Champion, AC and Houston, NK and Bradbury, RS and Reid, DWEC, Preliminary feasibility and modelling of a liquid matrix Dictyostelium discoideum virulence assay for Pseudomonas aeruginosa, British Journal of Biomedical Science, 73, (2) pp. 51-55. ISSN 0967-4845 (2016) [Refereed Article]
© 2016 British Journal of Biomedical Science
Objectives: To develop and determine the feasibility of using a liquid matrix adaptation of the Dictyostelium discoideum bacterial virulence assay by testing on well-characterised clinical and environmental isolates of Pseudomonas aeruginosa.
Materials and methods: Axenic AX2 D. discoideum were co-cultured with clinical and environmental isolates of P. aeruginosa in costar 24-well tissue culture plates for 24 h. A P. aeruginosa PAO1 positive control was tested in biological quintuplicate. Wells were then inspected using an inverted microscope and the degree of cytotoxic changes (sparse growth compared to control combined with rounding of cells and cytoplasmic shrinkage) on the D. discoideum cells was observed. A Klebsiella aerogenes negative control was included with each assay series.
Results: Sixty-five clinical and 20 environmental P. aeruginosa isolates were tested in the model. Cystic fibrosis respiratory isolates were found to be significantly (P < 0.05) less cytotoxic than P. aeruginosa from other sources. Limitations attached to the funding of this paper did not allow validation against previously employed models or animal models. Discussion: A liquid matrix D. discoideum model for the analysis of P. aeruginosa virulence in a eukaryotic host is feasible, but further validation of the model is required before it may be employed in routine setting.
|Item Type:||Refereed Article|
|Keywords:||Dictyostelium discoideum; Pseudomonas aeruginosa; virulence model|
|Research Division:||Medical and Health Sciences|
|Research Group:||Medical Microbiology|
|Research Field:||Medical Bacteriology|
|Objective Group:||Clinical Health (Organs, Diseases and Abnormal Conditions)|
|Objective Field:||Infectious Diseases|
|Author:||Champion, AC (Dr Alan Champion)|
|Author:||Houston, NK (Mrs Neralee Houston)|
|Web of Science® Times Cited:||3|
|Deposited By:||Medicine (Discipline)|
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