The serological response of calves to Leptospira interrogans serovar hardjo vaccines and infection as measured by the microscopic agglutination test and anti-IgM and anti-IgG enzyme-linked immunosorbent assay

The microscopic agglutination test (MAT) and the anti-IgM and anti-IgG enzyme-linked immunosorbent assays (ELISA) were used to examine sera taken over the course of 16 weeks from 35 calves vaccinated and/or infected with Leptospira interrogans serovar hardjo. The relationship between the IgM and IgG responses to vaccination and infection were determined. The rapid and high rise in IgM levels following challenge made the anti-IgM ELISA a potentially good indicator of recently established infection although some transitory high levels were seen where infection did not become established. The slow IgG response to infection made the anti-IgG ELISA of limited diagnostic use.     

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.     

Problems associated with the serological diagnosis of Leptospira interrogans serovar hardjo infection in bovine populations

Data combining sequential bacteriology and serology from a longitudinal study of a dairy herd were used to demonstrate the limitations of serology as a diagnostic method in cross-sectional sampling of bovine populations. Whole-herd point serological prevalences showed considerable variation over a two-year sampling period (38.8 to 76.2 per cent), and this was mainly due to varying age-specific prevalence. Owing to the rapid decline in titres and the varying persistence of infection, point serological prevalences failed to approximate to cumulative infection rates (either past or present) at different times of the year. A higher estimate of the number of susceptible animals in the herd than is the case results in inaccurate information on true incidence rates and can confuse assessments of the susceptibility of different age groups, especially if only small numbers are sampled. A sampling exercise demonstrated that a 10-cow sample usually provided little useful information other than establishing the presence or absence of hardjo in the herd. Increasing the sample size markedly improved epidemiological information, investigations of clinical disease, assessments of vaccination needs and public health tracebacks. Preferably 10 sera from each of the yearling, first calver, second calver and older age groups should be tested. Serology was an inadequate indicator of infection in individual animals. Group geometric mean titres taken from a mean serological response curve were shown to have limited application in the interpretation of field data, unless infection had occurred in the previous two months.     

Experimental Leptospira borgpetersenii serovar hardjo infection of pregnant cattle

Objective To observe the effect upon the foetus of experimental infection of pregnant cattle with Leptospira borg-petersenii serovar hardjo .
Design A disease transmission study using pregnant cattle.
Procedure Fourteen heifers serologically negative to L hardjo were artificially inseminated and later challenged with a north-Queensland isolate of L hardjo by conjunctival inoculation. The heifers were serologically monitored and their urine examined for the presence of leptospires using culture and fluorescent-antibody tests at appropriate intervals. Elective caesarean sections were performed on pregnant heifers at 6.5 weeks after the challenge. Foetuses were examined using serological, histopathological, microbiological and fluorescent-antibody tests.
Results Ten of the heifers became pregnant, but three subsequently aborted before challenge. After challenge, all 14 heifers seroconverted and L hardjo was isolated from the urine of 6 of the 7 pregnant heifers. No evidence of foetal L hardjo infection was detected. Two of the foetuses had histopatho-logical lesions consistent with Neospora s p infection.
Conclusion It is likely that the isolate of L hardjo used in this study does not normally infect the foetus. Neospora s p may be a more significant cause of bovine reproductive wastage.     

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.     

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.     

Excretion of Leptospira interrogans serovar hardjo following calving or abortion

Leptospira interrogans serovar hardjo was demonstrated in the vaginal discharges of 12 experimentally infected heifers for up to eight days after abortion or calving. Organisms were recovered from the oviducts of heifers examined at slaughter eight to 22 days after calving or 32 to 91 days after infection. They were also recovered from the uteri of four heifers eight to 22 days after calving.     

Dihydrostreptomycin treatment of bovine carriers of Leptospira interrogans serovar hardjo

Ten non-pregnant heifers, experimentally infected with Leptospira interrogans serovar hardjo, were each given an intramuscular injection of dihydrostreptomycin at a dose rate of 25 mg kg-1. In five of these animals this treatment was repeated after 14 days. Treatment failed to remove established genital or renal infection in seven of the 10 heifers, although the number of organisms persisting was apparently reduced in these animals. Serovar hardjo infection persisted for at least 83 days in the oviduct and uterus and these tissues may be as important carrier sites for serovar hardjo as bovine kidney tissue.     

Leptospira interrogans serovar hardjo associated with bovine abortion in South Africa

Leptospira interrogans serovar hardjo was isolated from urine from dairy cattle in the Onderstepoort area. This was the first successful isolation of this serovar as sole agent causing an abortion storm in the Republic of South Africa. Abortions occurred as early as at 4 months' gestation.     

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.     

Restriction endonuclease analysis of Leptospira interrogans serovar hardjo isolates from cattle

The genomes of 253 strains of Leptospira interrogans serovar hardjo were examined by restriction endonuclease analysis of their DNA. The strains had been isolated from cattle at an abattoir (190), milk of agalactic cows (seven) and from aborted bovine fetuses (56). Two distinct genome types, Hardjoprajitno and Hardjobovis, were detected. The majority (91 per cent) of isolates from abattoir cattle were of the Hardjobovis type while most (76 per cent) of the isolates from clinical/pathological material were similar to Hardjoprajitno.