Effects of milk treatment with dynamic high pressure on microbial populations, and lipolytic and proteolytic profiles of Crescenza cheese

The effects of some physical treatments of milk (pasteurization and homogenization at 1000 bar) on the evolution of the microbial populations, as well as on the proteolytic and lipolytic profiles, of Crescenza, a traditional Italian soft cheese, were studied in relation to its shelf-life. The dynamic, high-pressure homogenization (HPH) treatment of milk caused a reduction in the cell loads attained by the expiry date of yeasts and Pseudomonas spp. Moreover, the thermal and HPH treatments had different selective actions on the yeast populations, the former favouring oxidative species. In addition, the Crescenza obtained using milk treated at 1000 bar showed evidence of early and significant lipolysis.     

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7 mo of ripening showed a higher rate of pH decrease and lower final pH value compared with reactivation of samples withdrawn from the remaining mass of the cheese without fruit pieces, from cheese with free L. casei, and rennet cheese.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。     

Comparative evaluation of yogurt and low-fat cheddar cheese as delivery media for probiotic Lactobacillus casei

This study used Lactobacillus casei 334e, an erythromycin-resistant derivative of ATCC 334, as a model to evaluate viability and acid resistance of probiotic L. casei in low-fat Cheddar cheese and yogurt. Cheese and yogurt were made by standard methods and the probiotic L. casei adjunct was added at approximately 10(7) CFU/g with the starter cultures. Low-fat cheese and yogurt samples were stored at 8 and 2 degrees C, respectively, and numbers of the L. casei adjunct were periodically determined by plating on MRS agar that contained 5 microg/mL of erythromycin. L. casei 334e counts in cheese and yogurt remained at 10(7) CFU/g over 3 mo and 3 wk, respectively, indicating good survival in both products. Acid challenge studies in 8.7 mM phosphoric acid (pH 2) at 37 degrees C showed numbers of L. casei 334e in yogurt dropped from 10(7) CFU/g to less than 10(1) CFU/g after 30 min, while counts in cheese samples dropped from 10(7) CFU/g to about 10(5) after 30 min, and remained near 10(4) CFU/g after 120 min. As a whole, these data showed that low-fat Cheddar cheese is a viable delivery food for probiotic L. casei because it allowed for good survival during storage and helped protect cells against the very low pH that will be encountered during stomach transit.     

High-pressure homogenization of raw and pasteurized milk modifies the yield, composition, and texture of queso fresco cheese

High-pressure homogenization (HPH) of milk was studied as an alternative processing operation in the manufacturing of queso fresco cheese. Raw and pasteurized (65°C for 30 min) milks were subjected to HPH at 0, 100, 200, and 300 MPa and then used to manufacture queso fresco. The cheeses were evaluated for yield, moisture content, titratable acidity, nitrogen content, whey protein content, yield force, yield strain, and tactile texture by instrumental or trained panel analyses. The combination of HPH and thermal processing of milk resulted in cheeses with increased yield and moisture content. The net amount of protein transferred to the cheese per kilogram of milk remained constant for all treatments except raw milk processed at 300 MPa. The highest cheese yield, moisture content, and crumbliness were obtained for thermally processed milk subjected to HPH at 300 MPa. The principal component analysis of all measured variables showed that the variables yield, moisture content, and crumbliness were strongly correlated to each other and negatively correlated to the variables yield strain, protein content (wet basis), and sensory cohesiveness. It is suggested that the combination of thermal processing and HPH promotes thermally induced denaturation of whey protein, together with homogenization-induced dissociation of casein micelles. The combined effect results in queso fresco containing a thin casein-whey matrix that is able to better retain sweet whey. These results indicate that HPH has a strong potential for the manufacture of queso fresco with excellent yield and textural properties.     

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.     

Camembert-type cheese quality and safety implications in relation to the timing of high-pressure processing during aging

Bloomy rind cheeses, including Brie, Camembert, and related varieties, are at high risk of contamination by environmental pathogens during manufacture and ripening. This risk is particularly high during ripening due to open-air exposure of the product. Currently, no kill step is applied after manufacture or post ripening to control food safety risks associated with Listeria monocytogenes contamination. Instead, cheesemakers must rely on sanitation and environmental monitoring to reduce this risk. High-pressure processing (HPP) is a nonthermal food-processing technology that can effectively reduce bacterial contaminants with minimal impact on the organoleptic properties of various foods. The objective of this study was to evaluate HPP as a potential intervention to maintain Camembert cheese quality and reduce risk associated with L. monocytogenes. Timing of HPP treatments (3, 11, and 45 d after manufacture) was based on the growth of L. monocytogenes during Camembert cheese ripening. High-pressure processing treatment of fully ripened cheeses (45 d) resulted in destruction of the surface mold, which caused browning and yellowing of the cheese rind. Applying HPP treatment earlier in the ripening process (11 d) resulted in a similar degradation of cheese appearance, which did not improve with continued ripening. Applying HPP treatment shortly after production (3 d; before the surface flora developed) delayed the development of the cheese rind and the textural ripening of the cheese. This early treatment time also resulted in free whey being expelled from the cheese, creating a firmer body. Applying HPP 11 d after manufacture resulted in >5 log reduction of L. monocytogenes at 450 and 550 MPa with holding times of 10 min. Although HPP was effective at reducing L. monocytogenes associated with bloomy rind cheeses, the quality deterioration would be unacceptable to consumers. Cheesemakers must continue to emphasize sanitation and environmental monitoring to reduce the risk of L. monocytogenes in bloomy rind cheeses.     

Production of low-fat Chanco cheese using homogenized milk and adjunct lactic culture

The objective of this study was to determine the effect of the partial homogenization of milk and the use of an adjunct lactic culture on the processing, quality and yield of low-fat Chanco cheese.
  Treatments were: T1 normal-fat control and T2 low-fat control. The low-fat treatments were: T3 using homogenized milk, T4 with adjunct lactic culture and T5 both variables.
   It was concluded that the homogenization slightly increased the time of rest after cutting of the curd, and the addition of adjunct lactic culture increased the acidity of whey during a second agitation. Nevertheless, the total time and pH at the end of the process were normal and without significant statistical difference ( P  > 0.05) between treatments. When using homogenized milk the losses of fat in whey decreased by 50% with respect to the low-fat control and nearly 80% in relation to the full fat control. Consequently, the yield was increased. The physical and chemical characteristics of the Chanco cheese were not affected by the studied variables. With respect to the sensory evaluation, significant differences in colour and aroma did not appear, but there were statistically significant differences (P <  0.05) in taste, hardness, adhesiveness, cohesiveness and elasticity between the full-fat control and all the low-fat treatments. In the case of general acceptance, the treatment with adjunct lactic culture was similar to the full-fat control. In addition, it showed the least difference (mild) in elasticity and taste in relation to this control .     

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.     

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.     

动态高压处理对牛乳中生物活性物质的影响

牛乳中含有乳铁蛋白、免疫球蛋白等多种有益身体健康的生物活性成分,在加工过程中如何降低其损失一直是重点关注的方向。动态高压处理技术又称超高压均质(UHPH),是一种较为新颖的加工技术手段之一。文章从美拉德反应产物、乳铁蛋白和免疫球蛋白等生物活性成分保留以及维生素损失等多个维度,总结了超高压均质在牛乳加工过程中的热负荷强度和关键营养成分保留方面的相关研究进展,并对超高压均质在乳制品加工领域的发展方向进行了展望。     

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。