Detection and species identification of Cryptosporidium by using immunofluorescent assay (IFA) and PCR sequencing from biogas waste in Bac Giang

Abstract

This study was done to access the difference and effectiveness of using both IFA and PCR sequencing methods in species identification and confirmation of presence of Cryptosporidium spp. in biogas waste. In this study, 167 biogas waste samples were collected from biogas influent and effluent samples in three communes (Viet Yen, Lang Giang, and Hiep Hoa) in Bac Giang province frp, 2019 to 2020 to test for Cryptosporidium spp., and Giardia spp., (oo)cysts. Among these samples, 16 samples analyzed by immunofluorescent assay showed the presence of Cryptosporidium spp.,. Of which 11 samples were simultaneously confirmed by polymerase chain reaction. Nucleic acid sequencing was done on the 18S rRNA gene. Four Cryptosporidium scrofarum, and 2 C. suis were identified in biogas influent samples. Two C. scrofarum, 1 C. suis, 1 C. bailey, and 1 C. meleagridis were detected in biogas effluent samples. The sequencing results showed that the isolated Cryptosporidium spp. are zoonotic species. Therefore, the identification of Cryptosporidium spp. by IFA and PCR sequencing has important implications in epidemiological studies, investigation of infection/contamination origin, or surveillance of diarrhea caused by Cryptosporidium.