ESBL in positive hemoculture of a Southern-Brazil Teaching Hospital's Intensive Care Units / ESBL em hemoculturas positivas das Unidades de Terapia Intensiva de um Hospital Universitário do sul do Brasil

Bloodstream infections are among the most serious and frequent infections, and the people most exposed are patients in the Intensive Care Unit (ICU). ESBL (extended-spectrum beta-lactate) are resistant bacteria to penicillins, cephalosporins and monobactams. It´s necessary to know how often and which microorganisms are involved, checking their susceptibility. This study was carried out at the University Hospital. Data collection was performed in the Adult and Newborn ICUs, with assessment of microorganisms and their resistance profile. During six-month period, 156 samples were studied, and 42 were positive with microorganism isolation. Isolated species include Staphylococcus aureus, Staphylococcus epidermidis and Klebsiella pneumoniae. Many resistant to carbapenem.

ESBL in Positive Hemoculture of a Southern-Brazil Teaching Hospital's Intensive Care Units As infecções da corrente sanguínea estão entre as infecções mais graves e frequentes, e os indivíduos mais expostos são os pacientes da Unidade de Terapia Intensiva (UTI). As ESBL (Beta-Lactamase de Espectro Estendido) são bactérias resistentes a penicilinas, cefalosporinas e monobactâmicos. Se faz necessário saber com que frequência e quais microrganismos estão envolvidos, verificando sua suscetibilidade. Este estudo foi realizado no Hospital Universitário. A coleta de dados foi realizada nas UTIs Adulto e Neonatal, com avaliação dos microrganismos e seu perfil de resistência. Durante o período de seis meses, foram estudadas 156 amostras, sendo 42 positivas com isolamento dos microrganismos. As espécies isoladas incluem Staphylococcus aureus, Staphylococcus epidermidis e Klebsiella pneumoniae. Muitos resistentes aos carbapenêmicos.

Detection of beta-lactamase-producing Enterobacteriaceae in a veterinary hospital environment / Detecção de Enterobactérias produtoras de beta-lactamases em ambiente hospitalar veterinário

Due to the strong selective pressure resulting from the misuse of antibiotics, the natural process of bacterial resistance has been accelerated, leading to the increasingly constant appearance of multiresistant isolates. The high number of multi-resistant bacteria is a one health problem. Enterobacteriaceae are usually commensal bacteria of the gastrointestinal tract. However, they can cause infections, and the most important resistance characteristic among them is the production of ß-lactamases. This study aimed to identify ESBL-producing Enterobacteriaceae of types of TEM, SHV, and the CTX-Mgroups. To isolate the enterobacteria, swabs were collected by swiping objects that had contact with the patients and professionals, and the water of the hospital environment. Ten collections were carried out, yielding 306 samples, from which 118 enterobacteria were identified: Escherichia coli, Enterobacter spp., Klebsiella spp., Proteus mirabilis, Serratiaspp., and Citrobacter spp. Isolates. The genes TEM and CTX-M, for the production of ß-lactamases, were detected in 12.7% of the 118 enterobacterial isolates. It is very important to know the bacterial population circulating in the veterinary hospital environment and its resistance to antimicrobials so that professionals can take appropriate measures to minimize the risks of transmission, especially from cages and consultation tables. In addition, the correct control of the microbiological quality of the supply water, as well as environmental cleaning procedures, are essential to prevent the transmission of these microorganisms.(AU)

Devido à grande pressão seletiva decorrente do uso indevido de antibióticos, tem se acelerado o processo natural de resistência das bactérias, levando ao aparecimento cada vez mais constante de isolados multirresistentes. O elevado número de bactérias multirresistentes identificadas é um problema da saúde única. As enterobactérias são bactérias geralmente comensais do trato gastrointestinal, entretanto podem causar infecções, e a característica de resistência mais importante entre elas é a produção de ß-lactamases. Buscando caracterizar melhor os microrganismos circulantes e potencialmente causadores de infecções em ambiente hospitalar veterinário, este estudo objetivou identificar as enterobactérias produtoras de ESBL do tipo TEM, SHV e os cinco grupos de CTX-M presentes em isolados circulantes em hospital veterinário. Foi realizada coleta de suabes de arrasto de objetos que entram em contato com os pacientes e com os profissionais que ali trabalham, bem como de água, para a identificação das enterobactérias. Foram realizadas 10 coletas, obtendo-se 306 amostras, dessas, 118 enterobactérias foram identificadas: Escherichia coli, Enterobacter, Klebsiella, Proteus mirabilis, Serratia e Citrobacter. Dentre as enterobactérias identificadas, alguns isolados possuíam genes para a produção de ß-lactamases, do tipo TEM e CTX-M. É de grande importância conhecer a população bacteriana circulante no ambiente hospitalar veterinário, e a sua resistência aos antimicrobianos, para que os profissionais possam tomar medidas apropriadas para minimizar os riscos de transmissão, principalmente a partir de gaiolas e mesas de atendimento. Além disso, o correto controle da qualidade microbiológica da água de abastecimento, bem como dos procedimentos de higienização do ambiente, são fundamentais para evitar a transmissão destes microrganismos.(AU)

Acinetobacter calcoaceticus-Acinetobacter baumannii complex in animals: identification and antimicrobial resistance profile / Complexo Acinetobacter calcoaceticus-Acinetobacter baumannii em animais: identificação e perfil de resistência antimicrobiana

Acinetobacter spp. is emerging as an important human and veterinary pathogen, mostly due to intrinsic and acquired resistance to antimicrobials. Despite its public health relevance, little is known about the prevalence, role of different Acinetobacter species and antimicrobial resistance profile of animal-origin isolates. Traditional phenotypic tests may fail to discriminate Acinetobacter species, therefore molecular analyses are often required as a complementary approach. The objectives of this study were to evaluate the occurrence of strains of the Acinetobacter calcoaceticus-Acinetobacter baumannii (Acb) complex isolated from animal infections including urinary tract infections, otitis, piodermitis and pododermatitis, and its resistance profile against different antimicrobial classes, including carbapenems. All Gram-negative coccobacilli isolates were characterized by MALDI-TOF and multiplex PCR, and the disk diffusion test was used to investigate multi-drug resistance (MDR) and carbapenem resistance genes by PCR as preconized by the standard guidelines. MALDI-TOF technique identified 21 strains belonging to the Acb complex (10 A. pittii, 8 A. baumannii, 3 A. nosocomialis, 1 A. ursingii, and 1 A. venetianus). Multiplex PCR confirmed the results of MALDI-TOF for 20 strains. Eight strains (34.78%) were classified as MDR, being 50% (4/8) A. baumannii, 37.5% (3/8) A. pittii, and 12.5% (1/8) A. nosocomialis. None of the isolates presented phenotypic carbapenemase production. Considering the carbapenem resistance genes, 26.09% (6/23) of the isolates presented one or more carbapenemase genes. From these, 50% (3/6) presented only bla VIM, 33.33% (2/6) presented only blaIMP, and 16.67% (1/6) presented blaIMP e blaVIM, simultaneously. These genes were detected among A. pittii isolates mostly (66.67%, 4/6). This study provides further insights into the occurrence and resistance profile of Acinetobacter of animal origin.

Acinetobacter spp. está emergindo como um importante patógeno humano e veterinário, principalmente devido à resistência intrínseca e adquirida aos antimicrobianos. Apesar de sua relevância para a saúde pública, pouco se sabe sobre a prevalência, o papel das diferentes espécies de Acinetobacter e o perfil de resistência antimicrobiana de isolados de origem animal. Testes fenotípicos tradicionais podem falhar em discriminar espécies de Acinetobacter, portanto, análises moleculares são frequentemente necessárias como uma abordagem complementar. Os objetivos deste estudo foram avaliar a ocorrência de cepas do complexo Acinetobacter calcoaceticus-Acinetobacter baumannii (Acb) isolados de infecções de animais, incluindo infecções do trato urinário, otite, piodermite e pododermatite, e seu perfil de resistência a diferentes classes de antimicrobianos, incluindo os carbapenêmicos. Todas as cepas cocobacilos Gram-negativas foram caracterizados por MALDI-TOF e PCR multiplex, e o teste de difusão em disco foi usado para investigar genes de resistência a múltiplas drogas (MDR) e resistência a carbapenêmicos por PCR conforme preconizado pelas diretrizes padrão. A técnica MALDI-TOF identificou 21 cepas pertencentes ao complexo Acb (10 A. pittii, 8 A. baumannii, 3 A. nosocomialis, 1 A. ursingii e 1 A. venetianus). Multiplex PCR confirmou os resultados de MALDI-TOF para 20 cepas. Oito cepas (34.78%) foram classificadas como MDR, sendo 50% (4/8) A. baumannii, 37.5% (3/8) A. pittii e 12.5% (1/8) A. nosocomialis. Nenhum dos isolados apresentou produção fenotípica de carbapenemases. Considerando os genes de resistência a carbapenemas, 26.09% (6/23) dos isolados apresentaram um ou mais genes de carbapenemases. Destes, 50% (3/6) apresentaram apenas bla VIM, 33.33% (2/6) apresentaram apenas bla IMP e 16.67% (1/6) apresentaram bla IMP e bla VIM, simultaneamente. Esses genes foram detectados principalmente entre os isolados de A. pittii (66.67%, 4/6). Este estudo fornece mais informações sobre a ocorrência e perfil de resistência de Acinetobacter de origem animal.

Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil / Detecção dos principais microrganismos multirresistentes no ambiente de um hospital veterinário de ensino no Brasil

Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.(AU)

A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.(AU)

Detection of the main multiresistant microorganisms in the environment of a teaching veterinary hospital in Brazil / Detecção dos principais microrganismos multirresistentes no ambiente de um hospital veterinário de ensino no Brasil

Contamination of the veterinary hospital environment with multiresistant pathogens endangers not only hospitalized animals, but also the workplace safety of veterinarians and nurses, animal guardians and, when in case of a teaching hospital, veterinary students. The objective of this study was to map the main points of bacterial contamination of a veterinary teaching hospital in Brazil to identify multiresistant microorganisms and their antimicrobial resistance genes. Samples were collected from 39 different locations of a veterinary school hospital which comprised a pool according to each hospital environment. In certain environments, more than one pool has been collected. All samples were collected in quadruplicates for the selective isolation of the main multiresistant microorganisms: methicillin-resistant Staphylococcus (MRS), vancomycin resistant Enterococcus (VRE), cephalosporinases and/or extended-spectrum beta-lactamase-producing Gram-negative bacteria (ESBL) and Carbapenemase-producing (CP). After isolation and identification of isolates, multiplex-PCR reactions were performed to detect the main genes for each microorganism and antimicrobial susceptibility tests with the main antibiotics used for each bacterial group according to CLSI. Of the 39 veterinary teaching hospital sites collected, all (100%) had at least one of the microorganisms surveyed, and 17.95% (n=7) of the sites were able to isolate the four pathogens. From the 94 pools collected, it was possible to isolate MRS in 81.91% (n=77), VRE in 12.77% (n=12), cephalosporinases and/or ESBL in 62.77% (n=59) and CP in 24.47%. (n=23). Regarding MRS, the mecA gene was detected in all isolates. All isolated VREs were identified as Enterococcus faecalis and presented the vanA gene. Regarding cephalosporinases and/or ESBL, 89.83% (n=53) of the isolates presented the blaTEM gene, 57.63% (n=34) the blaOXA-1 gene, 37.29% (n=22) blaCTX-M gene from some group (1, 2, 9 ou 8/25) and 20.34% (n=12) the blaSHV gene. It was possible to identify the main microorganisms responsible for causing nosocomial infections in humans (VRE, MRS, ESBL and CP) in the veterinary hospital environment, suggesting a source of infection for professionals and students of veterinary medicine, placing a high risk for public health.(AU)

A contaminação do ambiente hospitalar veterinário com patógenos multirresistentes coloca em perigo não apenas os animais hospitalizados, mas também a segurança no local de trabalho de veterinários e enfermeiros, responsáveis por animais e, quando se tratar de um hospital de ensino, estudantes de veterinária. O objetivo deste estudo foi mapear os principais pontos de contaminação bacteriana de um hospital veterinário de ensino no Brasil, identificando microorganismos multirresistentes e seus genes de resistência antimicrobiana. As amostras foram coletadas em 39 locais diferentes de um hospital de escola veterinária, que compreendia um pool de acordo com o ambiente de cada hospital. Em certos ambientes, mais de um pool foi coletado. Todas as amostras foram coletadas em quadruplicados para o isolamento seletivo dos principais microorganismos multirresistentes: Staphylococcus resistente à meticilina (MRS), Enterococcus resistente à vancomicina (VRE), bactérias Gram-negativas produtoras de cefalosporinases e/ou beta-lactamase de espectro estendido (ESBL) e produtoras de carbapenemase (PC). Após o isolamento e identificação dos isolados, foram realizadas reações de PCR multiplex para detectar os principais genes de cada microorganismo e testes de susceptibilidade a antimicrobianos com os principais antibióticos utilizados para cada grupo bacteriano de acordo com o CLSI. Dos 39 locais do VCH coletados, todos (100%) possuíam pelo menos um dos microrganismos pesquisados e 17,95% (n=7) dos locais foram capazes de isolar os quatro patógenos. Dos 94 pools coletados, foi possível isolar MRS em 81,91% (n=77), VRE em 12,77% (n=12), ESBL em 62,77% (n=59) e carbapenemases em 24,47% (n=23). Em relação ao MRS, o gene mecA foi detectado em todos os isolados. Todos os VREs isolados foram identificados como Enterococcus faecalis e apresentaram o gene vanA. Em relação às cefalosporinases e/ou ESBL, 89,83% (n=53) dos isolados apresentaram o gene blaTEM, 57,63% (n=34) o gene blaOXA-1, 37,29% (n=22) o gene blaCTX-M de algum grupo e 20,34% (n=12) o gene blaSHV. Foi possível identificar os principais microrganismos responsáveis por causar infecções nosocomiais em humanos (VRE, MRS, ESBL e CP) no ambiente hospitalar veterinário, sugerindo uma fonte de infecção para profissionais e estudantes de medicina veterinária, colocando alto risco para a saúde pública.(AU)

Detecção de beta-lactamase por Staphylococcus coagulase-negativo isolados de queijo mussarela fatiados e fatiadores de frios / Beta-lactamase detection in coagulase-negative Staphylococci isolated from sliced mozzarella cheese and cold-cut slicers

A resistência de bactérias a antimicrobianos é considerada um problema de saúde pública, sendo a resistência aos beta-lactâmicos uma das mais importantes. O objetivo do presente estudo foi detectar a produção da enzima β-lactamase por isolados de Staphylococcus coagulase-negativo (SCN), provenientes de queijos Mussarela fatiados e equipamentos de fatiamento de frios. Os testes foram realizados utilizando discos impregnados com cefalosporina cromógena para detecção da β-lactamase. Dos 103 isolados de Staphylococcus spp. analisados, 55 (53%) produziram β-lactamase e 48 (47%) não produziram. Portanto, é possível inferir que SCN isolados neste estudo, podem inativar antimicrobianos β-lactâmicos e assim, exercer influência negativa na saúde pública, devido ao potencial em transferir genes de resistência antimicrobiana para outras bactérias.(AU)

Detecção de beta-lactamase por Staphylococcus coagulase-negativo isolados de queijo mussarela fatiados e fatiadores de frios / Beta-lactamase detection in coagulase-negative Staphylococci isolated from sliced mozzarella cheese and cold-cut slicers

A resistência de bactérias a antimicrobianos é considerada um problema de saúde pública, sendo a resistência aos beta-lactâmicos uma das mais importantes. O objetivo do presente estudo foi detectar a produção da enzima β-lactamase por isolados de Staphylococcus coagulase-negativo (SCN), provenientes de queijos Mussarela fatiados e equipamentos de fatiamento de frios. Os testes foram realizados utilizando discos impregnados com cefalosporina cromógena para detecção da β-lactamase. Dos 103 isolados de Staphylococcus spp. analisados, 55 (53%) produziram β-lactamase e 48 (47%) não produziram. Portanto, é possível inferir que SCN isolados neste estudo, podem inativar antimicrobianos β-lactâmicos e assim, exercer influência negativa na saúde pública, devido ao potencial em transferir genes de resistência antimicrobiana para outras bactérias.

Extended Spectrum Beta-Lactamase Production and Biofilm Formation in Salmonella Serovars Resistant to Antimicrobial Agents

Antimicrobial resistance is a serious public health problem and Salmonella spp. is highly resistant to antimicrobial agents. Biofilms are important in the food industry due to their formation on products, utensils, and surfaces and the difficulty in their removal. The objective of this study was to assess extended spectrum beta-lactamase (ESBL) production, antimicrobial resistance, and biofilm production of Salmonella isolated from poultry slaughterhouses. Antimicrobial susceptibility was assessed by the disk diffusion assay and ESBL by double diffusion disk assay using the beta-lactamase inhibitor (amoxicillin+clavulanate). The antimicrobials tested were: ampicillin, amoxicillin+clavulanate, aztreonam, ceftazidime, cefotaxime, chloramphenicol, gentamicin, enrofloxacin, sulfonamide, and tetracycline. Serovars Infantis, Panamá, and Tennessee were found to produce ESBL. All serovars were sensitive to tetracycline, and S. Brandenburg was sensitive to all drugs tested. Serovars Panamá, Anatum, Infantis, and Schwarzengrund were moderate biofilm producers at 3 ºC and 9 ºC±1 ºC, respectively, showing possible adaptation of these serovars to these temperatures. Antimicrobials should be used with caution because of the levels of resistance observed and because of ESBL production, and hygiene and sanitary measures should be enhanced to minimize the adhesion of biofilm-forming Salmonella serovars at refrigeration temperatures.(AU)

Antimicrobial resistance and extended-spectrum beta-lactamase production in Enterobacteriaceae isolates from household cats (Felis silvestris catus)

Background: In Brazil, cats in households has recently increased dramatically, likely due to their lower space and carerequirements. We need to know the health of these companion animal species, since they have behavioral patterns thatmake them an important link in the epidemiological chain. Extended spectrum beta-lactamase producer strains (ESBL)are resistant to penicillin, cephalosporin and monobactam, but they are susceptible to clavulanate. The goal of this study isto detect Enterobacteriaceae that produce extended spectrum beta-lactamase (ESBL) and evaluate the bacterial resistanceprofile in isolated cats (Felis silvestris catus) that live in a city located at west of Parana state, Brazil.Materials, Methods & Results: Swabs were aseptically collected from the anal orifice and oral cavity of 49 female domestic cats that were healthy upon clinical and physical examination, a minimum age of one year, weighing up to 3 kg,and had attended a veterinary clinic specializing in cats, in order to, later, perform the isolation and bacterial identification, antimicrobial sensibility phenotypic test and the phenotypic test to detect ESBL producer strains. From the 98 swabscollected it was possible to perform the bacterial isolation in 68 samples; 40.81% isolated from anal orifice and 28.57%isolated from oral cavity. From rectal and oral cavities 77.50% and 71.42% of the isolated were identified as Escherichiacoli respectively, being 2.94% considered ESBL producer strains. In relation to bacterial resistance the antibiotics thatshown more resistance in anal orifice were ampicillin, amoxicillin, nalidixic acid, sulfazotrim, tetracycline and aztreonam.In oral cavity they were ampicillin, amoxicillin, cefoxitin, amoxicillin + clavulanate, aztreonam, ceftriaxone and nalidixicacid; and the bacterial resistance index shown that 39.70% were considered high level risk...

Antimicrobial resistance and extended-spectrum beta-lactamase production in Enterobacteriaceae isolates from household cats (Felis silvestris catus)

Background: In Brazil, cats in households has recently increased dramatically, likely due to their lower space and carerequirements. We need to know the health of these companion animal species, since they have behavioral patterns thatmake them an important link in the epidemiological chain. Extended spectrum beta-lactamase producer strains (ESBL)are resistant to penicillin, cephalosporin and monobactam, but they are susceptible to clavulanate. The goal of this study isto detect Enterobacteriaceae that produce extended spectrum beta-lactamase (ESBL) and evaluate the bacterial resistanceprofile in isolated cats (Felis silvestris catus) that live in a city located at west of Parana state, Brazil.Materials, Methods & Results: Swabs were aseptically collected from the anal orifice and oral cavity of 49 female domestic cats that were healthy upon clinical and physical examination, a minimum age of one year, weighing up to 3 kg,and had attended a veterinary clinic specializing in cats, in order to, later, perform the isolation and bacterial identification, antimicrobial sensibility phenotypic test and the phenotypic test to detect ESBL producer strains. From the 98 swabscollected it was possible to perform the bacterial isolation in 68 samples; 40.81% isolated from anal orifice and 28.57%isolated from oral cavity. From rectal and oral cavities 77.50% and 71.42% of the isolated were identified as Escherichiacoli respectively, being 2.94% considered ESBL producer strains. In relation to bacterial resistance the antibiotics thatshown more resistance in anal orifice were ampicillin, amoxicillin, nalidixic acid, sulfazotrim, tetracycline and aztreonam.In oral cavity they were ampicillin, amoxicillin, cefoxitin, amoxicillin + clavulanate, aztreonam, ceftriaxone and nalidixicacid; and the bacterial resistance index shown that 39.70% were considered high level risk...(AU)

Extended Spectrum Beta-Lactamase Production and Biofilm Formation in Salmonella Serovars Resistant to Antimicrobial Agents

Antimicrobial resistance is a serious public health problem and Salmonella spp. is highly resistant to antimicrobial agents. Biofilms are important in the food industry due to their formation on products, utensils, and surfaces and the difficulty in their removal. The objective of this study was to assess extended spectrum beta-lactamase (ESBL) production, antimicrobial resistance, and biofilm production of Salmonella isolated from poultry slaughterhouses. Antimicrobial susceptibility was assessed by the disk diffusion assay and ESBL by double diffusion disk assay using the beta-lactamase inhibitor (amoxicillin+clavulanate). The antimicrobials tested were: ampicillin, amoxicillin+clavulanate, aztreonam, ceftazidime, cefotaxime, chloramphenicol, gentamicin, enrofloxacin, sulfonamide, and tetracycline. Serovars Infantis, Panamá, and Tennessee were found to produce ESBL. All serovars were sensitive to tetracycline, and S. Brandenburg was sensitive to all drugs tested. Serovars Panamá, Anatum, Infantis, and Schwarzengrund were moderate biofilm producers at 3 ºC and 9 ºC±1 ºC, respectively, showing possible adaptation of these serovars to these temperatures. Antimicrobials should be used with caution because of the levels of resistance observed and because of ESBL production, and hygiene and sanitary measures should be enhanced to minimize the adhesion of biofilm-forming Salmonella serovars at refrigeration temperatures.

Multidrug resistant and ESBL-producing Salmonella spp. isolated from poultry / Salmonella spp. produtora de ESBL e multirresistente a drogas isolada de frangos

Salmonella spp. is one of the main agents responsible for foodborne infection in humans, and products of poultry origin are the most common infection sources. Studies have shown the occurrence of antimicrobials resistant Salmonella spp. in animal products. The Extended Spectrum β-Lactamase (ESBL) are enzymes that confer to bacteria the ability to hydrolyze cephalosporin with an oximino side chain and monobactams. This study aimed to investigate antimicrobial resistance profile, identify phenotypes and genotypes for multiple drug resistance (MDR) and that produce ESBL from isolates of Salmonella spp. in the broiler production chain. We used samples of Salmonella spp. (n=11) isolates from poultry, poultry products and poultry-source environment from the state of Maranhão - Brazil. The isolates of Salmonella spp. assessed showed genotypical and phenotypical characteristics of MDR. The results show that 72.72% (08/11) of the strains presented the phenotypic profile for ESBL production. The isolates showed positivity to at least 13.64% (03/22) of the genes studied and the highest frequencies were observed in genes sul1 (73%), dfrA12 (55%), blaCTX-M (55%), tetA, tetB and tetC, with 45%. In conclusion, the strains of Salmonella spp. isolates present genotypic and phenotypic characteristics for MDR and ESBL production, demonstrating the dissemination risk of these microorganisms through the food chain.

Salmonella spp. é um dos principais agentes responsáveis por infecção de origem alimentar em humanos, sendo os produtos de origem avícola apontados como as fontes de infecção mais frequentes. Estudos demonstraram a ocorrência de Salmonella spp. resistente a antimicrobianos em produtos de origem animal. As β-lactamases de espectro extendido (ESBL) são enzimas que conferem às bactérias a capacidade de hidrolisar cefalosporinas com uma cadeia lateral oximino e monobactâmicos. Este estudo objetivou investigar o perfil de resistência a antimicrobianos, identificar fenótipos e genótipos de multirresistência a drogas (MDR), e produção de ESBL em isolados de Salmonella spp. provenientes da cadeia produtiva de frangos de corte. Foram utilizadas amostras de Salmonella spp. (n=11) isoladas de aves, produtos e ambiente de origem avícola do estado do Maranhão - Brasil. Os isolados de Salmonella spp. avaliados apresentaram resistência múltipla a drogas tanto genotípica quanto fenotipicamente. Os resultados mostram que 72,72% (08/11) das cepas apresentaram perfil fenotípico de produção de ESBL. Os isolados apresentaram positividade a pelo menos 13,64% (03/22) dos genes pesquisados, sendo as maiores frequências observadas nos genes sul1 (73%), dfrA12 (55%), blaCTX-M (55%), tetA, tetB e tetC, com 45%. Conclui-se que as cepas de Salmonella spp. isoladas apresentam características genotípicas e fenotípicas de multirresistência a drogas e produção de ESBL, demonstrando o risco de disseminação destes microrganismos ao longo da cadeia alimentar.

Multidrug resistant and ESBL-producing Salmonella spp. isolated from poultry / Salmonella spp. produtora de ESBL e multirresistente a drogas isolada de frangos

Salmonella spp. is one of the main agents responsible for foodborne infection in humans, and products of poultry origin are the most common infection sources. Studies have shown the occurrence of antimicrobials resistant Salmonella spp. in animal products. The Extended Spectrum β-Lactamase (ESBL) are enzymes that confer to bacteria the ability to hydrolyze cephalosporin with an oximino side chain and monobactams. This study aimed to investigate antimicrobial resistance profile, identify phenotypes and genotypes for multiple drug resistance (MDR) and that produce ESBL from isolates of Salmonella spp. in the broiler production chain. We used samples of Salmonella spp. (n=11) isolates from poultry, poultry products and poultry-source environment from the state of Maranhão - Brazil. The isolates of Salmonella spp. assessed showed genotypical and phenotypical characteristics of MDR. The results show that 72.72% (08/11) of the strains presented the phenotypic profile for ESBL production. The isolates showed positivity to at least 13.64% (03/22) of the genes studied and the highest frequencies were observed in genes sul1 (73%), dfrA12 (55%), blaCTX-M (55%), tetA, tetB and tetC, with 45%. In conclusion, the strains of Salmonella spp. isolates present genotypic and phenotypic characteristics for MDR and ESBL production, demonstrating the dissemination risk of these microorganisms through the food chain.(AU)

Salmonella spp. é um dos principais agentes responsáveis por infecção de origem alimentar em humanos, sendo os produtos de origem avícola apontados como as fontes de infecção mais frequentes. Estudos demonstraram a ocorrência de Salmonella spp. resistente a antimicrobianos em produtos de origem animal. As β-lactamases de espectro extendido (ESBL) são enzimas que conferem às bactérias a capacidade de hidrolisar cefalosporinas com uma cadeia lateral oximino e monobactâmicos. Este estudo objetivou investigar o perfil de resistência a antimicrobianos, identificar fenótipos e genótipos de multirresistência a drogas (MDR), e produção de ESBL em isolados de Salmonella spp. provenientes da cadeia produtiva de frangos de corte. Foram utilizadas amostras de Salmonella spp. (n=11) isoladas de aves, produtos e ambiente de origem avícola do estado do Maranhão - Brasil. Os isolados de Salmonella spp. avaliados apresentaram resistência múltipla a drogas tanto genotípica quanto fenotipicamente. Os resultados mostram que 72,72% (08/11) das cepas apresentaram perfil fenotípico de produção de ESBL. Os isolados apresentaram positividade a pelo menos 13,64% (03/22) dos genes pesquisados, sendo as maiores frequências observadas nos genes sul1 (73%), dfrA12 (55%), blaCTX-M (55%), tetA, tetB e tetC, com 45%. Conclui-se que as cepas de Salmonella spp. isoladas apresentam características genotípicas e fenotípicas de multirresistência a drogas e produção de ESBL, demonstrando o risco de disseminação destes microrganismos ao longo da cadeia alimentar.(AU)

Susceptibilidade de Staphylococcus aureus isolados de leite cru a antibióticos comerciais / Susceptibility of Staphylococcus aureus isolated from raw milk to commercial antibiotics

O leite possui extremo valor na dieta humana e constitui um excelente substrato para o crescimento de grande diversidade de microrganismos, dentre eles o Staphylococcus aureus, frequentemente encontrado no leite cru, e que se destaca como um dos microrganismos mais prevalentes em casos de mastite bovina no mundo. Objetivou-se avaliar a sensibilidade de cepas de S. aureus isoladas de leite cru à antimicrobianos comerciais. Para isso, selecionaram-se antibióticos com base em pesquisas junto a estabelecimentos comerciais de medicamentos veterinários no município de Zé Doca, Estado do Maranhão, Brasil. Esses antibióticos foram identificados por associação à base de penicilina, estreptomicina, isoniazida e prednisolona (BPC), oxitetraciclina base (TOR), associação à base de penicilina e estreptomicina (MPP) e cloridrato de tetraciclina (TCA). Posteriormente, realizou-se contagem de Staphylococcus spp., com isolamento e identificação bioquímica das cepas de S. aureus as quais, em seguida, foram submetidas à antibiogramas com identificação de ce,pas produtoras de beta-lactamase. Os resultados demonstraram contaminação em todas as amostras por Staphylococcus spp., com contagens de 1,9 x 103 a 5,24 x 106 UFC/mL. Dentre os antibióticos testados, o TOR apresentou maior eficiência na eliminação ou redução de todas as cepas de S. aureus. Verificou-se que 50% das cepas de S. aureus isoladas produziram enzima beta-lactamase. Torna-se necessário a criação de medidas de combate ao surgimento de novas cepas bacterianas e atuação das autoridades públicas, fiscalizando a comercialização de medicamentos veterinários.

Milk is extremelly valuable to the human diet and an excellent substrate for the growth of a wide range of microorganisms, including Staphylococcus aureus, frequently found in raw milk and one of the most prevalent microorganisms in cases of bovine mastitis in the world. The objective of this study was to evaluate the sensitivity of S. aureus strains isolated from raw milk to commercial antimicrobials. For this end, antibiotics based on research were selected from commercial establishments of veterinary drugs in the municipality of Zé Doca (Maranhão State, Brazil). The antibiotics were identified by association with penicillin, streptomycin, isoniazid and prednisolone (PCB), oxytetracycline base (TOR), a combination of penicillin and streptomycin (MPP), and tetracycline hydrochloride (TCA). Subsequently, Staphylococcus spp. counting was perfomed, with isolation and biochemical identification of strains of S. aureus, which were then submitted to antibiograms with identification of beta-lactamase producing strains. The results showed contamination in all samples by Staphylococcus spp., with counts ranging from 1.9 x 103 to 5.24 x 106 CFU/mL. Among the antibiotics tested, TOR showed the highest efficiency in elimination or reduction of all S. aureus strains. We found that 50% of S. aureus strains isolated produced the enzyme beta-lactamase. It is necessary to create measures to combat the emergence of new bacterial strains and to promote action by public authorities, supervising the commercialization of veterinary drugs.

Susceptibilidade de Staphylococcus aureus isolados de leite cru a antibióticos comerciais / Susceptibility of Staphylococcus aureus isolated from raw milk to commercial antibiotics

O leite possui extremo valor na dieta humana e constitui um excelente substrato para o crescimento de grande diversidade de microrganismos, dentre eles o Staphylococcus aureus, frequentemente encontrado no leite cru, e que se destaca como um dos microrganismos mais prevalentes em casos de mastite bovina no mundo. Objetivou-se avaliar a sensibilidade de cepas de S. aureus isoladas de leite cru à antimicrobianos comerciais. Para isso, selecionaram-se antibióticos com base em pesquisas junto a estabelecimentos comerciais de medicamentos veterinários no município de Zé Doca, Estado do Maranhão, Brasil. Esses antibióticos foram identificados por associação à base de penicilina, estreptomicina, isoniazida e prednisolona (BPC), oxitetraciclina base (TOR), associação à base de penicilina e estreptomicina (MPP) e cloridrato de tetraciclina (TCA). Posteriormente, realizou-se contagem de Staphylococcus spp., com isolamento e identificação bioquímica das cepas de S. aureus as quais, em seguida, foram submetidas à antibiogramas com identificação de ce,pas produtoras de beta-lactamase. Os resultados demonstraram contaminação em todas as amostras por Staphylococcus spp., com contagens de 1,9 x 103 a 5,24 x 106 UFC/mL. Dentre os antibióticos testados, o TOR apresentou maior eficiência na eliminação ou redução de todas as cepas de S. aureus. Verificou-se que 50% das cepas de S. aureus isoladas produziram enzima beta-lactamase. Torna-se necessário a criação de medidas de combate ao surgimento de novas cepas bacterianas e atuação das autoridades públicas, fiscalizando a comercialização de medicamentos veterinários.(AU)

Milk is extremelly valuable to the human diet and an excellent substrate for the growth of a wide range of microorganisms, including Staphylococcus aureus, frequently found in raw milk and one of the most prevalent microorganisms in cases of bovine mastitis in the world. The objective of this study was to evaluate the sensitivity of S. aureus strains isolated from raw milk to commercial antimicrobials. For this end, antibiotics based on research were selected from commercial establishments of veterinary drugs in the municipality of Zé Doca (Maranhão State, Brazil). The antibiotics were identified by association with penicillin, streptomycin, isoniazid and prednisolone (PCB), oxytetracycline base (TOR), a combination of penicillin and streptomycin (MPP), and tetracycline hydrochloride (TCA). Subsequently, Staphylococcus spp. counting was perfomed, with isolation and biochemical identification of strains of S. aureus, which were then submitted to antibiograms with identification of beta-lactamase producing strains. The results showed contamination in all samples by Staphylococcus spp., with counts ranging from 1.9 x 103 to 5.24 x 106 CFU/mL. Among the antibiotics tested, TOR showed the highest efficiency in elimination or reduction of all S. aureus strains. We found that 50% of S. aureus strains isolated produced the enzyme beta-lactamase. It is necessary to create measures to combat the emergence of new bacterial strains and to promote action by public authorities, supervising the commercialization of veterinary drugs.(AU)

Antimicrobial resistance in ESBL-producing Escherichia coli Isolated from layer and pig farms in Thailand

Background: Study of drug resistance of commensal bacteria in both humans and animals can determine the scale of the drug resistance problem. Usage of antimicrobials to treat infections in humans and animals has generated extensive antimicrobial pressure not only on targeted pathogens but also on commensal bacteria. Commensal Escherichia coli appears to be the major reservoir for resistant genes implicated in the transmission of genetic traits from one bacterium to another. Antimicrobial resistance in Enterobacteriaceae has increased dramatically worldwide in the last decade. An increasing number of community-onset extended-spectrum beta-lactamase (ESBL)-producing bacterial infections, especially those caused by ESBL-producing E. coli, have been reported in many countries, including Thailand. Moreover, ESBL-producing E. coli have been widely detected in food-producing animals and the environment. The increased use of ESBLs in food animals is a serious public health problem. The objective of the study was to determine the prevalence and antimicrobial resistance pattern of ESBL-producing E. coli isolated from pigs, layers, farm workers and stagnant water, in order to increase awareness about antimicrobial usage on farms and to minimize the expansion of the antimicrobial resistance phenomenon in farm settings.Materials, Methods & Results: A total of 588 samples were collected from 107 pig farms and 89 layer farms in Chiang Mai–Lamphun and Chon Buri provinces during May 2015-April 2016. Double-disk diffusion method according to EUCAST (European Committee on Antimicrobial Susceptibility Testing) guidelines was used for detection. The results demonstrated that 36.7% (216/588) of samples were ESBL-producing E. coli-positive, including rectal swabs 74.8% (80/107), pig farm worker stool swabs 57.0% (61/107), stagnant water on pig farms 21.5% (23/107), healthy layer rectal swabs 6.7% (6/89) and layer farm worker stool swabs 51.7% (46/89).[...]

Antimicrobial resistance in ESBL-producing Escherichia coli Isolated from layer and pig farms in Thailand

Background: Study of drug resistance of commensal bacteria in both humans and animals can determine the scale of the drug resistance problem. Usage of antimicrobials to treat infections in humans and animals has generated extensive antimicrobial pressure not only on targeted pathogens but also on commensal bacteria. Commensal Escherichia coli appears to be the major reservoir for resistant genes implicated in the transmission of genetic traits from one bacterium to another. Antimicrobial resistance in Enterobacteriaceae has increased dramatically worldwide in the last decade. An increasing number of community-onset extended-spectrum beta-lactamase (ESBL)-producing bacterial infections, especially those caused by ESBL-producing E. coli, have been reported in many countries, including Thailand. Moreover, ESBL-producing E. coli have been widely detected in food-producing animals and the environment. The increased use of ESBLs in food animals is a serious public health problem. The objective of the study was to determine the prevalence and antimicrobial resistance pattern of ESBL-producing E. coli isolated from pigs, layers, farm workers and stagnant water, in order to increase awareness about antimicrobial usage on farms and to minimize the expansion of the antimicrobial resistance phenomenon in farm settings.Materials, Methods & Results: A total of 588 samples were collected from 107 pig farms and 89 layer farms in Chiang Mai–Lamphun and Chon Buri provinces during May 2015-April 2016. Double-disk diffusion method according to EUCAST (European Committee on Antimicrobial Susceptibility Testing) guidelines was used for detection. The results demonstrated that 36.7% (216/588) of samples were ESBL-producing E. coli-positive, including rectal swabs 74.8% (80/107), pig farm worker stool swabs 57.0% (61/107), stagnant water on pig farms 21.5% (23/107), healthy layer rectal swabs 6.7% (6/89) and layer farm worker stool swabs 51.7% (46/89).[...](AU)

Serogroup identification, phenotypic detection of hemolysis and extended spectrum beta-lactamases of Escherichia coli isolated from psittacine of illegal wildlife trade in Fortaleza, Brazil / Identificação de sorogrupo e detecção fenotípica de betalactamase de espectro estendido e produção de hemolisina em Escherichia coli isoladas de psitacídeos do tráfico de animais silvestres em Fortaleza-CE

This study aimed to identify serogroups of Escherichia coli important for human health in isolates from psittacine of illegal wildlife trade in Ceará State. In addition, hemolysis and production of Extended Spectrum Beta-Lactamases (ESBL) was assessed in the isolates. A total of 78 E. coli strains isolated from different Psittaciformes species from a wildlife rehabilitation center in Fortaleza, Brazil. The isolates used in this study were previously identified and stored. Serogroup identification was performed using polyvalent sera for EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) and EHEC (O157). ESBL detection was performed with double disk synergy method. For hemolysis detection, isolates were inoculated in blood agar base enriched with ovine blood. Only 31 (39.7%) isolates were seropositive and the most frequent were O127, O114, O128 and O111. There was no agglutination for serogroups O55, O124, O136 or O157. Considering both seropositive and seronegative isolates, 9 (11.5%) and 35 (44.9%) presented hemolysis and ESBL production, respectively. In conclusion, the investigated psittacine from illegal wildlife trade hosted ESBL-producing E. coli strains and some belong to important serogroups often linked to severe human infections.(AU)

Este trabalho teve como objetivo identificar sorogrupos de E. coli importantes para a saúde humana, oriundos de psitacídeos provenientes do tráfico no estado do Ceará, assim como detectar atividade hemolítica e produção de betalactamase de espectro estendido (ESBL). Foram testadas 78 cepas de Escherichia coli provenientes de psitaciformes do Centro de Triagem de Animais Silvestres, Fortaleza, CE. Para a identificação dos sorogrupos, utilizaram-se soros polivalentes EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) e EHEC (O157). Para detecção de ESBL, as cepas foram submetidas ao método de aproximação de disco e, para a detecção de hemolisina, foram plaqueadas em ágar sangue base enriquecido com sangue de carneiro. No geral, 31 (39,7%) das amostras foram soropositivas. Os sorogrupos mais frequentemente detectados foram O127, O114, O128 e O111. Não houve positividade para os sorogrupos O55, O124, O136 e O157. Considerando-se as amostras sororreagentes e não sororreagentes, observou-se que nove (11,5%) e 35 (44,9%) cepas de E. coli apresentaram produção de hemolisinas e de ESBL, respectivamente. Em conclusão, constatou-se que psitacídeos provenientes do tráfico de animais silvestres albergam cepas de E. coli produtoras de ESBL e providas de importantes sorogrupos implicados em graves infecções humanas.(AU)

Serogroup identification, phenotypic detection of hemolysis and extended spectrum beta-lactamases of Escherichia coli isolated from psittacine of illegal wildlife trade in Fortaleza, Brazil / Identificação de sorogrupo e detecção fenotípica de betalactamase de espectro estendido e produção de hemolisina em Escherichia coli isoladas de psitacídeos do tráfico de animais silvestres em Fortaleza-CE

This study aimed to identify serogroups of Escherichia coli important for human health in isolates from psittacine of illegal wildlife trade in Ceará State. In addition, hemolysis and production of Extended Spectrum Beta-Lactamases (ESBL) was assessed in the isolates. A total of 78 E. coli strains isolated from different Psittaciformes species from a wildlife rehabilitation center in Fortaleza, Brazil. The isolates used in this study were previously identified and stored. Serogroup identification was performed using polyvalent sera for EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) and EHEC (O157). ESBL detection was performed with double disk synergy method. For hemolysis detection, isolates were inoculated in blood agar base enriched with ovine blood. Only 31 (39.7%) isolates were seropositive and the most frequent were O127, O114, O128 and O111. There was no agglutination for serogroups O55, O124, O136 or O157. Considering both seropositive and seronegative isolates, 9 (11.5%) and 35 (44.9%) presented hemolysis and ESBL production, respectively. In conclusion, the investigated psittacine from illegal wildlife trade hosted ESBL-producing E. coli strains and some belong to important serogroups often linked to severe human infections.(AU)

Este trabalho teve como objetivo identificar sorogrupos de E. coli importantes para a saúde humana, oriundos de psitacídeos provenientes do tráfico no estado do Ceará, assim como detectar atividade hemolítica e produção de betalactamase de espectro estendido (ESBL). Foram testadas 78 cepas de Escherichia coli provenientes de psitaciformes do Centro de Triagem de Animais Silvestres, Fortaleza, CE. Para a identificação dos sorogrupos, utilizaram-se soros polivalentes EPEC (O55, O111, O119, O114, O125, O86, O126, O127, O128), EIEC (O136, O124) e EHEC (O157). Para detecção de ESBL, as cepas foram submetidas ao método de aproximação de disco e, para a detecção de hemolisina, foram plaqueadas em ágar sangue base enriquecido com sangue de carneiro. No geral, 31 (39,7%) das amostras foram soropositivas. Os sorogrupos mais frequentemente detectados foram O127, O114, O128 e O111. Não houve positividade para os sorogrupos O55, O124, O136 e O157. Considerando-se as amostras sororreagentes e não sororreagentes, observou-se que nove (11,5%) e 35 (44,9%) cepas de E. coli apresentaram produção de hemolisinas e de ESBL, respectivamente. Em conclusão, constatou-se que psitacídeos provenientes do tráfico de animais silvestres albergam cepas de E. coli produtoras de ESBL e providas de importantes sorogrupos implicados em graves infecções humanas.(AU)

Draft genome sequence of Chryseobacterium limigenitum SUR2T (LMG 28734T) isolated from dehydrated sludge

The type strain SUR2 of the novel species Chryseobacterium limigenitum was isolated from a dehydrated sludge of the municipal sewage treatment plant in Dogoše near Maribor in Slovenia. The draft genome, with 60 contigs, 4,697,725 bp, 34.4% of G+C content, was obtained using the Illumina HiSeq 2500-1 platform. Joint Genome Institute Microbial Genome Annotation Pipeline (MGAP v.4) has identified 4322 protein-coding sequences including resistance genes against arsenic and other heavy metals. In addition, a subclass B3 metallo-β-lactamase, which confers resistance to penicillins, cephalosporins and carbapenems, was also present in the genome. The genome sequence provides important information regarding bioremediation potential and pathogenic properties of this newly identified species.(AU)