Identification and Phylogenetic Classification of Fasciola species Isolated from Sheep and Cattle by PCR-RFLP in Zabol, in Sis-tan and Baluchistan Province, Southeast Iran

Mir, Sedighe; Dabirzadeh, Mansour; Rokni, Mohammad Bagher; Aryaeipour, Mojgan; Shahraki, Mahdi Khoshsima; Azizi, Hakim

doi:10.18502/ijph.v48i5.1811

Cited by 8 publications

(8 citation statements)

References 19 publications

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“…The phylogenetic tree showed that the Fasciola flukes of this study were scattered as pure F. hepatica and F. gigantica clades, proposing that two genotypes of Fasciola can infect ruminants in Duhok province, these results agree with those of Shafiei et al (2014) and Mir et al (2019) in Iran. Furthermore, the present results showed the presence of a higher genetic diversity of the nuclear ITS2 sequence between F. hepatica and F. gigantica than that between them and F. nyanzea (Dinnik and Dinnik 2009) and both Fasciola species (F. hepatica and F. gigantica).…”

Section: Resultssupporting

confidence: 92%

“…However, the results of the studies conducted by Peng et al (2009), Nguyen et al (2018), andHaridwal et al (2021) failed to distinguish between the two species of Fasciola (F. hepatica and F. gigantica) using the morphometric characters, due to the presence of intermediate forms. Furthermore, in areas where the possibility of the existence of intermediate and hybrid forms exists, it is necessary to use molecular characterization and evaluation to distinguish between species (Mir et al, 2019, Karimi, 2008, Saki and Yousefi, 2011. In this study, the adult flukes that look like hybrid forms are hardly distinguishable by morphological methods; therefore, they must be distinguished by molecular approaches as described by Ai et al (2011).…”

Section: Resultsmentioning

confidence: 99%

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Molecular Identification And Phylogenetic Analysis of Fasciola Species Isolated from Sheep And Cattle Using The Its2 Rdna Marker in Duhok Province, Kurdistan Region, Iraq

Rekani

Mero²

2022

UOD

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The aims of the current study were molecular identification and phylogenetic analysis of Fasciola spp. isolated from infected cattle and sheep using the ITS2 region of rDNA as a molecular marker in Duhok province, Kurdistan region, Iraq. DNA was extracted from 40 adult flukes isolated from the livers of infected sheep (23) and cattle (17) slaughtered at the abattoirs of Duhok Province. The amplification of the rDNA ITS2 region of these 40 specimens was done by PCR using primers (FITS2-F and FITS2-R) designed for this study by the authors. ITS2 fragments (approximately 314 bp) of the rDNA were successfully amplified, and produced monoclear bands, that were confirmed by gel electrophoresis. From these isolates, 5 amplicons of ITS2 (2 sheep and 3 cattle) were selected for sequencing, and then the obtained sequences were compared to those in NCBI-GenBank for genotyping and phylogenetic analysis. The blast alignment of NCBI-GenBank revealed four sequences belonging to Fasciola hepatica and one belonging to Fasciola gigantica, both with 100% similarity. The genetic distance between F. hepatica and F. gigantica was 0.0127. Phylogenetic analysis revealed that the sequences in this study were clustered into two main clades (F. hepatica and F. gigantica), with other sequences available in GenBank. The study concluded that ITS2 rDNA sequences were appropriate to identify both Fasciola species isolated from the infected sheep and cattle in Duhok province, which were identical to those of neighboring countries (Iran and Turkey). The primers of ITS2 rDNA region that designed for the current study can be useful for the diagnosis of animals and humans fasciolosis.

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Molecular Identification And Phylogenetic Analysis of Fasciola Species Isolated from Sheep And Cattle Using The Its2 Rdna Marker in Duhok Province, Kurdistan Region, Iraq

Rekani

Mero²

2022

UOD

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“…Alignment analysis revealed very close similarity among the nucleotide sequences, and genetic variation was not observed within the same species (no intraspecific variation within species) of the current study and BLAST hits in GenBank. On the other hand, significant genetic variability was detected between F. hepatica and F. gigantica, and these results agree with those of Shaldoum et al (51) and Mir et al (2), which used the same marker (ITS1) to distinguish between the two Fasciola species.…”

Section: Discussionsupporting

confidence: 91%

“…Results were also identical to those from Iran, Pakistan, India, China, Nigeria, and Kenya for F. gigantica as reported by Hamoo et al (53) in Duhok province (in Aqrah city), and also this close relationship between the same species of Fasciola has been shown by Nguyen et al (54). The analysis of the rDNA ITS1 sequence of Fasciola species was commonly reported in Asia, Europe, and African countries (2,18,49). The present study results and those of other studies demonstrate the diagnostic power of the ITS1 region in differentiating the Fasciola species (39,55).…”

Section: Discussionsupporting

confidence: 87%

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

Rekani¹,

Mero²

2023

IJVS

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This study aimed to characterize and identify the genotypes of Fasciola spp. isolated from sheep, goats, and cattle in Duhok province based on the ITS1 region of rDNA. About 54 adult Fasciola flukes were individually isolated from the livers of naturally infected ruminants. After morphological identification, the genomic DNA of 54 isolated Fasciola spp. was successfully extracted, and the ITS1segment (518 bp) of rDNA was amplified. The amplicons were confirmed by gel electrophoresis and yielded mono cleared bands. Five amplicons from these samples (2 sheep, 2 cattle, and 1 goat) were selected for sequencing and then compared with NCBI-GenBank sequences for genotyping and phylogenetic analysis. Sequencing analysis and the BLAST results revealed that 3/5 of the resultant sequences were F. hepatica and 2/5 were F. gigantica. The ITS1 sequences were submitted to NCBI-GenBank with accession numbers: OM920533, OM920534, OM948733, OM948683, and OM918714. Alignment analysis of the current study and GenBank ITS1 sequences showed the presence of nucleotide variations between F. hepatica and F. gigantica species (interspecific), which were enough to separate them. At the same time, they were not observed within the same species of Fasciola (intraspecific). The pairwise identity percentage of intraspecific and interspecific Fasciola isolates was 100% and 99.2-99.6%, respectively. Phylogenetic analysis of the ITS1 sequences demonstrated that the Fasciola isolates of this study were clustered into two clades (hepatica and gigantica clades). The present study concluded that both Fasciola spp. (F. hepatica and F. gigantica) existed among the infected ruminants in Duhok province, and are closely related to intraspecific Fasciola isolates from different countries in the Middle East, Asia, Europe, and Africa.

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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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Identification and Phylogenetic Classification of Fasciola species Isolated from Sheep and Cattle by PCR-RFLP in Zabol, in Sis-tan and Baluchistan Province, Southeast Iran

Cited by 8 publications

References 19 publications

Molecular Identification And Phylogenetic Analysis of Fasciola Species Isolated from Sheep And Cattle Using The Its2 Rdna Marker in Duhok Province, Kurdistan Region, Iraq

Molecular Identification And Phylogenetic Analysis of Fasciola Species Isolated from Sheep And Cattle Using The Its2 Rdna Marker in Duhok Province, Kurdistan Region, Iraq

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

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

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