M. Walkling-Ribeiro

Inactivation of Escherichia coli in a Tropical Fruit Smoothie by a Combination of Heat and Pulsed Electric Fields

Moderate heat in combination with pulsed electric fields (PEF) was investigated as a potential alternative to thermal pasteurization of a tropical fruit smoothie based on pineapple, banana, and coconut milk, inoculated with Escherichia coli K12. The smoothie was heated from 25 degrees C to either 45 or 55 degrees C over 60 s and subsequently cooled to 10 degrees C. PEF was applied at electric field strengths of 24 and 34 kV/cm with specific energy inputs of 350, 500, and 650 kJ/L. Both processing technologies were combined using heat (45 or 55 degrees C) and the most effective set of PEF conditions. Bacterial inactivation was estimated on standard and NaCl-supplemented tryptone soy agar (TSA) to enumerate sublethally injured cells. By increasing the temperature from 45 to 55 degrees C, a higher reduction in E. coli numbers (1 compared with 1.7 log(10) colony forming units {CFU} per milliliter, P < 0.05) was achieved. Similarly, as the field strength was increased during stand-alone PEF treatment from 24 to 34 kV/cm, a greater number of E. coli cells were inactivated (2.8 compared with 4.2 log(10) CFU/mL, P < 0.05). An increase in heating temperature from 45 to 55 degrees C during a combined heat/PEF hurdle approach induced a higher inactivation (5.1 compared with 6.9 log(10) CFU/mL, respectively [P < 0.05]) with the latter value comparable to the bacterial reduction of 6.3 log(10) CFU/mL (P> or = 0.05) achieved by thermal pasteurization (72 degrees C, 15 s). A reversed hurdle processing sequence did not affect bacterial inactivation (P> or = 0.05). No differences were observed (P> or = 0.05) between the bacterial counts estimated on nonselective and selective TSA, suggesting that sublethal cell injury did not occur during single PEF treatments or combined heat/PEF treatments.

Antimicrobial effect and shelf‐life extension by combined thermal and pulsed electric field treatment of milk

Aims:  The impact of a combined hurdle treatment of heat and pulsed electric fields (PEF) was studied on native microbiota used for the inoculation of low‐fat ultra‐high temperature (UHT) milk and whole raw milk. Microbiological shelf‐life of the latter following hurdle treatment or thermal pasteurization was also investigated.

Factors affecting the inactivation of the natural microbiota of milk processed by pulsed electric fields and cross-flow microfiltration

Prior to processing milk and cream were standardised and homogenised. Skim milk was cross-flow microfiltered (CFMF) prior to treatment with pulsed electric fields (PEF) or high temperature short time (HTST) pasteurization. The effect of temperature of the skim milk and product composition on the efficacy of PEF treatment was determined. The electrical conductivity of the product was related to fat and solids content and increased 5% for every g/kg increase of solids and decreased by nearly 0·7% for every g/kg increase of fat. From the three microbial groups analyzed (mesophilic, coliform, and psychrotroph) in milks differences (P<0·05) in the inactivation of mesophilic microorganisms were observed between the counts following PEF treatment, while HTST pasteurization resulted in higher reductions in all different counts than those obtained after PEF. Increasing the skim milk temperature prior to PEF treatment to about 34°C showed equivalent reductions in microbial counts to skim milk treated at 6°C in half the time. The reductions achieved by a combination of CFMF and PEF treatments were comparable to those achieved when CFMF was combined with HTST pasteurization. A higher reduction in coliform counts was observed in homogenised products subjected to PEF than in products that were only standardised for fat content.

Change in Color and Volatile Composition of Skim Milk Processed with Pulsed Electric Field and Microfiltration Treatments or Heat Pasteurization

Non-thermal processing methods, such as pulsed electric field (PEF) and tangential-flow microfiltration (TFMF), are emerging processing technologies that can minimize the deleterious effects of high temperature short time (HTST) pasteurization on quality attributes of skim milk. The present study investigates the impact of PEF and TFMF, alone or in combination, on color and volatile compounds in skim milk. PEF was applied at 28 or 40 kV/cm for 1122 to 2805 µs, while microfiltration (MF) was conducted using membranes with three pore sizes (lab-scale 0.65 and 1.2 µm TFMF, and pilot-scale 1.4 µm MF). HTST control treatments were applied at 75 or 95 °C for 20 and 45 s, respectively. Noticeable color changes were observed with the 0.65 µm TFMF treatment. No significant color changes were observed in PEF-treated, 1.2 µm TFMF-treated, HTST-treated, and 1.4 µm MF-treated skim milk (p ≥ 0.05) but the total color difference indicated better color retention with non-thermal preservation. The latter did not affect raw skim milk volatiles significantly after single or combined processing (p ≥ 0.05), but HTST caused considerable changes in their composition, including ketones, free fatty acids, hydrocarbons, and sulfur compounds (p < 0.05). The findings indicate that for the particular thermal and non-thermal treatments selected for this study, better retention of skim milk color and flavor components were obtained for the non-thermal treatments.

Structural changes induced by high-pressure processing in micellar casein and milk protein concentrates

Reconstituted micellar casein concentrates and milk protein concentrates of 2.5 and 10% (wt/vol) protein concentration were subjected to high-pressure processing at pressures from 150 to 450 MPa, for 15 min, at ambient temperature. The structural changes induced in milk proteins by high-pressure processing were investigated using a range of physical, physicochemical, and chemical methods, including dynamic light scattering, rheology, mid-infrared spectroscopy, scanning electron microscopy, proteomics, and soluble mineral analyses. The experimental data clearly indicate pressure-induced changes of casein micelles, as well as denaturation of serum proteins. Calcium-binding αS1- and αS2-casein levels increased in the soluble phase after all pressure treatments. Pressurization up to 350 MPa also increased levels of soluble calcium and phosphorus, in all samples and concentrations, whereas treatment at 450 MPa reduced the levels of soluble Ca and P. Experimental data suggest dissociation of calcium phosphate and subsequent casein micelle destabilization as a result of pressure treatment. Treatment of 10% micellar casein concentrate and 10% milk protein concentrate samples at 450 MPa resulted in weak, physical gels, which featured aggregates of uniformly distributed, casein substructures of 15 to 20 nm in diameter. Serum proteins were significantly denatured by pressures above 250 MPa. These results provide information on pressure-induced changes in high-concentration protein systems, and may inform the development on new milk protein-based foods with novel textures and potentially high nutritional quality, of particular interest being the soft gel structures formed at high pressure levels.

Effect of heat-assisted pulsed electric fields and bacteriophage on enterohemorrhagic Escherichia coli O157:H7.

Pulsed electric fields (PEF), heat‐assisted PEF (H‐PEF), and virulent bacteriophage (VP) are non‐thermal techniques for pathogen inactivation in liquids that were investigated individually, and in combination (PEF/VP, H‐PEF/VP) to control enterohemorrhagic Escherichia coli (EHEC) O157:H7 in Luria‐Bertani broth (LBB) and Ringers solution (RS). Treated cells were subsequently incubated at refrigeration (4°C) and temperature‐abuse conditions (12°C) for 5 days. When EHEC cells grown in LBB were subjected to non‐thermal processing and subsequently stored at 12°C for 5 days, reductions in count of between 0.1 and 0.6 log cycles were observed and following storage at 4°C the decrease in counts varied between 0.2 and 1.1 log10. For bacteria cells suspended in RS values ranged from 0.1 to ≥3.9 log cycles at both storage temperatures. The most effective treatments were H‐PEF and H‐PEF/VP, both producing a >3.4 log cycle reduction of cells suspended in non‐nutrient RS. Analysis of EHEC recovery on selective and non‐selective media indicated no occurrence of sub‐lethal damage for VP, PEF/VP, and H‐PEF/VP‐treated cells. The findings indicate that combining PEF and lytic phage may represent a suitable alternative to conventional fluid decontamination following further process optimization.

Pulsed electric field processing preserves the antiproliferative activity of the milk fat globule membrane on colon carcinoma cells

The present work evaluated the effect of processing on the antiproliferative activities of milk fat globule membrane (MFGM) extracts. The antiproliferative activity on human adenocarcinoma HT-29 cells of untreated MFGM extracts were compared with those extracted from pasteurized cream, thermally treated cream, or cream subjected to pulsed electrical field (PEF) processing. The PEF with a 37 kV/cm field strength applied for 1,705μs at 50 and 65°C was applied to untreated cream collected from a local dairy. Heating at 50 or 65°C for 3min (the passage time in the PEF chamber) was also tested to evaluate the heating effect during PEF treatments. The MFGM extracted from pasteurized cream did not show an antiproliferative activity. On the other hand, isolates from PEF-treated cream showed activity similar to that of untreated samples. It was also shown that PEF induced interactions between β-lactoglobulin and MFGM proteins at 65°C, whereas the phospholipid composition remained unaltered. This work demonstrates the potential of PEF not only a means to produce a microbiologically safe product, but also as a process preserving the biofunctionality of the MFGM.

Chapter 7 – Pulsed Electric Field Processing of Liquid Foods and Beverages

Presently, most liquid foods are preserved commercially by ultra-high temperature or high temperature short time processes. Although heating inactivates enzymes and microorganisms, the organoleptic and nutritional properties of the food suffer because of protein denaturation and the loss of vitamins and volatile flavors. Thus, extending the shelf life of food by heat treatment is not only energy intensive, but, in many cases, adversely affects the flavor, chemical composition, and nutritional quality of the treated food. There is a great need for a nonthermal method for inactivating microorganisms that is economical, compact, energy efficient, safe, socially and environmentally acceptable, and which does not adversely affect nutrition, texture, and flavor of the treated food. Consumers are also increasingly demanding high-quality, minimally processed foods. Overall, the pulsed electric field process has great potential for use as a preservation treatment for diverse liquid foods and, although North America and Europe are still the global regions that have the highest research output regarding pulsed electric field, they are not the only driving forces for pulsed electric field research anymore.

Microbial decontamination of milk and dairy products

Abstract: Despite advances in dairy production and processing methods, outbreaks of illness associated with milk and milk products continue to occur. At the same time there has been a resurgence of interest by consumers in organic, ‘slow’ and ‘raw’ foods. To offset issues related to food safety and to comply with the demand for these fresh foods, there is an urgent need to develop novel processing concepts that provide effective public health protection with minimal treatments to retain nutrient and flavour characteristics of the product. In the first part of this chapter all milk and dairy pathogens of relevance are discussed with particular focus on their relative risk ranking. The second part of the chapter reviews the features and potential of conventional and most promising emerging preservation techniques for pathogen mitigation in milk and dairy products, highlighting the advantages of combined decontamination strategies and indicating hurdle processing as the most efficacious approach.