A doença do edema afeta os leitões desmamados e é causada por cepas de Escherichia coli adaptadas ao hospedeiro e produtoras da toxina Stx2e. A doença do edema é caracterizada por edema de pálpebras, ataxia, pedalagem, dificuldade locomotora, e morte súbita. No presente estudo foram avaliadas 158 cepas de E. coli positivas para presença do gene codificador da toxina Stx2e isoladas de 62 animais provenientes de 13 granjas localizadas nos Estados do Rio Grande do Sul, Santa Catarina, Mato Grosso, Paraná, São Paulo, Goiás e Minas Gerais. As cepas foram submetidas a determinação do perfil de resistência, caracterização dos genes de virulência, determinação do grupo filogenético, expressão da toxina em cultivos de célula Vero, caracterização genotípica através da eletroforese em campo pulsado (PFGE) e polimorfismo do comprimento de fragmentos amplificados com uma única enzima (SE-AFLP). Dentre as 158 cepas stx2e+ foram identificadas 83,5% (132/158) apresentando resistência a três classes de antimicrobianos ou mais. Altos níveis de resistência forma identificados contra tetraciclina, sulfonamidas e florfenicol. A frequência dos genes de virulência associados a doença do edema como a fímbria F18, por exemplo, foi baixa em relação a outros estudos. Todas as 158 cepas testadas apresentaram a expressão da toxina e efeito citotóxico em células Vero. A caracterização dos grupos filogenéticos permitiu a distribuição das cepas nos quatro grupos descritos a seguir: grupo A 27,2% (43/158), grupo B1 3,8% (6/158), grupo B2 39,2% (62/158) e grupo D 29,8% (47/158). As cepas foram caracterizadas através da PFGE e do SE-AFLP e ambas as técnicas apresentaram altos índices discriminatórios (0,98 e 0,99 respectivamente). A associação mais frequente nas duas técnicas genotípicas foi observada em relação ao animal e a granja de origem. Apesar de pertencerem a um patotipo definido (STEC) e de serem altamente especializadas em relação ao hospedeiro, o suíno, foi observada uma grande variabilidade genética e de perfis de resistência entre as cepas de E. coli estudadas. / Edema disease affects weaning piglets and is caused by a host adapted Escherichia coli and producer of Stx2e toxin. The edema disease is characterized by swollen eyelids, ataxia, recumbence, convulsions, paralysis, or sudden death. In the present study were evaluated 158 E. coli strains Stx2e toxin gene positive isolated from 62 animals, from 13 swine herds located at Rio Grande do Sul, Santa Catarina, Mato Grosso, Paraná, São Paulo, Goiás and Minas Gerais States. Strains were submitted to resistance profile determination, virulence genes detection, phylogenetic group characterization, toxin expression in Vero cell culture, genotypic characterization through pulsed field gel electrophoresis (PFGE) and single enzyme amplified fragments length polymorphism (SEAFLP). Among the 158 strains stx2e+ were identified 83.5% (132/158) resistant to more than three or more classes of antimicrobials, High levels of resistance were found against tetracycline, sulfonamide and florfenicol. The frequency of virulence genes associated to edema disease as F18 fimbriae, for example, was low in relation to other studies. All 158 tested strains presented Stx2e toxin expression and cytotoxic effect in Vero cell culture. The characterization of phylogenetic groups permitted the distribution of strains in four groups described as follow: group A 27.2% (43/158), group B1 3.8% (6/158), group B2 39.2% (62/158) and group D 29.8% (47/158). The strains were characterized through PFGE and the SE-AFLP, and both techniques presented high discriminatory index (0.98 and 0.99 respectively). The association more frequently in both techniques was observed in relation to animal and herd origin. Despite to belong to one defined pathotype (STEC) and to be highly specialized in relation to host, the swine, it was observed a high variability of genetic and resistance profile among tested E. coli.

Pesquisas sobre Listeria monocytogenes em indústrias de produtos lácteos no Brasil são escassas. Três laticínios (A, B e C) produtores de queijos do Estado de São Paulo foram monitorados para a presença de L. monocytogenes no período de outubro/2008 a setembro/2009. Foram realizadas 12 coletas, correspondentes a 12 lotes de queijo produzidos, sendo quatro de cada laticínio. Em cada laticínio, as visitas foram realizadas com intervalos de aproximadamente 2 meses entre cada uma. Foram analisadas 393 amostras, sendo 201 de superfícies com e sem contato com alimentos e 192 de alimentos (leite cru e pasteurizado e queijo) água e salmoura, para pesquisa de L. monocytogenes. As análises foram realizadas de acordo com o método do Food and Drug Administration (FDA). Os resultados confirmam a presença de Listeria spp nas instalações dos três laticínios. L. monocytogenes, L. innocua, L. seeligeri e L. welshimeri foram as espécies isoladas neste estudo. Especificamente a espécie L. monocytogenes não foi encontrada no laticínio A, entretanto, o microrganismo foi isolado de 12,5% das amostras do laticínio B e de 9,1% do laticínio C. L. monocytogenes não foi isolada do leite cru dos silos, do leite pasteurizado, da água e dos queijos Minas frescal, nos 3 laticínios. Porém, no laticínio C, L. monocytogenes foi isolada de amostras de queijo Prato que foram incluídas apenas na 4ª coleta deste laticínio, além de ter sido isolada de amostras de salmoura. As maiores prevalências de contaminação por L. monocytogenes ocorreram em superfícies sem contato com alimentos, sendo positivas 51,6% das amostras do laticínio B e 21,7% do laticínio C. Em ambos os laticínios a bactéria também foi isolada de superfícies com contato com alimentos. Os resultados fornecem informações detalhadas dos pontos prioritários para o desenvolvimento de estratégias de controle de L. monocytogenes em laticínios e mostram a importância de programas de monitoramento ambiental do patógeno, mesmo em pequenas indústrias. Os 85 isolados identificados como L. monocytogenes revelaram-se de quatro sorotipos: 1/2a, 1/2b, 1/2c e 4b, com predomínio do 4b, em ambos os laticínios, o que é preocupante para a saúde pública. Com base nos resultados de PFGE (perfis combinados ApaI e AscI), 40 perfis (pulsotipos) foram obtidos. Pulsotipos foram isolados repetidamente entre coletas nos laticínios B e C, sugerindo persistência de linhagens nos laticínios. Apesar dos laticínios serem distantes e independentes, um pulsotipo foi compartilhado entre ambos. O laticínio A apresentou contaminação por mais de um pulsotipo de L. seeligeri e houve isolamento repetido de um pulsotipo dessa espécie, entre as coletas, sugerindo adaptação da bactéria e necessidade de controle do gênero Listeria nessa indústria. A ocorrência de um mesmo pulsotipo de L. monocytogenes com sorotipos diferentes (1/2b e 4b) mostra que a sorotipagem deve acompanhar análises mais refinadas como as de natureza genotípica. / Listeria monocytogenes surveys in cheese manufacturing plants in Brazil are rare. Three cheese manufacturing plants (A, B and C) in São Paulo state were monitored for the presence of Listeria monocytogenes during the period of October/2008 - September/2009. Twelve samples surveys were taken corresponding to 12 cheese lots produced, four in each plant. In each cheese plant, the samples were taken at intervals of approximately 2 months. There were 393 samples analyzed, 201 from surfaces with and without contact with food and 192 of food (raw and pasteurized milk and cheese), water and brine, with the objective of searching for L. monocytogenes. The analyses were performed in accordance with Food and Drug Administration (FDA) method. The results confirmed the presence of Listeria spp in the facilities of three plants. L. monocytogenes, L. innocua, L. seeligeri and L. weshimeri were the species isolated in this study. Specifically the L. monocytogenes specie was not isolated from plant A. However, the microorganism was isolated in 12.5% of the samples from plant B and 9.1% from plant C. Listeria monocytogenes was not isolated from raw milk in storage tanks, pasteurized milk, water or Minas frescal cheese samples from the three plants. Nevertheless, in plant C, L. monocytogenes was isolated in Prato cheese that was included only in the 4th sampling survey and also from the brine samples. The major prevalence of contamination by L. monocytogenes occurred on surfaces without contact with food, with 51.6% of the samples positive from plant B and 21.7% from plant C. In both plants, the microorganism was also isolated from food contact surfaces. The results provide detailed information about the critical points for the development of L. monocytogenes control strategies in cheese processing plants and, moreover, show the relevance of sampling programs of the pathogen, even in small cheese processing plants. The 85 isolates identified as L. monocytogenes were classified in four serotypes: 1/2a, 1/2b, 1/2c and 4b, with 4b dominating in both cheese plants, which is of concern to human health. On the basis of PFGE results (combined profiles ApaI and AscI), 40 profiles (pulsotypes) were found. Pulsotypes were isolated repeatedly among sampling surveys in plants B and C, suggesting persistence of lineages in the plants. Despite these plants being distant and independent, one pulsotype was shared between them. Plant A presented contamination by more than one pulsotype of L. seeligeri and there was a repetitive isolation of one pulsotype of this specie among samplings, suggesting adaptation of the bacterium and the need for control of the Listeria genus in this plant. The occurrence of one single pulsotype of L. monocytogenes with different serotypes (1/2b and 4b) show that serotyping should follow more refined analyses as the ones of genotypic nature.

Entre os microrganismos relevantes para a segurança alimentar, bactérias do gênero Salmonella tem se destacado como causadoras de toxinfecções, sendo motivo de preocupação constante para a cadeia de produção de aves e suínos. Os objetivos deste trabalho foram estudar a ocorrência de Salmonella spp. em suínos sadios ao abate e em animais apresentando sinais clínicos de salmonelose, tipificação dos isolados através da sorotipagem, polimorfismo do comprimento de fragmentos amplificados (AFLP) e eletroforese em gel de campo pulsado (PFGE). Foram examinados 50 animais com sintomatologia sugestiva de salmonelose de 12 granjas dos Estados de São Paulo, Paraná e Rio Grande do Sul e analisadas amostras de fezes, linfonodos e suabes de carcaças de 124 animais de quatro frigoríficos do Estado de São Paulo. Dos suínos com sintomatologia clínica, 38% dos animais foram positivos para o isolamento de Salmonella spp., sendo selecionadas 45 cepas, classificadas como S.Typhimurium (29/45), S. Choleraesuis (9/45), S. Infantis (3/45) e S. enterica subsp. enterica (4/45). Dos 124 suínos sadios, 16,12% foram positivos para o agente, sendo isoladas 39 cepas que foram classificadas como S. London (11/39), S. Anatum (11/39), S. Typhimurium (8/39), S. Agona (2/39), S. Enteritidis (2/39) e S. enterica subsp. enterica (5/39). Através do AFLP, a caracterização genética dos isolados revelou índice discriminatório igual a 0,85 e gerou 12 perfis distintos. O índice discriminatório do PFGE foi 0,96 apresentando 31 padrões distintos. Ambas as técnicas possibilitaram uma boa correlação entre isolados, seus sorotipos e locais de isolamento, sugerindo grande potencial para sua aplicação na genotipagem de Salmonella spp. / Among relevant microorganisms to food security, Salmonella has been highlighted as a major cause of food borne diseases, being a constant concern for poultry and pig production chain. The goal of this study was to investigate the Salmonella spp incidence in healthy pigs at slaughter, as well as in pigs showing salmonellosis clinical signs, characterize strains by serotyping, Amplified fragment length polymorphism (AFLP) and Pulsed field gel electrophoresis (PFGE). Fifty animals, presenting salmonelosis suggestive clinical symptoms, from 12 swine herds from Sao Paulo, Parana and Rio Grande do Sul states were examined and clinical materials such as stool samples, lymph nodes and carcass swab from 124 animals from four Slaughterhouses in Sao Paulo state were analyzed. Among the pigs with clinical symptoms, 38% of the animals were positive for the Salmonella spp. isolation. Forty five strains were selected and classified as S.Typhimurium (29/45), S. Choleraesuis (9/45), S. Infantis (3/45) and S. enterica subsp. enterica (4/45). From the 124 healthy pigs studied, 16.12% were positive for the agent. Thirty nine strains were isolated and classified, by serotyping, as S. London (11/39), S. Anatum (11/39), S. Typhimurium (8/39), S. Agona (2/39), S. Enteritidis (2/39) and S. enterica subsp. enterica (5/39). Through AFLP, genetic characterization of isolates showed discriminatory index of 0.85 and generated 12 distinct profiles. The PFGE discriminatory index was 0.96, showing 31 distinct patterns. Both techniques allowed a good correlation between the isolates, its serotypes and isolation locations, suggesting great potential of its application in genotyping of Salmonella spp.

Ovinos são considerados reservatórios de Escherichia coli produtora da toxina Shiga (STEC). Este estudo pesquisou amostras de 100 carneiros no Estado de São Paulo. PCR foi empregada para detecção de stx1, stx2, eae, ehx, e intiminas. RFLP-PCR foi realizado para identificação das variantes de stx1/stx2 e fliC. Cinco isolados eae+stx- de sorotipos O128:H2, O145:H2, O153:H7 e O178:H7 apresentaram intiminas b, g e e. Cinqüenta e seis STEC foram obtidos e os genótipos foram stx1cstx2d-O118 (56,1%), stx1c (33,3%), stx2d-O118 (7,6%), e stx1cstx2dOX3a (3%). Os sorotipos mais freqüentes foram ONT:H8, ONT:H-, O75:H-, O174:H8 e O5:H-. Gene fliC codificando para H2, H8, H14, H16, H19, H21 ou H28 foi identificado em 25/33 isolados. ehx foi detectado em três/cinco EPEC Atípicas e em 38 (57,6%) STEC. Sensibilidade aos antimicrobianos foi observada em 77,2% dos isolados STEC. A análise da similaridade genética por PFGE revelou 24 perfis entre 40 isolados STEC e EPEC atípicas. O estudo demonstrou que ovinos saudáveis podem se comportar como reservatórios de STEC e EPEC atípica. / Lambs are reported as carriers of Shiga toxin-producing Escherichia coli (STEC) worldwide. This study surveyed 100 sheep in São Paulo, Brazil. PCR was used for detection of stx1, stx2, eae, ehx, and intimins. RFLP-PCR investigated the stx1/stx2 variants and flagellar antigen (fliC). Five isolates were eae+/stx-, belonging to serotypes O128:H2, O145:H2, O153:H7 and O178:H7, harboring b, g and e intimins. Fifty-six STEC isolates were recovered, and stx genotypes were stx1cstx2d-O118 (56.1%), stx1c (33.3%), stx2d-O118 (7.6%), and stx1cstx2dOX3a (3%). A diversity of STEC serotypes was observed, but most frequent were ONT:H8, ONT:H-, O75:H-, O174:H8 and O5:H-. fliC gene coding for H2, H8, H14, H16, H19, H21 or H28 was identified in 25/33 isolates. ehx gene was detected in three Atypical EPEC and in 38 (57,6%) STEC. Fifty-one (77.2%) STEC were susceptible to antimicrobials tested. Pulsed field gel electrophoresis (PFGE) revealed 24/40 profiles. The study showed that healthy ovine can be carrier of STEC and atypical EPEC in Brazil.

Haemophilus parasuis é um dos agentes bacterianos que tem assumido grande importância na indústria suinícola nos últimos anos. Por muitos anos foi considerada uma doença esporádica de suínos jovens, no entanto, com o surgimento de doenças imunossupressoras ou com o aumento da criação de suínos com alto status sanitário, sua freqüência vem assumindo proporções cada vez maiores. A caracterização das cepas através de métodos fenotípicos como a sorotipagem não tem sido suficiente para os estudos epidemiológicos sobre esta bactéria, uma vez que muitos isolados não são sorotipificáveis. Os objetivos deste estudo foram caracterizar os isolados através da sorotipagem, do Polimorfismo do Comprimento de Fragmentos Amplificados (AFLP) e da Eletroforese em Gel de Campo Pulsado (PFGE), comparando os resultados obtidos. Dentre as 51 cepas de Haemophilus parasuis avaliadas uma foi classificada como sorotipo 2, doze como sorotipo 4, seis como sorotipo 5, quatro como 13 e sete como sorotipo 14, sendo que vinte e uma amostras não foram sorotipificáveis. Através do AFLP foi possível caracterizar todos os isolados, com um índice discriminatório de 0,98, porém esta técnica não demonstrou uma boa reprodutibilidade. Através da PFGE utilizando a enzima Sma I apenas 14 cepas foram genotipadas, porém com a enzima Not I, todas apresentaram padrões de bandas e o índice discriminatório obtido foi de 0,95. Os perfis obtidos através da PFGE com a enzima Not I apresentaram a melhor correlação com os dados de origem das cepas e seus sorotipos, indicando que a técnica possui um bom potencial para aplicação em estudos epidemiológicos. / In recent years, Haemophilus parasuis has had a growing impact on the swine industry. In the past, Haemophilus parasuis infections were considered to be sporadic in young pigs. However, the incidence is increasing due to immunosuppressive diseases and the increase in the production of pigs with high sanitary status. Characterization of strains using methods based on phenotype identification is not enough to perform epidemiological studies on thebacterium, since a large percentage of isolates are nontypeable). The aim of this study was to characterize isolates according to serotyping, Amplified Fragment Length Polymorphism (AFLP), and Pulsed Field Gel Electrophoresis (PFGE), and to compare characterization results. Out of the 51 strains of Haemophilus parasuis evaluated in this study: one was serotype 2; twelve serotype 4; six serotype 5; four serotype 13; and seven serotype 14. Serotyping could not be performed on the remaining 21 samples. All isolates were characterized using AFLP, with a discriminatory index of 0.98, but reproducibility of this technique is not good. Regarding PFGE, 14 strains were genotyped using enzyme Sma I; yet, with enzyme Not I, all strains presented band size and discriminatory index power of 0.95. The profiles obtained using PFGE with Not I presented better correlation with the origin and serotype of the strains, suggesting that this technique could be used in epidemiological studies with promising results.

Pasteurella multocida é o agente causador de muitas doenças de importância econômica em medicina veterinária e tem sido descrita como um agente de alto potencial zoónotico. No Brasil, ao contrário do que se observa na literatura internacional, informações sobre a frequência deste agente em animais de companhia como cães, gatos e coelhos são inexistentes. O presente estudo teve como proposta isolar cepas de Pasteurella multocida a partir de cães, gatos e coelhos, avaliar a freqüência deste agente nestas espécies animais, caracterizar os isolados de acordo com o tipo capsular e genes codificadores de fatores de virulência através da reação em cadeia pela polimerase, avaliar o perfil de resistência a antimicrobianos dos isolados obtidos e caracterizar as amostras através da PFGE. Foram examinados 640 animais, sendo 191 gatos, 309 cães e 140 coelhos. Dentre os animais examinados 8,1% foram positivos para o isolamento de P. multocida, sendo 10,4% dos gatos, 0,9% dos cães e 20,7% dos coelhos avaliados. Dentre as 93 cepas selecionadas 22,5% foram sensíveis a todas as drogas testadas, 77,4% das cepas foram resistentes a pelo menos uma droga utilizada e 5,3% foram apresentavam resistência a três ou mais drogas. Através da PFGE foram observados 39 pulsotipos com índice discriminatório igual a 0,97. Todos os genes codificadores de fatores de virulência foram detectados em pelo menos um isolado, sendo que nenhum destes esteve presente em 100% das cepas avaliadas. / Pasteurella multocida is the causative agent of many diseases of economic importance in veterinary medicine and has been described as an agent a high zoonotic potential. In Brazil, contrary to what is observed in the international literature, information about the frequency of this agent in animals such as dogs, cats and rabbits are nonexistent. This study proposes isolate strains of Pasteurella multocida from dogs, cats and rabbits, evaluate the frequency of this agent in these animal species, characterize the isolates according to the capsular type, and genes encoding virulence factors through the reaction polymerase chain, evaluate the antimicrobial resistance profiles of isolates and characterize the samples by PFGE. We examined 640 animals, 191 cats, 309 dogs and 140 rabbits. Among the animals examined were positive 8.1% for the isolation of P. multocida, and 10.4% of cats, dogs 0.9% and 20.7% of the rabbits studied. Among the 93 selected strains 22.5% were susceptible to all drugs tested, 77.4% of strains were resistant to at least one drug were used and 5.3% showed resistance to three or more drugs. Through PFGE were observed 39 pulsotipos discriminatory index equal to 0.97. All genes encoding virulence factors were detected in at least one isolated, none of which was present in 100% of the strains evaluated.

Flavobacterium columnare is a freshwater fish bacterium responsible for columnaris disease, the second leading cause of mortality in pond raised catfish in the southeastern United States. Pulsedield gel electrophoresis (PFGE) is a particularly powerful tool in epidemiology and is now regarded as the gold standard for molecular typing of microorganisms. We developed methods for conducting PFGE on F. columnare, and determined its efficacy for characterizing F. columnare strains isolated from different locations in the Southeastern United States. Virulence diversity was observed in two different immersion challenge experiments conducted with 16 different isolates in channel catfish fingerlings. A direct correlation was found between the PFGE clustered groups and virulence. In summary, our results suggest that two genetic divisions of F. columnare channel catfish isolates exist, one that contains strains that are “primary” pathogens of channel catfish (Group A), and another that are “secondary” or opportunistic pathogens of catfish (Group B).

Thèse numérisée par la Direction des bibliothèques de l'Université de Montréal.

The genus Salmonella, comprised of 2400 serotypes, is one of the leading causes of foodborne illnesses in the US and has been used for the deliberate contamination of food. A rapid system for detection, isolation, typing and antibiotic susceptibility profiling is essential for diagnosis and source tracking in natural outbreaks or a bioterrorism event. Pure culture is essential for molecular typing and antibiotic resistance testing. The virulence and the resistance mechanisms of Salmonella are rapidly evolving and many are still unexplained. The first aim of the study was to rapidly detect and isolate Salmonella from intentionally contaminated food. The second aim was to build a DNA fingerprinting database for accurate identification of the subtype. The third objective was to study the antibiotic susceptibility patterns and the underlying mechanisms of resistance. A correlation between the DNA subtypes and antibiograms was hypothesized. An association between the resistance determinants and pathogenicity genes was expected. A total of 114 isolates including environmental and clinical sources were tested. General and selective enrichments and immunomagnetic separation (IMS) were tested for rapid detection and isolation of Salmonella from eight food groups. Isolates were subtyped by pulsed field gel electrophoresis (PFGE) and automated RiboPrinter®. Resistance to 31 drugs was tested by the Sensititre® system and integrons were identified by PCR. The association between virulence and resistance was verified by Southern hybridization. Of the three genes tested, ompF was found to be the most reliable target for identifying Salmonella subspecies I, III and IV. Detection by real time PCR after enrichment in buffered peptone water and isolation by IMS provided the fastest results. Sixty two ribotypes and 74 pulsotypes were observed for the 100 isolates subtyped. Sixty isolates were resistant to one or more antimicrobials and 12 had class-1 integrons. In conclusion, pure culture was achieved in 25 hours by IMS. Ribotyping, a comparatively rapid technique was found to be ideal for initial identification. PFGE, which was more discriminatory, was appropriate for source tracking. Contrary to the original hypothesis, no correlation between subtyping and antibiograms was observed and no association of integrons with the virulence genes tested was demonstrated

The bacterium Listeria monocytogenes can cause the disease listeriosis in both humans and animals. For the epidemiological investigation of listeriosis detection and characterisation of the organism are important steps. Paper I. There are few reports on the incidence of L. monocytogenes in clinical samples from humans in India. Therefore, we investigated 144 samples from immunocompromised patients. L. monocytogenes was isolated from two placental bits from women with poor obstetric history, one patient with renal failure and three other patients. Five isolates were positive for the virulence genes hlyA, actA and iap. The sixth isolate was positive for hlyA and actA genes. Paper II. Characterisation of 601 human L. monocytogenes isolates causing invasive listeriosis during the period 1986 to 2007 in Sweden reveals a decrease in serovar 4b strains. Since 1996, serovar 1/2a has become the predominant serovar causing human listeriosis: PFGE analysis revealed two clusters including different serotypes suggesting that we need more studies on genetic relatedness among clinical isolates. Paper III. The incidence of Listeria species in seafood from markets in Goa was studied. One hundred and fifteen raw/fresh seafoods bought at the fish markets were sampled and tested for presence of Listeria spp. L. monocytogenes was detected in 10 samples. L. monocytogenes in raw seafood may pose a health risk in kitchen if contaminating ready-to-eat food. Paper IV. Gravad and cold-smoked salmon are associated with human listeriosis in Sweden. L. monocytogenes was isolated from 11 of 56 products. Serovar 1/2a was predominant, followed by 4b. REA/PFGE typing of the isolates identified five types of L. monocytogenes. One type was identical to a human type, two other were closely related.These findings suggest that gravad and cold-smoked salmon are still possible sources of listeriosis in Sweden. Paper V. Many outbreaks of listeriosis have been related to consumption of dairy-associated products. Therefore, 123 farm bulk milk samples in India and 20 cervico-vaginal samples from dairy cows with reproductive disorders were investigated for L. monocytogenes. L. monocytogenes was isolated from 17.9% of bulk milk samples and from 10% of cervico-vaginal swabs. The virulence gene hlyA was detected in all isolates. These findings represent a public health risk where unpasteurised milk and milk products are largely consumed. Paper VI. Isolates of L. monocytogenes (n=177) from 22 animal species were characterized and compared with human strains isolated between 1986-2006 in Sweden. Although many animals and humans shared pulsovars, they did not appear at the same time or with the same proportion of strains. The pulsovars shared by both animals and humans may indicate that there is an exchange of L. monocytogenes strains between these two groups due to either direct or indirect transmission. / <p>The work is done in cooperation with the School of Hospitality, Culinary Arts &amp; Meal Science, Örebro UniversityVishal Singh Parihar, Örebro University, Department of Restaurant and Culinary Arts, Sörälgsvägen 2, SE-712 60 Grythyttan, Sweden or ICAR Research Complex for Goa, Ela, Old Goa – 403402, Goa, India. Phone 0832-2284678/79; Fa:0832-2285649. E-mail: drvishu@yahoo.co.in</p>