Category Archives: Biofilm

Research – Effect of food environment on the ability of microorganisms to form biofilms

Wiley Online

Abstract

The aim of this study is to understand the effect of the food product’s, even in trace amounts, in the biofilm formation potential of Escherichia coli and Staphylococcus aureus. Bacterial adhesion was tested using three different food media, in which bacterial cells were adapted before determining how this would affect their adhesion score and biofilm formation potential: sterilized milk, mineral water, and pasteurized commercial orange juice. After characterization of the bacteria and the adhesion support by the contact angle, the adhesion score obtained by the theoretical mathematical model XDLVO developed by van Oss was compared to that obtained by the experimental approach, in order to examine the role of non-specific (thermodynamic) and specific (biological) interactions in adhesion. Changes in the membrane composition of bacteria, as a function of acclimation conditions, were evaluated by infrared spectroscopy. Results reveal variability in the spectral region between 900 and 1200 cm−1 specific to capsular polysaccharides (CPS) or lipopolysaccharides (LPS), and also in the range between 2800 and 3000 cm−1 characteristic of membrane lipids and phospholipids. Biofilm formation by S. aureus and E. coli was affected by commercial UHT milk. This led to increased hydrophobicity and total adhesion energy within the environment of these strains. However, no correlation was observed between the adhesion scores obtained through experimental and theoretical approaches. This underscores the necessity of considering the specific characteristics of individual food products in understanding the phenomenon of biofilm formation. Understanding, the mechanism, the variations in bacterial surface characteristics, and the conditions that favor or disadvantage the formation of biofilms, may have important implications, in the development of preventive and curative strategies, to control of food-borne infections.

Research – Detection of Extended Spectrum ß-Lactamase-Producing Escherichia coli with Biofilm Formation from Chicken Meat in Istanbul

MDPI

Abstract

Antimicrobial resistance is one of the major public health problems worldwide. This study aimed to detect the presence of extended-spectrum β-lactamase-(ESBL-)producing Escherichia (E.coli in chicken meat in Istanbul, Türkiye. Raw chicken meat samples (n = 208) were collected from different sale points and analyzed for ESBL-producing E. coli. In total, 101 (48.5%) isolates were confirmed as E. coli by PCR, of which 80/101 (79.2%) demonstrated multiple antibiotic resistance. Resistance against amoxicillin-clavulanic acid was most frequent (87.1%). Eighteen isolates (17.8%) demonstrated phenotypical ESBL resistance, as assessed by the double disc synergy test (DDST). Isolates were tested for the presence of β-lactamase genes and mobilized colistin-resistant genes. The blaTEM group was most frequently detected (97.02%), followed by blaCTX m (45.5%), blaSHV (9.9%), and blaOXA-2 (0.9%). However, mcr genes and blaNDM, blaKPCblaVIM, and blaOXA-48 genes were not found in any isolate. E. coli strains were tested for biofilm formation in six different media [Nutrient broth, LB broth, Tryptone Soya broth (TSB), TSB containing 1% sucrose, TSB containing 0.6% yeast extract, and BHI]. Biofilm formation by E. coli isolates (44/101, 43.5%) was highest in TSB with 1% sucrose. It is worth noting that all biofilm-producing isolates were found to harbor the blaTEM-1 gene, which can indicate a high level of antibiotic resistance. This is the first report about ESBL-producing E. coli in poultry meat, the exposure of consumers in Istanbul metropolitan areas, and the ability of E. coli from this region to produce biofilms.

Research – Susceptibility of Salmonella enterica Typhimurium dry surface biofilms to disinfection

Wiley Online

Abstract

In food preparation and manufacturing environments, surfaces contaminated with Salmonella can lead to outbreaks of Salmonellosis. We hypothesise that Salmonella resides on dry surfaces in a biofilm form leading to potential environmental persistence and transfer following contact. This is the first study reporting that Salmonella Typhimurium can form dry surface biofilm (DSB). Six disinfectants commonly used in the food industry were evaluated for their efficacy against the DSB. The two most efficacious formulations reduced bacterial viability in DSB by >99.99% when combined with mechanical removal (5 sec wiping; 300 g weight). Five out of six formulations significantly reduced bacterial transfer when combined with wiping. Complete eradication of Salmonella Typhimurium DSB was challenging, and mechanical removal was essential to produce a >99.99% reduction in bacterial viability within DSB. This study highlights a potential mode of survival of Salmonella Typhimurium on food-contact surfaces and DSB challenges for disinfection.

Research – Differences in Biofilm Formation of Listeria monocytogenes and Their Effects on Virulence and Drug Resistance of Different Strains

MDPI

Abstract

Listeria monocytogenes is recognized as one of the primary pathogens responsible for foodborne illnesses. The ability of L. monocytogenes to form biofilms notably increases its resistance to antibiotics such as ampicillin and tetracycline, making it exceedingly difficult to eradicate. Residual bacteria within the processing environment can contaminate food products, thereby posing a significant risk to public health. In this study, we used crystal violet staining to assess the biofilm-forming capacity of seven L. monocytogenes strains and identified ATCC 19112 as the strain with the most potent biofilm-forming. Subsequent fluorescence microscopy observations revealed that the biofilm-forming capacity was markedly enhanced after two days of culture. Then, we investigated into the factors contributing to biofilm formation and demonstrated that strains with more robust extracellular polymer secretion and self-agglutination capabilities exhibited a more pronounced ability to form biofilms. No significant correlation was found between surface hydrophobicity and biofilm formation capability. In addition, we found that after biofilm formation, the adhesion and invasion of cells were enhanced and drug resistance increased. Therefore, we hypothesized that the formation of biofilm makes L. monocytogenes more virulent and more difficult to remove by antibiotics. Lastly, utilizing RT-PCR, we detected the expression levels of genes associated with biofilm formation, including those involved in quorum sensing (QS), flagellar synthesis, and extracellular polymer production. These genes were significantly upregulated after biofilm formation. These findings underscore the critical relationship between extracellular polymers, self-agglutination abilities, and biofilm formation. In conclusion, the establishment of biofilms not only enhances L. monocytogenes’ capacity for cell invasion and adhesion but also significantly increases its resistance to drugs, presenting a substantial threat to food safety.

Research – Inhibition and eradication of Listeria monocytogenes biofilm using the combined treatment with nisin and sesamol

Science Direct

Abstract

Listeria monocytogenes can form biofilms, which enables it to persist in the food industry and poses a significant threat to food safety. The combination of nisin (NS) and sesamol (SE) has been found to inhibit the proliferation of L. monocytogenes. The objective of this study is to evaluate the effects of the combined use of NS and SE on the biofilm formation of L. monocytogenes and investigate the potential underlying mechanisms. The results showed that the combination of NS and SE completely inhibited the biofilm formation of both strains on the stainless steel sheet within 48 h. The existing L. monocytogenes biofilm on the stainless steel sheet was completely removed within 4 h after the combined treatment of NS and SE. Compared with individual treatment, the combined use of NS and SE resulted in a more significant downregulation in gene expression of fbpA, which is associated with L. monocytogenes biofilm formation. The findings suggest that the combined strategy of NS and SE in food processing has greater potential to control the formation of L. monocytogenes biofilms and eliminate existing ones, compared with using NS or SE alone.

Research – Using Essential Oils to Reduce Yersinia enterocolitica in Minced Meat and in Biofilms

MDPI

Abstract

Yersiniosis, one of the leading foodborne infections in the European Union, is caused by Yersinia enterocolitica. In this study, the antibacterial and antibiofilm effects of cinnamon (Cinnamomum zeylanicum Nees), clove (Syzygium aromaticum L.), oregano (Origanum vulgare L.), rosemary (Rosmarinus officinalis L.), thyme (Thymus vulgaris L.), and winter savory (Satureja montana L.) essential oils were investigated against Y. enterocolitica strains belonging to the bioserotype 4/O:3. Cinnamon essential oil showed the highest antibacterial activity, with an MIC value 0.09 µL/mL, followed by oregano and thyme essential oils, with MIC values from 0.09 to 0.18 µL/mL, and from 0.18 to 0.23 µL/mL, respectively. Thyme essential oil at 0.23 µL/g (MIC) and at 0.46 µL/g (2MIC) significantly (p < 0.05) reduced the number of Y. enterocolitica by 0.38 log CFU/g and 0.64 log CFU/g, respectively, in minced pork meat during storage at 4 °C for 4 days. The Y. enterocolitica strains formed biofilms at 15 °C and 37 °C in tryptic soy broth and Luria–Bertani broth, while no biofilms were obtained at 5 °C, and in meat broth nutrient media. Applying the minimum bactericidal concentrations of cinnamon, clove, oregano, rosemary, thyme, and winter savory essential oils on preformed biofilms led to significant reductions being observed in the range from 45.34% to 78.89%. A scanning electron microscopy assay showed the devastating impact of oregano and thyme essential oils on the morphology of Y. enterocolitica bacterial cells. In conclusion, the results of this study show that essential oils possess high anti-Yersinia and antibiofilm effects.

Research – Assessing Biofilm Formation and Resistance of Vibrio parahaemolyticus on UV-Aged Microplastics in Aquatic Environments

Science Direct

Abstract

UV degradation of marine microplastics (MPs) could increase their vector potential for pathogenic bacteria and threaten human health. However, little is known about how the degree of UV aging affects interactions between MPs and pathogens and how various types of MPs differ in their impact on seafood safety. This study investigated five types of UV-aged MPs and their impact on Vibrio parahaemolyticus, a seafood pathogen. MPs exposed to UV for 60 days showed similar physicochemical changes such as surface cracking and hydrophobicity reduction. Regardless of the type, longer UV exposure of MPs resulted in more biofilm formation on the surface under the same conditions. V. parahaemolyticus types that formed biofilms on the MP surface showed 1.4- to 5.0-fold upregulation of virulence-related genes compared to those that did not form biofilms, independently of UV exposure. However, longer UV exposure increased resistance of V. parahaemolyticus on MPs to chlorine, heat, and human gastrointestinal environment. This study implies that the more UV degradation occurs on MPs, the more microbial biofilm formation is induced, which can significantly increase virulence and environmental resistance of bacteria regardless of the type of MP.

Research – Identification of Biofilm-Forming Foodborne Pathogens and Development of Prevention Strategies

Food Safety.com

A recent study emerged from Penn State University about multispecies biofilms that provide Listeria monocytogenes with a strategy against sanitizers.1 This study brings much-needed attention to a persistent problem in the food safety community. Here, I provide several comments on this topic. (see link above)

What is Biofilm? Understanding Structure and Function

Research- Effect of gas ultrafine bubbles on the potency of antimicrobials against Escherichia coli O157:H7 biofilms on various food processing surfaces

Wiley Online

Abstract

This study investigated the impact of incorporating gas [air, carbon dioxide (CO2), and nitrogen (N2)] UFB on the potency of chlorine (Cl2; 50, 100, and 200 ppm) and peracetic acid (PAA; 20, 40, and 80 ppm) antimicrobial (AM) solutions against fresh (3 days) and aged (30 days) E. coli O157:H7 biofilms on polypropylene, silicone, and stainless-steel surfaces. The biofilms were statically grown on polypropylene, silicone, and stainless-steel coupons (7.62 × 2.54 cm) at 25°C for 3 or 30 days by immersing in a 3-strain cocktail of E. coli. The incorporation of air, CO2, and N2 UFB in AM solutions resulted in significantly increased log reductions (2.1–3.7 logs) in fresh and aged E. coli biofilms on all surfaces compared to solutions without UFB, except for N2 UFB on aged stainless-steel biofilms and air UFB on aged polypropylene biofilms, which resulted in similar log reductions as solutions without UFB (1.5–2.1 logs).

Research – Effect of Bacteriophages against Biofilms of Escherichia coli on Food Processing Surfaces

MDPI

Abstract

The bacterial adhesion to food processing surfaces is a threat to human health, as these surfaces can serve as reservoirs of pathogenic bacteria. Escherichia coli is an easily biofilm-forming bacterium involved in surface contamination that can lead to the cross-contamination of food. Despite the application of disinfection protocols, contamination through food processing surfaces continues to occur. Hence, new, effective, and sustainable alternative approaches are needed. Bacteriophages (or simply phages), viruses that only infect bacteria, have proven to be effective in reducing biofilms. Here, phage phT4A was applied to prevent and reduce E. coli biofilm on plastic and stainless steel surfaces at 25 °C. The biofilm formation capacity of phage-resistant and sensitive bacteria, after treatment, was also evaluated. The inactivation effectiveness of phage phT4A was surface-dependent, showing higher inactivation on plastic surfaces. Maximum reductions in E. coli biofilm of 5.5 and 4.0 log colony-forming units (CFU)/cm2 after 6 h of incubation on plastic and stainless steel, respectively, were observed. In the prevention assays, phage prevented biofilm formation in 3.2 log CFU/cm2 after 12 h. Although the emergence of phage-resistant bacteria has been observed during phage treatment, phage-resistant bacteria had a lower biofilm formation capacity compared to phage-sensitive bacteria. Overall, the results suggest that phages may have applicability as surface disinfectants against pathogenic bacteria, but further studies are needed to validate these findings using phT4A under different environmental conditions and on different materials.