Molecular Characterization of Animal Fasciola Spp. Isolates from Lorestan Province, Western Iran

Abstract

Background: We aimed to detect the genetic diversity of samples identified morphologically as Fasciola spp. from sheep, cattle and goat from Lorestan Province, western Iran using PCR-RFLP method. Besides, we evaluated the genetic diversity indices, sequencing and phylogenetic analysis using mitochondrial gene (ND1 and CO1).
 Methods: PCR-RFLP analysis of ribosomal ITS1 fragment by RsaI restriction enzyme to investigate the genetic characteristics of Fasciola species obtained from different hosts (18 sheep, 21 cattle, and 17goats) was conducted. The samples were sequenced. Sequences were evaluated using BLAST software and the parasite species were identified with similarity percentage and overlap with the species registered in the gene bank. Then similarity and diversity of intra-species and intra-species diversity of Fasciola species were calculated.
 Results: In Lorestan, based on RFLP pattern, 93% (52) of the Fasciola spp. isolates had a RFLP pattern related to F. hepatica and 7% (4) were F. gigantica. No hybrid forms were detected. The CO1 gene could clarify 19 haplotypes against ND1 gene that found 22 haplotypes among livestock. Sequencing results of the mtDNA showed intra-species identity 98. 5%-100% and Intra-species-diversity: 0-1.5% compared to the GenBank sequences.
 Conclusion: Using PCR-RFLP method, two species of F. hepatica and F. gigantica, were present in Lorestan Province, but F. hepatica was more prevalent. Mitochondrial genes could better test variability indices in different hosts than ribosomal genes, consequently among mitochondrial genes, the ND1 gene could better examine differences and similarities than CO1.

Seroprevalence of Fasciola sp. and Toxoplasma gondii Infections in Rural and Urban Inhabitants of Jolfa County, Northwest Iran

“…The highest prevalence rate of fascioliasis in animals is reported in the north, and the lowest prevalence is in Central Iran [20]. Heydarian et al [21] investigated the seroprevalence of human fascioliasis in Lorestan Province, Western Iran, on 1256 humans. They detected anti-Fasciola antibodies in 16 individuals (1.3%).…”

Molecular characterization of Fasciola spp. from ruminants in Duhok province using the ITS1 ribosomal DNA marker

“…The morphology of F. hepatica and F. gigantica can generally be used for distinguishing between them, but for detecting and distinguishing intermediate forms between Fasciola species, one need to use molecular methods, and genetic markers (20). Aryaeipour et al (31) concluded that the PCR-RFLP method was a more reliable method than morphology for the differentiation of Fasciola species and the morphological methods were inefficient for determining genetic diversity. The first and the second internal transcribed spacers (ITS1 and ITS2) of the nuclear ribosomal DNA (rDNA) were used as genetic markers for genetic characterization and differential between the species of Fasciola that have been confirmed by several previous studies (32)(33)(34).…”

“…These markers are non-coding regions between the 18S, 5.8S, and 28S coding regions and have been used for diagnostic purposes at the species level (35)(36)(37)(38). In the Middle East, many studies relied on the ITS1 marker to identify Fasciola species (39,40). In this research, Fasciola spp.…”

Assessment of genetic markers for multilocus sequence typing (MLST) of Fasciola isolates from Iran

“…Several markers have been developed and used to assess the population structure of Fasciola species such as the ribosomal internal transcribed spacer (ITS), phosphoenolpyruvate carboxykinase (Pepck), DNA polymerase delta (Pold), cytochrome c oxidase I (COI) and the NADH dehydrogenase (ND) genes (Bozorgomid et al, 2020;Heydarian et al, 2022;Kasahara et al, 2021;Shafiei et al, 2014). It has been reported that the genetic diversity of this parasite may have a role in drug resistance, virulence, pathogenicity, clinical characteristics of the disease and certain epidemiological features (Lalor et al, 2021).…”

“…F. hepatica and F. gigantica can generally be distinguished on the basis of morphological criteria but the existence of parthenogenic (aspermic) Fasciola flukes with intermediate morphological characteristics between the two species can cause confusion (Aryaeipour et al., 2017 ). Several markers have been developed and used to assess the population structure of Fasciola species such as the ribosomal internal transcribed spacer (ITS), phosphoenolpyruvate carboxykinase ( Pepck ), DNA polymerase delta ( Pold ), cytochrome c oxidase I ( COI ) and the NADH dehydrogenase ( ND ) genes (Bozorgomid et al., 2020 ; Heydarian et al., 2022 ; Kasahara et al., 2021 ; Shafiei et al., 2014 ). It has been reported that the genetic diversity of this parasite may have a role in drug resistance, virulence, pathogenicity, clinical characteristics of the disease and certain epidemiological features (Lalor et al., 2021 ).…”