High‐Pressure Thermal Sterilization: Food Safety and Food Quality of Baby Food Puree

The benefits that high‐pressure thermal sterilization offers as an emerging technology could be used to produce a better overall food quality. Due to shorter dwell times and lower thermal load applied to the product in comparison to the thermal retorting, lower numbers and quantities of unwanted food processing contaminants (FPCs), for example, furan, acrylamide, HMF, and MCPD‐esters could be formed. Two spore strains were used to test the technique; Geobacillus stearothermophilus and Bacillus amyloliquefaciens, over the temperature range 90 to 121 °C at 600 MPa. The treatments were carried out in baby food puree and ACES‐buffer. The treatments at 90 and 105 °C showed that G. stearothermophilus is more pressure‐sensitive than B. amyloliquefaciens. The formation of FPCs was monitored during the sterilization process and compared to the amounts found in retorted samples of the same food. The amounts of furan could be reduced between 81% to 96% in comparison to retorting for the tested temperature pressure combination even at sterilization conditions of F₀‐value in 7 min.     

Proteolytic and angiotensin‐converting enzyme‐inhibitory activities of selected probiotic bacteria

This study was carried out to examine the proteolytic and angiotensin‐converting enzyme (ACE‐I) activities of probiotic lactic acid bacteria (LAB) as influenced by the type of media, fermentation time, strain type and media supplementation with a proteolytic enzyme (Flavourzyme^®). Lactobacillus casei (Lc210), Bifidobacterium animalis ssp12 (Bb12), Lactobacillus delbrueckii subsp. bulgaricus (Lb11842) and Lactobacillus acidophilus (La2410) were grown in 12% of reconstituted skim milk (RSM) or 4% of whey protein concentrates (WPC‐35) with or without combination (0.14%) of Flavourzyme^® for 12 h at 37 °C. All the strains were able to grow in both media depending on type of strains used and fermentation time. All the strains showed higher proteolytic activity and produced more antihypersensitive peptides when grown in RSM medium at 12 h, when compared to WPC. Combination with Flavourzyme^® also increased LAB growth and proteolytic and ACE‐I activities. Of the four strains used, Bb12 and La2410 outperformed Lc210 and Lb11842. The highest ACE‐I activity and proteolytic activity were found in B. animalis ssp12 combined with Flavourzyme^®. Flavourzyme^® led to an increase in the production of bioactive peptides with ACE‐I activity during 12 h at 37 °C.     

Production and characterization of milk-clotting enzyme from <Emphasis Type="Italic">Bacillus amyloliquefaciens</Emphasis> JNU002 by submerged fermentation

Milk-clotting enzymes produced by microorganisms have been developed to replace calf rennet, yet the enzymatic level and the ratio of milk-clotting activity to proteolytic activity still need further improvement. This work described a strain Bacillus amyloliquefaciens JNU002 that screened from wheat bran that has a promising characterization. After optimization, B. amyloliquefaciens JNU002 showed a high milk-clotting activity (4969 SU/mL) and low proteolytic activity (4.02 U/mL) at 48 h with an inoculum size of 0.2% (v/v) at initial pH 6.0 in 15-L bioreactor. After purification, the purified enzyme gave a single protein band on SDS–PAGE, corresponding to 28 kDa. The purified enzyme showed a high ratio (2,575) of milk-clotting activity to proteolytic activity at 35 °C, and the ratio would be even higher (22,992) at 70 °C. The milk-clotting reaction and proteolytic reaction were prevented at 75 °C. This enzyme was stable at pH 4–6 and below 40 °C, and this was convenient for storage and transportation.     

High‐pressure processing of Manuka honey: brown pigment formation,improvement of antibacterial activity and hydroxymethylfurfural content

This work was undertaken to assess the effects of high‐pressure processing on brown pigment formation, antibacterial activity and hydroxymethylfurfural (HMF) content in Manuka honey. The honey was subjected to different pressures (200–600 MPa) at ambient temperatures (25–33 °C) and with moderate temperatures (53–74 °C) for holding times (10–30 min). The brown pigment formation in high‐pressure‐treated sample showed a significant increase (0.16–0.17, P < 0.05), whereas maximum percentage inhibition of Staphylococcus epidermidis (84.34 ± 7.62%) was achieved when honey was subjected to 600 MPa as compared to the control (64.15 ± 5.86%). The percentage inhibition in high‐pressure‐treated samples correlated linearly (r = 0.941) with brown pigment. No significant increase in HMF as affected by high pressure was found (P > 0.05). Thus, high‐pressure processing has no adverse effect on the honey quality because it can control HMF concentration during process. Increasing brown pigment formation is associated with the enhancement of antibacterial activity.     

Partial characterisation of enterocin KP produced by Enterococcus faecalis KP,a cheese isolate

Enterococcus faecalis KP, a bacteriocin‐producing strain, was identified using 16S rRNA gene sequence homology (99%). Enterocin KP from E. faecalis KP was sensitive to papain and ß‐mercaptoethanol, but resistant to trypsin, pepsin, lipase, catalase, α‐amylase, organic solvents, detergents, EDTA and heat treatment (90°C/30 min). It was active at a wide pH range (2.0–8.0) and produced at maximum level in de Mann Rogosa Sharpe broth at 32°C with an inoculum amount of 0.1–1.0% and an initial pH of 5.5–7.5. It was active against some Gram‐positive bacteria, including Listeria monocytogenes. It showed bactericidal activity against L. monocytogenes and its molecular weight was approximately 5.8 kDa.     

Evaluation of the psychrotrophic specific signatures for cspA gene and 16S rDNA on the phenotype of Bacillus cereus sensu strictu

This study characterised the psychrotrophic genotypes and phenotypes behaviour of 63 strains of Bacillus cereus sensu stricto isolated from dairy products. The presence of the cspA gene signature and the 16S rDNA mesophilic and/or psychrotrophic specific signatures was evaluated. Among the strains, 25 (39.7%) had the cspA gene signature, 38 (60.3%) had both mesophilic and psychrotrophic 16S rDNA signatures, 24 (38.1%) had only mesophilic and one exhibited only psychrotrophic. No strain grew at 7 °C. The results indicate that the presence of psychrotrophic signatures for cspA gene or the 16S rDNA did not ensure a psychrotrophic behaviour on a B. cereus phenotype.     

Reductionin Listeria monocytogenes,Salmonella enterica and Escherichia coli O157:H7 in vitro and on tomato by sophorolipid and sanitiser as affected by temperature and storage time

Increased consumption of produce by consumers has been attributed to perceived health benefits of postharvest produce. Pathogen control is crucial because periodic occurrences and contamination of tomato and leafy greens have exacerbated food safety risks for consumers. We investigated the effects of temperatures (5 and 25 °C), storage time (30 min and 24 h) for inactivation of Listeria monocytogenes, Salmonella enterica and Escherichia coli O157:H7 by sophorolipid (SL‐p) produced fermentatively using palmitic acid as a co‐substrate at different concentrations in vitro. Reduction in pathogenic bacteria on grape tomato by SL‐p, sanitiser (Lovit) and combinations of SL‐p and sanitiser was determined. Temperature and storage time significantly (P < 0.05) affected pathogen inactivations by SL‐p as pathogen reductions were greater at 25 °C and 24 h than at 5 °C and 30 min of storage. L. monocytogenes was the most sensitive to SL‐p treatment as reductions of 5 log relative to untreated controls were attained at 0.12% of SL‐p. Significant reductions in S. enterica (1.91–3.85 logs) and E. coli O157:H7 (0.87–4.09 logs) were recorded at 2–5% of SL‐p. Lower populations of Salmonella and E. coli O157:H7 were inactivated than L. monocytogenes. On grape tomato, pathogen populations inactivated increased at higher SL‐p levels at 25 °C. Sanitiser and sanitiser + SL‐p reduced bacterial populations on tomato by 5.29–5.76 logs and 0.71–3.3.66 logs, respectively. These results imply the interactions of temperature, storage time and SL‐p significantly (P < 0.05) affected pathogen strain reductions. The combination of SL‐p with sanitiser led to synergistic effect on E. coli O157:H7, but not L. monocytogenes and S. enterica.     

Effects of encapsulation on the viability of probiotic strains exposed to lethal conditions

The effect of microencapsulation on the viability of Lactobacillus casei, L. paracasei, L. acidophilus Ki and Bifidobacterium animalis BB‐12 during exposure to lethal conditions (25% NaCl, pH 3.0 and 55–60 °C) was evaluated. Results demonstrated that survival of probiotic strains to the imposed lethal stress conditions was strain dependent. With the exception of exposure to 25% (w/v) NaCl, L. acidophilus Ki (free and encapsulated cells) demonstrated the highest survival rates through exposure to lethal conditions of temperature and pH. For this probiotic strain exposed to heat, microencapsulated cells expressed a higher heat tolerance at 55 °C than free cells. For the other tested bacteria, in general, encapsulation had no positive effect on survival through the tested lethal conditions.     

Development of vacuum‐dried probiotic milk powder with Bacillus coagulans

The present study explored the possibilities of using Bacillus coagulans as a probiotic culture in vacuum‐dried milk powder. The operational drying temperature at 63 ± 2 °C under 0.7 kg/cm² pressure emerged as the best temperature at which to prepare dried milk powder within 4.5 h with a mean (±SEM) Bacillus coagulans B37 spore count of 8.78 ± 0.03 log cfu/g and moisture content of 4.82 ± 0.12%. The total spore counts in vacuum‐dried milk powder did not change (P > 0.10) at storage temperatures of 7 °C, 37 °C or 45 °C over a three‐month period.     

Effects of High‐Hydrostatic Pressure on Inactivation of Human Norovirus and Physical and Sensory Characteristics of Oysters

The purpose of the study was to determine the effect of high‐hydrostatic pressure (HHP) on inactivation of human norovirus (HuNoV) in oysters and to evaluate organoleptic characteristics of oysters treated at pressure levels required for HuNoV inactivation. Genogroup I.1 (GI.1) or Genogroup II.4 (GII.4) HuNoV was inoculated into oysters and treated at 300 to 600 MPa at 25 and 0 °C for 2 min. After HHP, viral particles were extracted by porcine gastric mucin‐conjugated magnetic beads (PGM‐MBs) and viral RNA was quantified by real‐time RT‐PCR. Lower initial temperature (0 °C) significantly enhanced HHP inactivation of HuNoV compared to ambient temperature (25 °C; P < 0.05). HHP at 350 and 500 MPa at 0 °C could achieve more than 4 log₁₀ reduction of GII.4 and GI.1 HuNoV in oysters, respectively. HHP treatments did not significantly change color or texture of oyster tissue. A 1‐ to 5‐scale hedonic sensory evaluation on appearance, aroma, color, and overall acceptability showed that pressure‐treated oysters received significantly higher quality scores than the untreated control (P < 0.05). Elevated pressure levels at 450 and 500 MPa did not significantly affect scores compared to 300 MPa at 0 °C, indicating increasing pressure level did not affect sensory acceptability of oysters. Oysters treated at 0 °C had slightly lower acceptability than the group treated at room temperature on day 1 (P < 0.05), but after 1 wk storage, no significant difference in sensory attributes and consumer desirability was observed (P > 0.05).     

Synergy between high-pressure, temperature and ascorbic acid on the inactivation of Bacillus cereus

B. cereus strain (CECF 148) was used as a model system in the study of the behaviour of bacillus under high pressure, at temperatures over and below 0 °C and with ascorbic acid added to the culture. Three different assays were carried out in the present experiment. The first assay was performed to observe how B. cereus reacted to pressure shift freezing (PSF) treatment at different subzero temperatures (−8, −12, −20 and −17 °C) and pressures (120, 150, 210 and 350 MPa) in their vegetative form. In the second assay, we observed how different concentrations of ascorbic acid (1, 2, 5 and 20 mM) added to the growing brought decreased B. cereus on its vegetative form. Finally, we tried to inactivate the vegetative and spore form of B. cereus under pressure of 210 MPa at room (20 °C) (HP) and at subzero (−20 °C) (PSF) temperatures, in presence of ascorbic acid (20 mM), added to the growing culture (TSB). The results confirmed that pressures of 210 and 350 MPa at low temperatures (−20 and −17 °C) in the PSF treatment were not enough to inactivate bacillus and only about 10% of B. cereus at the assayed conditions (350 MPa at −17 °C) lost its growth capacity. The presence of ascorbic acid reduced the amount of B. cereus. The initial amount of B. cereus in the vegetative form was 10⁸ to 10⁹ cfu/mL. After HP (210 MPa at 20 °C) and PSF (210 MPa at −20 °C) treatments, the amount of B. cereus decreased by 4 and 2 logarithmic units, respectively. However, in both treatments, the presence of ascorbic acid (20 mM concentration) reduced the B. cereus growth capacity in about 5 logarithmic units. The presence of ascorbic acid in the spore form decreased the amount of B. cereus only by 2 logarithmic units, but without the antioxidant, the values remained close to control. The present research is a contribution to elicit the safety standards of food treated by high pressure.     

Effect of growth temperature on yeast lipid composition and alcoholic fermentation at low temperature

The loss of viability of wine yeast strains due to low-temperature fermentations could be overcome by increasing their stress tolerance and adaptability. Changes in membrane lipid composition are one of the first responses to cold stress. The aim of this study was to analyze the various adaptation mechanisms to low temperatures by comparing the better adapted Saccharomyces species. The viability, vitality, fermentation capacity, and lipid composition of different Saccharomyces species (S. cerevisiae, S. bayanus, S. uvarum, and a hybrid S. cerevisiae/S. uvarum) with different fermentative origins (wine, beer, and baker’s strains together with a laboratory strain) were compared after culturing at low (13 °C) and optimal (25 °C) temperatures. In spite of specific responses of the different strains/species, the results showed that at low temperature, the medium-chain fatty acid and the triacylglyceride content increased, whereas the phosphatidic acid content and the phosphatidylcholine/phosphatidylethanolamine ratio decreased. Only the laboratory strain was not able to ferment the sugars, and after growing at both temperatures, its lipid composition was very different from that of the other strains. The hybrid strain showed the highest sugar consumption at 13 °C and the best vitality whatever the preculture temperature used. The rest of the species needed a preadaptation at low temperature involving a change in their lipid composition to improve their fermentation rate at 13 °C.     

Antimicrobial effects of pressurised carbon dioxide on Brochothrix thermosphacta in broth and foods

Antimicrobial effects of high‐pressure CO₂ on Brochothrix thermosphacta were investigated in a batch system. Inactivation rates increased with increasing pressure, temperature and exposure time. B thermosphacta suspended in physiological saline was completely inactivated under 6.05, 3.02 and 1.51 MPaCO₂ treatment at 35 °C after 10, 25 and 50 min respectively. Two phases were observed in the survival curves. The earlier stage was characterised by a slow rate of decrease in the number of B thermosphacta, which increased sharply at the later stage. B thermosphacta suspended in brain heart infusion broth was completely inactivated under 6.05 MPaCO₂ treatment after 80, 50 and 30 min at 25, 35 and 45 °C respectively. About 6.90 and 5.93 log cycles of B thermosphacta were reduced under 6.05 MPaCO₂ pressure at 45 °C in whole and skimmed milk respectively. The sterilisation effects of 6.05 MPaCO₂ pressure at 45 °C on both B thermosphacta and aerobic plate count were observed after 150 and 120 min respectively in skinned meat. © 2000 Society of Chemical Industry     

Microbiological and physicochemical properties of dry‐cured neck inoculated with probiotic of Bifidobacterium animalis ssp. lactis BB‐12

The effect of inoculum level of Bifidobacterium animalis ssp. lactis BB‐12 probiotic strain and ripening period on the quality of dry‐cured neck was studied. The microbiological parameters (Enterobacteriaceae, LAB and TVC) and physicochemical attributes (pH value, a_w, TBARS index, colour) were determined directly after fermentation at 15 °C for 3 weeks, after 6 and 12 months of ripening at 4 °C. The highest LAB count and a lower pH value were found in the meat inoculated with probiotic strain at 6.6 log cfu g^?1 (B2) followed by inoculation with probiotic strain at 6.3 log cfu g^?1 (B1). Level of inoculation had not had an influence on water activity, TBARS index and total colour parameters. Changes of fat oxidation during half‐year of ripening were limited in probiotic meat samples compared to naturally fermented control meat (C). Based on the results, it can be concluded that the most favourable physicochemical and microbiological parameters of the dry‐cured neck were obtained after 6 months of ripening. At that time, the Bifidobacterium BB‐12 at both levels is a good potential starter for meat fermentation.     

The glycerol and ethanol production kinetics in low-temperature wine fermentation using Saccharomyces cerevisiae yeast strains

Increasing glycerol production in low-temperature wine fermentation is of concern for winemakers to improve the quality of wines. The objective of this study was to investigate the effect of 10 different Saccharomyces cerevisiae on the kinetics of production of glycerol, ethanol and the activities of glycerol-3-phosphate dehydrogenase (GPD) and alcohol dehydrogenase (ADH) in low-temperature fermentation. Ethanol production was influenced by temperature, and it was slightly higher at 13 °C than at 25 °C. Glycerol yields were significantly affected by both temperature and strains. More glycerol was produced at 25 °C than at 13 °C because the activity of GPD was higher at 25 °C than at 13 °C. Glycerol production of the different yeast strains was up to 3.19 and 3.18 g L⁻¹ at 25 and 13 °C, respectively. Therefore, isolating the yeast strains with high glycerol production and adaptation to low-temperature fermentation is still the best method in winemaking.     

Resistance of phages lytic to pathogenic Escherichia coli to sanitisers used by the food industry and in home settings

Phages are potentially useful as antimicrobial agents in food‐processing environments, provided they can remain active upon extended storage and in the presence of sanitisers. Survival of six phages lytic against enteropathogenic (EPEC) and shiga‐toxigenic (STEC) Escherichia coli strains was assessed upon storage at 4 °C, ?20 °C and ?70 °C in phosphate‐buffered‐saline (PBS) and Tris‐magnesium‐gelatine buffer (TMG) for up to 1 year. The phages were also exposed to ethanol, sodium hypochlorite, peracetic acid, quaternary ammonium chloride (biocide A), hydrogen peroxide/peracetic acid/peroctanoic acid (biocide B), p‐toluensulfonchloroamide (biocide D) and alkaline chloride foam (biocide C). Plaque‐forming units remained unchanged when the phages were stored at 4 °C in both buffers tested, but decreased by 3.5 and 5.7 log₁₀ PFU when stored in PBS at ?20 and ?70 °C, respectively. Moreover, TMG seems to be the most protective suspension medium with decreasing temperature because a 1?log₁₀ PFU reduction was observed at ?20 and ?70 °C. Incubation in 100% ethanol for 24 h reduced plaque counts only by 2.5 log₁₀ PFU. In 10 ppm of sodium hypochlorite and in biocide B (0.13%), the counts decreased by ~5 and ~6 log₁₀ PFU after 15 min. Finally, biocide A and D reduced counts by 4 and 2–4 log₁₀ PFU after 30 min and 8 h of incubation, respectively. Phages were completely inactivated only by peracetic acid and biocides C and E. Therefore, the phages evaluated might be potentially applied together with classical sanitisers such as ethanol and industrial biocides A, B and D, in disinfection programs against pathogenic STEC and EPEC strains.     

Characterization of Gram‐Negative Psychrotrophic Bacteria isolated from Italian Bulk Tank Milk

Eighty psychrotrophic bacterial strains, isolated from different northwest Italian bulk tank milks destined for Grana Padano cheese production, were identified by 16S rRNA gene amplification and partial sequence analysis of the rpoB gene. Pseudomonas spp. were the most commonly occurring contaminants, P. fluorescens being the predominant isolated species, along with Enterobacteriaceae, primarily Serratia marcescens. RAPD‐PCR was used to study genetic variability and distinguish closely related strains; a high degree of genetic heterogeneity among the strains was highlighted. All the strains were characterized for their ability to produce proteases, lipases and lecithinases at different temperatures (7, 22, and 30 °C). Forty‐one of the psychrotrophic strains were positive for all the enzymatic activities. The highest number of positive strains for all the incubation temperatures was found for lipolytic activity (59), followed by proteolytic (31) and lecithinase (28) activities, and the enzymatic traits varied among the Pseudomonas and Enterobacteriaceae strains. The proteolytic psychrotrophic strains were screened for the presence of the aprX gene, coding for a heat‐resistant metalloprotease in Pseudomonas spp. The aprX gene was detected in 19 of 63 Pseudomonas strains, and was widespread in the P. fluorescens strains (14/19).     

Antibacterial activity of Ginseng (Panax ginseng C. A. Meyer) stems–leaves extract produced by subcritical water extraction

Antibacterial activities of ginseng extracts produced from ginseng by‐products, stems and leaves, using subcritical water extraction (SWE) at 110, 165 and 190 °C, were evaluated and compared with those of ginseng extracts prepared by hot water and ethanol extraction. The ginseng stems–leaves extract produced by SWE at 190 °C contained the greatest concentration of phenolics (98.4 mg GAE g^?1 of extract). All ginseng extracts inhibited the growth of Bacillus cereus, Salmonella enteritidis, Escherichia coli O157:H7 and Listeria monocytogenes. Among four strains, B. cereus was more sensitive to the ginseng extract by SWE at 190 °C than other bacteria. Cell membranes of bacteria were disrupted by the addition of SWE ginseng extract, observed by transmission electron microscopy (TEM), with the release of cellular contents. These findings provided evidence about the potential utilisation of ginseng stems and leaves by using an environmental friendly extraction process, SWE, to produce ginseng extract for the inhibition of bacteria growth.     

γ‐Glu‐Met synthesised using a bacterial glutaminase as a potential inhibitor of dipeptidyl peptidase IV

This study describes γ‐glutamyl dipeptides as competitive inhibitors of dipeptidyl peptidase‐IV (DPP‐IV), and the feasible synthesis of γ‐Glu‐Met through transpeptidation catalysed by a commercial glutaminase of Bacillus amyloliquefaciens. γ‐Glu‐Met, γ‐Glu‐Leu, γ‐Glu‐Phe, γ‐Glu‐Trp or γ‐Glu‐Tyr exhibited a competitive inhibitory effect on DPP‐IV. The yield of γ‐Glu‐Met was 26.16% under optimised conditions: 200 mm Gln, 20 mm Met, 0.1 U mL^?1 enzyme, pH 9.0, 37 °C and reaction time 3 h. For the first time, the side products containing characteristic sequences, that is poly‐γ‐glutamyl short chains with a terminal Met residue (γ‐Glu‐γ‐Glu‐Met and γ‐Glu‐γ‐Glu‐γ‐Glu‐Met) were identified. The superiority of the commercial glutaminase in the synthesis of DPP‐IV‐inhibitory peptides can enable the application of this novel process for manufacturing γ‐glutamyl‐peptides as potential functional ingredients in the type 2 diabetic diet.     

Potential of aflatoxin B1 production by Aspergillus flavus strains on commercially important food grains

In this study, we investigated the potential of aflatoxin B₁ (AFB1) production by five Aspergillus flavus strains previously isolated from sorghum grains on cereals (barley, maize, rice, wheat and sorghum), oilseeds (peanuts and sesame) and pulses (greengram and horsegram). Five strains of A. flavus were inoculated on all food grains and incubated at 25 °C for 7 days; AFB1 was extracted and estimated by enzyme‐linked immunosorbent assay. All A. flavus strains produced AFB1 on all food grains ranging from 245.4 to 15 645.2 μg kg^?1. Of the five strains tested, strain Af 003 produced the highest amount of AFB1 on all commodities ranging from 2245.2 to 15 645.2 μg kg^?1. Comparatively, the AFB1 accumulation was high on rice grains ranging from 3125.2 to 15 645.2 μg kg^?1, followed by peanuts ranging from 2206.2 to 12 466.5 μg kg^?1. Less AFB1 accumulation was observed in greengram and sesame seeds ranging from 645.8 to 2245.2 and 245.4 to 2890.6 μg kg^?1, respectively. Our results showed that all food grains tested are susceptible to A. flavus growth and subsequent AFB1 production.