Abstract
Cryptosporidium species are enteric apicomplexan parasites associated with diarrhoeal disease in humans and animals globally. Waterborne outbreaks resulting from contamination with the infective oocysts are common worldwide. Updated reports on waterborne protozoal infections are needed to identify emerging pathogens and susceptible populations. Therefore, this study aimed to assess the current profile of Cryptosporidium contamination of various water sources in Sharqia Governorate, Northeastern Egypt. For this purpose, eighty samples were collected from five different water types (canal, tap, tank, filtered, and groundwater), distributed in four major cities (El-Hessenia, Fakous, Zagazig, and Belbies) in Sharqia Governorate. All water samples were examined using conventional microscopy, ELISA, and real-time PCR (RT-PCR) techniques. Based on microscopic analysis, the Cryptosporidium protozoan was identified in 25% of the tested water samples. The RT-PCR assay has allowed for the quantification of Cryptosporidium oocysts in different types of water. Canal water exhibited the highest Cryptosporidium contamination levels (mean = 85.15 oocysts/L), followed by water tanks (mean = 12.031 oocysts/L). The study also provided a comparative evaluation of ELISA and RT-PCR for the diagnosis of Cryptosporidium infection. RT-PCR performed better than ELISA in terms of analytical accuracy (97.50% vs. 86.25%) and specificity (100% vs. 83.33%). However, ELISA showed a higher sensitivity (95.00% vs. 90.00%) for Cryptosporidium recovery. Our findings could serve as a platform for further investigations into the potential risks associated with water contamination in Sharqia Governorate.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data and materials used to support the findings of this study are included within this article.
References
Akuffo I, Cobbina SJ, Alhassan EH, Nkoom M (2013) Assessment of the quality of water before and after storage in the Nyankpala community of the Tolon-kumbungu district, Ghana. Int J Sci Technol Res 2:221–227
Aldom JE, Chagla AH (1995) Recovery of Cryptosporidium oocysts from water by a membrane filter dissolution method. Lett Appl Microbiol 20(3):186–187. https://doi.org/10.1111/j.1472-765X.1995.tb00423.x
Ali MA, Al-Herrawy AZ, El-Hawaary SE (2004) Detection of enteric viruses, Giardia and Cryptosporidium in two different types of drinking water treatment facilities. Water Res 38(18):3931–3939. https://doi.org/10.1016/j.watres.2004.06.014
Anceno AJ, Katayama H, Houpt ER, Chavalitshewinkoon-Petmitr P, Chuluun B, Shipin OV (2007) IMS-free DNA extraction for the PCR-based quantification of Cryptosporidium parvum and Giardia lamblia in surface and waste water. Int J Environ Health Res 17(4):297–310. https://doi.org/10.1080/09603120701372573
Castro-Hermida JA, González-Warleta M, Mezo M (2015) Cryptosporidium spp. and Giardia duodenalis as pathogenic contaminants of water in Galicia, Spain: the need for safe drinking water. Int J Hyg Environ Health 218(1):132–138. https://doi.org/10.1016/j.ijheh.2014.09.001
Chalmers RM, Campbell BM, Crouch N, Charlett A, Davies AP (2011) Comparison of diagnostic sensitivity and specificity of seven Cryptosporidium assays used in the UK. J Med Microbiol 60(Pt 11):1598–1604. https://doi.org/10.1099/jmm.0.034181-0
Chapman PA, Rush BA, McLauchlin J (1990) An enzyme immunoassay for detecting Cryptosporidium in faecal and environmental samples. J Med Microbiol 32(4):233–237. https://doi.org/10.1099/00222615-32-4-233
Chique C, Hynds PD, Andrade L, Burke L, Morris D, Ryan MP, O’Dwyer J (2020) Cryptosporidium spp. in groundwater supplies intended for human consumption - a descriptive review of global prevalence, risk factors and knowledge gaps. Water Res 1(176):115726. https://doi.org/10.1016/j.watres.2020.115726
Elfadaly HA, Hassanain NA, Hassanain MA, Barakat AM, Shaapan RM (2018) Evaluation of primitive ground water supplies as a risk factor for the development of major waterborne zoonosis in Egyptian children living in rural areas. J Infect Public Health 11(2):203–208. https://doi.org/10.1016/j.jiph.2017.07.025
El-Kowrany SI, El-Zamarany EA, El-Nouby KA, El-Mehy DA, Ali EAA, Othman AA, Salah W, El-Ebiary AA (2016) Water pollution in the middle Nile Delta, Egypt: an environmental study. J Adv Res 7(5):781–794. https://doi.org/10.1016/j.jare.2015.11.005
Elmehy DA, Ismail HIH, Alfattah AA, Elnouby KA, Hazza SM, El-Badry AA (2021) Flow cytometric and molecular analysis of possible protozoal contamination of drinking water in Tanta, Egypt. J Egypt Soc Parasitol 51(1):127–138. https://doi.org/10.21608/JESP.2021.165953
Elshazly AM, Elsheikha HM, Soltan DM, Mohammad KA, Morsy TA (2007) Protozoal pollution of surface water sources in Dakahlia Governorate. Egypt J Egypt Soc Parasitol 37(1):51–64
El-Zanfaly HT (2015) Water quality and health in Egyptian rural areas. J Environ Prot Sustain Dev 1(4):203–210
Feng Y, Zhao X, Chen J, Jin W, Zhou X, Li N, Wang L, Xiao L (2011) Occurrence, source, and human infection potential of cryptosporidium and Giardia spp. in source and tap water in Shanghai, China. Appl Environ Microbiol 77(11):3609–3616. https://doi.org/10.1128/AEM.00146-11
Gad MA, Saleh F, Morsy EA, Marouf MA, Al-Herrawy AZ (2019) Use of microscopic and molecular techniques to assess removal of parasitic protozoa via conventional and compact drinking water treatment processes. Egypt J Aquat Biol Fish 23(5):327–339. https://doi.org/10.21608/EJABF.2019.67228
Godiwala NT, Vandewalle A, Ward HD, Leav BA (2006) Quantification of in vitro and in vivo Cryptosporidium parvum infection by using real-time PCR. Appl Environ Microbiol 72(6):4484–4488. https://doi.org/10.1128/AEM.00189-06
Graham JP, VanDerslice J (2007) The effectiveness of large household water storage tanks for protecting the quality of drinking water. J Water Health 5(2):307–313. https://doi.org/10.2166/wh.2007.011b
Guy RA, Payment P, Krull UJ, Horgen PA (2003) Real-time PCR for quantification of Giardia and Cryptosporidium in environmental water samples and sewage. Appl Environ Microbiol 69(9):5178–5185. https://doi.org/10.1128/AEM.69.9.5178-5185.2003
Hadfield SJ, Robinson G, Elwin K, Chalmers RM (2011) Detection and differentiation of Cryptosporidium spp. in human clinical samples by use of real-time PCR. J Clin Microbiol 49(3):918–924. https://doi.org/10.1128/jcm.01733-10
Hadi AM (2014) Isolation and identification of Cryptosporidium spp. by reverse osmosis system of tap water in Baghdad. Baghdad Sci J 11:894–899. https://doi.org/10.21123/bsj.2014.11.2.894-899
Hamdy D, El-Badry A, Abd El Wahab W (2019) 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 14(3):368–378. https://doi.org/10.18502/ijpa.v14i3.1475
Helmi K, Skraber S, Burnet JB, Leblanc L, Hoffmann L, Cauchie HM (2011) Two-year monitoring of Cryptosporidium parvum and Giardia lamblia occurrence in a recreational and drinking water reservoir using standard microscopic and molecular biology techniques. Environ Monit Assess 179(1–4):163–175. https://doi.org/10.1007/s10661-010-1726-7
Hijjawi N, Estcourt A, Yang R, Monis P, Ryan U (2010) Complete development and multiplication of Cryptosporidium hominis in cell-free culture. Vet Parasitol 169(1–2):29–36. https://doi.org/10.1016/j.vetpar.2009.12.021
Hsu BM, Wu NM, Jang HD, Shih FC, Wan MT, Kung CM (2005) Using the flow cytometry to quantify the Giardia cysts and Cryptosporidium oocysts in water samples. Environ Monit Assess 104(1–3):155–162. https://doi.org/10.1007/s10661-005-1608-6
Jiang J, Alderisio KA, Singh A, Xiao L (2005) Development of procedures for direct extraction of Cryptosporidium DNA from water concentrates and for relief of PCR inhibitors. Appl Environ Microbiol 71(3):1135–1141. https://doi.org/10.1128/AEM.71.3.1135-1141.2005
Keeley A, Faulkner BR (2008) Influence of land use and watershed characteristics on protozoa contamination in a potential drinking water resources reservoir. Water Res 42(10–11):2803–2813. https://doi.org/10.1016/j.watres.2008.02.028
Khalifa RM, Ahmad AK, Abdel-Hafeez EH, Mosllem FA (2014) Present status of protozoan pathogens causing water-borne disease in northern part of El-Minia Governorate, Egypt. J Egypt Soc Parasitol 44(3):559–566. https://doi.org/10.12816/0007860
Khalil IA, Troeger C, Rao PC, Blacker BF, Brown A, Brewer TG, Colombara DV, De Hostos EL, Engmann C, Guerrant RL, Haque R, Houpt ER, Kang G, Korpe PS, Kotloff KL, Lima AAM, Petri WA Jr, Platts-Mills JA, Shoultz DA, Forouzanfar MH, Hay SI, Reiner RC Jr, Mokdad AH (2018) Morbidity, mortality, and long-term consequences associated with diarrhoea from Cryptosporidium infection in children younger than 5 years: a meta-analyses study. Lancet Glob Health 6(7):e758–e768. https://doi.org/10.1016/S2214-109X(18)30283-3
King P, Tyler KM, Hunter PR (2019) Anthroponotic transmission of Cryptosporidium parvum predominates in countries with poorer sanitation: a systematic review and meta-analysis. Parasit Vectors 12(1):16. https://doi.org/10.1186/s13071-018-3263-0
Kumar T, Abd Majid MA, Onichandran S, Jaturas N, Andiappan H, Salibay CC, Tabo HA, Tabo N, Dungca JZ, Tangpong J, Phiriyasamith S, Yuttayong B, Polseela R, Do BN, Sawangjaroen N, Tan TC, Lim YA, Nissapatorn V (2016) Presence of Cryptosporidium parvum and Giardia lamblia in water samples from Southeast Asia: towards an integrated water detection system. Infect Dis Poverty. https://doi.org/10.1186/s40249-016-0095-z
Lindergard G, Wade SE, Schaaf S, Barwick RS, Mohammed HO (2001) Detection of Cryptosporidium oocysts in soil samples by enzyme-linked immunoassay. Vet Parasitol 94(3):163–176. https://doi.org/10.1016/s0304-4017(00)00379-4
Ma JY, Li MY, Qi ZZ, Fu M, Sun TF, Elsheikha HM, Cong W (2022) Waterborne protozoan outbreaks: an update on the global, regional, and national prevalence from 2017 to 2020 and sources of contamination. Sci Total Environ 806(Pt 2):150562. https://doi.org/10.1016/j.scitotenv.2021.150562
Martins FDC, Ladeia WA, Toledo RDS, Garcia JL, Navarro IT, Freire RL (2019) Surveillance of Giardia and Cryptosporidium in sewage from an urban area in Brazil. Rev Bras Parasitol Vet 28(2):291–297. https://doi.org/10.1590/S1984-29612019037
Momčilović S, Cantacessi C, Arsić-Arsenijević V, Otranto D, Tasić-Otašević S (2019) Rapid diagnosis of parasitic diseases: current scenario and future needs. Clin Microbiol Infect 25(3):290–309. https://doi.org/10.1016/j.cmi.2018.04.028
Morris A, Robinson G, Swain MT, Chalmers RM (2019) Direct sequencing of Cryptosporidium in stool samples for public health. Front Public Health 11(7):360. https://doi.org/10.3389/fpubh.2019.00360
Ndao M (2009) Diagnosis of parasitic diseases: old and new approaches. Interdiscip Perspect Infect Dis 2009:278246. https://doi.org/10.1155/2009/278246
Ngowi HA (2020) Prevalence and pattern of waterborne parasitic infections in eastern Africa: a systematic scoping review. Food Waterborne Parasitol 20:e00089. https://doi.org/10.1016/j.fawpar.2020.e00089
Nichols RA, Campbell BM, Smith HV (2003) 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 69(7):4183–4189. https://doi.org/10.1128/AEM.69.7.4183-4189.2003
Nishi L, Baesso ML, Santana RG, Fregadolli P, Falavigna DL, Falavigna-Guilherme AL (2009) Investigation of Cryptosporidium spp. and Giardia spp. in a public water-treatment system. Zoonoses Public Health 56(5):221–228. https://doi.org/10.1111/j.1863-2378.2008.01189.x
Omoruyi BE, Nwodo UU, Udem CS, Okonkwo FO (2014) Comparative diagnostic techniques for cryptosporidium infection. Molecules 19(2):2674–2683. https://doi.org/10.3390/molecules19022674
Pumipuntu N, Piratae S (2018) Cryptosporidiosis: a zoonotic disease concern. Vet World 11(5):681–686. https://doi.org/10.14202/vetworld.2018.681-686
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. https://doi.org/10.5812/jjhs.23462
Ramadan EM, Shalash OS, Fahmy MR, Abdel-Aal GM (2019) Integrated water resource management in Sharkia Governorate, East Nile delta using numerical evaluation of water management strategies. Alex Eng J 58:757–771. https://doi.org/10.1016/j.aej.2019.06.006
Rayan HZ, Eida OM, El-Hamshary EM, Ahmed SA (2009) Detection of human Cryptosporidium species in surface water sources in Ismailia using polymerase chain reaction. PUJ 2(2):119–126
Razakandrainibe R, Mérat C, Kapel N, Sautour M, Guyot K, Gargala G, Ballet JJ, Le Pape P, Dalle F, Favennec L (2021) Multicenter evaluation of an ELISA for the detection of Cryptosporidium spp. antigen in clinical human stool samples. Microorganisms 9(2):209. https://doi.org/10.3390/microorganisms9020209
Razzolini MT, da Silva Santos TF, Bastos VK (2010) Detection of Giardia and Cryptosporidium cysts/oocysts in watersheds and drinking water sources in Brazil urban areas. J Water Health 8(2):399–404. https://doi.org/10.2166/wh.2009.172
Rochelle PA, Marshall MM, Mead JR, Johnson AM, Korich DG, Rosen JS, De Leon R (2002) Comparison of in vitro cell culture and a mouse assay for measuring infectivity of Cryptosporidium parvum. Appl Environ Microbiol 68(8):3809–3817. https://doi.org/10.1128/AEM.68.8.3809-3817.2002
Sakran TF, El-Shahawy GA, Shalaby MA, Sabry HY, Matooq PM, Almahy M, Elmallah AM (2017) Detection rates of waterborne protozoa in water sources from Fayoum Governorate. PUJ 10(1–2):30–38. https://doi.org/10.21608/PUJ.2017.4734
Sánchez C, López MC, Galeano LA, Qvarnstrom Y, Houghton K, Ramírez JD (2018) Molecular detection and genotyping of pathogenic protozoan parasites in raw and treated water samples from southwest Colombia. Parasit Vectors 11(1):563. https://doi.org/10.1186/s13071-018-3147-3
Shafey D, Aboamer MM, Diab KA, Abdallah HM, Yousef MF, Gouda MA (2019) Prevalence of Giardia intestinalis and Cryptosporidium parvum parasites in drinking water in Menoufia Governorate, Egypt. Int J Curr Microbiol App Sci 8(12):2263–2276. https://doi.org/10.20546/ijcmas.2019.812.269
Sim J, Wright CC (2005) The kappa statistic in reliability studies: use, interpretation, and sample size requirements. Phys Ther 85(3):257–268. https://doi.org/10.1093/ptj/85.3.257
Stancari RCA (2013) Sensitivity of ELISA for detecting Cryptosporidium spp. and Giardia spp. in raw waters samples. R Inst Adolfo Lutz 72(3):234–238
Stinear T, Matusan A, Hines K, Sandery M (1996) Detection of a single viable Cryptosporidium parvum oocyst in environmental water concentrates by reverse transcription-PCR. Appl Environ Microbiol 62(9):3385–3390. https://doi.org/10.1128/aem.62.9.3385-3390.1996
Sunaga F, Tsuchiaka S, Kishimoto M, Aoki H, Kakinoki M, Kure K, Okumura H, Okumura M, Okumura A, Nagai M, Omatsu T, Mizutani T (2020) Development of a one-run real-time PCR detection system for pathogens associated with porcine respiratory diseases. J Vet Med Sci 82(2):217–223. https://doi.org/10.1292/jvms.19-0063
Versi E (1992) “Gold standard” is an appropriate term. BMJ 305(6846):187. https://doi.org/10.1136/bmj.305.6846.187-b
Wolyniak EA, Hargreaves BR, Jellison KL (2010) Seasonal retention and release of Cryptosporidium parvum oocysts by environmental biofilms in the laboratory. Appl Environ Microbiol 76(4):1021–1027. https://doi.org/10.1128/AEM.01804-09
Xiao L (2010) Molecular epidemiology of Cryptosporidiosis: an update. Exp Parasitol 124(1):80–89. https://doi.org/10.1016/j.exppara.2009.03.018
Zakai HA, Barnawi HI (2014) Prevalence of Cryptosporidium and Giardia lamblia in water samples from Jeddah and Makkah cities. J Adv Lab Res Biol 5(1):12–17
Cheesbrough M (1991) Techniques used to identify parasite in: Medical Laboratory Manual for Tropical Countries. Britain, Butterworth- Heinemann Ltd., Oxford, UK, pp 178–197
Feiz Hadad MH, Karamkhani A, Haddad RF (2016) Waterborne parasites: a recent status of occurrence, source and human intestinal parasites in sources and tap water; Dehloran, South West, Iran. Allergy Drugs Clin Immunol 118–1121. https://api.semanticscholar.org/CorpusID:212558421
UNICEF/WHO (2011) Drinking Water: Equity, Safety and Sustainability. JMP Thematic Report on Drinking Water
Funding
This research did not receive any specific grants from funding agencies in the public, commercial, or not-for-profit sectors.
Author information
Authors and Affiliations
Contributions
All authors contributed to the study conception and design. The first draft of the manuscript was written by Marwa Omar. Material preparation, data collection and analysis were performed by Marwa Omar and Samia E. Etewa. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors have no relevant financial or non-financial interests to disclose.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Below is the link to the electronic supplementary material.
Rights and permissions
Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.
About this article
Cite this article
Omar, M., Etewa, S.E., Mahmoud, S.A.M. et al. Assessment of the potential occurrence of Cryptosporidium species in various water sources in Sharqia Governorate, Egypt. J Parasit Dis (2024). https://doi.org/10.1007/s12639-024-01675-1
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s12639-024-01675-1