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Features of Blastocystis spp. in xenic culture revealed by deconvolutional microscopy.

Nagel R, Gray C, Bielefeldt-Ohmann H, Traub RJ - Parasitol. Res. (2015)

Bottom Line: Surface pores of 1 μm in diameter were observed cyclically opening and closing over 24 h.Vacuolated forms extruded a viscous material from a single surface point with coincident deflation that may demonstrate osmoregulation.Tear-shaped granules were observed exiting from the surface of an amoebic form, but their origin and identity remain unknown.

View Article: PubMed Central - PubMed

Affiliation: School of Veterinary Science, University of Queensland, Gatton, Queensland, Australia, robyn@tgclinic.com.au.

ABSTRACT
Blastocystis spp. are common human enteric parasites with complex morphology and have been reported to cause irritable bowel syndrome (IBS). Deconvolutional microscopy with time-lapse imaging and fluorescent spectroscopy of xenic cultures of Blastocystis spp. from stool samples of IBS patients and from asymptomatic, healthy pigs allowed observations of living organisms in their natural microbial environment. Blastocystis organisms of the vacuolated, granular, amoebic and cystic forms were observed to autofluorescence in the 557/576 emission spectra. Autofluorescence could be distinguished from fluorescein-conjugated Blastocystis-specific antibody labelling in vacuolated and granular forms. This antibody labelled Blastocystis subtypes 1, 3 and 4 but not 5. Surface pores of 1 μm in diameter were observed cyclically opening and closing over 24 h. Vacuolated forms extruded a viscous material from a single surface point with coincident deflation that may demonstrate osmoregulation. Tear-shaped granules were observed exiting from the surface of an amoebic form, but their origin and identity remain unknown.

No MeSH data available.


Related in: MedlinePlus

Xenic culture of Blastocystis granular forms, time-lapse images over 24 h
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Fig5: Xenic culture of Blastocystis granular forms, time-lapse images over 24 h

Mentions: Detailed views of Blastocystis organisms were obtained, but apart from observing binary fission developing in vacuolated and granular forms, we did not observe changes from one morphological form to another. It was difficult to keep the Blastocystis organisms in the field of view despite using the cell tracking facility of the microscope. The Blastocystis cells were fragile and would often either rupture (Fig. 4) or drift away out of the field of view over the 24 h of recording, preventing observation of progressive cell changes past a certain point (Fig. 5). One GF, observed over 6 h in total, that appeared stationary on the slide displayed a large central granule moving rapidly within the GF (Fig. 6, Supplementary video file 1). Another time-lapse study of a granular AF showed the form enlarging and losing granularity on the surface with subsequent expulsion of small tear-shaped granules. These granules appeared to arise from the surface of the organism or track along fissures to the surface (Fig. 7). These granules varied in size from 1 to 3 μm and appeared to have a central dark spot surrounded by clear cytoplasm with a thick dark surface membrane. The release of the granules from the surface was best appreciated in the video images (Supplementary video file 2). Granules that appear to have tracked along fissures are identified as an area of interest in Fig. 7.Fig. 4


Features of Blastocystis spp. in xenic culture revealed by deconvolutional microscopy.

Nagel R, Gray C, Bielefeldt-Ohmann H, Traub RJ - Parasitol. Res. (2015)

Xenic culture of Blastocystis granular forms, time-lapse images over 24 h
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4537705&req=5

Fig5: Xenic culture of Blastocystis granular forms, time-lapse images over 24 h
Mentions: Detailed views of Blastocystis organisms were obtained, but apart from observing binary fission developing in vacuolated and granular forms, we did not observe changes from one morphological form to another. It was difficult to keep the Blastocystis organisms in the field of view despite using the cell tracking facility of the microscope. The Blastocystis cells were fragile and would often either rupture (Fig. 4) or drift away out of the field of view over the 24 h of recording, preventing observation of progressive cell changes past a certain point (Fig. 5). One GF, observed over 6 h in total, that appeared stationary on the slide displayed a large central granule moving rapidly within the GF (Fig. 6, Supplementary video file 1). Another time-lapse study of a granular AF showed the form enlarging and losing granularity on the surface with subsequent expulsion of small tear-shaped granules. These granules appeared to arise from the surface of the organism or track along fissures to the surface (Fig. 7). These granules varied in size from 1 to 3 μm and appeared to have a central dark spot surrounded by clear cytoplasm with a thick dark surface membrane. The release of the granules from the surface was best appreciated in the video images (Supplementary video file 2). Granules that appear to have tracked along fissures are identified as an area of interest in Fig. 7.Fig. 4

Bottom Line: Surface pores of 1 μm in diameter were observed cyclically opening and closing over 24 h.Vacuolated forms extruded a viscous material from a single surface point with coincident deflation that may demonstrate osmoregulation.Tear-shaped granules were observed exiting from the surface of an amoebic form, but their origin and identity remain unknown.

View Article: PubMed Central - PubMed

Affiliation: School of Veterinary Science, University of Queensland, Gatton, Queensland, Australia, robyn@tgclinic.com.au.

ABSTRACT
Blastocystis spp. are common human enteric parasites with complex morphology and have been reported to cause irritable bowel syndrome (IBS). Deconvolutional microscopy with time-lapse imaging and fluorescent spectroscopy of xenic cultures of Blastocystis spp. from stool samples of IBS patients and from asymptomatic, healthy pigs allowed observations of living organisms in their natural microbial environment. Blastocystis organisms of the vacuolated, granular, amoebic and cystic forms were observed to autofluorescence in the 557/576 emission spectra. Autofluorescence could be distinguished from fluorescein-conjugated Blastocystis-specific antibody labelling in vacuolated and granular forms. This antibody labelled Blastocystis subtypes 1, 3 and 4 but not 5. Surface pores of 1 μm in diameter were observed cyclically opening and closing over 24 h. Vacuolated forms extruded a viscous material from a single surface point with coincident deflation that may demonstrate osmoregulation. Tear-shaped granules were observed exiting from the surface of an amoebic form, but their origin and identity remain unknown.

No MeSH data available.


Related in: MedlinePlus