Publication: Optimization of culture protocols to isolate Leptospira spp. From environmental water, field investigation, and identification of factors associated with the presence of Leptospira spp. From the environment
Issued Date
2020-06-01
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ISSN
24146366
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2-s2.0-85087499931
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Mahidol University
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SCOPUS
Bibliographic Citation
Tropical Medicine and Infectious Disease. Vol.5, No.2 (2020)
Suggested Citation
Udomsak Narkkul, Janjira Thaipadungpanit, Janjira Thaipadungpanit, Prapaporn Srilohasin, Preeraya Singkhaimuk, Metawee Thongdee, Somjit Chaiwattanarungruengpaisan, Panadda Krairojananan, Wirichada Pan-Ngum Optimization of culture protocols to isolate Leptospira spp. From environmental water, field investigation, and identification of factors associated with the presence of Leptospira spp. From the environment. Tropical Medicine and Infectious Disease. Vol.5, No.2 (2020). doi:10.3390/tropicalmed5020094 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/57971
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Title
Optimization of culture protocols to isolate Leptospira spp. From environmental water, field investigation, and identification of factors associated with the presence of Leptospira spp. From the environment
Abstract
© 2020 by the authors. The successful culture of Leptospira spp. from the environment is challenging. Here, we optimized the isolation of Leptospira spp. from water samples spiked with different species and initial concentrations of this organism. The time periods between water sampling and the isolation process were varied (0, 2, and 4 weeks). Bacterial cultures were observed under a microscope, and cultures were graded for cell density, weekly, for 12 weeks. Most pathogenic Leptospira spp. were difficult to culture under all conditions. All conditions of water samples spiked with novel species of Leptospira subclade P1 were culture positive within 2 weeks. For Leptospira subclade P2, storing samples for 2 weeks prior to isolation resulted in more successful isolation compared with isolation after other storage conditions. For subclade S1, all samples with initial bacterial concentrations of more than 103 colonies/mL, under all storage conditions, were successfully cultured. These results suggest that storing contaminated water samples for 2 to 4 weeks in the dark at an ambient temperature prior to culturing can improve the isolation of Leptospira spp. from the samples. We implemented this protocol and collected water samples from natural sources accessed by both humans and animals. Leptospira spp. was identified in 32% (35/109) of water samples. The animal species using a water source influenced the likelihood of water samples being contaminated with Leptospira spp. Cultures of Leptospira spp. from environmental samples can provide useful information for understanding the complex interactions between humans, animals and the environment in the transmission of leptospirosis.