Molecular seasonal, age and gender distributions of Cryptosporidium in diarrhoeic Egyptians: distinct endemicity

  • A. A. El-Badry
  • A. S. A. Al-Antably
  • M. A. Hassan
  • N. A. Hanafy
  • E. Y. Abu-Sarea
Original Article

Abstract

Cryptosporidiosis is a worldwide gastrointestinal disease caused by the protozoan Cryptosporidium parasite. It has a broad range of seasonal and age-related prevalence. We aimed to study the molecular prevalence and seasonality of Cryptosporidium over a period of 1 year in a cohort of Egyptian diarrhoeic patients. Stool samples were collected from 865 diarrhoeic patients attending outpatient clinics of Cairo University hospitals, from all age groups over a 12-month period, examined microscopically for faecal Cryptosporidium oocysts by the acid-fast staining method and for copro-DNA detection using nested polymerase chain reaction (nPCR) assays. PCR-positive samples were characterised molecularly by nPCR-restriction fragment length polymorphism (RFLP) to determine Cryptosporidium genotypes. Cryptosporidium copro-DNA was detected in 19.5 % of the collected samples throughout the year, with a major peak in summer (August) and a small rise in spring (April). Infection was mainly C. hominis (95.8 %) followed by C. parvum (3.0 %), affecting all age groups, with predominance in the pre-school age group, and decrease with age. There were statistically significant associations between the detection of Cryptosporidium and season, diarrhoea, patient age and drinking water, while gender, contact with animals and presence of mucus in stool showed no association. Cryptosporidium in diarrhoeic Egyptians was of distinct endemicity, with the bi-model mostly influenced by population dynamics, with a clear high prevalence in pre-school children and predominating anthroponotic (C. hominis) transmission throughout the year. The obtained results highlight Cryptosporidium as a water contaminant and an important cause of health problems in Egypt, necessitating further studies of the risk factors.

References

  1. 1.
    Smith HV, Cacciò SM, Cook N, Nichols RA, Tait A (2007) Cryptosporidium and Giardia as foodborne zoonoses. Vet Parasitol 149:29–40CrossRefPubMedGoogle Scholar
  2. 2.
    Neglected Diseases Initiative of the World Health Organization. Home page at: http://www.who.int/neglected_diseases/en/
  3. 3.
    Sulaiman IM, Hira PR, Zhou L, Al-Ali FM, Al-Shelahi FA, Shweiki HM, Iqbal J, Khalid N, Xiao L (2005) Unique endemicity of cryptosporidiosis in children in Kuwait. J Clin Microbiol 43(6):2805–2809PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Youssef FG, Adib I, Riddle MS, Schlett CD (2008) A review of cryptosporidiosis in Egypt. J Egypt Soc Parasitol 38:9–28PubMedGoogle Scholar
  5. 5.
    Xiao L (2010) Molecular epidemiology of cryptosporidiosis: an update. Exp Parasitol 124(1):80–89CrossRefPubMedGoogle Scholar
  6. 6.
    Ghallab MMI, Abdel-Aziz IZ, Shoeib EY, El-Badry AA (2014) Laboratory utility of coproscopy, copro immunoassays and copro nPCR assay targeting Hsp90 gene for detection of Cryptosporidium in children, Cairo, Egypt. J Parasit Dis 8(2):1–5Google Scholar
  7. 7.
    Peng MM, Xiao L, Freeman AR, Arrowood MJ, Escalante AA, Weltman AC, Ong CS, Mac Kenzie WR, Lal AA, Beard CB (1997) Genetic polymorphism among Cryptosporidium parvum isolates: evidence of two distinct human transmission cycles. Emerg Infect Dis 3:567–573PubMedCentralCrossRefPubMedGoogle Scholar
  8. 8.
    Lal A, Hales S, French N, Baker MG (2012) Seasonality in human zoonotic enteric diseases: a systematic review. PLoS One 7(4):31883CrossRefGoogle Scholar
  9. 9.
    Pedraza-Díaz S, Amar C, Nichols GL, McLauchlin J (2001) Nested polymerase chain reaction for amplification of the cryptosporidium oocyst wall protein gene. Emerg Infect Dis 7(1):49–56PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Spano F, Putignani L, McLauchlin J, Casemore DP, Crisanti A (1997) 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 150:209–217CrossRefPubMedGoogle Scholar
  11. 11.
    Abd El Kader NM, Blanco MA, Ali-Tammam M, Abd El Ghaffar Ael R, Osman A, El Sheikh N, Rubio JM, de Fuentes I (2011) Detection of Cryptosporidium parvum and Cryptosporidium hominis in human patients in Cairo, Egypt. Parasitol Res 110(1):161–166CrossRefPubMedGoogle Scholar
  12. 12.
    El-Settawy MA, Fathy GM (2012) Evaluation and comparison of PCR, coproantigen ELISA and microscopy for diagnosis of Cryptosporidium in human diarrheic specimens. J Am Sci 8(12):1378–1385Google Scholar
  13. 13.
    Fathy MM, Abdelrazek NM, Hassan FA, El-Badry AA (2014) Molecular copro-prevalence of Cryptosporidium in Egyptian children and evaluation of three diagnostic methods. Indian Pediatr 51(9):727–729CrossRefPubMedGoogle Scholar
  14. 14.
    Yoder JS, Beach MJ; Centers for Disease Control and Prevention (CDC) (2007) Giardiasis surveillance—United States, 2003–2005. MMWR Surveill Summ 56(7):11–18PubMedGoogle Scholar
  15. 15.
    Newman RD, Sears CL, Moore SR, Nataro JP, Wuhib T, Agnew DA, Guerrant RL, Lima AAM (1999) Longitudinal study of Cryptosporidium infection in children in Northeastern Brazil. J Infect Dis 180:167–175CrossRefPubMedGoogle Scholar
  16. 16.
    Adamu H, Petros B, Hailu A, Petry F (2010) Molecular characterization of Cryptosporidium isolates from humans in Ethiopia. Acta Trop 115(1–2):77–83CrossRefPubMedGoogle Scholar
  17. 17.
    Natividad FF, Buerano CC, Lago CB, Mapua CA, De Guzman BB, Seraspe EB, Samentar LP, Endo T (2008) Prevalence rates of Giardia and Cryptosporidium among diarrheic patients in the Philippines. Southeast Asian J Trop Med Public Health 39(6):991–999PubMedGoogle Scholar
  18. 18.
    Peng MM, Meshnick SR, Cunliffe NA, Thindwa BD, Hart CA, Broadhead RL, Xiao L (2003) Molecular epidemiology of cryptosporidiosis in children in Malawi. J Eukaryot Microbiol 50:557–559CrossRefPubMedGoogle Scholar
  19. 19.
    Tumwine JK, Kekitiinwa A, Nabukeera N, Akiyoshi DE, Rich SM, Widmer G, Feng X, Tzipori S (2003) Cryptosporidium parvum in children with diarrhea in Mulago Hospital, Kampala, Uganda. Am J Trop Med Hyg 68:710–715PubMedGoogle Scholar
  20. 20.
    Bern C, Ortega Y, Checkley W, Roberts JM, Lescano AG, Cabrera L, Verastegui M, Black RE, Sterling C, Gilman RH (2002) Epidemiologic differences between cyclosporiasis and cryptosporidiosis in Peruvian children. Emerg Infect Dis 8:581–585PubMedCentralCrossRefPubMedGoogle Scholar
  21. 21.
    Learmonth JJ, Ionas G, Ebbett KA, Kwan ES (2004) Genetic characterization and transmission cycles of Cryptosporidium species isolated from humans in New Zealand. Appl Environ Microbiol 70:3973–3978PubMedCentralCrossRefPubMedGoogle Scholar
  22. 22.
    Helmy YA, Krücken J, Nöckler K, von Samson-Himmelstjerna G, Zessin KH (2013) Molecular epidemiology of Cryptosporidium in livestock animals and humans in the Ismailia province of Egypt. Vet Parasitol 193:15–24CrossRefPubMedGoogle Scholar
  23. 23.
    Eida AM, Eida MM, El-Desoky A (2009) Pathological studies of different genotypes of human Cryptosporidium Egyptian isolates in experimentally mice. J Egypt Soc Parasitol 39(3):975–990PubMedGoogle Scholar
  24. 24.
    El-Shazly AM, El-sheikha HM, Soltan DM, Mohammad KA, Morsy TA (2007) Protozoal pollution of surface water sources in Dakahlia Governorate, Egypt. J Egypt Soc Parasitol 37(1):51–64Google Scholar
  25. 25.
    Gatei W, Greensill J, Ashford RW, Cuevas LE, Parry CM, Cunliffe NA, Beeching NJ, Hart CA (2003) Molecular analysis of the 18S rRNA gene of Cryptosporidium parasites from patients with or without human immunodeficiency virus infections living in Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. J Clin Microbiol 41:1458–1462PubMedCentralCrossRefPubMedGoogle Scholar
  26. 26.
    Leav BA, Mackay MR, Anyanwu A, O’Connor RM, Cevallos AM, Kindra G, Rollins NC, Bennish ML, Nelson RG, Ward HD (2002) Analysis of sequence diversity at the highly polymorphic Cpgp40/15 locus among Cryptosporidium isolates from human immunodeficiency virus-infected children in South Africa. Infect Immun 70:3881–3890PubMedCentralCrossRefPubMedGoogle Scholar
  27. 27.
    Xiao L, Fayer R (2008) Molecular characterisation of species and genotypes of Cryptosporidium and Giardia and assessment of zoonotic transmission. Int J Parasitol 38(11):1239–1255CrossRefPubMedGoogle Scholar
  28. 28.
    Al-Brikan FA, Salem HS, Beeching N, Hilal N (2008) Multilocus genetic analysis of Cryptosporidium isolates from Saudi Arabia. J Egypt Soc Parasitol 38:645–658PubMedGoogle Scholar
  29. 29.
    Hijjawi N, Ng J, Yang R, Atoum MF, Ryan U (2010) Identification of rare and novel Cryptosporidium GP60 subtypes in human isolates from Jordan. Exp Parasitol 125:161–164CrossRefPubMedGoogle Scholar
  30. 30.
    Iqbal J, Khalid N, Hira PR (2011) Cryptosporidiosis in Kuwaiti children: association of clinical characteristics with Cryptosporidium species and subtypes. J Med Microbiol 60(Pt 5):647–652CrossRefPubMedGoogle Scholar
  31. 31.
    Alyousefi NA, Mahdy MA, Lim YA, Xiao L, Mahmud R (2013) First molecular characterization of Cryptosporidium in Yemen. Parasitology 140(6):729–734CrossRefPubMedGoogle Scholar
  32. 32.
    Alves M, Xiao L, Sulaiman I, Lal AA, Matos O, Antunes F (2003) Subgenotype analysis of Cryptosporidium isolates from humans, cattle, and zoo ruminants in Portugal. J Clin Microbiol 41:2744–2747PubMedCentralCrossRefPubMedGoogle Scholar
  33. 33.
    McLauchlin J, Amar C, Pedraza-Díaz S, Nichols GL (2000) Molecular epidemiological analysis of Cryptosporidium spp. in the United Kingdom: results of genotyping Cryptosporidium spp. in 1,705 fecal samples from humans and 105 fecal samples from livestock animals. J Clin Microbiol 38:3984–3990PubMedCentralPubMedGoogle Scholar
  34. 34.
    Garvey P, McKeown P (2009) Epidemiology of human cryptosporidiosis in Ireland, 2004–2006: analysis of national notification data. Euro Surveill 14(8). pii: 19128Google Scholar
  35. 35.
    Llorente MT, Clavel A, Goñi MP, Varea M, Seral C, Becerril R, Suarez L, Gómez-Lus R (2007) Genetic characterization of Cryptosporidium species from humans in Spain. Parasitol Int 56(3):201–205CrossRefPubMedGoogle Scholar
  36. 36.
    Cama VA, Bern C, Roberts J, Cabrera L, Sterling CR, Ortega Y, Gilman RH, Xiao L (2008) Cryptosporidium species and subtypes and clinical manifestations in children, Peru. Emerg Infect Dis 14:1567–1574PubMedCentralCrossRefPubMedGoogle Scholar
  37. 37.
    El-Sherbini GT, Abosdera MM (2013) Risk factors associated with intestinal parasitic infections among children. J Egypt Soc Parasitol 43(1):287–294CrossRefPubMedGoogle Scholar
  38. 38.
    Abdel-Messih IA, Wierzba TF, Abu-Elyazeed R, Ibrahim AF, Ahmed SF, Kamal K, Sanders J, Frenck R (2005) Diarrhea associated with Cryptosporidium parvum among young children of the Nile River Delta in Egypt. J Trop Pediatr 51:154–159CrossRefPubMedGoogle Scholar
  39. 39.
    Hlavsa MC, Watson JC, Beach MJ (2004) Cryptosporidiosis surveillance—United States, 1999–2002. In: Surveillance Summaries. MMWR 54(1):1–8Google Scholar
  40. 40.
    ANOFEL Cryptosporidium National Network (2010) Laboratory-based surveillance for Cryptosporidium in France, 2006–2009. Euro Surveill 15(33):19642Google Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.Medical Parasitology Department, Kasr Al Ainy School of MedicineCairo UniversityEl-Manial, CairoEgypt
  2. 2.Department of Medical Parasitology, Faculty of MedicineBeni Suef UniversityBeni SuefEgypt

Personalised recommendations