Volatile composition and improvement of the aroma of industrial Manchego cheese by using Lactobacillus paracasei subsp. paracasei as adjunct and other autochthonous strains as starters

The use of several autochthonous strains of lactic acid bacteria, including Lactobacillus paracasei subsp. paracasei as adjunct of the starter in the manufacture of Manchego cheese, was evaluated in an attempt to improve the aroma of the industrial Manchego cheese. Volatile composition and odour characteristics were evaluated and compared to those in Manchego cheese manufactured with a commercial starter (CS) culture and with raw milk cheese manufactured without starter. Manchego cheeses manufactured with two autochthonous strains of Lactococcus lactis subsp. lactis displayed a similar volatile profile and odour characteristics to the cheese made with the CS. The use of the strain Lactobacillus paracasei subsp. paracasei CECT 7882 as adjunct of the Lactococcus strains produced cheeses with higher amounts of some free fatty acids and alcohols, acetoin, lactones, phenylacetaldehyde, 2-phenylethanol and linalool, and higher scores of the odour intensity, odour quality, and ewe’s milk odour than the CS cheeses. It resulted in an intensification and improvement of industrial Manchego cheese aroma.     

Use of Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris isolated from TRFM in manufacturing of functional low-fat cheeses

Abstract: The purpose of this study was to manufacture new functional low‐fat cheeses using Taiwanese ropy fermented milk (TRFM) and Lactococcus lactis subsp. cremoris strains isolated from TRFM. After 28 d of ripening and storage, the viable populations of lactic acid bacteria (LAB) in the low‐fat cheeses made with L. lactis subsp. cremoris TL1 (TL1), L. lactis subsp. cremoris TL4 (TL4), and TRFM still maintained above 10⁸ CFU/g. The low‐fat cheeses made with TL1 and TRFM showed higher moisture contents than the cheeses made with TL4, full‐fat, and low‐fat cheese controls. The low‐fat cheeses made with TL1 and TL4 had higher customer preferential scores similar to full‐fat cheese control in the sensory evaluation. Additionally, the low‐fat cheeses fermented with TL1, TL4, and TRFM for 4 h had higher 1,1‐diphenyl‐2‐picrylhydrazyl (DPPH) free radical‐scavenging and ferrous ion‐chelating abilities than the cheeses fermented with the starters for 8 h, full‐fat, and low‐fat cheese controls. A better angiotensin‐converting enzyme (ACE) inhibition activity was also observed in the low‐fat cheeses made with TL1, TL4, and TRFM than that in the full‐fat and low‐fat cheese controls during ripening and storage period. Practical Application: As health‐conscious consumers continue to seek low‐fat alternatives in their diets, there remain strong interests for the dairy industry to develop low‐fat cheeses to meet the demands. This study clearly demonstrated that the low‐fat cheeses fermented with TL1 for 4 h showed a better overall acceptability and possessed antioxidative abilities and ACE inhibitory activities than other cheeses tested in this study. By improving its flavor and investigating the possible mechanisms of its functionalities in the future, this low‐fat cheese might possibly be commercialized and give a positive impact on cheese consumption in the future.     

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.     

Manufacture of Turkish Beyaz cheese added with probiotic strains

The survival of the probiotic strains Lactobacillus fermentum (AB5-18 and AK4-120) and Lactobacillus plantarum (AB16-65 and AC18-82), all derived from human faces, was investigated in Turkish Beyaz cheese production. Three batches of Turkish Beyaz cheese were produced: one with the test probiotic culture mix (P), another with a commercial starter culture mix including Lactoccocus lactis subsp. cremoris, Lactococcus lactis subsp. lactis (C) and the third with equal parts of the commercial starter culture mix and test probiotic culture mix (CP). The cheeses were ripened at 4 °C for 120 days and the viability of cultures was determined monthly. Cheese samples were analyzed for total solids, fat in solids, titratable acidity, pH, salt in total solids, proteolysis, sensory evaluation, aroma compounds and biogenic amines. While initial lactic acid bacteria load in P cheese was 2.7 × 10⁹ at the beginning, it was 7.42 × 10⁷ cfu/g at the end of 120 days of ripening. The results showed that test probiotic culture mix was successfully used in cheese production without adversely affecting the cheese quality during ripening. The chemical composition and sensory quality of P cheeses were also comparable with C cheeses. The present study indicates that probiotic cultures of human origin are feasible for Turkish Beyaz cheese production.     

Influence of a defined-strain starter and Lactobacillus plantarum as adjunct culture on volatile compounds and sensory characteristics of Manchego cheese

Three batches of Manchego cheese were manufactured using one of the following starter culture systems: (1) a defined strain starter culture comprising Lactococcus lactis subsp. lactis and Leuconostoc mesenteroides subsp. dextranicum; (2) the above-defined strain starter culture and an adjunct culture (Lactobacillus plantarum), all these strains being isolated from high-quality Manchego cheeses and (3) a commercial starter consisting of two strains of Lactococcus lactis. Differences in volatile profile and the sensory characteristics of these cheeses were studied. After 4 months of ripening, the two batches of cheese made with the defined strain starter cultures obtained the highest scores for sensory attributes and for the overall impression. Additionally, Purge & Trap and SDE analysis showed a more complex volatile profile in these cheeses than in those made with the commercial starter. Extending the maturation time to 8 months for cheeses made with the defined starter cultures led to significant higher levels of free fatty acids and ethyl esters in those cheeses made without adjunct culture. However, panelists did not find significant differences among the sensory characteristics of the two cheeses.     

Microbial diversity and dynamics of Pecorino di Filiano PDO,a traditional cheese of Basilicata region (Southern Italy)

The evolution of microbial populations of ‘Pecorino di Filiano’ (PF) cheese was investigated during ripening in natural cave and storeroom. 62.5% of isolates grow at 45 and 15 °C and 77.7% showed high salt concentrations tolerance. Brevibacterium linens was dominant in surface samples. Lactobacillus delbrueckii subsp. bulgaricus and Lactobacillus paracasei subsp. paracasei were more frequently isolated both surface and core samples, while Leuconostoc lactis and Leuconostoc mesenteroides subsp. mesenteroides prevailed among Leuconostoc isolates. Our results suggest the importance of the ripening environment of cheeses and how a biological ecosystem affects and produces the typical features of artisanal products.     

Impact of native Lactobacillus paracasei subsp. paracasei and Pediococcus spp. as adjunct cultures on sensory quality of Iranian white brined cheese

Paired wild‐type cultures consisting of a Lactobacillus paracasei subsp. paracasei (three strains) or Pediococcus pentosaceus (one strain) and a Pediococcus inopinatus (five strains) were used as adjunct cultures in the production of Iranian white brined cheese. After 8 weeks of ripening, adjunct‐treated cheeses produced by L. paracasei subsp. paracasei and P. inopinatus received significantly higher scores for flavour/taste, aroma, texture and overall preference than those produced by P. pentosaceus and P. inopinatus as well as the control cheese (P < 0.05). In conclusion, a greater improvement of sensory quality of cheeses was strongly associated with the presence of L. paracasei subsp. paracasei rather than pediococci.     

Proteolytic properties of Turkish white‐brined cheese (Beyaz peynir) made by using wild‐type Lactococcal strains

The development of proteolysis in white‐brined Turkish cheese made by using wild strains of Lactococcus lactis subsp. lactis (namely MBLL9, MBLL23 and MBL27) was monitored for 90 days. Proteolysis in cheeses was investigated using urea‐PAGE gel electrophoresis of pH 4.6‐insoluble and RP‐HPLC of both 70% ethanol‐insoluble and 70% ethanol‐soluble nitrogen fractions. Results indicated that developments of proteolysis in the experimental cheeses were strain dependent. The degradation of casein fractions was more evident in the cheeses made using strain MBLL23. The lowest levels of proteolysis and development of acidity were obtained in the cheese made using strain MBLL9.     

The effects of incorporating wild‐type strains of Lactococcus lactis into Turkish white‐brined cheese (Beyaz peynir) on the fatty acid and volatile content

The development of free fatty acids (FFA) and volatile flavour compounds in the Turkish white‐brined cheese Beyaz peynir made by using three wild strains of Lactococcus lactis subsp. lactis was investigated over 90 days. Results showed that production of both FFA and flavour compounds in the control (PK1) and experimental cheeses (MBLL9, MBLL23 and MBL27) was strain dependent. The hydrolysis of milk fat was more evident in the cheese made using Lc. lactis subsp. lactis MBL27. Considering the production of fat breakdown compounds and acidification activities of the strains MBLL23 and MBL27, the combination of these strains could be proposed for the production of white‐brined cheese.     

Proteolytic activity of three probiotic strains in semi-hard cheese as single and mixed cultures: Lactobacillus acidophilus,Lactobacillus paracasei and Bifidobacterium lactis

The influence of three probiotic strains (Lactobacillus acidophilus, Lactobacillus paracasei and Bifidobacterium lactis) in semi-hard cheese proteolysis patterns was assessed. Probiotics were inoculated both as single cultures and as a three-strain mix, and added to milk either after a pre-incubation step or directly to the vat. B. lactis did not show any effect on proteolysis of cheeses, while L. paracasei showed limited impact at the end of the ripening. In contrast, L. acidophilus significantly influenced secondary proteolysis from the beginning of ripening, causing an increase in the levels of small nitrogen-containing compounds and free amino acids and changes in the peptide profiles. The effect of Lactobacillus acidophilus on peptidolysis was more noticeable when it was added to cheese–milk after pre-incubation in an enriched milk fat substrate. Similar results obtained with the three-strain mixed culture, suggesting that L. acidophilus played a major role in secondary proteolysis of probiotic cheeses in this trial.     

Influence of microfiltration and adjunct culture on quality of Domiati cheese

The effects of microfiltration and pasteurization processes on proteolysis, lipolysis, and flavor development in Domiati cheese during 2 mo of pickling were studied. Cultures of starter lactic acid bacteria isolated from Egyptian dairy products were evaluated in experimental Domiati cheese for flavor development capabilities. In the first trial, raw skim milk was microfiltered and then the protein:fat ratio was standardized using pasteurized cream. Pasteurized milk with same protein:fat ratio was also used in the second trial. The chemical composition of cheeses seemed to be affected by milk treatment—microfiltration or pasteurization—rather than by the culture types. The moisture content was higher and the pH was lower in pasteurized milk cheeses than in microfiltered milk cheeses at d 1 of manufacture. Chemical composition of experimental cheeses was within the legal limits for Domiati cheese in Egypt. Proteolysis and lipolysis during cheese pickling were lower in microfiltered milk cheeses compared with pasteurized milk cheeses. Highly significant variations in free amino acids, free fatty acids, and sensory evaluation were found among the cultures used in Domiati cheesemaking. The cheese made using adjunct culture containing Lactobacillus delbrueckii ssp. lactis, Lactobacillus paracasei ssp. paracasei, Lactobacillus casei, Lactobacillus plantarum, and Enterococcus faecium received high scores in flavor acceptability. Cheeses made from microfiltered milk received a higher score in body and texture compared with cheeses made from pasteurized milk.     

Lactic acid bacteria with cholesterol-lowering properties for dairy applications: In vitro and in situ activity

Cholesterol-lowering activity is one of the most promising properties of lactic acid bacteria with probiotic characteristics. In the present study, 58 potentially probiotic lactic acid bacteria were tested for their ability to survive in vitro digestion and reduce cholesterol in a medium containing cholesterol and bile acids. The best-performing strains (Lactobacillus casei VC199, Lactobacillus paracasei ssp. paracasei SE160 and VC213, Lactobacillus plantarum VS166 and VS513, Enterococcus faecium VC223, and Enterococcus lactis BT161) resulted in a 42 to 55% reduction of the cholesterol level in broth and were further tested in cheese manufacture. The cholesterol content in all the cheeses decreased with ripening. All the strains were present in the cheese at levels higher than 10⁷ cfu/g until 60 d of ripening, the highest reductions (up to 23%) being obtained when Lb. paracasei ssp. paracasei VC213 and E. lactis BT161 were added during the cheese-making. The adjunct cultures had no negative effect on the sensory characteristics of the cheese. Thus, these strains with proven in vitro properties are good candidates for novel probiotic-containing formulations and could be used to functionalize foods such as dairy fermented products.     

Technological and probiotic role of adjunct cultures of non-starter lactobacilli in soft cheeses

The influence of two cheese-isolated Lactobacillus strains on cheese composition, acceptability and probiotic capacity was assessed. Soft cheeses with and without the addition of Lactobacillus plantarum I91 or Lactobacillus paracasei I90 were prepared. Gross composition was assessed and secondary proteolysis was described by soluble fractions and free amino acids profiles. Acceptability was determined by a panel of 98 non-trained consumers. Cheeses harboring added Lactobacillus strains were also studied in vivo to evaluate their probiotic capacity. Gross composition of the cheeses was similar for control and treated (Lactobacillus-added) cheeses. Peptidolysis increased in cheeses with added lactobacilli, which was evidenced by a higher free amino acid content. Overall, the acceptability of the cheeses was good: 65%–80% of the consumers said that they “liked very much” or “liked” the cheeses. Cheeses with L. plantarum I91 showed the highest changes in composition and proteolysis and were the most accepted ones. On the contrary, composition of cheeses with L. paracasei I90 was similar to that of the controls, but these samples were less accepted than cheeses without lactobacilli. The oral administration of cheese containing L. plantarum I91 or L. paracasei I90 proved to be safe and able to enhance the number of IgA + cells in the small intestine lamina propria of mice. The use of selected strains of NSLAB exerted a technological and probiotic role: it contributed to the standardization of cheese quality and induced benefic health effects at the gut mucosa in vivo.     

Enzymatic activity of lactic acid bacteria (with antimicrobial properties) isolated from a traditional Spanish cheese

Twenty-four strains of lactic acid bacteria (LAB) isolated from a traditional Spanish cheese (Genestoso cheese) were evaluated for their enzymatic activities (acidifying and proteolytic abilities and carboxypeptidase, aminopeptidase, dipeptidase, caseinolytic and esterase activities), in order to select indigenous strains of technical interest for the manufacture of cheese. These strains were selected on the basis of their antimicrobial activity relative to five reference strains and were identified as Lactococcus lactis subsp. lactis (thirteen strains), Leuconostoc mesenteroides (two strains), Leuconostoc pseudomesenteroides (one strain), Lactobacillus paracasei (two strains), Lactobacillus plantarum (one strain) and Enterococcus faecalis (five strains).     

The effect of ripening and storage conditions on the distribution of tyramine,putrescine and cadaverine in Edam-cheese

The aim of the work was to describe the development of selected biogenic amines (histamine, tyramine, putrescine and cadaverine) in 4 layers of Dutch-type cheese (Edam-cheese) depending on 3 ripening/storage regimes during a 98-day period. Biogenic amines were analysed by means of ion-exchange chromatography. A further goal was to identify microbial sources of biogenic amines in the material analysed. Phenotype characterization and repetitive sequence-based PCR fingerprinting were used to identify the isolated bacteria. The highest content of tyramine, putrescine and cadaverine was determined in cheeses stored in a ripening cellar at a temperature of 10 °C during the whole observation period. Lower biogenic amines content was determined in samples which were moved into a cold storage device (5 °C) after 38 days of storage in a ripening cellar (10 °C). The lowest concentrations of biogenic amines were detected in cheeses which were moved into a cold storage device (5 °C) after 23 days of storage in a ripening cellar (10 °C). During the 98-day period, histamine was not detected in any of the regimes. Within the cheeses analysed, non-starter lactic acid bacteria Lactobacillus curvatus, Lactobacillus casei/paracasei and Lactobacillus plantarum were detected as the main producers of the biogenic amines tested. In starter bacteria Lactococcus lactis subsp. lactis and Lactococcus lactis subsp. cremoris the decarboxylase activity tested was not detected.     

In vitro evaluation of physiological probiotic properties of different lactic acid bacteria strains of dairy and human origin

Eleven lactic acid bacteria strains of importance to the dairy industry were subjected to in vitro analyses to determine their probiotic potential. Seven strains were isolated from ewe’s and cow’s milk (Enterococcus faecalis – five –, Lactococcus lactis and Lactobacillus paracasei). Four were obtained from American Type Culture Collection (ATCC), isolated from cheese (Lactobacillus casei 393), human feces (L. paracasei 27092 and Lactobacillus rhamnosus 53103) and used in cheese making (L. lactis 54104). Although none of the strains was able to degrade mucin, all E. faecalis showed, at least, one transferable antibiotic resistance, which excluded them as candidates for addition to foods. Of the remaining six safe strains, L. lactis strains were more tolerant to low pH than Lactobacillus spp.; all were tolerant to pancreatin and bile salts and showed antibacterial activity. The highest level of adhesion to Caco-2 cells was observed with L. lactis 660, even higher than L. rhamnosus ATCC 53103 (recognized probiotic and used as control). The physiological probiotic properties of these strains, mainly isolated from dairy sources, are interesting in view of their use in cheese productions as starter and non starter cultures. The five LAB safe strains studied may have potential as novel probiotics in the dairy foods.     

Quantitative PCR for the specific quantification of Lactococcus lactis and Lactobacillus paracasei and its interest for Lactococcus lactis in cheese samples

The first objective of this work was to develop real-time quantitative PCR (qPCR) assays to quantify two species of mesophilic lactic acid bacteria technologically active in food fermentation, including cheese making: Lactococcus lactis and Lactobacillus paracasei. The second objective was to compare qPCR and plate counts of these two species in cheese samples. Newly designed primers efficiently amplified a region of the tuf gene from the target species. Sixty-three DNA samples from twenty different bacterial species, phylogenetically related or commonly found in raw milk and dairy products, were selected as positive and negative controls. Target DNA was successfully amplified showing a single peak on the amplicon melting curve; non-target DNA was not amplified. Quantification was linear over 5 log units (R² > 0.990), down to 22 gene copies/μL per well for Lc. lactis and 73 gene copies/μL per well for Lb. paracasei. qPCR efficiency ranged from 82.9% to 93.7% for Lc. lactis and from 81.1% to 99.5% for Lb. paracasei. At two stages of growth, Lc. lactis was quantified in 12 soft cheeses and Lb. paracasei in 24 hard cooked cheeses. qPCR proved to be useful for quantifying Lc. lactis, but not Lb. paracasei.     

Biogenic amine content and proteolysis in Manchego cheese manufactured with Lactobacillus paracasei subsp. paracasei as adjunct and other autochthonous strains as starters

Two different autochthonous strain starter cultures, in which the acidifying starter was composed of strains of Lactococcus lactis, were used for the manufacture of pasteurised milk Manchego cheese. Proteolysis parameters, biogenic amines and sensory characteristics were evaluated and compared with those of commercial starter Manchego cheese and raw milk Manchego cheese manufactured without starter. Autochthonous starter cheeses, and especially those including Lactobacillus paracasei subsp. paracasei as adjunct, presented higher levels of proteolysis than in commercial starter cheese. The concentrations of total biogenic amines in autochthonous starter cheeses were much lower than in raw milk cheese and even lower than in commercial starter cheese. Cheese manufactured with the adjunct strain gave the best results for both flavour intensity and flavour quality, and was the most preferred by panellists. The results suggest that the culture containing Lb. paracasei subsp. paracasei as adjunct could be used for the manufacture of industrial Manchego cheese.     

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.     

Viability of Probiotics in Goat Cheese During Storage and Under Simulated Gastrointestinal Conditions

The objective of this study was to evaluate the suitability of two strains of probiotic bacteria (Bifidobacterium animalis subsp. lactis and Lactobacillus rhamnosus) incorporated individually into Boursin-type goat cheese and their in vitro resistance through the passage of the gastrointestinal tract in the cheese. The viability of B. lactis and Lb. rhamnosus cultures was unaffected throughout 35 days of storage at 4 °C, with a final count of >?7 log CFU/g. No significant difference (p?>?0.05) was observed between probiotic treatments and control in relation to pH and titratable acidity. B. animalis presented greater resistance to the artificial gastric and enteric juices than Lb. rhamnosus, with mean decreases in the initial populations of 0.2 and 4.0 log CFU/g within 35 days of storage, respectively. The addition of probiotic cultures did not affect the consumer acceptance of the goat cheeses. Three segments of consumers with different liking were identified. The results demonstrated that “Boursin” goat cheese is a promising matrix for the incorporation and protection of B. animalis subsp. lactis.