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Eimeria bovis is a pathogenic protozoan that causes cattle digestive tract infections, which can cause economic losses to farmers. It is necessary to develop specific and accurate detection methods to conserve livestock and prevent coccidiosis in Indonesia. This study aims to detect E. bovis by nested PCR and determine the relationship with reference sequences. A total of 167 samples of beef cattle feces were taken randomly from community farms spread across 18 provinces in Indonesia. The feces were examined natively, and then the oocysts were purified by the sugar flotation method, extracted by KIT extraction, and amplified by the nPCR technique. Positive samples were followed by sequencing and phylogenetic analysis using MEGA 11 software. This study used two pairs of primers (outer and inner) taken from ITS-1 molecular markers. As many as 96 out of 167 samples (57.5%) were positive for Eimeria spp., and 48 of the 96 samples were positive for Eimeria spp. (50%) were detected to be positive for E. bovis based on the presence of a 238 bp DNA fragment. The results of the phylogenetic analysis showed that the study sample formed a separate cluster from the E. bovis cluster from abroad. In conclusion, E. bovis was detected in 16 out of 18 provinces in this study, and the nPCR technique proved to have better sensitivity and specificity. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii pathogenicity depends on the type derived from a clonal population. A genetic analysis of the locus has been carried out to determine the different genotypes of T. gondii (strain types I, II, and III) that are associated with human toxoplasmosis. The several genotypes of T. gondii (strain types I, II, and III) that are linked to human toxoplasmosis have been identified through genetic study of the locus. In this investigation, PCR-RFLP was found to be a useful, and simple method genotypic characterization. The objective of this study was to genotyping characterize T. gondii RH and BEV strains isolates by PCRRFLP using several restriction enzymes. T. gondii tachyzoite DNA was extracted and amplified by PCR using dense granule genetic markers (GRA1 and GRA7) designed with Primer3plus. The amplification were digested using the restriction enzymes. The PCR-RFLP amplified dense granule products was used to classify strains into two genotypes of T. gondii (virulent and avirulent). The results demonstrated that the RFLP patterns of the GRA1 and GRA7 gene area digested by DdeI, MvaI, HinfI, RsaI, and Sau96I enzymes can be used to identify virulent or avirulent strains of T. gondii. Toxoplasma gondii RH and BEV strain produced different digestion product which can be used to distinguished the strains. [ABSTRACT FROM AUTHOR]

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