Iranian Journal of Parasitology 2015. 10(1):46-55.

Morphological and molecular discrimination of fasciola species isolated from domestic ruminants of urmia city, iran.
Mohammad Yakhchali, Reza Malekzadeh-Viayeh, Abbas Imani-Baran, Karim Mardani

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 epidemio-logical 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 frag-ment length polymorphism (RFLP) analysis and sequenced for the phylogenetic tree construction.

Results:Based on the morphometric examination, the flukes belonged to F. he-patica, F. gigantica and an intermediate Fasciola form. The PCR-RFLP analysis was able to differentiate F. hepatica from F. gigantica. While the phylogenetic reconstruc-tion 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 man-agement of their infection.


Keywords


28SrRNA gene; Liver flukes; Morphology; PCR-sequencing; RFLP

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References


Mas-Coma S, Bargues MD, Valero MA. Fascio-liasis and other plant-borne trematode zoonoses. Int J Parasitol. 2005; 35:1255-78. 2. Lin RQ, Dong SJ, Nie K, Wang CR, Song HQ, Li AX, Huang WY, Zhu XQ. Sequence analysis of the first internal transcribed spacer of rDNA supports the existence of the intermediate Fasciola between F. hepatica and F. gigantica in mainland China. Parasitol Res. 2007; 101(3): 813-7.

Mas-Coma S. Human fascioliasis. In: Waterborne Zoonoses: Identification, Causes and Control. Cotruvo JA, Dufour A, Rees G, Bartram J, Carr R, Cliver DO, Craun GF, Fayer R, Gannon VPJ, editors. World Health Organization (WHO), London, UK: IWA Publishing; 2004.

Raina OK, Jacob SS, Sankar M, Bhattacharya D, Bandyopadyay S, Varghese A, Chamuah JK, Lal-rinkima H. Genetic characterization of Fasciola gi-gantica from different geographical regions of In-dia by ribosomal DNA markers. J Parasit Dis. 2013; doi: 10.1007/s12639.013.0276.7.

Massoud J. Fascioliasis outbreak of man and drug test (Triclabendazol) in Caspian littoral, northern part of Iran, 1989. Bull Soc Fr Parasitol. 1990; 8 (Suppl 1): 438.

Pourtaghva M, Shafi A, Saberi A, Bahar K, So-laymanlou F. Fasciolase en Iran. Bull Soc Fr Para-sitol. 1990; 8(Suppl. 1): 404.

Yadegari D, Forghanparast K, Assmar M. Inves-tigation of an epidemic of fascioliasis in north Iran. Bull Soc Fr Parasitol. 1990; 8 (Suppl2): 868.

Assmar M, Milaninia A, Amir-Khani A, Yadegari AD, Forghanparast K, Nahravanian H, Piazak N, Esmayli A, Hovanesian A, Valadkhani Z. Seroep-idemiological investigation of fascioliasis in north-ern Iran. Medical Journal of the Islamic Republic of Iran 1991; 5, 23-7.

Forghanparast K. Clinicoepidemiological study of human fascioliasis in Guilan. Journal of the Gui-lan University of Medical Sciences. 1993; 2: 63-4.

Yakhchali M, Ghobadi K. A survey on hel-minthes infection of liver and economic loss in slaughtered sheep of Urmia slaughterhouse, Iran. Iranian Veterinary Journal. 2005; 11: 60-5. (in Per-sian with English abstract)

Mas-Coma S, Funatsu IR, Bargues MD. Fasciola hepatica and Lymnaeid snails occurring at very high altitude in South America. Parasitol. 2001; 123: 115-27.

Periago MV, Valero MA, El Sayed M, Ashrafi K, El Wakeel A, Mohamed MY, Desquesnes M, Curtale F, Mas-Coma S. First phenotypic descrip-tion of Fasciola hepatica/Fasciola gigantica intermedi-ate forms from the human endemic area of the Nile Delta, Egypt. Infect Genet Evol. 2008; 8: 51-8.

Marcilla A, Bargues MD, Mas-Coma S. A PCR-RFLP assay for the distinction between Fasciola hepatica and Fasciola gigantica. Mol Cell Probes. 2002; 16:327-33.

Lockyer AE, Olson PD, Littlewood DTJ. Utility of complete large and small subunit rRNA genes in resolving the phylogeny of the Neodermata (Platyhelminthes): implications and a review of the cercomer theory. Biol J Linn Soc. 2003; 78: 155-71.

Tkach VV, Littlewood DT, Olson PD, Kinsella JM, Swiderski Z. Molecular phylogenetic analysis of the Microphalloidea Ward, 1901 (Trematoda: Digenea). Syst Parasitol. 2003; 56(1): 1-15.

Wilson WD, Johnson PT, Sutherland DR, Moné H, Loker ES. A molecular phylogenetic study of the genus Ribeiroia (Digenea): trematodes known to cause limb malformations in amphibians. Para-sitol. 2005; 91(5): 1040-5. 17. Choudhury A, Rosas Valdez R, Johnson RC, Hoffmann B, De León GP. The phylogenetic position of allocreadiidae (Trematoda: Digenea) from partial sequences of the 18S and 28S ribo-somal RNA genes. Parasitol. 2007; 93(1): 192-6.

Yamaguti S. Systema helminthum: The digenetic trematodes of vertebrates. Vol. I. New York, USA: Interscience; 1958.

Sahba GH, Arfaa F, Farahmandian I, Jalali H. Animal fascioliasis in Khuzestan, southwestern Iran. J Parasitol. 1972; 58: 712-6.

Soulsby EJL. Helminths, Arthropods and Proto-zoa of domesticated animals. 7th ed. London, UK: Baillière Tindall; 1982.

Salahimoghaddam A. Epidemiology of human fascioliasis in Iran. Journal of Kerman University of Medical Sciences. 2009; 16(4): 385-98. (in Per-sian with English abstract)

Sambrook J, Russell DW. Molecular cloning: a laboratory manual. 3rd ed. New York, USA: Cold Spring Harbor Laboratory Press; 2002.

Walker SM, Prodöhl P, Hoey EM, Fairweather I, Hanna RE, Brennan G, Trudgett A. Substantial genetic divergence between morphologically in-distinguishable populations of Fasciola suggests the possibility of cryptic speciation. Int J Parasitol. 2012; 42(13-14): 1193-9.

Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0., Molecular Biology and Evolution. 2007; 24: 1596-9.

Dosay-Akbuluta M, Trudgettb A, Stanhope M. Understanding genetic diversity of the liver fluke Fasciola hepatica. Z Naturforsch. 2005; 60: 774-8.

Valero MA, Panova M, Mas-Coma S. Phenotypic analysis of adults and eggs of Fasciola hepatica by

computer image analysis system. J Helminthol. 2005; 79: 217-25.

Itagaki T, Sakaguchi K, Terasaki K, Sasaki O, Yoshihara S, Van Dung T. Occurrence of sper-mic diploid and aspermic triploid forms of Fasciola in Vietnam and their molecular characterization based on nuclear and mitochondrial DNA. Para-sitol Int. 2009; 58: 81-5.

Moghaddam AS, Massoud J, Mahmoodi M, Mahvi AH, Periago MV, Artigas P, Fuentes MV, Bargues MD, Mas-Coma S. Human and animal fascioliasis in Mazandaran province, northern Iran. Parasitol Res. 2004; 94: 61-9.

Ashrafi K, Valero MA, Panova M, Periago MV, Massoud J, Mas-Coma S. Phenotypic analysis of adults of Fasciola hepatica, Fasciola gigantica and in-termediate forms from the endemic region of Gi-lan, Iran. Parasitol Int. 2006; 55: 249-60.

Varma AK. On Fasciola indica n. sp. with some observations on F. hepatica and F. gigantica. J Hel-minthol. 1953; 27: 185-98.

Chu JK, Kim YK. Taxonomical study on the Fasciolidae in Korea. Parasitol. 1967; 5: 139-46.

Choe SE, Nguyen TT, Kang TG, Kweon CH, Kang SW. Genetic analysis of Fasciola isolates from cattle in Korea based on second internal transcribed spacer (ITS-2) sequence of nuclear ri-bosomal DNA. Parasitol Res. 2011; 109 (3): 833-9.

Oshima T, Akahane H, Shimazu T. Patterns of the variation of the common liver fluke (Fasciola sp.) in Japan. I. Variations in the sizes and shapes of the worms and eggs. Jpn J Parasitol. 1968; 17: 97-105. (in Japanese with English abstract)

Kimura S, Shimizu A, Kawano J. Morphological observation on liver fluke detected from naturally infected carabaos in the Philippines. Sci Rept Fac Agr Kobe Univ. 1984; 16: 353-7.

Mas-Coma S, Bargues MD. Human liver flukes: a review. Res Rev Parasitol. 1997; 57:145-218.

Vilas R, Vázquez-Prieto S, Paniagua E. Con-trasting patterns of population genetic structure of Fasciola hepatica from cattle and sheep: implica-tions for the evolution of anthelmintic resistance. Infect Genet Evol. 2012; 12(1): 45-52.

Moazeni M, Sharifiyazdi H, Izadpanah A. Char-acterization of Fasciola hepatica genotypes from cat-tle and sheep in Iran using cytochrome C oxidase gene (CO1). Parasitol Res. 2012; 110(6): 2379-84.

Rokni MB, Mirhendi H, Behnia M, Fasihi Harandi M, Jalalizand N. Molecular charac-terization of Fasciola hepatica isolates by RAPD-PCR and Ribosomal ITS1 sequencing. Iranian Red Crescent Medical Journal. 2010a; 12(1): 27-32.

Rokni MB, Mirhendi H, Mizani A, Mohebali M, Sharbatkhori M, Kia EB, Abdoli H, Izadi S. Iden-tification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCR restriction en-zyme method. Exp Parasitol. 2010b; 124: 209-13.

Shahbazi A, Akbarimoghaddam M, Izadi S, Gha-zanchaii A, Jalali N, Bazmani A. Identification and Genetic Variation of Fasciola Species from Tabriz, North-Western Iran. Iran J Parasitol. 2011; 6(3): 52-9.

Mahami-Oskouei M, Dalimi A, Forouzandeh-Moghadam M, Rokni MB. Molecular identifica-tion and differentiation of Fasciola isolates using PCR-RFLP method based on Internal Tran-scribed Spacer (ITS1, 5.8S rDNA, ITS2). Iranian J Parasitol. 2011; 6(3): 35-42.

Tkach VV, Littlewood DT, Olson PD, Kinsella JM, Swiderski Z. Molecular phylogenetic analysis of the Microphalloidea Ward, 1901 (Trematoda: Digenea). Syst Parasitol. 2003; 56(1): 1-15.

Wilson WD, Johnson PT, Sutherland DR, Moné H, Loker ES. A molecular phylogenetic study of the genus Ribeiroia (Digenea): trematodes known to cause limb malformations in amphibians. Para-sitol. 2005; 91(5): 1040-5.

Ashrafi K, Massoud J, Holakouei Nainei K, Jo-Afshani MA, Mahmoodi M, Ebedati N. Nuclear

ribosomal DNA ITS-2 sequence characterization of Fasciola hepatica and Galba truncatula. Iran J Publ Health. 2007; 36: 42-9

Ghavami MB, Rahimi P, Haniloo A, Mo-savinasab SN. Genotypic and phenotypic analysis of Fasciola isolates. Iran J Parasitol. 2009; 4(3): 61-70.

Amor N, Halajian A, Farjallah S, Merella P, Said K, Slimane BB. Molecular characterization of Fasciola spp. from the endemic area of northern Iran based on nuclear ribosomal DNA sequences. Exp Para. 2011; 128: 196-204.

Li QY, Dong SJ, Zhang WY, Lin RQ, Wang CR, Qian DX, Lun ZR, Song HQ, Zhu XQ. Se-quence-related amplified polymorphism, an effec-tive molecular approach for studying genetic vari-ation in Fasciola spp. of human and animal health significance. Electrophoresis. 2009; 30(2): 403-9.

Chaichanasak P, Ichikawa M, Sobhon P, Itagaki T. Identification of Fasciola flukes in Thailand based on their spermatogenesis and nuclear ribo-somal DNA, and their intraspecific relationships based on mitochondrial DNA. Parasitol Int. 2012; 61(4): 545-9.

Itagaki T, Tsutsumi K, Ito K, Tsutsumi Y. Taxo-nomic status of the Japanese triploid forms of Fasciola: comparison of mitochondrial ND1 and COI sequences with F. hepatica and F. gigantica. J Parasitol. 1998; 84: 445-8.


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