High Genetic Similarity Among Salmonella Heidelberg Isolated from Poultry Farms, Wild Animals, Beef, Poultry and Pork Meat, and Humans in Brazil

Salmonellosis is an important gastrointestinal infection in humans and cause of foodborne outbreaks in the world. In this context, molecular characterization is essential to understand how the strains circulate. The aim of this study was to evaluate the genotypic distribution of S. Heidelberg according to the source of isolation. The genetic relatedness of the S. Heidelberg isolates was determined by pulsed-field gel electrophoresis (PFGE). The most prevalent pulsotypes of cluster A were BRJF6X01.006 (27/95 = 28,42%) related between 1995 and 2011 in broilers, poultry meat and poultry farms, meat product and human, and BRJF6X01.001 (21/95 = 22,10%) related between 2011 and 2017 in wild animals, broilers, poultry meat, poultry farms, meat product, animal feed, and pork meat. The pulsotype BRJF6X01.001 shows a high distribution in the environmental and productive chain. The degree of similarity between pulsotypes BRJF6X01.006 and BRJF6X01.001 is 88%. To ensure the safety of human and animal health, holistic approaches, including surveillance of Salmonella throughout the environment and in the production chain, together with control measures, are critical. As transmission of Salmonella from food producing animals to wildlife and to the environment is considered potential public health problem, information on the survival and persistence of Salmonella in the environment and in potential reservoirs is of considerable importance.(AU)

Genetic Similarities of Escherichia Coli Isolated from Different Substrates of the Broiler Production Chain

Brazil is the largest exporter of chicken meat and poultry farming is one of the most important productive segments, despite major losses due to the bacterium Escherichia coli, which is also a zoonotic microorganism. The objetive of this study was to isolate E. coli and to evaluate its transmissibility potential from the field to chicken meat using the Pulsed Field Gel Electrophoresis (PFGE) technique. Environmental samples (poultry litter, soil and water) were collected from broiler farms located in the South of Brazil where the majority of the Brazilian poultry production occurs. In addition, chicken meat (gizzard, heart, drumette and tulip) samples were collected from local supermarkets. As results, 47.36% of the samples were positives for E. coli. Furthermore, 10 pairs of clones of E. coli were found always in the same substrate (two water-water pairs; three soil-soil pairs and five meat-meat pairs) using PFGE. These findings suggest that certain strains of E. coli may have habitat preferences, making the transfer from one substrate type to another more difficult to occur. Moreover, since no clones were found between environmental samples and chicken meat, it is possible to imply a low risk of E. coli transmissibility throughout the chicken meat production chain.(AU)

Genetic Similarities of Escherichia Coli Isolated from Different Substrates of the Broiler Production Chain

Brazil is the largest exporter of chicken meat and poultry farming is one of the most important productive segments, despite major losses due to the bacterium Escherichia coli, which is also a zoonotic microorganism. The objetive of this study was to isolate E. coli and to evaluate its transmissibility potential from the field to chicken meat using the Pulsed Field Gel Electrophoresis (PFGE) technique. Environmental samples (poultry litter, soil and water) were collected from broiler farms located in the South of Brazil where the majority of the Brazilian poultry production occurs. In addition, chicken meat (gizzard, heart, drumette and tulip) samples were collected from local supermarkets. As results, 47.36% of the samples were positives for E. coli. Furthermore, 10 pairs of clones of E. coli were found always in the same substrate (two water-water pairs; three soil-soil pairs and five meat-meat pairs) using PFGE. These findings suggest that certain strains of E. coli may have habitat preferences, making the transfer from one substrate type to another more difficult to occur. Moreover, since no clones were found between environmental samples and chicken meat, it is possible to imply a low risk of E. coli transmissibility throughout the chicken meat production chain.

Prevalence and serotype distribution of Listeria monocytogenes isolated from foods in Montevideo-Uruguay

ABSTRACT The aim of this work was to study the prevalence of Listeria monocytogenes in foods obtained in retail shops and food industries located in Montevideo-Uruguay, and to identify the serogroups of the obtained isolates. Three-thousand one-hundred and seventy-five food samples (frozen, deli meats, ready-to-eat and cheese) were analyzed. The obtained isolates were serogrouped by multiplex PCR and serotyped by conventional procedure. Genetic comparisons were performed using pulsed-field gel electrophoresis on a sub-set of isolates belonging to the same serotype successively recovered from the same establishment. L. monocytogenes was isolated from 11.2% of samples. The highest prevalence was observed in frozen foods (38%), followed by cheese (10%). 1/2b and 4b were the most frequently identified serotypes. In six of 236 analyzed establishments we successively recovered L. monocytogenes isolates belonging to the same serotype. Most of them corresponded to serotype 1/2b. Pulsed-field gel electrophoresis profiles suggest that at least 33% of L. monocytogenes 1/2b isolates are genetically related and that may remain viable for prolonged periods. The observed prevalence of L. monocytogenes was lower than reported in neighboring countries. Our findings highlight the role that frozen foods may play in the spread of this pathogen, and the relevance of serotypes 1/2b and 4b.(AU)

Prevalence and serotype distribution of Listeria monocytogenes isolated from foods in Montevideo-Uruguay

ABSTRACT The aim of this work was to study the prevalence of Listeria monocytogenes in foods obtained in retail shops and food industries located in Montevideo-Uruguay, and to identify the serogroups of the obtained isolates. Three-thousand one-hundred and seventy-five food samples (frozen, deli meats, ready-to-eat and cheese) were analyzed. The obtained isolates were serogrouped by multiplex PCR and serotyped by conventional procedure. Genetic comparisons were performed using pulsed-field gel electrophoresis on a sub-set of isolates belonging to the same serotype successively recovered from the same establishment. L. monocytogenes was isolated from 11.2% of samples. The highest prevalence was observed in frozen foods (38%), followed by cheese (10%). 1/2b and 4b were the most frequently identified serotypes. In six of 236 analyzed establishments we successively recovered L. monocytogenes isolates belonging to the same serotype. Most of them corresponded to serotype 1/2b. Pulsed-field gel electrophoresis profiles suggest that at least 33% of L. monocytogenes 1/2b isolates are genetically related and that may remain viable for prolonged periods. The observed prevalence of L. monocytogenes was lower than reported in neighboring countries. Our findings highlight the role that frozen foods may play in the spread of this pathogen, and the relevance of serotypes 1/2b and 4b.

Pertussis characterization in the central-west region of São Paulo state, Brazil / Caracterização da coqueluche na região centro-oeste do Estado de São Paulo, Brasil

Pertussis is a highly contagious respiratory disease caused by Bordetella pertussis. This study aimed at characterizing the B. pertussis laboratory positivity and the isolated strains in municipalities of the Central-West Region of São Paulo State, Brazil from 2010 to 2014. A total of 597 nasopharyngeal swabs samples were collected from suspected patients and contacts, and analyzed by in vitro culture and Real-Time PCR (qPCR). Culture-positive B. pertussis strains were characterized by serotyping and pulsed-field gel electrophoresis. Considering culture and/or qPCR, the positivity rate was of 19.6%. Out of 117 samples with B. pertussis, 23 were detected by both methods, 89 by qPCR only and five by culture only. Strains presenting FIM3 (40%), FIM2,3 (32%) and FIM2 (28%) serotypes were found. Five pulsotypes were detected by PFGE, 48% of which identified as BP.Xba.0039, being the predominant type in this study. Among the positive strains, 50% were isolated from <2 months old-children and 17% were isolated from three to six months old patients. Non-vaccinated children or with incomplete vaccination schedule were at the major risk of complications and death, highlighting the importance of a continuous monitoring of this infection for the future control strategies.

A coqueluche é uma doença respiratória altamente contagiosa causada por Bordetella pertussis. Este estudo caracterizou a positividade de B. pertussis e as cepas isoladas em municípios da Região Centro-Oeste do Estado de São Paulo de 2010 a 2014. Foram coletados 597 esfregaços nasofaríngeos de pacientes e contatos suspeitos de coqueluche, e analisados por cultura e Real-TimePCR (qPCR). Os isolados de B. pertussis obtidos de cultura foram caracterizados por sorotipagem e eletroforese em gel de campo pulsado. Considerando-se a cultura e/ou qPCR, verificou-se taxa de positividade de 19,6%. Das 117 amostras positivas para B. pertussis, 23 foram detectadas por ambos os métodos, 89 apenas por qPCR e cinco apenas na cultura. Foram detectadas cepas de sorotipos FIM3 (40%), FIM2,3 (32%) e FIM2 (28%). Cinco pulsotipos foram detectados pela PFGE, e 48% identificados como BP.Xba.0039, o tipo predominante neste estudo. Entre as cepas positivas, 50% foram isoladas de crianças menores de dois meses e 17% isoladas da faixa etária de três a seis meses. Crianças não vacinadas ou com esquema de vacinação incompleta têm maior risco de complicações e óbito, o que ressalta a importância do monitoramento contínuo desta infecção para futuras estratégias de controle.

Pertussis characterization in the central-west region of São Paulo state, Brazil / Caracterização da coqueluche na região centro-oeste do Estado de São Paulo, Brasil

Pertussis is a highly contagious respiratory disease caused by Bordetella pertussis. This study aimed at characterizing the B. pertussis laboratory positivity and the isolated strains in municipalities of the Central-West Region of São Paulo State, Brazil from 2010 to 2014. A total of 597 nasopharyngeal swabs samples were collected from suspected patients and contacts, and analyzed by in vitro culture and Real-Time PCR (qPCR). Culture-positive B. pertussis strains were characterized by serotyping and pulsed-field gel electrophoresis. Considering culture and/or qPCR, the positivity rate was of 19.6%. Out of 117 samples with B. pertussis, 23 were detected by both methods, 89 by qPCR only and five by culture only. Strains presenting FIM3 (40%), FIM2,3 (32%) and FIM2 (28%) serotypes were found. Five pulsotypes were detected by PFGE, 48% of which identified as BP.Xba.0039, being the predominant type in this study. Among the positive strains, 50% were isolated from <2 months old-children and 17% were isolated from three to six months old patients. Non-vaccinated children or with incomplete vaccination schedule were at the major risk of complications and death, highlighting the importance of a continuous monitoring of this infection for the future control strategies.(AU)

A coqueluche é uma doença respiratória altamente contagiosa causada por Bordetella pertussis. Este estudo caracterizou a positividade de B. pertussis e as cepas isoladas em municípios da Região Centro-Oeste do Estado de São Paulo de 2010 a 2014. Foram coletados 597 esfregaços nasofaríngeos de pacientes e contatos suspeitos de coqueluche, e analisados por cultura e Real-TimePCR (qPCR). Os isolados de B. pertussis obtidos de cultura foram caracterizados por sorotipagem e eletroforese em gel de campo pulsado. Considerando-se a cultura e/ou qPCR, verificou-se taxa de positividade de 19,6%. Das 117 amostras positivas para B. pertussis, 23 foram detectadas por ambos os métodos, 89 apenas por qPCR e cinco apenas na cultura. Foram detectadas cepas de sorotipos FIM3 (40%), FIM2,3 (32%) e FIM2 (28%). Cinco pulsotipos foram detectados pela PFGE, e 48% identificados como BP.Xba.0039, o tipo predominante neste estudo. Entre as cepas positivas, 50% foram isoladas de crianças menores de dois meses e 17% isoladas da faixa etária de três a seis meses. Crianças não vacinadas ou com esquema de vacinação incompleta têm maior risco de complicações e óbito, o que ressalta a importância do monitoramento contínuo desta infecção para futuras estratégias de controle.(AU)

Caracterização molecular de Listeria monocytogenes oriundas de cortes cárneos bovinos e de abatedouros frigoríficos de bovinos localizados no Distrito Federal, Brasil / Molecular characterization of Listeria monocytogenes from beef samples and cattle slaughterhouses located in the Federal District, Brazil

Este trabalho teve como objetivo realizar a detecção de cepas de Listeria monocytogenes de cortes cárneos bovinos bem como no ambiente de abatedouros frigoríficos localizados no Distrito Federal, promover a sorotipificação pela reação em cadeia da polimerase (PCR), realizar antibiograma e submeter às cepas à eletroforese de campo pulsado (Pulsed-field gel electrophoresis - PFGE). Foram analisados um total de 125 cortes cárneos bovinos, 45 amostras de swabs de carcaças e 43 amostras de swabs em que foram detectados 13 cepas de Listeria monocytogenes, sendo 11 em cortes cárneos bovinos e 2 swabs de ambiente em um abatedouro frigorifico. Não foram isoladas cepas de swabs de carcaça. Dentre as 13 cepas de Listeria monocytogenes foram encontradas seis cepas do sorotipo 4b, cinco do sorotipo 1/2c e duas cepas do sorotipo 1/2a. Dentre as 11 cepas de L. monocytogenes encontradas em cortes cárneos bovino, uma (9,1%) cepa apresentou resistência a eritromicina, outra (9,1%) cepa a gentamicina e outra a ciprofloxacina (9,1%) e todas as cepas (100%) apresentaram resistência ao Ác. Nalidíxico. Das duas (2) cepas oriundas de ralos de abatedouro frigorífico, todas (100%) apresentaram resistência ao Ác. Nalidíxico e a sulfonamidas. A análise por eletroforese de campo pulsante (PFGE) demonstrou 13 diferentes pulsotipos, em que foram agrupados em 3 diferentes grupos clonais, que coincidentemente se correlacionavam com os 3 diferentes sorotipos encontrados sugerindo uma ampla disseminação desses perfis no Distrito Federal.(AU)

The aim of the study was the analysis of Listeria monocytogenes strains in beef samples as well as slaughterhouse environment, located in the Federal District, promote serotyping by polymerase chain reaction (PCR), perform antibiotic susceptibility and submit the strains to Pulsed-field gel electrophoresis (PFGE). A total of 125 beef samples were analyzed, 45 samples of carcasses swabs and 43 swab samples. It detected 13 strains of Listeria monocytogenes, 11 in beef samples. and 2 in slaughterhouse environment. No carcass swabs strains were isolated. Among the 13 strains of L. monocytogenes six strains of serotype 4b were found, five serotype 1/2c and two strains of serotype 1/2a. Among the 11 strains of L. monocytogenes found in beef, one (9.1%) strain showed resistance to erythromycin, one (9.1%) strain to gentamicin, one to ciprofloxacin (9.1%) and all strains (100%) were resistant to nalidixic acid. The two strains coming from the slaughterhouse drains, all (100%) were resistant to nalidixic acid and Sulfonamides. The analysis by pulsed field gel electrophoresis (PFGE) showed 13 different pulsotypes; they were grouped into three different clonal groups, coincidentally correlated with the three different serotypes found, what suggests a widespread dissemination of these profiles in the Federal District, Brazil.(AU)

Caracterização molecular de Listeria monocytogenes oriundas de cortes cárneos bovinos e de abatedouros frigoríficos de bovinos localizados no Distrito Federal, Brasil / Molecular characterization of Listeria monocytogenes from beef samples and cattle slaughterhouses located in the Federal District, Brazil

The aim of the study was the analysis of Listeria monocytogenes strains in beef samples as well as slaughterhouse environment, located in the Federal District, promote serotyping by polymerase chain reaction (PCR), perform antibiotic susceptibility and submit the strains to Pulsed-field gel electrophoresis (PFGE). A total of 125 beef samples were analyzed, 45 samples of carcasses swabs and 43 swab samples. It detected 13 strains of Listeria monocytogenes, 11 in beef samples. and 2 in slaughterhouse environment. No carcass swabs strains were isolated. Among the 13 strains of L. monocytogenes six strains of serotype 4b were found, five serotype 1/2c and two strains of serotype 1/2a. Among the 11 strains of L. monocytogenes found in beef, one (9.1%) strain showed resistance to erythromycin, one (9.1%) strain to gentamicin, one to ciprofloxacin (9.1%) and all strains (100%) were resistant to nalidixic acid. The two strains coming from the slaughterhouse drains, all (100%) were resistant to nalidixic acid and Sulfonamides. The analysis by pulsed field gel electrophoresis (PFGE) showed 13 different pulsotypes; they were grouped into three different clonal groups, coincidentally correlated with the three different serotypes found, what suggests a widespread dissemination of these profiles in the Federal District, Brazil.(AU)

Este trabalho teve como objetivo realizar a detecção de cepas de Listeria monocytogenes de cortes cárneos bovinos bem como no ambiente de abatedouros frigoríficos localizados no Distrito Federal, promover a sorotipificação pela reação em cadeia da polimerase (PCR), realizar antibiograma e submeter às cepas à eletroforese de campo pulsado (Pulsed-field gel electrophoresis - PFGE). Foram analisados um total de 125 cortes cárneos bovinos, 45 amostras de swabs de carcaças e 43 amostras de swabs em que foram detectados 13 cepas de Listeria monocytogenes, sendo 11 em cortes cárneos bovinos e 2 swabs de ambiente em um abatedouro frigorifico. Não foram isoladas cepas de swabs de carcaça. Dentre as 13 cepas de Listeria monocytogenes foram encontradas seis cepas do sorotipo 4b, cinco do sorotipo 1/2c e duas cepas do sorotipo 1/2a. Dentre as 11 cepas de L. monocytogenes encontradas em cortes cárneos bovino, uma (9,1%) cepa apresentou resistência a eritromicina, outra (9,1%) cepa a gentamicina e outra a ciprofloxacina (9,1%) e todas as cepas (100%) apresentaram resistência ao Ác. Nalidíxico. Das duas (2) cepas oriundas de ralos de abatedouro frigorífico, todas (100%) apresentaram resistência ao Ác. Nalidíxico e a sulfonamidas. A análise por eletroforese de campo pulsante (PFGE) demonstrou 13 diferentes pulsotipos, em que foram agrupados em 3 diferentes grupos clonais, que coincidentemente se correlacionavam com os 3 diferentes sorotipos encontrados sugerindo uma ampla disseminação desses perfis no Distrito Federal.(AU)

CONTAMINAÇÃO POR ENTEROBACTÉRIAS RESISTENTES A ANTIMICROBIANOS ISOLADAS EM CELULARES E MÃOS EM HOSPITAL VETERINÁRIO

As infecções nosocomiais representam sério problema de saúde pública, levando ao aumento da mortalidade e morbidade dos pacientes afetados. Os telefones celulares são alvo de investigações sobre sua importância na disseminação de patógenos, devido ao seu uso descontrolado e presença em todos os ambientes do dia a dia, incluindo hospitais veterinários. Nesse sentido, o objetivo deste trabalho é avaliar a presença de Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis) e Klebsiella pneumoniae (K. pneumoniae), importantes bactérias associadas à infecção hospitalar, em telefones celulares da equipe cirúrgica de pequenos animais do Hospital Veterinário da Universidade Estadual Paulista Câmpus Jaboticabal. Para isso, foram coletadas amostras de telefones celulares dos cirurgiões, auxiliares, enfermeiros e anestesistas e dos responsáveis pelos pacientes, bem como também de suas mãos, mesa cirúrgica e do paciente. As bactérias foram isoladas e identificadas através do teste de reação em cadeia de polimerase (PCR), a diversidade genética por Eletroforese em gel de Campo Pulsado (PFGE) e a suscetibilidade à antimicrobianos através de antibiograma. Adicionalmente, os pacientes foram acompanhados por 4 semanas quanto à sinais de infecção nosocomial na ferida operatória. Oito amostras foram positivas para E. coli e Proteus mirabilis, provenientes da mão do anestesista, mão do tutor, mão do cirurgião, celular do cirurgião, celular do enfermeiro, celular do anestesista e duas mesas cirúrgicas. O teste de sensibilidade antimicrobiana demonstrou que todos os isolados eram resistentes à múltiplos antimicrobianos. A eletroforese de campo pulsado demonstrou alta diversidade genética entre os isolados. A identificação de E.coli e Proteus mirabilis multirresistentes em aparelhos celulares da equipe cirúrgica representa grande preocupação e, apesar de não diretamente correlacionado, o isolamento dessas bactérias no interior da área limpa do centro cirúrgico evidenciou a possibilidade de transmissão nosocomial a partir dos aparelhos celulares à pacientes susceptíveis.

Nosocomial infections represent a serious public health problem, leading to increased mortality and morbidity for affected patients. Cell phones are the target of investigations about their importance in the spread of pathogens, due to their uncontrolled use and presence in all day-to-day environments, including veterinary hospitals. In this sense, the objective of this work is to evaluate the presence of Escherichia coli (E.coli), Proteus mirabilis (P. mirabilis) and Klebsiella pneumoniae (K. pneumoniae), important bacteria associated with hospital infection, on cell phones of the small animal surgical team at the Veterinary Hospital of Paulista State University - Campus Jaboticabal. For this, samples of cell phones were collected from surgeons, assistants, nurses and anesthetists and those responsible for patients, as well as from their hands, operating table and the patient. The bacteria were isolated and identified through the polymerase chain reaction (PCR) test, genetic diversity by Pulsed Field Gel Electrophoresis (PFGE) and susceptibility to antimicrobials through antibiogram. Additionally, the patients were followed up for 4 weeks for signs of nosocomial infection in the surgical wound. Eight samples were positive for E. coli and Proteus mirabilis, from the hand of the anesthetist, hand of the tutor, hand of the surgeon, cell of the surgeon, cell of the nurse, cell of the anesthetist and two surgical tables. The antimicrobial susceptibility test demonstrated that all isolates were resistant to multiple antimicrobials. Pulsed field electrophoresis demonstrated high genetic diversity among the isolates. The identification of multi- resistant E.coli and Proteus mirabilis in cell phones of the surgical team is of great concern and, although not directly correlated, the isolation of these bacteria within the clean area of the operating room has evidenced the possibility of nosocomial transmission from cell phones susceptible patients.

Drug resistance profiles and clonality of sporadic Shigella sonnei isolates in Ankara, Turkey

The aims of this study were to investigate drug resistance rates, types of extended spectrum beta lactamases (ESBLs), and molecular epidemiological characteristics of 43 Shigella sonnei isolates. Ampicillin-sulbactam, amoxicillin-clavulanate, chloramphenicol, and ciprofloxacin were the most active antibiotics. Five isolates harbored blaSHV-12, blaTEM-1 and blaCTX-M-15. More than 90% of the isolates had an indistinguishable pulsotype.

Genetic profiling of Klebsiella pneumoniae: comparison of pulsed field gel electrophoresis and random amplified polymorphic DNA

In this study, the discriminatory power of pulsed field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD) methods for subtyping of 54 clinical isolates of Klebsiella pneumoniae were compared. All isolates were typeable by RAPD, while 3.6% of them were not typeable by PFGE. The repeatability of both typing methods were 100% with satisfying reproducibility ( 95%). Although the discriminatory power of PFGE was greater than RAPD, both methods showed sufficient discriminatory power (DI > 0.95) which reflects the heterogeneity among the K. pneumoniae isolates. An optimized RAPD protocol is less technically demanding and time consuming that makes it a reliable typing method and competitive with PFGE.(AU)

Molecular characterization of Listeria monocytogenes isolated from animal products in a city of Northern Brazil

Listeria monocytogenes, a foodborne pathogen causes listeriosis, a fatal disease in about 30% of cases that affects mainly immunocompromised persons. The aim of this research was to characterize L. monocytogenes pulsed-field gel electrophoresis (PFGE) types isolated from meat products collected at public markets in Araguaina city, TO. Sixty samples of raw ground beef and frescal sausage were analyzed during the second half of 2008. Five out of 30 samples (16.7%) of raw ground beef tested positive for L. monocytogenes, three of which were classified as serotype 1/2b and two as serotype 4b. Among the 30 samples of sausage collected, two strains of L. monocytogenes were isolated (6.7%), one of them belonging to serotype 1/2a and the other belonging to serotype 1/2b. The restriction enzymes used were ApaI and SmaI. Similarities among the strains were determined by Dice coefficient. The macro restriction profile obtained by using SmaI enzyme allowed the distribution of seven strains in two clusters, two pulsotypes and two subtypes. The result indicates that L. monocytogenes isolates, belonging to serotype 4b, 1/2a and 1/2b, are strongly correlated within the same serotype group, and in some cases among different serotypes, suggesting that they have a common source.

Listeria monocytogenes é um patógeno de origem alimentar que causa a listeriose, doença fatal em aproximadamente 30% dos casos, e que afeta principalmente pessoas imunocomprometidas. O presente trabalho teve como objetivo analisar os perfis PFGE de cepas de L. monocytogenes isoladas de produtos de origem animal, obtidos em mercados públicos da cidade de Araguaína, TO. Foram analisadas 60 amostras de carne moída crua e de linguiça frescal, no segundo semestre de 2008. Cinco (16,7%) das 30 amostras de carne moída crua foram positivas ao patógeno, sendo que três pertenciam ao sorotipo 1/2b e duas ao sorotipo 4b. Das 30 amostras de linguiça mista frescal, duas (6,7%) foram positivas para L. monocytogenes, sendo uma do sorotipo 1/2a e outra do 1/2b. Foram utilizadas as enzimas de restrição ApaI e SmaI. A similaridade entre eles foi determinada pelo coeficiente de Dice. A análise do perfil de macrorestrição com a enzima SmaI permitiu a distribuição dos sete isolados em dois clusters, dois pulsotipos e dois subtipos. Os resultados permitiram concluir que os isolados de L. monocytogenes sorotipos 4b, 1/2a e 1/2b foram fortemente correlacionados dentro dos mesmos sorotipos e em alguns casos entre diferentes sorotipos, sugerindo uma fonte comum.

Molecular characterization of Listeria monocytogenes isolated from animal products in a city of Northern Brazil

Listeria monocytogenes, a foodborne pathogen causes listeriosis, a fatal disease in about 30% of cases that affects mainly immunocompromised persons. The aim of this research was to characterize L. monocytogenes pulsed-field gel electrophoresis (PFGE) types isolated from meat products collected at public markets in Araguaina city, TO. Sixty samples of raw ground beef and frescal sausage were analyzed during the second half of 2008. Five out of 30 samples (16.7%) of raw ground beef tested positive for L. monocytogenes, three of which were classified as serotype 1/2b and two as serotype 4b. Among the 30 samples of sausage collected, two strains of L. monocytogenes were isolated (6.7%), one of them belonging to serotype 1/2a and the other belonging to serotype 1/2b. The restriction enzymes used were ApaI and SmaI. Similarities among the strains were determined by Dice coefficient. The macro restriction profile obtained by using SmaI enzyme allowed the distribution of seven strains in two clusters, two pulsotypes and two subtypes. The result indicates that L. monocytogenes isolates, belonging to serotype 4b, 1/2a and 1/2b, are strongly correlated within the same serotype group, and in some cases among different serotypes, suggesting that they have a common source.

Listeria monocytogenes é um patógeno de origem alimentar que causa a listeriose, doença fatal em aproximadamente 30% dos casos, e que afeta principalmente pessoas imunocomprometidas. O presente trabalho teve como objetivo analisar os perfis PFGE de cepas de L. monocytogenes isoladas de produtos de origem animal, obtidos em mercados públicos da cidade de Araguaína, TO. Foram analisadas 60 amostras de carne moída crua e de linguiça frescal, no segundo semestre de 2008. Cinco (16,7%) das 30 amostras de carne moída crua foram positivas ao patógeno, sendo que três pertenciam ao sorotipo 1/2b e duas ao sorotipo 4b. Das 30 amostras de linguiça mista frescal, duas (6,7%) foram positivas para L. monocytogenes, sendo uma do sorotipo 1/2a e outra do 1/2b. Foram utilizadas as enzimas de restrição ApaI e SmaI. A similaridade entre eles foi determinada pelo coeficiente de Dice. A análise do perfil de macrorestrição com a enzima SmaI permitiu a distribuição dos sete isolados em dois clusters, dois pulsotipos e dois subtipos. Os resultados permitiram concluir que os isolados de L. monocytogenes sorotipos 4b, 1/2a e 1/2b foram fortemente correlacionados dentro dos mesmos sorotipos e em alguns casos entre diferentes sorotipos, sugerindo uma fonte comum.

Molecular epidemiology and extended-spectrum Beta-lactamases production of Klebsiella pneumoniae isolated from three dairy herds / Epidemiologia molecular e produção de Beta- lac-tamases de espectro estendido de Klebsiella pneumoniae isoladas de três propriedades leiteiras

The objectives of this study were to isolate Klebsiella pneumoniae from different sources in three dairy cattle herds, to use the pulsed-field gel electrophoresis (PFGE) to measure genotypic similarities between isolates within a dairy herd, to verify the production of extended-spectrum β-lactamases (ESBLs) by the double-disk synergy test (DDST), and to use the PCR to detect the main ESBLs subgroups genes. Three dairy farms were selected based on previous mastitis outbreaks caused by K. pneumoniae. Milk samples were collected from lactating cows and from the bulk tank. Swabs were performed in different locations, including milking parlors, waiting room, soil, animal's hind limbs and rectum. K. pneumoniae was isolated from 27 cases of intramammary infections (IMI) and from 41 swabs. For farm A isolates from IMI and bulk tank were considered of the same PGFE subtype. One isolate from a bulk tank, three from IMI cases and four from environmental samples were positive in the DDST test. All eight DDST positive isolates harbored the bla shv gene, one harbored the bla tem gene, and three harbored the bla ctx-m gene, including the bulk tank isolate. Our study confirms that ESBL producing bacteria is present in different locations in dairy farms, and may be responsible for IMI. The detection of ESBLs on dairy herds could be a major concern for both public and animal health.(AU)

Os objetivos deste estudo foram isolar Klebsiella pneumoniae de diferentes localidades em três propriedades leiteiras, utilizar a eletroforese em campo pulsátil para averiguar similaridades genotípicas entre os isolados de uma mesma propriedade, verificar a produção de beta-lactamases de espectro estendido (ESBLs) pela prova da disco-difusão dupla associada (DDST) e utilizar a PCR para detecção dos principais subgrupos genéticos de ESBLs. Três propriedades leiteiras foram selecionadas baseando-se em surtos prévios de mastites causadas por K. pneumoniae. Amostras de leite foram coletadas de vacas em lactação e do tanque de expansão. Swabs foram realizados em diferentes localidades, incluindo salas de lactação, salas de espera, solo, reto e membros posteriores de animais. K. pneumoniae foi isolada de 27 casos de infecções intramamária (IMI) e de 41 swabs. Para a propriedade A os isolados de IMI e do tanque de expansão foram considerados do mesmo subtipo molecular. Um isolado do tanque de expansão, três de casos de IMI e quatro de amostras ambientais foram considerados positivos no teste da DDST. Todos os oito isolados DDST positivos portavam o gene bla shv, um portava o gene bla tem, e três portavam o gene bla ctx-m, incluindo um isolado de tanque de expansão. Nosso estudo confirma que bactérias produtoras de ESBLs estão presentes em diferentes localidades em propriedades leiteiras, e podem ser responsáveis por quadros de IMI. A detecção de ESBLs em propriedades leiteiras pode representar uma grande preocupação para saúde pública e para a saúde animal.(AU)

Antimicrobial resistance profile, phage typing and molecular characterization of Salmonella Enteritidis strains from poultry origin / Perfil de resistência a antimicrobianos, fagotipagem e caracterização molecular de cepas de Salmonella Enteritidis de origem avícola

Salmonella enterica subsp. enterica serovar Enteritidis (SE) is currently responsible for significant losses in the poultry industry with consequences on public health. In the present study, 38 SE isolates from biological material of chicken breeders were characterized using phage typing, antimicrobial resistance profile and pulsed field gel electrophoresis (PFGE). The phage type (PT) 7 and 9 were predominant and 26.3% of the isolates were resistance to nalidixic acid (NAL). The phage typing and analysis of antimicrobial resistance profile showed high discriminatory power (0.85). The PFGE profile of 13 strains with XbaI and SpeI discriminated the SE phage types, as well characterized strains from the same serovar. The results showed the importance to associate different methods for the characterization of SE and suggest that poultry products may have been source of human salmonellosis in the Parana State during the period of study.

Salmonella enterica subsp. enterica sorovar Enteritidis (SE) é responsável por prejuízos significativos à avicultura, com conseqüências importantes para saúde pública. No presente estudo foi realizada a caracterização de 38 isolados de SE provenientes de material biológico de reprodutoras destinadas à produção de frangos de corte através de fagotipagem, perfil de resistência antimicrobiana e de eletrofose de campo pulsado (PFGE). Os fagotipos (PT) predominantes foram PT7 e PT9 e 26,3% dos isolados apresentaram resistência ao ácido nalidíxico (NAL). A fagotipagem e a análise do perfil de resistência antimicrobiana juntos mostraram elevado poder discriminatório (0,85). Os perfis de PFGE de 13 cepas, com as endonucleases XbaI e SpeI permitiram discriminar os fagotipos de SE, bem como associar perfis de um mesmo fagotipo. Os resultados mostram a importância da associação de métodos para a caracterização de SE e sugerem que produtos da avicultura de corte podem ser fonte de salmonelose humana.

Antimicrobial resistance profile, phage typing and molecular characterization of Salmonella Enteritidis strains from poultry origin / Perfil de resistência a antimicrobianos, fagotipagem e caracterização molecular de cepas de Salmonella Enteritidis de origem avícola

Salmonella enterica subsp. enterica serovar Enteritidis (SE) is currently responsible for significant losses in the poultry industry with consequences on public health. In the present study, 38 SE isolates from biological material of chicken breeders were characterized using phage typing, antimicrobial resistance profile and pulsed field gel electrophoresis (PFGE). The phage type (PT) 7 and 9 were predominant and 26.3% of the isolates were resistance to nalidixic acid (NAL). The phage typing and analysis of antimicrobial resistance profile showed high discriminatory power (0.85). The PFGE profile of 13 strains with XbaI and SpeI discriminated the SE phage types, as well characterized strains from the same serovar. The results showed the importance to associate different methods for the characterization of SE and suggest that poultry products may have been source of human salmonellosis in the Parana State during the period of study. 

Salmonella enterica subsp. enterica sorovar Enteritidis (SE) é responsável por prejuízos significativos à avicultura, com conseqüências importantes para saúde pública. No presente estudo foi realizada a caracterização de 38 isolados de SE provenientes de material biológico de reprodutoras destinadas à produção de frangos de corte através de fagotipagem, perfil de resistência antimicrobiana e de eletrofose de campo pulsado (PFGE). Os fagotipos (PT) predominantes foram PT7 e PT9 e 26,3% dos isolados apresentaram resistência ao ácido nalidíxico (NAL). A fagotipagem e a análise do perfil de resistência antimicrobiana juntos mostraram elevado poder discriminatório (0,85). Os perfis de PFGE de 13 cepas, com as endonucleases XbaI e SpeI permitiram discriminar os fagotipos de SE, bem como associar perfis de um mesmo fagotipo. Os resultados mostram a importância da associação de métodos para a caracterização de SE e sugerem que produtos da avicultura de corte podem ser fonte de salmonelose humana.