Characterization of diarrheagenic Escherichia coli from different cattle production systems in Brazil.

Diarrheagenic E. coli (DEC) can cause severe diarrhea and is a public health concern worldwide. Cattle are an important reservoir for this group of pathogens, and once introduced into the abattoir environment, these microorganisms can contaminate consumer products. This study aimed to characterize the distribution of DEC [Shiga toxin-producing E. coli (STEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), and enteroaggregative E. coli (EAEC)] from extensive and intensive cattle production systems in Brazil. Samples (n = 919) were collected from animal feces (n = 200), carcasses (n = 600), meat cuts (n = 90), employee feces (n = 9), and slaughterhouse water (n = 20). Virulence genes were detected by PCR in 10% of animal samples (94/919), with STEC (n = 81) as the higher prevalence, followed by EIEC (n = 8), and lastly EPEC (n = 5). Animals raised in an extensive system had a higher prevalence of STEC (average 48%, sd = 2.04) when compared to animals raised in an intensive system (23%, sd = 1.95) (Chi-square test, P < 0.001). From these animals, most STEC isolates only harbored stx2 (58%), and 7% were STEC LEE-positive isolates that were further identified as O157:H7. This study provides further evidence that cattle are potential sources of DEC, especially STEC, and that potentially pathogenic E. coli isolates are widely distributed in feces and carcasses during the slaughter process.

Characterization of microbial ecology, Listeria monocytogenes, and Salmonella sp. on equipment and utensil surfaces in Brazilian poultry, pork, and dairy industries.

This study aimed to evaluate the level of counting by indicator microorganisms, identify the microbial ecology, detect Listeria monocytogenes and Salmonella sp., and determine the presence of virulence genes and biofilm formation. A total of 480 samples were collected from the surfaces of the equipment and utensils using sterile swabs for the detection of L. monocytogenes and Salmonella sp. and counting mesophilic aerobes, Enterobacteriaceae, Escherichia coli, and Pseudomonas sp. The microbial ecology was evaluated by sequencing the 16S rRNA gene. Genes for virulence and biofilm formation were analyzed and adhesion capacity was evaluated for L. monocytogenes and Salmonella sp. The mesophilic aerobe count was the highest in the dairy processing facility, followed by the pork and poultry slaughterhouses. L. monocytogenes was detected in all facilities, with the highest detection in the pork slaughterhouse, followed by the poultry and dairy facilities. Salmonella sp. was only detected in the dairy. Isolates of L. monocytogenes and Salmonella sp. showed poor adhesion to polystyrene surfaces, virulence genes, and biofilm formation. The frequent contaminants in the slaughterhouses were Pseudomonas, Acinetobacter, and Aeromonas in poultry, Acinetobacter, Pseudomonas, and Brevundimonas in pork, and Pseudomonas, Kocuria, and Staphylococcus in dairy. Our results provide useful information to understand the microbiological risks associated with contamination.

Food handling in the domestic environment: an online questionnaire study with respondents from 24 of 26 Brazilian states / Manipulação de alimentos no ambiente doméstico: um estudo realizado com emprego de questionário online respondido por habitantes de 24 dos 26 estados brasileiros

Using an online questionnaire, this study evaluated the profile of a Brazilian population's food handling practices in the home environment. The questionnaire, containing questions about domestic behavior in terms of hygiene and food handling, was built and available through social media sites. Information about the participants' profiles, their food pre-preparation, food preparation, and food post-preparation practices, and the occurrence of foodborne diseases (FBDs) was included in the questionnaire. A total of 701 responses were obtained. The interviewees included 78.31% female participants and 21.68% male participants, with an average age of 31.2 years. Nearly all (94.3%) had a complete or incomplete higher education. In the pre-preparation stage, the participants evaluated the shelf life (97.28%) and storage temperature (44.79%) of the products while purchasing them. Regarding food handling practices, only a few participants washed the food packages before storing them (31.95%) or removed hand jewelry or other adornments when washing food (61.48%). Most participants washed their hands (91.58%) and washed vegetables (99.28%). But a group of interviewees reported washing raw meat (27.81%) before preparing it. Cutting surfaces such as plastic (50.36%) and glass (49.36%) tops were the most prevalent in the study. Most respondents did not know how long they had been using their cutting boards (67.62%) and mentioned using the same surface to handle both raw and ready-to-eat products (84.17%). As for the preparation, most interviewees declared they did not check the food temperature during preparation (86.31%), ignoring the ideal cooking temperature (88.26%). Regarding the occurrence of FBDs, 79.17% of the interviewees reported having suspicious clinical signs associated with contaminated foods and 65.59% did not seek medical help. Thus, the participants demonstrated ignorance about adequate practices for food safety in the home environment, highlighting the need to conduct health education programs within the Brazilian population.(AU)

Este estudo teve como objetivo avaliar o perfil das práticas de manipulação de alimentos no ambiente domiciliar no Brasil utilizando um questionário online. Um questionário contendo perguntas sobre comportamento doméstico em nível de higiene e manipulação de alimentos foi construído e disponibilizado por redes sociais. O questionário continha informações sobre o perfil dos participantes, suas práticas de pré-preparo, preparo e pós-preparo de alimentos e a ocorrência de doenças transmitidas por alimentos (DTA). Obteve-se 701 respostas, os entrevistados foram 78,31% do sexo feminino e 21,68% do sexo masculino, com média de idade de 31,2 anos. A maioria (94,3%) possuia ensino superior completo ou incompleto. Na etapa de pré-preparo, os participantes avaliam o prazo de validade (97,28%) e a temperatura de armazenamento (44,79%) dos produtos no momento da compra. Em relação às práticas de manipulação dos alimentos, apenas alguns participantes lavavam as embalagens dos alimentos antes de armazená-los (31,95%) ou retiravam adornos ao lavar os alimentos (61,48%). A maioria dos participantes lavam as mãos (91,58%) e os vegetais (99,28%); entretanto, um grupo de entrevistados relatou lavar carne crua (27,81%) antes de prepará-la. Superfícies de corte como tábuas de plástico (50,36%) e de vidro (49,36%) foram os mais prevalentes no estudo. A maioria dos entrevistados não sabe há quanto tempo usa as tábuas de corte (67,62%) e utilizam a mesma superfície para manusear produtos crus e prontos para o consumo (84,17%). Quanto ao preparo, a maioria dos entrevistados declarou não verificar a temperatura dos alimentos durante o preparo (86,31%), ignorando a temperatura ideal de cozimento (88,26%). Em relação à ocorrência de DVA, 79,17% dos entrevistados relataram que já apresentaram sinais clínicos suspeitos associados a alimentos contaminados e 65,59% não procuraram atendimento médico. Nesse sentido, os participantes demonstraram desconhecimento sobre as práticas adequadas para a segurança dos alimentos no ambiente domiciliar, evidenciando a necessidade de realização de programas de educação em saúde com a população brasileira.(AU)

Pathogenic potential and antibiotic resistance of Yersinia enterocolitica, a foodborne pathogen limited to swine tonsils in a pork production chain from Southern Brazil.

In this study, we aimed to characterize the distribution of Yersinia enterocolitica in a pork production chain in Brazil, as well as the virulence profile and antibiotic resistance of the obtained isolates. Samples from 10 pig lots obtained from finishing farms (water, feed, and barn floors, n = 30), slaughterhouse (lairage floors, carcasses at four processing steps, tonsils, and mesenteric lymph nodes, n = 610), and processing (end cuts, processing environment, n = 160) were obtained in Paraná state, Brazil, and subjected to Y. enterocolitica detection by ISO 10,273. The obtained isolates were identified based on biochemical and molecular features (16 s rRNA, inv, bioserotyping) and subjected to PCR assays to detect virulence (ail, ystA, ystB, virF, myfA, fepA, fepD, fes, tccC, ymoA, hreP, and sat) and multidrug resistance-related genes (emrD, yfhD, and marC). Also, isolates were subjected to disk diffusion test to characterize their resistance against 17 antibiotics from 11 classes and to pulsed field gel electrophoresis (PFGE) after XbaI macro-restriction. Y. enterocolitica was detected in a single sample (tonsil), and the obtained three isolates were characterized as serotype O:3, harboring ail, ystA, virF, myfA, tccC, ymoA, hreP, emrD, yfhD, and marC, and resistant to all tested antibiotics. The three isolates presented identical macro-restriction profiles by PFGE, also identical to isolates obtained from Minas Gerais, other Brazilian state; one selected isolate was identified as biotype 4. Despite the low occurrence of Y. enterocolitica in the studied pork production, the virulence potential and the antibiotic resistance profiles of the isolates demonstrated their pathogenic potential, and the macro-restriction profiles indicate strains descending from a common subtype in the pork production chain of two Brazilian States.

Phenotypic and genotypic characterization of non-typhoidal Salmonella isolated from a Brazilian pork production chain.

Pork products are important sources of foodborne non-typhoidal Salmonella in Brazil where antibiotics are commonly used throughout the pork production process and this has the potential to selectively favor antibiotic-resistant strains. We characterized the genotypic and phenotypic diversity of S. enterica isolates (n = 41) that were isolated in Brazil. Isolates were collected from ten swine farms and one slaughterhouse. Whole-genome sequencing and in silico serotyping demonstrated that the S. enterica serovar Typhimurium was the most common serotype (n = 17), but eight additional servoars were identified. Isolates presented high similarity based on comparison of DNA sequences (minimum of 89.6%), and sequence variation grouped according to serotype. Eight multilocus sequence types were identified with ST19 being most common (n = 21). Several plasmids replicons were detected, with Col (RNAI) the most abundant (n = 30), followed by IncR (n = 22), IncI1 (n = 10) and IncA/C2 (n = 10). Minimum inhibitory concentration assays showed that the principle resistance phenotypes were for streptomycin (90.2%), tetracycline (87.8%), ampicillin (80.5%), chloramphenicol (70.7%) and ciprofloxacin (51.2%). Only two isolates were resistant to third-generation cephalosporins and no isolates were resistant to two tested carbapenems. Twenty-six unique antimicrobial-resistance genes were identified with blaTEM-1A and blaTEM-1B likely responsible for most beta-lactam resistance and floR responsible for most chloramphenicol resistance. Six strains were positive for mcr-1. At the time of collection, the sampled farms were adding ciprofloxacin to feed and this may have contributed to the high prevalence of resistance to this antibiotic. The high number of multidrug resistant Salmonella and the presence of multiple resistant genes and plasmids emphasize the diversity of Salmonella in the studied pork chain, specially from serotype Typhimurium.

Comparison of Meat Juice Serology and Bacteriology for Surveillance of Salmonella in the Brazilian Pork Production Chain.

This study assessed an enzyme-linked immunosorbent assay (ELISA) based assay to detect Salmonella in swine as a potential tool to predict the presence of Salmonella in swine carcasses. The following samples were collected from 10 swine batches: blood (n = 100); environment (barn floor, n = 10, and lairage floor, n = 10); meat juice (n = 100, obtained after defrosting of diaphragm); tonsils (n = 100); mesenteric lymph nodes (MLNs) (n = 100); and carcasses after bleeding (n = 100), after singeing (n = 100), after evisceration (n = 100), and after final rinsing (n = 100). Blood and meat juice were subjected to ELISA to detect antibodies against Salmonella, and other samples were subjected to Salmonella detection by ISO 6579. Salmonella was detected in 3 samples from barn floors, 7 lairage floors, 45 tonsils, 43 MLNs and in 3 carcasses. Based on ELISA, Salmonella positive samples were: 86 and 46 blood serum (20% and 40% cut-offs) and 68 and 46 meat juice (20% and 40% cut-offs). Optical density readings from blood serum and meat juice presented a high and significant correlation (r = 0.93, p < 0.001), and a substantial agreement for Salmonella detection (K = 0.69, ELISA 40% cut-off). The agreement between ELISA and microbiological analysis for Salmonella detection in pig carcasses were absent or poor, with the exception of results obtained by ELISA 40% cut-off from blood serum and meat juice with MLNs (K = 0.49 and 0.50, respectively) and tonsils (K = 0.29 and 0.30, respectively). Based on the obtained results, meat juice can be considered an alternative to blood serum as a matrix for ELISA for preliminary detection of Salmonella, allowing the identification of potential sources of contamination during slaughtering.

Influence of time and storage temperature on raw milk deteriorating microbiota / Influência do tempo e temperatura de estocagem sobre a microbiota deteriorante do leite cru

The quality of raw milk depends on initial microbial contamination and conditions of storage until industry processing. Considering the influence of time and storage temperature on raw milk microbiota, the objective of this work was to quantify and monitor the multiplication of these groups under different conditions. For this purpose, 41 samples of raw milk were collected immediately after milking, stored in the following storage conditions: 25 °C/2 h; 35 °C/2 h; 7 °C/24 h; 7 °C/48 h and 7 °C/60 h and analyses of aerobic mesophilic, psychrotrophic and proteolytic psychrotrophic microorganisms. The milk samples analyzed in the study had an initial mean count of mesophilic aerobes of 5.38 Log CFU/mL at Time Zero. The milk stored at 25 °C/2 h and 35 °C/2 h kept the mesophilic aerobic counts within the limits established by the legislation (5.48 Log CFU/mL), with an increase in counts of psychrotrophic and proteolytic microorganisms. When stored at 7 °C/24 h and 7 °C/48 h, the count of mesophiles exceeded the established parameters. A significant increase in the count of proteolytic psychrotrophs and psychrotrophs was also observed during storage at 7 °C from 24 h. The results of this study indicate that the temperature of 7 °C is not suitable for the milk conservation, since it was not able to control the microbial multiplication. Thus, the results contribute to the change in milk storage temperature proposed by the new Brazilian legislation.(AU)

A qualidade do leite cru depende da contaminação microbiana inicial e das condições de armazenamento até o processamento na indústria. Considerando a influência do tempo e da temperatura de armazenamento na microbiota do leite cru, o objetivo deste trabalho foi quantificar e monitorar a multiplicação desses grupos de microrganismos sob diferentes condições. Para tanto, foram coletadas 41 amostras de leite cru imediatamente após a ordenha, armazenadas nas seguintes condições de armazenamento: 25 °C/2 h; 35 °C/2 h; 7 °C/24 h; 7 °C/48 h e 7 °C/60 h para análise de microrganismos psicrotróficos, aeróbios mesófilos, psicrotróficos e proteolíticos. As amostras de leite analisadas no estudo apresentaram uma contagem média inicial de aeróbios mesófilos de 5.38 Log UFC/mL no Tempo Zero. O leite armazenado a 25 °C/2 h e 35 °C/2 h manteve as contagens aeróbias mesófilas dentro dos limites estabelecidos pela legislação (5,48 Log UFC/mL), com aumento nas contagens de microrganismos psicrotróficos e proteolíticos. Quando armazenado a 7 °C/24 h e 7 °C/48 h a contagem de mesófilos excedeu os parâmetros estabelecidos. Um aumento significativo na contagem de psicrotróficos e psicrotróficos proteolíticos também foi observado durante o armazenamento a 7 °C a partir das 24 h. Os resultados deste estudo indicam que a temperatura de 7 °C não é adequada para a conservação do leite, uma vez que não foi capaz de controlar a multiplicação microbiana. Assim, os resultados contribuem para a mudança na temperatura de armazenamento de leite proposta pela nova legislação brasileira.(AU)

Distribution, adhesion, virulence and antibiotic resistance of persistent Listeria monocytogenes in a pig slaughterhouse in Brazil.

Listeria monocytogenes is a relevant pathogen usually associated with meat and ready-to-eat products. This study aimed to assess the distribution, adhesion, virulence and antibiotic resistance of L. monocytogenes in a pork production chain. Environment, carcass and food samples (n = 894) were obtained from different steps of a pork production chain over a 6-month period (10 samplings), including from farms and the slaughterhouse (reception, slaughtering, processing, storage and end products). L. monocytogenes was detected in samples from the reception (lairage floor, 1/10), slaughtering (drains, 2/20) and cutting room stages (conveyor belts in the final packing stage - 11/20, knife - 1/40, and cutting boards - 1/20). Positive results for conveyor belts were recorded in seven consecutive samplings. L. monocytogenes isolates (n = 87) were characterized as belonging to serogroup IVb and presented positive PCR results for inlA, inlB, inlC, inlJ, hlyA, plcA, actA and iap. Isolates were selected according to the original samples (n = 31) and subjected to Pulsed Field Gel Electrophoresis (PFGE), demonstrating their high clonal identity (98.4-100%). According to PFGE results and their original samples, isolates were selected (n = 16) and subjected to phenotypic assay to assess their adhesion potential and tested for resistance against 15 antibiotics; all tested isolates presented weak adhesion potential and were resistant to ampicillin. The present study demonstrated the persistence of L. monocytogenes in the pork processing facility, indicating the potential risk for cross-contamination with a potential virulent and resistant clone.

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.

Quantification of psychrotrophic bacteria and molecular identification of Pseudomonas fluorescens in refrigerated raw milk / Quantificação de bactérias psicrotróficas e identificação molecular de Pseudomonas fluorescens em leite cru refrigerado

In this study, we investigated the contamination of refrigerated raw milk produced in the western region of Paraná, southern Brazil, with psychrotrophic microorganisms, aiming to assay the proteolytic activity of the isolates and to identify Pseudomonas fluorescens, the main proteolytic species associated with the spoilage of milk products. Raw milk samples from 50 dairy farms were submitted to the counting of psychrotrophic microorganisms, being the microbiota characterized by its mesophilic behavior and proteolytic capacity, besides molecular identification of P. fluorescens. Of the samples evaluated, 94% had psychrotrophic counts ranging from 3 to 7.1 log CFU mL-1, and 48.5% of these showed mesophilic behavior. Of the isolates, 48.0% had proteolytic activity in at least one evaluated temperature (21 and 30°C), and 39.3% had proteolytic activity in both temperatures. Among the 61 isolates submitted to molecular identification by polymerase chain reaction (PCR), 86.8% contained the expression of the 16S gene characteristic for P. fluorescens. In this study, we demonstrated that P. fluorescens is the most prevalent psychrotrophic bacteria species in raw refrigerated milk and their proteolytic ability poses high risks to the dairy industry.(AU)

No presente estudo, investigamos a contaminação do leite cru refrigerado produzido na região oeste do Paraná, sul do Brasil, com micro-organismos psicrotróficos, visando testar a atividade proteolítica dos isolados e identificar Pseudomonas fluorescens, a principal espécie proteolítica associada à deterioração de produtos lácteos. Amostras de leite cru de 50 fazendas leiteiras foram submetidas à contagem de micro-organismos psicrotróficos, caracterizando-se a microbiota por seu comportamento mesofílico e sua capacidade proteolítica, além de identificação molecular de P. fluorescens. Entre as amostras avaliadas, 94% apresentaram contagem psicrotrófica variando de 3 a 7,1 log UFC mL-1 e 48,5% destas apresentaram comportamento mesofílico. Entre os isolados, 48,0% apresentaram atividade proteolítica em pelo menos uma das temperaturas testadas (21 e 30°C) e 39,3% apresentaram atividade proteolítica em ambas as temperaturas. Entre os 61 isolados sub-metidos à identificação molecular por reação em cadeia da polimerase (PCR), 86,8% continham expressão do gene 16S característico de P. fluorescens. Neste estudo, demonstramos que P. fluorescens é a espécie de bactérias psicrotróficas mais prevalente em leite refrigerado cru e sua capacidade proteolítica promove elevados riscos de deterioração para a indústria de laticínios.(AU)

Occurrence of Salmonella sp. in poultry carcasses evaluated from the retail trade between 2007 and 2013 in Paraná state, Brazil / Ocorrência de Salmonella sp. em carcaças de frango avaliadas a partir do comércio varejista entre 2007 e 2013 no estado do Paraná, Brasil

Poultry meat is often involved as a vehicle for microorganisms that cause food-borne diseases. Salmonella sp. is a major pathogen involved in outbreaks around the world. Based on its importance, the objective of this study was to determine the occurrence of Salmonella sp. in frozen and chilled poultry carcasses slaughtered and marketed in the western region of Paraná state, Brazil. A total of 340 samples were collected between January 2007 and April 2013, with 66 (19.41%) carcasses positive for Salmonella sp. It can be concluded from the results obtained that the occurrence of Salmonella sp. in poultry carcasses remains high, even though it is a product marketed refrigerated or frozen, methods considered appropriate for food preservation. A lower occurrence result was expected due to the 14-year implementation of the national pathogen reduction program established by the Ministry of Agriculture, Livestock and Supply aimed at a gradual reduction in the occurrence of Salmonella sp. in these products through constant monitoring of carcasses immediately after slaughter.(AU)

A carne de aves é um alimento que frequentemente encontra-se envolvido como veículo de micro-organismos causadores de enfermidades, sendo Salmonella sp. um desses agentes comumente envolvidos nessas enfermidades em todo o mundo. Com isso, o objetivo deste estudo foi determinar a ocorrência de Salmonella sp. em carcaças de frango congeladas e refrigeradas, abatidas e comercializadas na região oeste do estado do Paraná. Um total de 340 amostras foram coletadas entre janeiro de 2007 e abril de 2013. Das amostras analisadas, 66 (19,41%) das carcaças foram positivas para Salmonella sp. De acordo com os resultados obtidos pode-se concluir que a ocorrência de Salmonella sp. em carcaças de frangos permanece alta, mesmo sendo produtos comercializados refrigerados ou congelados, métodos considerados adequados para conservação de alimentos. Um resultado de menor ocorrência era esperado devido à implementação de 14 anos do Programa Nacional de Redução de Patógenos estabelecido pelo Ministério da Agricultura, Pecuária e Abastecimento visando uma redução gradual na ocorrência de Salmonella sp. nestes produtos através de monitoramento constante de carcaças imediatamente após o abate.(AU)

Yersinia enterocolitica in a Brazilian pork production chain: Tracking of contamination routes, virulence and antimicrobial resistance.

This study aimed to track Yersinia enterocolitica contamination in a pork production chain in Minas Gerais, Brazil, and to characterize the virulence and antibiotic resistance of isolates. Samples were collected from four different steps of the pork production chain (pig farm, carcass, processing environment and end product; n = 870), and tested for the presence of Y. enterocolitica. The pathogen was detected in 8 samples (palatine tonsils = 5; mesenteric lymph nodes = 2; carcass after bleeding = 1), from which 16 isolates were obtained and identified as Y. enterocolitica bioserotype 4/O:3. XbaI macrorestriction allowed the clustering of isolates in 5 pulsetypes, and the identification of identical profiles of Y. enterocolitca isolated from different samples. All isolates were positive for the virulence related genes ail, virF, myfA, ystA, tccC, ymoA, hreP and sat, and negative for ystB, ystC, fepA, fepD and fes. Considering 17 antibiotics from 11 classes, only ciprofloxacin and kanamycin were effective against all isolates, and three multidrug resistance profiles were identified among them, with simultaneous resistance to 9 of 11 classes. All isolates presented positive results for emrD, yfhD and marC, related to multidrug resistance. The results of this study demonstrated the contamination routes of Y. enterocolitica within the assessed pork production chain, and highlighted the pathogenic potential and antibiotic resistance of this foodborne pathogen.

Characterization of Tifton 85 bermudagrass haylage with different layers of polyethylene film and storage time.

OBJECTIVE: The objective was to characterize the fermentative and microbiological profile of Tifton 85 bermudagrass haylage with different layers of polyethylene film and storage time. METHODS: The experimental design consisted of a randomized block design with four and six wrapping layers (100 and 150 microns in total. respectively) allocated in the main plots, through repeated measures analysis (30, 60, and 90 days of storage) with four replicates. RESULTS: The storage time and number of wrapping layers did not show changes in the population of Clostridium and lactic acid bacteria. A decrease was observed in the enterobacteria population with an increase in the storage period in the two wrapping layers studied. Upon opening of the haylage at 30 days, the population of Bacillus was lower in haylages made with six layers of wrapping (3.63 log colony forming units/g). No growth of Listeria sp. or Salmonella sp. was observed during the experimental period. The fungal genera with a greater occurrence were Penicillium sp. and Fusarium sp. The following mycotoxins were not detected: ochratoxin A, fumonisins, and zearalenone. Relative to the organic butyric, propionic, and acetic acids, the haylages presented a low concentration of lactic acid; this may have prevented a drop in the pH, which was high when the silos were opened (5.4). The levels of ammoniacal nitrogen and soluble carbohydrates presented no variation among the number of wrapping layers, with an overall average of 35.55 and 38.04 g/kg. CONCLUSION: Tifton 85 bermudagrass haylage wrapped with four and six layers presented adequate fermentation and microbiological characteristics in the evaluated periods.

Consumer profile and problems associated with uninspected raw milk consumption in western Paraná / Perfil dos consumidores e problemas associados ao consumo de leite cru informal no oeste do Paraná

The objectives of this study were to analyze consumer profile; to identify the main reasons for raw milk consumption; and to analyze in laboratory samples of uninspected raw milk from five towns in the western region of the state of Paraná, Brazil. The types of milk most frequently consumed were: 42.3% ultra-high temperature (UHT), 38.3% pasteurized milk, 17.6% uninspected raw milk, and 1.7% powered milk. The frequencies of households that preferred uninspected raw milk were, according to the town, 32.7% in Iporã, 29.2% in Marechal Cândido Rondon, 18.9% in Assis Chateaubriand, 17.6% in Palotina, and 10% in Toledo. Flavor was the main reason for uninspected raw milk consumption, and the purchase of this product was more frequent in households whose income was between one to four minimum wages. It was observed that the sales of uninspected milk are more financially advantageous to the producer than sales of inspected milk. All samples analyzed showed lack of compliance with at least one parameter, 60.9% for mesophilic counts, 56.6% for non-fat dry matter, 52.1% for freezing point, 43.5% for acidity, 23.9% for density, 23.9% for the casein macropeptide, 17.4% for fat content, 8.7% were reactors in the milk ring test, and 2.2% were reactors in microbial growth inhibitor test. Fraud by addition of water was observed in 20% of the samples. Uninspected raw milk analyzed in this study involved a low-quality product that is a financial hazard as it may be adulterated, and that poses risk to consumer health.(AU)

Os objetivos desta pesquisa foram verificar o perfil, identificar os principais motivos para o consumo e avaliar laboratorialmente as amostras de leite cru informal de cinco municípios do oeste do estado do Paraná, Brasil. Os tipos de leite mais frequentemente consumidos foram: 42,3% ultra-high temperature (UHT), 38,3% pasteurizado, 17,6% leite cru informal e 1,7% em pó. As frequências de consumidores que preferiram leite cru informal, por cidade, foram: 17,6% em Palotina, 29,2% em Marechal Cândido Rondon, 18,9% em Assis Chateaubriand, 17,6% em Palotina e 10% em Toledo. Sabor foi o principal motivo que influenciou o consumo de leite cru informal, e famílias com renda familiar de um a quatro salários mínimos foram as que mais consumiram esse tipo de leite. A comercialização informal de leite foi mais vantajosa financeiramente para o produtor em relação à comercialização formal. Todas as amostras analisadas apresentaram-se em desacordo em pelo menos um parâmetro, sendo 60,9% para mesófilos, 56,6% para estrato seco desengordurado, 52,1% para índice crioscópico, 43,5% para acidez, 23,9% para densidade, 23,9% para índice de caseíno macropeptídeo, 17,4% para teor de gordura, 8,7% reagente ao teste do anel em leite (TAL), e 2,2% reagente para presença de inibidor de crescimento bacteriano. A adulteração encontrada foi a adição de água em 20% das amostras analisadas. O leite cru informal avaliado nesta pesquisa envolveu a comercialização de um produto sem qualidade e que constitui um perigo financeiro, em razão do consumo de produtos adulterados, além do risco à saúde do consumidor.(AU)

Molecular tracking of Salmonella spp. in chicken meat chain: from slaughterhouse reception to end cuts.

Due to the importance of Salmonella spp. in poultry products, this study aimed to track its main contamination routes since slaughtering reception to processing of chicken end cuts. Samples from different steps of slaughtering and processing (n = 277) were collected from two chicken slaughterhouses (Sl1 and Sl2) located in Minas Gerais state, Brazil, and subjected to Salmonella spp. detection. The obtained isolates were subjected to serological identification and tested by PCR for specific Salmonella spp. genes (ompC and sifB). Also, Salmonella spp. isolates were subjected to XbaI macrorestriction and pulsed-field gel electrophoresis (PFGE). Sixty-eight samples were positive for Salmonella spp. and 172 isolates were obtained. Sl1 and Sl2 presented similar frequencies of Salmonella spp. positive samples during reception, slaughtering and processing (p > 0.05), except for higher frequencies in Sl1 for chicken carcasses after de-feathering and evisceration (p < 0.05). PFGE allowed the identification of cross contamination and persistence of Salmonella spp. strains in Sl1. The results highlighted the relevance of the initial steps of chicken slaughtering for Salmonella spp. contamination, and the pre-chilling of carcasses as an important controlling tool. In addition, the presence of Salmonella spp. in chicken end cuts samples represents a public health concern.

Multidrug resistance and ESBL-producing Salmonella spp. isolated from broiler processing plants

Abstract The aim of this study was to investigate the occurrence of multidrug-resistant, extended spectrum beta-lactamase (ESBL) producing Salmonella spp. isolated from conveyor belts of broiler cutting rooms in Brazilian broiler processing plants. Ninety-eight strains of Salmonella spp. were analyzed. Multidrug resistance was determined by the disk diffusion test and the susceptibility of the isolated bacteria was evaluated against 18 antimicrobials from seven different classes. The double disk diffusion test was used to evaluate ESBL production. Of the 98 strains tested, 84 were multidrug resistant. The highest rates of resistance were against nalidixic acid (95%), tetracycline (91%), and the beta-lactams: ampicillin and cefachlor (45%), followed by streptomycin and gentamicin with 19% and 15% of strain resistance, respectively. By contrast, 97% of the strains were sensitive to chloramphenicol. 45% of the strains were positive for the presence of ESBL activity. In this study, high rates of multidrug resistance and ESBL production were observed in Salmonella spp.

Multidrug resistance and ESBL-producing Salmonella spp. isolated from broiler processing plants.

The aim of this study was to investigate the occurrence of multidrug-resistant, extended spectrum beta-lactamase (ESBL) producing Salmonella spp. isolated from conveyor belts of broiler cutting rooms in Brazilian broiler processing plants. Ninety-eight strains of Salmonella spp. were analyzed. Multidrug resistance was determined by the disk diffusion test and the susceptibility of the isolated bacteria was evaluated against 18 antimicrobials from seven different classes. The double disk diffusion test was used to evaluate ESBL production. Of the 98 strains tested, 84 were multidrug resistant. The highest rates of resistance were against nalidixic acid (95%), tetracycline (91%), and the beta-lactams: ampicillin and cefachlor (45%), followed by streptomycin and gentamicin with 19% and 15% of strain resistance, respectively. By contrast, 97% of the strains were sensitive to chloramphenicol. 45% of the strains were positive for the presence of ESBL activity. In this study, high rates of multidrug resistance and ESBL production were observed in Salmonella spp.

Ripened Semihard Cheese Covered with Lard and Dehydrated Rosemary (Rosmarinus officinalis L.) Leaves: Processing, Characterization, and Quality Traits.

This study aimed to establish a manufacturing protocol and to characterize semihard cheese covered with lard and rosemary during ripening. After the manufacturing protocol was defined, the cheeses were produced with pasteurized and raw milk from Holstein cows, with and without (control) coating, and then ripened for 60 d. During this period the physicochemical properties, color, proteolysis, texture profile, and sensory acceptance were performed. The early-ripening cheeses differed from the others in terms of color and moisture content. Multivariate statistical analysis separated chesses in groups differentiated by the effects of heat treatment of milk and ripening period. The ripened cheeses obtained from raw milk were sensorially more preferred. The coating gave the final products higher moisture content and favored color and texture characteristics. Consumer testing showed that the cheese obtained from raw milk and coated with lard and rosemary was the most preferred (acceptance of 82%) due to the specific coating of rosemary (aroma and flavor). This product has potential to add value and to diversify the production of semihard cheeses.

Condições de comercialização de produtos de origem animal em feiras livres de municípios da região oeste do Paraná

O objetivo deste trabalho foi constatar a situação higienicossanitária de barracas de feiras livres localizadas em cinco municípios da região oeste do Paraná e a qualidade microbiológica e físico-química dos produtos nelas comercializados. Entre fevereiro e maio de 2013, foram realizadas avaliações, por meio do uso de checklist em 52 barracas onde foram adquiridas 71 amostras sendo estas submetidas à análise microbiológica e/ou físico-química. Foi constatado que em 47,9% das barracas o armazenamento dos alimentos era realizado de forma inadequada, que 86% dos produtos de origem animal estavam acondicionados em embalagens impróprias e que em 65% das mesmas, a condição higiênica dos manipuladores foi classificada como "Regular". Foram encontradas não conformidades em 36,6% das amostras submetidas às análises microbiológicas e/ou físico-químicas, sendo detectada a presença de Salmonella sp. em uma amostra de linguiça colonial. Sendo assim, é necessário que haja um controle sanitário na produção e comércio dos produtos nestas feiras por parte dos órgãos públicos responsáveis, garantindo assim, a segurança alimentar do consumidor.

The objective of this study was to determine the hygiene conditions of street market stands that sell products of animal origin in five cities in western Parana, Brazil, as well as the microbiological and physical-chemical quality of the products that are sold there. Between February and May, 2013, a total of 52 street market stands were analyzed based on a checklist. 71 samples were collected in these stands and submitted to microbiological and/or physical-chemical analyses. It was observed that in 47.9% of the stands, food products were inadequately stored; 86% of the products of animal origin were kept in improper containers; and in 65% of the stands, handler hygiene was classified as regular. 36.6% samples did not comply with microbiological and/or physical-chemical standards, with one sample of colonial sausage showing the presence of Salmonella sp. Therefore, public organisms control of hygiene in the production and commercialization of products in street markets is highly necessary to ensure food safety.

Behavior of Staphylococcus aureus and autochthone microbiota in fresh sausages added of sodium nitrite and stored under refrigeration / Comportamento de Staphylococcus aureus e microbiota autóctone frente à ação de nitrito de sódio em linguiças frescais estocadas sob refrigeração

Fresh sausages are cured meat products that may be contaminated with Staphylococcus aureus during the manufacturing procedure, which is frequently related with inadequate handling practices. The use of nitrite in meat products has proven efficacy against Clostridium botulinum, and studies indicate that bactericidal action against S. aureus depends on factors that are intrinsic and extrinsic to the product. The objective of the present study was to evaluate the effect of nitrite concentration, and pH on S. aureus and psychrotrophic autochthone microbiota in fresh sausages stored at different times and temperatures. Fresh sausage were produced at nitrite concentrations 50, 150 and 200ppm and contaminated with S. aureus. The sausages were storage at refrigeration (7 and 12°C) and the quantification of S. aureus and psychrotrophic microorganisms was carried out on days 0, 2, 4, 7, and 10. Results showed that nitrite concentrations and the temperatures used had minimal effect on the multiplication of S. aureus and psychrotrophic autochthone microbiota. Final counts depended only on the length of storage: at the end of 10 days, counts were statistically similar in the different groups, showing that temperature and nitrite concentrations used did not control microbial growth effectively. It is suggested that the product should be stored below 7°C or at freezing temperatures for greater microbiological stability.

A linguiça frescal é um embutido curado que, devido à manipulação durante as etapas de produção, pode se tornar contaminado com S. aureus, patógeno frequentemente relacionado com práticas inadequadas de higiene durante a produção de alimentos. A utilização de nitrito em embutidos tem sua eficácia comprovada contra C. botulinum e, para o S. aureus, estudos indicam que a ação bactericida depende de fatores extrínsecos e intrínsecos. O objetivo do presente estudo foi avaliar o efeito da concentração de nitrito e do pH frente à contaminação por S. aureus e microbiota autóctone em linguiças frescais estocadas em diferente tempos e temperaturas. Linguiças foram produzidas com concentrações de 50, 150 e 200ppm de nitrito e contaminadas com S. aureus. As linguiças foram armazenadas sob refrigeração (7 e 12°C) e a quantificação de S. aureus e psicrotróficos foi realizada nos dias 0, 2, 4, 7 e 10. Os resultados demonstraram que a influência das concentrações de nitrito e temperaturas utilizadas sobre a multiplicação de S. aureus e da microbiota autóctone foi mínima, sendo dependente apenas do período de estocagem. Entretanto, ao final de dez dias, as contagens foram estatisticamente iguais nos grupos analisados, mostrando que as condições de temperatura e concentrações de nitrito utilizadas não exerceram controle efetivo no desenvolvimento destes micro-organismos. Sugere-se que este produto seja armazenado sob temperaturas inferiores a 7°C ou sob congelamento para maior estabilidade microbiológica.