Deciphering the core microbes and their interactions in spontaneous Baijiu fermentation: A comprehensive review.

The spontaneous Baijiu fermentation system harbors a complex microbiome that is highly dynamic in time and space and varies depending on the Jiuqu starters and environmental factors. The intricate microbiota presents in the fermentation environment is responsible for carrying out various reactions. These reactions necessitate the interaction among the core microbes to influence the community function, ultimately shaping the distinct Baijiu styles through the process of spontaneous fermentation. Numerous studies have been conducted to enhance our understanding of the diversity, succession, and function of microbial communities with the aim of improving fermentation manipulation. However, a comprehensive and critical assessment of the core microbes and their interaction remains one of the significant challenges in the Baijiu fermentation industry. This paper focuses on the fermentation properties of core microbes. We discuss the state of the art of microbial traceability, highlighting the crucial role of environmental and starter microbiota in the Baijiu brewing microbiome. Also, we discuss the various interactions between microbes in the Baijiu production system and propose a potential conceptual framework that involves constructing predictive network models to simplify and quantify microbial interactions using co-culture models. This approach offers effective strategies for understanding the core microbes and their interactions, thus beneficial for the management of microbiota and the regulation of interactions in Baijiu fermentation processes.

Enhancing bacterial cellulose production of Komagataeibacter nataicola through fermented coconut water by Saccharomyces cerevisiae: A metabonomics approach.

Nata de coco, an edible bacterial cellulose (BC) product, is a traditional dessert fermented in coconut water. Production of Nata de coco by Komagataeibacter nataicola is enhanced by pre-fermented coconut water, but its instability is a challenge. Here, BC production by K. nataicola Y19 was significantly improved by Saccharomyces cerevisiae 84-3 through shaping the metabolite profile of the coconut water. Different fermentation time with S. cerevisiae 84-3 resulted in distinct metabolite profiles and different promoting effect on BC yield. Compared to unfermented coconut water, coconut water fermented by S. cerevisiae 84-3 for 1d and 7d enhanced BC yield by 14.1-fold and 5.63-fold, respectively. Analysis between unfermented coconut water and 1d-fermented coconut water showed 129 significantly different metabolites, including organic acids, amino acids, nucleotides, and their derivatives. Prolonged fermentation for 7d changed levels of 155 metabolites belongs to organic acids, amino acids, nucleotides and their derivatives. Spearman correlation analysis further revealed that 17 metabolites were positively correlated with BC yield and 21 metabolites were negatively correlated with BC yield. These metabolites may affect energy metabolism, cell signaling, membrane integrity, and BC production by K. nataicola Y19. The further verification experiment gave the view that BC yield was not only closely related to the types of metabolites but also the concentration of metabolites. This study provides a novel theoretical framework for a highly efficient BC fermentation system utilizing stable fermented coconut water mediums.

Multiple metabolite profiles uncover remarkable bioactive compounds and metabolic characteristics of noni fruit (Morinda citrifolia L.) at various stages of ripeness.

This study aimed to investigate the changes in physicochemical properties, bioactive compounds, and metabolic characteristics of noni fruit at different ripeness levels. The results showed that there were significant differences in physicochemical properties. HPLC analysis was conducted, revealing succinic acid, scopoletin, deacetylasperulosidic acid, and asperulosidic acid were key bioactive compounds as the fruit ripened. Additionally, 4 differentbiomarkers (isocitric acid, 4,4-thiodiphenol, lobaric acid, and octocrylene), identified using 1HNMR and LC-IT-TOF-MS, were found to have a VIP value over 1. The results from HS-GC-IMS demonstrated noteworthy that 14 volatile compounds were identified as highly discriminative features during fruit ripening. Furthermore, correlation analysis showed that different ripeness had significant effects on bioactive components and functional activities, e.g., the inhibition rate of enzyme and E. coli of noni fruit with different ripeness exceeded 90% at the last stage. This study contributes new insights into the effective utilization of bioactive ingredients in noni fruit.

Insights into the antibacterial effectiveness of linalool against Shigella flexneri on pork surface: Changes in bacterial growth and pork quality.

Shigella flexneri has the ability to contaminate pork and cause foodborne diseases. This study aimed to examine the effectiveness of linalool (a natural preservative) against S. flexneri and explore its potential application in contaminated pork. The results showed that linalool was capable of damaging the cell membrane and binding to the DNA of S. flexneri, and inhibiting biofilm formation and disrupting mature biofilms. The antibacterial effectiveness of linalool on the surface of pork was further demonstrated by analyzing the physicochemical properties of the pork (i.e., weight loss rate, pH value, color index, and TVB-N value) and its protein profiles. Linalool did not completely kill S. flexneri in pork at minimum bactericidal concentration (MBC) concentration and its antibacterial effect of linalool was stronger during the initial stage of storage. During storage, linalool influenced the abundance of specific proteins in the pork, particularly those involved in pathways related to fat metabolism. These findings offer novel insights into the antibacterial efficacy of linalool and its underlying mechanism in pork.

Exploring the biotic and abiotic drivers influencing nata de coco production by Komagataeibacter nataicola in pre-fermented coconut water.

In China and Southeast Asia, pre-fermented coconut water is commonly used for the production of nata de coco, a jelly-like fermented food that consists of bacterial cellulose (BC). The inherent natural fermentation process of coconut water introduces uncontrollable variables, which can lead to unstable yields during BC production. This study involved the collection of spontaneously pre-fermented coconut water over a five-month production cycle. The aim was to evaluate the microbiota and metabolite profile, as well as determine its impact on BC synthesis by Komagataeibacter nataicola. Significant variations in the microbial community structure and metabolite profile of pre-fermented coconut water were observed across different production months, these variations had significant effects on BC synthesis by K. nataicola. A total of 52 different bacterial genera and 32 different fungal genera were identified as potential biotic factors that can influence BC production. Additionally, several abiotic factors, including lactate (VIP = 4.92), mannitol (VIP = 4.22), ethanol (VIP = 2.67), and ascorbate (VIP = 1.61), were found to be potential driving forces affecting BC synthesis by K. nataicola. Upon further analysis, the correlation network indicated that 14 biotic factors had a significant contribution to BC production in three strains of K. nataicola. These factors included 8 bacterial genera, such as Limosilactobacillus and Lactiplantibacillus, and 6 fungal genera, such as Meyerozyma and Ogataea. The abiotic factors lactate, mannitol, and ethanol showed a positive correlation with the BC yield. This study provides significant insights into controlling the fermentation processes of pre-fermented coconut water in industrial settings.

Lactiplantibacillus plantarum and Saccharomyces cerevisiae-Fermented Coconut Water Alleviates Dextran Sodium Sulfate-Induced Enteritis in Wenchang Chicken: A Gut Microbiota and Metabolomic Approach.

In order to investigate the potential mechanisms of probiotic-fermented coconut water in treating enteritis, this study conducted a comprehensive analysis of the effects of probiotic intervention on the recovery from Dextran Sodium Sulfate-induced acute enteritis in Wenchang chicks. The analysis encompassed the assessment of growth performance, serum indicators, intestinal tissue structure, and metagenomic and metabolomic profiles of cecal contents in 60 Wenchang chicks subjected to intervention. This approach aimed to elucidate the impact of probiotic intervention on the recovery process from acute enteritis at both the genetic and metabolic levels in the avian model. The results revealed that intervention with Saccharomyces cerevisiae Y301 improved the growth rate of chicks. and intervention with Lactiplantibacillus plantarum MS2c regulated the glycerophospholipid metabolism pathway and reshaped the gut microbiota structure in modeling chicks with acute enteritis, reducing the abundance of potentially pathogenic bacteria from the Alistipes and increasing the abundance of potentially beneficial species from the Christensenellaceae. This intervention resulted in the production of specific gut metabolites, including Gentamicin C and polymyxin B2, recognized for their therapeutic effects on acute enteritis. The combined intervention of S. cerevisiae Y301 and L. plantarum MS2c not only enhanced growth performance but also mitigated intestinal wall damage and increased the abundance of gut metabolites such as gentamicin C and polymyxin B2, thereby mitigating symptoms of enteritis. Furthermore, this combined intervention reduced the levels of serum immune markers, including IL-10, IL-6, TNF-α, IFN-γ, and D-lactic acid, thus mitigating intestinal epithelial cell damage and promoting acute enteritis recovery. This study provides crucial insights into the mechanisms of action of probiotics and probiotic-fermented coconut water in acute enteritis recovery, offering new perspectives for sustainable farming practices for Wenchang chicken.

pH-responsive chitosan-based film containing oregano essential oil and black rice bran anthocyanin for preserving pork and monitoring freshness.

This study developed an intelligent and antibacterial packaging film using a chitosan matrix embedding oregano essential oil (OEO) and black rice bran anthocyanin (BRBA). Herein, OEO and BRBA were immobilized into the chitosan matrix through noncovalent bonds and uniformly distributed in the films. The chitosan-OEO-BRBAⅡ film exhibited excellent mechanical, antibacterial, antioxidant, and UV-vis light barrier properties, and sensitive and rapid response to pH/NH3. Furthermore, fresh pork was coated with the film for monitoring the freshness and preservation efficiency at 4 °C for 12 days. The film effectively improved the quality indices of pork, including the sensory index, total viable counts, pH, TVB-N value, and color of pork during the storage at 4 °C. The film reduced the abundance of spoilage bacteria related to stress tolerance, pathogenicity, and biofilm formation in the pork. Their odorous volatiles appeared later and were less than those in the untreated group.

Response of microbial community assembly and succession pattern to abiotic factors during the second round of light-flavor Baijiu fermentation.

The regulation of microbial communities is an important strategy for fermentation management. Dynamic microbiota during the Baijiu fermentation is shaped by a variety of abiotic factors. Therefore, this study aims to investigate the effects of the microbiota of the fermentation starter Daqu on the microbial assembly and the interaction of microbiota succession and abiotic factors during the second round (Ercha) of light-flavor Baijiu fermentation. The results revealed that Streptomyces, Bacillus, Lactobacillus, and Staphylococcus were the dominant bacterial genera in the initial fermentation, while Lactobacillus was dominant during the middle and later stages. Pichia and Saccharomycopsis were the dominant fungal genera during the whole fermentation process. A total of 54 volatile compounds were identified during the fermentation, among which 15 compounds, mainly including ethyl acetate, diethyl azelate, ethyl 2-hydroxyisocaproate, 3-furaldehyde, and ethylidene diacetate, were identified as important flavor metabolites. The SourceTracker software revealed that Daqu contributed 52.3 % of the bacterial community and 38.6 % of the fungal community to the fermentation. Ethanol, moisture, and pH were the major factors regulating the succession of dominant bacteria and fungi during the fermentation. The microbial succession and co-occurrence pattern driven by abiotic factors played a crucial role in shaping flavor profiles. These results provide guidance for controlling the fermentation process by optimizing operational parameters or bioaugmentation with specific microbes.

Integrated multi-omics approaches to understand microbiome assembly in Jiuqu, a mixed-culture starter.

The use of Jiuqu as a saccharifying and fermenting starter in the production of fermented foods is a very old biotechnological process that can be traced back to ancient times. Jiuqu harbors a hub of microbial communities, in which prokaryotes and eukaryotes cohabit, interact, and communicate. However, the spontaneous fermentation based on empirical processing hardly guarantees the stable assembly of the microbiome and a standardized quality of Jiuqu. This review describes the state of the art, limitations, and challenges towards the application of traditional and omics-based technology to study the Jiuqu microbiome and highlights the need for integrating meta-omics data. In addition, we review the varieties of Jiuqu and their production processes, with particular attention to factors shaping the microbiota of Jiuqu. Then, the potentials of integrated omics approaches used in Jiuqu research are examined in order to understand the assembly of the microbiome and improve the quality of the products. A variety of different approaches, including molecular and mass spectrometry-based techniques, have led to scientific advances in the analysis of the complex ecosystem of Jiuqu. To date, the extensive research on Jiuqu has mainly focused on the microbial community diversity, flavor profiles, and biochemical characteristics. An integrative approach to large-scale omics datasets and cultivated microbiota has great potential for understanding the interrelation of the Jiuqu microbiome. Further research on the Jiuqu microbiome may explain the inherent property of compositional stability and stable performance of a complex microbiota coping with environmental perturbations and provide important insights to reconstruct synthetic microbiota and develop modern intelligent manufacturing procedures for Jiuqu.

Insights into the bacterial, fungal, and phage communities and volatile profiles in different types of Daqu.

Daqu has a rich and diverse microbiota, giving them a suitable biotope for phages. However, the absolute abundances of bacteria and fungi, as well as the phage community characteristics in Daqu, remain unclear. In this study, the microbiota absolute abundance, indigenous phage composition and function, and volatile compound profiles of high-temperature Daqu (HTDQ), medium-temperature Daqu (MTDQ), and low-temperature Daqu (LTDQ) were investigated. Absolute microbiota quantification revealed that there were significant variations in microbial composition and absolute abundance across three types of Daqu. The absolute abundances of the top 30 bacterial genera in LTDQ, MTDQ, and HTDQ were 6.0 × 105, 5.3 × 104, 1.4 × 105 copies/ng DNA, while the top 30 fungal genera had 8.5 × 107, 2.1 × 106, and 6.2 × 105 copies/ng DNA, respectively. LTDQ were enriched in Pantoea, Staphylococcus, and Saccharomycopsis; MTDQ were dominated by Saccharopolyspora, Staphylococcus, Saccharomycopsis, and Aspergillus; HTDQ were dominated by Saccharopolyspora, Bacillus, Byssochlamys, and Saccharomycopsis. Volatile profile analysis revealed that LTDQ, MTDQ, and HTDQ comprised 68, 68, and 75 compounds, respectively, with 39 compounds shared by the three types. Fourteen volatile compounds were identified as highly discriminative features among three types of Daqu, which were closely related to Saccharopolyspora, Thermoactinomyces, Kroppenstedtia, Byssochlamys, and Thermomyces. Metaviromics indicated that Genomoviridae, Mimiviridae and Siphoviridae, and Parvoviridae were the dominant viruses in LTDQ, MTDQ, and HTDQ, respectively. The hosts of phages in Daqu mainly included Lactobacillus, Enterobacter, Pantoea, Bacillus, Pediococcus, and Staphylococcus. Phages may prey on numerous microbes living on Daqu via pathways such as genetic information processing, signaling and cellular processes, and replication and repair. This study highlights the use of absolute quantification to avoid misjudgment of differential taxa in comparative microbiome analysis and provides new insights into the phage community and function in Daqu.

Unraveling the microbial compositions, metabolic functions, and antibacterial properties of Huangshui, a byproduct of Baijiu fermentation.

Huangshui (HS) is a byproduct of Chinese Baijiu production that has tremendous potential to become a high-value product in the food industry. This study comprehensively analyzed the commonalities and differences in physicochemical properties, microbial structures, metabolomes, flavor profiles, and antibacterial properties among sources of HS. The HS samples were extracted from three different pits in three different workshops within an industrial Baijiu distillery. Lactobacillus was the dominant genus of bacteria in all of them, while Kazachstania, Aspergillus, and Pichia were the dominant genera of fungi. A total of 45 volatile compounds, including ethyl hexanoate, hexanoic acid, and 1-butanol, were detected in the HS by headspace solid-phase microextraction coupled with gas chromatography mass spectrometry, suggesting HS could be used to flavor liquor. Untargeted metabolomic analysis yielded 224 nonvolatile metabolites that were common in all HS samples. These metabolites included organic acids, carbohydrates, benzenoids, nucleic acids, and fatty acids, which are mainly involved in amino acid and sugar metabolism. While the compositions of the microbiome and metabolome among HS samples were affected by workshop factors, there were essentially no differences in the bactericidal properties of the HS samples among workshops. HS exhibited potent antibacterial activities against Escherichia coli and Staphylococcus aureus, and HS was effective in inactivating mature biofilms formed on polystyrene microplates, as confirmed by scanning electron micrographs. Our results provided a theoretical basis to increase the economic value of HS for applications in the food industry.

Contrasting summer versus winter dynamic microbial communities and their environmental driving factors in the solid-state saccharification process of Fuyu-flavor Baijiu.

The seasonal spontaneous fermentation depends on the assembly of specific microbiota. However, a knowledge gap remains regarding the seasonal dynamics of microbial community and functionality. This study aimed to investigate the differences in microbial community dynamics during the saccharification process of Fuyu-flavor Baijiu in winter versus summer by employing high-throughput sequencing. It also explored the driving effects of environmental variables on the microbial succession. The environmental temperature of saccharification in summer (29 ± 1 °C) was strikingly different in winter (14 ± 1 °C), leading to a longer incubation time in winter fermentation. The dominant bacterial genera displayed in the early stage of the summer saccharification process were Staphylococcus, unclassified Enterobacteriaceae, and Weissella, whereas the dominant genus was Weissella during the middle stage and Lactobacillus during the late stage of saccharification. In contrast, unclassified Enterobacteriaceae and Weissella were dominant genera throughout the winter saccharification process. In the fungal community, Rhizopus was the dominant genus in summer and winter saccharification. Principal coordinate analysis revealed that the changes in the environmental temperature in winter versus summer had distinct impacts on the microbial community structure. Also, temperature was the key factor driving the succession of bacterial community, whereas pH was the main factor driving the succession of dominant fungi. Predictive gene functionality of bacterial and fungal communities was computed using the Phylogenetic Investigation of Communities by Reconstruction of Unobserved States 2 against Kyoto Encyclopedia of Genes and Genomes (KEGG) database and MetaCyc database, respectively. Correlation networks were constructed to better understand the relationships between dominant microbial genera and physicochemical factors during the saccharification process. These results provided new insights into the Baijiu fermentation technology and allowed a theoretical basis for exploring effective management strategies regarding traditional fermentation in response to complex seasonal factors.

Multidimensional exploration of essential oils generated via eight oregano cultivars: Compositions, chemodiversities, and antibacterial capacities.

Numerous species of Origanum (Lamiaceae) have been widely used as spices to extend the shelf life of foods. Essential oils extracted from this genus have attracted much attention owing to their potential applications as bactericides. Here, we evaluated the chemical compositions of eight oregano essential oils (OEOs) using gas chromatography-mass spectrometry and assessed their antibacterial activities. The chemical compositions of OEOs were affected by the cultivar factor, and seven common compounds, including carvacrol, were identified among eight OEOs. Partial least squares discriminant analysis enabled the distinction of three groups among these OEOs, as characterized by the proportions of carvacrol, thymol, and sesquiterpenes. OEOs effectively inhibited Escherichia coli and Staphylococcus aureus with varying antibacterial activities. Spearman correlation network highlighted core antibacterial contributors in the chemical profiles of OEOs. Our results revealed that the bacteriostatic effects of OEOs could be explained by core compounds and their synergistic effects.

Deciphering the Shifts in Microbial Community Diversity From Material Pretreatment to Saccharification Process of Fuyu-Flavor Baijiu.

The microbiota of the pretreatment phase is crucial to the assembly of the microbial community in the saccharification of fuyu-flavor baijiu. This study investigates the shifts in microbial community diversity from the pretreatment of raw materials to the end of saccharification. High-throughput sequencing reveals that Lactobacillus, Weissella, and Bacillus in the bacterial community and Rhizopus, Candida, Pichia, and Aspergillus in the fungal community are predominant during raw material pretreatment and saccharification processes. Also, 11 bacterial genera, including Bacillus, Lactobacillus, Leuconostoc, Weissella, Lactococcus, and Acetobacter, and eight yeast genera, including Candida, Pichia, Saccharomyces, and Wickerhamomyces, were isolated from the initial saccharification stage by culture-dependent approaches. Sourcetracker analysis indicates that the cooling grains and rice husks were the main contributors to the bacterial community composition of the saccharification process, and Qu was the main contributor to the shaping of the fungal community structure during the saccharification process. Abundance variation of the predictive functional profiles of microbial communities encoding for key enzymes involved in pyruvate metabolism, starch and sucrose metabolism, and glycolysis/gluconeogenesis during the pretreatment and saccharification phases were inferred by PICRUSt2 analysis. The results of this study will be utilized to produce consistently high-quality fuyu-flavor baijiu via better controlling the shaping of microbial community structures during the pretreatment and fermentation processes.

Comparison of Nutritional Compositions and Essential Oil Profiles of Different Parts of a Dill and Two Fennel Cultivars.

Fennel and dill are widely used as food additives owing to their various biological activities, such as antioxidants, antimicrobials, food-preservatives, and seasoning capacities. Herein, the nutritional composition and essential oil (EO) chemical profiles of fruits, umbels, stalks, and roots from one dill and two fennel cultivars were evaluated. The fruits had the highest content of crude protein (≥15%), crude fat (≥8%), and phosphorus (≥0.5%), and exhibited the highest total energy (≥20 MJ/kg) and EO yield (≥2%). Moreover, estragole (86.56% in Fdf), anethole (71.17% in Fhf), fenchone (16.74% in Fhf), limonene (50.19% in Agf), and carvone (42.41% in Agf) were the main components of the EOs generated from the fruits. The chemical profiles of EOs in the roots were significantly different from those of the aerial parts of the fennel and dill; thus, the roots and aerial parts could be distinguished based on myristicin (Variable Importance in Projection (VIP) = 1.90399) and apiol (VIP = 1.85922). The EO components of the aerial parts varied remarkably, and the chemical markers for differentiating these three cultivars were anethole (VIP = 1.36571), estragole (VIP = 1.30292), and carvone (VIP = 1.11947). Overall, our results provide a noteworthy chemical basis for further development of fennel and dill, especially as food additives.

Occurrence of disinfectant-resistant bacteria in a fresh-cut vegetables processing facility and their role in protecting Salmonella enteritidis.

Chemical disinfectants are widely used to control foodborne pathogen contamination in fresh-cut vegetables (FVs) processing facilities. In this study, we investigated the disinfectant-resistant bacteria in a FVs processing facility and evaluate the effects of these bacteria on Salmonella enteritidis biofilm formation and disinfectant resistance. The disinfectant-resistance profiles were determined using 0.02% sodium hypochlorite (NaClO), 0.2% benzalkonium bromide (BAB) and 2% hydrogen peroxide (H2O2) solutions. The results showed the high occurrence of disinfectant resistant bacteria in the FVs processing environment, especially in the clean area. All isolates showed planktonic susceptibility to H2O2 and BAB, while the Gram-positive isolates were specifically resistant to NaClO. Isolates with biofilm-forming ability showed resistance to tested disinfectants. Disinfectant resistance of S. enteritidis was not significantly enhanced in most of the mixed-species biofilms, except for Bacillus paramycoides B5 which not only increased the biomass but also enhanced the survival ability of the Salmonella under NaClO treatment. Increased biomass and compact biofilm structures were observed in mixed-species biofilms by scanning electron microscopy (SEM). This study provides new insights into the disinfectant-resistant bacteria from food processing facilities and highlights their relevance for foodborne pathogen contamination.

Composition and Metabolic Functions of the Microbiome in Fermented Grain during Light-Flavor Baijiu Fermentation.

The metabolism and accumulation of flavor compounds in Chinese Baijiu are driven by microbiota succession and their inter-related metabolic processes. Changes in the microbiome composition during Baijiu production have been examined previously; however, the respective metabolic functions remain unclear. Using shotgun metagenomic sequencing and metabolomics, we examined the microbial and metabolic characteristics during light-flavor Baijiu fermentation to assess the correlations between microorganisms and their potential functions. During fermentation, the bacterial abundance increased from 58.2% to 97.65%, and fermentation resulted in the accumulation of various metabolites, among which alcohols and esters were the most abundant. Correlation analyses revealed that the levels of major metabolites were positively correlated with bacterial abundance but negatively with that of fungi. Gene annotation showed that the Lactobacillus species contained key enzyme genes for carbohydrate metabolism and contributed to the entire fermentation process. Lichtheimia ramosa, Saccharomycopsis fibuligera, Bacillus licheniformis, Saccharomyces cerevisiae, and Pichia kudriavzevii play major roles in starch degradation and ethanol production. A link was established between the composition and metabolic functions of the microbiota involved in Baijiu fermentation, which helps elucidate microbial and metabolic patterns of fermentation and provides insights into the potential optimization of Baijiu production.

Ferulic Acid Inactivates Shigella flexneri through Cell Membrane Destructieon, Biofilm Retardation, and Altered Gene Expression.

Antibiotic resistance and capacity for biofilm formation of Shigella flexneri render previous prevention and control strategies minimally effective. Ferulic acid (FA) has been demonstrated to be useful due to its application in foods as an alternative natural preservative. However, information regarding the S. flexneri phenotype and molecular responses to FA exposure is limited. The present study investigated the effects of FA on S. flexneri planktonic growth and biofilm formation. The results demonstrated that the cell membrane of S. flexneri in planktonic growth mode exhibited irreversible destruction after FA exposure, as characterized by decreased cell viability, leakage of cytoplasmic constituents, accelerated adenosine triphosphate (ATP) consumption, cell membrane depolarization, and cellular morphological changes. FA significantly inhibited S. flexneri adhesion and biofilm formation at a working concentration (1/8 MIC) that almost did not inhibit planktonic growth. Transcriptomics profiling showed that the exposure to a subinhibitory concentration of FA dramatically altered gene expression in the S. flexneri biofilm, as a total of 169 differentially expressed genes (DEGs) were upregulated and 533 DEGs were downregulated, compared to the intact biofilm. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis showed that the DEGs were mainly involved in pathways of ribosomes, ABC transporters, and the citrate cycle. Furthermore, we show that FA altered the transcription of S. flexneri genes associated with adhesion, transcriptional regulation, and the synthesis and transport of extracellular polymeric substances that contribute to biofilm formation. These data provide novel insights into S. flexneri behavioral responses to FA exposure and suggest that FA could effectively constrain S. flexneri and its biofilm formation.

Effect of thyme essential oil against Bacillus cereus planktonic growth and biofilm formation.

The objective of this study was to determine the effect of thyme essential oil (TEO) on the planktonic growth and biofilm formation of Bacillus cereus (B. cereus). GC-MS analysis of TEO allowed the detection of 13 compounds, and the major constituents were p-cymene (29.7%), thymol (23.73%), γ-terpinene (16.21%), and 1,8-cineole (9.74%). TEO exhibited a minimum inhibitory concentration (MIC) value against planktonic B. cereus of 0.25 mg/mL. The potent effect of TEO to inhibit the growth of planktonic B. cereus was due to cell membrane damage, as evidenced by reduced cell viability, protein changes, decreased intracellular ATP concentration, increased extracellular ATP concentration and cell membrane depolarization, and cellular morphological changes. In addition, TEO exerted a significant inhibitory effect on B. cereus biofilm formation, as confirmed by environmental scanning electron microscopic images. These findings suggested that TEO has the potential to be developed as a natural food additive to control foodborne contamination associated with B. cereus and its biofilm.

The specific effect of gallic acid on Escherichia coli biofilm formation by regulating pgaABCD genes expression.

Escherichia coli (E. coli) is associated with an array of health-threatening contaminations, some of which are related to biofilm states. The pgaABCD-encoded poly-beta-1,6-N-acetyl-D-glucosamine (PGA) polymer plays an important role in biofilm formation. This study was conducted to determine the inhibitory effect of gallic acid (GA) against E. coli biofilm formation. Minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) values of GA against planktonic E. coli were 0.5 and 4 mg/mL, and minimal biofilm inhibitory concentration and minimal biofilm eradication concentration values of GA against E. coli in biofilms were 2 and 8 mg/mL, respectively. Quantitative crystal violet staining of biofilms and ESEM images clearly indicate that GA effectively, dose-dependently inhibited biofilm formation. CFU counting and confocal laser scanning microscopy measurements showed that GA significantly reduced viable bacteria in the biofilm. The contents of polysaccharide slime, protein, and DNA in the E. coli biofilm also decreased. qRT-PCR data showed that at the sub-MIC level of GA (0.25 mg/mL) and expression of pgaABC genes was downregulated, while pgaD gene expression was upregulated. The sub-MBC level of GA (2 mg/mL) significantly suppressed the pgaABCD genes. Our results altogether demonstrate that GA inhibited viable bacteria and E. coli biofilm formation, marking a novel approach to the prevention and treatment of biofilm-related infections in the food industry.