Probiotic cheese production using Lactobacillus casei cells immobilized on fruit pieces

Lactobacillus casei cells were immobilized on fruit (apple and pear) pieces and the immobilized biocatalysts were used separately as adjuncts in probiotic cheese making. In parallel, cheese with free L. casei cells and cheese only from renneted milk were prepared. The produced cheeses were ripened at 4 to 6°C and the effect of salting and ripening time on lactose, lactic acid, ethanol concentration, pH, and lactic acid bacteria viable counts were investigated. Fat, protein, and moisture contents were in the range of usual levels of commercial cheeses. Reactivation in whey of L. casei cells immobilized on fruit pieces after 7 mo of ripening showed a higher rate of pH decrease and lower final pH value compared with reactivation of samples withdrawn from the remaining mass of the cheese without fruit pieces, from cheese with free L. casei, and rennet cheese. Preliminary sensory evaluation revealed the fruity taste of the cheeses containing immobilized L. casei cells on fruit pieces. Commercial Feta cheese was characterized by a more sour taste, whereas no significant differences concerning cheese flavor were reported by the panel between cheese containing free L. casei and rennet cheese. Salted cheeses scored similar values to commercial Feta cheese, whereas unsalted cheese scores were significantly lower, but still acceptable to the sensory panelists.     

Shelf life of Crescenza cheese as measured by electronic nose

The shelf life of Crescenza, a traditional Italian soft cheese, was measured using classical analysis and a commercial electronic nose. Two lots of samples directly supplied by a manufacturer at the beginning of their commercial life were stored at 2 constant temperatures (8 and 15°C) and analyzed until their respective expiration dates. Among the physicochemical parameters, pH, acidity, hue, and apparent yield rheological index appeared to be the best predictors of the quality decay. Changes in these indices were described with a sigmoidal transition function allowing definition of a loose and a severe shelf-life protocol, based on the trend of first and second time derivatives. A time range of 1 to 3 d at 15°C and 4 to 8 d at 8°C was accordingly assessed to maintain the freshness of Crescenza cheese. The quality decay of cheese aroma was evaluated by inspecting the headspace fingerprint of the same set of samples using the electronic nose. Sample classification through the aroma fingerprint confirmed the predicted shelf-life time ranges. A clear discrimination between “fresh,” “aged,” and “very aged” samples was obtained using principal components analysis, cluster analysis, and linear discriminant analysis statistical techniques. The predictive ability of the linear discriminant analysis classification model was confirmed by considering a new set of cheese samples purchased at the beginning of their commercial life from a local market and analyzed until their expiration date.     

Rennet paste from lambs fed a milk substitute supplemented with Lactobacillus acidophilus: effects on lipolysis in ovine cheese

The present work was undertaken to evaluate the effects of Lactobacillus acidophilus supplementation of a milk substitute on the features of lamb rennet paste used for cheese making. Lipolysis in cheese manufactured with rennet paste from lambs receiving supplemented milk was also evaluated. Lambs were subjected to 3 different feeding regimens (mother suckling, MS; artificial rearing, AR; and artificial rearing with 7 log₁₀ cfu/mL of Lb. acidophilus supplementation of the milk substitute, ARLb) and slaughtered at 20 and 40 d of age for each feeding treatment. Abomasa of the lambs were processed to rennet paste. Microbial loads, enzymatic activities (chymosin, pepsin, and lipases), and renneting characteristics of the lamb rennet paste were determined. Free fatty acids and conjugated linoleic acids were detected in cheese at 60 d of ripening. Addition of 7 log₁₀ cfu/mL of Lb. acidophilus to the milk substitute was carried out successfully. Total recovery of viable cells was recorded in milk supplied daily to the lambs in the ARLb group. The ARLb rennet had greater amounts of lactobacilli than did the MS or AR rennet, irrespective of the slaughter age of the lambs, and the ARLb rennet had higher concentrations of lactococci when lambs were slaughtered at 40 d of age. Chymosin and lipase activities were also higher in ARLb rennet than in MS or AR rennet from lambs slaughtered at an older age. Milk supplementation of ARLb lambs resulted in improved coagulating ability of the rennet and enhanced cheese lipolysis after 60 d of ripening. A reduction of all free fatty acids was observed in all cheeses when passing from 20 to 40 d of slaughter of the lambs. Conjugated linoleic acids were more abundant in ARLb cheeses at both 20 and 40 d. Therefore, supplementation of the milk substitute with Lb. acidophilus improved the enzymatic features of rennet and the healthful and nutritional characteristics of it the ovine cheese. Moreover, the addition of lactobacilli to the milk substitute made it possible to increase the slaughter age of lambs without detrimental effects on rennet characteristics.     

添加双歧杆菌的干酪

双歧杆菌是对人体有益的微生物,干酪的环境有利于双歧杆菌的长期存活;着重介绍了将双歧杆菌加入到干酪方面的研究进展以及干酪中双歧杆菌的精确计数方法。     

Sensory evaluation of probiotic Minas fresh cheese with Lactobacillus acidophilus added solely or in co-culture with a thermophilic starter culture

Sensory acceptance of formulations of probiotic Minas fresh cheese was investigated. Cheeses were prepared and supplemented with Lactobacillus acidophilus (T1 – probiotic), Lactobacillus acidophilus + Streptococcus thermophilus (T2 – probiotic + starter) or produced with no addition of cultures (T3 – control). Sensory acceptance tests were performed after 7 and 14 days of storage at 5 °C, using a 9‐point hedonic scale (1 = dislike extremely; 9 = like extremely). After 7 days, no significant difference was detected among cheeses T1, T2 and T3 (P > 0.05). After 14 days, cheeses T1 and T2 presented higher acceptance and differed significantly from cheeses T3. Cheeses T3 presented significant difference between 7 and 14 days of storage (P < 0.05), whereas probiotic cheeses T1 and T2 were stable in the same period (P > 0.05). The addition of L. acidophilus, either solely or in co‐culture with a thermophilic starter culture, resulted in good acceptance of Minas fresh cheese, improving sensory performance of the product during storage.     

Effect of the inoculation level of Lactobacillus acidophilus in probiotic cheese on the physicochemical features and sensory performance compared with commercial cheeses

The complex metabolism of probiotic bacteria requires several technological options to guarantee the functionally of probiotic dairy foods during the shelf life. This research aimed to evaluate the effect of the supplementation of increasing amounts of Lactobacillus acidophilus (0, 0.4, or 0.8 g/L of milk) on the physicochemical parameters and sensory acceptance of Minas fresh cheese. In addition, the sensory acceptance of probiotic cheeses was assessed using a consumer test and compared with commercial cheeses (conventional and probiotic). High counts (9.11 to 9.42 log cfu/g) of L. acidophilus were observed throughout the shelf life, which contributed to the maintenance of its probiotic status and resulted in lower pH values and greater production of organic acids. The probiotic cheeses presented lower scores for appearance, aroma, and texture compared with conventional cheeses. Internal preference mapping explained almost 60% of the total variation of the data and showed a large number of consumers concentrated near the conventional cheeses, demonstrating greater preference for these samples. The findings indicated that some negative sensory effects could occur when high level of supplementation with L. acidophilus is used in probiotic cheese processing.     

高压处理的霉菌干酪的蛋白水解

对自制的白霉干酪采用高压处理工艺,分析其蛋白水解的情况,并与未经高压处理干酪进行比较。通过测定2种干酪表面和内部的pH4.6-SN(%TN)、12%TCA-SN(%TN)和FAA(mg/g干酪)评价蛋白水解程度,结果表明:高压处理使pH4.6-SN、12%TCA-SN增加,对FAA无影响;高压处理使pH值升高,影响部分微生物酶的活力;高压处理能降低疏水作用,使酶更容易作用于暴露的敏感键。     

Microbiological,immunological, and histological changes in the gut of Salmonella Enteritidis-challenged rats fed goat cheese containing Lactobacillus rhamnosus EM1107

Cheeses are able to serve as suitable matrices for supplying probiotics to consumers, enabling appropriate conditions for bacteria to survive gastric transit and reach the gut, where they are assumed to promote beneficial processes. The present study aimed to evaluate the microbiological, immunological, and histological changes in the gut of Salmonella Enteritidis-challenged rats fed goat cheese supplemented with the probiotic strain Lactobacillus rhamnosus EM1107. Thirty male albino Wistar rats were randomly distributed into 5 experimental groups with 6 animals each: negative (NC) and positive (PtC) control groups, control goat cheese (CCh), goat cheese added with L. rhamnosus EM1107 (LrCh), and L. rhamnosus EM1107 only (EM1107). All animals, except NC group were challenged with Salmonella Enteritidis (10⁹ cfu in 1 mL of saline through oral gavage). Microbial composition was assessed with high-throughput 16S rRNA sequencing by means of Illumina MiSeq (Illumina, San Diego, CA). Nuclear factor kappa B (NF-κB) from the animal cecum tissue was determined by real-time PCR and interleukins (TNF-α, IL-1β, IL-10, and IFN-γ) by means of ELISA. Myeloperoxidase and malondialdehyde levels were determined biochemically. The administration of the L. rhamnosus EM1107 probiotic strain, either as a pure culture or added to a cheese matrix, was able to reduce Salmonella colonization in the intestinal lumen and lessen tissue damage compared with rats from PtC group. In addition, the use of cheese for the probiotic strain delivery (LrCh) was associated with a marked shift in the gut microbiota composition toward the increase of beneficial organisms such as Blautia and Lactobacillus and a reduction in NF-κB expression. These findings support our hypothesis that cheeses might be explored as functional matrices for the efficacious delivery of probiotic strains to consumers.     

Effects of high pressure on proteolytic enzymes in cheese: relationship with the proteolysis of ewe milk cheese

Ewe milk cheeses were submitted to 200, 300, 400, and 500 MPa (2P to 5P) at 2 stages of ripening (after 1 and 15 d of manufacturing; P1 and P15). The high-pressure-treated cheeses showed a more important hydrolysis of β-casein than control and 2P1 cheeses. Degradation of α_s1-casein was more important in 3P1, 4P1, and P15 cheeses than control and 2P1 cheeses. The 5P1 cheeses exhibited the lowest degradation of α_s-caseins, probably as a consequence of the inactivation of residual chymosin. Treatment at 300 MPa applied on the first day of ripening increased the peptidolytic activity, accelerating the secondary proteolysis of cheeses. The 3P1 cheeses had extensive peptide degradation and the highest content of free amino acids. Treatments at 500 MPa, however, decelerated the proteolysis of cheeses due to a reduction of microbial population and inactivation of enzymes.     

Effect of various high-pressure treatments on the properties of reduced-fat Cheddar cheese

A major problem with reduced-fat cheese is the difficulty in attaining the characteristic flavor and texture of typical full-fat versions. Some previous studies have suggested that high hydrostatic pressure (HHP) can accelerate the ripening of full-fat cheeses. Our objective was to investigate the effect of HHP on reduced-fat (~7.3% fat) Cheddar cheese, with the goal of improving its flavor and texture. We used a central composite rotatable design with response surface methodology to study the effect of pressure and holding time on the rheological, physical, chemical, and microbial characteristics of reduced-fat Cheddar cheese. A 2-level factorial experimental design was chosen to study the effects of the independent variables (pressure and holding time). Pressures were varied from around 50 to 400 MPa and holding times ranged from 2.5 to 19.5 min. High pressure was applied 1 wk after cheese manufacture, and analyses were performed at 2 wk, and 1, 3, and 6 mo. The insoluble calcium content as a percentage of total Ca in cheeses were not affected by pressure treatment. Pressure applications ≥225 MPa resulted in softer cheese texture during ripening. Pressures ≥225 MPa increased melt, and resulted in higher maximum loss tangent values at 2 wk. Pressure treatment had a greater effect on cheese microbial and textural properties than holding time. High-pressure-treated cheeses also had higher pH values than the control. We did not observe any significant difference in rates of proteolysis between treatments. In conclusion, holding times of around 5 min and pressures of ≥225 MPa could potentially be used to improve the excessively firm texture of reduced-fat cheese.     

Probiotic effects of feeding heat-killed Lactobacillus acidophilus and Lactobacillus casei to Candida albicans-colonized immunodeficient mice

Probiotic bacteria can protect immunodeficient mice from orogastric candidiasis but cause some pathology of their own. Severely immunodeficient patients may be at risk if fed viable probiotics, so this study evaluated the probiotic potential of nonviable probiotic bacteria to protect immunodeficient mice from Candida albicans infections. Heat-killed probiotic bacteria were fed to gnotobiotic bg/bg-nu/nu and bg/bg-nu/+ mice to ascertain if they could protect the mice from mucosal and systemic candidiasis. Both heat-killed Lactobacillus acidophilus (HKLA) and heat-killed Lactobacillus casei (HKLC), in comparison to control mice not fed the probiotic bacteria but challenged (oral) with C. albicans, suppressed the severity of orogastric candidiasis in bg/bg-nu/nu mice at 2 weeks after colonization with C. albicans, inhibited disseminated candidiasis in C. albicans-colonized bg/bg-nu/+ mice at 4 weeks after colonization, and suppressed the number of viable C. albicans in the alimentary tract. HKLA, but not HKLC, treatment inhibited disseminated candidiasis in bg/bg-nu/nu mice at 2 weeks after oral challenge and enhanced the proliferative responses of splenocytes from C. albicans-colonized bg/bg-nu/+ mice to C. albicans antigens. Neither HKLA nor HKLC were able to prolong the survival of gnotobiotic bg/bg-nu/nu mice after oral challenge with C. albicans. These results demonstrate that heat-killed lactobacilli can induce some (limited) protection (probiotic effect) against candidiasis in mice.     

Amino acids evolution during ripening of goats' milk cheese manufactured with different coagulants

Powdered plant coagulant obtained from the cardoon flowers ( Cynara cardunculus ) was compared with calf rennet for the manufacture of traditional raw goats' milk cheese, by determining differences in the profiles of the amino acids throughout the ripening period. Derivatisation with o -phtaldialdehyde and a C₁₈ column were used for chromatographic separations. In order to establish the relationships between the different variables and to detect the most important causes of variability, principal component analysis was applied to the free amino acids data, which reduced to two dimensions where cheese samples from 2 to 60 days and cheeses from 90 to 120 days were distributed dependently of ripening time and coagulant used. Leu, Val, Lys, Gly, Tyr and Asp were correlated with PC1, which separated the samples according to their ripening time. Arg, His, Trp, Ser and Thr were correlated with PC2, which separated the samples according to the coagulant used.     

Prevention of gastrointestinal infection using immunobiological methods with milk fermented with Lactobacillus casei and Lactobacillus acidophilus

The protective effect of feeding milk fermented with a mixture of Lactobacillus casei sp. and Lb. acidophilus sp. against Salmonella typhimurium infection in mice was compared with that obtained feeding milks fermented with these microorganisms individually. The survival rate obtained after oral infection with Sal. typhimurium was 100% in mice pretreated by feeding during 8 d with the mixture of Lb. casei and Lb. acidophilus fermented milks. Similar treatments with the individual milks were ineffective. Moreover, mice became more susceptible to infection with Sal. typhimurium after such treatment. The colonization of liver and spleen with the pathogen was markedly inhibited by the pretreatment with the mixture of fermented milk, while such inhibition was not observed using the Lb. casei and Lb. acidophilus milks. The highest levels of anti-salmonellae antibodies in serum and in intestinal fluid were found in the group of mice fed with the mixture and with Lb. casei fermented milk respectively. However, this latter milk was not effective in protecting against Sal. typhimurium. When the mice were first infected with Sal. typhimurium and then fed with the mixture of fermented milks, pathogen colonization was not prevented. The results suggest that the augmentation of resistance to salmonellae caused by the treatment with Lb. casei- + Lb. acidophilus-fermented milk was due to the anti-salmonellae protective immunity mainly mediated by the mucosal tissue. Milk fermented with this mixture could be used as an immunobiological method to prevent gastrointestinal infection.     

嗜酸乳杆菌发酵生产低脂干酪凝乳工艺的优化

以脱脂乳为原料,采用嗜酸乳杆菌发酵进行预酸化,对其生产的低脂干酪凝乳工艺条件进行了研究。实验选取凝乳pH、氯化钙添加量、凝乳酶添加量、凝乳温度4个影响因素,以干酪产率、乳清中非脂乳固体物质残留量、嗜酸乳杆菌活菌数为指标,采用L₉（3⁴）正交实验进行优化。结果表明,凝乳的最佳工艺参数为凝乳pH6.0,氯化钙添加量0.02%（w/w）,凝乳酶添加量0.01%（w/w,酶活20000u/g）,凝乳温度35℃,以此条件生产的低脂干酪脂肪含量小于5%,干酪产率29.41%,乳清中非脂乳固体物质残留量5.66%,嗜酸乳杆菌活菌数在10⁹cfu/mL以上。     

嗜酸乳杆菌发酵干酪成熟过程中蛋白质降解产生的肽类的性质分析

对嗜酸乳杆菌和嗜热链球菌发酵生产的具有功能性的低脂和全脂干酪4℃成熟90d时蛋白质的降解程度进行了研究。通过电泳分析表明,两种干酪成熟后蛋白质的降解程度比市售的普通低脂和全脂干酪要充分,且低脂干酪比全脂干酪蛋白降解程度要深,降解物含量要多;反相高效液相色谱(RP-HPLC)显示的干酪中肽图谱表明,成熟后,虽然低脂干酪和全脂干酪中仍然存在和牛奶中相同的蛋白物质,但部分发生了降解并产生了新的肽类,且两种干酪所生成的肽组分各不相同。     

温度对嗜酸乳杆菌和干酪乳杆菌发酵羊奶的影响

以鲜羊奶为原料,保加利亚乳杆菌和嗜热乳链球菌为基础发酵剂,在此基础上分别添加嗜酸乳杆菌和干酪乳杆菌,研究温度对两菌株发酵羊奶过程中酸度、pH、活菌数及总活茵数的影响.结果表明:嗜酸乳杆菌酸羊奶的最佳发酵温度为37℃,此时凝乳时间为4h,酸度、pH、嗜酸乳杆菌数和总活菌数分别为97.8 0T、3.88、1.8×107 CFU/mL和1.38×109 CFU/mI,感官评价总分可达到8.35分;干酪乳杆菌羊奶的最佳发酵温度为39℃,此时酸羊奶凝固时间为4.5h,酸度、pH、干酪乳杆菌数和总茵数分别为79.2 °T、4.48、1.56×108 CFU/mL和1.81×109 CFU/mL,感官评价总分可达到7.00分.     

Probiotic white cheese with Lactobacillus acidophilus

The objective of the present study was to determine the effects of Lactobacillus acidophilus on the sensory attributes, ripening time, and composition of Turkish white cheese and to investigate the survival of L. acidophilus during ripening of the cheese stored in vacuum or in brine. Two types of white cheeses, traditional cheese (control, made with Lactococcus lactis ssp. lactis and Lactococcus lactis ssp. cremoris) and probiotic cheese (made with Lactococcus lactis ssp. lactis, Lactococcus lactis ssp. cremoris and L. acidophilus 593 N), were produced and ripened in vacuum pack or in brine at 4°C for 90 days. Cheese samples were assessed for microbiological and compositional properties, proteolysis, and sensory evaluation at different ripening stages. On ripening in vacuum pack, L. acidophilus survived to numbers >10⁷ cfu g⁻¹, which is necessary for positive effects on health. Protein, dry matter, salt content, and percentage of lactic acid in the vacuum-packed and brine-salted probiotic cheeses were significantly different. Also, the lactic acid content of probiotic cheeses was slightly higher than that of the controls for both vacuum- and brine-packed cheeses. Vacuum-packed probiotic cheese had the highest levels of proteolysis and the highest sensory scores of all cheeses. Consequently, L. acidophilus could be used for the manufacturing of probiotic white cheese to shorten ripening time and vacuum packaging is the preferred storage format.     

Lysis of Lactobacillus casei 5Mn 373 accelerates Grana cheese ripening

The influence of the adjunct of a peptidolytic Lactobacillus casei strain on Grana cheese ripening was studied. Strain erythromycin resistance enabled the monitoring of its growth and death kinetics during cheese maturation. Cell lysis was estimated by the dosage of intracellular X-prolyl-dipeptidyl aminopetidase in cheese extracts. L. casei growth reached a maximum level after the second month of cheese ripening when the initial added cell level was 5×10⁵ cfu/g cheese, while L. casei counts decreased from the beginning of the ripening period when the initial added cell level was 4.5×10⁷ cfu/g cheese. The maximum death rate occurred two months after the maximal cell growth, and bacterial lysis was observed approximately two months later. The characteristic amino acid pattern of control Grana cheese was obtained for all of the mature experimental cheeses independently of the inoculum size, and more rapidly when higher amounts of inocula were used due to L. casei cell lysis. The adjunct of the L. casei strain to cheese-milk substantially shortened the ripening time with no negative effects on cheese-making, chemical gross composition or flavour in the mature experimental cheeses compared to the control.     

Proteolysis and biogenic amine buildup in high-pressure treated ovine milk blue-veined cheese

Penicillium roqueforti plays an important role in the ripening of blue-veined cheeses, mostly due to lactic acid consumption and to its extracellular enzymes. The strong activity of P. roqueforti proteinases may bring about cheese over-ripening. Also, free amino acids at high concentrations serve as substrates for biogenic amine formation. Both facts result in shorter product shelf-life. To prevent over-ripening and buildup of biogenic amines, blue-veined cheeses made from pasteurized ovine milk were high-pressure treated at 400 or 600 MPa after 3, 6, or 9 wk of ripening. Primary and secondary proteolysis, biogenic amines, and sensory characteristics of pressurized and control cheeses were monitored for a 90-d ripening period, followed by a 270-d refrigerated storage period. On d 90, treatments at 400 MPa had lowered counts of lactic acid bacteria and P. roqueforti by less than 2 log units, whereas treatments at 600 MPa had reduced lactic acid bacteria counts by more than 4 log units and P. roqueforti counts by more than 6 log units. No residual α-casein (CN) or κ-CN were detected in control cheese on d 90. Concentrations of β-CN, para-κ-CN, and γ-CN were generally higher in 600 MPa cheeses than in the rest. From d 90 onwards, hydrophilic peptides were at similar levels in pressurized and control cheeses, but hydrophobic peptides and the hydrophobic-to-hydrophilic peptide ratio were at higher levels in pressurized cheeses than in control cheese. Aminopeptidase activity, overall proteolysis, and free amino acid contents were generally higher in control cheese than in pressurized cheeses, particularly if treated at 600 MPa. Tyramine concentration was lower in pressurized cheeses, but tryptamine, phenylethylamine, and putrescine contents were higher in some of the pressurized cheeses than in control cheese. Differences in sensory characteristics between pressurized and control cheeses were generally negligible, with the only exception of treatment at high pressure level (600 MPa) at an early ripening stage (3 wk), which affected biochemical changes and sensory characteristics.     

Probiotic in lamb rennet paste enhances rennet lipolytic activity, and conjugated linoleic acid and linoleic acid content in Pecorino cheese

Cheeses manufactured using traditional lamb rennet paste, lamb rennet paste containing Lactobacillus acidophilus, and lamb rennet paste containing a mix of Bifidobacterium lactis and Bifidobacterium longum were characterized for the lipolytic pattern during ripening. Lipase activity of lamb rennet paste, lamb rennet containing Lb. acidophilus, and lamb rennet containing a mix of bifidobacteria was measured in sheep milk cream substrate. Rennet paste containing probiotics showed a lipase activity 2-fold greater than that displayed by traditional rennet. Total free fatty acid (FFA) in sheep milk cream was lower in lamb rennet paste (981 μg/g of milk cream) than in lamb rennet containing Lb. acidophilus (1,382.4 μg/g of milk cream) and in lamb rennet containing a mix of bifidobacteria (1,227.5 μg/g of milk cream) according to lipase activity of lamb rennet paste. The major increase of FFA in all cheeses occurred during the first 30 d of ripening with the greatest values being observed for C16:0, C18:0 C18:1. At 60 d of ripening all cheeses showed a reduction in the amount of free fatty acids; in particular, total free fatty acids underwent a decrease of more than 30% from 30 to 60 d in cheeses manufactured using traditional lamb rennet paste, whereas the same parameter decreased 10% in cheeses manufactured using lamb rennet paste containing Lb. acidophilus and cheeses manufactured using lamb rennet paste containing a mix of B. lactis and B. longum. Cheese containing Lb. acidophilus was characterized by the greatest levels of total conjugated linoleic acids (CLA) 9-cis, 11-trans CLA and 9-trans, 11-trans CLA, whereas cheese containing bifidobacteria displayed the greatest levels of free linoleic acid. Rennet pastes containing viable cells of Lb. acidophilus and a mix of B. lactis and B. longum were able to influence the amount of FFA and CLA in Pecorino cheese during ripening.