Salmonella Serovars Isolated from Poultry Breeding Flocks under the Brazilian Official Control Programme Between 2016 and 2018

The control of Salmonella in the poultry production chain combined with biosecurity measures is an important tool to maintain and guarantee the sanitary status of Brazilian flocks. The aim of this work was to compare official laboratory data on molecular typification of Salmonella isolates from poultry breeding flocks in different Brazilian states between 2016 and 2018 and identify the production category with the most positive flocks, in light of current legislation. Surveillance data of positive samples from the official Brazilian Salmonella Control Programme sent to Federal Agricultural Defence Laboratory of São Paulo (LFDA-SP) after molecular characterization were analysed. These data were subject to an exploratory study, undergoing a descriptive statistical analysis followed by the use of frequency and non-parametric hypothesis tests. Overall, 49 serovars were detected in poultry broiler-breeder and layer-breeder flocks. Salmonella ser. Heidelberg, Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Schwarzengrund and Salmonella ser. Mbandaka were the five most common isolated serovars. The data shows that there is an opportunity to improve biosecurity measures in parent breeder flocks. A total of 16 serovars were identified in turkey-breeders. Salmonella ser. Anatum, Salmonella ser. Newport, Salmonella ser. Brandenburg, Salmonella ser. Litchfield, and Salmonella ser. Livingstone were the most common ones. The four official controlled serovars represented a small part of the isolated strains. These data demonstrate the importance of an official program in Brazil for Salmonella surveillance in breeder flocks combined with biosecurity measures.(AU)

Acid resistance of Salmonella Typhimurium ATCC 14028 after desiccation stress during peanut storage / Avaliação da resistência ácida de Salmonella Typhimurium ATCC 14028 após o estresse dessecativo durante o armazenamento de amendoim

Salmonella is a relevant pathogen, which causes foodborne outbreaks associated with both high and low moisture foods (LMF). This study evaluated the effect of previous desiccation stress on the acid resistance of S. Typhimurium ATCC 14028 using blanched peanut kernels as an LMF model. Salmonella was recovered from the peanut samples throughout 180 days of blanched peanut kernels storage at 28 ºC. During this period two death rates were verified, 0.04 log cfu/g/day in the first 30 days and 0.007 log cfu/g/day between 30 and180 days.Regarding acid resistance, there was no difference (P > 0.05) in the Salmonella growth/death kinetics between the undesiccated sample (TSB)and the cells recovered from peanut samples over 180 days of storage after 4 h at pHs 3.0, 3.5, 4.5 and 7.2. The average growth rate observed for pH 7.2 was 0.44 log cfu/ml/h. At pH 4.5, the Salmonella counts did not change significantly over 4 h. In contrast, Salmonella populations declined by 0.14 to 0.29 log cfu/ml/h at pH 3.5. At pH 3.0 declines were estimated to be 0.65 log cfu/ml/h for the undesiccated sample and 2.07 log cfu/ml/h for Salmonella recovered from peanuts stored for 120 days. Therefore, our data indicated that desiccation stress caused during the peanut storagedid not influence the Salmonella acid resistance.

Em alimentos de baixa umidade (LMF), o primeiro desafio encontrado por patógenos como a Salmonella é o estresse de dessecação. Neste estudo, o efeito prévio do estresse de dessecação sobre a resistência ácida de S. Typhimurium ATCC 14028 foi avaliado utilizando amendoim blancheado como um modelo de LMF. Salmonella foi recuperada das amostras de amendoim após 180 dias de estocagem a 28 ºC. Durante este período foram verificadas duas taxas de mortalidade, 0,04 log ufc/g/dia nos primeiros 30 dias e 0,007 log ufc/g/dia entre 30 e 180 dias. Com relação à resistência ácida, não houve diferença (P > 0,05) na cinética de crescimento/morte de Salmonella entre a amostra sem estresse dessecativo (TSB) e as amostras de amendoim após 4 h em pHs 3,0, 3,5, 4,5 e 7,2. A taxa média de crescimento observada para o pH 7,2 foi de 0,44 log cfu/ml/h. No pH 4,5, a contagem de Salmonella não mudou significativamente durante 4 h. Em contraste, a população de Salmonella diminuiu de 0,14 a 0,29 log cfu/ml/h no pH 3,5. No pH 3,0, a queda foi estimada em 0,65 log cfu/ml/h para a amostra sem estresse dessecativo e 2,07 log cfu/ml/h para a Salmonella recuperada das amostras de amendoim estocadas por 120 dias. Portanto, nossos dados indicaram que o estresse dessecativo causado durante a estocagem do amendoim não influenciou a resistência ácida de Salmonella.

Effect of Ammonia Gas in Poultry Litter Contaminated with Salmonella Heidelberg

Salmonella Heidelberg is an emerging pathogen in Brazilian poultry production. The traditional methods (quicklime, windrowing and tarpaulin-on-surface) used for disinfecting reused poultry litter between flocks does not guarantee its elimination, thus allowing the transmission of this agent from one flock to another. The new tarpaulinon-surface method with controlled injection of ammonia gas has proven to be effective in its control, however, it is still unknown what dose of ammonia gas is needed to eliminate Salmonella Heidelberg in reused poultry litter. The objective of this study was to evaluate the effect of ammonia gas at different concentrations in sterile poultry litter artificially contaminated with Salmonella Heidelberg. Then, ammonia gas was injected in concentrations of 0.25%, 0.5%, and 1%, and 48 hours later, a sample was collected from each repetition in an entirely randomized design, and bacterial isolation was performed. All treatments, including positive and negative controls, were tested in quadruplicate and the parameters temperature, humidity, pH and water activity were evaluated. In the 0.5% and 1% treated samples the pathogen was not isolated, while in the 0.25% concentration one of the four samples tested was positive. The study reveals that ammonia gas is efficient in killing Salmonella Heidelberg in poultry litter at concentrations of 0.5 % or more within a 48-hour period and that the litter treated with ammonia gas increases its pH and water activity.(AU)

Acid resistance of Salmonella Typhimurium ATCC 14028 after desiccation stress during peanut storage / Avaliação da resistência ácida de Salmonella Typhimurium ATCC 14028 após o estresse dessecativo durante o armazenamento de amendoim

ABSTRACT: Salmonella is a relevant pathogen, which causes foodborne outbreaks associated with both high and low moisture foods (LMF). This study evaluated the effect of previous desiccation stress on the acid resistance of S. Typhimurium ATCC 14028 using blanched peanut kernels as an LMF model. Salmonella was recovered from the peanut samples throughout 180 days of blanched peanut kernels storage at 28 ºC. During this period two death rates were verified, 0.04 log cfu/g/day in the first 30 days and 0.007 log cfu/g/day between 30 and180 days.Regarding acid resistance, there was no difference (P > 0.05) in the Salmonella growth/death kinetics between the undesiccated sample (TSB)and the cells recovered from peanut samples over 180 days of storage after 4 h at pHs 3.0, 3.5, 4.5 and 7.2. The average growth rate observed for pH 7.2 was 0.44 log cfu/ml/h. At pH 4.5, the Salmonella counts did not change significantly over 4 h. In contrast, Salmonella populations declined by 0.14 to 0.29 log cfu/ml/h at pH 3.5. At pH 3.0 declines were estimated to be 0.65 log cfu/ml/h for the undesiccated sample and 2.07 log cfu/ml/h for Salmonella recovered from peanuts stored for 120 days. Therefore, our data indicated that desiccation stress caused during the peanut storagedid not influence the Salmonella acid resistance.

RESUMO: Em alimentos de baixa umidade (LMF), o primeiro desafio encontrado por patógenos como a Salmonella é o estresse de dessecação. Neste estudo, o efeito prévio do estresse de dessecação sobre a resistência ácida de S. Typhimurium ATCC 14028 foi avaliado utilizando amendoim blancheado como um modelo de LMF. Salmonella foi recuperada das amostras de amendoim após 180 dias de estocagem a 28 ºC. Durante este período foram verificadas duas taxas de mortalidade, 0,04 log ufc/g/dia nos primeiros 30 dias e 0,007 log ufc/g/dia entre 30 e 180 dias. Com relação à resistência ácida, não houve diferença (P > 0,05) na cinética de crescimento/morte de Salmonella entre a amostra sem estresse dessecativo (TSB) e as amostras de amendoim após 4 h em pHs 3,0, 3,5, 4,5 e 7,2. A taxa média de crescimento observada para o pH 7,2 foi de 0,44 log cfu/ml/h. No pH 4,5, a contagem de Salmonella não mudou significativamente durante 4 h. Em contraste, a população de Salmonella diminuiu de 0,14 a 0,29 log cfu/ml/h no pH 3,5. No pH 3,0, a queda foi estimada em 0,65 log cfu/ml/h para a amostra sem estresse dessecativo e 2,07 log cfu/ml/h para a Salmonella recuperada das amostras de amendoim estocadas por 120 dias. Portanto, nossos dados indicaram que o estresse dessecativo causado durante a estocagem do amendoim não influenciou a resistência ácida de Salmonella.

Three Months of Composting Are Enough to Eliminate the Fowl Typhoid Bacteria

The composting technique has been increasingly highlighted in poultry production units, as an efficient and low-cost solution for the destination of carcasses. The process is based on the accelerated decomposition of organic material under high temperatures, associated with eliminating pathogenic microorganisms. This study aims to evaluate the effectiveness and the time necessary for the elimination of Salmonella Gallinarum in carcasses of poultry submitted to the composting process. The composting was carried out following the models used in the field, and microbiological analysis was performed in five different periods: 45, 90, 120, 150 and 180-days after closing the composter. After 90 days of experiment and in the subsequent analysis, the elimination of the bacteria in 100% of the samples was verified, validating the composting process as an effective method for eliminating S. Gallinarum in poultry carcasses, when respecting the period necessary for the elimination of the bacteria and the good quality of the structure adopted for the process.(AU)

Use of organic acids to reduce Salmonella Typhimurium excretion in swine / Uso de ácidos orgânicos para reduzir a excreção de Salmonella Typhimurium em suínos

The use of antimicrobials as growth promoters and disease prevention is being constantly reduced in several animal production systems, including in the swine industry. Therefore, this study aimed to evaluate the effectiveness of using acidifiers to control Salmonella Typhimurium in 65-day-old pigs by detecting the pathogen in organs at euthanasia. For this, 24 piglets were divided into two experimental groups consisting of 12 piglets each. An untreated control group (G1) and a treatment group (G2) received a liquid organic acidifier in the drinking water for 10 days (D-5 to D5). Five days after the start of treatment (D0), all piglets were challenged with 106 CFU of Salmonella Typhimurium and assessed for 12 days (D12). Every three days (D3, D6, D9, and D12), three animals from each experimental group were euthanized and then submitted for necropsy. Samples from the intestines (ileum, cecum, mesenteric lymph nodes, and ileocolic lymph nodes), liver, spleen, and lungs were collected to isolate Salmonella. The results show that, numerically, Salmonella isolation in the organs of G2 was lower than in G1 and that the number of positive cecum samples in G1 (66.7%; 8/12) was statistically different from the number of positive models in G2 (16.7%; 2/12), with a reduction of 28.6% of the total cecum positive samples in the treated group compared to the control. Therefore, it was observed that the liquid organic acidifier product could reduce the colonization of organs by Salmonella Typhimurium. (AU)

A Comparative Study of the Effect of Non-Antibiotic Feed Additives on Experimental Colonization of Salmonella Enterica Serovar Enteridis and Intestinal Pathomorphology in Broiler Chickens

The objective of this study was to evaluate the effect of eubiotics on the intestinal morphology of broilers. For this purpose, 125 birds were divided into six groups with two replicates each (10 birds in each replicate). Group A was given a Basal diet. All groups except group A were challenged with Salmonella enterica serovar Enteritidis. Group B was provided the basal diet, group C was fed a Probiotic-added diet; group D was fed a Prebiotics-based diet; group E was given essential oils plus the basal diet; and group F was provided with organic acids plus the basal diet. Two separate experiments were carried out for Salmonella recovery, checking the cecal tonsils and conducting an intestinal pathomorphic evaluation. Villus length, villus width, villus surface area, and crypt depth were measured by micrometry. There was an overall improvement (p<0.05) in intestinal morphometric parameters for all the treatment groups except for the negative control group, which showed the lowest villus height and villus depth values. Maximum villus height (p<0.05) of the duodenum was achieved by group E, which was fed a diet containing essential oils, whereas a maximum villus surface area index (p<0.05) was recorded for the birds of Group D, which were fed a diet containing prebiotics. Maximum villus height (p<0.05) and surface area index in ileum mucosa was recorded (p<0.05) in the birds of group D (treated with prebiotics). It is concluded that there is an overall increase in the gut histology of broilers fed non-antibiotic based feed.(AU)

An outbreak of fatal Pullorum disease (Salmonella Pullorum) in Guinea fowl keets (Numida meleagris) / Surto fatal de pulorose (Salmonella Pullorum) em pintinhos de galinhas-d'Angola (Numida meleagris)

Pullorum disease is described worldwide and is caused by Salmonella enterica subspecies enterica serovar Gallinarum biovar Pullorum (S. Pullorum). S. Pullorum infection is important in commercial poultry, provoking a systemic disease with high mortality rates. Its occurrence requires notification, and when it is diagnosed in commercial breeding flocks, its eradication is demanded. The aim of this study was to report a severe outbreak of Pullorum disease in young Guinea fowl (Numida meleagris), resulting in 100% mortality of keets (n=290) within the first two weeks of age. All examined keets had enlarged liver, kidneys and spleen (5/5), and the affected tissues were submitted to histological and bacteriological examination. On histopathology, random paratyphoid nodules characterized by areas of necrosis with fibrin and a moderate infiltrate of macrophages and heterophils were observed in the liver. In kidneys, discrete areas of necrosis associated with moderate multifocal infiltrates of lymphocytes, and plasma cells were observed. In the spleen, a moderate infiltrate of macrophages was noticed. Isolation of colonies suggestive of S. Pullorum from liver and spleen was performed in selective agars and, after biochemical tests, confirmed by specific duplex-PCR. The antimicrobial susceptibility test of the isolated strain revealed resistance to only sulfamethoxazole + trimethoprim among the tested antimicrobials. The S. Pullorum isolate recovered in the present study was highly pathogenic to N. meleagris and may represent a risk to other avian species, including industrial poultry.

A pulorose é descrita mundialmente e é causada por Salmonella enterica subespécie enterica sorovar Gallinarum biovar Pullorum (S. Pullorum). A infecção por S. Pullorum é importante em aves comerciais, provocando doença sistêmica com altas taxas de mortalidade. Sua ocorrência requer notificação e quando diagnosticada em aves de criação comercial resulta na erradicação do plantel. O objetivo deste estudo foi relatar um surto grave de pulorose em filhotes de galinhas-d'Angola (Numida meleagris), resultando em 100% de mortalidade das aves (n=290) nas primeiras duas semanas de idade. Os pintinhos recebidos tinham hepato, espleno e nefromegalia (5/5). Os tecidos dos cinco indivíduos recebidos foram submetidos a exame histológico e bacteriológico. Na histopatologia, foram observados nódulos paratifoides aleatórios caracterizados por áreas de necrose com fibrina e infiltrado moderado de macrófagos e heterófilos no fígado. Nos rins, foram observadas áreas discretas de necrose associadas a infiltrados multifocais moderados de linfócitos e plasmócitos. No baço, foi observado infiltrado moderado de macrófagos. O isolamento de colônias sugestivas de S. Pullorum de fígados e baços foi realizado em ágares seletivos e, após testes bioquímicos, confirmado por duplex-PCR específico. A susceptibilidade antimicrobiana da cepa isolada revelou resistência apenas ao sulfametoxazol + trimetoprim entre os antimicrobianos testados. O isolado de S. Pullorum recuperado no presente estudo foi altamente patogênico para N. meleagris e pode representar um risco para outras espécies de aves, incluindo aves industriais.

High Genetic Similarity Among Salmonella Heidelberg Isolated from Poultry Farms, Wild Animals, Beef, Poultry and Pork Meat, and Humans in Brazil

Salmonellosis is an important gastrointestinal infection in humans and cause of foodborne outbreaks in the world. In this context, molecular characterization is essential to understand how the strains circulate. The aim of this study was to evaluate the genotypic distribution of S. Heidelberg according to the source of isolation. The genetic relatedness of the S. Heidelberg isolates was determined by pulsed-field gel electrophoresis (PFGE). The most prevalent pulsotypes of cluster A were BRJF6X01.006 (27/95 = 28,42%) related between 1995 and 2011 in broilers, poultry meat and poultry farms, meat product and human, and BRJF6X01.001 (21/95 = 22,10%) related between 2011 and 2017 in wild animals, broilers, poultry meat, poultry farms, meat product, animal feed, and pork meat. The pulsotype BRJF6X01.001 shows a high distribution in the environmental and productive chain. The degree of similarity between pulsotypes BRJF6X01.006 and BRJF6X01.001 is 88%. To ensure the safety of human and animal health, holistic approaches, including surveillance of Salmonella throughout the environment and in the production chain, together with control measures, are critical. As transmission of Salmonella from food producing animals to wildlife and to the environment is considered potential public health problem, information on the survival and persistence of Salmonella in the environment and in potential reservoirs is of considerable importance.(AU)

Biosseguridade na avicultura de corte: impactos na produção e alternativas para prevenção de doenças / Biosecurity in beef poultry: impacts on production and alternatives for disease prevention / Bioseguridad en avicultura: impactos en la producción y alternativas para la prevención de enfermedades

A avicultura de corte levou ao Brasil a ser o no líder exportador de carne de frango, desde 2011, e o terceiro produtor global desta proteína. Portanto, é importante que todo produtor possua e mantenha um programa de biosseguridade continuado, respeitando rigorosamente cada etapa ou prática de manejo a fim de obter o sucesso econômico de sua produção. Sustentado pela medicina veterinária preventiva, um programa de biosseguridade deve apresentar aspectos direcionados a cada sistema de proteção em particular, para prevenir e controlar a presença e/ou introdução de microrganismos patogênicos nos rebanhos. O objetivo deste trabalho e apresentar uma revisão atualizada de literatura sobre programas de biosseguridade para evitar a proliferação de agentes patogênicos na avicultura de corte como os dois tipos de Salmonella que causam riscos à saúde pública e à dos animais. A pesquisa é qualitativa de cunho exploratório bibliográfico-documental, com pesquisa em sites como o Google Acadêmico, da revista de veterinária da Unipar, SCIELO, portal CAPES e sites governamentais. O resultado da pesquisa apresentou um panorama real sobre emprego de programas de biosseguridade no Brasil, direcionados à avicultura de corte, demonstrando que os produtores estão se conscientizando sobre a importância destes programas, devido à pressão do mercado exportador global. Conclui-se que ainda falta uma maior conscientização por parte de todos os produtores brasileiros, para evitar que o plantel produzido seja contaminado por agentes patogênicos, principalmente a Salmonella, evitando que a saúde pública e animal esteja comprometida. Somente desta maneira, o Brasil conseguirá manter e expandir mais o mercado avícola a nível global.(AU)

Poultry farming has led Brazil to be the leading exporter of chicken meat, since 2011, and the third global producer of this protein. Therefore, it is important that every producer has and maintains a continuous biosecurity program, strictly respecting each stage or management practice in order to obtain the economic success of their production. Supported by preventive veterinary medicine, a biosecurity program must present aspects directed to each protection system in particular, to prevent and control the presence and/or introduction of pathogenic microorganisms in herds. The objective of this work is to present an updated review of the literature on biosecurity programs to prevent the proliferation of pathogenic agents in poultry farming, such as the two types of Salmonella that pose risks to public and animal health. The research is qualitative, bibliographical-documentary exploratory, with research on sites such as Google Scholar, Unipar's veterinary magazine, SCIELO, CAPES portal and government sites. The result of the research presented a real panorama on the use of biosecurity programs in Brazil, directed to poultry production, demonstrating that producers are becoming aware of the importance of these programs, due to the pressure of the global export market. It is concluded that there is still a lack of greater awareness on the part of all Brazilian producers, to prevent the produced herd from being contaminated by pathogenic agents, mainly Salmonella, preventing public and animal health from being compromised. Only in this way will Brazil be able to maintain and further expand the poultry market at a global level.(AU)

La avicultura llevó a Brasil a ser el principal exportador de carne de pollo, desde 2011, y el tercer productor mundial de esta proteína. Por ello, es importante que todo productor cuente y mantenga un programa de bioseguridad continuo, respetando estrictamente cada etapa o práctica de manejo para obtener el éxito económico de su producción. Apoyado en la medicina veterinaria preventiva, un programa de bioseguridad debe presentar aspectos dirigidos a cada sistema de protección en particular, para prevenir y controlar la presencia y/o introducción de microorganismos patógenos en los rebaños. El objetivo de este trabajo es presentar una revisión actualizada de la literatura sobre programas de bioseguridad para prevenir la proliferación de agentes patógenos en la avicultura, como los dos tipos de Salmonella que presentan riesgos para la salud pública y animal. La investigación es cualitativa, bibliográfico-documental exploratoria, con pesquisa en sitios como Google Scholar, revista veterinaria de la Unipar, SCIELO, portal de la CAPES y sitios gubernamentales. El resultado de la investigación presentó un panorama real sobre el uso de programas de bioseguridad en Brasil, dirigidos a la producción avícola, demostrando que los productores están tomando conciencia de la importancia de estos programas, debido a la presión del mercado mundial de exportación. Se concluye que aún falta una mayor conciencia por parte de todos los productores brasileños, para evitar que el hato producido sea contaminado por agentes patógenos, principalmente Salmonella, evitando que se comprometa la salud pública y animal. Solo así Brasil podrá mantener y expandir aún más el mercado avícola a nivel mundial.(AU)

Use of organic acids to reduce Salmonella Typhimurium excretion in swine / Uso de ácidos orgânicos para reduzir a excreção de Salmonella Typhimurium em suínos

The use of antimicrobials as growth promoters and disease prevention is being constantly reduced in several animal production systems, including in the swine industry. Therefore, this study aimed to evaluate the effectiveness of using acidifiers to control Salmonella Typhimurium in 65-day-old pigs by detecting the pathogen in organs at euthanasia. For this, 24 piglets were divided into two experimental groups consisting of 12 piglets each. An untreated control group (G1) and a treatment group (G2) received a liquid organic acidifier in the drinking water for 10 days (D-5 to D5). Five days after the start of treatment (D0), all piglets were challenged with 106 CFU of Salmonella Typhimurium and assessed for 12 days (D12). Every three days (D3, D6, D9, and D12), three animals from each experimental group were euthanized and then submitted for necropsy. Samples from the intestines (ileum, cecum, mesenteric lymph nodes, and ileocolic lymph nodes), liver, spleen, and lungs were collected to isolate Salmonella. The results show that, numerically, Salmonellaisolation in the organs of G2 was lower than in G1 and that the number of positive cecum samples in G1 (66.7%; 8/12) was statistically different from the number of positive models in G2 (16.7%; 2/12), with a reduction of 28.6% of the total cecum positive samples in the treated group compared to the control. Therefore, it was observed that the liquid organic acidifier product could reduce the colonization of organs by Salmonella Typhimurium.(AU)

O uso de antimicrobianos como promotores de crescimento e prevenção de doenças vem sendo constantemente reduzido em diversos sistemas de produção animal, inclusive na suinocultura. Portanto, o objetivo do presente estudo foi avaliar a eficácia do uso de acidificantes no controle de Salmonella Typhimurium em suínos de 65 dias de idade, detectando o patógeno em órgãos após a eutanásia. Para isso, 24 leitões foram divididos em dois grupos experimentais constituídos por 12 leitões cada. Um grupo controle não tratado (G1) e um grupo de tratamento (G2) que recebeu um acidificante orgânico líquido na água de beber por 10 dias (D-5 a D5). Cinco dias após o início do tratamento (D0), todos os animais foram inoculados oralmente com 106 UFC de Salmonella Typhimurium e avaliados por 12 dias (D12). A cada três dias (D3, D6, D9 e D12), três leitões de cada grupo experimental foram eutanasiados e posteriormente submetidos à necropsia. Amostras de intestino (íleo, ceco, linfonodos mesentéricos e linfonodos ileocólicos), fígado, baço e pulmões foram coletadas para o isolamento de Salmonella. Os resultados mostram que, numericamente, o isolamento de Salmonella nos órgãos do G2 foi inferior ao G1, e que o número de amostras positivas de ceco no G1 (66,7%; 8/12) foi estatisticamente diferente do número de amostras positivas no G2 (16,7%; 2/12), com redução de 28,6% do total de amostras positivas de ceco no grupo tratado em relação ao controle. Portanto, observou-se que o ácido orgânico líquido foi capaz de reduzir a colonização de órgãos por Salmonella Typhimurium.(AU)

Identification and characterization of pathogenic and multidrug-resistant bacteria in feral pigeons surrounding a veterinary hospital in Minas Gerais, Brazil / Identificação de bactérias patogênicas e multirresistentes a antimicrobianos em pombos urbanos no entorno de um Hospital Veterinário em Belo Horizonte, Minas Gerais, Brasil

Pigeons are known for their capacity to harbor and spread several zoonotic agents. Studies have suggested that pigeons are also relevant disseminators of multidrug-resistant strains. In this study, pigeons surrounding a veterinary hospital were sampled and tested for the presence of pathogenic Escherichia coli, Salmonella spp., Staphylococcus spp., and Clostridioides (Clostridium) difficile. E. coli isolates from 19 (40.4%) pigeons tested positive for the E. coli heat-stable enterotoxin 1 (EAST1)-encoding gene. The intimin-encoding gene (eae) of enteropathogenicE. coli (EPEC) was found in one isolate (2.1%). Salmonella spp. were found in nine (19.1%) pigeons, all from the first capture event (P < 000.1). S. Typhimurium and S. Heidelberg were isolated from six and three pigeons, respectively. Enterobacterial repetitive intergenic consensus (ERIC-PCR) of the Salmonella spp. isolates suggested that eight of the nine strains had a high genetic similarity, supporting the hypothesis of an outbreak of salmonellosis in these pigeons. Twenty (42.5%) staphylococcal isolates were recovered from 18 (38.3%) pigeons. Eight different species were detected, with S. xylosus being the most frequent. Two (4.3%) C. difficile strains were isolated. Three isolates, one each of S. Typhimurium, S. aureus, and C. difficile, were classified as multidrug-resistant strains. The present research suggested that pigeons residing in urban areas can act as reservoirs and disseminators of pathogenic bacteria, including nosocomial pathogens, such as diarrheagenicE. coli and multidrug-resistant Staphylococcus spp., C. difficile, and Salmonella spp.

Pombos urbanos são conhecidos pela sua capacidade de carrear e disseminar diversos agentes zoonóticos. Estudos tem sugerido que pombos são também relevantes na disseminação de estirpes resistentes a múltiplas drogas. No presente estudo, pombos no ambiente de um hospital veterinário foram amostrados em três diferentes períodos e testados para a presença de Escherichia coli patogênica, Salmonella spp., Staphylococcus spp. e Clostridioides (Clostridium) difficile. Isolados de E. coli de 19 pombos (40.4%) foram positivos para o gene codificador da toxina EAST1. O gene codificador de intimina (eae) do patotipo E. coli enteropatogênica foi encontrada em um isolado (2.1%). Salmonella spp. foi encontrada em nove pombos (19.1%), sendo todos isolados do primeiro período de captura (P < 000.1). S. Typhimurium foi isolado de seis animais e S. Heidelberg de três. A tipagem molecular de isolados de Salmonella spp. por ERIC-PCR demonstrou que oito estirpes possuíam alta similaridade genética entre si, sugerindo a ocorrência de um surto de salmonelose nos animais carreadores. Vinte Staphylococcus (42.5%) foram isolados de 18 animais (38.3%). Oito diferentes espécies foram detectadas, sendo S. xylosus a mais frequente. Duas estirpes de C. difficile não-toxigênica (4.3%) foram isoladas. Uma estirpe de S. Typhimurium, uma de S. aureus e um isolado de C. difficile foram classificados como resistentes a múltiplas drogas antimicrobianas. O presente estudo sugere que pombos capturados no ambiente do hospital veterinário podem atuar como reservatórios e disseminadores de bactérias patogênicas e envolvidas em infecção hospitalar, incluindo E. coli diarreiogênica e Staphylococcus sp., C. difficile e Salmonella spp multirresistente.

Occurrence of Salmonella spp. in fecal samples from foals with and without diarrhea in the state of São Paulo: microbiological diagnosis, antimicrobial susceptibility profile, and molecular detection / Ocorrência de Salmonella spp. em amostras fecais de potros com e sem diarreia no estado de São Paulo: diagnóstico microbiológico, perfil de susceptibilidade aos antimicrobianos e detecção molecular

ABSTRACT: The present study investigated Salmonella spp. in the feces of 200 foals up to one year of age (100 with clinical signs of diarrhea and 100 without clinical signs of diarrhea). Bacteriological culture, serotyping, antimicrobial susceptibility, and real-time PCR (qPCR SYBR® Green or a TaqMan®) for detecting the invA gene (with and without a selective pre-enrichment step in tetrathionate broth) were performed. Bacterial culture revealed 15% (n=30) of positive animals (21 animals with diarrhea and nine without diarrhea). Among the 30 isolates, 13 different serovars were identified: S. Infantis, S. Minnesota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. Braenderup, S. Give, S. Newport, S. IIIb 61:c:z35, S. 109:-:1.5, S. I.4.12:d:-, S. I.6.8:-:-. Multidrug resistance was found in 43.33% (n=13) of the isolates, with one isolate obtained from animals without diarrhea and 12 isolates from animals with diarrhea. All qPCR techniques used in the study classified more samples as positive for Salmonella spp. than the bacterial culture of feces. In addition, all qPCR techniques detected more positive animals in the diarrhea group than in the diarrhea-free group. The results confirm the utility of the qPCR method without the pre-enrichment step in tetrathionate as a rapid test for Salmonella spp. in carrier animals. In animals with clinical signs of diarrhea, it can be combined with bacterial culture (antimicrobial susceptibility testing and serotyping). The isolation of Salmonella spp. in nine animals without diarrhea confirms the importance of asymptomatic carrier animals in the epidemiology of the disease. The multidrug resistance observed highlights the importance of rational antimicrobial use in horses and adopting biosecurity protocols that are efficacious in controlling the spread of infections between animals and zoonotic transmission in farms.

RESUMO: O presente estudo investigou a ocorrência de Salmonella spp. em fezes de 200 potros com até um ano de idade (100 com sinais clínicos de diarreia e 100 sem sinais clínicos de diarreia), utilizando as técnicas de cultivo bacteriológico e PCR em tempo real (qPCR) pelos métodos de corante fluorescente (SYBR® Green) e sonda específica (Taqman®) para a detecção do gene invA com e sem etapa de pré-enriquecimento seletivo em caldo de tetrationato. O cultivo bacteriológico revelou 15% (n=30) de animais positivos (21 animais com diarreia e nove animais sem diarreia). Dentre esses 30 isolados, 13 sorovares diferentes foram identificados: S. Infantis, S. Minnesota, S. I.4,5,12:i:-; S. Anatum, S. Cerro, S. Oranienburg, S. Braenderup, S. Give, S. Newport, S. IIIb 61:c:z35, S. 109:-:1.5, S. I.4.12:d:-, S. I.6.8:-:-. Multirresistência foi constatada em 43,33% (n=13) dos isolados, sendo um isolado obtido de animal sem diarreia e 12 isolados de animais com diarreia. Todas as técnicas de qPCR empregadas no estudo apresentaram maior número de amostras classificadas como positivas para Salmonella spp. comparadas ao cultivo bacteriológico de fezes. Adicionalmente, em todas as técnicas de qPCR houve maior número de animais detectados como positivos no grupo de animais com diarreia em relação aos animais sem diarreia. Os resultados confirmaram a utilidade do método qPCR sem a etapa de pré-enriquecimento em tetrationato, como um teste rápido para detecção de Salmonella spp. em animais portadores ou em animais com sinais clínicos de diarreia. O cultivo bacteriológico deve ser associado para a realização do teste de sensibilidade aos antimicrobianos e sorotipificação. O isolamento de Salmonella spp. em nove animais sem diarreia, confirma a importância dos animais portadores assintomáticos na epidemiologia da doença. A multirresistência observada evidencia a importância do uso racional de antimicrobianos em equinos e a importância da adoção de protocolos de biossegurança que sejam eficazes para controlar a disseminação de infecções entre animais e a transmissão zoonótica nas fazendas.

Autolyzed Saccharomyces cerevisae reduce microbiological carcass contamination in feedlot steers / Suplementação com Saccharomyces cerevisaeautolisada reduz contaminação microbiológica da carcaça de novilhos terminados em confinamento

The present work aims to verify whether the autolyzed Saccharomyces cerevisiae reduces total coliforms, Escherichia coli and Salmonella spp. from the hide, feces and carcass of feedlot-finished steers. Eighteen half-Angus steers were subjected to three treatments daily for 105 days: control (n=6), Lev 4 g (diet with 4 g yeast animal day-1, n=6) or Lev 7 g (diet with 7 g yeast animal day-1, n=6). On Days 29 and 90 after entrance into the feedlot, fecal samples, skin swabs, water and food were collected to identify and count E. coli and total coliforms and identify Salmonella spp. On the day of slaughter, carcasses were collected to identify and quantify E. coli, total coliforms, and mesophiles. There were reductions in E. coli and total coliforms in fecal samples (P=0.0001 and 0.001, respectively) and mesophilic aerobics in carcasses (P= 0.05) in the treated groups, and no Salmonella spp. was observed in any of the samples collected. It was concluded that supplementation reduced fecal excretion of and, consequently, carcass contamination by E. coli, mesophiles and total coliforms in confined animals and that this technique is a beneficial and residue-free sanitary measure to improve the microbiological quality of meat.

Este trabalho objetivou verificar se o prebiótico a base de parede celular de Saccharomyces cerevisiae reduz coliformes totais, Escherichia coli e Salmonella spp da pele, fezes e da carcaça de bovinos terminados em confinamento. Para tanto, 18 novilhos, ½ sangue Angus foram submetidos a três tratamentos diários durante 105 dias: controle (n=6), Lev 4g (dieta com 4 g de levedura por animal dia-1, n=6) ou Lev 7g ( dieta com 7 g de levedura por animal dia-1, n=6). Nos dias 29 e 90 após a entrada no confinamento, coletou-se amostras de fezes, swab de pele, água e alimento para contagem de Escherichia coli e coliformes totais e isolamento de Salmonella spp. No dia do abate, realizou-se coleta nas carcaças para identificação de Salmonella spp e quantificação de Escherichia coli, coliformes totais, mesófilos. Notou-se redução de Escherichia coli e coliformes totais nas amostras de fezes (P=0,0001 e 0,001 respectivamente) e de Escherichia coli, coliformes totais e aeróbios mesófilos na carcaça (P=0,06; 0,10 e 0,05 respectivamente) nos grupos tratados. Houve ausência de Salmonella spp. em todas as coletas realizadas. Concluiu-se que a suplementação reduziu a excreção fecal e consequentemente, a contaminação da carcaça pela Escherichia coli, mesófilos e coliformes totais dos animais confinados, sendo uma medida sanitária benéfica e livre de resíduos para melhorar a qualidade microbiológica da carne.

Drug resistance and susceptibility testing of Gram negative bacterial isolates from healthy cattle with different Lactam resistance Phenotypes from Shandong province China

Abstract The objective of this study was to evaluate the effectiveness of common antibiotics against different microorganisms in apparently healthy cattle in Shandong province and its suburb. A total of 220 nasal swab samples were collected and cultured for bacteriological evaluation. All the bacteria isolates after preliminary identification were subjected to antibiogram studies following disc diffusion method. It was found in the study that E. coli is the most commonly associated isolate (21%), followed by Klebsiella spp. (18%), Pseudomonas aeruginosa (13%), Salmonella spp. (15%), Shigella spp (12%), and Proteus spp (11%). While the antibiogram studies reveled that highest number of bacterial isolates showed resistance to Ampicillin (95%), followed by Augmentin (91%), Cefuroxime (85%) and Tetracycline (95%) of (Escherichia coli and Klebsiella spp). In the case of pseudomonas spp. and Salmonella the highest resistance was showed by Ampicillin (90%) followed by Amoxicillin + Clavulanic Acid (80%), Cefixime (90%), and Erythromycin (80%). In Shigella spp and Salmonella spp highest resistance was showed by Amoxicillin, Ceftazidime, Augmentin (60%), and Amoxicillin + Clavulanic Acid (50%). It is concluded that in vitro antibiogram studies of bacterial isolates revealed higher resistance for Ampicillin, Augmentin, Cefuroxime, Cefixime, Tetracycline, Erythromycin, and Amoxicillin + Clavulanic Acid. The high multiple Antibiotics resistance indexes (MARI) observed in all the isolates in this study ranging from 0.6 to 0.9. MARI value of >0.2 is suggests multiple antibiotic resistant bacteria and indicate presence of highly resistant bacteria.

Resumo O objetivo deste estudo foi avaliar a eficácia dos antibióticos comuns contra diferentes microrganismos em bovinos aparentemente saudáveis na província de Shandong e seus subúrbios. Um total de 220 amostras de esfregaço nasal foi coletado e cultivado para avaliação bacteriológica. Todos os isolados de bactérias após identificação preliminar foram submetidos a estudos de antibiograma seguindo o método de difusão em disco. Verificou-se no estudo que E. coli é o isolado mais comumente associado (21%), seguido por Klebsiella spp. (18%), Pseudomonas aeruginosa (13%), Salmonella spp. (15%), Shigella spp (12%) e Proteus spp (11%). Enquanto os estudos de antibiograma revelaram que o maior número de isolados bacterianos apresentou resistência à Ampicilina (95%), seguido por Augmentin (91%), Cefuroxima (85%) e Tetraciclina (95%) de (Escherichia coli e Klebsiella spp). No caso de Pseudomonas spp. e Salmonella, a maior resistência foi apresentada pela Ampicilina (90%) seguida pela Amoxicilina + Ácido Clavulânico (80%), Cefixima (90%) e Eritromicina (80%). Em Shigella spp e Salmonella spp, a maior resistência foi demonstrada por Amoxicilina, Ceftazidima, Augmentina (60%) e Amoxicilina + Ácido Clavulânico (50%). Conclui-se que estudos de antibiograma in vitro de isolados bacterianos revelaram maior resistência para Ampicilina, Augmentina, Cefuroxima, Cefixima, Tetraciclina, Eritromicina e Amoxicilina + Ácido Clavulânico. Os altos índices de resistência a antibióticos múltiplos (MARI) observados em todos os isolados neste estudo variaram de 0,6 a 0,9. O valor MARI de > 0,2 sugere várias bactérias resistentes a antibióticos e indica a presença de bactérias altamente resistentes.

Análise microbiológica de sushi comercializado em Rio Branco, Acre / Microbiological analysis of sushi marketed in Rio Branco, Acre / Análisis microbiológico de sushi comercializado en Rio Branco, Acre

O Brasil é um dos países mais diversificados no ramo gastronômico oferecendo vários alimentos diferentes aos seus consumidores, com base nos próprios pratos típicos ou provenientes de outras culturas. O pescado trata-se de um alimento perecível que necessita de atenções especiais em seu processamento. Falhas nas condições higiênico-sanitárias, associadas com a não cocção do alimento, podem ocasionar em uma contaminação e proliferação de bactérias, o que leva à uma grande preocupação a nível de saúde pública. O estudo analisou os aspectos microbiológicos de sushi comercializado na cidade de Rio Branco ­ Acre verificando os parâmetros de qualidade e as condições higiênicas sanitárias, comparando os resultados obtidos com a legislação vigente estabelecida pela ANVISA. Foram escolhidos 5 estabelecimentos aleatoriamente, sendo escolhidas 3 amostras de sushis do tipo niguiri de cada. As análises microbiológicas incluíram coliformes totais e coliformes termotolerantes utilizando a técnica dos tubos multiplos e a técnica de semeadura por profundidade para mesófilos e Salmonella. Constatou-se que todas as amostras tiveram um crescimento bacteriano e presença sugestiva de Salmonella, tornando o alimento impróprio para o consumo e mostrando uma falha nas condições higiênico- sanitária ao qual o sushi é processado e armazenado. É necessário maior fiscalização dos órgãos responsáveis e cuidado dos estabelecimentos que vendem sushi na cidade de Rio Branco, para que o produto vendido seja de boa qualidade e não cause malefícios a saúde de quem o consome.(AU)

Brazil is one of the most diversified countries in the gastronomic field, offering several different foods to its consumers, based on typical dishes or from other cultures. Fish is a perishable food that requires special attention in its processing. Failures in hygienic-sanitary conditions, coupled with the consumption of undercooked food, can lead to contamination and the proliferation of bacteria, which raises significant concerns regarding public health. The study analyzed the microbiological aspects of sushi sold in the city of Rio Branco - Acre, verifying the quality parameters and the hygienic sanitary conditions, comparing the obtained results with the current legislation established by ANVISA. Five establishments were randomly selected, and three samples of nigiri sushi were chosen from each establishment. The microbiological analysis included total coliforms and thermotolerant coliforms using the multiple tube technique, as well as depth seeding technique for mesophiles and Salmonella. It was found that all samples exhibited bacterial growth and suggested the presence of Salmonella, rendering the food unsuitable for consumption and indicating a failure in the hygienic-sanitary conditions under which the sushi was processed and stored. Greater inspection by the responsible authorities and improved care by establishments selling sushi in the city of Rio Branco are necessary to ensure that the product sold is of good quality and does not pose harm to the health of consumers.(AU)

Brasil es uno de los países más diversificados en el campo gastronómico, ofreciendo muchos alimentos diferentes a sus consumidores, basados en platos típicos ode otras culturas El pescado es un alimento perecedero que necesita especial atención en su elaboración. Las fallas en las condiciones, higiénico-sanitarias asociadas a la no cocción de los alimentos, pueden conducir a la contaminación y proliferación de bacterias, lo que genera una gran preocupación en términos de salud pública. El estudio analizó los aspectos microbiológicos del sushi comercializado en la ciudad de Rio Branco - Acre, verificando los parámetros de calidad y las condiciones higiénicas sanitarias, comparando los resultados obtenidos con la legislación vigente establecida por la ANVISA. Se eligieron 5 establecimientos al azar, y de cada uno se escogieron 3 muestras de sushi niguiri. Los análisis microbiológicos incluyeron coliformes totales y coliformes termotolerantes mediante la técnica de tubos múltiples y la técnica de siembra profunda para mesófilos y Salmonella. Se encontró que todas las muestras presentaban crecimiento bacteriano y la sugestiva presencia de Salmonella, lo que hace que el alimento no sea apto para el consumo y presenta una falla en las condiciones higiénico-sanitarias en las que se procesa y almacena el sushi. Se necesita mayor fiscalización por parte de los órganos responsables y cuidado de los establecimientos que venden sushi en la ciudad de Rio Branco, para que el producto vendido sea de buena calidad y no cause daño a la salud de quien lo consume.(AU)

Effect of Bacillus Subtilis on Immune Function of Hd11 Chicken Macrophages

Poultry is frequently contaminated by Salmonella, a pathogen leading to human health concern worldwide. This study aimed to evaluate the effect of Bacillus subtilis (BS)strain 048 (BS048) on the activation, phagocytosis, sterilization, cytokine secretion, and nitrogen oxide synthesis of HD11 chicken macrophages subjected to Salmonella enteritidis challenge, using lipopolysaccharide treatment as a negative control. The results showed: (1) BS048 had no significant effect on extracellular lactate dehydrogenase activity (p>0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p >0.05), while lipopolysaccharide treatment significantly increased extracellular lactate dehydrogenase activity (p 0.05);(2)BS048 significantly upregulated the expression levels of pro-inflammatory cytokines (interleukin (IL)- 1â and IL-6), anti-inflammatory cytokines(IL-10 and transforming growth factor-â1), and anti-viral cytokine, interferon-â (p<0.01); ; (3) BS048 significantly upregulated the mRNA expression level of the inducible nitric oxide synthase and its activity as well as extracellular nitrogen oxide level (p <0.01). In conclusion, BS048 could improve antiinflammatory and immune functions of HD11 chicken macrophages, without cytotoxic effects on these cells.(AU)

Avaliação da qualidade microbiológica de saladas de frutas comercializadas em Recife-PE / Evaluation of the microbiological quality of fruit salad commercialized in Recife-PE

Para auxiliar na mudança de vida e melhorar os hábitos alimentares, a população vem buscando integrar no seu cotidiano uma alimentação prática, saudável e nutritiva. Nesse contexto, a salada de frutas se torna um aliado por fornecer vitaminas e minerais importantes para a função básica do organismo, porém para tornar esse alimento seguro faz-se necessário um controle de sua qualidade. Por ser um alimento que contém um alto teor de água, sendo propício ao desenvolvimento de bactérias pode apresentar risco para a saúde do consumidor, caso não obtenham cuidados higiênicos-sanitários durante o seu processo produtivo. O presente estudo objetivou-se verificar a qualidade microbiológica das saladas de frutas prontas para consumo comercializadas na cidade do Recife-PE. Foram analisadas presença ou ausência de Salmonella sp. e a enumeração de coliformes termotolerantes a 45° C em vinte amostras do alimento coletado. A metodologia utilizada foi a descrita por Silva et al. (2017) e os resultados comparados com os parâmetros contidos na RDC nº 331 de 2019 e na Instrução Normativa nº 60 de 2019. Todas as amostras apresentaram ausência para Salmonella sp. em 25 g, mas para Coliformes termotolerantes foi detectada presença em 100% das amostras embora os valores estão dentro do que foi determinado na legislação. Conforme os resultados obtidos neste trabalho, as saladas de frutas estão aptas para consumo segundo as normas vigentes.(AU)

To help change their lives and improve eating habits, the population has been seeking to integrate into their daily lives a practical, healthy and nutritious diet. In this context, fruit salad becomes an ally for providing important vitamins and minerals for the basic function of the body, but to make this food safe it is necessary to control its quality. Because it is a food that presents a high water content, being prone to the development of bacteria and presenting a risk to the consumer's health, if hygienic-sanitary care is not taken during its production process. The present study aimed to verify the microbiological quality of ready-to-eat fruit salads sold in the city of Recife-PE. The presence or absence of Salmonella sp. and the enumeration of thermotolerant coliforms at 45°C in twenty samples of the food collected were analyzed. The methodology used was the one described by Silva et al. (2017) and the results were compared with the parameters contained in the RDC Nº 12 of 2001 and in the Normative Instruction Nº60 of 2019. All samples showed absence for Salmonella sp. in 25g, as for thermotolerant Coliforms was detected presence in 100% of products, but both values are within what was determined in the legislation. According to the results obtained in this study, the fruit salads are fit for consumption according to the current regulations.(AU)

Microbiological featuring of two neotropical fish species from the quilombola area of maranhão state, Brazil / Caracterização microbiológica de duas espécies de peixes neotropicais oriundas de área quilombola maranhense, Brasil

The aim of study is to feature the microbiological quality of two neotropical fish species from the quilombola area of Maranhão State, Brazil. In order to do so, 21 samples of Hoplerythrinus unitaeniatus and 21 samples of Cichlasoma bimaculatum were captured in flooded environment. Collected fish were euthanized in laboratory environment; muscle fragments were removed for microbiological analyses focused on enumerating molds and yeasts, viable strict and facultative mesophilic microorganisms and coagulase-positive staphylococci; on counting total and thermotolerant coliforms; and on investigating Escherichia coli and Salmonella sp. Microbiological results were compared to the Brazilian legislation, which establishes the list of microbiological standards for food products. Among the assessed fish, 9.52% were classified as non-acceptable for human consumption, based on the Salmonella parameter. Enumerated coagulase-positive staphylococci ranged from < 10 to 3.9 x 104 CFU/g; 9.52% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. E. coli counting ranged from 3.6 to > 1,100MPN/g; 4.76% of assessed fish were classified as having intermediate standard for human consumption, whereas 4.76% were classified as non-acceptable for such a purpose. Total and thermotolerant coliforms' counting and the enumeration of viable strict and facultative aerobic microorganisms, as well as of molds and yeasts, have evidenced high microbial population rates; this finding suggests poor hygienic conditions at capture site, contaminated raw material and risk of incidence of enteropathogens. This finding has evidenced imbalance in the investigated environment, as well as compromised aquatic biodiversity.(AU)

Objetivou-se caracterizar a qualidade microbiológica de duas espécies de peixes neotropicais oriundas de área qui-lombola maranhense, Brasil. Para isso, foram capturadas 21 amostras de Hoplerythrinus unitaeniatus e 21 amostras de Cichlasoma bimaculatum de ambiente alagável. No laboratório, os peixes foram eutanasiados, procedida a retirada dos fragmentos mus-culares e realizada as análises microbiológicas: enumeração de bolores e leveduras, micro-organismos mesófilos aeróbios estri-tos e facultativos viáveis e de estafilococos coagulase positiva, quantificação de coliformes totais e termotolerantes, pesquisa de Escherichia coli e Salmonella sp. Os resultados microbiológicos obtidos foram comparados com a legislação brasileira que estabelece a lista de padrões microbiológicos para alimentos. Dos peixes avaliados, 9,52% foram considerados inaceitáveis para consumo humano para o parâmetro Salmonella. A enumeração de estafilococos coagulase positivavariou de <10 a 3,9 x 104 UFC/g, sendo 9,52% dos peixes considerados com padrão intermediário e 4,76% inaceitáveis para consumo. A quanti-ficação de E. coli variou de 3,6 a >1.100 NMP/g, sendo 4,76% considerados com padrão intermediário e 4,76% inaceitáveis para consumo. A quantificação de coliformes totais e termotolerantes e a enumeração de micro-organismos aeróbios estritos e facultativos viáveis e bolores e leveduras revelou elevadas populações microbianas o que sugere más condições higiênicas do local de captura, matéria-prima contaminada e risco da presença de enteropatógenos, demonstrando desequilíbrio no ambiente estudado com comprometimento da biodiversidade aquática.(AU)

Portrayal of Punica granatum L. peel extract through High Performance Liquid Chromatography and antimicrobial activity evaluation

Abstract Increasing trend in antimicrobial resistance and failure of chemically synthesized antibiotics lead to discover alternative methods for the treatment of bacterial infections. Various medicinal plants are in use traditionally and their active compounds can be further applied for treatment of bacterial diseases. This study was designed to determine the antibacterial activity of Punica granatum (P. granatum L.) (pomegranate) peel extract against Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) and Shigella Dysenteriae (S. Dysenteriae)] and gram-positive bacterium [Staphylococcus aureus (Staph aureus)]. Methanolic extract of P. granatum L. peel was prepared by Soxhlet apparatus method. Total flavonoid and phenolic contents from the extract were determined by High Performance Liquid Chromatography (HPLC). The antibacterial activity of P. granatum L. peel extract was evaluated through agar well diffusion method. HPLC showed the range of phenolics (gallic acid, caffeic acid, benzoic acid, cinnamic acid) and flavonoid compounds. The chemical structures of flavonoid and phenolics found in the methanolic extract of P. granatum L. peel have been reported for the first time. The methanolic peel extract (50 ul) of yellow P. granatum L. showed 26, 10, 10 and 9mm zones of inhibition (ZOI) against S. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. The methanolic extract of red P. granatum L. (100 ul) showed 27, 8, 12 and 15 mm ZOI against Staph. aureus, S. Typhimurium, S. Dysenteriae and E. coli, respectively. Highest ZOI was observed against Staph. aureus. Many of the bacteria studied in the present work may cause serious gastrointestinal infections, which can lead to hemorrhagic diarrhea in children. These infections can be life-threatening to young children and the elderly. There is an incentive to find alternative control measures, such as plant and herbal extracts, especially in lesser-developed countries where traditional antibiotics may not be readily available.

Resumo A tendência crescente na resistência antimicrobiana e na falha dos antibióticos sintetizados quimicamente leva à descoberta de métodos alternativos para o tratamento de infecções bacterianas. Várias plantas medicinais estão em uso tradicionalmente e seus compostos ativos podem ser posteriormente aplicados para o tratamento de doenças bacterianas. Este estudo foi desenhado para determinar a atividade antibacteriana do extrato de casca de Punica granatum (P. granatum L.) (romã) contra Enterobacteriaceae [Escherichia coli (E. coli), Salmonella Typhimurium (S. Typhimurium) e Shigella Dysenteriae (S. Dysenteriae) ] e bactéria gram-positiva [Staphylococcus aureus (Staph aureus)]. O extrato metanólico da casca de P. granatum L. foi preparado pelo método do aparelho de Soxhlet. O conteúdo total de flavonoides e fenólicos do extrato foi determinado por cromatografia líquida de alta eficiência (HPLC). A atividade antibacteriana do extrato da casca de P. granatum L. foi avaliada através do método de difusão em ágar. HPLC mostrou a gama de compostos fenólicos (ácido gálico, ácido cafeico, ácido benzoico, ácido cinâmico) e flavonoides. As estruturas químicas de flavonoides e fenólicos encontradas no extrato metanólico da casca de P. granatum L. foram relatadas pela primeira vez. O extrato metanólico da casca (50 ul) de P. granatum L. amarelo apresentou zonas de inibição (ZOI) de 26, 10, 10 e 9mm contra S. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O extrato metanólico de P. granatum L. vermelho (100 ul) apresentou 27, 8, 12 e 15 mm IOI contra Staph. aureus, S. Typhimurium, S. Dysenteriae e E. coli, respectivamente. O ZOI mais alto foi observado contra Staph. aureus. Muitas das bactérias estudadas no presente trabalho podem causar infecções gastrointestinais graves, que podem levar à diarreia hemorrágica em crianças. Essas infecções podem ser fatais para crianças pequenas e idosos. Há um incentivo para encontrar medidas de controle alternativas, como extratos de plantas e ervas, especialmente em países menos desenvolvidos, onde os antibióticos tradicionais podem não estar prontamente disponíveis.