Abstract
Cryptosporidium spp. cause cryptosporidiosis in humans through zoonotic and anthroponotic transmission. Previous studies illustrated the significance of domestic animals as reservoirs of this parasite. Cryptosporidium occurs in Njoro River; a main source of water to humans and animals. However, there is no information on the Cryptosporidium spp. and genotypes circulating in Njoro Sub County. A total of 2174 samples from humans, cattle, chickens, sheep and goats were assessed for the presence of Cryptosporidium spp. Thirty-three positive samples were subsequently successfully sequenced and compared to Cryptosporidium sequences in the GenBank repository using NCBI’s (National Center for Biotechnology Information) online BLAST (Basic Local Alignment Search Tool) algorithmic program. Sequence alignment was done using the Clustal W program and phylogenetic analysis was executed in MEGA X (Molecular Evolutionary Genetics Analysis version X). The prevalence of cryptosporidiosis in Njoro Sub County is 6.99%. Cryptosporidium spp. present in the watershed showed great genetic diversity and nine Cryptosporidium species were recorded: Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium ubiquitum, Cryptosporidium meleagridis, Cryptosporidium andersoni, Cryptosporidium baileyi, Cryptosporidium muris, Cryptosporidium xiaoi and Cryptosporidium viatorum. This is the first study to report the presence of C. viatorum in Kenya. Cattle is the major reservoir of zoonotic Cryptosporidium spp. while goats harbored the lowest number of species. Humans and domestic animals drink the contaminated water from Njoro River, humans are therefore, exposed to a high cryptosporidiosis risk.
Similar content being viewed by others
Data availability
The datasets that support findings of this study have been deposited in a repository, GenBank, under accession numbers MZ435761–MZ435786.
References
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(3):154–159. https://doi.org/10.1093/tropej/fmh105
Adamu H, Petros B, Zhang G, Kassa H, Amer S, Ye J, Feng Y, Xiao L (2014) Distribution and clinical manifestations of Cryptosporidium species and subtypes in HIV/AIDS patients in Ethiopia. PLoS Negl Trop Dis 8(4):e2831. https://doi.org/10.1371/journal.pntd.0002831
Ayinmode AB, Oliveira BCM, Obebe OO, Dada-Adgebola HO, Ayede AI, Widmer G (2018) Genotypic characterization of Cryptosporidium species in humans and peri-domestic animals in Ekiti and Oyo States. Nigeria J Parasitol 104(6):639–644. https://doi.org/10.1645/17-74
Ayinmode AB, Zhang H, Dada-Adegbola HO, Xiao L (2014) Cryptosporidium hominis subtypes and Enterocytozoon bieneusi genotypes in HIV-infected persons in Ibadan Nigeria. Zoonoses Public Hlth 61(4):297–303. https://doi.org/10.1111/zph.12072
Azami M, Moghaddam D, Salehi R, Salehi M (2007) The identification of Cryptosporidium species in Isfahan, Iran by PCR-RFLP analysis of the 18S rRNA gene. Mol Biol 41:851–856. https://doi.org/10.1134/S0026893307050196
Babaei Z, Oormazdi H, Rezaie S, Rezaeian M, Razmjou E (2011) Giardia intestinalis: DNA extraction approaches to improve PCR results. Exp Parasitol 128:159–162. https://doi.org/10.1016/j.exppara.2011.02.001
Berrilli F, D’Alfonso R, Giangaspero A, Marangi M, Brandonisio O, Kaboré Y (2012) Giardia duodenalis genotypes and Cryptosporidium species in humans and domestic animals in Côte d’Ivoire:occurrence and evidence for environmental contamination. Trans R Soc Trop Med Hyg 106(3):191–195. https://doi.org/10.1016/j.trstmh.2011.12.005
Braima K, Zahedi A, Oskam C, Reid S, Pingault N, Xiao L (2019) Retrospective analysis of Cryptosporidium species in Western Australian human populations, (2015–2018), and emergence of the C. hominis IfA12G1R5 subtype. Infect Genet Evol 73:306–313. https://doi.org/10.1016/j.meegid.2019.05.018
Cacciò S, Chalmers R (2016) Human cryptosporidiosis in Europe. Clin Microbiol Infect 22(6):471–480. https://doi.org/10.1016/j.cmi.2016.04.021
Casemore D (1990) Epidemiological aspects of human cryptosporidiosis. Epidemiol Infect 104(1):1–28. https://doi.org/10.1017/s0950268800054480
Casemore DP, Wright SE, Coop RL (1997) Cryptosporidiosis: human and animal epidemiology. In: Fayer R (ed) Cryptosporidium and cryptosporidiosis. CRC Press, Boca Raton, USA, pp 65–92
Castro-Hermida JA, Pors I, Méndez-Hermida F, Ares-Mazás E, Chartier C (2006) Evaluation of two commercial disinfectants on the viability and infectivity of Cryptosporidium parvum oocysts. Vet J 171(2):340–345. https://doi.org/10.1016/j.tvjl.2004.11.003
Cegielski JP, Ortega YR, McKee S, Madden JF, Gaido L, Schwartz DA, Manji K, Jorgensen AF, Miller SE, Pulipaka UP, Msengi AE, Mwakyusa DH, Sterling CR, Reller LB (1999) Cryptosporidium, Enterocytozoon, and Cyclospora infections in pediatric and adult patients with diarrhea in Tanzania. Clin Infect Dis 28(2):314–321. https://doi.org/10.1086/515131
Chalmers RM (2012) Waterborne outbreaks of cryptosporidiosis. Ann 1st Super Sanita 48(4):429–446. https://doi.org/10.4415/ANN_12_04_10
Chappell CL, Okhuysen PC, Langer-Curry RC, Lupo PJ, Widmer G, Tzipori S (2015) Cryptosporidium muris: infectivity and illness in healthy adult volunteers. Am J TropMed Hyg 92(1):50–55. https://doi.org/10.4269/ajtmh.14-0525
Chappell CL, Okhuysen PC, White AC (2003) Cryptosporidium parvum: infectivity, pathogenesis and the host-parasite relationship. Cryptosporidium. https://doi.org/10.1016/b978-044451351-9/50006-9
Chen Y, Zheng W, Zhang N, Gui B, Lv Q, Yan J (2019) Identification of Cryptosporidium viatorum XVa subtype family in two wild rat species in China. J Parasitol Vector Biol 12:502. https://doi.org/10.1186/s13071-019-3763-6
Cordell RL, Addiss DG (1994) Cryptosporidiosis in childcare settings: a review of the literature and recommendations for prevention and control. Pediatr Infect Dis J 13(4):310–317
Current WL (1994) Cryptosporidium parvum: household transmission. Ann Intern Med 120:518–519. https://doi.org/10.7326/0003-4819-120-6-199403150-00012
Current WL, Garcia LS (1991) Cryptosporidiosis. Clin Microbiol Rev 4(3):325–358. https://doi.org/10.1128/CMR.4.3.325
Daugschies A, Bangoura B, Lendner M (2013) Inactivation of exogenous endoparasite stages by chemical disinfectants: current state and perspectives. Parasitol Res 112(3):917–932. https://doi.org/10.1007/s00436-013-3324-4
de Lucio A, Amor-Aramendía A, Bailo B, Saugar JM, Anegagrie M, Arroyo A (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(7):e0159992. https://doi.org/10.1371/journal.pone.0159992
Elwin K, Hadfield SJ, Robinson G, Crouch ND, Chalmers RM (2012) Cryptosporidium viatorum n. sp. (Apicomplexa: Cryptosporidiidae) among travellers returning to Great Britain from the Indian subcontinent 2007–2011. Int J Parasitol 42:675–682. https://doi.org/10.1016/j.ijpara.2012.04.016
Feng Y, Ryan UM, Xiao L (2018) Genetic diversity and population structure of Cryptosporidium. Trends Parasitol 34(11):997–1011. https://doi.org/10.1016/j.pt.2018.07.009
Gonin P, Trudel L (2003) Detection and differentiation of Entamoeba histolytica and Entamoeba dispar isolates in clinical samples by PCR and enzyme-linked immunosorbent assay. J Clin Microbiol 41:237–241. https://doi.org/10.1128/JCM.41.1.237-241.2003
Halliez MC, Buret AG (2015) Gastrointestinal parasites and the neural control of gut functions. Front Cell Neurosci 9:452. https://doi.org/10.3389/fncel.2015.00452
Henriksen SA, Pohlenz JF (1981) Staining of Cryptosporidia by modified Ziehl–Neelsen technique. Acta Vet Scand 22(3–4):594–596
Hunter PR, Nichols G (2002) Epidemiology and clinical features of Cryptosporidium infection in immunocompromised patients. Clin Microbiol Rev 15(1):145–154. https://doi.org/10.1128/CMR.15.1.145-154.2002
Insulander M, Silverlås C, Lebbad M, Karlsson L, Mattsson JG, Svenungsson B (2013) Molecular epidemiology and clinical manifestations of human cryptosporidiosis in Sweden. Epidemiol Infect 141(5):1009–1020. https://doi.org/10.1017/S0950268812001665
Jenkins MW, Maina-Gichaba C (2009) "Patterns and Sources of Faecal Pollution in the Heavily Impaired River Njoro Watershed Kenya: Findings and Implicaions". In: Proceedings of the Sumawa Mau forest complex conference. Nairobi—Kenya: Sumawa
Kenya Population and Housing Census (2019) Population by County and Sub County. Volume 1. http://housingfinanceafrica.org/app/uploads/VOLUME-I-KPHC-2019.pdf. Accessed 22 July 2021
Khalil S, Mirdha BR, Panda A, Singh Y, Makharia G, Paul J (2017) Cryptosporidium species subtypes and associated clinical manifestations in Indian patients. Gastroenterol Hepatol Bed to Bench 10(4):311–318
Khalil S, Mirdha BR, Paul J, Panda A, Singh Y (2018) Molecular detection and identification of Cryptosporidium viatorum in a human immunodeficiency virus-seropositive patient. J Glob Infect Dis 10:28–29. https://doi.org/10.4103/jgid_26_17
Kim K, Goozé L, Peterson C, Gut J, Nelson RG (1992) Isolation, sequence and molecular karyotype analysis of the actin gene of Cryptoporidium parvum. Mol Biochem Parasitol 50(1):105–113. https://doi.org/10.1016/0166-6851(92)90248-i
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
Koehler AV, Wang T, Haydon SR, Gasser RB (2018) Cryptosporidium viatorum from the native Australian swamp rat Rattus lutreolus: an emerging zoonotic pathogen? Int J Parasitol Parasites Wildl 7(1):18–26. https://doi.org/10.1016/j.ijppaw.2018.01.004
Laubach HE, Bentley CZ, Ginter EL, Spalter JS, Jensen LA (2004) A study of risk factors associated with the prevalence of Cryptosporidium in villages around Lake Atitlan. Guatemala Braz J Infect Dis 8(4):319–323. https://doi.org/10.1590/s1413-86702004000400008
Li N, Xiao L, Alderisio K, Elwin K, Cebelinski E, Chalmers R (2014) Subtyping Cryptosporidium ubiquitum, a zoonotic pathogen emerging in humans. Emerg Infect Dis 20(2):217–224. https://doi.org/10.3201/eid2002.121797
Liu A, Gong B, Liu X, Shen Y, Wu Y, Zhang W et al (2020) A retrospective epidemiological analysis of human Cryptosporidium infection in China during the past three decades, (1987–2018). PLoS Negl Trop Dis 14(3):e0008146. https://doi.org/10.1371/journal.pntd.0008146
Mainuri ZG, Owino JO (2013) Effects of land use and management on aggregate stability and hydraulic conductivity of soils within River Njoro watershed in Kenya. Int Soil Water Conserv Res 1(2):80–87. https://doi.org/10.1016/S2095-6339(15)30042-3
Mbae C, Mulinge E, Waruru A, Ngugi B, Wainaina J, Kariuki S (2015) Genetic diversity of Cryptosporidium in children in an urban informal settlement of Nairobi. Kenya Plos ONE 10(12):e0142055. https://doi.org/10.1371/journal.pone.0142055
Morse TD, Nichols RA, Grimason AM, Campbell BM, Tembo KC, Smith HV (2007) Incidence of cryptosporidiosis species in paediatric patients in Malawi. EpidemiolInfect 135(8):1307–1315. https://doi.org/10.1017/S0950268806007758
Pedraza-Diaz S, Amar C, Iversen AM, Stanley PJ, McLauchlin J (2001) Unusual Cryptosporidium species recovered from human feces: first description of Cryptosporidium felis and Cryptosporidium “dog type” from patients in England. J Med Microbiol 50(3):293–296. https://doi.org/10.1099/0022-1317-50-3-293
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
Putignani L, Menichella D (2010) Global distribution public health and clinical impact of the protozoan pathogen Cryptosporidium. Interdiscip Perspect Infect Dis. https://doi.org/10.1155/2010/753512
Quilez J, Sanchez-Acedo C, Avendaño C, del Cacho E, Lopez-Bernad F (2005) Efficacy of twoperoxygen-based disinfectants for inactivation of Cryptosporidium parvum oocysts. Appl Environ Microbiol 71(5):2479–2483. https://doi.org/10.1128/AEM.71.5.2479-2483.2005
Razakandrainibe R, Diawara EHI, Costa D, Le Goff L, Lemeteil D, Ballet JJ, Gargala G, Favennec L (2018) Common occurrence of Cryptosporidium hominis in asymptomatic and symptomatic calves in France. PLoS Negl Trop Dis 29(12(3)):e0006355
Robertson LJ, Johansen ØH, Kifleyohannes T, Efunshile AM, Terefe G (2020) Cryptosporidium infections in Africa-How Important is zoonotic transmission? a review of the evidence. Front Vet Sci 7:575881. https://doi.org/10.3389/fvets.2020.575881
Sánchez A, Munoz M, Gómez N, Tabares J, Segura L, Salazar Á et al (2017) Molecular epidemiology of Giardia, Blastocystis and Cryptosporidium among indigenous children from the colombian amazon basin. Front Microbiol 8:248. https://doi.org/10.3389/fmicb.2017.00248
Santos HL, Peralta RH, de Macedo HW, Barreto MG, Peralta JM (2007) Comparison of multiplex-PCR and antigen detection for differential diagnosis of Entamoeba histolytica. Braz J Infect Dis 11(3):365–370. https://doi.org/10.1590/s1413-86702007000300013
Shirley DA, Moonah SN, Kotloff KL (2012) Burden of disease from cryptosporidiosis. Curr Opin Infect Dis 25(5):555–563. https://doi.org/10.1097/QCO.0b013e328357e569
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 150:209–217. https://doi.org/10.1016/s0378-1097(97)00115-8
Squire SA, Yang R, Robertson I, Ayi I, Ryan U (2017) Molecular characterization of Cryptosporidium and Giardia in farmers and their ruminant livestock from the Coastal Savannah zone of Ghana. Infect Genet Evol 55:236–243. https://doi.org/10.1016/j.meegid.2017.09.025
Stensvold CR, Elwin K, Winiecka-Krusnell J, Chalmers RM, Xiao L, Lebbad M (2015) Development and application of a gp60-based typing assay for Cryptosporidium viatorum. J Clin Microbiol 53(6):1891–1897. https://doi.org/10.1128/JCM.00313-15
Sulaiman IM, Xiao L, Lal AA (1999) Evaluation of Cryptosporidium parvum genotyping techniques. Appl Environ Microbiol 65(10):4431–4435
Tumwine JK, Kekitiinwa A, Nabukeera N, Akiyoshi DE, Rich SM, Widmer G, Feng XC, Tzipori S (2003) Cryptosporidium parvum in children with diarrhea in Mulago Hospital, Kampala, Uganda. Am J Trop Med Hyg 68:710–715
Uga S, Matsuo J, Kono E, Kimura K, Inoue M, Rai SK, Ono K (2000) Prevalence of Cryptosporidium parvum infection and pattern of oocyst shedding in calves in Japan. Vet Parasitol 94(1–2):27–32. https://doi.org/10.1016/s0304-4017(00)00338-1
Ukwah BN, Ezeonu IM, Ezeonu CT, Roellig D, Xiao L (2017) Cryptosporidium species and subtypes in diarrheal children and HIV-infected persons in Ebonyi and Nsukka, Nigeria. J Infect Dev Ctries 11:173–179. https://doi.org/10.3855/jidc.8034
Walter EM, Charles M, Elick O, Manfred M, Domitila K (2021) Prevalence of zoonotic Cryptosporidium spp. isolates in Njoro Sub County, Nakuru County Kenya. Afr J Infect Dis 15(2):3–9. https://doi.org/10.21010/ajid.v15i2.2
Wang K, Gazizova A, Wang Y, Zhang K, Zhang Y, Chang Y (2019) First detection of Cryptosporidium spp. in migratory whooper swans (Cygnus cygnus) in China. Microorganisms 8(1):6. https://doi.org/10.3390/microorganisms8010006
Ward H (2019). Faculty opinions recommendation of life cycle progression and sexual development of the apicomplexan parasite Cryptosporidium parvum. Faculty Opinions – Post-
Wegayehu T, Karim MR, Li J, Adamu H, Erko B, Zhang L (2017) Prevalence and genetic characterization of Cryptosporidium species and Giardia duodenalis in lambs in Oromia special zone. Central Ethiopia BMC Vet Res 13(1):22. https://doi.org/10.1186/s12917-016-0916-0
Xiao L, Bern C, Limor J, Sulaiman I, Roberts J, Checkley W, Cabrera L, Gilman RH, Lal AA (2001) Identification of 5 types of Cryptosporidium parasites in children in Lima. Peru J Infect Dis 183(3):492–497. https://doi.org/10.1086/318090
Xu N, Liu H, Jiang Y, Yin J, Yuan Z, Shen Y (2020) First report of Cryptosporidium viatorum and Cryptosporidium occultus in humans in China, and of the unique novel C. viatorum subtype XVaA3h. BMC Infect Dis 20:16. https://doi.org/10.1186/s12879-019-4693-9
Yillia PT, Kreuzinger N, Mathooko JM (2008) The effect of in-stream activities on the Njoro River, Kenya. Part II: Microbial water quality. Phys Chem Earth 33(8–13):729–737. https://doi.org/10.1016/j.pce.2008.06.040
Zahedi A, Paparini A, Jian F, Robertson I, Ryan U (2016) Public health significance of zoonotic Cryptosporidium species in wildlife: critical insights into better drinking water management. Int J Parasitol Parasit Wildl 5(1):88–109. https://doi.org/10.1016/j.ijppaw.2015.12.001
Zhao W, Zhou H, Huang Y, Xu L, Rao L, Wang S et al (2019) Cryptosporidium spp. in wild rats (Rattus spp.) from the Hainan Province, China: Molecular detection, species/genotype identification and implications for public health. Int J Parasitol Parasit Wildl 9:317–321. https://doi.org/10.1016/j.ijppaw.2019.03.017
Acknowledgements
We thank Mr. Dickson Ayekha for his help during fieldwork and laboratory analysis. Special thanks go to the Biological Science Department, Egerton University, Kenya; Kenya Agricultural and Livestock Research Organization, Food Crops Research Centre Njoro, Kenya and the Inqaba Biotech Limited, Pretoria, South Africa for allowing us to use their facilities in sample analysis.
Funding
The authors did not receive financial support from any organization for the submitted work.
Author information
Authors and Affiliations
Contributions
Walter Miding’a Essendi, Charles Muleke and Elick Otachi contributed to the study concept and design. Material preparation, data collection and analysis were performed by Walter Miding’a Essendi and Manfred Miheso. The first draft of the manuscript was written by Walter Miding’a Essendi and all authors critically reviewed all versions of the manuscript. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that there is no conflict of interest and have no competing interest in this study.
Consent to participate
Informed consent was obtained from all individual participants included in the study.
Ethical approval
This study was conducted with approval provided by the Research and Ethical Committee of Egerton University (EUREC/APP/093/2019; Date: 4/3/2020).
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Essendi, W.M., Muleke, C., Miheso, M. et al. Genetic diversity of Cryptosporidium species in Njoro Sub County, Nakuru, Kenya. J Parasit Dis 46, 262–271 (2022). https://doi.org/10.1007/s12639-021-01444-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12639-021-01444-4