title: Differences in isoenzyme patterns of axenically and monoxenically grown Acanthamoeba and Hartmannella.
abstract: Axenically and monoxenically grown Acanthamoeba castellanii, Acanthamoeba polyphaga and different isolates of Hartmannella vermiformis strains were examined by polyacrylamide isoelectric focusing in the pH range 3-10. Isoenzyme patterns of acid phosphatase (AP), propionyl esterase (PE), malate dehydrogenase (MDH), alcohol dehydrogenase (ADH), glucose phosphate isomerase (GPI) and phosphoglucomutase (PGM) were compared. Zymograms were used to reveal differences in typical isoenzyme patterns between axenically and monoxenically grown amoebae and to compare axenically grown A. castellanii, A. polyphaga and H. vermiformis. Comparison of zymograms for AP, PE and MDH between axenically grown Acanthamoeba and Hartmannella strains revealed different isoenzyme patterns. Acanthamoeba showed strong bands for ADH and extremely weak bands for GPI and PGM, while Hartmannella lacked ADH but possessed bands for GPI and PGM. Comparison of zymograms from axenically and monoxenically grown amoebae revealed a lower intensity and even lack of typical isoenzyme bands in lysates from monoxenic cultures. The observed changes in typical isoenzyme patterns induced by the bacterial substrate can influence the correct isoenzymatic typing of different strains in clinical and phylogenetic studies.
authors.affiliation: Department of Microbiology, Faculty of Science, University of Nijmegen, The Netherlands.
authors.forename: P H
authors.initials: PH
authors.lastname: Weekers
authors.forename: J F
authors.initials: JF
authors.lastname: De Jonckheere
mesh_terms: D000048:Acanthamoeba / Q000254:growth & development* / Q000378:metabolism*; D000135:Acid Phosphatase / Q000032:analysis / Q000302:isolation & purification; D000426:Alcohol Dehydrogenase / Q000032:analysis / Q000302:isolation & purification; D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000032:analysis / Q000302:isolation & purification; D018920:Coculture Techniques; D003470:Culture Media / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005956:Glucose-6-Phosphate Isomerase / Q000032:analysis / Q000302:isolation & purification; D006249:Hartmannella / Q000254:growth & development* / Q000378:metabolism*; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000032:analysis* / Q000302:isolation & purification*; D008291:Malate Dehydrogenase / Q000032:analysis / Q000302:isolation & purification; D010733:Phosphoglucomutase / Q000032:analysis / Q000302:isolation & purification; D011549:Pseudomonas / Q000378:metabolism
chemical_list: D003470:Culture Media; D007527:Isoenzymes; D000426:Alcohol Dehydrogenase; D008291:Malate Dehydrogenase; D002265:Carboxylic Ester Hydrolases; C034216:propionyl esterase; D000135:Acid Phosphatase; D005956:Glucose-6-Phosphate Isomerase; D010733:Phosphoglucomutase