Abstract
Blastocystis sp. is known to be the most commonly found intestinal protozoan parasite in human fecal surveys and has been incriminated to cause diarrhea and abdominal bloating. Binary fission has been widely accepted as the plausible mode of reproduction for this parasite. The present study demonstrates that subjecting the parasites in vitro to higher temperature shows the proliferation of parasite numbers in cultures. Transmission electron microscopy was used to compare the morphology of Blastocystis sp. subtype 3 isolated from a dengue patient having high fever (in vivo thermal stress) and Blastocystis sp. 3 maintained at 41 °C (in vitro thermal stress) and 37 °C (control). Fluorescence stains like acridine orange (AO) and 4′,6′-diamino-2-phenylindole (DAPI) were used to demonstrate the viability and nuclear content of the parasite for both the in vitro and in vivo thermal stress groups of parasites. Blastocystis sp. at 37 °C was found to be mostly vacuolar whereas the in vitro thermal stressed isolates at 41 °C were granular with electron dense material seen to protect the granules within the central body. Parasites of the in vivo thermal stressed group showed similar ultrastructure as the in vitro ones. AO and DAPI staining provided evidence that these granules are viable which develop into progenies of Blastocystis sp. These granular forms were then observed to rupture and release progenies from the mother cells whilst the peripheral cytoplasmic walls were seen to degrade. Upon exposure to high temperature both in vitro and in vivo, Blastocystis sp. in cultures show higher number of granular forms seen to be protected by the electron dense material within the central body possibly acting as a protective mechanism. This is possibly to ensure the ability to survive for the granules to be developed as viable progenies for release into the host system.
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Acknowledgements
We would like to thank our lab colleagues and all the staffs of Department of Parasitology, University of Malaya. We also would like to thank the staffs of Electron Microscopy Unit, Faculty of Medicine, University of Malaya for the guidance on transmission electron microscopy work.
Funding
Funding for this study was provided by the Fundamental Research Grant Scheme by the Ministry of Higher Education (FRGS) (FP015-2017A) and University Malaya Postgraduate Research Fund (PPP) (PG141-2016A). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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GT performed the experiments. GT, SKG and SB were involved in the intellectual planning of the experiment. GT, and SKG analyzed the results, wrote the paper and approved the final manuscript. All authors read and approved the final version of the manuscript.
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The collection and storage of samples for research purposes was approved by the University of Malaya Medical Ethics committee (MECID.NO: 20151-984).
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Thergarajan, G., Govind, S.K. & Bhassu, S. In vitro and in vivo thermal stress induces proliferation of Blastocystis sp.. Parasitol Res 117, 177–187 (2018). https://doi.org/10.1007/s00436-017-5688-3
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DOI: https://doi.org/10.1007/s00436-017-5688-3