Growth of Carnobacterium divergens V41 and production of biogenic amines and divercin V41 in sterile cold-smoked salmon extract at varying temperatures, NaCl levels, and glucose concentrations

A complete factorial design 2(3) was used to study some aspects of Carnobacterium divergens V41 metabolism (growth, biogenic amine production, and divercin V41 production) in sterile cold-smoked salmon extract (SSE) at varying temperatures (3 to 9 degrees C), NaCl levels (2.5 to 6.5%), and glucose concentrations (2 to 6 g liter(-1)). The results showed that temperature and NaCl content were the most influential factors on growth parameters in SSE. Predictive models are suggested for the assessment of C. divergens lag time (t(lag)) and maximum specific growth rate (micro(max)) Among the biogenic amines studied, only tyramine was found to be produced by C. divergens in SSE. Furthermore, we showed that temperature, NaCl, and glucose variations did not greatly affect tyramine and divercin V41 production by the bacteria under the experimental conditions used. Indeed, divercin V41, a bacteriocin from C. divergens V41 that is highly active against some Listeria strains, was produced in SSE even under harsh culture conditions. Similarly, tyramine production in SSE was delayed at 3 degrees C but reached 35 microg ml(-1) in all experiments after 27 days of storage. However, this final tyramine concentration in SSE is low compared with the threshold values of 100 to 800 microg g(-1) reported as the potentially toxic dose in foods. Thus, we have found that C. divergens V41 is a promising strain for the biopreservation of refrigerated cold-smoked salmon.     

Microbial selenate sorption and reduction in nutrient limited systems

In this study, batch sorption experiments and X-ray adsorption spectroscopy (XAS) were utilized to investigate selenate sorption onto Shewanella putrefaciens 200R. Selenate sorption was studied as a function of pH (ranging from 3 to 7), ionic strength (ranging from 0.1 to 0.001 M), and initial selenate concentration (ranging from 10 to 5000 microM) in the absence of external electron donors. The results show that the extent of selenate sorption is strongly dependent on pH and ionic strength, with maximum sorption occurring at low pH (pH = 3) and low ionic strength (0.001 M NaCl) conditions. The strong dependence of Se sorption with ionic strength suggests the formation of outersphere complexes with the cell wall functional groups. Langmuir isotherm plots yielded log Kads values from 2.74 to 3.02. Desorption experiments demonstrated thatthe binding of selenate onto S. putrefaciens was not completely reversible. XANES analysis of the cells after sorption experiments revealed the presence of elemental selenium, indicating that S. putrefaciens has a capacity to reduce Se(VI) to Se(0) in the absence of external electron donors. We conclude that Se sorption onto S. putrefaciens cell walls is the result of the combination of outer-sphere complexation and cell surface reduction. This sorption process leads to a complex reservoir of bound Se which is not entirely reversible.     

New procedure for the study of odour representativeness of aromatic extracts from smoked salmon

The representativeness of an aromatic extract of smoked salmon obtained from simultaneous steam distillation and extraction with diethyl ether is discussed. After extraction, the extract is diluted in ethanol with an evaporation of diethyl ether, which allows the extract to be redeposited on matrices physically similar to those of the original product. When the shift of the matrix effect is taken into account, the sensorial results are closer to reality and more representative of the real interaction conditions between the matrix and the extract. Several sensory methods are used to describe the representativeness of the smoked salmon extracts, such as triangular and notation tests. Preliminary work is carried out on standards known both to contribute to the aroma of many seafood products and to have a high volatility in comparison with those expected in smoked salmon in order to test the method in difficult conditions of recovery. This enables the recovery yield of the extraction (from 62% for limonene to 97% for 2-methylphenol) to be assessed leading to a better characterization of the representativeness taking into account the odour dilutions. The final aromatic extract is found to be about 70% representative of the original product.     

底物和环境因子对鱼源腐败希瓦氏菌和假单胞菌生长动力学的影响

水产品在贮运和加工过程中极易腐败,微生物是引起腐败的主要原因,腐败希瓦氏菌和假单胞菌分别是低温有氧贮藏海水鱼和淡水鱼时的特定腐败菌。为探讨环境因子和底物对特定腐败菌的影响,以源自大黄鱼和罗非鱼中的腐败希瓦氏菌和假单胞菌为对象,探究不同底物、温度、pH和NaCl影响下的菌体生长动态,采用Gompertz方程拟合生长曲线,定量分析对其最大比生长速率(μ_max)和迟滞期(λ)的影响,并使用Belehradek方程描述其μ_max与温度间的关系。结果表明:底物对腐败希瓦氏菌和假单胞菌生长影响较为明显;而温度对其生长影响最为显著,是影响腐败速率的决定性因素;pH对腐败希瓦氏菌生长影响较大,pH为7时其μ_max高于pH为6时,但对假单胞菌无明显影响,pH为4和5时,两者均不生长;NaCl含量对腐败希瓦氏菌生长有显著影响,随着NaCl含量的升高,其μ_max和λ降低。该研究可为探究水产品腐败菌调控因子筛选、靶向抑菌和延长产品货架期等提供支撑。     

Sodium and potassium transport in the halophilic yeast Debaryomyces hansenii

Debaryomyces hansenii, a halophile yeast found in shallow sea waters and salty food products grows optimally in 0.6 M of either NaCl or KCl, accumulating high concentrations of Na(+) or K(+). After growth in NaCl or KCl, a rapid efflux of either accumulated cation was observed if the cells were incubated in the presence of KCl or NaCl, respectively, accompanied by a slower accumulation of the cation present in the incubation medium. However, a similar, rapid efflux was observed if cells were incubated in buffer, in the absence of external cations. This yeast shows a cation uptake activity of both (86)Rb(+) and (22)Na(+) with saturation kinetics, and much higher affinity for (86)Rb(+) than for (22)Na(+). The pH dependence of the kinetics constants was similar for both cations, and although K(m) values were higher at pH 8.0, there was also an increase in the V(max) values. The accumulation of (22)Na(+) was found to be increased in cells grown in the presence of 0.6 M NaCl. (86)Rb(+) was also accumulated more in these cells, but to a slightly greater extent. The inhibition kinetics of the uptake of (22)Na(+) by K(+), and that of (86)Rb(+) by Na(+) was found to be non-competitive. It can be concluded that Na(+) in D. hansenii is not excluded but instead, its metabolic systems must be resistant to high salt concentrations.     

Biological Characteristics Affect the Quality of Farmed Atlantic Salmon and Smoked Muscle

Various biological characteristics influencing the quality of farmed salmon and smoked muscle were studied. No great differences in proximate composition were observed among raw fish. Stress produced a slight decrease in protein solubility at 0.8 M NaCl and also slight variations in electrophoretic profile. This was accompanied by a certain degree of muscle softening. Thirty-days starvation produced slight depletion of the sarcoplasmic fraction, collagen insolubilization, and muscle hardening. The effect of triploidy was more evident in sea-caged fish, resulting in lower protein solubility at 0.05M NaCl and lower insoluble collagen than diploids. After smoking, protein solubility at 0.8M NaCl was highest in stressed fish, and non-starved fish collagen became insolubilized.     

Use of sodium nitrite in salt-curing of Atlantic salmon (Salmo salar L.) – Impact on product quality

Quality changes during processing and quality differences in smoked fillets of Atlantic salmon (4–5 kg) salted with nitrite salt compared to table salted fillets were measured. Quality parameters from right-side fillets dry salted with nitrite salt were compared with the respective left-side fillets treated the same way with table salt. Ten raw right-side fillets were analysed and used as raw material reference. Use of nitrite salt in salt-curing of smoked salmon affected colour to a more reddish hue, tended to increase carotenoid stability and displayed positive effects on NaCl diffusivity. Only slight weight changes and change in texture properties were revealed. The use of nitrite salt displayed no adverse effects like increased content of N-nitrosoamines in smoked products. In fact, significant lower contents of N-nitrosoamines were found in nitrite salted smoked fillets compared to smoked fillets salted with table salt. Relatively high amounts of residual nitrite in nitrite-treated fillets seem to be the most prominent adverse effect caused by the use of nitrite salt in salt-curing of smoked Atlantic salmon.     

COMBINATION WITH PLANT EXTRACTS IMPROVES THE INHIBITORY ACTION OF DIVERGICIN M35 AGAINST LISTERIA MONOCYTOGENES

ABSTRACT

The susceptibility of 11 strains of Listeria monocytogenes to divergicin M35, a bacteriocin produced by Carnobacterium divergens strain M35, and to aqueous extracts of garlic, onion, oregano, red chili and black pepper at 30 and 10C, was evaluated using a microdilution assay. The susceptibility of divergicin‐resistant strains to combinations of these agents was also evaluated. Three strains were resistant to divergicin M35 (>500 µg/mL) at 30C but were more susceptible at 10C. Garlic gave the most inhibitory plant extract, followed by onion, while oregano, red chili and black pepper extracts were less active at both temperatures. Garlic extract and divergicin M35 combined or with other extracts increased inhibitory activity against the divergicin‐resistant strains. The garlic/divergicin combination was the most effective at inhibiting these strains and was bactericidal at both temperatures. Log‐phase cells were the most susceptible to the garlic/divergicin combination. Stationary‐phase cells were much more resistant at both incubation temperatures. Furthermore, the effect of the garlic/divergicin combination at inhibiting divergicin‐resistant L. monocytogenes in a food system was also studied using cold‐smoked salmon as a food model. Results indicated that this combination could efficiently reduce the viability of L. monocytogenes in smoked salmon stored at 10C.

PRACTICAL APPLICATIONS

There is increasing popularity worldwide for chemical preservative‐free, ready‐to‐eat and minimally processed seafood with low salt, fat and sugar content. Bacteriocins produced from lactic acid bacteria can have a potential application to prolong the shelf life of cold‐smoked salmon. Also, plant and spice extracts have been shown to contain antibacterial substances with potential for application in foods. Thus, this research explores the combination of divergicin M35, a bacteriocin produced by Carnobacterium divergens strain M35, and aqueous extracts of garlic, onion, oregano, red chili and black pepper to inhibit Listeria monocytogenes and to prolong the shelf life of cold‐smoked salmon.     

New method for rapid and sensitive quantification of sulphide-producing bacteria in fish from arctic and temperate waters

The offensive, fishy, rotten H2S-off-odours in spoiled, aerobically and cold stored fish from arctic and temperate waters are generally caused by sulphide-producing bacteria (SPB), mainly Shewanella putrefaciens. In the present work, a new, rapid, simple and accurate method for estimation of the SPB content in fish from these areas is described. The quantification is based on the formation rate of iron sulphide during growth of SPBs incubated at 30 degrees C in a liquid growth medium containing cysteine, sodium thiosulphate and iron(III)citrate as specific substrates for iron sulphide formation. The iron sulphide turns the medium grey and masks the background fluorescence in the medium when the SPB content in the assay is approximately 10(9) cfu/ml. The fluorescence change could be detected instrumentally and the colour change visually. The method was developed and evaluated in tests with S. putrefaciens CCUG 13452 DT as well as naturally occurring SPBs in cod, salmon, wolf fish and coal fish. A linear correlation between the SPB count and detection time was obtained over the entire range from 1 to 10(9) cfu SPB/g, corresponding to detection times 17 and 1 h, respectively. The correlation is described by the equation: log cfu/g fish= -0.59(+/- 0.17) x DT+ 9.65(+/- 0.09), where DT is the detection time in hours. The model was valid for all the tested fish species and all tested naturally occurring SPBs in these species. The regression coefficients (R2) for cod, coal fish, wolf fish and salmon were 0.99, 0.92, 0.97 and 0.97, respectively. The detection level of the method is 1 SPB per sample tube, corresponding to 16 cfu/g fish. The method could be used to predict the remaining shelf life of the fish for different markets, even when the time-temperature history during storage of the fish is unknown.     

Effect of salt, smoke compound, and storage temperature on the growth of Listeria monocytogenes in simulated smoked salmon

Smoked salmon can be contaminated with Listeria monocytogenes. It is important to identify the factors that are capable of controlling the growth of L. monocytogenes in smoked salmon so that control measures can be developed. The objective of this study was to determine the effect of salt, a smoke compound, storage temperature, and their interactions on L. monocytogenes in simulated smoked salmon. A six-strain mixture of L. monocytogenes (10(2) to 10(3) CFU/g) was inoculated into minced, cooked salmon containing 0 to 10% NaCl and 0 to 0.4% liquid smoke (0 to 34 ppm of phenol), and the samples were stored at temperatures from 0 to 25 degrees C. Lag-phase duration (LPD; hour), growth rate (GR; log CFU per hour), and maximum population density (MPD; log CFU per gram) of L. monocytogenes in salmon, as affected by the concentrations of salt and phenol, storage temperature, and their interactions, were analyzed. Results showed that L. monocytogenes was able to grow in salmon containing the concentrations of salt and phenol commonly found in smoked salmon at the prevailing storage temperatures. The growth of L. monocytogenes was affected significantly (P < 0.05) by salt, phenol, storage temperature, and their interactions. As expected, higher concentrations of salt or lower storage temperatures extended the LPD and reduced the GR. Higher concentrations of phenol extended the LPD of L. monocytogenes, particularly at lower storage temperatures. However, its effect on reducing the GR of L. monocytogenes was observed only at higher salt concentrations (>6%) at refrigerated and mild abuse temperatures (< 10 degrees C). The MPD, which generally reached 7 to 8 log CFU/g in salmon that supported L. monocytogenes growth, was not affected by the salt, phenol, and storage temperature. Two models were developed to describe the LPD and GR of L. monocytogenes in salmon containing 0 to 8% salt, 0 to 34 ppm of phenol, and storage temperatures of 4 to 25 degrees C. The data and models obtained from this study would be useful for estimating the behavior of L. monocytogenes in smoked salmon.     

Characterisation of volatile compounds produced by bacteria isolated from the spoilage flora of cold-smoked salmon.

This study investigated the volatile compounds produced by bacteria belonging to nine different bacterial groups: Lactobacillus sake, L. farciminis, L. alimentarius, Carnobacterium piscicola, Aeromonas sp., Shewanella putrefaciens, Brochothrix thermosphacta, Photobacterium phosphoreum and Enterobacteriaceae isolated from cold-smoked salmon. Each bacterial group was represented by several strains. In addition, combinations of the groups were examined as well. Sterile blocks of cold-smoked salmon were inoculated, vacuum-packed and stored at 6 degrees C. After 40 days of storage at 6 degrees C, aerobic viable count and pH were recorded, the volatile fraction of the samples was analysed by gas chromatography-mass spectrometry (GC-MS), and spoilage was assessed by sensory evaluation. Among the 81 volatile compounds identified by GC-MS, 30 appeared to be released as a result of bacterial metabolism. Some of the effects of inoculated bacterial strains on the composition of the volatile fraction seemed to be characteristic of certain bacterial species. Sensory analysis showed relationships between bacteria, the composition of the volatile fraction and the organoleptic quality of smoked salmon.     

Control of Listeria monocytogenes and Salmonella anatum on the surface of smoked salmon coated with calcium alginate coating containing oyster lysozyme and nisin

ABSTRACT:  This study investigated the antimicrobial effect of oyster lysozyme with or without nisin added to calcium alginate (CaAlg) coated on the surface of smoked salmon against Listeria monocytogenes and Salmonella anatum . L. monocytogenes or S. anatum inoculated smoked salmon samples (1 g) were dipped into CaAlg with either oyster lysozyme (OysL) or hen egg white lysozyme (HEWL), with or without added nisin (N), then stored at 4 °C for 35 d. Our results indicated that the effectiveness of oyster lysozyme or hen egg white lysozyme was enhanced when added to calcium alginate coatings. After 35 d at 4 °C the growth of L. monocytogenes and S. anatum was suppressed in the range of 2.2 to 2.8 log CFU/g with CaAlgNOysL or CaAlgNHEWL coatings compared to the control nontreated samples. There was no significant difference between oyster lysozyme and hen egg white lysozyme treatments against L. monocytogenes or S. anatum inoculated on the surface of salmon. Calcium alginate coatings containing lysozyme with nisin or without could be used to reduce the growth of L. monocytogenes and S. anatum on the surface of ready-to-eat smoked salmon at refrigerated temperatures.     

Utilization of smoked salmon trim in extruded smoked salmon jerky

During smoked salmon processing, the dark meat along the lateral line is removed before packaging; this by-product currently has little economic value. In this study, the dark meat trim was incorporated into an extruded jerky. Three formulations were processed: 100% smoked trim, 75% : 25% smoked trim : fresh salmon fillet, and 50% : 50% smoked trim : fresh salmon blends (w/w basis). The base formulation contained salmon (approximately 83.5%), tapioca starch (8%), pregelatinized potato starch (3%), sucrose (4%), salt (1.5%), sodium nitrate (0.02%), and ascorbyl palmitate (0.02% of the lipid content). Blends were extruded in a laboratory-scale twin-screw extruder and then hot-smoked for 5 h. There were no significant differences among formulations in moisture, water activity, and pH. Protein was highest in the 50 : 50 blend jerky. Ash content was highest in the jerky made with 100% trim. Total lipids and salt were higher in the 100% trim jerky than in the 50 : 50 blend. Hot smoking did not adversely affect docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) content in lipids from 100% smoked trim jerky. Servings of salmon jerky made with 75% and 100% smoked trim provided at least 500 mg of EPA and DHA. The 50 : 50 formulation had the highest Intl. Commission on Illumination (CIE) L*, a*, and b* color values. Seventy consumers rated all sensory attributes as between "like slightly" and "like moderately." With some formulation and processing refinements, lateral line trim from smoked salmon processors has potential to be incorporated into acceptable, healthful snack products. PRACTICAL APPLICATION: Dark meat along the lateral line is typically discarded by smoked salmon processors. This omega-3 fatty acid rich by-product can be used to make a smoked salmon jerky that provides a convenient source of these healthful lipids for consumers.     

Random amplified polymorphic DNA typing applied to the study of cross-contamination by Listeria monocytogenes in processed food products

The presence of Listeria spp. was investigated in 369 samples of cooked meat products and 52 of smoked salmon. Incidences of 17.6% for cooked meat and 38.5% for smoked salmon samples were found. All Listeria monocytogenes isolates (34 from meat products and 16 from smoked salmon) were typed serologically and by random amplified polymorphic DNA (RAPD) typing using primers HLWL74 (5'-ACGTATCTGC-3'), HLWL85 (5'-ACAACTGCTC-3'), and OMP-01 (5'-GTI'GGTGGCT-3'). Strains from cooked meat products were characterized and compared in relation to their origin. The detection of identical strains in products of different type and brand packed on the same date suggested cross-contamination, probably during the slicing process. All L monocytogenes isolates from smoked salmon were indistinguishable by serotyping and RAPD, suggesting that this strain was highly disseminated and adapted to the treatment used for the preservation of this food. RAPD subtypes were analyzed using GelCompar version 4.1 software and the unweighted pair method using arithmetic averages, and six groups with at least 78% similarity were established. Serotyping and RAPD results were in concordance, although RAPD showed a higher discriminatory power with L. monocytogenes isolates from meat products. RAPD is an easy method that could be useful to detect cross-contamination occurring during postprocessing manipulations.     

Timed Emulsification Studies with Chicken Breast Muscle: Whole Muscle, Low-Salt Washed Muscle and Low-Salt Soluble Proteins

Emulsion formation with chicken breast muscle was investigated using timed emulsification. Emulsions were made at a fixed oil/water ratio of 2:1 by Omni-mixing for various lengths of time between 0 and 5 min; then the emulsions were centrifuged and the aqueous layer analyzed for protein. Emulsions formed using whole muscle or muscle washed with low molarity salt solution and suspended in buffered (10 mM sodium phosphate, pH 7.0) 0.6M NaCl were superior in stability upon centrifugation to those made with muscle in distilled water, buffered 0.025M NaCl or buffered 0.05M NaCl. Size of insoluble protein pellets from the centrifuged emulsions decreased as emulsification time increased. Little emulsifying effect of the low-salt soluble (sarcoplasmic) protein, defined as those proteins extracted with buffered 0.05M NaCl, was observed in the presence or absence of high-salt solubilized protein.     

Listeria monocytogenes in the smoked salmon industry

Smoked salmon is sporadically contaminated with Listerial monocytogenes. Contamination levels are normally low and consumers are probably seldom exposed to risk concentrations. No clones of L. monocytogenes seem to be specific to smoked salmon, some clones found in smoked salmon having been isolated from several sources, including patients. Cold-smoking has been shown to eliminate L. monocytogenes in challenge tests at temperatures from 17.1 to 21.1 degrees C, while from 22.2 to 30 degrees C the bacteria survived. Under natural cold-smoking conditions (19 to 22 degrees C) the frequency and level of L. monocytogenes seems to decrease. Hot-smoking seems to eliminate the bacteria when smoke is applied during the whole heating process. The prevention of recontamination of both cold-smoked and hot-smoked salmon is therefore of great importance. L. monocytogenes multiply considerably in smoked salmon during storage. Growth is faster in challenge tests than in naturally-contaminated smoked salmon. The declared shelf-life under refrigeration should be shorter than that customarily stipulated by many producers. While the sources of L. monocytogenes in smoked salmon processing plants have still to be determined, raw salmon does not seem to be an important source. The main issue for producers is to prevent colonization of the processing environment and spread of the bacteria to products. This should be achieved by the systemic implementation of hygienic measures, including the HACCP approach.     

Development of a defatted mustard meal-based composite film and its application to smoked salmon to retard lipid oxidation

An antioxidant composite film was developed from defatted mustard meal (DMM) without incorporating external antioxidants and its applicability as a coating to smoked salmon has been investigated. Film-forming variables included the concentration of xanthan, heat treatment, and high pressure homogenisation. Tensile strength, elongation, water vapour permeability, and water solubility of composite films of DMM and xanthan were 5.1–8.8 MPa, 2.9–5.0%, 1.5–2.4 g mm/kPa/h/m², and 1.1–1.6%, respectively. The composite film with xanthan at 5% (w/w) demonstrated antioxidant properties and the coating with the solution forming the composite film (5% xanthan) retarded lipid oxidation and significantly reduced volatile changes in smoked salmon during storage at 4, 10, and 20 °C for 21 days. Coated salmon was preferred to uncoated salmon in glossiness and fish smell. The composite coating improved the stability of smoked salmon against lipid oxidation without imparting a negative sensory quality to the salmon.     

The effect of osmo-induced stress on product formation by Zymomonas mobilis on sucrose

The intensification of biosynthesis of fructooligosaccharides in the presence of high salt concentrations was observed during sucrose (10%) fermentation by Zymomonas mobilis 113S. A 0.6 M NaCl concentration led to an increase of oligosaccharide productivity by 3.5-fold. Sorbitol formation was increased in the presence of 0.16 M NaCl and was inhibited at highest salt concentrations. In a medium with high (65%, w/w) sucrose content the salts gave inhibitory effects on fructooligosaccharide production by lyophilised Z. mobilis cells. Influence of salts on gluconic acid and sorbitol formation under these conditions was studied. The ratio of oligosaccharides and gluconic acid productivity (Qolig./Qglucon.) was increased approximately 2 times at 1% KCl. Sorbitol formation was not significantly influenced in the presence of KCl (up to 2%).     

Timed Emulsification Studies with Chicken Breast Muscle: Soluble and Insoluble Myofibrillar Proteins

Emulsion formation with chicken breast muscle was investigated using timed emulsification. High-salt soluble proteins (pH 7.0, 0.6M NaCl) extracted from previously washed muscle (pH 7.0, 0.05M NaCl) were removed from the aqueous phase as mixing time increased. Low- and high-salt exhaustively washed muscle, resuspended in either 0.15 or 0.6M NaCl, pH 7.0, still exhibited good emulsion properties. The pellet protein decreased (> 90%) as mixing time increased from 0 to 5 min. Addition of sodium pyrophosphate to 0.6M NaCl suspensions of 0.05M NaCl washed muscle resulted in an increase in solubility of myofibrillar proteins and a general improvement in emulsification properties. High-salt insoluble proteins may play an important role in emulsion formation.