Cloning and expression of Fasciola hepatica enolase gene and efficacy of recombinant protein in the serodiagnosis of sheep fasciolosis.

Fasciolosis caused by Fasciola hepatica is a disease of zoonotic importance that is common worldwide and can cause serious problems in farm animals, some wild animals and humans. The development of diagnostic kits for the correct diagnosis of fasciolosis in sheep is important in terms of preventing yield losses. With this study, it is aimed to clone and express the enolase gene to be isolated from adult F. hepatica and to determine the effectiveness of the recombinant antigen in the serodiagnosis of sheep fasciolosis. For this aim, primers were designed to amplify the enolase gene from the F. hepatica enolase sequence, mRNA was isolated from F. hepatica adult fluke obtained from an infected sheep followed by cDNA was obtained. Enolase gene was amplified by PCR and the product was cloned and then expressed. The efficiency of the purified recombinant protein was displayed by Western blot (WB) and ELISA using positive and negative sheep sera. As a result, the sensitivity and specificity rates of the recombinant FhENO antigen were 85% and %82.8 by WB while the rates were 90% and 97.14% by ELISA, respectively. At the same time, in sheep blood sera samples collected from the Elazig and Siirt provinces of Turkey, 100 (50%) of 200 sera were found to be positive by WB and 46 (23%) were found to be positive by ELISA. The most important problem in ELISA was the high cross-reaction rate of the recombinant antigen used, as in WB. In order to prevent the cross-reactions, it will be useful to compare the genes encoding the enolase protein of parasites from the closely related parasite family, and select the regions where there are no common epitopes, and clone them and test the purified protein.

Investigation of the Prevalence of Fasciola hepatica in Small Ruminants in the Siirt Region, Turkey.

BACKGROUND: Fasciolosis is a disease of the liver caused by trematodes in the family of Fasciolidae, particularly by Fasciola hepatica and Fasciola gigantica. The aim of this study was to investigate the prevalence of F. hepatica in sheep using the ELISA method, and in hair goats by post-mortem liver examination in the Siirt region, Turkey. METHODS: This study was conducted between Feb-Sep 2018. Five ml of blood samples were taken from the jugular veins of 320 sheep, which were selected from various locations of Siirt region by random sampling method. Fasciolosis seroprevalence in sheep was investigated by the ELISA method, using commercial kits (BIOK 211-Monoscreen AB ELISA F. hepatica test). In order to determine the prevalence of fasciolosis in hair goats, 580 slaughtered goats were examined for F. hepatica by incisions in the liver, gallbladder, and bile ducts. RESULTS: While 24 (7.50%) sheep were seropositive, 296 (92.50%) were seronegative. Regarding the hair goats, on the other hand, 82 (14.14%) were positive, while 498 (85.86%) were negative. CONCLUSION: F. hepatica infection causes significant economic losses due to the destruction of the liver in small ruminants. Considering zoonotic properties of the disease, it has been concluded that the necessary measures should be taken and anti-helminthic drugs should be applied to the animals that come out of the pasture. Furthermore, periodic examinations should be conducted, and the breeders should be informed about the disease to raise awareness.