Detection of Cryptosporidium parvum DNA in fecal samples of infected cattle (Bos indicus) and water buffaloes (Bubalus bubalis) in the Philippines using loop mediated isothermal amplification method.

Abstract

Fecal DNA samples from 17 cattle and 38 water buffaloes found to be infected with Cryptosporidium oocysts using Kinyoun acid fast stain from a previous study, were subjected to loop-mediated isothermal amplification (LAMP) assay using specific primers for Cryptosporidium parvum (C. parvum) from three municipalities, Maria, Baler and San Luis of the province of Aurora in the Philippines. Results of the fecalysis using Kinyoun acid fast stain and LAMP assay were compared with the PCR results of the examined farmer/owner who raised these animals to determine the possible zoonoses of C. parvum between the farmers and their animals. Using LAMP assay, only 41% (7/17) were positive in cattle and 76% (29/38) in water buffaloes. Out of the seven LAMP positive cases in cattle, 86% (6/7) came from Maria and 14% (1/7) from Baler. Out of 29 LAMP positive cases in water buffaloes, 62% (18/29) came from Maria, 24% (7/29) from Baler and 14% (4/29) from San Luis. Comparing with the earlier results for probable zoonoses of C. parvum between the farmers and their animal was determined. Eight farmers that were positive in PCR and with their water buffaloes, positive in LAMP assay were detected to have C. parvum. Only one farmer with his cattle was detected positive of Cryptosporidium spp. in PCR, however, it was negative in LAMP assay hence, a non-parvum species might infected the farmer and the animal.