Cheese Starter Research Articles

Исследование свойств производственных штаммов Lb. plantarum с целью оценки возможности их использования в составе заквасок для сыроделия

Исследование свойств производственных штаммов Lb. plantarum с целью оценки возможности их использования в составе заквасок для сыроделия

The Cheese Matrix Modulates the Immunomodulatory Properties of Propionibacterium freudenreichii CIRM-BIA 129 in Healthy Piglets.

Propionibacterium freudenreichii is a beneficial bacterium, used as a cheese starter, which presents versatile probiotic properties. These properties are strain-dependent. We hypothesized they may also be delivery vehicle-dependent. In this study, we thus explored in healthy piglets how the cheese matrix affects the immunomodulatory properties of P. freudenreichii. During 2 weeks, three groups of weaned piglets consumed, respectively, P. freudenreichii as a liquid culture (PF-culture), P. freudenreichii under the form of a cheese (PF-cheese), or a control sterile cheese matrix (Cheese-matrix). The in vivo metabolic activity of P. freudenreichii was assessed by determining short chain fatty acids (SCFA) concentration and bifidobacteria population in feces. Whatever the delivery vehicle, P. freudenreichii was metabolically active in piglets’ colon and enhanced both bifidobacteria and SCFA in feces. P. freudenreichii consumption decreased the secretion of TNFα and of IL-10 by peripheral blood mononuclear cells (PBMC). It did not alter IL-10, IFNγ, IL-17, and TNFα secretion in mesenteric lymph node immune cells (MLNC). PF-cheese enhanced significantly Treg phenotype, while PF-culture decreased significantly Th17 phenotype in PBMC and MLNC. Remarkably, only PF-cheese induced an increase of Th2 phenotype in PBMC and MLNC. Ex vivo stimulation of PBMC and MLNC by Lipopolysaccharides and Concanavalin A emphasized the difference in the immunomodulatory responses between PF-culture and PF-cheese group, as well as between PBMC and MLNC. This study shows the importance to consider the delivery vehicle for probiotic administration. It confirms the anti-inflammatory potential of P. freudenreichii. It opens new perspectives for the use propionibacteria-fermented products as preventive agents for inflammatory bowel diseases and intestinal infectious diseases.

Genome analysis of lactic acid bacterial strains selected as potential starters for traditional Slovakian bryndza cheese.

Genomes of 21 strains of lactic acid bacteria isolated from Slovakian traditional cheeses were sequenced on an Illumina MiSeq platform. Subsequently, they were analysed regarding taxonomic classification, presence of genes encoding defence systems, antibiotic resistance and production of biogenic amines. Thirteen strains were found to carry genes encoding at least one bacteriocin, 18 carried genes encoding at least one restriction-modification system, all strains carried 1-6 prophages and 9 strains had CRISPR-Cas systems. CRISPR-Cas type II-A was the most common, containing 0-24 spacers. Only 10% spacers were found to be homological to known bacteriophage or plasmid sequences in databases. Two Enterococcus faecium strains and a Lactococcus lactis strain carried antibiotic resistance genes. Genes encoding for ornithine decarboxylase were detected in four strains and genes encoding for agmatine deiminase were detected in four strains. Lactobacillus paraplantarum 251 L appeared to be the most interesting strain, as it contained genes encoding for two bacteriocins, a restriction-modification system, two CRISPR-Cas systems, four prophages and no genes connected with antibiotic resistance or production of biogenic amines.

Genome analysis of lactic acid bacterial strains selected as potential starters for traditional Slovakian bryndza cheese

ABSTRACT Genomes of 21 strains of lactic acid bacteria isolated from Slovakian traditional cheeses were sequenced on an Illumina MiSeq platform. Subsequently, they were analysed regarding taxonomic classification, presence of genes encoding defence systems, antibiotic resistance and production of biogenic amines. Thirteen strains were found to carry genes encoding at least one bacteriocin, 18 carried genes encoding at least one restriction–modification system, all strains carried 1–6 prophages and 9 strains had CRISPR-Cas systems. CRISPR-Cas type II-A was the most common, containing 0–24 spacers. Only 10% spacers were found to be homological to known bacteriophage or plasmid sequences in databases. Two Enterococcus faecium strains and a Lactococcus lactis strain carried antibiotic resistance genes. Genes encoding for ornithine decarboxylase were detected in four strains and genes encoding for agmatine deiminase were detected in four strains. Lactobacillus paraplantarum 251 L appeared to be the most interesting strain, as it contained genes encoding for two bacteriocins, a restriction–modification system, two CRISPR-Cas systems, four prophages and no genes connected with antibiotic resistance or production of biogenic amines.

Spontaneously induced prophages are abundant in a naturally evolved bacterial starter culture and deliver competitive advantage to the host

BackgroundIn complex microbial ecosystems such as the marine environment, the gastrointestinal tract, but also in mixed culture fermentations, bacteriophages are frequently found to be a part of the microbial community. Moreover, prophages or prophage-like elements are frequently identified in sequenced bacterial genomes. The mixed undefined starter cultures represent an ecosystem which is shaped by long term evolution under relatively defined environmental conditions and provides an interesting model to study co-evolution of phages and their hosts as well as the impact of diversity on microbial community stability.ResultsIn the present study we investigated the presence, identity and behaviour of prophages in lactococci being part of a complex cheese starter culture. Genome analysis of representative strains of the 7 genetic lineages of Lactococcus lactis constituting the culture indicated the presence of prophages in all strains. Exposure of potential lysogens to mitomycin C confirmed the release of ~ 1010·ml− 1 phage particles from all tested strains. Furthermore, phages were also released in substantial amounts due to spontaneous induction: more than 108·ml− 1 phage particles were present in cultures under non-inducing conditions. This observation suggests continuous release of phage particles by the lactococci. The released bacteriophages exhibited an unusual morphology. For most strains tested, tailless icosahedral phage heads were found. The competitive advantage of lysogens compared to their cured derivatives and their high abundance in the culture suggests that the released tailless bacteriophages play an important role in the ecosystem.ConclusionsThe results of this study indicate that chromosomal genetic elements are active participants in the stable complex microbial community of the starter culture. We show that prophages are abundant in such a community, are produced continuously in large amounts and, despite the huge metabolic burden imposed on the cells by phage particle production, provide a selective advantage to the host.

Exopolysaccharide producing Lactobacillus plantarum SKT109 as adjunct culture in Cheddar cheese production

In vitro antioxidant activity of an exopolysaccharide from Lactobacillus plantarum SKT109 (L-EPS) and potential application of this strain for Cheddar cheese production were evaluated. Cheddar cheese was manufactured using a cheese starter culture consisting of lactococci combined with L. plantarum SKT109 as an adjunct culture. The chemical composition, microbial changes, proteolysis, textural profiles and volatile compounds of the cheeses were studied during 12 weeks of ripening. The experimental cheese made with L. plantarum SKT109 as an adjunct had greater moisture contents than those of the control cheese made with starter alone. L. plantarum SKT109 survived well in Cheddar cheese, with counts of 7.89 log cfu/g at the end of the ripening period. Inclusion of the adjunct strain had no detectable negative effects on cheeses in terms of proteolysis and texture. L. plantarum SKT109 improved the Cheddar cheese flavor profile by increasing the levels of low threshold value compounds including ethyl acetate, 3-methyl-1-butanol, 3-hydroxy-2-butanone and butanone. L-EPS had the strongest scavenging activity toward hydroxyl (68.52%) radicals at a dose of 5 mg/mL. These results indicated that EPS-producing L. plantarum SKT109 or its EPS may be promising ingredients, with favorable effects on cheese texture and flavor as well as potential health benefits.

Characteristics of Kwark Cheese Supplemented with Bifidobacterium longum KACC 91563.

The effect of addition of the probiotic Bifidobacterium longum KACC 91563 on the chemical and sensory properties of Kwark cheese produced using CHN-11 as a cheese starter were investigated. The addition of B. longum KACC 91563 to Kwark cheese did not change the composition or pH value of the cheese, compared with control. B. longum KACC 91563 survived at a level of 7.58 Log CFU/g and did not have any negative effect on survival of the cheese starter. A sensory panel commented that the addition of B. longum KACC 91563 made Kwark cheese more desirable to consumers, and that the probiotic supplementation had no effect on perceived taste. Thus, B. longum KACC 91563 can be used for inclusion of probiotic bacteria in cheese.

Uji Kadar Lemak Keju Cheddar Dengan Variasi Bahan Baku (Sapi, Kambing) Serta Variasi Jenis Starter (Streptococcus lactis, Rhizophus oryzae).

Cheese is dairy product. Microorganism are most widely used in starter, especially the cheese starter is a group of lactic acid bacteria (LAB), which is capable of producing acid Bacteri acid lactit (BAL) often used as starter in cheesemaking. This purpose of this study is variation of milk and starter in to improve the quality.In this research the ceddar making was conducted the varation milk and starter. cheddar cheese produced in this study analyzed the value of curd,water, and fat contained with Anava then followed by DMRT 5% level of significance. The best quality in the cheddar cheese at K2B2 , the randemen value 21.66, water value 38.66 and fat value 34.51.

Characterization of volatile compounds produced by Lactobacillus helveticus strains in a hard cheese model.

Starter cultures of Lactobacillus helveticus used in hard cooked cheeses play an important role in flavor development. In this work, we studied the capacity of three strains of L. helveticus, two autochthonous (Lh138 and Lh209) and one commercial (LhB02), to grow and to produce volatile compounds in a hard cheese extract. Bacterial counts, pH, profiles of organic acids, carbohydrates, and volatile compounds were analyzed during incubation of extracts for 14 days at 37 ℃. Lactobacilli populations were maintained at 106 CFU ml-1 for Lh138, while decreases of approx. 2 log orders were found for LhB02 and Lh209. Both Lh209 and LhB02 slightly increased the acetic acid content whereas mild increase in lactic acid was produced by Lh138. The patterns of volatiles were dependent on the strain which reflect their distinct enzymatic machineries: LhB02 and Lh209 produced a greater diversity of compounds, while Lh138 was the least producer strain. Extracts inoculated with LhB02 and Lh 209 were characterized by ketones, esters, alcohols, aldehydes, and acids, whereas in the extracts with Lh138 the main compounds belonged to aromatic, aldehydes, and ketones groups. Therefore, Lh209 and LhB02 could represent the best cheese starters to improve and intensify the flavor, and even a starter composed by combinations of LhB02 or Lh209 with Lh138 could also be a strategy to diversify cheese flavor.

Production of benzoic acid as a natural compound in fermented skim milk using commercial cheese starter

In this study, we investigated the production of natural benzoic acid (BA) in skim milk fermentation by 5 kinds of commercial cheese starters. Five kinds of starter were inoculated into 10% reconstituted skim milk, and then the culture was incubated at 2-h intervals for 10 h at 30, 35, and 40°C. In fermentation by MW 046 N+LH 13, the starter for making raclette, BA was highly detected after 8 h at 30 and 35°C. In fermentation by LH 13, the starter for making berg, BA steadily increased and was highly detected at 40°C. In fermentation by TCC-3+TCC-4, the starter for making Caciocavallo and mozzarella, BA was detected after 2 h at 40°C. Also, BA was detected after 4 and 8 h at 35 and 30°C, respectively. In fermentation by Flora-Danica, the starter for making Gouda, BA was increased until 6 h and decreased after 6 h at all temperatures. Among the 5 kinds of fermentation, the level of BA was the highest in fermentation by Flora-Danica at 6 h at 35°C, at 14.55 mg/kg.

Contribution of the novel sulfur-producing adjunct Lactobacillus nodensis to flavor development in Gouda cheese

We assessed the efficacy of Lactobacillus nodensis CSK964 as an adjunct culture in Gouda cheese under various industrial conditions. We set up 4 different systems: a direct vat inoculum with and without adjunct using the calf rennet Kalase, and an undefined bulk starter culture with and without adjunct using the microbial rennet Milase (both rennets from CSK Food Enrichment, Ede, the Netherlands). During ripening, we subjected the cheeses to the following analyses: viability of starter and adjunct cells, composition, proteolysis, and flavor development by detection of sulfur compounds and descriptive sensory analysis. In general, the presence of Lb. nodensis increased secondary proteolysis and influenced cheese flavor, particularly in relation to volatile sulfur compounds; hydrogen sulfide and methanethiol were present in higher abundances in cheeses containing Lb. nodensis. The primary starter also influenced the range of volatile sulfur compounds produced. Methanethiol and dimethyl disulfide were more abundant in the nisin-producing direct vat inoculum cheese with adjunct; hydrogen sulfide was more prevalent when bulk starter culture was used with Lb. nodensis. Sensory analysis revealed that the direct vat inoculum cheese with adjunct scored significantly better in terms of smell and taste than the direct vat inoculum cheese without adjunct and lacked the dominant sulfur flavors of the bulk starter cheese with adjunct. Subsequent analysis using lead acetate paper and modified motility broth as indicators of hydrogen sulfide production confirmed that Lb. nodensis produced hydrogen sulfide in broth and in the cheese matrix. This study suggests that the inclusion of Lb. nodensis as an adjunct culture can significantly alter the flavor profile of the final cheese. However, the selection of a suitable primary starter is imperative to ensure a desirable product.

Short communication: Change of naturally occurring benzoic acid during skim milk fermentation by commercial cheese starters.

This study sought to investigate the change of naturally occurring benzoic acid (BA) during skim milk fermentation by 4 kinds of commercial cheese starters used in domestic cheese. The culture was incubated at 3-h intervals for 24h at 30, 35, and 40°C. The BA content during fermentation by Streptococcus thermophilus STB-01 was detected after 12h at all temperatures, sharply increasing at 30°C. In Lactobacillus paracasei LC431, BA was detected after 9h at all temperatures, sharply increasing until 18h and decreasing after 18h at 30 and 35°C. In the case of R707 (consisting of Lactococcus lactis ssp. lactis and Lactococcus lactis ssp. cremoris), BA increased from 6h to 15h and decreased after 15h at 40°C. The BA during STB-01 and CHN-11 (1:1; mixture of S. thermophilus, Lc. lactis ssp. lactis, Lc. lactis ssp. cremoris, Lc. lactis ssp. diacetylactis, Leuconostoc mesenteroides ssp. cremoris) fermentation was detected after 3h at 35 and 40°C, sharply increasing up to 12h and decreasing after 15h at 35°C, and after 6h, increasing up to 9h at 30°C. After 3h, it steadily decreased at 40°C. The highest amount of BA was found during the fermentation by R707 at 30°C; 15h with 12.46mg/kg.

Impact ofSpirulina platensison Physicochemical Properties and Viability ofLactobacillus acidophilusof Probiotic UF Feta Cheese

The impact of addition of powdered Spirulina platensis on the physicochemical properties and microbiological of cheese containing Lactobacillus acidophilus and Mentha longifolia L. was studied during the refrigerated storage. Cheese starter cultures (1% w/v) and L. acidophilus (2% w/v) was inoculated to the milk together with M. longifolia L. (0.5 and 1% w/v) and S. platensis (0, 0.3, 0.5 and 0.8% w/v). Produced cheeses were stored at 4C for 45 days and microbial, textural and nutritional aspects were analyzed. Practical Applications This study may have practical applications in the production of enriched cheese feta as a fermented products which needs to elongation of shelf life and enrichment (especially in critical condition). The result showed that adding the Spirulina platensis biomass significantly increased (P < 0.05) the amount of iron, protein and hardness of probiotic feta cheese during the refrigerated storage at 4C. Also according to the result of the viability of probiotic bacteria, the positive effects of S. platensis were observed on the survival of Lactobacillus acidophilus during storage of cheeses. Lightness and redness decreased by increasing content of Spirulina, in contrast, b value (yellowness) increased.

High-throughput DNA sequencing to survey bacterial histidine and tyrosine decarboxylases in raw milk cheeses.

BackgroundThe aim of this study was to employ high-throughput DNA sequencing to assess the incidence of bacteria with biogenic amine (BA; histamine and tyramine) producing potential from among 10 different cheeses varieties. To facilitate this, a diagnostic approach using degenerate PCR primer pairs that were previously designed to amplify segments of the histidine (hdc) and tyrosine (tdc) decarboxylase gene clusters were employed. In contrast to previous studies in which the decarboxylase genes of specific isolates were studied, in this instance amplifications were performed using total metagenomic DNA extracts.ResultsAmplicons were initially cloned to facilitate Sanger sequencing of individual gene fragments to ensure that a variety of hdc and tdc genes were present. Once this was established, high throughput DNA sequencing of these amplicons was performed to provide a more in-depth analysis of the histamine- and tyramine-producing bacteria present in the cheeses. High-throughput sequencing resulted in generation of a total of 1,563,764 sequencing reads and revealed that Lactobacillus curvatus, Enterococcus faecium and E. faecalis were the dominant species with tyramine producing potential, while Lb. buchneri was found to be the dominant species harbouring histaminogenic potential. Commonly used cheese starter bacteria, including Streptococcus thermophilus and Lb. delbreueckii, were also identified as having biogenic amine producing potential in the cheese studied. Molecular analysis of bacterial communities was then further complemented with HPLC quantification of histamine and tyramine in the sampled cheeses.ConclusionsIn this study, high-throughput DNA sequencing successfully identified populations capable of amine production in a variety of cheeses. This approach also gave an insight into the broader hdc and tdc complement within the various cheeses. This approach can be used to detect amine producing communities not only in food matrices but also in the production environment itself.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-015-0596-0) contains supplementary material, which is available to authorized users.

The diversity and evolution of microbiota in traditional Turkish Divle Cave cheese during ripening

The microbial diversity of traditional Turkish Divle Cave cheese was evaluated in three independent batches. Using molecular techniques, twenty three bacterial species were identified in the interior and outer part of the cheese on days 60 and 120. Bacilli and Gammaproteobacteria classes were predominant during early stages of ripening and Actinobacteria at later stages. Nineteen species of filamentous fungi and five yeast species were identified and the most frequently isolated species were Penicillium polonicum, Penicillium biforme, Penicillium roqueforti, Penicillium chrysogenum and Debaryomyces hansenii. The microbiota displayed similar communities among batches, but high diversity in different parts of the cheese during ripening. Novel cheese starter cultures could be developed after the technological characterisation of the isolated strains.

Inhibition of bacterial growth in sweet cheese whey by carbon dioxide as determined by culture-independent community profiling

Whey is a valuable co-product from cheese making that serves as a raw material for a wide range of products. Its rich nutritional content lends itself to rapid spoilage, thus it typically needs to be pasteurised and refrigerated promptly. Despite the extensive literature on milk spoilage bacteria, little is known about the spoilage bacteria of whey. The utility of carbon dioxide (CO2) to extend the shelf-life of raw milk and cottage cheese has been well established, but its application in whey preservation has not yet been explored. This study aims to characterise the microbial populations of fresh and spoiled sweet whey by culture-independent community profiling using 454 pyrosequencing of 16S rRNA gene amplicons and to determine whether carbonation is effective in inhibiting bacterial growth in sweet whey. The microbiota of raw Cheddar and Mozzarella whey was dominated by cheese starter bacteria. After pasteurisation, two out of the three samples studied became dominated by diverse environmental bacteria from various phyla, with Proteobacteria being the most dominant. Diverse microbial profiles were maintained until spoilage occurred, when the entire population was dominated by just one or two genera. Whey spoilage bacteria were found to be similar to those of milk. Pasteurised Cheddar and Mozzarella whey was spoiled by Bacillus sp. or Pseudomonas sp., and raw Mozzarella whey was spoiled by Pseudomonas sp., Serratia sp., and other members of the Enterobacteriaceae family. CO2 was effective in inhibiting bacterial growth of pasteurised Cheddar and Mozzarella whey stored at 15°C and raw Mozzarella whey stored at 4°C. The spoilage bacteria of the carbonated samples were similar to those of the non-carbonated controls.

Single-strain starter experimental cheese reveals anti-inflammatory effect of Propionibacterium freudenreichii CIRM BIA 129 in TNBS-colitis model

Propionibacterium freudenreichii is a beneficial bacterium consumed as a cheese starter and as a probiotic. It displays promising immunomodulatory properties and selected strains exert an anti-inflammatory effect via key surface proteins. Cheese constitutes an important source of bacteria, which can have beneficial effects, depending on the species or strain. We developed here a single-strain experimental pressed cheese, exclusively fermented by P. freudenreichii, in order to investigate its health effects, which is not possible in a complex cheese ecosystem. Key immunomodulatory surface proteins were expressed within the cheese matrix. Consumption of this axenic cheese protected mice from acute colitis induced by TNBS (trinitrobenzenesulphonic acid). Induction of local and systemic inflammatory markers, as well as colon epithelial oxidative stress markers, was prevented. This work confirms the probiotic potential of P. freudenreichii and provides a new functional fermented product for preclinical and clinical studies aimed at prevention or treatment of IBD.

Emmental Cheese Environment Enhances Propionibacterium freudenreichii Stress Tolerance.

Dairy propionibacteria are actinomycetales found in various fermented food products. The main species, Propionibacterium freudenreichii, is generally recognized as safe and used both as probiotic and as cheese starter. Its probiotic efficacy tightly depends on its tolerance towards digestive stresses, which can be largely modulated by the ingested delivery vehicle. Indeed, tolerance of this bacterium is enhanced when it is consumed within a fermented dairy product, compared to a dried probiotic preparation. We investigated both stress tolerance and protein neosynthesis upon growth in i) chemically defined or ii) aqueous phase of Emmental cheeses. Although the same final population level was reached in both media, a slower growth and an enhanced survival of CIRM BIA 1 strain of P. freudenreichii subsp. shermanii was observed in Emmental juice, compared to chemically defined medium. This was accompanied by differences in substrates used and products released as well as overexpression of various early stress adaptation proteins in Emmental juice, compared to chemically defined medium, implied in protein folding, in aspartate catabolism, in biosynthesis of valine, leucine and isoleucine, in pyruvate metabolism in citrate cycle, in the propionate metabolism, as well as in oxidoreductases. All these changes led to a higher digestive stress tolerance after growth in Emmental juice. Mechanisms of stress adaptation were induced in this environment, in accordance with enhanced survival. This opens perspectives for the use of hard and semi-hard cheeses as delivery vehicle for probiotics with enhanced efficacy.

Melon aroma-producing yeast isolated from coastal marine sediment in Maizuru Bay, Japan

Researches on marine fungi and fungi isolated from marine environments are not active compared with those on terrestrial fungi. The aim of this study was isolation of novel and industrially applicable fungi derived from marine environments. In this study, 16 fungus-like strains, MS1–MS16, were isolated from coastal marine sediment in Maizuru Bay, Japan, under aerobic culture conditions. Phylogenetic analysis of 18S rRNA gene sequences indicated that 10 isolates belonged to Ascomycota, five isolates belonged to Sordariomycetes, two were Dothideomycetes, and three were Saccharomycetes. Liquid and agar potato dextrose cultures of strains MS1 and MS2 isolated from the coastal sediment released a melon-like aroma. Gas chromatography analysis suggested that strains MS1 and MS2 produce four major chemicals associated with a melon aroma, cis-3-hexen-1-ol, cis-6-nonenal, 3,6-nonadien-1-ol, and trans,cis-2,6-nonadienal. The sequence analyses of the 26S rRNA domains 1/2 (D1/D2) and internal transcribed spacer regions indicated that strains MS1 and MS2 were phylogenetically identified as Geotrichum candidum, a well-known yeast used as a cheese starter. These results suggest the future isolations of novel and functional fungi from marine environments.

Application of 16S-ARDRA and RFLP-PFGE for improved genotypic characterisation of dairy propionibacteria and combination with characteristic phenotypes

Propionic acid bacteria (PAB) are important for the manufacture of Swiss-type cheeses since they are involved in the formation of eyes and are responsible for the characteristic flavour and aroma of these cheeses. Propionibacterium freudenreichii is most commonly encountered in Swiss-type cheeses and is the sole Propionibacterium species used as a cheese starter. Aspartase activity determined in PAB is strain-dependent and it is important to find a usable way to identify the specific strains involved in cheese making. In this work, 16S-amplified ribosomal DNA restriction analysis and pulsed-field gel electrophoresis (PFGE) were used to differentiate 107 PAB strains. Consequently, the presence and persistence of inoculated Propionibacterium starter strains can be monitored during cheese ripening. The studied metabolic features of lactose utilisation, nitrate reductase activity and aspartase activity could not be directly associated with particular genotypes, but principal component analysis (PCA) suggested a linkage between related PFGE patterns and aspartase activity.