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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

Morphological forms of Blastocystis sp. in xenic culture stained with acridine orange
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Related In: Results  -  Collection


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Fig1: Morphological forms of Blastocystis sp. in xenic culture stained with acridine orange

Mentions: Studies utilising light microscopy, transmission (TEM), scanning (SEM) and freeze-etch (FE-EM) electron microscopy have described multiple forms of the Blastocystis organism, including vacuolated (VF), granular (GF), amoebic (AF) and cystic (CF) forms (Fig. 1). The relationship of these different forms to each other is unclear (Tan 2008), although it is certain that the robust cystic form transmits infection (Moe et al. 1997). Microscopic images have often been obtained from attenuated or axenic cultures. These elegant studies have been useful in describing the intricate ultrastructure and surface morphology of the various forms of Blastocystis spp., but their limitation is that they capture still images of a dead organism separated from the usual microbial environment. In this study, we employed deconvolutional microscopy of xenic cultures of living Blastocystis sp. to obtain time-lapse and three-dimensional images of the Blastocystis organisms. This microscope also has the capability to record fluorescence in various light spectra facilitating utilisation of Blastocystis-specific fluorescent antibodies and fluorescent double-stranded deoxyribonucleic acid (DNA) stain, 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, Australia).Fig. 1


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

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

Morphological forms of Blastocystis sp. in xenic culture stained with acridine orange
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig1: Morphological forms of Blastocystis sp. in xenic culture stained with acridine orange
Mentions: Studies utilising light microscopy, transmission (TEM), scanning (SEM) and freeze-etch (FE-EM) electron microscopy have described multiple forms of the Blastocystis organism, including vacuolated (VF), granular (GF), amoebic (AF) and cystic (CF) forms (Fig. 1). The relationship of these different forms to each other is unclear (Tan 2008), although it is certain that the robust cystic form transmits infection (Moe et al. 1997). Microscopic images have often been obtained from attenuated or axenic cultures. These elegant studies have been useful in describing the intricate ultrastructure and surface morphology of the various forms of Blastocystis spp., but their limitation is that they capture still images of a dead organism separated from the usual microbial environment. In this study, we employed deconvolutional microscopy of xenic cultures of living Blastocystis sp. to obtain time-lapse and three-dimensional images of the Blastocystis organisms. This microscope also has the capability to record fluorescence in various light spectra facilitating utilisation of Blastocystis-specific fluorescent antibodies and fluorescent double-stranded deoxyribonucleic acid (DNA) stain, 4′,6-diamidino-2-phenylindole (DAPI) (Sigma-Aldrich, Australia).Fig. 1

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