Investigation of Salmonella spp. in free-range chickens (Gallus gallus domesticus) in Southern Rio Grande do Sul / Pesquisa de Salmonella spp. em galinhas coloniais (Gallus gallus domesticus) no sul do Rio Grande do Sul

Salmonelose é uma doença causada por bactérias do gênero Salmonella, com importância para saúde pública e animal. Dentre os sorotipos hospedeiro-específicos, destaca-se o Gallinarum, que possui os biovares Gallinarum e Pullorum adaptados às aves e amplamente difundidos pelo mundo. Os dados sobre a ocorrência de Salmonella spp. em criações avícolas alternativas no Brasil são escassos. O objetivo deste estudo foi pesquisar a ocorrência de Salmonella spp. em galinhas coloniais encaminhadas para necropsia ao LRD/FV/UFPel. Foram realizadas análises histopatológicas, microbiológicas e moleculares das colônias bacterianas isoladas de 12 amostras de órgãos de galinhas domésticas dos municípios de Pelotas e Piratini, no Rio Grande do Sul. Na análise microbiológica, foram isoladas bactérias do gênero Salmonella sorotipo Gallinarum das 12 amostras, sendo 10/12 bioquimicamente compatíveis com biovar Gallinarum e 2/12 com biovar Pullorum. Na análise molecular PCR 11/12, 91,7% foram identificadas genotipicamente como Salmonella spp. O presente estudo demonstrou uma elevada frequência de isolamento de Salmonella Gallinarum biovar Gallinarum em aves sintomáticas criadas em regime extensivo. Além disso, os dados epidemiológicos das aves analisadas demonstram que a infecção por Salmonella Gallinarum nesses casos está associada ao contato com aves silvestres e falhas de manejo sanitário.(AU)

Escherichia coli and Salmonella ser. Saintpaul natural co-infection in a free-living ruddy ground dove (Columbina talpacoti): a case report / Coinfecção por Escherichia coli e Salmonella ser. Saintpaul em Columbina talpacoti: relato de caso

This study reports a co-infection of Escherichia coli and Salmonella in a free-living ruddy ground dove (Columbina talpacoti) received at the Laboratory of Ornithological Studies of the State University of Ceará, Brazil. The bird presented diarrhea, leg paralysis and anorexia, and died shortly after. Necropsy was then performed and samples from lung, kidney, liver and intestine were collected for microbiological and histopathological analyses. Escherichia coli was isolated from cloacal swab, lung and kidney samples. Salmonella ser. Saintpaul was identified in liver and spleen samples. Escherichia coli isolates were tested for the presence of eight diagnostic genes for diarrheagenic pathotypes (STEC, ETEC, EPEC, EIEC, EAEC) with conventional polymerase chain reaction (PCR). EAEC was detected in the lung and kidney, and STEC in the intestine. In conclusion, Columbina talpacoti is susceptible to enteroaggregative Escherichia coli and Salmonella ser. Saintpaul infection, which may have public health implications.(AU)

Este estudo relata um caso de coinfecção por Escherichia coli e Salmonella ser. Saintpaul em uma rolinha-roxa (Columbina talpacoti) recebida pelo Laboratório de Estudos Ornitológicos da Universidade Estadual do Ceará, Brasil. A ave apresentava diarreia, paralisia nas pernas e anorexia, indo a óbito rapidamente. A necropsia foi realizada e amostras de pulmão, rim, fígado e intestino foram coletados para isolamento microbiológico e análise histopatológica. Escherichia coli foi identificada em amostras de suabe cloacal, pulmão e rim. Salmonella ser. Saintpaul foi identificada no fígado e baço. Isolados de E. coli foram testados para a presença de oito genes de diagnóstico para patotipos diarreiogênicos (STEC, ETEC, EPEC, EIEC, EAEC) através de reação em cadeia de polimerase (PCR) convencional. EAEC foi detectada no pulmão e rim, e STEC foi identificada no intestino. Em conclusão, Columbina talpacoti é suscetível a infecção por Escherichia coli enteroagregativa e Salmonella ser. Saintpaul, o que pode implicar em risco para a saúde pública.(AU)

Diagnóstico de un brote de Salmonella Typhimurium en Chinchillas (Chinchilla lanigera) mediante la electroforesis en gel de campo pulsado / Diagnosis of an autbreak of Salmonella Typhimurium in chinchillas (Chinchilla lanigera) by pulsed-fiel gel electrophoresis

Empleando estudios anatomopatológicos y microbiológicos se examinó a un grupo de chinchillas (Chinchilla lanigera) adultas que murieron súbitamente en 2012 en una granja de la ciudad de La Plata (Buenos Aires, Argentina). Se aisló Salmonella enterica serovar Typhimurium (S. Typhimurium) del hígado, el bazo, el corazón, los pulmones, los riñones y los intestinos de los cinco animales evaluados. Los cinco aislamientos estudiados (uno por animal) fueron sensibles a ampicilina, cefalotina, cefotaxima, ácido nalidíxico, gentamicina, estreptomicina, cloranfenicol, fosfomicina, nitrofurantoína y trimetoprima-sulfametoxazol, y resistentes a tetraciclina. El análisis de dichos aislamientos por electroforesis en gel de campo pulsado [pulsed-field gel electrophoresis (PFGE)] con XbaI mostró un perfil electroforético idéntico con 15 bandas, idéntico a su vez al patrón ARJPXX01.0220 del banco nacional argentino de datos de PulseNet, que cuenta con patrones de PFGE de Salmonella. El presente trabajo describe por primera vez el diagnóstico postmortem de un brote de salmonelosis en chinchillas usando un método molecular, como la electroforesis en gel en campo pulsado

Adult chinchillas (Chinchilla lanigera) that had suddenly died in a commercial farm located in La Plata City, Buenos Aires Province, Argentina, in July 2012 were macroscopically, histopathologically, and microbiologically examined. Salmonella enterica serovar Typhimurium (S. Typhimurium) was isolated from the liver, spleen, heart, lungs, kidneys and intestines from each of the five animals evaluated. The five strains were susceptible to ampicillin, cephalotin, cefotaxime, nalidixic acid, gentamicin, streptomycin, chloramphenicol, fosfomycin, nitrofurantoin and trimethoprimsulfamethoxazole, and resistant to tetracycline. Each of the five S. Typhimurium isolates was analyzed by XbaI- pulsed-field gel electrophoresis (PFGE), showing an identical electrophoretic profile with 15 defined bands, which was found to be identical to pattern ARJPXX01.0220 of the PulseNet Argentine National database of Salmonella PFGE patterns. This is the first work describing the postmortem diagnosis of an outbreak of salmonellosis in chinchillas by using molecular methods such as PFGE

Detection of virulence-associated genes in Salmonella Enteritidis isolates from chicken in South of Brazil / Detecção de genes associados à virulência em cepas de Salmonella Enteritidis isoladas de frangos na região sul do Brasil

Salmonella spp. are considered the main agents of foodborne disease and Salmonella Enteritidis is one of the most frequently isolated serovars worldwide. The virulence of Salmonella spp. and their interaction with the host are complex processes involving virulence factors to overcome host defenses. The purpose of this study was to detect virulence genes in S. Enteritidis isolates from poultry in the South of Brazil. PCR-based assays were developed in order to detect nine genes (lpfA, agfA, sefA, invA, hilA, avrA, sopE, sivH and spvC) associated with the virulence in eighty-four isolates of S. Enteritidis isolated from poultry. The invA, hilA, sivH, sefA and avrA genes were present in 100% of the isolates; lpfA and sopE were present in 99%; agfA was present in 96%; and the spvC gene was present in 92%. It was possible to characterize the isolates with four different genetic profiles (P1, P2, P3 and P4), as it follows: P1, positive for all genes; P2, negative only for spvC; P3, negative for agfA; and P4, negative for lpfA, spvC and sopE. The most prevalent profile was P1, which was present in 88% of the isolates. Although all isolates belong to the same serovar, it was possible to observe variations in the presence of these virulence-associated genes between different isolates. The characterization of the mechanisms of virulence circulating in the population of Salmonella Enteritidis is important for a better understanding of its biology and pathogenicity. The frequency of these genes and the establishment of genetic profiles can be used to determine patterns of virulence. These patterns, associated with in vivo studies, may help develop tools to predict the ability of virulence of different strains.

Salmonella spp. estão entre os principais agentes causadores de doenças transmitidas por alimentos, e o sorovar Salmonella Enteritidis é o mais frequentemente isolado no mundo. A virulência de Salmonella spp. e a sua interação com o hospedeiro são processos complexos que envolvem fatores de virulência para sobreviver às defesas do hospedeiro. O objetivo deste estudo foi detectar genes de virulência em cepas de S. Enteritidis isoladas a partir de fontes avícolas no sul do Brasil. Ensaios de PCR foram desenvolvidos para a detecção de nove genes (lpfA, agfA, sefA, invA, hilA, avrA, sopE, sivH e spvC) associados à virulência em oitenta e quatro amostras de S. Enteritidis. Os genes invA, hilA, sivH, sefA e avrA estavam presentes em 100% dos isolados; lpfA e sopE estavam presentes em 99%; agfA em 96%; e o gene spvC estava presente em 92%. Foi possível caracterizar os isolados em quatro perfis genéticos distintos (P1, P2, P3 e P4), sendo P1 positivo para todos os genes; P2 negativo apenas para spvC; P3 negativo para agfA e P4 negativo para lpfA, spvC e sopE. O perfil de maior frequência foi P1, presente em 88% dos isolados. Apesar de todos os isolados pertencerem ao mesmo sorovar, foi possível observar variações na presença de genes associados à virulência entre os mesmos. A caracterização dos mecanismos de virulência circulantes na população de Salmonella Enteritidis é importante para um maior entendimento da sua biologia e patogenicidade. A frequência destes genes e o estabelecimento de perfis genéticos podem ser utilizados para determinar os padrões de virulência dos isolados. Estes padrões, associados a estudos in vivo, podem auxiliar na elaboração de ferramentas que permitam predizer a capacidade de virulência das diferentes cepas.

Salmonella typhimurium em linfonodos mesentéricos de ovinos ao abate / Presence of Salmonella typhimurium in ovine mesenteric lymph nodes at slaughter

O rebanho de ovinos no Brasil está estimado em mais de 16 milhões de cabeças. Embora o consumo da carne desta espécie ainda seja pequeno, comparado ao de outros países, o consumo de carne, inclusive ovina, tem sido associado às doenças transmitidas por alimentos, em especial a salmonelose. No presente estudo, investigou-se a ocorrência de salmonelas em linfonodos mesentéricos e conteúdo intestinal de 175 ovinos ao abate. "Pools" constituído por cinco amostras de contéudo fecal ou 5 amostras de linfonodos de 25 g foram pre-enriquecidos em 250 mL de água peptonada tamponada e incubados a 37° C por 18-24 horas. Uma alíquota de 0,1 mL do préenriquecimento foi transferida para 9,9 mL de caldo de enriquecimento Rappaport-Vassiliadis e 1,0 mL do pré-enriquecimento foi transferido para 10 mL de caldo tetrationato Muller-Kaufmann, incubados a 42° C for 24h. 10 µL do caldo de enriquecimento foi semeado superfície de placas de ágar BPLS e ágar XLT4 incubadas a 37º C for 24-48h. Colônias suspeitas de salmonela foram testadas por provas bioquímicas e serologicas. Os testes bioquímicos utilizados para identificação de Salmonella foram TSI (triple sugar iron àgar), LIA (lysine iron àgar) e ágar ureia. Sorotipagem foi realizada no Laboratório de Enterobactérias do Instituto Osvaldo Cruz. Isolou-se Salmonella Tiphymurium de um pool de linfonodos mesentéricos, provenientes de cinco animais. O fato de se observar a ocorrência de salmonela em ovino portador sadio alerta para necessidade de monitorar este micro-organismo também nesta espécie, especialmente quando destinada ao abate, com vistas à produção de alimentos seguros.

The ovine flock in Brazil is estimated at over 16 million head. Despite that meat consumption of this species is still small when compared to other countries, general meat consumption, including mutton, has been associated to food borne diseases, especially salmonellosis. In the present study, the occurrence of salmonella in mesenteric lymph nodes and intestinal content of 175 ovines during slaughter was investigated. A pool of 5 feces samples or 5 lymph node samples of 25 grams was pre-enriched in 250 mL of buffered peptone water at 37° C for 18-24h. Following this, 0.1 mL of pre-enriched broth was transferred to 9.9 mL of Rappaport-Vassiliadis enrichment broth and 1.0 mL of pre-enriched broth was transferred to 10 mL of Muller-Kaufmann tetrationate broth, incubated at 42° C for 24h. Then, a 10 µL of the enrichment broth was spread on the surface of a BPLS and an XLT4 plate, both incubated at 37º C for 24-48h. Suspected Salmonella colonies were picked from the agar and tested with biochemical and serological methods. Biochemical testing was carried out for the identification of Salmonella, using the TSI (triple sugar iron agar), LIA (lysine iron agar) and urea agar tests. Serotyping was done at the Laboratory of Enterobactérias of the Instituto Osvaldo Cruz. Salmonella Tiphymurium was isolated from a pool of mesenteric lymph nodes from 5 animals. That Salmonella was observed in healthy carrier ovines points out the necessity of monitoring this microorganism in this species as well, especially when animals are destined to slaughter, so to assure safe food production.

Métodos diagnósticos para os patógenos alimentares: Campylobacter sp., Salmonella sp. e Listeria monocytogenes / Diagnostic methods for the food pathogens Campylobacter sp., Salmonella sp. and Listeria monocytogenes

Os métodos moleculares de detecção rápida e eficaz de lotes de aves infectados por bactérias como Salmonella sp. Campylobacter sp. e Listeria monocytogenes são importantes para reduzir a frequência da transmissão destes patógenos entre os lotes de aves e aos consumidores de produtos de origem animal. Recentemente, as técnicas de biologia molecular, em especial a reação em cadeia polimerase, que permite a amplificação específica de segmentos de DNA, têm possibilitado novos rumos na identificação de bactérias supracitadas, reduzindo o tempo de cultivo e ampliando a confiabilidade das provas diagnósticas. A utilização da biologia molecular por laboratórios de diagnóstico humano e animal, assim como em programas de controle de qualidade de alimentos e produtos de origem animal, já é realidade e tende a se expandir rapidamente. O objetivo deste artigo é fazer uma breve revisão dos testes diagnósticos convencionais e moleculares para identificar Campylobacter sp., Salmonella sp. e Listeria monocytogenes. Concluindo, o diagnóstico molecular é um campo em avanço científico e tecnológico, no qual novas técnicas moleculares estão em desenvolvimento para o diagnóstico de bactérias em alimentos.

The molecular methods for quick and efficient detection of chicken lots infected by bacteria such as Salmonella sp. Campylobacter sp. and Listeria monoytogenes is basic for the effort to reduce the frequency of the transmission between chicken lots and to the consumers of poultry products. Recently, the development of techniques involving molecular biology, especially polymerase chain reaction, which allows the specific enlargement of segments of DNA, has been making new procedures possible for the identification of the abovementioned bacteria, reducing the time necessary for the tests and enhancing the reliability of the resulting diagnoses. The use of molecular biology in laboratories for human and animal diagnosis, as well as in quality control programs for foods and products of animal origin is already a reality and has tended to expand quickly. The objective of this article is to present a brief review of the conventional diagnostic and molecular tests for the identification of Campylobacter sp., Salmonella sp. and Listeria monocytogenes. In conclusion, molecular diagnosis is a field undergoing scientific and technological advancement, in which new molecular techniques are under development for the diagnosis of bacteria in foods.

Avaliação da reação em cadeia da polimerase e do isolamento bacteriológico convencional na detecção de Salmonella Dublin em amostras de fezes de bezerros infectados experimentalmente / Evaluation of polymerase chain reaction and standard microbiological techniques for detection of Salmonella Dublin in fecal samples of infected calves

The efficiency of the Polymerase Chain Reaction (PCR) combined with selective enrichment broth was compared with the standard microbiological techniques for detection of Salmonella Dublin in fecal samples of 10 to 15-days-old Holstein calves, experimentally infected with 10(8) CFU of Salmonella Dublin. Seventy-six fecal samples were analyzed using PCR associated with selenite cystine (SC) and Muller-Kauffmann tetrathionate (TMK) broths and standard microbiological techniques. Regardless of the selective enrichment broth used, the standard microbiological techniques were significantly better than PCR in detection of positive samples of Salmonella Dublin. However, the simultaneous use of both techniques provided detection of a larger number of positive samples. The SC broth was the best option as selective enrichment in both techniques.

Avaliação clínica da infecção experimental de bezerros com Salmonella Dublin / Clinical evaluation of experimental Salmonella Dublin infection in calves

The clinical conditions of healthy calves infected with experimental 10(8)CFU of Salmonella Dublin were evaluated and the viability of the experimental model in disease induction in calves was verified. Twelve 10 to 15-day-old male Holstein calves were examined. They were allocated into two groups, control and experimentally infected with 10(8)CFU of Salmonella Dublin. Animals were submitted to clinical examination after inoculation and at every 12 hours, during seven days after the experimental infection. Samples of rectal swabs were collected for Salmonella Dublin isolation. All animals had severe diarrhea, with mucus and bleeding, 12 to 84 hours after the experimental infection with Salmonella Dublin, accompanied by fever, dehydration and respiratory signs. The isolation of Salmonella Dublin from rectal swabs occurred 12 hours after the infection. Two out of the six animals inoculated with Salmonella Dublin died with symptoms of enteritis, fibrinous pneumonia, centrilobular hepatic steatosis, hepatocyte necrosis, spleen congestion, interstitial nephritis, and tubular degeneration. Thus, the oral administration of 10(8)CFU of Salmonella Dublin induced clinical signs of salmonellosis in 10 to 15-day-old calves.

Rapid detection of Salmonella sp. in pork samples using fluorescent in situ hybridization: a comparison with VIDAS®-SLM system and ISO 6579 cultural method / Detecção rápida de Salmonella sp. em amostras de suinos por hibridação in situ fluorescente: comparação com o sistema VIDAS®-SLM e com o método de cultura ISO 6579

This study reports the use of the fluorescent in situ hybridization (FISH) with Sal3 probe for Salmonella detection in swine carcasses inner surface (swab); and in the correspondent samples of ileum, ileocolic, and mandibular lymph nodes; and tonsils, after dilution (1:10) in buffered peptone water and a pre-enrichment step (37(0)C, 18h). In order to evaluate the efficiency of FISH, 235 naturally contaminated samples were simultaneously tested by the cultural method (ISO 6579) and by the Vitek Immuno Diagnostic Assay System (VIDAS®) - Salmonella (SLM) system. The cultural method identified 39 positive samples. From these, VIDAS®- SLM only detected 23. FISH identified 115 positive samples. This difference was highly significant (P<0.001). From positive samples, 32 were also confirmed by the cultural method. The results indicate FISH as a promising tool for rapid Salmonella detection in samples of pork and swine carcasses

Descreve-se a utilização da técnica de hibridação in situ fluorescente (FISH), utilizando a sonda Sal3, para detecção de Salmonella na superfície interna de carcaças de suínos (zaragatoa), em amostras correspondentes de íleo, linfonodos ileocólicos, linfonodos mandibulares e amígdalas, após terem sido diluídas (1:10) e submetidas a uma fase de pré-enriquecimento em água peptonada tamponada (a 37ºC, 18h). Para avaliar a eficácia do método FISH, analisaram-se 235 amostras naturalmente contaminadas, usando o método de cultura ISO 6579 e o sistema Vitek Immuno Diagnostic Assay System (VIDAS®)- Salmonella (SLM), simultaneamente. O método de cultura identificou 39 amostras positivas, das quais o método VIDAS®-SLM detectou apenas 23. O método FISH identificou 115 amostras positivas. A diferença entre os métodos foi altamente significativa (P<0.001). Das amostras positivas, 32 foram confirmadas pelo método de cultura. Os resultados indicam que a FISH constitui uma promissora técnica de detecção rápida de Salmonella em amostras de suínos abatidos para consumo

Salmonella sp. em avicultura industrial: diagnóstico imunológico e molecular / Salmonella sp. in poultry industry: immunological and molecular diagnostics

Os sorotipos de salmonelas paratifóides, tais como, Salmonella enteritidis e Salmonella typhimurium acometem diversas espécies de animais e possuem significado em saúde pública, como agentes etiológicos da salmonelose humana.A indústria avícola atende a uma série de exigências para o controle higiênico-sanitário dos plantéis e produtos avícolas. A legislação inclui procedimentos de campo, de abate e de processamento, rotulagem, diagnóstico laboratorial e certificação de produtos, a fim de evitar a transmissão de Salmonella sp. para outros animais e seres humanos. As tecnologias de diagnóstico de Salmonella sp. confiáveis e rápidas são essenciais para que a indústria avícola atenda às exigências legais e comerciais e diminua o tempo de estocagem dos produtos em câmaras e contêineres frigoríficos. Esta revisão objetiva abordar os principais assuntos relacionados ao diagnóstico imunológico e molecular de Salmonella sp. e destacar a importância do diagnóstico rápido deste patógeno para a indústria avícola nacional.

Simultaneous detection of Lawsonia intracellularis, Brachyspira hyodysenteriae and Salmonella spp. in swine intestinal specimens by multiplex polymerase chain reaction

A multiplex PCR assay was developed for the simultaneous detection of the etiologic agents associated with porcine proliferative enteropathies (PPE), swine dysentery (SD)and porcine salmonellosis (PS)in a single reaction using DNA from swine intestinal samples. Single and multiplex PCR amplification of DNA from Lawsonia intracellularis, Salmonella typhimurium and Brachyspira hyodysenteriae with each primer set produced fragments of the predicted size without any nonspecific amplification, 210-bp, 298-bp and 403-bp bands, respectively. The single PCR assay could detect as little as 100 pg of purified DNA of S. typhimurium and L. intracellularis, and 50 pg of B.hyodysenteriae, respectively. However, multiplex PCR turned out to be 10 times lower sensitivity with S. typhimurium compared with single PCR. With 23 swine intestinal specimens suspected of having PPE, SD and/or PS, the multiplex PCR assay showed identical results with conventional methods except one. In conclusion, this multiplex PCR is a feasible alternative to standard diagnostic methods for detection of L. intracellularis, B. hyodysenteriae and Salmonella spp. from swine intestinal specimens.

Comparison of different selective enrichment steps to isolate Salmonella sp. from feces of finishing swine

A two-phase study was conducted to compare the efficacy of several enrichment selective-broth steps associated to different plating media for recovery of Salmonella sp. from finishing swine feces. In a first phase, Rappaport-Vassiliadis broth (RV) incubated at 42liC, Tetrathionate Mseller-Kauffmann broth at 37liC (TMK37) and 42liC (TMK42), and Selenite Cystine broth (SC) at 37liC, in combination with three selective plating media Rambach agar (RA), Xylose-Lysine-Tergitol 4 agar (XLT4), and Brilliant-Green Phenol-Red Lactose Sucrose agar (VB) were compared for recovery of Salmonella from artificially contaminated swine feces. In a second phase, RV, TMK37, and TMK42, associated with XLT4 and VB , were tested with naturally contaminated swine feces. In this study RV, TMK42 and TMK37 were superior to SC for isolating Salmonella sp. from artificially contaminated feces. TMK42 and RV were more productive than TMK37 for recovery of Salmonella from naturally contaminated feces samples. Selectivity and indication capability of the plating media were remarkably affected by the selective enrichment step effectiveness. The TMK42/XLT4 association was the most sensitive and RV/XLT4 the most specific. The use of VB agar is also recommended to increase the likelihood of isolating atypical H2S-late producing/ non-producing Salmonella. In this study RV and TMK42 were the most efficient selective enrichment for recovery of Salmonella sp. from swine feces.

Differential diagnosis of Salmonella gallinarum and S. pullorum using PCR-RELP

Salmonellosis in poultry of Korea is a significant health problem, which causes substantial economic losses. The most common causative agents of chicken salmonellosis ar S. gallinarum and S. pullorum. Traditional methods used to detect Salmoenella spp. In chicken are tedious, time consuming and confer little guarantee of sensitivity and species specificity. Therefore, a rapid and sensitive method for the differentiation of Salmonella serogroup D was assessed. We first amplified the rfbS genes by PCR and characterized the amplified product by nucleotide sequence analysis. The homology of nucleotide sequence was 99.7% between S. gallinarum and S. pullorum rfbS genes. Further comparisons of the sequences of S. gallinarum, S. gallinarum fied strain, S. pullorum and S. typhi(GenBank Accession No.M29682) showed a homology of 98.3%. The predicted amino acid sequence homology was 97.1%, 97.1% and 97.5%, respectively. Based on this comparison of these nucleotide sequences, we found unique restriction enzyme sites within the rfbS genes of S. gallinarum and S. pullorum. Thus, the PCR products could be further digested with restriction enzymes TfiI and PleI for use in a restriction fragment length polymorphism (RELP) technique. This method can be applied in the differential diagnosis between S. gallinarum and S. pullorum.

Freqüência de parasitas e infecçäo por Salmonella em lobos guará, Chrysocyon brachyurus, mantidos em zoológicos no Estado de Säo Paulo / Frequency of parasites and Salmonella infection in captive maned-wolf, Chrysocyon brachyurus, kept in Zoos at the State of Säo Paulo, Brazil

Trinta e um lobos guará (Chrysocyon brachyurus, Illiger 1815), mantidos em 11 zoológicos no Estado de Säo Paulo, foram estudados para verificar a presença de parasitas e de infecçäo por Salmonella usando-se métodos de diagnóstico parasitológico e cultura seletiva de fezes. Os ecto e endoparasitas encontrados foram: Ctenocephalides felis (56,2 por cento), Rhipicephalus sanguineus (12,5 por cento), Ancylostoma caninum (45,l por cento), Strongyloides sp. (29,0 por cento), Uncinaria stenocephala (3,2 por cento), Capillaria sp. (3,2 por cento), Entamoeba sp. (22,9 por cento), Sarcocystis sp. (29,0 por cento), Cryptosporidium sp. (19,3 por cento), Eimeria sp. (19,3 por cento), Giardia sp. (9,6 por cento) e Isospora sp. (3,2 por cento). Seis animais apresentaram infecçäo por Salmonella e quatro diferentes sorotipos foram identificados. Somente um animal apresentava quadro de diarréia sugerindo que a capacidade de portar Salmonella spp. como flora gastrintestinal näo patogênica parece ser uma adaptaçäo fisiológica dessa espécie animal

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

The effects of treatment with anaerobic cecal microflora (ACM) and/or lactose and/or acetic acid on systemic and digestive tract of broiler chicks infection with "Salmonella" Typhimurium and "S." Enteritidis were studied. ACM was used without previous bacterial identification. Treatment with ACM contributed to the resistance of broiler chicks to infection with "Salmonella" spp. The infections were more persistent in the cecum, rectum and crops in decreasing order of intensity. The infections were also self-limiting since treated and control lots presented similar infection rates at the end of the experiments. Alone or in combination with lactose, ACM reduced the colonization of the digestive tract of broiler chicks by "S." Typhimurium and "S." Enteritidis. The effect of the combination of ACM with lactose or acetic acid was not potentiated in terms of reduction of fecal excretion of "Salmonella" spp. Treatment with ACM reduced the amount of "S." Typhimurium and "S." Enteritidis in feces. Alone or in combination with lactose, ACM reduced the cecal pH in treated birds. "S." Enteritidis was much more invasive than "S." Typhimurium and the use of ACM alone was more effective on the reduction of systemic infection. An explanation for the process of prevention of intestinal colonization with "Salmonella" spp. probably resides in the interrelationship of physiological, microbiological and immunological phenomena, as well as the variation in cecal pH.

Prevention of Salmonella infection by contact using intestinal flora of adult birds and/or a mixture of organic acids

This study was carried out to assess the ability of competitive exclusion and a mixture of organic acids to prevent "Salmonella" infection by contact between newly hatched chicks. A bird infected with "Salmonella" was placed in a box containing non-infected birds, previously treated with a broth culture of faeces of adult birds (CE) and/or a mixture of organic acids. The number of "Salmonella" organisms in the caeca of the contact birds was estimated at 4 and 8 days post-challenge. The birds were infected with "Salmonella" Typhimurium, "Salmonella" Enteritidis (both repeated 5 times), "Salmonella" Agona and "Salmonella" Infantis (3 repetitions). The same approach was used to test the mixture of organic acids alone. In this case the birds recieved feed containing 0.8(per cent) of a mixture of formic acid (70(per cent)) and propionic acid (30(per cent)). Also, a third trial was carried out with birds inoculated with the broth culture of faeces and fed with feed containing the mixture of organic acids. Appropriate controls were included. Whereas the birds from the control groups and the groups treated with the mixture of organic acids were heavily infected with "Salmonella", those pre-treated with CE or CE plus the mixture of organic acids had no viable cells per gram of caecal contents.

Detection of Salmonella contamination in food samples by dot-ELISA, DNA amplification and bacterial culture.

A dot-blot enzyme-linked immunosorbent assay (dot-ELISA) employing a genus Salmonella specific monoclonal antibody (MAb) was used for detection of the bacteria in food samples in comparison with the conventional culture method and the DNA amplification. Among the 200 chicken and pork samples (100 each) tested, 9% and 33%, 7% and 20% and 7 and 23% were positive for salmonellae by the dot-ELISA, the culture method and the DNA amplification, respectively. Statistical analyses revealed that the sensitivity, specificity, efficacy, and positive and negative predictive values of the detection of Salmonella in the food samples by dot-ELISA compared with the culture method were 93.33%, 91.76%, 92%, 66.66% and 98.73%, respectively. Comparison of the DNA amplification and the culture method revealed the sensitivity, specificity, efficacy, and positive and negative predictive values of 100%, 91.58%, 92%, 65.21% and 100%, respectively. The dot-ELISA and the DNA amplification results were in a better agreement when the two assays were compared. The sensitivity, specificity, efficacy, positive and negative predictive values of the dot-ELISA compared to the DNA amplification were 91.3%, 100%, 98%, 100% and 97.5%, respectively. From this study, the dot-ELISA is rapid, simple, sensitive, specific at low cost with limited amount of infectious waste to be disposed and offers another advantage in that it detects only the smooth LPS of Salmonella which implies the possible presence of the virulent organisms.