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A comprehensive analysis of safety and functional features of six Weissella and Periweissella type strains was performed. Our data demonstrated the probiotic potential of these species, indicating the P. beninensis type strain as the best candidate based on its potential probiotic features and the safety assessment.

The application of mathematical modeling to study and characterize lactic acid bacterial strains with pro-technological and functional features has gained attention in recent years to solve the problems relevant to the variabilities of the fermentation processes of sourdough. Since the key factors contributing to the sourdough quality are relevant to the starter strain growth and its metabolic activity, in this study, the cardinal growth parameters for

A metabolic feature of lactic acid bacteria (LAB) is the production of exopolysaccharides (EPSs), which have technological and functional properties of interest to the food sector. The present study focused on the characterization of the

Bacterial strains belonging to Lacticaseibacillus paracasei species are generally used as starters in food fermentations and/or as probiotics. In the current study, the growth cardinal parameters of four L. paracasei strains (IMPC2.1, IMPC4.1, P40 and P101), isolated from table olives or human source, were determined. Strains were grown in liquid medium and incubated at several temperatures (10 values from 5.5°C–40°C) and pH (15 values from 3.2 to 9.1) along the growth range. The cardinal temperature model was used to describe temperature effects on the maximum specific growth rate of L. paracasei whereas new equations were developed for the effect of pH. The estimated Tmin values ranged between −0.97°C and 1.95°C and were lower than 0°C for strains IMPC4.1 and P101. Strain P40 was able to grow in the most restricted range of temperature (from 1.95°C to 37.46°C), while strain IMPC4.1 was estimated to survive at extreme conditions showing the lowest pHmin. Maximum specific growth rates of L. paracasei IMPC2.1 in white cabbage (Brassica oleracea var. capitata) were used to calculate the correction factor (Cf) defined as the bias between the bacterial maximum specific growth rate in broth and in the food matrix. A simple bi-linear model was also developed for the effect of temperature on the maximum population density reached in white cabbage. This information was further used to simulate the growth of L. paracasei strains in cabbage and predict the time to reach the targeted probiotic level (7 log10 CFU/g) using in silico simulations. This study demonstrates the potential of the predictive microbiology to predict the growth of beneficial and pro-technological strains in foods in order to optimize the fermentative process.

A metabolic feature of lactic acid bacteria (LAB) is the production of exopolysaccharides (EPSs), which have technological and functional properties of interest to the food sector. The present study focused on the characterization of the Weissella cibaria strain C43-11, a high EPS producer in the presence of sucrose, in comparison with a low-producing strain (C2-32), and on possible genetic regulatory elements responsible for the modulation of dextransucrase (dsr) genes expression. NMR analysis of the polymeric material produced by the C43-11 strain indicated the presence of dextran consisting mainly of a linear scaffold formed by α-(1–6) glycosidic linkages and a smaller amounts of branches derived from α-(1–2), α-(1–3), and α-(1–4) linkages. Molecular analysis of the dsr genes and the putative transcriptional promoters of the two strains showed differences in their regulatory regions. Such variations may have a role in the modulation of dsr expression levels in the presence of sucrose. The strong upregulation of the dsr gene in the C43-11 strain resulted in a high accumulation of EPS. This is the first report showing differences in the regulatory elements of the dsr gene in W. cibaria and indicates a new perspective of investigation to identify the regulatory mechanism of EPS production.

The application of mathematical modeling to study and characterize lactic acid bacterial strains with pro-technological and functional features has gained attention in recent years to solve the problems relevant to the variabilities of the fermentation processes of sourdough. Since the key factors contributing to the sourdough quality are relevant to the starter strain growth and its metabolic activity, in this study, the cardinal growth parameters for pH, temperature (T), water activity (aw), and undissociated lactic acid of the sourdough strain Lactiplantibacillus plantarum ITM21B, were determined. The strain growth, pH, organic acids (lactic, acetic, phenyllactic, and hydroxy-phenyllactic), total free amino acids, and proteins were monitored during fermentation of a liquid sourdough based on wheat flour and gluten (Bio21B) after changing the starting T, pH, and inoculum load. Results demonstrated that the different fermentation conditions affected the strain growth and metabolite pattern. The organic acid production and growth performance were modeled in Bio21B, and the resulting predictive model allowed us to simulate in silico the strain performances in liquid sourdough under different scenarios. This mathematical predictive approach can be useful to optimize the fermentation conditions needed to obtain the suitable nutritional and technological characteristics of the L. plantarum ITM21B liquid sourdough.

Exopolysaccharides (EPS) are known for their positive contribute to the technological properties of many foods, including bakery products. These molecules can be obtained performing piloted fermentation with lactic acid bacteria (LAB). In order to select strains able to produce EPS, a screening test in agar medium containing sucrose, fructose or glucose as carbohydrate source was performed on 21 LAB strains. Results allowed to select 8 Weissella cibaria, 2 Weissella confusa and 2 Leuconostoc spp. strains as EPS producers only in the presence of sucrose. A further screening in liquid medium enriched with sucrose (10%) (mMRS_S) indicated the W. cibaria strain C43-11 as the higher EPS producer. The selected strain was used to develop liquid sourdoughs (LSs) with dough yield (DY) 500, fermented for 15 h and based on wheat flour and wheat gluten or pseudocereals (quinoa or amaranth) in ratio 1:1, in the presence or not of sucrose at 3% (w/w, LS weight), in comparison to Lactobacillus plantarum ITM21B, a strain not producing EPS in mMRS_S. Results indicated that the use of pseudocereals favored the EPS production. Formulations were optimized by modifying DY (500 or 250), sucrose concentration (3 or 6%) and flour ratio. LSs were characterized for the content of organic acids (lactic, acetic, phenyllactic, OH-phenyllactic), pH, TTA, EPS, viscosity, total protein degradation and protein pattern. The highest EPS production (20.79 g/Kg) and viscosity (1168 mPa s) were obtained in LS (DY 250, sucrose 6%) based on quinoa flour and started with C43-11 strain. The LS was characterized by the presence of phenyllactic and OH-phenyllactic acids, protein degradation by 51.7% and proteins in the range 14-80 kDa. In these conditions, also strain ITM21B was able to produce EPS at level of 4.61 g/Kg and to degrade proteins by 53.8% in LS based on wheat and quinoa flours (1:1) (DY250 and sucrose 3%). Therefore, results demonstrated that the use of selected conditions (flour type, DY, sucrose) can stimulate specific attributes of strains making them suitable for production of short fermented (15h) liquid sourdoughs which can be used as bread improvers.

Production of the traditional Italian Focaccia bread (FcBread) requires reformulation to meet the new rules for healthier foods. We tested the applicability of a short-fermented (15 h) liquid sourdough (LS) (dough yield, DY, 250), obtained using quinoa (Q) or amaranth (Am) flour fermented with a Weissella cibaria strain (C43-11) producing exopolysaccharides (EPS) as fat replacer in yeast-leavened FcBread. The LSs were applied during the FcBread-making process at industrial pilot plant, reducing the added fat amount by 20% (FcBread-QLS and FcBread-AmLS) and products were compared with reference samples not containing EPS. All the products were analysed for physico-chemical properties (pH, TTA, organic acids, protein content and profile, instrumental colour), textural properties, sensory quality and glycemic index. The application of both pseudo-cereal-based W. cibaria LSs in FcBread allowed obtaining products with a reduced amount of fat, increased protein amount, mainly due to the albumin and glutenin/glutelin fractions, reduced glycemic index, improved texture and preserved in the traditional sensory profile of 'focaccia'.

A pectin coated dehydrated apple snack containing ≥9 log colony forming units/20 g portion of the probiotic Lactobacillus paracasei IMPC2.1, was developed. The strain was incorporated into the apple slices using vacuum impregnation (IP1) or soaking/stirring (IP2); apple slices were then coated with citrus pectin and dehydrated at 4 °C for 14 days. Microbial application led to colonization of the apple surface, as seen using scanning electron microscope images, and all probiotic products showed comparable radical scavenging activity and total phenol content. IP2 led to better bacterial survival during 30 days storage at 4 °C and color parameters comparable to the fresh apple. The simulated gastro-intestinal (GI) digestion suggested the ability of the strain to survive the digestive process. This inclusion/coating/dehydration process led to a probiotic snack, which could successfully deliver bacterial cells in a viable form to the GI tract.

The doctoral thesis is focused on the application of lactic acid bacteria (LAB, CNR-ISPA Collection), selected for their peculiar biotechnological features (antimicrobial, probiotic, proteolytic, EPS production), to food preparation in order to improve their quality. The activities performed so far have concerned: A) use of LAB selected strains in food process to extend their shelf-life (processed fish), to provide probiotic properties (probiotic dehydrated apple snack), to improve nutritional/functional properties of bakery products (reduced addition of salt or fat); B) study of growth behaviour of selected LAB strains to optimize fermentation processes through predictive microbiology

The aim of the current study was to identify quality indicators of fat (14.50 ± 0.75%) and low-fat (4.79 ± 0.63%) raw ground beef by monitoring changes in physicochemical and microbiological parameters during aerobic refrigerated storage, such as water-holding capacity, pH, thiols, carbonyl compounds, thiobarbituric acid reactive substances (TBARS), metmyoglobin, deoxymyoglobin, oxymyoglobin color indices, pseudomonads, Brochothrix thermosphacta, and total viable counts. Meat packaged in air-permeable polyethylene plastic film was stored under controlled isothermal conditions (0, 5, 10, and 15 °C). A population level of pseudomonads equal to 7.0 ± 0.5 log10 colony forming units (CFU)/g was considered as the potential spoilage level. Using principal component analysis, samples were distinguished on the basis of their microbial load. A significant positive correlation between microbial population and carbonyls, metmyoglobin, TBARS, water-holding capacity, and a negative correlation with thiols and color parameters (L* , chroma) were observed. Two different approaches were followed to estimate the quality status of samples: (i) the partial least square (PLS) regression with R2 of 0.93 and root mean square error prediction of 0.44 for pseudomonads, using the above physicochemical characteristics as the dominant input variables, which allowed prediction of the microbiological status of ground beef regardless of time-temperature storage profile and fat content, and (ii) a square-root-type model (adjusted R2 of 0.952) that satisfactorily predicted the growth of spoilage pseudomonads under isothermal and dynamic conditions, regardless of the above physicochemical changes. The above results suggest that depending on the available input data, the two modeling approaches can accurately (and complementarily) assess quality of aerobically stored ground beef. PRACTICAL APPLICATION: Changes in appearance and quality of fat and low-fat raw ground beef are associated with physicochemical alteration and/or microbial growth. The study provides two different modeling approaches that can be integrated in an intelligent interface of the refrigerator having specific colorimetric and/or temperature sensors, to evaluate in a convenience way the quality of stored meat thus reducing domestic waste: the partial least square model was based on physicochemical parameters (particularly chroma, metmyoglobin, and thiobarbituric acid reactive substances), while the square root model was based on the time-temperature conditions during storage.

The use of protective microbial strains in combination with modified atmosphere packaging (MAP) and refrigerated storage on the shelf life of tuna burgers was investigated. Preliminary, the protective ability of three lactic acid bacteria (LAB) cultures (Lactobacillus casei, Lactobacillus paracasei, and Lactobacillus plantarum) have been assessed on ready-to-cook tuna burgers. Among them, L. paracasei showed the best preserving performance and significantly controlled both aerobic mesophilic bacteria and Pseudomonas spp. growth. Subsequently, the efficacy of the selected LAB culture under MAP conditions (5% O2 and 95% CO2) was assessed evaluating microbial and sensory quality, as well as volatile aldehyde content. Results indicated that the shelf life of burgers containing L. paracasei and packaged under MAP was 4 days longer than the control (shelf life about 6 days) and that the applied procedure represents an effective approach for the mild preservation of fish products.

The study aimed at improving the nutritional profile of yeast leavened salt reduced sliced bread and puccia type bread fortified with a wheat-based Lactobacillus plantarum ITM21B fermentation product (Bio21B). The protein content of bread made under laboratory conditions was increased by using: (i) chickpea flour (CF) (15% wt/wt flour) and Bio21B or (ii) the Bio21B containing a fungal protease to favour the gluten hydrolysis. Products showed increased protein and total amino acid content and improved protein digestibility. Moreover, the formula significantly affected the protein pattern of breads which, according to the results of the microfluidic two-dimensional electrophoresis (μ2DE) protein pattern, were discriminated as observed by the PCA plot. The use of CF was validated at industrial pilot plant producing salt reduced sliced bread and puccia type bread. The resulting products showed improved nutritional profile and a sensory quality comparable to the company’s products containing salt.