Colorimetric detection of Tritrichomonas foetus by a LAMP assay in preputial fluid and cryopreserved semen from bulls raised in Sonora, Mexico
DOI:
https://doi.org/10.17533/udea.rccp.e361359Keywords:
biosafety, esmegma, LAMP, molecular diagnostics, semen, trichomoniasisAbstract
Background: Bovine trichomoniasis, caused by Tritrichomonas foetus, is a venereal disease that affects the reproductive efficiency of cattle, especially in extensive breeding systems. Infected bulls could spread the protozoan by artificial insemination if the semen is contaminated. The loop-mediated isothermal amplification (LAMP) is an alternative assay to detect genetic material of pathogens without the need to purify inhibitors contained in a biological sample. Objective: Detection of T. foetus in cryopreserved semen and genital fluids from bulls raised in Sonora Mexico by a colorimetric LAMP assay. Methods: The conditions for a colorimetric LAMP assay were standardized using PCR as analytical reference in order to detect T. foetus (crLAMP-TF). A total of 110 preputial samples were tested by crLAMP-TF. From positive males to T. foetus (TF+), seminal samples were collected, cryopreserved and tested by crLAMP-TF. Results: The crLAMP-TF assay was sensitive, and allow the visual detection at 35 min. without inhibitors purification of biological samples previous to DNA extraction. The 29.1% of the bulls were bTF+ and 27.3% of the thaws were contaminated. Conclusion: This survey confirms the presence of T. foetus in Sonora, Mexico and the ability of the protozoan to contaminate semen, which represents a potential risk of transmission during assisted reproduction programs by artificial insemination. The crLAMP-TF assay represents a viable alternative against traditional molecular techniques and its potential in the epidemiological surveillance of pathogens under field conditions.
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