Articles

Molecular Identification of Giardia duodenalis Isolates from Fars Province, Iran

Abstract

Background: Giardia duodenalis is one of the most common human intestinal pro-tozoan parasites worldwide and is endemic throughout the world with a vast range of mammalian hosts. The present study aimed to identify the prevalence of G. duo-denalis isolates and determine the most common of its assemblages in the patients referring to health centers and hospitals in Fars province, Iran that will be subjected to further molecular investigation.

Methods: We collected 1000 human fecal samples from health centers and hospi-tals in Shiraz, Iran in a one year period from September 2009 to August 2010. Mi-croscopic examination for the presence of G. duodenalis cysts and trophozoites was performed by direct wet mount before and after the concentration techniques. Ex-traction of DNA was performed by Phenol-Chloroform-Isoamylalcohol (PCI). G. duodenalis-positive specimens were analyzed by PCR. A fragment of the SSU-rDNA (292 bp) gene was amplified by PCR using the forward primer RH11 and the re-verse primer RH4. Genotyping was performed using sequence analysis of G. duode-nalis glutamate dehydrogenase gene using primers GDHeF, GDHiF, and GDHiR.

Results: The prevalence of Giardia infection was 10.7% (107/1000) examined based on microscopic examination. PCR identified 80% (40/50) of the samples as positive for G. duodenalis based on SSU-rDNA amplification on sucrose gradient samples. Besides, genotyping results indicated 32 isolates (80%) as assemblage AII and 8 isolates (20%) as assemblage BIII and BIV based on the DNA sequence analysis of the glutamate dehydrogenase locus of G. duodenalis.

Conclusion: The findings of this study emphasize that Iran (Fars Province) is a favorable area for giardiasis with an anthroponotic infection route.

Johnston AR, Gillespie, TR, Rwego, IB, McLachlan, TLT, Kent AD, Goldberg TL. Molecular epidemiology of cross-species Giardia duodenalis transmission in western Uganda. PLoS Negl Trop Dis. 2010; 4(5), 683.

Adam RD. The biology of Giardia spp. Microbiological Reviews. 1991; 55(4), 706-732.

Thompson, R., Monis, P. Variation in Giardia: implications for taxonomy and epidemiology. Adv Parasitol. 2004; 58, 69-137.

Volotao A, Costa-Macedo L, Haddad F, Brandao A, Peralta J, Fernandes O. Genotyping of Giardia duodenalis from human and animal samples from Brazil using [beta]-giardin gene: A phylogenetic analysis. Acta Trop. 2007; 102(1), 10-19.

Kosuwin R, Putaporntip C, Pattanawong U, Jongwutiwes S. Clonal diversity in Giardia duodenalis isolates from Thailand: Evidences for intragenic recombination and purifying selection at the beta giardin locus. Gene. 2010; 449(1), 1-8.

Yang R, Lee J, Ng J, Ryan U. High prevalence Giardia duodenalis assemblage B and potentially zoonotic subtypes in sporadic human cases in Western Australia. Int J Parasitol. 2010; 40(3), 293-297.

Gelanew T, Lalle M, Hailu A, Pozio E, Caccio SM. Molecular characterization of human isolates of Giardia duodenalis from Ethiopia. Acta Trop. 2007; 102(2), 92-99.

Feng Y, Xiao L. Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev. 2011; 24(1), 110-140.

Berrilli F, D'Alfonso R, Giangaspero A, Marangi M, Brandonisio O, Kaboré Y, et al. Giardia duodenalis genotypes and Cryptosporidium species in humans and domestic animals in Cote d’Ivoire: occurrence and evidence for environmental contamination. Trans R Soc Trop Med Hyg. 2012; 106(3), 191-195.

Read C, Walters J, Robertson ID, Thompson RC. Correlation between genotype of Giardia duodenalis and diarrhoea. Int J Parasitol. 2002; 32(2), 229-31.

Tungtrongchitr A, Sookrung N, Indrawattana N, Kwangsi S, Ongrotchanakun J, Chaicumpa W. Giardia intestinalis in Thailand: identification of genotypes. J Health Popul Nutr. 2010; 28(1), 42-52.

Arani AS, Alaghehbandan R, Akhlaghi L, Shahi M, Lari AR. Prevalence of intestinal parasites in a population in south of Tehran, Iran. Rev Inst Med Trop Sao Paulo. 2008; 50(3), 145-149.

Mohammed Mahdy A, Surin J, Wan K, Mohd-Adnan A, Al-Mekhlafi M, Lim Y. Giardia intestinalis genotypes: Risk factors and correlation with clinical symptoms. Acta Trop. 2009; 112(1), 67-70.

Caccio SM, Ryan U. Molecular epidemiology of giardiasis. Mol Biochem Parasitol. 2008; 160(2), 75-80.

Wang R, Zhang X, Zhu H, Zhang L, Feng Y, Jian F, et al. Genetic characterizations of Cryptosporidium spp. and Giardia duodenalis in humans in Henan, China. Exp Parasitol. 2011; 127(1), 42-45.

Bertrand I, Albertini L, Schwartzbrod J. Comparison of two target genes for detection and genotyping of Giardia lamblia in human feces by PCR and PCR-restriction fragment length polymorphism. J Clin Microbiol. 2005; 43(12), 5940-5944.

Sousa M, Morais J, Machado J, Poiares Da Silva J. Genotyping of Giardia lamblia Human Isolates from Portugal by PCR RFLP and Sequencing. J Eukaryot Microbiol. 2006; 53(S1), S174-S176. 18. Thompson RCA. Towards a better unders-tanding of host specificity and the transmission of Giardia: the impact of molecular epidemiology. In: Olson, B.E., Olson, M.E. and Wallis, P.M., (eds.) Giardia the cosmopolitan parasite. CABI Publishing, Wallingford, 2002. p. 55-69.

Sarkari B, Ashrafmansori A, Hatam GR, Mota-zedian MH, Asgari Q, Mohamma-dpour I. Genotyping of Giardia lamblia isolates from human in southern Iran. Trop Biomed. 2012; 29 (3), 366-371.

Shoorijeh SJ, Sadjjadi SM, Asheri A, Eraghi K. Giardia Spp. And Sarcocystis Spp. Status In Pet Dogs Of Shiraz, Southern Part Of Iran. Trop Bi-omed. 2008; 25 (2), 154-159.

Haghighi A, Khorashad AS, Mojarad EN, Kazemi B, Nejad MR, Rasti S. Frequency of enteric protozoan parasites among patients with gastrointestinal complaints in medical centers of Zahedan, Iran. Trans R Soc Trop Med Hyg. 2009; 103(5), 452-454.

Taherkhani H, Shariati S, Abdolahi N, Rosha-ndel G. Clinical manifestations of giardiasis in Iran. J Clin Diagn Res. 2009; 3, 1416-1418.

Hopkins RM, Meloni BP, Groth DM, Wether-all JD, Reynoldson JA, Thompson RC. Ribo-somal RNA sequencing reveals differences between the genotypes of Giardia isolates recovered from humans and dogs living in the same locality. J Parasitol. 1997; 44-51.

Santín M, Trout JM, Vecino JA, Dubey JP, Fayer R. Cryptosporidium, Giardia and Enter-ocytozoon bieneusi in cats from Bogota (Colombia) and genotyping of isolates. Vet Parasitol. 2006; 141(3), 334-339.

Read CM, Monis PT, Thompson RC. Discrimination of all genotypes of Giardia duodenalis at the glutamate dehydrogenase locus using PCR-RFLP. Infect Genet Evol. 2004; 4(2), 125-130.

Nantavisai K, Mungthin M, Tan-ariya P, Ran-gsin R, Naaglor T, Leelayoova S. Evaluation of the sensitivities of DNA extraction and PCR methods for detection of Giardia duodenalis in stool specimens. J Clin Microbiol. 2007; 45(2), 581-583.

Orlandi PA, Lampel KA. Extraction-free, filter-based template preparation for rapid and sensitive PCR detection of pathogenic parasitic protozoa. J Clin Microbiol. 2000; 38(6), 2271-2277.

Traub RJ, Monis PT, Robertson I, Irwin P, Mencke N, Thompson RC. Epidemiological and molecular evidence supports the zoonotic transmission of Giardia among humans and dogs living in the same community. Parasitology. 2004; 128(03), 253-262.

Wielinga CM, Thompson RC. Comparative evaluation of Giardia duodenalis sequence data. Parasitology. 2007; 134(12), 1795-821.

Ey PL, Mansouri M, Kulda J, Nohýnková E, Monis PT, Andrews RH, et al. Genetic Analysis of Giardia from Hoofed Farm Animals Reveals Artiodactyl-Specific and Potentially Zoonotic Genotypes. J Eukaryot Microbiol. 1997; 44(6), 626-635. 31. Monis PT, Mayrhofer G, Andrews RH, Homan WL, Limper L, Ey PL. Molecular genetic analysis of Giardia intestinalis isolates at the glutamate dehydrogenase locus. Parasitology. 1996; 112, 1-12.

Monis PT, Andrews RH, Mayrhofer G, Mackrill J, Kulda J, Isaac-Renton JL, et al. Novel lineages of Giardia intestinalis identified by genetic analysis of organisms isolated from dogs in Australia. Parasitology. 1998; 116(1), 7-19.

Bajer A. Cryptosporidium and Giardia spp. Infect-ions in humans, animals and the environment in Poland. Parasitol Res. 2008; 104(1), 1-17.

Boontanom P, Siripattanapipong S, Mungthin M, Tan-ariya P, Leelayoova S. Improved sensitivity of PCR amplification of glutamate dehydrogenase gene for detection and genotyping of Giardia duodenalis in stool specimen. Southeast Asian J Trop Med Public Health. 2010; 41(2), 280-284

Sayyari AA, Imanzadeh F, Bagheri Yazdi SA, Karami H, Yaghoobi M. Prevalence of intestinal parasitic infections in the Islamic Republic of Iran. East Mediterr Health J. 2005; 11(3). 377-83.

Kia E, Hosseini M, Nilforoushan M, Meamar A, Rezaeian M. Study of intestinal protozoan parasites in rural inhabitants of Mazandaran province, Northern Iran. Iran J Parasitol. . 2008; 3(1), 21-25.

Souza SL, Gennari SM, Richtzenhain LJ, Pena HF, Funada MR, Cortez A, et al. Molecular identification of Giardia duodenalis isolates from humans, dogs, cats and cattle from the state of Sao Paulo, Brazil, by sequence analysis of fragments of glutamate dehydrogenase (gdh) coding gene. Vet Parasitol. 2007; 149(3-4), 258-264.

Babaei Z, Oormazdi H, Akhlaghi L, Rezaie S, Razmjou E, Soltani-Arabshahi S, et al. Molecular characterization of the Iranian isolates of Giardia lamblia: application of the glutamate dehydro-genase gene. Iranian J Publ Health. 2008; 37(2), 75-82.

Caccio SM, De Giacomo M, Pozio E. Sequence analysis of the beta-giardin gene and develo-pment of a polymerase chain reaction-restriction fragment length polymorphism assay to geno-type Giardia duodenalis cysts from human faecal samples. Int J Parasitol. 2002; 32(8), 1023-1030.

Winkworth CL, Learmonth JJ, Matthaei CD, Townsend CR. Molecular characterization of Giardia isolates from calves and humans in a region in which dairy farming has recently intensified. Appl Environ Microbiol. 2008; 74(16), 5100-5105.

Traub RJ., Inpankaew T, Reid SA, Sutthikornchai C, Sukthana Y, Robertson ID, et al. Transmission cycles of Giardia duodenalis in dogs and humans in Temple communities in Bangkok-a critical evalu-ation of its prevalence using three diagnostic tests in the field in the absence of a gold standard. Acta Trop. 2009; 111(2), 125-132.

Haque R, Roy S, Kabir M, Stroup SE, Mondal D, Houpt ER. Giardia assemblage A infection and diarrhea in Bangladesh. J Infect Dis . 2005; 192(12), 2171-2173.

Geurden T, Levecke B, Cacció SM, Visser A, De Groote G, Casaert S, et al. Multilocus genotyping of Cryptosporidium and Giardia in non-outbreak related cases of diarrhoea in human patients in Belgium. Parasitology. 2009; 136(10), 1161-1168.

Breathnach AS, McHugh TD, Butcher PD. Prevalence and clinical correlations of genetic subtypes of Giardia lamblia in an urban setting. Epidemiol Infect. 2010; 138(10), 1459-1467.

Lebbad M, Ankarklev J, Tellez A, Leiv-a B, Andersson JO, Svard S. Dominance of Giardia assemblage B in Leon, Nicaragua. Acta Trop. 2008; 106(1), 44-53.

Yason JA, Rivera WL. Genotyping of Giardia duodenalis isolates among residents of slum area in Manila, Philippines. Parasitol Res. 2007; 101(3), 681-687.

Cooper MA, Sterling CR, Gilman RH, Cama V, Ortega Y, Adam RD. Molecular analysis of household transmission of Giardia lamblia in a region of high endemicity in Peru. J Infect Dis . 2010; 202(11), 1713-1721.

Files
IssueVol 9 No 1 (2014) QRcode
SectionArticles
Keywords
Giardia duodenalis Iran Microscopic Molecular characterization Prevalence S SU-rDNA

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
How to Cite
1.
Rayani M, Zasmy UNYAH N, Hatam G. Molecular Identification of Giardia duodenalis Isolates from Fars Province, Iran. Iran J Parasitol. 1;9(1):70-78.