Seroprevalence and association with abortion of leptospirosis in cattle in Ontario.

Sera were collected using a systematic random sampling from 348 cattle herds in Ontario, in proportion to the cattle population in different areas. One cow in five from 296 dairy herds and one in three from 52 beef herds were sampled. The sera were analyzed for prevalence of antibodies to Leptospira interrogans serovar grippotyphosa, hardjo, icterohaemorhagiae and pomona using the microscopic agglutination test. Herd seroprevalence (one or more animals with titer greater than or equal to 80) in beef and dairy herds combined was grippotyphosa 2%, hardjo 13.8%, icterohaemorrhagiae 10.1% and pomona 25.8%; 39% of all herds showed evidence of leptospiral infection with one or more serovars; 44.2% of 52 beef herds had serological evidence of infection with serovar hardjo compared to 8.4% of 296 dairy herds (P less than 0.0001). Seroprevalence of other serovars was not significantly different between beef and dairy herds. The proportion of beef animals seropositive for hardjo and for pomona increased with age, particularly for hardjo; 26.5% of beef animals aged nine years or over were seropositive for hardjo. Dairy animals showed a significant rise of hardjo but not pomona titers with age. The seroprevalence of pomona infection was significantly higher in dairy cattle in eastern Ontario than in other regions. Thirty-four (6.1%) of 553 aborted bovine fetuses had leptospires detected by immunofluorescence techniques. Sixty-five percent of these fetuses were from submissions made between November and January. Leptospires were identified as serovar hardjo by specific immunofluorescence. There appeared, however, to be a paradoxical serological response in that eight aborting cows had antibody titers to pomona rather than hardjo.(ABSTRACT TRUNCATED AT 250 WORDS)     

Diagnostic specificity, sensitivity and cross-reactivity of an enzyme-linked immunosorbent assay for the detection of antibody against Leptospira interrogans serovars pomona, sejroe and hardjo in cattle.

Outer sheath antigen was prepared from Leptospira interrogans serovars pomona, sejroe and hardjo by treating the organisms with 1.0M NaC1 followed by 0.04% sodium dodecyl sulfate (SDS). Sodium dodecyl sulfate was removed from the SDS-protein complexes by the extraction of dodecyl sulfate anions as ion pairs with triethylammonium cations into an organic solvent. The outer sheath antigen was recovered from the organic solvent as a precipitate and used as the source of leptospiral enzyme-linked immunosorbent assay (ELISA) antigen. Utilizing this antigen, ELISA was adapted to detect bovine serum antibody to L. interrogans serovars pomona, sejroe and hardjo. The specificity of this assay in 344 bovine sera, which were negative in the microscopic agglutination test (MAT) for seven serovars, was 99.4%. In sera from 37 and 87 cattle which revealed MAT titers greater than or equal to 1:50 for L. interrogans serovars pomona and sejroe, the relative sensitivity of the test was 100%. The ELISA also showed a considerable degree of low level cross-reactivity with other serovars. Sixty-six (75.9%) out of 87 bovine sera which were MAT-positive (MAT titer of greater than or equal to 1:50) with serovars sejroe and hardjo only were ELISA positive with heterologous pomona antigen; 16 (43.2%) and six 16.2%) out of 37 bovine sera which were MAT positive MAT titer of greater than or equal to 1:50) with serovar pomona only were ELISA positive with heterologous sejroe and hardjo ELISA antigen respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Survey of leptospiral agglutinins in the sera of swine of southeastern Alabama

The occurrence of leptospirosis in swine of southeastern Alabama was determined. A total of 627 sera were tested, using the microscopic agglutination method, with live antigens of 12 serovars. Of the sera tested, 121 (19.3%) had a titer of 1:100 or greater to the serovars employed. The percentage distribution of sera with titers of greater than or equal to 1:100 among serovars most commonly reported was as follows: Leptospira interrogans serovars pomona, 3.8%; icterohaemorrhagiae, 3.3%; canicola, 1.6%; hardjo, 0.7%; and grippotyphosa, 0.16%. Of the less commonly recognized leptospiral serovars, the percentages reacting were as follows: ballum, 4.9%; autumnalis, 3.2%; pyrogenes, 1.1%; and bataviae, 0.4%. None of the sera reacted with antigen of serovars australis, tarassovi, or wolffi.     

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.     

Leptospirosis in dogs: a serologic survey and case series 1996 to 2001

All leptospirosis microscopic agglutination test titers for the Leptospira serovars icterohaemorrhagiae, canicola, grippotyphosa, bratislava, hardjo, and pomona conducted on 1,260 blood samples from dogs at the University of Illinois Veterinary Diagnostic Laboratory between March 1996 and March 2001 were evaluated. Low titers (1:100 to 1:400) were predominantly L. icterohaemorrhagiae and L. canicola, which represented the predominant serovars (65.4%) among all positive samples with low titers. L. grippotyphosa was the predominant serovar (72.1%) among samples with clinically significant titers (greater than 1:800). The medical records of 87 dogs with a titer greater than 1:800 that were patients at the Veterinary Teaching Hospital of the University of Illinois were reviewed. A clinical diagnosis of leptospirosis was made in 15 cases (17.2%) based on the elevated titer, appropriate clinical signs, lack of recent vaccination, and lack of concurrent disease that could explain the clinical signs present. Renal disease was present in 10 of the cases, concurrent renal and hepatic disease in two, and hepatic disease in three. In 12 cases, the predominant serovar was L. grippotyphosa; titers to L. grippotyphosa and L. bratislava were equal in magnitude in three cases.     

Leptospirosis in dogs: a serologic survey and case series 1996 to 2001

All leptospirosis microscopic agglutination test titers for the Leptospira serovars icterohaemorrhagiae, canicola, grippotyphosa, bratislava, hardjo, and pomona conducted on 1,260 blood samples from dogs at the University of Illinois Veterinary Diagnostic Laboratory between March 1996 and March 2001 were evaluated. Low titers (1:100 to 1:400) were predominantly L. icterohaemorrhagiae and L. canicola, which represented the predominant serovars (65.4%) among all positive samples with low titers. L. grippotyphosa was the predominant serovar (72.1%) among samples with clinically significant titers (greater than 1:800). The medical records of 87 dogs with a titer greater than 1:800 that were patients at the Veterinary Teaching Hospital of the University of Illinois were reviewed. A clinical diagnosis of leptospirosis was made in 15 cases (17.2%) based on the elevated titer, appropriate clinical signs, lack of recent vaccination, and lack of concurrent disease that could explain the clinical signs present. Renal disease was present in 10 of the cases, concurrent renal and hepatic disease in two, and hepatic disease in three. In 12 cases, the predominant serovar was L. grippotyphosa; titers to L. grippotyphosa and L. bratislava were equal in magnitude in three cases.     

The significance of leptospiral titres associated with bovine abortion

To investigate relationships between serological titres to 2 serovars, pomona (L. pomona) and hardjo (L. hardjo), of Leptospira interrogans and abortions, log linear and logit models were fitted to herd and individual cow data from cattle serologically negative for brucellosis. Serological titres to both serovars were significantly related to abortions in individual cows, with L. pomona having a stronger relationship than L. hardjo. L. hardjo was not significant when herd data were analysed. Differences between dairy and beef cattle in the serological titres found to both L. pomona and L. hardjo were detected when data sets of all cattle or cattle with no history of abortion were analysed. The beef/dairy differences may be due to different management practices and/or to different geographical distributions of both serovars and populations of beef and dairy cattle. If there are no cattle in a herd with a reciprocal titre of 3000 or greater for L. pomona, it is unlikely that L. pomona is associated with the abortion problem. There was no specific L. hardjo titre which separated high and low probabilities that the serum came from a cow or herd with an abortion history.     

Prevalence of antibodies of Leptospira serovars in beef cattle in central Queensland.

OBJECTIVE: To obtain up-to-date data on the prevalence of antibodies to Leptospira serovars in central Queensland beef herds preliminary to assessing their role in bovine subfertility and the role of cattle as a zoonotic reservoir. DESIGN: Sera from 2857 female cattle in 68 central Queensland beef herds were tested for antibodies to 14 Leptospira serovars using the microscopic agglutination test. Vaccination use and age of cattle were collected to enable the calculation of crude and age-stratified seroprevalences. RESULTS: The most commonly detected antibodies were to serovars hardjo (15.8% crude seroprevalence), tarassovi (13.9%), pomona (4.0%) and szwajizak (2.2%). Vaccinates were omitted from the hardjo and pomona seroprevalence data. The seroprevalence for hardjo and pomona tended to increase with age of the animals. CONCLUSION: These results are broadly similar to those of previous serological surveys. The data suggest that serovars other than hardjo, pomona and tarassovi, are unlikely to have a significant role in bovine subfertility and that cattle are unlikely to be a source of human infection with them in central Queensland.     

Serological prevalence of bovine leptospirosis in Plateau State, Nigeria

Serum samples obtained from 1.537 cattle in the 14 local government areas (LGAs) of Plateau State of Nigeria were screened for the presence of leptospiral antibodies using 13 serovars in a modified microscopic agglutination test (MAT). Two hundred and twenty-two (14.4 p.100) of the cattle tested had leptospiral antibody titres of 1:100 or higher to one or more of the test antigens. The prevalence rates of antibodies to individual serovars were: hardjo (35.6 p.100), pomona (11.7 p.100), pyrogenes (11.7 p.100), canicola (9.5 p.100), grippotyphosa (7.7 p.100), bratislava (5.9 p.100), icterohaemorrhagiae (5.9 p.100), ballum (4.5 p.100), autumnalis (3.6 p.100), bataviae (2.3 p.100) and tarassovi (1.8 p.100). The serological prevalence of bovine leptospirosis in the various local government areas of Plateau State of Nigeria differed significantly (P less than 0.05; X2).     

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.     

Serological titers to various leptospiral serovars before and after vaccinating gilts with three commercial vaccines

The antibody response to various leptospiral serovars was evaluated in six-month-old gilts vaccinated with three commercial vaccines, each containing five leptospiral serovars.

All three vaccines elicited a significant postvaccinal antibody response to Leptospira canicola, grippotyphosa and icterohaemorrhagiae (copenhageni). The antibody response to the other leptospiral antigens in the vaccines varied among the vaccinates. None of the vaccinated groups developed significant titers to L. hardjo. Two of the three vaccines elicited a significant postvaccinal response to L. pomona, but in each case the titer mean of the vaccinated group was <1/100. Although not among the antigens in the vaccines, titers to L. bratislava increased in all vaccinated groups, except one, and in one control group.

     

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.     

Experimental infection of calves with Leptospira interrogans serovar hardjo: conjunctival versus intravenous route of exposure

Eight-month-old calves, housed under maximum isolation, were exposed to pathogenic Leptospira interrogans serovar hardjo by the conjunctival route or IV. One calf served as an unexposed control. Infection was monitored serologically (microscopic agglutination test and enzyme-linked immunosorbent assay; ELISA) and by leptospiral culture isolation from periodic urine samples and from the kidneys, epididymides, and aqueous humor collected at slaughter. Microscopic agglutination test titers of greater than or equal to 1:40 were detected among all IV exposed calves at postinoculation day (PID) 7 and among conjunctival exposed calves at PID 14. By ELISA, all IV exposed calves were positive by PID 3, whereas conjunctival exposed calves were positive at PID 14. The ELISA was more sensitive for the detection of antibodies against leptospires in cattle. Leptospires were isolated from the urine of 4 calves and from the kidney of 3 calves exposed by the conjunctival route, but not from IV exposed calves. The results indicated that the conjunctival route of exposure was a more natural and successful route for experimental infection of cattle with serovar hardjo.     

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.     

Herd-level risk factors associated with Leptospira spp. seroprevalence in dairy and beef cattle in Spain.

Our aim in this cross-sectional study was to investigate the seroprevalence of Leptospira spp. infection in herds and cattle and the relationships between seroprevalence and beef versus dairy, size, replacement policy and grazing management in a representative area of beef- and dairy-cattle production in Spain. Herds were the initial sampling unit. Blood samples were collected from 762 dairy cattle belonging to 81 herds and 1238 beef cattle from 134 herds; sera were tested for antibodies against 11 serovars of Leptospira (autumnalis, ballum, bratislava, canicola, castellonis, copenhagheni, grippotyphosa, hardjo, louisiana, pomona and tarassovi) using the microagglutination test. Forty-three percent (36.2-49.5%) of the herds and 8% (6.4-8.8%) of the individuals were seropositive against one or more of the serovars studied. Bratislava was the most-prevalent serovar (24% of the herds and 4% of the individuals) followed by hardjo (11 and 1%, respectively). Grippotyphosa, copenhagheni and tarassovi were more prevalent in dairy than in beef herds (P<0.001, P<0.05, P<0.05, respectively) -- but no significant association was found between herd-size and Leptospira seroprevalence for any of the serovars considered.     

Is canine leptospirosis underdiagnosed in southern Ontario? A case report and serological survey

An eight-year-old city-dwelling Cairn Terrier was presented to a veterinary hospital in acute renal failure with evidence of hepatic insufficiency. The dog was treated symptomatically over three days, during which time vomiting was largely controlled, but it became jaundiced as hepatic insufficiency worsened. Leptospira pomona was demonstrated in large numbers by immunofluorescent staining of urinary sediment. It was isolated and its identity confirmed as L. pomona genotype kennewicki. The source of the infection was thought to be raccoons.

Sera from 474 blood samples submitted for diagnostic purposes to two clinical pathology laboratories in southern Ontario were examined with the microscopic agglutination test for antibodies to selected leptospiral serovars. Of the sera tested, 39.2% reacted at titers ≥1:100 with one or more serovars, the majority of all sera (26.2%) reacting at low titers to canicola or icterohaemorrhagiae, or both. These reactions likely resulted from vaccination. A smaller proportion reacted to other serovars tested: autumnalis (3.8%), bratislava (8.2%), grippotyphosa (1.9%), hardjo (3.0%), and pomona (3.2%). Among dogs reacting to these latter serovars (other than bratislava), many had broadly cross-reacting and relatively high titers. One dog with a titer of 1:800 to pomona had had a disease typical of leptospirosis two years previously. Three other dogs with high titers to autumnalis, bratislava, or mixed serovars had clinical histories compatible with leptospirosis.

We suggest that leptospiral bacterins for dogs in Ontario be broadened to include at least serovars autumnalis and pomona.

     

LEPTOSPIRAL AGGLUTININS IN DOGS IN SYDNEY

From June 1971 to June 1972, sera from 600 dogs in Sydney were tested for leptospiral agglutinins by a rapid slide agglutination method. The end-point titre was taken at 50 percent agglutination of the live organisms. Forty-one samples (6.8 percent) had a significant leptospiral titres (100 or greater) and 5 of these reacted to 2 serotypes. Thirty serums (5 percent) contained agglutinins against L. copenhageni, and 6 (1 percent) against L. pomona, while a few samples reacted against hardjo, tarassovi, australis, grippotyphosa or pyrogenes serotypes. No significant titres were found to L. canicola, L. hebdomadis, L. autumnalis or L. bataviae.     

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 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.     

Comparison of polymerase chain reaction assay,bacteriologic culture,and serologic testing in assessment of prevalence of urinary shedding of leptospires in dogs

OBJECTIVE: To compare results of polymerase chain reaction (PCR) testing of urine samples, serologic testing, and bacteriologic culture of urine to determine prevalence of urinary shedding of leptospires in dogs. DESIGN: Serial case study. ANIMALS: 500 dogs evaluated serially without regard to health status. PROCEDURE: Urine samples were examined via PCR assay and bacteriologic culture for leptospires. Blood samples were analyzed for antibodies against serovars canicola, bratislava, pomona, icterohemorrhagiae, grippotyphosa, and hardjo. RESULTS: Titers > or = 1:100 against at least 1 serovar were detected in 104 (20.8%) dogs, and titers > or = 1:400 were detected in 41 (8.2%) dogs. High titers were detected most commonly to serovar grippotyphosa, followed by icterohemorrhagiae, canicola, pomona, bratislava, and hardjo. High titers to > 1 serovar were detected in 14 dogs. A positive PCR assay result was obtained in 41 (8.2%) dogs, only 9 of which had a titer > or = 1:100. Leptospires were not cultured from the urine of any dog. Only 4 dogs had clinical leptospirosis. Overall disease prevalence was 0.8% for the 6-month evaluation period. Compared with PCR assay, serologic testing for predicting shedding had a sensitivity of 22%, specificity of 79%, positive predictive value of 9%, and negative predictive value of 92%. CONCLUSIONS AND CLINICAL RELEVANCE: Irrespective of health status, 8.2% of dogs were shedding pathogenic leptospires. Serologic testing was a poor predictor of urinary shedding. Clinically normal dogs that shed leptospires may pose a zoonotic risk to their owners.