Category Archives: Biofilm

Research – Behavior of foodborne pathogens, Listeria monocytogenes and Staphylococcus aureus, in mixed-species biofilm exposed to biocides


In nature and man-made environments, microorganisms reside in mixed-species biofilm where behavior is modified compared to the single-species biofilms. Pathogenic microorganisms may be protected against adverse treatments in mixed-species biofilms leading to health risk for humans. Here, we developed two mixed-five-species biofilms that included the foodborne pathogens Listeria monocytogenes or Staphylococcus aureus, respectively. The five species, including the pathogen, were isolated from a single food-processing environmental sample thus mimicking the environmental community. In mature mixed five-species biofilms on stainless steel, the two pathogens remained at a constant level of ∼105 CFU/cm2 The mixed-five-species biofilms as well as the pathogens in mono-species biofilms were exposed to biocides to determine any pathogen-protective effect of the mixed biofilm. Both pathogens and their associate microbial communities were reduced by peracetic acid treatments. S. aureus decreased 4.6 log cycles in mono-species biofilm, but the pathogen was protected in the five-species biofilm and decreased only 1.1 log cycles. Sessile cells of L. monocytogenes were affected equally as a mono-biofilm or as a member in the mixed-species biofilm; decreasing by three log cycles when exposed to 0.0375 % peracetic acid. When the pathogen was exchanged in each associate microbial community, S. aureus was eradicated while there was no significant effect of the biocide on L. monocytogenes or the mixed community. This indicates that particular members or associations in the community offered the protective effect. Further studies are needed to clarify the mechanisms of biocide protection, and the species playing the protective role in microbial communities of biofilms. Importance: This study demonstrates that foodborne pathogens can be established in mixed species biofilms and that this can protect them from biocide action. The protection is not due to specific characteristics of the pathogen, here S. aureus and L. monocytogenes, but likely caused by specific members or associations in the mixed species biofilm. Biocide treatment and resistance is a challenge for many industries and biocide efficacy should be tested on microorganisms growing in biofilms, preferably mixed systems, mimicking the application environment.

Research – Cinnamaldehyde disrupts biofilm formation and swarming motility of Pseudomonas aeruginosa

Microbiology Research

Bacterial biofilms can cause serious health care complications associated with increased morbidity and mortality. There is an urge to discover and develop new biofilm inhibitors from natural products or by modifying natural compounds or understanding the modes of action of existing compounds. Cinnamaldehyde (CAD), one of the major components of cinnamon oil, has been demonstrated to act as an antimicrobial agent against a number of Gram-negative and Gram-positive pathogens, including Pseudomonas aeruginosa, Helicobacter pylori and Listeria monocytogenes. Despite the mechanism of action of CAD against the model organism P. aeruginosa being undefined, based on its antimicrobial properties, we hypothesized that it may disrupt preformed biofilms of P. aeruginosa. The minimum inhibitory concentration (MIC) of CAD for planktonic P. aeruginosa was determined to be 11.8 mM. Membrane depolarization assays demonstrated disruption of the transmembrane potential of P. aeruginosa. CAD at 5.9 mM (0.5 MIC) disrupted preformed biofilms by 75.6 % and 3 mM CAD (0.25 MIC) reduced the intracellular concentrations of the secondary messenger, bis-(3′–5′)-cyclic dimeric guanosine monophosphate (c-di-GMP), which controls P. aeruginosa biofilm formation. The swarming motility of P. aeruginosa was also reduced by CAD in a concentration-dependent manner. Collectively, these findings show that sub-MICs of CAD can disrupt biofilms and other surface colonization phenotypes through the modulation of intracellular signalling processes.

Research – Insights into Bacterial Milk Spoilage with Particular Emphasis on the Roles of Heat-Stable Enzymes, Biofilms, and Quorum Sensing

Journal of Food Protection Raw Milk Food Safety KSW Foodworld

Milk spoilage caused by psychrotrophic bacteria and their heat-stable enzymes is a serious challenge for the dairy industry. In many studies, spoilage has been explored based on the simplistic view of undesirable enzymes produced by planktonic cells. Recently, biofilms and quorum sensing (QS) have been suggested as important factors in the deterioration of milk, which opens new avenues for investigation of the processes and challenges. Production and heat stability of enzymes are enhanced in biofilms, mainly because of inherent differences in physiological states and protective shielding by extracellular polymeric substances. QS plays a key role in modulating expression of hydrolytic enzymes and biofilm formation. To date, few studies have been conducted to investigate the complex interplays of enzyme production, biofilm formation, and QS. This review provides novel insights into milk spoilage with particular emphasis on the roles of biofilms and QS and summarizes potential effective strategies for controlling the spoilage of milk.

Research – A Comparative Study of the Susceptibility of Listeria Species to Sanitizer Treatments When Grown under Planktonic and Biofilm Conditions

Journal of Food Protection

Listeria species are ubiquitous in nature and can adapt to survive in a variety of niches, including food processing environments. Listeria species that colonize these environments may also have the potential to persist. Food safety strategies designed to manage these niches include regular cleaning and disinfection with proven sanitizers containing biocide-active compounds. Typically, these sanitizers are effective against bacteria growing under planktonic conditions, but their efficacy may be compromised when bacteria are contained in biofilms. The susceptibility of persistent Listeria isolates, i.e., those capable of forming biofilms, to a selection of sanitizers was investigated. A quaternary ammonium compound–based sanitizer was the biocide most effective against planktonic bacteria, with a MIC of 0.0015 to 0.006%. In contrast, ethanol-based sanitizers were the least effective. Although, no triclosan tolerance was observed for planktonic Listeria isolates, triclosan was the only biocide that resulted in a significant biomass reduction. Differences between Listeria species were observed; L. monocytogenes and L. welshimeri biofilms were more tolerant to quaternary ammonium compound–based sanitizers than were L. innocua biofilms. These findings extend our understanding of the application of commonly used sanitizers in the food industry and the efficacy of these sanitizers against Listeria species and their associated biofilms.

Research – Staphylococcus aureus gene involved in virulence, biofilm formation and resistance to certain antibiotics: Study

Outbreak News Today Staph

An Institut Pasteur-CNRS research team has characterized a Staphylococcus aureus gene involved in virulence, biofilm formation and resistance to certain antibiotics. These results open up new avenues for understanding the control of S. aureus virulence mechanisms. This work was recently published in the journal PLoS Pathogens.

Staphylococcus aureus is part of the natural skin flora, preferentially colonizing external mucosa in 30 to 50% of the population, healthy carriers who develop no symptoms. But it is also a major human pathogen, causing diseases ranging from skin lesions (boils, impetigo, etc.) to endocarditis, acute pneumonia, osteomyelitis or sepsis. It is the leading Gram-positive bacterium responsible for nosocomial infections (hospital acquired infections). The most dangerous strains are those that display resistance to multiple antibiotics. This is the case of MRSA, resistant to Methicillin, widespread in hospitals and posing a major public health concern.

Research – Cross-Contamination and Biofilm Formation by Salmonella enterica Serovar Enteritidis on Various Cutting Boards

Mary Ann Liebert

Cross-contamination is one of the main factors related to foodborne outbreaks. This study aimed to analyze the cross-contamination process of Salmonella enterica serovar Enteritidis from poultry to cucumbers, on various cutting board surfaces (plastic, wood, and glass) before and after washing and in the presence and absence of biofilm. Thus, 10 strains of Salmonella Enteritidis were used to test cross-contamination from poultry to the cutting boards and from thereon to cucumbers. Moreover, these strains were evaluated as to their capacity to form biofilm on hydrophobic (wood and plastic) and hydrophilic materials (glass). We recovered the 10 isolates from all unwashed boards and from all cucumbers that had contacted them. After washing, the recovery ranged from 10% to 100%, depending on the board material. In the presence of biofilm, the recovery of salmonellae was 100%, even after washing. Biofilm formation occurred more on wood (60%) and plastic (40%) than glass (10%) boards, demonstrating that bacteria adhered more to a hydrophobic material. It was concluded that the cutting boards represent a critical point in cross-contamination, particularly in the presence of biofilm. Salmonella Enteritidis was able to form a biofilm on these three types of cutting boards but glass showed the least formation.

Research Salmonella Biofilm Resistance

ASMOrg Salmonella

Salmonellosis is the second most common cause of food-borne illness worldwide. Contamination of surfaces in food processing environments may result in biofilm formation with a risk of food contamination. Effective decontamination of biofilm contaminated surfaces is challenging. Using the CDC biofilm reactor, the activity of sodium hypochlorite, sodium hydroxide and benzalkonium chloride were examined against an early (48 hours) and relatively mature (168 hours) Salmonella biofilm. All 3 agents result in reduction in viable counts of Salmonella, however only sodium hydroxide resulted in eradication of the early biofilm. None of the agents achieved eradication of mature biofilm, even at 90-minutes contact time. Studies of activity of chemical disinfection against biofilm should include assessment of activity against mature biofilm. The difficulty of eradication of established Salmonella biofilm serves to emphasise the priority of preventing access of Salmonella to post-cook areas of food production facilities.