Evaluation of the microbiological safety and sensory quality of rainbow trout (Oncorhynchus mykiss) processed by the sous vide method

This work evaluates the microbiological quality and sensorial characteristics of rainbow trout processed by the sous vide method under different time/temperature conditions and stored at 2°C and 10°C.The equivalent heat treatments applied in core were: 3.3 min at 90°C (PV₇₀ in core 331.64), 1.04 min at 90°C (PV₇₀ in core 104.5) and 5.18 min at 70°C (PV₇₀ in core 5.18). The product stored at 2°C or 10°C was periodically evaluated to investigate its sensory quality, microbiological condition and proximate composition. The batches stored at 2°C had lower growth counts of mesophiles and psychrotrophs. Moreover, these counts decreased by increasing the heating temperature and time. Staphylococcus aureus, Bacillus cereus, Clostridium perfringens and Listeria monocytogenes were not found in any of the samples. Neither aerobic nor anaerobic spores were detected in the trout samples processed at 90°C for 3.3 min in core and stored at 2°C after 45 days. A heat treatment of 90°C for 3.3 min in core was the most effective to extend the shelf-life of trout (>45 days).Therefore, it can be concluded that the treatment at 90°C for 3.3 min in core is the most effective to ensure the safety and extend the shelf-life of sous vide trout preserving its sensory characteristics.This study establishes the microbiological quality of sous vide trout and emphasizes the relevance of the microbiological quality of the raw materials, heat treatment and storage temperature to ensure the safety of the product, since this fatty fish is not eviscerated and high counts can be found in the raw fish.     

Considerations for post-lethality treatments to reduce Listeria monocytogenes from fully cooked bologna using ambient and pressurized steam

During processing of ready-to-eat (RTE) deli meats, any secondary processing procedures such as peeling and cutting introduce the distinct possibility of cross-contamination between equipment, personnel, and food. To eliminate or reduce pathogens such as Listeria monocytogenes and ensure food safety, RTE deli meats can be pasteurized prior to or after packaging. In this study, ambient steam in-package pasteurization was compared with pressurized steam prepackaging pasteurization to reduce L. monocytogenes from fully cooked RTE bologna. The bologna (14 cm diameter×1.5 cm thickness) samples were surface-inoculated to contain about 8 log₁₀ of L. monocytogenes. To achieve 2 log reductions for L. monocytogenes, the bologna samples needed to be treated for about 10 s in pressurized steam at 131 °C or for about 2.5 min in ambient steam at 100 °C. The pasteurization time using pressurized steam treatment was about 75–90% shorter than using ambient steam treatment. Pressurized steam treatment may be integrated into a vacuum packaging unit to effectively eradicate L. monocytogenes from RTE meats just prior to sealing the retail packages to further reduce the treatment time, avoid post-treatment recontaminations by pathogens, and improve food safety without detrimentally affecting meat quality.     

High pressure carbon dioxide combined with high power ultrasound processing of dry cured ham spiked with Listeria monocytogenes

Traditionally, thermal treatments for the inactivation of Listeria monocytogenes in meat products involve undesirable changes of the product quality. In recent years, efforts have been carried out to develop alternative methods to inactivate L. monocytogenes without affecting the product quality attributes. In this context, the feasibility of combined high pressure carbon dioxide and high power ultrasound (HPCO₂ + HPU) treatment to inactivate L. monocytogenes inoculated on the surface of dry cured ham was investigated. Inactivation data were determined at 6, 8 and 12 MPa, as a function of temperature (22, 35, 45 °C) and treatment time (0.5 to 30 min), and compared to those obtained after thermal and HPCO₂ treatments.Color, pH and acidity changes of the samples after both thermal and HPCO₂ + HPU treatments were measured and compared, sensorial profile of the treated samples was evaluated by a sensory panel and shelf-life was determined by a storage study for 4 weeks at 4 °C.The results clearly revealed that HPU alone was not able to induce any microbial inactivation while HPCO₂ + HPU treatment always assured a certain level of inactivation, variable with the process temperature used: the inactivation efficiency was demonstrated higher at 35 °C rather than 22 °C and no enhancement was observed at 45 °C compared to 35 °C. Process conditions of 12 MPa, 35 °C, at 10 W for 5 min assured inactivation to undetectable level of L. monocytogenes spiked on the surface of the product with an initial concentration of about 10⁹ CFU/g. No differences were detected between acidity, pH, color and sensory attributes of the untreated and HPCO₂ + HPU treated dry cured ham surface, while slight differences were measured between the values obtained for the untreated and thermally treated samples. Additionally, the storage study demonstrated that a full microbial and quality shelf-life was assured for 4 weeks at 4 °C. The results obtained may open the doors to the application of such an innovative process at industrial level, in particular to treat ham-type or meat products.     

Inactivation of Escherichia coli O157:H7 and Listeria monocytogenes in milk by caprylic acid and monocaprylin

Listeria monocytogenes and Escherichia coli O157:H7 are major foodborne pathogens implicated in various outbreaks involving pasteurized or unpasteurized milk, and various dairy products. The objective of this study was to determine the antibacterial effect of caprylic acid (CA, C_8:0) and its monoglyceride, monocaprylin (MC) on L. monocytogenes and E. coli O157:H7 in whole milk. A five-strain mixture of E. coli O157:H7 or L. monocytogenes was inoculated in autoclaved milk (10⁶ CFU/ml) containing 0, 25, or 50 mM of CA or MC. At 37°C, all the treatments, excepting 25 mm CA, reduced the population of both pathogens by approximately 5.0 log CFU/ml in 6 h. At 24 h of storage at 8°C, MC at both levels and CA at 50 mM decreased L. monocytogenes and E. coli O157:H7, respectively by >5.0 log CFU/ml. At 48 h of 4°C storage, populations of L. monocytogenes and E. coli O157:H7 were decreased to below detection level (enrichment negative) by 50 mm of MC and CA, respectively. Results indicate that MC could potentially be used to inhibit L. monocytogenes and E. coli O157:H7 in milk and dairy products, but sensory studies need to be conducted before recommending their use.     

Influence of composition and processing of Frankfurter sausages on the growth dynamics of Listeria monocytogenes under vacuum

Ready-to-eat (RTE) meat products, such as Frankfurter sausages, are often linked to cases of listeriosis, which is a disease caused by the foodborne pathogen Listeria monocytogenes. Frankfurter sausages vary, from manufacturer to manufacturer, in many aspects: (i) composition, (ii) physicochemical characteristics, (iii) background flora, (iv) processing technology and (v) organoleptic characteristics. Some of these factors have been widely investigated for their effect on L. monocytogenes on food products. However, studies on the specific effect of composition and processing technology on L. monocytogenes dynamics between two sausages are lacking. In this study, the growth dynamics of L. monocytogenes on two types of Frankfurter sausages, fresh and in brine, were studied at constant storage temperatures (4, 8 and 12 °C). Additionally, the physicochemical and compositional characteristics of both types of sausages were analysed. In order to study the isolated effect of preservatives, L. monocytogenes growth dynamics, at 4 °C and 30 °C, were studied in two types of liquid systems. These were prepared with the same level of preservatives as in the two types of sausages. Results indicated no major significant differences in physicochemical characteristics for the two types of sausages; but, statistically significant variability was detected in the concentration of preservatives. In liquid systems, the maximum specific growth rate (μ_max) remained unaffected by the effect of preservatives, but the lag phase was longer in the system mimicking fresh sausages. In sausages, the ‘in brine’ type had two-fold higher μ_max at all temperatures and shorter lag phase at 4 °C. The presence/absence of sausage skin, which was found to be impermeable from L. monocytogenes cells and was present in the fresh sausage, could explain those differences. In conclusion, this study highlighted the influence of processing factors, and specifically of the sausage casing on L. monocytogenes growth dynamics. Therefore, an edible membrane, which is heat resistant and impermeable to the cells, could be a hurdle strategy to control the microbiological food safety.     

PFGE characterisation and adhesion ability of Listeria monocytogenes isolates obtained from bovine carcasses and beef processing facilities

Listeria monocytogenes is a pathogen capable of adhering to many surfaces and forming biofilms, which may explain its persistence in food processing environments. This study aimed to genetically characterise L. monocytogenes isolates obtained from bovine carcasses and beef processing facilities and to evaluate their adhesion abilities. DNA from 29 L. monocytogenes isolates was subjected to enzymatic restriction digestion (AscI and ApaI), and two clusters were identified for serotypes 4b and 1/2a, with similarities of 48% and 68%, respectively. The adhesion ability of the isolates was tested considering: inoculum concentration, culture media, carbohydrate source, NaCl concentration, incubation temperature, and pH. Each isolate was tested at 10⁸ CFU mL^− 1 and classified according to its adhesion ability as weak (8 isolates), moderate (17) or strong (4). The isolates showed higher adhesion capability in non-diluted culture media, media at pH 7.0, incubation at 25 °C and 37 °C, and media with NaCl at 5% and 7%. No relevant differences were observed for adhesion ability with respect to the carbohydrate source. The results indicated a wide diversity of PFGE profiles of persistent L. monocytogenes isolates, without relation to their adhesion characteristics. Also, it was observed that stressing conditions did not enhance the adhesion profile of the isolates.     

Application of statistical process control,sampling, and validation for producing Listeria monocytogenes-free chicken leg quarters processed in steam followed by impingement cooking

Chicken leg quarters (180–230 g) were processed for 4 min in steam at 99°C and then in an air impingement oven for 24 min at an oven temperature of 232°C, an air velocity of 2 m/s, and a humidity of 60%. The cooked chicken leg quarters were sampled to measure for the end-point internal temperatures. Sampling size in each subgroup for the internal temperature measurements was determined based on a normal distribution at a confidence level of 95%. The process mean, range, and standard deviation at 95% confidence level were 73.9°C, 1.8°C, and 0.9°C, respectively, for the internal temperatures of the cooked chicken leg quarters. The process lethality was validated for up to 7  log₁₀ cfu/g reductions of Listeria monocytogenes in the cooked chicken leg quarters and verified by an inoculation study in which the chicken leg quarters were injected with 0.1 ml of the culture per cm² of the product surface area to contain 7–8 log₁₀ cfu/g of L. monocytogenes. This paper provided an approach for process control, sampling, and validation to reduce pathogens in fully cooked poultry products.     

Survival of Listeria monocytogenes in Galotyri,a traditional Greek soft acid-curd cheese,stored aerobically at 4°C and 12°C

Galotyri is a traditional Greek soft acid-curd cheese, which is made from ewes’ or goats’ milk and is consumed fresh. Because cheese processing may allow Listeria monocytogenes post-process contamination, this study evaluated survival of the pathogen in fresh cheese during storage. Portions (0.5 kg) of two commercial types (<2% salt) of Galotyri, one artisan (pH 4.0±0.1) and the other industrial (pH 3.8±0.1), were inoculated with ca. 3 or 7 log cfu g⁻¹ of a five-strain cocktail of L. monocytogenes and stored aerobically at 4°C and 12°C. After 3 days, average declines of pathogen's populations (PALCAM agar) were 1.3–1.6 and 3.7–4.6 log cfu g⁻¹ in cheese samples for the low and high inocula, respectively. These declines were independent (P>0.05) of the cheese type or the storage temperature. From day 3, however, declines shifted to small or minimal to result in 1.4–1.8 log cfu g⁻¹ of survivors at 28 days of storage of all cheeses at 4°C, indicating a strong “tailing” independent of initial level of contamination. Low (1.2–1.7 log cfu g⁻¹) survival of L. monocytogenes also occurred in cheeses at 12°C for 14 days, which were prone to surface yeast spoilage. When ca. 3 log cfu g⁻¹ of L. monocytogenes were inoculated in laboratory scale prepared Galotyri of pH ≅4.4 and ≅3% salt, the pathogen died off at 14 and 21 days at 12°C and 4°C, respectively, in artisan type cheeses fermented with the natural starter. In contrast, the pathogen survived for 28 days in cheeses fermented with the industrial starter. These results indicate that L. monocytogenes cannot grow but may survive during retail storage of Galotyri despite its low pH of or slightly below 4.0. Although contamination of Galotyri with L. monocytogenes may be expected low (<100 cfu g⁻¹) in practice, that long-term survival of the pathogen in commercial cheeses was shown to be unaffected by the artificial contamination level (3 or 7 logs) and the storage temperature (4°C or 12°C), which should be a concern.     

Growth of Listeria Monocytogenes in Traditional Austrian Meat Jelly Products

Individual components of a traditional meat jelly (cooked meat chunks, gelatin and preboiled vegetable) with differences in pH and a_w can constitute a niche for the multiplication of Listeria. Listeria monocytogenes counts remained stable in jelly over 21 days at 2 and 8 °C, whereas in meat and vegetables, a >1 log₁₀ unit increase was observed after 7 days at 2 °C (or >5 log₁₀ at 8 °C). In the composed product, Listeria numbers remained stable at 2 °C (21 days), but increased more than 1 log₁₀ during 7 days at 8 °C. Improving safety of jellied meat by lowering pH is discussed.     

The influence of lactoperoxidase,heat and low pH on survival of acid-adapted and non-adapted Escherichia coli O157:H7 in goat milk

In hot climates where quality of milk is difficult to control, a lactoperoxidase (LP) system can be applied in combination with conventional preservation treatments at sub-lethal levels to inhibit pathogenic microbes. This study investigated the effect of combined heat treatments (55 °C, 60 °C and 72 °C) and milk acidification (pH 5.0) on survival of acid-adapted and non-adapted Escherichia coli O157:H7 strains UP10 and 1062 in activated LP goat milk. Heat treatment at 72 °C eliminated E. coli O157:H7. Acid-adapted strains UP10 and 1062 cells showed resistance to combined LP and heat at 60 °C in fresh milk. The inhibition of acid-adapted and non-adapted E. coli O157:H7 in milk following combined LP-activation, heat (60 °C) and milk acidification (pH 5.0) suggests that these treatments can be applied to reduce E. coli O157:H7 cells in milk when they occur at low numbers (<5 log₁₀ cfu mL^?1) but does not eliminate E. coli O157:H7 to produce a safe product.     

Effect of mild heat pre-treatment with alkaline electrolyzed water on the efficacy of acidic electrolyzed water against Escherichia coli O157:H7 and Salmonella on Lettuce

Cut lettuce dip-inoculated with Escherichia coli O157:H7 and Salmonella was treated with alkaline electrolyzed water (AlEW) at 20°C for 5 min, and subsequently washed with acidic electrolyzed water (AcEW) at 20°C for 5 min. Pre-treatment with AlEW resulted in an approximate 1.8 log₁₀ cfu/g reduction of microbial populations, which was significantly (p⩽0.05) greater than microbial reductions resulting from other pre-treatment solutions, including distilled water and AcEW. Repeated AcEW treatment did not show a significant bacterial reduction. Mildly heated (50°C) sanitizers were compared with normal (20°C) or chilled (4°C) sanitizers for their bactericidal effect. Mildly heated AcEW and chlorinated water (200 ppm free available chlorine) with a treatment period of 1 or 5 min produced equal reductions of pathogenic bacteria of 3 log₁₀ and 4 log₁₀ cfu/g, respectively. The procedure of treating with mildly heated AlEW for 5 min, and subsequent washing with chilled (4°C) AcEW for period of 1 or 5 min resulted in 3–4 log₁₀ cfu/g reductions of both the pathogenic bacterial counts on lettuce. Extending the mild heat pre-treatment time increased the bactericidal effect more than that observed from the subsequent washing time with chilled AcEW. The appearance of the mildly heated lettuce was not deteriorated after the treatment. In this study, we have illustrated the efficacious application of AlEW as a pre-wash agent, and the effective combined use of AlEW and AcEW.     

Resistance of Campylobacter jejuni to heat and to pulsed electric fields

The resistance of Campylobacter jejuni NCTC 11351 to heat and to pulsed electric fields (PEF) was studied at different treatment intensities (temperatures between 52 and 60 °C, and electric field strengths between 15 and 35 kV/cm, respectively). The influence of the growth phase, the pH of the treatment medium and the presence of sodium pyruvate in the recovery medium was also examined. A model based on the Weibull distribution was used to describe the inactivation curves, and times for the first decimal reduction were calculated (δ values). C. jejuni cells did not increase their resistance to heat nor to PEF upon entrance into stationary phase. The acidification of the treatment medium from 7.0 to 4.5 caused a sensitization of C. jejuni to heat (δ value at 55 °C × 1/4); on the contrary, resistance to PEF was increased (δ value at 25 kV/cm × 2.5). The absence of pyruvate in the recovery medium prevented recovery of a high percentage of heat-treated cells, but did not affect PEF survival. Whereas C. jejuni can be considered a heat sensitive organism (δ value at 55 °C and buffer of pH 7.0 of 2 min, z value 4.40 °C), it showed a relatively high resistance to PEF as compared to other vegetative cells (δ value at 25 kV/cm and buffer of pH 7.0 of 7 pulses, z_PEF value 8.20 kV/cm). Results obtained in this investigation indicate that Campylobacter spp. should be taken into account for the design of PEF treatments for food hygienization.Industrial relevanceBefore PEF can be commercially implemented it is necessary to determine its efficacy on pathogenic microorganisms of interest in order to ensure safety of food. There is no data available about the resistance of C. jejuni to pulsed electric fields, although it is now recognised as the leading cause of bacterial food-borne gastroenteritis throughout the world. In this research we characterize the resistance to heat and to PEF of C. jejuni NCTC 11351. Physiological factors affecting its survival to both agents are also explored.     

Quantitative evaluation and prediction for preservation quality of cold shocked cucumber based on entropy

To evaluate and predict preservation quality of cold shocked cucumber and study the effects of cold shock parameters on the preservation quality, several entropy-based models were proposed. Cucumbers were cold shocked at different temperatures (0, 3, 6 °C) for different durations (20, 40, 60 min), and their preservation quality was evaluated by the proposed models. Results show that, the evaluation model can objectively evaluate preservation quality of cold shocked cucumber, and cold shocked at 3 °C for 40 min gets optimal preservation effect. Cold shock treatments at 0 °C and 3 °C improve preservation quality of cucumber effectively, while that at 6 °C fails. Therefore, entropy change caused by cold shock treatment should be higher than a certain critical value to improve preservation quality of cucumber effectively, and the duration is crucial for preservation quality when meeting this condition. The composite entropy change (S*) shows effect of both temperature and duration and characterizes cold shock intensity, and proposed prediction model of cold shock preservation quality can predict preservation quality of cucumber cold shocked at different temperatures and durations well.Industrial relevanceFresh fruits and vegetables contain rich essential substances to our bodies and play an active role in improving people's health. Since it has obvious seasonal regional characteristics on the fruits and vegetables production, pretreatment is very important for prolonging their storage periods. Cold shock treatment for many fruits and vegetables indicate that this method can effectively improve their preservation quality and have broad application space.This study lies in providing a series of methods based on entropy to characterize the cold shock intensity to fruits and vegetables, carrying out objective quantitative evaluation and prediction for preservation quality of the cold shock treatment, and based on which, the influence mechanism of cold shock intensity (temperature and duration) on preservation quality of fruits and vegetables was analyzed. The results will contribute to better design and optimization of the cold shock process for postharvest fruits storage.     

The inactivation of Listeria monocytogenes by pulsed electric field (PEF) treatment in a static chamber

An experimental analysis of the effect of pulsed electric field (PEF) energy on the inactivation of Listeria monocytogenes was conducted using a custom-designed static chamber and a gel suspension medium for treatment. This allowed PEF energy to be delivered to the suspension under near isothermal conditions. The effects of variations in the number of pulses (5–50 pulses), electric field strength (15–30 kV/cm), temperature (0–60°C) and media bases (water and skim milk) on the inactivation of L. monocytogenes were examined. At temperatures less than 50°C a maximum of 1 log reduction was obtained for L. monocytogenes regardless of pulse number or electric field strength within the ranges examined. In skim milk no reduction occurred. At 50°C and 55°C synergy between PEF and thermal energy was observed. The experimental approach separated the contribution of PEF and thermal energy to total kill and thus allowed this synergy to be quantified. At 55°C the kill due to PEF energy increased to 4.5 logs with another 4.5 logs reduction attributable to thermal energy. It appears that under the conditions of this study PEF alone has a very limited effect on the reduction of L. monocytogenes. However, the addition of thermal energy not only contributed to the kill, but also increased the susceptibility of L. monocytogenes to PEF energy.     

Conventional freezing plus high pressure–low temperature treatment: Physical properties,microbial quality and storage stability of beef meat

Meat high-hydrostatic pressure treatment causes severe decolouration, preventing its commercialisation due to consumer rejection. Novel procedures involving product freezing plus low-temperature pressure processing are here investigated. Room temperature (20 °C) pressurisation (650 MPa/10 min) and air blast freezing (−30 °C) are compared to air blast freezing plus high pressure at subzero temperature (−35 °C) in terms of drip loss, expressible moisture, shear force, colour, microbial quality and storage stability of fresh and salt-added beef samples (Longissimus dorsi muscle). The latter treatment induced solid water transitions among ice phases. Fresh beef high pressure treatment (650 MPa/20 °C/10 min) increased significantly expressible moisture while it decreased in pressurised (650 MPa/−35 °C/10 min) frozen beef. Salt addition reduced high pressure-induced water loss. Treatments studied did not change fresh or salt-added samples shear force. Frozen beef pressurised at low temperature showed L, a and b values after thawing close to fresh samples. However, these samples in frozen state, presented chromatic parameters similar to unfrozen beef pressurised at room temperature. Apparently, freezing protects meat against pressure colour deterioration, fresh colour being recovered after thawing. High pressure processing (20 °C or −35 °C) was very effective reducing aerobic total (2-log₁₀ cycles) and lactic acid bacteria counts (2.4-log₁₀ cycles), in fresh and salt-added samples. Frozen + pressurised beef stored at −18 °C during 45 days recovered its original colour after thawing, similarly to just-treated samples while their counts remain below detection limits during storage.     

Efficacy of plant extracts in inhibitingAeromonas hydrophilaandListeria monocytogenesin refrigerated,cooked poultry

Alcohol extracts of angelica root, banana purée, bay, caraway seed, carrot root, clove (eugenol), marjoram, pimento leaf and thyme were applied to cooked chicken to determine their antimicrobial activities against Aeromonas hydrophilaand Listeria monocytogenes.Skinless chicken breast meat was cooked to an internal temperature of 85°C, allowed to cool to c. 5°C, then treated by surface application with plant extracts. Low (10 cfu g⁻¹)or high (10⁵ cfu g⁻¹)populations of A. hydrophilaand L. monocytogeneswere applied and samples were stored at either 5 or 15°C for up to 14 or 7 days, respectively. Eugenol and pimento extracts were most effective in inhibiting growth of both bacteria. A. hydrophilawas the more sensitive to the two treatments, with 4 log₁₀ cfu g⁻¹less growth occurring at 14 days at 5°C on eugenol-treated breast meat than on control samples. These results suggested that plant extracts might be useful as antimicrobials in cooked, ready-to-eat chicken meat.     

High-pressure homogenisation of raw bovine milk. Effects on fat globule size distribution and microbial inactivation

Whole raw milk was processed using a 15 L h⁻¹ homogeniser with a high-pressure (HP) valve immediately followed by a cooling heat exchanger. The influence of homogenisation pressure (100–300 MPa) and milk inlet temperature T_in (4°C, 14°C or 24°C) on milk temperature T₂ at the HP valve outlet, on fat globule size distribution and on the reduction of the endogenous flora were investigated. The T_in values of 4–24°C led to milk temperatures of 14–33°C before the HP valve, mainly because of compression heating. High T_in and/or homogenisation pressure decreased the fat globule size. At 200 MPa, the d_4.3 diameter of fat globules decreased from 3.8±0.2 (control milk) to 0.80±0.08 μm, 0.65±0.10 or 0.37±0.07 μm at T_in=4, 14°C or 24°C, respectively. A second homogenisation pass at 200 MPa (T_in=4°C, 14°C or 24°C) further decreased d_4.3 diameters to about 0.2 μm and narrowed the size distribution. At all T_in tested, an homogenisation pressure of 300 MPa induced clusters of fat globules, easily dissociated with SDS, and probably formed by sharing protein constituents adsorbed at the fat globule surface. The total endogenous flora of raw milk was reduced by more than 1 log cycle, provided homogenisation pressure was ⩾200 MPa at T_in=24°C (T₂∼60°C), 250 MPa at T_in=14°C (T₂∼62°C), or 300 MPa at T_in=4°C (T₂∼65°C). At all T_in tested, a second pass through the HP valve (200 MPa) doubled the inactivation ratio of the total flora. Microbial patterns of raw milk were also affected; Gram-negative bacteria were less resistant than Gram-positive bacteria.     

Application of electro-activated potassium acetate and potassium citrate solutions combined with moderate heat treatment on the inactivation of Clostridium sporogenes PA 3679 spores

Combined effects of moderate temperatures and the electro-activated aqueous solutions of potassium acetate and potassium citrate on the inactivation of C. sporogenes PA 3679 spores (D_121°C = 1.18 min) were studied. Four types of solutions (potassium acetate with/without KCl and potassium citrate with/without KCl) were activated at 400 mA for 60 min. The oxidation reduction potential (ORP) and pH values ranged from + 417.50 to + 1043.33 mV and 3.18 to 3.47, respectively. The combination of these solutions with a moderate heat treatment (95 °C, 105 °C, and 115 °C) for different time (5, 10, 20, and 30 min) was sufficient to reach a 100% of spore destruction (inactivation) in a medium with an initial contamination level comprised between 7.0 and 7.8 log CFU/mL. The sporicidal effect of solutions was also present even if activated solutions were applied alone against spores without being combined with heat treatment. Spore morphology was determined under transmission electron microscopy and showed that there were important damages, such as rupture of spores and release of spore components in all of the treated spores. Thus, the sporicidal effect detected was the result of inactivation mechanisms of electro-activated solutions on spores. In almost all of observed micrographs, there were coreless spores, deformed spores, or debris of spores. The current investigation can be used for achieving further studies in order to better understand the mechanisms of inactivation of C. sporogenes spores by electro-activated solutions.Industrial relevanceThis research article aims to study the combined effect of electro-activated potassium acetate and citrate solutions and moderate heat treatment on the viability of Clostridium sporogenes in model solutions as a non-pathogenic surrogate of Clostridium botulinum. The objective was to use hurdle technology to produce nutritious, minimally processed foods while ensuring food safety. Moreover, this approach allowed for a reduced level of sodium in canned foods since the solutions were sodium-free.     

Preliminary characterization of bacteriocins from Lactococcus lactis,Enterococcus faecium and Enterococcus mundtii strains isolated from turbot (Psetta maxima)

The aim of this work was the characterization of new strains of lactic acid bacteria (LAB) from farmed fish and with potential application as biopreservatives against both Listeria monocytogenes and Staphylococcus aureus. Twenty-five strains of LAB isolated from the muscle of farmed turbot were investigated. Genetic identification of the bacteriocin-producing LAB strains was performed by means of a PCR method using novel BAL1/BAL2 16S ribosomal-RNA-targeted primers. Maximum bacteriocin production by Lactococcus lactis ssp. lactis USC-39, Enterococcus faecium USC-46 and Enterococcus mundtii USC-51 was detected in the stationary phase of growth. Both acidification and the production of hydrogen peroxide by LAB were ruled out as the source of the inhibition. In contrast, the antimicrobial activity of all three LAB strains was inactivated by the addition of proteinase K, thus confirming the proteinaceous nature of the inhibition. The activity against L. monocytogenes was maintained in the 3.5–5.5 or 3.5–6.5 pH range, depending on the LAB strain. Likewise, inhibition of S. aureus strains was observed in the 3.5–4.5 and in the 3.5–5.5 pH ranges, depending on the LAB strain and on the S. aureus strain tested. Bacteriocin activity was stable in all three strains after heating the cell-free extract for 60 min at 100 °C, or even for 15 min at 121 °C, in all the three LAB strains. The acidic and heat-resistant bacteriocins produced by the three LAB strains isolated from turbot, able to inhibit the growth of both L. monocytogenes and S. aureus may find application as biopreservatives in fermented and/or heated food products.