Spoilage evaluation of raw Atlantic salmon (Salmo salar) stored under modified atmospheres by multivariate statistics and augmented ordinal regression.

The development of quality monitoring systems for perishable food products like seafood requires extensive data collection under specified packaging and storage conditions, followed by advanced data analysis and interpretation. Even though the benefits of using volatile organic compounds as food quality indices have been recognized, few studies have focused on real-time quantification of the seafood volatilome and subsequent systematic identification of the most important spoilage indicators. In this study, spoilage of Atlantic salmon (Salmo salar) stored under modified atmospheres (% CO2/O2/N2) and air was characterized by performing multivariate statistical analysis and augmented ordinal regression modelling for data collected by microbiological, chemical and sensory analyses. Out of 25 compounds quantified by selected-ion flow-tube mass spectrometry, ethanol, dimethyl sulfide and hydrogen sulfide were found characteristic under anaerobic conditions (0/0/100 and 60/0/40), whereas spoilage under air was primarily associated with the production of alcohols and ketones. Under high-O2 MAP (60/40/0), only 3-methylbutanal fulfilled the identification criteria. Overall, this manuscript presents a systematic and widely applicable methodology for the identification of most potential seafood spoilage indicators within the context of intelligent packaging technology development. In particular, parallel application of statistics and modelling was found highly beneficial for the performance of the quality characterization process and for the practical applicability of the obtained results in food quality monitoring.

Microbiological, chemical and sensory spoilage analysis of raw Atlantic cod (Gadus morhua) stored under modified atmospheres.

During fish spoilage, microbial metabolism leads to the production of volatile organic compounds (VOCs), characteristic off-odors and eventual consumer rejection. The aim of the present study was to contribute to the development of intelligent packaging technologies by identifying and quantifying VOCs that indicate spoilage of raw Atlantic cod (Gadus morhua) under atmospheres (%v/v CO2/O2/N2) 60/40/0, 60/5/35 and air. Spoilage was examined by microbiological, chemical and sensory analyses over storage time at 4 or 8 °C. Selected-ion flow-tube mass spectrometry (SIFT-MS) was used for quantifying selected VOCs and amplicon sequencing of the 16S rRNA gene was used for the characterization of the cod microbiota. OTUs classified within the Photobacterium genus increased in relative abundance over time under all storage conditions, suggesting that Photobacterium contributed to spoilage and VOC production. The onset of exponential VOC concentration increase and sensory rejection occurred at high total plate counts (7-7.5 log). Monitoring of early spoilage thus calls for sensitivity for low VOC concentrations.

Optimization and evaluation of a decontamination step with peroxyacetic acid for fresh-cut produce.

Since several disadvantages are associated with the use of sodium hypochlorite as a decontamination agent, the attention for alternative agents such as peroxyacetic acid (PAA) is increasing. In this study the effectiveness of PAA to remove the native microflora was tested in four types of fresh-cut vegetables: grated carrots, fresh-cut white cabbage, iceberg lettuce and leek. Furthermore, the influence of varying PAA concentrations (0, 25, 80, 150 and 250 ppm) and varying contact times (1, 5 and 10 min) was described by means of a linear model. The efficiency of PAA to remove the native flora was highly dependent on the type of fresh-cut produce: the highest microbial reductions were obtained for carrots (0.5-3.5 log cfu/g) and white cabbage (0.5-3.5 log cfu/g) followed by iceberg lettuce (0.4-2.4 log cfu/g). The obtained efficiency was the lowest for fresh-cut leek (0.4-1.4 log cfu/g). Furthermore, all the treated samples, regardless of the type of vegetable and the contact time and concentration of the PAA treatment, were acceptable for consumption.

Effects of food composition on the inactivation of foodborne microorganisms by chlorine dioxide.

Chlorine dioxide (ClO2) is a strong oxidizing agent that can be applied in solution as well as in the gaseous state. It has bactericidal, fungicidal and viricidal properties. Several food-related microorganisms, including Gram-negative and Gram-positive bacteria, yeasts, mould spores and Bacillus cereus spores were tested for their susceptibility to 0.08 mg/L gaseous ClO2 during 1 min at a relative humidity of 90%. In this screening, the resistance of the different groups of microorganisms towards gaseous ClO2 generally increased in the order Gram-negative bacteria, Gram-positive bacteria, yeasts and mould spores and Bacillus cereus spores. With this treatment, reductions of microbial numbers between 0.1 and 3.5 log cfu/cm2 could be achieved. The effects of the food components starch, fat, protein and NaCl on the antimicrobial activity of gaseous ClO2 were also evaluated. Soluble starch, corn oil, butter, whey protein isolate and NaCl were added in incremental concentrations to portions of an agar medium. Then, plates of the supplemented agars were inoculated with Leuconostoc mesenteroïdes at numbers of 4 log cfu/cm2 and subsequently treated with ClO2. Both soluble starch and NaCl did not have an effect on the antimicrobial efficiency of ClO2. However, butter, corn oil or whey protein in the agar almost eliminated the antimicrobial effect of ClO2. In corn oil-water emulsions treated with gaseous ClO2 the peroxide value increased significantly, indicating the formation of primary oxidation products. Similarly, a treatment with ClO2 increased the protein carbonyl content and induced the transformation of SH-groups to -S-S-groups in whey protein. The findings suggest that gaseous ClO2 will be a highly effective decontaminating agent for carbohydrate-rich foods, but that it would be less effective for the decontamination of high-protein and fatty foods.

A peculiar stimulatory effect of acetic and lactic acid on growth and fermentative metabolism of Zygosaccharomyces bailii.

Stimulatory or protective effects of organic acids at low concentrations, e.g. acetic and lactic acid, on microorganisms have previously been reported. Especially in case of Zygosaccharomyces bailii, a peculiar growth stimulation by these two acids has recently been noticed. In order to elucidate this interesting phenomenon, growth and fermentative metabolism of Z. bailii was investigated in media with low pH (pH 4.0), high sugar (15% (w/v)) and different acetic and lactic acid concentrations. At both experimental temperatures (7 and 30 degrees C), a growth stimulation in the presence of 2.5% (v/v) lactic acid was observed. Furthermore at 7 degrees C, the yeast exhibited another unusual behaviour as it grew much faster in media containing 1.25% (v/v) acetic acid than in the control (without any acid). Production of fermentative metabolites was also increased together with the enhanced growth at both temperatures. These possible stimulatory effects of acetic and lactic acid should be taken into consideration when the acids are used at low doses for food preservative purpose. Presence of the acids may stimulate Z. bailii growth and fermentative metabolism, particularly at refrigeration temperature, consequently resulting in an earlier spoilage.

Shelf-life extension of minimally processed carrots by gaseous chlorine dioxide.

Chlorine dioxide (ClO(2)) gas is a strong oxidizing and sanitizing agent that has a broad and high biocidal effectiveness and big penetration ability; its efficacy to prolong the shelf-life of a minimally processed (MP) vegetable, grated carrots (Daucus carota L.), was tested in this study. Carrots were sorted, their ends removed, hand peeled, cut, washed, spin dried and separated in 2 portions, one to be treated with ClO(2) gas and the other to remain untreated for comparisons. MP carrots were decontaminated in a cabinet at 91% relative humidity and 28 degrees C for up to 6 min, including 30 s of ClO(2) injection to the cabinet, then stored under equilibrium modified atmosphere (4.5% O(2), 8.9% CO(2), 86.6% N(2)) at 7 degrees C for shelf-life studies. ClO(2) concentration in the cabinet rose to 1.33 mg/l after 30 s of treatment, and then fell to nil before 6 min. The shelf-life study included: O(2) and CO(2) headspace concentrations, microbiological quality (mesophilic aerobic bacteria, psychrotrophs, lactic acid bacteria, and yeasts), sensory quality (odour, flavour, texture, overall visual quality, and white blushing), and pH. ClO(2) did not affect respiration rate of MP carrots significantly (alpha< or =0.05), and lowered the pH significantly (alpha< or =0.05). The applied packaging configuration kept O(2) headspace concentrations in treated samples in equilibrium and prevented CO(2) accumulation. After ClO(2) treatment, the decontamination levels (log CFU/g) achieved were 1.88, 1.71, 2.60, and 0.66 for mesophilic aerobic bacteria, psychrotrophs, and yeasts respectively. The initial sensory quality of MP carrots was not impaired significantly (alpha< or =0.05). A lag phase of at least 2 days was observed for mesophilic aerobic bacteria, psychrotrophs, and lactic acid bacteria in treated samples, while mesophilic aerobic bacteria and psychrotrophs increased parallelly. Odour was the only important attribute in sensory deterioration, but it reached an unacceptable score when samples were already rejected from the microbiological point of view. The shelf-life extension was limited to one day due to the restricted effect of the ClO(2) treatment on yeast counts. Nevertheless, ClO(2) seems to be a promising alternative to prolong the shelf-life of grated carrots.

Metabolite production of yeasts on a strawberry-agar during storage at 7 degrees C in air and low oxygen atmosphere.

Changes of different quality factors of strawberries have been described until now from a physiological point of view. Possible effects on quality caused by yeast proliferation have not been described. To elucidate the metabolic activity of yeasts (i.e. Debaryomyces melissophilus, Rhodotorula glutinis, Cryptococcus laurentii), isolated from strawberries, they were inoculated on a simulation medium of strawberries (strawberry-agar). Their activity was measured by analysing and correlating microbiological counts, metabolite concentration in the headspace as well as in the medium, and sugar consumption. The isolated yeasts from strawberries could grow on the strawberry-agar, both under air conditions and modified atmosphere (MA) conditions. The maximum count for the yeasts reached 7.5-8.5 log cfu cm(-2) (air conditions) and 5.6-6.4 log cfu cm(-2) (MA conditions). Production or consumption of a number of compounds could be detected when microbial counts reached levels between 4.7 log cfu cm(-2) and 8.5 log cfu cm(-2) depending on species and atmospheric conditions. A range of volatile organic compounds, produced by the yeasts, was detected: acetone, ethyl acetate, ethanol, isopropyl acetate, ethyl butyrate, 1-propanol, 2-methyl-1-propanol, 1-butanol, 2-methyl-1-butanol, 3-methyl-1-butanol, 1-hexanol and hexyl acetate. These compounds are able to influence the sensory properties of strawberries. A simultaneous decrease in sugar concentrations (sucrose, glucose, fructose) was observed. When ethanol reached high concentrations, ethyl acetate and ethyl butyrate were produced. This production can be attributed to a detoxification of ethanol by yeasts. The fermentative metabolism of yeasts during aerobic conditions could be explained by the Crabtree effect. As the detected volatile organic compounds produced by yeasts are also found in fresh strawberries, it can be concluded that these compounds are produced both by microbiological and physiological processes.

Volatile metabolite production of spoilage micro-organisms on a mixed-lettuce agar during storage at 7 degrees C in air and low oxygen atmosphere.

This paper describes the volatile metabolite production of spoilage bacteria (Pantoea agglomerans and Rahnella aquatilis) and spoilage yeasts (Pichia fermentans and Cryptococcus laurentii), previously isolated from mixed lettuce, on a simulation medium of shredded mixed lettuce (mixed-lettuce agar) both under air conditions and modified atmosphere (MA)-conditions at 7 degrees C. These latter conditions simulated the equilibrium modified atmosphere packaging, which is used to extend the shelf-life of shredded mixed lettuce. Besides volatile metabolites, organic acid metabolites and consumption of sugars were measured. Microbiological growth on the mixed-lettuce agar resulted in metabolite production and consumption of sugars. Bacteria and yeasts produced a range of volatile organic compounds both under air conditions and MA-conditions: ethanol, ethyl acetate, 2-methyl-1-propanol, 2-methyl-1-butanol, 3-methyl-1-butanol, 2,3-butanedione, 3-methyl-1-pentanol, 1-butanol and 1-hexanol. Under MA-conditions, 2-methyl-1-butanol, 3-methyl-1-butanol and ethanol were the first compounds that were detected in the headspace as being produced by the inoculated micro-organisms. In the case of the yeast P. fermentans, production of these compounds was detected from a count of 5.0+/-0.1 log cfu/cm(2) with a fast increase when exceeding 6.0-6.5 log cfu/cm(2). Unlike P. fermentans, the yeast C. laurentii showed a slow metabolism under MA-conditions, compared to air conditions. In the case of the bacteria, production of 2-methyl-1-butanol and 3-methyl-1-butanol was detected starting from a count of 6.7+/-0.1 log cfu/cm(2) in the case of R. aquatilis and from a count of 7.1+/-0.4 log cfu/cm(2) in the case of P. agglomerans with a fast increase when exceeding 8 log cfu/cm(2). No production of ethanol by the bacteria under MA-conditions was detected in contradiction to air conditions. It could be concluded that, if these counts are reached on the cut surfaces of shredded mixed lettuce which are simulated by the mixed-lettuce agar, sensorial quality of shredded mixed lettuce could be influenced by the microbiological production of metabolites.

Role of yeast proliferation in the quality degradation of strawberries during refrigerated storage.

Quality changes of strawberries during storage can be caused both by microbiological and physiological processes. There is little known about the possible contribution of microbiological processes to the quality degradation of strawberries. In this study, quality of strawberries during storage was evaluated by analytical and sensorial analyses. It was the aim to investigate the influence of microbiological activity on the changes of different quality factors of strawberries during storage. During storage at 7 degrees C, quality was mainly determined by the odor and by visual defects. Regarding the odor, highly microbiologically contaminated late-season strawberries packaged in air at 7 degrees C became sensorially unacceptable due to the presence of high amounts of ethyl acetate. This could be attributed to the yeast proliferation: at yeast concentrations above 5.0 log cfu/g, an increase in ethanol was detected in the headspace of the strawberries. It was shown that ethanol was converted to ethyl acetate by strawberries resulting in an unacceptable odor. In an experiment with low microbiologically contaminated early-season strawberries, not reaching the above mentioned yeast counts, less ethyl acetate was detected which resulted in strawberries that were sensorially acceptable during the whole storage period (12 days). Strawberries packaged in modified atmosphere conditions showed a different quality pattern due to the effect of decreased O2-concentrations on both microbiological and physiological processes. This paper demonstrates that also microbiological processes on strawberries should be considered as they could play an important role in the sensorial quality when interacting with physiological processes.

Relation between microbiological quality, metabolite production and sensory quality of equilibrium modified atmosphere packaged fresh-cut produce.

The quality of four types of fresh-cut produce, packaged in consumer-sized packages under an equilibrium modified atmosphere and stored at 7 degrees C, was assessed by establishing the relation between the microbial outgrowth and the corresponding production of nonvolatile compounds and related sensory disorders. In vitro experiments, performed on a lettuce-juice-agar, demonstrated the production of nonvolatile compounds by spoilage causing lactic acid bacteria and Enterobacteriaceae. Pseudomonas fluorescens and yeasts, however, were not able to produce detectable amounts of nonvolatile metabolites. The type of spoilage and quality deterioration in vivo depended on the type of vegetable. Mixed lettuce and chicory endives, leafy tissues, containing naturally low concentrations of sugars, showed a spoilage dominated by Gram-negative microorganisms, which are not producing nonvolatile compounds. Sensory problems were associated with visual properties and the metabolic activity of the plant tissue. Mixed bell peppers and grated celeriac, on the other hand, demonstrated a fast and intense growth of spoilage microorganisms, dominated by lactic acid bacteria and yeasts. This proliferation resulted in detectable levels of organic acids and the rejection by the trained sensory panel was based on the negative perception of the organoleptical properties (off-flavour, odour and taste). The applied microbiological criteria corresponded well with detectable changes in sensory properties and measurable concentrations of nonvolatile compounds, surely in the cases where lactic acid bacteria and yeasts were provoking spoilage. Consequently, the freshness of minimally processed vegetables, sensitive for outgrowth of lactic acid bacteria and yeasts (e.g., carrots, celeriac, bell peppers, mixtures with non-leafy vegetables) can be evaluated via analysis of the produced nonvolatile compounds.