A new non-species dependent ELISA for detection of antibodies to Borrelia burgdorferi s. l. in zoo animals.

Although Lyme borreliosis is regarded as one of the most important zoonotic diseases, in Europe very few research reports have documented examinations on free-ranging wild animal populations and no study on zoo animals has been published. One of the problems regarding the diagnosis of Lyme borreliosis in wild animals is often the lack of species specific secondary antibodies for serological tests. For our study on exposure of zoo animals to Borrelia (B.) burgdorferi s. l. and the occurrence of Lyme borreliosis in various German zoos, a non-species dependent ELISA was developed. Specific IgG antibodies against B. burgdorferi s. l. were detected by peroxidase labeled protein A or protein G conjugates. For this purpose, both conjugates were tested for their binding affinities to 160 different wild animal species representing 25 families and 7 orders. With 2 exceptions, all tested species reacted with either protein A or protein G, and 47 species reacted with both conjugates. In combination with an easy method for the long-term preservation of antigen-coated microtiter plates, the ELISA developed for this study could essentially facilitate serological examinations regarding Lyme borreliosis in wild animal sera. In summary, the results indicate commercially available protein A and protein G conjugates as useful alternatives to species specific secondary antibodies in various diagnostic assays on sera of a wide range of wild mammals. Therefore, this should be considered more often as versatile diagnostic tools in wildlife studies.

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.