Axenic culture ofBlastocystis hominis in Iscove's modified Dulbecco's medium

Ho, L. C.; Singh, Mulkit; Suresh, G.; Ng, G. C.; Yap, E. H.

doi:10.1007/bf00932249

Cited by 71 publications

(64 citation statements)

References 8 publications

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“…Other studies utilized diphasic agar slant medium, which was useful for the culture of Blastocystis isolates from cattle, pigs, and chickens (2-4, 109, 110). Axenized cultures, that is, cultures of Blastocystis cells not associated with any other living organism, display luxuriant growth in a variety of media such as Iscove's modified Dulbecco's medium, minimal essential medium, or biphasic inspissated egg slant overlaid with Locke's solution (98,324). Cell densities of up to 2.5 ϫ 10 7 cells/ml can be attained for monophasic medium (98), while slightly higher densities of approximately 6.0 ϫ 10 7 cells/ml were reported for cells cultured in biphasic inspissated egg medium (324).…”

Section: Laboratory Culturementioning

confidence: 99%

“…Axenized cultures, that is, cultures of Blastocystis cells not associated with any other living organism, display luxuriant growth in a variety of media such as Iscove's modified Dulbecco's medium, minimal essential medium, or biphasic inspissated egg slant overlaid with Locke's solution (98,324). Cell densities of up to 2.5 ϫ 10 7 cells/ml can be attained for monophasic medium (98), while slightly higher densities of approximately 6.0 ϫ 10 7 cells/ml were reported for cells cultured in biphasic inspissated egg medium (324). The doubling time of axenic isolates can be variable, ranging from 6 to 23 h, depending on the isolate, study, and type of medium used (33,324).…”

Section: Laboratory Culturementioning

confidence: 99%

“…The doubling time of axenic isolates can be variable, ranging from 6 to 23 h, depending on the isolate, study, and type of medium used (33,324). Doubling times of approximately 50 h can be deduced from growth curves of Blastocystis isolates belonging to avian subtype 7 (98,169), and this may be due to the nonoptimal incubation temperature, as avian hosts, particularly chickens, generally have higher body temperatures than mammals. The colony growth of Blastocystis cells can be established in soft agar (Fig.…”

Section: Laboratory Culturementioning

confidence: 99%

“…It may be worthwhile to note that avian subtypes 6 and 7 may grow optimally at 40°C instead of 37°C, as is the case for the avian protozoan flagellate Histomonas meleagridis (84,279). Isolates belonging to subtype 7 have longer doubling times, about 50 h, when cultured at 37°C (98). In this case, these slow-growing subtypes may still be missed during in vitro culture expansion of stool samples, resulting in an underrepresentation of such subtypes in epidemiological surveys.…”

Section: Epidemiology and Prevalencementioning

confidence: 99%

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Extreme anoxia tolerance in embryos of the annual killifishAustrofundulus limnaeus: insights from a metabolomics analysis

Podrabsky

Lopez

Fan

et al. 2007

Journal of Experimental Biology

129

154

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SUMMARY The annual killifish Austrofundulus limnaeus survives in ephemeral pond habitats by producing drought-tolerant diapausing embryos. These embryos probably experience oxygen deprivation as part of their normal developmental environment. We assessed the anoxia tolerance of A. limnaeus embryos across the duration of embryonic development. Embryos develop a substantial tolerance to anoxia during early development, which peaks during diapause II. This extreme tolerance of anoxia is retained during the first 4 days of post-diapause II development and is then lost. Metabolism during anoxia appears to be supported mainly by production of lactate, with alanine and succinate production contributing to a lesser degree. Anoxic embryos also accumulate large quantities of γ-aminobutyrate (GABA), a potential protector of neural function. It appears that the suite of characters associated with normal development and entry into diapause II in this species prepares the embryos for long-term survival in anoxia even while the embryos are exposed to aerobic conditions. This is the first report of such extreme anoxia tolerance in a vertebrate embryo, and introduces a new model for the study of anoxia tolerance in vertebrates.

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Section: Laboratory Culturementioning

confidence: 99%

Section: Laboratory Culturementioning

confidence: 99%

Section: Laboratory Culturementioning

confidence: 99%

Section: Epidemiology and Prevalencementioning

confidence: 99%

See 2 more Smart Citations

Extreme anoxia tolerance in embryos of the annual killifishAustrofundulus limnaeus: insights from a metabolomics analysis

Podrabsky

Lopez

Fan

et al. 2007

Journal of Experimental Biology

129

154

View full text Add to dashboard Cite

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“…Blastocystis subtype 7 (previously known as Blastocystis hominis isolate B) was isolated from a local patient stool sample and axenized (Ho et al, 1993). Cells were cultured in Iscove's modified Dulbecco's medium (IMDM) containing 10 % inactive horse serum and incubated anaerobically at 37 uC in an Anaerojar (Oxoid).…”

Section: Methodsmentioning

confidence: 99%

Staurosporine-induced programmed cell death in Blastocystis occurs independently of caspases and cathepsins and is augmented by calpain inhibition

2010

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Previous studies have shown that the protozoan parasite Blastocystis exhibits apoptotic features with caspase-like activity upon exposure to a cytotoxic monoclonal antibody or the anti-parasitic drug metronidazole. The present study reports that staurosporine (STS), a common apoptosis inducer in mammalian cells, also induces cytoplasmic and nuclear features of apoptosis in Blastocystis, including cell shrinkage, phosphatidylserine (PS) externalization, maintenance of plasma membrane integrity, extensive cytoplasmic vacuolation, nuclear condensation and DNA fragmentation. STS-induced PS exposure and DNA fragmentation were abolished by the mitochondrial transition pore blocker cyclosporine A and significantly inhibited by the broad-range cysteine protease inhibitor iodoacetamide. Interestingly, the apoptosis phenotype was insensitive to inhibitors of caspases and cathepsins B and L, while calpain-specific inhibitors augmented the STS-induced apoptosis response. While the identities of the proteases responsible for STSinduced apoptosis warrant further investigation, these findings demonstrate that programmed cell death in Blastocystis is complex and regulated by multiple mediators. INTRODUCTIONProgrammed cell death (PCD) has long been recognized as an essential process to eliminate unwanted or damaged cells and thus ensure normal growth and development in multicellular organisms (Baehrecke, 2002;Hengartner, 2000). It was assumed that PCD arose with multicellular organisms (Vaux et al., 1994). However, considerable experimental evidence supports the existence of PCD in unicellular eukaryotes. These include non-parasitic organisms, such as yeast (Madeo et al., 2002), the free-living slime mould Dictyostelium discoideum (Arnoult et al., 2001;Cornillon et al., 1994), the free-living ciliate Tetrahymena thermophila (Christensen et al., 1998;Kobayashi & Endoh, 2005) and the dinoflagellate Peridinium gatunense (Vardi et al., 1999). In parasitic organisms, PCD has been described in the kinetoplastid trypanosomes (Ameisen et al., 1995;Welburn et al., 1996) and Leishmania (Arnoult et al., 2002;Bera et al., 2003;Zangger et al., 2002), the apicomplexan parasite Plasmodium Unicellular protozoan parasites cause a wide variety of human diseases. The current treatment of these infections is being challenged by an increasing incidence of drug resistance and a lack of effective vaccines (Croft et al., 2006;Fidock et al., 2008). The investigation of PCD pathways in these organisms might lead to the discovery of novel parasite control strategies (Alvarez et al., 2008;Deponte & Becker, 2004). However, despite the many morphological and biochemical studies of PCD in protozoan parasites, most of the homologues of mammalian molecules involved in cell death signalling are missing in the protozoa, and the molecular architecture of PCD in protozoan parasites therefore remains puzzling.Blastocystis is a unicellular protozoan parasite found in the intestines of humans and many animals (Tan, 2004(Tan, , 2008. It is transmitted through the fa...

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Blastocystis hominis

Windsor

2010

Topley &Amp; Wilson's Microbiology and Microbial Infections

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Axenic culture ofBlastocystis hominis in Iscove's modified Dulbecco's medium

Cited by 71 publications

References 8 publications

Extreme anoxia tolerance in embryos of the annual killifishAustrofundulus limnaeus: insights from a metabolomics analysis

Extreme anoxia tolerance in embryos of the annual killifishAustrofundulus limnaeus: insights from a metabolomics analysis

Staurosporine-induced programmed cell death in Blastocystis occurs independently of caspases and cathepsins and is augmented by calpain inhibition

Blastocystis hominis

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