Biofilm formation of Staphylococcus aureus from milk and expression of the adhesion genes ebpS and cna at different temperatures.

This study investigated the ability of Staphylococcus aureus isolates from milk to form biofilm, through detection of adhesion genes, investigating exopolysaccharide (EPS) production and biofilm formation on polystyrene (PS) and stainless steel (SS) surfaces, and by quantifying the expression of ebpS and cna genes under different temperatures and culture media. Among the 31 isolates, the adhesion genes ebpS and cna were found in 81% and 61% of the isolates, respectively. The screening tests for phenotype revealed that 58% of the isolates were EPS producers, and 45% showed the ability to produce biofilm on PS. Nine of the 31 isolates were selected to verify their ability to form biofilm on SS, of which 3 were non-biofilm producers, 3 were poor biofilm producers, and 3 were moderate biofilm producers. However, all nine isolates produced biofilm on SS, regardless of their phenotypic profile on PS. Reverse-transcriptase quantitative PCR (RT-qPCR) revealed no variation in the expression levels of ebpS and cna genes at different temperatures, except for isolate S24 at 10 °C, for both genes tested. Moreover, RT-qPCR assays revealed that the expression levels of the adhesion genes ebpS and cna are isolate- and temperature-dependent; however, they are independent of the phenotypic biofilm-formation profile.

Bacteriocin-like substances of Lactobacillus curvatus P99: characterization and application in biodegradable films for control of Listeria monocytogenes in cheese.

The aim of this study was to evaluate the effectiveness of a biodegradable film, with antimicrobial metabolites produced by Lactobacillus curvatus P99 incorporated, targeting the control of Listeria monocytogenes in sliced "Prato" cheese. Tests were performed to evaluate the spectrum of action of cell-free supernatant (CFS) of P99 against different microorganisms, as well as to detect the minimum inhibitory (MIC) and bactericidal (MBC) concentrations against L. monocytogenes Scott A. The detection of genes that encode for the production of bacteriocins and evaluation of their expression were performed. Antimicrobial films were prepared, followed by in vitro and in situ analysis. The MIC and MBC of CFS against L. monocytogenes Scott A was 15.6 µL/mL and 62.5 µL/mL, respectively. Lactobacillus curvatus P99 presented two genes coding for the bacteriocins, which were expressed. Films with added MBC showed activity against different indicator microorganisms and were able to control L. monocytogenes Scott A when used in sliced "Prato" cheese.

Pichia pastoris X-33 has probiotic properties with remarkable antibacterial activity against Salmonella Typhimurium.

Probiotics are live microorganisms which are beneficial for the host when ingested at high enough concentrations. The methylotrophic yeast Pichia pastoris is widely used as heterologous protein production platform. However, its use as probiotic is poorly studied. The objective of this study was to evaluate some probiotic properties of the P. pastoris strain X-33 wild type. The resistance to in vitro and in vivo gastrointestinal conditions, stability in feed, safety, and antibacterial activity against Salmonella Typhimurium were evaluated. The yeast remained viable and persisted at appropriate concentration in the diet for at least 2 months, survived the stresses of the gastrointestinal tract in vitro and in vivo, caused no behavioral changes or lesions when administered to mice, inhibited the growth of S. Typhimurium in culture media, and reduced adhesion of the bacteria to the intestinal cells HCT-116. In the challenge experiment with a LD50 of virulent S. Typhimurium strain, mice supplemented with the yeast had a higher survival rate (50 % when administered by gavage and 80 % via the diet, compared with 20 and 50 %, respectively, in the control group). In addition, the S. Typhimurium concentration in the intestine of the surviving mice was lower; the score of intestinal lesions, lower; and the pathogen, not detected in the liver, spleen, and feces when compared to the control group (p < 0.05). It was concluded that the yeast Pichia pastoris X-33 has probiotic properties with remarkable antibacterial activity against S. Typhimurium.

Microbiological Testing for the Proper Assessment of the Hygiene Status of Beef Carcasses.

Microbiological testing is an important quality management tool in the food industry. In this study, the hygiene status of beef carcasses sampled in eight Brazilian slaughterhouses was assessed by enumeration of different hygiene indicator microorganisms, and a model to establish potential associations among these counts was proposed. The carcasses (n = 464) were surface sampled at four slaughtering steps (step 1: Hide after bleeding; step 2: Carcass after hide removal; step 3: Carcass after evisceration; step 4: Carcass after end washing) and subjected to a counting of mesophilic aerobes (MA), Enterobacteriaceae (EB), total coliforms (TC), and Escherichia coli (EC) using Petrifilm™ plates. Among the sampled beef carcasses (step 4), 32 (6.9%) and 71 (15.3%) presented counts above the microbiological criteria established by (EC) No. 1441/2007 for MA and EB, respectively. Thus, indicating that improvements in slaughter hygiene and a review of process controls are demanded in some of the studied slaughterhouses. The log count differences of EC, TC, and EB from MA were considered as response variables as a function of the slaughtering steps. Differential log counts changed consistently with the steps. The measurements, including the patterns in their inherently random variability, were fairly predictable from steps 1 and 4. The results indicated that differential log counts for TC and EC are not relevant, as their concentrations and random pattern can be inferred from counts of MA and EB. The proposed model can be used as a valuable tool for the design and adoption of feasible quality control programs in beef industries. The adoption of such a tool should have a positive contribution on consumers' health and enhance product quality.

Food isolate Listeria monocytogenes harboring tetM gene plasmid-mediated exchangeable to Enterococcus faecalis on the surface of processed cheese.

The genetic basis of tetracycline resistance in a food isolate Listeria monocytogenes (Lm16) was evaluated. Resistance to tetracycline was associated with the presence of the tetM gene in plasmid DNA. The sequence of tetM showed 100% of similarity with the Enterococcus faecalis sequences found in the EMBL database, suggesting that Lm16 received this gene from E. faecalis. Various size bands were detected in the DNA plasmid analysis, the largest being approximately 54.38 kb. Transferability of the tetM gene was achieved in vitro by agar matings between Lm16 and E. faecalis JH2-2, proving the potential for the spread of tetM by horizontal gene transfer. Furthermore, the conjugation experiments were performed on the surface of processed cheese, confirming the transferability in a food matrix. PCR assays were used to confirm the identity of E. faecalis and to detect the tetM gene in transconjugant bacteria. Additionally, the minimal inhibitory concentration for tetracycline and rifampicin and plasmid profiling were performed. This is the first report of a food isolate L. monocytogenes carrying the tetM gene in plasmid DNA, and it highlights the potential risk of spreading antimicrobial resistance genes between different bacteria.

Occurrence and phenotypic and molecular characterization of Listeriamonocytogenes and Salmonella spp. in slaughterhouses in southern Brazil.

This study addressed the occurrence of Listeriamonocytogenes and Salmonella spp. in bovine carcasses at two slaughterhouses in southern Brazil. Then, the antimicrobial susceptibility profile and the virulence potential of the isolates were evaluated. Two hundred carcasses were sampled at four steps of the slaughter process, with L. monocytogenes being isolated in 12 and Salmonella spp. in 17 carcasses. All L. monocytogenes isolates carried the hlyA, prfA, plcA, plcB, actA, iap, mpl, inlA, inlB, inlC, and inlJ genes, while Salmonella spp. carried invA and hilA. Among the L. monocytogenes isolates, all of them presented virulence determinants and one showed multi-drug resistance. In relationship to Salmonella spp. isolates, many serogroups frequently related to outbreaks of foodborne diseases were identified and four isolates showed resistance to more than one antimicrobial agent. This data highlights the importance of a rigid hygienic-sanitary control during the slaughter process to reduce the risk of cross-contamination and lower the consumer exposure to L. monocytogenes and Salmonella spp. infections.

Virulence and antimicrobial resistance of Salmonella spp. and Escherichia coli in the beef jerky production line.

Intense manipulation during beef jerky production increases the possibility of contamination with pathogenic microorganisms. This study evaluated the contamination by thermotolerant coliforms, Escherichia coli and Salmonella spp., on processing surfaces and raw materials during beef jerky production, as well as in the final product. Thermotolerant coliforms were found on all surfaces tested and in the raw material. Escherichia coli was identified in 6.7% of the surface samples, while Salmonella spp. was found in 3.3% of the surface samples and 8.6% of raw material samples. Virulence genes were detected in Salmonella spp. isolates. One Salmonella spp. isolate was resistant to sulfonamide, while one E. coli isolate was multiresistant, including the presence of resistance genes sul2, strA, strB, tetA and tetB. The presence of coliforms demonstrates failings in hygienic-sanitary procedures. The presence of pathogenic microorganisms causing foodborne diseases in the production line indicates persistent contamination in the production plant. Although the drying process applied to beef jerky should guarantee the safety of the final product, the presence of multiresistant pathogenic microorganisms, presenting virulence genes, should be a matter of concern. Because beef jerky is a ready-to-eat product, a failure in the production process may cause such microorganisms to pose a public health risk.

Resistência antimicrobiana em isolados de Listeria monocytogenes provenientes de alimentos e ambientes de processamento de alimentos / Antimicrobial resistance in Listeria monocytogenes isolates from food and food processing environments

Diversos estudos relatam distintos níveis de resistência antimicrobiana entre os isolados de L. monocytogenes, o que pode ter implicações no tratamento de listeriose. Este estudo avaliou o perfil de resistência de 30 isolados de L. monocytogenes provenientes de alimentos e ambientes de processamento de alimentos do sul do Brasil. Todos os isolados apresentaram resistência ao ácido nalidíxico e a cefoxitina, além de ter sido observada alta ocorrência de isolados resistentes à clindamicina. Para alguns antimicrobianos (rifampicina, tetraciclina, eritromicina, estreptomicina, meropenem, trimetoprim-sulfametoxazol) foi detectado baixo percentual de resistência e, para os demais antimicrobianos testados, não foi observada resistência. Como conclusão, apesar de ter sido observado perfil de resistência para alguns antimicrobianos, os isolados foram suscetíveis aos antimicrobianos comumente utilizados no tratamento de listeriose.

Ação in vivo de Lactococcus lactis subsp. lactis com potencial probiótico na estabilização de células cancerígenas no epitélio colorretal / In vivo action of Lactococcus lactis subsp. lactis with probiotic potential in the stabilization of cancer cells in the colorectal epithelium

Na última década, a ciência contribuiu significativamente para inúmeros avanços em relação ao tratamento e prevenção do câncer colorretal (CCR), porém, a prevalência global e a taxa de mortalidade permanecem elevadas. Há relatos sobre efeitos benéficos de espécies de Bifidobacterium e Lactobacillus com potencial probiótico na prevenção de CCR. No entanto, a bactéria probiótica Lactococcus lactis subps. lactis é comumente utilizada para fins industriais, não havendo comprovações in vivo sobre seu potencial anticarcinogênico. Visto o interesse emergente dos efeitos benéficos dos probióticos a fim de prevenir ou tratar o CCR, o presente estudo objetivou explorar os efeitos de L. lactis subsp. lactis sobre o CCR. Ratos Wistar receberam doses subcutâneas de 1,2 dimetilhidrazina (DMH) e suspensão de L. lactis subsp. lactis por via oral. Após 20 semanas, os tecidos intestinais foram analisados e de acordo com o resultado, o isolado demonstrou potencial anticarcinogênico contra CCR.

Multivariate optimization of Staphylococcus xylosus AD1biomass production using sugarcane molasses plus yeast extract and soybean meal

Staphylococcus xylosusis a microorganism that has important physiological and technological characteristics that make it suitable for use as a starter culture in fermented meat products. For the development of these products in the food industry, it is necessary to produce biomass by the multiplication of starter cultures using low-cost media. This study developed a culture medium based on sugarcane molasses (SCM) supplemented with yeast extract (YE) and soybean meal (SM) to produce S. xylosusAD1 biomass employing a Box Behnken multivariateoptimization design,usingthe best concentrations of the constituents of the culture medium for S. xylosusAD1 growth. By combining the mathematical models by the desirability function, it was possible to establish the optimal condition for the maximum production of viable cells and biomass. The optimal experimental condition was found when the fermentative process medium was composed of 10% SCM, 2% YE and 4% SM. In addition, the results of all experiments, except for the medium formulated with only SCM,presented a better performance than the commercial medium Brain Heart Infusion for the growth of S. xylosusAD1. The culture medium with agro-industrial byproduct (SCM) supplemented with YE and SM is an excellent alternative for producing S. xylosusAD1 biomass.

Microbiological quality and safety assessment in the production of moderate and high humidity cheeses / Qualidade microbiológica e avaliação da segurança na produção de queijos de média e alta umidade

ABSTRACT: Moderate and high humidity cheeses are described as important vehicles of pathogens in many foodborne diseases outbreaks. Microbial contamination can occur in raw material or in the different steps of the product processing due to inadequate hygiene practices. Thus, the aim of this study was to evaluate the microbiological quality and safety in the production of moderate and high humidity cheese. Samples from raw milk, handlers' hands surface, final product were collected in three cheese manufacturing plants located in southern Brazil, with different levels of sanitary control. Effectiveness of milk pasteurization was also evaluated. Thermotolerant coliforms, coagulase-positive staphylococci (CPS), Salmonella spp., and Listeria monocytogenes were evaluated. Raw milk samples showed the highest contamination levels, with enumeration of 1.1x105 most probable number (MPN) mL-1 for thermotolerant coliforms, 4x105 colony-forming units (CFU) mL-1 for CPS and presence of Salmonella spp. CPS were also reported in one sample of handler's hands surface. However, only one sample of the final product was out of Brazilian regulatory standards, exceeding the limit allowed for CPS. Milk pasteurization process used in cheese preparation was effective, regardless the level of sanitary control of the industries. Results highlighted the need for better hygiene practices, in obtaining the raw milk and in the handling during the cheese manufacturing steps.

RESUMO: Os queijos com média e alta umidade são alimentos prontos para o consumo, que têm sido descritos como veiculadores de patógenos em diversos surtos de doenças transmitidas por alimentos. A contaminação microbiana pode ter origem na matéria prima, ou ocorrer durante as etapas de elaboração do produto, através de práticas inadequadas de higiene. Dessa forma, o objetivo deste estudo foi avaliar a qualidade e a segurança microbiológica na produção de queijos de média umidade. Amostras da matéria prima, dos manipuladores e do produto final foram coletadas em três laticínios situados na região sul do Rio Grande do Sul, com diferentes níveis de inspeção sanitária. A eficiência da pasteurização do leite também foi avaliada. Coliformes termotolerantes, estafilococos coagulase-positivos (ECP), Salmonella spp. e Listeria monocytogenes foram avaliados. As amostras de leite cru foram as que apresentaram os maiores níveis de contaminação, com enumeração de 1,1x105 número mais provável (NMP) mL-1 para coliformes termotolerantes, 4x105 unidades formadoras de colônia (UFC) mL-1 para ECP e a presença de Salmonella spp.. Contudo, apenas uma amostra de produto final estava em desacordo com o padrão regulamentar vigente, excedendo o limite permitido para ECP. A pasteurização do leite utilizado no preparo dos queijos foi eficiente em todos os laticínios, independentemente do nível de inspeção sanitária dos estabelecimentos. No entanto, houve contaminação pré e pós-pasteurização, demonstrando a necessidade de melhores práticas higiênicas, tanto na obtenção da matéria-prima, quanto na manipulação durante as diversas etapas de fabricação dos queijos.

Detection by multiplex PCR of Staphylococcus aureus , S. intermedius and S. hyicus in artificially contaminated milk / Detecção de Staphylococcus aureus , S. intermedius e S. hyicus em leite contaminado artificialmente por multiplex PCR

ABSTRACT: This research aimed to detect coagulase-positive Staphylococcus (CPS) directly in samples of artificially contaminated milk, using multiplex PCR (mPCR). Standard and isolated bacterial strains of S. aureus, S. epidermidis, S. hyicus, S. intermedius, Listeria monocytogenes and Escherichia coli species were used, evaluating the specificity and detection limit of mPCR, for artificially contaminated UHT milk. Primers specific for the nuc gene (NUC1-NUC2 were used for S. aureus, NUC3-NUC4 for S. hyicus and NUC5-NUC6 for S. intermedius). It was possible to detect the three target species by mPCR, directly from bovine whole milk, with adequate specificity and acceptable detention limit for identification of coagulase-positive Staphylococcus (CPS) in foods. The specificity was determined by the amplification of species-specific fragments, and the detection limit was assessed by the detection thresholds obtained for the three species (103 CFU mL-1). From these results, the mPCR described, with the proposed set of primers, has the potential for use in precise identification and differentiation between CPSs in milk samples.

RESUMO: Esta pesquisa teve como objetivo detectar diretamente em amostras de leite contaminado artificialmente Staphylococcus coagulase positiva (ECP) por multiplex PCR (mPCR). Cepas padrão e isolados de S. aureus, S. epidermidis, S. hyicus, S. intermedius, Listeria monocytogenes e Escherichia coli foram utilizados no estudo. Foram utilizados primers específicos para o gene nuc (NUC1-NUC2 para o S. aureus, NUC3-NUC4 para o S. hyicus e NUC5-NUC6 para o S. intermedius ). Foi possível detectar as três espécies-alvo por mPCR, formar diretamente nas amostras de leite integral bovino, com especificidade adequada e limite de detecção aceitável para identificação de espécies de Staphylococcus coagulase positiva (ECP) em alimentos. A especificidade foi determinada por meio da amplificação de fragmentos específicos das espécies e o limite de detecção foi avaliado pelos limiares de detecção obtidos para as três espécies (103 UFC mL-1 para as espécies presentes nas amostras de leite contaminadas artificialmente).

Insights into the genome of the xanthan-producing phytopathogen Xanthomonas arboricola pv. pruni 109 by comparative genomic hybridization.

The phytopathogenic bacterium Xanthomonas arboricola pv. pruni is the causal agent of Prunus Bacterial Spot disease that infects cultivated Prunus species and their hybrids. Furthermore, X. arboricola pv. pruni (Xap) plays a role in biotechnology since it produces xanthan gum, an important biopolymer used mainly in the food, oil, and cosmetics industry. To gain first insights into the genome composition of this pathovar, genomic DNA of X. arboricola pv. pruni strains was compared to the genomes of reference strains X. campestris pv. campestris B100 (Xcc B100) and X. campestris pv. vesicatoria 85-10 (Xcv 85-10) applying microarray-based comparative genomic hybridizations (CGH). The results implied that X. arboricola pv. pruni 109 lacks 6.67% and 5.21% of the genes present in the reference strains Xcc B100 and Xcv 85-10, respectively. Most of the missing genes were found to be organized in clusters and do not belong to the core genome of the two reference strains. Often they encode mobile genetic elements. Furthermore, the absence of gene clusters coding for the lipopolysaccharide (LPS) O-antigens of Xcc B100 and Xcv 85-10 indicates that the structure of the O-antigen of X. arboricola pv. pruni 109 differs from that of Xcc B100 and Xcv 85-10.

Genotypic profile of Listeria monocytogenes isolated in refrigerated chickens in southern Rio Grande do Sul, Brazil / Perfil genotípico de Listeria monocytogenes isolada em frango refrigerados comercializados na região sul do Rio Grande do Sul, Brasil

ABSTRACT: Listeria monocytogenes is of notable concern to the food industry, due to its ubiquitous nature and ability to grow in adverse conditions. This study aimed to determine the genotypic profile of L. monocytogenes strains isolated from refrigerated chickens marketed in the southern part of Rio Grande do Sul, Brazil. The strains of L. monocytogenes isolated were characterized by serotyping and Pulsed Field Gel Electrophoresis (PFGE). Three different serotypes (1/2a, 1/2b and 4e) were evaluated by PFGE, and the macrorestriction patterns utilizing enzymes AscI and ApaI, revealed five different pulsotypes. The presence of such varied genotypic profiles demonstrates the prevalence of L. monocytogenes contamination of chicken processing environments, which combined with ineffective cleaning procedures, allowing the survival, adaptation and proliferation of these pathogens, not only in the processing environment, but also in local grocery stores.

RESUMO: Listeria monocytogenes é uma notável preocupação para a indústria de alimentos, devido à sua natureza ubíqua e a capacidade de se multiplicar em condições adversas. Este estudo objetivou determinar o perfil genotípico de L. monocytogenes isolada a partir de frangos refrigerados comercializados na região sul do Rio Grande do Sul, Brasil. As cepas de L. monocytogenes foram selecionadas e caracterizadas por sorotipagem e Eletroforese em Gel de Campo Pulsado (PFGE). Três sorotipos diferentes (1/2a, 1/2b e 4e) foram avaliados por PFGE, e a combinação dos padrões de macrorestrição utilizando as enzimas AscI e ApaI revelou cinco diferentes pulsotipos. A presença de diferentes perfis genotípicos demonstra a importância da contaminação no ambiente de processamento de frangos, o qual, juntamente com procedimentos de limpeza ineficazes, permitem a sobrevivência, adaptação e proliferação desses patógenos, não somente no ambiente de processamento, mas também no local de comercialização destes produtos.

Microbiological and physicochemical characterization of the raw milk and the colonial type cheese from the Northwestern Frontier region of Rio Grande do Sul, Brazil / Caracterização microbiológica e físico-química de leite cru e queijo colonial da região Fronteira Noroeste do Rio Grande do Sul, Brasil

This study aimed at evaluating the microbiological and physicochemical characteristics of raw milk and colonial type cheese from Northwestern Frontier region of Rio Grande do Sul, Brazil. For this purpose, the samples were collected in January and July. Microbiological analyses (aerobic mesophilic bacteria, total/thermotolerant coliform, coagulase-positive Staphylococcus, Salmonella spp. and Listeria monocytogenes, lactic acid bacteria) and physicochemical assays (pH, acidity, total solids, protein, fat, aw, moisture, NaCl) were performed. The milk and cheese samples showed low microbiological quality because high counting of aerobic mesophilic bacteria, total/thermotolerant coliform and coagulase-positive Staphylococcus were detected. High counting of lactic acid bacteria was observed. However, neither Salmonella spp. nor Listeria monocytogenes was found. The standard deviations above one (1.0) in the fat, protein, moisture and salt contents indicated that no standard procedure was followed for producing the local cheese. The sample collection period caused differences in the microbiota, total solids of milk and cheese moisture contents, aw and salt. The maturation period did not significantly influenced on the microbial counts, but it provided an increase in protein contents and a decrease in aw value in cheese samples collected in July.

Este estudo avaliou as características microbiológicas e físico-químicas de leite cru e queijo colonial da região Fronteira Noroeste do Rio Grande do Sul, Brasil. Para isto, foram feitas coletas de amostras em janeiro e julho. Análises microbiológicas (bactérias aeróbias mesófilas, coliformes totais e termotolerantes, Staphylococcus coagulase positiva, Salmonella spp. e Listeria monocytogenes, bactérias ácido lácticas) e físico-químicas(pH, acidez, sólidos totais, proteína, lipídeos, Aa, umidade, NaCl) foram realizadas. As amostras de leitee de queijo indicaram baixa qualidade microbiológica, pois houve detecção de altos níveis de bactériasmesófilas, coliformes totais e termotolerantes e Staphylococcus coagulase positiva. Altas contagens de bactérias ácido láticas foram observadas. Entretanto, não foi detectada a presença de Salmonella spp. eListeria monocytogenes. O desvio padrão acima de um (1,0) nos conteúdos de lipídeos, proteínas, umidade e sal indicou que não houve seguimento do procedimento padrão estabelecido na produção local de queijos. O período de coleta de amostras resultou em diferenças nas análises de microbiota, sólidos totais do leitee dos queijos, o teor de umidade, Aa e sal. O período de maturação não causou significativa influênciasobre as contagens microbianas, mas promoveu aumento no conteúdo de proteína e diminuição na Aa dos queijos coletados em julho.

EStafilococos coagulase positiva em queijos minas frescal e minas padrão comercializadosem Pelotas, Rio Grande do Sul / Coagulase positive staphylococcus In minas frescal and minas padrão cheese commercialized in Pelotas, Rio Grande do Sul (Brazil)

O objetivo deste estudo foi avaliar a presença de Estafi lococos coagulase positiva (ECP) em amostras de queijos industrializados Minas Frescal e Minas Padrão, comercializados na cidade dePelotas, Rio Grande do Sul. Foram avaliadas vinte e oito (28) amostras de queijo minas frescal e quarenta e quatro (44) de queijo minas padrão, coletadas no comércio local. Quatro amostrasapresentaram contaminação acima do padrão estabelecido pela RDC 12, de 12 de janeiro de 2001 (ANVISA), sendo três de queijo minas frescal e uma de queijo minas padrão. A presença de ECPé preocupante uma vez que esses micro-organismos podem se multiplicar, produzir e secretar toxinas em níveis sufi cientes para causar intoxicação alimentar estafi locócica, enfermidade transmitida por alimentos de grande importância em saúde pública.

Detecção de genes do cluster egc em Staphylococcus aureus isolados de alimentos de origem animal / Detection of genes of egc cluster in Staphylococcus aureus isolated from foods of animal origin

Os objetivos deste estudo foram detectar, por PCR, genes codificadores de enterotoxinas estafilocócicas, pertencentes ao cluster egc (genes seg, sei, selm, seln e selo) em Staphylococcus aureus isolados em diferentes alimentos de origem animal, e relacionar sua presença com a fonte de isolamento. Quarenta e uma cepas de S. aureus de diferentes origens (carne de frango, leite cru, embutidos cárneos e queijo) foram avaliadas por PCR, por meio da amplificação de um fragmento de 3375pb (denominado egc parcial), que foi utilizado como marcador da presença do cluster, e fragmentos de cada um dos genes pertencentes ao cluster egc. Há presença de genes do cluster egc em isolados de S. aureus isoladas em alimentos de origem animal; entretanto, diferentes genótipos puderam ser observados em função da fonte de isolamento. A ocorrência de S. aureus isolados em carne de frango que possuíam todos os genes do cluster foi elevada; no entanto, nos isolados oriundos dos demais alimentos, essa ocorrência foi reduzida.

The aim of this study was to detect, through PCR usage, the genes which encodes staphylococcal enterotoxins and which belongs to egc cluster (seg, sei, selm, seln and selo) in S. aureus isolated from different foods of animal origin and correlate their presence with the strain origin. Forty-one strains of S. aureus from different sources (chicken meat, raw milk, sausage meat and cheese) were evaluated through PCR by amplifying a fragment of 3375bp (called partial egc), which was used as a marker for the presence of cluster, and fragments of individual genes belonging to egc cluster. There is presence of the egc cluster in strains of S. aureus isolated from foods of animal origin, however, different genotypes could be observed depending on the isolation source. The occurrence of strains isolated from chicken meat that had all the genes of the cluster was high; however, in the strains isolated from the other foods, such occurrence has been reduced.

AFLP analysis of Xanthomonas axonopodis and X arboricola strains used in xanthan production studies reveal high levels of polymorphism

Amplified fragment length polymorphism (AFLP) was used to analyze the genetic diversity of 14 strains of Xanthomonas arboricola pv. pruni and seven strains of X. axonopodis pv. phaseoli, which are used in xanthan production studies. Relationships identified by the AFLP profiles were assessed for xanthan production capacity, geographical location and host plant. Strains were isolated from 10 different geographic regions in South and Southeast States in Brazil. Data were analyzed for genetic similarity using the Dice coefficient and subjected to UPGMA cluster analysis. A total of 128 AFLP fragments were generated from four primer combinations: EcoRI+C/MseI+0, EcoRI+A/MseI+0, EcoRI+G/MseI+T and EcoRI+G/MseI+A. Of these, 96.1 percent were polymorphic. X. axonopodis pv. phaseoli (S D = 0.27) was shown to be more polymorphic than X. arboricola pv. pruni (S D = 0.58). All 14 pathovar pruni strains were included in a single main group (S D = 0.58), while the pathovar phaseoli strains were divided into three separate groups, with one group containing five strains (S D = 0.38) and two isolated groups (S D = 0.31 and 0.27) composed of only one strain each. Species were distinguished by three and eight specific AFLP markers present in the pathovar phaseoli and the pathovar pruni, respectively. For the unique strain without xanthan production capacity (X. axonopodis pv. phaseoli str. 48), nine specific AFLP bands were found. There was no evidence that geographic area or host plant influenced genetic heterogeneity. Correlations between AFLP patterns and xanthan production capacity were found in some strains, but were not consistent enough to establish a relationship.

Characterization of Listeria monocytogenes isolated from a fresh mixed sausage processing line in Pelotas-RS by PFGE / Caracterização de Listeria monocytogenes isoladas em uma linha de processamento de lingüiça mista frescal em Pelotas-RS por PFGE

Listeria monocytogenes is a bacterium capable to adhere to the surfaces of equipment and utensils and subsequently form biofilms. It can to persist in the food processing environmental for extended periods of time being able to contaminate the final product. The aim of this study was to trace the contamination route of L. monocytogenes on a fresh mixed sausage processing line, from raw material to the final product. The isolates obtained were characterized by serotyping and molecular typing by pulsed-field gel electrophoresis (PFGE) using the restriction enzymes ApaI and AscI. L. monocytogenes was detected in 25 percent of the samples. The samples of raw material were not contaminated, however, the microorganism was detected in 21 percent of the environmental samples (food contact and non-food contact), 20.8 percent of the equipments, 20 percent of the food worker's hands, 40 percent of the mass ready to packaging and in all the final products samples, demonstrating that the contamination of final product occurred during the processing and the importance of cross contamination. PFGE yielded 22 pulsotypes wich formed 7 clusters, and serotyping yielded 3 serotypes and 1 serogroup, however, the presence of serotypes 4b and 1/2b in the final product is of great concern for public health. The tracing of contamination showed that some strains are adapted and persisted in the processing environment in this industry.

Listeria monocytogenes é uma bactéria com capacidade de formar biofilmes e de colonizar superfícies de equipamentos e utensílios. Esse microrganismo pode persistir por longos períodos em plantas de processamento de alimentos, sendo capaz de contaminar o produto final. O objetivo deste estudo foi traçar a rota de contaminação de L. monocytogenes em uma linha de processamento de lingüiça mista frescal, desde a matéria-prima até o produto final. Os isolados obtidos foram caracterizados por sorotipagem e por tipagem molecular, através de Pulsed Field Gel Electrophoresis (PFGE), usando as enzimas de restrição ApaI and AscI. L. monocytogenes foi detectada em 25 por cento das amostras. Nenhuma amostra da matéria-prima apresentava o patógeno, entretanto, o microrganismo foi detectado em 21 por cento das amostras ambientais (com e sem contato com o alimento), 20,8 por cento dos equipamentos, 20 por cento das amostras de mãos de manipuladores, 40 por cento da massa pronta para o embutimento, e em todas as amostras do produto final. Isso demonstra que a contaminação do produto final ocorreu durante o processamento, e a importância da contaminação cruzada. PFGE produziu 22 pulsotipos e a sorotipagem produziu 3 sorotipos e 1 sorogrupo, entretanto, a presença dos sorotipos 4b e 1/2b no produto final é de grande importância para a saúde pública. Os perfis de macrorestrição mostraram que algumas cepas são adaptadas e persistiram no ambiente de processamento dessa indústria.

Ocorrência e perfil de sensibilidade a antibióticos de Salmonella spp. isolada em cortes de carne suína comercializados em feiras-livres de Pelotas-RS / Ocurrence and antibiotic sensibility profile of Salmonella spp. isaolated from pork meat cuts commercialized in the open markets in Pelotas, RS (Brazil)

O objetivo deste estudo foi avaliar a ocorrência de Salmonella spp. em 15 amsotras de cortes de carne suína (chuleta, pernil, copa e costela), comercializados em feiras-livres na cidade de Pelotas-RS, verificar os sorovares prevalentes e testar os isolados frente a diversos antibióticos de importância em clínica médica (ácido nalidíxico, ampicilina, aztreonam, canamicina, carbenicilina, cefalotina, cefoxitina, ceftriaxona, ciprofloxacina, cloranfenicol, gentamicina, sulfonamida, tetraciclina e trimetoprina). Doze amostras (80 por cento) apresentaram contaminação por Salmonella enterica, sorovares infantis, Derby, Panama e Typhimurium. Todos os isolados foram sensíveis à trimetoprina, aztreonam, ciprofloxacina, ceftriaxona e cefoxitina. Quanto aos demais antibióticos, o padrão de sensibilidade variou conforme o sorovar. Além disso, 39,1 por cento dos isolados apresentaram-e multirresistentes.