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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

Location of the sites where Fasciola isolates were collected in West Azerbaijan Province, Iran
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Figure 1: Location of the sites where Fasciola isolates were collected in West Azerbaijan Province, Iran

Mentions: Adult trematodes (n=580) were collected from the 90 liver of slathered cattles (n=335) and sheep (n=245) in Salmas (n=65 cattle, 45 sheep), Makou (n=50 cattle, 50 sheep), Urmia (n=90 cattle, 70 sheep), Mehabad (n=80 cattle, 50 sheep), and Bukan (n=50 cattle, 30 sheep) districts as well as West Azerbaijan Province, Iran (Fig. 1). All samples were washed in normal saline, fixed in 70% ethanol, and then kept at room temperature until DNA extraction. All morphological measurements of adults were made according to methods described for Fasciola (9, 21, 31). The morphometric values such as, body length (BL), body width (BW), cephalic cone length (CL) and length of area behind the testes, were obtained using a microscope and calibrated ocular micrometer.


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)

Location of the sites where Fasciola isolates were collected in West Azerbaijan Province, Iran
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4835470&req=5

Figure 1: Location of the sites where Fasciola isolates were collected in West Azerbaijan Province, Iran
Mentions: Adult trematodes (n=580) were collected from the 90 liver of slathered cattles (n=335) and sheep (n=245) in Salmas (n=65 cattle, 45 sheep), Makou (n=50 cattle, 50 sheep), Urmia (n=90 cattle, 70 sheep), Mehabad (n=80 cattle, 50 sheep), and Bukan (n=50 cattle, 30 sheep) districts as well as West Azerbaijan Province, Iran (Fig. 1). All samples were washed in normal saline, fixed in 70% ethanol, and then kept at room temperature until DNA extraction. All morphological measurements of adults were made according to methods described for Fasciola (9, 21, 31). The morphometric values such as, body length (BL), body width (BW), cephalic cone length (CL) and length of area behind the testes, were obtained using a microscope and calibrated ocular micrometer.

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