Leptospirosis associated with serovars hardjo and pomona in red deer calves (Cervus elaphus)

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.     

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.     

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.     

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.     

Prevention of Leptospira interogans serovar pomona infection in cattle

A commercial hardjo-pomona vaccine which has previously been shown to be effective against hardjo infection was tested against pomona. Following challenge all 11 six-month-old non-vaccinated calves seroconverted and pomona was isolated from blood or urine on at least one occasion from nine of them. Pomona was isolated once only, on the third day after challenge, from the blood of one of 11 vaccinated calves.     

EVALUATION OF A HARDJO-POMONA VACCINE TO PREVENT LEPTOSPIRURIA IN CATTLE EXPOSED TO A NATURAL CHALLENGE WITH LEPTOSPIRA INTERROGANS SEROVAR HARDJO

SUMMARY The efficacy of a vaccine against Leptospira interrogans serovar pomona and Leptospira interrogans serovar hardjo was evaluated in a group of dairy heifers that were serologically negative at the time of vaccination and later subjected to natural challenge with L. interogans serovar hardjo. Thirty-nine heifers were vaccinated twice, at a one-month interval, with a commercially prepared bivalent vaccine, while 43 unvaccinated heifers were used as controls. After vaccination, microscopic agglutination (MA) titres of serums to L. interrogans serovar hardjo ranged from 32 to 512, and those to L. interrogans serovar pomona ranged from 32 to 2048. Titres resulting from vaccination were short-lived and after the first vaccination the serums of 95% of vaccinated heifers did not react in the MA test by 24 weeks. The first indication of infection in the heifers was noted at week 6, and by week 16, elevated MA titres (≥128) to L. interrogans serovar hardjo had occurred in 62% of unvaccinated heifers and had increased to 85% by week 24. At week 18, 18% of the vaccinated heifers and 56% of the unvaccinated heifers had leptospiruria (p<0.01); after 22 weeks, 13% of the vaccinated heifers and 58% of the unvaccinated heifers showed evidence of leptospiruria (p<0.01).     

Protection of cattle against natural challenge with Leptospira interrogans serovar hardjo using a hardjo-pomona vaccine

The hardjo component of a bivalent hardjo/pomona vaccine prepared from cultures grown in a protein-free medium was tested in calves which experienced a natural challenge about 7 months after vaccination. A significant degree of protection was obtained using either seroconversion or positive urine cultures as in-dicators of infection. Seroconversion occurred in 10/10 non-vaccinates and 2/9 vaccinates (p <0.01), Leptospires of serovar hardjo were isolated from the urine of 6/10 non-vaccinated calves and 0/9 vaccinated animals (p <0.05).     

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.     

Serological reactions against Leptospira interrogans serovars in New Zealand horses

Extract

In October 1989, we received a blood sample from a horse which had recently aborted. The sample showed a very high titre against Leptospira interrogans serovar copenhageni and lower titres against several other tested serovars. Two in-contact horses showed high titres against serovar hardjo.     

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.     

A serological survey of leptospirosis in cats

A serological survey of 225 cats from different parts of the NorthIsland was conducted using 11 leptospiral serovar antigens. Twenty (8.8%) of the sera gave titres to the microagglutination test.The prevalence of titres to the serovars were as follows: five (2.22%) to copenhageni, five (2.22%) to hardjo, four (1.77%) to ballum, four (1.77%) to pomona, one (0.44%) to balanica and one (0.44%) to canicola; no titres to tarassovi, bataviae, pyrogenes and australis were recorded. It is hypothesised that prey-predator transmission between cats and rodents is responsible for infections with serovars copenhageni and ballum. Infections with the other serovars are presumed to result from contact with infected animals or a contaminated environment.     

Leptospirosa pomona Abortion Storm in a Cattle Herd in Saskatchewan

Abortions occurred in 18% of 131 beef cows and heifers during two months, on a farm in southern Saskatchewan. The losses began two weeks after acute febrile illness and agalactia in a dairy cow to which the beef herd had been exposed. A diagnosis of Leptospira interrogans serovar pomona infection was made on the basis of serology in cows and the finding of leptospires in fetal tissues by fluorescent antibody test. Tentative diagnosis of infectious bovine rhinotracheitis delayed treatment and prophylaxis until infection attained high intensity in the herd and severe losses to the farmer occurred. Abortions ceased after vaccination against pomona and oxytetracycline treatment of pregnant cows, although chronic debility followed the acute phase of the disease in some cows. Recrudescence of infection was suspected four months later, when acute agalactia occurred in one cow and debility in calves and cows was recurring. Pomona infection was not proven, but dihydrostreptomycin treatment and revaccination were applied to the whole herd. Seroconversion and IgM antibody continued to indicate a persistent source of infection and susceptibility in a minority of the population one year after onset. The source of the original infection is believed to have been a carrier beef cow, or a dairy cow which was leptospiruric at the time of contact with the beef herd. With the exception of one aborted calf, no evidence of pomona infection was found outside the farm, in cattle or wild mammals tested serologically within a radius of 30 km, during one year following the outbreak.     

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.     

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.     

The prevalence of antibodies to serovars of Leptospira interrogans in horses

SUMMARY Serum samples from 272 horses, some III, were tested by the microscopic agglutination test for the presence of antibody to 12 serovars of Leptospira interrogans. Serums from 41.5% of horses reacted to one or more of the serovars tested; the most common reactions were to L. interrogans serovar ballum (15.1%), L. interrogans serovar autumnalis (11.8%), L. interrogans serovar icterohaemorrhagiae (9.9%), L. interrogans serovar pomona (8.1%) and L. interrogans serovar hardjo (7.7%).     

Protection of pregnant swine by vaccination against leptospira infection

SUMMARY The protection conferred on pregnant gilts by 2 commercially available Leptospira interrogans serovars pomona and tarassovi bacterins was evaluated. Gilts vaccinated either 3, 6 or 12 months prior to natural challenge with L. interrogans serovar pomona had significantly lower abortion rates (2% vs 69%) and foetal mortality rates (14% vs 57%) than unvaccinated controls. One vaccine was significantly superior to the other and contained approximately twice the number of L. interrogans serovar pomona organisms per vaccine dose. Neither vaccine protected against renal colonisation but vaccination reduced urinary excretion of leptospires. Both vaccines reduced agglutinating antibody response to infection, as measured by the microscopic agglutination (MA) test. This may prevent the detection of a carrier animal by serology. Foetal pigs did not develop specific MA titres. Cultural methods were not reliable in making a diagnosis of foetal infection. Histopathology of foetal liver and kidneys helped in making a diagnosis of foetal infection.     

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 balcunica 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 titre to hardjo but not pomona. A fourth calf died with severe nephritis but a diagnosis of leptospirosis was not confirmed in this case.     

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.