Identification of a novel protein in the genome sequences of Leptospira interrogans with the ability to interact with host's components.

BACKGROUND: Leptospirosis is an infectious disease that affects humans and animals worldwide. The etiological agents of this disease are the pathogenic species of the genus Leptospira. The mechanisms involved in the leptospiral pathogenesis are not full understood. The elucidation of novel mediators of host-pathogen interaction is important in the detection of virulence factors involved in the pathogenesis of leptospirosis. OBJECTIVE: This work focused on identification and characterization of a hypothetical protein of Leptospira encoded by the gene LIC10920. METHODS: The protein of unknown function was predicted to be surface exposed. Therefore, the LIC10920 gene was cloned and the protein expressed in Escherichia coli BL21 (DE3) Star pLysS strain. The recombinant protein was purified by metal affinity chromatography and evaluated with leptospirosis human serum samples. The interaction with host components was also performed. RESULTS: The recombinant protein was recognized by antibodies present in leptopsirosis human serum, suggesting its expression during infection. Immunofluorescence and intact bacteria assays indicated that the bacterial protein is surface-exposed. The recombinant protein interacted with human laminin, in a dose-dependent and saturable manner and was named Lsa24.9, for Leptospiral surface adhesin, followed by its molecular mass. Lsa24.9 also binds plasminogen (PLG) in a dose-dependent and saturable fashion, fulfilling receptor ligand interaction. Moreover, Lsa24.9 has the ability to acquire PLG from normal human serum, exhibiting similar profile as observed with the human purified component. PLG bound Lsa24.9 was able of generating plasmin, which could increase the proteolytic power of the bacteria. CONCLUSIONS: This novel leptospiral protein may function as an adhesin at the colonization steps and may help the invasion process by plasmin generation at the bacterial cell surface.

Identification of a novel protein in the genome sequences of Leptospira interrogans with the ability to interact with host's components

Background Leptospirosis is an infectious disease that affects humans and animals worldwide. The etiological agents of this disease are the pathogenic species of the genus Leptospira. The mechanisms involved in the leptospiral pathogenesis are not full understood. The elucidation of novel mediators of host-pathogen interaction is important in the detection of virulence factors involved in the pathogenesis of leptospirosis. Objective This work focused on identification and characterization of a hypothetical protein of Leptospira encoded by the gene LIC10920. Methods The protein of unknown function was predicted to be surface exposed. Therefore, the LIC10920 gene was cloned and the protein expressed in Escherichia coli BL21 (DE3) Star pLysS strain. The recombinant protein was purified by metal affinity chromatography and evaluated with leptospirosis human serum samples. The interaction with host components was also performed. Results The recombinant protein was recognized by antibodies present in leptopsirosis human serum, suggesting its expression during infection. Immunofluorescence and intact bacteria assays indicated that the bacterial protein is surface-exposed. The recombinant protein interacted with human laminin, in a dose-dependent and saturable manner and was named Lsa24.9, for Leptospiral surface adhesin, followed by its molecular mass. Lsa24.9 also binds plasminogen (PLG) in a dose-dependent and saturable fashion, fulfilling receptor ligand interaction. Moreover, Lsa24.9 has the ability to acquire PLG from normal human serum, exhibiting similar profile as observed with the human purified component. PLG bound Lsa24.9 was able of generating plasmin, which could increase the proteolytic power of the bacteria. Conclusions This novel leptospiral protein may function as an adhesin at the colonization steps and may help the invasion process by plasmin generation at the bacterial cell surface.

Occurrence of anti-Leptospira antibodies in dogs in the southwestern region of the state of São Paulo / Ocorrência de anticorpos anti-Leptospira em cães na região sudeste do estado de São Paulo

Introduction: Leptospirosis is an emerging disease with different prevalence in dog populations. Dogs are crucial in the disease epidemiology, acting as accidental or maintenance hosts. Infective serovars present different geographic distribution among these populations, depending on exposure to hosts from infected wild or domestic animal reservoirs. The most common serovars that infect dogs prior to the introduction of the vaccines against leptospirosis were Icterohaemorrhagiae and Canicola. Objective: To analyze the occurrence of anti-leptospira antibodies in dogs from southwestern region of the state of São Paulo, using the microscopic agglutination test (MAT).(AU)

Occurrence of anti-Leptospira antibodies in dogs in the southwestern region of the state of São Paulo / Ocorrência de anticorpos anti-Leptospira em cães na região sudeste do estado de São Paulo

Introduction: Leptospirosis is an emerging disease with different prevalence in dog populations. Dogs are crucial in the disease epidemiology, acting as accidental or maintenance hosts. Infective serovars present different geographic distribution among these populations, depending on exposure to hosts from infected wild or domestic animal reservoirs. The most common serovars that infect dogs – prior to the introduction of the vaccines against leptospirosis – were Icterohaemorrhagiae and Canicola. Objective: To analyze the occurrence of anti-leptospira antibodies in dogs from southwestern region of the state of São Paulo, using the microscopic agglutination test (MAT).

Protective Immunity and Reduced Renal Colonization Induced by Vaccines Containing Recombinant Leptospira interrogans Outer Membrane Proteins and Flagellin Adjuvant.

Leptospirosis is a global zoonotic disease caused by different Leptospira species, such as Leptospira interrogans, that colonize the renal tubules of wild and domestic animals. Thus far, attempts to develop effective leptospirosis vaccines, both for humans and animals, have failed to induce immune responses capable of conferring protection and simultaneously preventing renal colonization. In this study, we evaluated the protective immunity induced by subunit vaccines containing seven different recombinant Leptospira interrogans outer membrane proteins, including the carboxy-terminal portion of the immunoglobulinlike protein A (LigA(C)) and six novel antigens, combined with aluminum hydroxide (alum) or Salmonella flagellin (FliC) as adjuvants. Hamsters vaccinated with the different formulations elicited high antigen-specific antibody titers. Immunization with LigA(C), either with alum or flagellin, conferred protective immunity but did not prevent renal colonization. Similarly, animals immunized with LigA(C) or LigA(C) coadministered with six leptospiral proteins with alum adjuvant conferred protection but did not reduce renal colonization. In contrast, immunizing animals with the pool of seven antigens in combination with flagellin conferred protection and significantly reduced renal colonization by the pathogen. The present study emphasizes the relevance of antigen composition and added adjuvant in the efficacy of antileptospirosis subunit vaccines and shows the complex relationship between immune responses and renal colonization by the pathogen.

Antibacterial activity and chemical compounds of leaves and branches of Protium hebetatum / Atividade anti bacteriana e compostos químicos das folhas e ramos de Protium hebetatum

ABSTRACT The extracts and fractions of leaves and branches of Protium hebetatum D. C. Daly (Burseraceae) were investigated for their antibacterial activity and chemical composition. The methanol extract of branches (EMG) was considered active against the Escherichia coli and the Proteus vulgaris, showing an inhibition zone of 13 mm, and was selected for bioassay-guided phytochemical fractionation. From the technique of broth microdilution, the extract was considered a moderate inhibitor against Staphylococcus aureus, Pseudomonas aeruginosa and Enterococcus faecalis, with a minimum inhibitory concentration (MIC) of 1 mg/mL. The dichloromethane fraction was considered a moderate inhibitor against S. aureus (MIC of 1 mg/mL) and a potent inhibitor against E. faecalis (MIC of 0.5 mg/mL). F1, F2, F5 and F6 from chromatographic column of dichloromethane fraction were considered moderate inhibitors against S. aureus (MIC of 1 mg/mL). Through analysis by a gas chromatography mass spectrometry, eighteen compounds were identified, from which thirteen (isoeugenol, p-vinylguaiacol, metoxyeugenol, coumarin, 5-hydroxy-scopoletin, 4,7-dihydroxy-6-metoxicromam-2-one, 4[(1E]-3-hydroxy-1-propenyl)-2-methoxyphenol, piperonal, scoparon, o-guaiacol, spathulenol, seringol and antiarol) are unprecedented in these species. We also identified the triterpenes α-amyrin and β-amyrin, the steroids stigmasterol and sitosterol and the coumarin scopoletin, which was closely linked to the antibacterial activity of the samples.

RESUMO Atividade antibacteriana e compostos químicos de folhas e galhos de Protium hebetatum. Extratos e frações de folhas e galhos de Protium hebetatum D. C. Daly (Burseraceae) foram investigados quanto sua atividade antibacteriana e composição química. O extrato metanólico dos galhos (EMG) foi considerado ativo contra Escherichia coli e Proteus vulgaris, apresentando um halo de inibição de 13 mm, sendo selecionado para um fracionamento fitoquímico biomonitorado. A partir da técnica de microdiluição em caldo o EMG foi considerado um inibidor moderado contra Staphylococcus aureus, Pseudomonas aeruginosa e Enterococcus faecalis, apresentando uma concentração inibitória mínima (CIM) de 1mg/mL. A fração diclorometânica foi considerada inibidora moderada contra S. aureus (CIM de 1 mg/mL) e inibidora potente contra E. faecalis (CIM de 0,5 mg/mL). F1, F2, F5 e F6 provenientes da fração diclorometânica foram consideradas inibidoras moderadas contra S. aureus (CIM de 1 mg/mL). Através da análise por cromatografia gasosa acoplada a espectrometria de massa, foram identificados dezoitos compostos, dos quais treze (isoeugenol, p-vinilguaiacol, metoxieugenol, cumarina, 5-hidroxi-escopoletina, 4,7-dihidroxi-6-metoxicromam-2-ona, 4[(1E]-3-hidroxi-1-propenil)-2-methoxifenol, piperonal, escoparona, o-guaiacol, espatulenol, seringol e antiarol) foram identificados pela primeira vez nesta espécie. Foram também identificados os triterpenos α-amirina e β-amirina, os esteroides estigmasterol e sitosterol e a cumarina escopoletina, que estão intimamente ligados à atividade antibacteriana da espécie.

Methods for thermal inactivation of pathogens in mozzarella: a comparison between stretching and pasteurization / Métodos para inativação térmica de patógenos em mozarela: comparação entre filagem e pasteurização

This study aimed to evaluate the efficiency of stretching in the reduction of pathogens when compared to milk pasteurization, the official method to ensure safe cheese production. Whole buffalo milk was contaminated with Mycobacterium fortuitum, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. Part of the milk was used in mozzarella production and the other part was submitted to holder pasteurization. Pathogens were quantified before and after thermal processing (mozzarella stretching and milk pasteurization). Pasteurization and stretching led to the following reductions in log cycles, respectively: 4.0 and 6.3 for Mycobacterium sp.; 6.0 and 8.4 for Listeria sp.; >6.8 and 4.5 for Staphylococcus sp.; and >8.2 and 7.5 for Salmonella sp.

Este estudo teve como objetivo avaliar a eficácia da filagem na redução de patógenos,em comparação coma pasteurizaçãodo leite, que é o método oficialpara garantir aprodução de queijos seguros. Leite de búfala integral foi contaminado com Mycobacterium fortuitum, Listeria monocytogenes, Salmonella typhimurium e Staphylococcus aureus. Parte desse leite foi empregada na fabricação da mozarela e outra parte foi submetida à pasteurização lenta. Os patógenosforam quantificadosantes e após os processos térmicos (filagem da mozarela e pasteurização do leite). As reduções, em ciclos logarítmicos, causadas pela pasteurização e pela filagem, respectivamente, foram: 4,0 e 6,3 de Mycobacterium sp., 6,0 e 8,4 de Listeria sp., >6,8 e 4,5 de Staphylococcus sp. e >8,2 e 7,5 de Salmonella sp.

Methods for thermal inactivation of pathogens in mozzarella: a comparison between stretching and pasteurization / Métodos para inativação térmica de patógenos em mozarela: comparação entre filagem e pasteurização

This study aimed to evaluate the efficiency of stretching in the reduction of pathogens when compared to milk pasteurization, the official method to ensure safe cheese production. Whole buffalo milk was contaminated with Mycobacterium fortuitum, Listeria monocytogenes, Salmonella typhimurium, and Staphylococcus aureus. Part of the milk was used in mozzarella production and the other part was submitted to holder pasteurization. Pathogens were quantified before and after thermal processing (mozzarella stretching and milk pasteurization). Pasteurization and stretching led to the following reductions in log cycles, respectively: 4.0 and 6.3 for Mycobacterium sp.; 6.0 and 8.4 for Listeria sp.; >6.8 and 4.5 for Staphylococcus sp.; and >8.2 and 7.5 for Salmonella sp.(AU)

Este estudo teve como objetivo avaliar a eficácia da filagem na redução de patógenos,em comparação coma pasteurizaçãodo leite, que é o método oficialpara garantir aprodução de queijos seguros. Leite de búfala integral foi contaminado com Mycobacterium fortuitum, Listeria monocytogenes, Salmonella typhimurium e Staphylococcus aureus. Parte desse leite foi empregada na fabricação da mozarela e outra parte foi submetida à pasteurização lenta. Os patógenosforam quantificadosantes e após os processos térmicos (filagem da mozarela e pasteurização do leite). As reduções, em ciclos logarítmicos, causadas pela pasteurização e pela filagem, respectivamente, foram: 4,0 e 6,3 de Mycobacterium sp., 6,0 e 8,4 de Listeria sp., >6,8 e 4,5 de Staphylococcus sp. e >8,2 e 7,5 de Salmonella sp.(AU)

Anticorpos anti-Leptospira spp. em ovinos do município de Monte Negro, estado de Rondônia / Anti-Leptospira spp. antibodies in ovines from Monte Negro municipality, Rondonia state, Brazil

O presente estudo determinou a prevalência de anticorpos anti-Leptospira spp. em ovinos do Município de Monte Negro, RO. Foram examinados soros de 141 ovinos de raça, idade e sexo variados provenientes de 15 fazendas, pela técnica de Soroaglutinação Microscópica. Doze (80,0%) propriedades apresentaram pelo menos um animal reagente. Títulos de anticorpos iguais ou superiores a 100 foram detectados em 47 (33,3%) animais, e os sorovares mais frequentes foram Patoc (29,7%), Autumnalis (14,8%), Pyrogenes (10,6%), Australis (4,2%), Bratislava (4,2%), Hardjo (4,2%), Icterohaemorrhagiae (4,2%), Castellonis (2,1%) e Hebdomadis (2,1%). Em 11 (23,4%) soros não foi possível a determinação do provável sorovar envolvido na reação. Alerta-se também para a possibilidade de infecção no homem, tendo em vista as características regionais de fronteira agrícola amazônica.

The present study determined the prevalence of anti-Leptospira spp.antibodies in 141 ovines from 15 farms of the Monte Negro Municipality, Rondonia State, Brazil, by the microscopic agglutination test. Twelve (80.0%) farms presented at least 1 reactive animal. Antibodies titers of ? 100 were detected in 47 (33.3%) animals, the most frequent serovars being Patoc (29.7%), Autumnalis (14.8%), Pyrogenes (10.6%), Australis (4.2%), Bratislava (4.2%), Hardjo (4.2%), Icterohaemorrhagiae (4.2%), Castellonis (2.1%) and Hebdomadis (2.1%). In 11 (23.4%) sera it was not possible to determine the most frequent serovar involved. The results raise a warning as to the possibility of infection in the human being by Leptospira in light of the regional characteristics of the Amazon agricultural frontier.

ANTICORPOS ANTI-LEPTOSPIRA SPP. EM OVINOS DO MUNICÍPIO DE MONTE NEGRO, ESTADO DE RONDÔNIA

ABSTRACT The present study determined the prevalence of anti-Leptospira spp.antibodies in 141 ovines from 15 farms of the Monte Negro Municipality, Rondonia State, Brazil, by the microscopic agglutination test. Twelve (80.0%) farms presented at least 1 reactive animal. Antibodies titers of ? 100 were detected in 47 (33.3%) animals, the most frequent serovars being Patoc (29.7%), Autumnalis (14.8%), Pyrogenes (10.6%), Australis (4.2%), Bratislava (4.2%), Hardjo (4.2%), Icterohaemorrhagiae (4.2%), Castellonis (2.1%) and Hebdomadis (2.1%). In 11 (23.4%) sera it was not possible to determine the most frequent serovar involved. The results raise a warning as to the possibility of infection in the human being by Leptospira in light of the regional characteristics of the Amazon agricultural frontier.

RESUMO O presente estudo determinou a prevalência de anticorpos anti-Leptospira spp. em ovinos do Município de Monte Negro, RO. Foram examinados soros de 141 ovinos de raça, idade e sexo variados provenientes de 15 fazendas, pela técnica de Soroaglutinação Microscópica. Doze (80,0%) propriedades apresentaram pelo menos um animal reagente. Títulos de anticorpos iguais ou superiores a 100 foram detectados em 47 (33,3%) animais, e os sorovares mais frequentes foram Patoc (29,7%), Autumnalis (14,8%), Pyrogenes (10,6%), Australis (4,2%), Bratislava (4,2%), Hardjo (4,2%), Icterohaemorrhagiae (4,2%), Castellonis (2,1%) e Hebdomadis (2,1%). Em 11 (23,4%) soros não foi possível a determinação do provável sorovar envolvido na reação. Alerta-se também para a possibilidade de infecção no homem, tendo em vista as características regionais de fronteira agrícola amazônica.

MILK BOILING EVALUATION ON INACTIVATING PATHOGENS IN COW, GOAT AND BUFFALO WHOLE MILK EXPERIMENTALLY CONTAMINATED / AVALIAÇÃO DA FERVURA NA INATIVAÇÃO DE PATÓGENOS EM LEITE INTEGRAL DE VACA, CABRA E BÚFALA EXPERIMENTALMENTE CONTAMINADOS

The inactivation of Staphylococcus aureus (ATCC 25923), Salmonella Typhimurium (ATCC 14028) and Mycobacterium fortuitum (NCTC 8573) inoculated in cow, goat and buffalo whole milk submitted to the boiling was evaluated. Milk samples, previously treated, were contaminated with the pathogens and boiled. Tree replicates were performed with each kind of milk. Samples were analyzed immediately before and after the boiling, and also after 24h under refrigeration. Samples were serial diluted and official method was used to quantify Staphylococcus aureus, and it was adapted to get the MPN of Salmonella. The number of CFU of Mycobacterium was obtained on Löwenstein-Jensen medium, incubated to 37°C/5 days. Before boiling the milk samples had from 6.8 to 8.0 log CFU or MPN/mL, of each agent. After boiling, Staphylococcus was recovery in one of three cow milk samples (1x10 CFU/mL), and Salmonella in all three buffalo milk samples (0.3 MPN/mL each). Mycobacterium was not detected in any of analysis made after the boiling. Boiled and refrigerated samples showed no growth in all of them. Under the studied conditions, the results prove the effectiveness of domestic boiling to reduce biological risk to minimum levels.

A fervura do leite foi avaliada quanto à inativação do Staphylococcus aureus (ATCC 25923), Salmonella Typhimurium (ATCC 14028) e Mycobacterium fortuitum (NCTC 8573) inoculados em leite integral de vaca, cabra e búfala. As amostras de leite, previamente tratadas, foram contaminadas com os patógenos. Três repetições foram realizadas com cada tipo de leite. As amostras foram analisadas antes e depois da fervura e também após 24h sob refrigeração. As amostras foram diluídas em série e a metodologia oficial foi usada para quantificar Staphylococcus aureus, e adaptada para se obter o NMP de Salmonella. O número de UFC de Mycobacterium foi obtido em meio Löwenstein-Jensen, incubados a 37°C/5 dias. Antes da fervura, as amostras tinham entre 6,8 e 8,0 log UFC ou NMP/mL, de cada agente. Após a fervura, Staphylococcus foi recuperado em uma das 3 amostras de leite de vaca (1x10 UFC/mL), e Salmonella em todas as três amostras de leite de búfala (0,3 NMP/mL cada). Mycobacterium não foi detectado em nenhuma das analises realizadas após a fervura. As amostras fervidas e refrigeradas mostraram ausência de crescimento de todos os agentes. Nas condições estudadas, os resultados comprovam a efetividade da fervura do leite em reduzir o risco microbiológico a níveis mínimos. PALAVRAS-CHAVE: Microbiologia. Mycobacterium fortuitum. Salmonella Typhimurium. Staphylococcus aureus. Tratamento térmico.

Avaliação da fervura na inativação de patógenos em leite integral de vaca, cabra e búfala experimentalmente contaminados / Milk boiling evaluation on inactivating pathogens in cow, goat and buffalo whole milk experimentally contaminated

A fervura do leite foi avaliada quanto à inativação de Staphylococcus aureus (ATCC 25923), Salmonella Typhimurium (ATCC 14028) e Mycobacterium fortuitum (NCTC 8573) inoculados em leite integral de vaca, cabra e búfala. As amostras de leite, previamente tratadas, foram contaminadas com os patógenos. Três repetições foram realizadas com cada tipo de leite. As amostras foram analisadas antes e depois da fervura e também após 24h sob refrigeração. As amostras foram diluídas em série e a metodologia oficial foi usada para quantificar Staphylococcus aureus, e adaptada para se obter o NMP de Salmonella. O número de UFC de Mycobacterium foi obtido em meio Löwenstein-Jensen, incubados a 37°C/5 dias. Antes da fervura, as amostras tinham entre 6,8 e 8,0 log UFC ou NMP/mL, de cada agente. Após a fervura, Staphylococcus foi recuperado em uma das 3 amostras de leite de vaca (1x10 UFC/mL), e Salmonella em todas as três amostras de leite de búfala (0,3 NMP/mL cada). Mycobacterium não foi detectado em nenhuma das analises realizadas após a fervura. As amostras fervidas e refrigeradas mostraram ausência de crescimento de todos os agentes. Nas condições estudadas, os resultados comprovam a efetividade da fervura do leite em reduzir o risco microbiológico a níveis mínimos.

The inactivation of Staphylococcus aureus (ATCC 25923), Salmonella Typhimurium (ATCC 14028) and Mycobacterium fortuitum (NCTC 8573) inoculated in cow, goat and buffalo whole milk submitted to the boiling was evaluated. Milk samples, previously treated, were contaminated with the pathogens and boiled. Tree replicates were performed with each kind of milk. Samples were analyzed immediately before and after the boiling, and also after 24h under refrigeration. Samples were serial diluted and official method was used to quantify Staphylococcus aureus, and it was adapted to get the MPN of Salmonella. The number of CFU of Mycobacterium was obtained on Löwenstein-Jensen medium, incubated to 37°C/5 days. Before boiling the milk samples had from 6.8 to 8.0 log CFU or MPN/mL, of each agent. After boiling, Staphylococcus was recovery in one of three cow milk samples (1x10 CFU/mL), and Salmonella in all three buffalo milk samples (0.3 MPN/mL each). Mycobacterium was not detected in any of analysis made after the boiling. Boiled and refrigerated samples showed no growth in all of them. Under the studied conditions, the results prove the effectiveness of domestic boiling to reduce biological risk to minimum levels.

Soroprevalência de leptospirose em fêmeas bovinas em idade reprodutiva no estado da bahia

ABSTRACT The aim of this work was to determine the seroprevalence of leptospirosis in reproductive-age bovine females in Bahia State, Northeastern Brazil. The sampling was delineated for the determination of the prevalence of seropositive animals as well as herds positive for bovine leptospiroses (foci). The state was divided into 4 regions or sampling strata in which 10,823 bovine females aged 24 months allocated in 1,414 herds were sampled. The microscopic agglutination test (MAT), using 23 Leptospira spp. serovars as antigens, was employed as a diagnostic test. The herd was considered positive if at least one animal was seropositive. The prevalences of positive herds and seropositive animals in the state were 77.93% [75.73%79.99%] and 45.42% [42.00%48.88%], respectively. Serovar Hardjo (Hardjoprajitno) was the most frequent, with 34.49% [31.93%37.14%] of positive herds and 14.95% [12.59%17.67%] of seropositive animals in the different regions.

RESUMO O objetivo do presente trabalho foi determinar a soroprevalência da leptospirose em fêmeas bovinas em idade reprodutiva no Estado da Bahia. A amostragem foi delineada para a determinação da prevalência de propriedades positivas (focos) e de animais soropositivos para a leptospirose. O Estado foi dividido em quatro regiões ou estratos amostrais, nos quais foram examinadas 10.823 fêmeas bovinas com idade 24 meses distribuídas em 1.414 propriedades. A reação de Soroaglutinação Microscópica (SAM), empregando 23 sorovares de Leptospira spp. como antígenos, foi utilizada como teste diagnóstico. O rebanho foi considerado foco quando apresentou pelo menos um animal soropositivo. As prevalências de foco e de animais soropositivos no Estado foram de 77,93% [IC 95% = 75,73% 79.99%] e 45,42% [IC 95% = 42,00% 48,88%], respectivamente.

Soroprevalência da leptospirose em fêmeas bovinas em idade reprodutiva no estado de são paulo, brasil

ABSTRACT The objective of the present study was to determine the seroprevalence of bovine leptopirosis in São Paulo State, stratified in seven cattle production regions. It was based on the statistic delineation, serological samples and responses to the survey employed in the National Program for Control and Eradication of Brucelosis and Tuberculosis established by Ministry of Agriculture (2001). From the herds selected, serological analysis was only conducted on the cows 24 months old (excluding the males). The study took into consideration the herd size, the type of productive exploration, the reproductive handling, bovine practices and the commercialization system. The microscopic agglutination test (MAT) was applied on 8,216 serum samples from 1,021 different farms. The results showed that leptospirose infection occurs all over the seven regions of São Paulo State with 49.4% (CI 95% = 44.454.4%) animal seroprevalence and in 718 (71.3%; CI 95% = 68.574.0%) of the herds analyzed. Hardjo (46%) was the prevalent serovar for all the animals examined, followed by the Hardjo/Wolffi association (21%), Shermani (8.9%), Autumnalis (4.4%) and Grippotyphosa (3.9%). Leptospira spp. is present in all regions of the State of São Paulo and its occurrence is independent of the handling conditions and reproductive practices adopted in the herds.

RESUMO O presente estudo teve como objetivo determinar a soroprevalência da leptospirose bovina em fêmeas em idade reprodutiva do Estado de São Paulo, estratificado em sete regiões produtoras. Foram utilizados o delineamento estatístico, as amostras sorológicas e as informações contidas nos questionários empregados no Programa Nacional de Controle e Erradicação da Brucelose e Tuberculose (PNCETB), instituído pelo Ministério da Agricultura, Pecuária e Abastecimento, considerando-se a utilização de fêmeas bovinas com idade a 24 meses (excluindo-se machos), diferentes tipos de produção, práticas de manejo, finalidades de reprodução, tamanho dos rebanhos e sistema de comercialização. Realizou-se a Soroaglutinação Microscópica (SAM) em 8.216 amostras sorológicas de animais provenientes de 1.021 propriedades. De acordo com os resultados obtidos, a infecção por Leptospira spp. ocorre em todo o Estado de São Paulo, com soroprevalência de 49,4% (IC 95% = 44,4%-54,4%) nas fêmeas bovinas em idade reprodutiva e em 718 (71,3%; IC 95% = 68,5%-74%) das propriedades analisadas. O sorovar Hardjo (46%) e sua associação com o sorovar Wolffi (21%) foram prevalentes entre o total de animais sororeagentes, seguidos pelos sorovares Shermani (8,9%), Autumnalis (4,4%) e Grippotyphosa (3,9%). Leptospira spp. está distribuída por todo estado e independe do tipo de exploração, manejo e das práticas de reprodução adotadas nos rebanhos.

Frequency of anti-leptospires agglutinins in sows from a swine herd in the ibiúna municipality, state of são paulo, brazil

ABSTRACT A serologic survey was conducted among 164 sows from a single farrow-to-finish swine herd located in the Ibiúna municipality, state of São Paulo, Brasil, to determine the frequency of antileptospires agglutinins. For detection of anti-leptospires antibodies, the microscopic serumagglutination test (MAT) was carried out using live cultures of 22 pathogenic and two saprophytic Leptospira spp. serovars. The most frequent serovar was found crossing the results of frequency and titer of agglutinins, and sera presenting equal titers for two or more serovars were not considered for this analysis. Of the 164 sows, 27 (16.5%) were seropositive for at least one Leptospira spp. serovar. The most frequent serovar was Hardjo (Hardjobovis), with 13 (54.2%) reactant sera. Other reactant serovars and respective frequency were: Shermani (16.6%), Bratislava (12.5%), Autumnalis (12.5%) and Icterohaemorrhagiae (4.2%).

RESUMO Com o objetivo de determinar a freqüência de aglutininas anti-leptospiras, foi realizado um inquérito sorológico em 164 matrizes de uma granja suína de ciclo completo localizada no Município de Ibiúna, Estado de São Paulo, Brasil. Para a detecção de anticorpos anti-leptospiras, foi utilizada a técnica de soroaglutinação microscópica (SAM), utilizando-se culturas vivas de 22 sorovares patogênicos e dois sorovares saprófitos de Leptospira spp. Para a determinação do sorovar mais provável, foram considerados o título de aglutininas e a freqüência de soros reagentes, e soros que apresentaram títulos iguais para dois ou mais sorovares foram excluídos desta análise. Das 164 matrizes suínas, 27 (16,5%) foram soropositivas para pelo menos um dos sorovares empregados. O sorovar mais provável foi o Hardjo (Hardjobovis), com 13 (54,2%) soros reagentes. Também foram constatadas reações sorológicas para os seguintes sorovares: Shermani (16,6%), Bratislava (12,5%), Autumnalis (12,5%) and Icterohaemorrhagiae (4,2%).

Utilização de uma técnica rápida para o diagnóstico de mycobacterium bovis em amostras de leite experimentalmente inoculadas

ABSTRACT The modified Middlebrook 7H11 cultivation technique was comparedwith the tradicional culture techniqueby using the Stonebrink medium. The purpose of this comparison was to to evaluate the sensitivity and the time for the detection of positive cultures of mycobacteria in milk samples experimentally inoculated with Mycobacterium bovis (strain AN5), at a dilution of 10-2 , and submitted to two types of procedures: technique 1 (skin milk) and technique 2 (pellet), decontamined by modified the Petroff´s method, added with Tween 80, and compared with total milk samples submitted to tradicional Petroff method in the same media types. The results of the two tecniques were compared with each other by the non-parametric Wilcoxon test and Mann-Whitney. The results obtained from this experiment showed that: techniques 1 and 2 produced great number of colonies and higher proportion of positive culture than the tradicional one by using total milk; the time needed for the detection of mycobacteria colonies was slightly shorter by the thin layer technique than by the tradicional culture method; in order to improve the epidemiological surveillance, this technique should be used as a complementary method to the tradicional one in the diagnosis of bovine tuberculosis.

RESUMO O meio de Middlebrook 7H11 modificado foi comparado ao meio de Stonebrink, a fim de se avaliar a sensibilidade e o tempo de detecção de micobactérias em amostras de leite, experimentalmente inoculadas com Mycobacterium bovis (estirpe AN5), em uma diluição 10-2, e submetidas a duas diferentes técnicas de processamento: gordura (técnica 1) e sedimento (técnica 2), descontaminadas pelo método de Petroff modificado (adicionado de Tween 80), confrontadas com a técnica do leite total submetida ao método de Petroff tradicional. Os resultados destas técnicas (1 e 2) foram comparados entre si pelos testes não paramétricos de Wilcoxon e de MannWhitney e demonstraram que as técnicas 1 e 2 forneceram maior recuperação de micobactérias e proporção de cultivos positivos nos meios de Stonebrink e Middlebrook 7H11; o meio de Middlebrook 7H11 permitiu a visualização precoce das micobactérias, podendo ser utilizado como uma técnica de diagnóstico rápida da tuberculose bovina, em amostras de leite, para fins de vigilância epidemiológica.

Aneurisma da arteria esplenica. / Aneurysm of splenic artery

Apresentam os autores a experiencia de dois casos de aneurismas de arteria esplenica agudos no servico de cirurgia do HECC. Fazem revisao da literatura enfatizando o exame fisico e radiologico simples do abdomen dados fundamentais para o diagnostico em que o sinal do "anel de sinete" devera ser sempre pesquisado. Referem ainda os magnificos resultados obtidos com a aneurismectomia e esplenectomia