Molecular genotyping of Giardia duodenalis in children from Behbahan, southwestern Iran

  • Raziyeh Kasaei
  • David Carmena
  • Ali Jelowdar
  • Molouk Beiromvand
Original Paper


Giardia duodenalis is an intestinal flagellated protozoan that infects humans and several animal species. Giardiasis causing more than 200 million symptomatic infections globally is one of the most common causes of diarrhea in developing countries. Based on molecular studies mainly targeting the small-subunit (SSU) rRNA gene locus of the parasite, eight assemblages (A to H) have been identified in humans and other animal species. The aim of the current study was to evaluate the frequency and molecular diversity of G. duodenalis in children from rural and urban day care centers from Behbahan, southwestern Iran. This cross-sectional study was based on a concentration method for the microscopic detection of G. duodenalis in stool samples of 450 children, aged 1–7 years, in Behbahan, southwestern Iran. The survey was conducted from December 2015 to May 2016. PCR methods targeting the SSU rRNA and triose phosphate isomerase (TPI) genes of G. duodenalis were used for the identification and genotyping of the parasite isolates. Based on sucrose flotation and microscopy techniques, 2.7% (12/450) of children were infected with G. duodenalis, of which six (50.0%) were males and the other six (50.0%) were females. Overall, 91.7% (11/12) of the infections were detected in children from rural areas. The SSU rRNA and TPI genes were amplified successfully in nine and eight, respectively, of the Giardia-positive samples at microscopy. Among the eight TPI sequences, assemblage A, sub-assemblage AII, was identified in five of the isolates. The sequences of the three remaining samples were untypable. Although no significantly statistical difference between genotype and clinical symptoms was found, five out of the eight isolates identified as assemblage A were obtained in asymptomatic children. Giardia duodenalis infections were more prevalent in children from rural day care schools, and the predominant assemblage was A, sub-assemblage AII. The higher prevalence of giardiasis in rural areas might be related to differences in personal hygiene habits, parents’ education level, source of drinking water, and inadequate hygienic toilet facilities in rural areas.


G. duodenalis Assemblage Genotyping Triose phosphate isomerase Children; Iran 



The authors are grateful to day care schoolchildren, parents, and their staffs who participated in the study.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

436_2018_5826_MOESM1_ESM.docx (31 kb)
ESM 1 (DOCX 31 kb)


  1. Arani AS, Alaghehbandan R, Akhlaghi L, Shahi M, Lari AR (2008) Prevalence of intestinal parasites in a population in south of Tehran, Iran. Rev Inst Med Trop Sao Paulo 50:145–149CrossRefPubMedGoogle Scholar
  2. Aydin AF, Besirbellioglu BA, Avci IY, Tanyuksel M, Araz E, Pahsa A (2004) Classification of Giardia duodenalis parasites in Turkey into groups A and B using restriction fragment length polymorphism. Diagn Microbiol Infect Dis 50:147–151. CrossRefPubMedGoogle Scholar
  3. Babaei Z et al (2008) Molecular characterization of the Iranian isolates of Giardia lamblia: application of the glutamate dehydrogenase gene. Iran J Public Health 37:75–82Google Scholar
  4. Baldursson S, Karanis P (2011) Waterborne transmission of protozoan parasites: review of worldwide outbreaks—an update 2004–2010. Water Res 45:6603–6614. CrossRefPubMedGoogle Scholar
  5. Beiromvand M, Mirrezaie E, Mirzavand S (2017) Foodborne giardiasis: is there any relationship between food handlers and transmission of Giardia duodenalis? Infect Disord Drug Targets 17:72–76. CrossRefPubMedGoogle Scholar
  6. Cacciò SM, Ryan U (2008) Molecular epidemiology of giardiasis. Mol Biochem Parasitol 160:75–80. CrossRefPubMedGoogle Scholar
  7. Caccio SM, Lalle M, Svard SG (2017) Host specificity in the Giardia duodenalis species complex. Infect Genet Evol.
  8. Cardona GA, Carabin H, Goñi P, Arriola L, Robinson G, Fernández-Crespo JC, Clavel A, Chalmers RM, Carmena D (2011) Identification and molecular characterization of Cryptosporidium and Giardia in children and cattle populations from the province of Alava, North of Spain. Sci Total Environ 412-413:101–108. CrossRefPubMedGoogle Scholar
  9. Census (2011) Census of the Islamic Republic of Iran, Statistical Centre of Iran. http://www.amarorgir/
  10. Daryani A, Hosseini-Teshnizi S, Hosseini SA, Ahmadpour E, Sarvi S, Amouei A, Mizani A, Gholami S, Sharif M (2017) Intestinal parasitic infections in Iranian preschool and school children: a systematic review and meta-analysis. Acta Trop 169:69–83. CrossRefPubMedGoogle Scholar
  11. de Lucio A, Amor-Aramendía A, Bailo B, Saugar JM, Anegagrie M, Arroyo A, López-Quintana B, Zewdie D, Ayehubizu Z, Yizengaw E, Abera B, Yimer M, Mulu W, Hailu T, Herrador Z, Fuentes I, Carmena D (2016) Prevalence and genetic diversity of Giardia duodenalis and Cryptosporidium spp. among school children in a rural area of the Amhara Region, North-West Ethiopia. PLoS One 11:e0159992. CrossRefPubMedPubMedCentralGoogle Scholar
  12. Faraji R, Ahmadian F, Javadi GR, Barshahi PM (2015) Prevalence of giardiasis among children in childcare centers in Kermanshah, Iran. Int J Res Med Sci 3:1717–1720. CrossRefGoogle Scholar
  13. Feng Y, Xiao L (2011) Zoonotic potential and molecular epidemiology of Giardia species and giardiasis. Clin Microbiol Rev 24:110–140. CrossRefPubMedPubMedCentralGoogle Scholar
  14. Ghafari R, Rafiei A, Tavalla M (2012) Prevalence of intestinal parasites among children referred to Abozar Hospital in Ahvaz. Jundishapur Sci Med 13:627–633 (Persian)Google Scholar
  15. Gillhuber J, Pallant L, Ash A, Thompson RC, Pfister K, Scheuerle MC (2013) Molecular identification of zoonotic and livestock-specific Giardia-species in faecal samples of calves in Southern Germany. Parasit Vectors 6:346. CrossRefPubMedPubMedCentralGoogle Scholar
  16. Goncalves AL, Belizario TL, Pimentel Jde B, Penatti MP, Pedroso Rdos S (2011) Prevalence of intestinal parasites in preschool children in the region of Uberlandia, State of Minas Gerais, Brazil. Rev Soc Bras Med Trop 44:191–193CrossRefPubMedGoogle Scholar
  17. Haque R, Roy S, Kabir M, Stroup SE, Mondal D, Houpt ER (2005) Giardia assemblage A infection and diarrhea in Bangladesh. J Infect Dis 192:2171–2173. CrossRefPubMedGoogle Scholar
  18. Hatam-Nahavandi K, Mohebali M, Mahvi AH, Keshavarz H, Mirjalali H, Rezaei S, Meamar AR, Rezaeian M (2017) Subtype analysis of Giardia duodenalis isolates from municipal and domestic raw wastewaters in Iran. Environ Sci Pollut Res Int 24:12740–12747. CrossRefPubMedGoogle Scholar
  19. Hazrati Tappeh K, Manafi G, Asgharzadeh M, Manafi F (2014) Incidence of Giardia lamblia subspecies by PCR-RFLP in stool specimens of hospitalized children at Urmia Mutahhari Hospital, West Azerbaijan Province, Iran. Iran J Parasitol 9:541–547PubMedPubMedCentralGoogle Scholar
  20. Hijjawi N, Yang R, Mukbel R, Yassin Y, Mharib T, Ryan U (2016) First genetic characterisation of Giardia in human isolates from Jordan. Parasitol Res 115:3723–3729. CrossRefPubMedGoogle Scholar
  21. Homan WL, Mank TG (2001) Human giardiasis: genotype linked differences in clinical symptomatology. Int J Parasitol 31:822–826CrossRefPubMedGoogle Scholar
  22. Hooshyar H, Ghafarinasab S, Arbabi M, Delavari M, Rasti S (2017) Genetic variation of Giardia lamblia isolates from food-handlers in Kashan, Central Iran. Iran J Parasitol 12:83–89PubMedPubMedCentralGoogle Scholar
  23. Kotloff KL, Nataro JP, Blackwelder WC, Nasrin D, Farag TH, Panchalingam S, Wu Y, Sow SO, Sur D, Breiman RF, Faruque ASG, Zaidi AKM, Saha D, Alonso PL, Tamboura B, Sanogo D, Onwuchekwa U, Manna B, Ramamurthy T, Kanungo S, Ochieng JB, Omore R, Oundo JO, Hossain A, Das SK, Ahmed S, Qureshi S, Quadri F, Adegbola RA, Antonio M, Hossain MJ, Akinsola A, Mandomando I, Nhampossa T, Acácio S, Biswas K, O'Reilly CE, Mintz ED, Berkeley LY, Muhsen K, Sommerfelt H, Robins-Browne RM, Levine MM (2013) Burden and aetiology of diarrhoeal disease in infants and young children in developing countries (the Global Enteric Multicenter Study, GEMS): a prospective, case-control study. Lancet 382:209–222. CrossRefPubMedGoogle Scholar
  24. Lagapa JT et al (2009) Monitoring of environmental contamination by Echinococcus multilocularis in an urban fringe forest park in Hokkaido, Japan. Environ Health Prev Med 14:299–303. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Mateo M, Mateo M, Montoya A, Bailo B, Saugar JM, Aguilera M, Fuentes I, Carmena D (2014) Detection and molecular characterization of Giardia duodenalis in children attending day care centers in Majadahonda, Madrid, Central Spain. Medicine (Baltimore) 93:e75. CrossRefGoogle Scholar
  26. Nasiri Goorabi L, Pirestani M, Sadraei J (2017) Genotyping of Giardia duodenalis by β-giardin gene in asymptomatic patients. J Mazandaran Univ Med Sci 27:27–34 (Persian)Google Scholar
  27. Nxasana N, Baba K, Bhat V, Vasaikar S (2013) Prevalence of intestinal parasites in primary school children of Mthatha, Eastern Cape province, South Africa. Ann Med Health Sci Res 3:511–516. CrossRefPubMedPubMedCentralGoogle Scholar
  28. Rafiei A, Roointan ES, Samarbafzadeh AR, Shayesteh AA, Shamsizadeh A, Pourmahdi Borujeni M (2013) Investigation of possible correlation between Giardia duodenalis genotypes and clinical symptoms in southwest of Iran. Iran J Parasitol 8:389–395PubMedPubMedCentralGoogle Scholar
  29. Rafiei A, Rahdar M, Nourozi RV (2014) Isolation and identification of parasitic protozoa in sampled water from the southwest of Iran. Jundishapur J Health Sci 6:e23462. CrossRefGoogle Scholar
  30. Ramirez JD et al (2015) Molecular diagnosis and genotype analysis of Giardia duodenalis in asymptomatic children from a rural area in Central Colombia. Infect Genet Evol 32:208–213. CrossRefPubMedGoogle Scholar
  31. Rayani M, Zasmy Unyah N, Hatam G (2014) Molecular identification of Giardia duodenalis isolates from Fars Province, Iran. Iran J Parasitol 9:70–78PubMedPubMedCentralGoogle Scholar
  32. Read C, Walters J, Robertson ID, Thompson RC (2002) Correlation between genotype of Giardia duodenalis and diarrhoea. Int J Parasitol 32:229–231CrossRefPubMedGoogle Scholar
  33. Robertson LJ, Hanevik K, Escobedo AA, Morch K, Langeland N (2010) Giardiasis—why do the symptoms sometimes never stop? Trends Parasitol 26:75–82. CrossRefPubMedGoogle Scholar
  34. Sahagun J et al (2008) Correlation between the presence of symptoms and the Giardia duodenalis genotype. Eur J Clin Microbiol Infect Dis 27:81–83. CrossRefPubMedGoogle Scholar
  35. Saki J, Amraee D (2017) Prevalence of intestinal parasites among the rural primary school students in the west of Ahvaz county, Iran, 2015. Jentashapir J Health Res 8:e40326Google Scholar
  36. Santos CK et al (2012) Epidemiological, parasitological and molecular aspects of Giardia duodenalis infection in children attending public daycare centers in southeastern Brazil. Trans R Soc Trop Med Hyg 106:473–479. CrossRefPubMedGoogle Scholar
  37. Sarkari B, Ashrafmansori A, Hatam G, Motazedian M, Asgari Q, Mohammadpour I (2012) Genotyping of Giardia lamblia isolates from human in southern Iran. Trop Biomed 29:366–371PubMedGoogle Scholar
  38. Savioli L, Smith H, Thompson A (2006) Giardia and Cryptosporidium join the ‘neglected diseases initiative’. Trends Parasitol 22:203–208. CrossRefPubMedGoogle Scholar
  39. Sprong H, Caccio SM, van der Giessen JW (2009) Identification of zoonotic genotypes of Giardia duodenalis. PLoS Negl Trop Dis 3:e558. CrossRefPubMedPubMedCentralGoogle Scholar
  40. Squire SA, Ryan U (2017) Cryptosporidium and Giardia in Africa: current and future challenges. Parasit Vectors 10:195. CrossRefPubMedPubMedCentralGoogle Scholar
  41. Sulaiman IM, Fayer R, Bern C, Gilman RH, Trout JM, Schantz PM, Das P, Lal AA, Xiao L (2003) Triosephosphate isomerase gene characterization and potential zoonotic transmission of Giardia duodenalis. Emerg Infect Dis 9:1444–1452. CrossRefPubMedPubMedCentralGoogle Scholar
  42. Thomas Iv LJ, Zweig AP, Tosh AK (2014) An adolescent with chronic giardiasis mimicking anorexia nervosa. Int J Adolesc Med Health 26:293–295. CrossRefPubMedGoogle Scholar
  43. Thompson RC, Ash A (2016) Molecular epidemiology of Giardia and Cryptosporidium infections. Infect Genet Evol 40:315–323. CrossRefPubMedGoogle Scholar
  44. Wielinga C, Thompson RCA, Monis P, Ryan U (2015) Identification of polymorphic genes for use in assemblage B genotyping assays through comparative genomics of multiple assemblage B Giardia duodenalis isolates. Mol Biochem Parasitol 201:1–4. CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Raziyeh Kasaei
    • 1
    • 2
  • David Carmena
    • 3
  • Ali Jelowdar
    • 1
    • 2
  • Molouk Beiromvand
    • 1
    • 2
  1. 1.Infectious and Tropical Disease Research Center, Health Research InstituteAhvaz Jundishapur University of Medical SciencesAhvazIran
  2. 2.Department of Parasitology, School of MedicineAhvaz Jundishapur University of Medical SciencesAhvazIran
  3. 3.Parasitology Reference and Research Laboratory, National Centre for MicrobiologyCarlos III Health InstituteMadridSpain

Personalised recommendations