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Genetic Characterization of Fasciola Isolates from West Azerbaijan Province Iran Based on ITS1 and ITS2 Sequence of Ribosomal DNA.

Galavani H, Gholizadeh S, Hazrati Tappeh K - Iran J Parasitol (2016 Jan-Mar)

Bottom Line: PCR amplification of 1081 bp fragment and sequencing result showed 100% similarity with F. hepatica in ITS1 (428 bp), 5.8s (158 bp), and ITS2 (366 bp) regions.However, 5 and 6 bp variation in ITS1 and ITS2 regions, respectively, is not enough to separate of Fasciola spp.Therefore, more studies are essential for designing new molecular markers to correct species identification.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology and Mycology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.

ABSTRACT

Background: Fascioliasis, caused by Fasciola hepatica and F. gigantica, has medical and economic importance in the world. Molecular approaches comparing traditional methods using for identification and characterization of Fasciola spp. are precise and reliable. The aims of current study were molecular characterization of Fasciola spp. in West Azerbaijan Province, Iran and then comparative analysis of them using GenBank sequences.

Methods: A total number of 580 isolates were collected from different hosts in five cities of West Azerbaijan Province, in 2014 from 90 slaughtered cattle (n=50) and sheep (n=40). After morphological identification and DNA extraction, designing specific primer were used to amplification of ITS1, 5.8s and ITS2 regions, 50 samples were conducted to sequence, randomly.

Result: Using morphometric characters 99.14% and 0.86% of isolates identified as F. hepatica and F. gigantica, respectively. PCR amplification of 1081 bp fragment and sequencing result showed 100% similarity with F. hepatica in ITS1 (428 bp), 5.8s (158 bp), and ITS2 (366 bp) regions. Sequence comparison among current study sequences and GenBank data showed 98% identity with 11 nucleotide mismatches. However, in phylogenetic tree F. hepatica sequences of West Azerbaijan Province, Iran, were in a close relationship with Iranian, Asian, and African isolates.

Conclusions: Only F. hepatica species is distributed among sheep and cattle in West Azerbaijan Province Iran. However, 5 and 6 bp variation in ITS1 and ITS2 regions, respectively, is not enough to separate of Fasciola spp. Therefore, more studies are essential for designing new molecular markers to correct species identification.

No MeSH data available.


Related in: MedlinePlus

Multiple sequence alignments of the rDNA ITS1, 5.8s, and ITS2 regions of F. hepatica (JF708027) and F. gigantica (JF432073) as well as representative F. hepatica sequences of the current study [KF531639 (ITS1), KF531689 (5.8s), KF531739 (ITS2)]. Bold sequences belong to 5.8s region flanking to ITS1 and ITS2
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Figure 2: Multiple sequence alignments of the rDNA ITS1, 5.8s, and ITS2 regions of F. hepatica (JF708027) and F. gigantica (JF432073) as well as representative F. hepatica sequences of the current study [KF531639 (ITS1), KF531689 (5.8s), KF531739 (ITS2)]. Bold sequences belong to 5.8s region flanking to ITS1 and ITS2

Mentions: The sequence analysis showed that ITS1, 5.8s, and ITS2 had a length of 428, 158, and 366 bp, i.e. 952 bp in total (Fig. 2). The BLAST analysis of rDNA ITS1, 5.8s, and ITS2 sequences showed 100% similarity with F. hepatica and 98% with F. gigantic (Fig. 2). There were 11 mismatches in positions 24, 114, 208, 286, 306, 821, 860, 866, 918, 924, and 938, including seven transitions, two transversions, one insertion, and one deletion (Fig. 2). The sequence compositions of ITS1, 5.8s, and ITS2 regions were 51.87% (GC) and 48.13% (AT), 53.17% (GC) and 46.83% (AT) as well as 48.63% (GC) and 51.37% (AT), respectively. Based on morphological characteristics, 0.86% (5 samples) was identified as F. gigantica. In addition, ITS1, 5.8s, and ITS2 sequences of these samples showed 100% similarity with F. hepatica (Fig. 2).


Genetic Characterization of Fasciola Isolates from West Azerbaijan Province Iran Based on ITS1 and ITS2 Sequence of Ribosomal DNA.

Galavani H, Gholizadeh S, Hazrati Tappeh K - Iran J Parasitol (2016 Jan-Mar)

Multiple sequence alignments of the rDNA ITS1, 5.8s, and ITS2 regions of F. hepatica (JF708027) and F. gigantica (JF432073) as well as representative F. hepatica sequences of the current study [KF531639 (ITS1), KF531689 (5.8s), KF531739 (ITS2)]. Bold sequences belong to 5.8s region flanking to ITS1 and ITS2
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4835470&req=5

Figure 2: Multiple sequence alignments of the rDNA ITS1, 5.8s, and ITS2 regions of F. hepatica (JF708027) and F. gigantica (JF432073) as well as representative F. hepatica sequences of the current study [KF531639 (ITS1), KF531689 (5.8s), KF531739 (ITS2)]. Bold sequences belong to 5.8s region flanking to ITS1 and ITS2
Mentions: The sequence analysis showed that ITS1, 5.8s, and ITS2 had a length of 428, 158, and 366 bp, i.e. 952 bp in total (Fig. 2). The BLAST analysis of rDNA ITS1, 5.8s, and ITS2 sequences showed 100% similarity with F. hepatica and 98% with F. gigantic (Fig. 2). There were 11 mismatches in positions 24, 114, 208, 286, 306, 821, 860, 866, 918, 924, and 938, including seven transitions, two transversions, one insertion, and one deletion (Fig. 2). The sequence compositions of ITS1, 5.8s, and ITS2 regions were 51.87% (GC) and 48.13% (AT), 53.17% (GC) and 46.83% (AT) as well as 48.63% (GC) and 51.37% (AT), respectively. Based on morphological characteristics, 0.86% (5 samples) was identified as F. gigantica. In addition, ITS1, 5.8s, and ITS2 sequences of these samples showed 100% similarity with F. hepatica (Fig. 2).

Bottom Line: PCR amplification of 1081 bp fragment and sequencing result showed 100% similarity with F. hepatica in ITS1 (428 bp), 5.8s (158 bp), and ITS2 (366 bp) regions.However, 5 and 6 bp variation in ITS1 and ITS2 regions, respectively, is not enough to separate of Fasciola spp.Therefore, more studies are essential for designing new molecular markers to correct species identification.

View Article: PubMed Central - PubMed

Affiliation: Department of Parasitology and Mycology, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran.

ABSTRACT

Background: Fascioliasis, caused by Fasciola hepatica and F. gigantica, has medical and economic importance in the world. Molecular approaches comparing traditional methods using for identification and characterization of Fasciola spp. are precise and reliable. The aims of current study were molecular characterization of Fasciola spp. in West Azerbaijan Province, Iran and then comparative analysis of them using GenBank sequences.

Methods: A total number of 580 isolates were collected from different hosts in five cities of West Azerbaijan Province, in 2014 from 90 slaughtered cattle (n=50) and sheep (n=40). After morphological identification and DNA extraction, designing specific primer were used to amplification of ITS1, 5.8s and ITS2 regions, 50 samples were conducted to sequence, randomly.

Result: Using morphometric characters 99.14% and 0.86% of isolates identified as F. hepatica and F. gigantica, respectively. PCR amplification of 1081 bp fragment and sequencing result showed 100% similarity with F. hepatica in ITS1 (428 bp), 5.8s (158 bp), and ITS2 (366 bp) regions. Sequence comparison among current study sequences and GenBank data showed 98% identity with 11 nucleotide mismatches. However, in phylogenetic tree F. hepatica sequences of West Azerbaijan Province, Iran, were in a close relationship with Iranian, Asian, and African isolates.

Conclusions: Only F. hepatica species is distributed among sheep and cattle in West Azerbaijan Province Iran. However, 5 and 6 bp variation in ITS1 and ITS2 regions, respectively, is not enough to separate of Fasciola spp. Therefore, more studies are essential for designing new molecular markers to correct species identification.

No MeSH data available.


Related in: MedlinePlus