Proteolytic pattern and organic acid profiles of probiotic Cheddar cheese as influenced by probiotic strains of Lactobacillus acidophilus,Lb. paracasei,Lb. casei or Bifidobacterium sp.

Cheddar cheeses were produced with starter lactococci and Bifidobacterium longum 1941, B. lactis LAFTI^® B94, Lactobacillus casei 279, Lb. paracasei LAFTI^® L26, Lb. acidophilus 4962 or Lb. acidophilus LAFTI^® L10 to study the survival of the probiotic bacteria and the influence of these organisms on proteolytic patterns and production of organic acid during ripening period of 6 months at 4 °C. All probiotic adjuncts survived the manufacturing process of Cheddar cheese at high levels without alteration to the cheese-making process. After 6 months of ripening, cheeses maintained the level of probiotic organisms at >8.0 log₁₀ cfu g⁻¹ with minimal effect on moisture, fat, protein and salt content. Acetic acid concentration was higher in cheeses with B. longum 1941, B. lactis LAFTI^® B94, Lb. casei 279 and Lb. paracasei LAFTI^® L26. Each probiotic organism influenced the proteolytic pattern of Cheddar cheese in different ways. Lb. casei 279 and Lb. paracasei LAFTI^® L26 showed higher hydrolysis of casein. Higher concentrations of free amino acids (FAAs) were found in all probiotic cheeses. Although Bifidobacterium sp. was found to be weakly proteolytic, cheeses with the addition of those strains had highest concentration of FAAs. These data thus suggested that Lb. acidophilus 4962, Lb. casei 279, B. longum 1941, Lb. acidophilus LAFTI^® L10, Lb. paracasei LAFTI^® L26 and B. lactis LAFTI^® B94 can be applied successfully in Cheddar cheese.     

Effect of acidification on the activity of probiotics in yoghurt during cold storage

The viability of Lactobacillus acidophilus LAFTI^® L10, Bifidobacterium lactis LAFTI^® B94, and L. paracasei LAFTI^® L26 and their proteolytic activities were assessed in yoghurt at different termination pH of 4.45, 4.50, 4.55, and 4.60 in the presence of L. delbrueckii ssp. bulgaricus Lb1466 and Streptococcus thermophilus St1342 during 28 days of storage at 4 °C. All strains achieved the recommended level of 6.00 log cfu g⁻¹ of the product with L. acidophilus LAFTI^® L10 and L. paracasei LAFTI^® L26 exceeding the number to 8.00 and 7.00 log cfu g⁻¹, respectively. Lactobacilli strains showed a good cellular stability maintaining constant concentration throughout storage period regardless of termination pH. On the other hand, the cell counts of B. lactis LAFTI^® B94 decreased by one log cycle at the end of storage. The presence of probiotic organisms enhanced proteolysis significantly in comparison with the control batch containing L. delbrueckii ssp. bulgaricus Lb1466 and S. thermophilus St1342 only. The proteolytic activity varied due to termination pH, but also appeared to be strain related. The increased proteolysis improved survival of L. delbrueckii ssp. bulgaricus Lb1466 during storage resulting in lowering of pH and production of higher levels of organic acids, which might have caused the low cell counts for B. lactis LAFTI^® B94.     

Effect of cryoprotectants,prebiotics and microencapsulation on survival of probiotic organisms in yoghurt and freeze-dried yoghurt

The survival of probiotic microorganisms including Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus rhamnosus and Bifidobacterium spp. was evaluated in yoghurt and freeze-dried yoghurt after processing and storage. The effectiveness of microencapsulating probiotic organisms as well as adding cryoprotectants and prebiotics in improving their viability was also investigated. The viability of Bifidobacterium infantis 17930 and L. rhamnosus GG was reduced by 0.07 log, while that of L. casei 1520 and Bifidobacterium longum 1941 was reduced by 0.28 and 0.39 log, respectively. There was a 7% improvement in the viability of L. casei 1520 when cryoprotectant ‘Unipectine™ RS 150’ was added at 2.5% (w/v). The prebiotic ‘Raftilose^®P95’ when added at 1.5% w/v to yoghurt improved the viability of the combined selected probiotic organisms by 1.42 log during four weeks of storage at 4 °C. Microencapsulation with alginate improved viability of combined selected probiotic organisms by 0.31 log in freeze-dried yoghurt stored at 21 °C.     

Probiotic stability of yoghurts containing Jerusalem artichoke inulins during refrigerated storage

Experimentally prepared Jerusalem artichoke inulins (JAI) were compared with two commercial chicory root inulins for their prebiotic potentials in media broth model and growth-sustaining ability in non-fat yoghurts. Experimental yoghurts were made with 12% reconstituted skim milk (RSM) supplemented with 4% inulin powders, inoculated with mixed cultures of Lactobacillus casei LC-01, Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus (1:0.5:0.5 based on supplier’s recommendation) and incubated overnight at 37 °C. Non-supplemented yoghurt was prepared from 16% RSM and used as control. The survival and acidifying activity of lactic and probiotic cultures in all yoghurts were investigated on weekly intervals during the shelf life of 28 days at 4 °C. Incorporation of JAI resulted in improved viability of LC-01, maintaining >7.0 log CFU/g during cold storage but did not affect the viability of yoghurt bacteria in comparison with the control.     

Rheology,spontaneous whey separation,microstructure and sensorial characteristics of probiotic yoghurts enriched with passion fruit fiber

Beyond demonstrated beneficial health attributes, passion fruit rinds are a by-product of the fruit pulp industry, rich in total dietary fiber, particularly pectin. The aim of this study was to evaluate the influence of the addition of passion fruit fiber on the whey formation, rheological parameters, microstructure and sensorial characteristics of probiotic yoghurts. Skim milk bases enriched with 1% of passion fruit fiber or not were heat treated and inoculated with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus, and divided into four groups according to the probiotic strain added — Lactobacillus acidophilus strains L10 and NCFM and Bifidobacterium animalis subsp. lactis strains Bl04 and B94. Fermentations were performed until the pH reached 4.5. Rheological characteristics of yoghurts were determined by a rotational rheometer in two cycles of shear rate ranging from 0 to 15 s^? 1 in both upward and downward curves. Sensorial analysis of passion fruit fiber yoghurts, either without any probiotic or co-fermented by L. acidophilus L10 or B. animalis subsp. lactis Bl04, was evaluated against a control yoghurt without fiber. Photomicrographs of freeze-dried yoghurts were made by field-emission scanning electron microscope (SEM). Thixotropy of enriched yoghurts was higher than that of their respective controls in the two cycles of shear rate. Apparent viscosity was significantly higher in fiber yoghurts co-fermented by the lactobacilli than in their controls at the end of cold storage. Photomicrographs demonstrated that in passion fruit fiber yoghurts the casein gel was more compact and overlaid the fiber, while filaments of exopolysaccharides were more frequent in control yoghurts. Appearance, odor and color of the passion fruit fiber yoghurts received scores as ‘good’, and the intensity of the passion fruit flavor was considered weak by the sensory assessors. Results indicate that the passion fruit fiber is an almost neutral ingredient for the design of new high value-added yoghurt.     

Chemical analysis and sensory evaluation of Cheddar cheese produced with Lactobacillus acidophilus,Lb. casei,Lb. paracasei or Bifidobacterium sp.

The sensory properties of probiotic Cheddar cheeses made using Lactobacillus acidophilus 4962, Lb. casei 279, Bifidobacterium longum 1941, Lb. acidophilus LAFTI^® L10, Lb. paracasei LAFTI^® L26 or B. lactis LAFTI^® B94 were assessed after ripening for 9 months at 4 °C. Probiotic cheeses except those with Lb. acidophilus 4962 were significantly different (P<0.05) from the control without any probiotic organism. Acceptability of probiotic cheese with Lb. casei 279 was significantly lower (P<0.05) than that of the control cheese with bitterness and sour-acid taste as the major defects. Concentration of acetic acid in probiotic cheeses was higher (P<0.05) than the control cheese. Vinegary scores did not influence the acceptability of the cheeses (P>0.05). Increased proteolysis in probiotic cheeses did not influence the Cheddary attribute scores (P>0.05). There were positive correlations (P<0.05) between the scores of bitterness and the level of water-soluble nitrogen.     

Rheological,microstructural and sensory characterization of low-fat and whole milk set yoghurt as influenced by inulin addition

The effect of inulin addition (0–4%) upon texture and microstructure of set yoghurt with different levels of fat (0.2%–3.5%) was investigated. A two-factor experimental design with four treatments was used for data analysis. Skimmed milk with various inulin and cream concentrations was standardized to 4% protein content, homogenized, heated to 92 °C and fermented at 42 °C until a pH of 4.6 was reached. The chemical composition, pH, consistency and microstructure properties of the yoghurts were analysed after 6 days of storage at 5 °C. The statistical analysis showed that inulin and fat significantly affected the rheological and sensory results. Higher yield stress, “firmness” and “creaminess” values were observed in yoghurt produced with higher inulin additions, whereas the pH value was not affected. A significant correlation was found between yield stress and sensory determined firmness (r = 0.91). The microstructure examined by confocal laser scanning microscopy (CLSM) was only slightly affected by the concentrations of inulin in the range studied, possibly due to weak protein interactions between the inulin and the milk protein network.     

Production of traditional Greek yoghurt using Lactobacillus strains with probiotic potential as starter adjuncts

Lactobacillus plantarum ACA-DC 146 and L. paracasei subsp. tolerans ACA-DC 4037 were examined for their potential application as adjuncts in the production of traditional Greek set-type yoghurt. Both strains displayed low milk acidification activity, while no inhibition was observed towards or from the yoghurt starters used. Yoghurt produced with L. paracasei subsp. tolerans ACA-DC 4037 exhibited the best sensory properties, with a rich traditional smooth taste, and the strain was selected for further trials. Yoghurt produced with this strain as an adjunct had good physicochemical properties. After 2 weeks of refrigerated storage, microbial loads (>7.0 log cfu g⁻¹) were in accordance with international recommendations and guidelines for probiotic and starter cultures in milk products. Increasing the microbial load further, using concentrated and encapsulated inocula (10–11 log cfu g⁻¹), gave yoghurt with long fermentation times and poor organoleptic properties.     

Incorporation of selected nutraceuticals and probiotic bacteria into a fermented milk

Yoghurts were produced from a base milk containing three important nutraceuticals, namely ω-3-fatty acids, isoflavones and phytosterols. The cultures employed to make the yoghurts were single probiotic strains of Lactobacillus gasseri or Bifidobacterium infantis and, to achieve a short production time, a two-stage fermentation procedure was used with Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus providing the rapid acidification. Yoghurts containing counts of >1.0×10⁸ cfu mL⁻¹ of the individual probiotics and high counts of the traditional species from yoghurt were awarded overall scores for sensory acceptability >4.0 out of 5.0; the nutraceuticals appeared to have no adverse effect on flavour. Storage trials at 5 °C showed that the viability of the probiotic cultures was retained over 15 days.     

Characteristics of stirred low-fat yoghurt as affected by high pressure

The effects of high pressure on the physicochemical, chemical, microbiological and sensory characteristics of stirred low-fat yoghurt were studied. Laboratory-made yoghurts were treated at high pressure (100–400 MPa) for 15 min at 20°C. No significant changes in pH and total organic acids were observed after pressuring the yoghurt. Pressures over 200 MPa prevented post-acidification of the yoghurt during chilled storage. Pressurized yoghurts exhibited higher viscosity and amino acid contents than did the untreated controls, and the differences were maintained after chilled storage. High-pressure treatments at 300 and 400 MPa reduced the number of viable cells of lactobacilli to below the legal minimum permitted in many countries. Significant differences in sensory characteristics between untreated and pressurized yoghurts (200–300 MPa) were detected after chilled storage.     

Modifying the microstructure of low-fat yoghurt by microfluidisation of milk at different pressures to enhance rheological and sensory properties

The effects of microfluidisation of milk at different pressures, prior to heat treatment, on structural and sensory properties of low-fat stirred yoghurt, were investigated. Low-fat yoghurts prepared from microfluidised milk were compared with low-fat (1.5%) and full-fat (3.5%) control yoghurts made with homogenised (20/5 MPa) milk. The microstructure of low-fat yoghurts prepared with microfluidised milk consisted of smaller and more uniform fat globules, well incorporated into more interconnected fat-protein gel networks, compared with those of control yoghurts. This modification in microstructure caused significant changes in gel particle size, sensory profile and rheological behaviour. Microfluidisation increased the gel particle size, gel strength and viscosity; marked beneficial effects were found at higher pressures (50–150 MPa). Microfluidising milk at 50–150 MPa increased the gel strength by 171–195% and viscosity by 98–103%, creating low-fat yoghurts with creaminess and desirable texture properties similar to, or better than, full-fat conventional yoghurt.     

ACE-inhibitory activity of probiotic yoghurt

In this study, the in vitro angiotensin-converting enzyme (ACE)-inhibitory (ACE-I) activity of peptide fractions from different yoghurt batches was assessed. Inhibition of ACE activity resulted in an overall antihypertensive effect. Yoghurts were prepared either using a sole yoghurt culture including Lactobacillus delbrueckii ssp. bulgaricus Lb1466 and Streptococcus thermophilus St1342, or L. acidophilus L10, L. casei L26 and Bifidobacterium lactis B94 in addition to yoghurt culture. ACE-I activity was determined at weekly intervals during 28 days of cold storage. Peptide fractions showing high ACE-I activity were further purified using multiple-steps of RP-HPLC. All probiotic yoghurts showed appreciable ACE-I activity during initial stages of storage compared with the control yoghurt, with a significant (p<0.05) decrease afterwards. The ACE-I activity ranged from IC₅₀ of 103.30–27.79 μg mL⁻¹ with the greatest ACE inhibition achieved during first and third week of storage. The in vitro ACE-I activity could be related to the peptide liberation via degradation of caseins. In total, 8 ACE-I peptides were characterized originating from α_s2-casein (1), κ-casein (2) and β-casein, of which two well-known ACE-inhibiting peptides, namely Val–Pro–Pro (VPP) and Ile–Pro–Pro (IPP), were identified. These peptides are already used in commercial products.     

Sensory and rheological screening of exopolysaccharide producing strains of bacterial yoghurt cultures

Yoghurts were produced with 24 different cultures differing in their ability to produce exopolysaccharides (EPS). Rheological and sensory analyses showed large differences in texture properties in the yoghurt samples. EPS production was found to have a major effect on the texture properties, but varying textures within the EPS±groups were also found. Yoghurts fermented with EPS-producing cultures showed increased mouth thickness and ropiness and tended to be creamier than yoghurts without these cultures; in contrast, these yoghurts had the lowest syneresis and highest gel firmness (initial gel strength before the yoghurt was subjected to shear). Correlations between rheological parameters and sensory texture attributes included G* correlating with gel firmness, while viscosity measured at 241 s⁻¹ correlated with mouth thickness. An interesting viscometry relationship was found between ropiness and hysteresis loop area.     

Physical properties and microstructure of yoghurts supplemented with milk protein hydrolysates

The physical properties and the microstructure of yoghurts containing probiotic bacteria, and supplemented with milk protein hydrolysates, were studied. Three casein hydrolysates and three whey protein hydrolysates were added to milk at a concentration ranging from 0.25 to 4 g L⁻¹. The milks were then fermented with either of two different cultures. The resulting yoghurts with added hydrolysates were compared to the control yoghurt without supplementation. For both cultures, addition of hydrolysates decreased the complex viscosity and graininess in yoghurts. The addition of hydrolysates also reduced fermentation time. Microstructural observations showed a more open and less branched structure in yoghurts when milk protein hydrolysates were incorporated. The difference in fermentation time between milks with different levels of added hydrolysates could partially explain the differences in microstructure and physical properties of the final yoghurts.     

Effect of long‐chain inulin on the texture profile and survival of Lactobacillus paracasei ssp. paracasei in set yoghurts during refrigerated storage

The effects of adding inulin at 20 g/L as a fat replacer and probiotic bacteria on the physicochemical and textural characteristics of yoghurt were studied. The ability of long‐chain inulin to improve the probiotic (Lactobacillus paracasei ssp. paracasei) bacteria viability in yoghurt was investigated. The addition of inulin made the texture (firmness, cohesiveness, adhesiveness and gumminess) of skimmed yoghurt similar to that of whole yoghurt, demonstrating the role of inulin as a fat replacer. However inulin increased syneresis and did not influence the viability of probiotic bacteria in the yoghurts. The addition of probiotic bacteria in yoghurts improved syneresis and increased firmness and gumminess.     

Effects of somatic cell counts in milk on physical and chemical characteristics of yoghurt

The quality of plain stirred yoghurt produced from whole milk with somatic cell counts (SCC) at low (147,000 cells mL⁻¹), intermediate (434,000 cells mL⁻¹) and high (1,943,000 cells mL⁻¹) levels was examined. Each milk treatment was obtained from selected cows, according to its SCC status and milk composition. Yoghurt samples were analysed on days 1, 10, 20 and 30 after production. Analyses included pH, acidity, fat, lipolysis (expressed as free fatty acids, FFA), proteolysis and apparent viscosity. Viscosity of high SCC yoghurt was higher (P<0.05) than the low SCC yoghurt on days 10, 20 and 30 of storage. High SCC yoghurt also had higher FFA content (P<0.05). SCC did not affect pH, acidity, fat content and proteolysis of the yoghurt (P>0.05). Results indicate that SCC in milk increases the lipolysis in the resulting yoghurt during storage for 30 d.     

Viability of Lactobacillus acidophilus in rice bran‐enriched stirred yoghurt and the physicochemical and sensory characteristics of product during refrigerated storage

This study was aimed at investigating the fortification of probiotic yoghurt with rice bran to increase nutritional properties of the product. The different levels of rice bran (0.3%, 0.6%, 0.9% and 1.2%) were incorporated into milk. The yoghurt samples were produced after pasteurisation, addition of starter culture and 1% Lactobacillus acidophilus suspension (6 × 10⁸ CFU mL^?1) and incubation. During sample storage in refrigerator, the viability of L. acidophilus, viscosity and physicochemical and sensory properties of product were investigated. Rice bran significantly increased the viability of L. acidophilus (P < 0.05). In addition, all probiotic yoghurts incorporating rice bran indicated higher viscosity and acidity and lower pH and syneresis compared to plain yoghurts. Furthermore, increments in rice bran incorporation levels resulted in a reduction in consumers' sample preferences. In general, the addition of rice bran at a suitable level could increase L. acidophilus viability and improve quality attributes of yoghurt.     

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.     

Rheological properties of stirred yoghurt as affected by gel pH on stirring,storage temperature and pH changes after stirring

The rheological properties of stirred yoghurt were studied as a function of the delay between milk heat-treatment and inoculation (0, 1 and 2 days), of pH in the acid gel on stirring (4.4, 4.7, and 5.0), of the storage temperature (4, 12, and 20 °C) for 24 h following stirring and of over-acidification (allowed or inhibited). At low pH values, the gels exhibited higher elastic modulus (G′) and fracture strength. They yielded stirred yoghurts with higher G′ and viscosity, and higher increase in G′ and viscosity during storage (“rebodying”). Rebodying was only partially explained by over-acidification and cooling. Changing the storage temperature had no impact on the evolution of G′ after stirring; hydrophobic interactions were therefore probably not involved in rebodying. Electrostatic interactions seemed to play a major role in rebodying, as pH on stirring was the significant factor.