Leptospirosis: I. Clinical investigation of the infection in dairy cattle in the Waikato district of New Zealand

An investigation was made into the prevalence of leptospiral infection in cattle. An area 50 km radius was selected in a region where leptospirosis was reputedly common. Farmers volunteered 250 herds with 39 500 cows for testing and 7 500 animals were selected and sampled. Twenty-nine cows (0.4%) on 14 (5.6%) of the farms had leptospiruria at the first examination. Leptospirae were cultured from the urines of nine of these animals and all were Leptospira interrogans serovar hardjo. Serologically 12.5% of cows had titres of 1:200 or greater to hardjo and 3.5% titres of 1:200 or greater to pomona. In the Spring of 1977, there was evidence of clinical leptospirosis in calves associated with only one of the herds and no clinical leptospirosis in the 250 lactating herds, although leptospiral titres were found in 88% of them. This indicated that clinical disease was much less common than infection. We concluded that leptospirosis was of minor economic importance in dairy cattle, although it could be significant in individual herds, and a health hazard to farm workers.     

Leptospirosis: II. Investigation of clinical disease in dairy cattle in the Waikato district of New Zealand

Investigations were carried out in 1975, 1976 and 1977 in 16 dairy herds where leptospiral abortions were suspected and in five other herds where clinical disease was not present. Both Leptospira interrogans serovars pomona and hardjo were isolated from cattle in herds with leptospirosis, but only pomona was recovered from those that had aborted. There was no evidence that hardjo caused clinical disease in dairy cattle in the Waikato district.

It was found that 73% of the cows that aborted and 19% of other animals in the same herds had microscopic agglutination test titres to pomona of 1:2 000 or greater. By contrast, only 2% of cattle in herds without clinical evidence of leptospirosis had such titres. One cow retained a titre of 1:2 000 or greater to pomona for 7 months; titres of this order had a shorter duration in other cows. Leptospiruria occurred in 50% of cows that had aborted and in 9% of in-contact cows in the same herds. Only 0.7% of cows had leptospiruria in the herds with no clinical disease. Ten of 35 cows shedding pomona still had leptospiruria one month later.

It was concluded that clinical leptospirosis should be diagnosed by testing a sample of the herd, rather than just individual cows, because of the variability and persistence of leptospiruria and serological titres in cows with and without clinical signs. Although hardjo is common in cattle in the Waikato district, it was not found to cause abortion in cattle.     

Epidemiology of leptospirosis in dairy farm workers in the Manawatu. Part II. A case-control study of high and low risk farms

Subsequent to a cross-sectional serological survey of Manawatu dairy farm workers, a case-control study was carried out to investigate the correlation between titres to leptospiral serovars in workers and those in cattle in their herds. A total of 52 herds was investigated, 25 of which were ‘high risk’ where milkers had titres of 1:96 or greater, and 27 were case-controls where milkers had no detectable agglutinin titres at a minimum serum dilution of 1:24.

The serological prevalence of titres to hardjo in cattle on ‘high risk’ farms (76.5%) was significantly higher (P<0.05) than on the case-control farms (60.0%). The geometric mean titres of seropositive cattle on ‘high risk’ farms were also significantly higher (P<0.0l) than in the cattle from the case-control farms, especially in the younger cohorts. These findings suggest that there was active endemic hardjo infection in the two- to three-year-old cattle on the ‘high risk’ farms.

Titres to pomona were demonstrated in only 5.2% of the cattle from both types of farm. Workers with titres to pomona tended to be from farms on which stock, especially calves, were bought-in and pigs were kept.

Conventional measures for protecting milkers from contact with infected urine appeared to be ineffective and it is concluded that prevention of leptospirosis in dairy farm workers can only be achieved by elimination of infection in the herd by vaccination of cattle.     

Leptospirosis: II. Investigation of clinical disease in dairy cattle in the Waikato district of New Zealand

Investigations were carried out in 1975, 1976 and 1977 in 16 dairy herds where leptospiral abortions were suspected and in five other herds where clinical disease was not present. Both Leptospira interrogans serovars pomona and hardjo were isolated from cattle in herds with leptospirosis, but only pomona was recovered from those that had aborted. There was no evidence that hardjo caused clinical disease in dairy cattle in the Waikato district. It was found that 73% of the cows that aborted and 19% of other animals in the same herds had microscopic agglutination test titres to pomona of 1:2,000 or greater. By contrast, only 2% of cattle in herds without clinical evidence of leptospirosis had such titres. One cow retained a titre of 1:2,000 or greater to pomona for 7 months; titres of this order had a shorter duration in other cows. Leptospiruria occurred in 50% of cows that had aborted and in 9% of in-contact cows in the same herds. Only 0.7% of cows had leptospiruria in the herds with no clinical disease. Ten of 35 cows shedding pomona still had leptospiruria one month later. It was concluded that clinical leptospirosis should be diagnosed by testing a sample of the herd, rather than just individual cows, because of the variability and persistence of leptospiruria and serological titres in cows with and without clinical signs. Although hardjo is common in cattle in the Waikato district, it was not found to cause abortion in cattle.     

Leptospira interrogans serovar hardjo Infection in Cattle in the South Okanagan District of British Columbia

An outbreak of leptospirosis due to Leptospira interrogans serovar hardjo in the South Okanagan District of British Columbia was investigated. The infection was associated primarily with bulls, but serovar hardjo was isolated from both bulls and cows at slaughter. Kidney and cerebrospinal fluid were found to contain leptospires, independently of the presence and level of serum agglutinins. Treatment of a bull twice in six months with dihydrostreptomycin failed to diminish an agglutinin titer (1/200) which persisted for two years without reexposure of the bull. A serological survey of cull cows sold through a central auction mart revealed the presence of hardjo agglutinins in 15.4% of 1300 sera representing 163 herds in 20 locations. Thirty percent of these herds contained reactor cattle. The number of premises from which reactor cattle came in a given locality varied from 4% to 67.7%. Measures to control leptospirosis in the study are suggested.     

A review of laboratory techniques and their use in the diagnosis of Leptospira interrogans serovar hardjo infection in cattle

SUMMARY This paper reviews the laboratory diagnosis of Leptospira hardjo infection in cattle. Two genotypes of L hardjo, Hardjoprajitno and Hardjobovis, have been identified in cattle, but only Hardjobovis has been isolated in Australia. There are problems with diagnosis and control of bovine leptospirosis. Infection is usually subclinical and the serological titres vary greatly in peak and duration. Leptospires may be excreted in urine for up to 18 months. Low microscopic agglutination test titres may be significant in unvaccinated herds as indicators of endemic infection. Vaccines differ in their composition, and their efficacy is difficult to evaluate. The serological response after vaccination is difficult to differentiate from the response after infection. Pregnant cows that become infected may abort, but this is usually after the serological response has peaked. Therefore, paired serum samples are of little use in diagnosing abortion caused by L hardjo. Fluorescent antibody techniques are more sensitive than dark field microscopy for detection of leptospires in urine and tissue samples. Techniques for culture have improved but are still difficult to perform and take 3 months or longer for results to be known. DNA probes and polymerase chain reaction tests are very sensitive and specific, quick to perform, and can be used on fluid and tissue samples.     

Hairy shaker disease of lambs

A serological survey of 213 randomly chosen dairy farm residents in the Manawatu showed that 66 (34%) of the people who milked cows had leptospiral titres ≤ 1:24 by the Microscopic Agglutination Test. Forty-eight (72.7%) of these people had titres to hardjo, while 29 (43.9%) had titres to pomona. Dual hardjo/pomona titres occurred in 12 people. Ballum and Copenhageni accounted for 8% of the titres found. Women milkers and farm residents who did not milk were all serologically negative. A third of the seropositive milkers had a history of clinical leptospirosis. Other factors which significantly correlated with leptospiral titres included the time spent in the dairy shed during milking, the wearing of shorts, the keeping of pigs for sale, and the number of years the individual had been working on a dairy farm. The type of milking shed and the size of the herd were interrelated and both showed strong trends towards a correlation with serological prevalence.     

SEROLOGICAL TITRES OF DAIRY COWS OVER A 63-WEEK PERIOD FOLLOWING NATURAL INFECTION WITH LEPTOSPIRA INTERROGANS SEROVAR HARDJO

SUMMARY Leptospirosis due to Leptospira interrogans serovar hardjo, occurred in a dairy herd of 104 cows and produced signs of mastitis, characterised by a sudden decrease in milk production and uniformly flaccid udders, in 11 cows. The microscopic agglutination (MA) test was used to determine antibody titres to serovar hardjo in all cows in the herd at 6, 33 and 63 weeks after the initial outbreak. The prevalence of MA antibodies to hardjo at week 6 was highest in cows in the youngest age groups and lowest in cows in the oldest age group. Over the 63-week period MA antibody titres to hardjo declined in 54 out of 62 seropositive cows, remained unchanged in 6 cows, and increased slightly in 2 cows. Fourteen of 59 cows (23.7%) with MA titres greater than 100 at week 6 had titres greater than 100 by week 63. Included in this group of 14 were 5 of 7 cows that had been affected with atypical mastitis. The distribution of serological titres to hardjo showed bimodal configurations at weeks 6 and 33 with the second peaks occurring at titres greater than 100, while the configuration was linear at week 63. MA titres to hardjo were also determined for 22 heifers 1, 16 and 36 weeks after they were added to the herd. No clinical signs of leptospirosis were observed in the heifers; however, the distribution of MA titres showed bimodal configurations at weeks 16 and 33 with second peaks occurring at titres greater than 100. Leptospiras were isolated from the urine of 2 seropositive heifers 16 weeks after their introduction to the herd, and cross-agglutinin absorption tests performed on one of the isolates indicated that it was identical to serovar hardjo.     

The epidemiological interpretation of serological responses to leptospiral serovars in sheep.

Serum samples were examined for evidence of leptospiral agglutinins from 928 sheep from 45 lines and kidneys from 12 of these lines for evidence of leptospiral infection. All sheep had been submitted for slaughter at meat works in the Manawatu.

Serological results were analysed using the results at a minimum serum dilution in the microscopic agglutination test (MAT) of 1:24 and at a minimum dilution of 1:48. It was shown that a minimum dilution of 1:24 resulted in many non-specific or cross-reactions. A minimum dilution of 1:48 was more accurate for detecting the serological prevalence of specific agglutinins to leptospires in ovine sera. Twenty percent of the sheep had titres of 1:48 or greater to hardjo, 3.8% to pomona, 2.6% to tarassovi, 2.3% to copenhageni and 2.7% to ballum. No titres of 1:48 or greater to australis were detected. Serovar hardjo was isolated from the kidneys of three animals in one line.

Eighteen months later 291 serum samples and 95 urine samples were collected from live animals on the property from which the three hardjo infected animals originated. No titres to hardjo were detected in the sera of lambs, but a serological prevalence of 44% and 84% to this serovar was demonstrated in the hoggets and ewes respectively. No leptospires were demonstrated in any of the urine samples.

These results show that sporadic infection of sheep with hardjo can occur but they also indicate that infection with this serovar is not endemic and that sheep are unlikely to act as maintenance hosts for hardjo in New Zealand.     

Phage typing of staphylococci isolated from dairy cows in New Zealand

Four red deer calves (Cervus elaphus) died with severe nephritis apparently associated with infection by Leptospira interrogans serovar pomona. The sera of 12 in-contact red deer calves were examined for leptospiral agglutinins and nine showed titres to pomona consistent with recent infection. Two also showed titres of 1:100 to serovar hardjo. The urine of five of these in-contact calves was examined periodically over a period of nine months. All were initially leptospiruric, four being infected with pomona and one with hardjo. In four animals leptospiruria could only be detected for up to six months, but one animal infected with pomona was leptospiruric for at least eight months.

The apparent source of infection was from infected cattle, and it is suggested that deer are unlikely to act as maintenance hosts for serovar pomona.     

Haemolytic disease associated with Leptospira Interrogans serovar pomona in red deer calves (Cervus elaphus)

Three red deer calves (Cervus elaphus) died with a haemolytic disease associated with infection by Leptospira interrogans serovar pomona.

Infection within the herd was more prevalent than disease. Sera from 16 herd mates were tested by the microscopic agglutination test (MAT) and 12 had leptospiral titres, the majority to serovar pomona. A few calves had titres to balcanica and hardjo.

Urine was obtained for culture from six of these calves and serovar pomona was isolated from five with titres to pomona, and hardjo from one with a litre to hardjo but not pomona.

A fourth calf died with severe nephritis but a diagnosis of leptospirosis was not confirmed in this case.     

Experimental infection of the possum (Trichosurus vulpecula) with Leptospira interrogans serovar balcanica I. Characteristics of infection

The serological, bacteriological and histopathological characteristics of experimental infection with serovar balcanica in possums (Trichosurus vulpecula) are described, and the possum is shown to be a potential maintenance host for this organism. Serum agglutination titres were maintained at almost constant levels for longer than a year, and leptospiruria was present in 50% of animals for a similar period. A paradoxical reaction to hardjo antigen was found in sera from all possums infected with balcanica.

Comparative studies were conducted using recently isolated field strains of serovars hardjo and ballum. Young possums seronegative to Hebdomadis group titres were insusceptible to challenge with hardjo, and the pathogenesis of ballum infection was characteristic of leptospiral infection in an accidental host.

The occurrence of antibodies in the urine of possums infected with balcanica is also described.     

Hypomagnesaemia in cattle

Leptospirosis is an important animal and human health problem in New Zealand, and the epidemiological and ecological aspects of infection in domestic species and free-living species are described. The characteristics of infection with a particular serovar are often considerably different in different species of animal and primary epidemiological investigations involve differentiating between maintenance-host populations and accidental-host populations for the serovars present in a particular ecosystem.

Cattle and pigs are maintenance hosts for hardjo and pomona respectively. Balcanica is maintained by possums (Trichosurus vulpecula) and ballum is maintained by the house mouse (Mus musculus), ship rat (Rattus rattus) and hedgehog (Erinaceus europaeus). In some regions, foci of copenhageni infection are maintained by Norway rats (Rattus norvegicus). Each serovar shows a high degree of parasitic adaption to its maintenance-host species and this is reflected in the nidality of leptospirosis in intensive farming environments. Accidental infections in domestic animals and man are the consequence of ecological associations with maintenance-host populations for particular serovars.

Control of leptospirosis in domestic animals depends on decreasing the prevalence of infection with serovars maintained within populations of domestic animals, and decreasing the degree of ecological association with free-living maintenance-host populations. Vaccination of cattle against hardjo. will considerably reduce the prevalence of infection in vaccinated herds and therefore decrease the prevalence of accidental hardjo infections in man.     

A cross-sectional pilot study to estimate the prevalence of and risk factors for leptospirosis in South-Western Victorian dairy herds, 2017

Leptospirosis is a zoonosis, found worldwide, affecting many species of animals. We conducted a cross-sectional study to estimate the prevalence of Leptospira borgpetersenii sv Hardjo and Leptospira interrogans sv Pomona in cattle in dairy herds in South-Western Victoria, Australia. Fifty-three herds were enrolled in the study. Urine samples were collected from 15 late-lactation cows in each herd. A questionnaire was provided to herd managers at the time of each herd visit, asking them to describe the methods they used for controlling leptospirosis, including vaccination. Urine samples were pooled at the herd level and tested for leptospira spp. using real time PCR. Urine samples from individual cows within the positive pooled samples were then tested for Leptospira Hardjo and Leptospira Pomona using qPCR. Four of the 53 herds showed positive leptospirosis results giving an apparent prevalence of 8 (95% CI 2–18) leptospira-positive herds per 100 herds at risk. Based on the 53 completed questionnaires, leptospirosis vaccination programs were not compliant with label directions in 36 of the 52 vaccinated herds: 69 (95% CI 55–81) of 100 herd managers that routinely vaccinated for leptospirosis did not comply with label directions. One herd was completely unvaccinated. Based on our findings, we estimate that approximately 10% of dairy farms in South-Western Victoria are likely to be infected with leptospirosis. While most herds are vaccinating for leptospirosis, most are not doing so according to label directions. We conclude that herd managers need to be better educated regarding leptospirosis vaccination programs.     

Transmission and control of leptospiral infection in a beef cattle herd in southern alberta

Leptospiral antibodies were detected in unvaccinated cattle on a 17 000 hectare ranch in the arid southeast region of Alberta. Antibody to serovar hardjo was present before the breeding season in 7% of 42 yearling bulls, 86% of 29 two year old bulls and 5% of 519 cows. Pomona antibody was confined to 3.7% of the cows. Bulls were treated once with dihydrostreptomycin, 25 mg/kg. Bulls and cows were vaccinated twice at a six week interval, with pomona-hardjo-gripptotyphosa bacterin before breeding and cows were revaccinated the next year. Leptospires were demonstrated in urine, kidney and spinal fluid of vaccinated and treated cattle. New infections occurred on range in vaccinates. Eighteen months after the last vaccination, hardjo and pomona antibody prevalences in cows were 3.6 and 3.2% respectively. A group of 250 seronegative cows on the same ranch were not vaccinated. They remained seronegative throughout the 2.5 years of the study. These cows, in contrast to infected groups, were excluded from pastures adjacent to perimeter herds and grazing leases, and they were bred by artificial inseminstion. Rotation through pastures in common with infected groups, and exposure to seropositive heat detector bulls, did not result in seroconversion in these cows. The study showed the potential of range bulls to amplify and transmit hardjo infection, limitations to the value of treatment and vaccination with available agents, and the potential of management practices to maintain an uninfected herd in close proximity to cattle carrying hardjo infection.     

An unusual serological response in cattle after use of a commercial leptospiral combined hardjo-pomona vaccine

Vaccination of four calves with Leptavoid (Wellcome New Zealand Limited) gave rise to Leptospira interrogans serovars hardjo and pomona microscopic agglutination test titres that could not be distinguished in magnitude from post-infection titres. Vaccination of four calves with Lepto-3 (ICI Tasman Limited) gave rise to much lower titres.

Revaccination of cows with Leptavoid caused a rise in hardjo titres which was significantly greater than after use of Lepto-3.

The possibility that titres were due to the simultaneous infection with serovars pomona and hardjo of only the animals vaccinated with Leptavoid must he discounted.     

Strategies of the control of an outbreak of leptospiral infection in dairy cattle in Northeastern Brazil

The aim of the present study was to describe the strategies of the control of an outbreak of leptospiral infection in dairy cattle in Maranhão State, Northeastern Brazil. In the period from January to July 2015, 18 (17%) out of 106 cows presented abortion, six (5.7%) stillbirth, and 12 (11.3%) repeated estrus, totaling 24 animals with reproductive problems. The diagnosis of leptospirosis was based on serology (microscopic agglutination test—MAT), bacteriological culture, and polymerase chain reaction (PCR). Antibiotic therapy, vaccination protocols, and changes in management practices were suggested as control measures. Of all animals on the farm (n?=?280), 136 (48.6%) were seropositive for at least one serovar of Leptospira sp. No pure leptospiral culture was obtained. Eight of the animals with reproductive problems yielded positive PCR results (vaginal fluid of seven animals and urine and vaginal fluid of one animal). Genetic sequencing of a vaginal fluid/urine PCR-positive sample revealed Leptospira borgpetersenii. One year after the adoption of control measures, no reproductive problems were observed. Thus, leptospirosis probably caused the reproductive failures in the herd, and the control and prevention measures implemented were efficient in controlling the disease.

     

Seroprevalence of and Risk Factors for Leptospiral Antibodies among Cattle in the State of Yucatan,Mexico

Sera obtained from cattle in the state of Yucatan, Mexico, were screened using the microscopic agglutination test against 13 serovars of Leptospira interrogans. A total of 62.8% (461/734) cows were positive for one or more serovars. This seroprevalence probably reflects infection because vaccination against leptospirosis has not been practised in Yucatan. The most common antibodies detected were those against antigens of serovars hardjo (54.1%) and tarassovi (53.3%). Region was the only risk factor associated with the seroprevalence of leptospirosis (p < 0.05). This revised version was published online in July 2006 with corrections to the Cover Date.     

Epidemiology of leptospirosis in dairy farm workers in the Manawatu. Part II. A case-control study of high and low risk farms

Subsequent to a cross-sectional serological survey of Manawatu dairy farm workers, a case-control study was carried out to investigate the correlation between titres to leptospiral serovars in workers and those in cattle in their herds. A total of 52 herds was investigated, 25 of which were high risk where milkers had titres of 1:96 or greater, and 27 were case-controls where milkers had no detectable agglutinin titres at a minimum serum dilution of 1:24. The serological prevalence of titres to hardjo in cattle on high risk farms (76.5%) was significantly higher (P<0.05) than on the case-control farms (60.0%). The geometric mean titres of seropositive cattle on high risk farms were also significantly higher (P<0.01) than in the cattle from the case-control farms, especially in the younger cohorts. These findings suggest that there was active endemic hardjo infection in the two- to three-year-old cattle on the high risk farms. Titres to pomona were demonstrated in only 5.2% of the cattle from both types of farm. Workers with titres to pomona tended to be from farms on which stock, especially calves, were bought-in and pigs were kept. Conventional measures for protecting milkers from contact with infected urine appeared to be ineffective and it is concluded that prevention of leptospirosis in dairy farm workers can only be achieved by elimination of infection in the herd by vaccination of cattle.     

Evaluation of a recombinant LigB protein of Leptospira interrogans serovar Canicola in an enzyme-linked immunosorbent assay for the serodiagnosis of bovine leptospirosis

A recombinant leptospiral lipoprotein, LigB, was evaluated for use in the diagnosis of bovine leptospirosis by enzyme-linked immunosorbent assay (rLigB IgG ELISA). The standard reference test (Microscopic agglutination test, MAT) of 200 serum samples from cattle suspected of leptospirosis showed that 95 (47.5%) samples had positive agglutination titres, which ranged from 100 to 1600. In rLigB IgG ELISA, 49% of the samples were positive. Sensitivity of IgG ELISA for 95 bovine sera, which had MAT titres of greater than or equal to 100, were 100%. ELISA showed a specificity of 97.1% with 105 bovine sera, which were negative at a 1:50 dilution in MAT for Leptospira interrogans serovars. The results of ELISA and MAT correspond very good. When analytical specificity of IgG ELISA was evaluated using bovine serum samples from animals showing the serum antibodies to other pathogens, no cross-reaction was observed. Thus the recombinant LigB IgG ELISA can be used instead of the MAT as an aid to the diagnosis of bovine leptospirosis.