Molecular characterization of the first leptospires isolated from goats in Brazil

Two Leptospira sp. isolates were obtained by the first time from goats in Brazil and characterized by sequencing rrs, rpoB and secY genes, PFGE and typing with monoclonal antibodies. Both isolates are identical and belong to Leptospira santarosai. Analysis of the rrs and the rpoB genes sequences revealed 100% identity between the goat isolates and the Bananal reference strain. When secY sequences of the two isolates were compared to each other, it was observed that they had identical sequences. However, when compared to that of the Bananal reference strain, there were 15 mismatches along the 549 bp secY sequence. In conclusion, molecular methods are increasingly useful for the characterization of leptospires and allowed to identify those isolates of caprine origin as closely related but not identical to serovar Bananal, and constitute a new type named Carioca.

Surveillance for leptospirosis in the Americas, 1996-2005: a review of data from ministries of health / Vigilancia de la leptospirosis en la Región de las Américas, de 1996 al 2005: análisis de los datos de los ministerios de salud

Objective. To characterize current leptospirosis reporting practices in the Americas.Methods. Information was collected from the official websites of national ministries ofhealth from the Americas region and two international organizations; personal communications;and three international morbidity databases. For all sources other than the morbiditydatabases, the review was limited to official reports citing clinically suspected and laboratoryconfirmed leptospirosis cases or deaths during the period 1996–2005.Results. A total of 73 out of 1 644 reports met the selection criteria and were included inthe analysis. Published leptospirosis data were available from half of the countries/sovereignterritories (24 out of 48), and 18 of them had mandatory notification policies for leptospirosis.The sum of the median number of leptospirosis cases notified annually by the 24 countries/territories was 4 713.5, but just three countries (Brazil, Costa Rica, and Cuba) accounted for83.1% (3 920 cases) of the notifications. Eight (16.7%) countries reported deaths due to leptospirosis.The sum of the median number of deaths reported annually for the eight countrieswas 380, but 349 (91.8%) were reported by Brazil.Conclusions. Notification practices in the Americas for leptospirosis are limited. Therefore,the numbers of cases and deaths reported are not representative for the region. The lack ofleptospirosis data for many countries/territories may reflect weaknesses in certain aspects ofnational surveillance systems, including mandatory reporting policies, clinical laboratory infrastructurefor performing case confirmation, and capacity to collect reported cases. Improvedsurveillance of leptospirosis cases and deaths in the Americas is needed to allow monitoring ofregional epidemiological patterns and to estimate the burden of this important disease.

Isolation of leptospira Serovars Canicola and Copenhageni from cattle urine in the state of Paraná, Brazil / Isolamento de leptospiras dos Sorovares Canicola e Copenhageni em amostras de urina de bovinos no estado do Paraná, Brasil

In 2001, 698 urine samples were randomly collected from cattle at a slaughterhouse in the State of Paraná, Brazil. Direct examination using dark field microscopy was carried out immediately after collection. Five putative positive samples were cultured in modified EMJH medium, yielding two positive cultures (LO-14 and LO-10). Typing with monoclonal antibodies revealed that the two isolates were similar to Canicola (LO-14) and Copenhageni (LO-10). Microscopic agglutination test results show that Hardjo is the most common serovar in cattle in Brazil. Rats and dogs are the common maintenance hosts of serovars Copenhageni and Canicola. The excretion of highly pathogenic serovars such as Copenhageni and Canicola by cattle can represent an increasing risk for severe leptospirosis is large populations, mainly living in rural areas.

No ano de 2001, 698 amostras de urina foram colhidas aleatoriamente, durante o abate em um frigorífico do Estado do Paraná, Brasil. O exame direto em microscópio de campo escuro foi realizado imediatamente após a colheita. As cinco amostras de urina positivas neste exame foram semeadas em meio EMJH modificado, sendo possível o crescimento de leptospiras em duas (LO-14 e LO-10). As estirpes isoladas foram tipificadas, por painel de anticorpos monoclonais, como mais similares ao perfil das amostras de referência dos sorovares Canicola (LO-14) e Copenhageni (LO-10). No Brasil, inquéritos sorológicos utilizando a prova de soroaglutinação microscópica mostram o predomínio de reações para o sorovar Hardjo em bovinos. Roedores e cães são os reservatórios mais comuns dos sorovares Copenhageni e Canicola, respectivamente. A eliminação dos sorovares Canicola e Copenhageni pela espécie bovina pode resultar em um aumento na ocorrência de casos graves de leptospirose humana, principalmente na população rural.

Isolation of leptospira santarosai, serovar guaricura from buffaloes (Bubalus bubalis) in Vale do Ribeira, São Paulo, Brazil

In April 1998 urine samples from adult female buffaloes were collected in a farm located in Registro, Vale do Ribeira, São Paulo State, Brazil. The urine samples obtained after furosemide injection were immediately transported to the laboratory in liquid modified EMJH medium and seeded, by the serial dilution technique, into Fletcher's or modified EMJH-0.2 (per cent) agar, both of them with 5-fluorouracil 100mg/mL. The intraperitoneoum inoculation of 0.5 mL was also performed with each urine sample in young, adult hamsters (Mesocricetus auratus). All samples seeded directly in culture medium were contaminated. The hamsters did not show any sign of disease and were killed at the 21st post inoculation day. At this time kidney cultures of these animals were performed and from one of them, one leptospira strain (M04-98) was isolated, identified as belonging to serogroup Sejroe by Microscopic Agglutination Test (MAT) with a panel of 36 rabbit sera against serovars representative for the pathogenic serogroups. Subsequently, MAT was carried out with antisera against the 19 reference strains of serogroup Sejroe, revealing a close relationship with serovar guaricura. Afterwards the MAT was done with a panel of 18 monoclonal antibodies representative for serovars of serogroup Sejroe. The histogram closely resembled that of serovar guaricura. So Cross Agglutination Absorption Test (CAAT) was carried out with the buffalo isolate and guaricura, supporting the relationship between the buffalo isolate and serovar guaricura.