DNA extraction methods for molecular detection of Eimeria spp. in cattle and sheep

Abstract

Molecular detection of Eimeria species in fecal samples can be useful for experimental and diagnostic purposes. However, the parasite quantity presence in feces and the oocyst wall are an obstacle in DNA extraction protocols. Therefore, adequate sampling and effective disruption of the oocysts are essential to improve the accuracy of DNA detection by PCR. The aims of this study were to evaluate the suitability of six protocols for DNA extraction from Eimeria spp. present in bovine and sheep. Twenty pools of fecal samples from cattle (10 pools) and sheep (10 pools) were distributed to six DNA extraction protocols: commercial kit, commercial kit with modification, DNAzol, cetyl-trimethyl ammonium bromide (CTAB), glass beads and commercial kit for fecal samples. Fecal samples were submitted to DNA extraction and PCR. Among the protocols tested, CTAB was determined to be most suitable for DNA extraction from oocysts (90% of DNA detection by PCR); DNAzol and CTAB resulted in higher DNA detection from bovine samples (80%). CTAB and commercial kit with modification improved PCR detection of Eimeria spp. in sheep samples, with positive amplification of DNA in all tested samples.

Coccidiosis in Small Ruminant and Antiparasitic Activity of Essential Oils

“…Oocysts per gram of feces (OPG) were examined using the modified McMaster technique with saturated saline solution to determine the parasitic load (Hao et al, 2018). Molecular detection of Eimeria species in fecal samples helpful for experimental and diagnostic purposes (Reginato et al, 2020). Morphology of sporulated Eimeria oocysts in feces of black goats in Liangshan, Chinashown in figure 2.…”