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. 2023 Oct-Dec;18(4):554-562.
doi: 10.18502/ijpa.v18i4.14264.

Molecular Characterization of Liver Fluke Isolated from Sheep, Goat and Cattle in Sulaymaniyah, Iraq

Vilya Shwan Othman  1 , Abdullah Ahmed Hama  1   2 , Rostam Hama Zorab  3 , Abdolhossein Dalimi  4
Affiliations

Molecular Characterization of Liver Fluke Isolated from Sheep, Goat and Cattle in Sulaymaniyah, Iraq

Vilya Shwan Othman et al. Iran J Parasitol. 2023 Oct-Dec.

Abstract

Background: We aimed to determine species of liver fluke that predominately cause fascioliasis in sheep, goats, and cattle in the Sulaymaniyah Province, Iraq using the molecular technique of DNA sequencing and restriction fragment length polymorphism (RFLP).

Methods: The samples were collected from November 2021 to May 2022. The flukes were collected from infected livers of livestock at the slaughterhouse of Sulaymaniyah Governorate, Iraq. A total of 205 flukes were collected from 56 hosts, cattle (n=22), sheep (n=28), and goats (n=6). The specific primers for FCOX1 and 28S rDNA gene amplification were used. The PCR products were subjected to restriction fragment polymorphism (RFLP) assay using Hpy188III and Dra II restriction enzymes, besides DNA sequencing.

Results: The results showed the genetic polymorphisms among the flukes. Three patterns of RFLP were observed Fasciola hepatica, F. gigantica, and F. intermediate, where 28 of them displayed F. hepatica (sheep, n=14, goat, n=3 and cattle, n= 11), whereas 24 samples displayed the F. gigantica (sheep, n=12, goat, n=3 and cattle, n= 9), and only four samples belonged to F. intermediate (sheep n=3 and cattle, n=1). In addition, the result of the ribosomal DNA (28S rDNA) sequencing confirmed that the isolated flukes belonged to F. hepatica, F. gigantica and F. intermediate.

Conclusion: All three main species are present in the study area and F. hepatica predominated among the animal species in this area also, our results concluded that PCR-RFLP is a rapid and reliable method for liver fluke species identification.

Keywords: Fasciola gigantica; Fasciola hepatica; Iraq; Ruminants.

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Conflict of interest statement

Conflict of interests The authors of this study confirmed that we have no kind of conflict of interest.

Figures

Fig. 1:
Fig. 1:
(A) Gel electrophoresis of PCR products of ribosomal DNA primer. M= DNA ladder, 1–4 samples isolated from cattle, 5–9 samples isolated from sheep, 10–12 samples isolated from goat, N= negative control. (B) Gel electrophoresis of PCR products of FCOX1 primer. M= DNA ladder, 1–3 samples isolated from cattle, 4–7 samples isolated from sheep, 8–9 samples isolated from goat, N= negative control
Fig. 2:
Fig. 2:
Restriction fragment length polymorphism (RFLP) Patterns of PCR products of FCOX1 primer digested with Hpy188III enzyme: M= DNA Ladder, 1–9= the samples isolated from sheep, 10–15= samples isolated from cattle and 16–20 = samples isolated from goat
Fig. 3:
Fig. 3:
Restriction fragment length polymorphism (RFLP) Patterns of PCR products of 28s rDNA primer digested with DraII enzyme: M= DNA Ladder, 1–7= the samples isolated from sheep, 8–15=the samples isolated from cattle, 16–20= the samples isolated from goat
Fig. 4:
Fig. 4:
The partial sequence alignment of the PCR product of 28S rDNA show the genetic variation among F. hepatica and intermediate form (sq6,7,8, 9 and 12= F. hepatica and sq10 = intermediate form)
Fig. 5:
Fig. 5:
The partial sequence alignment of the PCR product of 28S rDNA show the genetic variation among F. gigantica (sq1,2,3, 4,5 and 11= the samples belong to F. gigantica)
See this image and copyright information in PMC

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