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. 2014:2014:405740.
doi: 10.1155/2014/405740. Epub 2014 Jun 11.

Molecular and Morphological Characterization of Fasciola spp. Isolated from Different Host Species in a Newly Emerging Focus of Human Fascioliasis in Iran

Reza Shafiei  1 , Bahador Sarkari  1 , Seyed Mahmuod Sadjjadi  1 , Gholam Reza Mowlavi  2 , Abdolali Moshfe  3
Affiliations

Molecular and Morphological Characterization of Fasciola spp. Isolated from Different Host Species in a Newly Emerging Focus of Human Fascioliasis in Iran

Reza Shafiei et al. Vet Med Int. 2014.

Abstract

The current study aimed to find out the morphometric and genotypic divergences of the flukes isolated from different hosts in a newly emerging focus of human fascioliasis in Iran. Adult Fasciola spp. were collected from 34 cattle, 13 sheep, and 11 goats from Kohgiluyeh and Boyer-Ahmad province, southwest of Iran. Genomic DNA was extracted from the flukes and PCR-RFLP was used to characterize the isolates. The ITS1, ITS2, and mitochondrial genes (mtDNA) of NDI and COI from individual liver flukes were amplified and the amplicons were sequenced. Genetic variation within and between the species was evaluated by comparing the sequences. Moreover, morphometric characteristics of flukes were measured through a computer image analysis system. Based on RFLP profile, from the total of 58 isolates, 41 isolates (from cattle, sheep, and goat) were identified as Fasciola hepatica, while 17 isolates from cattle were identified as Fasciola gigantica. Comparison of the ITS1 and ITS2 sequences showed six and seven single-base substitutions, resulting in segregation of the specimens into two different genotypes. The sequences of COI markers showed seven DNA polymorphic sites for F. hepatica and 35 DNA polymorphic sites for F. gigantica. Morphological diversity of the two species was observed in linear, ratios, and areas measurements. The findings have implications for studying the population genetics, epidemiology, and control of the disease.

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Figures

Figure 1
Figure 1
Map of Iran (a) and the Kohgiluyeh and Boyer-Ahmad province (b).
Figure 2
Figure 2
Fasciola hepatica (a) isolated from sheep and Fasciola gigantica (b) isolated from cattle.
Figure 3
Figure 3
Alignment of ITS1 sequence of F. hepatica (AB207139) and F. gigantica (AB207142), with F. gigantica and F. hepatica of Kohgiluyeh and Boyer-Ahmad province, Iran.
Figure 4
Figure 4
Alignment of ITS2 sequence of F. gigantica (AB010977) and F. hepatica (AB207148), with F. gigantica and F. hepatica of Kohgiluyeh and Boyer-Ahmad province, Iran.
Figure 5
Figure 5
Phylogenetic relationship of ITS1 sequences of isolates of Fasciola hepatica and Fasciola gigantica from Iran using Maximum Likelihood method. Fascioloides magna (AN: EF051080) was used as the out group.
Figure 6
Figure 6
Phylogenetic relationship of ITS2 sequences of isolates of Fasciola hepatica and Fasciola gigantica from Iran using Maximum Likelihood method. Fascioloides magna (AN: EF534995) was used as the out group.
Figure 7
Figure 7
Phylogenetic relationship of COI sequences of isolates of Fasciola hepatica and Fasciola gigantica from Iran using Maximum Likelihood method. Paragonimus westermani (AN: AF219379) was used as the out group.
Figure 8
Figure 8
Phylogenetic relationship of NDI sequences of isolates of Fasciola hepatica and Fasciola gigantica from Iran using Maximum Likelihood method. Paragonimus westermani (AN: AF219379) was used as the out group.
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