Isolation and Morphologic Differentiation of Blastocystis from Insect Vectors Using Staining Media
Authors
School of Medical Laboratory Science, The Manila Times College of Subic (Philippines)
School of Medical Laboratory Science, The Manila Times College of Subic (Philippines)
School of Medical Laboratory Science, The Manila Times College of Subic (Philippines)
School of Medical Laboratory Science, The Manila Times College of Subic (Philippines)
Article Information
DOI: 10.51244/IJRSI.2025.12110145
Subject Category: Parasitology - Health Sciences
Volume/Issue: 12/11 | Page No: 1635-1643
Publication Timeline
Submitted: 2025-11-27
Accepted: 2025-12-02
Published: 2025-12-20
Abstract
The complexity of Blastocystis morphology, its presence in various insect vectors, and the need for accurate detection methods warrant further research. This comparative, cross-sectional study aimed to detect Blastocystis species in domestic insect vectors, including Periplaneta americana (cockroach), Musca domestica (housefly), and Drosophila melanogaster (fruit fly). The study also evaluated the effectiveness of various staining techniques, namely Acid Fast staining, Gram staining, Methylene blue wet mount, and Direct smear using Lugol's solution, for differential enumeration and morphologic assessment of Blastocystis cysts. The results showed that Blastocystis cysts were recovered from Periplaneta americana and Musca domestica. Notably, Methylene blue and Lugol's Iodine in Direct smear yielded higher morphologic scores, suggesting that these staining methods may be more effective for visualizing Blastocystis cysts in insect vectors.
Keywords
Blastocystis; insect vectors; staining media; external washings; intestinal contents.
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References
1. Tan KS (2008). New insights on classification, identification, and clinical relevance of Blastocystis spp. Clin Microbiol Rev. 2008 Oct;21(4):639-65. doi: 10.1128/CMR.00022-08. PMID: 18854485; PMCID: PMC2570156. [Google Scholar] [Crossref]
2. Stenzel DJ, Boreham PF (1996). Blastocystis hominis revisited. Clin Microbiol Rev. 1996 Oct;9(4):563-84. doi: 10.1128/CMR.9.4.563. PMID: 8894352; PMCID: PMC172910. [Google Scholar] [Crossref]
3. Leelayoova, S., et al. (2008). Genetic diversity of Blastocystis subtype 3 in human isolates from Thailand. Infection, Genetics and Evolution, 8(2), 161-166. [Google Scholar] [Crossref]
4. Park, S. K., et al. (2012). Prevalence and subtype distribution of Blastocystis in Korean patients with irritable bowel syndrome. Korean Journal of Parasitology, 50(2), 139-144. [Google Scholar] [Crossref]
5. Greige S, El Safadi D, et al (2018). Prevalence and subtype distribution of Blastocystis sp. isolates from poultry in Lebanon and evidence of zoonotic potential. Parasit Vectors. 2018 Jul 4;11(1):389. doi: 10.1186/s13071-018-2975-5. PMID: 29973261; PMCID: PMC6030734. [Google Scholar] [Crossref]
6. Graczyk, T. K., et al. (2005). Filth flies are transport hosts of Cryptosporidium parvum. Emerging Infectious Diseases, 11(5), 748-751. [Google Scholar] [Crossref]
7. Jones, M. S., et al. (2014). Blastocystis subtypes in symptomatic and asymptomatic family members and pets. Journal of Clinical Microbiology, 52(10), 3467-3473. [Google Scholar] [Crossref]
8. Roberts, T., et al. (2014). Blastocystis infection and subtype distribution in symptomatic and asymptomatic individuals. Journal of Clinical Microbiology, 52(10), 3461-3466. [Google Scholar] [Crossref]
9. Stensvold, C. R., et al. (2012). Detecting and identifying Blastocystis subtypes using PCR and sequencing. Methods in Molecular Biology, 862, 171-184. [Google Scholar] [Crossref]
10. Termmathurapoj, S., et al. (2004). Comparative evaluation of direct fecal smear and formalin-ether concentration techniques for diagnosis of Blastocystis hominis. Southeast Asian Journal of Tropical Medicine and Public Health, 35(2), 267-271. [Google Scholar] [Crossref]
11. Eymael, D., et al. (2008). Standardization of Blastocystis hominis diagnosis using different staining methods. Journal of Clinical Microbiology, 46(10), 3467-3470. [Google Scholar] [Crossref]
12. Bergamo do Bomfim, T. C., & Machado do Couto, M. C. (2013). Morphological diagnosis and occurrence of Blastocystis stained with Giemsa in domestic bird species. Revista Brasileira de Parasitologia Veterinária, 22(2), 249-254. [Google Scholar] [Crossref]
13. Khalifa, A. M., El-Sheikh, H. F., & El-Rigalaty, A. A. (1999). Evaluation of different staining methods for the detection of Blastocystis hominis. Journal of the Egyptian Society of Parasitology, 29(3), 741-754. [Google Scholar] [Crossref]
14. Rambo, W. R., et al. (2017). Cockroaches as vectors of pathogens in hospitals. Journal of Infection and Public Health, 10(5), 531-536. [Google Scholar] [Crossref]
15. Menasria, T., Moussa, A., & Bousseboua, H. (2014). Intestinal parasites in cockroaches in hospitals of Batna city, Algeria. Journal of Medical Entomology, 51(4), 853-858. [Google Scholar] [Crossref]
16. Graczyk, T. D., Knight, R., & Tam, E. W. (2001). Mechanical transmission of Cryptosporidium parvum oocysts by flies. American Journal of Tropical Medicine and Hygiene, 65(4), 309-314. [Google Scholar] [Crossref]
17. Alzahrani, S. M., & Mounkaila, S. (2019). Parasites of medical importance in flies: A review. Journal of Medical Entomology, 56(4), 831-844. [Google Scholar] [Crossref]
18. Graczyk, T. K., Knight, R., & Tamang, L. (2005). Mechanical transmission of human protozoan parasites by insects. Clinical Microbiology Reviews, 18(1), 128-132. [Google Scholar] [Crossref]
19. Fayer, R., & Lewis, E. J. (1998). Cockroaches as vectors of parasites. Journal of Medical Entomology, 35(5), 669-675. [Google Scholar] [Crossref]
20. Sia Su, GL, et al, (2018). Parasitic Infestation in Cockroaches (Periplaneta americana) Obtained in Selected Areas of Metro Manila. International Journal of TROPICAL DISEASE & Health, ISSN: 2278-1005,Vol.: 13, Issue.: 4 [Google Scholar] [Crossref]
21. Chinchilla, M., & Reyes, L. (1998). Toxoplasma gondii in cockroaches. Journal of Parasitology, 84(3), 537-539. [Google Scholar] [Crossref]
22. Levine, O. S., & Levine, M. M. (1991). Houseflies as vectors of diarrheal diseases. Journal of Medical Entomology, 28(4), 491-496. [Google Scholar] [Crossref]
23. Echeverria, P., & Taylor, D. N. (1991). Houseflies and the transmission of Shigella spp. Journal of Medical Entomology, 28(4), 497-502. [Google Scholar] [Crossref]
24. Pawlowski, Z. S. (1990). Taeniasis and cysticercosis. In K. S. Warren & A. A. F. Mahmoud (Eds.), Tropical and geographical medicine (pp. 655-665). McGraw-Hill. [Google Scholar] [Crossref]
25. Tan, K. S. (2008). Blastocystis in humans and animals: new insights using modern methodologies. Veterinary Parasitology, 151(2-4), 247-256. [Google Scholar] [Crossref]
26. Johnson, E. H., & Windsor, J. J. (2006). Dientamoeba fragilis: a neglected human pathogen. Journal of Infection, 53(2), 75-82. [Google Scholar] [Crossref]
27. Alfellani, M. A., et al. (2013). Genetic diversity of Blastocystis in animals and humans.Infection, Genetics and Evolution, 19, 232-239 [Google Scholar] [Crossref]
28. Ramsay, M., Reithinger, R., & Curtis, C. (2010). Blastocystis: a bug with a complex lifestyle. Trends in Parasitology, 26(11), 538-545. [Google Scholar] [Crossref]
29. Stensvold, C. R., Alfellani, M., & Clark, C. G. (2012). Blastocystis: to subtype or not to subtype? Journal of Clinical Microbiology, 50(11), 3564-3565. [Google Scholar] [Crossref]