Carnobacterium Divergens Research Articles

Monitoring of volatile production in cooked poultry products using selected ion flow tube-mass spectrometry

Cooked poultry products are nutritious and economically valuable products that are at risk of bacterial spoilage, which can be postponed by cooling and modified-atmosphere-packaging (MAP). In this study, a cooked chicken product was stored at three different temperature ranges (4–6 °C, 7–9 °C, and 11–13 °C) and volatile production was measured over time using selected ion flow tube-mass spectrometry (SIFT-MS). The identities of the volatiles formed were confirmed by headspace solid-phase microextraction coupled with gas chromatography time-of-flight mass spectrometry (HS-SPME-GC-TOF-MS) analysis. In total, 33 volatiles were proposed using the latter technique and their concentrations were calculated using product ion counts after assignment of these counts to specific volatiles. The results indicated that 1-octen-3-ol, 2,3-butanediol, acetoin, benzaldehyde, ethanol, methylbutanal, and methylbutanol may serve as biomarkers for bacterial growth and/or chemical degradation of cooked poultry products. In parallel, the bacterial loads of the product samples were determined on selective agar media. A total of 495 bacterial isolates was classified and identified by (GTG)5-PCR fingerprinting, followed by gene sequencing of representative cluster isolates. Carnobacterium divergens, Carnobacterium maltaromaticum, Rahnella aquatilis, and Serratia proteamaculans were the most commonly found species, besides minor contributions of Lactobacillus sakei and Hafnia alvei. Differences in volatile profiles could thus be ascribed to variations in bacterial loads and storage temperatures.

Mapping the dominant microbial species diversity at expiration date of raw meat and processed meats from equine origin, an underexplored meat ecosystem, in the Belgian retail

Although equine meats and their derived smoked or fermented products are popular in some regions of the world, they only form a minor fraction of the global meat consumption. The latter may explain why their associated bacterial communities have not received much attention. In the present study, 69 different samples of equine meats and meat products were investigated. The samples consisted of raw meat from horses (17 samples) and zebra (7), as well as non-fermented but smoked (24) and fermented (21) horse meat products. After purchase, all samples were stored at 4 °C and analysed at expiration date. Besides an estimation of the total microbial counts, specific attention was paid to the identification of lactic acid bacteria (LAB) and catalase-positive cocci, in particular the group of coagulase-negative staphylococci (CNS), involved, due to their technological relevance in view of the elaboration of meat products. Samples that were loosely wrapped in butcher paper instead of vacuum- or modified-atmosphere packages were also screened for pseudomonads and enterobacterial species. In total, 1567 bacterial isolates were collected, subjected to (GTG)5-PCR fingerprinting of genomic DNA, and identified by multiple gene sequencing (based on the 16S rRNA, pheS, rpoA, rpoB, and/or tuf genes). Overall, the bacterial species diversity consisted mostly of LAB but was contingent on the type of product. Raw meat was dominated by Carnobacterium divergens, Lactobacillus sakei, Lactococcus piscium, and Leuconostoc gelidum, with zebra meat being particularly rich in lactococci. Smoked and fermented horse meat products contained mostly Lb. sakei and, to a lesser degree, Lactobacillus curvatus. In addition, several catalase-positive cocci (mostly Staphylococcus equorum), Anoxybacillus sp., Brevibacterium sp., Brochothrix thermosphacta, and the enterobacterial species Hafnia alvei were found.

Isolation and Characterization of Lactic Acid Bacteria in Philippine Fermented Milkfish Chanos chanos-Rice Mixture (Burong Bangus)

Isolation and characterization of bacteria in food products are important to determine and distinguish the beneficial or harmful effects of microbiota in certain samples. Lactic acid bacteria in food products had long been associated to good factors as food preservatives and with added fermentation metabolites. This study isolated and characterized lactic acid bacteria from burong bangus. The culture and purification process of bacteria isolation resulted to 4 strains of lactic acid bacteria namely Enterococcus faecalis, Tetragenococcus muriaticus, Lactobacillus delbrueckii subp. delbrueckii and Carnobacterium divergens. High enzymatic activity were observed with E. faecalis particularly on lipase and protease assay. While C. divergens have no enzymatic activity against lipase, protease, amylase and cellulase. The antimicrobial property of L. delbrueckii is only susceptible to amoxicillin unlike the other three bacteria isolates. No antagonistic activity were observed with the four bacterial strains against Bacillus subtilis, Staphylococcus aureus and Escherichia coli. The result of this study showed promising benefits to the industry especially in developing countries like the Philippines because population are not yet so aware of this organisms and the benefits that can be derived through their consumption.

Tracking the sources of psychrotrophic bacteria contaminating chicken cuts during processing

The major aim of the study was to establish the routes via which spoilage associated psychrotrophic bacteria contaminate poultry products at a large processing plant located in Belgium. Environmental samples were collected consisting of samples of air and swabs of food contact surfaces. Product samples were also collected consisting of modified atmosphere packaged (MAP) chicken wings and legs, which were analyzed microbiologically on the same day they were produced as well as after their sell-by date. Psychrotrophic bacteria from these samples were subsequently clustered and identified by means of MALDI-TOF MS and 16S rRNA gene sequencing. Carnobacterium maltaromaticum was determined to dominate the spoilage flora of both wings and legs. Other psychrotrophic bacteria able to grow on MRS which were identified on expired wings and legs included Carnobacterium divergens, Brocothrix thermosphacta, Lactobacillus curvatus, and Lactobacillus brevis. These were determined to arise from food contact surfaces such as cutting blades, leg hooks, Ertalon and polyurethane conveyor belts, working tables, and the hands of the operators. Importantly, it was determined that cleaning and disinfection was largely inadequate. Air was also determined to be an important vector of psychrotrophic bacteria in the processing environment, potentially contaminating the products directly or indirectly.

Assertiveness of meat-borne Lactococcus piscium strains and their potential for competitive exclusion of spoilage bacteria insitu and invitro.

This study aimed to investigate intraspecies assertiveness of meat-borne Lactococcus piscium isolates, inhibitory effects on unwanted and harmful meat spoilers, and the prevalence on beef deliberately inoculated with Lc. piscium. Co-inoculation of Lc. piscium isolates and spoilers (Brochothrix thermosphacta, Leuconostoc gelidum subsp. gasicomitatum, Carnobacterium divergens, Pseudomonas weihenstephanensis, Serratia liquefaciens, Hafnia alvei) were conducted in sterile meat simulation medium. Differentiation of Lc. piscium strains was carried out with colony-based RAPD-PCR. Selective cultivation was used to differentiate spoilers from Lc. piscium. Intraspecies assertiveness revealed Lc. piscium TMW2.1614 as most assertive strain. Co-inoculation of selected Lc. piscium strains caused substantial growth reduction of spoilers while the extent was strain- and spoiler dependent. Monitoring the microbiota on beef steaks deliberately inoculated with Lc. piscium revealed prevalence over the endogenous microbiota while maintaining a ripened sensory impression without undesired alterations. This study reveals Lc. piscium strains TMW2.1612/2.1614/2.1615 as highly competitive against spoilers invitro while beef deliberately inoculated with these strains maintained acceptable organoleptics. Selected Lc. piscium strains exhibit high potential for application as bioprotective cultures for competitive exclusion on beef in order to extend minimum shelf life and enhance product safety of meat.

Variability within the dominant microbiota of sliced cooked poultry products at expiration date in the Belgian retail

Sliced cooked poultry products are susceptible to bacterial spoilage, notwithstanding their storage under modified-atmosphere packaging (MAP) in the cold chain. Although the prevailing bacterial communities are known to be mostly consisting of lactic acid bacteria (LAB), more information is needed about the potential variation in species diversity within national markets. In the present study, a total of 42 different samples of sliced cooked poultry products were collected in the Belgian retail and their bacterial communities were analysed at expiration date. A total of 629 isolates from four different culture media, including plate count agar for the total microbiota and de Man-Rogosa-Sharpe (MRS), modified MRS, and M17 agar as three selective agar media for LAB, were subjected to (GTG)5-PCR fingerprinting and identification by gene sequencing. Overall, Carnobacterium, Lactobacillus, and Leuconostoc were the dominant genera. Within each genus, the most encountered isolates were Carnobacterium divergens, Lactobacillus sakei, and Leuconostoc carnosum. When comparing samples from chicken origin with samples from turkey-derived products, a higher dominance of Carnobacteria spp. was found in the latter group. Also, an association between the dominance of lactobacilli and the presence of added plant material and lactate salts was found.

Phenotypic and Genotypic Characterization of Carnobacterium divergens Isolated from Refrigerated Tunisian Minced Raw Beef Meat

Six psychotropic strains of Carnobacterium divergens were isolated from Tunisian minced raw beef meat packed and stored at 6°C. They were first identified by biochemical methods. Using biochemical reactions and carbohydrate fermentation before their characterisation by molecular techniques. The strain of Carnobacterium divergens is a nonmotile, Gram-positive psychotropic rod that lacks catalase, oxidase and mannitol. It grows at pH 9.1 (D-MRS agar), but not on acetate agar (pH ≤ 5.4). For all these isolates, the phenotypic identification, using the API 50 CHL system, revealed a variability in the fermentation abilities of some sugars (glycerol, amygdaline, arbutine, D-trehalose and potassium gluconate). Species-specific polymerase chain reaction (PCR) primers were used to ensure identification of these isolated strains at the species level. Moreover, the sequencing of 16S rRNA gene confirmed that all the six isolates are identified as C. divergens. The Rep-PCR technique was performed to investigate intra-specific diversity within these six strains of C. divergens, commonly identified in meat. The Rep-PCR method distinguished the C. divergens strains, demonstrating their biodiversity and their evident similarities. BOX and REP primers allowed amplification with 85% and 80% similarities between the strains, respectively. The BOXA1R and REP primers were proven to be useful for the differentiation of C. divergens strains.

Inhibition of Listeria Monocytogenes and Spoilage Bacteria on Cured Ready-to-Eat Meats by Sodium-Free and Clean-Label Antimicrobial Ingredients

ObjectivesTo compare the inhibitory properties of a standard potassium acetate and diacetate blend (Provian K) and a natural fermented and neutralized dry vinegar product (Provian NDV)on the growth of both Listeria monocytogenes and lactic acid bacteria in cured RTE meats.Materials and MethodsFive treatments of cured deli-style ham (72 to 74% moisture, 1.75 ± 0.1% salt, and pH 6.2 to 6.4, 156 mg/kg sodium nitrite and 547 mg/kg sodium erythorbate)where tested, including a control without antimicrobials and different concentrations of Provian K (0.5 and 0.75%) and Provian NDV (0.5%, 0.65%). Cooked products were surface-inoculated with 3-log10 CFU/g of a cocktail of 5 strains of Listeria monocytogenes (including serotypes 4b, 1/2a and 1/2b) or a mixture of 2 lactic acid bacteria (LAB) strains including Carnobacterium divergens and Leuconostoc mesenteroides, both isolated from spoiled cooked meat products. Inoculated slices (100 g/package) were vacuum-packaged and stored at different temperatures for up to 4 and 8 wk, for LAB and L. monocytogenes, respectively. Triplicate samples per treatment were assayed by enumerating twice on modified Oxford Agar for L. monocytogenes and on Plate Count agar (30°C, 48 h) and APT agar with bromocresol purple (27°C, 48 h) for LAB. The study was conducted twice.ResultsResults on Listeria inhibitionControl Ham supported > 1 log increase of L. monocytogenes at 4 and 2 wk storage at 4°C and 7°C, respectively (Fig. 1). In contrast, hams supplemented with 0.5 or 0.75% Provian K or 0.65% NDV inhibited the Listeria growth for 12 and 8 wk at 4 and 7°C, respectively. Inhibition of Listeria on ham supplemented with 0.5% NDV was further affected by pH and moisture. Ham supplemented with 0.5% Provian NDV in the trial 1 (71.5% moisture, pH 6.2) delayed Listeria for 12 wk storage at 4°C, whereas individual samples of trial 1 (72.9% moisture, pH 6.3) supported growth (> 1 log increase) at 8 wk. Similar trends were observed at 7°C.Results on Lactic Acid Bacteria inhibitionThe Control Ham supported the increase of Lactic acid bacteria to spoilage level (> 6 Logs) at 3 and 2 wk storage at 4°C and 7°C, respectively. In contrast, hams supplemented with 0.5% Provian K or 0.8% Provian NDV showed complete inhibition of LAB for 4 wk. For complete inhibition at 7°C slight higher concentrations of 0.75% and 1% were needed of Provian K and Provian NDV, respectively.ConclusionThe results of this study show the inhibitive effect of a natural fermented vinegar product on the growth of Listeria monocytogenes and Lactic acid bacteria in a cured ready-to eat ham. To have a comparable efficacy with neutralized natural vinegar as with a standard acetate-diacetate product, the dosage of the vinegar based product should be increased by approximately 10%. This can be explained by the lower amount of undissociated acid in the product.

Chicken fillets subjected to UV-C and pulsed UV light: Reduction of pathogenic and spoilage bacteria, and changes in sensory quality.

We have compared the efficacy of continuous ultraviolet (UV‐C) (254 nm) and pulsed UV light in reducing the viability of Salmonella Enteritidis, Listeria monocytogenes, Staphylococcus aureus, enterohemorrhagic Escherichia coli, Pseudomonas spp., Brochothrix thermospacta, Carnobacterium divergens, and extended‐spectrum β‐lactamase producing E. coli inoculated on chicken fillet surface. Fluences from 0.05 to 3.0 J/cm2 (10 mW/cm2, from 5 to 300 s) used for UV‐C light resulted in average reductions from 1.1 to 2.8 log cfu/cm2. For pulsed UV light, fluences from 1.25 to 18.0 J/cm2 gave average reductions from 0.9 to 3.0 log cfu/cm2. A small change in the odor characterized as sunburnt and increased concentration of volatile compounds associated with burnt odor posed restrictions on the upper limit of UV treatment, however no sensory changes were observed after cooking the meat. Treatments under modified atmosphere conditions using a UV permeable top film gave similar or slightly lower bacterial reductions.Practical applicationsUltraviolet (UV) light may be used for decontaminating the surface of food products and reduce viability of pathogenic and spoilage bacteria. Exposure of raw chicken fillet surface to various doses of continuous UV‐C or pulsed UV light proposed in the present work represent alternatives for microbiological improvement of this product. Chicken fillets can be treated in intact packages covered with UV permeable top film, thus avoiding recontamination of the meat. UV‐C light treatment is a low cost strategy with low maintenance, whereas pulsed UV light involves more elaborate equipment, but treatment times are short and less space is required. Both methods can be helpful for producers to manage the safety and quality of chicken fillets.

Purification of leucocin A for use on wieners to inhibit Listeria monocytogenes in the presence of spoilage organisms

The aims of this study were to improve the method for purification of leucocin A to increase yield of peptide and to evaluate the efficacy of leucocin A and an analogue of leucocin A (leucocin N17L) to inhibit the growth of Listeria monocytogenes on wieners in the presence of spoilage organisms. Leucocin A was produced by Leuconostoc gelidum UAL187 and purified with a five-fold increase in yield; leucocin N17L was synthesized replacing asparagine at residue 17 with leucine. Five strains of L. monocytogenes associated with foodborne illness were used to assess bacteriocin efficacy in vitro and in situ. Minimum inhibitory concentrations could not be determined in broth; however, on agar the minimum inhibitory concentrations ranged from 11.7–62.5μM and 62.5–>500μM for leucocin A and leucocin N17L, respectively. Leucocin N17L was less effective than the native bacteriocin at controlling the growth of L. monocytogenes. The inactivation profiles of L. monocytogenes in broth in the presence of leucocin A suggested each isolate had different levels of resistance to the bacteriocin as determined by the initial bactericidal effect. The formation of spontaneously resistance subpopulations were also observed for each strain of L. monocytogenes. In situ, wieners were inoculated with the spoilage organisms, Carnobacterium divergens and Brochothrix thermosphacta, followed by surface application of purified leucocin A, and inoculated with a cocktail of L. monocytogenes. Wieners were vacuum packaged and stored at 7°C for 16d. Leucocin A reduced the counts L. monocytogenes on wieners during storage, regardless of the presence of C. divergens. B. thermosphacta was unaffected by the presence of leucocin A on wieners over the duration of storage. This study suggests that leucocin A may be beneficial to industry as a surface application on wieners to help reduce L. monocytogenes counts due to post-processing contamination even in the presence of spoilage organisms. However, further investigation on the ability of L. monocytogenes to form spontaneous resistance to class II bacteriocins on food matrices during prolonged storage is warranted.

Scientific Opinion on the update of the list of QPS-recommended biological agents intentionally added to food or feed as notified to EFSA.

EFSA is requested to assess the safety of a broad range of biological agents in the context of notification for market authorisation as sources of food and feed additives, food enzymes and plant protection products. The qualified presumption of safety (QPS) assessment was developed to provide a harmonised generic pre‐assessment to support safety risk assessments performed by EFSA's scientific Panels. The safety of unambiguously defined biological agents (at the highest taxonomic unit appropriate for the purpose for which an application is intended), and the completeness of the body of knowledge are assessed. Identified safety concerns for a taxonomic unit are, where possible and reasonable in number, reflected as ‘qualifications’ in connection with a recommendation for a QPS status. The list of QPS recommended biological agents was reviewed and updated in the current opinion and therefore becomes the valid list. The 2016 update reviews previously assessed microorganisms including bacteria, yeasts and viruses used for plant protection purposes following an Extensive Literature Search strategy. The taxonomic units related to the new notifications received since the 2013 QPS opinion, were periodically evaluated for a QPS status and the results published as Statements of the BIOHAZ Panel. Carnobacterium divergens, Lactobacillus diolivorans, Microbacterium imperiale, Pasteuria nishizawae, Pediococcus parvulus, Bacillus flexus, Bacillus smithii, Xanthomonas campestris and Candida cylindracea were recommended for the QPS list. All taxonomic units previously recommended for the 2013 QPS list had their status reconfirmed as well their qualifications with the exception of Pasteuria nishizawae for which the qualification was removed. The exclusion of filamentous fungi and enterococci from the QPS evaluations was reconsidered but monitoring will be maintained and the status will be re‐evaluated in the next QPS Opinion update. Evaluation of bacteriophages should remain as a case‐by‐case procedure and should not be considered for QPS status.

Improving simultaneously the quality and safety of cooked and peeled shrimp using a cocktail of bioprotective lactic acid bacteria

Tropical shrimp is of considerable economic importance in the world but is highly perishable due to microbial and chemical degradation. Biopreservation is a food preservation technology based on the addition of “positive” bacteria able to kill or prevent the growth of undesirable microorganisms. Two strains of lactic acid bacteria (LAB) have previously been selected for a biopreservation strategy: Lactococcus piscium CNCM I-4031, for its ability to prevent the sensory deterioration of seafood and Carnobacterium divergens V41, which inhibits growth of Listeria monocytogenes. The objective was to test the association of the two strains to improve both the quality and safety of shrimp. In a first trial, the two LAB were inoculated alone or in a cocktail in cooked and peeled shrimp (CPS) Penaeus vannamei at 5×105CFU/g. Chemical, sensory and microbiological analyses by culture-dependent and -independent methods were performed during storage under modified atmosphere packaging (MAP) at 8°C. The results were compared to a non-inoculated batch. In a second trial, the same experiments were repeated in the presence of 102CFU/g of L. monocytogenes RF191. The microbiota of CPS was composed of LAB, Shewanella spp. and Enterobacteriaceae. Brochothrix thermosphacta was not detected. L. piscium and C. divergens reached 108 and 109CFU/g, respectively, in 7days and did not inhibit each other when co-inoculated. L. piscium reduced L. monocytogenes by 1Log (CFU/g) for 28days. C. divergens had an immediate listericidal effect lasting 7days. A regrowth of L. monocytogenes was then observed but the count was always 2 to 5Log (CFU/g) lower than in the control. No additional or synergic effect between protective strains was observed and the cocktail had the same inhibitory effect as C. divergens alone. C. divergens was very effective at preventing the sensory deterioration of CPS. This may be related to the inhibition of Shewanella and Enterobacteriaceae. However, the panelists could detect the presence of C. divergens during the first 10days of storage, with slight unpleasant odors and flavors. L. piscium improved the sensory quality of CPS for 14days only. In co-culture, L. piscium eliminated the off-odors and flavors released by C. divergens in the early stage of storage and the co-culture allowed maintaining a good quality of CPS throughout the storage. Therefore, the use of a cocktail of L. piscium CNCM I-4031 and C. divergens V41 is recommended in a strategy of biopreservation of shrimp.

Optimization of Growth and Bacteriocin Activity of the Food Bioprotective Carnobacterium divergens V41 in an Animal Origin Protein Free Medium

Optimization of Carnobacterium divergens V41 growth and bacteriocin activity in a culture medium deprived of animal protein, needs for food bioprotection, was performed by using a statistical approach. In a screening experiment, twelve factors (pH, temperature, carbohydrates, NaCl, yeast extract, soy peptone, sodium acetate, ammonium citrate, magnesium sulphate, manganese sulphate, ascorbic acid and thiamine) were tested for their influence on the maximal growth and bacteriocin activity using a two-level incomplete factorial design with 192 experiments performed in microtiter plate wells. Based on results, a basic medium was developed and three variables (pH, temperature and carbohydrates concentration) were selected for a scale-up study in bioreactor. A 23 complete factorial design was performed, allowing the estimation of linear effects of factors and all the first order interactions. The best conditions for the cell production were obtained with a temperature of 15°C and a carbohydrates concentration of 20 g/l whatever the pH (in the range 6.5-8), and the best conditions for bacteriocin activity were obtained at 15°C and pH 6.5 whatever the carbohydrates concentration (in the range 2-20 g/l). The predicted final count of C. divergens V41 and the bacteriocin activity under the optimized conditions (15°C, pH 6.5, 20 g/l carbohydrates) were 2.4 x 1010 CFU/ml and 819200 AU/ml respectively. C. divergens V41 cells cultivated in the optimized conditions were able to grow in cold-smoked salmon and totally inhibited the growth of Listeria monocytogenes (< 50 CFU g-1) during five weeks of vacuum storage at 4° and 8°C.

The effects of inulin, dried Jerusalem artichoke tuber and a multispecies probiotic preparation on microbiota ecology and immune status of the large intestine in young pigs

ABSTRACTThe study aimed at determining the effect of two types of prebiotics and a multispecies probiotic on microbiota activity and composition, as well as mucosal immunity in the large intestine of young pigs. In total 48 piglets were divided into 6 groups (n = 8), which received from day 10 of life probiotic-unsupplemented (PU) or probiotic-supplemented (PS) diets. Probiotics were added at 0.5 g/kg diet and contained: Lactococcus lactis, Carnobacterium divergens, Lactobacillus casei, Lactobacillus plantarum and Saccharomyces cerevisiae. The PU and PS diets were formulated without prebiotic addition (control) or with addition of 2% of inulin from chicory root (IN) or 4% of dried Jerusalem artichoke tubers (DJA). After 40 days of feeding, digesta and tissue samples were taken from the caecum and three sections of the colon for analyses of microbiota activity and composition, secretory immunoglobulin A (sIgA) and intraepithelial lymphocytes (IEL). IN diets decreased the caecal digesta pH and β-glucosidase activity but increased propionic, valeric and total short chain fatty acid (SCFA) concentrations compared to control diets. Feeding DJA diets increased caecal valeric acid level, decreased the concentration of isoacids in the colon, reduced β-glucosidase and β-glucuronidase activity in the middle colon and increased Bifidobacterium spp. populations in the proximal and distal colon. PS diets increased the caecal acetic acid and total SCFA level, and Clostridium spp. populations in the distal colon. Neither probiotic nor prebiotics affected sIgA level or IEL number in the large intestine. In conclusion, DJA modified the microbiota ecology in the large intestine of young pigs to a greater extent than IN and the applied probiotic did not enhance effects of prebiotics.

Reducing Salt in Raw Pork Sausages Increases Spoilage and Correlates with Reduced Bacterial Diversity.

Raw sausages are perishable foodstuffs; reducing their salt content raises questions about a possible increased spoilage of these products. In this study, we evaluated the influence of salt reduction (from 2.0% to 1.5% [wt/wt]), in combination with two types of packaging (modified atmosphere [50% mix of CO2-N2] and vacuum packaging), on the onset of spoilage and on the diversity of spoilage-associated bacteria. After 21 days of storage at 8°C, spoilage was easily observed, characterized by noticeable graying of the products and the production of gas and off-odors defined as rancid, sulfurous, or sour. At least one of these types of spoilage occurred in each sample, and the global spoilage intensity was more pronounced in samples stored under modified atmosphere than under vacuum packaging and in samples with the lower salt content. Metagenetic 16S rRNA pyrosequencing revealed that vacuum-packaged samples contained a higher total bacterial richness (n = 69 operational taxonomic units [OTUs]) than samples under the other packaging condition (n = 46 OTUs). The core community was composed of 6 OTUs (Lactobacillus sakei, Lactococcus piscium, Carnobacterium divergens, Carnobacterium maltaromaticum, Serratia proteamaculans, and Brochothrix thermosphacta), whereas 13 OTUs taxonomically assigned to the Enterobacteriaceae, Enterococcaceae, and Leuconostocaceae families comprised a less-abundant subpopulation. This subdominant community was significantly more abundant when 2.0% salt and vacuum packaging were used, and this correlated with a lower degree of spoilage. Our results demonstrate that salt reduction, particularly when it is combined with CO2-enriched packaging, promotes faster spoilage of raw sausages by lowering the overall bacterial diversity (both richness and evenness). Our study takes place in the context of raw meat product manufacturing and is linked to a requirement for salt reduction. Health guidelines are calling for a reduction in dietary salt intake. However, salt has been used for a very long time as a hurdle technology, and salt reduction in meat products raises the question of spoilage and waste of food. The study was conceived to assess the role of sodium chloride reduction in meat products, both at the level of spoilage development and at the level of bacterial diversity, using 16S rRNA amplicon sequencing and raw pork sausage as a meat model.

Selection of bioprotective cultures for preventing cold-smoked salmon spoilage

Biopreservation is a natural technology of food preservation, which consists of inoculating food with microorganisms selected for their antibacterial properties. The objective of this study was to select lactic acid bacteria (LAB) to improve the quality of cold-smoked salmon (CSS). In this work, different strains representative of the 4 dominant species, identified in a previous study by pyrosequencing the 16S rRNA gene, were isolated and their spoiling potential in CSS blocks, sterilized by ionization, was assessed by twelve trained panelists along the vacuum storage at 8°C. Photobacterium phosphoreum, Brochothrix thermosphacta and Serratia proteamaculans released strong off-odors whereas the spoiling potential of Carnobacterium divergens was weaker. The spoiling capacity of Lactococcus piscium EU2241, Leuconostoc gelidum EU2247, Lactobacillus sakei EU2885, Staphylococcus equorum S030674 and 4 commercial starters was tested by the same method and 2 strains were eliminated due to off-odor production. The effect of the 6 selected LAB against the 4 specific spoiling organisms (SSOs) selected was tested by challenge tests in sterile CSS blocks. The protective effect of the LAB differed from one SSO to another and no correlation could be established between the sensory improvement, SSO inhibition, and the implantation or acidification of protective cultures (PCs). All the PCs except L. piscium reduced the off-odors released by P. phosphoreum although some of them had no effect on its growth. S. equorum, which did not grow in CSS, favored the implantation of P. phosphoreum but prevented its off-odor formation. L. piscium was the only strain that prevented the spoilage of B. thermosphacta and S. proteamaculans although it did not grow very well and did not acidify the product. L. gelidum EU2247 inhibited the growth of these 2 SSOs and lowered the pH but had no effect on the sensory quality. Finally, L. piscium was tested in 2 naturally contaminated products, with a positive effect on 1 batch. This effect was not correlated with the microbial ecosystem as determined by acultural and cultural techniques. Based on these results, the selection strategy is discussed.

Carnobacterium Isolated from Caviar of Sturgeon (Acipenser ruthenus) Farmed in Korea

Sturgeon aqua-cultured in Korea is mainly Acipenser ruthenus and its culture began in the early 2000s. In this study, Carnobacterium sp. was isolated from unprocessed caviar of aqua-cultured Acipenser ruthenus. The 16s rRNA nucleotide sequence obtained from Carnobacterium sp. isolate (accession no. KM236206) was deposited with GenBank and homologous with Carnobacterium divergens DSM 20623 and NBRC15683 strain. In conclusion, this is first report of isolation of Carnobacterium sp. from caviar of Acipenser ruthenus aqua-cultured in Korea. In the future, it must be ascertained whether Carnobacterium sp. degenerate of caviar or cause diseases in sturgeon.

Microbiological changes, shelf life and identification of initial and spoilage microbiota of sea bream fillets stored under various conditions using 16S rRNA gene analysis.

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. 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. 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.

Prolonged Shelf Life and Reduced Drip Loss of Chicken Filets by the Use of Carbon Dioxide Emitters and Modified Atmosphere Packaging

Modified atmosphere packaging containing CO2 is widely used for extending the shelf life of chicken meat. Active packaging by adding CO2 emitter sachets to packages of meat is an alternative to traditional modified atmosphere packaging. The purpose of the study was to investigate the shelf life of chicken filets under different CO2 concentrations at 4°C storage. The inhibition of microbial growth was proportional to the CO2 concentration. Storage in 100% CO2 both with and without a CO2 emitter sachet gave a microbiological shelf-life extension of 7 days compared with 60% CO2. Carnobacterium divergens, Carnobacterium sp., and Lactococcus sp. were the dominating species at the end of the storage period. During storage in pure CO2, the carbon dioxide dissolved in the meat and caused the collapse of the packages. The resulting squeeze of the meat lead to a severe increase in drip loss. The drip loss was reduced profoundly by using the CO2 emitting sachet in the packages. The addition of CO2 emitters can easily be implemented at industrial packaging lines without reduction in production efficiency.

Microbial successions are associated with changes in chemical profiles of a model refrigerated fresh pork sausage during an 80-day shelf life study.

Fresh pork sausage is produced without a microbial kill step and therefore chilled or frozen to control microbial growth. In this report, the microbiota in a chilled fresh pork sausage model produced with or without an antimicrobial combination of sodium lactate and sodium diacetate was studied using a combination of traditional microbiological methods and deep pyrosequencing of 16S rRNA gene amplicons. In the untreated system, microbial populations rose from 10(2) to 10(6) CFU/g within 15 days of storage at 4°C, peaking at nearly 10(8) CFU/g by day 30. Pyrosequencing revealed a complex community at day 0, with taxa belonging to the Bacilli, Gammaproteobacteria, Betaproteobacteria, Actinobacteria, Bacteroidetes, and Clostridia. During storage at 4°C, the untreated system displayed a complex succession, with species of Weissella and Leuconostoc that dominate the product at day 0 being displaced by species of Pseudomonas (P. lini and P. psychrophila) within 15 days. By day 30, a second wave of taxa (Lactobacillus graminis, Carnobacterium divergens, Buttiauxella brennerae, Yersinia mollaretti, and a taxon of Serratia) dominated the population, and this succession coincided with significant chemical changes in the matrix. Treatment with lactate-diacetate altered the dynamics dramatically, yielding a monophasic growth curve of a single species of Lactobacillus (L. graminis), followed by a uniform selective die-off of the majority of species in the population. Of the six species of Lactobacillus that were routinely detected, L. graminis became the dominant member in all samples, and its origins were traced to the spice blend used in the formulation.