Probiotics and biofilm interaction in aquaculture for sustainable food security: A review and bibliometric analysis.

Aquaculture is one of the most significant food sources from the prehistoric period. As aquaculture intensifies globally, the prevalence and outbreaks of various pathogenic microorganisms cause fish disease and heavy mortality, leading to a drastic reduction in yield and substantial economic loss. With the modernization of the aquaculture system, a new challenge regarding biofilms or bacterial microenvironments arises worldwide, which facilitates pathogenic microorganisms to survive under unfavorable environmental conditions and withstand various treatments, especially antibiotics and other chemical disinfectants. However, we focus on the mechanistic association between those microbes which mainly form biofilm and probiotics in one of the major food production systems, aquaculture. In recent years, probiotics and their derivatives have attracted much attention in the fisheries sector to combat the survival strategy of pathogenic bacteria. Apart from this, Bibliometric analysis provides a comprehensive overview of the published literature, highlighting key research themes, emerging topics, and areas that require further investigation. This information is valuable for researchers, policymakers, and stakeholders in determining research priorities and allocating resources effectively.

Comprehensive metagenomic analysis of stress-resistant and -sensitive Listeria monocytogenes.

Listeria monocytogenes is a pathogenic bacterium which can live in adverse environments (low pH, high salinity, and low temperature). Even though there are various whole genome sequencing (WGS) data on L. monocytogenes, investigations on genetic differences between stress-resistant and -sensitive L. monocytogenes grown under stress environments have been not fully examined. This study aims to investigate and compare genetic characteristics between stress-resistant and -sensitive L. monocytogenes using whole genome sequencing (WGS). A total of 47 L. monocytogenes strains (43 stress-resistant and 4 stress-sensitive) were selected based on the stress-resistance tests under pH 3, 5% salt concentration, and 1 °C. The sequencing library for WGS was prepared and sequenced using an Illumina MiSeq. Genetic characteristics of two different L. monocytogenes groups were examined to analyze the pangenome, functionality, virulence, antibiotic resistance, core, and unique genes. The functionality of unique genes in the stress-resistant L. monocytogenes was distinct compared to the stress-sensitive L. monocytogenes, such as carbohydrate and nucleotide transport and metabolism. The lisR virulence gene was detected more in the stress-resistant L. monocytogenes than in the stress-sensitive group. Five stress-resistant L. monocytogenes strains possessed tet(M) antibiotic resistance gene. This is the first study suggesting that deep genomic characteristics of L. monocytogenes may have different resistance level under stress conditions. This new insight will aid in understanding the genetic relationship between stress-resistant and -sensitive L. monocytogenes strains isolated from diverse resources. KEY POINTS: • Whole genomes of L. monocytogenes isolated from three different sources were analyzed. • Differences in two L. monocytogenes groups were identified in functionality, virulence, and antibiotic resistance genes. • This study first examines the association between resistances and whole genomes of stress-resistant and -sensitive L. monocytogenes.

Innovative nonthermal technologies for inactivation of emerging foodborne viruses.

Various foodborne viruses have been associated with human health during the last decade, causing gastroenteritis and a huge economic burden worldwide. Furthermore, the emergence of new variants of infectious viruses is growing continuously. Inactivation of foodborne viruses in the food industry is a formidable task because although viruses cannot grow in foods, they can survive in the food matrix during food processing and storage environments. Conventional inactivation methods pose various drawbacks, necessitating more effective and environmentally friendly techniques for controlling foodborne viruses during food production and processing. Various inactivation approaches for controlling foodborne viruses have been attempted in the food industry. However, some traditionally used techniques, such as disinfectant-based or heat treatment, are not always efficient. Nonthermal techniques are considered a new platform for effective and safe treatment to inactivate foodborne viruses. This review focuses on foodborne viruses commonly associated with human gastroenteritis, including newly emerged viruses, such as sapovirus and Aichi virus. It also investigates the use of chemical and nonthermal physical treatments as effective technologies to inactivate foodborne viruses.

Current and future interventions for improving poultry health and poultry food safety and security: A comprehensive review.

Poultry is thriving across the globe. Chicken meat is the most preferred poultry worldwide, and its popularity is increasing. However, poultry also threatens human hygiene, especially as a fomite of infectious diseases caused by the major foodborne pathogens (Campylobacter, Salmonella, and Listeria). Preventing pathogenic bacterial biofilm is crucial in the chicken industry due to increasing food safety hazards caused by recurring contamination and the rapid degradation of meat, as well as the increased resistance of bacteria to cleaning and disinfection procedures commonly used in chicken processing plants. To address this, various innovative and promising strategies to combat bacterial resistance and biofilm are emerging to improve food safety and quality and extend shelf-life. In particular, natural compounds are attractive because of their potential antimicrobial activities. Natural compounds can also boost the immune system and improve poultry health and performance. In addition to phytochemicals, bacteriophages, nanoparticles, coatings, enzymes, and probiotics represent unique and environmentally friendly strategies in the poultry processing industry to prevent foodborne pathogens from reaching the consumer. Lactoferrin, bacteriocin, antimicrobial peptides, cell-free supernatants, and biosurfactants are also of considerable interest for their prospective application as natural antimicrobials for improving the safety of raw poultry meat. This review aims to describe the feasibility of these proposed strategies and provide an overview of recent published evidences to control microorganisms in the poultry industry, considering the human health, food safety, and economic aspects of poultry production.

Insights into antibiofilm mechanisms of phytochemicals: Prospects in the food industry.

The recalcitrance of microbial aggregation or biofilm in the food industry underpins the emerging antimicrobial resistance among foodborne pathogens, exacerbating the phenomena of food spoilage, processing and safety management failure, and the prevalence of foodborne illnesses. The challenges of growing tolerance to current chemical and disinfectant-based antibiofilm strategies have driven the urgency in finding a less vulnerable to bacterial resistance, effective alternative antibiofilm agent. To address these issues, various novel strategies are suggested in current days to combat bacterial biofilm. Among the innovative approaches, phytochemicals have already demonstrated their excellent performance in preventing biofilm formation and bactericidal actions against resident bacteria within biofilms. However, the diverse group of phytochemicals and their different modes of action become a barrier to applying them against specific pathogenic biofilm-formers. This phenomenon mandates the need to elucidate the multi-mechanistic actions of phytochemicals to design an effective novel antibiofilm strategy. Therefore, this review critically illustrates the structure - activity relationship, functional sites of actions, and target molecules of diverse phytochemicals regarding multiple major antibiofilm mechanisms and reversal mechanisms of antimicrobial resistance. The implementation of the in-depth knowledge will hopefully aid future studies for developing phytochemical-based next-generation antimicrobials.

Efficacy of ficin and peroxyacetic acid against Salmonella enterica serovar Thompson biofilm on plastic, eggshell, and chicken skin.

Salmonella is the leading cause of zoonotic foodborne illnesses worldwide and a prevalent threat to the poultry industry. For controlling contamination, the use of chemical sanitizers in combination with biological compounds (e.g., enzymes) offers a solution to reduce the chemical residues. The current study investigated the biofilm reduction effects of a food-grade enzyme-ficin-and a common sanitizer-peroxyacetic acid (PAA)-against an emerging pathogen, Salmonella enterica ser. Thompson, on plastic, eggshell, and chicken skin surfaces. Results showed that PAA could kill S. Thompson, but ficin cannot. Maximum biofilm reduction was 3.7 log CFU/cm2 from plastic after individual treatment with PAA. However, sequential treatment of ficin and PAA led to biofilm reductions of 3.2, 5.0, and 6.5 log CFU/cm2 from chicken skin, eggshell, and plastic, respectively. Fourier-transform infrared spectroscopy and microscopic analysis confirmed that ficin increased PAA action, causing biofilm matrix destruction. Moreover, the quality of the food surfaces was only altered by 12.5 U/mL ficin and was not altered by PAA. This combined use of enzyme and sanitizer solved major safety issues and proved promising against S. Thompson-associated contaminations in poultry and poultry processing lines.

Combination treatment of peroxyacetic acid or lactic acid with UV-C to control Salmonella Enteritidis biofilms on food contact surface and chicken skin.

The risk of salmonellosis is expected to increase with the rise in the consumption of poultry meat. The aim of this study was to investigate the combination treatment of peroxyacetic acid (PAA) or lactic acid (LA) with UV-C against Salmonella Enteritidis biofilms formed on food contact surface (stainless steel [SS], silicone rubber [SR], and ultra-high molecular weight polyethylene [UHMWPE]) and chicken skin. The biofilm on food contact surface and chicken skin was significantly decreased (P < 0.05) by combination treatment of PAA or LA with UV-C. Combination treatment of PAA (50-500 µg/mL) with UV-C (5 and 10 min) reduced 3.10-6.41 log CFU/cm2 and LA (0.5-2.0%) with UV-C (5 and 10 min) reduced 3.35-6.41 log CFU/cm2 of S. Enteritidis biofilms on food contact surface. Salmonella Enteritidis biofilms on chicken skin was reduced around 2 log CFU/g with minor quality changes in color and texture by combination treatment of PAA (500 µg/mL) or LA (2.0%) with UV-C (10 min). Additional reduction occurred on SS and UHMWPE by PAA or LA with UV-C, while only LA with UV-C caused additional reduction on chicken skin. Also, it was visualized that the biofilm on food contact surface and chicken skin was removed through field emission scanning electron microscopy (FESEM) and death of cells constituting the biofilm was confirmed through confocal laser scanning microscopy (CLSM). These results indicating that the combination treatment of PAA or LA with UV-C could be used for S. Enteritidis biofilm control strategy in poultry industry.

Microbial and chemical composition of Cheddar cheese supplemented with prebiotics from pasteurized milk to aging.

Microbial and chemical properties of cheese is crucial in the dairy industry to understand their effects on cheese quality. Microorganisms within this fat, protein, and water matrix are largely responsible for physiochemical characteristics and associated quality. Prebiotics can be used as an energy source for lactic acid bacteria in cheese by altering the microbial community and provide the potential for value-added foods, with a more stable probiotic population. This research focuses on the addition of fructooligosaccharides (FOS) or inulin to the Cheddar cheese-making process to evaluate the effects on microbial and physicochemical composition changes. Laboratory-scale Cheddar cheese produced in 2 replicates was supplemented with 0 (control), 0.5, 1.0, and 2.0% (wt/wt) of FOS or inulin using 18 L of commercially pasteurized milk. A total of 210 samples (15 samples per replicate of each treatment) were collected from cheese-making procedure and aging period. Analysis for each sample were performed for quantitative analysis of chemical and microbial composition. The prevalence of lactic acid bacteria (log cfu/g) in Cheddar cheese supplemented with FOS (6.34 ± 0.11 and 8.99 ± 0.46; ± standard deviation) or inulin (6.02 ± 0.79 and 9.08 ± 1.00) was significantly higher than the control (5.84 ± 0.27 and 8.48 ± 0.06) in whey and curd, respectively. Fructooligosaccharides supplemented cheeses showed similar chemical properties to the control cheese, whereas inulin-supplemented cheeses exhibited a significantly higher moisture content than FOS and the control groups. Streptococcus and Lactococcus were predominant in all cheeses and 2% inulin and 2% FOS-supplemented cheeses possessed significant amounts of nonstarter lactic acid bacteria found to be an unidentified group of Lactobacillaceae, which emerged after 90 d of aging. In conclusion, this study demonstrates that prebiotic supplementation of Cheddar cheese results in differing microbial and chemical characteristics.

Current and future perspectives for controlling Vibrio biofilms in the seafood industry: a comprehensive review.

The contamination of seafood with Vibrio species can have severe repercussions in the seafood industry. Vibrio species can form mature biofilms and persist on the surface of several seafoods such as crabs, oysters, mussels, and shrimp, for extended duration. Several conventional approaches have been employed to inhibit the growth of planktonic cells and prevent the formation of Vibrio biofilms. Since Vibrio biofilms are mostly resistant to these control measures, novel alternative methods need to be urgently developed. In this review, we propose environmentally friendly approaches to suppress Vibrio biofilm formation using a hypothesized mechanism of action.

Shifts of microbiota during cheese production: impact on production and quality.

The high-throughput DNA sequencing (HTS) method is used to identify microbes in cheese and their potential functional properties. The technique can be applied to the microbiota of the cheese processing environment, raw milk, curd, whey, and starter cultures, and be used to improve the quality, safety, and other physicochemical properties of the final product. The HTS method is also utilized to study the microbiota shift of different types of cheeses during processing, as the composition and functional properties of the microbiome provide unique characteristics to different cheeses. Although there are several reviews that focused on microbiota of various types of cheeses, this review focuses on evaluating the microbiota shift of different types of cheese production and highlights key bacteria in each step of the processing as well as microbiota of various types of cheeses. KEY POINTS: • High-throughput sequencing can be applied to identify microbiota in cheese. • Microbiota in cheese is changed during making process and aging. • Starter culture plays an important role to establish microbiota in cheese.

The application of bacteriophage to control Cronobacter sakazakii planktonic and biofilm growth in infant formula milk.

The goal was to identify the biofilm-forming ability of Cronobacter sakazakii on surfaces of stainless steel (SS) and silicone rubber (SR) in contact with infant formula milk. Two representative bacteriophages (PBES04 and PBES19) were used to control the growth of C. sakazakii as well as its biofilm forming ability on either SS or SR surfaces. Bacterial growth was confirmed at 20 °C when PBES04 and PBES19 were used, whereas C. sakazakii was not normally detected in infant formula milk treated with both bacteriophages for 6 h. In an additional biofilm reduction experiment, the biofilm on SS or SR surfaces were reduced by 3.07 and 1.92 log CFU cm-2, respectively after PBES04 treatment, and 3.06 and 2.14 log CFU cm-2, respectively, after PBES19 treatment. These results demonstrate that bacteriophages can be effective in inactivating C. sakazakii in biofilms which could potentially increase food safety in commercial facilities.

Comprehensive molecular, probiotic, and quorum-sensing characterization of anti-listerial lactic acid bacteria, and application as bioprotective in a food (milk) model.

Listeria monocytogenes is a major foodborne pathogen that adversely affects the food industry. In this study, 6 anti-listerial lactic acid bacteria (LAB) isolates were screened. These anti-listerial LAB isolates were identified via 16S rRNA gene sequencing and analyzed via repetitive extragenic palindromic-PCR. Probiotic assessment of these isolates, comprising an evaluation of the antibiotic susceptibility, tolerance to lysozyme, simulated gastric and intestinal juices, and gut conditions (low pH, bile salts, and 0.4% phenol), was carried out. Most of the isolates were resistant to streptomycin, vancomycin, gentamycin, kanamycin, and ciprofloxacin. All of the isolates were negative for virulence genes, including agg, ccf, cylA, cylB, cylLL, cylLS, cylM, esp, and gelE, and hemolytic activity. Furthermore, autoinducer-2 (a quorum-sensing molecule) was detected and quantified via HPLC with fluorescence detection after derivatization with 2,3-diaminonaphthalene. Metabolites profiles of the Lactobacillus sakei D.7 and Lactobacillus plantarum I.60 were observed and presented various organic acids linked with antibacterial activity. Moreover, freeze-dried cell-free supernatants from Lb. sakei (55 mg/mL) and Lb. plantarum (40 mg/mL) showed different minimum effective concentration (MEC) against L. monocytogenes in the food model (whole milk). In summary, these anti-listerial LAB isolates do not pose a risk to consumer health, are eco-friendly, and may be promising candidates for future use as bioprotective cultures and new probiotics to control contamination by L. monocytogenes in the food and dairy industries.

Promising strategies to control persistent enemies: Some new technologies to combat biofilm in the food industry-A review.

Biofilm is an advanced form of protection that allows bacterial cells to withstand adverse environmental conditions. The complex structure of biofilm results from genetic-related mechanisms besides other factors such as bacterial morphology or substratum properties. Inhibition of biofilm formation of harmful bacteria (spoilage and pathogenic bacteria) is a critical task in the food industry because of the enhanced resistance of biofilm bacteria to stress, such as cleaning and disinfection methods traditionally used in food processing plants, and the increased food safety risks threatening consumer health caused by recurrent contamination and rapid deterioration of food by biofilm cells. Therefore, it is urgent to find methods and strategies for effectively combating bacterial biofilm formation and eradicating mature biofilms. Innovative and promising approaches to control bacteria and their biofilms are emerging. These new approaches range from methods based on natural ingredients to the use of nanoparticles. This literature review aims to describe the efficacy of these strategies and provide an overview of recent promising biofilm control technologies in the food processing sector.

Design of PCR assays to specifically detect and identify 37 Lactobacillus species in a single 96 well plate.

BACKGROUND: Lactobacillus species are used as probiotics and play an important role in fermented food production. However, use of 16S rRNA gene sequences as standard markers for the differentiation of Lactobacillus species offers a very limited scope, as several species of Lactobacillus share similar 16S rRNA gene sequences. In this study, we developed a rapid and accurate method based on comparative genomic analysis for the identification of 37 Lactobacillus species that are commonly used in probiotics and fermented foods. RESULTS: To select species-specific sequences or genes, a total of 180 Lactobacillus genome sequences were compared using Python scripts. In 14 out of 37 species, species-specific sequences could not be found due to the similarity of the 16S-23S rRNA gene. Selected unique genes were obtained using comparative genomic analysis and all genes were confirmed to be specific for 52,478,804 genomes via in silico analysis; they were found not to be strain-specific, but to exist in all strains of the same species. Species-specific primer pairs were designed from the selected 16S-23S rRNA gene sequences or unique genes of species. The specificity of the species-specific primer pairs was confirmed using reference strains, and the accuracy and efficiency of the polymerase chain reaction (PCR) with the standard curve were confirmed. The PCR method developed in this study is able to accurately differentiate species that were not distinguishable using the 16S rRNA gene alone. This PCR assays were designed to detect and identify 37 Lactobacillus species. The developed method was then applied in the monitoring of 19 probiotics and 12 dairy products. The applied tests confirmed that the species detected in 17 products matched those indicated on their labels, whereas the remaining products contained species other than those appearing on the label. CONCLUSIONS: The method developed in this study is able to rapidly and accurately distinguish different species of Lactobacillus, and can be used to monitor specific Lactobacillus species in foods such as probiotics and dairy products.

Prevalence of extended-spectrum ß-lactamases in the local farm environment and livestock: challenges to mitigate antimicrobial resistance.

The effectiveness of antibiotics has been challenged by the increasing frequency of antimicrobial resistance (AR), which has emerged as a major threat to global health. Despite the negative impact of AR on health, there are few effective strategies for reducing AR in food-producing animals. Of the antimicrobial resistant microorganisms (ARMs), extended-spectrum ß-lactamases (ESBLs)-producing Enterobacteriaceae are an emerging global threat due to their increasing prevalence in livestock, even in animals raised without antibiotics. Many reviews are available for the positive selection of AR associated with antibiotic use in livestock, but less attention has been given to how other factors including soil, water, manure, wildlife, and farm workers, are associated with the emergence of ESBL-producing bacteria. Understanding of antibiotic resistance genes and bacteria transfer at the interfaces of livestock and other potential reservoirs will provide insights for the development of mitigation strategies for AR.

Intake of Watermelon or Its Byproducts Alters Glucose Metabolism, the Microbiome, and Hepatic Proinflammatory Metabolites in High-Fat-Fed Male C57BL/6 J Mice.

BACKGROUND: Watermelon intake has demonstrated effects on blood pressure regulation along with other health benefits. OBJECTIVE: We hypothesized that intake of whole watermelon and products made from watermelon rind (WR) and watermelon skin (WS) would remediate metabolic complications in C57BL/6 J male mice fed a diet modeling a Western-style diet. METHODS: Ten-week-old male C57BL/6 J mice were provided either a low-fat (LF) diet [10% fat (by energy), 8% sucrose (by energy) and no added cholesterol], a high-fat (HF) diet [45% fat (by energy), 20% kcal sucrose (by energy), and 1% (w/w) cholesterol], or an HF diet plus WS, WR, or watermelon flesh (WF) for 10 wk. Dried WF was provided at 8% of total energy (equivalent to 2 servings/d) and watermelon skin and rind were added at 2.25% (w/w, dry weight of additives) of diet. Animals were provided experimental diets ad libitum. Body weights, food intake, and glucose tolerance were determined. Serum insulin, inflammatory markers, microbiome, and the relative hepatic concentrations of 709 biochemicals were measured postmortem. RESULTS: The final body weight of the LF control group was significantly lower than that of the HF-fed control group (32.8 ± 0.9 g compared with 43.0 ± 1.7 g, P ≤ 0.05). Mice in treatment groups fed HF supplemented with watermelon products had final body weights similar to those of the HF-fed control mice. Serum insulin concentrations were reduced by ∼40% in mice fed an HF diet with WR supplementation compared with mice fed an HF diet alone (P ≤ 0.05). Depending on the individual species or group, microbiome populations changed significantly. Supplementation with WF resulted in a return to the basal hepatic concentrations of monohydroxy fatty acids and eicosanoids observed in LF-fed mice (P ≤ 0.05). CONCLUSIONS: In obese male mice, supplementation with each of the watermelon products to an HF diet improved fasting blood glucose, circulating serum insulin concentrations, and changes in hepatic metabolite accumulation. At a modest level of supplementation to an HF diet, fiber-rich additives made from WR and WS further improved glucose metabolism and energy efficiency and shifted the microbiome composition.

Assessment of overall microbial community shift during Cheddar cheese production from raw milk to aging.

Cheese is a fermented dairy product that is made from animal milk and is considered to be a healthy food due to its available nutrients and potential probiotic characteristics. Since the microbes in the cheese matrix directly contribute to the quality and physicochemical properties of cheese, it is important to understand the microbial properties of cheese. In this study, Cheddar cheeses produced on three different dates at the Arbuthnot Dairy Center at Oregon State University were collected to determine the microbial community structure. A total of 773,821 sequencing reads and 271 amplicon sequence variants (ASVs) were acquired from 108 samples. Streptococcus and Lactococcus were observed as the most abundant ASVs in the cheese, which were used as the starter lactic acid bacteria (SLAB). Escherichia coli was detected in the raw milk; however, it was not detected after inoculating with SLAB. According to an alpha diversity analysis, SLAB inoculation decreased the microbial richness by inhibiting the growth of other bacteria present in the milk. A beta diversity analysis showed that microbial communities before the addition of SLAB clustered together, as did the samples from cheese making and aging. Non-starter lactic acid bacteria (NSLAB) were detected 15 weeks into aging for the June 6th and June 26th produced cheeses, and 17 weeks into aging for the cheese produced on April 26th. These NSLAB were identified as an unidentified group of Lactobacillaceae. This study characterizes the changes in the Cheddar cheese microbiome over the course of production from raw milk to a 6-month-aged final product. KEY POINTS: • 271 ASVs were acquired from cheese production from raw milk to 6-month aging. • Addition of SLAB changed the microbial diversity during Cheddar cheese making procedure. • NSLAB were detected more than 15 weeks after aging. Graphical Abstract.

Antibacterial and antibiofilm mechanism of eugenol against antibiotic resistance Vibrio parahaemolyticus.

The objective of this study was to investigate the antibacterial and antibiofilm activity of eugenol against V. parahaemolyticus planktonic and biofilm cells and the involved mechanisms as well. Atime-kill assay, a biofilm formation assay on the surface of crab shells, an assay to determine the reduction of virulence using eugenol at different concentrations, energy-filtered transmission electron microscope (EF-TEM), field emission scanning electron microscopy (FE-SEM), confocal laser scanning microscope (CLSM) and high-performance liquid chromatography (HPLC) were performed to evaluate the antibacterial and antibiofilm activity of eugenol. The results indicated that different concentrations of eugenol (0.1-0.6%) significantly reduced biofilm formation, metabolic activities, and secretion of extracellular polysaccharide (EPS), with effective antibacterial effect. Eugenol at 0.4% effectively eradicated the biofilms formed by clinical and environmental V. parahaemolyticus on crab surface by more than 4.5 and 4 log CFU/cm2, respectively. At 0.6% concentration, the reduction rates of metabolic activities for ATCC27969 and NIFS29 were 79% and 68%, respectively. Whereas, the reduction rates of EPS for ATCC27969 and NIFS29 were 78% and 71%, respectively. On visual evaluation, significant results were observed for biofilm reduction, live/dead cell detection, and quorum sensing (QS). This study demonstrated that eugenol can be used to control V. parahaemolyticus biofilms and biofilm-related infections and can be employed for the protection of seafood.

Microbial communities of a variety of cheeses and comparison between core and rind region of cheeses.

Understanding the microbial community of cheese is important in the dairy industry, as the microbiota contributes to the safety, quality, and physicochemical and sensory properties of cheese. In this study, the microbial compositions of different cheeses (Cheddar, provolone, and Swiss cheese) and cheese locations (core, rind, and mixed) collected from the Arbuthnot Dairy Center at Oregon State University were analyzed using 16S rRNA gene amplicon sequencing with the Illumina MiSeq platform (Illumina, San Diego, CA). A total of 225 operational taxonomic units were identified from the 4,675,187 sequencing reads generated. Streptococcus was observed to be the most abundant organism in provolone (72 to 85%) and Swiss (60 to 67%), whereas Lactococcus spp. were found to dominate Cheddar cheese (27 to 76%). Species richness varied significantly by cheese. According to alpha diversity analysis, porter-soaked Cheddar cheese exhibited the highest microbial richness, whereas smoked provolone cheese showed the lowest. Rind regions of each cheese changed color through smoking and soaking for the beverage process. In addition, the microbial diversity of the rind region was higher than the core region because smoking and soaking processes directly contacted the rind region of each cheese. The microbial communities of the samples clustered by cheese, indicated that, within a given type of cheese, microbial compositions were very similar. Moreover, 34 operational taxonomic units were identified as biomarkers for different types of cheese through the linear discriminant analysis effect size method. Last, both carbohydrate and AA metabolites comprised more than 40% of the total functional annotated genes from 9 varieties of cheese samples. This study provides insight into the microbial composition of different types of cheese, as well as various locations within a cheese, which is applicable to its safety and sensory quality.

Salmonella Cold Stress Response: Mechanisms and Occurrence in Foods.

Since bacteria in foods often encounter various cold environments during food processing, such as chilling, cold chain distribution, and cold storage, lower temperatures can become a major stress environment for foodborne pathogens. Bacterial responses in stressful environments have been considered in the past, but now the importance of stress responses at the molecular level is becoming recognized. Documenting how bacterial changes occur at the molecular level may help to achieve the in-depth understanding of stress responses, to predict microbial fate when they encounter cold temperatures, and to design and develop more effective strategies to control pathogens in food for ensuring food safety. Microorganisms differ in responding to a sudden downshift in temperature and this, in turn, impacts their metabolic processes and can cause various structural modifications. In this review, the fundamental aspects of bacterial cold stress responses focused on cell membrane modification, DNA supercoiling modification, transcriptional and translational responses, cold-induced protein synthesis including CspA, CsdA, NusA, DnaA, RecA, RbfA, PNPase, KsgA, SrmB, trigger factors, and initiation factors are discussed. In this context, specific Salmonella responses to cold temperature including growth, injury, and survival and their physiological and genetic responses to cold environments with a focus on cross-protection, different gene expression levels, and virulence factors will be discussed.