RESUMO
In this study, a total of 248 ground beef samples were analyzed for the presence of Shiga toxin-producing Escherichia coli (STEC). Out of these samples, only one (0.4%) tested positive for STEC. Further analysis using PCR confirmed the presence of all tested genes associated with STEC, including stx1, stx2, eae, ehx, uid, rfbO157, and fliCH7 in this isolate. Interestingly, no STEC strains were detected in the remaining 100 beef cut samples or the 100 chicken cut samples, indicating the absence of detectable STEC contamination in those specific samples. The isolated strain exhibited significant cytotoxic activity in Vero cells, indicating its ability to produce cytotoxic Shiga toxins. To further investigate the strain, whole-genome sequencing (WGS) analyses were performed. The resistome analysis revealed the absence of acquired antimicrobial resistance genes, indicating a pan-susceptible phenotype. However, this strain presented chromosomal mutations in gyrA, gyrB, parC, parE, pmrA, pmrB, and folP. Plasmid analysis identified the presence of two plasmids, namely IncFIB(AP001918) and IncFII. The multi-locus sequence typing (MLST) identified the strain as belonging to sequence type (ST) 11, which is associated with E. coli O157:H7 strains. The virulome analysis confirmed the presence of several canonical virulence markers, including stx1, stx2, eae-g01-gamma, ehxA, stx1a-O157, and stx2a-O157. Overall, this study identified for the first time a rare occurrence of STEC contamination in ground beef, with the isolated strain belonging to the highly virulent O157:H7 serotype. These findings contribute to our understanding of STEC prevalence and characteristics in food samples, highlighting the importance of effective food safety measures to prevent potential health risks associated with STEC contamination.
RESUMO
Introduction. Shiga toxin-producing Escherichia coli (STEC) is a foodborne pathogen associated with clinical cases of diarrhea in humans. Its main virulence factors are the Shiga toxins (Stx1 and Stx2). Cattle are the main reservoir of STEC, and many outbreaks in humans have been related to the consumption of undercooked ground beef contaminated with this pathogen. Objective. To determine the prevalence of STEC in ground beef commercialized in all the butcher shops of a township in the department of Quindío and to characterize the virulence genes of the strains found. Materials and methods. Thirty ground beef samples were taken in three different times; stx genes and other STEC virulence factors (eae, ehxA, saa) were detected by multiplex PCR. Results. The overall prevalence of STEC was 33.33 % (10/30 positive samples). We isolated eight non-O157 (LEE-negative) strains with four different genetic profiles: stx 2 / stx 2-ehxA-saa / stx 1-stx 2-ehxA-saa / stx 1-saa. Conclusion. This is the first report on the prevalence of STEC in ground beef in a township in the department of Quindío.
Introducción. Escherichia coli productora de toxina Shiga (STEC) es un agente patógeno de origen alimentario asociado a casos clínicos de diarrea en humanos; sus principales factores de virulencia son las toxinas Shiga (Stx1 y Stx2). El principal reservorio de STEC es el ganado bovino y muchos brotes en humanos se han relacionado con el consumo de carne mal cocida contaminada con este agente patógeno. Objetivo. El objetivo de este trabajo fue determinar la prevalencia de STEC en carne molida comercializada en todas las carnicerías de un municipio del departamento del Quindío y caracterizar los genes de virulencia de las cepas encontradas. Materiales y métodos. Se tomaron 30 muestras de carne molida en tres momentos diferentes; se detectaron los genes stx y otros factores de virulencia de STEC (eae, ehxA, saa) mediante PCR Multiplex. Resultados. Los resultados mostraron una prevalencia global de STEC del 33,33 % (10/30 muestras positivas). En total se aislaron ocho cepas STEC no-O157 (LEE-negativas) con cuatro perfiles genéticos diferentes: stx 2 / stx 2-ehxA-saa / stx 1-stx 2-ehxA-saa / stx 1-saa. Conclusión. Este es el primer reporte que muestra la prevalencia de STEC en carne molida en un municipio del departamento del Quindío.
Assuntos
Escherichia coli Shiga Toxigênica , PrevalênciaRESUMO
The aims of the present study were: a) to estimate the minimal dose of gamma irradiation required to reduce 5 log CFU/g of native O157 and non-O157 Shiga toxin-producing Escherichia coli population in ground beef samples inoculated with high inoculum; b) to assess its effectiveness in samples with low inoculum and 3) to evaluate consumer acceptance. Based on the results, 1 kGy was estimated as the minimal dose of gamma irradiation required to reduce 5 log CFU/g of STEC in ground beef. However, when samples with low inoculum level were subjected to 1 kGy, 3.9% of the samples were positive for stx and eae genes after an enrichment step. Consumer acceptance analysis was carried out with samples subjected to 2.5 kGy and no significant differences were found between irradiated and control samples. Therefore, 2.5 kGy was identified as the gama irradiation dose that reduces STEC but has no impact on consumer acceptance of ground beef.
Assuntos
Irradiação de Alimentos/métodos , Produtos da Carne/microbiologia , Escherichia coli Shiga Toxigênica/efeitos da radiação , Animais , Argentina , Bovinos , Comportamento do Consumidor , Raios gama , Humanos , Escherichia coli Shiga Toxigênica/genéticaRESUMO
Atypical enteropathogenic (serotypes O4:H16, O8:H25, O68:H2, O105:H7, and OR:H25) and Shigatoxigenic (ONT:H46) Escherichia coli were isolated from samples of ground beef and poultry breast purchased in Botucatu, Brazil. Phenotypic and molecular characterization indicated the potential of these isolates to adhere to host epithelial cells and cause damage.
Assuntos
Infecções por Escherichia coli/veterinária , Carne/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Brasil , Bovinos , Doenças dos Bovinos/microbiologia , Galinhas , Infecções por Escherichia coli/microbiologia , Contaminação de Alimentos/análise , Contaminação de Alimentos/economia , Carne/economia , Filogenia , Aves Domésticas , Doenças das Aves Domésticas/microbiologia , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genéticaRESUMO
Abstract Ground bovine meat is commonly consumed by the population of Brazil. However, it constitutes an excellent medium for the multiplication of microorganisms due to available nutrients and handling practices prior to consumption. Here, we examined 100 samples of ground beef for the presence of diarrheagenic Escherichia coli (DEC) pathotypes by PCR, and characterized isolates by analyzing their adherence to HEp-2 cells, serotype, antimicrobial susceptibility, and phylogeny. Enteroaggregative E. coli was detected in five (5%) meat samples, Shiga toxin-producing E. coli in three (3%), and atypical enteropathogenic E. coli in two (2%). According to the phylogeny, six isolates (60%) were classified in group A, two (20%) in group B1, and two (20%) in group E. The detected serotypes were O3:H2, O93:H9, O93:H46, O105ab:H7, O152:H8, O156:H10, and O175:H7. The antimicrobial susceptibility testing showed that one sample (10%) was resistant to ampicillin, two (20%) to sulfamethoxazole-trimethoprim, and two (20%) to cephalothin. Based on these results, bovine ground meat for human consumption can serve as a reservoir of DEC, which emphasizes the importance of appropriate hygienic-sanitary conditions during handling at every stage from slaughter to table.
Assuntos
Escherichia coli/isolamento & purificação , Carne Vermelha/microbiologia , Sorotipagem/instrumentação , Gastroenterite/patologiaRESUMO
Shiga toxin-producing Escherichia coli (STEC) is one of the main cause of foodborne disease worldwide, but isolation rates or characteristics of this bacteria from ground beef in Chile are unknown. The present study aimed to isolate and characterize non-O157 STEC from ground beef sold at retail in the city of Santiago, Chile. We analyzed 430 ground beef samples for the presence of STEC, and isolated the microorganism in 10% of samples (43/430). We obtained 56 isolates from the 43 positive samples; 55 of these (98.2%) fermented sorbitol. Most isolates (98.2%; 55/56) showed ß-glucoronidase activity, and only six (10.7%; 6/56) were resistant to tellurite. Among the virulence factors studied (stx1, stx2, eae, and hlyA), stx2 was the only virulence factor in 41% of the isolates (23/56), whereas 10.7% (6/56) of isolates carried a combination of three virulence factors (stx1 + stx2 + hlyA). None of the isolates carried the gene eae. Finally, isolates were neither serogroups O157 nor "big six". In conclusion, ground beef sold in Santiago, Chile is contaminated with STEC; however, further studies are required for understanding their virulence potential.
Assuntos
Carne Vermelha/microbiologia , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Bovinos , Chile , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Contaminação de Alimentos/análise , Carne Vermelha/economia , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/metabolismo , Fatores de Virulência/genética , Fatores de Virulência/metabolismoRESUMO
Abstract Hibiscus sabdariffa L. is used in traditional medicine because of its bioactive properties, such as antioxidant and antibacterial. Escherichia coli is a Gram-negative bacteria and as an indicator of contamination in food. The aim of this work was to evaluate the anti-Escherichia coli effect and the change in pH on the control of aerobic mesophilic microorganisms, using hydroethanolic extract of H. sabdariffa L. in different concentrations in a meat model, verifying its potential as food additive for microbiological stability on ground beef during cooling storage. For the preparation of the treatments, the meat experimental units were elaborated with different concentrations of the vegetal extract (5, 10, 15 and 20%), ground beef and contaminated with E. coli. For pH evaluation, the meat experimental units were added different percentages of hydroethanolic extract. The H. sabdariffa L. antibacterial action reduced two logarithmic levels in practically all treatments. The best pH result was obtained in the meat containing 30% of the extract. The hydroethanolic extracts of Hibiscus sabdariffa L. showed anti-Escherichia coli activity in the presence of refrigerated ground beef. Analyzing the pH results and the count of aerobic mesophilic bacteria, it is possible this extract to be used as a natural food additive.
RESUMO
Shiga toxin-producing Escherichia coli (STEC) are recognized as food-borne pathogens. We developed and validated two SYBR green PCR (SYBR-PCR) and a real-time multiplex PCR (RT-PCR) to detect stx1 and stx2 genes in meat samples, and compared these techniques in ground beef samples from retail stores. One set of primers and one hydrolysis probe were designed for each stx gene. For RT-PCR, an internal amplification control (IAC) was used. All PCR intra-laboratory validations were performed using pure strains and artificially contaminated ground beef samples. A total of 50 STEC and 30 non-STEC strains were used. Naturally contaminated ground beef samples (n=103) were obtained from retail stores and screened with SYBR-PCR and RT-PCR, and stx-positive samples were processed for STEC isolation. In the intra-laboratory validation, each PCR obtained a 1×10(2) CFU mL(-1) limit of detection and 100% inclusivity and exclusivity. The same results were obtained when different laboratory analysts in alternate days performed the assay. The level of agreement obtained with SYBR-PCR and RT-PCR was kappa=0.758 and 0.801 (P<0.001) for stx1 and stx2 gene detection, respectively. Two PCR strategies were developed and validated, and excellent performance with artificially contaminated ground beef samples was obtained. However, the efforts made to isolate STEC from retail store samples were not enough. Only 11 STEC strains were isolated from 35 stx-positive ground beef samples identically detected by all PCRs. The combination of molecular approaches based on the identification of a virulence genotypic profile of STEC must be considered to improve isolation.
Assuntos
Contaminação de Alimentos/análise , Carne/microbiologia , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Escherichia coli Shiga Toxigênica/isolamento & purificação , Animais , Benzotiazóis , Bovinos , Primers do DNA/genética , Diaminas , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Carne/análise , Reação em Cadeia da Polimerase Multiplex/instrumentação , Compostos Orgânicos/metabolismo , Quinolinas , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Toxina Shiga/genética , Toxina Shiga/metabolismo , Escherichia coli Shiga Toxigênica/genética , Escherichia coli Shiga Toxigênica/metabolismoRESUMO
Meat and particularly ground beef is frequently associated with Food Poisoning episodes and breeches in Food Safety. The main goal of this research was to evaluate the bactericide effect of the probiotic Enterococcus faecalis CECT7121, against different pathogens as: Escherichia coli O157:H7, Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes, inoculated in ground beef meat. Three studies were performed to evaluate the inhibition of E. faecalis CECT7121 on ground beef meat samples inoculated with pathogens: Study I: Samples (100 g meat) were inoculated with pathogens (10(3) CFU/g)) and E. faecalis CECT7121 (10(4) CFU/g) simultaneously. Study II: Samples were inoculated with E. faecalis CECT7121 24 h before the pathogens. Study III: E. faecalis CECT7121were inoculated 24 h after pathogens. The viable counts were performed at 0, 24, 48 and 72 h post-inoculation. The simultaneous inoculation of E. faecalis CECT7121 with E. coli O157:H7 strains resulted in the absence of viable counts of bacteria at 72 h post-treatment. However, when the probiotic was added 24 h before and 24 h after the pathogen E. coli O157:H7, viable cells were not detected at 24 h and 48 h post-treatment, respectively. Consistently, neither S. aureus nor Cl. perfringens viable bacteria were detected at 48 h in whole assays when inoculated with E. faecalis CECT7121. The same trend than described before was obtained after applying the 3 models assayed for L. monocytogenes. The current assays demonstrated the bactericide activity of E. faecalis CECT7121 strain on bacterial pathogens in ground beef meat.
RESUMO
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.
RESUMO
El objetivo del estudio fue aislar y caracterizar E. coli O157:H7 a partir de carne molida fresca de bovino obtenida en diferentes mercados de abastos. Se analizaron 195 muestras; para el aislamiento y enumeración de E. coli se utilizó la técnica del Numero Más probable mediante tubos múltiples; para el aislamiento y caracterización de E. coli O157:H7 el enriquecimiento selectivo y el análisis bioquímico, las colonias características se confirmaron mediante pruebas serológicas. Para determinar la presencia de shigatoxina (stx1, stx2) e intimina (eae A) se empleó la técnica de PCR multiplex en tiempo real y para enterohemolisina la prueba de hemólisis. El 87.18% de la muestras fue positivo para E. coli y el 77.95% presentó un recuento igual o superior a 50 NMP/g. Se obtuvieron 3 (1.54%) cepas de E. coli O157:H7, una stx1 +/ stx2 +/ eaeA - y enterohemolisina -, una stx 1 +/ stx2 -/ eaeA + y enterohemolisina - y la otra stx1 -/ stx2 -/ eaeA - y enterohemolisina +. También se obtuvieron 4 cepas (2.05%) de E. coli O157 no H7, ninguna presentó factores de virulencia. El estudio reveló el riesgo potencial de que E. coli O157:H7 afecte a la población de Lima
The aim of the study was to isolate and characterize E. coli O157: H7 from fresh ground beef obtained in different food markets. 195 samples were analyzed, for isolation and enumeration of E. coli was used most probable number technique using multiple tubes, for the isolation and characterization of E. coli O157: H7 selective enrichment and biochemical analysis, characteristic colonies were confirmed serologically. To determine the presence of shigatoxin (stx1, stx2) and intimin (eaeA) was done with multiplex real time PCR and for enterohemolysin the hemolysis test. The 87.18% of the samples were positive for E. coli and 77.95% had a count greater than or equal to 50 NMP/g. Were obtained 3 (1.54%) strains of E. coli O157: H7, one stx1 +/ stx2 +/eaeA - and enterohemolysin -, one stx1 +/stx2 -/eaeA + and enterohemolysin - and the other stx1 -/stx2 -/eaeA - and enterohemolysin +. Also obtained 4 (2.05%) strains of E. coli O157 non H7, no virulence factors presented. The study found that the risk potential E. coli O157: H7 affecting the population of Lima
RESUMO
Meat and particularly ground beef is frequently associated with Food Poisoning episodes and breeches in Food Safety. The main goal of this research was to evaluate the bactericide effect of the probiotic Enterococcus faecalis CECT7121, against different pathogens as: Escherichia coli O157:H7, Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes, inoculated in ground beef meat. Three studies were performed to evaluate the inhibition of E. faecalis CECT7121 on ground beef meat samples inoculated with pathogens: Study I: Samples (100 g meat) were inoculated with pathogens (10³ CFU/g)) and E. faecalis CECT7121 (10(4) CFU/g) simultaneously. Study II: Samples were inoculated with E. faecalis CECT7121 24 h before the pathogens. Study III: E. faecalis CECT7121were inoculated 24 h after pathogens. The viable counts were performed at 0, 24, 48 and 72 h post-inoculation. The simultaneous inoculation of E. faecalis CECT7121 with E. coli O157:H7 strains resulted in the absence of viable counts of bacteria at 72 h post-treatment. However, when the probiotic was added 24 h before and 24 h after the pathogen E. coli O157:H7, viable cells were not detected at 24 h and 48 h post-treatment, respectively. Consistently, neither S. aureus nor Cl. perfringens viable bacteria were detected at 48 h in whole assays when inoculated with E. faecalis CECT7121. The same trend than described before was obtained after applying the 3 models assayed for L. monocytogenes. The current assays demonstrated the bactericide activity of E. faecalis CECT7121 strain on bacterial pathogens in ground beef meat.
Assuntos
Humanos , Animais , Criança , Bovinos , Infecções Bacterianas , Carne/análise , Enterococcus faecalis/isolamento & purificação , Conservação de Alimentos , Probióticos/isolamento & purificação , Produtos da Carne/análise , Bovinos , Microbiologia de Alimentos , Amostras de Alimentos , Métodos , VirulênciaRESUMO
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.
RESUMO
Meat and particularly ground beef is frequently associated with Food Poisoning episodes and breeches in Food Safety. The main goal of this research was to evaluate the bactericide effect of the probiotic Enterococcus faecalis CECT7121, against different pathogens as: Escherichia coli O157:H7, Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes, inoculated in ground beef meat. Three studies were performed to evaluate the inhibition of E. faecalis CECT7121 on ground beef meat samples inoculated with pathogens: Study I: Samples (100 g meat) were inoculated with pathogens (10³ CFU/g)) and E. faecalis CECT7121 (10(4) CFU/g) simultaneously. Study II: Samples were inoculated with E. faecalis CECT7121 24 h before the pathogens. Study III: E. faecalis CECT7121were inoculated 24 h after pathogens. The viable counts were performed at 0, 24, 48 and 72 h post-inoculation. The simultaneous inoculation of E. faecalis CECT7121 with E. coli O157:H7 strains resulted in the absence of viable counts of bacteria at 72 h post-treatment. However, when the probiotic was added 24 h before and 24 h after the pathogen E. coli O157:H7, viable cells were not detected at 24 h and 48 h post-treatment, respectively. Consistently, neither S. aureus nor Cl. perfringens viable bacteria were detected at 48 h in whole assays when inoculated with E. faecalis CECT7121. The same trend than described before was obtained after applying the 3 models assayed for L. monocytogenes. The current assays demonstrated the bactericide activity of E. faecalis CECT7121 strain on bacterial pathogens in ground beef meat.(AU)
Assuntos
Intoxicação , Enterococcus/patogenicidade , Escherichia/patogenicidade , Staphylococcus/patogenicidade , Carne/análise , ProbióticosRESUMO
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.
RESUMO
Este estudo objetivou averiguar a qualidade da carne moída comercializada em 10 estabelecimentos, açougues e supermercados, na cidade de Barra do Garças-MT, quanto a presença de Salmonella spp., e ocorrência de bactérias aeróbias mesófilas, coliformes totais, Escherichia coli, Staphylococcus aureus e fungos (bolores e leveduras). Para tanto, coletou-se três amostras por estabelecimento perfazendo um total de 30, que foram analisadas pelos métodos: tradicional para determinação da presença de Samonellaspp. e pelo método rápido, com placas Petrifilm TM 3M, para os demais microrganismos pesquisados. Destas 30 amostras, cinco (17%) foram positivas para a presença de Salmonellaspp., A contagem de S. aureus em 20% das amostras variaram de 2,7 x 102 UFC/g a 9,3 x 102 UFC/g.E. colifoi verificada em contagens de 1,0 x 101 UFC/g até 6,5 x 102 UFC/g. Encontrou-se Coliformes totais em 13,3% das amostras, com contagens entre 1,2 x 103 UFC/g a 4,4 x 104 UFC/g. As bactérias aeróbias mesófilas nas amostras variaram de 1,3 x 103 UFC/g a 2,7 x 105 UFC/g, e observou-se contagens de bolores e leveduras de 1,1 x 102 UFC/g a 4,4 x 104 UFC/g. A presença de cepas de Salmonellaspp. e S. aureus nas amostras de carne moída demonstram falha higiênica na manipulação desta, e representando risco a saúde dos consumidores.
This study aimed to investigate ground beef sold in 10 establishments, like as butcher shops and supermarkets, in the town of Barra do Garças-MT, for presence of Salmonella spp.,. and occurrence of mesophilic aerobic bacteria, coliforms, Escherichia coli and Staphylococcus aureus. For this, three samples were collected in the places of sale for a total of 30 samples which were analyzed by the methods: traditional (Salmonellaspp.) and fast using Petrifilm TM 3M plates (others microorganisms). Of these 30 samples, five (17%) were positive for the presence of Salmonellaspp., and 20% for S. aureuscounts ranging from 2,7 x 102 CFU/g to 9,3 x 102 CFU/g, and 100% with 1,0 x 101 CFU/g to 6,5 x 102 CFU/g in E. coli. Total coliforms were found in 13, 3% of the samples with counts between 1,2 x 103 CFU/gto 4,4 x 104 CFU/g, and yeast and mold counts between 1,1 x 102 CFU/g to 4,4 x 104 CFU/g and mesophilic aerobic bacteria between 1,3 x 103 to 2,7 x 104 CFU/g. The presence of Salmonellaspp. strains and S. aureusin ground beef samples showed fault in hygienic handling of these establishments, and still represent a health risk for consumers.
Assuntos
Contaminação de Alimentos , Perfis Sanitários , Padrão de Identidade e Qualidade para Produtos e Serviços , Carne/microbiologiaRESUMO
A carne moída é um produto que todo consumidor conhece, porém existe outra opção, a carne moída homogeneizada, que apesar de estar no mercado há mais de 20 anos, não é ainda conhecida, tanto na forma da sua obtenção, quanto sobre sua qualidade. Este trabalho teve como objetivo descrever como é a produção deste produto, bem como avalia a qualidade através de indicadores microbiológicos. A metodologia aplicada foi a colheita de 325 amostras de carne moída homogeneizada, em oitenta e três lojas, de uma rede de supermercados, escolhidas aleatoriamente, em nove estados da federação, no período de novembro de 2009 a abril de 2011. Os resultados foram avaliados segundo a RDC 12 da ANVISA verificando se as amostras estavam ou não dentro do padrão legal.(AU)
The ground beef is a product thatevery consumer knows, but there isanother option, the homogenizedground beef, which despite being inthe market for over 20 years, is notyet known, both in the form of obtainingit. as about its quality. Thisstudy had as targets to describehow to produce this type of productas well as assess the microbiologicalquality by means of indicators.The utilized methodology was thecollection of 325 samples of homogenizedground beef in 83 randomlyselected stores of a supermarketchain located in nine states of theBrazilian federation, held from November2009 to April 2011. The resultswere evaluated according tothe RDC ANVISA 12 to verify whetherthe analyzed samples were inthe legal standards.(AU)
Assuntos
Animais , Bovinos , Produção de Alimentos , Carne/microbiologia , Carne/análise , Conservação de Alimentos , Armazenamento de Alimentos , Normas de Qualidade de AlimentosRESUMO
Shiga toxin-producing Escherichia coli (STEC) are foodborne pathogens that cause mild or serious diseases and can lead to people death. This study reports the prevalence and characteristics of STEC O157 and non-O157 in commercial ground beef and environmental samples, including meat table, knife, meat mincing machine, and manipulator hands (n = 450) obtained from 90 retail markets over a nine-month period. The STEC isolates were serotyped and virulence genes as stx (Shiga toxin), rfb(O157)] (O157 lipopolysaccharide), fliC(H7) (H7 flagellin), eae (intimin), ehxA (enterohemolysin) and saa (STEC autoagglutinating adhesin), were determined. STEC O157 were identified in 23 (25.5%) beef samples and 16 (4.4%) environmental samples, while STEC non-O157 were present in 47 (52.2%) and 182 (50.5%), respectively. Among 54 strains isolated, 17 were STEC O157:H7 and 37 were STEC non-O157. The prevalent genotype for O157 was stx(2)/eae/ehxA/fliC(H7) (83.4%), and for STEC non-O157 the most frequent ones were stx(1)/stx(2)/saa/ehxA (29.7%); stx(2) (29.7%); and stx(2)/saa/ehxA (27%). None of the STEC non-O157 strains were eae-positive. Besides O157:H7, other 20 different serotypes were identified, being O8:H19, O178:H19, and O174:H28 the prevalent. Strains belonging to the same serotype could be isolated from different sources of the same retail market. Also, the same serotype could be detected in different stores. In conclusion, screening techniques are increasingly sensitive, but the isolation of STEC non-O157 is still a challenge. Moreover, with the results obtained from the present work, although more studies are needed, cross-contamination between meat and the environment could be suspected.
Assuntos
Microbiologia Ambiental , Carne/microbiologia , Escherichia coli Shiga Toxigênica/classificação , Escherichia coli Shiga Toxigênica/isolamento & purificação , Argentina , Flagelina/genética , Contaminação de Alimentos , Indústria Alimentícia , Genótipo , Humanos , Epidemiologia Molecular , Antígenos O/genética , Prevalência , Sorotipagem , Escherichia coli Shiga Toxigênica/genética , Fatores de Virulência/genéticaRESUMO
Background: Ground meat stands among the most consumed meat products, being its safety easily damaged by increasing the surface area of contact after grinding it, what facilitates the growth of pathogenic and deteriorating micro-organisms. In order to decrease the lost because of deterioration, trade has been utilizing fraudulent devices as the addition of intentional preservative such as sodium sulfite. This research aimed to identify the presence of sodium sulfite in cooled ground beef commercialized in business establishments located in Porto Alegre city, state of Rio Grande do Sul, Brazil. In addition, it addressed the evaluation of the influence of using this addictive in cooled ground meat front its physic-chemical and microbiological characteristics and residual capacity in the first 48 hours.Materials, Methods & Results: A total of 55 samples of cooled ground beef, from non-noble cuts, acquired in business establishments located in Porto Alegre city, were collected. Thus, microbiological analyses for total and thermotolerant coliphorms, Clostridium Sulfite reducers, Staphylococcus coagulase-positive, Salmonella sp. and a qualitative test, for verifying the presence of sodium sulfite, were carried out. Subsequently, half of the negative samples (24) in the sulfite test were separated, and 0.04 g of this addictive for each 50 g of ground beef (0.08%) was, then, added.
Background: Ground meat stands among the most consumed meat products, being its safety easily damaged by increasing the surface area of contact after grinding it, what facilitates the growth of pathogenic and deteriorating micro-organisms. In order to decrease the lost because of deterioration, trade has been utilizing fraudulent devices as the addition of intentional preservative such as sodium sulfite. This research aimed to identify the presence of sodium sulfite in cooled ground beef commercialized in business establishments located in Porto Alegre city, state of Rio Grande do Sul, Brazil. In addition, it addressed the evaluation of the influence of using this addictive in cooled ground meat front its physic-chemical and microbiological characteristics and residual capacity in the first 48 hours.Materials, Methods & Results: A total of 55 samples of cooled ground beef, from non-noble cuts, acquired in business establishments located in Porto Alegre city, were collected. Thus, microbiological analyses for total and thermotolerant coliphorms, Clostridium Sulfite reducers, Staphylococcus coagulase-positive, Salmonella sp. and a qualitative test, for verifying the presence of sodium sulfite, were carried out. Subsequently, half of the negative samples (24) in the sulfite test were separated, and 0.04 g of this addictive for each 50 g of ground beef (0.08%) was, then, added.
RESUMO
Background: Ground meat stands among the most consumed meat products, being its safety easily damaged by increasing the surface area of contact after grinding it, what facilitates the growth of pathogenic and deteriorating micro-organisms. In order to decrease the lost because of deterioration, trade has been utilizing fraudulent devices as the addition of intentional preservative such as sodium sulfite. This research aimed to identify the presence of sodium sulfite in cooled ground beef commercialized in business establishments located in Porto Alegre city, state of Rio Grande do Sul, Brazil. In addition, it addressed the evaluation of the influence of using this addictive in cooled ground meat front its physic-chemical and microbiological characteristics and residual capacity in the first 48 hours. Materials, Methods & Results: A total of 55 samples of cooled ground beef, from non-noble cuts, acquired in business establishments located in Porto Alegre city, were collected. Thus, microbiological analyses for total and thermotolerant coliphorms, Clostridium Sulfite reducers, Staphylococcus coagulase-positive, Salmonella sp. and a qualitative test, for verifying the presence of sodium sulfite, were carried out. Subsequently, half of the negative samples (24) in the sulfite test were separated, and 0.04 g of this addictive for each 50 g of ground beef (0.08%) was, then, added. In these samples, microbiological and color analyses, and the confirmation of the presence of sodium sulfite, at times 0, 24 and 48, after the simulation of fraud of the samples, were realized. Moreover, for the verification of safety, microbiological analyses for Salmonella sp., Staphylococcus coagulase-positive, Clostridium Sulfite reducers, total and thermo tolerant coliphorms were carried out. Data were analyzed by Analysis of Variance (ANOVA), with pairwise comparisons done by the Tukey test, for a level of significance of 5% (P < 0.05). Thereby, from the total of 55 (100%) analyzed samples of cooled ground beef, two (3.63%) showed the presence of sodium sulfite addictive, and the performed microbiological analyses indicated the presence of Salmonella sp. in eight (14.54%) samples. The values of L*, observed in the samples added with sulfite, maintained constant at times (zero, 24 and 48) compared with the control samples (without addition of sulfite) (42.92; 40.35 and 41.69, vs 44.67; 37.65 and 32.38, respectively), which demonstrated lost of luminosity in the course of time. Also, the values of a* and b* demonstrated stable at times (zero, 24 and 48) in the samples added with addictive (5.72; 6.89; 5.47 and 9.73; 10.69; 9.56, respectively), although this has not occurred with the control samples that presented a gradual lost of these tonalities (11.48; 7.21; 5.91 and 16.11; 12.88; 7.84, respectively) (P < 0.05). Discussion: The results indicated that two (3.63%) samples showed the presence of sodium sulfite, being in disagreement with the Ordinance n. 1004 that forbids the addition of addictive a in natura meat. The presence of Salmonella sp. in samples of ground meat contradicts the Resolution RDC n. 12 that advocates its absence in 25 g of meat products. Furthermore, the conducted analyses of color proved that the addition of addictive in meat maintains its fresh color and appearance, beyond inhibiting reactions of enzymatic and non-enzymatic darkness during processing and storage. By considering the obtained results, it may be concluded that despite of a low incidence of fraud for finding sodium sulfite, this practice exists in business establishments located in Porto Alegre city, state of Rio Grande do Sul, Brazil, even though it can be concealed for the lack of continuous studies and legal analytical inspections, endangering sanitary-hygienic conditions of the products.