Antimicrobial Resistance in Salmonella Serovars Isolated From an Egg-Producing Region in Brazil

Fowl paratyphoid infections are caused by different Salmonella serovars that can affect a wide range of hosts. Due to its complex epidemiology, Salmonella serovar identification is crucial for the development and implementation of monitoring and control programs in poultry farms. Moreover, the characterization of the antimicrobial resistance profiles of Salmonella strains isolated from livestock is relevant to public health because they are a common causative agent of foodborne diseases. The objective of this study was to investigate the presence of Salmonella spp. and to identify the antimicrobial resistance profiles of strains isolated in the midwestern region of São Paulo state, which accounts for the highest production of table eggs in Brazil. For this purpose, 2008 fecal samples were collected on 151 commercial layer farms and submitted to microbiological analyses. Twenty-two serovars were isolated from 80 (52.9%) farms, among which S. Mbandaka and S. Braenderup were the most prevalent. All isolates expressed resistance to at least one of the 23 antimicrobials tested, and the highest resistance rates were determined against streptomycin (93.5%) and sulfonamide (84.6%). Moreover, multidrug resistance was observed in 41% of the isolates and the maximum drug resistance profile was against ten different antimicrobials. Therefore, the identification of Salmonella serovars in poultry production provides epidemiological knowledge to develop prevention and control measures in order to ensure poultry health and to prevent human infection by multiresistant strains.

Antimicrobial Resistance in Salmonella Serovars Isolated From an Egg-Producing Region in Brazil

Fowl paratyphoid infections are caused by different Salmonella serovars that can affect a wide range of hosts. Due to its complex epidemiology, Salmonella serovar identification is crucial for the development and implementation of monitoring and control programs in poultry farms. Moreover, the characterization of the antimicrobial resistance profiles of Salmonella strains isolated from livestock is relevant to public health because they are a common causative agent of foodborne diseases. The objective of this study was to investigate the presence of Salmonella spp. and to identify the antimicrobial resistance profiles of strains isolated in the midwestern region of São Paulo state, which accounts for the highest production of table eggs in Brazil. For this purpose, 2008 fecal samples were collected on 151 commercial layer farms and submitted to microbiological analyses. Twenty-two serovars were isolated from 80 (52.9%) farms, among which S. Mbandaka and S. Braenderup were the most prevalent. All isolates expressed resistance to at least one of the 23 antimicrobials tested, and the highest resistance rates were determined against streptomycin (93.5%) and sulfonamide (84.6%). Moreover, multidrug resistance was observed in 41% of the isolates and the maximum drug resistance profile was against ten different antimicrobials. Therefore, the identification of Salmonella serovars in poultry production provides epidemiological knowledge to develop prevention and control measures in order to ensure poultry health and to prevent human infection by multiresistant strains.(AU)

Highly Pathogenic and Multidrug Resistant Avian Pathogenic Escherichia Coli in Free-Range Chickens from Brazil

Avian pathogenic Escherichiacoli (APEC) virulence mechanism has been continuously studied and it is believed to be multifactorial and because of this, this work aimed to characterize potentially APEC strains isolated from free-range hens. Isolates were submitted to PCR for the detection of virulence genes, which were of high prevalence. In vivo inoculation of day-old chicks revealed that 49 of these strains were of high and intermediate pathogenicity. In addition, isolates were submitted to antimicrobials susceptibility test with the majority of the strains presenting multiresistance. Phylogenetic analysis showed a greater presence of potentially APEC isolates in-group B2. In addition, high heterogeneity was detected among the isolates byXbaI enzyme. Fifteen serogroups were identified, being the O8 the most frequent. These results strengthen the fact that a combination of diverse factors are associated with the pathogenicity APEC strains, as well as to highlight its importance to public health and that free-range hens can act as a reservoirs of potentially zoonoticbacteria.(AU)

Highly Pathogenic and Multidrug Resistant Avian Pathogenic Escherichia Coli in Free-Range Chickens from Brazil

Avian pathogenic Escherichiacoli (APEC) virulence mechanism has been continuously studied and it is believed to be multifactorial and because of this, this work aimed to characterize potentially APEC strains isolated from free-range hens. Isolates were submitted to PCR for the detection of virulence genes, which were of high prevalence. In vivo inoculation of day-old chicks revealed that 49 of these strains were of high and intermediate pathogenicity. In addition, isolates were submitted to antimicrobials susceptibility test with the majority of the strains presenting multiresistance. Phylogenetic analysis showed a greater presence of potentially APEC isolates in-group B2. In addition, high heterogeneity was detected among the isolates byXbaI enzyme. Fifteen serogroups were identified, being the O8 the most frequent. These results strengthen the fact that a combination of diverse factors are associated with the pathogenicity APEC strains, as well as to highlight its importance to public health and that free-range hens can act as a reservoirs of potentially zoonoticbacteria.

Multidrug-resistant pathogenic Escherichia coli isolated from wild birds in a veterinary hospital.

Wild birds are carriers of Escherichia coli. However, little is known about their role as reservoirs for extra-intestinal pathogenic E. coli (ExPEC). In this work we investigated E. coli strains carrying virulence genes related to human and animal ExPEC isolated from free-living wild birds treated in a veterinary hospital. Multidrug resistance was found in 47.4% of the strains, but none of them were extended-spectrum beta-lactamase producers. Not only the virulence genes, but also the serogroups (e.g. O1 and O2) detected in the isolates of E. coli have already been implicated in human and bird diseases. The sequence types detected were also found in wild, companion and food animals, environmental and human clinical isolates in different countries. Furthermore, from the 19 isolates, 17 (89.5%) showed a degree of pathogenicity on an in vivo infection model. The isolates showed high heterogeneity by pulsed-field gel electrophoresis indicating that E. coli from these birds are clonally diverse. Overall, the results showed that wild birds can be reservoirs and/or vectors of highly pathogenic and multidrug-resistant E. coli that have the potential to cause disease in humans and poultry.

Comparison between avian pathogenic (APEC) and avian faecal (AFEC) Escherichia coli isolated from different regions in Brazil.

Detection and analysis of virulence-associated genes (VAGs) of avian pathogenic Escherichia coli (APEC) may be helpful to distinguish pathogenic from commensal faecal strains (AFEC). The aim of this study was to characterise 120 isolates of avian Escherichia coli, comprising 91 APEC (from diseased birds) and 29 AFEC (from healthy chickens), collected in Brazil. Phylogenetic analysis and in vivo pathogenicity testing was performed on 38 VAGs. The VAGs iucD, iutA, iroN, fepC, ompT, cvi and hlyF were statistically associated with medium and high pathogenicity (MP/HP) strains. A minimal group of seven VAGs may be required to accurately discriminate pathogenic and non-pathogenic avian strains of E. coli in Brazil.

Evaluation of Disinfectants Used in Pre-Chilling water Tanks of Poultry Processing Plants

In poultry processing plants, disinfectants are often added to pre-chilling water tanks to reduce microbial contamination. The present study aimed at evaluating the effect of five disinfectants (acidified sodium chlorite, alkyl dimethyl benzyl ammonium chloride, chlorine dioxide, peracetic acid, and sodium hypochlorite) on the populations of food quality indicator microorganisms and on Salmonella Enteritidis (SE) in the presence and absence of organic matter. The results showed that chlorine dioxide and sodium hypochlorite did not reduce microbial carcass counts. On the other hand, acidified sodium chlorite, alkyl dimethyl benzyl ammonium chloride and peracetic acid reduced total and fecal coliform counts. Peracetic acid reduced the number of psychrotrophic microorganisms. All products were effective in reducing SE counts only in the absence of organic matter. Acidified sodium chlorite, alkyl dimethyl benzyl ammonium chloride and peracetic acid could be candidates for the replacement of sodium hypochlorite (commonly used in Brazil) in pre-chilling tanks. (AU)

Evaluation of Disinfectants Used in Pre-Chilling water Tanks of Poultry Processing Plants

In poultry processing plants, disinfectants are often added to pre-chilling water tanks to reduce microbial contamination. The present study aimed at evaluating the effect of five disinfectants (acidified sodium chlorite, alkyl dimethyl benzyl ammonium chloride, chlorine dioxide, peracetic acid, and sodium hypochlorite) on the populations of food quality indicator microorganisms and on Salmonella Enteritidis (SE) in the presence and absence of organic matter. The results showed that chlorine dioxide and sodium hypochlorite did not reduce microbial carcass counts. On the other hand, acidified sodium chlorite, alkyl dimethyl benzyl ammonium chloride and peracetic acid reduced total and fecal coliform counts. Peracetic acid reduced the number of psychrotrophic microorganisms. All products were effective in reducing SE counts only in the absence of organic matter. Acidified sodium chlorite, alkyl dimethyl benzyl ammonium chloride and peracetic acid could be candidates for the replacement of sodium hypochlorite (commonly used in Brazil) in pre-chilling tanks.

Salmonella spp. in meat-type quails (Coturnix coturnix coturnix) in the State of São Paulo, Brazil

In the present study Salmonella spp. was surveyed in four flocks of meat-type quails reared in a farm that also had processing plant on site, located in the region of Bastos, state of São Paulo, Brazil. Meconium samples of one-day-old quail chicks were collected from transport cardboard boxes. Cecal content was collected on days 7, 14, 21, 28 and 35 of rearing. At 36 days of age, birds were slaughtered in the farm's processing plant, where two samples of water from the scalding and the chilling tanks and four carcasses per flock were collected. All samples were examined for Salmonella spp. using traditional bacteriological methods. Salmonella spp. was present in meconium samples of three flocks and in cecal feces of the four flocks. This bacterium was also isolated in the chiller water and in the carcasses of three of the evaluated flocks and in the scalding water of one flock. In this study, S. enterica subspecies enterica 4, 5, 12; S. Corvalis; S. Give; S. Lexington; S. Minnesota; S. Schwarzengrund; S. Rissen and S. Typhimurium were the eight serovars identified.(AU)

Salmonella spp. in meat-type quails (Coturnix coturnix coturnix) in the State of São Paulo, Brazil

In the present study Salmonella spp. was surveyed in four flocks of meat-type quails reared in a farm that also had processing plant on site, located in the region of Bastos, state of São Paulo, Brazil. Meconium samples of one-day-old quail chicks were collected from transport cardboard boxes. Cecal content was collected on days 7, 14, 21, 28 and 35 of rearing. At 36 days of age, birds were slaughtered in the farm's processing plant, where two samples of water from the scalding and the chilling tanks and four carcasses per flock were collected. All samples were examined for Salmonella spp. using traditional bacteriological methods. Salmonella spp. was present in meconium samples of three flocks and in cecal feces of the four flocks. This bacterium was also isolated in the chiller water and in the carcasses of three of the evaluated flocks and in the scalding water of one flock. In this study, S. enterica subspecies enterica 4, 5, 12; S. Corvalis; S. Give; S. Lexington; S. Minnesota; S. Schwarzengrund; S. Rissen and S. Typhimurium were the eight serovars identified.

Índice de patogenicidade, produção de hemolisina e sorogrupo de amostras de Escherichia coli isoladas de aves de postura comercial / Pathogenicity index, production of hemolysin and serogroup of samples of Escherichia coli isolated from commercial laying hens

Este estudo avaliou o índice de patogenicidade, a produção de hemolisina e a determinação de sorogrupos de cepas deEscherichia coli isoladas de fígado de aves de postura comercial com um dia de idade. Para este estudo, foram analisados 32 lotes, dos quais 15 foram positivos para o isolamento de E. coli no fígado, totalizando vinte e quatro amostras. A patogenicidade dos isolados foi determinada por inoculação no saco aéreo de pintinhos e classificada como alta, intermediária, baixa ou não-patogênica. Os sorogrupos foram identificados utilizando um conjunto de antissoros anti-O (O1 a O180). A produção de hemolisina foi determinada por semeadura em ágar sangue de galinha (8%) e em placas de ágar sangue de carneiro (8%). Do total de amostras estudadas, 17 (70,83%) foram classificadas como não patogênica, 6 (25%) como de baixa patogenicidade e 1 (4,17%) de alta patogenicidade. Foram identificados 14 sorogrupos diferentes: O1, O2, O5, O8, O15, O18, O22, O36, O64, O70, O75, O115, O132, O141. Cinco cepas não tiveram o sorogrupo identificado. Com relação ao teste de produção de hemolisina, todas as cepas foram consideradas negativas, tanto para o teste realizado com ágar sangue de galinha quanto para o de carneiro. Os resultados obtidos neste estudo demonstram a importância de se identificar as cepas prevalentes deE. colinas diferentes regiões produtoras, podendo ser utilizados em estudos epidemiológicos.

This work evaluated the index of pathogenicity, the production of hemolysin and determination of serogroups in Escherichia coli strains isolated from liver of commercial laying hens with one day of age. Thirtytwo lots were analyzed, of which 15 were positive for the isolation ofE. coli in the liver, for a total of 24 samples. The pathogenicity in one-day-old chicks was determined by inoculation in air sac and was classified as high, intermediate or low pathogenicity, or non-pathogenic. Serogroups were identified using a set of anti-O antisera (O1 to O180). The production of hemolysin was determined by plating on chicken blood agar (8%) and sheep blood agar (8%). Of the samples studied, 17 (70.83%) were classified as non-pathogenic, 6 (25%) as low pathogenicity and 1 (4.17%) as high pathogenicity. Fourteen different serogroups were identified: O1, O2, O5, O8, O15, O18, O22, O36, O64, O70, O75, O115, O132 and O141, while 5 samples were non-typable. Regarding the test for production of hemolysin, all strains were considered negative for both the test performed with chicken blood agar and that with sheep blood agar. The results of this study demonstrate the importance of identifying the prevalent strains of E. coli in different producing regions, as this information can be used in epidemiological studies.

ÍNDICE DE PATOGENICIDADE, PRODUÇÃO DE HEMOLISINA E SOROGRUPO DE AMOSTRAS DE ESCHERICHIA COLI ISOLADAS DE AVES DE POSTURA COMERCIAL

ABSTRACT This work evaluated the index of pathogenicity, the production of hemolysin and determination of serogroups in Escherichia coli strains isolated from liver of commercial laying hens with one day of age. Thirtytwo lots were analyzed, of which 15 were positive for the isolation ofE. coli in the liver, for a total of 24 samples. The pathogenicity in one-day-old chicks was determined by inoculation in air sac and was classified as high, intermediate or low pathogenicity, or non-pathogenic. Serogroups were identified using a set of anti-O antisera (O1 to O180). The production of hemolysin was determined by plating on chicken blood agar (8%) and sheep blood agar (8%). Of the samples studied, 17 (70.83%) were classified as non-pathogenic, 6 (25%) as low pathogenicity and 1 (4.17%) as high pathogenicity. Fourteen different serogroups were identified: O1, O2, O5, O8, O15, O18, O22, O36, O64, O70, O75, O115, O132 and O141, while 5 samples were non-typable. Regarding the test for production of hemolysin, all strains were considered negative for both the test performed with chicken blood agar and that with sheep blood agar. The results of this study demonstrate the importance of identifying the prevalent strains of E. coli in different producing regions, as this information can be used in epidemiological studies.

RESUMO Este estudo avaliou o índice de patogenicidade, a produção de hemolisina e a determinação de sorogrupos de cepas deEscherichia coli isoladas de fígado de aves de postura comercial com um dia de idade. Para este estudo, foram analisados 32 lotes, dos quais 15 foram positivos para o isolamento de E. coli no fígado, totalizando vinte e quatro amostras. A patogenicidade dos isolados foi determinada por inoculação no saco aéreo de pintinhos e classificada como alta, intermediária, baixa ou não-patogênica. Os sorogrupos foram identificados utilizando um conjunto de antissoros anti-O (O1 a O180). A produção de hemolisina foi determinada por semeadura em ágar sangue de galinha (8%) e em placas de ágar sangue de carneiro (8%). Do total de amostras estudadas, 17 (70,83%) foram classificadas como não patogênica, 6 (25%) como de baixa patogenicidade e 1 (4,17%) de alta patogenicidade. Foram identificados 14 sorogrupos diferentes: O1, O2, O5, O8, O15, O18, O22, O36, O64, O70, O75, O115, O132, O141. Cinco cepas não tiveram o sorogrupo identificado. Com relação ao teste de produção de hemolisina, todas as cepas foram consideradas negativas, tanto para o teste realizado com ágar sangue de galinha quanto para o de carneiro. Os resultados obtidos neste estudo demonstram a importância de se identificar as cepas prevalentes deE. colinas diferentes regiões produtoras, podendo ser utilizados em estudos epidemiológicos.