Environmental and sociodemographic factors associated with zoonotic pathogen occurrence in Norway rats (Rattus norvegicus) from Windsor, Ontario.

AIMS: Rat-associated zoonotic pathogen transmission at the human-wildlife interface is a public health concern in urban environments where Norway rats (Rattus norvegicus) thrive on abundant anthropogenic resources and live in close contact with humans and other animal species. To identify potential factors influencing zoonotic pathogen occurrence in rats, we investigated associations between environmental and sociodemographic factors and Leptospira interrogans and Bartonella spp. infections in rats from Windsor, Ontario, Canada, while controlling for the potential confounding effects of animal characteristics (i.e., sexual maturity and body condition). METHODS AND RESULTS: Between November 2018 and June 2021, 252 rats were submitted by collaborating pest control professionals. Kidney and spleen samples were collected for L. interrogans and Bartonella spp. PCR and sequencing, respectively. Of the rats tested by PCR, 12.7% (32/252) were positive for L. interrogans and 16.3% (37/227) were positive for Bartonella species. Associations between infection status and environmental and sociodemographic variables of interest were assessed via mixed multivariable logistic regression models with a random intercept for social group and fixed effects to control for sexual maturity and body condition in each model. The odds of L. interrogans infection were significantly higher in rats from areas with high building density (odds ratio [OR]: 3.76; 95% CI: 1.31-10.79; p = 0.014), high human population density (OR: 3.31; 95% CI: 1.20-9.11; p = 0.021), high proportion of buildings built in 1960 or before (OR: 11.21; 95% CI: 2.06-60.89; p = 0.005), and a moderate number of reports of uncollected garbage compared to a low number of reports (OR: 4.88; 95% CI: 1.01-23.63; p = 0.049). A negative association was observed between median household income and Bartonella spp. infection in rats (OR: 0.26; 95% CI: 0.08-0.89; p = 0.031). CONCLUSIONS: Due to the complexity of the ecology of rat-associated zoonoses, consideration of environmental and sociodemographic factors is of critical importance to better understand the nuances of host-pathogen systems and inform how urban rat surveillance and intervention efforts should be distributed within cities.

A Multi-landscape Assessment of Leptospira Prevalence on a Diversity of Small Mammals.

Leptospirosis is a major zoonotic disease, especially in the tropics, and rodents were known to be carriers of this bacterium. There was established information on Leptospira prevalence among animal reservoirs in human-dominated landscapes from previous literature. However, there was very little focus given comparing the prevalence of Leptospira in a wide range of habitats. An extensive sampling of small mammals from various landscapes was carried out, covering oil palm plantations, paddy fields, recreational forests, semi-urbans, and wet markets in Peninsular Malaysia. This study aims to determine the prevalence of pathogenic Leptospira in a diversity of small mammals across different landscapes. Cage-trapping was deployed for small mammals' trappings, and the kidneys of captured individuals were extracted, for screening of pathogenic Leptospira by polymerase chain reaction (PCR) using LipL32 primer. Eight microhabitat parameters were measured at each study site. Out of 357 individuals captured, 21 (5.9%) were positive for pathogenic Leptospira of which recreational forest had the highest prevalence (8.8%) for landscape types, whereas Sundamys muelleri shows the highest prevalence (50%) among small mammals' species. Microhabitat analysis reveals that rubbish quantity (p < 0.05) significantly influenced the Leptospira prevalence among small mammals. Furthermore, nMDS analysis indicates that the presence of faeces, food waste, and exposure to humans in each landscape type also were linked with high prevalence of pathogenic Leptospira among the small mammals. This study supplements previous studies on pathogenic Leptospira prevalence across different landscape types, and the major microhabitat factors associated with Leptospira prevalence. This information is crucial for epidemiological surveillance and habitat management to curb the possibility of the disease outbreaks.

Development and evaluation of polyclonal antibody-based antigen detection ELISA and dot blot assays as a less costly diagnostic alternative for pathogenic Leptospira infection.

Leptospirosis is an infectious, zoonotic disease of worldwide distribution, the cause of which is infection by pathogenic Leptospira. In Chile, dairy cattle are recognized a significant source in the maintenance and transmission of this infection, which causes economic losses and represents an infection threat to workers in the dairy industry. The infection is underestimated in cattle, due to the lack of clinical, pathognomonic signs, as well as the low efficiency of current diagnostic techniques. In this study, we developed antigen ELISA and dot blot assays, based on polyclonal antibodies, to detect pathogenic Leptospira in the urine samples of dairy cattle. The proposed tests showed an acceptable diagnostic accuracy, based on an analytical sensitivity of 1·104 Leptospira per mL for ELISA, and 3.2·103 for dot blot. These results corresponded with those obtained by qPCR, and the use of urine samples allowed us to propose new diagnostic alternatives for pathogenic Leptospira infection at a low cost, which can provide information on active infection status, which is a key element in control programs both at individual and herd level.

Impact of cattle on joint dynamics and disease burden of Japanese encephalitis and leptospirosis.

Japanese encephalitis (JE) is a mosquito-borne neglected tropical disease. JE is mostly found in rural areas where people usually keep cattle at home for their needs. Cattle in households reduce JE virus infections since they distract vectors and act as a dead-end host for the virus. However, the presence of cattle introduces risk of leptospirosis infections in humans. Leptospirosis is a bacterial disease that spreads through direct or indirect contact of urine of the infected cattle. Thus, cattle have both positive and negative impacts on human disease burden. This study uses a mathematical model to study the joint dynamics of these two diseases in the presence of cattle and to identify the net impact of cattle on the annual disease burden in JE-prevalent areas. Analysis indicates that the presence of cattle helps to reduce the overall disease burden in JE-prevalent areas. However, this reduction is dominated by the vector's feeding pattern. To the best of our knowledge, this is the first study to examine the joint dynamics of JE and leptospirosis.

Estimation of the burden of leptospirosis in New Zealand.

BACKGROUND: Human leptospirosis mainly affects people in close occupational contact with domestic livestock and their products in New Zealand. The disease has an unquantified impact on both human health and animal production in the country. This study aimed to estimate the burden of leptospirosis in terms of disability-adjusted life years (DALYs) and cost associated with loss due to absence from work, treatment of disease, animal production loss and cost of vaccination. METHODS: Previously published studies of abattoir workers farmers, and veterinarians, reporting annual risks of influenza-like illness attributable to Leptospira infection, were used to estimate the expected number of cases in a year. The cost of lost animal production was based on results of observational studies in beef cattle, sheep and deer conducted in New Zealand. RESULTS: Expected median annual number of severe and mild cases of human leptospirosis was 2,025 (95% probability interval [95% PI] 1,138-3,422). Median annual DALYs were 0.42 (95% PI: 0.06-2.40) per 100,000 people for the entire population, and 15.82 (95% PI: 2.09-90.80) per 100,000 people working in at-risk occupations (i.e. abattoir workers, farmers and veterinarians). Human infection resulted in a median cost of 4.42 (95% PI: 2.04-8.62) million US dollars (USD) due to absence from work and disease treatment. Median production loss cost in beef cattle, sheep and deer was USD 7.92 (95% PI: 3.75-15.48) million, while median vaccination cost in cattle, (including dairy), sheep and deer was USD 6.15 (95% PI: 5.30-7.03) million. Total annual cost of leptospirosis plus vaccination was USD 18.80 (95% PI: 13.47-27.15) million, equivalent to USD 440,000 (95% PI: 320,000-640,000) per 100,000 people. CONCLUSION: This study provides an estimate of the disease burden and cost of leptospirosis in New Zealand that could support occupational health authorities and livestock industries in assessing interventions for this disease.

Assessment of the immunogenicity of the leptospiral LipL32, LigAni, and LigBrep recombinant proteins in the sheep model.

Veterinary leptospirosis vaccines are composed of bacterins and present limitations, for example, the need for bacteriological culture and serovar-dependent immunity. Recombinant antigens represent a promising alternative. LigAni, LigBrep, and LipL32 proteins have been shown to promote a protective immune response against the homologous challenge in hamsters. Therefore, the next step is to evaluate the immunological properties of these immunogens in the actual hosts, as ruminants, which has never been performed before. The objective of this study was to evaluate the immunogenicity and potential adverse effects of the recombinant proteins LigAni, LigBrep, and LipL32 in the ovine model. For this, 16 Santa Inês sheep were allocated into three groups: two experimental (Groups A and B) and one control group (Group C). Group A was inoculated with a formulation containing the recombinant proteins in combination with the aluminum hydroxide adjuvant; Group B was inoculated with a formulation containing the recombinant proteins in combination with the Montanide adjuvant; and Group C was inoculated with adjuvants only. The results revealed that formulations containing the recombinant proteins induced total IgG seroconversion and led to a significant increase in antibody titers in the sheep model. Besides, there were no clinical changes or adverse effects. Thus, LigAni, LigBrep, and LipL32 proteins elicited a significant humoral immune response with elevated serum IgG levels, demonstrating that they possess the immunogenic and safety characteristics necessary to sustain their potential use as leptospirosis vaccines in the ruminant model.

Chronology of emergence of the genus Leptospira and over-representation of gene families enriched by vitamin B2, B12 biosynthesis, cell adhesion and external encapsulating structure in L. interrogans isolates from asymptomatic dogs.

The spirochete species Leptospira interrogans is the most common cause of leptospirosis, producing acute to chronic infections in most mammalian species. This pathogenic bacterium has an ability to evolve in many ways to occupy various environments and hosts. In this study, we performed chronology analysis to look for insight into the emergence of Leptospira species, focusing on L. interrogans, and investigated gene gain and loss related to their adaptation in strains isolated from asymptomatic dogs. Chronology analysis revealed that the emergence of L. interrogans was around 53.7 million years ago (MYA), corresponding to the Paleogene period that coincided with an optimal climatic temperature and the evolution of suitable mammalian hosts. Gene families encoding for vitamin B2, B12 biosynthesis, cell adhesion and external encapsulating structure were found to be enriched in L. interrogans isolated from the urine of asymptomatic dogs. The activity of these gene families may have favored adaptations resulting in chronic infections.

Pharmacokinetics of enrofloxacin HCl-2H2 O (ENRO-C), PK/PD, and Monte Carlo modeling vs. Leptospira spp. in cows.

The pharmacokinetics, PK/PD ratios, and Monte Carlo modeling of enrofloxacin HCl-2H2 O (Enro-C) and its reference preparation (Enro-R) were determined in cows. Fifty-four Jersey cows were randomly assigned to six groups receiving a single IM dose of 10, 15, or 20 mg/kg of Enro-C (Enro-C10 , Enro-C15 , Enro-C20 ) or Enro-R. Serial serum samples were collected and enrofloxacin concentrations quantified. A composite set of minimum inhibitory concentrations (MIC) of Leptospira spp. was utilized to calculate PK/PD ratios: maximum serum concentration/MIC (Cmax /MIC90 ) and area under the serum vs. time concentration of enrofloxacin/MIC (AUC0-24 /MIC90 ). Monte Carlo simulations targeted Cmax /MIC = 10 and AUC0-24 /MIC = 125. Mean Cmax obtained were 6.17 and 2.46 µg/ml; 8.75 and 3.54 µg/ml; and 13.89 and 4.25 µg/ml, respectively for Enro-C and Enro-R. Cmax /MIC90 ratios were 6.17 and 2.46, 8.75 and 3.54, and 13.89 and 4.25 for Enro-C and Enro-R, respectively. Monte Carlo simulations based on Cmax /MIC90  = 10 indicate that only Enro-C15 and Enro-C20 may be useful to treat leptospirosis in cows, predicting a success rate ≥95% when MIC50  = 0.5 µg/ml, and ≥80% when MIC90  = 1.0 µg/ml. Although Enro-C15 and Enro-C20 may be useful to treat leptospirosis in cattle, clinical trials are necessary to confirm this proposal.

Human leptospirosis in Seychelles: A prospective study confirms the heavy burden of the disease but suggests that rats are not the main reservoir.

BACKGROUND: Leptospirosis is a bacterial zoonosis caused by pathogenic Leptospira for which rats are considered as the main reservoir. Disease incidence is higher in tropical countries, especially in insular ecosystems. Our objectives were to determine the current burden of leptospirosis in Seychelles, a country ranking first worldwide according to historical data, to establish epidemiological links between animal reservoirs and human disease, and to identify drivers of transmission. METHODS: A total of 223 patients with acute febrile symptoms of unknown origin were enrolled in a 12-months prospective study and tested for leptospirosis through real-time PCR, IgM ELISA and MAT. In addition, 739 rats trapped throughout the main island were investigated for Leptospira renal carriage. All molecularly confirmed positive samples were further genotyped. RESULTS: A total of 51 patients fulfilled the biological criteria of acute leptospirosis, corresponding to an annual incidence of 54.6 (95% CI 40.7-71.8) per 100,000 inhabitants. Leptospira carriage in Rattus spp. was overall low (7.7%) but dramatically higher in Rattus norvegicus (52.9%) than in Rattus rattus (4.4%). Leptospira interrogans was the only detected species in both humans and rats, and was represented by three distinct Sequence Types (STs). Two were novel STs identified in two thirds of acute human cases while noteworthily absent from rats. CONCLUSIONS: This study shows that human leptospirosis still represents a heavy disease burden in Seychelles. Genotype data suggests that rats are actually not the main reservoir for human disease. We highlight a rather limited efficacy of preventive measures so far implemented in Seychelles. This could result from ineffective control measures of excreting animal populations, possibly due to a misidentification of the main contaminating reservoir(s). Altogether, presented data stimulate the exploration of alternative reservoir animal hosts.

Colony-level assessment of Brucella and Leptospira in the Guadalupe fur seal, Isla Guadalupe, Mexico.

The relatively small population size and restricted distribution of the Guadalupe fur seal Arctocephalus townsendi could make it highly vulnerable to infectious diseases. We performed a colony-level assessment in this species of the prevalence and presence of Brucella spp. and Leptospira spp., pathogenic bacteria that have been reported in several pinniped species worldwide. Forty-six serum samples were collected in 2014 from pups at Isla Guadalupe, the only place where the species effectively reproduces. Samples were tested for Brucella using 3 consecutive serological tests, and for Leptospira using the microscopic agglutination test. For each bacterium, a Bayesian approach was used to estimate prevalence to exposure, and an epidemiological model was used to test the null hypothesis that the bacterium was present in the colony. No serum sample tested positive for Brucella, and the statistical analyses concluded that the colony was bacterium-free with a 96.3% confidence level. However, a Brucella surveillance program would be highly recommendable. Twelve samples were positive (titers 1:50) to 1 or more serovars of Leptospira. The prevalence was calculated at 27.1% (95% credible interval: 15.6-40.3%), and the posterior analyses indicated that the colony was not Leptospira-free with a 100% confidence level. Serovars Icterohaemorrhagiae, Canicola, and Bratislava were detected, but only further research can unveil whether they affect the fur seal population.

Effect of exposure to Neospora caninum, Salmonella, and Leptospira interrogans serovar Hardjo on the economic performance of Irish dairy herds.

The objective of the current study was to quantify the effects of exposure to Salmonella, Neospora caninum, and Leptospira interrogans serovar Hardjo (L. hardjo) on dairy farm profitability and to simulate the effect of vaccination for Salmonella and L. hardjo on dairy farm profitability. The production effects associated with exposure to each of these pathogens in study herds were defined under 3 categories: (1) milk production effects, (2) reproduction effects (including culling), and (3) mortality effects. The production effects associated with exposure to Salmonella, N. caninum, and L. hardjo were incorporated into the Moorepark Dairy Systems Model. In the analysis, herds negative for exposure to Salmonella, N. caninum, and L. hardjo were assumed baseline herds, with all results presented relative to this base. In simulations examining the effect of vaccination for Salmonella and L. hardjo on farm profitability, vaccinated herds (vaccination costs included) were considered as baseline herds and results were presented relative to this base. Total annual profits in unvaccinated herds were reduced by €77.31, €94.71, and €112.11 per cow at milk prices of €0.24, €0.29, and €0.34/L, respectively, as a result of exposure to Salmonella. In the current study, herds positive for exposure to Salmonella recorded a 316-kg reduction in milk yield, whereas no association was detected between exposure to N. caninum or L. hardjo and milk production. Exposure to both N. caninum and L. hardjo was associated with compromised reproductive performance. Herds positive for exposure to N. caninum and Salmonella had greater rates of adult cow mortality and calf mortality, respectively. Vaccination for both Salmonella and L. hardjo was associated with improved performance in study herds. Exposure to N. caninum resulted in a reduction in annual farm profits of €11.55, €12, and €12.44 per cow at each milk price, whereas exposure to L. hardjo resulted in a reduction in annual farm profits of €13.83, €13.78, and €13.72 per cow at each milk price. Herds that tested positive for exposure to Salmonella and L. hardjo were compared with herds vaccinated for the respective pathogens. Herds vaccinated for Salmonella generated €67.09, €84.48, and €101.89 per cow more profit at each milk price compared with herds positive for exposure. Similarly, herds vaccinated for L. hardjo generated €9.74, €9.69, and €9.63 per cow more profit compared with unvaccinated exposed herds. However, herds that tested negative for exposure to Salmonella and L. hardjo generated additional profits of €10.22 and €4.09 per cow, respectively, compared with vaccinated baseline herds.

An assessment of zoonotic and production limiting pathogens in rusa deer (Cervus timorensis rusa) from Mauritius.

A population of approximately 70,000 rusa deer (Cervus timorensis russa) represents the most important mammal species reared for food on the island of Mauritius, being the main source of red meat for the local population. However, very limited information is available on the circulation of pathogens affecting the productivity and health of this species. To produce baseline data on the circulation of infectious pathogens in rusa deer under production, a serological survey and/or direct pathogen detection for six selected infectious diseases was undertaken in 2007 in a sample of 53% of the herds reared in semi-free-ranging conditions in hunting estates. Seropositive results were recorded for Johne's disease with an indirect ELISA test (1.7%, n = 351), heartwater with an immunofluorescence antibody test (IFAT) (95.5%, n = 178) and leptospirosis with a Microscopic Agglutination Test (MAT) (25.9%, n = 363). Significant associations were found between seroprevalence to some of the leptospiral serogroups detected (Tarassovi, Pomona, Sejroe and Mini) and age of the animals, animal density or location of the estates (being more prevalent in hotter and more humid areas). In addition, Mycobacterium bovis and M. avium subspecies paratuberculosis were confirmed in two deer carcasses by culture and PCR, respectively. No antibodies against Brucella spp. nor Rift Valley Fever virus were detected with the use of respective indirect ELISA's. The results obtained suggest that the population of rusa deer from Mauritius is exposed to a wide range of pathogens which may affect their productivity. In addition, the results highlight the potential public health risks incurred by deer industry workers and consumers. This survey fills an important gap in knowledge regarding the health of tropical deer meat in Mauritius and justifies the need to implement more regular surveys of selected pathogens in the deer population.

[Leptospirosis outbreak in calves from Corrientes Province, Argentina]. / Brote de leptospirosis en terneros en recría en la provincia de Corrientes, Argentina.

Leptospirosis is an infectious disease resulting in significant economic losses in livestock production. This disease causes abortion, embryo death, death of calves within the first few days of life and mastitis. We report a leptospirosis outbreak in calf growing and fattening. Histopathological and hemoparasite studies, immunofluorescence, and bacterial cultures were performed. A strain of Leptospira interrogans serovar Pomona was isolated from samples collected from dead calves.

Assessment of exposure to Leptospira serovars in veterinary staff and dog owners in contact with infected dogs.

OBJECTIVE: To assess patterns of seroreactivity to Leptospira serovars in veterinary professional staff and dog owners exposed to dogs with acute leptospirosis and to contrast these patterns in people with those observed in dogs. DESIGN: Cross-sectional study. SAMPLE POPULATION: Human subjects consisted of 91 people (50 veterinarians, 19 technical staff, 9 administrative personnel, and 13 dog owners) exposed to dogs with leptospirosis. Canine subjects consisted of 52 dogs with naturally occurring leptospirosis admitted to the University of Bern Vetsuisse Faculty Small Animal Clinic in 2007 and 2008. PROCEDURES: People were tested for seroreactivity to regionally prevalent Leptospira serovars by use of a complement fixation test. A questionnaire designed to identify risk factors associated with seropositivity was used to collect demographic information from each study participant. Dogs were tested for seroreactivity to Leptospira serovars by use of a microscopic agglutination test. RESULTS: On the basis of microscopic agglutination test results, infected dogs were seropositive for antibodies against Leptospira serovars as follows (in descending order): Bratislava (43/52 [83%]), Australis (43/52 [83%]), Grippotyphosa (18/52 [35%]), Pomona (12/52 [23%]), Autumnalis (6/52 [12%]), Icterohemorrhagiae (4/52 [8%]), Tarassovi (2/52 [4%]), and Canicola (1/52 [2%]). All 91 people were seronegative for antibodies against Leptospira serovars. Therefore, statistical evaluation of risk factors and comparison of patterns of seroreactivity to Leptospira serovars between human and canine subjects were limited to theoretical risks. CONCLUSIONS AND CLINICAL RELEVANCE: Seroreactivity to Leptospira serovars among veterinary staff adhering to standard hygiene protocols and pet owners exposed to dogs with acute leptospirosis was uncommon.

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.

Global change: impact, management, risk approach and health measures--the case of Europe.

Global changes, including an increase in trade and global warming, which act on the environment, are likely to impact on the evolution of pathogens and hence of diseases. To anticipate the risks created by this new situation, a French group of experts has developed a method for prioritising animal health risks. This is a two-phase method: the first step is to identify the diseases whose incidence or geographical distribution could be affected by the changes taking place, and the second step is to evaluate the risk of each of these diseases. As a result of this process, six priority diseases were selected: bluetongue, Rift Valley fever, West Nile fever, visceral leishmaniasis, leptospirosis and African horse sickness. The main recommendations were: to develop epidemiological surveillance, to increase knowledge of epidemiological cycles, to develop research into these diseases and to pool cross-border efforts to control them.

A serologic assessment of exposure to viral pathogens and Leptospira in an urban raccoon (Procyon lotor) population inhabiting a large zoological park.

In urban environments, raccoons (Procyon lotor) may act as reservoirs for an array of pathogenic organisms, presenting spillover risks for human, domestic animal, and captive (zoo) animal populations. Over 5 yr, 159 raccoons from a high-density raccoon population in St. Louis, Missouri (USA), were surveyed for exposure to canine distemper virus (CDV), canine adenovirus 1 (CAV-1); feline parvovirus (FPV; =feline panleukopenia), and several serovars of Leptospira interrogans. Exposure to each of the viruses and two Leptospira serovars (grippotyphosa and icterohemorrhagiae) was detected (prevalence of CDV = 54.1%; FPV = 49.7%; CAV-1 = 6.9%; L. interrogans icterohemorrhagiae = 8.9%; L. interrogans grippotyphosa = 6.3%). Eighty percent of raccoons showed evidence of exposure to at least one of the five primary pathogens, and 39% were positive for multiple species. Among the viruses, there was a significant co-occurrence of CDV and CAV-1. Longitudinal data on a subset of animals revealed that among individuals who were diagnosed as seropositive on first capture, 33-100% became seronegative for the pathogen of interest when reexamined at a later date. Thus, free-ranging urban raccoons have been exposed to multiple infectious agents, some of which may pose risks to humans and to nonvaccinated domestic and captive animal populations.

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

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