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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.

Leptospirosis is a zoonotic disease of global importance. The breadth of Leptospira diversity associated with both human and animal disease poses major logistical challenges to the use of classical diagnostic techniques, and increasingly molecular diagnostic tools are used for their detection. In New Zealand, this has resulted in an increase in positive cases reported nationally that have not been attributed to the infecting serovar or genomospecies. In this study, we used data from all pathogenic Leptospira genomes to identify a partial region of the glmU gene as a suitable locus for the discrimination of the infecting species and serovars of New Zealand-endemic Leptospira. This method can be used in culture and culture-independent scenarios making it flexible for diagnostics in humans, animals, and environmental samples. We explored the use of this locus as a molecular barcoding tool via the Oxford Nanopore Technology (ONT) sequencing platform MinION. Sequences obtained by this method allowed specific identification of Leptospira species in mixed and enriched environmental cultures, however read error inherent in the MinION sequencing system reduced the accuracy of strain/variant identification. Using this approach to characterise Leptospira in enriched environmental cultures, we detected the likely presence of Leptospira genomospecies that have not been reported in New Zealand to date. This included a strain of L. borgpetersenii that has recently been identified in dairy cattle and sequences similar to those of L. mayottensis. L. tipperaryensis, L. dzianensis and L. alstonii., Competing Interests: The authors have declared that no competing interests exist.

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.

Cattle are asymptomatic carriers of Shiga toxin-producing Escherichia coli (STEC) strains that can cause serious illness or death in humans. In New Zealand, contact with cattle feces and living near cattle populations are known risk factors for human STEC infection. Contamination of fresh meat with STEC strains also leads to the potential for rejection of consignments by importing countries. We used a combination of PCR/matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) and whole-genome sequencing (WGS) to evaluate the presence and transmission of STEC on farms and in processing plants to better understand the potential pathways for human exposure and thus mitigate risk. Animal and environmental samples ( n  = 2,580) were collected from six farms and three meat processing plants in New Zealand during multiple sampling sessions in spring of 2015 and 2016. PCR/MALDI-TOF analysis revealed that 6.2% were positive for "Top 7" STEC. Top 7 STEC strains were identified in all sample sources ( n  = 17) tested. A marked increase in Top 7 STEC prevalence was observed between calf hides on farm (6.3% prevalence) and calf hides at processing plants (25.1% prevalence). Whole-genome sequencing was performed on Top 7 STEC bacterial isolates ( n  = 40). Analysis of STEC O26 ( n  = 25 isolates) revealed relatively low genetic diversity on individual farms, consistent with the presence of a resident strain disseminated within the farm environment. Public health efforts should focus on minimizing human contact with fecal material on farms and during handling, transport, and slaughter of calves. Meat processing plants should focus on minimizing cross-contamination between the hides of calves in a cohort during transport, lairage, and slaughter. IMPORTANCE Cattle are asymptomatic carriers of Shiga toxin-producing E. coli (STEC) strains, which can cause serious illness or death in humans. Contact with cattle feces and living near cattle are known risk factors for human STEC infection. This study evaluated STEC carriage in young calves and the farm environment with an in-depth evaluation of six farms and three meat processing plants over 2 years. An advanced molecular detection method and whole-genome sequencing were used to provide a detailed evaluation of the transmission of STEC both within and between farms. The study revealed widespread STEC contamination within the farm environment, but no evidence of recent spread between farms. Contamination of young dairy calf hides increased following transport and holding at meat processing plants. The elimination of STEC in farm environments may be very difficult given the multiple transmission routes; interventions should be targeted at decreasing fecal contamination of calf hides during transport, lairage, and processing., (Copyright © 2021 Browne et al.)

Routinely collected public health surveillance data are often partially complete, yet remain a useful source by which to monitor incidence and track progress during disease intervention. In the 1970s, leptospirosis in New Zealand (NZ) was known as 'dairy farm fever' and the disease was frequently associated with serovars Hardjo and Pomona. To reduce infection, interventions such as vaccination of dairy cattle with these two serovars was implemented. These interventions have been associated with significant reduction in leptospirosis incidence, however, livestock-based occupations continue to predominate notifications. In recent years, diagnosis is increasingly made by nucleic acid detection which currently does not provide serovar information. Serovar information can assist in linking the recognized maintenance host, such as livestock and wildlife, to infecting serovars in human cases which can feed back into the design of intervention strategies. In this study, confirmed and probable leptospirosis notification data from 1 January 1999 to 31 December 2016 were used to build a model to impute the number of cases from different occupational groups based on serovar and month of occurrence. We imputed missing occupation and serovar data within a Bayesian framework assuming a Poisson process for the occurrence of notified cases. The dataset contained 1430 notified cases, of which 927 had a specific occupation (181 dairy farmers, 45 dry stock farmers, 454 meatworkers, 247 other) while the remaining 503 had non-specified occupations. Of the 1430 cases, 1036 had specified serovars (231 Ballum, 460 Hardjo, 249 Pomona, 96 Tarassovi) while the remaining 394 had an unknown serovar. Thus, 47% (674/1430) of observations had both a serovar and a specific occupation. The results show that although all occupations have some degree of under-reporting, dry stock farmers were most strongly affected and were inferred to contribute as many cases as dairy farmers to the burden of disease, despite dairy farmer being recorded much more frequently. Rather than discard records with some missingness, we have illustrated how mathematical modelling can be used to leverage information from these partially complete cases. Our finding provides important evidence for reassessing the current minimal use of animal vaccinations in dry stock. Improving the capture of specific farming type in case report forms is an important next step.

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.

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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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.)

Aims: To describe the common species and the antimicrobial susceptibility of bacteria cultured from samples submitted to commercial veterinary diagnostic laboratories from beef and pre-production dairy cattle between 2003-2016, and to describe the proportion of isolates with multidrug resistance (MDR). Methods: Bacterial culture and antimicrobial susceptibility data from July 2003 to March 2016 were obtained from commercial veterinary diagnostic laboratories in New Zealand. Results were included from samples from beef cattle, irrespective of age or sex, dairy-breed females aged <2 years and dairy-breed males of any age. Submission information provided included the specimen description, the organisms cultured, and the antimicrobial susceptibilities of isolates, if tested. Antimicrobial resistance (AMR) was defined as any isolate not showing susceptibility to an antimicrobial compound and MDR as any isolate showing AMR to ≥3 antimicrobial classes. Results: There were 1,858 unique laboratory submissions, yielding 2,739 isolates. Of these submissions, most were from the Canterbury (389; 21.9%), Manawatu (388; 21.9%) Waikato (231; 12.4%) and Hawke's Bay (136; 7.3%) regions. There were 163 unique species identifications for the 2,739 isolates; the most common were Yersinia pseudotuberculosis (452; 16.5%), Campylobacter jejuni (249; 9.1%), E s cherichia coli (230; 8.4%) and Salmonella enterica serovar Typhimurium (143; 5.2%). Only 251/2,739 (9.2%) isolates from 122/1,858 (6.6%) submissions had antimicrobial susceptibility results. There were no sensitivity results for Yersinia spp., and only one each for Salmonella spp., and Campylobacter spp. Amongst the isolates tested, susceptibility to ampicillin was lowest (33/56; 58.9%). Overall, 57/251 (20.7%) isolates tested for antimicrobial susceptibility had MDR, and MDR was most common for Enterococcus spp. (12/17; 71%) and E. coli (13/30; 43%). Conclusions and Clinical Relevance: This is the first report on antimicrobial susceptibility and MDR in New Zealand beef and pre-production dairy cattle. Findings highlight the limited use of bacterial culture and sensitivity testing by veterinarians and deficits in the information accompanying submissions. A national antimicrobial resistance surveillance strategy that specifically includes this population is recommended.

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.

Background: The Pacific Islands have environmental conditions highly favourable for transmission of leptospirosis, a neglected zoonosis with highest incidence in the tropics, and Oceania in particular. Recent reports confirm the emergence and outbreaks of leptospirosis in the Pacific Islands, but the epidemiology and drivers of transmission of human and animal leptospirosis are poorly documented, especially in the more isolated and less developed islands., Methodology/principal Findings: We conducted a systematic review of human and animal leptospirosis within 25 Pacific Islands (PIs) in Polynesia, Melanesia, Micronesia, as well as Easter Island and Hawaii. We performed a literature search using four international databases for articles published between January 1947 and June 2017. We further included grey literature available on the internet. We identified 148 studies describing leptospirosis epidemiology, but the number of studies varied significantly between PIs. No data were available from four PIs. Human leptospirosis has been reported from 13 PIs, with 63% of all studies conducted in Hawaii, French Polynesia and New Caledonia. Animal leptospirosis has been investigated in 19 PIs and from 14 host species, mainly pigs (18% of studies), cattle (16%) and dogs (11%). Only 13 studies provided information on both human and animal leptospirosis from the same location. Serology results were highly diverse in the region, both in humans and animals., Conclusions/significance: Our study suggests that, as in other tropical regions, leptospirosis is widespread in the PIs while showing some epidemiological heterogeneity. Data are scarce or absent from many PIs. Rodents, cattle, pigs and dogs are all likely to be important carriers, but the relative importance of each animal species in human infection needs to be clarified. Epidemiological surveys with appropriate sampling design, pathogen typing and data analysis are needed to improve our understanding of transmission patterns and to develop effective intervention strategies., Competing Interests: The authors have declared that no competing interests exist.

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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

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., (© 2016 Blackwell Verlag GmbH.)

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., (© 2014 Blackwell Verlag GmbH.)

A study was performed to investigate interlaboratory test agreement between a research and a commercial veterinary diagnostic laboratory on blood and urine samples, and to investigate test agreement between blood, urine, and kidney samples (research laboratory) for leptospirosis diagnosis. Samples were sourced from 399 sheep and 146 beef cattle from a local abattoir. Interlaboratory agreement for real-time quantitative polymerase chain reaction (qPCR) results on urine samples was almost perfect (kappa = 0.90), despite the use of different amplification targets (DNA gyrase subunit B gene vs. 16s ribosomal RNA gene), chemistries (SYTO9 vs. TaqMan probe), and pre-PCR processing. Interlaboratory agreement for microscopic agglutination test (MAT) positivity was almost perfect (kappa = 0.93) for Leptospira borgpetersenii serovar Hardjo subtype Hardjobovis (Hardjobovis) but moderate (kappa = 0.53) for Leptospira interrogans serovar Pomona (Pomona). Among animals that had different titers recorded, higher Hardjobovis and lower Pomona titers were reported by the commercial laboratory than by the research laboratory (P < 0.005). These interlaboratory comparisons can assist researchers and diagnosticians in interpreting the sometimes discrepant test results. Within the research laboratory, the comparison of qPCR results on urine and kidney showed almost perfect agreement (kappa = 0.84), suggesting that the qPCR on these 2 specimens can be used interchangeably. The agreement between MAT positivity and urine and kidney qPCR results was fair (kappa = 0.32 and kappa = 0.33, respectively). However, the prevalence ratio of urine and kidney qPCR positivity in Hardjobovis-seropositive versus Hardjobovis-seronegative sheep indicated that Hardjobovis seropositivity found in sheep may be able to predict shedding or renal carriage., (© 2014 The Author(s).)

A combination of survival and spatial analyses was applied to a dataset containing cattle mortalities on smallholder dairy farms in two separate regions (Tanga and Iringa) of Tanzania. Cattle mortality data for the year 1998 were collected retrospectively during the period January to April 1999. The objectives of the current study were, firstly, to quantify the hazard of mortality and, secondly, to investigate whether there was a spatial pattern in the variation that was unaccounted for in the hazard of mortality. A Cox regression model with farm as a frailty term was used to quantify the hazard of mortality. Geostatistical techniques were used to explore the first- and second-order spatial distribution of the farm frailty term. A total of 1,790 cattle from 400 randomly sampled smallholder dairy farms were included in the study. The overall mortality incidence rate for the study was 11.8 per 100 animal years (95% CI 10.0-13.8). The hazard of mortality for male cattle was 2.4 (95% CI 1.7-3.4) times higher than that of female cattle. There was evidence of a first-order spatial pattern, i.e., a tendency for farms with higher frailty to aggregate.

This article presents the first systematic review of the causes and factors associated with morbidity and mortality, on smallholder dairy farms in Eastern and Southern Africa. It was conducted in August 2007. Primary studies on causes of morbidity and mortality on smallholder dairy farms in the region were identified, examined and descriptive information extracted. Electronic (CAB Abstracts, PubMed, ScienceDirect and Web of Science) and non-electronic databases were used to identify and retrieve the primary studies. The retrieval process included all types of study designs with no restriction on the year of publication and language. Mastitis, tick-borne diseases (TBDs), tick infestation and diarrhoea were among the most commonly documented causes of morbidity. TBDs, diarrhoea and trypanosomiasis were among the most commonly documented causes of mortality; however, a substantial number of mortalities with undiagnosed causes were also reported. Factors ranked as most negatively associated with mastitis were residual calf suckling and increased water availability on the farm while use of common udder towel was the factor most positively associated with mastitis. Zero-grazing was ranked as the factor most negatively associated with TBDs while age was most positively associated. More intervention studies are recommended in the region to better identify animal health constraints and their associated risk factors.