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Morphological and molecular discrimination of fasciola species isolated from domestic ruminants of urmia city, iran.

Yakhchali M, Malekzadeh-Viayeh R, Imani-Baran A, Mardani K - Iran J Parasitol (2015 Jan-Mar)

Bottom Line: While the phylogenetic reconstruction justified, to some extent, the morphological diagnosis, it failed to segregate F. hepatica from F. gigantica identified in this and the previous studies.To resolve fully the problem of taxonomy and evolution in Fasciola species, employing a broad range of molecular and morphological approaches is necessary.This is crucial for epidemiological surveys and successful clinical management of their infection.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.

ABSTRACT

Background: The trematodes of the genus Fasciola (the liver flukes) are among the well-known instances of food-borne parasites worldwide. Differentiation of Fasciola species is important because of their different transmission and epidemiological characteristics. The current study was undertaken to discriminate Fasciola species in the domestic ruminants of Urmia city, Iran.

Methods: Adult flukes were isolated from the naturally infected livers of the slaughtered water buffaloes and sheep. The flukes were initially identified based on morphological and morphometric parameters. A 618-bp-long fragment of the 28SrRNA gene of Fasciola was amplified by polymerase chain reaction (PCR). The amplified fragment was digested by DraII or AvaII enzymes for a restriction fragment length polymorphism (RFLP) analysis and sequenced for the phylogenetic tree construction.

Results: Based on the morphometric examination, the flukes belonged to F. hepatica, F. gigantica and an intermediate Fasciola form. The PCR-RFLP analysis was able to differentiate F. hepatica from F. gigantica. While the phylogenetic reconstruction justified, to some extent, the morphological diagnosis, it failed to segregate F. hepatica from F. gigantica identified in this and the previous studies.

Conclusion: To resolve fully the problem of taxonomy and evolution in Fasciola species, employing a broad range of molecular and morphological approaches is necessary. This is crucial for epidemiological surveys and successful clinical management of their infection.

No MeSH data available.


Related in: MedlinePlus

Restriction fragment length polymorphism (RFLP) pattern of the PCR products of the liver flukes after digestion with DraII restriction enzyme. Lanes 1–7, PCR products of F. hepatica; Lanes 9–16, PCR products of F. gigantica. Lane P: 618-bp-long PCR product of F. gigantica; Lane M, 250bp DNA size marker
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Figure 3: Restriction fragment length polymorphism (RFLP) pattern of the PCR products of the liver flukes after digestion with DraII restriction enzyme. Lanes 1–7, PCR products of F. hepatica; Lanes 9–16, PCR products of F. gigantica. Lane P: 618-bp-long PCR product of F. gigantica; Lane M, 250bp DNA size marker

Mentions: Based on the RFLP analysis, restriction of the PCR products by the enzyme AvaII produced two fragments of 529 and 62bp for F. hepatica and 322 and 269bp for F. gigantica isolates (Fig. 2). DraII enzyme digested the PCR product of F. hepatica in one position generating a fragment of 529bp in length, while the enzyme was unable to digest the PCR product of F. gigantica (Fig. 3).


Morphological and molecular discrimination of fasciola species isolated from domestic ruminants of urmia city, iran.

Yakhchali M, Malekzadeh-Viayeh R, Imani-Baran A, Mardani K - Iran J Parasitol (2015 Jan-Mar)

Restriction fragment length polymorphism (RFLP) pattern of the PCR products of the liver flukes after digestion with DraII restriction enzyme. Lanes 1–7, PCR products of F. hepatica; Lanes 9–16, PCR products of F. gigantica. Lane P: 618-bp-long PCR product of F. gigantica; Lane M, 250bp DNA size marker
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4403539&req=5

Figure 3: Restriction fragment length polymorphism (RFLP) pattern of the PCR products of the liver flukes after digestion with DraII restriction enzyme. Lanes 1–7, PCR products of F. hepatica; Lanes 9–16, PCR products of F. gigantica. Lane P: 618-bp-long PCR product of F. gigantica; Lane M, 250bp DNA size marker
Mentions: Based on the RFLP analysis, restriction of the PCR products by the enzyme AvaII produced two fragments of 529 and 62bp for F. hepatica and 322 and 269bp for F. gigantica isolates (Fig. 2). DraII enzyme digested the PCR product of F. hepatica in one position generating a fragment of 529bp in length, while the enzyme was unable to digest the PCR product of F. gigantica (Fig. 3).

Bottom Line: While the phylogenetic reconstruction justified, to some extent, the morphological diagnosis, it failed to segregate F. hepatica from F. gigantica identified in this and the previous studies.To resolve fully the problem of taxonomy and evolution in Fasciola species, employing a broad range of molecular and morphological approaches is necessary.This is crucial for epidemiological surveys and successful clinical management of their infection.

View Article: PubMed Central - PubMed

Affiliation: Dept. of Pathobiology, Faculty of Veterinary Medicine, Urmia University, Urmia, Iran.

ABSTRACT

Background: The trematodes of the genus Fasciola (the liver flukes) are among the well-known instances of food-borne parasites worldwide. Differentiation of Fasciola species is important because of their different transmission and epidemiological characteristics. The current study was undertaken to discriminate Fasciola species in the domestic ruminants of Urmia city, Iran.

Methods: Adult flukes were isolated from the naturally infected livers of the slaughtered water buffaloes and sheep. The flukes were initially identified based on morphological and morphometric parameters. A 618-bp-long fragment of the 28SrRNA gene of Fasciola was amplified by polymerase chain reaction (PCR). The amplified fragment was digested by DraII or AvaII enzymes for a restriction fragment length polymorphism (RFLP) analysis and sequenced for the phylogenetic tree construction.

Results: Based on the morphometric examination, the flukes belonged to F. hepatica, F. gigantica and an intermediate Fasciola form. The PCR-RFLP analysis was able to differentiate F. hepatica from F. gigantica. While the phylogenetic reconstruction justified, to some extent, the morphological diagnosis, it failed to segregate F. hepatica from F. gigantica identified in this and the previous studies.

Conclusion: To resolve fully the problem of taxonomy and evolution in Fasciola species, employing a broad range of molecular and morphological approaches is necessary. This is crucial for epidemiological surveys and successful clinical management of their infection.

No MeSH data available.


Related in: MedlinePlus