title: The effect of in vitro growth conditions on the resistance of Acanthamoeba cysts.
abstract: Despite increasing concerns of direct pathogenicity and/or their role as hosts for other microorganisms there are currently no standard methods for the inactivation of amoebae that belong to the genus Acanthamoeba. Methods used to grow amoebae and produce cysts for these tests may be important as they can dramatically modify cyst susceptibility. We compared resistance of cysts produced from trophozoites grown in peptone-yeast extract-glucose broth or by feeding on HEp-2 cells and then encysted in Neff's medium. We observed that trophozoites grown using HEp-2 cells as a nutrient source produce cysts that are significantly more resistant to SDS and to most biocides tested, including heat. Increased resistance is likely due to a higher proportion of mature cysts presenting thicker cell walls as demonstrated using transmission electron microscopy. This was confirmed by calcofluor white staining demonstrating higher cellulose content in cysts produced from trophozoites grown using HEp-2 cells as a feeding source. These results demonstrate that not only methods used to produce cysts from trophozoites are critical, but that methods used to grow trophozoites before encystment should also be chosen carefully. This should be taken into account for the development of protocols to evaluate biocides and antimicrobials against amoebal cysts.
authors.affiliation: STERIS SA R&D, Fontenay-aux-Roses, France.
authors.forename: Celine
authors.initials: C
authors.lastname: Coulon
authors.forename: Nathalie
authors.initials: N
authors.lastname: Dechamps
authors.forename: Thierry
authors.initials: T
authors.lastname: Meylheuc
authors.forename: Anne
authors.initials: A
authors.lastname: Collignon
authors.forename: Gerald
authors.initials: G
authors.lastname: McDonnell
authors.forename: Vincent
authors.initials: V
authors.lastname: Thomas
mesh_terms: D000048:Acanthamoeba / Q000187:drug effects* / Q000254:growth & development* / Q000648:ultrastructure; D000818:Animals; D000890:Anti-Infective Agents / Q000494:pharmacology*; D002460:Cell Line; D004202:Disinfectants / Q000494:pharmacology*; D004351:Drug Resistance; D006801:Humans; D008855:Microscopy, Electron, Scanning; D021261:Parasitic Sensitivity Tests; D053058:Trophozoites / Q000187:drug effects / Q000254:growth & development / Q000648:ultrastructure
chemical_list: D000890:Anti-Infective Agents; D004202:Disinfectants