Comparative serological study of Leptospira serovar hardjo genotypes for use in the microscopic agglutination test.

A comparative serological study was conducted using the Leptospira microscopic agglutination test (MAT). Genotypes hardjoprajitno (HP), hardjo-bovis A (HA), and hardjo-bovis B (HB) were compared to determine which best detects hardjo antibody in cattle serum. A total of 2,431 cattle sera were tested. Sera were collected from 4 geographic regions of the United States. Samples were obtained without knowledge of breed, age, vaccination history, or herd health status. Of the sera collected, 60.7% (1,475) were negative at the 1 : 100 dilution for all three genotypes. Serological reactivity at the 1 : 100 dilution was identified in 956 (39.3%) of the sera tested. Considering the 956 positive sera, 941 (98.4%) reacted to HP, whereas the remaining 15 sera (1.6%) reacted to only HA and/or HB. A total of 394/941 (41.9%) HP positive sera failed to react to HA or HB. The results of this study support the conclusion that HP antigen was most sensitive in detection of hardjo antibody.     

牛、羊布鲁氏菌病试管凝集试验与微量凝集试验检测方法的比较

血清学检测方法因其快速、安全、高效等特点被广泛应用于布鲁氏菌病的检测和诊断。本文通过对保存的226份牛、羊血清样品同时使用试管凝集法与微量凝集试验进行检测并比较分析其在诊断中的意义。结果显示,微量法与试管法两者的符合率为97.79%,两种检测方法无统计学差异。但微量法的阳性检出率要高于试管法,同时由于微量法具有操作简便、快捷及结果判定更为直观、简单等优点,因此,微量凝集试验更适宜在我国基层布鲁氏菌病大量样本监测、诊断中推广应用。     

Control of Leptospira hardjo infection in beef cattle by whole-herd vaccination.

A whole-herd vaccination programme to control Leptospira hardjo infection was applied to a closed herd of approximately 800 beef cattle on the island of Luing in Scotland. An experimental vaccine was produced and the herd was vaccinated annually for five years. Progress was monitored by means of a catalytic model using data for age-specific serological prevalences and geometric mean titres. Any cattle introduced to the herd were subject to antibiotic treatment and quarantine, and at the end of the trial the whole herd was treated prophylactically with antibiotics to minimise the risk of residual infection. There was a progressive right shift in age-specific serological prevalences, and by the end of the trial all young stock entering the breeding herd were seronegative. The age-specific geometric mean titres demonstrated the cessation of an endemic cycle of hardjo infection in the herd. Birth cohort analysis supported the serological evidence of a high level of control, and bacteriological monitoring at the end of the trial indicated that hardjo had been eliminated from the herd.     

Production and characterization of monoclonal antibodies specific for Leptospira borgpetersenii serovar hardjo type hardjobovis and Leptospira interrogans serovar hardjo type hardjoprajitno.

Murine monoclonal antibodies were produced by immunizing BALB/c mice with a killed whole-cell antigen prepared from Leptospira borgpetersenii serovar hardjo type hardjobovis. Six of these antibodies recognized epitopes on the homologous antigen and on whole-cell antigen prepared from Leptospira interrogans serovar hardjo type hardjoprajitno. These antibodies did not cross-react with whole-cell antigens prepared from L. borgpetersenii serovar sejroe, 10 other pathogenic Leptospira serovars, or the saprophytic Leptospira biflexa serovar patoc. Three other monoclonal antibodies reacted with antigens prepared from the 2 hardjo serovars and serovar sejroe but not with antigens from the 10 other pathogenic serovars, or serovar patoc. The epitopes recognized by all of the hardjo-specific antibodies and 2 of the 3 hardjo/sejroe-specific antibodies were susceptible to sodium meta-periodate oxidation. All of the antibodies were characterized by Western blots with the hardjobovis whole-cell antigen. Each of the 9 monoclonal antibodies was inhibited from binding to the hardjobovis antigen by bovine sera which were obtained from cattle experimentally infected with hardjobovis and from field cattle, with anti-serovar hardjo microscopic agglutination test antibody titres ranging from 100 to 12800. Some of these antibodies may be suitable for incorporation into competitive enzyme immunoassays for the specific detection of antibodies to either of the hardjo serovars.     

Suppression of sheep and goat lymphocyte proliferation by sheep, goat, and sheep x goat hybrid trophoblast tissue cultures.

Immunosuppressive activity of conditioned medium from cultured ovine, caprine, and hybrid trophoblast tissue was examined. Conceptuses were obtained from naturally mated donor ewes and does at d 20 of gestation and trophoblast tissue was cultured for 24 h in medium supplemented with 15% calf serum and 1% antibiotic/antimycotic. Conditioned medium was added to pokeweed mitogen-stimulated sheep and goat lymphocyte cultures. Quantification of [3H]thymidine uptake by cells was used to measure lymphocyte proliferation. Ovine, caprine, and hybrid conditioned medium effectively suppressed sheep and goat lymphocyte proliferation (P less than .01). There were no differences (P greater than .05) between the immunosuppressive activity of the three tissue types on either sheep or goat lymphocytes. For all treatment groups, sheep lymphocytes were suppressed more than goat lymphocytes (P less than .05). These results indicate that, at d 20 of gestation, sheep x goat hybrid trophoblast tissue is capable of suppressing pokeweed mitogen-stimulated lymphocyte proliferation.     

Addition of rabbit serum to EMJH medium improves isolation of Leptospira interrogans serovar hardjo

The addition of 2% pooled rabbit serum to semi-solid commercial EMJH medium with EMJH enrichment and 0,5 mg of 5-fluorouracil per ml was found to enhance the growth rate and success of isolation of Leptospira interrogans serovar hardjo from bovine urine. Cultures made on media without serum had to be kept for more than 130 days before being discarded as negative.     

Diagnosis of Leptospira serovar hardjo Infection in Cattle in Canada

The investigations described were designed to identify the cause of serological reactions to Leptospira interrogans serovars hardjo and sejroe in Canadian cattle, and to confirm by culture a diagnosis of leptospirosis in cases of reproductive failure and atypical mastitis.     

Isolation of Leptospira hardjo from kidneys of Ontario cattle at slaughter.

Kidneys from 117 cattle from 110 Ontario farms were examined at slaughter for leptospires. Leptospira hardjo (hardjo-bovis A) was isolated from 11 kidneys and L. kennewicki from one. The isolations were all made (12/89, 13.5%) from beef cattle from feedlots, no isolates being obtained from dairy or beef cattle from extensive farms (0/28). Isolations were only made from cattle with antibody titers (greater than or equal to 20) against the serovars recovered. Isolation was more sensitive than immunofluorescence in identifying leptospira, particularly in animals with low antibody titers against L. hardjo. Leptospira were isolated from two kidneys with multiple gross lesions of focal nephritis, but there was no correlation between the presence of scanty kidney lesions and isolations of leptospira. Leptospira hardjo infection appears to be common in Ontario feedlot cattle.     

Antimicrobial drug susceptibility of Leptospira interrogans serovar hardjo isolated from cattle.

The susceptibility to commonly used drugs of 18 isolates of Leptospira hardjo from the kidneys of feedlot cattle from different sources was determined quantitatively. All isolates were susceptible to penicillin G, ampicillin, tetracycline, erythromycin and streptomycin. Susceptibility to sulphamethazine was ambiguous. No drug resistance was detected and the results were similar to those described for other serovars.     

Leptospira borgpetersenii serovar hardjo type hardjobovis in bovine embryos fertilized in vitro.

The association of Leptospira borgpetersenii serovar hardjo type hardjobovis with bovine embryos produced by in vitro fertilization was examined by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Morula stage embryos with an intact zona pellucida (ZP) were exposed to this spirochete for 24 h in culture medium, washed by the standard washing procedure as recommended by the International Embryo Transfer Society, and then examined. SEM showed typical helicoid leptospires on the surface and in the pores of the ZP. TEM showed cross and longitudinal sections of leptospires in the matrix and channels of the ZP, in the perivitelline and intercellular spaces, on the vitellus and in the embryonic cells. Some of the embryos that were penetrated showed damage to the membranes and the cytoplasm. The ineffectiveness of the washing procedure, for the removal of hardjobovis from exposed embryos may be of importance to the industry.