A microplate‐based Response Surface Methodology model for growth optimization and biofilm formation on polystyrene polymeric material in a Candida albicans and Escherichia coli co‐culture

Abstract

Biofilm formation is one of the crucial factors for virulence in Candida albicans, Escherichia coli and their polymicrobial biofilm formed on medical prosthetics. In this study, high biofilm forming C. albicans M207 from a case of invasive candidiasis and a standard strain of bacterial culture E. coli ATCC 39936 were used for initial screening and media optimization using One Variable at a time (OVAT) and Response Surface Methodology (RSM) models, respectively, in polystyrene microtiter plates for in vitro biofilm studies. Sucrose and peptone significantly affected biofilm formation when screened by OVAT. Media optimization by Central Composite Design (CCD) showed that pH significantly affected biofilm formation for all the cultures which was further validated at critical parameters. Biofilm was quantified by growth OD measurements, MTT and CV assays in the presence and absence of FBS precoating and compared with standard Trypticase Soy Broth (TSB) media which did not show any significant difference. This high throughput polystyrene microtiter plate assay for preliminary screening of nutritional components using OVAT and media optimization using RSM model is being reported for the first time for biofilm formation by C. albicans and E. coli in a single and dual species environment. This study is essential for functional analysis of in‐vitro growth and biofilm formation by these two pathogenic strains on polymeric hospital equipments and to develop suitable control measures to inhibit biofilm formation by C. albicans and E. coli single species and their co‐culture.

Response Surface Methodology Application for Bacteriophage–Antibiotic Antibiofilm Activity Optimization

“…The application of RSM in optimizing C. albicans growth and biofilm formation offers a promising approach for future research in screening therapeutic interventions and advancing the control of biofilm-related challenges. This novel method significantly enhances our understanding of pathogenic biofilm interactions and paves the way for effective biofilm management strategies [ 34 , 35 ].…”

Cerium oxide particles: coating with charged polysaccharides for limiting the aggregation state in biological media and potential application for antibiotic delivery

“…A biofilm of E. coli was prepared on a polystyrene plate by using the protocol developed by Ashrit et al 40 The bacterial viability was investigated via an orange acridine test coupled with fluorescence microscopy observation. 41…”

Silkworm model of biofilm formation: In vivo evaluation of antimicrobial tolerance of a cross-kingdom dual-species (Escherichia coli and Candida albicans) biofilm on catheter material

“…using microplates [45,46]. Based on these techniques, a method has been established to measure polymicrobial biofilm formation by C. albicans and E. coli [47]. Moreover, the method can be used to identify samples that inhibit the polymicrobial biofilm formation of C. albicans and E. coli [48].…”