Acetic and citric acids effect the type II secretion system and decrease the metabolic activities of salmon spoilage-related Rahnella aquatilis KM05.

Rahnella aquatilis causes seafoods to spoil by metabolizing sulfur-containing amino acids and/or proteins, producing H2S in products. The type II secretion system (T2SS) regulates the transport of proteases from the cytoplasm to the surrounding environment and promotes bacterial growth at low temperatures. To prevent premature fish spoilage, new solutions for inhibiting the T2SS of bacteria should be researched. In this study, global transcriptome sequencing was used to analyze the spoilage properties of R. aquatilis KM05. Two of the mapped genes/coding sequences (CDSs) were matched to the T2SS, namely, qspF and gspE, and four of the genes/CDSs, namely, ftsH, rseP, ptrA and pepN, were matched to metalloproteases or peptidases in R. aquatilis KM05. Subinhibitory concentrations of citric (18 µM) and acetic (41 µM) acids caused downregulation of T2SS-related genes (range from - 1.0 to -4.5) and genes involved in the proteolytic activities of bacteria (range from - 0.5 to -4.0). The proteolytic activities of R. aquatilis KM05 in vitro were reduced by an average of 40%. The in situ experiments showed the antimicrobial properties of citric and acetic acids against R. aquatilis KM05; the addition of an acidulant to salmon fillets limited microbial growth. Citric and acetic acids extend the shelf life of fish-based products and prevent food waste.

Global transcriptome analysis of Pseudomonas aeruginosa NT06 response to potassium chloride, sodium lactate, sodium citrate, and microaerophilic conditions in a fish ecosystem.

Pseudomonas aeruginosa is an opportunistic pathogen that recently has been increasingly isolated from foods, especially from minimally processed fish-based products. Those are preserved by the addition of sodium chloride (NaCl) and packaging in a modified atmosphere. However, the current trends of minimizing NaCl content may result in an increased occurrence of P. aeruginosa. NaCl can be replaced with potassium chloride (KCl) or sodium salts of organic acids. Herein, we examined the antimicrobial effects of KCl, sodium lactate (NaL), sodium citrate (NaC), and sodium acetate (NaA) against P. aeruginosa NT06 isolated from fish. Transcriptome response of cells grown in medium imitating a fish product supplemented with KCl and KCl/NaL/NaC and maintained under microaerophilic conditions was analysed. Flow cytometry analysis showed that treatment with KCl and KCl/NaL/NaC resulted in changed metabolic activity of cells. In response to KCl and KCl/NaL/NaC treatment, genes related to cell maintenance, stress response, quorum sensing, virulence, efflux pump, and metabolism were differentially expressed. Collectively, our results provide an improved understanding of the response of P. aeruginosa to NaCl alternative compounds that can be implemented in fish-based products and encourage further exploration of the development of effective methods to protect foods against the P. aeruginosa, underestimate foodborne bacteria.

Potassium Chloride, Sodium Lactate and Sodium Citrate Impaired the Antimicrobial Resistance and Virulence of Pseudomonas aeruginosa NT06 Isolated from Fish.

Sodium chloride (NaCl) is a commonly used additive in minimally processed fish-based products. The addition of NaCl to fish products and packaging in a modified atmosphere is usually efficient with regard to limiting the occurrence of the aquatic environmental pathogen Pseudomonas aeruginosa. Given the negative effects of excess NaCl in the diet, there is a growing demand to reduce NaCl in food products with safer substituents, but the knowledge of their impact on antibiotic resistant P. aeruginosa is limited. This study aimed to evaluate the physiological and transcriptome characteristics of P. aeruginosa NT06 isolated from fish and to determine the effect of selected concentrations of alternative NaCl compounds (KCl/NaL/NaC) on the P. aeruginosa NT06 virulence phenotype and genotype. In the study, among the isolated microorganisms, P. aeruginosa NT06 showed the highest antibiotic resistance (to ampicillin, ceftriaxone, nalidixic acid, and norfloxacin) and the ability to grow at 4 °C. The Comprehensive Antibiotic Resistance Database (CARD) and the Virulence Factor Database (VFDB) revealed the presence of 24 and 134 gene products assigned to AMR and VF in the P. aeruginosa NT06 transcriptome, respectively. KCl, KCl/NaL and KCl/NaL/NaC inhibited pyocyanin biosynthesis, elastase activity, and protease activity from 40 to 77%. The above virulence phenotypic observations were confirmed via RT-qPCR analyses, which showed that all tested AMR and VF genes were the most downregulated due to KCl/NaL/NaC treatment. In conclusion, this study provides insight into the potential AMR and VF among foodborne P. aeruginosa and the possible impairment of those features by KCl, NaL, and NaC, which exert synergistic effects and can be used in minimally processed fish-based products.

Chlorogenic Acid Inhibits Rahnella aquatilis KM25 Growth and Proteolytic Activity in Fish-Based Products.

This work verified the antiproliferative and antiproteolytic activities of chlorogenic acid against Rahnella aquatilis KM25, a spoilage organism of raw salmon stored at 4 °C. Chlorogenic acid limited the growth of R. aqatilis KM25 in vitro at a concentration of 2.0 mg/mL. The dead (46%), viable (25%), and injured (20%) cell subpopulations were identified by flow cytometry following treatment of R. aquatilis KM25 with the examined agent. The exposure of R. aquatilis KM25 to chlorogenic acid altered its morphology. Changes in cell dimensions, mostly in length parameters from 0.778 µm to 1.09 µm, were found. The length of untreated cells ranged from 0.958 µm to 1.53 µm. The RT-qPCR experiments revealed changes in the expression of genes responsible for the proliferation and proteolytic activity of cells. Chlorogenic acid caused a significant reduction in the mRNA levels of the ftsZ, ftsA, ftsN, tolB, and M4 genes (-2.5, -1.5, -2.0, -1.5, and -1.5, respectively). In situ experiments confirmed the potential of chlorogenic acid to limit bacterial growth. A similar effect was noted in samples treated with benzoic acid, where the growth inhibition of R. aquatilis KM25 was 85-95%. Reduction of microbial R. aquatilis KM25 proliferation significantly limited total volatile base nitrogen (TVB-N) and trimethylamine (TMA-N) formation during storage, extending the shelf life of model products. The TVB-N and TMA-N parameters did not exceed the upper levels of the maximum permissible limit of acceptability. In this work, the TVB-N and TMA-N parameters were 10-25 mg/100 g and 2.5-20.5 mg/100 g, respectively; for samples with benzoic acid-supplemented marinades, the parameters TVB-N and TMA-N were 7.5-25.0 mg/100 g and 2.0-20.0 mg/100 g, respectively. Based on the results of this work, it can be concluded that chlorogenic acid can increase the safety, shelf life, and quality of fishery products.

The Combined Effect of Lactic Acid Bacteria and Galactomyces geotrichum Fermentation on the Aroma Composition of Sour Whey.

The increase in demand for food flavorings due to the shortening and simplification of food production technology also entails an increase in the demand for new technologies for their production. The biotechnological production of aromas is a solution characterized by a high efficiency, an independence from environmental factors and a relatively low cost. In this study, the influence of the implementation of lactic acid bacteria pre-fermentation into the production of aroma compounds by Galactomyces geotrichum on a sour whey medium on the intensity of the obtained aroma composition was analyzed. The monitoring of the culture in terms of biomass buildup, the concentration of selected compounds, and the pH resulted in the confirmation of interactions between the analyzed microorganisms. The post-fermentation product underwent a comprehensive sensomic analysis for the identification and quantification of the aroma-active compounds. The use of gas chromatography-olfactometry (GC-O) analysis and the calculation of odor activity values (OAVs) allowed 12 key odorants to be identified in the post-fermentation product. The highest OAV was found for phenylacetaldehyde with a honey odor (1815). The following compounds with the highest OAVs were 2,3-butanedione with a buttery aroma (233), phenylacetic acid with a honey aroma (197), 2,3-butanediol with a buttery aroma (103), 2-phenylethanol with a rosy aroma (39), ethyl octanoate with a fruity aroma (15), and ethyl hexanoate with a fruity aroma (14).

Gallic and ferulic acids suppress proteolytic activities and volatile trimethylamine production in the food-borne spoiler Rahnella aquatilis KM05.

BACKGROUND: Rahnella aquatilis is a recognised microbial threat that alters the sensory properties of seafood. The high frequency with which R. aquatilis is isolated from fish has prompted a search for alternative preservatives. In the present study, in vitro and fish-based ecosystem (raw salmon-based medium) approaches were used to validate the antimicrobial effects of gallic (GA) and ferulic (FA) acids against R. aquatilis KM05. The results were compared with data describing the response of KM05 to sodium benzoate. Bioinformatics data of the whole genome were used to analyse the potential for fish spoilage by KM05 in detail, and the results revealed the main physiological characteristics that underlie reduced seafood quality. RESULTS: In the KM05 genome, the most abundantly enriched Gene Ontology terms were 'metabolic process', 'organic substance metabolic process' and 'cellular process'. Through an evaluation of the Pfam annotations, 15 annotations were found to be directly involved in the proteolytic activity of KM05. Peptidase_M20 was the most abundantly represented (abundance value of 14060). Proteins representing the CutC family (abundance value of 427) indicated the potential for KM05 degradation of trimethyl-amine-N-oxide. Subinhibitory concentrations of GA and FA suppressed the proteolytic activities of KM05 both in vitro and in RS medium by an average of 33-45%. These results were confirmed by quantitative real-time PCR experiments, which also showed that the expression levels of genes involved in proteolytic activities and volatile trimethylamine production were also decreased. CONCLUSION: Phenolic compounds can be used as potential food additives for preventing quality deterioration of fish products. © 2023 Society of Chemical Industry.

Black pepper and tarragon essential oils suppress the lipolytic potential and the type II secretion system of P. psychrophila KM02.

Given the increasing consumer demand for raw, nonprocessed, safe, and long shelf-life fish and seafood products, research concerning the application of natural antimicrobials as alternatives to preservatives is of great interest. The aim of the following paper was to evaluate the effect of essential oils (EOs) from black pepper (BPEO) and tarragon (TEO), and their bioactive compounds: limonene (LIM), ß-caryophyllene (CAR), methyl eugenol (ME), and ß-phellandrene (PHE) on the lipolytic activity and type II secretion system (T2SS) of Pseudomonas psychrophila KM02 (KM02) fish isolates grown in vitro and in fish model conditions. Spectrophotometric analysis with the p-NPP reagent showed inhibition of lipolysis from 11 to 46%. These results were confirmed by RT-qPCR, as the expression levels of lipA, lipB, and genes encoding T2SS were also considerably decreased. The supplementation of marinade with BPEO and TEO contributed to KM02 growth inhibition during vacuum packaging of salmon fillets relative to control samples. Whole-genome sequencing (WGS) provided insight into the spoilage potential of KM02, proving its importance as a spoilage microorganism whose metabolic activity should be inhibited to maintain the quality and safety of fresh fish in the food market.

Analysis of the Ability to Produce Pleasant Aromas on Sour Whey and Buttermilk By-Products by Mold Galactomyces geotrichum: Identification of Key Odorants.

Currently, there is a growing demand for flavorings, especially of natural origin. It is worth paying attention to the biotechnological processes of flavor production, characterized by simplicity, high efficiency and relatively low cost. In this study, we analyzed the ability of the Galac tomyces geotrichum mold to transform by-products of the dairy industry: sour whey and buttermilk to complex flavour mixtures with pleasant, honey-rose aroma. Furthermore, the aroma complexity of the fermentation product has been carefully identified applying a sensomic approach involving the use of gas chromatography-olfactometry (GC-O), gas chromatography-mass spectrometry (GC-MS) and stable isotope dilution assay (SIDA) to identify and quantify aroma compounds. Based on the calculation of odor activity value (OAV), 13 key aroma compounds were present in both tested variants. The highest OAVs were found for phenylacetaldehyde (honey-like) in the buttermilk variant (912) and 2-phenylethanol (rose-like) in the sour whey variant (524). High values of this indicator were also recorded for phenylacetaldehyde (319) and 3-methyl-1-butanol with a fruity aroma (149) in the sour whey culture. The other compounds identified are 3-methylbutanal (malty), 2,3-butanedione (cheesy), isovaleric acid (cheesy), 3-(methylthio)-propanal (boiled potato), butanoic acid (vinegar), (E)-2-nonenal (fatty), ethyl furaneol (burnt sugar), dimethyl trisulfide (cabbage), and acetic acid (vinegar).

In situ approaches show the limitation of the spoilage potential of Juniperus phoenicea L. essential oil against cold-tolerant Pseudomonas fluorescens KM24.

The present study aimed to elucidate the effect of subinhibitory concentrations (sub-MICs) of juniper essential oil (EO), α-pinene, and sabinene on the quorum-sensing (QS)-mediated proteolytic and lipolytic properties of Pseudomonas fluorescens KM24. These activities were verified under in situ conditions, in which sub-MICs of the agents altered the morphology of KM24 cells. RNA-Seq studies revealed key coding sequences (CDSs)/genes related to QS and the proteolytic/lipolytic activities of pseudomonads. In this work, all the examined agents decreased autoinducer synthesis and influenced the mRNA expression of the encoding acyltransferase genes lptA, lptD, and plsB. The highest reduction on the 3rd and 5th days of cultivation was observed for the genes lptD (-5.5 and -5.61, respectively) and lptA (-3.5 and -4.0, respectively) following treatment with EO. Inhibition of the lptA, lptD, and plsB genes by singular constituents of EO was on average, from -0.4 to -0.7. At 5 days of cultivation the profile of AHLs of the reference P. fluorescens KM24 strain consisted of 3-oxo-C14-HSL, 3-oxo-C6-HSL, C4-HSL, and N-[(RS)-3-hydroxybutyryl]-HSL, the concentrations of which were 0.570, 0.018, 3.744, and 0.554 µg ml-1, respectively. Independent of the incubation time, EO, α-pinene, and sabinene also suppressed the protease genes prlC (-1.5, -0.5, and -0.5, respectively) and ctpB (-1.5, -0.7, and -0.4, respectively). Lipolysis and transcription of the lipA/lipB genes were downregulated by the agents on average from -0.3 to -0.6. α-Pinene- and sabinene-rich juniper EO acts as an anti-quorum-sensing agent and can repress the spoilage phenotype of pseudomonads. KEY POINTS: Juniper EO, α-pinene, sabinene exhibited anti-QS potential toward KM24. RNA-Seq revealed key CDSs/genes related to QS/proteolytic/lipolytic activities of KM24. Agents at sub-MIC levels influenced the mRNA expression of QS/lipase/protease genes.

The Use of Sour and Sweet Whey in Producing Compositions with Pleasant Aromas Using the Mold Galactomyces geotrichum: Identification of Key Odorants.

Fermented products with a pleasant aroma and with strong honey, rose, and fruit odor notes were developed through the biotransformation of a medium containing sour or sweet whey with the addition of l-phenylalanine by the Galactomyces geotrichum mold. In order to obtain the strong honey-rose aroma, G. geotrichum strains were screened and fermentation conditions were optimized to achieve a preferable ratio (>1) of phenylacetaldehyde to 2-phenylethanol by the Ehrlich pathway. This allowed post-fermentation products with the ratio of concentrations of phenylacetaldehyde to 2-phenylethanol being 1.7:1. Additionally, the use of gas chromatography-olfactometry (GC-O) analysis and the calculation of odor activity values (OAVs) allowed 10 key odorants to be identified in post-fermentation products. The highest OAVs were found for phenylacetaldehyde with a honey odor in both sour and sweet whey cultures (3010 and 1776, respectively). In the variant with sour whey, the following compounds with the highest OAVs were 3-methyl-1-butanol (131), 3-(methylthio)-propanal (119), 3-methylbutanal (90), dimethyl trisulfide (71), 2,3-butanedione (37), and 2-phenylethanol (29). In the post-fermentation product with sweet whey, the following compounds with the highest OAVs were 3-(methylthio)-propanal (112), dimethyl trisulfide (69), and 2,3-butanedione (41).

Tarragon essential oil as a source of bioactive compounds with anti-quorum sensing and anti-proteolytic activity against Pseudomonas spp. isolated from fish - in vitro, in silico and in situ approaches.

The present study aimed to evaluate the anti-quorum sensing (anti-QS) and anti-proteolytic potentials of tarragon essential oil (TEO) and its major compounds against food-associated Pseudomonas spp. The activities were verified by in vitro, in silico and in situ approaches. In this work, methyl eugenol (ME)- and ß-phellandrene (ß-PH)-rich TEO was investigated. TEO at subMIC increased the percentage of saturated fatty acids in the bacterial membranes (from 7 to 22%) and exhibited anti-quorum sensing via decreasing the efficiency of QS autoinducer synthesis [3-oxo-C12-HSL (from 2.028 µg/mL to

Galactomyces geotrichum mold isolated from a traditional fried cottage cheese produced omega-3 fatty acids.

Thirty nine strains of Galactomyces geotrichum molds were isolated from a traditional fried cottage cheese and production of polyunsaturated fatty acids (PUFA) was assessed. Among them eleven strains produced an extracellular lipids enriched in n-6 and n-3 PUFA. The extracellular lipids produced by G. geotrichum strain 38 contained the highest amounts of total PUFA (24.3%), with the highest contribution of n-3 fatty acids (17.9%), where α-linolenic, eicosapentaenoic, docosapentaenoic and docosahexaenoic acids were the main contributors. To obtain maximal production of PUFA, composition of the medium consisted of 10 g/L rapeseed oil, 5 g/L yeast extract, 0.05 g/L K2HPO4, 0.17 g/L MgSO4, 0.015 g/L MnSO4, 0.015 g/L ZnSO4, 0.05 g/L FeSO4, and 10 mg/L vitamin B12. The optimal growth conditions at 30 °C involve: aeration at 1.5 vvm (volume of air per volume of broth per minute) at pH 6.5. The cheese produced under described conditions contained higher amount of n-3 PUFA (0.25 mg/g cheese) in comparison to control (0.01 mg/g). α-Linolenic acid predominated among n-3 fatty acids. Galactomyces geotrichum is a natural microflora of dairy products, and could be used to enrich food/cheese in deficient omega-3 lipids.

Production of Bioactive Compounds by Food Associated Galactomyces geotrichum 38, as Determined by Proteome Analysis.

Fried cottage cheese is a dairy product, popular in some parts of Poland. Proteomic analysis of a culture of the mold Galactomyces geotrichum 38 isolated from fried cottage cheese was performed using UHPLC/MS. From the proteins identified, we selected those involved in the biosynthesis of bioactive compounds and those useful in industry. In the G. geotrichum 38 culture, the production quantities of vitamin B2 (224 µg/L), ergosterol (54.63 mg/kg), and trehalose (0.91 g/L) were determined by HPLC. The identified proteins were also used to prepare a hypothetical fatty acid biosynthesis pathway, and the percentage of individual sphingolipids in the culture was determined. Sphingolipids are also bioactive compounds. During culturing of G. geotrichum 38, the percentage of three sphingolipids increased. The last step of the research was to prepare a model of fried cottage cheese. The mold G. geotrichum 38, used in the process of ripening fried cottage cheese, synthesized vitamin B2 and erogsterol, which influenced the nutritional value of the product.

Key Odorants of Lazur, a Polish Mold-Ripened Cheese.

Application of gas chromatography-olfactometry (GC-O) carried out on the volatile fraction isolated by solvent-assisted flavor evaporation (SAFE) and solid phase microextraction (SPME) from Lazur mold-ripened cheese revealed 17 odor-active compounds. The highest flavor dilution factor (FD) has been obtained for methanethiol (2048) with a burnt odor note and for 2(3)-methylbutanoic acid (2048) with a cheesy, pungent odor. Further quantitation of the 15 most aroma-active compounds allowed for calculation of their odor activity values (OAV). The highest OAVs were obtained for methanethiol (500), 3(2)-methylbutanoic acid (321), 3-(methylthio)propanal (210), 2,3-butanedione (65), dimethyl trisulfide (22), butanoic acid (20), 1-octen-3-ol (18), ( Z)-4-heptenal (14), dimethyl disulfide (14), dimethyl sulfide (13), phenylacetaldehyde (6), 2-ethyl-3,5-dimethylpyrazine (5), and acetic acid (4). An aroma recombination experiment showed slight differences in the perception of cheesy/sweaty and moldy/musty notes. To verify the influence of methyl ketones on the aroma profile of mold-ripened cheese, recombinant has been additionally supplemented with 2-pentanone, 2-heptanone, and 2-nonanone in concentrations determined in Lazur cheese. The aroma profile remained unchanged, which would suggest that methyl ketones, in this particular cheese, do not play a significant role in the formation of aroma.

Galactomyces geotrichum - moulds from dairy products with high biotechnological potential.

The article reviews the properties of the Galactomyces geotrichum species, the mould that is most important for the dairy industry. G. geotrichum mould has been isolated from milk, cheeses and alcoholic beverage. Its presence in food products makes it possible to obtain a characteristic aroma and taste, which corresponds to the needs and preferences of consumers. G. geotrichum plays an important role in ecology, where the mould is employed for the degradation of various hazardous substances and wastewater treatment. It has also been found to have potential for biofuel production. In addition to this, G. geotrichum can be applicable in two further major areas: agriculture and health protection.

Isolation of rhamnolipids-producing cultures from faeces: Influence of interspecies communication on the yield of rhamnolipid congeners.

The aim of this study was to evaluate the ability of bacterial cultures isolated from cattle, poultry or pig faeces and manure to produce rhamnolipids, as well as to investigate the influence of interspecies communication on possible quantitative differences in the production of rhamnolipid congeners. Initial screening methods (oil spreading, drop collapse, haemolytic activity and emulsification activity) showed that approximately 36% of the 51 isolated cultures exhibited the ability to produce biosurfactants. Subsequent studies using a selected culturable mixed culture (which included Enterococcus faecalis, Pseudomonas aeruginosa and Escherichia coli) revealed that only P. aeruginosa was able to produce this biosurfactant. HPLC-MS analysis showed that the surface active compounds were rhamnolipids. Further comparative studies confirmed that the total yield of rhamnolipids was notably higher in the bioreactor inoculated with the selected mixed culture (940.58±1.10mg/L) compared to the bioreactor inoculated with the axenic strain of P. aeruginosa (108.47±0.41mg/L). Twelve rhamnolipid congeners were identified during cultivation of the selected mixed culture, whereas six congeners were detected during cultivation of the sole axenic strain of P. aeruginosa. Furthermore, increased production of rhamnolipids was observed when the concentration of autoinducer molecules (AI-2) responsible for interspecies signaling increased, suggesting the influence of quorum-sensing communication on biosynthesis efficiency. This observation may be of importance for large-scale production of this biosurfactant, as it opens new possible solutions based on the use of mixed cultures or external addition of stimulating autoinducers.

Characterization of specific spoilage organisms (SSOs) in vacuum-packed ham by culture-plating techniques and MiSeq next-generation sequencing technologies.

BACKGROUND: Knowledge regarding microaerophilic and anaerobic specific spoilage organisms (SSOs) is crucial for an appropriate evaluation of vacuum-packed ham. The objective of this study was to characterize the SSO community in vacuum-packed ham by a culture-dependent technique and MiSeq next-generation sequencing (NGS) platform. The relation between changes among the SSO group in the ham and changes in sensory characteristics of the product was also assessed. RESULTS: In the study, conventional microbiological analyses were employed in order to establish the participation of several groups of microorganisms in the deterioration of vacuum-packed ham. The diversity of the SSO group in the product was further assessed with the use of MiSeq NGS technology. The bacteria identified in sliced cooked ham belonged mostly to four phyla, namely Actinobacteria, Proteobacteria, Firmicutes and Bacteroidetes. A temperature of 4 °C favoured the development of mesophilic and psychrophilic/psychrotrophic flora, mainly Lactobacillaceae, Enterobacteriaceae and Micrococcaceae families. A high ratio of Brochothrix thermosphacta species and new, cold-tolerant Clostridium spp. was also observed. The growth of these microorganisms facilitated changes in the pH value and organoleptic characteristics of the product. CONCLUSION: This study confirms that the combination of culturing and MiSeq NGS technology improves the microbial evaluation of food. © 2016 Society of Chemical Industry.

Gastrointestinal digested Sambucus nigra L. fruit extract protects in vitro cultured human colon cells against oxidative stress.

Elderberry (EDB) Sambucus nigra L. is one of the oldest medicinal plants which is useful for therapeutic and nutritional purposes due to a large amount of biologically active constituents, including compounds with a high antioxidant capacity. The present study focused on the antioxidant potential of the colon-available EDB fruit extract, derived from the artificial gastrointestinal tract, with regard to human colonic mucosa cells cultured in vitro. Despite the significant loss of EDB bioactive compounds due to the digestion process, the colon-digested extract was able to reduce the excessive intracellular ROS production (22%) and oxidative DNA damage (46%) in the colon cells at a dose of 1 mg of freeze-dried EDB powder/ml. Moreover, the colon-digested EDB extract inhibited oxidant-induced mutagenicity (26%) in the Salmonella typhimurium TA102 strain, as determined by the Ames test. In conclusion, the current in vitro study confirmed that the fruits of S. nigra are capable of protecting colonic cells against the detrimental effects of oxidative stress.

Role of gallic and p-coumaric acids in the AHL-dependent expression of flgA gene and in the process of biofilm formation in food-associated Pseudomonas fluorescens KM120.

BACKGROUND: In the process of Pseudomonas fluorescens biofilm formation, N-acyl-l-homoserine lactone (AHL)-mediated flagella synthesis plays a key role. Inhibition of AHL production may attenuate P. fluorescens biofilm on solid surfaces. This work validated the anti-biofilm properties of p-coumaric and gallic acids via the ability of phenolics to suppress AHL synthesis in P. fluorescens KM120. The dependence between synthesis of AHL molecules, expression of flagella gene (flgA) and the ability of biofilm formation by P. fluorescens KM120 on a stainless steel surface (type 304L) was also investigated. RESULTS: Research was carried out in a purpose-built flow cell device. Limitations on AHL synthesis in P. fluorescens KM120 were observed at concentrations of 120 and 240 µmol L(-1) of phenolic acids in medium. At such levels of gallic and p-coumaric acids the ability of P. fluorescens KM120 to synthesize 3-oxo-C6-homoserine lactone (HSL) was not observed. These concentrations caused decreased expression of flgA gene in P. fluorescens KM120. The changes in expression of AHL-dependent flgA gene significantly decreased the rate of microorganism colonization on the stainless steel surface. CONCLUSION: Phenolic acids are able to inhibit biofilm formation. The results obtained in the work may help to develop alternative techniques for anti-biofilm treatment in the food industry. © 2015 Society of Chemical Industry.

Investigation of the effectiveness of disinfectants against planktonic and biofilm forms of P. aeruginosa and E. faecalis cells using a compilation of cultivation, microscopic and flow cytometric techniques.

This study evaluated the effectiveness of selected disinfectants against bacterial cells within a biofilm using flow cytometry, the conventional total viable count test and scanning electron microscopy (SEM). A flow cytometric procedure based on measurement of the cellular redox potential (CRP) was demonstrated to have potential for the rapid evaluation of activity against biofilm and planktonic forms of microbes. Quaternary ammonium compound-based disinfectant (QACB) demonstrated a higher level of anti-microbial activity than a performic acid preparation (PAP), with mean CRP values against P. aeruginosa cells of 2 and 1.33 relative fluorescence units (RFU) vs 63.33 and 61.33 RFU for 8 and 24 h cultures respectively. Flow cytometric evaluation of the anti-biofilm activity demonstrated a higher efficacy of QACB compared to PAP for P. aeruginosa cells of 1 and 0.66 RFU vs 18.33 and 22.66 RFU for 8 and 24 h cultures respectively. SEM images of treated P. aeruginosa cells demonstrated disinfectant-specific effects on cell morphology.