Detection of Pathogenic Leptospira spp. in Swamp Buffalo Bulls Frozen Semen by Polymerase Chain Reaction Technique

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Inthiporn Pheangrat
Prayout Sarkaou
Thuchadaporn Chaikhun-Marcou

Abstract

Leptospirosis is an important zoonotic disease, especially for farmers and those involved in animal husbandry. Infected swamp buffaloes usually are asymptomatic but infertility issues may present themselves. This study was designed to detect pathogenic Leptospira spp. by polymerase chain reaction (PCR) technique in commercially available and widely used frozen semen samples from 56 swamp buffalo bulls in Thailand. DNA samples were extracted and grouped into seven experimental group samples (eight samples per test tube). Polymerase Chain Reaction assays were performed using single primer LipL32 and 16S rRNA and multiplex primer Ph In and Sa. The results showed that Leptospira spp. was not detected in any of the seven experimental group samples. However, penicillin and streptomycin were added in the extenders of these frozen semen samples, which could have acted as antibiotics for Leptospira spp. or The buffalo bulls were healthy without Leptospira spp. Infection.

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Research Article

References

Andrabi, S. M., Khan, L. A. and Shahab, M. 2016. Isolation of bacteria in semen and evaluation of antibiotics in extender for cryopreservation of buffalo (Bubalus bubalis) bull spermatozoa. Andrologia. 1-9.

Ansari, M., Rakha, B. and Akhter, S. 2017. Cryopreservation of Nili-Ravi buffalo (Bubalus bubalis) semen in AndroMed® extender; in vitro and in vivo evaluation. Reproduction in Domestic Animals. 2017: 1-6.

Centers for Disease Control and Prevention (CDC). 2019. Leptospirosis: Systematic review of the published literature. https://www.cdc.gov/leptospirosis/index.html. (Accessed 25 August 2022).

Chadsuthi, S., Bicout, D. J., Wiratsudakul, A., Suwancharoen, D., Petkanchanapong, W., Modchang, C., Triampo, w., Ratanakorn, p. and Chalvet-Monfray, K. 2017. Investigation on predominant Leptospira serovars and its distribution in humans and livestock in Thailand, 2010-2015. PLOS Neglected Tropical Diseases. 10.1371: 1-18.

Chadsuthi, S., Chalvet-Monfray, K., Wiratsudakul, A., Suwancharoen, D. and Cappelle, J. 2018. A remotely sensed flooding indicator associated with cattle and buffalo leptospirosis cases in Thailand 2011–2013. BMC Infectious Diseases. 18(602): 1-9.

Chaikhun, T., Tharasanit , T., Rattanatep, J.,De Rensis, F. and Techakumphu, M. 2010. Fertility of swamp buffalo after synchronization of ovulation by sequential GnRH and PGF administration (ovsynch) and fixed-timed artificial insemination. Theriogenology. 74: 1371-1376.

Department of Disease Control (DDC). 2021. Leptospirosis. Systematic review of the published literature. https://ddc.moph.go.th/brc/news.php?news=26719&deptcode=b rc. (Accessed 25 August 2022).

Department of Livestock Development (DLD). 2022. Farmer/Livestock Information, Fiscal Year 2022. Systematic review of the published literature. http://ict.dld.go.th/webnew/ index.php/th/service-ict/report/396-report-thailand-livestock/reportservey2565. (Accessed 25 August 2022).

Fraga, T. R., Carvalho, E., Isaac, L. and Barbosa, A. S. 2015. Leptospira and Leptospirosis. Animal and Ectoparasitic Source Infections. 1973-1990.

Ganda, M. R., Souza, M. A., Lourencetti, M. P., Ciuffa, A. Z., Soares, P. M. and Lima, A. M. 2016. Molecular detection of Leptospira spp. in bull semen. Revue de Médecine Vétérinaire. 167(11-12): 297-301.

Givens, M. D. 2018. Review: Risks of disease transmission through semen in cattle. Animal. 12(S1): 165-171.

Glorian, N. G., Villanue, S. Y. A. M., Yanagiha, Y. and Yoshid, S. 2016. Identification of Prevalent Leptospira Serovars Infecting Water Buffaloes, Cows, Dogs, Pigs and Rats in the Philippines. Southeast Asian J Trop Med Public Health. 47(4): 766-773.

Guedes, I. B., Souza, G. O., Paula Castro, J. F., Souza Filho, A. F., Cavalini, M. B., Taniwaki, S. A., Maia, A. L. P., Pereira, I. C. and Heinemann, M. B. (2020). Identification of Pathogenic Leptospira Species in the Urogenital Tract of Water Buffaloes (Bubalus bubalis) From the Amazon River Delta Region, Brazil. Frontiers in Veterinary Science, 7(269), 1-7.

Hamond, C., Martins, G., Medeiros, M. A. and Lilenbaum, W. 2013. Presence of Leptospiral DNA in Semen Suggests Venereal Transmission in Horses. Journal of Equine Veterinary Science. 33:1157-1159.

Heinemann, M. B., Garcia, J. F., Nunes, C. M., Gregori, F., Higa, Z. M., Vasconcellos, S. A. and Richtzenhain, L. J. 2000. Detection and differentiation of Leptospira spp. serovars in bovine semen by polymerase chain reaction and restriction fragment length polymorphism. Veterinary Microbiology, 73, 261-267.

Lilenbaum, W., Varges, R., Brandao, F. Z., Cortez, A., de Souza, S. O., Brandao, P. E., Richtzenhain, L. J. and Vasconcellos, S. A. 2008. Detection of Leptospira spp. in semen and vaginal fluids of goats and sheep by polymerase chain reaction. Theriogenology. 69:837-842.

Loureiro, A. P. and Lilenbaum, W. 2020. Genital bovine leptospirosis: A new look for an old disease. Theriogenology. 141:41-47.

Maiolino, S. R., Cortez, A., Langoni, H., Giuffrida, R., Santos, J. R., Júnior, G. D., Lara, G. H. B., Motta, R. G., Chacur, M. G. M., Monteiro, F. M., Heinemann, M. B., Filho, A. F. S., Martins, L. S. A., Bello, T. S. and Ribeiro, M. G. 2021. Sperm viability, serological, molecular, and modified seminal plasma agglutination tests in the diagnosis of Leptospira in the semen and serum of bovine bulls. Brazilian Journal of Microbiology. 1-8.

Masri, S. A., Nguyen, P. T., Gale, S. P., Howard, C. J. and Jung, S. C. 1997. A Polymerase Chain Reaction Assay for the Detection of Leptospira spp. in Bovine Semen. Canadian Journal of Veterinary Research, 61, 15-20.

Mughal, D. H., Ijaz, A., Yousaf, M. S., Wadood, F. and Farooq, U. 2017. Cryopreservation of buffalo (Bubalus bubalis) semen-limitations and expectations. Buffalo Bulletin. 36(1): 1-14.

Na-Chiangmai, A. 2002. Raising buffaloes. Department of Livestock Development. Bangkok. p. 1-9.

Perumal, P., K., C. J., Srivastava, N., Vupru, K. and Srivastava, S. K. 2013. Infectious Causes of Infertility in Buffalo Bull (Bubalus bubalis). International Journal of Bio-resource and Stress Management. 4(1):084-090.

Philip, N., Garba, B. and Neela, V. K. 2018. Long-term preservation of Leptospira spp.: challenges and prospects. Applied Microbiology and Biotechnology. 102: 5427–5435.

Philpott, M. 1993. The dangers of disease transmission by artificial insemination and embryo transfer. British Veterinary Journal. 149(4): 339-369.

Pui, C. F., Bilung, L. M., Su'ut, L. and Apun, K. 2015. Prevalence of Leptospira Species in Environmental Soil and Water from National Parks in Sarawak, Malaysia. Borneo Journal of Resource Science and Technology, 5(1), 49-57.

Suwancharoen, D., Chaisakdanugull, Y., Thanapongtharm, W. and Yoshida, S. 2013. Serological survey of leptospirosis in livestock in Thailand. Epidemiology and Infection. 141:2269-2277.

Suwancharoen, D., Limlertvatee, S., Chetiyawan, P., Tongpan, P., Sangkaew, N., Sawaddee, Y., Inthakan, K. and Wiratsudakul, A. 2016. A nationwide survey of pathogenic leptospires in urine of cattle and buffaloes by Loop-mediated isothermal amplification (LAMP) method in Thailand, 2011–2013. The Journal of Veterinary Medical Science. 78(9):1495-1500.

Uavechanichkul, R. 2008. Guidelines for Quality Control of Semen Quality for Artificial Insemination. Department of Livestock Development. Bangkok. p. 36-116.

Vital-Brazil, J. M., Balassiano, I. T., Oliveira, F. S., Souza Costa, A., Hillen, L. and Pereira, M. M. 2010. Multiplex PCR-based detection of leptospira in environmental water samples obtained from a slum settlement. Memórias do Instituto Oswaldo Cruz, 105(3), 353-355.

Wongpanit, K., Suwanacharoen, D. and Srikram, A. 2012. Serological Survey of Leptospirosis in Thai Swamp Buffalo (Bubalus bubalis) in Sakon Nakhon Province, Thailand. Kasetsart Journal (Natural Science). 46:736-741.