PCR-RFLP Analysis of 28 SrDNA for Specification of Fasciola gigantica (Cobbold, 1855) in the Infected Lymnaea auricularia (Linnaeus, 1785) Snails from Northwestern Iran.
AbstractBackground: Fasciolosis in livestock is a crucial concern in the globe, mainly due to its impact on human health. The aim of this study was to determine the rate of infection with Fasciola gigantica (Cobbold, 1855) larvae in the field-collected snails of Lymnaea auricularia (Linnaeus, 1785) from northwestern Iran using a molecular approach.Methods: A total of 6,759 pond snails were collected from 28 freshwater bodies in West Azarbaijan. PCR was performed to amplify a 618-bp fragment of the nuclear 28 SrRNA gene of Fasciola. The PCR products were digested by AvaII restriction enzyme to create restriction fragment length polymorphism (RFLP) patterns specific for the detection of F. gigantica.Results: Of the total collected snails 496 (7.34 %) were L. auricularia, among which 4.64% (23 out of 496) were infected with a Fasciola species according to the PCR analysis. Only 2.22% (11 out of 496) of the infected snails were from the mountainous areas. The highest Fasciola infection rate recorded in the snails of a single site was 1.81% (9 out of 496 snails). Based on the RFLP pattern, F. gigantica accounted for 2.42% of the infection rates in the study sites.Conclusion: Application of PCR-RFLP was proven to be a useful approach for valid and robust detection of the infection with F. gigantica in its intermediate host snails. These findings may therefore be applicable for establishment of the controlprograms against dissemination of the infection in different regions.
Yakhchali M, Ghobadi K. A survey on helminths infection of liver and economic loss in slaughtered sheep of Urmia slaughterhouse, Iran. Iran Vet J. 2005; 11: 60-5. [Persian].
Bargues MD, Vigo M, Horak P, Dvorak J, Patzner RA, Pointier JP, Jackiewicz M, Meier- Brook C, Mas-Coma S. European Lymnaeidae (Mollusca: Gastropoda), intermediate hosts of trematodiases, based on nuclear ribosomal DNA ITS-2 sequences. Infect Genet Evol. 2001; 1: 85-107.
Hurtrez-Boussès S, Meunier C, Durand P, Renaud F. Dynamics of host parasite interactions: the example of population biology of the liver fluke (Fasciola hepatica). Microb Infect. 2001; 3: 841-9.
Lotfy WM, Brant SV, De Jong RJ, Le TH, Demiaszkiewicz A, Rajapakse RPVJ, Perera VBVP, Laursen JR, Loker ES. Evolutionary origins, diversification, and biogeography of liver flukes (Digenea, Fasciolidae). Am J Trop Med Hyg. 2008; 79(2):248–55.
Mera y Sierra RMY, Artigas P, Cuervo P, Deis E, Sidoti L, Mas-Coma S, Bargues MD. Fascioliasis transmission by Lymnaea neotropica confirmed by nuclear rDNA and mtDNA sequencing in Argentina. Vet Parasitol. 2009; 166: 73–9.
Mansoorian AB. A practical guidline for identification of Iranian freshwater snails. Iran J Public Health. 1986; 15(1-2): 41-53.
Imani-Baran A, Yakhchali M, Malekzadeh-Viayeh R. A study on geographical distribution and diversity of Lymnaeidae snails in West Azerbaijan Province, Iran. Veterinary Journal(Pajouhesh and Sazandegi). 2011a; 82: 53-63.[Persian]
Ghobadi K, Yakhchali M. Survey of helminthes infection rate of liver and economical losses in Urmia slaughterhouse, Iran. Journal of Faculty of Veterinary Medicine, Tehran University. 2003; 58(1): 19-20. [Persian]
Caron Y, Rondelaud D, Losson B. The detection and quantification of a digenian infection in the snail host with special emphasis on Fasciola sp. Parasitol Res. 2008; 103: 735-44.
McGarry JW, Ortiz PL, Hodgkinson JE, Goreish I, Williams DJL. PCR-based differentiation of Fasciola species (Trematoda: Fasciolidae), using primers based on RAPD derived sequences. Ann Trop Med Parasitol. 2007;, 101(5): 415 - 21.
Ashrafi K, Massoud J, Holakouei K, Mahmoodi M, Joafshani MA, Valero MA, Fuentes MV, Khoubbane M, Artigas P, Bargues MD, Mas-Coma S. Evidence Suggesting that Fasciola gigantica might be the most prevalent causal agent of Fascioliasis in Northern Iran. Iran J Public Health. 2004; 33(4): 31-7.
Athari A, Gohar-Dehi SH, Rostami M, Jalilian MD. Determination of definitive and intermediate hosts of cercarial dematitis-producing agents in Northern Iran. Arc Iran Med. 2006; 9: 11-5.
Farahnak A, Essalat MA. A study on cercarial dermatitis in Khoozestan province, southwestern Iran. BMC Public Health. 2003; 3: 35-8.
Imani-Baran A, Yakhchali M, Malekzadeh- Viayeh R, Farhangpajuh F. Prevalence of cercariae infection in Lymnaea auricularia (Linnaeus,1758) in North West of Iran. Vet Res Forum. 2011b; 2(2): 121-7.
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.
Mahami-Oskouei M, Dalimi A, Forouzandeh- Moghadam M, Rokni MB. Molecular identification and differentiation of Fasciola isolates using PCR- RFLP method based on internal transcribed spacer (ITS1, 5.8S rDNA, ITS2). Iran J Parasitol. 2011; 6(3): 35-42.
Shahbazi A, Akbarimoghaddam M, Izadi S, Ghazanchaii 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.
Pfleger V. A field guide in colour to molluscs. Prague, Czech Republic: Aventinum Nakladatelstyi, S.T.O., Polygrafia; 1999. p. 28-9.
Faltynkova A, Nasincova V, Kablaskova L. Larval trematodes (digenea) of Planorbid snails (Gastropoda: Pulmonata) in Central Europe: a survey of species and key to their identification. Syst Parasitol. 2008; 69:155-78.
Sambrook J, Russell DW. Molecular cloning: A laboratory manual. 3rd ed., New York, USA: Cold Spring Harbor Laboratory Press; 2002.
Ashrafi K, Massoud J, Holakouie Naieni K, Jo- Afshani MA, Mahmoodi M, Ebadati N, Rezvani SM, Artigas P, Mas-Coma S, Bargues MD. Nuclear ribosomal DNA ITS-2 sequence characterization of Fasciola hepatica and Galba truncatula. Iran J Public Health. 2007; 36: 42–9.
Karimi A. Genetic diagnosis of Fasciola species based on 18S ribosomal DNA sequences. J Biol Sci. 2008; 8: 1166–73.
Huang WY, He B, Wang CR, Zhu XQ. Characterisation of Fasciola species from Mainland China by ITS-2 ribosomal DNA sequence. Vet Parasitol. 2004; 120: 75–83.
Rokni MB, Mirhendi H., Mizani A, Mohebali M, Sharbatkhori M, Kia EB, Abdoli H, Izad S. Identification and differentiation of Fasciola hepatica and Fasciola gigantica using a simple PCRrestriction enzyme method. Exp Parasitol. 2010; 124: 209–13.
Saki J, Khademvatan S, Yousefi E. Molecular identification of animal Fasciola isolates in southwest of Iran. Aust J Basic Appl Sci. 2011; 5(11): 1878-83.
Massoud J, Sadjadi S. Susceptibility of different species of Lymnaea snails to miracidia of Fasciola gigantica and F. hepatica in Iran. J Helminthol. 1980; 54(3): 201-2.
Sharif M, Daryani A, Karimi SA. A faunistic survey of cercariae isolated from lymnaeid snails in central areas Mazandaran, Iran. Pak J Biol Sci. 2010; 13(4): 158-63.
Fuentes MV, Valero MA, Bargues MD, Esteban JG, Angles R, Mas-Coma S. Analysis of climatic data and forecast indices for human fascioliasis at very high altitude. Ann Trop Med Parasitol 1999; 93: 835-50.
Boray JC. The potential impact of exotic Lymnaea spp. on fascioliasis in Australasia. Vet Parasitol. 1978; 4: 127-41.
Al-Kubaisee RY, Altaif KI. Comparative studies of sheep and buffalo isolates of Fasciola gigantica in the intermediate host Lymnaea auricularia. Res Vet Sci. 1989; 47: 273-74.
Mas-Coma S, Valero MA, Bargues MD. Climate change effects on trematodiases, with emphasis on zoonotic fascioliasis and schistosomiasis. Vet Parasitol. 2009; 163: 264-80.
Salahimoghaddam A. Epidemiology of human fasciolosis in Iran. Journal of Kerman University of Medical Sciences. 2009; 16(4): 385-98.[Persian]
Hamburger J, Hoffman O, Kariuki HC, Muchiri EM, Ouma JH, Koech DK, Sturrock RF, King CH. Large-scale, polymerase chain reaction-based surveillance of Schistosoma haematobium DNA in snails from transmission sites in coastal Kenya: a new tool for studying the dynamics of snail infection. Am J Trop Med Hyg. 2004; 71: 765-73.