Novel methods for the molecular discrimination of Fasciola spp. on the basis of nuclear protein-coding genes

Shoriki, Takuya; Ichikawa‐Seki, Madoka; Suganuma, Keisuke; Naito, Ikunori; Hayashi, Kei; Nakao, Minoru; Aita, Junya; Mohanta, Uday Kumar; Inoue, Noboru; Murakami, Kenji; Itagaki, Tadashi

doi:10.1016/j.parint.2015.12.002

Cited by 48 publications

(40 citation statements)

References 20 publications

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“…All 60 Fasciola flukes collected from western Java in Indonesia were identified as F. gigantica in an analysis of the nuclear pepck and pold genes (Table 1) [5]. These results are consistent with the nad1 haplotypes insofar as all the flukes had haplotypes belonging to the F. gigantica haplogroup based on the nad1 gene ( Fig.…”

Section: Discussionsupporting

confidence: 71%

“…Therefore, all the Fasciola flukes examined in this study were identified as F. gigantica. None of the western Java samples showed the mixed fragment pattern of hybrid Fasciola flukes [5].…”

Section: Species Identificationmentioning

confidence: 96%

“…These flukes displayed the F. gigantica fragment patterns for both the pepck and pold genes [5]. The remaining 16 flukes contained no sperm in their seminal vesicles, but they also displayed the F. gigantica fragment patterns for the pepck and pold genes.…”

Section: Species Identificationmentioning

confidence: 99%

“…The pepck region was amplified with multiplex PCR using primers Fhpepck-F, Fg-pepck-F, and Fcmn-pepck-R [5], and the fragment patterns were compared on 1.0% agarose gels. The pold gene was analyzed with PCR-restriction fragment length polymorphism [5]. Briefly, the pold gene was amplified with primers Fasciola-pold-F1 and Fasciola-pold-R1.…”

Section: Dna Extraction and Molecular Species Identificationmentioning

confidence: 99%

“…In addition to the two species, aspermic Fasciola flukes, which have few or no spermatozoa in their seminal vesicles, have been reported in Asian countries [4,7]. Recently, novel nuclear markers, the phosphoenolpyruvate carboxykinase (pepck) and DNA polymerase delta (pold) genes, have been used for the precise discrimination of F. hepatica, F. gigantica, and aspermic Fasciola flukes [5], which can be clearly distinguished with a multiplex polymerase chain reaction (PCR) and/or a PCR-restriction fragment length polymorphism method. By using these methods, aspermic Fasciola flukes display a mixed fragment pattern combining those of the two species, and are therefore thought to be their descendants, generated by hybridization between F. hepatica and F. gigantica [5].…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia

Ichikawa‐Seki

Allamanda²,

Wibowo³

et al. 2016

Parasitology International

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Section: Discussionsupporting

confidence: 71%

Section: Species Identificationmentioning

confidence: 96%

Section: Species Identificationmentioning

confidence: 99%

Section: Dna Extraction and Molecular Species Identificationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia

Ichikawa‐Seki

Allamanda²,

Wibowo³

et al. 2016

Parasitology International

Self Cite

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Morphological and molecular characterization of Fasciola isolates from livestock in Golestan province, northern Iran

Sharbatkhori

Nasibi

Mohammadi

et al. 2023

Veterinary Medicine & Sci

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Background: Fascioliasis, caused by the liver flukes Fasciola hepatica and Fasciola gigantica, is a global zoonotic helminthic disease. The livestock and human are the final hosts of the parasites. Northern Iran is an important endemic region for fascioliasis. Few studies have been conducted on the characterization of Fasciola isolates from eastern regions of the Caspian littoral of the country. Objective:The aim of the present study was to identify F. hepatica, F. gigantica and intermediate/hybrid forms of Fasciola isolates from livestock in Golestan province, northern Iran, using morphometric and molecular tools.Methods: Livestock livers naturally infected with Fasciola spp. were collected from Golestan slaughterhouse during 2019-2020. The worms were morphometrically studied using a calibrated stereomicroscope. Genomic DNA was extracted from all samples, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed on internal transcribed spacer (ITS1) region using Rsa1 restriction enzyme. All the isolates were then analysed by multiplex PCR on Pepck region.Results: A total of 110 Fasciola isolates were collected from the infected livers, including 94 sheep, 12 cattle and 4 goats. Morphometric analysis of 61 adult Fasciola isolates indicated that, 44 and 17 isolates belonged to F. hepatica and F. gigantica, respectively.Eighty-one and 29 isolates belonged to F. hepatica and F. gigantica using ITS1-RFLP, respectively. However, Pepck Multiplex PCR indicated 72 F. hepatica, 26 F. gigantica and 12 intermediate/hybrid forms. All 12 hybrid isolates were found in sheep host. Two isolates were identified as F. gigantica using morphometry and F. hepatica using both molecular methods. Conclusion:The present study confirmed the existence of both F. hepatica and F. gigantica species and reported the first molecular evidence of hybrid Fasciola isolates in ruminants of Golestan province.

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Morphometric and Molecular Characterization of Fasciola spp. in Livestock From Northwestern Provinces of Iran

Galavani,

Raeghi,

Karamian

et al. 2024

Veterinary Medicine & Sci

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BackgroundLiver flukes from the genus Fasciola are the causative agents for human and livestock fascioliasis. Accurate identification of Fasciola spp. is essential to understanding the epidemiology of fascioliasis. This study aimed to determine the morphometric and molecular characterization of Fasciola spp. in livestock from Northwestern provinces of Iran.MethodsFive hundred adult Fasciola flukes were obtained from different definitive hosts (cattle, sheep, goats, and buffaloes) in four local abattoirs in the northwestern provinces of Iran (West‐Azerbaijan, East‐Azerbaijan, Ardabil, and Zanjan) from September 2021 to August 2022. All samples were examined by morphometric criteria; then, 49 samples were identified using PCR‐RFLP based on ITS1 region and 23 sequence of isolates analyzed by cox1 marker. PCR‐RFLP methods compared morphometric results, and cox1 gene sequences were used to confirm PCR‐RFLP results and phylogenetic analysis.ResultsThe differences between the body length, body width, cephalic cone length, cephalic cone width, body area, and distance between the ventral sucker and posterior end of the body in two species were significant (p < 0.05). Based on the morphometric criteria, 139 samples (27.8%) were identified as Fasciola gigantica and 361 (72.2%) as Fasciola hepatica. Similarly, PCR‐RFLP analysis of ITS1 region confirmed morphometric results. No hybrid forms of Fasciola were detected. Partial sequences of cox1 showed 13 variable sites with eight haplotypes in F. hepatica and 12 variable sites with five haplotypes in F. gigantica.ConclusionBased on the results of this study, the PCR‐RFLP method can be used to confirm the morphological method of Fasciola species, but it is insufficient to study their genetic diversity. Also, sequences of cox1 results of the present study showed that F. hepatica and F. gigantica species in the Northern provinces of Iran have different genetic structures and haplotypes.

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Novel methods for the molecular discrimination of Fasciola spp. on the basis of nuclear protein-coding genes

Cited by 48 publications

References 20 publications

Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia

Molecular characterization and phylogenetic analysis of Fasciola gigantica from western Java, Indonesia

Morphological and molecular characterization of Fasciola isolates from livestock in Golestan province, northern Iran

Morphometric and Molecular Characterization of Fasciola spp. in Livestock From Northwestern Provinces of Iran

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