Background Leptospirosis, commonly known as rat-urine disease, is a global but endemic zoonotic disease in the tropics. Despite the historical report of leptospirosis in Malaysia, the information on human-infecting species is limited. Determining the circulating species is important to understand its epidemiology, thereby to strategize appropriate control measures through public health interventions, diagnostics, therapeutics and vaccine development. Methodology/Principle findings We investigated the human-infecting Leptospira species in blood and serum samples collected from clinically suspected leptospirosis patients admitted to three tertiary care hospitals in Malaysia. From a total of 165 patients, 92 (56%) were confirmed cases of leptospirosis through Microscopic Agglutination Test (MAT) (n = 43; 47%), Polymerase Chain Reaction (PCR) (n = 63; 68%) or both MAT and PCR (n = 14; 15%). The infecting Leptospira spp., determined by partial 16S rDNA (rrs) gene sequencing revealed two pathogenic species namely Leptospira interrogans (n = 44, 70%) and Leptospira kirschneri (n = 17, 27%) and one intermediate species Leptospira wolffii (n = 2, 3%). Multilocus sequence typing (MLST) identified an isolate of L. interrogans as a novel sequence type (ST 265),

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Introduction. Municipal service workers have been found to have an occupational risk of leptospirosis. Study among municipality workers shows high seropositivity of leptospiral antibodies detected among town cleaners and garbage collectors.Objective. Aims of this study were to determine seroprevalence of leptospiral antibodies and distribution of serovars detected in samples among municipal service workers.Methodology. Cross-sectional study involved 89 municipal service workers in Selangor. Blood samples were taken and serological test was done using MAT following standard procedures.Results. Seropositivity of leptospiral antibodies among municipal service workers was 34.8%. Serovars identified were strains ofSarawak, Copenhageni, Hardjobovis, Lai, Bataviae, Patoc, Celledoni, Hardjoprajitno, Tarrasovi, andPomona. There were 31 workers with positive leptospiral antibodies. All of them were frequently exposed towards leptospirosis. Significant associations have been reported between seropositivity of leptospiral antibodies with job category (P=0.021) and worker’s nationality (P=0.014) among municipal service workers.Conclusion. High seropositivity of leptospiral antibodies detected among municipal service workers which was associated with job category and nationality of workers. The significant findings from this study suggest that health education programs and safe work practice should be considered to prevent leptospirosis among municipal service workers in future.

Little is known on the knowledge, attitudes and preventive practices (KAP) of leptospirosis worldwide. This study embarked on assessing the KAP of leptospirosis among rural communities in Malaysia. A total of 444 participants (223 male; 221 female) aged between 18 and 81 years old were surveyed by using a self-administered questionnaire. A majority of participants had poor knowledge level (57.0%), unacceptable attitudes (90.3%) and unacceptable preventive practices (69.1%) on leptospirosis, and only 29.7% knew “rat-urine disease” as leptospirosis. Only 34.2% of the participants knew the bacteria could enter via wound lesions. Ethnicity and income were strongly associated with knowledge level and preventive practices, respectively (p-values < 0.05). As for attitudes, ethnicity, income and education type were significantly associated (p-values < 0.05). Only 36.5% of the participants were willing to see a doctor and did not mind if their house or surrounding area is dirty (59.7%). Surprisingly, only 32.9% had used rubber boots during floods. By logistic regression analysis, ethnicity was the only significant predictor for both knowledge level (an odds ratio (AOR) = 0.39, 95% confidence interval (CI) = 0.222–0.680) and preventive practices (AOR = 1.81, 95% CI = 1.204–2.734). Ethnicity (AOR = 0.40, 95% CI = 0.239–0.665), income (AOR = 1.58, 95% CI = 1.041–2.385) and education type (AOR = 3.69, 95% CI = 1.237–10.986) were strong predictors for attitudes. Among the KAP variables, attitude (AOR = 4.357, 95% CI = 2.613–7.264) was the only predictor for the preventive practices by logistic regression analysis. The KAP elements on leptospirosis are still lacking and poor health seeking behavior and attitudes are of our utmost concern. Thus, effective strategies should be planned to impart knowledge, and develop proactive approaches and good preventive modules on leptospirosis to this leptospirosis-prone community.

The diagnosis of leptospirosis remains a challenge due to its non-specific symptoms and the biphasic nature of the illness. A comprehensive diagnosis that includes both molecular (polymerase chain reaction (PCR)) and serology is vital for early detection of leptospirosis and to avoid misdiagnosis. However, not all samples could be subjected to both tests (serology and molecular) due to budget limitation, infrastructure, and technical expertise at least in resource-limited countries. We evaluated the usefulness of testing the clinically suspected leptospirosis cases with both techniques on all samples collected from the patients on the day of admission. Among the 165 patient's blood/serum samples tested (from three hospitals in Central Malaysia), 43 (26%) showed positivity by microscopic agglutination test (MAT), 63 (38%) by PCR, while 14 (8%) were positive by both MAT and PCR. For PCR, we tested two molecular targets (lipL32 by qPCR and 16S rDNA or rrs by nested PCR) and detected lipL32 in 47 (29%) and rrs gene in 63 (38%) patients. The use of more than one target gene for PCR increased the detection rates. Hence, a highly sensitive multiplex PCR targeting more than one diagnostic marker is recommended for the early detection of Leptospira in suspected patients. When the frequencies for positivity detected either by MAT or PCR combined, leptospirosis was diagnosed in a total of 92 (56%) patients, a higher frequency compared to when samples were only tested by a single method (MAT or PCR). The results from this study suggest the inclusion of both serology and molecular methods for every first sample irrespective of the days post-onset of symptoms (DPO) collected from patients for early diagnosis of leptospirosis.