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High throughput comparative assessment of biofilm formation of Candida glabrata on polystyrene material

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Abstract

Candida glabrata is the second most reported Candida species causing candidiasis next only to Candida albicans. Biofilm formation by C. glabrata accelerates pathogenicity, so understanding conditions for biofilm formation in relevant polystyrene polymers is essential. Determining optimum conditions for biofilm formation using one variable at a time (OVAT) is laborious and inaccurate. Therefore, mathematical modelling for design of experiments to optimize conditions for biofilm formation in Candida species was undertaken. In this study, a simple and robust statistical method, response surface methodology (RSM) was used to optimize independent variables like temperature, incubation period, media pH, shaker speed and inoculum density. Biofilm forming conditions were optimized and compared for two important local clinical isolates and one standard culture on 96 well polystyrene microtiter plates. Biofilm was quantified by different methods, like XTT for cell viability, crystal violet for biofilm, calcofluor white, and wet and dry weight measurements for cell mass. Quantification of cell viability and biofilm indicated heterogeneity among the three cultures. The results revealed an important finding that foetal bovine serum (FBS) does not significantly affect biofilm formation in vitro. This simple high throughput method for optimization and quantification of Candida biofilm has relevance for applications in rapid screening of anti-Candida biologics and therapeutic solutions while advancing control measures in polystyrene carriers in hospital settings.

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Acknowledgements

Financial support by DST, Govt. of India, grant no. SR/FT/LS-124/2012 to Bindu Sadanandan, principal investigator is acknowledged. Dr Beena Hemanth wishes to thank Dr Rameez Raja and Dr Indumati V A from M S Ramaiah Medical College and Teaching Hospital, Bengaluru for their help with the isolation of the cultures.

Funding

Financial support was provided by Department of Science and Technology (DST) Government of India. The authors have no conflict of interest. The funders have no role in the study design, data collection interpretation or the decision to submit the work for publication.

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Authors and Affiliations

  1. Department of Biotechnology, M S Ramaiah Institute of Technology, Bengaluru, India

    Bindu Sadanandan, Priya Ashrit, Lokesh Kyathsandra Nataraj, Amruta Puroshottam Jogalekar & Vijayalakshmi Vaniyamparambath

  2. Department of Plant Science, North Dakota State University, Fargo, ND, 58105, USA

    Kalidas Shetty

  3. Department of Microbiology, M S Ramaiah Medical College and Teaching Hospital, Bengaluru, India

    Beena Hemanth

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  1. Bindu Sadanandan
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Contributions

B.S., designed and carried out the experimental work. PA., A.P.J. and V.V., also carried out the experimental work. L., K. N., supported with the RSM design of experiments. K.S., was involved in the designing of the experiments, analyzing the results and manuscript review. B.H., provided the clinical isolates. B.S., P. A., and L.K.N., drafted the manuscript. All authors reviewed and approved the manuscript.

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Correspondence to Bindu Sadanandan.

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Additional information as noted in the text. This information is available via the Internet at http://www.springer.com/chemistry/journal/11814.

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Sadanandan, B., Ashrit, P., Nataraj, L.K. et al. High throughput comparative assessment of biofilm formation of Candida glabrata on polystyrene material. Korean J. Chem. Eng. 39, 1277–1286 (2022). https://doi.org/10.1007/s11814-021-1054-3

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  • DOI: https://doi.org/10.1007/s11814-021-1054-3

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