Bacterial communities in artisanal raw bovine milk cheeses from the southern region of Brazil / Comunidades bacterianas em queijos artesanais de leite bovino cru da região sul do Brasil

The production of artisanal cheeses made with raw bovine milk has grown in the southern region of Brazil. It is important to obtain information about the risks of this practice, especially concerning food safety. In this study, next-generation sequencing was used to identify and characterize the bacterial communities of artisanal raw milk cheeses. We analyzed one pool of five raw milk samples (control group M1) from different dairy farms and nine pools (M2-M10) of 45 artisanal raw milk cheeses.The characterization of the bacterial communities included 199 species distributed across 59 different genera dispersed among the samples. Among the genera observed, 11 were classified as beneficial to the aroma, flavour, colour, and texture of the cheese. Thirty-one genera were classified as harmful to these characteristics. Another 17 were classified as potential pathogens for animals and humans, including Aeromonas, Bacillus, Cronobacter, Salmonella, Staphylococcus, and bacteria of the coliform group, including E. coli and Klebsiella. There was a significant difference (P < 0.05) in the number of bacterial communities identified between the control group (M1) and the two pools of artisanal raw milk cheeses (M2 and M8). This study demonstrated that next-generation sequencing provides in-depth information on the composition of the microbiota in artisanal raw milk cheeses, characterizing bacterial communities, identifying the wide microbial diversity, and identifying microbial benefits and risks.

Devido ao aumento da produção de queijos artesanais com leite bovino cru na região sul do Brasil, é importante obter informações sobre os riscos desta prática, principalmente no que se refere à segurança do alimento. Neste estudo foi utilizada a técnica de Next Generation Sequencing (NGS) para identificar e caracterizar comunidades bacterianas de queijos artesanais de leite cru. Foram analisados um pool de cinco amostras de leite cru como grupo controle (M1) de diferentes propriedades leiteiras localizadas na região norte do estado do Rio Grande do Sul, e nove pools (M2-M10) de 45 queijos artesanais de leite cru. A caracterização das comunidades bacterianas incluiu 199 espécies distribuídas em 59 gêneros diferentes dispersos entre as amostras. Dentre os gêneros observados, 11 foram classificados como benéficos ao aroma, sabor, cor e textura do queijo, enquanto 31 gêneros foram classificados como prejudiciais a essas características. Outros 17 foram classificados como potenciais patogênicos para animais e humanos, incluindo Aeromonas, Bacillus, Cronobacter, Salmonella, Staphylococcus, bactérias do grupo coliforme como Escherichia coli e Klebsiella. Houve diferença significativa (P < 0,05) entre o número de comunidades bacterianas identificadas no grupo controle (M1) e dois pools de queijos artesanais de leite cru (M2 e M8). Este estudo demonstra que o NGS fornece informações detalhadas sobre a composição da microbiota em queijos artesanais de leite cru, caracterizando comunidades bacterianas, identificando a ampla diversidade microbiana, os benefícios e riscos microbianos.

Biofilm forming antimicrobial-multiresistant Staphylococcus aureus / Multirresistência antimicrobiana de Staphylococcus aureus formadores de biofilme

Staphylococcus aureus is one of the main agents isolated from bovine mastitis cases, characterized by lower cure rates compared to other pathogens causing this disease. This phenomenon is mainly explained by the multiresistance acquisition to antimicrobials and the ability of S. aureus to form biofilms on biotic and abiotic surfaces. In this work 15 samples of S. aureus isolated from the automated milking facility were analyzed regarding the resistance profile to antimicrobials, virulence factors (capsule production, hemolysin, and protease) and adhesion capacity under different temperatures (42±1°C, 36±1°C, 25±1°C, 9±1°C, and 3±1°C). All isolates showed methicillin-resistant (MRSA) characteristics and multidrug resistance profile to the antimicrobials tested (penicillin G, chloramphenicol, oxacillin, cephalexin, tetracycline, amoxicillin + clavulanic acid, sulfa + trimetropim, gentamicin, doxycycline, ceftiofur, neomycin, and vancomycin) with an IRMA index between 0.5 and 1.0. Five isolates were resistant to vancomycin (VRSA), two were resistant to all active principles, and the others to at least six of these drugs. Adhesion capacity and biofilm formation were found in 3 of the 5 evaluated temperatures, including the cooling conditions. Regarding the virulence factors, 86.7% of the isolates formed capsules, 60% revealed the presence of protease, 26.7% expressed the α-hemolysin factor, and 13.3% of them presented β-hemolysin. The fact that all isolates presented MRSA characteristics represents a potential risk to those exposed to this agent, and the formation of biofilm in liners even after the use of detergents and sanitization highlights the urgency of searching for alternatives for dispersion of the biofilm by S. aureusin the automated milking facility.

O Staphylococcus aureus é um dos principais agentes isolados de casos de mastite bovina, caracterizado por menores taxas de cura em comparação com outros patógenos desta enfermidade. Esse fenômeno é explicado principal-mente pela aquisição de resistência à antimicrobianos e a capacidade do S. aureus formar biofilmes em superficies bióti-cas e abióticas. Neste trabalho foram utilizadas 15 amostras de S. aureus isolados de ordenhadeira, analisados quanto ao perfil de resistência à antimicrobianos, fatores de virulência (produção de cápsula, hemolisina e protease) e capacidade de adesão sob diferentes temperaturas (42±1°C, 36±1°C, 25±1ºC, 9±1ºC e 3±1ºC). Todos os isolados apresentaram perfil de multirresistência aos antimicrobianos testados (penicilina G, cloranfenicol, oxacilina, cefalexina, tetraciclina, amoxicilina + ácido clavulônico, sulfa + trimetropim, gentamicina, doxiciclina, ceftiofur, neomicina e vancomicina) com índice IRMA entre 0,5 a 1,0. Duas cepas foram resistentes a todos os princípios ativos e as demais a pelo menos seis destes fármacos. Os isolados avaliados apresentaram característica de meticilina-resistentes (MRSA) e destes, 33,34% (5/15) foram resistentes à vancomicina (VRSA). Houve capacidade de adesão e formação de biofilmes em 3 das 5 tem-peraturas avaliadas, incluindo as temperaturas de refrigeração. Em relação aos fatores de virulência, 86,7% dos isolados formaram cápsula, 60% presença de protease, 26,7% expressaram o fator α-hemolisina e 13,3% β-hemolisina. O fato de todos isolados apresentarem característica MRSA representa um risco potencial aos expostos a esse agente. Já a for-mação de biofilmes em teteiras, mesmo após detergência e sanitização, destacam a urgência de alternativas de dispersão de biofilmes no ambiente de ordenha.

Lactic acid bacteria against Listeria monocytogenes

Listeria monocytogenes is a pathogenic bacterium that can contaminate food and cause public health problems due its ability to form biofilms and resistance to sanitizers, it is responsible for sanitary and economic losses in food producing establishments. The difficulties in controlling biofilms and increasing resistance to traditional antibacterial agents is motivating studies of alternative potential biological agents for the control of pathogenic biofilms, among which lactic acid bacteria (LABs) are included. The objective of this work was to evaluate the activity of LABs against Listeria monocytogenes biofilm formation on polystyrene plates, a surface commonly used in the food industry. Lyophilized commercial strains of Bifidobacterium animalis, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus salivaris and Lactobacillus acidophilus were used. The strain of Listeria monocytogenes (L4) was isolated from polystyrene mats from a poultry slaughterhouse cutting room and demonstrated the ability to attach to microplates and resistance to sanitizers (sodium hypochlorite and hydrogen peroxide) at all times, temperatures and tested surfaces. The antimicrobial activity of LABs was evaluated by the agar diffusion method. The LABs that presented action on Listeria monocytogenes were selected for the inhibition and/or removal of biofilms in microplates, and all experiments were carried out in triplicate. Only Bifidobacterium animalis and Lactobacillus plantarum demonstrated action against Listeria monocytogenes in the agar diffusion assays and were selected for inhibition and competition assays. Furthermore, competition of LABs against Listeria monocytogenes adhesion was evaluated. There was no significant difference between LABs and L. monocytogenes, alone or in combination, at temperatures of 30ºC and 37ºC in the Listeria monocytogenes inhibition assays on polystyrene surface. The lactic acid bacteria evaluated did not demonstrate inhibition of L. monocytogenes adhesin testes with optical density visualization, however, it was possible to identify a reduction in L. monocytogenes counts with the application of Bifidobacterium animals and Lactobacillus plantarum in the testes of competition against biofilm formation. In competition tests Bifidobacterium animalis and Lactobacillus plantarum have an injunction in Listeria monocytogenes, indicating that these lactic acid bacteria can retard Listeria biofilm formation on polystyrene surfaces and thus help control the pathogen in the food industry. A potential mechanism to control biofilm adhesion and formation of pathogens for nutrients and fixation on surfaces, multiplication factors and surfaces are a challenge in controlling biofilms of pathogenic microorganisms, alternative measures to traditional methods for inactivating pathogens and biofilm formers bacteria are necessary. In this sense, lactic acid bacteria generate high levels of bacteriocin and are effective in inhibiting the biofilm of pathogenic bacteria, however, our study did not reveal this. We verified that Bifidobacterium animalis and Lactobacillus plantarum have an inhibitory action on Listeria monocytogenes, indicating that these lactic acid bacteria can be used to delay the formation of biofilms by Listeria on polystyrene surfaces, helping to control this pathogen in food industry.(AU)

Biofilm forming antimicrobial-multiresistant Staphylococcus aureus / Multirresistência antimicrobiana de Staphylococcus aureus formadores de biofilme

Staphylococcus aureus is one of the main agents isolated from bovine mastitis cases, characterized by lower cure rates compared to other pathogens causing this disease. This phenomenon is mainly explained by the multiresistance acquisition to antimicrobials and the ability of S. aureus to form biofilms on biotic and abiotic surfaces. In this work 15 samples of S. aureus isolated from the automated milking facility were analyzed regarding the resistance profile to antimicrobials, virulence factors (capsule production, hemolysin, and protease) and adhesion capacity under different temperatures (42±1°C, 36±1°C, 25±1°C, 9±1°C, and 3±1°C). All isolates showed methicillin-resistant (MRSA) characteristics and multidrug resistance profile to the antimicrobials tested (penicillin G, chloramphenicol, oxacillin, cephalexin, tetracycline, amoxicillin + clavulanic acid, sulfa + trimetropim, gentamicin, doxycycline, ceftiofur, neomycin, and vancomycin) with an IRMA index between 0.5 and 1.0. Five isolates were resistant to vancomycin (VRSA), two were resistant to all active principles, and the others to at least six of these drugs. Adhesion capacity and biofilm formation were found in 3 of the 5 evaluated temperatures, including the cooling conditions. Regarding the virulence factors, 86.7% of the isolates formed capsules, 60% revealed the presence of protease, 26.7% expressed the α-hemolysin factor, and 13.3% of them presented β-hemolysin. The fact that all isolates presented MRSA characteristics represents a potential risk to those exposed to this agent, and the formation of biofilm in liners even after the use of detergents and sanitization highlights the urgency of searching for alternatives for dispersion of the biofilm by S. aureusin the automated milking facility.(AU)

O Staphylococcus aureus é um dos principais agentes isolados de casos de mastite bovina, caracterizado por menores taxas de cura em comparação com outros patógenos desta enfermidade. Esse fenômeno é explicado principal-mente pela aquisição de resistência à antimicrobianos e a capacidade do S. aureus formar biofilmes em superficies bióti-cas e abióticas. Neste trabalho foram utilizadas 15 amostras de S. aureus isolados de ordenhadeira, analisados quanto ao perfil de resistência à antimicrobianos, fatores de virulência (produção de cápsula, hemolisina e protease) e capacidade de adesão sob diferentes temperaturas (42±1°C, 36±1°C, 25±1ºC, 9±1ºC e 3±1ºC). Todos os isolados apresentaram perfil de multirresistência aos antimicrobianos testados (penicilina G, cloranfenicol, oxacilina, cefalexina, tetraciclina, amoxicilina + ácido clavulônico, sulfa + trimetropim, gentamicina, doxiciclina, ceftiofur, neomicina e vancomicina) com índice IRMA entre 0,5 a 1,0. Duas cepas foram resistentes a todos os princípios ativos e as demais a pelo menos seis destes fármacos. Os isolados avaliados apresentaram característica de meticilina-resistentes (MRSA) e destes, 33,34% (5/15) foram resistentes à vancomicina (VRSA). Houve capacidade de adesão e formação de biofilmes em 3 das 5 tem-peraturas avaliadas, incluindo as temperaturas de refrigeração. Em relação aos fatores de virulência, 86,7% dos isolados formaram cápsula, 60% presença de protease, 26,7% expressaram o fator α-hemolisina e 13,3% β-hemolisina. O fato de todos isolados apresentarem característica MRSA representa um risco potencial aos expostos a esse agente. Já a for-mação de biofilmes em teteiras, mesmo após detergência e sanitização, destacam a urgência de alternativas de dispersão de biofilmes no ambiente de ordenha.(AU)

Effects of different concentrations of sodium azide and chloramphenicol on the preservation of raw milk samples / Efeitos de diferentes concentrações de azida sódica e cloranfenicol na preservação de amostras de leite cru

The preservation of milk samples for microbiological analyses by the Brazilian Network of Milk Quality Control Laboratories requires the addition of preservatives to maintain the microbiota from the time of sample collection to the moment of analysis. The number of microorganisms can change as a result of the active ingredients and concentration of the preservative, as well as due to interactions between the preservatives, incubation time, and packaging temperature. The objective of this research was to evaluate the conservation potential of different concentrations of sodium azide and chloramphenicol on the analytical shelf life of milk samples. Two farms were selected, one with a low bacterial count and one with a high bacterial count. The milk was dispensed into sterile vials and tested after the addition of the usual concentrations of sodium azide and chloramphenicol, doubled concentrations, tripled concentrations, and as a control, without preservatives. The samples were incubated at 3 ± 1 °C, 6 ± 1 °C, and 9 ± 1 °C for 14 days and analyzed daily for their bacterial count by flow cytometry. The tripled preservative concentrations improved conservation, increasing the timespan of the analytical viability of the samples without altering the results.(AU)

A conservação das amostras de leite destinadas para análises microbiológicas pela Rede Brasileira de Laboratórios de Controle da Qualidade do Leite requer adição de conservantes para a preservação da microbiota existente desde o momento da coleta até as análises. O número de microrganismos pode apresentar alterações decorrentes do princípio ativo e concentração do conservante, e ainda entre as interações conservante, tempo de incubação e temperatura de acondicionamento. O objetivo deste trabalho foi avaliar o potencial de conservação de diferentes concentrações de azida sódica e cloranfenicol sobre a vida útil analítica de amostras de leite. Foram selecionadas duas fazendas, sendo uma com baixa contagem bacteriana e outra com alta contagem bacteriana. O leite foi fracionado em frascos estéreis e testado nas seguintes condições: pastilhas com a concentração usual de azida sódica e cloranfenicol, com dupla concentração, com tripla concentração e sem a adição do conservante. As amostras foram incubadas por quatorze dias a 3 ± 1 °C, 6 ± 1 °C e 9 ± 1 °C, e analisadas diariamente por citometria de fluxo para a determinação da contagem bacteriana. A tripla concentração do conservante demonstrou maior conservação, possibilitando o aumento da viabilidade analítica das amostras sem alteração nos resultados.(AU)

Pomegranate (Punica granatum L) peel lyophilized extract delays lipid oxidation in tuscan sausages / Estabilidade oxidativa de linguiça tipo Toscana com extrato liofilizado da casca de romã (Punica granatum L)

Pomegranate (Punica granatum L.) contains a high concentration of antioxidant and phenolic compounds. Pomegranate peel extracts at different concentrations were used as natural antioxidant to increased the lipid stability of fresh Tuscan sausages, comparing with sodium erythorbate (SE). Peels were subjected to ultrasound-assisted aqueous extraction and lyophilization. The extract was previously characterized throught phenol and flavonoids determination. The Tuscan Sausages were prepared, and color, pH, peroxides, and thiobarbituric acid reactive substances were assessed at 1, 15, and 30 days of storage, at a mean temperature of 5 ± 1 ºC and under a 12-hour light cycle/day. Sausages containing 0.05 and 0.1% of peel extract showed results similar to sodium erythorbate in lipid peroxidation. Sausages treated with pomegranate peel extracts showed an adequate global acceptability level in the sensory analysis. Pomegranate peel extracts are; therefore, a promising natural alternative for maintaining the lipid stability of Tuscan sausages, promoting the protection of the meat and in addition, being able to bring beneficial of the pomegranate to the human health.(AU)

A romã (Punica granatum L.) contém uma alta concentração de compostos antioxidantes e fenólicos. Extratos de casca de romã em diferentes concentrações foram utilizados como antioxidante natural para aumentar a estabilidade lipídica de linguiça tipo Toscana, comparados ao eritorbato de sódio. As cascas foram submetidas à extração e liofilização aquosas por ultrassom. O extrato foi caracterizado por determinação de fenol e flavonóides. Os parâmetros cor, pH, peróxidos e ácido tiobarbitúrico foram avaliadas nos dias 1, 15 e 30 pós armazenamento, a temperatura de 5 ± 1 ºC e ciclo de luz de 12 horas/dia. As linguiças contendo 0,05 e 0,1% de extrato de casca apresentaram resultados semelhantes ao eritorbato de sódio quanto a peroxidação lipídica. Também apresentaram aceitabilidade global adequada na análise sensorial. Os extratos de casca de romã são, portanto, uma alternativa natural promissora para manter a estabilidade lipídica das linguiças toscanas, promovendo a proteção e podendo trazer benefícios da romã à saúde humana.(AU)

Análise molecular para Toxoplasma gondii em linguiças suínas defumadas / Molecular analysis for Toxoplasma gondii in smoked swine sausages

A toxoplasmose é causada pelo protozoário Toxoplasma gondii, o qual acomete os humanos através da ingestão de carne mal passada ou crua. O presente trabalho descreve o emprego de PCR em linguiças suínas defumadas (salames) para detecção de T. gondii, aliado a utilização do teste de imunofluorescência indireta na avaliação sorológica de suínos encaminhados para abate. No estudo, avaliaram-se 18 amostras de salames e 50 amostras de soro sanguíneo de suínos. Na PCR todas as amostras de salames se apresentaram negativas e no teste de imunofluorescência indireta 8% dos animais foram positivos para T. gondii. Embora PCR-negativas, as linguiças produzidas originaram-se de matéria-prima suína proveniente de estabelecimento de abate, cujo presente estudo identificou soroprevalência de 8% para o protozoário. Dessa forma, o consumo de carne mal passada ou crua e de produtos a base de carne crua, como os salames, devem ser evitados, principalmente, em grupos de risco como crianças e idosos.(AU)

Tuberculose bovina em frigoríficos sob fiscalização estadual no Estado do Rio Grande do Sul nos anos de 2015 a 2017 / Bovine tuberculosis in slaughterhouse under state sanitary inspection in Rio Grande do Sul from 2015 to 2017

A vigilância em matadouros frigoríficos através da inspeção post mortem fornece um mecanismo importante para a detecção de infecções por tuberculose bovina (Mckinley et al 2018). As taxas de detecção de tuberculose bovina em matadouros frigoríficos dependem de fatores como eficácia da inspeção de produtos de origem animal bem como perfil de risco e susceptibilidade da população bovina abatida. Este trabalho teve por objetivo demonstrar a taxa de condenações por lesões sugestivas de Tuberculose bovina em estabelecimentos frigoríficos sob fiscalização Estadual no estado do Rio Grande do Sul entre janeiro de 2015 a dezembro de 2017. Nestes estabelecimentos foram abatidos em 2015 um total de 1.020.554 (100%) animais dos quais 3.622 (0,35%) condenados. Em 2016, 1.002.496 (100%) bovinos foram abatidos, e 5.587 (0,55%) foram condenados. Já em 2017 estes dados totalizaram 1.055.585 (100%) animais abatidos e 6.466 (0,61%) condenados. Os resultados demonstram um aumento gradativo da identificação de lesões sugestivas de tuberculose em matadouros frigoríficos no estado do RS nos anos estudados.(AU)

Tuberculose bovina: diagnóstico ante mortem e post mortem em bovinos leiteiros através da associação de análises / Bovine tuberculosis: diagnosis ante mortem and post mortem in dairy cattle through analysis association

Este estudo descreve o emprego de quatro métodos diagnósticos para a confirmação de casos de tuberculose em bovinos. Para tanto foram avaliados 211 bovinos leiteiros, dos quais 35,1% apresentaram reatividade no teste tuberculínico cervical comparativo, e 143 animais foram encaminhados para abate sanitário. No acompanhamento do abate e inspeção sanitária verificou-se que 51,8% dos bovinos apresentavam lesões visíveis compatíveis com tuberculose. Também foram coletados fragmentos teciduais de bovinos com e sem lesões macroscópicas, os quais na análise histológica apresentaram numerosas áreas de necrose caseosa e áreas de calcificação, e na coloração de Ziehl-Neelsen apresentaram bacilos álcool-ácido resistentes. Assim, as análises empregadas no estudo se mostraram importantes para o diagnóstico acurado de tuberculose bovina, além de alertar para o risco de saúde pública que pessoas que trabalham nos abatedouros e/ou em contato direto com bovinos doentes estão submetidas.(AU)

Análise molecular para Toxoplasma gondii em linguiças suínas defumadas / Molecular analysis for Toxoplasma gondii in smoked swine sausages

A toxoplasmose é causada pelo protozoário Toxoplasma gondii, o qual acomete os humanos através da ingestão de carne mal passada ou crua. O presente trabalho descreve o emprego de PCR em linguiças suínas defumadas (salames) para detecção de T. gondii, aliado a utilização do teste de imunofluorescência indireta na avaliação sorológica de suínos encaminhados para abate. No estudo, avaliaram-se 18 amostras de salames e 50 amostras de soro sanguíneo de suínos. Na PCR todas as amostras de salames se apresentaram negativas e no teste de imunofluorescência indireta 8% dos animais foram positivos para T. gondii. Embora PCR-negativas, as linguiças produzidas originaram-se de matéria-prima suína proveniente de estabelecimento de abate, cujo presente estudo identificou soroprevalência de 8% para o protozoário. Dessa forma, o consumo de carne mal passada ou crua e de produtos a base de carne crua, como os salames, devem ser evitados, principalmente, em grupos de risco como crianças e idosos.

Tuberculose bovina: diagnóstico ante mortem e post mortem em bovinos leiteiros através da associação de análises / Bovine tuberculosis: diagnosis ante mortem and post mortem in dairy cattle through analysis association

Este estudo descreve o emprego de quatro métodos diagnósticos para a confirmação de casos de tuberculose em bovinos. Para tanto foram avaliados 211 bovinos leiteiros, dos quais 35,1% apresentaram reatividade no teste tuberculínico cervical comparativo, e 143 animais foram encaminhados para abate sanitário. No acompanhamento do abate e inspeção sanitária verificou-se que 51,8% dos bovinos apresentavam lesões visíveis compatíveis com tuberculose. Também foram coletados fragmentos teciduais de bovinos com e sem lesões macroscópicas, os quais na análise histológica apresentaram numerosas áreas de necrose caseosa e áreas de calcificação, e na coloração de Ziehl-Neelsen apresentaram bacilos álcool-ácido resistentes. Assim, as análises empregadas no estudo se mostraram importantes para o diagnóstico acurado de tuberculose bovina, além de alertar para o risco de saúde pública que pessoas que trabalham nos abatedouros e/ou em contato direto com bovinos doentes estão submetidas.

Tuberculose bovina em frigoríficos sob fiscalização estadual no Estado do Rio Grande do Sul nos anos de 2015 a 2017 / Bovine tuberculosis in slaughterhouse under state sanitary inspection in Rio Grande do Sul from 2015 to 2017

A vigilância em matadouros frigoríficos através da inspeção post mortem fornece um mecanismo importante para a detecção de infecções por tuberculose bovina (Mckinley et al 2018). As taxas de detecção de tuberculose bovina em matadouros frigoríficos dependem de fatores como eficácia da inspeção de produtos de origem animal bem como perfil de risco e susceptibilidade da população bovina abatida. Este trabalho teve por objetivo demonstrar a taxa de condenações por lesões sugestivas de Tuberculose bovina em estabelecimentos frigoríficos sob fiscalização Estadual no estado do Rio Grande do Sul entre janeiro de 2015 a dezembro de 2017. Nestes estabelecimentos foram abatidos em 2015 um total de 1.020.554 (100%) animais dos quais 3.622 (0,35%) condenados. Em 2016, 1.002.496 (100%) bovinos foram abatidos, e 5.587 (0,55%) foram condenados. Já em 2017 estes dados totalizaram 1.055.585 (100%) animais abatidos e 6.466 (0,61%) condenados. Os resultados demonstram um aumento gradativo da identificação de lesões sugestivas de tuberculose em matadouros frigoríficos no estado do RS nos anos estudados.

Biofilm formation by Salmonella Enteritidis at different incubation temperatures

Background: The genus Salmonella, associated with poultry products, is considered the leading cause of foodborne outbreaks in humans in many countries. In Brazil, Salmonella Enteritidis (SE) is the serovar remains as one most frequentlyisolated from humans, and it is also a major serovar found in animals, food, animal feed, and environmental samples, despiteall the efforts to control this pathogen. Also, the bacterium is able to form biofilms on different surfaces, protecting cellsfrom both cleaning and sanitizing procedures in the food industries. This study aimed to verify the ability of SalmonellaEnteritidis isolates to form biofilm on polystyrene at different incubation temperatures.Materials, Methods & Results: A total of 171 SE samples were isolated from foodborne outbreaks (foods and stool cultures) and poultry products between 2003 and 2010. The biofilm-forming ability of samples was measured at four differenttemperatures (3°C, 9ºC, 25ºC, and 36ºC), for 24 h, simulating temperatures usually found in poultry slaughterhouses. Later,200 μL of each bacterial suspension was inoculated, in triplicate, onto 96-well, flat-bottomed sterile polystyrene microtiterplates, washed, after that, the biofilm was fixed with methanol. The plates were dried at ambient temperature, stained with2% Hucker’s crystal violet. Afterwards, absorbance was read using an ELISA plate reader and the optical density (OD)of each isolate was obtained by the arithmetic mean of the absorbance of three wells and this value was compared withthe mean absorbance of negative controls (ODnc). The following classification was used for the determination of biofilmformation: no biofilm production, weak biofilm production, moderate biofilm production and strong biofilm production.Results demonstrated all isolates from stool cultures and foods involved in foodborne outbreaks, at least one of the fourtemperatures tested, were able to form biofilm...

Quantificação de Staphylococcus aureus e bactérias mesófilas aeróbias para avaliar higienização de equipamentos de ordenha / Staphylococcus aureus and mesophilic aerobic bacteria quantification in hygienization process of milking equipment

Background: Milk’s composition is an excellent substrate for microorganism’s multiplication. Presence of Staphylococcusaureus and aerobic mesophilic bacteria are one of the most common problems in dairy farms. On dairy industry’s and milkfarms Clean in Place (CIP) system higyenization are commonly used, then the cleaning occurs as a closed process, forbetter results sanitizans are applied, in order to obtain a safety food. This project aim to evaluate Staphylococcus aureusand aerobic mesophilic bacteria reduction after two milking higyenization process.Materials, Methods & Results: This research was done on a Rio Grande do Sul North Milk farm, with mechanized milkingand Clean in Place system for cleaning. For liners and CIP tubes higyenization commercial products as Sodium Hipoclorite3% and phosphoric acid 11.3% are used for detergency, and peracetic acid 5% for sanitization. Milk bunk tank are higyenized with sodium hypoclorite 3.8% alcalin detergent. After higyenization steps liners, CIP’s water process, bulk milk tankand milk set were collected. At process 1, liners and CIP water were collected after milking, detergency and sanitizationthat occurred immediately at the detergency’s finish, while process 2 the sanitization was realized 8 h after detergency,before following milking. Cooling milk bulk tank was collected before and after detergency, and milk set after milkingsConvencional microbiology were used to count and results in log10 UFC.cm-2...

Biofilm formation by Salmonella Enteritidis at different incubation temperatures

Background: The genus Salmonella, associated with poultry products, is considered the leading cause of foodborne outbreaks in humans in many countries. In Brazil, Salmonella Enteritidis (SE) is the serovar remains as one most frequentlyisolated from humans, and it is also a major serovar found in animals, food, animal feed, and environmental samples, despiteall the efforts to control this pathogen. Also, the bacterium is able to form biofilms on different surfaces, protecting cellsfrom both cleaning and sanitizing procedures in the food industries. This study aimed to verify the ability of SalmonellaEnteritidis isolates to form biofilm on polystyrene at different incubation temperatures.Materials, Methods & Results: A total of 171 SE samples were isolated from foodborne outbreaks (foods and stool cultures) and poultry products between 2003 and 2010. The biofilm-forming ability of samples was measured at four differenttemperatures (3°C, 9ºC, 25ºC, and 36ºC), for 24 h, simulating temperatures usually found in poultry slaughterhouses. Later,200 μL of each bacterial suspension was inoculated, in triplicate, onto 96-well, flat-bottomed sterile polystyrene microtiterplates, washed, after that, the biofilm was fixed with methanol. The plates were dried at ambient temperature, stained with2% Huckers crystal violet. Afterwards, absorbance was read using an ELISA plate reader and the optical density (OD)of each isolate was obtained by the arithmetic mean of the absorbance of three wells and this value was compared withthe mean absorbance of negative controls (ODnc). The following classification was used for the determination of biofilmformation: no biofilm production, weak biofilm production, moderate biofilm production and strong biofilm production.Results demonstrated all isolates from stool cultures and foods involved in foodborne outbreaks, at least one of the fourtemperatures tested, were able to form biofilm...(AU)

Quantificação de Staphylococcus aureus e bactérias mesófilas aeróbias para avaliar higienização de equipamentos de ordenha / Staphylococcus aureus and mesophilic aerobic bacteria quantification in hygienization process of milking equipment

Background: Milks composition is an excellent substrate for microorganisms multiplication. Presence of Staphylococcusaureus and aerobic mesophilic bacteria are one of the most common problems in dairy farms. On dairy industrys and milkfarms Clean in Place (CIP) system higyenization are commonly used, then the cleaning occurs as a closed process, forbetter results sanitizans are applied, in order to obtain a safety food. This project aim to evaluate Staphylococcus aureusand aerobic mesophilic bacteria reduction after two milking higyenization process.Materials, Methods & Results: This research was done on a Rio Grande do Sul North Milk farm, with mechanized milkingand Clean in Place system for cleaning. For liners and CIP tubes higyenization commercial products as Sodium Hipoclorite3% and phosphoric acid 11.3% are used for detergency, and peracetic acid 5% for sanitization. Milk bunk tank are higyenized with sodium hypoclorite 3.8% alcalin detergent. After higyenization steps liners, CIPs water process, bulk milk tankand milk set were collected. At process 1, liners and CIP water were collected after milking, detergency and sanitizationthat occurred immediately at the detergencys finish, while process 2 the sanitization was realized 8 h after detergency,before following milking. Cooling milk bulk tank was collected before and after detergency, and milk set after milkingsConvencional microbiology were used to count and results in log10 UFC.cm-2...(AU)

Salmonella Enteritidis forms biofilm under low temperatures on different food industry surfaces / Salmonella Enteritidis forma biofilme sob baixas temperaturas em diferentes superfícies da indústria de alimentos

We evaluated the influence of temperature on the ability of Salmonella Enteritidis (SE) to form biofilms on stainless steel, polyethylene, and polyurethane surfaces under different hygiene procedures. These materials were placed on SE culture and incubated at 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC, and 3±1 ºC for 4, 8, 12, and 24 h. Hot water at 45 ºC and 85 ºC, 0.5% peracetic acid solution, and 1% quaternary ammonia were used for hygienization. Biofilm formation occurred at all temperatures evaluated, highlighting at 3 ºC which has not been reported as an ideal temperature for the adhesion of SE to these materials. The SE adhered more often to polyethylene surfaces than to polyurethane and stainless steel surfaces (P<0.05). Peracetic acid and water at 85 ºC had similar hygienization efficiency (P<0.05) followed by quaternary ammonia whereas water at 45 ºC was not effective. SE adhered to these materials under low temperatures which to date have been deemed safe for food preservation.(AU)

Avaliou-se o efeito da temperatura na capacidade de Salmonella Enteritidis (SE) formar biofilme em superfícies de aço inoxidável, polietileno e poliuretano e diferentes processos de higienização. Corpos de prova destes materiais foram postos frente a culturas de SE e incubados a 42±1 ºC, 36±1 ºC, 25±1 ºC, 9±1 ºC e 3±1 ºC por 4, 8, 12 e 24 horas. Para a higienização foram testados água aquecida a 45ºC e 85 ºC e soluções de ácido peracético 0,5% e amônia quaternária 1%. Verificou-se a formação de biofilmes em todas as temperaturas avaliadas, ressaltando-se a 3 ºC, ainda não citada como propícia para adesão de SE. Houve maior adesão ao polietileno do que ao poliuretano e ao aço inoxidável (P<0.05). Para higienização, o ácido peracético e a água a 85 ºC tiveram ação semelhante (P<0.05), seguidos por amônia quaternária, enquanto que a água a 45 ºC não foi eficaz. Todos os materiais avaliados propiciaram a aderência de SE, mesmo sob temperaturas baixas, consideradas até então seguras para a conservação dos alimentos.(AU)

ATP-Bioluminescence and conventional microbiology for hygiene evaluation of cutting room surfaces in poultry slaughterhouse

Background: The hygiene procedures in poultry slaughterhouses consist in the use of hot water, detergent and sanitizing, configuring Sanitation Standard Operating Procedure (SSOP). These actions control contamination in food processing environments, especially by pathogenic microorganisms, which cause diseases with impact on public health and economic losses. The microbiological control of aerobic mesophiles, Staphylococcus aureus and Escherichia coli, are used as indicators of contamination. The hygienic-sanitary conditions on the surfaces of the poultry slaughterhouse cuttting room were evaluated, before and after cleaning and sanitizing procedures.Materials, Methods & Results: Conventional microbiology (Rodac plates and sponge for quantification of aerobic mesophiles, Staphylococcus aureus and Escherichia coli) and ATP-Bioluminescence were used to analyze the action of hot water and the active principles peracetic acid, quaternary ammonia and biguanide in the standard pre-operational hygiene procedure in the cutting room of the poultry slaughterhouse under Federal Inspection with slaughter capacity of more than 20.000 birds/h. The evaluations were performed on three lines of chicken thigh cuts at the same time and in a completely randomized manner on stainless steel surfaces, polyurethane belts and polyethylene boards. Samples were made in four replicates at the three surface totaling 108 assay for each microorganism. The samples were collected at the end of the cutting process, before and after washing the surfaces with hot water (between 45 and 50ºC) and after sanitization with 0.5% peracetic acid, 2% quaternary ammonia and 1% biguanide. The ATP-Bioluminescence method detected organic matter at all collected points and Rodac plates allowed a better recovery of microorganisms than sponges for quantification of aerobic mesophiles, E. coli and S. aureus.[...]

ATP-Bioluminescence and conventional microbiology for hygiene evaluation of cutting room surfaces in poultry slaughterhouse

Background: The hygiene procedures in poultry slaughterhouses consist in the use of hot water, detergent and sanitizing, configuring Sanitation Standard Operating Procedure (SSOP). These actions control contamination in food processing environments, especially by pathogenic microorganisms, which cause diseases with impact on public health and economic losses. The microbiological control of aerobic mesophiles, Staphylococcus aureus and Escherichia coli, are used as indicators of contamination. The hygienic-sanitary conditions on the surfaces of the poultry slaughterhouse cuttting room were evaluated, before and after cleaning and sanitizing procedures.Materials, Methods & Results: Conventional microbiology (Rodac plates and sponge for quantification of aerobic mesophiles, Staphylococcus aureus and Escherichia coli) and ATP-Bioluminescence were used to analyze the action of hot water and the active principles peracetic acid, quaternary ammonia and biguanide in the standard pre-operational hygiene procedure in the cutting room of the poultry slaughterhouse under Federal Inspection with slaughter capacity of more than 20.000 birds/h. The evaluations were performed on three lines of chicken thigh cuts at the same time and in a completely randomized manner on stainless steel surfaces, polyurethane belts and polyethylene boards. Samples were made in four replicates at the three surface totaling 108 assay for each microorganism. The samples were collected at the end of the cutting process, before and after washing the surfaces with hot water (between 45 and 50ºC) and after sanitization with 0.5% peracetic acid, 2% quaternary ammonia and 1% biguanide. The ATP-Bioluminescence method detected organic matter at all collected points and Rodac plates allowed a better recovery of microorganisms than sponges for quantification of aerobic mesophiles, E. coli and S. aureus.[...](AU)

Economic losses associated with Wooden Breast and White Striping in broilers / Perdas econômicas associadas as alterações Wooden Breast e White Striping em frangos de corte

Currently, two defects in poultry breasts termed Wooden Breast (WB) and White Striping (WS) havebeen reported in slaughterhouses. These defects may be associated with the accelerated growth of thebirds, management, density in the aviary, and both weight and age at slaughter. Although the health ofthe affected birds is not impaired, these myopathies cause carcass condemnation and economic losses toslaughterhouses, since the breasts of these chickens, considered to be prime cuts in the poultry industry,have to be discarded. This paper reports on the economic losses and factors associated with carcasscondemnation caused by white striping (WS) and wooden breast (WB) in broilers from a federallyinspected slaughterhouse. Twelve flocks, totaling 207,000 slaughtered broilers, were assessed as toweight and age at slaughter, and stocking density and carcass condemnations due to these two myopathieswere also evaluated. Economic losses were estimated by the price of poultry breast at the firm level,around U$ 19,12 per kilo, amounting to daily losses of up to U$ 70,632.00, given that approximately0.8% of the chicken breasts were condemned. Heavier broilers had a larger condemnation rate due toWS and WB, and so did those reared at a smaller density, due probably to their better access to waterand feed, which contributed to weight gain and consequent condemnation, resulting in losses that couldaffect the entire poultry sector.

Atualmente, dois defeitos em peitos de frango, denominados Peito Madeira (Wooden Breast - WB)e Peito com Estrias Brancas (White Striping- WS) têm sido relatados em abatedouros e podem estarassociados ao crescimento acelerado das aves, manejo, densidade no aviário, peso e idade ao abate.Embora não haja comprometimento da saúde das aves afetadas, estas miopatias causam condenações eprejuízos aos abatedouros devido ao não aproveitamento do peito destes frangos, considerado um cortenobre na indústria avícola. Neste trabalho são relatados os prejuízos econômicos e fatores associados àscondenações por White Striping (WS) e Wooden Breast (WB) em frangos de corte em um abatedourosob Inspeção Federal. Foram acompanhados 12 lotes totalizando 207.000 aves abatidas, registrando-se dados como peso e idade ao abate, densidade nos galpões de criação e condenações por estes doisdefeitos. As perdas econômicas foram calculadas a partir do valor do peito de frango na empresa, emtorno de R$ 5,90 o quilograma, gerando prejuízos de até R$ 21.800,00 ao dia, já que cerca de 0,8%dos peitos foram desqualificados devido à estas alterações. Aves mais pesadas apresentaram maiorcondenação por WS e WB, bem como quando criadas em menor densidade, provavelmente devido aomelhor acesso a água e ração, contribuindo para aumento de peso e consequente condenações por estasmiopatias, impactando em prejuízos que podem ser extrapolados para todo o setor avícola.