Abstract
The agar-cloning technique of Blastocystis hominis has been observed in both solid and semisolid agar using Iscove’s modified Dulbecco’s medium. In this study, Philippine isolates of B. hominis were grown by pour-plate method in semisolid agar using Locke’s solution. Inoculated plates contained 0.7% agar, 10% horse serum, and 0.1% sodium thioglycollate. Plates were incubated at 37°C in a microaerophilic jar for 7–10 days. Biconvex disk-shaped colonies were seen abound at the bottom half of the medium. Colonies growing at the agar–glass interface were flat and consisted of thin layers of cells. From these colonies, large amoeboid cells were frequently seen on the periphery, whereas smaller cells were concentrated at the core. Analysis of the SSU rDNA genetically established the identity of the clones to be B. hominis. This is the first report on agar cloning of Blastocystis in a compound medium.
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References
Clark CG, Diamond LS (2002) Methods for cultivation of luminal parasitic protists of clinical importance. Clin Microbiol Rev 15:329–341
Lanuza MD, Carbajal JA, Villar J, Borras R (1997) Description of an improved method for Blastocystis hominis culture and axenization. Parasitol Res 83:60–63
Leelayoova S, Rangsin R, Taamasri P, Naaglor T, Thathaisong U, Mungthin M (2004) Evidence of waterborne transmission of Blastocystis hominis. Am J Trop Med Hyg 70:658–662
Ng GC, Tan KSW (1999) Colony growth as a step towards axenization of Blastocystis isolates. Parasitol Res 85:678–679
Rivera WL, Tachibana H, Silva-Tahat MRA, Uemura H, Kanbara H (1996) Differentiation of Entamoeba histolytica and E. dispar DNA from cysts present in stool specimens by polymerase chain reaction: its field application in the Philippines. Parasitol Res 82:585–589
Sohail M, Fischer PR (2005) Blastocystis hominis and travelers. Travel Med Infect Dis 3:33–38
Stenzel DJ, Boreham PFL (1996) Blastocytis hominis revisited. Clin Microbiol Rev 9:563–584
Suresh K, Smith HV, Tan TC (2005) Viable Blastocystis cysts in Scottish and Malaysian sewage samples. Appl Environ Microbiol 71:5619–5620
Tan SW, Singh M, Thong KT, Ho LC, Moe KT, Chen XQ, Ng GC, Yap EH (1996a) Clonal growth of Blastocystis hominis in soft agar with sodium thioglycollate. Parasitol Res 82:737–739
Tan SW, Singh M, Yap EH, Ho LC, Moe KT, Howe J, Ng GC (1996b) Colony formation of Blastocystis hominis in soft agar. Parasitol Res 82:375–377
Tan KSW, Ng GC, Quek E, Howe J, Ramachandran NP, Yap EH, Singh M (2000) Blastocystis hominis: a simplified and high-efficiency method for clonal growth on solid agar. Exp Parasitol 96:9–15
Tan KSW, Howe J, Yap EH, Singh M (2001) Do Blastocystis hominis colony forms undergo programmed cell death? Parasitol Res 87:362–367
Tan KSW, Singh M, Yap EH (2002) Recent advances in Blastocystis hominis research: hot spots in terra incognita. Int J Parasitol 32:789–804
Yoshikawa H, Abe N, Iwasawa M, Kitano S, Nagano I, Wu Z, Takahashi Y (2000) Genomic analysis of Blastocystis hominis strains isolated from two long-term health care facilities. J Clin Microbiol 38:1324–1330
Yoshikawa H, Morimoto K, Nagashima M, Miyamoto N (2004) A survey of Blastocystis infection in anuran and urodele amphibians. Vet Parasitol 122:91–102
Zaman V, Howe J, Ng M (1997) Observations on the surface coat of Blastocystis hominis. Parasitol Res 83:731–733
Zaman V, Howe J, Ng M, Goh TK (1999) Scanning electron microscopy of the surface coat of Blastocystis hominis. Parasitol Res 85:974–976
Zierdt CH (1991) Blastocystis hominis—past and future. Clin Microbiol Rev 4:61–79
Acknowledgements
This work was supported by grants from the Natural Sciences Research Institute of the University of the Philippines (BIO-05-1-02) and the Department of Agriculture—Bureau of Agricultural Research to W.L.R.
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The experiments conducted in this study comply with the current laws of the Philippines.
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Valido, E.M., Rivera, W.L. Colony growth of Philippine isolates of Blastocystis hominis in simplified, soft agar medium. Parasitol Res 101, 213–217 (2007). https://doi.org/10.1007/s00436-006-0456-9
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DOI: https://doi.org/10.1007/s00436-006-0456-9