Vaccination practices for Leptospira spp. on New Zealand dairy farms.

AIMS: To describe leptospiral vaccination practices in dairy herds in New Zealand and evaluate conformity with best practice guidelines issued by the New Zealand Veterinary Association using data from a questionnaire administered by participating veterinary practices. METHODS: A cross-sectional study of 200 randomly selected dairy farms stratified by herd size and region throughout New Zealand was conducted from January to April 2016 to investigate leptospiral vaccination practices in dairy herds in New Zealand. Using a pre-tested questionnaire administered during a face-to-face interview, vaccination practice details such as vaccine types, time, and age of vaccination and whether vaccines were administered by veterinary or farm staff, were collected. RESULTS: Leptospiral vaccination programmes had been implemented on 199/200 (99.5 (95% CI = 97.2-99.9)%) farms, and on 178 (89.4%) of those, programmes had been running for ≥5 years. Most farmers used bivalent vaccines containing antigens for leptospiral serovars Pomona and Hardjo (144/179 (80.4%) in calves, 112/167 (60.7%) in heifers, and 112/163 (68.7%) in cows), rather than trivalent vaccines which also include antigens for L. interrogans serovar Copenhageni. In total, 123/200 (61.5%) of farmers purchased only vaccinated animals but 51/199 (25.6%) were unsure of the vaccination status of purchased cattle. Sixty-one percent (105/172) of farmers had other livestock on their farms and of them, 78/186 (42%) vaccinated some or all for Leptospira spp. Leptospiral vaccines were administered always or sometimes with other animal remedies on 30/190 (15.8%) and 91/190 (47.9%) of farms, respectively. Most farmers had not made changes to their vaccination programme in the previous 5 years. Timing of first vaccination of calves ranged from 2 weeks to 10 months of age, with 112/189 (59.3%) vaccinating by 4 months of age. Approximately half of the farms followed the best practice guideline for the timing of vaccinations for calves (high-risk farms; 67/162; 41.4%) heifers (72/165, 43.6%), and cows (171/184; 92.9%). CONCLUSIONS: The results of this survey suggest that there is almost universal adoption of leptospiral vaccination for dairy cattle in New Zealand. However, there remain areas for improvement regarding the proportion of farmers following best practice guidelines and refinement of vaccination programmes, particularly with respect to timing of vaccination in calves.

Serological study of Leptospira interrogans serovar Copenhageni and L. borgpetersenii serovars Tarassovi and Ballum in beef cattle, sheep and deer in New Zealand.

AIMS: To estimate animal-level seroprevalence of Leptospira interrogans serovar Copenhageni and L. borgpetersenii serovars Ballum and Tarassovi, in beef cattle, sheep and deer on New Zealand farms, and herd/flock-level seroprevalence of any serovar when existing same-sera data for serovars Hardjobovis and Pomona were included, and to determine associations between risk factors and animal-level seroprevalence. METHODS: Banked sera from sheep (n = 82), beef (n = 54) and deer (n = 62) herds/flocks (n = 3,878 animals) from seven regions were analysed using the microscopic agglutination test. Titres of ≥48 were designated positive. Herds/flocks were considered positive if either ≥1, ≥2 or ≥3 animals were positive. Existing same-sera data for serovars Hardjobovis and Pomona were included to establish farm-level any-serovar seropositivity. Factors associated with serological status were analysed using generalised estimating equations. RESULTS: Animal-level seroprevalence for serovars Ballum, Copenhageni, and Tarassovi, respectively, was 13.7 (95% CI = 11.7-16.0)%, 12.6 (95% CI = 10.6-14.7)% and 18.0 (95% CI = 15.7-20.5)% for beef cattle, 10.5 (95% CI = 9.0-12.1)%, 16.7 (95% CI = 14.9-18.6)% and 14.0 (95% CI = 12.4-15.8)% for sheep and 6.6 (95% CI = 5.3-8.2)%, 15.5 (95% CI = 13.5-17.7)% and 3.6 (95% CI = 2.7-4.8)% for deer, respectively. Herd/flock-level seroprevalence for Ballum was 86.6, 52.4 and 39.0% for sheep, 85.2, 52.7 and 33.3% for beef cattle and 50.8, 27.9 and 21.3% for deer at definitions ≥1, ≥2 and ≥3 seropositive animals per species, respectively. For Copenhageni, corresponding data were 95.1, 73.2 and 56.1% for sheep, 68.5, 48.2 and 29.6% for beef cattle and 73.8, 57.4 and 41.0% for deer, and for Tarassovi, 80.5, 59.7 and 45.1% for sheep, 83.3, 68.5 and 61.1% for beef cattle, and 42.6, 16.4 and 4.9% for deer. Seropositivity to all serovars was observed from all regions, with some differences in seroprevalence observed between species and regions, but not between islands. Combining with Hardjobovis and Pomona data, herd/flock-level seropositivity for all animal species and all five Leptospira serovars was 100% at definition ≥1 animal positive, and 97.5 and 96.3% for sheep flocks, 87.8 and 97.8% for beef cattle herds, and 89.3 and 75% for deer herds at ≥2 and ≥3 animals positive, respectively. CONCLUSIONS: Seropositivity to serovars Ballum, Copenhageni and Tarassovi is common in sheep, beef cattle and deer New Zealand and most, or all farms have ≥1 livestock species seropositive to ≥1 serovar. CLINICAL RELEVANCE: Serovars Ballum, Tarassovi and Copenhageni should be considered when clinical or subclinical signs of leptospirosis are observed in sheep, beef cattle or deer. Livestock sector workers are potentially at risk of exposure.

On-farm risk factors associated with Leptospira shedding in New Zealand dairy cattle.

This study aimed to evaluate risk factors associated with shedding of pathogenic Leptospira species in urine at animal and herd levels. In total, 200 dairy farms were randomly selected from the DairyNZ database. Urine samples were taken from 20 lactating, clinically normal cows in each herd between January and April 2016 and tested by real-time polymerase chain reaction (PCR) using gyrB as the target gene. Overall, 26.5% of 200 farms had at least one PCR positive cow and 2.4% of 4000 cows were shedding Leptospira in the urine. Using a questionnaire, information about risk factors at cow and farm level was collected via face-to-face interviews with farm owners and managers. Animals on all but one farm had been vaccinated against Hardjo and Pomona and cows on 54 of 200 (27%) farms had also been vaccinated against Copenhageni in at least one age group (calves, heifers and cows). Associations found to be statistically significant in univariate analysis (at P < 0.2) were assessed by multivariable logistic regression. Factors associated with shedding included cattle age (Odds ratio (OR) 0.82, 95% CI 0.71-0.95), keeping sheep (OR 5.57, 95% confidence interval (CI) 1.46-21.25) or dogs (OR 1.45, 95% CI 1.07-1.97) and managing milking cows in a single as opposed to multiple groups (OR 0.45, 95% CI 0.20-0.99). We conclude that younger cattle were more likely to be shedding Leptospira than older cattle and that the presence of sheep and dogs was associated with an increased risk of shedding in cows. Larger herds were at higher risk of having Leptospira shedders. However, none of the environmental risk factors that were assessed (e.g. access to standing water, drinking-water source), or wildlife abundance on-farm, or pasture were associated with shedding, possibly due to low statistical power, given the low overall shedding rate.

Emerging Leptospira strain poses public health risk for dairy farmers in New Zealand.

Leptospira infection in dairy cattle and leptospirosis in dairy farm workers were common in New Zealand prior to the introduction of dairy cattle vaccination in the 1980s. Despite widespread vaccination within the dairy industry, the long-term effectiveness of vaccination and current Leptospira exposure status remained unknown. A cross-sectional study was conducted from January-April 2016 to investigate the prevalence of pathogenic Leptospira spp. DNA in urine at cow and herd level, and its relationship to five Leptospira serovars known to be endemic. Two hundred dairy farms were randomly selected from the national database. Twenty paired blood and urine samples were collected on each farm from adult cows (n = 4000). Sera were tested using the Microscopic Agglutination Test against serovars Hardjobovis (termed Hardjo), Pomona, Copenhageni, Ballum and Tarassovi with titres ≥48 being considered positive. Urine was tested using quantitative real-time PCR (qPCR) that amplifies the gryB gene. All but one herd had been vaccinated with a bivalent Hardjo/Pomona or trivalent vaccine incorporating Copenhageni. In total, 2.4% of cows were urine qPCR positive and 27% of farms had at least one urine qPCR positive cow. Overall 63% of cows were seropositive to one or more serovars: 44% for Hardjo, 28% for Pomona, 15% for Copenhageni (in vaccinated herds), and for unvaccinated cows: 1% for Copenhageni, and 3% for Ballum and 17% for Tarassovi. Of the 94 qPCR urine-positive cows, 51 were seropositive to Tarassovi, 3 to Ballum, 3 to Copenhageni, 24 to Hardjo, and 17 to Pomona, the latter two presumably reflecting vaccination titres. A strong association was found between shedding and serology for Tarassovi. While there was no evidence that current vaccination programmes were ineffective in protecting against their target serovars, serovar Tarassovi has apparently emerged in NZ dairy cattle. As Tarassovi is currently not included in vaccines and is prevalent in notified leptospirosis cases in dairy workers, we concluded that this serovar poses a public health risk.

Seroprevalence and herd-level risk factors for seroprevalence of Leptospira spp. in sheep, beef cattle and deer in New Zealand.

AIMS To determine seroprevalence of Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona in beef cattle, sheep and deer in New Zealand and the association between farm-level risk factors and seroprevalence. METHODS Between June 2009 and July 2010, 20 serum samples per flock or herd were collected from 162 sheep flocks, and 116 beef cattle and 99 deer herds from 238 farms, along with farm data by interview. Samples were tested for antibodies to serovars Hardjo and Pomona by microscopic agglutination testing, with a titre ≥48 being positive. Species-specific associations between herd-level seroprevalence (number of seropositive animals, for each serovar, divided by the number of animals tested) and herd-level risk factors were determined by multivariable logistic regression analysis. Vaccinated animals were excluded from seroprevalence estimates but included in multivariable analyses. RESULTS For sheep (n=3,339), animal-level seroprevalence was 43.6 (95% CI=41.9-45.3)% for serovar Hardjo and 14.1 (95% CI=12.9-15.3)% for serovar Pomona; for beef cattle(n=1,886), it was 45.6 (95% CI=43.3-47.9)% for Hardjo and 19.6 (95% CI=17.9-21.5)% for Pomona; and for deer (n=1,870), it was 26.3 (95% CI=24.3-28.4)% for Hardjo, 8.8 (95% CI=7.6-10.2)% for Pomona. In sheep flocks (n=161), flock-level prevalence for Hardjo varied from 77.9-91.3%, and for Pomona from 40.4-73.9%, when ≥1, ≥2 or ≥3 animals were seropositive. In beef herds (n=95), herd-level prevalence for Hardjo varied from 79.0-90.5%, and for Pomona from 42.1-68.4%. In deer herds (n=93), herd-level prevalence for Hardjo varied from 45.2-59.1%, and for Pomona from 22.6-48.4%. For sheep flocks, herd-level seroprevalence for Hardjo was associated with flock size (OR=1.56) and number of dogs (OR=0.75), and for Pomona, seroprevalence varied with region. For beef cattle, herd-level seroprevalence for Hardjo was associated with herd size (OR=1.4), presence of dams (OR=0.6) and vaccination (OR=2.9), and for Pomona, co-grazing with deer (OR=0.4), vaccination (OR=3.22), presence of dams (OR=0.2) and streams (OR=2.7). For deer herds, seroprevalence for Hardjo or Pomona was associated with herd size (OR=1.6 and 1.8) and varied with region, and for Pomona seroprevalence varied with season (summer vs. winter: OR=4.8). CONCLUSIONS Serovars Hardjo and Pomona were highly prevalent at herd and animal levels, with serovar Hardjo highest in all species. Larger herd size was the common risk factor for seroprevalence in all livestock species.

Meta-analysis of the efficacy of Leptospira serovar Hardjo vaccines to prevent urinary shedding in cattle.

Leptospirosis is a zoonosis often associated with occupational exposure from livestock that can be prevented by animal vaccination. Several trials have assessed vaccine efficacy in livestock but there have been no attempts to evaluate these trials jointly. This systematic review and meta-analysis aimed to estimate vaccine efficacy to prevent urinary shedding of Leptospira serovar Hardjo (Hardjo) in cattle. Three databases were used to search for relevant papers published from 1980 to 2015 evaluating commercial vaccines to prevent urinary shedding of leptospires after artificial conjunctival or natural challenge. A total of 1237 articles were initially identified. Eight articles containing information from nine trials that assessed vaccine efficacy to prevent Hardjo urinary shedding, as per bacteriological culture, were included in the meta-analysis. Fixed effects Mantel-Haenszel (MH) and a Bayesian random effects meta-analyses were used to estimate the efficacy of vaccination to prevent Hardjo shedding in urine. Vaccine efficacy against Hardjo challenge was 88.7% (95% CI 81.0%-93.2%) in the MH meta-analysis and 89.9% (95% probability interval 80.6%-94.9%) in the Bayesian random effects meta-analysis. There was no evidence of heterogeneity of study results (p = 0.17). The estimated vaccine efficacy to prevent urinary shedding of Hardjo in cattle may be sufficient to reduce disease incidence in animals and exposure risk of people working in close contact with cattle.

Data on Leptospira interrogans sv Pomona infection in Meat Workers in New Zealand.

The data presented in this article are related to the research article entitled "Comparison between Generalized Linear Modelling and Additive Bayesian Network; Identification of Factors associated with the Incidence of Antibodies against Leptospira interrogans sv Pomona in Meat Workers in New Zealand" (Pittavino et al., 2017) [5]. A prospective cohort study was conducted in four sheep slaughtering abattoirs in New Zealand (NZ) (Dreyfus et al., 2015) [1]. Sera were collected twice a year from 384 meat workers and tested by Microscopic Agglutination for Leptospira interrogans sv Pomona (Pomona) infection, one of the most common Leptospira serovars in humans in NZ. This article provides an extended analysis of the data, illustrating the different steps of a multivariable (i.e. generalized linear model) and especially a multivariate tool based on additive Bayesian networks (ABN) modelling.

Comparison between generalized linear modelling and additive Bayesian network; identification of factors associated with the incidence of antibodies against Leptospira interrogans sv Pomona in meat workers in New Zealand.

BACKGROUND: Additive Bayesian Network (ABN) is a graphical model which extends Generalized Linear Modelling (GLM) to multiple dependent variables. The present study compares results from GLM with those from ABN analysis used to identify factors associated with Leptospira interrogans sv Pomona (Pomona) infection by exploring the advantages and disadvantages of these two methodologies, to corroborate inferences informing health and safety measures at abattoirs in New Zealand (NZ). METHODOLOGY AND FINDINGS: In a cohort study in four sheep slaughtering abattoirs in NZ, sera were collected twice a year from 384 meat workers and tested by Microscopic Agglutination with a 91% sensitivity and 94% specificity for Pomona. The study primarily addressed the effect of work position, personal protective equipment (PPE) and non-work related exposures such as hunting on a new infection with Pomona. Significantly associated with Pomona were "Work position" and two "Abattoirs" (GLM), and "Work position" (ABN). The odds of Pomona infection (OR, [95% CI]) was highest at stunning and hide removal (ABN 41.0, [6.9-1044.2]; GLM 57.0, [6.9-473.3]), followed by removal of intestines, bladder, and kidneys (ABN 30.7, [4.9-788.4]; GLM 33.8, [4.2-271.1]). Wearing a facemask, glasses or gloves (PPE) did not result as a protective factor in GLM or ABN. CONCLUSIONS/SIGNIFICANCE: The odds of Pomona infection was highest at stunning and hide removal. PPE did not show any indication of being protective in GLM or ABN. In ABN all relationships between variables are modelled; hence it has an advantage over GLM due to its capacity to capture the natural complexity of data more effectively.

Seroprevalence and Risk Factors for Leptospira Seropositivity in Beef Cattle, Sheep and Deer Farmers in New Zealand.

Leptospirosis is a global zoonosis that in New Zealand affects primarily people occupationally exposed to livestock. The objective of this study was to estimate the seroprevalence of five Leptospira serovars in farmers working on cattle, sheep and deer farms that had the serological status of animals previously assessed and to identify risk factors for farmer seropositivity. A total of 178 farmers from 127 properties participated in the study. Blood samples were tested using the microscopic agglutination test (MAT) for the presence of antibodies to Leptospira. Samples with a MAT titre ≥48 were considered seropositive. Using Bayesian statistical analysis, the median seroprevalence of Leptospira, all serovars combined, was estimated to be 6.6% (95% probability interval (PI) 3.6-10.9%). Risk factors associated with seropositivity were assisting deer or cattle calving, farming deer, having ≥25% of flat terrain and high abundance of wild deer on farm, while high possum abundance on farm was negatively associated with seropositivity. No association was observed between farmer serostatus and previously recorded livestock serology. Leptospira seropositivity was associated with influenza-like illness of farmers (RR = 1.7; 95% PI 1.0-2.5). Assuming a causal relationship, this suggested an annual risk of 1.3% (95% PI 0.0-3.0%) of influenza-like illnesses due to Leptospira infection in the population of farmers. The association between seropositivity and disease can be used to estimate the public health burden of leptospirosis in New Zealand. Identifying and understanding risk factors for Leptospira seropositivity can inform preventive measures, hence contributing to the reduction of leptospirosis incidence in farmers.

Serological patterns, antibody half-life and shedding in urine of Leptospira spp. in naturally exposed sheep.

AIMS: To determine within-farm prevalence, longitudinal pattern of exposure measured by serology, antibody titre longevity and point prevalence of shedding in urine of Leptospira borgpetersenii serovar Hardjo and L. interrogans serovar Pomona in naturally infected sheep on a sample of commercial farms in New Zealand. METHODS: On eight commercial sheep farms, between September 2011 and January 2014, blood samples were collected from 115-217 ewe lambs on each farm, at intervals of 2-11 months. They were analysed by microscopic agglutination test (MAT) for antibodies to L. borgpetersenii serovar Hardjo and L. interrogans serovar Pomona, using a titre cut-point of 48. Urine from 98 animals was tested by quantitative PCR (qPCR). The half-life of antibodies was estimated in 185 sheep for serovar Hardjo and 21 for Pomona, and the seroprevalence and mean titre of animals lost to follow-up was compared with those remaining in the study. RESULTS: Within-flock seroprevalence for serovar Hardjo reached a maximum at 17-22 months of age, ranging from 79 to 100%. Seroprevalence for serovar Pomona rose above 10% on three farms and increased to 21-54% by 4-14 months. Seroconversions occurred mainly from late autumn to early summer at 7-15 months of age. Seroprevalences ranging from 3 to 76% for serovar Hardjo and 0.5 to 15% for serovar Pomona were observed up to 3 months of age, likely due to maternally derived immunity. The half-life of antibody in response to infection was estimated to be 6.7 (95% CI=5.8-7.9) months for serovar Hardjo and 6.3 (95% CI=4.8-9.0) months for Pomona. The prevalence of sheep with urine positive for leptospires on qPCR on each farm ranged from 11 to 88%. All but one of the qPCR-positive animals were seropositive for serovar Hardjo. On two farms where Pomona exposure was observed, animals that were lost to follow-up had a higher geometric mean titre for serovar Pomona than those remaining in the study. CONCLUSIONS: This study demonstrated seasonal exposure from autumn to early summer in young sheep, a wide range of within-flock serological and shedding prevalence, and gives an estimation of the half-life of MAT titres in sheep. More extensive data are needed to fully understand the epidemiology of leptospirosis in sheep flocks across New Zealand and, along with economic analysis, to justify and design cost-effective and efficient control measures to protect human and animal health.

Prevalence and risk factors for Leptospira exposure in New Zealand veterinarians.

This study assessed seroprevalence and risk factors for Leptospira (serovars Hardjo, Pomona, Ballum, Copenhageni, Tarassovi) exposure in New Zealand veterinarians. Veterinarians (n = 277) at one of two conferences were voluntarily enrolled and blood samples taken. Microscopic agglutination test (MAT) titres ⩾48 were considered seropositive. Fourteen veterinarians (5·1%, 95% confidence interval 2·8-8·3) were seropositive to Leptospira. Home slaughter of cattle or pigs were significant risk factors for Leptospira exposure. There were no clear relationships between the animal species handled at work and serostatus. However, veterinarians spending a 'mid to high' proportion of their time (>50% to ⩽75%) with pets had higher odds of being seropositive than those not working with pets. A borderline positive association (P = 0·09) was observed between seropositivity and clinical influenza-like illness (⩾3 days off work) in the 18 months before the study. Assuming causality, this suggests that 8·3% of these cases may be attributed to Leptospira exposure.

Risk of infection and associated influenza-like disease among abattoir workers due to two Leptospira species.

The aims of this study were to determine the annual incidence of infection with Leptospira interrogans serovar Pomona and/or Leptospira borgpetersenii serovar Hardjo and its association with influenza-like illness (ILI) in meat workers in New Zealand. Sera were collected twice, 50-61 weeks apart, from 592 workers at eight abattoirs slaughtering sheep (n = 4), cattle (n = 2) and deer (n = 2), and tested by the microscopic agglutination test for Hardjo and Pomona. Forty-nine (8·3%) participants either seroconverted or had at least a twofold increased serological titre against either serovar. The worker infection risk was higher in sheep abattoirs (11·9%) than in abattoirs processing deer (0%) or cattle (1·2%) (P < 0·01). The annualized risk of mild (ILI) or severe clinical disease attributable to the two Leptospira serovars was 2·7%. This study has demonstrated that meat workers are at substantial risk of infection and clinical disease, suggesting further investigation of infection sources and preventive measures are warranted.

Risk factors for new infection with Leptospira in meat workers in New Zealand.

OBJECTIVE: To evaluate risk factors for new infection with Leptospira interrogans sv Pomona and Leptospira borgpetersenii sv Hardjo in meat workers. METHODS: Sera were collected twice approximately 12 months apart from 592 workers from eight abattoirs slaughtering sheep, cattle or deer and tested by microscopic agglutination for Pomona and Hardjo. Information on potential risk factors were recorded and analysed by multivariable logistic regression. RESULTS: Forty-nine (8.3%) participants, either seroconverted or had at least a titre increase by two dilutions against either serovar. While in sheep meat workers, the annual infection risk was 11.3% (95% CI 8.5% to 14.8%), in deer meat workers it was 0% (95% CI 0.0% to 10.9%) and in those processing beef cattle, 1.2% (95% CI 0.2% to 4.6%). Risk factors for new infection in sheep abattoirs were worker position, abattoir and time worked in the meat industry. The new infection risk was highest at the beginning of the slaughter line (stunning and hide removal; relative risk, RR 7.5, 95% CI 2.5 to 22.4), followed by positions on the line involving the removal of high-risk material (bladder, and kidneys; RR 5.2, 95% CI 1.7 to 16.0). Risk was lower in the offal/pet food area (RR 4.1, 95% CI 1.0 to 16.4), and lowest in the boning room or office. Wearing personal protective equipment did not reduce the risk of new infection. CONCLUSIONS: This study has demonstrated ongoing exposure to leptospires in meat workers and risk factors for challenge. We recommend measures such as improvement of personal protective equipment use, changes in slaughter procedure or vaccination of sheep against Leptospira to reduce the risk.

Shedding and seroprevalence of pathogenic Leptospira spp. in sheep and cattle at a New Zealand Abattoir.

A cross-sectional study was carried out on sheep and cattle slaughtered at a New Zealand abattoir from September to November 2010 to investigate the supplier-specific shedding rate, renal carriage rate and seroprevalence of leptospires. In the 2008/2009 season, this abattoir experienced three human leptospirosis cases from 20 staff, of which two were hospitalized. Urine, kidney and blood samples were collected from carcasses of 399 sheep (six suppliers, 17 slaughter lines) and 146 cattle (three suppliers, 22 slaughter lines). The urine and kidney samples were tested by quantitative real-time PCR (qPCR), while serum samples (from coagulated blood samples) were tested by microscopic agglutination test (MAT). In total, 27% (73/274; 95% CI: 18-37) of urine samples tested positive by qPCR. Species-specific shedding rates (prevalence of positive urine qPCR) were 31% (95% CI: 17-48) for sheep and 21% (95% CI: 14-30) for cattle. For 545 kidney samples tested, 145 were qPCR positive (27%; 95% CI: 17-39). The average prevalence of kidney qPCR positivity was 29% (95% CI: 17-45) for sheep and 21% (95% CI: 15-28) for cattle. Three hundred and thirty of 542 sampled sheep and cattle had antibodies against Leptospira borgpetersenii serovar Hardjobovis (Hardjobovis) and/or Leptospira interrogans serovar Pomona (Pomona), based on reciprocal MAT titre ≥1 : 48 (overall seroprevalence of 61%; 95% CI: 48-73). Seroprevalence was 57% (95% CI: 40-72) for sheep and 73% (95% CI: 59-83) for cattle. Among the seropositive animals, 41% (70/170; 95% CI: 30-54) were shedding (tested positive by urine qPCR) and 42% (137/330; 95% CI: 30-54) had renal carriage (tested positive by kidney qPCR). Some risk management options for abattoirs or farms to prevent human leptospirosis infections include vaccination of maintenance hosts, the use of personal protective equipment, and the application of urine qPCR to detect shedding status of stock as surveillance and as an alert.

A serological survey of leptospiral antibodies in dogs in New Zealand.

AIM: To investigate the prevalence of titres to four endemic leptospiral serovars in dog sera from the lower half of the North Island, and the South Island of New Zealand submitted to diagnostic laboratories, and to explore the association between the prevalence of seropositive samples to leptospirosis and breed group, age group and sex. METHODS: Serum samples from 655 dogs residing in the central and lower North Island and from the South Island of New Zealand were sourced from the Massey University Veterinary Teaching Hospital and from submissions to New Zealand Veterinary Pathology in 2005. They were screened by the Microscopic Agglutination Test (MAT) against Leptospira interrogans serovars Copenhageni and Pomona and L. borgpetersenii serovars Hardjo and Ballum. Titres greater or equal to 96 were considered positive. Variables investigated for their association with the prevalence of seropositive samples to leptospirosis included serovar, breed, North vs. South Island, age and sex. RESULTS: Positive MAT titres to Leptospira interrogans serovar Copenhageni were found in 10.3 % of dogs (95% CI=8.1-12.9), and were more common than positive titres to other leptospiral serovars. Small breeds did not have a lower prevalence of Copenhageni titres than other breeds. Positive titres to Leptospira borgpetersenii serovar Hardjo were associated with breeds of dogs used as farm working dogs. There was no significant difference in the prevalence of positive leptospiral titres between dogs from the North or South Islands. Dogs greater than 12 years of age were less likely to have positive titres to Leptospira than younger dogs. No association was found between positive titres and sex. CONCLUSIONS: Breeds of dogs used as farm working were at greater risk of exposure to Leptospira borgpetersenii serovar Hardjo. Small breeds did not have a lower risk of seropositivity to Copenhageni than farm working breeds. Further study should be undertaken to confirm the prevalence of positive titres to leptospirosis in farm dogs and dogs resident in the South Island. CLINICAL RELEVANCE: The risk of dogs being exposed to Leptospira interrogans serovar Copenhageni, and requirement for vaccination against serovar Copenhageni, cannot be determined by geographical location or breed group. Vaccination against Leptospira borgpetersenii serovar Hardjo is likely to be beneficial in working dogs.

Growth response and shedding of Leptospira spp. in urine following vaccination for leptospirosis in young farmed deer.

AIM: To investigate the effect of vaccination against Leptospira serovars Hardjo-bovis and Pomona on growth rate and shedding of leptospires in urine in rising 1-year-old farmed red deer. METHODS: In early March 2007, 230 female and 205 male, 3-month-old deer on five farms were treated with streptomycin then were randomly allocated to a control group (n = 218), or were vaccinated (n = 217) with a bivalent whole-cell killed leptospiral vaccine (Leptavoid-2) followed by a booster 4 weeks later. These animals were isolated from other 3-month-old deer on each property until May, when all vaccinated and control deer were combined with deer not treated with streptomycin, for maximum exposure to natural leptospiral challenge. Seroconversion was monitored in control deer against Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona using the microscopic agglutination test (MAT). A cut-off of ≥ 1:48 was used to identify a seropositive response to infection, and ≥1:24 to indicate a response to vaccination. Urine was collected from females in all groups and tested for shedding of leptospires using bacterial culture and real-time PCR. Male deer from vaccinated and control groups were weighed every 2-3 months to measure growth rate. RESULTS: In vaccinated female deer, antibody was present to Hardjo-bovis in 39-73% and to Pomona in 78-100% of animals in May, with titres ranging between 1:24-1:96 and 1:24-1:1536, respectively; control female deer were all serologically negative. In November, female control deer were seropositive to Hardjo-bovis on 4/5 farms (seroprevalence range 4-78%) but seronegative to Pomona on all farms. Between March and November mean growth rate was higher in vaccinated than control deer (26 g/day) on one farm (p = 0.049) that had the highest prevalence of seroconversion (78-88%) and urinary shedding (56%). Between August and November on 4/5 farms, when almost all seroconversions occurred, growth rates were higher in vaccinated than control deer (31 g/day; p = 0.011). Urinary shedding of leptospires was detected by culture and/or PCR in 8/34 (24%) unvaccinated control and 20/38 (53%) animals not treated with streptomycin on 2/5 farms; no shedding was detected in vaccinated deer. The average risk difference for shedding between vaccinated and control deer was -11% (95%CI = -18 to -4) (p = 0.001). CONCLUSION: Vaccination against Leptospira serovars Hardjo-bovis and Pomona was associated with higher liveweight gain and reduced urinary shedding of leptospires in young deer naturally challenged by Hardjo-bovis. The association with liveweight gain was evident on some but not all farms.

Utilizing a combination of molecular and spatial tools to assess the effect of a public health intervention.

Until recently New Zealand had one of the highest rates of human campylobacteriosis reported by industrialized countries. Since the introduction of a range of control measures in the poultry production chain a reduction in human cases of around 50% has been observed nationwide. To inform risk managers a combination of spatial, temporal and molecular tools - including minimum spanning trees, risk surfaces, rarefaction analysis and dynamic source attribution modelling - was used in this study to formally evaluate the reduction in disease risk that occurred after the implementation of control measures in the poultry industry. Utilizing data from a sentinel surveillance site in the Manawatu region of New Zealand, our analyses demonstrated a reduction in disease risk attributable to a reduction in the number of poultry-associated campylobacteriosis cases. Before the implementation of interventions poultry-associated cases were more prevalent in urban than rural areas, whereas for ruminant-associated cases the reverse was evident. In addition to the overall reduction in prevalence, this study also showed a stronger intervention effect in urban areas where poultry sources were more dominant. Overall a combination of molecular and spatial tools has provided evidence that the interventions aimed at reducing Campylobacter contamination of poultry were successful in reducing poultry-associated disease and this will inform the development of future control strategies.

Vaccination for leptospirosis improved the weaning percentage of 2-year-old farmed red deer hinds in New Zealand.

AIM: To investigate the effect of leptospiral vaccination against serovars Hardjo-bovis and Pomona on fetal loss and weaning percentage in rising 2-year-old farmed red deer hinds. METHODS: In mid-February 2007, 252 rising 2-year-old hinds on four farms received a single dose of streptomycin (25 mg/kg), to minimise leptospiral infection. They were randomly allocated to vaccinated and control groups. Vaccinated hinds (n=125) received a 2-ml S/C injection of a bivalent whole-cell killed leptospiral vaccine (Leptavoid-2) followed by a booster 4-6 weeks later, and were grazed with control hinds (n=127). These animals were isolated from other hinds on each property, until after mating (June 2007), when all vaccinated and control hinds were combined with hinds not treated with streptomycin, for maximum exposure to natural leptospiral challenge. Evidence of natural challenge by Leptospira spp. was assessed in blood samples from control hinds by serology against L. borgpetersenii serovar Hardjo-bovis and L. interrogans serovar Pomona, using the microscopic agglutination test (MAT), and in hinds not treated with streptomycin by detection of shedding of organisms in urine, using bacterial culture and real-time PCR. Pregnancy diagnosis was carried out in May/June 2007, using transrectal ultrasonography, to determine conception. In late October, prior to calving, the pregnant vaccinated and control hinds were examined by palpation of the abdomen and udder, to determine the percentage of hinds pregnant at term and assess fetal loss. In March 2008, at weaning, vaccinated and control hinds were examined for lactation status, using observation and palpation of the udder. The differences between the groups were evaluated using matched logistic regression analysis. RESULTS: After mating, pregnancy was diagnosed in 97/125 (77.6%) vaccinated and 106/127 (83.5%) control hinds. All four farms had serological evidence of Hardjo-bovis infection, and a single hind was serologically positive for Pomona between October and March. Real-time PCR confirmed urinary shedding on two farms. The mean percentage of hinds pregnant at term, for those animals confirmed pregnant after mating, in the vaccinated and control groups was 95/97 (98%) (range 95-100%) and 103/106 (97%) (range 94-100%), respectively (p>0.05). The mean weaning percentage for vaccinated and control groups was 86/97 (89%) (range 78-95%) and 88/106 (83%) (range 76-88%), respectively (p=0.015). CONCLUSION: Vaccination for leptospirosis resulted in no difference in the percentage of hinds pregnant at term, but a higher weaning percentage compared with unvaccinated controls suggesting that vaccination reduced perinatal and/or pre-weaning mortality.

Serosurvey of leptospirosis and investigation of a possible novel serovar Arborea in farmed deer in New Zealand.

AIM: To investigate the prevalence of Leptospira spp. and possible novel serovar Arborea infection in farmed deer in New Zealand. METHODS: In September 2006, five serum samples from a serum bank from each of 70 farms sampled for a previous national prevalence survey were forwarded to the World Health Organisation/Food and Agriculture Organisation/World Organisation for Animal Health (WHO/FAO/OIE) reference laboratory for leptospirosis in Brisbane, Australia, to test for reactivity to a reference panel of 23 serovars, most believed to be exotic to New Zealand, using the microscopic agglutination test (MAT). Eleven farms were seropositive for Arborea, a serovar novel to New Zealand. In July 2007, 126 additional banked serum samples from nine of those 11 farms (n=8-20/farm) were sent to the reference laboratory for similar serology. Two farms in the Southland region were considered positive for serovar Arborea. Tissue from deer kidneys (n=43) from these two farms collected at a deer slaughter premises (DSP) was cultured in November 2007 and November 2008. Sera from those deer were also sent to the laboratory in Brisbane. RESULTS: From the initial 350 sera, 96 (27.4%) and 19 (5.4%) samples were positive for Leptospira borgpetersenii serovar Hardjo-bovis and Leptospira interrogans serovar Pomona respectively. There were cross-reactions between serovar Hardjo-bovis with serovars Medanensis and Szwajizak. Serological evidence of serovars Tarassovi, Grippotyphosa, Celledoni, Australis, Zanoni, Robinsoni, Canicola, Kremastos, Bulgarica, Cynopteri, Ballum, Bataviae, Djasiman, Javanica, Panama, Shermani and Topaz was negative or sporadic, generally with titres of 1:50 and therefore likely non-specific. Fourteen (4.0%) samples from 11 farms were positive for serovar Arborea, justifying further investigation. The prevalence of serovar Arborea was 15% and 30% on two farms, from the 126 samples. None of 43 kidney and serum samples collected subsequently from those two farms were positive by culture or serology for serovar Arborea. CONCLUSIONS: While there were samples serologically positive for serovar Arborea in deer, attempts to isolate the organism were unsuccessful. The sample size for the follow-up investigation was insufficient to validate the presence or absence of infection, so further study should be undertaken to verify the status of this serovar of Leptospira spp. in New Zealand, in both deer and other livestock species.

Assessment of occupational exposure to leptospirosis in a sheep-only abattoir.

This study estimated the frequency of exposure of meat workers to carcasses infected with Leptospira serovars Hardjobovis or Pomona in a sheep-only abattoir in New Zealand. A stochastic spreadsheet model was developed to assess the daily risk of exposure of eviscerators, meat inspectors and offal handlers to live leptospires in sheep carcasses from May to November 2004 (high-risk period), and from December 2004 to June 2005 (low-risk period). The average sheep processed per day were 225 for an eviscerator, 374 for a meat inspector, and 1123 for an offal handler. The median daily exposures during high- and low-risk periods were 11 [95% distribution interval (DI) 5-19] and three (95% DI 1-8) infected carcasses/day for eviscerators, 18 (95% DI 9-29) and six (95% DI 2-12) for meat inspectors, and 54 (95% DI 32-83) and 18 (95% DI 8-31) for offal handlers, respectively. Stochastic risk modelling provided evidence that processing of sheep carcasses exposed meat workers regularly to live leptospires with substantial seasonal variation.