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An epidemiological investigation was conducted in an unvaccinated dairy farming enterprise in which three workers on one of the milking herds (Herd 1) were diagnosed with leptospirosis due to serovars Hardjo (H) (n = 2) and Pomona (P) (n = 1) between January and March 2015. Blood and urine samples were collected from milking cows in Herd 1 (N = 230) and Herd 2 (N = 400), rising one- (R1, N = 125) and rising two-year-old (R2, N = 130) replacement heifers, and four pigs associated with Herd 1, in March 2015. Sera were tested using the MAT for serovars H, P, Copenhageni (C), Ballum (B) and Tarassovi (T), and urine samples were tested by qPCR. Seventy-five per cent of 109 cows in Herd 1 and 36% of 121 in Herd 2 were seropositive (≥48), predominantly to H and P, and 23% of 74 cows in Herd 1 and 1% of 90 cows in Herd 2 were qPCR positive. Fifty-five per cent of 42 R2 heifers were seropositive to T. No R1 and 17% of 42 R2 heifers were qPCR positive. Subsequently, all cattle were vaccinated for H and P, and Herds 1 and 2 were given amoxicillin. After the booster vaccination, 7% of 91 in Herd 1, 2% of 82 in Herd 2 and 11% of 38 R1 heifers (sampled as R2) were PCR positive. After the amoxicillin treatment, no cows in Herd 1 and 5% of 62 cows in Herd 2 were urine PCR positive. Calves and pigs were seropositive to H, P, C and B. Vaccination and antibiotic treatment appeared effective in reducing the risk of exposure of workers to vaccine serovars. However, evidence of non-vaccine serovars indicated that workers likely remain at risk of exposure to Leptospira., (© 2019 Blackwell Verlag GmbH.)

In New Zealand, up to 97% of NZ sheep flocks are seropositive to Leptospira borgpetersenii serovar Hardjo and/or Leptospira interrogans Pomona, yet vaccination is rare. This study evaluated the impact of exposure to these serovars and of vaccination on sheep growth. One third of 2260 ewe lambs on eight farms were randomly selected and vaccinated with a primary and booster bivalent Hardjo and Pomona vaccine starting at one month of age on seven farms and at around five months of age on one farm. Repeated blood samples were taken over one (n = 6 farms, bred as ewe lambs at 7-8 months of age) or two (n = 2 farms, bred as rising 2-year-old ewes) years and tested by microscopic agglutination test to assess exposure to Hardjo and Pomona. Individual weights were recorded at the same time and modelled using a multilevel linear model accounting for within-farm clustering and repeated measures. Predicted average weights were computed and compared based on the vaccination status and within the control group based on exposure status (positive for Hardjo only, Pomona only, Hardjo and Pomona and negative) for each combination of farm and weighing episode. Statistical significance of the comparison was evaluated after adjustment for multiple comparisons. There was no difference in average weight between vaccinated and control sheep before or after vaccination in any of the flocks. The comparison between sheep seropositive for either or both serovars and seronegative sheep was inconclusive, with variations of direction and magnitude of the difference between farms and weighing episodes. In the absence of an overall growth response to vaccination, widespread adoption of vaccination would unlikely yield an economic response at the industry level. However, the inconsistency observed when comparing animals based on their exposure status suggests that the actual effect of leptospirosis on growth is difficult to predict. A study of the effect on sheep reproduction is needed to fully assess the effect of vaccination on sheep production., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Most New Zealand sheep flocks are seropositive to Leptospira serovars Hardjo and/or Pomona, yet vaccination is rare. This study evaluated the impact of exposure to these serovars and of vaccination, on primiparous one- (P1) and two-year-old (P2) sheep reproduction outcomes. The study was designed as a split-flock vaccination trial, with a third of the animals vaccinated starting at one month of age. Reproduction outcomes were the proportion of bred P1 (7 months old) and as P2 (19 months old) scanned pregnant, the proportion of pregnant ewes rearing a lamb to tail docking and the proportion of docked lambs that were weaned. Odds ratios and their 95% confidence intervals were calculated to compare reproductive performance between vaccinated and control sheep, and within the control group, between seropositive and seronegative sheep. Odds ratios (OR) were also calculated to assess the relationship between vaccination and loss to follow-up. There was no difference in pregnancy and docking rates between vaccinated and control sheep, or between seropositive and seronegative sheep. P1 with a Hardjo titre ≥1536 were significantly less likely (OR = 0.41, 95%CI = 0.19-0.93) to keep a lamb between docking and weaning than P1 with both Hardjo and Pomona titres <1536, for an observed difference in weaning rate of up to 22.6% points on one farm. A reduction of weaning rates in 2-tooths seropositive for Pomona alone and both Hardjo and Pomona was observed but this was non-significant, possibly because of a lack of power. No difference in weaning rate was observed between vaccinated and control P1 or P2. On one farm vaccinated P1 were less likely to be lost to follow-up (OR = 0.27, 95% CI 0.08 to 0.95) between breeding and weaning. Comparing reproductive performance of vaccinated and control sheep revealed no significant difference. However, comparing exposed and non-exposed ewes revealed a possible adverse effect of Leptospira on weaning rates. This suggests that a full vaccination program may result in an improvement of reproductive outcomes, possibly by providing herd immunity., (Copyright © 2018 Elsevier Inc. All rights reserved.)

L. borgpetersenii serovar Hardjo and L. interrogans serovar Pomona are endemic in New Zealand sheep. An effective vaccine and vaccination strategy would protect both humans and livestock. Four to 12 lambs were selected from each of eight farms (total=84, vaccinated group), while four to 16 lambs (total=98) served as unvaccinated controls. A commercial Hardjo/Pomona vaccine was given at 1-6 weeks of age, 5-11 weeks later and 33-67 weeks later on seven farms and at 18 weeks of age and 5 weeks later on the eighth farm. Vaccinates and controls were grazed together. Blood was regularly collected from the control group to assess flock exposure. Urine was collected from both groups 26-82 weeks after the second vaccination and tested by quantitative PCR. Seroprevalence in controls at the time of urine sampling ranged from 2.7 to 98.2% for Hardjo and from 0 to 54.1% for Pomona with seroconversion occurring 13 to 67 weeks after the second vaccination in all but one farm where exposure had happened by the time of vaccination. The shedding prevalence adjusted for clustering in farms was 45.1% [95% CI 17.6-72.7] (for an observed number of 50/98) in the control animals and 1.8% [95% CI 0.0-10.1] (for an observed number of 5/84) in the vaccinated animals. The vaccine was 100% effective on five farms where animals were vaccinated before 12 weeks of age and before natural exposure occurred, but the effectiveness was 80% [0-97] on one farm where the lambs were exposed before vaccination and 65% [9-87] to 80% [0-97] on one farm where the animals were fully vaccinated by 24 weeks of age. The overall vaccine effectiveness was 86.3% [63.6-94.8%] despite maternal antibodies in some flocks at first vaccination. Vaccination timing seemed to be crucial in achieving optimum reduction in shedding in urine in vaccinated sheep., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

A SYTO9 real-time polymerase chain reaction assay for detection of pathogenic Leptospira spp. based on amplification of DNA gyrase subunit B (gyrB) gene has been optimized and evaluated for sensitivity and specificity on kidney and urine samples of New Zealand farmed deer. The detection limit was 10(3) cells/ml (2-10 copies/reaction). Comparison of the assay on deer kidneys (n = 268) with culture as the gold standard revealed a sensitivity and specificity of 85% and 99.2%, respectively. For deer urine (n = 113), the assay was compared with known inoculated samples and revealed a sensitivity and specificity of 96.7% and 100%, respectively. The assay was applied for quantifying pathogenic leptospires shed naturally in deer urine and revealed a detectable concentration of 3.7 × 10(3) to 1.7 × 10(6) cells/ml. To assess the assay's capability for identifying pathogenic Leptospira spp., 14 field isolates of L. borgpetersenii serovar Hardjo-bovis and L. interrogans serovar Pomona were amplified for polymerase chain reaction (PCR) product, purified, and sequenced. When compared with the National Center for Biotechnology Information database, sequence data matched with L. borgpetersenii serovar Hardjo-bovis in 13 samples and L. interrogans serovar Pomona in 1 sample, which was consistent with the microscopic agglutination test (MAT). Sequence analysis of purified PCR product amplified directly from kidney and urine samples also yielded serovar-comparable MAT results. Results suggest that the assay is rapid, sensitive, and specific for detection of pathogenic leptospires in deer clinical samples. The developed assay can also be used for estimating the concentration of leptospires and identifying Leptospira spp. in combination with DNA sequencing.