Original Article

Assessment of Giardia and Cryptosporidium Assemblages/ Species and Their Viability in Potable Tap Water in Beni-Suef, Egypt Using Nested PCR/RFLP and Staining


Background: The protozoan Giardia and Cryptosporidium are responsible for most water-borne diseases all over the world. The extent and number of outbreaks of waterborne diseases suggests a significant risk of their potential transmission via drinking water. This study aimed to document the prevalence and viability of Giardia and Cryptosporidium (oo) cysts in tap water samples in Beni-Suef Governorate, Egypt and to detect the predominant Giardia and Cryptosporidium assemblages/species using nested PCR/ Restriction Fragment Length Polymorphism (RFLP) confirmed by further sequencing of positive samples.

Methods: A total of 80 tap water samples were collected throughout a year from four big centers and filtered using the membrane filtration method. Samples were stained by Lugol’s iodine, Modified Zeihl-Neelsen (MZN) (to detect prevalence) and trypan blue stain (to detect viability). Nested PCR-RFLP and sequencing were used for molecular characterizations and genotyping of the detected Giardia and Cryptosporidium.

Results: Giardia and Cryptosporidium DNA was detected in 20 (25%) and 29 (36.3%) samples respectively, with predominance of Giardia assemblage B (85%) and C. hominis (75.9%). The prevalence and viability of both parasites (oo) cysts showed seasonality which peaked in summer and were greater in Beba center and in rural areas.

Conclusion: To our knowledge, no studies have been done in these areas before. The anthroponotic transmission has an important role in giardiasis and crypto­sporidiosis epidemiology in this studied area.

Céline M, Aurélien D, Sylvie G, Christelle-Laurent M. Monitoring of Cryptosporidium and Giardia river contamination in Paris area. Water Res. 2009; 43(1):211-17.

Baldursson S, Karanis P. Waterborne transmission of protozoan parasites: review of worldwide outbreaks - an update 2004-2010. Water Res. 2011; 45: 6603-6614

Adam RD. Biology of Giardia lamblia.Clin Microbiol Rev. 2001; 14, 447–475.

Fayer R. Cryptosporidium a water-borne zoonotic parasite. Vet Parasitol. 2004; 126: 37-56.

DuPont HL, Chappell CL, Sterling CR, Okhuysen PC, Rose JB, Jakubowski W. The infectivity of Cryptosporidium parvum in healthy volunteers. New Eng J Med. 1995; 332, 855–859.

Mac Kenzie WR, Hoxie NJ, Proctor ME, Gradus MS, Blair KA, Peterson DE, Kazmierczak JJ, Addiss DJ, Fox KR, Rose JB, Davis JP. A massive outbreak in Milwaukee of Cryptosporidium infection transmitted through the Public Water Supply. N Engl J Med. 1994; 331: 161-167.

Hunter PR, Thompson RC. The zoonotic transmission of Giardia and Cryptosporidium.Int J Parasitol. 2005; 35: 1181-1190.

Karanis P, Kourenti C, Smith H. Waterborne outbreaks of protozoan parasites: A worldwide review of outbreaks and lessons learnt. J Water Health. 2007; 5(1):1-38.

Khalifa AM, Ibrahim IR, Said DE, Aleem EA, Nabil RA. Cryptosporidium and Giardia in Water in Alexandria: Detection and Evaluation of Viability by Flow Cytometry and Different Stains. PUJ. 2011; 4(2): 155-164.

El Shazly AM, Elsheikha HM, Soltan DM, Mohammad KA, Morsy TA. Protozoal pollution of surface water sources in Dakahlia governorate. J Egypt Soc Parasitol. 2007; 37(1):51- 64.

Dechesne M, Soyeux EJ. Assessment of source water pathogen contamination. Water Health. 2007; 5 (1):39-50.

Ryan U, Hijjawi N. New developments in Cryptosporidium research. Int J Parasitol. 2015; 45: 367-373.

Rossle N, Latif B. Cryptosporidiosis as threatening health problem: a review. Asian Pac J Trop Biomed. 2013; 3: 916-924.

Thompson RCA, Monis PT. Taxonomy of Giardia species. Luján HD, Svard S, eds. Giardia: a model organism. Springer; 2011; 3-15.

Durigan M, Abreu AG, Zuchi MI, Franco RMB, de Souza AP. Genetic diversity of Giardia duodenalis: Multilocus genotyping reveals zoonotic potential between clinical and environmental sources in a metropolitan region of Brazil. PLoS ONE. 2014; 9: 115489.

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

El-Kowrany SI, El- Zamarany EA, El-Nouby KA, El-Mehy DA, Abo Ali EA, Othman AA, Salah W, El-Ebiary AA. Water pollution in the Middle Nile Delta, Egypt: An environmental study.J of Adv Res. 2016; 7 (5):781-794.

Sakran TF, El-Shahawy GA, Shalaby MA, Sabry HY, Matooq PM, Elmallah AM. Detection rates of waterborne protozoa in water sources from Fayoum Governorate. PUJ, 2017; 10 (1): 30-38.

Giacometti A, Cirioni O, Barchiesi F, Ancarani F, Scalise G. Activity of nitazoxanide alone and in combination with azithromycin and rifabutin against Cryptosporidium parvum in cell culture. J Antimicrob Chemother. 2000; 5(5):453-6.

Spano F, Putignani L, McLauchlin J, Casemore DP, Crisanti A. PCR-RFLP analysis of the Cryptosporidium oocyst wall protein (COWP) gene discriminates between C. wrairi and C. parvum, and between C. parvum isolates of human and animal origin. FEMS Microbiol Lett. 1997; 150:209–217.

Pedraza-Díaz S, Amar C, Nichols GL, McLauchlin J. Nested polymerase chain reaction for amplification of the Cryptosporidium oocyst wall protein gene. Emerg Infect Dis. 2001; 7(1):49–56.

Caccio SM, de Giacomo M, Pozio E. Sequence analysis of the β-giardin gene and development of a PCR-RFLP assay to genotype Giardia duodenalis cysts from human faecal samples. Int J Parasitol. 2002; 32:1023–1030

Lallea M, Pozioa E, Capellib G, Bruschic F, Crottid D, Caccio SM. Genetic heterogeneity at the b-giardin locus among human and animal isolates of Giardia duodenalis and identification of potentially zoonotic subgenotypes. Int J Parasitol. 2005; 35:207–213

Almeida A, Moreira MJ, Soares S, Delgado MD, Figueiredo J, Silva E, Castro A , Da Cosa JMC. Presence of Cryptosporidium spp. and Giardia duodenalis in Drinking Water Samples in the North of Portugal. Korean J Parasitol. 2010; 48 (1): 43-48.

Ali MA, Al-Herrawy AZ, El-Hawaary SE. Detection of enteric viruses, Giardia and Cryptosporidium in two different types of drinking water treatment facilities. Water Res. 2004; 38(18): 3931-9.

Khalifa AM, Yacout MAF, Sadek AA. Genetical and electron microscopical studies on Cryptosporidia. J Egypt Soc Parasitol. 2001; 31(3):799-814.

Sayed FG, Hamza AI, Galal LA, Sayed DM ,Gaber M. Detection of Cryptosporidium parvum oocysts contaminating hospitals drinking water supply using different techniques during winter/summer season. Glo Adv Res J Microbiol. 2016; 5(6):068-079.

Khalifa RMA, Ahmad AK, Abdel-Hafeez EH, Mosllem FA. Present Status Of Protozoan Pathogens Causing Water Borne Disease In Northen Part Of El-Minia Governorate, Egypt. J Egypt Soc Parasitol. 2014; 44(3), 559–566.

Carmena D, Aguinagalde X, Zigorraga C, Fernandez-Crespo JC, Ocio JA. Presence of Giardia cysts and Cryptosporidium oocysts in drinking water supplies in northern Spain. Journal of Applied Microbiology. 2007; 102 619–629.

Nichols RAB, Campbell BM, Smith HV. Identification of Cryptosporidium spp. oocysts in United Kingdom noncarbonated natural mineral waters and drinking waters by using a modified nested PCR-restriction fragment length polymorphism assay. Appl Environ Microbiol. 2003; 69:4183–9.

Hashimoto A, Kunikane S, Hirata T. Prevalence of Cryptosporidium oocysts and Giardia cysts in the drinking water supply in Japan. Water Res. 2002; 36:519–26.

Koompapong K, Sukthana K. Seasonal variation and potential sources of Cryptosporidium contamination in surface waters of Chao Phraya River and Bang Pu Nature Reserve Pier, Thailand. South Asian J Trop Med Pub Heal. 2012; 43(4):832–840.

El-Badry AA, Al-Antably AS, Hassan MA, Hanafy NA, Abu-Sarea EY. Molecular seasonal, age and gender distributions of Cryptosporidium in diarrhoeic Egyptians: distinct endemicity.Eur J Clin Microbiol Infect Dis. 2015; 34(12):2447-2453

Ismail MA, El-Akkad DM, Rizk EM, El-Askary HM, El-Badry AA. Molecular seasonality of Giardia lamblia in a cohort of Egyptian children: a circannual pattern. Parasitol Res. 2016; 115 (11): 4221-4227.

Lal A, Baker MG, Hales S, French NP. Potential effects of global environmental changes on cryptosporidiosis and giardiasis transmission. Trends Parasitol. 2013; 29(2):83–90.

Cama VA, Bern C, Roberts J, Cabrera L, Sterling CR, Ortega Y, Gilman RH, Xiao L. Cryptosporidium species and subtypes and clinical manifestations in children, Peru. Emerg Infect Dis. 2008; 14: 1567–1574.

Ryan U, Caccio S. Zoonotic potential of Giardia. Int J Parasitol 2013; 43,943–956.

Mossallam SF. Detection of some intestinal protozoa in commercial fresh juices. J Egypt Soc Parasitol. 2010; 40(1):135-49.

El-Sherbini GT, Abosdera MM. Risk factors associated with intestinal parasitic infections among children. J Egypt Soc Parasitol. 2013; 43(1):287–294.

IssueVol 14 No 3 (2019) QRcode
SectionOriginal Article(s)
DOI https://doi.org/10.18502/ijpa.v14i3.1475
Cryptosporidium spp Tap water Egypt Giardia

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How to Cite
HAMDY D, El-BADRY A, ABD EL WAHAB W. Assessment of Giardia and Cryptosporidium Assemblages/ Species and Their Viability in Potable Tap Water in Beni-Suef, Egypt Using Nested PCR/RFLP and Staining. Iran J Parasitol. 2019;14(3):368-378.