Effect of different freezing processes on the microstructure of Atlantic salmon (Salmo salar) fillets

Salmon fillets were frozen either by pressure shift freezing (PSF, 200 MPa, − 18 °C or 100 MPa, − 10 °C) or by air-blast freezing (ABF, − 30 °C, 1 m/s or 4 m/s) or direct-contact freezing, and then stored at − 20 °C for 6 months. The influence of these treatments on the microstructure of Salmon fillets was studied. The equivalent diameter of the intracellular ice crystals were 14.69 ± 4.11, 5.52 ± 2.11, and 30.65 ± 6.31 μm for the samples subjected respectively to PSF at 100, 200 MPa and ABF (− 30 °C, 4 m/s) after 2 days of storage. Smaller and more regular intracellular ice crystals were observed in fillets frozen by PSF (200 MPa) compared with PSF (100 MPa), ABF and direct-contact frozen ones. Significant differences were observed between the size of the ice crystals obtained after conventional freezing process and PSF. Large and extracellular ice crystals were observed in fillets frozen by ABF (1 m/s) and direct-contact frozen. Minimal changes in the size of ice crystals were observed during a 3 months storage.

Industrial relevance

This paper compares different freezing methods and subsequent frozen storage with respect to their effect on microstructures of salmon fillets. Pressure shift freezing at 200 MPa was superior to conventional freezing regarding small and regular ice crystal formation. Interestingly, during frozen storage for up to 3 months the high quality product obtained via pressure freezing at 200 MPa could be retained. For longer storage periods lower pressures (100 MPa) seem sufficient to achieve stable ice crystals.     

Green tea extract as a natural antioxidant to extend the shelf-life of fresh-cut lettuce

Green tea extract (GT) was evaluated as a preservative treatment for fresh-cut lettuce. Different quality markers, e.g. respiration, browning, ascorbic acid and carotenoid content were evaluated. GT concentration (0.25, 0.5 and 1 g 100 mL^− 1) and temperature (20 °C and 50 °C) were tested. Optimal GT treatment (0.25 g 100 mL^− 1 at 20 °C) was compared with chlorine (120 ppm at 20 °C). High GT concentrations (0.5 g 100 mL^− 1 and 1.0 g 100 mL^− 1) maintained better prevent ascorbic acid and carotenoid loss than 0.25 g 100 mL^− 1 GT and chlorine. GT increased browning of samples, probably due to the content of polyphenols of the treatment; the use of heat-shock reduced this negative effect. GT and heat-shock combined also showed negative effects, reducing the antioxidant content (ascorbic acid and carotenoids). No significant differences were observed between chlorine and optimal GT (0.25 g 100 mL^− 1 at 20 °C) in browning appearance and sensory properties. GT better kept the antioxidant activity of the samples than chlorine.

Industrial relevance

An alternative treatment for minimally processed Iceberg lettuce is tested, based on its antioxidant capacity. Minimally processed industry is constantly looking for new treatments to avoid the use of chlorine which is a standard at the moment.     

Effect of X-ray treatments on inoculated Escherichia coli O157: H7, Salmonella enterica, Shigella flexneri and Vibrio parahaemolyticus in ready-to-eat shrimp

This study was conducted to evaluate the inactivation effect of X-ray treatments on Escherichia coli O157: H7, Salmonella enteric (S. enterica), Shigella flexneri (S. flexneri) and Vibrio parahaemolyticus (V. parahaemolyticus) artificially inoculated in ready-to-eat (RTE) shrimp. A mixed culture of three strains of each tested pathogen was used to inoculate RTE shrimp. The shrimp samples were inoculated individually with selected pathogenic bacteria then aseptically placed in sterile plastic cups and air-dried at 22 °C for 30 min (to allow bacterial attachment) in the biosafety cabinet prior to X-ray treatments. The inoculated shrimp samples were then placed in sterilized bags and treated with 0.1, 0.2, 0.3, 0.5, 0.75, 1.0, 2.0, 3.0 and 4.0 kGy X-ray at ambient temperature (22 °C and 60% relative humidity). Surviving bacterial populations were evaluated using a non-selective medium (TSA) with the appropriate selective medium overlay for each bacterium; CT-SMAC agar for E. coli O157: H7, XLD for S. enterica and S. flexneri and TCBS for V. parahaemolyticus. More than a 6 log CFU reduction of E. coli O157: H7, S. enterica, S. flexneri and V. parahaemolyticus was achieved with 2.0, 4.0, 3.0 and 3.0 kGy X-ray, respectively. Furthermore, treatment with 0.75 kGy X-ray significantly reduced the initial microflora on RTE shrimp samples from 3.8 ± 0.2 log CFU g⁻¹ to less than detectable limit (<1.0 log CFU g⁻¹).     

Effect of heat and thermosonication treatments on watercress (Nasturtium officinale) vitamin C degradation kinetics

The use of ultrasound in food processing creates novel and interesting methodologies, which are often complementary to classical techniques. In this work, the effect of heat and the combined treatment heat/ultrasound (thermosonication) on the thermal degradation kinetics of vitamin C in watercress (Nasturtium officinale) was studied in the temperature range of 82.5 to 92.5 °C. First order reaction kinetics adequately described the vitamin C losses during both blanching processes.The activation energies and the reaction rates at 87.5 °C for heat (H) and thermosonication (Ts) treatments were, respectively, E_avitCH = 150.47 ± 42.81 kJ mol^− 1 and E_avitCTs = 136.20 ± 60.97 kJ mol^− 1, and k_{87.5 °CvitCH} = 0.75 ± 0.10 min^− 1 and k_{87.5 °CvitCTs} = 0.58 ± 0.11 min^− 1. No significant differences (P > 0.05) were detected between both treatments. The thermosonication treatment was found to be a better blanching process, since it inactivates watercress peroxidase at less severe blanching conditions and consequently retains vitamin C content at higher levels. The present findings will help to optimise the blanching conditions for the production of a new and healthy frozen product, watercress, with heat and a new blanching process methodology.

Industrial relevance

Thermosonication blanching can be useful since it reduces processing times, and consequently minimizes the adverse effects of heating on watercress quality. This new application will provide good material, in terms of vitamin C, for further processes, and can be an excellent alternative to the traditional heat treatment.     

Effect of modified atmosphere and temperature abuse on the growth from spores and cereulide production of Bacillus weihenstephanensis in a cooked chilled meat sausage

The effect of modified atmosphere packaging (MAP) on the germination and growth of toxin producing psychrotolerant Bacillus spp is not well described. A model agar system mimicking a cooked meat product was used in initial experiments. Incubation at refrigeration temperature of 8 °C for 5 weeks of 26 Bacillus weihenstephanensis including two emetic toxin (cereulide) producing strains showed that B. weihenstephanensis is sensitive to MAP containing CO₂. The sensitivity to 20% CO₂ was dependent on strain and oxygen level, being increased when oxygen was excluded from the MAP. Growth from spores was observed at the earliest within 2 weeks when 20% CO₂ was combined with 2% O₂ and in 3 weeks when combined with “0”% O₂ (the remaining atmosphere was made up from N₂). Results were validated in a cooked meat sausage model for two non-emetic and one emetic B. weihenstephanensis strain. The packaging film oxygen transfer rates (OTR) were 1.3 and 40 ml/m²/24 h and the atmospheres were 2% O₂/20% CO₂ and “0”% O₂/20% CO₂. Oxygen availability had a large impact on the growth from spores in the MAP meat sausage, only the most oxygen restricted condition (OTR of 1.3 ml/m²/24 h and “0”% O₂/20 % CO₂) inhibited growth of the three strains during 4 weeks storage at 8 °C. Cereulide production was undetectable during storage at 8 °C irrespective of choice of the MAP (quantified by liquid chromatography mass spectrometry/mass spectrometry). MAP storage at 8 °C for 1 and 3 weeks followed by opening of packages and temperature abuse for 1.5 h daily at 20 °C during 1 week resulted in increased cell counts and variable cereulide production in the meat sausage. A pre-history at 8 °C for 1 week in MAP with OTR of 1.3 or 40 ml/m²/24 h and 2% O₂ resulted in cereulide concentrations of 0.816 – 1.353 µg/g meat sausage, while a pre-history under the most oxygen restricted condition (OTR of 1.3 ml/m²/24 h, “0”% O₂/20 % CO₂) resulted in minimal cereulide production (0.004 µg/g meat sausage) at abuse condition. Extension of MAP storage at 8 °C for 3 weeks followed by abuse resulted in a substantially reduced cereulide production.Data demonstrates that MAP can be used to inhibit growth of a psychrotolerant toxin producing Bacillus spp. during chill storage at 8 °C, and substantially reduce the risk of emetic food poisoning at abuse condition. Results are of relevance for improving safety of ready to eat processed chilled foods of extended durability.     

Ultra-high pressure homogenization treatment combined with lysozyme for controlling Lactobacillus brevis contamination in model system

It was studied the reduction of a population of Lactobacillus brevis in phosphate buffer by the combination of ultra-high pressure homogenization (UHPH) treatment and the use of lysozyme as antimicrobial. The minimum inhibitory concentration (MIC) of lysozyme against L. brevis was 50 mg.L^− 1. The observed inhibition was transitory and the growth curve showed that lysozyme addition was able to delay the L. brevis growth for 2 days. Lysozyme added at MIC concentration to a suspension of L. brevis caused a reduction of 1 logarithmic cycle after two hours of contact. The UHPH treatment against L. brevis resulted in a reduction of 7 log cycles at 200 MPa. Lysozyme was resistant to UHPH (200 MPa), without loss of muramidase activity or significant loss of antimicrobial power.A combined treatment was applied using 50 mg.L^− 1 of lysozyme and pressure between 150 and 170 MPa; the combined effect showed a reduction of about 6 logarithmic cycles and the unaltered count of L. brevis after pressure treatment for a week, with the samples stored at room temperature (25 °C).

Industrial relevance

Ultra-high pressure homogenization is an alternative process for cold pasteurization that can be used to inactivate the common spoilage microorganisms of the juice and beer industry, for example, L. brevis. Combined treatment with lysozyme increases the pressure effect and, consequently, reduces the level of pressure required for treatment. The possibility of operating with lower pressures requires less robust equipment, which makes the utilization of lower-cost equipment possible.     

Application of Kubelka–Munk analysis to the study of translucency in fresh-cut tomato

In order to assess the development of translucency in fresh-cut tomato (Lycopersicum esculentum cv. Belissimo) during refrigerated storage, two experiments were conducted. In the first one, tomato slices obtained from fruits at breaker and at red stage were stored at 5 ± 0.5 °C and monitored at regular intervals for 9 days. In the second one, slices obtained from fruits at the light-red stage were stored at 5 ± 0.5 °C, 9 ± 0.7 °C and 13 ± 0.7 °C for 4 days. Intact (control) fruits were stored at the same conditions and sliced immediately before the evaluations. In both experiments, translucency was assessed using Kubelka–Munk analysis and through visual evaluation using a scale from 0 to 4. The translucency of cut tomato slices increased during storage in both experiments. Fruits at red stage got translucent faster than fruits at breaker stage and the intensity of translucency was also higher for more ripe fruit. The storage temperature did not influence significantly the development of translucency, indicating that the water soaking of the pericarp tissue is not a result of chilling injury. The K/S (absorption coefficient/scattering coefficient) ratio increased during storage for cut fruits and remained practically constant for intact fruits, reflecting the effects of treatment observed visually. Additional experiments indicated that the removal of the locular gel combined or not with washing and drying the slice cut surface inhibited the development of translucency.

Industrial relevance

The development of translucency is a common alteration in the appearance of fresh-cut fleshy tissue after processing, which renders the product unappealing for consumption. It is of interest for the fresh-cut industry to be able to assess the development of translucency and then evaluate the effect of different procedures in its occurrence and intensity. This paper presents the use of Kubelka–Munk analysis, a technique of widespread use in other fields like printing and painting, to assess the development of translucency in sliced tomato and to study how this process is affected by storage temperature and maturity stage of the fruit.     

Heat-adaptation induced thermotolerance in Staphylococcus aureus: Influence of the alternative factor σ

The role of σ^B in the Staphylococcus aureus heat-shock induced thermotolerance was investigated. Survival curves at 58 °C of S. aureus strain Newman and its isogenic ΔsigB mutant were obtained for native and heat-shocked cells (45 °C for 5–120 min) in exponential and stationary phase of growth. The magnitude of the acquisition of thermotolerance at 58 °C depended on the growth phase and on the duration of the heat shock. Stationary growth phase cells were always more heat tolerant than exponentially growing cells and thermotolerance increased with heat-shock duration up to 120 min. S. aureus cells were able to increase their heat tolerance in the absence of the σ^B factor. In stationary phase, whereas in the parental strain the thermotolerance was increased by a factor of 12 after a heat shock of 120 min at 45 °C (δ values at 58 °C for native and heat-shocked cells were 0.63 and 7.22 min, respectively), in the mutant strain it increased 43 fold (δ values 0.09 and 3.87 min). The addition of chloramphenicol to the adaptation medium resulted in a lower increase in heat tolerance but did not prevent it completely, suggesting that S. aureus can partially increase its thermotolerance without “de novo” protein synthesis. Both the number of non-damaged cells and the proportion of cells able to repair sublethal damage were higher for heat-shocked cells.     

Comparative analysis of acid resistance in Listeria monocytogenes and Salmonella enterica strains before and after exposure to poultry decontaminants. Role of the glutamate decarboxylase (GAD) system

Data on the ability of chemical poultry decontaminants to induce an acid stress response in pathogenic bacteria are lacking. This study was undertaken in order to compare the survival rates in acid broths of Listeria monocytogenes and Salmonella enterica strains, both exposed to and not exposed to decontaminants. The contribution of the glutamate decarboxylase (GAD) acid resistance system to the survival of bacteria in acid media was also examined. Four strains (L. monocytogenes serovar 1/2, L. monocytogenes serovar 4b, S. enterica serotype Typhymurium and S. enterica serotype Enteritidis) were tested before (control) and after exposure to trisodium phosphate, acidified sodium chlorite, citric acid, chlorine dioxide and peroxyacids (strains were repeatedly passed through media containing increasing concentrations of a compound). Stationary-phase cells (10⁸ cfu/ml) were inoculated into tryptic soy broth (TSB) acidified with citric acid (pH 2.7 and 5.0) with or without glutamate (10 mM) added, and incubated at 37 °C for 15 min. Survival percentages (calculated from viable colonies) varied from 2.47 ± 0.67% to 91.93 ± 5.83%. L. monocytogenes cells previously exposed to acid decontaminants (citric acid and peroxyacids) showed, when placed in acid TSB, a higher (P < 0.05) percentage of survival (average 38.80 ± 30.52%) than control and pre-exposed to non-acidic decontaminants strains (22.82 ± 23.80%). Similar (P > 0.05) survival percentages were observed in previously exposed to different decontaminants and control Salmonella strains. The GAD acid resistance system did not apparently play any role in the survival of L. monocytogenes or S. enterica at a low pH. This study demonstrates for the first time that prior exposure to acidic poultry decontaminants increases the percentage of survival of L. monocytogenes exposed to severe acid stress. These results have important implications for the meat industry when considering which decontaminant treatment to adopt.     

Inhibition of Bacillus cereus and Bacillus weihenstephanensis in raw vegetables by application of washing solutions containing enterocin AS-48 alone and in combination with other antimicrobials

Enterocin AS-48 is a broad-spectrum cyclic antimicrobial peptide produced by Enterococcus faecalis. In the present study, the bacteriocin was tested alone and in combination with other antimicrobials for decontamination of Bacillus inoculated on alfalfa, soybean sprouts and green asparagus. Washing with enterocin AS-48 solutions reduced viable cell counts of Bacillus cereus and Bacillus weihenstephanensis by 1.0–1.5 and by 1.5–2.38 log units right after application of treatment, respectively. In both cases, the bacteriocin was effective in reducing the remaining viable population below detection levels during further storage of the samples at 6 °C, but failed to prevent regrowth in samples stored at 15 or 22 °C. Application of washing treatments containing enterocin AS-48 in combination with several other antimicrobials and sanitizers (cinnamic and hydrocinnamic acids, carvacrol, polyphosphoric acid, peracetic acid, hexadecylpyridinium chloride and sodium hypochlorite) greatly enhanced the bactericidal effects. The combinations of AS-48 and sodium hypochlorite, peracetic acid or hexadecylpyridinium chloride provided the best results. After application of the combined treatments on alfalfa sprouts contaminated with B. cereus or with B. weihenstephanensis, viable bacilli were not detected or remained at very low concentrations in the treated samples during a 1-week storage period at 15 °C. Inhibition of B. cereus by in situ produced bacteriocin was tested by cocultivation with the AS-48 producer strain E. faecalis A-48-32 inoculated on soybean sprouts. Strain A-48-32 was able to grow and produce bacteriocin on sprouts both at 15 and 22 °C. At 15 °C, growth of B. cereus was completely inhibited in the cocultures, while a much more limited effect was observed at 22 °C. The results obtained for washing treatments are very encouraging for the application of enterocin AS-48 in the decontamination of sprouts. Application of washing treatments containing AS-48 alone can serve to reduce viable cell counts of bacilli in samples stored under refrigeration, while application of combined treatments should be recommended to avoid proliferation of the surviving bacilli under temperature-abuse conditions.     

Effect of water activity and temperature on growth of Alternaria alternata on a synthetic tomato medium

Alternaria alternata is a toxigenic fungus, predominantly responsible for Blackmould of ripe tomato fruits, a disease frequently causing substantial losses of tomatoes, especially those used for canning. The objective of this study was to determine the effect of water activity (a_w, 0.904, 0.922, 0.954, 0.982) and temperature (6, 15, 21 and 35 °C) on germination and radial growth rate on a synthetic tomato medium of a cocktail inoculum of five strains of A. alternata isolated from tomato fruits affected by Blackmould. The shortest germination time (1.5 days) was observed at 0.982 a_w, both at 21 °C and 35 °C. The germination time increased with a reduction on a_w. The fastest growth rate was registered at 0.982 a_w and 21 °C (8.31 mm/day). Growth rates were higher when a_w increased. No growth or germination was observed at the lowest a_w level evaluated (0.904) after 100 days of incubation at 6 °C and 15 °C. A temperature of 6 °C caused a significant reduction in growth rates, even at the optimum a_w level. The knowledge on the ecophysiology of the fungus in this substrate is necessary to elaborate future strategies to prevent its development and evaluate the consumer health risk.     

Migration of α-tocopherol from an active multilayer film into whole milk powder

Antioxidant active packaging is a promising technology for whole milk powder (WMP) protection. In this study, the migration of α-tocopherol from a multilayer active packaging (made of high density polyethylene, ethylene vinyl alcohol and a layer of low density polyethylene containing the antioxidant) to WMP was studied. A model based on the Fick’s diffusion equation was used to calculate the diffusion coefficients (D) of α-tocopherol as 2.34 × 10⁻¹¹, 3.06 × 10⁻¹¹, and 3.14 × 10⁻¹¹ cm² s⁻¹ at 20, 30 and 40 °C, respectively. The D at 20 °C was different from those at 30 and 40 °C (P < 0.05); but it was similar at 30 and 40 °C. This low influence of temperature on the migration of α-tocopherol from 20 to 40 °C assures the release at real storage and commercialization conditions in regions with warm/hot climate. The antioxidant delivering system delayed the lipid oxidation of WMP and it was more effective at 30 and 40 °C since the rate of oxidative reactions was higher at these temperatures than at 20 °C.     

Partial nitritation treatment of underground brine waste with high ammonium and salt content

Underground brine waste containing high concentrations of ammonium and with a salinity of 3% is usually generated during the production of methane gas and iodine in the gas field of Chiba Prefecture, Japan. In this study, one swim-bed reactor, packed with a novel acrylic fiber biomass carrier (Biofringe), was applied to the partial nitritation treatment of this kind of underground brine waste. A stable nitrite production rate of 1.6 kg NO₂-N m^− 3 d^− 1 was obtained under a nitrogen loading rate of 3.0 kg-N m^− 3 d^− 1, at a pH of 7.5 and a temperature of 25 °C. Nitrate production was negligible and the effluent NO₂-N/NO_x-N ratio was above 98% due to the successful inhibition of nitrite-oxidizing bacterial activity. Free ammonia was considered to be the main factor for inhibiting the activity of nitrite-oxidizing bacteria. A microbial community shift was demonstrated by 16S rRNA analysis, and it was shown that the ammonium-oxidizing bacteria became the predominant species after successful nitrite accumulation was observed.     

Purification, characterization, and gene cloning of sphingomyelinase C from Streptomyces griseocarneus NBRC13471

Sphingomyelinase C (SMC) was purified to homogeneity from the culture supernatant of Streptomyces griseocarneus NBRC13471. The purified enzyme appeared as a single band of 38 kDa by using an electropherogram trace. The molecular mass of the enzyme as determined by MALDI-TOF MS was 32,102 Da, indicating that SMC is monomeric in nature. Under experimental conditions, the highest enzyme activity was found at pH 9.0 and 50–55 °C, and the enzyme was stable from pH 5 to 10 and up to 37 °C. The SMC activity requires Mg²⁺ or Mn²⁺ and the order of potency to enhance the activity was Zn²⁺ ≥ Mn²⁺ > Cu²⁺ ≥ Fe²⁺. Phenylmethylsulfonyl fluoride and EDTA inhibited the enzyme activity, showing that SMC belongs to a group of metalloenzymes and a class of serine hydrolases. The enzyme activity was inhibited by DTT, but not by mercaptoethanol and iodoacetamide. SDS inhibited the enzyme activity; by contrast, Triton X-100 stimulated the activity. The N-terminal and internal amino-acid sequences were determined as H₂N-APAAATPSLK, AREIAAAGFFQGND, and NTVVQETSAP. The gene encoding SMC consisted of 1020 bp encoding a signal peptide of 42 amino acids and a mature protein of 297 amino acids with a calculated molecular mass of 32,125 Da. The conserved region of DNase I-like family enzymes and the amino acid residues that are highly conserved in the active center of other bacterial SMCs were also found in the deduced amino acid sequence of S. griseocarneus SMC.     

Environmental factors modify carbon nutritional patterns and niche overlap between Aspergillus flavus and Fusarium verticillioides strains from maize

This study examined the utilization patterns of key carbon sources (CS, 24: including key sugars, amino acids and fatty acids) in maize by strains of Aspergillus flavus and Fusarium verticillioides under different water activity (a_w, 0.87–0.98 a_w) and temperature (20–35 °C) values and compared the niche overlap indices (NOI) that estimate the in vitro CS utilization profiles [Wilson, M., Lindow, S.E., 1994. Coexistence among epiphytic bacterial populations mediated through nutritional resource partitioning. Applied and Environmental Microbiology 60, 4468–4477.]. The ability to grow in these key CS in minimal media was studied for 120 h in 12 h steps. The NOI was calculated for inter-species (F. verticillioides–A. flavus) and for intra-species (A. flavus–A. flavus) using CS utilization patterns over the range of interacting environmental conditions. 30 °C, over the whole a_w range examined, was found to be optimal for utilization of the maximum number of CS by A. flavus. In contrast, for F. verticillioides this was more so at 20 °C; 25 °C allowed a suboptimal usage of CS for both species. NOIs confirmed the nutritional dominance of A. flavus at 30 °C, especially at lower a_w levels and that of F. verticillioides at 20 °C, mainly at 0.95 a_w. In other conditions of a_w, based on CS utilization patterns, the data indicated that A. flavus and F. verticillioides occupied different ecological niches. The variability in nutritional sources utilization between A. flavus strains was not related to their ability to produce aflatoxins (AFs). This type of data helps to explain the nutritional dominance of fungal species and strains under different environmental conditions. This could be useful in trying to find appropriate natural biocontrol microorganisms to compete with these mycotoxigenic species.     

Biochemical and thermal characterization of crude exo-polygalacturonase produced by Aspergillus sojae

Crude exo-polygalacturonase enzyme (produced by Aspergillus sojae), significant for industrial processes, was characterized with respect to its biochemical and thermal properties. The optimum pH and temperature for maximum crude exo-polygalacturonase activity were pH 5 and 55 °C, respectively. It retained 60–70% of its activity over a broad pH range and 80% of its initial activity at 65 °C for 1 h. The thermal stability study indicated an inactivation energy of E_d = 152 kJ mol⁻¹. The half lives at 75 and 85 °C were estimated as 3.6 and 1.02 h, respectively. Thermodynamic parameters, ΔH^*, ΔS^* and ΔG^*, were determined as a function of temperature. The kinetic constants K_m and V_max, using polygalacturonic acid as substrate, were determined as 0.424 g l⁻¹ and 80 μmol min⁻¹, respectively. SDS-PAGE profiling revealed three major bands with molecular weights of 36, 53 and 68 kDa. This enzyme can be considered as a potential candidate in various applications of waste treatment, in food, paper and textile industries.     

Effect of Zataria multiflora Boiss. essential oil and nisin on Salmonella typhimurium and Staphylococcus aureus in a food model system and on the bacterial cell membranes

The effects of different concentrations of Zataria multiflora Boiss. essential oil (EO: 0, 5, 15 and 30 μl 100 ml⁻¹) and nisin (N: 0, 0.25 and 0.5 μg ml⁻¹), temperatures (T: 25 and 8 °C), and storage times (up to 21 days) on growth of Salmonella typhimurium and Staphylococcus aureus in a commercial barley soup were evaluated in a factorial design study. The growth of S. typhimurium was significantly (P < 0.05) decreased by EO concentrations and their combinations with N concentrations at 8 °C. For S. aureus, the viable count was significantly (P < 0.05) inhibited by EO and N concentrations and their combinations, incubated at both storage temperatures. The mechanism of the antimicrobial action of EO, N, and their combinations against cell membranes of the tested organisms were also studied by measurement of the release of cell constituents and by the electronic microscopy observations of the cells. The significant increase of the cell constituents’ release of both organisms was observed as a result of treatments with EO and EO in combination with N. Electronic microscopy observations revealed that the cell membranes of S. typhimurium treated by EO and EO in combination with N were significantly damaged, while cells treated with only N looked similar to untreated cells. The electron micrographs of treated cells of S. aureus with EO, N, and their combination also showed important morphological damages and disrupted membranes.     

Preliminary screening of Bifidobacteria spp. and Pediococcus acidilactici in a Swiss cheese curd slurry model system: Impact on microbial viability and flavor characteristics

A Swiss cheese curd slurry model system was used as a preliminary screening method to determine the feasibility of the incorporation of probiotic bacteria (Bifidobacterium breve R0070, Bifidobacterium infantis R0033, Bifidobacterium longum R0175, and Pediococcus acidilactici R1001) into Swiss cheese. The cheese curd was inoculated with probiotic bacteria (8.0 log₁₀ cfu g⁻¹) and ripened anaerobically for 0, 7, and 10 days at 37 °C. Following ripening, counts of the probiotic bacteria increased to 9–10 log₁₀ cfu g⁻¹, with no significant difference in the viability of the four probiotic bacteria. Viable populations of Swiss cheese background microflora in the presence of each probiotic culture were comparable with the control. Ripening time, and to a lesser extent probiotic treatment had a significant effect on the content of several volatile flavor compounds. Similarly, ripening time contributed to a significant increase in the content of a majority of the free amino acids. The study demonstrated the feasibility of the incorporation of probiotic bacteria into Swiss cheese to produce a functional food.     

Ascospore inactivation and germination by high pressure processing is affected by ascospore age

The effects of age on high pressure resistance of the ascospores of heat resistant moulds Byssochlamys fulva, B. nivea, Neosartorya fischeri and N. spinosa were determined. Ascospores were harvested from cultures grown for 3–15 weeks at 30 °C on malt extract agar. Following filtration and determination of concentration, the ascospores were subjected to high pressure processing (HPP) at 600 MPa for 10 min in 0.1 M citrate phosphate buffer (pH 4 and 6) and mango puree (pH 5). The results supported our hypothesis that age (maturity) affects high pressure resistance of ascospores of heat resistant moulds. A reduction of log₁₀ 2.5 cfu mL^− 1 was achieved for three week old ascospores ofB. nivea whereas for nine week old ascospores only a half log reduction was achieved. Similar results were observed for B. nivea and N. fischeri. The HPP treatment caused activation of ascospores of N. spinosa, with older ascospores showing increased activation.

Industrial relevance

The observation of activation of some ascospores by HPP, indicates that HPP alone is insufficient for elimination of these problematic spoilage microorganisms. HPP would need to be combined with other hurdles in order to produce high quality pressure-treated shelf-stable fruit products.