de bomba de efluxo tet(A), tet(B), tet(C), tet(D) e tet(E), foram observados em 25,5 por cento , 33 por cento , 6,5 por cento , 18,9 por cento e 23,5 por cento dos isolados, respectivamente. Noventa e cinco por cento, 100 por cento , 100 por cento e 4,5 por cento das cepas carreando o gene tet(A), tet(B), tet(D) e tet(E), foram não-sensíveis à DOX, nesta ordem. Resistência à MIN foi observada em 4,2 por cento , 78,8 por cento e 100 por cento dos isolados carreando tet(A), tet(B) e tet(D), respectivamente. O gene tet(A) estava associado a Tn1721, tet(B) à Tn10 e tet(C) e tet(D) à IS26. Nenhuma das integrases pesquisadas estavam associadas aos genes tet detectados. Os grupos IncF, IncFIB e IncA/C foram observados em 54,8 por cento , 41,1 por cento e 28,7 por cento dos isolados, respectivamente. Uma cepa de Aeromonas spp. carreava um plasmídio do grupo IncP. Através dos perfis de similaridade genética foi observado que dentre os isolados hospitalares de K. pneumoniae houve a ocorrência de perfis genéticos idênticos, no entanto nos demais isolados do estudo os perfis genéticos observados eram distintos. Das 33 cepas selecionadas para os experimentos de linearização plasmidial e de transformação, 8 foram transformadas com sucesso, nas quais foi observada a presença dos genes tet em plasmídios. Conclusões. Uma baixa porcentagem de resistência à TET foi detectada. Verificou-se que a TGC foi a tetraciclina mais ativa, seguida da MIN. Os genes tet(A) e tet(B) foram os mais prevalentes. Todas as cepas carreando tet(B) e tet(D) foram não-sensíveis a DOX e MIN. Plasmídios dos grupos IncF, FIB e A/C foram os mais detectados neste estudo. Os resultados sugerem que os genes tet(A), (B), (C) e (D) são disseminados por meio de plasmídios e estão associados aos transposons Tn1721, IS10 e IS26. Estudos adicionais com isolados mais recentes e outros gêneros bacterianos são necessários, para contribuir com informações da resistência bacteriana a tetraciclinas. / Introduction. The antibiotic resistance is accepted as one of the major problems for public health. Tetracyclines are broad spectrum antibiotics, and its indiscriminate use promoted the emergence of resistant bacteria, leading physicians and veterinarians to decrease its use. Objectives. Verify the susceptibility of clinical and environmental isolates of Escherichia coli, Klebsiella pneumoniae and Aeromonas spp. to tetracyclines, and also search for the main tet genes associated with resistance to these antibiotics and determine the potential mechanism of tet genes dissemination by characterizing their genetic context. Material and Methods. Disk-Diffusion and Minimum Inhibitory Concentration tests were carried out in 572 isolates using tetracycline (TET), doxycycline (DOX), minocycline (MIN) and tigecycline (TGC). PCR was carried out in TET non-susceptible isolates for the detection of Inc groups, tet genes and its genetic context determination through the search of classes 1, 2, 3, and 4 integrases, and Tn1721, Tn10, IS26 and ISAS5 mobile genetic elements. Genetic similarities patterns were determined by ERIC-PCR and PFGE techniques. After analyzing the results 33 strains were selected for the S1-PFGE and transformation experiments. Results. From 572 isolates, 18.5 per cent were TET-resistant, 13.5 per cent DOX-resistant, 8 per cent MIN-resistant and none resistant to TGC. Twenty-two per cent and 16.3 per cent of clinical and environmental isolates were TET-resistant, in that order. Genes tet(A), tet(B), tet(C), tet(D) and tet(E), coding for efflux pump mechanism, were found in 25.5 per cent , 33 per cent , 6.5 per cent , 18.9 per cent and 23.5 per cent of the isolates, respectively. Ninety-five per cent, 100 per cent , 100 per cent and 4.5 per cent of the isolates carrying tet(A), tet(B), tet(D) and tet(E) were non-susceptible to DOX, respectively. Resistance to MIN was observed in 4.2 per cent , 78.8 per cent and 100 per cent of isolates carrying tet(A), tet(B) and tet(D), in that order. The gene tet(A) was associated with Tn1721, tet(B) with Tn10, and tet(C) and (D) with IS26. None of the searched integrases were associated with the tet genes detected. Groups IncF, IncFIB and IncA/C were respectively observed in 54.8 per cent , 41.1 per cent and 28.7 per cent of the isolates. One Aeromonas spp. was carrying an IncP plasmid. The genetic similarities patterns demonstrated that there were identical genetic patterns among the hospital K. pneumoniae isolates, however all the remaining isolates possessed distinct genetic patterns. Of the 33 strains selected for plasmid linearization and transformation experiments, 8 were successfully transformed, in which the presence of tet genes in plasmids were observed. Conclusions. A low level of tetracycline resistance was detected. TGC was the most active tested antibiotic, followed by MIN. Genes tet(A) and tet(B) were the most prevalent among the isolates. All strains carrying tet(B) and tet(D) were non-susceptible to DOX and MIN. Groups IncF, IncFIB and IncA/C were the most detected in this study. The results suggest that tet(A), (B), (C) and (D) are disseminated by plasmids and are associated with Tn1721, Tn10 and IS26. Additional studies assembling recent isolates and other genera are necessary in order to contribute with information about the bacteria resistance to tetracyclines.

Introdução. A resistência bacteriana é facilitada pela pressão seletiva do uso de antimicrobianos na clínica e em outras atividades, como a agricultura e pecuária, além de poder ser disseminada para a natureza por meio do lançamento inadequado do esgoto ou pela aplicação do lodo de esgoto na agricultura. As -lactamases de espectro estendido (ESBL) são uma das formas mais prevalentes de resistência em Gram negativos no mundo, e seus genes codificadores são disseminados por meio de diversos elementos genéticos, principalmente transposons e integrons mobilizados para plasmídios. Objetivo. Identificar e caracterizar genes codificadores de ESBL, bem como suas prováveis formas de mobilização, em enterobactérias isoladas de fontes ambientais e clínicas. Material e Métodos. Quarenta e cinco cepas isoladas de um hospital público em 2004 e 2005, responsáveis por infecções hospitalares (14), infecções comunitárias (7) e colonizações (24), e 7 isoladas de estações de tratamento de esgoto (ETE) em 2009, em São Paulo, geneticamente distintas e produtoras de ESBL da família Enterobacteriaceae, foram estudadas. A técnica de PCR seguida de sequenciamento foi utilizada para a identificação dos genes blaESBL, triagem de elementos móveis e mapeamento do ambiente genético de blaESBL. A identificação dos grupos de incompatibilidade plasmidial (Inc) foi realizada pela técnica de PBRT, e a determinação dos tamanhos dos plasmídios pela técnica de S1-PFGE. Resultados. Os genes blaESBL identificados foram: amostras clínicas - blaTEM-15, blaTEM-197, blaSHV-5, blaSHV-12, blaSHV-27, blaSHV-28, blaSHV-45, blaSHV-55, blaSHV-110, blaCTX-M-2, blaCTX-M-59 e blaCTX-M-131; amostras ambientais blaSHV-28, blaCTX-M-15 e blaCTX-M-8. Os genes blaTEM- 15 e blaTEM-197 estavam associados aos elementos Tn2* e Tn3, respectivamente. Os genes blaSHV-5 e blaSHV-12 estavam associados à IS26, e não foi possível determinar o ambiente genético dos demais genes blaSHV. Os genes blaCTX-M-2, blaCTX-M-59 e blaCTXM- 131 estavam inseridos em integrons de classe 1 complexos, blaCTX-M-15 estava associado à ISEcp1 interrompida pela IS26, e blaCTX-M-8 estava associado à IS10, também interrompida pela IS26. Os principais grupos Inc detectados foram IncA/C (37 por cento ) e IncF (30,4 por cento ). Exceto por 7 cepas clínicas, todas apresentavam plasmídios de alto peso molecular, entre 48,5kb e 388kb. Conclusões. Este estudo detectou 15 genes blaESBL diferentes, dos quais dois são genes novos (blaTEM-197 e blaCTX-M-131) e três são inéditos no Brasil (blaTEM-15, blaSHV-55 e blaSHV-110). A maioria das cepas deste estudo possuía genes blaESBL associados a elementos mobilizáveis, bem como continham plasmídios de grupos Inc envolvidos na disseminação da resistência antimicrobiana. Além disso, carreavam plasmídios provavelmente conjugativos. Os resultados deste estudo mostram genes de resistência associados a elementos mobilizáveis em cepas contendo elementos transferíveis. As cepas foram isoladas tanto em uma instituição de saúde como nas ETEs da Grande São Paulo, mostrando o potencial de disseminação da resistência da clínica para o ambiente em nossa região. / Introduction. Bacterial resistance is facilitated by selective pressure of antimicrobial use in clinical and other activities, as agriculture and livestock, and can be spread to nature through the inadequate discharge of sewage or by the use of sludge in agriculture. Extended-spectrum -lactamases (ESBL) are the most prevalente forms of resistance in Gram-negative bacteria in the world, and their encoding genes are disseminated through several genetic elements, especially transposons and integrons mobilized to plasmids. Objective. To identify and characterize ESBL-encoding genes, as well as their probable mobilization pathways, in enterobacteria isolated from clinical and environmental sources. Material and Methods. Forty-five strains isolated from a public hospital in 2004 and 2005, responsible for hospital infections (14), community-acquired infections (7) and colonizations (24), and 7 isolated from sewage treatment plants (ETE) in 2009, in São Paulo, genetically distinct and ESBL producers from Enterobacteriaceae family, were studied. PCR technique followed by sequencing was used for blaESBL genes identification, mobile elements screening and blaESBL genetic environment mapping. Plasmid incompatibility groups (Inc) were identified by PBRT technique, and plasmid sizes were determined by S1-PFGE technique. Results. The blaESBL genes identified were: clinical samples - blaTEM-15, blaTEM-197, blaSHV-5, blaSHV-12, blaSHV-27, blaSHV-28, blaSHV-45, blaSHV-55, blaSHV-110, blaCTX-M-2, blaCTX-M-59 and blaCTX-M-131; environmental samples blaSHV-28, blaCTX-M-15 and blaCTX-M-8. Genes blaTEM-15 and blaTEM-197 were associated to the elements Tn2* and Tn3, respectivelly. Genes blaSHV-5 and blaSHV-12 were associated to IS26, and it was not possible to detect the genetic environment of the other blaSHV genes. Genes blaCTX-M-2, blaCTX-M-59 and blaCTX-M-131 were inserted in complex class 1 integrons, blaCTX-M-15 was associated to ISEcp1 interrupted by IS26, and blaCTX-M-8 was associated to IS10, also interrupted by IS26. The most common Inc grups detected were IncA/C (37 per cent ) and IncF (30,4 per cent ). Except for 7 clinical strains, all isolates showed high molecular weight plasmids, rangng from 48,5kb to 388kb. Conclusions. This study detected 15 different blaESBL genes, from which 2 are new genes (blaTEM-197 e blaCTX-M-131) and 3 are still unpublished in Brazil (blaTEM-15, blaSHV-55 and blaSHV-110). Most of the strains from this study had blaESBL genes associated to mobile elements, as well as they had plasmids from Inc groups involved in the spread of antimicrobial resistance. Moreover, the strains probably carried conjugative plasmids. Results from the present work show resistance genes associated to mobile elements in strains carrying transferable elements. The strains were isolated either from a healthcare institution or from ETEs in São Paulo, which shows the spread potential of resistance from the clinic to the environment in our region.

Salmonella é o agente etiológico mais comumente envolvido em casos e surtos de doenças diarréicas de origem alimentar no Brasil e outros países. A preocupação com este patógeno é, ainda, maior quando se verifica o surgimento e disseminação de cepas multi-resistentes e potencialmente mais patogênicas. O presente estudo teve como objetivo caracterizar 237 cepas Salmonella spp. distribuídas entre 50 sorovares diferentes, isoladas de alimentos associados e não associados à toxinfecções alimentares, quanto ao perfil de susceptibilidade antimicrobiana e presença dos genes de virulência spvC, invA, sefA e pefA. O gene invA foi detectado em todas as cepas de Salmonella. Com relação aos demais genes estudados, spvC e pefA foram encontrados em 48,1% e 44,3% das cepas, respectivamente. O gene sefA foi detectado em 31,6% das cepas, estando presente somente entre as cepas de S. Enteritidis. Ainda com relação à presença dos genes de virulência, as cepas de S. Enteritidis foram classificadas em três perfis, com predominância (90,7%) do perfil constituído pelos quatro genes de virulência. Quanto ao perfil de susceptibilidade antimicrobiana, 46,8% do total de cepas avaliadas foram sensíveis a todos os agentes antimicrobianos, 51,9% resistentes à pelo menos uma droga e 1,3% das cepas apresentaram apenas resistência intermediária. Multi-resistência foi observada em 10,5% das cepas. As maiores taxas de resistência foram observadas para estreptomicina (35,9%), ácido nalídixico (16,9%), tetraciclina (5,9%) e gentamicina (4,6%). Não foram detectadas cepas resistentes à cefoxitina, cefalotina, cefotaxima, amicacina, ciprofloxaxina e imipenem. Os resultados do presente estudo mostram a ampla distribuição dos genes de virulência e ocorrência de resistência antimicrobiana tanto nas cepas associadas a surtos como naquelas não envolvidas em toxinfecções alimentares, sendo os produtos de origem avícola fontes importantes de Salmonella com estas características. / Salmonella is the most common causative agent of cases and outbreaks of foodborne diarrhoeal diseases in Brazil and other countries. The concern with this pathogen is even greater considering the emergence and spread of multi-resistant and potentially more pathogenic strains. In this study, the antimicrobial susceptibility profile and the presence of virulence genes spvC, invA, sefA and pefA were examined in 237 Salmonella strains belonging to 50 serovars, isolated from foods associated and nonassociated to foodborne disease outbreaks. The gene invA was detected in all Salmonella strains. The genes spvC and pefA were found in 48.1% and 44.3% of strains, respectively. The sefA gene was found in 31.6% of strains and detected only in S. Enteritidis strains. According to the presence of virulence genes, S. Enteritidis strains were grouped in into three profiles, being the one consisting of four virulence genes the most common profile (90.7%). Among strains, 46.8% were sensitive to all antibiotics, 51.9% resistant to at least one drug and 1.3% of the strains presented intermediate resistance. Multi-resistance was seen in 10.5% of the strains. The highest rates of resistance were observed for streptomycin (35.9%), nalidixic acid (16.9%), tetracycline (5.9%) and gentamicin (4.6%). No resistance was observed to cefoxitin, cephalothin, cefotaxime, amikacin, ciprofloxaxina and imipenem. The results of this study show the wide distribution of virulence genes and occurrence of antimicrobial resistance in strains both associated and non-associated to foodborne disease outbreaks, being poultry products the major sources of Salmonella with these characteristics.

O presente estudo teve o objetivo de avaliar o perfil de sensibilidade in vitro de 171 isolados de Staphylococcus hyicus a nove antimicrobianos utilizados na suinocultura. A coleta das amostras foi realizada através de suabes de pele de leitões das fases de maternidade e creche provenientes de casos de epidermite exsudativa. As amostras foram submetidas ao isolamento e identificação e compreenderam dois grupos, classificadas de acordo com o período em que foram coletadas. O primeiro grupo foi formado por amostras históricas (80 isolados) provenientes de estudos anteriores entre 1975 a 1984 no Rio Grande do Sul, e mantidos liofilizados no laboratório de sanidade do Setor de Suínos (FAVET-UFRGS), e o segundo grupo (91 isolados) coletados no ano de 2012 nos estados do Rio Grande do Sul, Santa Catarina, Paraná e Minas Gerais. A realização do teste de susceptibilidade foi feita utilizando a técnica de diluição em ágar conforme protocolo M31-A3 do CLSI. Através de valores de Concentração Inibitória Mínima (CIM), foi obtido o perfil quantitativo de resistência em relação a nove antimicrobianos utilizados na suinocultura. A análise estatística possibilitou a avaliação de mudanças entre os dois grupos de amostras isoladas em diferentes épocas. Os resultados obtidos nesse estudo revelaram que para todas as cepas obtidas os beta-lactâmicos (ceftiofur e amoxicilina) foram os antimicrobianos mais ativos, com CIM para 90% dos isolados históricos (CIM90) de 0.5 μg/mL e 2.0 μg/mL para isolados recentes. A lincomicina, sulfametoxazol e tilmicosina foram os antimicrobianos menos ativos, com CIM90 variando de 128 a 256 μg/mL. O aumento na resistência entre o grupo de amostras históricas e recentes foi observado em seis dos nove antimicrobianos testados (amoxicilina, enrofloxacina, florfenicol, lincomicina, tilmicosina e tiamulina), em particular para a enrofloxacina e tiamulina. No caso do sulfametoxazole e tetraciclina, não foram observadas diferenças significativas na resistência, entretanto altos valores de CIM foram observados nos dois grupos avaliados (p<0,05). / This study was performed to evaluate the "in vitro" susceptibility profile of 171 Staphylococcus hyicus isolates from cases of exudative epidermitis to nine antibiotics used in pig production. Sample consisted of skin swabs from weanling and nursery piglets from farms suspected of exudative epidermitis. Samples were processed for bacterial isolation and identification and divided in two groups, according to the time of collection. The first group comprised historical samples (80 isolates) from previous studies (1975-1984) conducted in the state of Rio Grande do Sul, and stocked lyophilized in Swine Health Lab (FAVET-UFRGS), and the other group (91 isolates) formed by isolates collected in 2012 in the states of Rio Grande do Sul, Santa Catarina, Paraná and Minas Gerais, Brazil. The test was carried out using agar dilution protocol, according to M31-A3 CLSI. Through Minimal Inhibitory Concentrations (MIC) data, analysis was performed to get a quantitative profile of resistance to antimicrobials. Statistical analysis was performed to evaluate changes in resistance between the groups studied. Results showed that beta-lactam (ceftiofur and amoxicillin) were the most active antimicrobials for all strains tested, with the lowest MIC for inhibition of 90% of isolates (MIC90) of 0.5 μg/ml for historical isolates and of 2.0 μg/ml for recent isolates. Lincomycin, sulfamethoxazole and tilmicosin were the less active antimicrobials, with MIC90 varying from 128 to 256 μg/ml. Significant increase in resistance between historical and recent strains was observed for six out of nine antimicrobials tested (amoxicillin, enrofloxacin, florfenicol, lincomycin, tilmicosin and tiamulin), in particular for enrofloxacin e tiamulin. For sulfamethoxazole and tetracycline there was no significant differences in resistance between strains collected in different periods, however high MIC values were observed for these drugs.

Salmonella é o agente etiológico mais comumente envolvido em casos e surtos de doenças diarréicas de origem alimentar no Brasil e outros países. A preocupação com este patógeno é, ainda, maior quando se verifica o surgimento e disseminação de cepas multi-resistentes e potencialmente mais patogênicas. O presente estudo teve como objetivo caracterizar 237 cepas Salmonella spp. distribuídas entre 50 sorovares diferentes, isoladas de alimentos associados e não associados à toxinfecções alimentares, quanto ao perfil de susceptibilidade antimicrobiana e presença dos genes de virulência spvC, invA, sefA e pefA. O gene invA foi detectado em todas as cepas de Salmonella. Com relação aos demais genes estudados, spvC e pefA foram encontrados em 48,1% e 44,3% das cepas, respectivamente. O gene sefA foi detectado em 31,6% das cepas, estando presente somente entre as cepas de S. Enteritidis. Ainda com relação à presença dos genes de virulência, as cepas de S. Enteritidis foram classificadas em três perfis, com predominância (90,7%) do perfil constituído pelos quatro genes de virulência. Quanto ao perfil de susceptibilidade antimicrobiana, 46,8% do total de cepas avaliadas foram sensíveis a todos os agentes antimicrobianos, 51,9% resistentes à pelo menos uma droga e 1,3% das cepas apresentaram apenas resistência intermediária. Multi-resistência foi observada em 10,5% das cepas. As maiores taxas de resistência foram observadas para estreptomicina (35,9%), ácido nalídixico (16,9%), tetraciclina (5,9%) e gentamicina (4,6%). Não foram detectadas cepas resistentes à cefoxitina, cefalotina, cefotaxima, amicacina, ciprofloxaxina e imipenem. Os resultados do presente estudo mostram a ampla distribuição dos genes de virulência e ocorrência de resistência antimicrobiana tanto nas cepas associadas a surtos como naquelas não envolvidas em toxinfecções alimentares, sendo os produtos de origem avícola fontes importantes de Salmonella com estas características. / Salmonella is the most common causative agent of cases and outbreaks of foodborne diarrhoeal diseases in Brazil and other countries. The concern with this pathogen is even greater considering the emergence and spread of multi-resistant and potentially more pathogenic strains. In this study, the antimicrobial susceptibility profile and the presence of virulence genes spvC, invA, sefA and pefA were examined in 237 Salmonella strains belonging to 50 serovars, isolated from foods associated and nonassociated to foodborne disease outbreaks. The gene invA was detected in all Salmonella strains. The genes spvC and pefA were found in 48.1% and 44.3% of strains, respectively. The sefA gene was found in 31.6% of strains and detected only in S. Enteritidis strains. According to the presence of virulence genes, S. Enteritidis strains were grouped in into three profiles, being the one consisting of four virulence genes the most common profile (90.7%). Among strains, 46.8% were sensitive to all antibiotics, 51.9% resistant to at least one drug and 1.3% of the strains presented intermediate resistance. Multi-resistance was seen in 10.5% of the strains. The highest rates of resistance were observed for streptomycin (35.9%), nalidixic acid (16.9%), tetracycline (5.9%) and gentamicin (4.6%). No resistance was observed to cefoxitin, cephalothin, cefotaxime, amikacin, ciprofloxaxina and imipenem. The results of this study show the wide distribution of virulence genes and occurrence of antimicrobial resistance in strains both associated and non-associated to foodborne disease outbreaks, being poultry products the major sources of Salmonella with these characteristics.

Rising Antimicrobial Resistance is a threat faced all over the world. Bacterial infections that were treatable with antibiotics only a few years ago can now lead to life-threatening conditions. This thesis is part of the work of a large platform, PLATINEA, trying to reduce the rate of new resistances occurring in Sweden by preventing non optimal treatment. Due to shortages of the right antibiotics, suboptimal antibiotics are prescribed, which has shown to be accelerating the resistances among the bacterial populations. This study proposes an information exchange database and a central storage model for critical antibiotics to circumvent stock outs and inconveniences resulting from shortages of medically valuable antibiotics. Through interviewing prominent actors in the Swedish pharmaceutical supply chain an inside into the procurement of antibiotic in Sweden and what concerns are faced by the organs involved was created. Literature studies on occurred shortages of antibiotics in Sweden and the world were examined and possible reasons for these were identified. Examination of governmental efforts and assignments created the context in which gaps were identified that this thesis work could fill. A focus on Benzylpenicillin and Rifampicin were kept throughout the study. The collected data led to the implementation recommendation of two models by this study. An information platform suggested to allow better, faster and more accurate information exchange between all involved actors of the supply chain as well as a centralized storage model for the storage of antibiotics with medically high value in Sweden.  Through the implementation of the model systems shortages of critical antibiotics can be circumvented and better availability of information leads to quicker reactions ability to stock outs of other antibiotics.

L’antibiorésistance est une préoccupation majeure et globale de santé publique. La problématique de recherche de cette thèse visait à fournir au gestionnaire une aide à la décision pour la mise en œuvre et l’évaluation des mesures de lutte contre l’antibiorésistance en santé animale. Pour espérer limiter le phénomène et le gérer, il faut en connaître la dynamique d’évolution : la surveillance des résistances se positionne donc comme un enjeu clé de la lutte. En premier lieu, ces travaux ont déterminé, au travers d’une enquête populationnelle et d’une approche sociologique, dans quelles mesures les données collectées par le réseau d’épidémiosurveillance de l’antibiorésistance des bactéries pathogènes animales (Résapath) pouvaient être utilisées pour répondre à la problématique. La représentativité et la couverture du Résapath estimées satisfaisantes, l’évolution des résistances a été investiguée avec des modèles additifs généralisés à partir des données de surveillance. La mise en parallèle des tendances des résistances avec les mesures de maîtrise a illustré l’impact positif des changements de pratique sur l’évolution des résistances. Enfin, dans la perspective du concept One Health, qui prône une approche intégrée et collaborative de la santé, le rapprochement entre les dynamiques des résistances chez les animaux et chez l’Homme a été exploré, en se basant sur les données du réseau MedQual en médecine de ville. Les tendances des résistances ont évolué de manière spécifique à chaque espèce, sans dynamique commune entre l’Homme et les animaux. Les efforts pour lutter contre l’antibiorésistance doivent donc être menés de concert, tant en médecine humaine que vétérinaire / Antimicrobial resistance is a major and global public health concern. In this context, the research problem was to provide decision support to the risk manager for the implementation and assessment of control measures in animal health. To limit and manage the phenomenon, we must know the dynamic of its evolution: the surveillance is therefore a key element in the fight against antimicrobial resistance. At first, using a population survey and a sociological approach, this research determined to what extent the data collected by the French surveillance network of antimicrobial resistance in diseased animals (RESAPATH) could be used to answer the research problem. Since the representativeness and the coverage of the RESAPATH were considered satisfactory, surveillance data were used to characterize the dynamics of the resistances and generalized additive models were developed. The comparison of resistance trends and control measures underscored the positive impact of changes in practices on the evolution of resistances. Finally, in the context of the ‘One Health’ concept that advocates an integrated and collaborative approach to health, the parallel was drawn between resistances in isolates from animals and humans. Data from the French surveillance network of antimicrobial resistance of bacteria isolated in community (MedQual) were analysed. Resistance dynamics were specific to each species. These results advocate that the efforts to fight antimicrobial resistance must be carried in all sectors and for all species, both in human and veterinary medicine

<p>The first hospital outbreak of vancomycin-resistant enterococci (VRE) and carriage rates of VRE and ampicillin-resistant enterococci (ARE) in Sweden were investigated. Clonal relationships and mutations in fluoroquinolone resistance determining regions among ARE collected nation-wide were studied. Risk factors for ARE infection, shedding of ARE and the presence of the virulence gene <i>esp</i> in ARE isolates and patients on a hematology unit and other units at Uppsala University Hospital were further investigated. </p><p>The first Swedish hospital VRE outbreak was due to clonal spread of <i>E. faecium, vanA</i>. The nation wide carriage rates of ARE and VRE were 21.5% / 1% and 6% / 0%, among hospitalized patients and non-hospitalized individuals respectively. All ARE and VRE were <i>E. faecium</i> and >90% resistant to ciprofloxacin. All VRE carried<i> vanB</i>. Carriage of ARE was independently associated with >5 days of antibiotic treatment. Phenotypic and genetic typing showed a significantly higher homogeneity among ARE compared to matched ASE <i>E. faecium</i> isolates. Mutations conferring high-level ciprofloxacin resistance were found only in ARE. Risk factors for ARE infection included long duration of hospital stay and exposure to antibiotics. Skin carriage was associated with ARE shedding. ARE bacteremia was independently associated with prior ARE colonization and hematopoietic stem cell transplantation. Death was more common in ARE septicemia cases compared to controls. <i>Esp</i> was significantly more common in ARE surveillance compared to ARE blood isolates from patients on the hematology ward.</p><p>In conclusion, VRE were rare but clonally related multi-resistant ARE <i>E. faecium</i> were highly prevalent in Swedish hospitals. Spread of ARE in hospitals during the 1990s is suggested to be the main explanation for the emergence of ARE in Sweden. Spread was facilitated by use of antibiotics and probably by the presence of virulence genes in<i> E. faecium</i> isolates.</p>