Virulence Genes and Resistance Profile of Escherichia coli Isolated in Meat Meal Samples

Background: Feed is the main route of transmission of pathogenic microorganisms and is responsible for a large part of the cost of poultry production, so the inclusion of alternative foods in diets for monogastrics has been a constant. Among alternative foods most used in modern poultry farming are animal meal, however, when contaminated they constitute a route of transmission of several pathogenic agents, including Escherichia coli. In addition, there is a zoonotic potential, as poultry products are intended for human consumption. The objective of this research was to detect virulence genes, as well as to evaluate the resistance profile of Escherichia coli isolates from meat meal samples.  Materials, Methods & Results: A total of 40 Escherichia coli isolates were analyzed and the virulence genes surveyed iss, ompT, hlyF, iutA, and fimA identified by Polymerase Chain Reaction (PCR). The antimicrobial agents tested were: amoxycillin (30 μg), ceftiofur (30 μg), ciprofloxacin (5 μg), doxycycline (30 μg), florfenicol (30 μg), fosfomycin (200 μg), gentamicin (10 μg), norfloxacin (10 μg) and oxacillin (1 μg). It was possible to observe the occurrence of the iss resistance gene in 100% of Escherichia coli isolates, followed by hlyF (85%), fimA (75%), ompT (17.5%) and iutA (5%). Regarding the simultaneous detection for the genes, a greater association between the genes iss, hlyF and fimA (60%) was verified. All isolates showed resistance to oxacillin (100%), followed by doxycycline (25%), amoxicillin (22.5%), norfloxacin (17.5%), ceftiofur (15%), florfenicol (12.5%), fosfomycin (12.5%), ciprofloxacin (10%) and gentamycin (2.5%). In this study, a variation of the multiple antimicrobial resistance index (IRMA) was observed between 0.22 and 0.77. Discussion: The indiscriminate use of of antimicrobials as performance enhancers in production animals, may have contributed to the increase in antimicrobial resistance, with the occurrence of multiresistant Escherichia coli carrying virulence genes. Virulence genes present in Escherichia coli isolates are studied to understand the degree of influence they exert in the establishment of the disease, one of the most researched genes is the iss gene, involved in the processes that promote serum resistance. In this study, iss (100%) was present in all the isolates analyzed, although it is not the only mechanism used by these bacteria to reach internal organs and trigger an infection, this gene encodes an important mechanism associated with high levels of virulence. The second highest prevalence found was of the hlyF gene (85%), the high prevalence of hlyF suggests virulence potential, involved with the production of hemolysin and improvement of outer membrane vesicles associated with the release of toxins. The fimA gene (75%) was detected in a slightly lower percentage when compared to iss and hlyF. With the second lowest prevalence, the ompT gene (17.5%), is involved in a process that includes the proteolytic degradation of antimicrobial peptides and with the lowest prevalence the iutA gene (5%). Certain combinations of virulence genes make the strains easier to survive, adhere to, colonize and even the ability to develop septicemic conditions. Multiresistant E. coli strains, is a fact of concern for both animal and human health, since the presence of multiresistant strains, originating from poultry, can be transmitted from chicken carcasses.  In this sense, the importance of sanitary control of the inputs used in animal feed is emphasized, as well as the prudent use of antimicrobials in animal production, with a view to producing a safe food, minimizing not only the economic losses, but also the risks to human health. Keywords: antimicrobial, bacterial resistance, colibacillosis, poultry.

CARACTERIZAÇÃO CLÍNICA DE MULHERES SUBMETIDAS À HISTERECTOMIA OBSTÉTRICA POR CONSEQUÊNCIA DA HEMORRAGIA PÓS-PARTO

Objetivo: Descrever as principais características clínica de mulheres submetidas à histerectomia obstétrica por consequência da hemorragia pós-parto. Métodos: estudo descritivo-exploratório, de abordagem quantitativa e caráter documental. Adotou-se como critérios de inclusão, prontuários de mulheres que pariram via parto vaginal ou cesariana, com idade igual ou acima de 18 anos; que tenham sido submetidas a histerectomia obstétrica em decorrência da hemorragia pós-parto; e submetidas ao procedimento cirúrgico de janeiro de 2018 a junho de 2022. Diante disso, 15 prontuários atenderam aos critérios. Resultados: a cesariana foi a principal via de parto (93,3%) e o sofrimento fetal a maior indicação dessa via (21,4%). A hemorragia pós-parto foi classificada como primária (93,3%) e teve a atonia uterina como maior causa (86,6%). A histerectomia subtotal foi a técnica cirúrgica mais utilizada (93,3%) e o tempo médio da expulsão do feto até a realização da histerectomia obstétrica ficou em 6,4 horas. Conclusão: Este estudo permitiu evidenciar a importância do tema tratado, uma vez que a hemorragia pós-parto é uma das principais causas de mortalidade materna em todo mundo. Portanto, conhecer o perfil clínico dessas mulheres é primordial para fornecer assistência de qualidade além de ofertar subsídios para protocolos e pesquisas futuras

A NOVA NATUREZA JURÍDICA DA ÁGUA E SUAS CONSEQUÊNCIAS EM FACE DA OUTORGA DE DIREITO DE USO DE RECURSOS HÍDRICOS.

O presente artigo busca elencar os novos aspectos legais referentes aos Recursos Hídricos definindo a água como bem de uso comum do povo, sendo essencial para a existência dos seres vivos. Tendo  como objetivo analisar os impactos e consequências práticas da nova dimensão jurídica que a Constituição da República Federativa do Brasil de 1988 (CRFB/88) trouxe ao meio ambiente no que tange ao elemento vital água.  Traz o entendimento do que seja um bem difuso e a noção de transindividualidade do Recurso Hídrico, conforme preceitua a CRFB/88. Aborda o recurso natural água que além de ser um bem difuso não integra o patrimônio do Poder Público, haja vista que ele é apenas seu gestor. Também abrange a outorga que é uma ferramenta desenvolvida pela Política Nacional dos Recursos Hídricos de extrema importância, uma vez que garante a real natureza de bem transindividual dos recursos hídricos. Para a elaboração deste artigo procedeu-se à pesquisa qualitativa mediante revisão bibliográfica

ATLANTIC EPIPHYTES: a data set of vascular and non-vascular epiphyte plants and lichens from the Atlantic Forest

Epiphytes are hyper-diverse and one of the frequently undervalued life forms in plant surveys and biodiversity inventories. Epiphytes of the Atlantic Forest, one of the most endangered ecosystems in the world, have high endemism and radiated recently in the Pliocene. We aimed to (1) compile an extensive Atlantic Forest data set on vascular, non-vascular plants (including hemiepiphytes), and lichen epiphyte species occurrence and abundance; (2) describe the epiphyte distribution in the Atlantic Forest, in order to indicate future sampling efforts. Our work presents the first epiphyte data set with information on abundance and occurrence of epiphyte phorophyte species. All data compiled here come from three main sources provided by the authors: published sources (comprising peer-reviewed articles, books, and theses), unpublished data, and herbarium data. We compiled a data set composed of 2,095 species, from 89,270 holo/hemiepiphyte records, in the Atlantic Forest of Brazil, Argentina, Paraguay, and Uruguay, recorded from 1824 to early 2018. Most of the records were from qualitative data (occurrence only, 88%), well distributed throughout the Atlantic Forest. For quantitative records, the most common sampling method was individual trees (71%), followed by plot sampling (19%), and transect sampling (10%). Angiosperms (81%) were the most frequently registered group, and Bromeliaceae and Orchidaceae were the families with the greatest number of records (27,272 and 21,945, respectively). Ferns and Lycophytes presented fewer records than Angiosperms, and Polypodiaceae were the most recorded family, and more concentrated in the Southern and Southeastern regions. Data on non-vascular plants and lichens were scarce, with a few disjunct records concentrated in the Northeastern region of the Atlantic Forest. For all non-vascular plant records, Lejeuneaceae, a family of liverworts, was the most recorded family. We hope that our effort to organize scattered epiphyte data help advance the knowledge of epiphyte ecology, as well as our understanding of macroecological and biogeographical patterns in the Atlantic Forest. No copyright restrictions are associated with the data set. Please cite this Ecology Data Paper if the data are used in publication and teaching events. © 2019 The Authors. Ecology © 2019 The Ecological Society of Americ

Epidemiologia molecular das infecções por Mycoplasma ssp., Escherichia coli e Staphylococcus spp., em frangos de corte e poedeiras comerciais no estado de Pernambuco

Objetivou-se com este trabalho estudar a epidemiologia molecular das infecções por Mycoplasma spp., Escherichia coli e Staphylococcus spp., em frangos de corte e poedeiras comerciais no Estado de Pernambuco. Foram utilizadas 120 aves provenientes de 24 granjas, das quais 55 frangos de corte sadios, 35 com sinais respiratórios e 30 poedeiras comerciais com sinais respiratórios e digestivos. Para a pesquisa de Mycoplasma spp., utilizou-se o exame bacteriológico e a reação em cadeia da polimerase (PCR) e Nested-PCR. Para o isolamento de Escherichia coli foi utilizado o meio ágar Eosina Azul de Metileno (EMB), o teste de patogenicidade in vitro foi realizado em ágar oxalato de magnésio acrescido de vermelho congo, enquanto a PCR foi realizada para avaliar a presença de genes de virulência e determinação filogenética dos grupos em A, B1, B2 e D. Para o estudo de citotoxicidade, as amostras foram inoculadas em células Vero e para a detecção de plasmídios foi utilizada a extração de DNA e visualização do perfil plasmidial. Para o isolamento de Staphylococcus spp., utilizou-se o exame bacteriológico, com provas bioquímicas para a identificação das espécies. Verificou-se a formação de biofilme em ágar Vermelho Congo (ACR), e detecção do gene mecA pela PCR. Para Escherichia coli e Staphylococcus spp., realizou-se perfil de resistência através dos testes de disco difusão e Concentração Inibitória Mínima (CIM). Na PCR para Mycoplasma spp., observou-se que sete (29,17%) amostras foram positivas para MS e uma (4,17%) para MG em amostras de aves com sinais respiratórios, sendo a amostra positiva confirmada como cepa vacinal MG-F; no exame bacteriológico todas as amostras foram negativas. Dos 35 isolados de Escherichia coli submetidos a PCR, o número de isolados positivos para os genes de virulência foi: iss (16), iutA (10), hlyF (17), ironN (11), ompT (16), crl (10), fimA (14), tsh (5), papA (2), csgA (0) e iucA (2). Foram classificados filogeneticamente nos grupos A (71,42%) e B1 (28,58%). A presença de hemólise em ágar sangue foi superior à detecção do gene hlyF pela PCR. O teste de patogenicidade em ágar vermelho congo revelou cinco (14,29%) isolados positivos e para avaliação da citotoxicidade in vitro em células Vero todos os isolados foram negativos. No perfil de resistência aos antibiomicrobianos observou-se que 33 (94,28%) isolados foram resistentes a três ou mais antibióticos e a lincomicina apresentou o maior percentual de resistência (100%). Na CIM observou-se multirresistência a vários antimicrobianos. Quanto aos plasmídios observou-se frequência de 80,0% (28/35) nos isolados de E. coli, onde 16 isolados apresentaram plasmídio de 88 MDa. Das 24 amostras processadas foram isolados 16 Staphylococcus, sendo cinco coagulase-positiva (SCP) e 11 coagulase negativa (SCN), e no teste de produção de biofilme, seis (37,5%) isolados foram positivos. Na PCR para detecção do gene mecA todos os isolados foram negativos. Observou-se que 15 isolados apresentaram perfil de multirresistência a antimicrobianos. Com base nos resultados obtidos, essas bactérias participam da doença respiratória na população estudada e devem ser consideradas nas medidas de controle e tratamento nos plantéis avícolas em conjunto com a realização de testes in vitro.This investigation had the objective of studying the molecular epidemiology of infections by Mycoplasma spp., Escherichia coli and Staphylococcus spp., in broilers and commercial layers of the state of Pernambuco. A hundred and twenty birds were used from 24 flock of which 55 healthy broilers, 35 broilers and 30 commercial layers with respiratory and digestive signs. For the study of Mycoplasma spp., the bacteriological exam, the Polymerase Chain Reaction (PCR) and Nested-PCR were used. For the isolation of Escherichia coli the medium Eosine Methylene Blue Agar (EMB) was used. The pathogenicity test in vitro was carried out in Congo Red magnesium oxalate agar while the PCR was used, to evaluate the presence of virulence genes and the phylogenetic determination of groups A, B1, B2 and D. For the study of citotoxicity, the samples were inoculated in Vero cells. For the detection of plasmids, the extraction of DNA and the visualization of the plasmid profile was used. For Staphylococcus spp., isolation the bacteriological exam was used, with biochemical proof of species identification. The formation of the biofilm Congo Red Agar (ACR), and the detection of the mecA gene by PCR were verified. For Escherichia coli and Staphylococcus spp., a resistance profile was carried out through tests of disc diffusion and Minimum Inhibitory Concentration (MIC). In the PCR for Mycoplasma spp., it was observed that seven (29.17%) samples were positive for MS and one (4.17%) for MG, in samples of birds with respiratory signs, the positive sample was further confirmed as vacinal strain MG-F; in the bacteriological exam all samples were negative. From the 35 isolated Escherichia coli submitted to PCR, the number of positive E coli isolates for virulence genes of were: iss (16), iutA (10), hlyF (17), ironN (11), ompT (16), crl (10), fimA (14), tsh (5), papA (2), csgA (0) and iucA (2). They were phylogenetically classified into groups A (71.42%) and B1 (28.58%). The presence of hemolysis in agar blood was superior to the detection of the hlyF gene by the PCR. The pathogenicity test in Congo Red agar showed five (14.29%) positive isolated E coli, and for the cytotoxicity evaluation in vitro, in Vero cells, all the isolated E coli were negative. On the resistance profile to antibiomicrobials, it was observed that 33 (94.28%) of the E coli isolates were resistant to three or more antibiotics and that lincomicine presented the highest percentage of resistance (100%). On the CIM, multiresistance to various antimircrobials was observed. With respect to the plasmids, a frequency of 80.0% (28/35) was observed in E. coli isolates, where 16 E. coli isolates presented plasmid of 88 MDa. From the 24 processed samples 16 Staphylococcus isolates were, five being coagulasis-positive (SCP) and 11 coagulasis-negative (SCN). The biofilm production test, six (37.5%) isolates were positive. Regarding PCR for the detection of the mecA gene, all the isolates were negative. It was observed that 15 isolates presented a multiresistance profile to antimicrobials. Based on the results, these bacteria participate in the respiratory disease of the studied chickens populations and they must be considered in the control and treatment measures used in aviculture, together with the performance of in vitro tests.Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNP

Identificação e avaliação in vitro da atividade inibitória de Lactobacillus spp isolados do ingúvio e cecos de aves (Gallus gallus, Linneaus, 1758), em relação a sorotipos de Salmonella

A colonização da mucosa intestinal por grande e diversificado número de bactérias é achado normal no homem e nos animais, incluindo as aves. Produtos de exclusão competitiva e/ou probióticos quando administrados às aves, devem promover a redução na colonização de patógenos potencialmente perigosos à saúde avícola e humana, como bactérias do gênero Salmonella e conseqüente equilíbrio na microbiota intestinal normal. O presente trabalho teve por objetivos verificar a presença de Lactobacillus spp. na microbiota do inglúvio e cecos de aves, identificar as espécies através dos métodos PCR e bioquímico e avaliar a capacidade da atividade inibitória in vitro das espécies isoladas frente a oito sorotipos de Salmonella. Utilizou-se aves reprodutoras e de postura comercial como doadoras de inglúvio e cecos para o isolamento e identificação de Lactobacillus. As amostras isoladas foram caracterizadas como pertencentes ao gênero Lactobacillus através dos testes de coloração de Gram positiva, catalase negativa, produção de gás em glicose e H2S em Tríplice Sugar Iron (TSI) negativo. Posteriormente, submetidas ao padrão de fermentação dos carboidratos e a PCR, para identificação das espécies de Lactobacillus. O método da PCR apresentou-se mais sensível mostrando resultados diferentes do método bioquímico. No bioquímico identificou-se L. acidophilus, L. fermentum, L. reuteri, L. salivarius e L. sp., enquanto na PCR, L. reuteri, L. salivarius e L. sp. Todas as espécies de Lactobacillus isoladas do inglúvio e cecos de aves apresentaram amostras que inibiram Salmonella Enteritidis fagotipo 4, S. Enteritidis fagotipo 28, S.Typhimurium, S. Pullorum, S. Agona, S. Anatum, S. Dublin e S. Senftenberg. Palavras chaves: Identificação, Lactobacillus, PCR, inibição, Salmonella, aves.Intestinal mucosa colonization by a diverse and great number of bacteria is considered normal among men and animals including poultry. Competitive exclusion products and \or probiotics when given to poultry should promote pathogen colonization reduction which is potentially danger to human and poultry health like Salmonella and consequently a balance on normal intestinal microbiota The present work aimed at verifying Lactobacillus spp on crop microbiota and poultry cecum, identifying species through PCR and biochemical methods and to evaluate in vitro inhibitory activity ability from species isolated against eight Salmonella serotypes Production delivery and reproduction poultry were used as crop and cecum donors to isolate and identify Lactobacillus. Isolated samples were characterized as belonging to Lactobacillus species through coloration gram positive tests, negative catalase, glucose gas production and H2S in negative triple sugar iron (TSI). Afterwards, samples were subjected to carbohydrate fermentation pattern and to PCR to identify Lactobacillus species. PCR method has shown to be more sensitive presenting different results of biochemical methods. On biochemical method one identified: L. acidophilus, L. fermentum, L. reuteri, L. salivarius, and L. sp, while in PCR, L. reuteri, L. salivarius and L.sp. All Lactobacillus species isolated from poultry crop and cecum showed samples which inhibited Salmonella Enteritidis fagotype 4, S. Enteritidis fagotype 28, S.Typhimurium, S. Pullorum, S. Agona, S. Anatum, S. Dublin and S. Senftenberg.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Virulence Genes and Resistance Profile of Escherichia coli Isolated in Meat Meal Samples

Background: Feed is the main route of transmission of pathogenic microorganisms and is responsible for a large part of the cost of poultry production, so the inclusion of alternative foods in diets for monogastrics has been a constant. Among alternative foods most used in modern poultry farming are animal meal, however, when contaminated they constitute a route of transmission of several pathogenic agents, including Escherichia coli. In addition, there is a zoonotic potential, as poultry products are intended for human consumption. The objective of this research was to detect virulence genes, as well as to evaluate the resistance profile of Escherichia coli isolates from meat meal samples.  Materials, Methods & Results: A total of 40 Escherichia coli isolates were analyzed and the virulence genes surveyed iss, ompT, hlyF, iutA, and fimA identified by Polymerase Chain Reaction (PCR). The antimicrobial agents tested were: amoxycillin (30 μg), ceftiofur (30 μg), ciprofloxacin (5 μg), doxycycline (30 μg), florfenicol (30 μg), fosfomycin (200 μg), gentamicin (10 μg), norfloxacin (10 μg) and oxacillin (1 μg). It was possible to observe the occurrence of the iss resistance gene in 100% of Escherichia coli isolates, followed by hlyF (85%), fimA (75%), ompT (17.5%) and iutA (5%). Regarding the simultaneous detection for the genes, a greater association between the genes iss, hlyF and fimA (60%) was verified. All isolates showed resistance to oxacillin (100%), followed by doxycycline (25%), amoxicillin (22.5%), norfloxacin (17.5%), ceftiofur (15%), florfenicol (12.5%), fosfomycin (12.5%), ciprofloxacin (10%) and gentamycin (2.5%). In this study, a variation of the multiple antimicrobial resistance index (IRMA) was observed between 0.22 and 0.77. Discussion: The indiscriminate use of of antimicrobials as performance enhancers in production animals, may have contributed to the increase in antimicrobial resistance, with the occurrence of multiresistant Escherichia coli carrying virulence genes. Virulence genes present in Escherichia coli isolates are studied to understand the degree of influence they exert in the establishment of the disease, one of the most researched genes is the iss gene, involved in the processes that promote serum resistance. In this study, iss (100%) was present in all the isolates analyzed, although it is not the only mechanism used by these bacteria to reach internal organs and trigger an infection, this gene encodes an important mechanism associated with high levels of virulence. The second highest prevalence found was of the hlyF gene (85%), the high prevalence of hlyF suggests virulence potential, involved with the production of hemolysin and improvement of outer membrane vesicles associated with the release of toxins. The fimA gene (75%) was detected in a slightly lower percentage when compared to iss and hlyF. With the second lowest prevalence, the ompT gene (17.5%), is involved in a process that includes the proteolytic degradation of antimicrobial peptides and with the lowest prevalence the iutA gene (5%). Certain combinations of virulence genes make the strains easier to survive, adhere to, colonize and even the ability to develop septicemic conditions. Multiresistant E. coli strains, is a fact of concern for both animal and human health, since the presence of multiresistant strains, originating from poultry, can be transmitted from chicken carcasses.  In this sense, the importance of sanitary control of the inputs used in animal feed is emphasized, as well as the prudent use of antimicrobials in animal production, with a view to producing a safe food, minimizing not only the economic losses, but also the risks to human health. Keywords: antimicrobial, bacterial resistance, colibacillosis, poultry.

Zinc, manganese and copper amino acid complexed in laying hens' diets affect performance, blood parameters and reproductive organs development.

In the intestinal lumen, excess of oxides and sulfates interfere with the absorption of minerals due to competition from the same absorption site. Amino acids-mineral complexed (AACM) is intended to minimize these problems, which might be absorbed by different absorption sites. Then, a study including Zinc (Zn), Manganese (Mn) and Copper (Cu) from different sources was carried out to evaluate the performance, blood parameters and reproductive organs development of Brown Laying Hens. A total of 800 Lohmann Brown Lite were fed, from one-day-old to 182-days-old, Zn, Mn and Cu from different sources. Measurements were made from 105 to 182-days-old. The laying hens were distributed according to a completely randomized design with 20 replicates and 20 birds per experimental unit. The treatments consisted of a diet supplemented with 70, 70 and 8 mg/kg of Zn, Mn and Cu; respectively, from inorganic sources (IM). The second treatment contained 40, 40 and 2.75 mg/kg of Zn, Mn and Cu, respectively from IM plus 30, 30 and 5.25 mg/kg of Zn, Mn and Cu; respectively, from AACM sources. Performance and reproductive organs development (oviduct and ovary weight), tibia weight, liver weight, egg output and body weight, and blood variables were evaluated. Data were compared by Student's t-test (P < 0.05). Laying hens fed AACM reached 35% of egg output two days earlier and presented heavier tibia bone than the IM group. Those hens also presented greater oviduct weight, greater hematocrit and greater serum concentration of total leukocytes, erythrocytes, eosinophils, monocytes and the hormones T4 and FSH, than the hens fed IM. The supplementation of AACM in laying hens' diets since one-day-old improves the productive performance from the beginning of egg output to peak production, which is justified by better development of bones and oviduct, hormone production and immune system support