Abstract
Blastocystis sp. is a common zoonotic intestinal parasite of humans and animals, and has been classified into at least 17 distinct subtypes. Despite its potential impact on public health, the prevalence and subtype distribution of Blastocystis sp. have seldom been the study subject in South Korea. To determine the prevalence and subtype distribution of Blastocystis sp. and to obtain information on risk factors, we performed a cross-sectional study targeting elderly health checkup people, who visited Seoul Western Branch of the Korea Association of Health Promotion (KAHP) in October 2019. Stool samples were collected from 293 participants consisting of 128 males and 165 females with a mean age of 64.7 years (from 50 to 88 years) with a questionnaire on potential risk factors. All the samples were tested by PCR targeting the SSU rRNA gene of Blastocystis sp., and nucleotide sequences of positive samples were used to identify the subtypes. The overall prevalence of Blastocystis sp. was 9.2% (27/293). Among the positive samples, subtype 3 was predominant (59%; 16/27), followed by subtype 1 (41%; 11/27). No other subtypes were detected. In the univariable analysis, the age, sex, presence of digestive symptoms, source of drinking water, and history of drug intake were not significantly associated with Blastocystis sp. infection. Two parameters, including the Enterococcus hirae bacterial infection and the frequency of intake of cooked or boiled vegetables less than twice a week, showed statistical significance. However, the multivariable regression analysis revealed that only the latter parameter was statistically significant. The results suggested that subtypes 3 and 1 are the 2 major genotypes of Blastocystis sp. among elderly people in South Korea, and low frequency of consuming cooked or boiled vegetables is a potential risk factor.
Similar content being viewed by others
References
Alfellani MA, Stensvold CR, Vidal-Lapiedra A, Onuoha ES, Fagbenro-Beyioku AF, Clark CG (2013a) Variable geographic distribution of Blastocystis subtypes and its potential implications. Acta Trop 126:11–18. https://doi.org/10.1016/j.actatropica.2012.12.011
Alfellani MA, Taner-Mulla D, Jacob AS, Imeede CA, Yoshikawa H, Stendvold CR, Clark CG (2013b) Genetic diversity of Blastocystis in livestock and zoo animals. Protist 164:497–509. https://doi.org/10.1016/j.protis.2013.05.003
Andersen LO, Bonde I, Nielsen HB, Stensvold CR (2015) A retrospective metagenomics approach to studying Blastocystis. FEMS Microbiol Ecol 91:1–9. https://doi.org/10.1093/femsec/fiv072
Beghini F, Pasolli E, Truong TD, Putignani L, Caccio SM, Segata N (2017) Large-scale comparative metagenomics of Blastocystis, a common member of the human gut microbiome. ISME J 11:2848–2863. https://doi.org/10.1038/ismej.2017.139
Boorom KF, Smith H, Nimri L, Viscogliosi E, Spanakos G, Parkar U, Li LH, Zhou XN, Ok UZ, Leelayoova S, Jones MS (2008) Oh my aching gut: irritable bowel syndrome, Blastocystis, and asymptomatic infection. Parasit Vectors 1:40. https://doi.org/10.1186/1756-3305-1-40
Chan TS, Wu MS, Suk FM, Chen CN, Chen YF, Hou YH, Lien GS (2012) Enterococcus hirae-related acute pyelonephritis and cholangitis with bacteremia: an unusual infection in humans. Kaohsiung J Med Sci 28:111–114. https://doi.org/10.1016/j.kjms.2011.06.027
Chen TL, Chan CC, Chen HP, Fung CP, Lin CP, Chan WL, Liu CY (2003) Clinical characteristics and endoscopic findings associated with Blastocystis hominis in healthy adults. Am J Trop Med Hyg 69:213–216. https://doi.org/10.4269/ajtmh.2003.69.213
Chenoweth C, Schaberg D (1990) The epidemiology of enterococci. Eur J Clin Microbiol Infect Dis 9:80–89. https://doi.org/10.1007/BF01963631
Clark CG, van der Giezen M, Alfellani MA, Stensvold CR (2013) Recent developments in Blastocystis research. Adv Parasitol 82:1–32. https://doi.org/10.1016/B978-0-12-407706-5.00001-0
Deng L, Chai Y, Zhou Z, Liu H, Zhong Z, Hu Y, Fu H, Yue C, Peng G (2019) Epidemiology of Blastocystis sp. infection in China: a systematic review. Parasite 26:41. https://doi.org/10.1051/parasite/2019042
El Safadi D, Cian A, Nourrisson C, Pereira B, Morelle C, Bastien P, Bellanger AP, Botterel F, Candolfi E, Desoubeaux G, Lachaud L, Morio F, Pomares C, Rabodonirina M, Wawrzyniak I, Delbac F, Gantois N, Certad G, Delhaes L, Poirier P, Viscogliosi E. (2016) Prevalence, risk factors for infection and subtype distribution of the intestinal parasite Blastocystis sp. from a large-scale multi-center study in France. BMC Infect Dis 16:451. https://doi.org/10.1186/s12879-016-1776-8
Haider SS, Baqai R, Qureshi FM, Boorom K (2012) Blastocystis spp., Cryptosporidium spp., and Entamoeba histolytica exhibit similar symptomatic and epidemiological patterns in healthcare-seeking patients in Karachi. Parasitol Res 111:1357–1368. https://doi.org/10.1007/s00436-012-2972-0
Hirata T, Nakamura H, Kinjo N, Hokama A, Kinjo F, Yamane N, Fujita J (2007) Prevalence of Blastocystis hominis and Strongyloides stercoralis infection in Okinawa, Japan. Parasitol Res 101:1717–1719. https://doi.org/10.1007/s00436-007-0712-7
Iebba V, Santangelo F, Totino V, Pantanella F, Monsia A, Cristanziano VD, Cave DD, Schippa S, Berrilli F, D’Alfonso R (2016) Gut microbiota related to Giardia duodenalis, Entamoeba spp. and Blastocystis hominis infections in humans from Cote d’Ivoire. J Infect Dev Ctries 10:1035–1041. https://doi.org/10.3855/jidc.8179
Javanmard E, Rahimi HM, Niyyati M, Aghdaei HA, Sharifdini M, Mirjalali H, Zali MR, Karanis P (2019) Molecular analysis of Blastocystis sp. and its subtypes from treated wastewater routinely used for irrigation of vegetable farmlands in Iran. J Water Health 17:837–844. https://doi.org/10.2166/wh.2019.045
McNamee R (2003) Confounding and confounders. Occup Environ Med 60:227–234. https://doi.org/10.1136/oem.60.3.227
Kim MJ, Won EJ, Kim SH, Shin JH, Chai JY (2020a) Molecular detection and subtyping of human Blastocystis and the clinical implications: comparisons between diarrheal and non-diarrheal groups in Korean populations. Korean J Parasitol 58:321–326. https://doi.org/10.3347/kjp.2020.58.3.321
Kim NH, Kim HW, Park SM, Seo GH, Cho TJ, Yu HR, Kim SH, Hwang JH, Choi C, Rhee MS (2020b) Virulence patterns and prevalence of seven Enterococcus species isolated from meats and leafy vegetables in South Korea. Food Control 108:106867. https://doi.org/10.1016/j.foodcont.2019.106867
Kim SH, Kwon DY, Shin D (2020c) Namul, the deiving force behind health and high vegetable consumption in Korea. J Ethnic Foods 7:15. https://doi.org/10.1186/s42779-019-0026-2
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948. https://doi.org/10.1093/bioinformatics/btm404
Lee LI, Chye TT, Karmacharya BM, Govind SK (2012) Blastocystis sp.: waterborne zoonotic organism, a possibility? Parasit Vectors 5:130. https://doi.org/10.1186/1756-3305-5-130
Leelayoova S, Siripattanapipong S, Thathaisong U, Naaglor T, Taamasri P, Piyaraj P, Mungthin M (2008) Drinking water: a possible source of Blastocystis spp. subtype 1 infection in schoolchildren of a rural community in central Thailand. Am J Trop Med Hyg 79:401–406. https://doi.org/10.4269/ajtmh.2008.79.401
Mattiucci S, Crisafi B, Gabrielli S, Paoletti M, Cancrini G (2016) Molecular epidemiology and genetic diversity of Blastocystis infection in humans in Italy. Epidemiol Infect 144:635–646. https://doi.org/10.1017/S0950268815001697
Nieves-Ramirez ME, Partida-Rodríguez O, Laforest-Lapointe I, Reynolds LA, Brown EM, Valdez-Salazar A, Morán-Silva P, Rojas-Velázquez L, Morien E, Parfrey LW, Jin M, Walter J, Torres J, Arrieta MC, Ximénez-García C, Finlay BB (2018) Asymptomatic intestinal colonization with protist Blastocystis is strongly associated with distinct microbiome ecological patterns. mSystems 3:e00007–18. https://doi.org/10.1128/mSystems.00007-18
Parkar U, Traub RJ, Kumar S, Mungthin M, Vitali S, Leelayoova S, Morris K, Thompson RCA (2007) Direct characterization of Blastocystis from faeces by PCR and evidence of zoonotic potential. Parasitology 134(Pt 3):359–367. https://doi.org/10.1017/S0031182006001582
Pipatsatitpong D, Rangsin R, Leelayoova S, Naaglor T, Mungthin M (2012) Incidence and risk factors of Blastocystis infection in an orphanage in Bangkok. Thailand Parasit Vectors 5:37. https://doi.org/10.1186/1756-3305-5-37
Poirier P, Wawrzyniak I, Albert A, El Alaoui H, Delbac F, Livrelli V (2011) Development and evaluation of a real-time PCR assay for detection and quantification of Blastocystis parasites in human stool samples: prospective study of patients with hematological malignancies. J Clin Microbiol 49:975–983. https://doi.org/10.1128/JCM.01392-10
Pourhoseingholi MA, Baghestani AR, Vahedi M (2012) How to control confounding effects by statistical analysis. Gastroenterol Hepatol Bed Bench 5(2):79–83
Ramirez JD, Sanchez LV, Bautista DC, Corredor AF, Florez AC, Stensvold CR (2014) Blastocystis subtypes detected in humans and animals from Colombia. Infect Genet Evol 22:223–228. https://doi.org/10.1016/j.meegid.2013.07.020
Roberts T, Barratt J, Harkness J, Ellis J, Stark D (2011) Comparison of microscopy, culture, and conventional polymerase chain reaction for detection of Blastocystis sp. in clinical stool samples. Am J Trop Med Hyg 84:308–312. https://doi.org/10.4269/ajtmh.2011.10-0447
Salem-Bekhit MM, Moussa IM, Muharram MM, Alanazy FK, Hefni HM (2012) Prevalence and antimicrobial resistance pattern of multidrug-resistant enterococci isolated from clinical specimens. Indian J Med Microbiol 30:44–51. https://doi.org/10.4103/0255-0857.93032
Santin M, Gomez-Munoz MT, Solano-Aguilar G, Fayer R (2011) Development of a new PCR protocol to detect and subtype Blastocystis spp. from humans and animals. Parasitol Res 109:205–212. https://doi.org/10.1007/s00436-010-2244-9
Scicluna SM, Tawari B, Clark CG (2006) DNA barcoding of Blastocystis. Protist 157:77–85. https://doi.org/10.1016/j.protis.2005.12.001
Stark D, van Hal S, Marriott D, Ellis J, Harkness J (2007) Irritable bowel syndrome: a review on the role of intestinal protozoa and the importance of their detection and diagnosis. Int J Parasitol 37:11–20. https://doi.org/10.1016/j.ijpara.2006.09.009
Stensvold CR, Clark CG (2020) Pre-empting Pandora’s box: Blastocystis subtypes revisited. Trends Parasitol 36:229–232. https://doi.org/10.1016/j.pt.2019.12.009
Stensvold CR, Suresh GK, Kevin SW, Tan RC, Thompson A, Traub RJ, Viscogliosi E, Yoshikawa H, Clark CG (2007) Terminology for Blastocystis subtypes-a consensus. Trends Parasitol 23:93–96. https://doi.org/10.1016/j.pt.2007.01.004
Stensvold CR, Alfellani MA, Nørskov-Lauritsen S, Prip K, Victory EL, Maddox C, Nielsen HV, Clark CG (2009) Subtype distribution of Blastocystis isolates from synanthropic and zoo animals and identification of a new subtype. Int J Parasitol 39:473–479. https://doi.org/10.1016/j.ijpara.2008.07.006
Stensvold CR, Tan KSW, Clark CG (2020) Blastocystis. Trends Parasitol 36:315–316. https://doi.org/10.1016/j.pt.2019.12.008
Su FH, Chu FY, Li CY, Tang HF, Lin YS, Peng YJ, Su YM, Lee SD (2009) Blastocystis hominis infection in long-term care facilities in Taiwan: prevalence and associated clinical factors. Parasitol Res 105:1007–1013. https://doi.org/10.1007/s00436-009-1509-7
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S (2013) MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Mol Biol Evol 30:2725–2729. https://doi.org/10.1093/molbev/mst197
Tan KS (2008) New insights on classification, identification, and clinical relevance of Blastocystis spp. Clin Microbiol Rev 21(4):639–665. https://doi.org/10.1128/CMR.00022-08
Tasova Y, Sahin B, Koltas S, Paydas S (2000) Clinical significance and frequency of Blastocystis hominis in Turkish patients with hematological malignancy. Acta Med Okayama 54:133–136. https://doi.org/10.18926/AMO/32298
Verweij JJ, Laeijendecker D, Brienen EA, van Lieshout L, Polderman AM (2003) Detection of Cyclospora cayetanensis in travellers returning from the tropics and subtropics using microscopy and real-time PCR. Int J Med Microbiol 293:199–202. https://doi.org/10.1078/1438-4221-00252
Wawrzyniak I, Poirier P, Viscogliosi E, Dionigia M, Texier C, Delbac F, Alaoui HE (2013) Blastocystis, an unrecognized parasite: an overview of pathogenesis and diagnosis. Ther Adv Infect Dis 1:167–178. https://doi.org/10.1177/2049936113504754
Won EJ, Kim SH, Kee SJ, Shin JH, Suh SP, Chai JY, Ryang DW, Shin MG (2016) Multiplex real-time PCR assay targeting eight parasites customized to the Korean population: potential use for detection in diarrheal stool samples from gastroenteritis patients. PLoS ONE 11:e0166957. https://doi.org/10.1371/journal.pone.0166957
Xiao L, Morgan UM, Limor J, Escalante A, Arrowood M, Shulaw W, Thompson RC, Fayer R, Lal AA (1999) Genetic diversity within Cryptosporidium parvum and related Cryptosporidium species. Appl Environ Microbiol 65:3386–3391. https://doi.org/10.1128/AEM.65.8.3386-3391.1999
Yason JA, Liang YR, Png CW, Zhang Y, Tan KSW (2019) Interactions between a pathogenic Blastocystis subtype and gut microbiota: in vitro and in vivo studies. Microbiome 7:30. https://doi.org/10.1186/s40168-019-0644-3
Acknowledgements
The authors thank the staff of the Seoul Western Branch, Korea Association of Health Promotion, Seoul, South Korea, who helped with sample collection. We also appreciate the staff in the Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, South Korea, who helped with molecular analysis of Enterococcus spp.
Funding
This study was supported by a grant from the Ministry of Food and Drug Safety, the Republic of Korea, in 2017–2019 (17162MFDS034).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethical approval
The procedures of the present study were conducted following the guidelines and approval of the Institutional Review Board of Korea Association of Health Promotion (IRB no. 130750–201902-BR-019).
Conflict of interest
The authors declare no competing interests.
Additional information
Section Editor: Yaoyu Feng
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
About this article
Cite this article
Chang, T., Jung, BK., Shin, H. et al. Genotypes of Blastocystis sp. among elderly health checkup people in South Korea with a questionnaire on risk factors. Parasitol Res 120, 3297–3306 (2021). https://doi.org/10.1007/s00436-021-07274-w
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00436-021-07274-w