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    Investigating animals and environments in contact with leptospirosis patients in Aotearoa New Zealand reveals complex exposure pathways.
    (Taylor and Francis Group, 2025-02-12) Benschop J; Collins-Emerson JM; Vallee E; Prinsen G; Yeung P; Wright J; Littlejohn S; Douwes J; Fayaz A; Marshall JC; Baker MG; Quin T; Nisa S
    CASE HISTORY: Three human leptospirosis cases from a case-control study were recruited for in-contact animal and environment sampling and Leptospira testing between October 2020 and December 2021. These cases were selected because of regular exposure to livestock, pets, and/or wildlife, and sampling was carried out on their farms or lifestyle blocks (sites A-C), with veterinarians overseeing the process for livestock, and cases collecting environmental and wildlife samples. LABORATORY FINDINGS: Across the three sites, a total of 137 cattle, > 40 sheep, 28 possums, six dogs, six rats, three pigs and three rabbits were tested. Herd serology results on Site A, a dairy farm, showed infection with Tarassovi and Pomona; urinary shedding showed Leptospira borgpetersenii str. Pacifica. Animals were vaccinated against Hardjo, Pomona and Copenhageni. The farmer was diagnosed with Ballum. On Site B, a beef and sheep farm, serology showed infection with Pomona; animals were not vaccinated, and the farmer was diagnosed with Hardjo. On Site C, cattle were shedding L. borgpetersenii; animals were not vaccinated, and the case's serovar was indeterminate. Six wild animals associated with Sites A and C and one environmental sample from Site A were positive for pathogenic Leptospira by PCR. CONCLUSION: These findings highlight the complexity of potential exposures and the difficulty in identifying infection sources for human cases. This reinforces the need for multiple preventive measures such as animal vaccination, the use of personal protective equipment, pest control, and general awareness of leptospirosis to reduce infection risk in agricultural settings. CLINICAL RELEVANCE: Farms with unvaccinated livestock had Leptospira infections, highlighting the importance of animal vaccination. Infections amongst stock that were vaccinated emphasise the importance of best practice vaccination recommendations and pest control. Abbreviations: MAT: Microscopic agglutination test; PIC: Person in charge; PPE: Personalprotective equipment
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    Changing epidemiology of Leptospirosis in New Zealand, with a focus on the novel strain of Leptospira borgpetersenii : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Veterinary Science at Massey University, Palmerston North, New Zealand
    (Massey University, 2024-12-25) Sokolova, Maryna
    In New Zealand, leptospirosis has been a common disease in dairy cattle since the 1940s. Six pathogenic Leptospira serovars from two species have been identified as endemic to New Zealand: Leptospira borgpetersenii serovars (sv.) Hardjobovis (Hardjo), Tarassovi, Ballum, Balcanica, and Leptospira interrogans sv. Pomona and Copenhageni. From these, sv. Pomona and Hardjo are the most commonly reported in cattle, and sv. Ballum, Tarassovi, and Copenhageni are less common. The estimated 99% of the national dairy herd is vaccinated against leptospirosis by vaccines containing antigens to Pomona and Hardjobovis and sometimes vaccines also include Copenhageni antigen. Vaccines for protecting dairy cows against Tarassovi were unavailable in New Zealand before December 2023. Historically, leptospirosis due to Tarassovi infection in New Zealand cattle was considered accidental and clinically unimportant. Serosurveys of apparently healthy cattle in New Zealand showed that over the past fifty years, Tarassovi seroprevalence increased from 6% (50/300) at MAT ≥ 50 to 18% (698/3878) at MAT ≥ 48. More recently, a serology and urine shedding study from 2015- 2016 evaluated the status of 4,000 dairy cows from 200 randomly selected farms, stratified by New Zealand's geographical location and herd size. The study found that on the animal level, 17% of the study animals showed evidence of past infection with Tarassovi, as defined by at least one sample reacting at MAT ≥ 48 for the study's purposes. Moreover, 96% (90/94) of PCR-positive urine samples were sequenced, and 68% (54/80) of those were found to have a novel allele in the sequenced region at the glmU loci. Thus, the use of molecular diagnostic tools, specifically molecular typing targeting a partial region of the glmU gene, allowed New Zealand researchers to identify a novel L. borgpetersenii strain, informally called strain (str.) Pacifica, in the urine of these cows. The 2015-2016 survey reported that dairy cows with Tarassovi titres were associated with urinary shedding, as determined by microscopic agglutination test (MAT) and polymerase chain reaction (PCR) testing, respectively. Because of this association, str. Pacifica is thought to belong to the serogroup Tarassovi. Additionally, the DNA of str. Pacifica has been retrospectively detected in cattle and deer samples dating back as early as 2007. Moreover, the 19-year (1999-2007) average annual incidence of notified human cases of Tarassovi leptospirosis was estimated at 12.59/100,000 in dairy farmers, compared to an overall annual average incidence of 2.01/100,000. These coincidental findings raised public health concerns. Therefore, at least some cases of Tarassovi seropositivity, as identified by MAT and reported before 2021, could partially be attributed to str. Pacifica. To confirm str. Pacifica's serogroup, isolation by culture and complete genetic characterisation of an isolate are required. Since str. Pacifica was only recently detected, its epidemiology, morphology, maintenance, and pathogenicity in the host population, as well as its impact on animal and human health, were not well understood. In this study, we investigated the possibility of isolating str. Pacifica from cow's urine by running a series of laboratory experiments where laboratory-adapted strains were used as a proxy for L. borgpetersenii str. Pacifica in the absence of an isolate to better understand its growth requirements. Laboratory-adapted Leptospira borgpetersenii strains were seeded into different types of media, and Leptospira growth rates were evaluated (Chapter 3). As a result, we ruled out unsuitable media and growth conditions, and this work helped to select the best media and growth conditions for a follow-up field investigation, where freshly collected cow's urine was seeded into selected media. Str. Pacifica was isolated from the urine of a shedding cow using HAN medium at 37°C and 5% CO2. However, this medium failed to sustain str. Pacifica and the culture was lost (Chapter 4). In addition, over the 2020-2021-2022 milking seasons, we collected and tested blood and urine samples from dairy farms, identified as str. Pacifica positive from the 2016 survey. Our results revealed that str. Pacifica was still maintained in the same dairy herds six years after initial detection. Moreover, we reported an estimated prevalence ratio (PR) of 7, indicating that the prevalence of shedders was seven times as high at the beginning than at the end of lactation in primiparous cows (Chapter 4). These findings provide evidence that str. Pacifica is adapted to dairy cows in New Zealand, and the peak shedding in primiparous heifers occurs in early lactation. Since the highest levels of str. Pacifica shedding were detected at the start of the milking season during peak milk production, which also coincides with a relatively short 3-month mating period, the associations between str. Pacifica and milk production and reproductive performance of milking cows were also investigated using herd test data and serological and PCR test results of the 2016 survey. Statistical models, including linear, logistic, and generalised mixed models with fixed and random effects, as well as a shared frailty survival model, were used to evaluate the associations between str. Pacifica positivity and reproduction (Chapter 5) and milk reproduction (Chapter 6) in dairy cows. Results of the statistical analysis of the association between str. Pacifica positivity and reproduction (Chapter 5) of dairy herds showed that str. Pacifica delayed the time from calving to conception (HR = 0.84; 95%: CI 0.74-0.96), although there was no effect on the pregnancy rate (Chapter 5). An analysis of milk production data did not reveal any associations with str. Pacifica at either animal or herd level (Chapter 6). The absence of clinical signs and the lack of association with milk production and reproduction at both the animal and herd levels provides further evidence towards str. Pacifica being well adapted to dairy cows. Therefore, evidence from this thesis suggests that dairy cattle are the maintenance host for str. Pacifica in New Zealand. It is also important to note that str. Pacifica shedders can infect people, especially dairy farmers, milkers, and farm workers who are in regular contact with str. Pacifica-shedding animals. Therefore, the prevention of str. Pacifica transmission via vaccination or the use of appropriate personal protective gear should be prioritised.
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    Molecular typing of Leptospira spp. in farmed and wild mammals reveals new host-serovar associations in New Zealand.
    (Taylor and Francis Group, 2024-01-01) Wilkinson DA; Edwards M; Shum C; Moinet M; Anderson NE; Benschop J; Nisa S
    AIMS: To apply molecular typing to DNA isolated from historical samples to determine Leptospira spp. infecting farmed and wild mammals in New Zealand. MATERIALS AND METHODS: DNA samples used in this study were extracted from urine, serum or kidney samples (or Leptospira spp. cultures isolated from them) collected between 2007 and 2017 from a range of domestic and wildlife mammalian species as part of different research projects at Massey University. Samples were included in the study if they met one of three criteria: samples that tested positive with a lipL32 PCR for pathogenic Leptospira; samples that tested negative by lipL32 PCR but were recorded as positive to PCR for pathogenic Leptospira in the previous studies; or samples that were PCR-negative in all studies but were from animals with positive agglutination titres against serogroup Tarassovi. DNA samples were typed using PCR that targeted either the glmU or gyrB genetic loci. The resulting amplicons were sequenced and typed relative to reference sequences. RESULTS: We identified several associations between mammalian hosts and Leptospira strains/serovars that had not been previously reported in New Zealand. Leptospira borgpetersenii strain Pacifica was found in farmed red deer (Cervus elaphus) samples, L. borgpetersenii serovars Balcanica and Ballum were found in wild red deer samples, Leptospira interrogans serovar Copenhageni was found in stoats (Mustela erminea) and brushtail possums (Trichosurus vulpecula), and L. borgpetersenii was found in a ferret (Mustela putorius furo). Furthermore, we reconfirmed previously described associations including dairy cattle with L. interrogans serovars Copenhageni and Pomona and L. borgpetersenii serovars Ballum, Hardjo type bovis and strain Pacifica, sheep with L. interrogans serovar Pomona and L. borgpetersenii serovar Hardjo type bovis, brushtail possum with L. borgpetersenii serovar Balcanica, farmed deer with L. borgpetersenii serovar Hardjo type bovis and hedgehogs (Erinaceus europaeus) with L. borgpetersenii serovar Ballum. CONCLUSIONS: This study provides an updated summary of host-Leptospira associations in New Zealand and highlights the importance of molecular typing. Furthermore, strain Pacifica, which was first identified as Tarassovi using serological methods in dairy cattle in 2016, has circulated in animal communities since at least 2007 but remained undetected as serology is unable to distinguish the different genotypes. CLINICAL RELEVANCE: To date, leptospirosis in New Zealand has been diagnosed with serological typing, which is deficient in typing all strains in circulation. Molecular methods are necessary to accurately type strains of Leptospira spp. infecting mammals in New Zealand.
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    A cross-sectional investigation of Leptospira at the wildlife-livestock interface in New Zealand
    (PLOS, 2023-09-06) Moinet M; Oosterhof H; Nisa S; Haack N; Wilkinson DA; Aberdein D; Russell JC; Vallée E; Collins-Emerson J; Heuer C; Benschop J; Stevenson B
    There has been a recent upsurge in human cases of leptospirosis in New Zealand, with wildlife a suspected emerging source, but up-to-date knowledge on this topic is lacking. We conducted a cross-sectional study in two farm environments to estimate Leptospira seroprevalence in wildlife and sympatric livestock, PCR/culture prevalence in wildlife, and compare seroprevalence and prevalence between species, sex, and age groups. Traps targeting house mice (Mus musculus), black rats (Rattus rattus), hedgehogs (Erinaceus europaeus) and brushtail possums (Trichosurus vulpecula) were set for 10 trap-nights in March-April 2017 on a dairy (A) and a beef and sheep (B) farm. Trapped wild animals and an age-stratified random sample of domestic animals, namely cattle, sheep and working dogs were blood sampled. Sera were tested by microagglutination test for five serogroups and titres compared using a Proportional Similarity Index (PSI). Wildlife kidneys were sampled for culture and qPCR targeting the lipL32 gene. True prevalence in mice was assessed using occupancy modelling by collating different laboratory results. Infection profiles varied by species, age group and farm. At the MAT cut-point of ≥ 48, up to 78% of wildlife species, and 16-99% of domestic animals were seropositive. Five of nine hedgehogs, 23/105 mice and 1/14 black rats reacted to L. borgpetersenii sv Ballum. The sera of 4/18 possums and 4/9 hedgehogs reacted to L. borgpetersenii sv Hardjobovis whilst 1/18 possums and 1/9 hedgehogs reacted to Tarassovi. In ruminants, seroprevalence for Hardjobovis and Pomona ranged 0-90% and 0-71% depending on the species and age group. Titres against Ballum, Tarassovi and Copenhageni were also observed in 4-20%, 0-25% and 0-21% of domestic species, respectively. The PSI indicated rodents and livestock had the most dissimilar serological responses. Three of nine hedgehogs, 31/105 mice and 2/14 rats were carrying leptospires (PCR and/or culture positive). True prevalence estimated by occupancy modelling in mice was 38% [95% Credible Interval 26, 51%] on Farm A and 22% [11, 40%] on Farm B. In the same environment, exposure to serovars found in wildlife species was commonly detected in livestock. Transmission pathways between and within species should be assessed to help in the development of efficient mitigation strategies against Leptospira.
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    Molecular and eco-epidemiology of Leptospira borgpetersenii serovar Ballum in wild invasive mammals in a farming environment in New Zealand : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Veterinary Science at Massey University, Palmerston North, New Zealand
    (Massey University, 2020) MOINET, Marie
    Leptospirosis is an important zoonosis in New Zealand where it has historically been associated with livestock. Formerly negligible in human cases notified, Leptospira borgpetersenii serovar Ballum—associated with rodents and hedgehogs (Erinaceus europaeus)—is now preponderant. The role of wild introduced mammals in the epidemiology of leptospirosis has been overlooked in New Zealand but remains a critical question. In this thesis, we determined the prevalence of Leptospira serovars, renal colonisation and seroprevalence in wild mammals and sympatric livestock. During a cross-sectional and a longitudinal survey, house mice (Mus musculus), ship rats (Rattus rattus) and hedgehogs were trapped in farms with a history of leptospirosis to collect sera and kidneys. Urine and sera from livestock (dairy or beef cattle, sheep) and dogs were also collected on the same farms. Sera were tested by microagglutination test to identify serovars/serogroups that circulate in wildlife for comparison with those circulating in livestock. Urine and kidney samples were used to determine prevalence by qPCR, to isolate circulating leptospires by culture and subject them to whole genome sequencing, in order to determine their phylogenetic relationships and compare them to other sequences locally, nationally and internationally. Capture-mark recapture (CMR) methods were used to investigate the population dynamics of mice naturally infected with Ballum. Finally, the level of lesions and bacterial load in kidneys were assessed visually by histopathology and put in perspective with other results to investigate reservoir dynamics. Direct or indirect presence of Ballum was found in all wild and domestic species investigated. Overall apparent prevalence in mice, rats and hedgehogs was respectively 46%, 95% CI [39, 52%], 44% [26, 62%] and 27% [11, 50%]. It varied greatly between seasons in mice, with a spring peak (83 to 86%) and minimum in autumn (31 to 37%). Mice densities reached up to 56 mice/ha and varied seasonally in the opposite way, resulting in a relatively constant density of infected mice, ranging 3-8 infected mice/ha. An extremely low rate of mutations hindered the investigation of transmission pathways using genomics. However, despite little or no lesions in all species, the bacterial load was markedly higher in mice, suggesting rats and hedgehogs are secondary hosts. Control strategies to mitigate exposure to Leptospira in NZ should include wild mammals, and especially mice.