Development of Cytochrome B, a new candidate gene for a high accuracy detection of Fasciola eggs in fecal specimens

Abstract

Fasciolosis among domestic ruminants has resulted in a decrease in the production of milk products and has occasionally led to the deaths of young ruminants due to of acute infections. This study aimed to discriminate between the eggs of Fasciola gigantica and other trematode eggs in samples collected from ruminant feces specimens using PCR-based methods with the new candidate gene Cytochrome B (CYTB). A species-specific primer was developed with a high degree of sensitivity (3.285 pg). The primer was able to amplify the F. gigantica genomic DNA and there were no positive results with the other related trematodes (Paramphistomum sp., Orthocoelium sp., Fischoederius sp., Calicophoron sp., Echinostoma revolutum, E. cinetorchis, E. ilocanum and Isthmiophora hortensis), freshwater snails (Lymnaea auricularia, Bithynia siamensis, Indoplanorbis exustus, Melanoides tuberculata, Tarebia granifera) or definitive hosts (Bos primigenius and Bubalus bubalis). The minimum concentration of DNA from eggs that could be give a positive result was 3.285 pg. Moreover, the results of the study confirmed the existence of F. gigantica in Nakhon Pathom Province with a high prevalence (28.57%) and revealed the area of infection through epidemiological mapping. Thus, the species-specific primer and epidemiological data in this study may be helpful for use in epidemiological studies, phylogenetic studies and veterinary studies in the future. © 2019 Elsevier B.V.