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Abstract

The study established the morphologic and molecular differentiation of Fasciola hepatica and Fasciola gigantica in buffaloes. Specifically, the study described the gross structure and morphometry of F. hepatica and F. gigantica and validated the identification of Fasciola spp. based on gross morphology and PCR results. Sixty (60) samples were evaluated grossly and morphometrically using body length, body width, cone width, and cone length as parameters to differentiate the two species. Ten representative samples from each species identified based on the parameters were subjected to single step-duplex polymerase chain reaction (PCR) for molecular identification of the species identity. Results showed significant differences in body length, cone width and cone length between F. gigantica-like and F. hepatica-like samples. F. hepatica-like samples have significantly shorter (P=0.001) body length (29.20±3.04mm) as compared to F. gigantica-like samples (46.23±4.54mm). Mean cone width was also found to be significantly (P=0.001) longer in F. gigantica-like (3.67±0.55mm) compared to F. hepatica-like (3.33±0.48mm Cone length was significantly higher (P=0.001) in F. gigantica (3.67±0.55mm) than F. hepatica (2.90±0.33mm). Body width (BW) showed no significant difference (P=0.186) between the two species. F. gigantica-like samples had mean BW measurements of 10.63±0.99mm which had a very slight difference with F. hepatica-like samples at 10.33±0.71mm. PCR products obtained from the single-step duplex PCR targeting either the cytochrome c oxidase subunit I or the large subunit ribosomal RNA revealed that all 20 samples that were morphometrically differentiated as F. gigantica or F. hepatica were indicative of F. gigantica identity. The present findings suggested that conventional gross and morphometric analysis of Fasciola spp. alone was not completely reliable in differentiating the two species thus a use of molecular identification assay would yield a more dependable result.