Productions and monomer compositions of exopolysaccharides by Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains isolated from traditional home-made yoghurts and raw milk

In this study, a total of forty‐five strains of lactobacilli and streptococci were determined exopolysaccharide (EPS) production in skim milk and Man Rogosa and Sharpe (MRS)/M17 medium, viscosity and proteolytic activity. The exopolysaccharide production by lactobacilli strains during growth in MRS medium was twenty‐one to 211 mg L^?1, while in skim milk was to thirty‐six to 315 mg L^?1. The EPS production by streptococci strains during growth in M17 medium was sixteen to 114 mg L^?1, while in skim milk was to twenty‐four to 140 mg L^?1. The EPS production of strains was lower in MRS/M17 medium than skim milk. Results showed that it was not clear correlation between the viscosity and EPS production of some strains. All strains were shown proteolytic activity. Positive correlations between exopolysaccharide production and proteolytic activity in skim milk were found some strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. These results indicated that the high exocellular protease‐producing strains can produce high EPS in skim milk. The monomer compositions of the EPSs formed by selected five strains were analysed. Mannose dominated (99–100%) on the EPS produced by L. delbrueckii subsp. bulgaricus and S. thermophilusstrains (except L. delbrueckii subsp. bulgaricus 22) in skim milk and MRS/M17 medium. Besides, the EPSs of strains in skim milk contained small amount of lactose.     

Study of the effects of spray‐drying on the functionality of probiotic lactobacilli

Three probiotic lactobacilli strains were spray‐dried in 20% (w/v) skim milk and submitted to a simulated gastrointestinal digestion. Fresh or spray‐dried cultures were administered to mice for 5 and 10 days, and Immunoglobulin A (IgA)‐producing cells were enumerated in the small intestine by immunohistochemistry. Spray‐drying significantly enhanced the resistance of Lactobacillus paracasei A13 and Lactobacillus casei Nad to a simulated gastrointestinal digestion (0.96 and 1.95 log orders, respectively), compared with fresh cultures. Also, a significant higher number of IgA‐producing cells were induced by spray‐dried cultures compared with fresh cultures. Spray‐drying is a suitable, but strain‐dependent, technological process for the development of probiotic cultures in skim milk with increased functionality.     

Fermented beverage emulsion based on donkey milk with sunflower oil

Sunflower oil was added to donkey milk to increase the low energetic intake and improve both texture and healthy characteristics. This milk‐based emulsion was fermented using a binary association of two strains of Streptococcus thermophilus showing distinctive abilities. One strain (St 907) produced ropy exopolysaccharide (EPS), whereas the other strain (St 563) produced folic acid. Fermentation was stopped at pH 5.0 to prevent whey separation. The refrigerated product was stable at least for 20 days and showed improved viscoelastic properties due to the EPS combined with the oil droplets. The volatile profile was constituted of ketones, aldehydes, alcohols and fatty acids, mainly originated from microbial fermentation, whereas few other compounds suggested the presence of oxidative processes. The folic acid content in the fermented beverage was increased by more than 10 times (2.03 ± 0.17 μg/100 mL) with respect to that found in donkey milk (0.16 ± 0.03 μg/100 mL).     

Optimisation of bambara groundnut water extract and skim milk composition as cryoprotectant for increasing cell viability of Lactobacillus spp. using response surface methodology

Four legume water extracts, that is bambara groundnut, soya bean, red kidney bean and black bean as well as skim milk, were examined for their effectiveness in protecting Lactobacillus rhamnosus GG and Lactobacillus fermentum SK5 during the freeze‐drying and storage. Bambara groundnut water extract (BGWE) showed promising cryoprotective activity that was comparable to skim milk. BGWE and skim milk at 2–10% w/v and 5–20% w/v individually produced survival rates for both strains of 87–88%. To further optimise the synergistic cryoprotective medium, response surface methodology was employed. The optimal combination was 4.93% w/v BGWE and 11.68% w/v skim milk for L. rhamnosus GG and 5.17% w/v BGWE and 11.36% w/v skim milk for L. fermentum SK5 with survival rates of 95.17% and 94.36%, respectively. The storage life of freeze‐dried cells of both probiotics at 4 °C and 30 °C for 6 months was markedly improved when they were produced with these optimal combinations.     

Detection of Salmonella spp., Yersinia enterocolitica,Listeria monocytogenes and Campylobacter spp. by real‐time multiplex PCR using amplicon DNA melting analysis and probe‐based assay

Syto9 and probe‐based multiplex real‐time PCR assays for simultaneous detection of a group of foodborne pathogens (named SYLC group), targeting Salmonella spp. (invA gene), Yersinia enterocolitica (ystA gene), Listeria monocytogenes (hly gene) and Campylobacter spp. (rrna gene), have been developed. The Syto9 assay generates amplicon DNA melting curve with four peaks of 86.5 ± 0.5, 84 ± 0.5, 81.5 ± 0.5 and 90.5 ± 0.5 °C corresponding Salmonella spp., Y. enterocolitica, L. monocytogenes and Campylobacter spp. targets, respectively. The sensitivities of the Syto9 and TaqMan assays in artificially inoculated chicken wing rinses were in a range of 3.2 × 10² to 3.1 × 10⁴ and 9.8 × 10² to 1.9 × 10⁴ colony‐forming units per millilitre, respectively, depending on the pathogen. All tested target strains (n = 100) were correctly detected by the both assays, whereas nontarget strains (n = 100) demonstrated no cross‐reactivity representing 100% specificity. The assays are suitable for application in qualitative and quantitative detection of SYLC group pathogens in food matrices.     

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.     

Effect of fermentation temperature on the properties of exopolysaccharides and the acid gelation behavior for milk fermented by Streptococcus thermophilus strains DGCC7785 and St-143

We investigated the effect of fermenting milk with 2 strains (DGCC7785 and St-143) of Streptococcus thermophilus, which are known to produce different types of exopolysaccharide (EPS) structures. The yields and physical properties of these ropy EPS were monitored during the fermentation of milk at different temperatures. We wanted to understand how these types of EPS properties affected yogurt gelation. Reconstituted skim milk was fermented at 33, 39, or 45°C until pH values reached 5.2, 4.9, 4.7, and 4.5. Molar mass of ropy EPS samples was determined using size exclusion chromatography coupled with multiangle laser light scattering. Rheological properties of fermented milk gels were analyzed using small-strain dynamic oscillatory measurements. In both strains, concentrations of ropy EPS increased during fermentation and at all temperatures. Fermentation times, by both strains, were shortest at 45°C and longest at 33°C. For both strains, molar mass of ropy EPS ranged from 2 to 4 × 10⁶ g/mol during fermentation. A major proteinaceous contaminant that was co-isolated with the ropy EPS fraction by our isolation method was identified as a milk-derived phosphoglycoprotein PP3. Increase in fermentation temperature from 33 to 45°C significantly decreased the storage modulus values (from 170 to 41 Pa) for milk gelled by strain DGCC7785, whereas the gels made with St-143 had very low storage modulus values (11–17 Pa) regardless of fermentation temperatures. For both strains, the values of maximum loss tangent in the milk gels increased with fermentation temperature; the maximum loss tangent occurred at higher pH values when milk was fermented by strain DGCC7785. The specific type of EPS produced appeared to be responsible for the differences in yogurt texture rather than the concentration or molar mass of the EPS.     

Influence of exopolysaccharide‐producing cultures on the volatile profile and sensory quality of low‐fat Tulum cheese during ripening

The objective of this investigation was to compare the composition and changes in the concentration of volatiles in low‐fat and full‐fat Tulum cheeses during ripening. Tulum cheese was manufactured from low‐ or full‐fat milk using exopolysaccharide (EPS)‐producing or non‐EPS‐producing starter cultures. A total of 82 volatile compounds were identified belonging to the following chemical groups: acids (seven), esters (21), ketones (14), aldehydes (six), alcohols (14) and miscellaneous compounds (20). The relative amounts of acids, alcohols and aldehydes increased in the cheeses made with EPS‐producing cultures during 90 days of ripening. Differences were found in the volatile profile of full‐fat Tulum cheese compared with the low‐fat variant, especially after 90 days of ripening. Exopolysaccharide‐producing cultures changed the volatile profile, and the EPS‐producing cultures including Streptococcus thermophilus + Lactobacillus delbrueckii subsp. bulgaricus + Lactobacillus helveticus (LF‐EPS2) produced cheese with higher levels of methyl ketones and aldehydes than the non‐EPS cultures. In the sensory analysis, full‐fat Tulum cheeses and the cheese produced with the EPS‐producing culture containing Lb. helveticus (LF‐EPS2) were preferred by the expert panel. It was concluded that the use of EPS‐producing starter cultures in the manufacture of low‐fat Tulum cheese had the potential to improve the flavour.     

Impact of β-glucan on debittering,bioaccessibility and storage stability of skim milk fortified with shrimp oil nanoliposomes

Shrimp oil was encapsulated in nanoliposomes and fortified into skim milk. Shrimp oil nanoliposomes (SONL) were thermodynamically stable when added into skim milk at 10 mL 100 mL⁻¹. Mild bitterness in fortified skim milk caused by the SONL was masked by adding β-glucan at various levels (0.05–0.2 g 100 mL⁻¹). With the addition of SONL, fortified skim milk appeared more reddish in colour due to the presence of astaxanthin. Addition of β-glucan resulted in the increase in viscosity of the fortified milk by forming network of junction zones. During the storage of skim milk fortified with SONL and 0.1 g 100 mL⁻¹ β-glucan at 4 °C for 15 days, no major quality changes took place. Simulated in vitro digestion studies revealed that 45.41 g 100 g⁻¹ eicosapentaenoic acid (EPA) and 48.86 g 100 g⁻¹ docosahexaenoic acid (DHA) from shrimp oil were bioaccessible for absorption in the gut after digestion.     

Determination of the relationship between oxalate degradation and exopolysaccharide production by different Lactobacillus probiotic strains

The aim of this study was to evaluate the protective effects of exopolysaccharide (EPS) production on resistance to gastrointestinal digestive conditions and oxalate by probiotic strains of the species Lactobacillus rhamnosus, Lactobacillus fermentum and Lactobacillus brevis. The oxalate‐degrading ability of the strains was determined using an enzymatic assay. The correlation between oxalate degradation rate and EPS production was not significant (P > 0.05); however, the high‐EPS‐producing L. fermentumIP5 and L. brevisYG7 strains showed high oxalate‐degrading activity, whereas the low‐EPS‐producing strain L. fermentumBP5 demonstrated low oxalate‐degrading activity. The present findings suggest that dietary supplementation with the probiotic strain L. fermentumIP5 could be a promising strategy for the prevention of oxalate stone disease.     

Effect of the main ingredients of Rhizoma gastrodiae on mycelial biomass and exopolysaccharide productions by submerged culture of Grifola frondosa

Various concentrations of the main ingredients of Rhizoma gastrodiae including gastrodin (GA) and p‐hydroxybenzyl alcohol (HA) as well as p‐hydroxylbenzaldehyde (HBA) were separately added into the cultural medium of Grifola frondosa in submerged culture. The results showed that the additive concentration of gastrodin from 0.10 to 0.35 g L^?1, p‐hydroxybenzyl alcohol from 0.05 to 0.30 g L^?1 and p‐hydroxylbenzaldehyde from 0.05 to 0.30 g L^?1 can significantly promote the EPS production by submerged culture of G. frondosa (P < 0.05). Among them, the addition of p‐hydroxylbenzaldehyde at 0.15 g L^?1 exhibited the best results, and the EPS productions reached 2.2511 ± 0.0378 g L^?1, with an increase of 45.03% compared with the control (without additives). Furthermore, all of the additives had an effective stimulatory effect on mycelial biomass. In addition, there is an important and novel breakthrough that p‐hydroxylbenzaldehyde was more effective than R. gastrodiae extract on increasing the EPS productions.     

Streptococcus thermophilus strains of plant origin as dairy starters: Isolation and characterisation

Streptococcus thermophilus strains have been isolated mainly from dairy environments. To prospect for new strains of S. thermophilus, isolation was made from different plant sources. In this study, 74 plant isolates were characterised as S. thermophilus by a polyphasic approach and by 16S rRNA sequencing. The isolates were further evaluated for their physiological and biochemical properties. Plant isolates exhibited good acid production and varying proteolytic activity. All the isolates showed acetaldehyde and capsular exopolysaccharide (EPS) production and few revealed diacetyl production. In contrast to industrial strains, six isolates were able to ferment galactose and 25 were found to have no urease activity. Both the plant isolates and reference dairy cultures were found to possess similar physiological and biochemical properties and can be considered for developing new starters.     

Research on some factors influencing acid and exopolysaccharide produced by dairy propionibacterium strains isolated from traditional homemade Turkish cheeses

In this study, a total of 32 isolated strains and 5 reference strains of dairy propionibacteria were analyzed for acid and exopolysaccharide (EPS) production in skim milk and yeast extract-lactate broth (YEL) media in order to investigate the physiological background and preservative role of acid and EPS. The effects of final culture pH and optical density on acid and EPS production were also determined. On average, all strains produced more acid and reached lower final pH values in skim milk than in YEL medium. While the correlations obtained between the acid produced by propionibacterium strains and their final culture pH in skim milk medium were significant (P < 0.01), no correlations were found between optical density, final pH, and produced acid in YEL medium. Sixteen isolated and five reference strains of propionibacteria were tested further for the ability to produce propionic and acetic acids. On average, Propionibacterium freudenreichii subsp. shermanii and P. freudenreichii subsp. freudenreichii strains produced higher amounts of propionic and acetic acids than did Propionibacterium jensenii in YEL medium. The acid produced by these strains may be used as a preservative in the food industry for replacement or reduction of the increasing use of chemical additives. The EPS production by propionibacterium strains during growth in YEL medium was 72 to 168 mg/liter, while in skim milk it was 94 to 359 mg/liter. The monomer compositions of the EPSs formed by the six selected dairy propionibacteria strains were analyzed. The EPSs may have applications as food grade additives and viscosity-stabilizing agents.     

Molecular Characterization and Identification of Bioactive Peptides Producing Lactobacillus sps. Based on 16S rRNA Gene Sequencing

In the present study, 3 bacterial cultures were isolated from faecal samples of human infant. The biochemical traits showed similarity with Lactobacillus sps and 16S rRNA sequence analyses, confirmed as Lactobacillus plantarum, Lactobacillus casei, and Lactobacillus rhamnosus. The cultures were screened for their proteolytic activity and good ability to release peptides from milk proteins was found. Hence, these bacteria were used as a proteolytic starter culture for the fermentation of skim milk and whey for the liberation of small peptides. Bioactive nature of the peptides released from whey and skim milk was tested, and results demonstrated that peptides obtained after fermentation of whey and skim milk by Lactobacillus strains showed antimicrobial activity against all the pathogens causing food borne infections in humans. These peptides also indicated antioxidant as well as ACE (angiotensin-converting enzymes) inhibitory activity.     

Effect of exopolysaccharides and proteolytic activity of Lactococcus lactis subsp. cremoris strains on the viscosity and structure of fermented milks

Two exopolysaccharide (EPS)-producing Lactococcus lactis subsp. cremoris strains (B35 and B891) were used to study the effect of the kinetics of EPS production and bacterial proteolytic activity on the structure of milk gels and the viscosity of stirred milk gels. Strains were grown at 20 °C in milk containing either yeast extract or casitone and at 30 °C in either milk alone or milk containing casitone. Lactococcal counts, pH decrease and production and molecular characteristics (molar mass and radius of gyration) of both EPSs were followed during milk fermentation. The level of proteolysis in the fermented milks was determined after 24 h of incubation. The results obtained showed that the yield of EPS and the timing of EPS production during milk-gel formation were the most important factors that influenced the structure of the milk gels and the viscosity of the stirred product. The proteolytic activity of the strains did not seem to play any significant role.     

Initial study on high pressure jet processing using a modified waterjet on physicochemical and rennet coagulation properties of pasteurized skim milk

Developments in material science and engineering have enabled high pressure jet (HPJ) processing at >500 MPa. The objective of this study was to characterize the physicochemical properties of pasteurized skim milk processed at pressures from 0 to 500 MPa using waterjet technology, to explore novel uses of this technology for milk. The apparent particle size of casein micelles increased from approximately 180–220 nm in milk processed from 0 to 200 MPa to approximately 280 nm in milk treated at 500 MPa. All milk samples were shelf-stable up to 14 days at 4 °C. The viscosity of skim milk processed at 400 and 500 MPa almost doubled (4.2 mPa s) when compared with control unprocessed milk (2.2 mPa s). Increasing HPJ processing pressure changed the structure–function properties of the casein micelles and no rennet-induced coagulation was observed for milk processed at 500 MPa.     

Influence of heat impact in reconstituted skim milk on the properties of yoghurt fermented by ropy or non‐ropy starter cultures

The paper describes studies on the influence of heat impact in reconstituted skim milk on chemical and functional properties of yoghurt products. Reconstituted skim milk was heated for 20 min at 85°C, 90°C, or 95°C. Ropy (producing exopolysaccharides, EPS) or non‐ropy strains of S. thermophilus and L. delbrueckii subsp. bulgaricus were used as starter culture for yoghurt manufacture. The studies have shown that the fermentation times decreased with increasing heat impact when the ropy starter culture was used, while they remained to a far extent unchanged if the non‐ropy starter culture was applied. The lactic acid contents of the yoghurt products were in the same range when the milk was heated at 85°C or 90°C, while they were different when milk was heated at 95°C. There was a tendency visible that an increase in preheating leads to increased L (+)‐ and decreased D (–)‐lactic acid contents if the non‐ropy culture was applied. Using the ropy culture, it was vice versa. A slightly decrease in proteolysis with increasing heat impact was to be noted with both starter cultures. Concerning the relation of proteolysis to acidification, the fermentation process could be subdivided into three sections with different slopes if the non‐ropy starter culture was used, while a linear relation was found if the ropy starter culture was applied. Regarding final product characteristics it was found that the functional properties of yoghurt decreased with increasing heat impact when the ropy starter culture was applied, while they remained to a far extent unchanged when the non‐ropy starter culture was used. It can be concluded from these studies that a preheating of milk at a temperature of 85°C (20 min) is optimal in regard to final yoghurt product characteristics.     

Properties of low-fat stirred yoghurts made from high-pressure-processed skim milk

Physical properties of stirred yoghurt made from reconstituted skim milk that was high-pressure (HP)-treated at 100, 250 or 400 MPa, at 25, 70 or 90 °C, for 10 min, prior to inoculation with yoghurt cultures, were studied; portions of milk HP-treated at 25 °C were also heat-treated at 90 °C for 10 min before or after pressure treatment. Control yoghurts were made from skim milk given a heat treatment at 90 °C for 10 min. Fermentation time was not affected by treatment applied to the milk. HP treatment of skim milk at 25 °C, before or after heat treatment, gave stirred yoghurts of similar viscosities to that made from conventionally heat-treated milk. Lower viscosities were obtained when stirred yoghurts were made with milk HP-treated at elevated temperatures. A model is proposed to correlate properties of yoghurt with HP/heat-induced changes in interactions and structures of protein in the milk samples.Industrial relevanceTo meet end user expectations, the dairy industry needs to diversify its product range by tailoring specific functionalities. To meet these expectations, new processing methods such as high-pressure processing are of interest for their potential to achieve specific and/or novel functionalities and/or improve efficiencies, including reduced chemical and water use. In this paper, an investigation of the use simultaneous pressurization and heating of milk before the manufacture of stirred yoghurt is presented.