Efeito da vacina contra coccidiose sobre a colonização de Salmonella enteritidis em frangos de corte tratados com microbiota cecal anaeróbia

Avaliou-se o efeito da vacina contra coccidiose sobre a habilidade da microbiota cecal anaeróbia (MCA), administrada por diferentes vias (aves tratadas), em proteger pintos de corte da colonização por Salmonella enteritidis (Se). Utilizaram-se 120 aves assim divididas: grupo A, pulverização de MCA em aves vacinadas contra coccidiose e desafiadas com Se, grupo B, inoculação endoesofágica de MCA em aves vacinadas e desafiadas, grupo C, MCA na água de bebida de aves vacinadas e desafiadas, grupo D, aves não tratadas, vacinadas e desafiadas, grupo E, aves não tratadas, não vacinadas e desafiadas e, grupo F, aves não tratadas, não vacinadas e não desafiadas (controle negativo). Utilizaram-se como parâmetros a colonização do trato digestivo por Se e sua presença nas fezes, bem como o peso corporal das aves, avaliados aos dois, sete e 12 dias após o desafio. Nas aves tratadas com MCA, especialmente por meio de pulverização e inoculação endoesofágica, a colonização do ceco por Se e sua presença nas fezes foram menores, mostrando que a ação da MCA contra a colonização de ceco e excreção fecal não foi afetada pelo uso da vacina contra coccidiose em pintos de corte e que a associação MCA/vacina contra coccidiose pode ser utilizada sem que haja comprometimento na eficácia da MCA. Nos grupos que não receberam MCA, vacinados ou não contra coccidiose, houve aumento da colonização cecal, bem como excreção fecal da amostra desafio. O ceco foi o local de maior presença e persistência da Se. O resultado de administração de MCA pela água de bebida não foi tão eficiente, mas somente este tratamento resultou em peso corporal das aves significativamente superior ao das aves do grupo não tratado, não vacinado e desafiado, indicando que a presença de salmonelas paratifóides não interfere na produtividade de frangos de corte. Não se observou diferença no peso das aves dos grupos D (vacinados contra coccidiose) e E (não vacinadas) desafiadas com Se, demonstrando que a vacina não influenciou negativamente o ganho de peso das aves desafiadas com Salmonela enteriditis.

Avaliação in vitro da atividade inibitória de Lactobacillus spp., isolados do inglúvio e cecos de aves sobre Salmonella

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detection and transfer of antimicrobial resistance gene integron in Salmonella Enteritidis derived from avian material

The expansion of global poultry production has increased the need to reduce or control the agents responsible for economic losses, including Salmonella spp. These bacteria are also of public health concern due to their potential to cause food poisoning, and, more recently, due to the antimicrobial resistance presented by these bacteria. Molecular biology is an important tool currently used in the diagnosis and research studies of main poultry diseases. The present studied analyzed 100 samples of Salmonella Enteritidis (SE) isolated from avian material aiming at detecting the class 1 integron gene, Integroninvolved in antimicrobial resistance, by means of polymerase chain reaction (PCR), and comparing it with plate inhibition test. Subsequently, SE samples were evaluated for their capacity to horizontally transfer this gene. There was no direct relationship between the presence of the class 1 integron gene and SE resistance to the 14 antimicrobials tested, as 80% of the studied samples were resistant to up to three antimicrobials, and did not present the aforementioned gene. However, horizontal transfer of this gene was accomplished in vitro (from Escherichia coli to Salmonella Enteritidis), demonstrating that capacity class 1 integron gene can be disseminated among enterobacteria.

Transference in vitro of the resistance to the antimicrobials between Escherichia coli, Lactobacillus spp. and Salmonella enteritidis isolated from chickens

O objetivo deste trabalho foi verificar a possibilidade de transferência de resistência aos antimicrobianos entre bactérias normais da microbiota de frangos e Salmonella Enteritidis. Utilizamos amostras de Lactobacillus spp. (L. spp.), Salmonella Enteritidis (SE) e Escherichia coli (E. coli) previamente isolados de frangos, selecionados após prova de sensibilidade antimicrobiana in vitro conforme metodologia padrão (Comitê Nacional para Padrões Clínicos de Laboratório). Utilizamos aqueles com resistência e sensibilidade aos antimicrobianos indutores, chamados de bactérias doadoras e receptoras, respectivamente. Os antimicrobianos indutores foram utilizados para estimular a transferência de resistência aos antimicrobianos entre as bactérias. A possibilidade de transferência foi verificada da E. coli resistente para a SE e L. spp. Também foi verificada a transferência de uma amostra de L. spp resistente aos antimicrobianos indutores para a SE. Só foi possível verificar a transferência da resistência aos antimicrobianos indutores quando a bactéria doadora foi a E. coli e a bactéria receptora foi a SE. No presente estudo concluímos que a transferência de resistência aos antimicrobianos entre bactérias é possível, mas nem todas as bactérias participam desse evento, não transmitindo e nem adquirindo esta resistência.

Use of anaerobic cecal microflora, lactose and acetic acid for the protection of broiler chicks against experimental infection with Salmonella Typhimurium and Salmonella enteritidis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Use of three rapid detection systems to evaluate the prevalence and dissemination of Salmonella in a Brazilian poultry slaughterhouse

Prevalence and dissemination of Salmonella in a Brazilian poultry slaughterhouse were evaluated by three rapid detection systems (SS/SV(TM), VICAM, OSRT(TM), Unipath/Oxoid, and REVEAL(TM), Neogen), plus the conventional procedure. The carcasses were sampled after bleeding (P1), defeathering (P2), evisceration (P3), washing (P4), chilling (P5) and the packaged end-product (P6). In the first set of carcasses, the Salmonella incidence determined by the conventional method was 38.3% and 22.5% by SS/SV(TM). In the set for evaluation of OSRT(TM), the number of positive samples was the same detected by the cultural procedure (49.0%). In the third set, the positivity by the conventional procedure was 33.3%, and 5.0% by REVEAL(TM). The comparisons of positives in the first and third sets of carcasses were significantly different (P < 0.05). The positivity for Salmonella, in carcasses at P1 to P6, as determined by at least one of the methods, was 47.5%, 47.5%, 32.5%, 30.0%, 30.0% and 37.7%, respectively.

A refrigeração no diagnóstico de Salmonella spp. utilizando o método microbiológico tradicional e reação em cadeia da polimerase em carcaças de frango

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antibacterial effects of Brazilian and Bulgarian propolis and synergistic effects with antibiotics acting on the bacterial DNA and folic acid

Propolis is a honeybee product that has been used since ancient times because of its therapeutic effects. It can be used in the development of alternative therapies for the treatment of many diseases, and because propolis shows antibacterial action, this work was carried out in order to investigate a possible synergism between propolis and antibiotics acting on DNA (ciprofloxacin and norfloxacin) and on the metabolism (cotrimoxazole) against Salmonella Typhi. Propolis samples collected in Brazil and Bulgaria were compared in these assays, and the synergism was investigated by using 1/2 and 1/4 of the minimal inhibitory concentration for propolis and antibiotics, evaluating the number of viable cells according to the incubation time. Brazilian and Bulgarian propolis showed antibacterial activity, but no synergistic effects with the three tested antibiotics were seen. Previous works by our laboratory have revealed that propolis has synergistic effects with antibiotics, acting on the bacterial wall and ribosome, but it does not seem to interact with antibiotics acting on DNA or folic acid, and only a bacteriostatic action was seen in these assay conditions.

Efficiency of prebiotics and probiotics on the performance, yield, meat quality and presence of Salmonella spp in carcasses of free-range broiler chickens

Two trials were carried out in the present study. Trial I evaluated the performance, carcass yield and breast meat quality, whereas Trial II evaluated the efficacy of utilizing prebiotics + probiotics on the control of Salmonella spp incidence in the carcasses of free-range broilers. In Trial I, 688 one-day-old male chicks of the Naked Neck Label Rouge strain were used, distributed in a randomized block design arranged according to a 2 x 2 factorial: control diet or diet supplemented with probiotics and prebiotics; and two rearing systems (confined or with access to paddocks - 3m²/bird), using four replicates with 35 birds each. The birds were reared until 84 days of age following the recommendations of management and nutrition for free-range strains, and had access to paddocks after 35 days of age. Water and food were given inside the experimental poultry house. Birds fed probiotics and prebiotics in the diet and the confined birds showed better performance, carcass yield and meat quality compared to the birds of the other treatments. In Trial II, 128 one-day-old male chicks of the free-range Naked Neck Label Rouge strain were used. The birds were distributed into four treatments: NCC (non-challenged control), NCS (non-challenged supplemented), CC (challenged control) and CS (challenged supplemented). There were no significant effects of adding probiotics and prebiotics in the diet in regard to Salmonella enteritidis recovery from the carcasses.

Effects of Brazilian and Bulgarian propolis on bactericidal activity of macrophages against Salmonella Typhimurium

Propolis has been used in folk medicine since ancient times due to its, many biological properties: such as antimicrobial. antinflammatory, antioxidant, immunomodulatory activities, among others. 'Macrophages play an important role in the early phase of Salmonella infection. In this work, macrophages were prestimated with Brazilian or Bulgarian propolis and subsequently challenged with Salmonella Typhimurium at different macrophage/bacteria ratio. After 60 min of incubation. cells were harvested with Triton-X to lyse the macrophages. To assess the bactericidal activity. The number of colony-forming units (CFU) of S. typhimurium was determined by plating 0.1 mL in 'Mueller Hinton agar. After 24 h. CFU were counted. and the percentage of bactericidal activity was obtained. Propolis from Brazil and Bulgaria enhanced the bactericidal activity of macrophages, depending on its concentration. Brazilian propolis seemed to be more efficient than that from Bulgaria. because of their different chemical composition. In Bulgaria, bees collect the material mainly from the bud exudate of poplar tree, while in Brazil, Baccharis dracunculifolia DC. was shown to be the main propolis source. Our data also showed that the increased bactericidal activity of macrophages involved the participation of oxygen (H2O2) and nitrogen (NO) intermediate metabolites. (C) 2004 Elsevier B.V. All rights reserved.

Evaluation of three enrichment broths and five plating media for Salmonella detection in poultry

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Production of reactive oxygen (H2O2) and nitrogen (NO) intermediates and TNF-α in mice genetically selected for high (H) and low (L) antibody response and experimentally infected with Leptospira serovar pomona

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Genotoxicity assessment of water soluble fractions of biodiesel and its diesel blends using the Salmonella assay and the in vitro MicroFlow (R) kit (Litron) assay

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Study of Salmonella typhimurium mutagenicity assay of (E)-piplartine by the Ames test

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Penetration time of Salmonella Heidelberg through shells of white and brown commercial eggs

This study aimed at determining the minimum time required for the penetration of Salmonella Heidelberg inside the eggs after contact with contaminated material. Recently-collected brown and white eggs from laying hens between 45-50 weeks of age, reared in a commercial poultry house, were artificially contaminated by contact with wood shavings moistened with liquid inoculum of Salmonella Heidelberg in stationary-growth phase (10³-10(4) CFU g-1). According to type (white or brown), eggs were distributed into three different groups, with four replicates each: negative control group (no artificial contamination), positive control group (analyzed externally immediately after contamination and internally after the maximum storage period of the test group) and test group. Eggs were stored at controlled environmental temperature varying from 25ºC to 30ºC. In the test group, eggs contents (yolk and albumen) were pooled and analyzed after 1:00, 1:30, 2:00, 2:30, 3:00, 3:30, and 4:00 hours after contamination for the presence of Salmonella Heidelberg in 25g of this pool. The experimental unit consisted of five eggs in each test. The analysis protocol included pre-enrichment, selective enrichment, plating on selective agar, and biochemical and serological tests. The results obtained were submitted to logistic regression, which indicated that the presence of Salmonella Heidelberg was verified after 2:16 h and 2:44 h of contact with white and brown eggs, respectively.