Epidemiological and Molecular Characterization of Blastocystis Infection in Children Attending Daycare Centers in Medellín, Colombia
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Study Population
2.2. Sociodemographic Variables
2.3. Parasitological Diagnosis
2.4. Molecular Detection of Blastocystis
2.5. Subtype Identification and Phylogenetic Analyses
2.6. Statistical Analysis
3. Results
3.1. Study Population
3.2. Infection with Intestinal Parasites
3.3. Coinfection by Microscopy
3.4. Molecular Characterization of Blastocystis
3.5. Association between Sociodemographic Variables and Intestinal Parasites
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Tan, K.S.W. New Insights on Classification, Identification, and Clinical Relevance of Blastocystis spp. Clin. Microbiol. Rev. 2008, 21, 639–665. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Boorom, K.F.; Smith, H.; Nimri, L.; Viscogliosi, E.; Spanakos, G.; Parkar, U.; Li, L.-H.; Zhou, X.-N.; Ok, Ü.Z.; Leelayoova, S.; et al. Oh my aching gut: Irritable bowel syndrome, Blastocystis, and asymptomatic infection. Parasit. Vectors 2008, 1, 40. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Scanlan, P.D.; Stensvold, C.R. Blastocystis: Getting to grips with our guileful guest. Trends Parasitol. 2013, 29, 523–529. [Google Scholar] [CrossRef]
- MinSalud. MinSalud Encuesta Nacional de Parasitismo Intestinal en Población Escolar Colombia. 2015. Available online: https://www.minsalud.gov.co/sites/rid/Lists/BibliotecaDigital/RIDE/VS/PP/ET/encuesta-nacional-de-parasitismo-2012-2014.pdf (accessed on 1 February 2021).
- Lucero-Garzón, T.A.; Álvarez-Motta, L.A.; Chicue-López, J.F. Parasitosis intestinal y factores de riesgo en niños de los asentamientos subnormales, Florencia-Caquetá, Colombia. Rev. Fac. Nac. Salud. Pública. 2015, 33, 171–180. [Google Scholar] [CrossRef] [Green Version]
- Londoño, Á.; Mejía, S.; Gómez-Marín, J. Prevalencia y Factores de Riesgo Asociados a Parasitismo Intestinal en Preescolares de Zona Urbana en Calarcá, Colombia. Rev. Salud pública. 2009, 11, 72–81. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Pedraza, B.; Suarez, H.; De-la-Hoz, I.; Fragoso, P. Parasitosis intestinal relacionada con el estado nutricional de los niños de 2 a 5 años en hogares comunitarios de Cartagena de Indias, Colombia. Rev. Chil. Nutr. 2019, 46, 239–244. [Google Scholar] [CrossRef] [Green Version]
- Higuera, A.; Villamizar, X.; Herrera, G.; Giraldo, J.C.; Vasquez-A, L.R.; Urbano, P.; Villalobos, O.; Tovar, C.; Ramírez, J.D. Molecular detection and genotyping of intestinal protozoa from different biogeographical regions of Colombia. PeerJ 2020, 8, e8554. [Google Scholar] [CrossRef] [Green Version]
- Cenac, N.; Andrews, C.N.; Holzhausen, M.; Chapman, K.; Cottrell, G.; Andrade-Gordon, P.; Steinhoff, M.; Barbara, G.; Beck, P.; Bunnett, N.W.; et al. Role for protease activity in visceral pain in irritable bowel syndrome. J. Clin. Invest. 2007, 117, 636–647. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Fouad, S.A.; Basyoni, M.M.A.; Fahmy, R.A.; Kobaisi, M.H. The pathogenic role of different Blastocystis hominis genotypes isolated from patients with irritable bowel syndrome. Arab J. Gastroenterol. 2011, 12, 194–200. [Google Scholar] [CrossRef]
- Nourrisson, C.; Scanzi, J.; Pereira, B.; NkoudMongo, C.; Wawrzyniak, I.; Cian, A.; Viscogliosi, E.; Livrelli, V.; Delbac, F.; Dapoigny, M.; et al. Blastocystis Is Associated with Decrease of Fecal Microbiota Protective Bacteria: Comparative Analysis between Patients with Irritable Bowel Syndrome and Control Subjects. PLoS ONE 2014, 9, e111868. [Google Scholar] [CrossRef]
- Nasirudeen, A.M.A.; Tan, K.S.W. Isolation and characterization of the mitochondrion-like organelle from Blastocystis hominis. J. Microbiol. Methods 2004, 58, 101–109. [Google Scholar] [CrossRef] [PubMed]
- Denoeud, F.; Roussel, M.; Noel, B.; Wawrzyniak, I.; Da Silva, C.; Diogon, M.; Viscogliosi, E.; Brochier-Armanet, C.; Couloux, A.; Poulain, J.; et al. Genome sequence of the stramenopile Blastocystis, a human anaerobic parasite. Genome Biol. 2011, 12, R29. [Google Scholar] [CrossRef] [PubMed]
- del Coco, V.F.; Molina, N.B.; Basualdo, J.A.; Córdoba, M.A. Blastocystis spp.: Avances, controversias y desafíos futuros. Rev. Argent. Microbiol. 2017, 49, 110–118. [Google Scholar] [CrossRef] [PubMed]
- Jiménez, P.A.; Jaimes, J.E.; Ramírez, J.D. A summary of Blastocystis subtypes in North and South America. Parasit. Vectors 2019, 12, 376. [Google Scholar] [CrossRef]
- Ramírez, J.D.; Sánchez, A.; Hernández, C.; Flórez, C.; Bernal, M.C.; Giraldo, J.C.; Reyes, P.; López, M.C.; García, L.; Cooper, P.J.; et al. Geographic distribution of human Blastocystis subtypes in South America. Infect. Genet. Evol. 2016, 41, 32–35. [Google Scholar] [CrossRef]
- Ramírez, J.D.; Flórez, C.; Olivera, M.; Bernal, M.C.; Giraldo, J.C. Blastocystis subtyping and its association with intestinal parasites in children from different geographical regions of Colombia. PLoS ONE 2017, 12, e0172586. [Google Scholar] [CrossRef] [Green Version]
- Ramírez, J.D.; Sánchez, L.V.; Bautista, D.C.; Corredor, A.F.; Flórez, A.C.; Stensvold, C.R. Blastocystis subtypes detected in humans and animals from Colombia. Infect. Genet. Evol. 2014, 22, 223–228. [Google Scholar] [CrossRef]
- Sankur, F.; Ayturan, S.; Malatyali, E.; Ertabaklar, H.; Ertug, S. The Distribution of Blastocystis Subtypes among School-aged Children in Mugla, Turkey. Iran. J. Parasitol. 2017, 12, 580–586. [Google Scholar]
- Dogan, N.; Aydin, M.; Tuzemen, N.U.; Dinleyici, E.C.; Oguz, I.; Dogruman-Al, F. Subtype distribution of Blastocystis spp. isolated from children in Eskisehir, Turkey. Parasitol. Int. 2017, 66, 948–951. [Google Scholar] [CrossRef]
- Stensvold, C.R.; Arendrup, M.C.; Jespersgaard, C.; Mølbak, K.; Nielsen, H.V. Detecting Blastocystis using parasitologic and DNA-based methods: A comparative study. Diagn. Microbiol. Infect. Dis. 2007, 59, 303–307. [Google Scholar] [CrossRef] [PubMed]
- Khademvatan, S.; Masjedizadeh, R.; Yousefi-Razin, E.; Mahbodfar, H.; Rahim, F.; Yousefi, E.; Foroutan, M. PCR-based molecular characterization of Blastocystis hominis subtypes in southwest of Iran. J. Infect. Public Health 2018, 11, 43–47. [Google Scholar] [CrossRef]
- Menounos, P.G.; Spanakos, G.; Tegos, N.; Vassalos, C.M.; Papadopoulou, C.; Vakalis, N.C. Direct detection of Blastocystis sp. in human faecal samples and subtype assignment using single strand conformational polymorphism and sequencing. Mol. Cell. Probes 2008, 22, 24–29. [Google Scholar] [CrossRef]
- Villamizar, X.; Higuera, A.; Herrera, G.; Vasquez-A, L.R.; Buitron, L.; Muñoz, L.M.; Gonzalez-C, F.E.; Lopez, M.C.; Giraldo, J.C.; Ramírez, J.D. Molecular and descriptive epidemiology of intestinal protozoan parasites of children and their pets in Cauca, Colombia: A cross-sectional study. BMC Infect. Dis. 2019, 19, 190. [Google Scholar] [CrossRef] [Green Version]
- Sánchez, A.; Munoz, M.; Gómez, N.; Tabares, J.; Segura, L.; Salazar, Á.; Restrepo, C.; Ruíz, M.; Reyes, P.; Qian, Y.; et al. Molecular Epidemiology of Giardia, Blastocystis and Cryptosporidium among Indigenous Children from the Colombian Amazon Basin. Front. Microbiol. 2017, 8. [Google Scholar] [CrossRef] [PubMed]
- Restrepo, C.; Berrio, L.; Salazar, M. Evaluación de tres técnicas coproparasitoscópicas para el diagnóstico de geohelmintos intestinales. Latreia 2012, 26, 15–24. [Google Scholar]
- Stensvold, C.R.; Ahmed, U.N.; Andersen, L.O.; Nielsen, H.V. Development and evaluation of a genus-specific, probe-based, internal-process-controlled real-time PCR assay for sensitive and specific detection of Blastocystis spp. J. Clin. Microbiol. 2012, 50, 1847–1851. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Scicluna, S.M.; Tawari, B.; Clark, C.G. DNA barcoding of blastocystis. Protist 2006, 157, 77–85. [Google Scholar] [CrossRef]
- Katoh, K.; Standley, D.M. MAFFT Multiple Sequence Alignment Software Version 7: Improvements in Performance and Usability. Mol. Biol. Evol. 2013, 30, 772–780. [Google Scholar] [CrossRef] [Green Version]
- Price, M.N.; Dehal, P.S.; Arkin, A.P. FastTree: Computing Large Minimum Evolution Trees with Profiles instead of a Distance Matrix. Mol. Biol. Evol. 2009, 26, 1641–1650. [Google Scholar] [CrossRef] [PubMed]
- Letunic, I.; Bork, P. Interactive Tree Of Life (iTOL) v4: Recent updates and new developments. Nucleic Acids Res. 2019, 47, W256–W259. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Wawrzyniak, I.; Poirier, P.; Viscogliosi, E.; Dionigia, M.; Texier, C.; Delbac, F.; Alaoui, H.E. Blastocystis, an unrecognized parasite: An overview of pathogenesis and diagnosis. Ther. Adv. Infect. Dis. 2013, 1, 167–178. [Google Scholar] [CrossRef] [Green Version]
- Alinaghizade, A.; Mirjalali, H.; Mohebali, M.; Stensvold, C.R.; Rezaeian, M. Inter- and intra-subtype variation of Blastocystis subtypes isolated from diarrheic and non-diarrheic patients in Iran. Infect. Genet. Evol. 2017, 50, 77–82. [Google Scholar] [CrossRef]
- Dacal, E.; Saugar, J.M.; de Lucio, A.; Hernández-de-Mingo, M.; Robinson, E.; Köster, P.C.; Aznar-Ruiz-de-Alegría, M.L.; Espasa, M.; Ninda, A.; Gandasegui, J.; et al. Prevalence and molecular characterization of Strongyloides stercoralis, Giardia duodenalis, Cryptosporidium spp., and Blastocystis spp. isolates in school children in Cubal, Western Angola. Parasit. Vectors 2018, 11, 1. [Google Scholar] [CrossRef] [Green Version]
- Leelayoova, S.; Mungthin, M.; Aunpad, R.; Naaglor, T.; Rangsin, R.; Pipatsatitpong, D. Prevalence and Risk Factors for Blastocystis Infection Among Children and Caregivers in a Child Care Center, Bangkok, Thailand. Am. J. Trop. Med. Hyg. 2015, 93, 310–315. [Google Scholar]
- Scanlan, P.D.; Knight, R.; Song, S.J.; Ackermann, G.; Cotter, P.D. Prevalence and genetic diversity of Blastocystis in family units living in the United States. Infect. Genet. Evol. 2016, 45, 95–97. [Google Scholar] [CrossRef] [PubMed]
- Oliveira-Arbex, A.P.; David, É.B.; Guimarães, S. Blastocystis genetic diversity among children of low-income daycare center in Southeastern Brazil. Infect. Genet. Evol. 2018, 57, 59–63. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Perea, M.; Vásquez, V.; Pineda, V.; Samudio, F.; Calzada, J.E.; Saldaña, A. Prevalence and subtype distribution of Blastocystis sp. infecting children from a rural community in Panama. Parasite Epidemiol. Control. 2020, 9, e00139. [Google Scholar] [CrossRef] [PubMed]
- Hernández, P.C.; Morales, L.; Chaparro-Olaya, J.; Sarmiento, D.; Jaramillo, J.F.; Ordoñez, G.A.; Cortés, F.; Sánchez, L.K. Intestinal parasitic infections and associated factors in children of three rural schools in Colombia. A cross-sectional study. PLoS ONE 2019, 14, e0218681. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Medina-A, B.J. Prevalencia de parásitos intestinales en niños que asisten al templo comedor Sagrado Corazón Teresa Benedicta de La Cruz del Barrio Vallejuelos, Medellín 2007. Rev. Iatreia. 2009, 22, 227–234. [Google Scholar]
- Bryan, P.E.; Romero, M.; Sánchez, M.; Torres, G.; Gómez, W.; Restrepo, M.; Restrepo, A.; Mejia, R. Urban versus Rural Prevalence of Intestinal Parasites Using Multi-Parallel qPCR in Colombia. Am. J. Trop. Med. Hyg. 2020. [Google Scholar] [CrossRef]
- Naceanceno, K.S.; Matamoros, G.; Gabrie, J.A.; Bottazzi, M.E.; Sanchez, A.; Mejia, R. Use of Multi-Parallel Real-Time Quantitative PCR to Determine Blastocystis Prevalence and Association with Other Gastrointestinal Parasite Infection in a Rural Honduran Location. Am. J. Trop. Med. Hyg. 2020, 102, 1373–1375. [Google Scholar] [CrossRef]
- Vaca, M.; Cooper, P.J.; Cortés, A.A.; Chico, M.; Sandoval, C.; Weatherhead, J.; Mejia, R.; Loor, S. Comparison of Cytokine Responses in Ecuadorian Children Infected with Giardia, Ascaris, or Both Parasites. Am. J. Trop. Med. Hyg. 2017, 96, 1394–1399. [Google Scholar]
- Supali, T.; Verweij, J.J.; Wiria, A.E.; Djuardi, Y.; Hamid, F.; Kaisar, M.M.M.; Wammes, L.J.; van Lieshout, L.; Luty, A.J.F.; Sartono, E.; et al. Polyparasitism and its impact on the immune system. Int. J. Parasitol. 2010, 40, 1171–1176. [Google Scholar] [CrossRef]
- Morgan, U.M.; Pallant, L.; Dwyer, B.W.; Forbes, D.A.; Rich, G.; Thompson, R.C.A. Comparison of PCR and Microscopy for Detection of Cryptosporidium parvum in Human Fecal Specimens: Clinical Trial. J. Clin. Microbiol. 1998, 36, 995–998. [Google Scholar] [CrossRef] [Green Version]
- Stensvold, C.R.; Winiecka-Krusnell, J.; Lier, T.; Lebbad, M. Evaluation of a PCR Method for Detection of Entamoeba polecki, with an Overview of Its Molecular Epidemiology. J. Clin. Microbiol. 2018, 56, e00154-18. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Carvajal-Restrepo, H.; Orrego-Morales, C.; Vega-Orrego, T.; Arango-Arango, S.; Buitrago-Agudelo, D.; Maya-Betancourt, M.C.; Maya-Betancourt, V.; Restrepo-Álvarez, L.; Silva-Cáceres, N.; Suarez-Urquijo, S.; et al. Screening for intestinal parasites in adults from three different regions of Colombia. Infectio 2018, 23, 33–38. [Google Scholar] [CrossRef]
- Agudelo-Lopez, S.; Gómez-Rodríguez, L.; Coronado, X.; Orozco, A.; Valencia-Gutierrez, C.A.; Restrepo-Betancur, L.F.; Galvis-Gómez, L.A.; Botero-Palacio, L.E. Prevalencia de Parasitosis Intestinales y Factores Asociados en un Corregimiento de la Costa Atlántica Colombiana. Rev. Salud Pública 2008, 10, 633–642. [Google Scholar] [CrossRef] [Green Version]
- Maloney, J.G.; Molokin, A.; da Cunha, M.J.R.; Cury, M.C.; Santin, M. Blastocystis subtype distribution in domestic and captive wild bird species from Brazil using next generation amplicon sequencing. Parasite Epidemiol. Control 2020, 9, e00138. [Google Scholar] [CrossRef]
- Greige, S.; El Safadi, D.; Bécu, N.; Gantois, N.; Pereira, B.; Chabé, M.; Benamrouz-Vanneste, S.; Certad, G.; El Hage, R.; Chemaly, M.; et al. Prevalence and subtype distribution of Blastocystis sp. isolates from poultry in Lebanon and evidence of zoonotic potential. Parasit. Vectors 2018, 11, 389. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Alfellani, M.A.; Taner-Mulla, D.; Jacob, A.S.; Imeede, C.A.; Yoshikawa, H.; Stensvold, C.R.; Clark, C.G. Genetic Diversity of Blastocystis in Livestock and Zoo Animals. Protist 2013, 164, 497–509. [Google Scholar] [CrossRef] [Green Version]
- Betts, E.L.; Gentekaki, E.; Tsaousis, A.D. Exploring Micro-Eukaryotic Diversity in the Gut: Co-occurrence of Blastocystis Subtypes and Other Protists in Zoo Animals. Front. Microbiol. 2020, 11, 288. [Google Scholar] [CrossRef] [PubMed]
- Zanetti, A.d.S.; Malheiros, A.F.; de Matos, T.A.; Longhi, F.G.; Moreira, L.M.; Silva, S.L.; Castrillon, S.K.I.; Ferreira, S.M.B.; Ignotti, E.; Espinosa, O.A. Prevalence of Blastocystis sp. infection in several hosts in Brazil: A systematic review and meta-analysis. Parasit. Vectors 2020, 13, 30. [Google Scholar] [CrossRef] [PubMed]
- Valença-Barbosa, C.; do Bomfim, T.C.B.; Teixeira, B.R.; Gentile, R.; Neto, S.F.d.C.; Magalhães, B.S.N.; Balthazar, D.d.A.; da Silva, F.A.; Biot, R.; d’Avila Levy, C.M.; et al. Molecular epidemiology of Blastocystis isolated from animals in the state of Rio de Janeiro, Brazil. PLoS ONE 2019, 14, e0210740. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Olsen, K.E.P.; Christiansen, D.B.; Nielsen, H.V.; Stensvold, C.R. Blastocystis sp. Subtype 4 is Common in Danish Blastocystis-Positive Patients Presenting with Acute Diarrhea. Am. J. Trop. Med. Hyg. 2011, 84, 883–885. [Google Scholar] [CrossRef]
- Gaviria, L.M.; Soscue, D.; Campo-Polanco, L.F.; Cardona-Arias, J.; Galván-Díaz, A.L. Prevalencia de parasitosis intestinal, anemia y desnutrición en niños de un resguardo indígena Nasa, Cauca, Colombia, 2015. Rev. Fac. Nac. Salud Pública 2017, 35, 390–399. [Google Scholar] [CrossRef] [Green Version]
Intestinal Protozoa | Diagnostic Method | ||
---|---|---|---|
Direct | Concentration | Prevalence | |
Protozoa | |||
Blastocystis | 14.7% (39/265) | 11.7% (31/265) | 15.8% (42/265) |
Giardia intestinalis | 12.5% (33/265) | 15.1% (40/265) | 15.5% (41/265) |
Endolimax nana | 10.6% (28/265) | 9.15% (24/265) | 15.1% (40/265) |
Entamoeba coli | 4.2% (11/265) | 5.7% (15/265) | 6% (16/265) |
Entamoeba complex | 1.1% (3/265) | 0.8% (2/265) | 1.1% (3/265) |
Iodamoeba butschlii | 0.8% (2/265) | 0.8% (2/265) | |
Chilomastix mesnili | 0.8% (2/265) | 1.1% (3/265) | 1.1% (3/265) |
Helminths | |||
Ascaris lumbricoides | 0.8% (2/265) | 0.8% (2/265) | 0.8% (2/265) |
Variable | Category | Intestinal Parasites | ||||
---|---|---|---|---|---|---|
Positive | Negative | Total | OR (CI 95%) | p-Value | ||
Sex | Male | 42 (31.6%) | 91 | 133 | 0.6 (0.391–1.069) | 0.088 |
Female | 55 (41.6%) | 77 | 132 | |||
Age | >30 months | 88 (58.6%) | 62 | 150 | 0.5 (0.346–0.970) | 0.042 |
<30 months | 53 (46.1%) | 62 | 115 | |||
Nationality | Venezuelan | 13 (52%) | 12 | 25 | 2.0 (0.926–4.732) | 0.108 |
Colombian | 79 (35.6%) | 143 | 222 | |||
Type of population | Rural | 33 (51.6%) | 31 | 64 | 2.2 (1.285–4.042) | 0.004 |
Urban | 64 (31.8%) | 137 | 201 | |||
Children’s center | 2 * | 33 (52.3%) | 30 | 63 | 3.9 (2.267–6.711) | 0.000 |
1, 3, 4, 5 ** | 64 (31.6%) | 138 | 202 | |||
Health affiliation scheme | Subsidized | 61 (48%) | 66 | 127 | 2.6 (1.551–4.539) | 0.000 |
Contributory | 31 (25.8%) | 89 | 120 | |||
Stratum | 1-2 | 73 (38.2%) | 118 | 191 | 1.2 (0.645–2.252) | 0.559 |
3-4-5 | 19 (33.9%) | 39 | 56 | |||
Floor material | Land | 2 (50%) | 2 | 4 | 0.5 (0.81–4.24) | 0.595 |
Cement tile | 90 (37%) | 153 | 243 | |||
Water quality | Treated | 64 (33.2%) | 129 | 193 | 0.4 (0.20–0.850) | 0.012 |
Untreated | 28 (51.9%) | 26 | 54 | |||
Pets | Yes | 31 (37.8%) | 51 | 82 | 1.0 (0.600–1.79) | 0.898 |
No | 61 (37%) | 104 | 165 |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Osorio-Pulgarin, M.I.; Higuera, A.; Beltran-Álzate, J.C.; Sánchez-Jiménez, M.; Ramírez, J.D. Epidemiological and Molecular Characterization of Blastocystis Infection in Children Attending Daycare Centers in Medellín, Colombia. Biology 2021, 10, 669. https://doi.org/10.3390/biology10070669
Osorio-Pulgarin MI, Higuera A, Beltran-Álzate JC, Sánchez-Jiménez M, Ramírez JD. Epidemiological and Molecular Characterization of Blastocystis Infection in Children Attending Daycare Centers in Medellín, Colombia. Biology. 2021; 10(7):669. https://doi.org/10.3390/biology10070669
Chicago/Turabian StyleOsorio-Pulgarin, Maria I., Adriana Higuera, Juan C. Beltran-Álzate, Miryan Sánchez-Jiménez, and Juan David Ramírez. 2021. "Epidemiological and Molecular Characterization of Blastocystis Infection in Children Attending Daycare Centers in Medellín, Colombia" Biology 10, no. 7: 669. https://doi.org/10.3390/biology10070669
APA StyleOsorio-Pulgarin, M. I., Higuera, A., Beltran-Álzate, J. C., Sánchez-Jiménez, M., & Ramírez, J. D. (2021). Epidemiological and Molecular Characterization of Blastocystis Infection in Children Attending Daycare Centers in Medellín, Colombia. Biology, 10(7), 669. https://doi.org/10.3390/biology10070669