The penetration ability of 12 antimicrobial agents, including antibiotics and biocides, was determined against biofilms of B. cereus and P. fluorescens using a colony biofilm assay. The surfactants benzalkonium chloride (BAC) and cetyltrimethyl ammonium bromide (CTAB), and the antibiotics ciprofloxacin and streptomycin were of interest due to their distinct activities. Erythromycin and CTAB were retarded by the presence of biofilms, whereas ciprofloxacin and BAC were not. The removal and killing efficacies of these four agents was additionally evaluated against biofilms formed in microtiter plates. The most efficient biocide was CTAB for both bacterial biofilms. Ciprofloxacin was the best antibiotic although none of the selected antimicrobial agents led to total biofilm removal and/or killing. Comparative analysis of the results obtained with colony biofilms and microtiter plate biofilms show that although extracellular polymeric substances and the biofilm structure are considered a determining factor in biofilm resistance, the ability of an antimicrobial agent to penetrate a biofilm is not correlated with its killing or removal efficiency. Also, the results reinforce the role of an appropriate antimicrobial selection as a key step in the design of disinfection processes for biofilm control.Keywords: antimicrobial resistance; Bacillus cereus; biofilm control; Pseudomonas fluorescens; disinfection; diffusional limitations
IntroductionA biofilm is commonly defined as a microbial community with cells irreversibly attached to a substratum or attached to each other, and embedded in a matrix of extracellular polymeric substances (EPS) (Donlan & Costerton 2002). EPS protects bacteria from environ-mental adversities (Jefferson 2004). In all industries, especially in the food industry, the proliferation of microorganisms is very common even when manufacturers diligently follow all the necessary contingency plans (Araújo et al. 2011). The main objective of microbial control is to eliminate microorganisms or reduce their numbers to acceptable levels, as well as to prevent and control the formation of biological deposits attached to equipment surfaces (Maukonen et al. 2003). Currently, there is no control strategy capable of entirely eradicating biofilms (Simões et al. 2010). There is a need to find new strategies to manage antimicrobial resistance (Lee et al. 2013; Traba et al. 2013). 2 Resistance is the ability to withstand antimicrobial treatments. Russell (1999) and Chapman (2003) documented three types of resistance: intrinsic resistance, eg the Gram-negative lipopolysaccharide layer (McDonnell & Russell 1999); acquired resistance, eg manipulated resistance mediated by plasmids; and adaptive resistance, eg exposure to sub-lethal concentrations of an antimicrobial agent that selects for mutation, confer-ring resistance to that agent or others of the same type (cross-resistance). The way microorganisms develop resistance is not well understood. However, biofilm formation is a case of adaptive resistance and is cons...