Epidemiology of Yersinia pseudotuberculosis and Y. enterocolitica infections in sheep in Australia.

Infections with Yersinia pseudotuberculosis serotype III and Y. enterocolitica serotype O2,3 were found to be common in Australian sheep flocks. Transmission of Y. pseudotuberculosis occurred in late winter and early spring, while Y. enterocolitica transmission occurred from midwinter to early summer. Excretion of Y. pseudotuberculosis was limited to the winter and spring period and was particularly common in 1- and 2-year-old sheep. Infection persisted for up to 14 weeks. Y. pseudotuberculosis infection did not confer immunity to natural infection with Y. enterocolitica. Y. enterocolitica excretion occurred year-round, with the greatest prevalence being in summer and autumn. Infection persisted for up to 29 weeks. Sheep less than 1 year old were most commonly infected with Y. enterocolitica. Infection with either Y. pseudotuberculosis or Y. enterocolitica was rare in aged sheep. Restriction endonuclease analysis of Y. pseudotuberculosis serotype III from sheep, cattle, deer, and pigs showed that the bacterial isolates were genetically indistinguishable. Similarly, Y. enterocolitica isolates from sheep were indistinguishable from those isolated from goats and cattle.

Genetic diversity among Australian and New Zealand isolates of Leptospira interrogans serovar pomona.

Restriction endonuclease analysis of 16 Australian and 4 New Zealand isolates of Leptospira interrogans serovar pomona showed that they could be divided into 3 genetic groups. Most of the isolates closely resembled the serovar kennewicki reference strain, and they all differed from the reference strain of serovar pomona. Based on these findings, it is suggested that vaccine manufacturers re-evaluate their choice of serovar pomona vaccine strain.

Comparison of the Johne's absorbed EIA and the complement-fixation test for the diagnosis of Johne's disease in cattle.

A commercially available absorbed ELISA for the diagnosis of Johne's disease (JD) (paratuberculosis) in cattle, the Johne's Absorbed EIA, was compared with the conventional complement-fixation test (CFT) used in Australia. Stored plasma from 3 Victorian dairy herds with a history of JD, sera from specimens submitted from animals showing clinical signs of JD and sera from the US National Repository for Paratuberculosis Specimens were used to determine the sensitivity of each test. The EIA detected 48.8% of 43 Australian animals with subclinical JD, while the CFT detected only 12 (21.4%) of 56 subclinically affected cattle. Of 150 subclinically infected US cattle, the EIA detected 47.3% and the CFT detected 52.0%. The EIA detected 59.7% of animals which at the time of sampling were shedding Mycobacterium paratuberculosis in their faeces, but showed no clinical signs of JD, while the CFT detected 57.3%. The EIA correctly identified 88.2% of 136 histologically confirmed clinical cases, and the CFT detected 83.4%. The specificity of each test was determined by testing sera collected at slaughter from animals residing in a known JD-free area of Australia, and from samples from the US National Repository of Paratuberculosis Specimens collected from certified-free herds in Wisconsin. The EIA was found to have a specificity of 99.8% when 998 Australian animals were used as the test population, and 99.0% when 196 US animals were used. The specificity of the CFT using Australian samples was 96.9% and 95.2% using American samples.

Leptospira interrogans serovar hardjo is not a major cause of bovine abortion in Victoria.

The aim of this study was to determine whether evidence could be obtained of foetal infection with Leptospira interrogans serovar hardjo in aborted foetuses collected from dairy farms. Material from 197 abortions occurring over a wide area of Victoria was collected over 3 years. None of 195 foetal kidney cultures or 7 cultures from membranes was positive for leptospiral organisms. Immunogold silver staining for leptospires was performed on sections of kidneys, lungs or heart from 156 foetuses, with negative results. Evidence of transient leptospiral infection in 11 of 123 foetuses was obtained by foetal heart blood serology. Two isolates of L. interrogans serovar hardjo were obtained from the urine of milking cows. These strains were examined by restriction endonuclease analysis and both were shown to be of the genotype Hardjobovis, as have been all Australian isolates studied so far. It appears that foetal infection with serovar hardjo is not associated with any substantial proportion of bovine abortions in Victoria, in contrast to the situation in Northern Ireland. The apparent absence from Victoria of the pathogenic genotype Hardjoprajitno is a possible explanation.

Bovine leptospirosis: microbiological and histological findings in cattle at slaughter.

Kidneys from cattle at slaughter were examined for the presence of leptospires. Of 218 (8.3%) kidneys leptospires were isolated from 18; all were identified as Leptospira interrogans serovar hardjo. None of the leptospire-infected kidneys had histopathological lesions indicative of leptospirosis and leptospires were demonstrated in only 2 by immunogold silver staining. Leptospires infected kidneys remained viable for at least 21 days when stored at 4 degrees but became non-viable within 14 days when stored frozen at -15 degrees.

Identification of Mortierella wolfii, a causative agent of mycotic abortion in cattle.

Of several media tested for their ability to induce asexual reproduction in confirmed or suspected isolates of Mortierella wolfii obtained from cattle in various geographical locations, only silage extract agar worked consistently and rapidly. Its use should provide a simple reliable culturing procedure to assist with identification of clinical isolates of this fungus. Temperature growth response curves and electrophoretic patterns of soluble protein extracts of these isolates with characteristic morphological features of M. wolfii indicate substantial variation amongst them.

Immunogold silver staining for visualization of leptospires in histologic sections.

An immunogold silver stain for leptospires in sections of Formalin-fixed, paraffin-embedded tissues is described. Leptospires were intensely stained, and nonspecific staining of the tissue background was negligible.

Immunofluorescent staining of leptospires in pepsin treated histologic sections.

A standard immunofluorescent method was modified for the staining of leptospires in formalin fixed, paraffin embedded tissues. Routine histologic sections were deparaffinized and treated with pepsin prior to staining. Pepsin treatment greatly enhanced subsequent staining of leptospires in naturally infected bovine and porcine tissues as well as in artificially infected tissues. Leptospires in naturally infected bovine tissues were usually undetectable in untreated sections but clearly visible in stained pepsin-treated sections. Naturally infected porcine kidney usually contained high levels of leptospiral antigen which could be stained without prior pepsin treatment. However, pepsin treatment of porcine tissues greatly increased the amount of leptospiral antigen detectable and made individual leptospires more conspicuous. The staining method could employ a single antiserum for the staining of leptospires from 13 serogroups. Also, leptospires could be stained in tissues stored in formalin for more than 14 months and in 26-year-old paraffin embedded tissues.

A serological survey of leptospirosis in Gippsland dairy farmers.

Serological testing for leptospirosis was performed on sera from 1074 persons who were attending the 1983 Gippsland Field Days. The most common serological reactions were against representative serovars from serogroups Hebdomadis (12%), Panama (2%), Tarassovi (2%) and Pomona (1%). The reactions against serogroup Hebdomadis were attributed to exposure to serovar hardjo; the milking of dairy cows was found to be a major risk factor. The high prevalence of seropositive results among clinically normal dairy farmers highlights the need for caution in interpreting the results of laboratory testing for leptospirosis.

Bovine abortion associated with Leptospira interrogans serovar hardjo infection.

During 1981, 265 bovine abortions were investigated by serological and histological methods for evidence of leptospiral infection. Leptospires were demonstrated in the tissues of 10 foetuses by a Levaditi silver impregnation technique. Serological testing of maternal sera indicated that Leptospira interrogans serovar hardjo was associated with 5 of the abortions while the remaining 5 were due to L. interrogans serovar pomona infection. In cases of abortion associated with L. interrogans serovar hardjo leptospires were readily demonstrated in foetal liver, kidney, intestine and heart. They were demonstrated less often in lung and placenta and could not be found in foetal brain. Autolysis did not appear to interfere with the demonstration of leptospires by silver impregnation. No lesions attributable to leptospiral infection were seen in placentas but mild interstitial nephritis was found in some of the foetuses. Fourteen other cows had serological evidence of recent leptospiral infection but leptospires were not detected in foetal tissues. Histological examination of silver impregnated foetal tissues in combination with the microscopic agglutination test was shown to be an effective method for diagnosing abortion associated with L. interrogans serovar hardjo in cattle.