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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

Autofluorescence of different morphological forms of Blastocystis spp.
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Related In: Results  -  Collection


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Fig2: Autofluorescence of different morphological forms of Blastocystis spp.

Mentions: Autofluorescence was observed in VF, GF and AF and the CF (Fig. 2) of Blastocystis cells. The autofluorescence was greatest in the TRITC excitation/emission light spectra (557/576-nM) and appeared to be brightest in the central vacuole, with spots of light emission observed at the surface of the organism. The autofluorescence faded with time and appeared to be optimal in fresh VF forms, CF and least in AF. The autofluorescence was distracting but could be distinguished from the Blastofluor® stain in the VF and GF. The Blastofluor® stained the external cell membrane of VF, GF but not the AF (Fig. 3) and was brightest using the FITC (495/515-nm) light spectrum. The Blastofluor® stain could not be reliably differentiated from the strong autofluorescence seen in the CF. The Blastofluor® antibody stained the VF and GF of STs 1, 3 and 4 specimens but did not stain these forms in ST5 pig samples. DAPI staining of VF stained the nuclei on opposite poles of the central vacuole a dense bright blue.Fig. 2


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

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

Autofluorescence of different morphological forms of Blastocystis spp.
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig2: Autofluorescence of different morphological forms of Blastocystis spp.
Mentions: Autofluorescence was observed in VF, GF and AF and the CF (Fig. 2) of Blastocystis cells. The autofluorescence was greatest in the TRITC excitation/emission light spectra (557/576-nM) and appeared to be brightest in the central vacuole, with spots of light emission observed at the surface of the organism. The autofluorescence faded with time and appeared to be optimal in fresh VF forms, CF and least in AF. The autofluorescence was distracting but could be distinguished from the Blastofluor® stain in the VF and GF. The Blastofluor® stained the external cell membrane of VF, GF but not the AF (Fig. 3) and was brightest using the FITC (495/515-nm) light spectrum. The Blastofluor® stain could not be reliably differentiated from the strong autofluorescence seen in the CF. The Blastofluor® antibody stained the VF and GF of STs 1, 3 and 4 specimens but did not stain these forms in ST5 pig samples. DAPI staining of VF stained the nuclei on opposite poles of the central vacuole a dense bright blue.Fig. 2

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