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The effects of ultra-high pressure (UHP) pretreatment (50-250 MPa) on the fish curing were studied. UHP increased the overall volatile compound concentration of cured fish. Among 50-250 MPa five treatment groups, 150 MPa UHP group exhibited the highest total free amino acid content (294.34 mg/100 g) with that of the control group being 92.39 mg/100 g. The activity of cathepsin L was increased under 50-200 MPa UHP treatment (62.28-58.15 U/L), compared with that in the control group (53.80 U/L). UHP treatment resulted in a significant increase in small molecule compounds, especially the amino acid dipeptides and ATP metabolic products. Under UHP treatments, the bacterial phyla Actinobacteriota (1.04-5.25 %), Bacteroidota (0.20-4.47 %), and Deinococcota (0.00-0.05 %) exhibited an increased abundance, and they promoted taste and flavor formation. Our results indicated that UHP is a promising pretreatment method to improve taste and flavour in cured fish by affecting the microorganisms, cathepsin, and proteins., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

To enhance the value of surimi, efforts have been made to develop a fermentation method with lactic acid bacteria (LAB) to proteolyze fish protein. However, fermenting unheated surimi poses a spoilage risk due to its high bacterial content. Surimi heat treatment can prevent spoilage, but gel formation induced by heating introduces another technical issue: it hinders uniform fermentation. Thus, this study aims to observe the proteolysis and enhance the functionality of seafood product through lactic acid fermentation of kamaboko, a heated surimi. Upon analyzing the kamaboko fermented with Lactobacillus helveticus JCM1004, we observed that LAB produced protease, resulting in the degradation of myosin heavy chain and actin during fermentation. Lactic acid fermentation significantly augmented the peptide content of kamaboko, subsequently elevating the angiotensin Ⅰ-converting enzyme (ACE) inhibitory activity in 200-fold diluted extract of fermented kamaboko to approximately 70% and higher. Notably, our investigation revealed that proteolysis was confined to the surface of kamaboko, as evidenced by SDS-PAGE analysis. This observation implies that the surface area of kamaboko influences the ACE inhibitory activity. Through a comparative analysis of various bacterial strains, we demonstrated that the increase in ACE inhibitory activity is contingent on the protease generated by LAB. These results suggest that LAB-mediated proteolysis of fish proteins liberates bioactive peptides, thereby manifesting in the ACE inhibitory activity. In summary, this study underscores that the fermentation of kamaboko employing proteolytic LAB holds promise in the development of novel functional seafood products.

This study explored the dynamics of anchovy sauce fermentation and investigated how the raw material form and the use of starter cultures affect bacterial and metabolite profiles. Using a comprehensive approach, we examined the fermentation process using anchovies in two forms (whole and ground) and three different starter cultures. The use of ground anchovies resulted in an accelerated fermentation process for anchovy sauce; however, the increased diversity of bacterial phylotypes and altered accumulation of biogenic amines were observed. Inoculation of starter cultures resulted in a shift from spontaneous to controlled fermentation, highlighting their ability to regulate bacterial communities. Despite a slightly reduced fermentation rate, inoculation with Tetragenococcus halophilus was shown to be a potent method for reducing biogenic amines and affecting metabolite profiles. As the industry strives to balance fermentation speed and quality, our research could provide insights for improving the efficiency, safety, and quality of anchovy sauce production., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Numerous Pseudomonas species can infect aquatic animals, such as farmed rainbow trout, sea trout, sea bass, and sea bream, by causing disease or stress reactions. In aquaculture facilities, a number of Pseudomonas species have been isolated and identified as the main pathogens. The present study describes the characterization of 18 Pseudomonas strains, isolated from fish products using shotgun proteomics. The bacterial proteomes obtained were further analyzed to identify the main functional pathway proteins involved. In addition, this study revealed the presence of 1015 non-redundant peptides related to virulence factors. An additional 25 species-specific peptides were identified as putative Pseudomonas spp. biomarkers. The results constitute the largest dataset, described thus far for the rapid identification and characterization of Pseudomonas species present in edible fish; furthermore, these data can provide the basis for further research into the development of new therapies against these harmful pathogens., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Single starter can hardly improve the volatile flavor of fermented fish surimi. In this study, the changes of volatile compounds (VCs) and microbial composition during cooperative fermentation of Latilactobacillus sakei and Pediococcus acidilactici were studied by headspace solid-phase microextraction gas chromatography-mass spectrometry and 16S rRNA gene high-throughput sequencing. During cooperative fermentation, most VCs and the abundance of Latilactobacillus and Lactococcus significantly increased, while Pediococcus, Acinetobacter, and Macrococcus obviously decreased. After evaluation of correlation and abundance of each genus, Latilactobacillus and Lactococcus possessed the highest influence on the formation of volatile flavor during cooperative fermentation. Compared with the natural fermentation, cooperative fermentation with starters significantly enhanced most of pleasant core VCs (odor activity value≥1), but inhibited the production of trimethylamine and methanethiol, mainly resulting from the absolutely highest influence of Latilactobacillus. Cooperative fermentation of starters is an effective method to improve the volatile flavor in the fermented tilapia surimi., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Hui Huang reports was provided by National Key Research and Development Program of China. Hui Huang reports was provided by Earmarked fund for CARS. Shengjun Chen reports was provided by Science and Technology Program of Guangzhou of China. Jun Li reports was provided by Key Platform Project of Guangdong Provincial Department of Education. Yongqiang Zhao reports was provided by Central Public-interest Scientific Institution Basal Research Fund, CAFS. Chunsheng Li reports was provided by Central Public-interest Scientific Institution Basal Research Fund, South China Sea Fisheries Research Institute, CAFS. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Cured Spanish mackerel has a promising market owing to its nutritious nature as well as ease of transportation and preservation. However, the nutritional and flavor formation mechanism of Spanish mackerel after curing and drying is unclear. To overcome this problem, the effects of different processing conditions on the free amino acid, microbial community, and flavor of Spanish mackerel were explored. Staphylococcus and Cobetia are the main microorganisms in cured mackerel and are closely associated with the formation of their quality. Compared with fresh mackerel, cured mackerel contains increased levels of protein, fat, and chloride, contributing to its distinctive flavor. The contents of free amino acids in the BA64 group were substantially higher than those in other groups, particularly the contents of threonine, glycine, and tyrosine. These findings will contribute to the development of high-quality cured Spanish mackerel products and cured aquatic products., Competing Interests: Declaration of competing interest None, (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Single starter can hardly elevate the gel property of fermented freshwater fish sausage. In this work, in order to improve the physical properties of tilapia sausage, two newly isolated strains of lactic acid bacteria (LAB), Latilactobacillus sakei and Pediococcus acidilactici were used for cooperative fermentation of tilapia sausage, followed by the revelation of their formation mechanisms during cooperative fermentation and their improvement mechanisms after comparison with natural fermentation. Both strains, especially L. sakei possessed good growth, acidification ability, and salt tolerance. The gel strength, hardness, springiness, chewiness, whiteness, acidification, and total plate count significantly elevated during cooperative fermentation with starters. Pediococcus, Acinetobacter, and Macrococcus were abundant before fermentation, while Latilactobacillus quickly occupied the dominant position after fermentation for 18-45 h with the relative abundance over 51.5 %. The influence of each genus on the physical properties was calculated through the time-dimension and group-dimension correlation networks. The results suggested that the increase of Latilactobacillus due to the good growth and metabolism of L. sakei contributed the most to the formation and improvement of gel strength, texture properties, color, acidification, and food safety of tilapia sausage after cooperative fermentation. This study provides a novel analysis method to quantitatively evaluate the microbial contribution on the changes of various properties. The cooperative fermentation of LAB can be used for tilapia sausage fermentation to improve its physical properties., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The study aimed to isolate and characterize lactic acid bacteria from various traditional fermented fish products from North East India, including Xindol, Hentak, and Ngari, which hold significant dietary importance for the indigenous tribes. Additionally, the study sought to examine their untargeted metabolomic profiles. A total of 43 strains of Bacillus, Priestia, Staphylococcus, Pediococcus, and Lactiplantibacillus were isolated, characterized by 16 S rRNA gene and tested for probiotic properties. Five strains passed pH and bile salt tests with strain dependent antimicrobial activity, which exhibited moderate autoaggregation and hydrophobicity properties. Lactiplantibacillus plantarum MKTJ24 exhibited the highest hydrophobicity (42 %), which was further confirmed by adhesion assay in HT-29 cell lines (100 %). Lactiplantibacillus plantarum MKTJ24 treatment in LPS-stimulated HT-29 cells up-regulated expression of mucin genes compared to LPS-treated cells. Treatment of RAW 264.7 cells with Lactiplantibacillus plantarum MKTJ24 decreased LPS-induced reactive oxygen species (ROS) and nitric oxide (NO) productions. Further, genome analysis of Lactiplantibacillus plantarum MKTJ24 revealed the presence of several probiotic markers and immunomodulatory genes. The genome was found to harbor plantaricin operon involved in bacteriocin production. A pangenome analysis using all the publicly available L. plantarum genomes specifically isolated from fermented fish products identified 120 unique genes in Lactiplantibacillus plantarum MKTJ24. Metabolomic analysis indicated dominance of ascorbic acids, pentafluropropionate, cyclopropaneacetic acid, florobenzylamine, and furanone in Xindol. This study suggests that Lactiplantibacillus plantarum MKTJ24 has potential probiotic and immunomodulatory properties that could be used in processing traditional fermented fish products on an industrial scale to improve their quality and enhance functional properties., Competing Interests: Declaration of Competing Interest None., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

To investigate the mechanisms of flavor formation in dry-fermented tilapia sausages, the volatiles, bacterial community, and lipid composition during fermentation were analyzed using gas chromatography-ion mobility spectrometry, 16S high throughput sequencing, and ultra-performance liquid chromatography-mass spectrometer. Pediococcus pentosaceus, Staphylococcus xylosus, and Staphylococcus carnosus became dominant bacteria during the fermentation. A total of 66 volatiles and 293 lipids (48 differential lipids) were identified. PC and PE content decreased. Aldehyde and 1-octen-3-ol content decreased. Most esters and ketones content increased during fermentation. Six metabolic pathways associated with differential lipids were identified by enrichment analysis. Glycerophospholipid metabolism was the main metabolic pathway. Correlation analysis revealed that PC and PE were precursors for volatiles, including PC 16:0/18:2 and PE 18:0/22:6. The dominant bacteria facilitate the hydrolysis of PC and PE, leading to the formation of esters and ketones. This study provides a theoretical basis for the targeted regulation of fermented sausage flavors., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microbial metabolism is important for the unique flavor formation of Mei yu, a kind of traditional Chinese fermented fish pieces. However, the interactive relationship between microorganisms and flavor components during fermentation is still unclear. In this study, electronic nose and headspace-solid-phase microextraction-gas chromatography-mass spectrometry analysis were performed to identify flavor components in Mei yu during the fermentation, and the absolute microbial quantification was conducted to identify the diversity and succession of microbial communities. During fermentation, there was an increase in the types of volatile compounds. Alcohols, aldehydes, aromatics and esters were the main flavor compounds and significantly increased in Mei yu, while hydrocarbon and aldehydes significantly decreased. The absolute abundances of Lactobacillus, Lactococcus and Weissella increased significantly after 3 days' fermentation, which were closely associated with the productions of 1-nonanol, 2-methoxy-4-vinylphenol, guaiacol, ethyl palmitate and ethyl caprylate that might though pathways related to fatty acid biosynthesis and amino acid metabolism. However, these genera were negatively correlated with the production of indole. Additionally, the total volatile basic nitrogen (TVB-N) levels of Mei yu fermented during 3 days were within the limits of 25 mg TVB-N/100 g fish, with the contents of free amino acids and lipoxygenase activities were significant lower than that of 4 days' fermentation. In view of food safety and flavor, it suggested that the natural fermented Mei yu at room temperature should be controlled within 3 days. This study highlights the application of absolute quantification to microbiome analysis in traditional fermented Mei yu and provides new insights into the roles of microorganisms in flavor formation during fermentation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Lactic acid bacteria (LAB) were isolated from naturally fermented foods of India, viz., sidra, a dried fish product; kinema, a naturally fermented sticky soybean food; and dahi, a naturally fermented milk product. Five strains of LAB, based on 16S rRNA gene sequence, were identified: Lactococcus lactis FS2 (from sidra), Lc. lactis C2D (dahi), Lc. lactis SP2C4 (kinema), Lactiplantibacillus plantarum DHCU70 (=Lactobacillus plantarum) (from dahi), and Lactiplantibacillus plantarum KP1 (kinema). The PICRUSt2 software, a bioinformatic tool, was applied to infer the raw sequences obtained from LAB strains mapped against KEGG database for predictive functionality. Functional features of LAB strains showed genes associated with metabolism (36.47%), environmental information processing (31.42%), genetic information processing (9.83%), and the unclassified (22.28%). KEGG database also showed abundant genes related to predictive membrane transport (29.25%) and carbohydrate metabolism (11.91%). This study may help in understanding the health-promoting benefits of the culturable LAB strains in fermented foods., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

As a traditional Thai condiment, Pla-ra is used to add flavor and richness to dishes. Nine treatment combinations of Pla-ra formulations created from 3 types of fish (Mor fish, Kradee fish, and Mor + Kradee fish) and 4 different carbohydrate sources (none, rice bran, roasted rice, and rice bran─roasted rice mixture) were studied through a 12 month fermentation period (1, 3, 5, 7, 8, 9, 10, 11, and 12 months). 16S rRNA Next Generation Sequencing (NGS) and LC-MS/MS techniques were used to analyze the microbial diversity and identify taste-enhancing peptides. Descriptive sensory analysis was performed on the extracts of the 108 Pla-ra samples mixed in a model broth. Koku perception and saltiness-enhancing attributes were clearly perceived and dominant in all samples, even though glutamyl peptides, including γ-Glu-Val-Gly, were found at subthreshold levels. The samples from mixed fish and Mor fish fermented with roasted ground rice and rice bran for 12 months had the most typical Pla-ra odors and tastes and had high taste-enhancing activities. NGS analysis revealed the presence of bacteria containing a large number of protease and aminopeptidase genes in the samples. Bacillus spp., Gallicola spp., and Proteiniclasticum spp. correlated well with the generation of glutamyl and arginyl peptides and typical odors in the samples. These results confirmed the typical sensory quality of Pla-ra depended on protein sources, carbohydrate sources, and bacteria communities. Further optimization of the microbial composition found could lead to the development of starter cultures to control and promote flavor development in fermented fish products.

Fishery products are one of the main human nutritional sources, and due to the consumption increase, the quality of the derived products may be modified, during catching, technological processing, and storage. Detection and identification of pathogenic and spoilage microorganisms in fishery products is needed because the first may be involved in human diseases, while the second is responsible of significant economic losses. In this sense, liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) method and computational analysis of MS data are useful tools for characterizing and identifying different microorganisms and to develop promising strategies for food science investigations. Moreover, in the past decade, metaproteomic methodologies have progressed for the study of microorganisms isolated from their natural samples and independently of the culture restrictions. Metaproteomics enables assessment of proteins and pathways from individual members of the consortium. Metaproteomics can provide a detailed understanding of which organisms occupy specific metabolic niches, how they interact, and how they utilize nutrients, and these insights can be obtained directly from environmental samples.According to that, the sample preparation of the fishery product, the LC-ESI-MS/MS dedicated method, and the MS data analysis were described in the present chapter to obtain the metaproteomic analysis of the respective microbiomes or microbial communities., (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

Abstract: Histamine-forming bacteria (HFB) were isolated from the 70 salted fish samples bought from town markets of Guangdong Province of south China. In addition, the histamine-forming ability of HFB was analyzed. There were 31 strains of HFB isolated from 36 salted fish pickled overnight. They were identified as six bacteria species: Vibrio alginolyticus, Vibrio rumoiensis, Staphylococcus saprophyticus, Staphylococcus xylosus, Lactococcus lactis, and Morganella morganii. The rate of confirmation of V. alginolyticus was highest (23 of 31), exceeding 200 mg/kg histamine. In particular, M. morganii produced a histamine amount that exceeded 2,000 mg/kg, although it was only one isolate in this study. In addition, five bacteria species of HFB were isolated from 34 dried salted fish. Among them, S. saprophyticus was dominant in the dried salted fish but produced histamines below 200 mg/kg. However, Enterobacter aerogenes from dried salted fish formed a histamine amount exceeding 200 mg/kg. The study showed that the dominant strain of HFB was different in two kinds of salted fish. Both kinds of salted fish contained HFB whose histamine-forming capacity exceeded 200 mg/kg. As a result, the safety of salted fish should be of concern, especially salted fish pickled overnight., (Copyright ©, International Association for Food Protection.)

Background: The effect of nanoemulsions prepared with grape seed and cinnamon essential oils on the shelf-life of flathead mullet (Mugil cephalus) fillets was evaluated by determining physicochemical (pH, free fatty acids, peroxide value, total volatile base nitrogen (TVB-N), and thiobarbituric acid reactive substances (TBARs)), sensory and microbiological (mesophilic aerobic bacteria, total psychrophilic bacteria, and Enterobacteriaceae counts) properties during 14 day storage at 2 °C., Results: The nanoemulsions showed good stability and low average droplet size. The results indicated that nanoemulsion treatments significantly prolonged the shelf-life of the fillets. Treatment inhibited increases in pH and TVB-N, and retarded lipid oxidation and hydrolysis. Sensory assessment revealed that treatment induced shelf-life extension from 10 to 14 days, compared with controls. Microbiological analyses showed nanoemulsion treatment caused shelf-life extension from 10 to 12 days with reduction of microbiological contamination by up to 1 log cfu g -1 in mesophilic and 1.5 log cfu g -1 in psychrotrophic bacteria., Conclusion: Considering the results, grape seed and cinnamon essential oil nanoemulsions could be considered as novel antimicrobial and antioxidant materials for shelf-life extension of flathead mullet fillets during cold storage. © 2021 Society of Chemical Industry., (© 2021 Society of Chemical Industry.)

This study was aimed to identify and optimize the culture conditions for gamma-aminobutyric acid (GABA) production by a lactic acid bacterium strain isolated from mam nem , a fermented fish sauce. Among the six isolates obtained from mam nem , the MN12 had the most potent GABA-producing capability. The strain was then identified to be Pedioccocus pentosaceus by employing MALDI-TOF-MS and phenylalanyl-tRNA synthase sequencing methods. The initial cell density of 5.10 6 CFU/mL, monosodium glutamate concentration of 60 mM, initial pH of 7, temperature of 45°C and cultivation time of 72 h were found to be the optimal culture conditions for highest production of GABA, reaching 27.9 ± 0.42 mM, by this strain. The cultivation conditions for GABA production by P. pentosaceus MN12 have been successfully optimized, providing a foundation for the development of fermented foods enriched with GABA.

Traditional high-salt fermented Suanyu is an ethnic fermented fish product in southwest China. Lactic acid bacteria (LAB) are the most appropriate strains because of their technological properties during ripening fermentation. The diversity of LAB in high-salt fermented Chinese Suanyu was examined through high-throughput sequencing (HTS), and the most suitable LAB strain was acquired through strain isolation and characterization, surimi simulation fermentation system, and principal component analysis (PCA). The processing adaptability of the strain was examined via Suanyu fermentation. Results showed that Lactobacillus, Tetragenococcus, and Weissella were the dominant bacteria in Suanyu, and their contributions were 53.99%, 35.60%, and 4.10%, respectively. The most suitable strain (Lactobacillus plantarum B7) rapidly produced acid, exhibited a strong antibacterial activity, showed salt tolerance, and had no amino acid decarboxylase activity. pH decreased to about 3.6. Eventually, the ability to tolerate 20% salt was observed, and the activity of amino acid decarboxylase was negative. Fermented Suanyu with B7 rapidly produced acid (11.7% d -1 ). The non-protein nitrogen (NPN) and total free amino acid (FAA) contents of fermented Suanyu were higher and its total volatile base nitrogen (TVB-N), thiobarbituric acid (TBARS), and biogenic amines (BAs) levels were lower than those of naturally fermented Suanyu. Therefore, B7 is a potential microbial starter for Suanyu industrial production., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

This study investigated the effect of antioxidants on lipid stability of mackerel (Scomber japonicus) fish balls. Oat phenolic acid compounds (OPC) and ascorbate palmitoyl (AP) were used to prolong the shelf life of steamed mackerel fish balls. Fish balls were stored at 4°C for 21 days, and the total bacterial count, hardness, whiteness, water holding capacity (WHC), pH, total volatile basic nitrogen (TVB-N), and thiobarbituric acid reactive substances (TBARS) value were monitored regularly. The results indicated that OPC+AP composite as a biological preservative could significantly inhibit the increase of the total bacterial count. Meanwhile, OPC and AP could maintain better hardness, whiteness, and WHC of fish balls during refrigerated storage. Furthermore, OPC and AP slowed down the increase of TVB-N and TBARS values. The results showed that OPC+AP had a synergistic effect on the storage time and could prolong the shelf life within the storage time. Adding OPC and AP was a promising strategy to improve the quality and shelf life of fish balls. PRACTICAL APPLICATION: The research provided a new application of OPC and AP for improving fish balls quality and shelf life during cold storage (4°C). OPC is a natural plant secondary metabolite from oat which exhibits excellent anti-oxidation. The research showed that OPC and AP combined with synergistic effect as biological preservatives can effectively inhibit the total bacterial count and reduce TBARS and TVB-N value of fish balls during the shelf life and maintain the hardness, which improved the quality and shelf life of fish balls., (© 2021 Institute of Food Technologists®.)

Fish and fish products are considered a fundamental part of the human diet due to their high nutritional value. Food-borne diseases are considered a major public health challenge worldwide due to their incidence, associated mortality, and negative economic repercussions. Food safety is the guarantee that foods will not cause harm to the health of those who consume them, and it is a fundamental property of food quality. Food safety can be at risk of being lost at any stage of the food chain if the food is contaminated by pathogenic microorganisms. Many diverse bacteria are present in the environment and as part of the microbiota of food that can be transmitted to humans during the handling and consumption of food. Plesiomonas shigelloides has been mainly associated with outbreaks of gastrointestinal diseases due to the consumption of fish. This bacterium inhabits the environment and aquatic animals and is associated with the microbiota of fish such as tilapia, a fish of importance in fishing, aquaculture, commercialization, and consumption worldwide. The purpose of this document is to provide, through a bibliographic review of databases (Scopus, Web of Science, and Google Scholar, among others), a general informative perspective on food-borne diseases and, in particular, the consumption of fish and tilapia. Diseases derived from contamination by Plesiomonas shigelloides are included, and control and prevention actions and sanitary regulations for fishery products established in several countries around the world are discussed to promote the safety of foods of aquatic origin intended for human consumption and to protect public health., Competing Interests: The authors declare that they have no conflicts of interest in the development and publication of this document., (Copyright © 2021 Alejandro De Jesús Cortés-Sánchez et al.)

Antimicrobial resistance (AMR) is a growing threat to human and animal health. However, in aquatic animals-the fastest growing food animal sector globally-AMR trends are seldom documented, particularly in Asia, which contributes two-thirds of global food fish production. Here, we present a systematic review and meta-analysis of 749 point prevalence surveys reporting antibiotic-resistant bacteria from aquatic food animals in Asia, extracted from 343 articles published in 2000-2019. We find concerning levels of resistance to medically important antimicrobials in foodborne pathogens. In aquaculture, the percentage of antimicrobial compounds per survey with resistance exceeding 50% (P50) plateaued at 33% [95% confidence interval (CI) 28 to 37%] between 2000 and 2018. In fisheries, P50 decreased from 52% [95% CI 39 to 65%] to 22% [95% CI 14 to 30%]. We map AMR at 10-kilometer resolution, finding resistance hotspots along Asia's major river systems and coastal waters of China and India. Regions benefitting most from future surveillance efforts are eastern China and India. Scaling up surveillance to strengthen epidemiological evidence on AMR and inform aquaculture and fisheries interventions is needed to mitigate the impact of AMR globally., (© 2021. The Author(s).)

This study aimed to achieve deeper insights into the microbiota composition and dynamic succession of the dry-cured black carp during storage using a high-throughput sequencing technique (HTS). The effect of lipid oxidation on microorganisms was also evaluated. Over 651 bacterial genera belonging to 37 phyla were identified. Firmicutes, Proteobacteria and Actinobacteria were the main bacterial phylum, some are highly associated with meat spoilage. Staphylococcus, Macrococcus and Acinetobacter were the most three microbial genera throughout the entire storage period (30 days). Between two different storage temperature, refrigeration at 4 °C could facilitate maintaining the microbial diversity, while 25 °C storage led to the formation of dominant microflora and the reduction of community diversity. Canonical correspondence analysis (CCA) showed that acid value (AV), malondialdehyde (MDA) and 4-hydroxy-2-hexenal (HHE) contents were three key environmental factors (oxidation products) affecting the profile of the microbiota. Staphylococcus presented a positive correlation with HHE content, while Macrococcus and Acinetobacter were negatively correlated with HHE content. These results could expand our knowledge on the effect of lipid oxidation on change of microbial distribution, it could also present an guideline to develop advanced storage methods for the vacuum packed dry-cured fish products., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Phenotypic and genotypic resistance to benzalkonium chloride (BC), cadmium and arsenic was tested (by susceptibility assays and molecular methods) in 287 Listeria monocytogenes strains isolated from fish and fish products, and food-producing factories in Poland. Overall, 40% of the isolates were resistant to BC, 56% to cadmium and 41% to arsenic (57% displayed resistance to more than one of the tested compounds). Among BC-resistant isolates, the most commonly detected resistance determinant was the qacH gene (83%). Three distinct types of cadA gene determining resistance to cadmium were detected, with the cadA1 variant predominant (88%), while most arsenic-resistant isolates (86%) harbored the arsA gene associated with a Tn554-like transposon (one strain harbored two copies of arsA in different arsenic resistance cassettes). 53% of all tested isolates contained plasmids (from 4 kb to > 90 kb in size), which were classified into 11 groups (p1-p11) based on their restriction patterns. Interestingly, 12 isolates harbored the small mobilizable pLMST6-like plasmid pLIS3 encoding multidrug efflux pump EmrC. Clustering analysis of PFGE patterns revealed that these isolates represent several diverse bacterial populations, which strongly suggests mobility of the pLMST6-like plasmids among L. monocytogenes strains and their role in dissemination of BC resistance., (Copyright © 2021. Published by Elsevier Ltd.)

Listeria monocytogenes is a potentially fatal foodborne pathogen that can be found in various ready-to-eat (RTE) products. It tolerates adverse conditions such as high salt concentrations and refrigerated storage, thus, the elimination of the pathogen in food processing often relies on heat processing. The objective of this study was to create a model to predict the effect of salt on heat tolerance of L. monocytogenes in meat and seafood products during heat treatments conducted at 57 to 65 °C to reduce numbers by ≥3 log 10 cycles. Salt concentrations, up to 6% in the water phase (WPS%), were applied to cover a variety of lightly salted RTE meat and seafood products. The experimental work involved samples of ground pork tenderloin, ground chicken breast fillet and skinned, ground salmon fillet adjusted to different WPS% i.e., 3.6 and 5.2 WPS% for pork samples, 2.0, 3.0, 3.5 and 6.0 WPS% for chicken samples and 3.0 and 6.0 WPS% for salmon samples. All samples were inoculated with late-stationary phase L. monocytogenes cultures. For pork samples, a two-strain mixture of a pork isolate (MS22254) and an environmental isolate (MS22246) was applied. For chicken and salmon samples, a seafood isolate (MS22258) and isolate MS22246 was applied as single cultures. Samples were vacuum-packed in sterile bags, immerged in water bath, and held at constant temperatures of 57, 60 and 65 °C for pork samples and 58, 61 and 62.5 °C for chicken and salmon samples. For survivor curves, where at least 3 log 10 -reduction were obtained, heat tolerance was expressed as decimal reduction times, D-values. D-values were observed to increase with increasing WPS%. The effect of salt on heat tolerance of L. monocytogenes was defined as the relative increase (RI-value) in D-value obtained when salt had been added to the food. The effect of WPS% on RI-values was independent of heating temperatures, foods and strains. For secondary modelling, RI-values were transformed using the natural logarithm, ln(RI) and fitted to a linear model as a function of WPS%. Model validation, with 56 independent values collected from the scientific literature, resulted in bias and accuracy factors of 0.89 and 1.26, respectively, suggesting acceptable performance with tendency to slightly under-predict. The developed predictive model can be used to guide the design of heat processes for manufacturers of lightly preserved and mildly processed meat and seafood products requiring more than 3 log 10 reduction of L. monocytogenes to ensure safety., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Psychrophiles have evolved to adapt to freezing environments, and cold-adapted enzymes from these organisms can maintain high catalytic activity at low temperature. The use of cold-adapted enzymes has great potential for the revolution of food and molecular biology industries., Results: In this study, four different strains producing protease were isolated from traditional fermented shrimp paste, one of which, named Planococcus maritimus XJ11 by 16S rRNA nucleotide sequence analysis, exhibited the largest protein hydrolysis clear zone surrounding the colonies. Meanwhile, the strain P. maritimus XJ11 was selected for further investigation because of its great adaptation to low temperature, low salinity and alkaline environment. The enzyme activity assay of P. maritimus XJ11 indicated that the optimum conditions for catalytic activity were pH 10.0 and 40 °C. Moreover, the enzyme also showed an increasing activity with temperatures from 10 to 40 °C and retained more than 67% activity of the maximum over a broad range of salinity (50-150 g L -1 ). Genome sequencing analysis revealed that strain XJ11 possessed one circular chromosome of 3 282 604 bp and one circular plasmid of 67 339 bp, with a total number of 3293 open reading frames (ORFs). Besides, 21 genes encoding protease, including three serine proteases, were identified through the NR database., Conclusion: Cold-adapted bacterium P. maritimus XJ11 was capable of producing alkaline proteases with high catalytic efficiency at low or moderate temperatures. Furthermore, the favorable psychrophilic and enzymatic characters of strain P. maritimus XJ11 seem to have a promising potential for industrial application. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

Amplicon sequencing approaches have been widely used in food bacterial ecology. However, choices regarding the methodology can bias results. In this study, bacterial communities associated with cold-smoked salmon products and their processing plant surfaces were monitored via sequencing of the V3-V4 region of the 16S rRNA gene. The impact of DNA extraction protocols, sampling methods (swabbing or sponging) and surface materials on bacterial communities were investigated. α and β diversity analyses revealed that DNA extraction methods mainly influence the observed cold-smoked salmon microbiota composition. Moreover, different DNA extraction methods revealed significant differences in observed community richness and evenness. β-Proteobacteria: Photobacterium, Serratia and Firmicutes: Brochothrix, Carnobacterium and Staphylococcus were identified as the dominant genera. Surface microbiota richness, diversity and composition were mainly affected by cleaning and disinfection procedures but not by DNA extraction methods. Surface community richness and evenness appeared higher when sampled by sponging compared to swabbing. β-diversity analyses highlighted that surface topology, cleaning and disinfection and sampling devices seemed to affect the bacterial community composition. The dominant surface bacteria identified were mainly Flavobacteriaceae, β-Proteobacteria and γ-Proteobacteria described as fish spoilers such as Acinetobacter, Pseudomonas and Shewanella. DNA extraction and sampling methods can have an impact on sequencing results and the ecological analysis of bacterial community structures. This study confirmed the importance of methodology standardization and the need for analytical validation before 16S rDNA metabarcoding surveys., (Copyright © 2020. Published by Elsevier Ltd.)

In this study, the bioprotective potential of Lactobacillus sakei CTC494 against Listeria monocytogenes CTC1034 was evaluated on vacuum packaged hot-smoked sea bream at 5 °C and dynamic temperatures ranging from 3 to 12 °C. The capacity of three microbial competition interaction models to describe the inhibitory effect of L. sakei CTC494 on L. monocytogenes was assessed based on the Jameson effect and Lotka-Volterra approaches. A sensory analysis was performed to evaluate the spoiling capacity of L. sakei CTC494 on the smoked fish product at 5 °C. Based on the sensory results, the bioprotection strategy against the pathogen was established by inoculating the product at a 1:2 ratio (pathogen:bioprotector, log CFU/g). The kinetic growth parameters of both microorganisms were estimated in mono-culture at constant storage (5 °C). In addition, the inhibition function parameters of the tested interaction models were estimated in co-culture at constant and dynamic temperature storage using as input the mono-culture kinetic parameters. The growth potential (δ log) of L. monocytogenes, in mono-culture, was 3.5 log on smoked sea bream during the experimental period (20 days). In co-culture, L. sakei CTC494 significantly reduced the capability of L. monocytogenes to grow, although its effectiveness was temperature dependent. The LAB strain limited the growth of the pathogen under storage at 5 °C (<1 log increase) and at dynamic profile 2 (<2 log increase). Besides, under storage at dynamic profile 1, the growth of L. monocytogenes was inhibited (<0.5 log increase). These results confirmed the efficacy of L. sakei CTC494 for controlling the pathogen growth on the studied fish product. The Lotka-Volterra competition model showed slightly better fit to the observed L. monocytogenes growth response than the Jameson-based models according to the statistical performance. The proposed modelling approach could support the assessment and establishment of bioprotective culture-based strategies aimed at reducing the risk of listeriosis linked to the consumption of RTE hot-smoked sea bream., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Bacteria release membrane vesicles into the extracellular environment but which activity is unclear. We investigated the applications of extracellular vesicles (EVs) isolated from probiotic Lactobacillus plantarum to protect tuna fish against spoilage and quality loss in this study. A significant difference was found in EVs size obtained from L. plantarum after 8, 24, and 48 hr incubation. The L. plantarum-derived EVs were collected and used to confirm the anti-bacterial activity versus Shewanella putrefaciens. Finally, the tuna fish was stored at 4 °C for 5 days after coating with EVs or sodium erythorbate, and the quality indexes were assayed. Results indicated that EVs markedly inhibited oxidation reaction, total volatile base nitrogen (TVBN), peroxide value (PV), malondialdehyde (MDA), and bacteria levels. These results finding out that EVs from L. plantarum may have potential for application in food storage technology. Overall, we indicated this new material may be developed as an anti-bacterial agent for prolonging the shelf life of tuna fish., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

DNA-sequencing was performed on the V3-V4 regions of 16S rRNA genes to investigate the microbial diversity of five samples of fermented freshwater fish (pla-ra) from three provinces in northeastern Thailand. The samples had salt concentrations ranging from 7 to 10%, pH values from 4.83 to 7.15, and D-/L-lactic acid concentrations of 90 to 450 mg/l. A total of 598 operational taxonomic units were annotated at various taxonomic ranks based on the SILVA Database. The lactic-acid and halophilic genera Tetragenococcus, Halanaerobium and Lactobacillus were among the dominant taxa of bacteria. The top 20 non-redundant taxa were considered in more detail. In two pla-ra samples, Tetragenococcus muriaticus was commonly identified. Halanaerobium fermentans was the most abundant species in a third sample and co-dominant in another sample. Lactobacillus rennini was dominant in the pla-ra sample from Roi Et Province. Additionally, other beneficial bacteria were detected including Staphylococcus nepalensis, Lactobacillus sakei, Lactobacillus pentosus, Weissella confusa, and Bifidobacterium bifidum. Differences between samples may be due to use of different raw materials, salt concentrations, recipes, processes and fermentation periods. The microbial communities in pla-ra provide a better understanding of the production outcomes of traditional products. Further optimization of the fermentation process, for example by using dominant bacterial taxa in starter cultures, may improve processes of food fermentation, food quality and flavor control, providing useful guidelines for industrial applications., Competing Interests: The authors have declared that no competing interests exist.

Fresh Atlantic salmon (Salmo salar) represents a healthy, nutritious food with global distribution and increasing consumption and economic value. Contaminating Listeria monocytogenes in fresh salmon represents a health hazard to consumers, is linked to extensive product recalls and is a major challenge for salmon processors. Verdad N6, a commercially available buffered vinegar, was evaluated as a treatment for raw salmon fillets either alone or in combination with the antimicrobial peptide nisin, with regard to anti-listerial effects under processing and storage, and influence on sensory quality and background microbiota. Salmon fillets were surface contaminated with L. monocytogenes and immersed in solutions of Verdad N6 or treated with nisin or a combination of these two treatments. Levels of L. monocytogenes were determined during vacuum-pack refrigerated storage. The use of Verdad N6 resulted in increased lag times and substantially reduced growth of L. monocytogenes. The inhibitory effects were dependent on Verdad N6 levels, immersion time, and storage time and temperature. A 5 s immersion in 10% Verdad N6 solution at 4 °C reduced growth of L. monocytogenes from log 2.8 to log 1 after 12 days of storage. Nisin (0.2-1 ppm) had listericidal effects up to 1 log but did not inhibit regrowth when used alone. Appropriate combinations of Verdad N6 and nisin led to L. monocytogenes levels no higher after 12 days of storage than the initial levels. The inhibitory effects were markedly lower at 7 °C than at 4 °C. Salmon with Verdad N6 showed reduced levels of total counts during storage indicating a longer shelf-life, and a shift in the dominating bacteria with reduced and increased relative levels of Enterobacteriaceae and lactic acid bacteria, respectively. Sensory analyses of raw and cooked Verdad N6 treated a non-treated salmon resulted in small differences. In summary, Verdad N6 is an option for production of high-quality raw salmon with increased shelf-life and enhanced food safety through its Listeria inhibiting effects. The application of Verdad N6 in combination with nisin treatment can further reduce the listeria-risks of these products., (Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.)

Listeria monocytogenes , an important foodborne pathogen, may be present in different kinds of food and in food processing environments where it can persist for a long time. In this study, 28 L. monocytogenes isolates from fish and fish manufactures were characterized by whole genome sequencing (WGS). Core genome multilocus sequence typing (cgMLST) analysis was applied to compare the present isolates with publicly available genomes of L. monocytogenes strains recovered worldwide from food and from humans with listeriosis. All but one (96.4%) of the examined isolates belonged to molecular serogroup IIa, and one isolate (3.6%) was classified to serogroup IVb. The isolates of group IIa were mainly of MLST sequence types ST121 (13 strains) and ST8 (four strains) whereas the isolate of serogroup IVb was classified to ST1. Strains of serogroup IIa were further subtyped into eight different sublineages with the most numerous being SL121 (13; 48.1% strains) which belonged to six cgMLST types. The majority of strains, irrespective of the genotypic subtype, had the same antimicrobial resistance profile. The cluster analysis identified several molecular clones typical for L. monocytogenes isolated from similar sources in other countries; however, novel molecular cgMLST types not present in the Listeria database were also identified.

Aroma defects limit the application of fish protein hydrolysates as flavourings. This study aimed to develop a flavour concentrate from fermented tilapia fish head hydrolysate bymaximising the Maillard reaction production of meaty and roasted aroma associated compounds. We studied the optimal conditions of the Maillard reaction of xylose with cysteine to form meat-like odorants using response surface methodology. A 3-factored and 3-leveled Box Behnken design was employed, where the independent variables were cysteine concentration (A, w/v, %), heating temperature (B, °C) and heating time (C, min). 2-Methyl-3-furanthiol and 2-furfurylthiol were used as response factors. The optimal conditions were obtained as follows: A, 0.80%; B, 183.80 °C; C, 89.34 min. Compared with the controls, Maillard reaction products enriched the meaty and roasted aroma associated compounds in the treated hydrolysate. In conclusion, the treated tilapia fish head hydrolysate may be used as a base in development of new fish-based flavourings., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Staphylococci from Sheedal of Northeast India was isolated, identified and characterized. All the isolated staphylococci were found to be coagulase negative. Based on the rpoB gene sequences followed by analysis using NCBI-BLAST software, seven species of Staphylococcus namely, S. piscifermentans, S. condimenti, S. arlettae, S. sciuri, S. warneri, S. nepalensis and S. hominis were recognized. Phylogenetic analyses revealed three major cluster groups. All the seven Staphylococcus showed their NaCl tolerance from 2 to 8%. No species was able to grow at 55°C. Except S. arlettae and S. sciuri, all the isolated staphylococcal species exhibited growth at pH 4-8. No isolated species was able to ferment mannitol, sucrose and arabinose. All the species exhibited moderate to maximum proteolytic and lipolytic activities. All the seven species were found to be sensitive to the antibiotics, namely, erythromycin, norfloxacin, ampicillin, streptomycin and vancomycin, whereas all were resistant to co-trimoxazole. Only S. piscifermentans was found antagonist to Salmonella enterica, Escherichia coli and Bacillus subtilis, although the clear zone was minimal. All the staphylococcal species except S. arlettae and S. sciuri exhibited hydrophobicity ranging from 25 to 66%. The observed characteristics of isolated Staphylococci from Sheedal revealed their role in fish fermentation., (© 2020 The Society for Applied Microbiology.)

Naturally occurring fungi have been used in the traditional production of dried bonito, Katsuobushi, in Japan. In this study, we analyzed the fungal population present during Katsuobushi production. Amplicon sequence analysis of ITS1 indicated that Aspergillus spp. are predominant throughout the production process. In addition, culture-dependent analyzes identified three species Aspergillus chevalieri, Aspergillus montevidensis, and Aspergillus sydowii, based on sequencing of benA, caM, and rpb2 genes. A. chevalieri isolates were classified into teleomorphic and anamorphic strains based on morphological analysis. A. chevarieri was the dominant species throughout the production process, whereas A. montevidensis increased and A. sydowii decreased in abundance during Katsuobushi production. Our study will enhance the understanding of fungal species involved in traditional Katsuobushi production.

A 16S rRNA next-generation sequencing technique was applied to investigate the microbial diversity and liquid chromatography-tandem mass spectrometry was used to identify glutamyl peptide profiles of 10 Thai fermented freshwater fish (Pla-ra) samples. A total of 12 genera of bacteria were able to be detected, with Tetragenococcus spp., Staphylococcus spp., and Lactobacillus spp. dominating. Of the 18 glutamyl peptides analyzed, 17 were found, even though the amounts detected were lower than the taste threshold. Despite this, an increase in mouthfulness sensation, reflecting kokumi activity, was clearly identified in most of the samples, which might be because of a synergistic effect of different sub-threshold compounds present in the samples. In principle component analysis, the relationship between microorganisms and glutamyl peptide generation was observed, especially between Tetragenococcus spp. and Lentibacillus spp. and the generation of γ-Glu-Val-Gly. Correlations between microbial diversity and the generation of taste enhancers were identified in this study.

Background: An innovative pilot-plant packaging was developed and evaluated for applying oregano essential oil (OEO) vapours in conditions of high vacuum for exploring the antimicrobial effect of essential oil vapours applied immediately before packaging of fish fillets. Farmed sea bream (Sparus aurata) fresh fillets have been used as a model for validating this new technology. These fillets, as a refrigerated product under modified atmosphere packaging (MAP), have a relatively short shelf life (12-14 days) mainly due to the fast microbial growth. The effects of conventional OEO dippings [pretreatment dipping (0.1% of OEO) of whole fish (T1) and filleted sea bream (T2)] were compared with the OEO application in vapour phase (67 μL L -1 ) under vacuum (5-10 hPa) immediately before MAP fillet packaging (T3)., Results: T3/T2 samples showed the lowest microbial growth after 28 days at 4 °C, with loads up to 1/2.6 log units for Enterobacteria/lactic acid bacteria compared to untreated samples. The initial trimethylamine nitrogen (TMA-N) content (2.6 mg kg -1 ) increased in T1 and T2/T3 samples by 9.6 and 6/7 units, respectively, after 28 days. Quality Index Method (QIM) better reflected the fish fillets shelf life than texture and colour measurements. The shelf life of T3/T2 samples was established in at least 28 days (4 °C), while the QIM threshold (6) was exceeded after 7/21 days in untreated/T1 fillets., Conclusion: The fish shelf life was extended with vapour OEO treatment using this new technology, similarly to OEO dipping treatment, according to QIM, corroborated by the microbial quality and TMA-N contents. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)

A soup stock made from katsuobushi is an important element of, and the basic seasoning responsible for the taste of, traditional Japanese cuisine. Fermented and ripened katsuobushi, called karebushi, is manufactured via a repeated molding process on the katsuobushi surface. Our aim was to characterize the surface Aspergillus community and their enzymes involved in the fermentation and ripening. Five dominant Aspergillus species isolated from the karebushi surface were identified-A. amstelodami, A. chevalieri, A. pseudoglaucus, A. ruber, and A. sydowii. Analyses were performed on final molding stage-samples from different manufacturers, and 1st to 4th molding stage-samples from the same manufacturer. The composition ratios of the five Aspergillus spp. varied according to the manufacturer of the karebushi. A. amstelodami and A. chevalieri tended to be detected as dominant species when the water content of the karebushi fillet was >15% and the fat content was >3.5%, respectively. In samples from a given manufacturer, the dominant species in the final molding stage tended to be A. chevalieri and A. pseudoglaucus. Mixed molds were cultured by solid-state fermentation using katsuobushi powder medium at two different water activity (a w ) levels. Crude extracts from each culture showed lipase, aminopeptidase, carboxypeptidase, and protease activities. Notably, the crude extracts cultivated at 0.85 a w showed higher protease activity toward hemoglobin and lipase activity toward p-nitrophenyl palmitate than those at 0.95 a w . These hydrolytic enzymes are probably involved in decolorization of katsuobushi and lipid degradation during the long fermentative and ripening period. In addition, mixed cultures could transform 2,6-dimethoxyphenol into 1,2,3-trimethoxybenzene, previously reported as an attractive and mild flavor component. Our results may help promote the use of desirable Aspergillus spp. as starter cultures for manufacturers to stabilize and improve the quality of fermented and ripened karebushi., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This project aimed to evaluate the effects of gallic acid (GA) and protocatechuic acid (PA) grafted onto chitosan (CS) on the improved quality of sea bass (Lateolabrax japonicus) during refrigerated storage. The incorporation of GA and PA onto CS (CS-g-GA and CS-g-PA) were achieved by the carbodiimide-mediated grafting procedure. Samples were treated with different solutions (deionized water [CK], 1% CS [m/v], 1% CS-g-GA [m/v], and 1% CS-g-PA [m/v]) for 10 min, which were then stored at 4 °C. Microbiological quality, including total viable counts (TVC), psychrophilic bacterial counts (PBC), Pseudomonas bacterial counts, and H 2 S-producing bacterial counts were measured. Physicochemical parameters, including pH, total volatile basic nitrogen (TVB-N), thiobarbituric acid (TBA) value, water holding capacity (WHC), and K value, were measured. The changes in protein characteristics, including sulfhydryl groups (SH), Ca 2+ -ATPase activity, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and tertiary structure of protein were analyzed periodically, along with texture profile analysis (TPA). The results demonstrated that the CS copolymers treatment exhibited better preservation effects. The CS-g-GA and CS-g-PA treatments could significantly inhibit the growth of microorganisms and retard the increase of pH, TVB-N, TBA, WHC, and K-value during refrigerated storage compared with the CK and CS groups. Additionally, the CS-g-GA and CS-g-PA treatments could delay the protein oxidation by keeping a higher SH level and Ca 2+ -ATPase activity. The CS copolymers treatment could also extend the shelf life for another 6 days compared with that of CK. As a result, CS copolymers can be employed in a promising method for the preservation of sea bass. PRACTICAL APPLICATION: The incorporation of gallic acid and protocatechuic acid onto chitosan (CS-g-GA and CS-g-PA) showed superior antioxidant and antimicrobial activities when applied on sea bass. The CS-g-GA and CS-g-PA coatings could maintain the quality and freshness of refrigerated sea bass. Additionally, this research could provide a theoretical basis for the application of graft copolymers on the preservation of aquatic products., (© 2020 Institute of Food Technologists®.)

Acid adaptation enhances survival of foodborne pathogens under lethal acid conditions that prevail in several food-related ecosystems. In the present study, the role of undissociated acetic acid in inducing acid resistance of Salmonella Enteritidis Phage Type 4 both in laboratory media and in an acid food matrix was investigated. Several combinations of acetic acid (0, 15, 25, 35 and 45 mM) and pH values (4.0, 4.5, 5.0, 5.5, 6.0) were screened for their ability to activate acid resistance mechanisms of pathogen exposed to pH 2.5 (screening assay). Increased survival was observed when increasing undissociated acetic acid within a range of sublethal concentrations (1.9-5.4 mM), but only at pH 5.5 and 6.0. No effect was observed at lower pH values, regardless of the undissociated acetic acid levels. Three combinations (15mM/pH5.0, 35mM/pH5.5, 45mM/pH6.0) were selected and further used for adaptation prior to inoculation in commercial tarama (fish roe) salad, i.e., an acid spread (pH 4.35 ± 0.02), stored at 5°C. Surprisingly and contrary to the results of the screening assay, none of the acid adaptation treatments enhanced survival of Salmonella Enteritidis in the food matrix, as compared to non-adapted cells (control). Further examination of the food pH value, acidulant and storage (challenge) temperature on the responses of the pathogen adapted to 15mM/pH5.0, 35mM/pH5.5 and 45mM/pH6.0 was performed in culture media. Cells adapted to 35mM/pH5.5 were unable to induce acid resistance when exposed to pH 4.35 (tarama salad pH value) at 37°C and 5°C, whereas incubation under refrigeration (5°C) at pH 4.35 sensitized 45mM/pH6.0 adapted cells against the subsequent acid and cold stress. In conclusion, pre-exposure to undissociated acetic acid affected the adaptive responses of Salmonella Enteritidis Phage Type 4 in a concentration- and pH-dependent manner, with regard to conditions prevailing during acid challenge., Competing Interests: The authors have declared that no competing interests exist.

This study investigated the effect of phytic acid and lysozyme on the microbial composition and quality of grass carp (Ctenopharyngodon idellus) fillets stored at 4 °C. The control, 0.5 mg/mL lysozyme-treated fillets (T1), 0.5 mg/mL phytic acid-treated fillets (T2) and 0.25 mg/mL lysozyme + 0.25 mg/mL phytic acid-treated fillets (T3) were evaluated based on sensory assessment, biogenic amines, ATP-related compounds, total volatile basic nitrogen (TVB-N), and total viable counts (TVC). Changes in microbial composition were analyzed using high-throughput sequencing. Results showed that phytic acid and lysozyme treatment delayed the decrease in sensory scores, reduced the rate of degradation of IMP to Hx, inhibited the growth of microorganisms, and attenuated the increase in TVB-N and putrescine. Phytic acid exhibited better preservation effects than lysozyme and their combination was more effective than using either alone. High-throughput sequencing showed that Acinetobacter and Kocuria were the predominant bacteria in fresh grass carp, but Pseudomonas rose rapidly with storage time; Pseudomonas, Shewanella, and Aeromonas constituted the main spoilage bacteria of grass carp fillets. Lysozyme treatment significantly reduced the proportion of Shewanella and Acinetobacter, and phytic acid and the combination of phytic acid and lysozyme significantly reduced the proportion of Pseudomonas in spoiled grass carp fillets., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

The microbiological quality of thirty ready-to-eat (RTE) keropok lekor (a sausage shape Malaysian fish product) was evaluated in comparison to microbiological guidelines for ready to eat foods. The two E. coli isolates were subjected to DNA sequencing, identified and tested for their resistance towards fifteen different antibiotics. The survival and growth of the isolated E. coli strains inoculated in keropok lekor at atmospheric air and vacuum packaging were also evaluated. Results revealed that four samples (13.33%) contained Enterobacteriaceae counts that exceeded the recommended allowable counts of 4.0 log 10 CFU/g. Unsatisfactory level of coliforms (< 1.7 log 10 CFU/g) was also observed in ten of the samples; two of which contained E. coli (2.1 ± 0.17 and 3.7 ± 0.02 log 10 CFU/g), suggesting of poor hygiene and sanitation practices. While the 'Possible E10' E. coli strain was observably resistant towards Nalidixic acid (30µg) alone, B10 E. coli isolate was worryingly resistant towards Ampicillin (10µg), Ceftazidime (30µg), Ciprofloxacin (5µg), Ceftriaxone (30µg), Nalidixic acid (30µg) and Tetracycline (30µg). This study also revealed that the growth and survival of the 'Possible E10' and B10 E. coli strains were not significantly affected by vacuum packaging when stored at both 4°C and 28°C. Therefore, intervention programmes to alert and educate smallmedium enterprisers (SMEs) of keropok lekor producers on food safety as well as potential health risks that can be associated due to inappropriate handling procedures of such product, merits consideration.

The variation in microbial composition over time was assessed in biofilms formed in situ on selected non-food and food contact surfaces of meat and fish industries, previously identified as Listeria monocytogenes-positive foci. First, all samples were analysed for the detection and quantification of L. monocytogenes using ISO 11290-1 and ISO 11290-2 norms, respectively. Although the pathogen was initially detected in all samples, direct quantification was not possible. Psychrotrophic bacteria counts were among resident microbiota in meat industry samples (Mean max  = 6.14 log CFU/cm 2 ) compared to those form fish industry (Mean max  = 5.85 log CFU/cm 2 ). Visual analysis of the biofilms using epifluorescence microscopy revealed a trend to form microcolonies in which damaged/dead cells would act as anchoring structures. 16S rRNA gene metagenetic analysis demonstrated that, although Proteobacteria (71.37%) initially dominated the bacterial communities at one meat industry location, there was a dramatic shift in composition as the biofilms matured, where Actinobacteria (79.72%) became the major phylum present in later samples. This change was largely due to an increase of Nocardiaceae, Micrococcaceae and Microbacteriaceae. Nevertheless, for the other sampling location, the relative abundance of the dominating phylum (Firmicutes) remained consistent over the entire sampling period (Mean = 63.02%). In fish industry samples, Proteobacteria also initially dominated early on (90.69%) but subsequent sampling showed a higher diversity in which Bacteroidetes and Proteobacteria were the most abundant phyla accounting for the 48.04 and 37.98%, respectively by the last sampling period. Regardless of the location, the community profiles of the endpoint samples were similar to those reported previously. This demonstrated that in a given industrial setting there is a trend to establish a determinate biofilm structure due to the environmental factors and the constant incoming microbiota. This information could be used to improve the existing sanitisation protocols or for the design of novel strategies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The population, diversity and succession of microbial communities and chemical characteristics of dried crimson snapper (Lutianus erythropterus) during storage for 50 days were investigated. The population of bacteria, yeasts and filamentous fungi in the samples were enumerated by culture methods using appropriate agar media. The amplicons of the 16S rRNA of bacteria and the ITS region of fungi were sequenced and compared with gene libraries to obtain the identity and abundance of microorganisms in the community. Free amino acids and several other chemical characteristics were determined by HPLC and corresponding chemical reaction methods. Before storage, the average counts of bacteria, yeasts and filamentous fungi in the dried fish were 3.2, 2.5 and 2.2 log CFU/g, which increased to 4.6, 3.6 and 3.9 log CFU/g, respectively after storage. Major succession in the bacterial > fungal communities occurred during storage as evidenced by a significant decline in the number and diversity of microbial communities. Predominant bacterial genera were Phytobacteria, Vibrio, Acinetobacter and Macrococcus in the freshly dried fish, which were replaced by Bacillaceae, Halomonas, Lentibacillus, Alkalibacillus after storage. The fungal community of the freshly dried fish consisted of Penicillum > Yamadazyma, Malassezia, Candida and Eurotiales with Penicillum being the most dominant. Penicillium camemberti was the most abundant fungal species in the dried fish before storage with most dominant after storage and accounted for >96% of the abundance. The succession in the microbial community was accompanied by major changes in chemical characteristics with a significant decrease in fat, and an increase in total free amino acids and total volatile basic nitrogen (TVB-N)., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Fermentation is a traditional food preservation method and is widely used for improving food safety, shelf life, and organoleptic and nutritional attributes. Fermented fish are produced and consumed in different parts of the world and are an integral part of many food cultures. Furthermore, fermented fish are a source of interesting microbes and are an important industry in many countries. This review tries to update the types and manufacturing processes for fermented fish around the world. The emphasis is on this work related to fermented fish and their health benefits, as well as the contribution of microorganisms to their fermentation. A variety of different approaches have been used to determine and understand microbial composition and functionality. Moreover, some challenges and future research directions regarding fermented fish are also discussed in this review. Further research into fermented fish products is of crucial importance not only for the food industry but also for human health. However, extensive in vivo and toxicological studies are essential before the application of bioactive-rich fermented fish products for human health benefits.

Background and Objective: Shiga toxin-producing Escherichia coli O157:H7 is a troubled foodborne pathogen associated with contamination of meat, fish and poultry. The present work aimed to evaluate plant extracts as natural preservatives anti- Escherichia coli O157:H7 in meat and fish products., Materials and Methods: Antibacterial activity and minimum inhibitory concentrations (MICs) of seven herbal plants, clove, marjoram, sage, pomegranate peel, turmeric, Cassia fistula and black pepper and their different 6 mixes were examined against Escherichia coli O157:H7. Phytochemical qualitative analysis, phenolic compounds (HPLC), total phenolic, total flavonoid contents and antioxidant activities of individual extracts and their 6 mixes were evaluated. Combination Mix 5 extract was applied on meat and fish-fillet, then its antimicrobial effect against E. coli O157:H7 and sensory evaluation were assessed., Results: Five extracts exhibited good antibacterial activity against Escherichia coli O157:H7. The greatest inhibition zone was recorded by clove aqueous extract (25 mm). Mix 5 (clove, sage, pomegranate and Cassia fistula) showed the highest inhibition with MIC of 3.0 mg mL-1. This mix exhibited strong anti-bactericidal effect against E. coli O157:H7 in meat and fish-fillet products throughout 8 days of cold storage (4°C). The sensory evaluation revealed that Mix 5 was acceptable by panelists with concentration of 0.50% in beef burgers and 0.25% in fish-fillet., Conclusion: This study suggests that the use of herbal extracts provide antibacterial potentials against food pathogens in meat and fish products.

Low-temperature cooking, such as sous vide, has become a favored method for processing seafood. For this method to be applicable for retail products, combinations with other processing steps are needed to keep the products safe and durable while maintaining high quality. The present experiments were designed to investigate the influence of low-temperature treatment (40, 50, or 60 °C) in combination with various packaging technologies (modified atmosphere [MA] or soluble gas stabilization [SGS]) on both the microbial growth and the physiochemical quality. Salmon loins were either kept natural or inoculated with Listeria innocua prior to drying (16 to 18 hr) in either 100% CO 2 (SGS) or atmospheric air (MA packaging). All samples were sous vide treated, repackaged in MA, and stored at 4 °C for 24 days. The results showed shelf life to be significantly improved with the implementation of SGS, by prolonging the lag-phase and slowing the growth rate of both naturally occurring and inoculated bacteria. Variations in packaging technology did not significantly influence any of the tested quality parameters, including drip loss, surface color, and texture. Growing consumer demand for lightly processed seafood products makes Listeria spp. an increasing problem. The present experiment, however, has shown that it is possible to lower processing temperatures to as little as 40 or 50 °C and still obtain inhibition of Listeria, but with improved chemical quality compared to traditional processing., (© 2019 Institute of Food Technologists®.)

Background: Probiotic bacteria can provide health benefits when delivered in functional foods. This study involved isolation of lactic acid bacteria (LAB) from traditionally dried and salted anchovy fish and characterization of their survival in simulated gastrointestinal digestion. Promising strains were used to prepare fermented fish sausages which were then evaluated for cytotoxicity activity against two cancer cell-lines, antidiabetic activity as determined by α-amylase and α-glucosidase inhibition, and antioxidant and proteolytic activities in vitro, as compared to non-fermented control sausages., Results: Out of 85 LAB obtained, 13 isolates with high tolerance to simulated gastrointestinal digestion were obtained, which were identified as Enterococcus spp. Four E. faecium strains, one E. faecalis, and one E. durans were used separately to make fermented fish sausages. The α-amylase and α-glucosidase inhibition from fish sausages fermented by Enterococcus spp. ranged from 29.2 to 68.7% and 23.9 to 41.4%, respectively, during 21 days of storage. The cytotoxicity activities against Caco 2 and MCF-7 cells of fish sausages fermented with Enterococcus spp. ranged from 18.0 to 24% and 13.9 to 27.9%, respectively. Cytotoxicity activities correlated positively with proteolysis and antioxidant activities, α-amylase and α-glucosidase inhibition activities, but negatively with the pH in fermented fish sausages. Strains also exhibited antimicrobial activity against foodborne pathogens and presented no significant concerns with regards to antibiotic resistance or virulence gene content., Conclusions: Fish sausages fermented by potential probiotic isolates of Enterococcus spp. from dried fish had valuable health-promoting benefits compared with non-fermented control sausages.

We developed a technological accessory bacteriophage-based preparation and a method for phage-mediated bioprocessing for elimination of pathogenic microorganisms from the surface of fresh fish and for prolongation of the shelf-life of chilled hydrobionts. Specimens of rainbow trout (Salmo irideus) served as the objects of the study carried out at a fish-processing plant in the Republic of Karelia. The specimens were decontaminated by a bacteriophage cocktail containing six original virulent phage strains characterized by their pheno- and genotypical properties. A new method of biodecontamination (plunging the rainbow trout for 30 sec into a solution of bacteriophage cocktail (bacteriophage titers ≥10 8 PFU/ml) delayed bacterial degradation of hydrobionts by 3 days. The use of the new method for decontamination of food half-products - phage-mediated bioprocessing - promoted preservation of the initial ecological purity, nutritive value, and taste of the products and prolonged their shelf-life in comparison with the actual standards.

This study examined the prevalence and phenotypic and genotypic antibiotic resistance patterns of Staphylococcus aureus in sushi from 20 Danish outlets. Microbial quality of sushi products and food inspector ranking of outlets were assessed and results for thirteen of the outlets were compared with findings from a previous study in 2012. Inspector rankings were similar in the two studies. The mesophilic aerobic counts were slightly lower (p = 0.0296) in 2017 than in 2012 with average values of the 13 shops of 5.2 log CFU/g and 5.7 log CFU/g, respectively. In both studies E. coli was only found in the products from outlets that did not have consistently superior rankings. On the other hand prevalence and average counts of Staphylococcus spp. were slightly higher in 2017 (p = 0.0286) but no methicillin-resistant S. aureus (MRSA) were observed in the present study. Methicillin-sensitive S. aureus (MSSA) were, however, isolated from 18.7% of sushi products with an average count below 2 log CFU/g. Based on spa and multilocus sequence typing (MLST), isolates belonged to clonal complex CC7 (t2016), CC20 (t7836), CC45 (t065, t127, t362), CC88 (t1998) and CC398 (t164, t331, t1451). The Panton-Valentine leukocidin (PVL)-encoding gene lukF was detected only in isolates of the t065 spa-type whereas the scn gene from the ΦSa3 prophage was detected in 76.5% of the isolates, supporting that the majority of isolates were of likely human origin. Thirty-six isolates (70.6%) were resistant to at least one of the antibiotic compounds tested. Antibiotic resistance genes that confer resistance to β-lactams (blaZ) and macrolides (ermC) were detected in 33.3% and 9.8% of isolates, respectively. The tet(K) gene that encode tetracycline resistance was only found in a t7836 strain. Overall, this study indicates that S. aureus in sushi products in Denmark do not represent a major food safety hazard due to, firstly, the low temperature and limited time of storage of product may prevent significant growth and production of toxic levels of enterotoxin of this species. Secondly, the S. aureus isolates obtained did not include MRSA variants and none of them encoded PVL that constitute one of the virulence factors in pathogenesis. Several MSSA isolates contained however genes encoding antibiotic resistance, which emphasize the potential role of foods as vehicles for transmission of such variants., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Biogenic amines (BAs) are frequently present in traditionally fermented salted foods. In this study, a Tetragenococcus halophilus strain (MJ4) with no BA-producing ability was isolated from a fish (anchovy) sauce. Strain MJ4 did not produce BAs from supplied precursors and no BA-producing genes were identified in its genome. Bacterial community analysis showed that in non-inoculated saeu-jeot (shrimp sauce) fermentation, Tetragenococcus predominated after 82 days, while in strain MJ4-inoculated saeu-jeot, Tetragenococcus predominated during the entire fermentation. Strain MJ4 repressed the growth of T. muriaticus, a known BA producer, during fermentation, but metabolite analysis demonstrated that metabolite profiles, including amino acids, were similar regardless of MJ4 inoculation. The metabolite analysis also showed that strain MJ4 clearly repressed the formation of cadaverine during fermentation. This study suggests that the use of strain MJ4 as a starter culture in salted fish fermentation may be a good strategy for the reduction of BA formation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Lactic acid bacteria and Listeria monocytogenes are psychotropic organisms that can grow and compete in food such as lightly preserved fishery products. Predictive microbiology is nowadays one of the leading tools to assess the behavior of bacteria in food and to predict food spoilage. Mathematical models can be used to predict the growth, inactivation or growth probability of bacteria. Currently, the efforts in microbial modeling are oriented towards extrapolation of results beyond experiments in order to predict the growth of interacting microorganisms and develop new food preservation processes. In the present work, a model combining both heterogeneous population and quasi-chemical approaches to describe the different phases of the bacterial growth curve is presented. The model was applied to both monoculture and co-culture cases of lactic acid bacteria, Carnobacterium maltaromaticum H-17, and two Listeria monocytogenes strains in a raw fish extract. It is a highlight that our model includes novel inhibition reactions due to the accumulation of metabolites, and a general equation to take into account the effect of chemical compounds during the lag or physiological adaptation phase of the cells. Our results show that the proposed model can accurately describe the experimental data when the curve shape is a sigmoid, and when it presents a maximum. Besides, the parameters have biological interpretability since the model is mechanistically inspired., (© 2019 Institute of Food Technologists®.)