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AIM: To evaluate the use of organic acids (OAs) and competitive exclusion (CE) product administered continuously in the feed and transiently in drinking water on the control of Salmonella enterica subspecie enterica serotype Enteritidis (SE) prior to slaughter. METHODS AND RESULTS: The influence of treatments were evaluated on pH, population of the lactic acid bacteria (LAB) and bacteria of the family Enterobacteriaceae, concentration of volatile fatty acids and SE colonization in the crop and caecum. The birds were challenged with SE 24 h before being slaughtered, and then, the caeca and crop were removed and subjected to SE counts. Continuous administration of OAs reduced the population of bacteria from the Enterobacteriaceae family in both crop and caecum, positively influenced the butyric acid concentration and reduced SE colonization in the caecum. The diet supplemented with CE product positively influenced the quantity of LAB in the crop and caecum, elevated the butyric acid concentration and reduced both Enterobacteriaceae quantity and SE colonization in the caecum. There was no effect from administering the treatments via drinking water on the variables measured. CONCLUSIONS: Continuous supplementation in feed with OAs and CE product reduced SE colonization of the caeca. SIGNIFICANCE AND IMPACT OF THE STUDY: Supplementation of OAs and CE product in diet to turkeys can reduce the SE load, potentially leading to a lower contamination risk of meat during slaughter.

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Foregut fermentation occurs in mammalian ruminants and in one bird, the South American folivorous hoatzin. This bird has an enlarged crop with a function analogous to the rumen, where foregut microbes degrade the otherwise indigestible plant matter, providing energy to the host from foregut fermentation, in addition to the fermentation that occurs in their hindguts (cecum/colon). As foregut fermentation represents an evolutionary convergence between hoatzins and ruminants, our aim was to compare the community structure of foregut and hindgut bacterial communities in the cow and hoatzin to evaluate the influences of host phylogeny and organ function in shaping the gut microbiome. The approach used was to hybridize amplified bacterial ribosomal RNA genes onto a high-density microarray (PhyloChip). The results show that the microbial communities cluster primarily by functional environment (foreguts cluster separately from hindguts) and then by host. Bacterial community diversity was higher in the cow than in the hoatzin. Overall, compared with hindguts, foreguts have higher proportions of Bacteroidetes and Spirochaetes, and lower proportions of Firmicutes and Proteobacteria. The main host differences in gut bacterial composition include a higher representation of Spirochaetes, Synergistetes and Verrucomicrobia in the cow. Despite the significant differences in host phylogeny, body size, physiology and diet, the function seems to shape the microbial communities involved in fermentation. Regardless of the independent origin of foregut fermentation in birds and mammals, organ function has led to convergence of the microbial community structure in phylogenetically distant hosts.

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Inglúvios e cecos de reprodutoras comerciais de frangos de corte foram utilizados para o isolamento de Lactobacillus spp. As estirpes isoladas foram caracterizadas como Gram-positivo, catalase negativo, produtoras de gás em glicose, não produtoras de H2S em triple sugar iron e identificadas pela reação em cadeia da polimerase como Lactobacillus reuteri e Lactobacillus salivarius. A utilização da técnica spot-on-the-lawn para avaliação da inibição in vitro permitiu a análise de vários microrganismos simultaneamente. Todas as estirpes isoladas inibiram in vitro S. Enteritidis fagotipo 4, S. Enteritidis fagotipo 28, S. Typhimurium, S. Pullorum, S. Agona, S. Anatum, S. Dublin e S. Senftenberg.(AU)

Crops and ceca of commercial broiler breeders were used for the isolation of Lactobacillus spp. The isolated samples were characterized by Gram positive staining, negative catalase test, production of gas from glucose, negative for H2S production from triple sugar iron, and were identified by the polymerase chain reaction as Lactobacillus reuteri and Lactobacillus salivarius. The use of the spot-on-the-lawn technique, with modifications, for the evaluation of the in vitro inhibition made it possible the simultaneous analysis of several microrganisms. All the crop and ceca isolated microorganisms presented an in vitro inhibitory effect against strains of Salmonella Enteritidis fagotype 4, S. Enteritidis fagotype 28, S. Typhimurium, S. Pullorum, S. Agona, S. Anatum, S. Dublin, and S. Senftenberg.(AU)

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The purpose of this study was to characterize Candida isolates from crop of parrots. Forty baby parrots of genus Amazona, species aestiva and amazonica that were apprehended from wild animal traffic were used: 18 presented ingluvitis and 22 other alterations, but showing general debilitation. Samples were seeded on Sabouraud dextrose agar with chloramphenicol after be obtained by the introduction of urethral probe through the esophagus. Based on morphology and biochemical reactions (API 20C) Candida was confirmed; it was still searched the production of proteinase and phospholipase, virulence factors for Candida species. Candida spp. were isolated from 57.5 percent parrots, being 72.2 percent from birds with ingluvitis and 45.5 percent from without ones. Twenty-five strains of Candida were isolated, 60 percent and 40 percent, respectively from parrots with and without ingluvitis, and were speciated: 28 percent C. humicola, 24 percent C. parapsilosis, 20 percent C. guilliermondii, 20 percent C. famata, and 8 percent C. albicans. These results demonstrate that C. albicans is not the most frequent species isolated, and it is the first report that shows C. guilliermondii, C. famata, and C. humicola causing infection in parrots. Many isolates presented filamentation (76 percent), 100 percent produced proteinase and 68 percent phospholipase. The observation of Candida spp. producing virulence factors reinforce the pathogenic role of these yeasts in the cases studied.

O objetivo do presente trabalho foi caracterizar cepas de Candida spp. isoladas de inglúvio de papagaios. Foram utilizados 40 papagaios do gênero Amazona, espécies aestiva e amazonica, apreendidos de tráfico de animais selvagens: 18 apresentavam ingluvite e 22 outras alterações, mas todos mostrando sinais de debilitação geral. Colheram-se as amostras clínicas através da introdução de sonda uretral no esôfago dos animais e estas foram semeadas em ágar Sabouraud dextrose acrescido de cloranfenicol. A identificação das espécies de Candida foi baseada em características macro e micromorfológicas e comportamento bioquímico no kit API 20C; pesquisou-se ainda a produção das enzimas proteinase e fosfolipase. Isolou-se Candida spp. de 57,5 por cento dos papagaios, sendo 72,2 por cento de aves com ingluvite e 45,5 por cento de aves com outras afecções. Isolaram-se 25 cepas de Candida, 60 por cento e 40 por cento respectivamente de animais com e sem ingluvite. As espécies isoladas foram: C. humicola (28 por cento), C. parapsilosis (24 por cento), C. guilliermondii (20 por cento), C. famata (20 por cento) e C. albicans (8 por cento). Os resultados obtidos demonstram que C. albicans não é a espécie mais freqüentemente isolada e é a primeira vez que se relata C. guilliermondii, C. famata e C. humicola causando infecção em papagaios. A maioria dos isolados apresentou filamentação (76 por cento) e produziu a enzima fosfolipase (68 por cento) e todas as cepas produziram proteinase. O encontro de Candida spp., produzindo seus fatores de virulência, reforça o papel patogênico dessas leveduras nos casos estudados.

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The purpose of this study was to characterize Candida isolates from crop of parrots. Forty baby parrots of genus Amazona, species aestiva and amazonica that were apprehended from wild animal traffic were used: 18 presented ingluvitis and 22 other alterations, but showing general debilitation. Samples were seeded on Sabouraud dextrose agar with chloramphenicol after be obtained by the introduction of urethral probe through the esophagus. Based on morphology and biochemical reactions (API 20C) Candida was confirmed; it was still searched the production of proteinase and phospholipase, virulence factors for Candida species. Candida spp. were isolated from 57.5 percent parrots, being 72.2 percent from birds with ingluvitis and 45.5 percent from without ones. Twenty-five strains of Candida were isolated, 60 percent and 40 percent, respectively from parrots with and without ingluvitis, and were speciated: 28 percent C. humicola, 24 percent C. parapsilosis, 20 percent C. guilliermondii, 20 percent C. famata, and 8 percent C. albicans. These results demonstrate that C. albicans is not the most frequent species isolated, and it is the first report that shows C. guilliermondii, C. famata, and C. humicola causing infection in parrots. Many isolates presented filamentation (76 percent), 100 percent produced proteinase and 68 percent phospholipase. The observation of Candida spp. producing virulence factors reinforce the pathogenic role of these yeasts in the cases studied.(AU)

O objetivo do presente trabalho foi caracterizar cepas de Candida spp. isoladas de inglúvio de papagaios. Foram utilizados 40 papagaios do gênero Amazona, espécies aestiva e amazonica, apreendidos de tráfico de animais selvagens: 18 apresentavam ingluvite e 22 outras alterações, mas todos mostrando sinais de debilitação geral. Colheram-se as amostras clínicas através da introdução de sonda uretral no esôfago dos animais e estas foram semeadas em ágar Sabouraud dextrose acrescido de cloranfenicol. A identificação das espécies de Candida foi baseada em características macro e micromorfológicas e comportamento bioquímico no kit API 20C; pesquisou-se ainda a produção das enzimas proteinase e fosfolipase. Isolou-se Candida spp. de 57,5 por cento dos papagaios, sendo 72,2 por cento de aves com ingluvite e 45,5 por cento de aves com outras afecções. Isolaram-se 25 cepas de Candida, 60 por cento e 40 por cento respectivamente de animais com e sem ingluvite. As espécies isoladas foram: C. humicola (28 por cento), C. parapsilosis (24 por cento), C. guilliermondii (20 por cento), C. famata (20 por cento) e C. albicans (8 por cento). Os resultados obtidos demonstram que C. albicans não é a espécie mais freqüentemente isolada e é a primeira vez que se relata C. guilliermondii, C. famata e C. humicola causando infecção em papagaios. A maioria dos isolados apresentou filamentação (76 por cento) e produziu a enzima fosfolipase (68 por cento) e todas as cepas produziram proteinase. O encontro de Candida spp., produzindo seus fatores de virulência, reforça o papel patogênico dessas leveduras nos casos estudados.(AU)

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The hoatzin is the only known avian species with foregut fermentation. It is a primarily folivorous feeder and has a distended crop and lower/distal esophagus, which has evolved for the microbial fermentation of ingested feed. Crop samples collected from 10 individual animals from the Apure River area, Apure State, Venezuela were examined for the presence and density of methanogens using 16S rRNA gene clone libraries and real-time PCR prepared from pooled and individual PCR products. A total of 197 clones were examined, revealing 24 different methanogen 16S rRNA sequences, or phylotypes. Of the 24 unique phylotypes, 16 (171 of 197 clones) formed five unique clades within the genus Methanobrevibacter with the largest group of clones (118 clones) 98.7% similar to Methanobrevibacter ruminantium. The remaining eight phylotypes (26 clones) formed four unique clades that had only 94.0-96.7% identity to Methanosphaera stadtmanae. Based upon 98% sequence identity, we identified 17 of the 24 methanogen phylotypes from the hoatzin as possible new species and strains, with three phylotypes representing possible new genera (<94.5% sequence identity). Although none of the hoatzin methanogen phylotypes had 100% sequence identity to any other archaeal sequences in the GenBank database, the hoatzin crop methanogen sequences formed sister groups with known rumen methanogens. Mean population densities (numbers per gram wet weight) of methanogenic archaea, rumen bacteria and ciliate protozoa, estimated using real-time PCR, were 5.80 x 10(9), 7.93 x 10(12) and 3.31 x 10(5), respectively. The crop microbial data presented here provide an excellent example of convergent evolution of foregut fermentation in the hoatzin, similar to that of ruminants.

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To determine the inhibitory capacity of lactic acid bacteria due to the action of antagonistic substances, we tested 474 isolates of Lactobacillus from the crop and cecum of chickens against gram-positive and gram-negative indicator microorganisms by the spot-on-the-lawn and well-diffusion antagonism methods. Of the 474 isolates, 265 demonstrated antimicrobial activity against the indicator microorganisms. Isolates identified as L. reuteri, L. salivarius, or Lactobacillus spp. inhibited Enterococcus faecalis, E. faecium, Listeria monocytogenes, and Salmonella spp. but not L. casei, L. delbrueckii, L. fermentum, or L. helveticus by the well-diffusion simultaneous antagonism method under anaerobic incubation conditions. The antagonistic substances produced by some of the Lactobacillus isolates were inactivated after treatment by proteolytic enzymes, which suggested that the substances could be antimicrobial peptides or bacteriocins.

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