Lorenzo Siroli

Lactic acid bacteria and natural antimicrobials to improve the safety and shelf-life of minimally processed sliced apples and lamb's lettuce

Outbreaks of food-borne disease associated with the consumption of fresh and minimally processed fruits and vegetables have increased dramatically over the last few years. Traditional chemical sanitizers are unable to completely eradicate or kill the microorganisms on fresh produce. These conditions have stimulated research to alternative methods for increasing food safety. The use of protective cultures, particularly lactic acid bacteria (LAB), has been proposed for minimally processed products. However, the application of bioprotective cultures has been limited at the industrial level. From this perspective, the main aims of this study were to select LAB from minimally processed fruits and vegetables to be used as biocontrol agents and then to evaluate the effects of the selected strains, alone or in combination with natural antimicrobials (2-(E)-hexenal/hexanal, 2-(E)-hexenal/citral for apples and thyme for lambs lettuce), on the shelf-life and safety characteristics of minimally processed apples and lambs lettuce. The results indicated that applying the Lactobacillus plantarum strains CIT3 and V7B3 to apples and lettuce, respectively, increased both the safety and shelf-life. Moreover, combining the selected strains with natural antimicrobials produced a further increase in the shelf-life of these products without detrimental effects on the organoleptic qualities.

Effects of sub-lethal concentrations of thyme and oregano essential oils, carvacrol, thymol, citral and trans-2-hexenal on membrane fatty acid composition and volatile molecule profile of Listeria monocytogenes, Escherichia coli and Salmonella enteritidis.

The aim of this work was to investigate the modifications of cell membrane fatty acid composition and volatile molecule profiles of Listeria monocytogenes, Salmonella enteritidis, Escherichia coli, during growth in the presence of different sub-lethal concentrations of thyme and oregano essential oils as well as carvacrol, thymol, trans-2-hexenal and citral. The results evidenced that the tested molecules induced noticeable modifications of membrane fatty acid profiles and volatile compounds produced during the growth. Although specific differences in relation to the species considered were identified, the tested compounds induced a marked increase of some membrane associated fatty acids, particularly unsaturated fatty acids, trans-isomers, and specific released free fatty acids. These findings can contribute to the comprehension of the stress response mechanisms used by different pathogenic microorganisms often involved in food-borne diseases in relation to the exposure to sub-lethal concentrations of natural antimicrobials.

Evaluation of the effect of carvacrol on the Escherichia coli 555 metabolome by using 1H-NMR spectroscopy.

Cultures of Escherichia coli 555 were grown at four levels of carvacrol (0-2 mM) and the E. coli endo-metabolome was extracted and measured by (1)H NMR spectroscopy. The results show that glucose concentration is going up with concentration of carvacrol and so do formate until the highest concentration is reached, from which point it suddenly decreases. This is interpreted as if the bacteria are increasingly unable to further metabolize glucose and as if the bacteria increasingly shifts with higher levels of carvacrol toward sugar fermentation as carbon source, until the level of carvacrol reaches a level (2.00 mM), where the E. coli must give up. Additionally, the multivariate Principal Component Analysis suggests that the adaptation occurring at sub-lethal doses of carvacrol is different from that occurring at higher doses.

Combined effects of high pressure homogenization treatment and citral on microbiological quality of apricot juice

High pressure homogenization (HPH) technique is able to significantly reduce spoilage microbiota in fruit juice. On the other hand, aroma compounds and essential oils can have a key role in the microbial stability of these products. For this reason, the aim of this work was to evaluate the combined effects of an aroma compound (citral, used at a concentration of 50 mg/l) and HPH treatments (performed at 100 MPa for 1-8 successive passes) on the inactivation dynamics of Saccharomyces cerevisiae SPA strain inoculated in apricot juices at level of about 4.5 log CFU/ml. Moreover, growth of surviving yeast cells was measured during the storage of the treated juice at 10°C and pH, water activity, viscosity and volatile molecule profile of apricot juice were studied. Since citral had been diluted in ethanol before the addition to juice, also samples with only ethanol added at the same volume used to dissolve citral were considered. The results showed that yeast cell viability decreased with the increases of passes at 100 MPa and the relationship between yeast cell loads and number of passes at 100 MPa followed a linear trend. In addition, the effect of HPH treatment can be notably potentiated throughout the presence of citral and ethanol, increasing the time necessary to reach a spoilage threshold during storage. The volatile profiles of the juices added with citral showed a substitution by yeast metabolism of this aldehyde with molecule characterized by a lower antimicrobial activity such as alcohols. The HPH treatments had also a significant effect on pH and viscosity of apricot juices while did not affect a(w).

Use of homogenisation pressure to improve quality and functionality of probiotic fermented milks containing Lactobacillus rhamnosus BFE 5264

The effects of 60-MPa milk homogenisation treatment were investigated on the viability of the probiotic strain Lactobacillus rhamnosus BFE5264, when used as yoghurt co-starter, as well as on the aroma profile, texture and microstructure of the resulting set-type fermented milks. The results demonstrated that the combined use of homogenisation pressure and co-inoculation of the probiotic strain with the traditional yoghurt starters allowed the reduction in the product coagulation time, the increase of the probiotic strain viability and the improvement of the product volatile molecule profiles. The rheological indices and the microstructure results indicated the positive effects of the milk homogenisation treatment on the product viscosity index and exopolysaccharide production.

Determination of Antibacterial and Technological Properties of Vaginal Lactobacilli for Their Potential Application in Dairy Products

Functional foods could differently affect human health in relation to the gender. Recent studies have highlighted the anti-Candida and anti-Chlamydia activities of some Lactobacillus strains isolated from the vagina of healthy women. Considering these important beneficial activities on womens health, the preparation of functional food containing active vaginal lactobacilli can represent a great scientific challenge for the female gender. In this context, the aim of this work was to study some functional and technological properties of 17 vaginal strains belonging to the species Lactobacillus crispatus, Lactobacillus gasseri, and Lactobacillus vaginalis in the perspective to include them in dairy products. The antagonistic activities against the pathogenic and spoilage species associated to food products and against the principal etiological agents of the genitourinary tract infections were evaluated. Moreover, the vaginal lactobacilli were characterized for their antibiotic resistance, and for their fermentation kinetics and viability during the refrigerated storage in milk. Finally, the volatile molecule profiles of the obtained fermented milks were determined. The results showed that several strains, mainly belonging to the species Lactobacillus crispatus, exhibited a significant antagonistic activity against spoilage and pathogenic microorganisms of food interest, as well as against urogenital pathogens. All the vaginal lactobacilli showed antimicrobial activity against strains belonging to the foodborne pathogenic species Listeria monocytogenes, Listeria innocua, Eenterococcus faecalis and Escherichia coli. In addition, most of the Lactobacillus strains were active toward the main pathogens responsible of vaginal and urinary tract infections including Staphylococcus aureus, Enterococcus faecium, Gardnerella vaginalis, and Proteus mirabilis. The antimicrobial activity can be attributed to the high production of organic acids. The fermentation kinetics in milk indicated the unsuitability of these lactobacilli as fermentation starters for the industrial production of dairy products. However, some strains, belonging to the species Lactobacillus crispatus and Lactobacillus gasseri, maintained a high viability in pasteurized milk at 4°C for over a month, showing their potential application as adjunct cultures for the production of female gender foods. These data represent a first step for the set-up of a new functional dairy product, directed to the women well-being, contributing also to innovate the dairy sector.

Microencapsulation of functional strains by high pressure homogenization for a potential use in fermented milk

This study was aimed to evaluate the potential of high pressure homogenization for the microencapsulation of two probiotic lactic acid bacteria, Lactobacillus paracasei A13 and Lactobacillus salivarius subsp. salivarius CET 4063 to produce functional fermented milks. Microcapsules of the considered functional microorganisms were obtained by HPH treatments at 50MPa in the presence of sodium alginate and vegetable oil. The microencapsulated microorganisms were then inoculated as adjuncts to produce fermented milks. As controls were used fermented milks in which the two probiotic lactobacilli were inoculated without encapsulation. The viability of the strains was monitored during almost 2months of refrigerated storage. The survival of lactic acid bacteria after the gastric-duodenal simulated test was determined. Fermented milk texture parameters, the presence of exo-polysaccharides and the production of volatile molecules were also evaluated over storage. The microcapsules, for both the considered probiotic strains, were homogeneous and with a size<100μM and therefore did not adversely affect the sensory properties of the fermented milks. The encapsulation decreased the hyperacidity phenomena generally related to the inclusion of probiotic microorganisms in fermented milks. The lower acidity of the products due to the microencapsulation was fundamental for the improvement of the viability of the starter culture and the sensory characteristics of the products. The microencapsulation conditions increased the resistance to the simulated digestion processes, although the strain Lb. paracasei A13 generally showed a higher resistance to the gastric barrier respect to Lb. salivarius CECT 4063. By contrast, the data obtained showed a reduction of EPS production by the microencapsulation. The volatile profiles showed specific profiles in relation to the probiotic strain used and microencapsulation process. In conclusion, the results of this study underlined the applicative potential of HPH microencapsulation of probiotic microorganisms to produce fermented milk with improved functionality and with enhanced sensory properties.

Effects of chitosan based coatings enriched with procyanidin by-product on quality of fresh blueberries during storage

The aim of this work was to evaluate the efficacy of an innovative edible coating, based on chitosan from mushrooms enriched with procyanidins extracted from grape seeds, on fresh blueberry quality maintenance, (weight loss, pH, dry matter, colour, firmness and antioxidant activity) and microbial growth, during 14 days of storage at 4 °C. For weight loss, pH and dry matter no relevant differences were detected among the control and the differently coated samples at each considered storage time. Chitosan and chitosan + procyanidins coatings promoted a slight decrease of luminosity and an increase of blue hue colour of blueberry samples during the whole storage period. The use of coating promoted an increase in the antiradical activity that was the highest in blueberries coated with chitosan + procyanidins. Microbiological analysis results indicated that the chitosan-based coated samples had a significantly higher yeast and mould growth inhibition compared to the uncoated sample.

Optimization of Vacuum Impregnation with Calcium Lactate of Minimally Processed Melon and Shelf-Life Study in Real Storage Conditions

Vacuum impregnation (VI) is a processing operation that permits the impregnation of fruit and vegetable porous tissues with a fast and more homogeneous penetration of active compounds compared to the classical diffusion processes. The objective of this research was to investigate the impact on VI treatment with the addition of calcium lactate on qualitative parameters of minimally processed melon during storage. For this aim, this work was divided in 2 parts. Initially, the optimization of process parameters was carried out in order to choose the optimal VI conditions for improving texture characteristics of minimally processed melon that were then used to impregnate melons for a shelf-life study in real storage conditions. On the basis of a 23 factorial design, the effect of Calcium lactate (CaLac) concentration between 0% and 5% and of minimum pressure (P) between 20 and 60 MPa were evaluated on color and texture. Processing parameters corresponding to 5% CaLac concentration and 60 MPa of minimum pressure were chosen for the storage study, during which the modifications of main qualitative parameters were evaluated. Despite of the high variability of the raw material, results showed that VI allowed a better maintenance of texture during storage. Nevertheless, other quality traits were negatively affected by the application of vacuum. Impregnated products showed a darker and more translucent appearance on the account of the alteration of the structural properties. Moreover microbial shelf-life was reduced to 4 d compared to the 7 obtained for control and dipped samples.

Potential of Natural Antimicrobials for the Production of Minimally Processed Fresh-Cut Apples

Background: Minimally processed fruit are susceptible to microbial spoilage and loss of sensory quality. In this experimental work, in order to increase the shelf-life and to maintain quality parameter (texture and colour) of sliced apples (Malus communis, var. Golden delicious), the use of natural antimicrobials was proposed. Materials and methods: Natural antimicrobials were purposed to produce fresh-cut apples. Hexanal, citral, and combinations of hexanal+citral, citron essential oil+carvacrol, citral+2-(E)-hexenal, citral+citron essential oil and hexanal+2- (E)-hexenal were used in dipping step. After treatment, sample were stored at 6°C in ordinary atmosphere. During storage, yeast and lactic acid bacteria were monitored. Also volatile and electronic nose profiles, colour and texture analyses were considered. The samples were compared with a control (apples dipped in ascorbic and citric acid solution). Results: Yeast cell loads showed that natural antimicrobials changed naturally occurring yeast growth parameters. The combination of citron+carvacrol prolonged the yeast lag phase of 6 d comparing with control, while citral and hexanal+2-(E)- hexenal decreased the maximum reached yeast cell load and growth rate, respectively. After 8 d, samples with hexanal+2-(E)- hexenal and citral showed equivalent or even better quality attributes compared to the controls, suggesting that this approach is a useful tool for fresh-cut apple production. Highlight: Natural antimicrobial can be useful for the dipping of minimally processed apples Citral and hexanal+2-(E)-hexenal prolonged the apples shelf-life. Apples treated with Citral and hexanal+2-(E)-hexenal had good quality attributes.