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The aim of the present work was to evaluate the effect of various hurdles such as aw and pH as well as the storage atmosphere on the microbiological and sensory changes of minimally processed (lightly brined or marinated with acetic or citric acid) European sea bass (Dicentrarchus labrax) fillets. The results indicated that the shelf-life of brined fillets stored aerobically was 6 d while that of vacuum and MAP-stored was 12 d, since a reduced growth rate of spoilage bacteria was recorded. The physicochemical characteristics such as aw and water phase salt (WPS) were not considerably changed, while the oxygen levels into the packages ensure the microbiological safety of the product. The fillets marinated with acetic acid exhibited a longer shelf-life at 30 and 40 d under aerobic and reduced oxygen conditions, respectively, while the products marinated with citric acid were at 25 and 35 d respectively. A low pH resulted in reduced or even limited microbial levels, especially for the fillets marinated with acetic acid; something that ensures microbiological safety as well. The low or limited microbial levels in conjugation with the sensory attributes indicated that spoilage may be due to other mechanisms such as autolysis rather than microbial activity. Overall, the present work highlights the potential for further research and development of minimally processed, microbiologically safe and stable with extended shelf-life value added seafood to satisfy the corresponding consumer demands.

Microbial spoilage is the main cause of quality deterioration in seafood. Several strains of psychotropic Pseudomonas have been found to dominate in such products, producing a plethora of volatile organic compounds (VOC). Herein, we investigated the growth of and VOC production by seven strains of Pseudomonas associated with spoiled fish after inoculation as single and mixed cultures on model fish substrate and storage at 0, 4 and 8 °C. The results indicated a strain-dependent VOC profile that was also affected by the storage temperature. Hierarchical cluster analysis (HCA) successfully grouped the strains based on VOC profile at each studied temperature, while some potential Chemical Spoilage Indices (CSI) were revealed. The findings of the present work will contribute to the understanding of the metabolic activity of particular strains of Pseudomonas and to reveal any potential CSI for rapid evaluation of fish spoilage/freshness status.

Bacterial communities, microbial populations, and antibiotic resistance of potential pathogens in the water and fish (Cyprinus carpio, flesh and gut) from different areas (A1, A2 and A3—A1 was linked with river water, A2 with cattle activity, and A3 with waters of a spring after heavy rains) of Lake Karla (Thessaly, Central Greece) were investigated. The isolated bacteria were identified using Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and were tested for resistance in 21 antibiotics. The microbiota composition of fish flesh was also studied using 16S amplicon-based sequencing Serratia fonticola and several species of Aeromonas (e.g., Aeromonas salmonicida, Aeromonas bestiarium, Aeromonas veronii, etc.) exhibited the highest abundances in all studied samples, while the microbiota profile between the three studied areas was similar, according to the culture-dependent analysis. Of them, S. fonticola was found to be resistant in the majority of the antibiotics for the water and fish (gut and flesh), mainly of the areas A1 and A2. Regarding 16S metabarcoding, the presence of Serratia and Aeromonas at genus level was confirmed, but they found at very lower abundances than those reported using the culture-dependent analysis. Finally, the TVC and the rest of the studied microbiological parameters were found at acceptable levels (4 log cfu/mL or cfu/g and 2–4 log cfu/mL or cfu/g, extremely low levels of E. coli/coliforms) in both water and fish flesh. Based on our findings, the water of Lake Karla would be used for activities such as irrigation, recreation and fishing, however, the development and implementation of a quality management tool for Lake Karla, to ensure environmental hygiene and prevention of zoonosis during the whole year, is imperative.

In the present work, the profiles of bacterial communities of whole and filleted European sea bass (Dicentrarchus labrax), during several storage temperatures (0, 4, 8 and 12 °C) under aerobic and Modified Atmosphere Packaging (MAP) conditions, were examined via the 16S rRNA High-Throughput Sequencing (HTS) approach. Sensorial attributes were also assessed to determine products’ shelf-life. Results indicated that shelf-life was strongly dependent on handling, as well as on temperature and atmosphere conditions. HTS revealed the undisputed dominance of Pseudomonas from the very beginning and throughout storage period in the majority of treatments. However, a slightly different microbiota profile was recorded in MAP-stored fillets at the middle stages of storage, which mainly referred to the sporadic appearance of some bacteria (e.g., Carnobacterium, Shewanella, etc.) that followed the dominance of Pseudomonas. It is noticeable that a major difference was observed at the end of shelf-life of MAP-stored fillets at 12 °C, where the dominant microbiota was constituted by the genus Serratia, while the relative abundance of Pseudomonas and Brochothrix was more limited. Furthermore, at the same temperature under aerobic storage of both whole and filleted fish, Pseudomonas almost co-existed with Acinetobacter, while the presence of both Erwinia and Serratia in whole fish was noteworthy. Overall, the present study provides useful information regarding the storage fate and spoilage status of whole and filleted European sea bass, suggesting that different handling and storage conditions influence the shelf-life of sea bass by favoring or delaying the dominance of Specific Spoilage Organisms (SSOs), affecting in parallel to some extent the formation of their consortium that is responsible for products’ sensorial deterioration. Such findings enrich the current knowledge and should be used as a benchmark to develop specific strategies aiming to delay spoilage and thus increase the products’ added value.

The profiles of bacterial communities and volatile organic compounds (VOCs) of farmed red seabream (Pagrus major) from two batches during ice storage were studied using 16S metabarcoding (culture independent approach) and headspace Solid Phase Micro-Extraction—Gas Chromatography/Mass Spectrometry (SPME-GC/MS) analysis, respectively. Sensory attributes and microbiological parameters were also evaluated. At Day 12 (shelf-life for both batches based on sensory evaluation), using classical microbiological analysis, Total Viable Counts (TVC) were found at the levels of 7–8 log cfu/g, and Pseudomonas and/or H2S producing bacteria dominated. On the other hand, the culture independent 16S metabarcoding analysis showed that Psychrobacter were the most abundant bacteria in fish tissue from batch 1, while Pseudomonas and Psychrobacter (at lower abundance) were the most abundant in fish from batch 2. Differences were also observed in VOC profiles between the two batches. However, combining the VOC results of the two batches, 15 compounds were found to present a similar trend during fish storage. Of them, 2-methylbutanal, 3-methylbutanal, 3-methyl-1-butanol, ethanol, 2,4 octadiene (2 isomers), ethyl lactate, acetaldehyde and (E)-2-penten-1-ol could be used as potential spoilage markers of red seabream because they increased during storage, mainly due to Psychrobacter and/or Pseudomonas activity and/or chemical activity (e.g., oxidation). Additionally, VOCs such as propanoic acid, nonanoic acid, decanoic acid, 1-propanol, 3,4-hexanediol and hexane decreased gradually with time, so they could be proposed as freshness markers of red seabream. Such information will be used to develop intelligent approaches for the rapid evaluation of spoilage course in red seabream during ice storage.

Spoilage status of whole and filleted chill-stored meagre caught in January and July was evaluated using sensory, microbiological, 16S metabarcoding and Volatile Organic Compounds (VOCs) analysis. Based on the sensory analysis, shelf-life was 15 and 12 days for the whole fish taken in January and July, respectively, while 7 days for fish fillets of both months. For the whole fish, Total Viable Counts (TVC) at the beginning of storage was 2.90 and 4.73 log cfu/g for fish caught in January and July respectively, while it was found about 3 log cfu/g in fish fillets of both months. The 16S metabarcoding analysis showed different profiles between the two seasons throughout the storage. Pseudomonas (47%) and Psychrobacter (42.5%) dominated in whole meagre of January, while Pseudomonas (66.6%) and Shewanella (10.5%) dominated in fish of July, at the end of shelf-life. Regarding the fillets, Pseudomonas clearly dominated at the end of shelf-life for both months. The volatile profile of meagre was predominated by alcohols and carbonyl compounds. After univariate and multivariate testing, we observed one group of compounds (trimethylamine, 3-methylbutanoic acid, 3-methyl-1-butanol) positively correlating with time of storage and another group with a declining trend (such as heptanal and octanal). Furthermore, the volatile profile seemed to be affected by the fish culturing season. Our findings provide insights into the spoilage mechanism and give information that helps stakeholders to supply meagre products of a high-quality level in national and international commerce.

In this review we propose the use of telomeric length (TL) as an authenticity marker that could provide an alternative method for differentiating fish and seafood samples or detecting fraud. Considering the ever-growing number of incidents of economically motivated fish and seafood adulteration using even more sophisticated methods to overcome current authenticity markers, the need to identify novel authenticity markers becomes essential. The TL of fish and seafood depends on individual characteristics (e.g., sex, age) and the environmental stimuli (e.g., temperature, water quality) to which these are exposed. Hence, both wild marine and freshwater populations occupying different geographical origin habitats might differ substantially because of the environmental cues affecting them. Moreover, the implementation of various rearing practices in aquaculture, such as different levels of fish and seafood density and increased ambient noise combined with site-specific environmental cues could affect TL, providing regulatory authorities with valuable information by distinguishing wild from reared populations and organic from conventional ones. In the present review the effects of both the environmental conditions and individual characteristics on the telomeric stability of fish and seafood telomeres are discussed, suggesting TL as a potential prospect authenticity marker that could be used to prevent fish and seafood adulteration. [ABSTRACT FROM AUTHOR]

The cultivable microbiota isolated from three sea bass products (whole, gutted, and filleted fish from the same batch) during chilled storage and the effect of primary processing on microbial communities in gutted and filleted fish were studied. Microbiological and sensory changes were also monitored. A total of 200 colonies were collected from TSA plates at the beginning and the end of fish shelf-life, differentiated by High Resolution Sequencing (HRM) and identified by sequencing analysis of the V3–V4 region of the 16S rRNA gene. Pseudomonas spp. followed by potential pathogenic bacteria were initially found, while Pseudomonasgessardii followed by other Pseudomonas or Shewanella species dominated at the end of fish shelf-life. P. gessardii was the most dominant phylotype in the whole sea bass, P. gessardii and S. baltica in gutted fish, while P. gessardii and P. fluorescens were the most dominant bacteria in sea bass fillets. To conclude, primary processing and storage affect microbial communities of gutted and filleted fish compared to the whole fish. HRM analysis can easily differentiate bacteria isolated from fish products and reveal the contamination due to handling and/or processing, and so help stakeholders to immediately tackle problems related with microbial quality or safety of fish.

A literature search and systematic review were conducted to present and discuss the most recent research studies for the past twenty years on the application of non-thermal methods for ensuring the microbiological safety and quality of fish and seafood. This review presents the principles and reveals the potential benefits of high hydrostatic pressure processing (HHP), ultrasounds (US), non-thermal atmospheric plasma (NTAP), pulsed electric fields (PEF), and electrolyzed water (EW) as alternative methods to conventional heat treatments. Some of these methods have already been adopted by the seafood industry, while others show promising results in inactivating microbial contaminants or spoilage bacteria from solid or liquid seafood products without affecting the biochemical or sensory quality. The main applications and mechanisms of action for each emerging technology are being discussed. Each of these technologies has a specific mode of microbial inactivation and a specific range of use. Thus, their knowledge is important to design a practical application plan focusing on producing safer, qualitative seafood products with added value following today’s consumers’ needs.

The aim of the present study was to assess the microbiological quality of farmed sea bass (Dicentrarchus labrax) fillets stored under aerobic conditions and modified atmosphere packaging (MAP) (31% CO2, 23% O2, 46% Ν2,) at 0, 4, 8, and 12 °C using Fourier transform infrared (FTIR) spectroscopy and multispectral imaging (MSI) in tandem with data analytics, taking into account the results of conventional microbiological analysis. Fish samples were subjected to microbiological analysis (total viable counts (TVC), Pseudomonas spp., H2S producing bacteria, Brochothrix thermosphacta, lactic acid bacteria (LAB), Enterobacteriaceae, and yeasts) and sensory evaluation, together with FTIR and MSI spectral data acquisition. Pseudomonas spp. and H2S-producing bacteria were enumerated at higher population levels compared to other microorganisms, regardless of storage temperature and packaging condition. The developed partial least squares regression (PLS-R) models based on the FTIR spectra of fish stored aerobically and under MAP exhibited satisfactory performance in the estimation of TVC, with coefficients of determination (R2) at 0.78 and 0.99, respectively. In contrast, the performances of PLS-R models based on MSI spectral data were less accurate, with R2 values of 0.44 and 0.62 for fish samples stored aerobically and under MAP, respectively. FTIR spectroscopy is a promising tool to assess the microbiological quality of sea bass fillets stored in air and under MAP that could be effectively employed in the future as an alternative method to conventional microbiological analysis.

The aquatic environment is a reservoir of many species that have not yet been exploited at a global level and have not been extensively investigated. The aim of the present work was to study the microbial populations, the bacterial communities and physicochemical parameters (pH, water activity, humidity, salinity) in raw, frozen, boiled, dehydrated and salted final products of two sea cucumber species (Holothuria polii and Holothuria tubulosa) originating from the Hellenic seawaters. The results indicated that all products were found at acceptable levels from a microbiological point of view. The metabarcoding analysis of the 16S rRNA gene revealed the existence of several different bacterial groups, the presence and abundance of which were mainly dependent on product type, even though some moderate differences in the microbiota profile between the two sea cucumber species were also detected in some of the products. Overall, the present work deals with an underexplored aquatic product and provides novel and useful information for the aquatic food industry, consumers and other stakeholders, increasing, in parallel, the need for further scientific attention in the near future. Our findings could be exploited as a baseline to highlight a promising aquatic food product provided to the international market. [ABSTRACT FROM AUTHOR]

Fish spoilage occurs due to production of metabolites during storage, from bacterial action and chemical reactions, which leads to sensory rejection. Investigating the volatilome profile can reveal the potential spoilage markers. The evolution of volatile organic molecules during storage of European seabass (Dicentrarchus labrax) fillets and Atlantic salmon (Salmo salar) slices under modified atmosphere packaging at 2 °C was recorded by solid-phase microextraction combined with gas chromatography-mass spectrometry. Total volatile basic nitrogen (TVB-N), microbiological, and sensory changes were also monitored. The shelf life of seabass fillets and salmon slices was 10.5 days. Pseudomonas and H2S-producing bacteria were the dominant microorganisms in both fish. TVB-N increased from the middle of storage, but never reached concentrations higher than the regulatory limit of 30–35 mg N/100 g. The volatilome consisted of a number of aldehydes, ketones, alcohols and esters, common to both fish species. However, different evolution patterns were observed, indicating the effect of fish substrate on microbial growth and eventually the generation of volatiles. The compounds 3-hydroxy-2-butanone, 2,3-butanediol, 2,3-butanedione and acetic acid could be proposed as potential spoilage markers. The identification and quantification of the volatilities of specific fish species via the development of a database with the fingerprint of fish species stored under certain storage conditions can help towards rapid spoilage assessment.

The aim of the present work was to evaluate the effect of various hurdles such as aw and pH as well as the storage atmosphere on the microbiological and sensory changes of minimally processed (lightly brined or marinated with acetic or citric acid) European sea bass (Dicentrarchus labrax) fillets. The results indicated that the shelf-life of brined fillets stored aerobically was 6 d while that of vacuum and MAP-stored was 12 d, since a reduced growth rate of spoilage bacteria was recorded. The physicochemical characteristics such as aw and water phase salt (WPS) were not considerably changed, while the oxygen levels into the packages ensure the microbiological safety of the product. The fillets marinated with acetic acid exhibited a longer shelf-life at 30 and 40 d under aerobic and reduced oxygen conditions, respectively, while the products marinated with citric acid were at 25 and 35 d respectively. A low pH resulted in reduced or even limited microbial levels, especially for the fillets marinated with acetic acid; something that ensures microbiological safety as well. The low or limited microbial levels in conjugation with the sensory attributes indicated that spoilage may be due to other mechanisms such as autolysis rather than microbial activity. Overall, the present work highlights the potential for further research and development of minimally processed, microbiologically safe and stable with extended shelf-life value added seafood to satisfy the corresponding consumer demands. [ABSTRACT FROM AUTHOR]

This study aimed to develop a better understanding of organic sea bream aquaculture production in Greece, in particular its consequences for fish growth, health and body composition, and to propose and update standards for sustainable organic sea bream farming. Gilthead sea bream were kept in sea cages at densities of 4 kg m–3 (organic) and 15 kg m–3 (conventional), and were fed organically produced feed (45% crude protein, 14% fat) or conventional feed (46% crude protein, 17% fat). The amino acid profile of the conventional diet, particularly the lysine content, which is one of most important dietary amino acids for sea bream, appeared to be unsatisfactory. “Organic” sea bream stored less fat content in their white muscle than the conventional sea bream. The liver lipid content was lower and the hepatosomatic index was higher for the organic sea bream. The microbiological analysis showed that both Enterobacteriaceae and Escherichia coli on the skin were below the enumeration detection limit in both the organic and conventional sea bream. Total viable counts on the skin and muscle of both the organically and conventionally cultured sea bream were approximately 3 log cfu g –1, which is well below the acceptable limit (7 log cfu g–1) for marine species. The results showed that the combination of a low stocking density and feed with a different ingredient composition but similar nutritional value resulted in similar growth rates and nutrient profiles of the final product. Further research on nutrition is required to provide information on setting the appropriate standards for organic sea bream aquaculture to ensure that the final product is in line with the consumers’ preferences.

Water-methanol extract from Filipendula ulmaria contains a variety of phenolic compounds, such as caffeic, p-coumaric and vanillic acid, myricetin, etc, which demonstrate antibacterial activity. Monitoring this activity in the broth using absorbance measurements showed that species of the Enterobacteriaceae family were more resistant than other Gram-negative and Gram-positive bacteria tested. Acidic environment enhanced the antibacterial activity of Filipendula ulmaria extract when it was tested against Salmonella Enteritidis PT4 and Listeria monocytogenes Scott A. The efficacy of Filipendula ulmaria extract against selected foodborne psychrotrophic bacteria was also tested using solid laboratory media and low incubation temperatures for better simulation of food preservation conditions. Higher concentrations of the extract, compared to minimum inhibitory concentration determined in the broth, were needed for satisfactory inhibition of spoilage bacteria. Potential use of Filipendula ulmaria extract as natural food preservative was also examined against natural spoilage flora and inoculated pathogenic bacteria on fish flesh and fish roe product (tarama salad). No significant differences of viable populations of spoilage or pathogenic bacteria were found between the treated samples and controls. Further trials of Filipendula ulmaria extract should be carried out in acidic foods with low fat and protein content, supplemented with additional adjuncts, in order to explore its potential as effective natural food antimicrobial agent.

Aromatic plants contain natural antioxidant constituents such as phenolic compounds, which have attracted a great deal of public and scientific interest because of their health-promoting effects as antioxidants. Five plants, Filipendula ulmaria (meadow sweet), Crataegus monogyna (hawthorn), Polygonum aviculare (polygonum), Potentilla anserina (silverweed),and Pelargonium purpureum (little robin), have been examined in order to determine their phenolic composition. Reversed-phase high performance liquid chromatography (RP-HPLC) was employed for the identification and quantification of phenolic compounds ofthe aforementioned plants. Gas chromatography–mass spectrometry method (GC-MS) was also used for identification of phenolic compounds after silylation. Analysis of the non-volatile and thermolabile phenolic compounds by GC-MS presupposes their conversion intovolatile and thermotolerant derivatives. The derivatization process was optimized againstreagents, temperature and reaction time. The antioxidant capacity was determined in driedplants and in their methanol extracts with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenoliccontent in the extracts was determined spectrometrically applying the Folin-Ciocalteu assay and it ranged from 7.2 to 28.2 gallic acid equivalents (GAE)/(mg/mL). Antimicrobial activity of the extracts against selected microorganisms was performed using the disk diffusion method. Gram-(+) bacteria were more sensitive to the plant extracts than Gram-(–) bacteria.

Microbial spoilage is the main cause of quality deterioration in seafood. Several strains of psychotropic Pseudomonas have been found to dominate in such products, producing a plethora of volatile organic compounds (VOC). Herein, we investigated the growth of and VOC production by seven strains of Pseudomonas associated with spoiled fish after inoculation as single and mixed cultures on model fish substrate and storage at 0, 4 and 8 °C. The results indicated a strain-dependent VOC profile that was also affected by the storage temperature. Hierarchical cluster analysis (HCA) successfully grouped the strains based on VOC profile at each studied temperature, while some potential Chemical Spoilage Indices (CSI) were revealed. The findings of the present work will contribute to the understanding of the metabolic activity of particular strains of Pseudomonas and to reveal any potential CSI for rapid evaluation of fish spoilage/freshness status. [ABSTRACT FROM AUTHOR]

Bacterial communities, microbial populations, and antibiotic resistance of potential pathogens in the water and fish (Cyprinus carpio, flesh and gut) from different areas (A1, A2 and A3—A1 was linked with river water, A2 with cattle activity, and A3 with waters of a spring after heavy rains) of Lake Karla (Thessaly, Central Greece) were investigated. The isolated bacteria were identified using Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) and were tested for resistance in 21 antibiotics. The microbiota composition of fish flesh was also studied using 16S amplicon-based sequencing Serratia fonticola and several species of Aeromonas (e.g., Aeromonas salmonicida, Aeromonas bestiarium, Aeromonas veronii, etc.) exhibited the highest abundances in all studied samples, while the microbiota profile between the three studied areas was similar, according to the culture-dependent analysis. Of them, S. fonticola was found to be resistant in the majority of the antibiotics for the water and fish (gut and flesh), mainly of the areas A1 and A2. Regarding 16S metabarcoding, the presence of Serratia and Aeromonas at genus level was confirmed, but they found at very lower abundances than those reported using the culture-dependent analysis. Finally, the TVC and the rest of the studied microbiological parameters were found at acceptable levels (4 log cfu/mL or cfu/g and 2–4 log cfu/mL or cfu/g, extremely low levels of E. coli/coliforms) in both water and fish flesh. Based on our findings, the water of Lake Karla would be used for activities such as irrigation, recreation and fishing, however, the development and implementation of a quality management tool for Lake Karla, to ensure environmental hygiene and prevention of zoonosis during the whole year, is imperative. [ABSTRACT FROM AUTHOR]

Marine environment disposes a plethora of bioactive compounds with unique properties and remarkable potential for biotechnological applications. A lot of those compounds can be used by the food industry as natural preservatives, pigments, stabilizers, gelling agents, etc., while others exhibits beneficial effects and can be used as functional food ingredients, nutraceuticals, dietary supplements and prebiotics. Interdisciplinary approach is required to increase our knowledge, explore the potential of marine environment and produce value-added food for all.

In the present work, the profiles of bacterial communities of whole and filleted European sea bass (Dicentrarchus labrax), during several storage temperatures (0, 4, 8 and 12 °C) under aerobic and Modified Atmosphere Packaging (MAP) conditions, were examined via the 16S rRNA High-Throughput Sequencing (HTS) approach. Sensorial attributes were also assessed to determine products' shelf-life. Results indicated that shelf-life was strongly dependent on handling, as well as on temperature and atmosphere conditions. HTS revealed the undisputed dominance of Pseudomonas from the very beginning and throughout storage period in the majority of treatments. However, a slightly different microbiota profile was recorded in MAP-stored fillets at the middle stages of storage, which mainly referred to the sporadic appearance of some bacteria (e.g., Carnobacterium, Shewanella, etc.) that followed the dominance of Pseudomonas. It is noticeable that a major difference was observed at the end of shelf-life of MAP-stored fillets at 12 °C, where the dominant microbiota was constituted by the genus Serratia, while the relative abundance of Pseudomonas and Brochothrix was more limited. Furthermore, at the same temperature under aerobic storage of both whole and filleted fish, Pseudomonas almost co-existed with Acinetobacter, while the presence of both Erwinia and Serratia in whole fish was noteworthy. Overall, the present study provides useful information regarding the storage fate and spoilage status of whole and filleted European sea bass, suggesting that different handling and storage conditions influence the shelf-life of sea bass by favoring or delaying the dominance of Specific Spoilage Organisms (SSOs), affecting in parallel to some extent the formation of their consortium that is responsible for products' sensorial deterioration. Such findings enrich the current knowledge and should be used as a benchmark to develop specific strategies aiming to delay spoilage and thus increase the products' added value. [ABSTRACT FROM AUTHOR]

Spoilage status of whole and filleted chill-stored meagre caught in January and July was evaluated using sensory, microbiological, 16S metabarcoding and Volatile Organic Compounds (VOCs) analysis. Based on the sensory analysis, shelf-life was 15 and 12 days for the whole fish taken in January and July, respectively, while 7 days for fish fillets of both months. For the whole fish, Total Viable Counts (TVC) at the beginning of storage was 2.90 and 4.73 log cfu/g for fish caught in January and July respectively, while it was found about 3 log cfu/g in fish fillets of both months. The 16S metabarcoding analysis showed different profiles between the two seasons throughout the storage. Pseudomonas (47%) and Psychrobacter (42.5%) dominated in whole meagre of January, while Pseudomonas (66.6%) and Shewanella (10.5%) dominated in fish of July, at the end of shelf-life. Regarding the fillets, Pseudomonas clearly dominated at the end of shelf-life for both months. The volatile profile of meagre was predominated by alcohols and carbonyl compounds. After univariate and multivariate testing, we observed one group of compounds (trimethylamine, 3-methylbutanoic acid, 3-methyl-1-butanol) positively correlating with time of storage and another group with a declining trend (such as heptanal and octanal). Furthermore, the volatile profile seemed to be affected by the fish culturing season. Our findings provide insights into the spoilage mechanism and give information that helps stakeholders to supply meagre products of a high-quality level in national and international commerce. [ABSTRACT FROM AUTHOR]

The EuroCigua project main objective is to characterize the risk of Ciguatera Poisoning (CP) in Europe including several specific objectives: to determine the incidence of ciguatera in Europe and the epidemiological characteristics of cases; to assess the presence of ciguatoxin in food and the environment in Europe and to develop and validate methods for the detection, quantification and confirmation of the presence of ciguatoxin contaminated specimens. This report compiles the activities carried out during the EuroCigua project from the signing in April 2016 until December 2020. The present document corresponds to Deliverable No. 6: "Final Scientific Report" on Risk characterization of ciguatera food poisoning in Europe of the Specific Agreement no. 1 "MANAGEMENT AND SCIENTIFIC COORDINATION" within the Framework Partnership Agreement GP/EFSA/AFSCO/2015/03 "Risk characterization of ciguatera food poisoning in Europe". [ABSTRACT FROM AUTHOR]

The present document corresponds to the final report (deliverable No. 7) of the specific agreement (SA 2) "To determine the incidence and epidemiological characteristics of ciguatera cases in Europe" within the Framework Partnership Agreement GP/EFSA/AFSCO/2015/03 "Risk characterization of ciguatera food poisoning in Europe". This report summarises the activities performed, results and conclusions obtained in the specific agreement. The objective of the SA2 was to estimate the incidence and to describe the epidemiological characteristics of ciguatera in the EU/EEA. A protocol for a harmonized ciguatera surveillance in the European Union (EU)/Economic European Area (EEA) was elaborated. The protocol includes a ciguatera case definition and questionnaires for collecting information on ciguatera cases or outbreak. Around half of the countries answered the data call for reporting cases. Four countries reported 34 outbreaks from 2012 to 2019. Spain and Portugal reported outbreaks due to consumption of fish captured in the Canary Islands and Madeira (autochthonous outbreaks), mainly due to consumption of Seriola spp. and Ephinephelus spp. In more than half of these outbreaks, the fish was captured by sport fishing. Germany and France reported outbreaks due to consumption of fish imported from outside the EU (imported outbreaks, mainly Lutjanus spp). Spain, Germany and France reported outbreaks in travellers to tropical endemic territories (travel related outbreaks). All the outbreaks cases presented neurological symptoms, most of them had also gastrointestinal symptoms and few outbreak cases mentioned cardiovascular symptoms. Six countries (Austria, France, Germany, Portugal, Spain and Switzerland) reported 34 single cases. The incidence rate in the EU/EEA was very low (0.0054 cases per 100.000 inhabitants per year). The incidence rate in the Canary Islands was higher 0.47 cases/100.000 inhabitants. [ABSTRACT FROM AUTHOR]

The verification of authenticity and detection of food mislabeling are elements that have been of high importance for centuries. During the last few decades there has been an increasing consumer demand for the verification of food identity and the implementation of stricter controls around these matters. Fish and seafood are among the most easily adulterated foodstuffs mainly due to the significant alterations of the species' morphological characteristics that occur during the different types of processing, which render the visual identification of the animals impossible. Even simple processes, such as filleting remove very important morphological elements and suffice to prevent the visual identification of species in marketed products. Novel techniques have therefore been developed that allow identification of species, the differentiation between species and also the differentiation of individuals that belong to the same species but grow in different populations and regions. Molecular markers have been used during the last few decades to fulfill this purpose and several improvements have been implemented rendering their use applicable to a commercial scale. The reliability, accuracy, reproducibility, and time‐and cost‐effectiveness of these techniques allowed them to be established as routine methods in the industry and research institutes. This review article aims at presenting the most important molecular markers used for the authentication of fish and seafood. The most important techniques are described, and the results of numerous studies are outlined and discussed, allowing interested parties to easily access and compare information about several techniques and fish/seafood species. [ABSTRACT FROM AUTHOR]

A literature search and systematic review were conducted to present and discuss the most recent research studies for the past twenty years on the application of non-thermal methods for ensuring the microbiological safety and quality of fish and seafood. This review presents the principles and reveals the potential benefits of high hydrostatic pressure processing (HHP), ultrasounds (US), non-thermal atmospheric plasma (NTAP), pulsed electric fields (PEF), and electrolyzed water (EW) as alternative methods to conventional heat treatments. Some of these methods have already been adopted by the seafood industry, while others show promising results in inactivating microbial contaminants or spoilage bacteria from solid or liquid seafood products without affecting the biochemical or sensory quality. The main applications and mechanisms of action for each emerging technology are being discussed. Each of these technologies has a specific mode of microbial inactivation and a specific range of use. Thus, their knowledge is important to design a practical application plan focusing on producing safer, qualitative seafood products with added value following today's consumers' needs. [ABSTRACT FROM AUTHOR]

The growth and activity of Shewanella putrfaciens, Brochothrix thermosphacta and pseudomonas species when cultured individually or in all possible combinations in gel cassettes system supplemented with 0.1 percent glucose at 5 degree Celsius were investigated. The findings stated that the coexistence of these microorganisms affected not only each growth rate but also their type of metabolic end products compared to the control cultures.

Abstract: BACKGROUND: Aromatic plants may be contaminated with a wide range of microorganisms, making them a potential health hazard when infused or added to ready‐to‐eat meals. To ensure safety, the effect of gaseous ozone treatment on the population of aerobic plate counts (APC), hygienic indicators (Escherichia coli, Enterococcus spp. and Enterobacteriaceae) and fungi was investigated for five dried aromatic plants: oregano, thyme, mountain tea, lemon verbena and chamomile. Selection, isolation and further fungi identification were based on the phenotypic and macro‐ and microscopic characteristics. RESULTS: Prior to ozonation, APC on five dried aromatic plants was in the range 5–7 log colony‐forming units (CFU) g–1. The APC exhibited a 4 log reduction, from around 6.5 to 2.5 in the case of oregano, and only a 1–2 log reduction for other herbs after 30 or 60 min of 4 ppm gaseous ozone treatment. Enterococcus spp. and E. coli were not detected on any of the tested dried aromatic plants. The fungi counts were 2–4 log CFU g–1 before ozonation. Aspergillus spp, Penicillium spp, Cladosporium spp, Alternaria spp, Fusarium spp., Ulocladium spp. and some unknown fungi were detected on plants before ozone treatment. Aspergillus spp. and/or Penicillium spp. were only detected on mountain tea and thyme plant material after 60 min of ozonation. CONCLUSION: The present study provides information about the efficiency of ozone on the microbial decontamination of dried aromatic plants. Treatment with gaseous ozone at 4 ppm for 30 min in the case of dried oregano and 60 min in the case of chamomile and lemon verbena could be used as alternative disinfection methods. © 2017 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Abstract: As a tool for stock identification the body morphology and the free amino acid (FAA) profile variability were determined among farmed rainbow trout Oncorhynchus mykiss (Walbaum 1792) from different geographical areas throughout Greece, using geometric morphometrics and Reversed Phase‐High‐Performance Liquid Chromatography. Significant intraspecific body shape variation was detected among the fish stocks of different origin. Canonical Variate Analysis distinguished populations into two groups being consistent to a great extent with a previous population genetics study. General linear models did not support an association of haplotypes and diet with shape. However, shape was significantly correlated with origin, water temperature, geological‐chemical‐climatic zones, histidine and arginine. Therefore, morphological variation was primarily driven by environmentally induced differences among these zones being the result of phenotypic plasticity. Environmental cues and rearing conditions played the pivotal role in comparison to broodstock origin. Specimens presented different FAA profiles depending on their geographical origin. Principal Component Analysis showed a division between eastern and western Greece which might be owed to the existence of climatic fluctuations affecting the abiotic factors. This combined approach offers an important tool for stock identification thus assisting environmental risk assessments in evaluation of potential ecological effects and fostering research excellence in sustainable aquaculture management. [ABSTRACT FROM AUTHOR]

Exploration of initial and spoilage microbiota grown on plates of whole sea bream stored aerobically at 0 (ice), 5 and 15 °C, was conducted by 16S rRNA gene sequence analysis. The course of spoilage was recorded by monitoring microbiological, sensory and chemical changes. Shelf-life of sea bream determined by sensory assessment was 16, 5 and 2 days at 0 (ice), 5 and 15 ο C, respectively. Pseudomonas spp. was the dominant spoilage population of whole sea bream at all temperatures tested. A sum of 144 colonies were isolated from TSA (Tryptone Soy Agar) plates and identified by genotypic approach at the beginning and at the sensory rejection time points. Regarding initial microbiota, Pseudomonas fragi was the most abundant compared to the rest bacteria ( Pseudomonas fluorescens , Enterobacter hormaechei , Chryseobacterium carnipullorum ). P. fragi was also the dominant microorganism of fish stored at 0 and 5, while P. fluorescens at 15 ο C. Concluding, genotypic approach gives accurate identification of the dominant spoilage microorganisms providing us with valuable information regarding microbiological spoilage of fish. [ABSTRACT FROM AUTHOR]

BACKGROUND Sea bream fillets are one of the most important value-added products of the seafood market. Fresh seafood spoils mainly owing to bacterial action. In this study an exploration of initial and spoilage microbiota of sea bream fillets stored under air and commercial modified atmosphere packaging ( MAP) at 0 and 5 °C was conducted by 16S rRNA gene sequence analysis of isolates grown on plates. Sensory evaluation and enumeration of total viable counts and spoilage microorganisms were also conducted to determine shelf life and bacterial growth respectively. RESULTS Different temperatures and atmospheres affected growth and synthesis of spoilage microbiota as well as shelf life. Shelf life under air at 0 and 5 °C was 14 and 5 days respectively, while under MAP it was 20 and 8 days respectively. Initial microbiota were dominated by Pseudomonas fluorescens, Psychrobacter and Macrococcus caseolyticus. Different temperatures and atmospheres affected the synthesis of spoilage microbiota. At the end of shelf life, different phylotypes of Pseudomonas closely related to Pseudomonas fragi were found to dominate in most cases, while Pseudomonas veronii dominated in fillets under MAP at 0 °C. Furthermore, in fillets under MAP at 5 °C, new dominant species such as Carnobacterium maltaromaticum, Carnobacterium divergens and Vagococcus fluvialis were revealed. CONCLUSION Different temperature and atmospheric conditions affected bacterial growth, shelf life and the synthesis of spoilage microbiota. Molecular identification revealed species and strains of microorganisms that have not been reported before for sea bream fillets stored under various conditions, thus providing valuable information regarding microbiological spoilage. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

BACKGROUND An increasing interest in snail farming in Greece and other European countries has been observed. Despite the fact that edible snails have been involved with problems of Salmonella spp. contamination, there are to our knowledge only limited studies regarding microbiological safety and hygiene of such products. Enumeration of microbial populations and presence/absence of Salmonella spp. in snail meat and intestines of wild Cornu aspersum, Helix lucorum and cultured Cornu aspersum snails from indoor/outdoor type farms was conducted. Furthermore, snail-processing steps were simulated in the laboratory and the population reduction in snail meat was determined. RESULTS Microbial populations were higher in intestines than snail meat in almost all cases. Escherichia coli/coliforms and Enterococcus spp. populations were lower in the intestines and snail meat of cultured C. aspersum. Salmonella spp. were detected in the intestines and snail meat of wild snails only. The high levels of bacterial populations were considerably reduced after the appropriate processing. CONCLUSION The lower populations of E. coli/coliforms, Enterococcus spp. and especially the absence of Salmonella spp. in cultured snails show that the controlled conditions decrease the possibility of pathogen presence and contribute to food safety and public health. © 2013 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The profiles of bacterial communities and volatile organic compounds (VOCs) of farmed red seabream (Pagrus major) from two batches during ice storage were studied using 16S metabarcoding (culture independent approach) and headspace Solid Phase Micro-Extraction—Gas Chromatography/Mass Spectrometry (SPME-GC/MS) analysis, respectively. Sensory attributes and microbiological parameters were also evaluated. At Day 12 (shelf-life for both batches based on sensory evaluation), using classical microbiological analysis, Total Viable Counts (TVC) were found at the levels of 7–8 log cfu/g, and Pseudomonas and/or H2S producing bacteria dominated. On the other hand, the culture independent 16S metabarcoding analysis showed that Psychrobacter were the most abundant bacteria in fish tissue from batch 1, while Pseudomonas and Psychrobacter (at lower abundance) were the most abundant in fish from batch 2. Differences were also observed in VOC profiles between the two batches. However, combining the VOC results of the two batches, 15 compounds were found to present a similar trend during fish storage. Of them, 2-methylbutanal, 3-methylbutanal, 3-methyl-1-butanol, ethanol, 2,4 octadiene (2 isomers), ethyl lactate, acetaldehyde and (E)-2-penten-1-ol could be used as potential spoilage markers of red seabream because they increased during storage, mainly due to Psychrobacter and/or Pseudomonas activity and/or chemical activity (e.g., oxidation). Additionally, VOCs such as propanoic acid, nonanoic acid, decanoic acid, 1-propanol, 3,4-hexanediol and hexane decreased gradually with time, so they could be proposed as freshness markers of red seabream. Such information will be used to develop intelligent approaches for the rapid evaluation of spoilage course in red seabream during ice storage. [ABSTRACT FROM AUTHOR]

BACKGROUND: Fresh fish and seafoods are very perishable products mainly owing to microbial activity of specific spoilage micro-organisms. Application of hurdle technology leads to a variety of processed products with extended shelf life. In this study, sensory evaluation and microbiological analysis were carried out on 17 processed seafood products stored at 4 °C to determine their shelf life and the predominant spoilage micro-organisms. RESULTS: Shelf life determined by sensory analysis varied from 66 to 180 days depending on the product. The cause of spoilage for most of the products was the development of off-flavours/off-odours, while two products were rejected owing to oil discolouration. Pseudomonads were in most cases below detection limit. H2S-producing bacteria, Brochothrix thermosphacta and Enterobacteriaceae were below detection limit throughout the experiment. The predominant spoilage micro-organisms were lactic acid bacteria and yeasts. Hygiene indicators such as Staphylococcus spp. and total coliforms were also below detection limit in all samples. CONCLUSION: Primarily the initial pH and secondarily the NaCl content determined shelf life duration. Under the applied conditions, lactic acid bacteria and yeasts predominated. The contribution of chemical oxidation and/or autolysis to spoilage and shelf life might be important for most of the products. © 2012 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

Water-methanol extract from Filipendula uhnaria contains a variety of phenolic compounds, such as caffeic, p-coumaric and vanillic acid, myricetin, etc, which demonstrate multibacterial activity. Monitoring this activity in the broth using absorbance measurements showed that species of the Enterobacteriaceae family were more resistant than other Gram-negative and Gram-positive bacteria tested. Acidic environment enhanced the antibacterial activity of Filipendula ulmaria extract when it was tested against Salmonella Enteritidis PT4 and Listeria monocytogenes Scott A. The efficacy of Filipendula ulmaria extract against selected foodborne psychrotrophic bacteria was also tested using solid laboratory media and low incubation temperatures for better simulation of food preservation conditions. Higher concentrations of the extract, compared to minimum inhibitory concentration determined in the broth, were needed for satisfactory inhibition of spoilage bacteria. Potential use of Filipendula ulmaria extract as natural food preservative was also examined against natural spoilage flora and inoculated pathogenic bacteria on fish flesh and fish roe product (tarama salad). No significant differences of viable populations of spoilage or pathogenic bacteria were found between the treated samples and controls. Further trials of Filipendula ulmaria extract should be carried out in acidic foods with low fat and protein content, supplemented with additional adjuncts, in order to explore its potential as effective natural food antimicrobial agent. [ABSTRACT FROM AUTHOR]

High pressure processing (HPP) at ultra-low temperatures was conducted against Listeria monocytogenes and Salmonella enterica in frozen pink salmon fillets. Quality changes, such as drip loss, color and odor attributes were recorded in non-inoculated pollock, pink salmon and tuna fillets. Pressures at 250 and 400 MPa were applied from 0.5 to 10 min. Reductions up to 3.5 log cfu/g were recorded for the treatments performed at −32 °C, in contrast to −50 °C where the reductions were only up to 1.5 log cfu/g. Higher pressure did not cause higher reduction. It was apparent that the main factor contributing to the bacterial inactivation is the phase transition of ice structure from I to III, in contrast to transition from I to II. Drip loss was not higher than the expected with HPP at temperatures above 0 °C, while color changes were negligible. Finally, the odor evaluation did not exhibit considerable differences between untreated and treated samples. High pressure processing at ultra-low temperatures is a promising treatment for bacterial inactivation and retention of quality attributes of frozen fish. Treatment at 250 MPa for only 3 min at temperatures just below −22 °C, which is feasible and affordable, caused a more than 3-log reduction against Listeria monocytogenes and Salmonella enterica , without affecting considerably the quality properties. Thus, the application of low pressure and shorter processing times gives a great potential for industrial application for frozen fish or fish that wouldn't be undesirable to freeze before pressurization. • Pressure at 400 MPa did not cause higher Salmonella and Listeria reduction compared to 250 MPa. • The treatment at −32 °C caused higher log reduction compared to −50 °C. • Pressure of 250 MPa for only 3 min at −32 °C caused a 3-log reduction for both Salmonella and Listeria. • Pressure of 250 MPa for only 3 min at −32 °C did not affect considerably color and odor. [ABSTRACT FROM AUTHOR]

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Abstract: The effect of nisin on growth boundaries of Listeria monocytogenes Scott A in Tryptone Soy Broth (TSB) under different a ws, pH, and temperatures was studied. Growth/no growth turbidity data was modeled using logistic regression. Combinations of various temperatures (5–35°C), pH (4.05–6.70) adjusted with HCl, a ws (0.937–0.998) NaCl (0.5–10.5%) and nisin (0–100IU/ml) were used to monitor the growth/no growth response of L. monocytogenes Scott A for 60 days. The concordance of the logistic regression model was 99.4%, indicating successful data fitting. The minimum pH at which growth was observed was 4.81 at the temperature range of 25–35°C and at a w as high as 0.992. Growth was observed at a w as low as 0.937, at pH 6.7, at the temperature range of 25–35°C. Increasing nisin concentrations above 25IU/ml resulted in a more inhibitory environment for L. monocytogenes. Presence of 100IU/ml resulted in a minimum pH for growth at 5.20, and a minimum a w at 0.967 at the temperature range of 25–35°C. It was remarkable that low to medium salt concentrations (2.5–4.5 NaCl% w/v) provided a protective effect against inhibition of L. monocytogenes by nisin. The present study points out the applicability of growth/no growth modeling in order to study any interactions between various factors affecting initiation of growth of micro-organisms, in which its turn helps the understudying of microbe–food ecosystem relations and the development of safer food. [Copyright &y& Elsevier]

Abstract: The phenolic fraction of plant extracts has been linked to their antioxidant capacity and antimicrobial activity. In the present paper, RP-HPLC with UV detection was employed to distinguish phenolic compounds from the other constituents of the plants examined. The phenolic substances were identified and quantified after comparison with reference standards. A GC–MS method is also presented for characterization of different phenolics as trimethylsilyl derivatives. The antioxidant capacity was determined, in dried plants and in their methanol extracts, with the Rancimat test using sunflower oil as substrate. Both pulverized plants and extracts showed antioxidant capacity. Total phenolic content in the extracts was determined spectrometrically applying the Folin-Ciocalteu assay. It ranged from 2.9 to 28.2 mg gallic acid/100 g dry sample. Antimicrobial activity of the extracts against selected microorganisms was also investigated. [Copyright &y& Elsevier]

Green-table olives, unheated and pasteurized, of Greek cultivar Conservolea were supplemented with glucose or sucrose in various amounts inoculated with Lactobacillus plantarum and fermentation was conducted. Although sugar supplements did not affect the lactic acid bacteria growth rate, they increased the rate of pH drop and the production of acids and lowered the final pH values in all cases. Adequate supplements of sugars (0.5%, 1.0% w v-1), of both types, in unheated olive fermentation, resulted in a fast pH drop, causing a satisfactory halt of Enterobacteriaceae growth in the first days of fermentation, and a subsequent population decline occurred in the following days, eliminating the danger of early stage spoilage and ensuring the safety of the final product. In pasteurized olives, where L. plantarum starter was the only microbial flora, it was considered worth evaluating the fermentation progress without any competition. The rate of pH drop was not as high as in unheated olives, but final pH values and acid development were more pronounced. Lactic acid was the predominant acid developed in both unheated and pasteurized olive fermentation, and increased sugar supplements resulted in faster production and higher yields of this acid. Acetic acid was also produced in low amounts at the end of fermentation, except in the cases of pasteurized olive fermentation supplemented with sucrose, where acetic acid was absent. A switch from homo-fermentative to hetero-fermentative metabolism might have taken place in case of glucose presence, but the same was not observed for sucrose. [Copyright &y& Elsevier]