Clean labelling sodium nitrite at pilot scale: In-situ reduction of nitrate from plant sources and its effects on the overall quality and safety of restructured cooked ham.

Growing health and environmental concerns have increased demand for all-natural products, with a focus on clean labelling. Sodium nitrite is the most widely used additive in the meat industry because it imparts the typical cured flavour and colour to meat products and, most importantly, their microbiological safety. However, due to health concerns, the European Commission is proposing revised regulations to reduce nitrate and nitrite levels in meat products. As a result, the meat industry is actively seeking alternatives. This study explored the production of four cooked hams utilising nitrate-rich vegetable sources combined with two different nitrate-reducing commercial food cultures, alongside a control ham prepared with sodium nitrite (150 ppm). Microbiological, physico-chemical (pH, water activity, nitrate and nitrite concentration, lipid profile, lipid oxidation) and sensory (texture and colour profile) characterisation of the products was carried out. Challenge tests for Listeria monocytogenes, Clostridium sporogenes and Clostridium perfringens have been performed to assess the growth of pathogens, if present in the products. Results revealed comparable microbiological and physico-chemical profiles across ham formulations, with minor differences observed in colour parameters for sample C. The sensory analysis showed that for the pilot ham formulations A and D, there were no significant differences in consumer perception compared to the control ham. In the challenge tests, L. monocytogenes levels were similar in both control and tested hams. There were no significant differences in C. sporogenes and C. perfringens counts at any temperature or between test and control samples. These results indicate that this technology has a potential future in the cured meat sector, as regulators mandate the reduction of added synthetic chemicals and consumers seek healthier and more natural ingredients in their daily diets.

Application of Ultrasound Treatments in the Processing and Production of High-Quality and Safe-to-Drink Kiwi Juice.

This study explores the potential of thermosonication as an alternative to traditional heat treatments, such as pasteurization, in the processing of fruit juices. Conventional methods often lead to undesirable quality changes in fruit juices, whereas thermosonication offers promising results regarding microbial inactivation and quality preservation. This work focused on the inactivation kinetics of Listeria innocua 2030c, a surrogate for pathogenic L. monocytogenes, in kiwifruit juice using thermosonication at 45 °C, 50 °C, and 55 °C. These treatments were compared with equivalent heat treatments. Quality attributes of the juice were also evaluated to assess process efficiency. Survival data of L. innocua were fitted with the Weibull model, estimating first decimal reduction times (δ) and shape parameters (n). The results reveal temperature and process dependencies on δ, while n remains mostly temperature and treatment independent. Thermosonication outperforms heat treatment, achieving higher L. innocua reductions while retaining quality attributes like pH, soluble solid content, and total phenolics and chlorophylls. Thermosonication at 55 °C stands out, providing a 6.2-log-cycle reduction in just 3 min with superior quality retention. These findings highlight the synergistic effect of temperature and ultrasound, making mild heat processes feasible while enhancing product quality. Thermosonication, particularly at 55 °C, emerges as an effective alternative to traditional thermal treatments for fruit juices, offering improved microbial safety without compromising product quality.

New Approaches for Improving the Quality of Processed Fruits and Vegetables and Their By-Products.

The 2030 Sustainable Development Agenda calls for all social actors to contribute to significant societal and environmental issues [...].

Thermosonication Applied to Kiwi Peel: Impact on Nutritional and Microbiological Indicators.

The peels of many fruits are rich sources of nutrients, although they are not commonly consumed. If they are properly decontaminated, they can be used as healthy food ingredients reducing food waste. The objective was to apply thermosonication processes to kiwi peel and evaluate the impact on Listeria innocua survival (a non-pathogenic surrogate of L. monocytogenes) and key nutrients and quality indicators: proteins, fibers, minerals (Ca, K, Mg, Na, and P), chlorophylls, and phenolic contents. Kiwi peels were artificially inoculated with L. innocua and thermal and thermosonication treatments were performed at 55 °C and 60 °C for 30 and 15 min maximum, respectively. Bacteria were enumerated through treatment time, and quality indicators were assessed before and at the end of treatments. A Weibull model with a decimal reduction time (D-value) was successfully used in L. innocua survival data fits. Results showed that coupling temperature to ultrasound had a synergistic effect on bacteria inactivation with significant decreases in D-values. Thermosonication at 60 °C was the most effective in terms of protein, fiber, chlorophylls, and phenolics retention. Minerals were not significantly affected by all treatments. Applying thermosonication to kiwi peel was more effective for decontamination than thermal treatments at the same temperature while allowing the retention of healthy compounds.

Freeze-Drying Processes Applied to Melon Peel: Assessment of Physicochemical Attributes and Intrinsic Microflora Survival during Storage.

Melon peel is recognized as a source of healthy nutrients and oxidant compounds. Being considered a non-edible part with no profit value, large amounts of melon rinds are discharged by fruit industries. Innovative food ingredients with potential health benefits may arise if these parts were conveniently transformed. The objective was to freeze-dry small melon peel cubes to attain a potential edible matrix. An ozone pre-treatment was applied seeking decontamination purposes and quality retention. The effect of these processes was assessed in terms of physicochemical parameters (moisture content, water activity and color), bioactive compounds (total phenolics, vitamin C and chlorophylls) and antioxidant capacity, during 7 weeks of storage at room temperature. Intrinsic microflora (mesophylls, yeasts and molds) were also monitored. Results showed that the freeze-drying process allowed retention of the most bioactive compounds analyzed, except for total phenolic content. In this case, the ozone pre-treatment was important for phenolics preservation. During the storage period, ozonated samples presented a higher content of bioactive compounds. In terms of microflora, the ozone and freeze-drying effects were not significant. Freeze-drying proved to be a suitable preservation method for melon peel. The ozone impact was not relevant in terms of decontamination.

Particle Size Effect of Integral Carob Flour on Bioaccessibility of Bioactive Compounds during Simulated Gastrointestinal Digestion.

Carob fruit is native to the Mediterranean region and produced mainly in Portugal, Italy, Morocco and Turkey. The production of the carob fruit in Portugal is highly extensive and sustainable. Currently, carob flour (CF) production is mainly achieved after pulp separation, despite it having been demonstrated that the seeds improve the extraction efficiency of bioactive compounds such as polyphenols, promoting human health. This study aimed to produce an integral CF through an innovative process and assess its physicochemical and bioactive properties at different particle sizes throughout simulated gastrointestinal tract (GIT) digestion. The sugar content profile obtained throughout GIT digestion indicated that sucrose, the sugar present at the highest concentration in undigested CF, was digested and broken down into simple sugars, namely glucose and fructose. The total phenolic content (TPC) and antioxidant activity obtained for the ≤100 µm fraction were in accordance and gastric digestion promoted an increase in the TPC value compared to the undigested sample. The >100 µm fractions displayed a distinct profile from the ≤100 µm fraction. This study showed that the particle size affects the sugar, antioxidant and total phenolic content of CFs and also their gastrointestinal tract digestion. The ≤100 µm fraction demonstrated the most suitable profile as a functional food ingredient.

Effect of Gaseous Ozone Process on Cantaloupe Melon Peel: Assessment of Quality and Antilisterial Indicators.

Fruit waste parts, particularly peel, are abundant sources of bioactive compounds. To be included in the formulation of value-added foods, peel needs to be transformed and subjected to a preservation process. Therefore, this study seeks to assess the effect of ozone on the quality and antilisterial indicators of cantaloupe melon peel paste, aiming at obtaining a product with the potential to be used as a food additive. Ozone was bubbled during 30 and 60 min, and some physicochemical characteristics (soluble solids content, pH and colour), bioactive compounds (total phenolics, chlorophylls and vitamin C) and antioxidant activity were analysed. Peel was also inoculated with Listeria innocua, used as a treatment efficiency indicator. The results indicated that, although ozone negatively affected antioxidant activity, it positively influenced all bioactive compounds analysed. An L. innocua reduction of 1.2 log cycle was achieved after ozone exposure. Ozone should be exploited as a promising technology to assure the quality/safety of cantaloupe melon peel. Indeed, if melon peel is conveniently converted into a suitable form that can be used as a food ingredient, this will promote the valorisation of waste materials with the consequent reduction of industrial by-products and new perspectives for market opportunities.

Food Consumption Determinants and Barriers for Healthy Eating at the Workplace-A University Setting.

BACKGROUND: A wide variety of social, cultural and economic factors may influence dietary patterns. This work aims to identify the main determinants of food consumption and barriers for healthy eating at the workplace, in a university setting. METHODS: A cross-sectional observational study was conducted with 533 participants. Data were obtained through the application of a self-administered questionnaire that included socio-demographic information, food consumption determinants and the main perceived barriers for healthy eating at the workplace. RESULTS: The respondents identified "price" (22.5%), "meal quality" (20.7%), and "location/distance" (16.5%). For women, the determinant "availability of healthy food options" was more important than for men (p < 0.001). The food consumption determinants at the workplace most referred to by respondents were related to the nutritional value. Smell, taste, appearance and texture, and good value for money, were also considered important for choosing food at the workplace. Respondents referred to work commitments and lack of time as the main barriers for healthy eating at the workplace. CONCLUSIONS: Identification of determinants involved in food consumption, and the barriers for healthy eating, may contribute to a better definition of health promotion initiatives at the workplace aiming to improve nutritional intake.

Impact of exposure to cold and cold-osmotic stresses on virulence-associated characteristics of Listeria monocytogenes strains.

The objective of this work was to investigate the effect of stress conditions frequently encountered in food-associated environments on virulence-associated characteristics of eight strains of Listeria monocytogenes. Strains were grown at low (11 °C, cold stress) and optimal (37 °C) temperatures and in high NaCl concentrations (6% NaCl, 11 °C; cold-osmotic stress) and tested for their ability to invade the human intestinal epithelial Caco-2 cells. Results demonstrate that the correlation between exposure to cold stress and increased invasion phenotype is strain-dependent as strains investigated exhibited different behaviours, i.e. exposure to cold stress conditions resulted in a significant increase of invasion levels in five out of the eight strains tested, when compared to growth under optimal conditions. On the other hand, when these cold-adapted cells were subsequently submitted to high salt concentrations and low temperature, their enhanced ability to invade Caco-2 was lost. Surprisingly, saturated fatty acids (SFA) and branched chain fatty acids (BCFA) decreased when L. monocytogenes were exposed to stress conditions as opposed to what has been observed in other studies, therefore highlighting that further studies will need to deepen in the understanding of the lipid metabolism of these strains. The effect of stress conditions on the survival of three selected L. monocytogenes strains through an in vitro gastrointestinal (GI) tract digestion model was further investigated. The exposure to cold-osmotic stress increased the survival of one strain through the GI tract.

Biopreservation approaches to reduce Listeria monocytogenes in fresh vegetables.

Two biopreservation approaches for fresh lettuce, rocket salad, parsley and spinach were studied. The potential of Pediococcus pentosaceus DT016, as a protective culture, to suppress Listeria monocytogenes in vegetables during storage was evaluated. The pathogen numbers in the vegetables inoculated with P. pentosaceus DT016 were significantly (p < 0.01) lower throughout the storage period and, at the last storage day, a minimum difference of 1.4 log CFU/g was reported when compared with the vegetables without the protective culture. Moreover, by using two levels of L. monocytogenes (about 6 and 4 log CFU/g), it was observed that the antagonist effect of P. pentosaceus was higher for the lower pathogen numbers. The second approach evaluated a pediocin DT016 solution to inactivate and control L. monocytogenes proliferation. The pathogen load was studied after washing with: water, chlorine and the pediocin solution and along storage at 4  °C. Comparing the various washing solutions, the vegetables washed with pediocin presented significantly (p < 0.01) lower pathogen numbers throughout storage, by a minimum of 3.2 and 2.7 log CFU/g, than in vegetables washed with water and chlorine, respectively. The proposed methodologies are promising alternatives to maintain the safety of fresh vegetables during extended storage at refrigeration temperature.

Non-thermal approach to Listeria monocytogenes inactivation in milk: The combined effect of high pressure, pediocin PA-1 and bacteriophage P100.

Non-thermal food processing and replacement of chemical additives by natural antimicrobials are promising trends in the food industry. The objective of the present work was to evaluate the effect of a process which combines mild high hydrostatic pressure - HHP (200 and 300 MPa, 5 min, 10 °C), phage Listex™ P100 and the bacteriocin pediocin PA-1 as a new non-thermal process for destruction of Listeria monocytogenes (104 CFU mL-1 or 107 CFU mL-1) in milk. For inoculum levels of 104 CFU mL-1, HHP combined with phage P100 eliminated L. monocytogenes immediately after pressurization. When L. monocytogenes was inoculated at levels of 107 CFU mL-1, a synergistic effect between phage P100, pediocin PA-1 and HHP (300 MPa) on the inactivation of L. monocytogenes was observed during storage of milk at 4 °C. For non-pressure treated samples inoculated with phage or pediocin or both, L. monocytogenes counts decreased immediately after biocontrol application, but regrowth was observed in a few samples during storage. Phage particles were stable during refrigerated storage for seven days while pediocin PA-1 remained stable only during three days. Further studies will have to be performed to validate the findings of this work in specific applications (e.g. production of raw milk cheese).

Stability of sunflower and rapeseed oil-in-water emulsions supplemented with ethanol-treated rapeseed meal protein isolate.

A protein isolate (ERPI) was prepared from ethanol-treated rapeseed meal and used as a stabilizing agent in sunflower and rapeseed oil-in-water emulsions. The aim of the current study was to explore the influence of protein and oil concentrations on initial stability of sunflower and rapeseed oil-in-water emulsions by evaluating Gibbs free energy (ΔG) and particle size distribution. The 7-day dynamics of emulsion stability was investigated by turbidity measurement as well. A 32 factorial design was applied to assess the significance of oil (5%, 10% and 15% w/w) and ERPI protein (0.25%, 0.5% and 1.0% w/w) addition on stability of the emulsions. The results demonstrated that the increase of oil concentrations from 5 to 15% positively influenced the initial stability of sunflower and rapeseed oil-in-water emulsions. In both oil types, ERPI protein supplementation at all levels resulted in significant differences in the stability of 5% and 10% oil emulsions but did not alter the initial stability of the emulsions prepared with either 15% sunflower or rapeseed oil. With a few exceptions, there was a good agreement between Gibbs free energy data and microstructural profiles of the emulsions. Overall, emulsions with all sunflower oil concentrations and 1.0% ERPI protein exhibited better initial and a 7-day stability dynamics compared to all rapeseed oil-based emulsions. The study demonstrated the potential of ethanol-treated rapeseed meal protein isolate to serve as an emulsifying agent in sunflower and rapeseed oil containing emulsions.

The protective effect of food matrices on Listeria lytic bacteriophage P100 application towards high pressure processing.

The application of lytic phages as biocontrol agents is emerging as a promising strategy towards elimination or reduction of foodborne pathogens in a variety of food products. This technology is particularly advantageous for minimally processed and ready-to-eat (RTE) foods. In this study, the potential use of Listex™ P100 combined with high hydrostatic pressure (HPP), to enhance the control of Listeria monocytogenes in food was evaluated. For that, the effect of three pressures (200, 300 or 400 MPa; 5 min, 10 °C) on phage P100 stability was tested when inoculated in six different matrices: phosphate buffered saline (PBS, pH 7.4); apple juice (pH 3.41); orange/carrot nectar (pH 3.54); UHT whole milk (pH 6.73); and, two traditional Portuguese fermented products, "Serra da Estrela" cheese (pH 5.66) and "Alheira", a meat sausage (pH 6.07). The results showed that treatment at 400 MPa reduced phage titres to below the detection level in all matrices, whereas at milder pressures the survival of the phage was matrix dependent. "Alheira", "Serra da Estrela" cheese and UHT whole milk were shown to be baroprotective matrices that support phage P100 application in HHP up to 300 MPa; however, an accentuated phage inactivation was observed in apple and orange/carrot nectar, which may be related to the acidic pH values of these matrices. The initial phage load did not affect the inactivation rate during HHP processing (300 MPa, 5 min, 10 °C) in PBS, cheese, sausage or milk matrices, and the phage titres were stable in these matrices during storage at 4 °C for 28 days for milk and 60 days for "Alheira" and "Serra da Estrela" cheese. In addition, a baroprotective effect on phage stability was observed when PBS was supplemented with reducing sugars, dextrin, casein, and tween 80. In conclusion, at mild HHP treatment, phage P100 remained active in specific matrices and seems to present potential to be added in non-thermal inactivation of L. monocytogenes.

Evaluation of antibiotic resistance patterns of food and clinical Listeria monocytogenes isolates in Portugal.

The aim of this study was to characterize a broad collection of isolates of Listeria monocytogenes, of different serotypes, recovered in Portugal between 2003 and 2007 from foods (n=353) and from clinical cases of human listeriosis (n=95), in terms of antimicrobial susceptibility. All the isolates were susceptible to ampicillin, the preferred agent to treat listeriosis. Resistances to nitrofurantoin (n=99), to ciprofloxacin (n=18), to erythromycin (n=10), to tetracycline (n=2), to gentamicin (n=1) and to rifampicin (n=1) were observed. One hundred (28.3%) and 20 (21.0%) food and clinical isolates, respectively, were resistant to at least one antibiotic. Eight isolates (1.8%) were resistant to two or more antimicrobials of different classes, and all were collected from foods. Serogroup IVb included the highest percentage of isolates resistant to erythromycin. The highest percentages of isolates resistant to nitrofurantoin were of serogroup IVb and IIc. It was demonstrated that the incidence of antibiotic-resistant isolates of L. monocytogenes, during the period 2003 to 2007, was low in Portugal but still higher than that observed in other countries. Given the increasing population at greater risk of listeriosis, namely, the elderly, the high mortality rate of the infection and the detection of resistant isolates, monitoring for antibiotic resistance in strains of L. monocytogenes on a large scale, and assessing the risk of infection by these strains, is highly recommended.

Kinetics of ethylene oxide desorption from sterilized materials.

Ethylene oxide gas is commonly used to sterilize medical devices, and concerns about using this agent on biological systems are well-established. Medical devices sterilized by ethylene oxide must be properly aerated to remove residual gas and by-products. In this work, kinetics of ethylene oxide desorption from different sterilized materials were studied in a range of aeration temperatures. The experimental data were well-described by a Fickian diffusion mass transfer behavior, and diffusivities were estimated for two textile and two polymeric materials within the temperature range of 1.5 to 59.0 degrees C. The results will allow predictions of ethylene oxide desorption, which is a key step for the design of sterilization/aeration processes, contributing to an efficient removal of residual ethylene oxide content.

Comparison of ubiquitous antibiotic-resistant Enterobacteriaceae populations isolated from wastewaters, surface waters and drinking waters.

This study aimed at assessing the role of ubiquitous (non-Escherichia coli) Enterobacteriaceae in the dissemination of antimicrobial resistance through the urban water cycle. Enterobacteriaceae isolated from a municipal wastewater treatment plant (111 isolates), urban water streams (33 isolates) and drinking water (123 isolates) were compared in terms of: (i) genera distribution, (ii) resistance to 12 antibiotics, and (iii) class 1 and class 2 integrons. The predominant bacterial genera were the same in the different types of water, although with a distinct pattern of species. The most prevalent resistance phenotypes were observed for amoxicillin, ticarcillin, cephalothin and sulphamethoxazole (24-59% in the three types of water). No resistance against ceftazidime or meropenem was observed. Resistance to cephalothin, amoxicillin and sulphamethoxazole was significantly more prevalent in drinking water, water streams and wastewater, respectively, than in the other types of water. It was possible to recognize antibiotic-resistance associations, namely for the pairs streptomycin-tetracycline (positive) and ticarcillin-cephalotin (negative). Class 1 and/or class 2 integrons with similar gene cassettes were detected in the three types of water. This study demonstrated that Enterobacteriaceae are important vehicles of antibiotic resistance, namely in drinking water.

Modelling growth of, and removal of Zn and Hg by a wild microalgal consortium.

Microorganisms isolated from sites contaminated with heavy metals usually possess a higher removal capacity than strains from regular cultures. Heavy metal-containing soil samples from an industrial dumpsite in Northern Portugal were accordingly collected; following enrichment under metal stress, a consortium of wild microalgae was obtained. Their ability to grow in the presence of, and their capacity to recover heavy metals was comprehensively studied; the datasets thus generated were fitted to by a combined model of biomass growth and metal uptake, derived from first principles. After exposure to 15 and 25 mg/L Zn(2+) for 6 days, the microalgal consortium reached similar, or higher cell density than the control; however, under 50 and 65 mg/L Zn(2+), 71% to 84% inhibition was observed. Growth in the presence of Hg(2+) was significantly inhibited, even at a concentration as low as 25 µg/L, and 90% inhibition was observed above 100 µg/L. The maximum amount of Zn(2+) removed was 21.3 mg/L, upon exposure to 25 mg/L for 6 day, whereas the maximum removal of Hg(2+) was 335 µg/L, upon 6 day in the presence of 350 µg/L. The aforementioned mechanistic model was built upon Monod assumptions (including heavy metal inhibition), coupled with Leudeking-Piret relationships between the rates of biomass growth and metal removal. The overall fits were good under all experimental conditions tested, thus conveying a useful tool for rational optimisation of microalga-mediated bioremediation.

Comparison of recovery methods for the enumeration of injured Listeria innocua cells under isothermal and non-isothermal treatments.

This study compares the feature of different media with the combination of selective with non-selective media in a TAL method for recovery of Listeria innocua cells exposed to thermal treatments. Experiments were conducted in broth at constant temperature (52.5 and 65.0 °C) and pH (4.5 and 7.5) conditions, using NaCl or glycerol to adjust water activity to 0.95. Four different media were used in bacterial cell enumeration: (i) a non-selective medium - TSAYE, (ii) two selective media - TSAYE + 5%NaCl and Palcam Agar and (iii) TAL medium (consisting of a layer of Palcam Agar overlaid with one of TSAYE). Two food products were used as case studies aiming at comparison of results obtained on selective and TAL media enumeration. Parsley samples were inoculated with L. innocua and subjected to posterior thermal treatments both under isothermal (52.5, 60.0 and 65.0 °C) and non-isothermal (heating rate of 1.8 °C/min from 20.0 to 65.0 °C) conditions. The recovery capability of TAL method was also studied when a pre-cooked frozen food (i.e. meat pockets) was fried (oil temperature of ∼180 °C). TAL method proved to be better than Palcam Agar in terms of capability to recover injured cells and was effective in L. innocua enumeration when non-sterile samples were analysed.

Survival of clinical and food isolates of Listeria monocytogenes through simulated gastrointestinal tract conditions.

Twenty-seven strains of Listeria monocytogenes previously isolated from food (n = 16) and human patients of listeriosis (n = 11) were characterized and compared based on their ability to survive through the simulated gastrointestinal tract conditions. Cells were exposed (60 or 120 min) to low pH in the presence of pepsin, to simulate the digestion in the stomach, and subsequently to bile salts to simulate the digestion in the small intestine (60 or 120 min). Their survival was shown to be origin- (food and clinical) and strain dependent (p < 0.001) and also significantly dependent on the imposed simulated gastric conditions (long vs. quick exposure) (p < 0.001). In comparison to the food isolates, the clinical strains were in general more resistant and survived better to the two challenges imposed. Some of the tested strains, after the exposure to low pH in the presence of pepsin, became injured and subsequently more susceptible to the bile salts challenge. It was demonstrated that one of the most important natural barriers against foodborne pathogens might not be effective since it was shown that L. monocytogenes isolates that survived through the pH challenge were also able to survive the subsequent challenge to bile salts.

Influence of pH, type of acid and recovery media on the thermal inactivation of Listeria innocua.

Acidification of foods with organic acids, either by fermentation or by intentional addition, is an important and common mechanism for controlling foodborne pathogens in a diversity of food products. The objective of this work was to study thermal inactivation of Listeria innocua, an acid tolerant microorganism, at 52.5, 60.0 and 65.0 degrees C, at different pH values (4.5, 6.0 and 7.5), using three types of acid (lactic, acetic and hydrochloric) and three different plating media (Tryptic Soy Agar with 0.6% yeast extract-TSAYE; TSAYE plus 5% NaCl-TSAYE+5%NaCl; and Palcam Agar with selective supplement-Palcam Agar), according to a 3(4) factorial experimental design. Survival data experimentally obtained were fitted with a Gompertz-inspired model and kinetic parameters (shoulder, maximum inactivation rate-k(max), and tail) were estimated for all conditions considered. The influence of temperature, pH, type of acid and enumeration media on kinetic parameters was assessed. Results showed that, with the exception of the type of acid, all the remaining factors and their combinations significantly affected the shoulder period and k(max). In relation to tail, temperature and recovery media were the affectable factors. It was concluded that the survival of this bacteria is higher when combining low temperature with neutral pH, and when TSAYE is the enumeration medium. Bigelow-inspired models were successfully developed and describe accurately the temperature and pH effects on the kinetic parameters.