Antimicrobial plastic film: Physico-chemical characterization and nisin desorption modeling

Antimicrobial active films represent an innovative concept in food packaging, developed to answer to consumer's expectation for better microbiological safety. In this study, the growth of pathogenic micro-organisms on the surface of food is proposed to be controlled by coating, on the surface of polyethylene/polyamide/polyethylene film (PE/PA/PE), a film-forming solution containing Nisaplin, a commercial form of bacteriocin produced by Lactococcus lactis subsp. lactis: nisin. The bioactivity of these multi-layer films coated with Nisaplin loaded HydroxyPropylMethylCellulose film is based on the release of this antimicrobial molecule towards a food simulant. Nisin mass transfer was studied and modeled, for different operating conditions, generally encountered in food products. pH didn't seem to interfere with nisin release kinetics, while the variation of NaCl concentration between 0.8% and 3.2% decreased the desorption coefficient (k_d) by 18% and the temperature increase from 10 °C to 28 °C resulted in an increase of k_d from 1.78 × 10^? 2 m s^? 1 to 2.10 × 10^? 2 m s^? 1. Coating of PE/PA/PE film with this antimicrobial layer induced little mechanical properties modifications without compromising industrial applications. Water barrier capacity was not altered.Industrial relevanceThis paper concerns active packaging, considered as a new approach to preserve food shelf life. Active packaging is a real gain for plastic and Food industrials. Coating was used to obtain antimicrobial packaging. The impact of coating on film characteristics is investigated.Also, antimicrobial agent desorption is determined during storage conditions.     

Time–temperature study of the kinetics of migration of DPBD from plastics into chocolate,chocolate spread and margarine

Migration of low molecular weight substances into foodstuffs is a subject of increasing interest and an important aspect of food packaging because of the possible hazardous effects on human health.The migration of a model substance (diphenylbutadiene) from low-density polyethylene (LDPE) was studied in foodstuffs with high fat contents: chocolate, chocolate spread and margarines (containing 61% and 80% fat).A simplifying mathematical model based on Fick’s diffusion equation for mass transport processes from plastics was used to derive effective diffusion coefficients which take also kinetic effects in the foods into account and to determine partition coefficients between plastic and food. With this model migration levels obtainable under other storage conditions can be predicted. The effective diffusion coefficients for both margarines stored at 5 °C (3.0–4.2 × 10⁻¹⁰ cm² s⁻¹) and at 25 °C (3.7–5.1 × 10⁻⁹ cm²  s⁻¹) were similar to each other, lower than for chocolate spread stored at 5 °C (9.1 × 10⁻¹⁰ cm² s⁻¹) and higher than the diffusion coefficient for chocolate stored at 25 °C (2.9 × 10⁻¹⁰ cm² s⁻¹). Good agreement was found between the experimental and the estimated data, allowing validation of this model for predicting diffusion processes in foodstuffs with high fat contents.     

Migration of α-tocopherol from LDPE films to corn oil and its effect on the oxidative stability

The migration of α-tocopherol (α-T) from low density polyethylene (LDPE) films, added with 20 (film A) and 40 mg g^?1 (film B) to corn oil for 12 weeks at 5, 20 and 30 °C was determined. A LDPE film added with no α-T was used as control (film C). Diffusion coefficient (D) values for the film A system were 1.4 × 10^?11, 7.1 × 10^?11 and 30.3 × 10^?11 cm² s^?1 at 5, 20 and 30 °C, respectively. Meanwhile, D values for the film B system were 1.3 × 10^?11, 9.6 × 10^?11 and 51.1 × 10^?11 cm² s^?1 at the same temperatures. The activation energy (E_a) for the diffusion of α-T was 126.5 (film A) and 105.9 kJ mol^?1 (film B). The effect of the migration of α-T on the oxidative stability of corn oil was evaluated by monitoring hexanal content by solid phase micro-extraction (SPME) and gas chromatography. The hexanal content in the oil showed that both films added with α-T resulted suitable to maintain the oxidative stability of the oil for about 16 weeks at 30 °C, compared to 12 weeks for the oil in contact with the film C.     

Innovative multilayer antimicrobial films made with Nisaplin® or nisin and cellulosic ethers: Physico-chemical characterization,bioactivity and nisin desorption kinetics

In this study, ethylcellulose/hydroxypropylmethylcellulose/ethylcellulose (EC/HPMC/EC) three-layer films including Nisaplin® or nisin and lecithins were formulated. Lecithins were used as plasticizers to ensure cohesion between hydrophobic ethylcellulose and hydrophilic HPMC layers. It was observed that the introduction of pure nisin or its non-pure commercial form Nisaplin® into films didn't significantly alter their mechanical and optical properties. Additionally, these nisin or Nisaplin-loaded multilayer films showed significant antimicrobial activity. The comparison of inhibition diameters obtained with EC/HPMC film used as control and EC/HPMC/EC films demonstrated that the three-layer films delayed nisin desorption. This was confirmed by the kinetics of nisin release in a (0.8% w/v) NaCl solution at 28 °C: nisin from two-layer EC/HPMC films totally desorbed within 0.5 h, while the three-layer films allowed to expand nisin release time over 20 h. The ratio of nisin desorption coefficients (k_d): k_{d (EC/HPMC)}/k_{d (EC/HPMC/EC)} was determined after desorption modelling, and was found to be up to 118, proving that multilayer films with hydrophobic layers could be a potential way to control nisin release from antimicrobial bio-packagings.Industrial relevanceThis paper concerns active packaging, considered as a new approach to preserve food shelf life. Active packaging is a real gain for plastic and food industrials. Coating was used to obtain antimicrobial packaging. The impact of incorporating the antimicrobial agent in multilayer films on the release kinetics is investigated.     

Effect of temperature,pH and film thickness on nisin release from antimicrobial whey protein isolate edible films

BACKGROUND: An active packaging film based on whey protein isolate (WPI) was developed by incorporating nisin to promote microbial food safety. The effect of temperature and pH on the release of nisin from edible films of different thickness was investigated. The film mechanical properties and inhibitory effect were also evaluated. RESULTS: Nisin release was significantly favoured by low pH, with the highest release after 24 h (1325 IU), which was not significantly affected by temperature (5 or 10 °C). Thickness significantly affected film elongation, with thicker films showing the highest elongation (54.3 ± 2.7%). Water vapour permeability (0.15 ± 0.4 g mm m^?2 kPa^?1 h^?1) and elastic modulus were not significantly affected by thickness. The highest nisin effective diffusivity (5.88 × 10^?14 m² s^?1) was obtained using a solution at pH 4, 112 µm film thickness and a temperature of 5 °C. More than four log cycles of Brochotrix thermosphacta were reduced from the surface of a ham sample after 8 days of incubation at 4 °C by the active WPI film containing 473 IU cm^?2 nisin. CONCLUSION: Nisin diffusivity from WPI edible films was favoured at lower pH and film thickness. This active packaging film may be used to preserve the quality and safety of meat products. Copyright © 2009 Society of Chemical Industry     

Evaluation of biodegradable film packaging to improve the shelf-life of Boletus edulis wild edible mushrooms

The objective of this work was to evaluate the effect of poly(lactic acid) (PLA) based biodegradable film packaging combining 0.5% nisin antimicrobial polypeptide on the physicochemical and microbial quality of Boletus edulis wild edible mushrooms stored at 4 ± 1 °C. The experiment was set up by packaging mushrooms with extruded PLA films containing 0, 7.5, and 15 wt.% triethyl citrate plasticizer. The low-density polyethylene (LDPE) film was used as the control. Mushrooms stored in PLA films containing 7.5 and 15 wt.% plasticizer provided better retention of quality characteristics and received higher sensory ratings compared to mushrooms stored in pure PLA film and LDPE film. Samples with these two treatments underwent minimal changes in texture, PPO activity, total bacteria count, and sensory attributes. Results suggest that nisin in combination with plasticized PLA film has the potential to maintain B. edulis wild edible mushroom quality and extend its postharvest life to 18 days.Industrial relevanceB. edulis is one of the most commercialized mushrooms worldwide. However, as with all fresh mushrooms, there are severe preservation problems. Extruded PLA films containing triethyl citrate plasticizer plus antimicrobial agent nisin proved to be a suitable technology for mushroom conservation. This material exhibits an environmental-friendliness potential and a high versatility in food packaging.     

Combinations of nisin with organic acids or salts to control Listeria monocytogenes on sliced pork bologna stored at 4°C in vacuum packages

This study evaluated dipping solutions of nisin with or without organic acids or salts, as inhibitors of Listeria monocytogenes introduced on sliced cooked pork bologna before vacuum packaging and storage at 4°C for 120 days. Inoculated (10²–10³ cfu/cm²) slices were immersed in nisin (5000 IU/ml), or in lactic or acetic acid (1, 3, 5 g/100 ml), sodium acetate or diacetate (3, 5 g/100 ml), and potassium benzoate or sorbate (3 g/100 ml), each combined with nisin. Additional slices were immersed in nisin, inoculated and then immersed in acid or salt solutions without nisin. Nisin reduced L. monocytogenes by 1.0–1.5 log cfu/cm² at treatment (day-0) followed by a listeriostatic effect for 10 days. Thereafter, however, the pathogen multiplied in treatments without acid or salts, with growth being faster on slices immersed in nisin after as compared to before inoculation. Nisin in combination with 3 or 5 g/100 ml acetic acid or sodium diacetate or 3 g/100 ml potassium benzoate, applied individually or as mixtures, did not permit growth before day-90. Other treatments were of variable and lesser effectiveness (20–70 days), whereas in untreated or water-treated (control) bologna L. monocytogenes increased at 6–7 log cfu/cm² at day-20. Based on the antilisterial efficacy and effects of treatments on product pH, nisin with 3 g/100 ml sodium diacetate may be the most promising combination in dipping solutions to control L. monocytogenes on sliced cured pork bologna.     

Rapid purification of nisin Z using specific monoclonal antibody-coated magnetic beads

A rapid method is described for the isolation and purification of nisin Z from Lactococcus lactis subsp. lactis biovar. diacetylactis UL719 culture supernatant using magnetic beads. Anti-nisin Z monoclonal antibodies were covalently bound to EDC-activated magnetic beads and the complex was incubated overnight at 4°C with the culture supernatant. Bound nisin Z was then eluted with urea 6 mformic acid 0.1 m mixture. Using this procedure, a high yield of 61.5% was obtained with a nisin Z specific activity of 2.4×10⁴ IU mg⁻¹. Magnetic beads were re-used for two other purification cycles during which a significant decrease of purification yield from 61.5 to 26.7% was observed. The immunomagnetic purification strategy developed in this work was shown to be efficient and offers an alternative rapid procedure for production of high-purity food-grade nisin Z.     

Nano-TiO2 particles and high hydrostatic pressure treatment for improving functionality of polyvinyl alcohol and chitosan composite films and nano-TiO2 migration from film matrix in food simulants

This study investigated the impact of nano-TiO₂ and high hydrostatic pressure (HHP) treatment on microstructure, water vapor and gas barrier, antibacterial and mechanical properties of polyvinyl alcohol (PVA)–chitosan (CHI) biodegradable films and determined the migration behavior of TiO₂ nanoparticles from the films to food simulants. Apart from the effect of filler, TiO₂ nanoparticles also could improve the antibacterial activity of the films and play a role as a plasticizer in the films. HHP treatment promoted the interaction between PVA and chitosan molecules, resulting in the formation of more compacted network structures in PVA–CHI films. The migration of TiO₂ from the films was investigated in food simulants including distilled water, acetic acid, ethanol and olive oil, in which the trace amount of TiO₂ (< 4.20 × 10^− 3‰) was only detected in olive oil. HHP treatment at 200–400 MPa significantly reduced migration of TiO₂ nanoparticles from the films.Industrial relevanceResults from this study provide a new application direction of high hydrostatic pressure (HHP) in the field of food packaging materials for improving the functionality of materials. Due to the non-thermal characteristic, HHP in combination with nano-TiO₂ not only improved the mechanical and barrier properties of the biodegradable PVA–CHI composite films (polyvinyl alcohol and chitosan based materials), but also enhanced the antibacterial activity of the films. The HHP treated PVA–CHI–TiO₂ films are very stable in food simulants, such as olive oils. Therefore, the utilization of HHP and nano-TiO₂ is promising in the preparation of food packaging materials with desirable functionalities.     

Effects of treatment with nisin on the microbial flora and sensory properties of a Greek soft acid-curd cheese stored aerobically at 4 °C

The present study evaluated the use of nisin as an antimicrobial treatment for shelf-life extension of Galotyri, a Greek soft acid-curd cheese, stored aerobically under refrigeration for a period of 42 days. Three different treatments were tested: N0, control sample with no nisin added; N1, 50 IU g⁻¹ nisin; and N2, 150 IU g⁻¹ nisin, the latter two treatments added post-production to the Galotyri cheese. Of all microorganisms enumerated, lactobacilli, lactococci and yeasts were the groups that prevailed in cheese samples, irrespective of antimicrobial treatment. Based primarily on sensory evaluation (appearance and taste) and a microbiological acceptability limit for yeasts (5 log cfu g⁻¹), the use of nisin treatments extended the shelf-life of fresh Galotyri cheese stored at 4 °C by ca. 7 days (N1) and 21 days (N2) with cheese maintaining good sensory characteristics.     

Estimation of the diffusivities of sodium chloride,potassium sorbate and sodium bisulphite in mango slices processed by hurdle technology

The fractional amount of sodium chloride, potassium sorbate and sodium bisulphite were evaluated in mango slices immersed in limited volumes of syrup at 25, 50 and 70 °C. The syrup contained 250 g sucrose, 1.5 g sodium chloride, 0.5 g potassium sorbate and 0.25 g sodium bisulphate per kilogram of solution. The sodium chloride concentration in the syrup was confirmed with a flame photometer, and the concentrations of potassium sorbate and sodium bisulphite were determined using high-performance liquid chromatography (HPLC). Fick’s second law was used to calculate effective diffusion coefficients and to predict solute content in the mango slices. Diffusion coefficients were affected by temperature and were correlated by the Arrhenius equation. The experimental data fit the proposed mathematical model well, allowing prediction of the system’s behavior at different temperatures. The resultant diffusivities ranges were 2.63–3.54 × 10^?9 m²/s for sodium chloride, 3.88 × 10^?9–8.3 × 10^?10 m²/s for potassium sorbate and 1.83 × 10^?7–5.98 × 10^-8 m²/s for sodium bisulphite.     

Combined effect of γ-irradiation and osmotic pretreatment on mass transfer during dehydration

The combined effect of γ-irradiation (3.0–9.0 kGy) and osmotic pretreatment (10 and 50°B) on dehydration kinetics was studied. The exposure to irradiation pretreatment resulted in an increase in cell wall permeabilization, leading to softening of tissue, which in turn resulted in faster dehydration. The effective diffusion coefficient of water in case of potato during dehydration, considering Fickian diffusion, increased from 2.38 × 10⁻⁹ to 4.14 × 10⁻⁹ m²/s at 9.0 kGy, whereas the osmotic pretreatment resulted in lower dehydration rates due to infusion of solute from osmotic solution, which resulted in lower mass transfer rates. Even though the effective diffusion coefficients decreased from 2.38 × 10⁻⁹ to 1.55 × 10⁻⁹ m²/s (up to 50°B), the osmotic pretreatment improves nutritional, sensorial and functional properties of food without changing its integrity, apart from partial removal of water present in food. The combining of γ-irradiation treatment with osmotic pretreatment resulted in increased mass transfer rates during air dehydration, offering a feasible solution for satisfactorily enhancing the mass transfer rates.     

Diffusivity of potassium sorbate in κ-carrageenan based antimicrobial film

The diffusion of potassium sorbate incorporated into κ-carrageenan based antimicrobial film was measured at 5 s intervals using a diffusion cell. The kinetics of potassium sorbate release followed Fick's law of diffusion, as shown by diffusional exponent characteristic n-value and high correlation coefficients between experimental and theoretical data. The effects of solution pH at the film surface (3.8, 5.2 and 7.0) and temperature (5, 25 and 40 °C) on diffusion were investigated. Diffusion was found to be unaffected by adjacent solution pH in the range of values tested, but a decrease in temperature from 40 to 5 °C resulted in a reduction of diffusion coefficients from 4.24×10⁻¹³ to 1.29×10⁻¹³ m²/s at pH 5.2.     

Aerosol-assisted low pressure plasma deposition of antimicrobial hybrid organic-inorganic Cu-composite thin films for food packaging applications

An innovative low pressure plasma process for deposition of copper-containing hybrid organic-inorganic thin films was developed. The discharge was fed with an aerosol of an aqueous solution of a copper complex, i.e. bis(ethylenediamine)copper(II) hydroxide and argon. Polymeric films, incorporating inorganic Cu(I) and Cu(II) compounds, were obtained. Morphological and chemical characterizations of the coatings were carried out.Antimicrobial properties were assessed on two species of Pseudomonas spp. In vitro tests were carried out by contact of the optimized films, deposited on square polycarbonate samples (~ 4 cm²), with cell suspensions of 1 × 10⁴ CFU/mL for 18 h at 25 °C. It was demonstrated that, the hybrid organic-inorganic thin coatings have potential utilization as active packaging material, showing an antimicrobial effect of up to three orders of magnitude (final microbial concentration 10⁵ CFU/mL), compared to control polycarbonate (final microbial concentration 10⁸ CFU/mL).Industrial relevanceAn innovative low pressure plasma process for deposition of copper-based, hybrid organic-inorganic thin films was developed. The optimized thin coatings have potential industrial utilization as active packaging material, being very effective against pseudomonads. Viability of Pseudomonas was reduced by three orders of magnitude (from 10⁸ CFU/mL to 10⁵ CFU/mL) in the presence of developed films, thus suggesting further investigation of the technique under food packaging conditions.     

Effects of flaxseed oil concentration on the performance of a soy protein isolate-based emulsion-type film

Biodegradable edible films have the potential to either replace or reduce the amount of synthetic packaging utilized by the food industry. The overall goal of this research was to investigate the effect of flax seed oil concentration (1–10%) on the mechanical, moisture barrier and swelling properties of soy protein isolate (SPI) (5.0% w/w SPI, 40% w/w glycerol) emulsion-based films. Film forming solutions showed a bimodal oil droplet distribution with peak sizes occurring at < 10 and ~ 100 μm. As the oil content increased, the size distribution shifted towards smaller droplet sizes. An equal size ratio was noted at the 5.0% oil content level. All film forming solutions were pseudoplastic in nature, where viscosity increased from 18 to 58 mPa (at 1 s^− 1) as a function of oil content (3% to 10%). Tensile strength of formed films reached a maximum at 5.35 MPa at the 5% w/w oil level, whereas tensile elongation increased from 11.3% to 22.2% with increasing oil content. Puncture strength and deformation, as well as water vapour permeability was relatively independent of the oil content. Moisture content and swelling properties of formed films were found to both decrease from 22.8% to 18.7%, and from 3114% to 1209%, respectively as the oil content was raised from 1 to 10%, and films became darker, redder and more yellow in colour as the percentage of flax seed oil increased.     

Modeling sulfur dioxide uptake in dent corn during steeping

A mathematical model is employed to describe sulfur dioxide (SO₂) diffusion and reaction during steeping of dent corn. Experiments are performed to measure change of SO₂ content of grain during process. A computer-aided nonlinear optimization technique is used to estimate the effective diffusion coefficients and rate constants in the temperature range 25–55 °C. The effective diffusion coefficient for SO₂ varied between 2.27×10⁻¹¹ and 6.24×10⁻¹¹ m²/s and had an Arrhenius activation energy of 24.3 kJ/mol. The reaction rate of SO₂ in dent corn followed first-order kinetics, with rate constants in the range of 0.80×10⁻⁶–5.38×10⁻⁶ s⁻¹ and activation energy of 49.16 kJ/mol.     

Water vapor transport properties during staling of bread crumb and crust as affected by heating rate

In order to develop a mathematical model to simulate mass transfer occurring between the crumb and the crust during bread staling, water vapor sorption properties, i.e., moisture diffusivity, WVP and sorption of bread crumb and crust were investigated at 15 °C. Two types of bread baked with two heating rates (7.39 °C/min and 6.32 °C/min) were considered. Sorption and desorption isotherms were determined using Dynamic Vapor Sorption (DVS) and FF and GAB models were applied in the range of 0–0.95 a_w, to fit isotherm curves. Diffusivity was determined from sorption isotherms by using Fick's law and WVP was measured by two methods (gravimetric and from sorption data). Results exhibited maximum values of D_eff in the range of 0.1 and 0.14 g/g d.b. moisture contents. They varied between 0.88 × 10^? 10 and 0.92 × 10^? 10 m²/s for the crust and between 2.24 × 10^? 10 and 2.64 × 10^? 10 m²/s for the crumb, baked respectively at 220 °C and 240 °C. Results of WVP showed that the crust baked at 240 °C was significantly more permeable than the crust baked at 220 °C. This fact was attributed to the difference in porosity and the molecular structure due to heating effects. Also, the presence of steam in the oven atmosphere enhanced the development of higher porosity in the crust, leading to different structures and properties. Moreover, SEM images showed that starch granules were intact and less swelled in the upper crust when baking at 240 °C, resulting in higher WVP.     

Development and characterization of phosphatidylcholine nanovesicles containing the antimicrobial peptide nisin

Two methodologies were compared to encapsulate nisin in liposomes of partially purified soybean phosphatidylcholine: reversed-phase and hydration film. In the hydration film method, both probe-type and bath-type ultrasound were evaluated. The size of liposomes was evaluated by light scattering analysis and residual antimicrobial activities by agar diffusion assay using Listeria monocytogenes ATCC 7644 as indicator strain. The size of liposomes prepared by reversed-phase, hydration film using probe-type and bath-type ultrasound were 190, 181 and 148 nm with residual antimicrobial activities after encapsulation of 25%, 50% and 100%, respectively. The methodology of film hydration using bath-type ultrasound was chosen for assessment of its physicochemical characteristics. Nisin had entrapment efficiency of 94.12%. Measured Zeta potentials for unfiltered and filtered (0.22 μm) liposomes were ?55.8 and ?54.5 mV, respectively. The antimicrobial activity of free nisin, encapsulated nisin and filtered was evaluated for a period of 24 days. It was observed that the free nisin remained 100% of residual activity while the liposomes containing nisin were losing their antimicrobial activity over time reaching 25% residual activity after 10 days. The size (132–149 nm) and pH (4.5) remained constant over time. It was observed by microscopy that the liposomes maintained their spherical morphology. The stability observed by size and pH was not the same regarding antimicrobial activity and Zeta potential, indicating that the liposomes should be applied shortly after its preparation.     

Antimicrobial activity of nanoliposomes co-encapsulating nisin and garlic extract against Gram-positive and Gram-negative bacteria in milk

Nisin and garlic extract (GE) were co-encapsulated into phosphatidylcholine nanoliposomes. The mean diameter and zeta potential of the nanoparticles were 179 nm and − 27.7 mV, respectively, with an entrapment efficiency of about 82% and 90% for nisin and GE, respectively. The efficiency of free and encapsulated nisin-GE to control the development of Listeria monocytogenes, Salmonella Enteritidis, Escherichia coli and Staphylococcus aureus was assessed over time in whole milk at 37 °C. At such abuse temperature conditions, both free and liposomal nisin-GE resulted a difference of 1–4 log CFU/ml against the strains tested, when compared with free nisin and GE separately. A difference of 5–6 log CFU/ml in viable counts of Gram-positive strains and 3–4 log CFU/ml for Gram-negative bacteria was observed for treatments with nisin-GE when compared to the control. The effect of nisin-GE on L. monocytogenes was evaluated under refrigeration (7 ± 1 °C) for up to 25 days. Viable counts for treatments with free and encapsulated nisin-GE were 4–5 log CFU/ml lower than the values reached by the control. Liposome encapsulation of natural antimicrobials with synergistic effect may be important to overcome stability issues and undesirable interaction with food components. The results of this study indicated that nanoliposome-encapsulated nisin-GE has potential as an antimicrobial formulation for food use.Industrial relevanceThere is increased interest for minimally processed foods and natural additives, which agrees with the use of natural antimicrobials as food preservatives. Co-encapsulation of antimicrobials may extend the inhibitory spectrum and effectiveness in controlling food pathogens. The use of nanoliposomes for delivery of natural antimicrobials in dairy products represents an interesting alternative for controlled release of biopreservatives and improvement of food quality and shelf life.     

A risk characterization model of Salmonella Typhimurium in Irish fresh pork sausages

A second-order simulation model was built to estimate the risk of Salmonella Typhimurium associated with the consumption of Irish fresh pork sausages. To select appropriate hazard characterization models, an initial appraisal of the current dose-response models was conducted. The cooking modality of grilling was associated with a higher mean risk of infection per serving (1.399 × 10^? 6; 95% CI: 7.54 × 10^? 7–2.65 × 10^? 6) than frying (6.246 × 10^? 7, 95% CI: 2.78 × 10^? 7–1.17 × 10^? 6). When the risk was extrapolated over the consumption in a year period, the mean risk of infection increased considerably to 8.541 × 10^? 5 with an expected number of infections and illnesses of 184.3 (95% CI: 26–664) and 17 (95% CI: 2–63), respectively. Results highlighted the importance of consumer education, as scenario analysis predicted that, for the current level of Salmonella in pork sausage, decreasing the product's cold storage by approximately 8 h and cooking for an additional half minute can reduce the current risk level by ～ 50%.