Comparative Assessment of the Antibacterial and Antibiofilm Actions of Benzalkonium Chloride, Erythromycin, and L(+)-Lactic Acid against Raw Chicken Meat Campylobacter spp. Isolates.

Campylobacter spp. are significant zoonotic agents, which cause annually millions of human cases of foodborne gastroenteritis worldwide. Their inclusion in biofilms on abiotic surfaces seems to play a pivotal role in their survival outside of the host, growth, and spread. To successfully mitigate the risks that arise with these bacteria, it is crucial to decrease their prevalence within the food production chain (from farm to the table), alongside the successful treatment of the resulting illness, known as campylobacteriosis. For this, the use of various antimicrobial agents remains actively in the foreground. A general-purpose biocide and cationic surfactant (benzalkonium chloride; BAC), a widely used macrolide antibiotic (erythromycin; ERY), and a naturally occurring organic acid (L(+)-lactic acid; LA) were comparatively evaluated in this work for their potential to inhibit both the planktonic and biofilm growth of 12 selected Campylobacter spp. (of which, seven were C. jejuni and five were C. coli) raw chicken meat isolates, all grown in vitro as monocultures. The inhibitory action of LA was also studied against four mixed-culture Campylobacter biofilms (each composed of three different isolates). The results showed that the individual effectiveness of the agents varied significantly depending on the isolate, growth mode (planktonic, biofilm), intercellular interactions (monocultures, mixed cultures), and the growth medium used (with special focus on blood presence). Thus, BAC exhibited minimum inhibitory concentrations (MICs), minimum bactericidal concentrations (MBCs), and minimum biofilm inhibitory concentrations (MBICs) that ranged from 0.5 to 16 µg/mL. Interestingly enough, these values varied widely from 0.25 to 1024 µg/mL for ERY. Concerning LA, the MICs, MBCs, and MBICs varied from 1024 to 4096 µg/mL, with mixed-culture biofilm formation always being more difficult to suppress when compared to biofilm monocultures. In addition, it was evident that intercellular interactions encountered within mixed-culture Campylobacter biofilms significantly influenced both the population dynamics and the tolerance of each consortium member to acid exposure. Overall, the findings of this study provide useful information on the comparative effectiveness of three well-known antimicrobial agents for the control of Campylobacter spp. under various growth modes (i.e., planktonic, biofilm, monocultures, mixed cultures) that could potentially be encountered in food production and clinical settings.

Development of a functional Greek sheep yogurt incorporating a probiotic Lacticaseibacillus rhamnosus wild-type strain as adjunct starter culture.

Greek yogurt is a fermented dairy product of high nutritional value that can be used as a matrix for the delivery of probiotics. The aim of this study was to develop a new probiotic Greek sheep yogurt with upgraded quality and functional characteristics. To do this, yogurt was manufactured by fermenting pasteurized milk with the commercial starter culture (Streptococcus thermophilus (ST), Lactobacillus bulgaricus (LB)) together with a probiotic Lacticaseibacillus rhamnosus (LR) wild-type strain (probiotic yogurt; PY). As a control, yogurt manufactured with only the starter culture (ST, LB) was used (conventional yogurt; CY) The survival of all three lactic acid bacteria (LAB) species (ST, LB, and LR) was monitored throughout the products' shelf life (storage at 4 °C for 25 days), and also following exposure to a static in vitro digestion model (SIVDM). The population dynamics of total aerobic plate count (APC), Enterobacteriaceae, yeasts and molds grown in both yogurts were also determined. The total antioxidant activity (AA) of yogurts was comparatively determined using in parallel two different assays, whereas the Folin-Ciocalteu assay was used to determine their total phenolic content (TPC). At each sampling day, yogurts were also evaluated for their pH, titratable acidity (TA) and main sensory characteristics. The population of probiotic LR remained stable during the shelf life (and above 108 CFU/g). Yogurt starters (ST, LB) were not detected following SIVDM, whereas LR (in PY) presented a reduction of about only one log. The AA and TPC of PY were found significantly higher than that of CY (P < 0.05). At the end of storage (25th day), neither pH nor TA differed significantly between the two yogurt types, while no fungal growth was observed in the PY. Consumer sensory analysis did not reveal important differences between the two yogurt types during their shelf life. To sum up, the novel yogurt was able to deliver to consumers a high number of probiotic cells (>108 CFU/g), presented increased antioxidant power, had an expanded shelf life, and maintained its good sensory attributes.

Prevalence of Campylobacter spp., Salmonella spp., and Listeria monocytogenes, and Population Levels of Food Safety Indicator Microorganisms in Retail Raw Chicken Meat and Ready-To-Eat Fresh Leafy Greens Salads Sold in Greece.

The presence of microbial pathogens in foods compromises their safety resulting in foodborne illnesses, public health disorders, product recalls, and economic losses. In this work, 60 samples of chilled raw chicken meat and 40 samples of packaged ready-to-eat (RTE) fresh leafy greens salads, sold in Greek retail stores (butchers and supermarkets), were analyzed for the presence of three important foodborne pathogenic bacteria, i.e., Campylobacter spp., Salmonella spp., and Listeria monocytogenes, following the detection protocols of the International Organization for Standardization (ISO). In parallel, the total aerobic plate count (APC), Enterobacteriaceae, total coliforms, Escherichia coli, and staphylococci were also enumerated as hygiene (safety) indicator organisms. When present, representative typical colonies for each pathogen were biochemically verified, following the ISO guidelines. At the same time, all the Campylobacter isolates from chicken (n = 120) were identified to the species level and further phylogenetically discriminated through multiplex and repetitive sequence-based (rep) polymerase chain reaction (PCR) methods, respectively. Concerning raw chicken, Campylobacter spp. were recovered from 54 samples (90.0%) and Salmonella spp. were recovered from 9 samples (15.0%), while L. monocytogenes was present in 35 samples (58.3%). No Campylobacter was recovered from salads, and Salmonella was present in only one sample (2.5%), while three salads were found to be contaminated with L. monocytogenes (7.5%). The 65% of the Campylobacter chicken isolates belonged to C. jejuni, whereas the rest, 35%, belonged to C. coli. Alarmingly, APC was equal to or above 106 CFU/g in 53.3% and 95.0% of chicken and salad samples, respectively, while the populations of some of the other safety indicators were in some cases also high. In sum, this study unravels high occurrence percentages for some pathogenic and food safety indicator microorganisms in raw chicken meat and RTE fresh leafy greens salads sold in Greek retail, highlighting the need for more extensive microbiological control throughout the food production chain (from the farm/field to the market).

Kombucha Fermentation of Olympus Mountain Tea (Sideritis scardica) Sweetened with Thyme Honey: Physicochemical Analysis and Evaluation of Functional Properties.

This study implemented kombucha fermentation of Olympus Mountain tea (Sideritis scardica) sweetened with honey (OMTWH) in order to investigate the potential for producing a novel beverage with functional properties. The increase in the total count of bacteria and yeast suggests that the OMTWH acts as a viable substrate for supporting the proliferation of the microorganisms of the Kombucha symbiotic culture. The fermentation resulted in a reduction in pH and increased total titratable acidity. After fermentation, a statistically significant increase in the vitamins C, B1, B2, B6, B7, and B12 content was observed (p < 0.05). Total phenolics and antioxidant activity of the fermented beverage was significantly enhanced, as assessed by the method of Folin-Ciocalteu and ABTS assay, respectively. Results revealed that OMTWH had a potent inhibitory activity of α-amylase, α-glucosidase, acetylcholinesterase, and butyrylcholinesterase; OMTWH fermented with a kombucha consortium exhibited even higher inhibition. Hence, the process of kombucha fermentation can convert OMTWH into a novel beverage with enhanced functional properties.

Chemical Composition, Antibacterial and Antibiofilm Actions of Oregano (Origanum vulgare subsp. hirtum) Essential Oil against Salmonella Typhimurium and Listeria monocytogenes.

Essential oils (EOs) are plant mixtures that are known to present strong bioactivities, including a wide antimicrobial action. Biofilms are microbial sessile structures that represent the default mode of growth of microorganisms in most environments. This study focused on the antimicrobial action of the EO extracted from one of the most representative oregano species, that is, Origanum vulgare (subsp. hirtum), against two important foodborne pathogens, Salmonella enterica (serovar Typhimurium) and Listeria monocytogenes. For this, the minimum inhibitory concentrations of the EO against the planktonic and biofilm growth of each bacterium were determined (MICs, MBICs), together with the minimum bactericidal and biofilm eradication concentrations (MBCs, MBECs). The EO was also analyzed for its chemical composition by gas chromatography-mass spectrometry analysis (GC-MS). The influence of EO exposure on the expression of some important virulence genes (hly, inlA, inlB and prfA) was also studied in L. monocytogenes. Results revealed a strong antibacterial and antibiofilm action with MICs and MBICs ranging from 0.03% to 0.06% (v/v) and from 0.06% to 0.13% (v/v), respectively. The application of the EO at 6.25% (v/v) for 15 min resulted in the total eradication of the biofilm cells of both pathogens. The EO was mainly composed of thymol, p-cymene, γ-terpinene and carvacrol. The 3 h exposure of L. monocytogenes planktonic cells to the EO at its MBIC (0.06% v/v) resulted in the significant downregulation of all the studied genes (p < 0.05). To sum, the results obtained advocate for the further exploitation of the antimicrobial action of oregano EO in food and health applications.

Chemical Composition, Antioxidant, and Antibiofilm Properties of Essential Oil from Thymus capitatus Plants Organically Cultured on the Greek Island of Lemnos.

Essential oils (EOs) are mixtures of volatile plant secondary metabolites and have been exploited by humans for thousands of years for various purposes because of their many bioactivities. In this study, the EO from Thymus capitatus, a thyme species organically cultured on the Greek Island of Lemnos, was analyzed for its chemical composition (through GC-FID and GC-MS), antioxidant activity (AA), and total phenolic content (TPC), as well as its antimicrobial and antibiofilm actions against three important foodborne bacterial pathogens (Salmonella enterica ser. Typhimurium, Listeria monocytogenes, and Yersinia enterocolitica). For the latter investigations, the minimum inhibitory concentrations (MICs) and minimum biofilm inhibitory concentrations (MBICs) of the EO against the planktonic and biofilm growth of each pathogen were determined, together with the minimum biofilm eradication concentrations (MBECs). Results revealed that T. capitatus EO was rich in thymol, p-cymene, and carvacrol, presenting high AA and TPC (144.66 µmol TroloxTM equivalents and 231.32 mg gallic acid equivalents per g of EO, respectively), while its MICs and MBICs ranged from 0.03% to 0.06% v/v and 0.03% to 0.13% v/v, respectively, depending on the target pathogen. The EO was able to fully destroy preformed (mature) biofilms of all three pathogenic species upon application for 15 min, with MBECs ranging from 2.00 to 6.25% v/v. Overall, the results demonstrate that the EO of organically cultured T. capitatus presents strong antioxidant, antibacterial, and antibiofilm properties and could, therefore, be further exploited as a functional and antimicrobial natural formulation for food and health applications.

Marked inter-strain heterogeneity in the differential expression of some key stress response and virulence-related genes between planktonic and biofilm cells in Listeria monocytogenes.

Listeria monocytogenes is a facultatively intracellular pathogenic bacterium that can provoke invasive listeriosis, a severe foodborne infection in humans. Outside the host, this is capable to survive for long periods in soil, and water, as well as on plants, while, like many other microorganisms, this can also attach to abiotic surfaces, such as food contact ones, forming biofilms on them. It has been suggested that inside those sessile communities, L. monocytogenes cells not only display an increased stress tolerance but may also boost their pathogenicity. In this work, the expression of ten key stress response and/or virulence-related genes (i.e., groEL, hly, iap, inlA, inlB, lisK, mdrD, mdrL, prfA, and sigB) was studied in three different L. monocytogenes strains (AAL20066, AAL20107, and PL24), all isolated from foods and each belonging to a different listeriosis-associated serovar (1/2a, 1/2b, and 1/2c, respectively). For this, each strain was initially left to develop a mature biofilm on a model polystyrene surface (Petri dish) by incubating for 144 h (6 days) at 20 °C in tryptone soya broth (with medium renewal every 48 h). Following incubation, both biofilm and the surrounding free-swimming (planktonic) cells were recovered, and their gene expressions were comparatively evaluated through targeted reverse transcription-quantitative polymerase chain reactions (RT-qPCR). Results revealed a strain-dependent differential gene expression between the two cell types. Thus, for instance, in strain AAL20107 (ser. 1/2b) biofilm growth worryingly resulted in a significant overexpression of all the studied genes (P < 0.05), whereas in strain PL24 (ser. 1/2c), the expression of most genes (8/10) did not change upon biofilm growth, with only two of them (groEL and hly) being again significantly upregulated. Such transcriptomic strain variability in stress adaptation and/or virulence induction should be generally considered in the physiological studies of pathogenic biofilms and preferably upon designing and implementing novel and more efficient eradication methods.

The transmittable through stinging microbiota differs between honeybees and wasps: a potentially greater microbial risk of the wasp sting for humans.

The present research investigated whether accidental contact through stinging with honeybees, wasps, and hornets could represent a microbial hazard for humans. It has been previously suggested that such contact may transmit pathogens causing infections that could even be fatal for some susceptible individuals. Stinging simulation experiments were performed in the lab with live insects collected from the environment in Lemnos Island (north-eastern Greece), while different selective agar media targeting some clinically important bacteria (i.e., Staphylococcus aureus, Streptococcus pyogenes, Enterococcus faecalis/faecium, and Pseudomonas aeruginosa) were used as substrates for microbial recovery and identification. Results revealed none of the target pathogenic bacterial species in the honeybee samples, with bacilli, staphylococci, and micrococci dominating their surveyed microbiota. However, most of the suspect colonies isolated from wasps and hornets belonged to important hygienic indicators (i.e., enterococci, Proteus mirabilis, and coliforms), implying possible contact of these insects with fecal origin materials. To sum up, the microbiota that may be transmitted to humans through stinging appears to differ between honeybees and wasps/hornets, while the isolation from the latter samples of some other important opportunistic pathogens, such as Enterobacter spp. and Klebsiella spp., also known for multidrug resistance, could be an additional reason of concern.

Relevance and Importance of Biofilms in the Resistance and Spreading of Campylobacter spp. Within the Food Chain.

Campylobacter spp. are Gram-negative microaerophilic and non-spore-forming bacteria that primarily live as commensal organisms in the gastrointestinal tract of many domestic and wild birds and mammals. These frequently provoke foodborne illness, being responsible for the most common cause of acute bacterial gastroenteritis worldwide. Although these are typically fastidious and slow-growing microorganisms which are sensitive to desiccation and other stresses (e.g., extreme pH, freezing, UV, disinfectants), these can still survive outside their host for long periods, with their attachment to surfaces and the formation of and/or inclusion into biofilms to be considered quite important for their environmental survival and widespread dissemination. In this chapter, the most representative studies on the existence, relevance, and importance of biofilms in the resistance and spreading of Campylobacter spp. within the food chain are presented. Hopefully, such accumulated and focused knowledge is expected to highlight the problem and trigger more effective ways for its mitigation, improving the safety of our food supply and protecting public health.

Investigating Possible Synergism in the Antioxidant and Antibacterial Actions of Honey and Propolis from the Greek Island of Samothrace through Their Combined Application.

Several honeybee products are known for their functional properties, including important antioxidant and antimicrobial actions. The present study examines the antioxidant activity (AA), total polyphenolic content (TPC), and antibacterial action of honey and propolis samples collected from the Greek island of Samothrace, which were applied in vitro either individually or in combination in selected concentrations. To accomplish this, the 2,2-diphenyl-1-picrylhydrazyl (DPPH) scavenging activity and the Folin-Ciocalteu assays were employed to determine the AA and TPC, respectively, while the antibacterial action was investigated against each one of four important pathogenic bacterial species causing foodborne diseases (i.e., Salmonella enterica, Yersinia enterocolitica, Staphylococcus aureus, and Listeria monocytogenes) using the agar well diffusion assay. Compared to honey, propolis presented significantly higher AA and TPC, while its combined application with honey (at ratios of 1:1, 3:1, and 1:3) did not increase these values. Concerning the antibacterial action, Y. enterocolitica was proven to be the most resistant of all the tested bacteria, with none of the samples being able to inhibit its growth. S. enterica was susceptible only to the honey samples, whereas L. monocytogenes only to the propolis samples. The growth of S. aureus was inhibited by both honey and propolis, with honey samples presenting significantly higher efficacy than those of propolis. Νo synergism in the antibacterial actions was observed against any of the tested pathogens. Results obtained increase our knowledge of some of the medicinal properties of honey and propolis and may contribute to their further exploitation for health promotion and/or food-related applications (e.g., as preservatives to delay the growth of pathogenic bacteria).

Applications and Prospects of Nanotechnology in Food and Cosmetics Preservation.

Cosmetic and food products containing water are prone to contamination during the production, storage, and transit process, leading to product spoilage and degraded organoleptic characteristics. The efficient preservation of food and cosmetics is one of the most important issues the industry is facing today. The use of nanotechnology in food and cosmetics for preservation purposes offers the possibility to boost the activity of antimicrobial agents and/or promote their safer distribution into the end product upon incorporation into packaging or film constructions. In this review, current preservation strategies are discussed and the most recent studies in nanostructures used for preservation purposes are categorized and analyzed in a way that hopefully provides the most promising strategies for both the improvement of product safety and shelf-life extension. Packaging materials are also included since the container plays a major role in the preservation of such products. It is conclusively revealed that most of the applications refer to the nanocomposites as part of the packaging, mainly due to the various possibilities that nanoscience offers to this field. Apart from that, the route of exposure being either skin or the gastrointestinal system involves safety concerns, and since migration of nanoparticles (NPs) from their container can be measured, concerns can be minimized. Conclusion: Nanomaterial science has already made a significant contribution to food and cosmetics preservation, and rapid developments in the last years reinforce the belief that in the future much of the preservation strategies to be pursued by the two industries will be based on NPs and their nanocomposites.

Εvaluation of the microbial stability and shelf life of 50% NaCl-reduced traditional Greek pork meat product "Syglino of Monemvasia" stored under vacuum at different temperatures.

Nowadays, consumers are increasingly concerned about nutrition and health issues. "Syglino of Monemvasia" is a traditional Greek cooked and smoked, sliced pork meat product. Although this is a nutritious food, its consumption should be done in moderation due to the pickling process of its preparation. This product was thus here optimized to contain half salt (NaCl) amount and its microbial stability and shelf life was then assessed in comparison to the already available commercial product. For this, the total viable counts (TVCs) and some critical specific spoilage associations were enumerated at each product type during vacuum incubation at four different temperatures (0, 5, 10, and 15 °C). The alterations in pH, aw, color, and some other crucial sensory attributes of each product were also periodically monitored. The new low-salt product was found to remain microbiologically stable under refrigerated vacuum storage for approximately two weeks, being finally spoiled by Brochothrix thermosphacta grown above 107 CFU/g, ultimately resulting in the deterioration of taste, odor, and overall appearance of the product, and thus leading to its subsequent organoleptic rejection. Despite its limited shelf life, the 50% NaCl-reduced "Syglino" could be released in the local market provided that the cooling chain is maintained throughout its distribution (≤5 °C) and at the same time consumers are willing to accept its milder taste. Although salt replacers could have been used to improve its flavor and at the same time increase its shelf life, the product here developed without the use of such alternatives will hopefully contribute to the taste training of its consumers for less salt in their diet, keeping in parallel its clean label with no added preservatives and other food additives.

Investigating Transcriptomic Induction of Resistance and/or Virulence in Listeria monocytogenes Cells Surviving Sublethal Antimicrobial Exposure.

The potential transcriptomic induction of resistance and/or virulence in two L. monocytogenes strains belonging to the most frequent listeriosis-associated serovars (i.e., 1/2a and 4b), following their sublethal antimicrobial exposure, was studied through qPCR determination of the relative expression of 10 selected related genes (i.e., groEL, hly, iap, inlA, inlB, lisK, mdrD, mdrL, prfA, and sigB). To induce sublethal stress, three common antimicrobials (i.e., benzalkonium chloride, thymol, and ampicillin) were individually applied for 2 h at 37 °C against stationary phase cells of each strain, each at a sublethal concentration. In general, the expression of most of the studied genes remained either stable or was significantly downregulated following the antimicrobial exposure, with some strain-specific differences to be yet recorded. Thymol provoked downregulation of most of the studied genes, significantly limiting the expression of 6/10 and 4/10 genes in the strains of ser. 1/2a and ser. 4b, respectively, including those coding for the master regulators of stress response and virulence (SigB and PrfA, respectively), in both strains. At the same time, the two genes coding for the invasion internalin proteins (InlA and InlB), with crucial role in the onset of L. monocytogenes pathogenesis, were both importantly upregulated in ser. 4b strain. The results obtained increase our knowledge of the stress physiology of L. monocytogenes under certain sublethal antimicrobial conditions that could be encountered within the food chain and in clinical settings, and may assist in better and more effective mitigation strategies.

Comparing the Antimicrobial Actions of Greek Honeys from the Island of Lemnos and Manuka Honey from New Zealand against Clinically Important Bacteria.

Honey is a natural food with a long history as a traditional medicine because of its many biological characteristics, including antimicrobial, antioxidant, anti-tumor and anti-inflammatory properties. In this study, the antimicrobial actions of eight different honeys from Lemnos island (north-eastern Greece) plus manuka honey (from New Zealand, UMF 30+, licensed in many countries as topical medical preparation) were evaluated against 10 clinically relevant bacteria, including five Gram-positive and five Gram-negative. To achieve this, an agar well diffusion assay measured the diameter of inhibition zones (mm) of two selected concentrations for each honey (25% and 12.5% v/v). The minimum inhibitory and bactericidal concentrations (MIC and MBC) of each sample were also calculated and compared against two representative bacterial species (Salmonella Typhimurium and Staphylococcus aureus) using broth microdilution and agar spot methods, respectively. The pH, water activity (aw), 5-hydroxymethylfurfural (HMF) and diastase levels, together with the pollen type and content of each honey, were also determined. Results revealed that all the Lemnos honeys presented antibacterial action, which for some samples was like that of manuka. These all had an acidic pH (3.61 ± 0.04), with a aw ≤ 0.60, while it is worth noting that those found to display the strongest antibacterial actions also presented the lowest HMF content, together with the highest diastase values, both of the latter being used as quality parameters. Pollen composition of the Lemnos honeys was multifloral, underlining the rich plant biodiversity encountered on the island. To summarize, Lemnos honeys could be further exploited as natural antimicrobial systems for use in foods and medicine.

Development of a herbal mouthwash containing a mixture of essential oils and plant extracts and in vitro testing of its antimicrobial efficiency against the planktonic and biofilm-enclosed cariogenic bacterium Streptococcus mutans.

A herbal mouthwash containing essential oils of holy basil and mountain tea, extracts of St John's wort and European goldenrod (Bucovia™) and cetylpyridinium chloride, was developed and in vitro tested for its efficiency against biofilm formation by Streptococcus mutans, together with its eradicating activity against already preformed (48 h with saccharose) streptococcal biofilm. The minimum inhibitory (MIC) and bactericidal concentrations (MBC) of the final formulation, as well as of its individual components, were initially determined. The results revealed that the mouthwash needed to be applied at two-times its MIC (0.63% v.v-1) to completely inhibit biofilm formation by S. mutans, which was otherwise capable of developing a robust biofilm on the tested surface. Once fully developed, the matrix of the biofilm was found to contain a significant amount of exopolysaccharides protecting the cells, being impossible to eradicate even when exposed to pure mouthwash for 15 min, highlighting the great recalcitrance of biofilm-embedded S. mutans.

Advanced Killing Potential of Thymol against a Time and Temperature Optimized Attached Listeria monocytogenes Population in Lettuce Broth.

Fresh vegetables and salads are increasingly implicated in outbreaks of foodborne infections, such as those caused by Listeria monocytogenes, a dangerous pathogen that can attach to the surfaces of the equipment creating robust biofilms withstanding the killing action of disinfectants. In this study, the antimicrobial efficiency of a natural plant terpenoid (thymol) was evaluated against a sessile population of a multi-strain L. monocytogenes cocktail developed on stainless steel surfaces incubated in lettuce broth, under optimized time and temperature conditions (54 h at 30.6 °C) as those were determined following response surface modeling, and in comparison, to that of an industrial disinfectant (benzalkonium chloride). Prior to disinfection, the minimum bactericidal concentrations (MBCs) of each compound were determined against the planktonic cells of each strain. The results revealed the advanced killing potential of thymol, with a concentration of 625 ppm (= 4 × MBC) leading to almost undetectable viable bacteria (more than 4 logs reduction following a 15-min exposure). For the same degree of killing, benzalkonium chloride needed to be used at a concentration of at least 20 times more than its MBC (70 ppm). Discriminative repetitive sequence-based polymerase chain reaction (rep-PCR) also highlighted the strain variability in both biofilm formation and resistance. In sum, thymol was found to present an effective anti-listeria action under environmental conditions mimicking those encountered in the salad industry and deserves to be further explored to improve the safety of fresh produce.

Bacterial coaggregation in aquatic systems.

The establishment of a sessile community is believed to occur in a sequence of steps where genetically distinct bacteria can become attached to partner cells via specific molecules, in a process known as coaggregation. The presence of bacteria with the ability to autoaggregate and coaggregate has been described for diverse aquatic systems, particularly freshwater, drinking water, wastewater, and marine water. In these aquatic systems, coaggregation already demonstrated a role in the development of complex multispecies sessile communities, including biofilms. While specific molecular aspects on coaggregation in aquatic systems remain to be understood, clear evidence exist on the impact of this mechanism in multispecies biofilm resilience and homeostasis. The identification of bridging bacteria among coaggregating consortia has potential to improve the performance of wastewater treatment plants and/or to contribute for the development of strategies to control undesirable biofilms. This study provides a comprehensive analysis on the occurrence and role of bacterial coaggregation in diverse aquatic systems. The potential of this mechanism in water-related biotechnology is further described, with particular emphasis on the role of bridging bacteria.

Common Plant-Derived Terpenoids Present Increased Anti-Biofilm Potential against Staphylococcus Bacteria Compared to a Quaternary Ammonium Biocide.

The antimicrobial actions of three common plant-derived terpenoids (i.e., carvacrol, thymol and eugenol) were compared to those of a typical quaternary ammonium biocide (i.e., benzalkonium chloride; BAC), against both planktonic and biofilm cells of two widespread Staphylococcus species (i.e., S. aureus and S. epidermidis). The minimum inhibitory and bactericidal concentrations (MICs, MBCs) of each compound against the planktonic cells of each species were initially determined, together with their minimum biofilm eradication concentrations (MBECs). Various concentrations of each compound were subsequently applied, for 6 min, against each type of cell, and survivors were enumerated by agar plating to calculate log reductions and determine the resistance coefficients (Rc) for each compound, as anti-biofilm effectiveness indicators. Sessile communities were always more resistant than planktonic ones, depending on the biocide and species. Although lower BAC concentrations were always needed to kill a specified population of either cell type compared to the terpenoids, for the latter, the required increases in their concentrations, to be equally effective against the biofilm cells with respect to the planktonic ones, were not as intense as those observed in the case of BAC, presenting thus significantly lower Rc. This indicates their significant anti-biofilm potential and advocate for their further promising use as anti-biofilm agents.