16例犬钩端螺旋体病例的回顾性分析

为了解犬钩端螺旋体病的临床特点和诊疗方法,对2017年10月至2018年7月在浙江大学动物医院确诊的16例钩端螺旋体病犬的发病和诊疗情况进行了回顾分析。PCR诊断钩端螺旋体免疫的公犬是高发群体,雨水较多的月份发病率高于雨水少的月份,由于周边地区金毛、贵宾和田园犬的保有量相对其他品种要高,故在患犬中占比较高。病初主要表现为呕吐、脱水、精神委靡、结膜潮红、食欲废绝等症状。临床检查发现血尿素氮、血肌酐、血磷等指标明显升高,白细胞不同程度升高,部分病例出现贫血。B超检查可见肾肿大、肾实质损伤。主要酸碱紊乱类型为代谢性酸中毒和呼吸性酸中毒,低钠血症最为常见,并发症有肝损伤、胰腺炎、肺炎等。     

Immunological and molecular characterization of Leptospira interrogans isolated from a bovine foetus

Cattle are commonly infected with pathogenic leptospires, and similarly to rodents, they excrete the bacteria in their urine and can transmit the pathogen from animal to animal or animal to human. Thus, surveillance and monitoring systems for detection of new Leptospira serovars are important for the control of leptospirosis. Here, we report the isolation of a spirochete from a stillborn bovine foetus and its characterization by immunological and molecular techniques. A variable number tandem repeat profile using seven discriminatory primers identified the spirochete as belonging to species Leptospira interrogans serogroup Australis serovar Muenchen. A phenotypic analysis using monoclonal antibodies (mAbs) against leptospiral membrane-associated proteins confirmed the expression of important virulence and pathogenicity factors (LipL32 and LigBrep). Out of 120 reference sera tested, 22 positive (36.66%) and 9 negative (15%) also reacted with the new isolate. Furthermore, the serovar Muenchen isolate was virulent in hamster model. The animal inoculated developed acute lethal infection characterized by hepatic, pulmonary and renal lesions. Local isolates exhibited unique characteristics that differed from those of reference strains; therefore, isolation of leptospires is useful in the surveillance of local pathogenic serovars. In conclusion, the data obtained from this study can contribute to the epidemiological understanding and control of leptospirosis in southern Brazil.     

Seasonality of canine leptospirosis in the United States and Canada and its association with rainfall

A retrospective study was undertaken to describe seasonal patterns of leptospirosis diagnosed at veterinary teaching hospitals in the United States and Canada, and to determine if occurrence of cases (diagnoses) was associated with rainfall. The veterinary medicine database (VMDB) was searched for records of dogs in which a diagnosis of leptospirosis was made, and average monthly rainfall recorded at the city where each hospital is located was calculated from historical data. Time-series analysis was used to identify the variables that best described occurrence of cases of leptospirosis. Three-hundred and forty dogs were diagnosed with leptospirosis at 22 veterinary teaching hospitals between 1983 and 1998. Most cases (n = 184) were diagnosed between August and November during each year of the study. A significant correlation (r = 0.41) was detected between the number of cases of leptospirosis diagnosed and average rainfall recorded 3-months prior to diagnosis. The best-fitting (Akaike's corrected information criterion = 2.01) regression on the leptospirosis case series included cases diagnosed in the previous month and 12-months previously, and rainfall recorded 3 months previously. Leptospirosis has a seasonal distribution (late summer to fall), and rainfall can be used to predict the occurrence of leptospirosis. Leptospirosis can affect the health of dogs seriously and its prevalence is increasing, but the disease is preventable by vaccination.     

Epidemiology of an outbreak of leptospirosis in man and dog

An outbreak of leptospirosis in man and dog occurred during the monsoon in Madras city, India. 48 (50.5%) of the 95 human sera tested were positive for leptospirosis. Of these, 32 showed the presence of agglutinin to serovar icterohaemorrhagiae. 20 (21.3%) of the 94 canine sera tested were positive for leptospirosis. Of these, 10 had agglutinins to serovar icterohaemorrhagiae and 9 to serovar canicola. Among murine reservoirs, 8 (25%) of 32 rat sera and 10 (41.6%) of 24 bandicoot sera were positive. Bandicoot and rat sera contained 70 and 50% of agglutinins to serovar icterohaemorrhagiae, respectively. Urine culture lead to isolate a icterohaemorrhagiae strain from a patient and a canicola one from a dog. The changing trends of epidemiology in the transmission of leptospirosis to man and dog has been discussed.     

Evaluation of environmental risk factors for leptospirosis in dogs: 36 cases (1997-2002)

OBJECTIVE: To identify environmental risk factors for leptospirosis. DESIGN: Retrospective study. ANIMALS: 36 dogs with leptospirosis and 138 dogs seronegative for leptospirosis as determined by microscopic agglutination test for antibodies against Leptospira spp. PROCEDURES: Medical records of dogs evaluated for leptospirosis from 1997 though 2002 were identified. Owner address was used to geocode locations of dogs, and location-specific environmental risk factor data were obtained by use of a geographic information system. Risk of leptospirosis was estimated by odds ratios, controlling for potential confounding by dog age, sex, and breed. RESULTS: Leptospirosis in 19 of the 30 dogs in which an infecting Leptospira serovar could be identified was associated with Leptospira kirschneri serovar grippotyphosa infection. Dogs in which a diagnosis of leptospirosis was made, and dogs with leptospirosis caused by L kirschneri serovar grippotyphosa, were more likely to have addresses located in areas classified as rural in 1990 but urban in 2000. By use of information on recent urbanization and a logistic regression model, the status of 81.6% and 89.8% of dogs with leptospirosis and leptospirosis caused by serovar grippotyphosa, respectively, were correctly classified. Other environmental variables (proximity to streams, recreational areas, farmland, wetlands, areas subject to flooding, and areas with poor drainage; annual rainfall; and county cattle or pig population) did not significantly improve accuracy of classification. CONCLUSIONS AND CLINICAL RELEVANCE: Dogs in periurban areas are at greater risk of leptospirosis. Vaccination of dogs in these areas to protect against leptospirosis should be considered.     

2010 ACVIM small animal consensus statement on leptospirosis: diagnosis, epidemiology, treatment, and prevention

This report offers a consensus opinion on the diagnosis, epidemiology, treatment, and prevention of leptospirosis in dogs, an important zoonosis. Clinical signs of leptospirosis in dogs relate to development of renal disease, hepatic disease, uveitis, and pulmonary hemorrhage. Disease may follow periods of high rainfall, and can occur in dogs roaming in proximity to water sources, farm animals, or wildlife, or dogs residing in suburban environments. Diagnosis is based on acute and convalescent phase antibody titers by the microscopic agglutination test (MAT), with or without use of polymerase chain reaction assays. There is considerable interlaboratory variation in MAT results, and the MAT does not accurately predict the infecting serogroup. The recommended treatment for optimal clearance of the organism from renal tubules is doxycycline, 5 mg/kg p.o. q12h, for 14 days. Annual vaccination can prevent leptospirosis caused by serovars included in the vaccine and is recommended for dogs at risk of infection.     

Increase in seroprevalence of canine leptospirosis and its risk factors,Ontario 1998–2006

Canine leptospirosis has been described as having re-emerged in North America around the mid-1990s, with a change in the epidemiology of the infecting serovars responsible for the disease emergence. A retrospective case-control study was conducted to examine the re-emergence of seroprevalent cases of canine leptospirosis in Ontario using serology submission records from 1406 dogs from January 1, 1998 to December 31, 2006. The data collected [results of the microscopic agglutination test (MAT), veterinary clinic postal code, age, sex, neutering status, and breed] were analyzed by multivariable logistic regression, generalized linear mixed modeling, and Cochran-Armitage test for trends in proportions. Dogs in urban areas appeared to be at significantly higher risk than dogs in rural areas for the entire study period [odds ratio (OR) = 1.6, confidence interval (CI) = 1.2–2.3], though this was not as marked as in other studies. Results indicated that canine leptospirosis in Ontario is a disease of all breeds and ages, regardless of gender. No geographic clustering was noted, but clustering of cases by clinic within geographic areas suggested differences in awareness or in diagnosis by veterinarians. A distinctive seasonal pattern of leptospirosis, with more cases occurring during the summer and fall, as found in previous studies, was also observed in this study. The temporal trend analysis was consistent with an increasing proportion or re-emergence of seroprevalent cases of canine leptospirosis since 1998, suggesting that the putative increase in canine leptospirosis has been genuine.     

Detection of anti-borna disease virus antibodies from cats in Asian countries, Japan, Philippines and Indonesia using electrochemiluminescence immunoassay

Anti-Borna disease virus (BDV) antibodies were detected from cats in Japan, Philippines and Indonesia by using electrochemiluminescence immunoassay. Positive rates were 3.1%, 3.8% and 2.0% in Japan, Philippines and Indonesia, respectively. There was no differences in the positive rate of anti-BDV antibodies between male and female cats and among habitats. While, a significantly (P<0.05) higher prevalence (6.5%) was found in the oldest age group (more than 6 years) cats.     

Reproductive and leptospiral studies on beef cattle in central Queensland

The effect of rainfall on reproductive performance in beef cattle and the effects of rainfall and soil type on the prevalence of leptospirosis in beef cattle in inland central Queensland are described. Low annual rainfall produced a significant (P less than 0.05) decrease in calf branding rates. Leptospirosis (due to serovar hardjo) was serologically more prevalent after rain and on farms with high water holding capacity soils but there was no significant difference in branding rates between cattle on high or low water holding capacity soils. As a secondary mid-year rainfall peak is a feature of the area, leptospirosis due to serovar hardjo will tend to spread when most of the breeding herd is in the last trimester of pregnancy. The prevalence of leptospirosis due to serovar pomona is significantly lower in the region.     

Treatment and outcome of dogs with leptospirosis: 36 cases (1990-1998)

OBJECTIVE: To characterize serologic and clinical features and outcome of dogs with leptospirosis that were treated conservatively (i.e., medical management alone) or with hemodialysis. DESIGN: Retrospective study. ANIMALS: 36 dogs with leptospirosis. PROCEDURE: History; results of physical examinations, ultrasonography, and serologic, hematologic, and serum biochemical analyses; time to resolution of azotemia; and outcome were obtained from medical records. Dogs were treated conservatively (n = 22) or with hemodialysis (14). RESULTS: Between 1990 and 1998, amount of rainfall was positively correlated with number of cases of leptospirosis identified per year. Serum antibodies against 6 Leptospira serovars were measured, and titers were highest to Leptospira pomona in 16 (44%) dogs, L bratislava in 9 (25%) dogs, and L hardjo in 1 (3%) dog. Eight (22%) dogs had equally high titers to L pomona and L bratislava, 1 (3%) had equally high titers to L grippotyphosa and L canicola, and 1 (3%) had high titers to L grippotyphosa, L pomona, L canicola, and L bratislava. During initial evaluation, all dogs were azotemic. Thirty (83%) dogs survived, including 12 of 14 (86%) dogs treated with hemodialysis and 18 of 22 (82%) treated conservatively. Serum creatinine concentration was similar in both groups after resolution of clinical signs. CONCLUSIONS AND CLINICAL RELEVANCE: Infection with L pomona and L bratislava was recognized as a cause of leptospirosis in dogs, and resulted in development of acute renal failure with various degrees of azotemia. Prognosis for dogs with mild to moderate azotemia was good with conservative treatment, whereas treatment with hemodialysis appeared to improve prognosis for dogs with severe azotemia.     

New enzyme-linked immunoassay for the detection of specific antibodies against multiple Leptospira serogroups in bovine sera

Leptospirosis is a zoonotic disease with worldwide endemicity in Argentina, it is a significant public health problem in low-income populations. Bovine leptospirosis is a serious economic problem for cattle production, causing abortions, reduced milk yield, mortality in calves and decreased daily weight gain. We developed an enzyme-linked immunosorbent assay (ELISA) with sonicated Leptospira interrogans serogroup Icterohaemorrhagiae serovar Copenhageni M 20. We evaluated its performance for the detection of specific antibodies against multiple Leptospira serogroups in bovine. The microscopic agglutination test (MAT) was used as the gold standard. The performance of this ELISA was evaluated with a panel of sera (118 MAT confirmed positive and 97 MAT negative). The overall sensitivity was close to 85.6 % and the specificity was 83.5 %, according to the MAT reference method. Analytical specificity of the IgG-ELISA was evaluated using 50 bovine serum samples from animals showing serum antibodies against other pathogens that cause abortion in bovine, such as Brucella sp., Neospora sp. and Bovine Viral Diarrhea (BVD), and no cross-reaction was observed. This IgG-ELISA can be an alternative to the MAT for diagnosis of leptospiral infection in bovine.     

Clinical and epidemiological features of canine leptospirosis in North Queensland

OBJECTIVE: To evaluate the clinical signs, results of clinical pathology and serology tests, and treatment outcome of clinical leptospirosis in 40 dogs from North Queensland. DESIGN: Retrospective study from January 1995 to August 1999. PROCEDURE: Case records were reviewed for age, breed, sex, month of submission, geographical location and presenting clinical signs in 40 dogs with titres of > or = 200 for leptospirosis by the microscopic agglutination test. A biochemistry panel and complete blood count were performed on 18 dogs. RESULTS: Canine leptospirosis occurred most frequently during the summer and autumn particularly in the 'wet' tropical coastal areas of Mackay and Cairns. Fewer cases were seen in the Atherton Tablelands and 'dry' tropics around Townsville. Young and male dogs were more commonly affected. Most cases were caused by L australis (80%) and L zanoni (15%) with individual cases of L hardjo and L copenhageni. All dogs showed a distinctive multiorgan disease pattern including renal failure and cholestatic hepatopathy. Presenting clinical signs were related to these disease syndromes and included jaundice, vomiting, lethargy, inappetence, dehydration, pyrexia, abdominal pain and diarrhoea. Just over 50% of the dogs in the present study failed to recover. Clinical biochemical changes indicative of renal failure and cholestasis were significantly less severe in dogs that recovered compared with dogs that did not survive. CONCLUSIONS: Clinical pathology testing should be performed on all dogs presented for illness in the endemic areas so as to help make a presumptive diagnosis and assist in determining the prognosis as soon as possible. The presumptive diagnosis should be confirmed serologically.     

Investigation of a syndrome characterised by passage of red urine in smallholder dairy cattle in East Usambara Mountains, Tanzania

A case-control study was carried out to investigate a syndrome in smallholder dairy cattle in East Usambara Mountains characterised by urination of clotted blood. Smallholder dairy farms with the problem (cases) were matched with nearest farms without the problem (controls). In total, 30 farmers from Mbomole (19), Shebomeza (9) and Mlesa (2) villages in Amani division participated in the study. Using a structured questionnaire, information on risk factors associated with conditions characterised by passage of red urine in cattle was collected. In addition, serum samples from 80 smallholder dairy animals were collected and submitted for serodiagnosis of leptospirosis and babesiosis by microscopic agglutination test (MAT) and an indirect enzyme-linked immunosorbent assay (ELISA), respectively. Laboratory analysis showed that the seroprevalence of leptospirosis and babesiosis was 21.3% and 46.3%, respectively and there was no significant difference between'case' and 'control' farms (P > 0.05), hence the occurrence of urination of clotted blood syndrome in Amani was not explained. However, bracken fern (Pteridium aquilinum) was found to be ubiquitous in the area, and also found to be widespread in all areas used as sources of animal fodder Given the presence and distribution of bracken ferns and clinical signs and post-mortem lesions described by informants, chronic bracken-fern poisoning is more likely to be associated with the syndrome affecting animals in the study area. However, further investigation is required to confirm this observation so that appropriate control strategies can be devised.     

Use of a case-control study and geographic information systems to determine environmental and demographic risk factors for canine leptospirosis

Leptospirosis is increasingly diagnosed as a re-emerging canine disease in the USA. Our objectives were to describe potential risk factors for canine leptospirosis infections in northern California, through the use of a case-control study, and to perform a spatial analysis to investigate which aspects of the landscape and land use patterns are important in the transmission of leptospirosis. Forty-three cases and 59 controls were enrolled into the study. Serological results showed that 17 (39.5%) of the 43 dog cases were infected with serovar pomona. Cases were 7.86 times more likely to have been walked in a rural environment rather than an urban environment. Cases also had eight times higher odds of swimming in outdoor water, and approximately 12 times higher odds of drinking from outdoor water in the two weeks preceding illness. At smaller distances from the dogs' homes (radius or= 5 km) there was a positive relationship between leptospirosis cases and percent of wetlands or public open space. Intervention measures for the prevention of canine leptospirosis should include reducing access to potentially infectious bodies of water that are close to canine homes, and to large areas of wetlands and public open space in the general vicinity. We have shown that a spatial analysis in conjunction with traditional epidemiological analysis is a powerful combination in identifying risk factors for infectious diseases.     

Molecular Characterization by LSSP‐PCR and DNA Sequencing of a Pathogenic Isolate of Leptospira interrogans from Brazil

We report the initial characterization of a leptospiral isolate, Leptospira interrogans, serogroup Sejroe, serovar Hardjo, genotype Hardjoprajitno, strain Norma, and its relatedness with L. interrogans, serogroup Sejroe, serovar Hardjo, genotype Hardjoprajitno, strain Hardjo and Leptospira borgpetersenii, serogroup Sejroe, serovar Hardjo, genotype Hardjobovis, strain Sponselee. The Norma strain singled out during a leptospirosis outbreak in cattle immunized with antigens from the reference strain Hardjoprajitno (OMS). By applying a microscopic agglutination serological test (MAT) to cattle (n = 2966) with symptoms of leptospirosis between 2003 and 2007, more than 50% of sera were found positive for one of the following serotypes: Hardjoprajitno (31–21%), Hardjo Norma (46–40%), Hardjo hardjobovis (18–10%), Mini (8–4%) and Wolffi (7–4%). In immunization trials using six isolates plus Norma isolate, the remission of MAT in these isolates was observed following 6 months of the initial vaccination. To provide molecular ground for the high MAT Norma frequency found in these isolates, a DNA polymorphic analysis was conducted by comparing the Norma isolate with reference strains Hardjoprajitno and Sponselee. The polymorphic analysis in secY showed five base changes in Norma relative to Hardjoprajitno strain, corresponding to 98% identity, while Sponselee displayed 49 polymorphic sites relative to the Hardjoprajitno strain, representing 80% identity. The alignment of secY translated sequences shows no differences between Hardjoprajitno and Norma, and eight polymorphisms between genotype hardjoprajttno and strain Sponselee. Three‐dimensional modelling located these variations within the loop region connecting helices 7 and 8 from secY which is less conserved. DNA sequencing of 23S ribosomal conserved fragment revealed a single polymorphism between Hardjoprajitno and Norma, and 13 polymorphisms between strains Sponselee, Hardjoprajitno and Norma. The differences between Hardjo and Norma were confirmed by low stringency single‐specific primer polymerase chain reaction (LSSP‐PCR) signature experiments with the primer G2, using as template the 285 bp fragment initially amplified with G1/G2 primers.     

Sero-epidemiology of canine leptospirosis in Trinidad: serovars, implications for vaccination and public health

A sero-epidemiological study on canine leptospirosis was conducted in house, stray, farm and hunting dogs, as well as in suspect cases of clinical canine leptospirosis. Serum samples were collected from apparently healthy (vaccinated and non-vaccinated), house dogs. A questionnaire was administered to the owners to elicit information on risk factors for leptospirosis. The microscopic agglutination test was used to screen for leptospirosis using 17 international serovars. Reciprocal titres of between 100 and <800 were considered as evidence of past exposure while reciprocal titres of 800 or greater were classified as suggestive of acute/current infection. Of a total of 419 serum samples tested, 61 (14.6%) were seropositive for Leptospira agglutinins, 23 (5.5%) had mixed infections and 16 (3.8%) had current infection. Amongst 50 suspected cases of clinical leptospirosis, 24 (48.0%) were seropositive and only 13 (26.0%) had current infection compared with 10 (6.3%) and three (1.9%) of 160 apparently healthy house dogs respectively. The difference was statistically significant (P < 0.05; chi2). Twelve (25.5%) of 47 hunting dogs, 10 (20.4%) of 49 farm dogs and five (4.4%) of 113 stray dogs were seropositive (P < 0.05; chi2). Overall, a total of nine serovars were detected with serovars mankarso, icterohaemorrhagiae RGA, autumnalis and copenhageni being involved in 29 (47.5%), 20 (32.8%), 25 (41.0%) and 10 (16.4%) respectively in 61 seropositive dogs (P < 0.05; chi2). Serovar mankarso was most predominant in seropositive apparently healthy dogs, 37.8% (14/37), suspected clinical cases of leptospirosis, 62.5% (15/24) compared with serovar icterohaemorrhagiae with a frequency of 21.6% (8/37) and 50.0% (12/24), the difference being statistically significant (P < 0.05; chi2). Although all vaccines used for prevention of canine leptospirosis in the country contain serovars canicola and icterohaemorrhagiae, serovar mankarso was mostly associated with infection and disease and may be a good candidate for inclusion in the vaccine used locally. The public health risk posed to owners of dogs infected with Leptospira cannot be over-emphasized considering the zoonotic nature of the disease.     

Serovar-specific prevalence and risk factors for leptospirosis among dogs: 90 cases (1997-2002)

OBJECTIVE: To estimate serovar-specific prevalence of leptospirosis by use of veterinary teaching hospital and laboratory submission data; describe annual and seasonal patterns of leptospirosis; and identify risk factors for age, sex, and breed. DESIGN: Retrospective study. ANIMALS: 90 dogs with leptospirosis. PROCEDURES: Hospital records of dogs examined at Purdue University Veterinary Teaching Hospital with a diagnosis of leptospirosis and laboratory records of dogs from which sera were tested for antibodies against Leptospira spp at Purdue University Animal Disease Diagnostic Laboratory from 1997 through 2002 were reviewed. The likely infecting Leptospira serovar was identified. Seasonal and annual prevalences were calculated by use of hospital population at risk (hospital cases) or serologic testing submissions (diagnostic laboratory cases). Age-, sex-, and breed-specific risk factors for hospital cases were estimated by odds ratios. RESULTS: Of the 39 hospitalized dogs identified, 34 had been serologically tested, and 22 of those were infected with Leptospira kirschneri serovar grippotyphosa. Of the 51 diagnostic laboratory cases, 59% had a reciprocal titer > or = 800 against serovar grippotyphosa. Diagnostic laboratory cases were more common in summer, whereas hospital cases of leptospirosis were more common in fall. Male dogs were at significantly greater risk of leptospirosis than female dogs; and dogs 4 to 6.9 years old were at significantly greater risk than dogs < 1 year old. CONCLUSIONS AND CLINICAL RELEVANCE: L kirschneri serovar grippotyphosa infection was associated with most cases of leptospirosis in dogs. Use of an effective vaccine that includes this serovar is advisable for dogs at risk of leptospirosis.     

Evaluations of land cover risk factors for canine leptospirosis: 94 cases (2002-2009)

Associations of land cover/land use variables and the presence of dogs in urban vs. rural address locations were evaluated retrospectively as potential risk factors for canine leptospirosis in Kansas and Nebraska using Geographic Information Systems (GIS). The sample included 94 dogs positive for leptospirosis predominantly based on a positive polymerase chain reaction test for leptospires in urine, isolation of leptospires on urine culture, a single reciprocal serum titer of 12,800 or greater, or a four-fold rise in reciprocal serum titers over a 2-4 weeks period; and 185 dogs negative for leptospirosis based on a negative polymerase chain reaction test and reciprocal serum titers less than 400. Land cover features from 2001 National Land Cover Dataset and 2001 Kansas Gap Analysis Program datasets around geocoded addresses of case/control locations were extracted using 2500m buffers, and the presence of dogs' address locations within urban vs. rural areas were estimated in GIS. Multivariate logistic models were used to determine the risk of different land cover variables and address locations to dogs. Medium intensity urban areas (OR=1.805, 95% C.I.=1.396, 2.334), urban areas in general (OR=2.021, 95% C.I.=1.360, 3.003), and having urban address locations (OR=3.732, 95% C.I.=1.935, 7.196 entire study region), were significant risk factors for canine leptospirosis. Dogs regardless of age, sex and breed that live in urban areas are at higher risk of leptospirosis and vaccination should be considered.     

Experimental Leptospira interrogans Serovar Kennewicki Infection of Horses

Background: Little information is available about experimental induction of leptospirosis in horses. Objectives: Determine serologic, hematologic responses of horses to Leptospira interrogans serovar Kennewicki infection. Animals: Four adult horses seronegative for leptospirosis. Methods: Experimental and observational study. Horses were challenged with an equine isolate of L. interrogans serovar Kennewicki at 2 different doses and different inoculation sites. After challenge, the horses were monitored for 60 days. Blood, urine, and aqueous humor samples were collected at intervals until euthanasia 60 days after infection. Results: Pyrexia (39.3–40°C) occurred as early as 1 day after challenge with 10 × 10⁸Leptospira divided equally between topical ocular and intraperitoneal injection in 2 horses. Leptospires were recovered from the blood and urine but not from the aqueous humor of the 2 febrile horses. The sera of all 4 challenged horses developed microscopic agglutination test antibody after challenge and remained relatively constant for 21 days. Titer to cross‐reacting strains declined earlier than titer to the challenge strain. Conclusions: Clinical disease in experimentally infected horses can be mild or inapparent in Leptospira infected horses. Repeated serologic testing can allow recognition of the infecting serovar. In febrile horses, Leptospira can be isolated from blood while isolation from the urine can occur after fever has subsided.