Solange Buchin

Traditional cheeses: Rich and diverse microbiota with associated benefits

The risks and benefits of traditional cheeses, mainly raw milk cheeses, are rarely set out objectively, whence the recurrent confused debate over their pros and cons. This review starts by emphasizing the particularities of the microbiota in traditional cheeses. It then describes the sensory, hygiene, and possible health benefits associated with traditional cheeses. The microbial diversity underlying the benefits of raw milk cheese depends on both the milk microbiota and on traditional practices, including inoculation practices. Traditional know-how from farming to cheese processing helps to maintain both the richness of the microbiota in individual cheeses and the diversity between cheeses throughout processing. All in all more than 400 species of lactic acid bacteria, Gram and catalase-positive bacteria, Gram-negative bacteria, yeasts and moulds have been detected in raw milk. This biodiversity decreases in cheese cores, where a small number of lactic acid bacteria species are numerically dominant, but persists on the cheese surfaces, which harbour numerous species of bacteria, yeasts and moulds. Diversity between cheeses is due particularly to wide variations in the dynamics of the same species in different cheeses. Flavour is more intense and rich in raw milk cheeses than in processed ones. This is mainly because an abundant native microbiota can express in raw milk cheeses, which is not the case in cheeses made from pasteurized or microfiltered milk. Compared to commercial strains, indigenous lactic acid bacteria isolated from milk/cheese, and surface bacteria and yeasts isolated from traditional brines, were associated with more complex volatile profiles and higher scores for some sensorial attributes. The ability of traditional cheeses to combat pathogens is related more to native antipathogenic strains or microbial consortia than to natural non-microbial inhibitor(s) from milk. Quite different native microbiota can protect against Listeria monocytogenes in cheeses (in both core and surface) and on the wooden surfaces of traditional equipment. The inhibition seems to be associated with their qualitative and quantitative composition rather than with their degree of diversity. The inhibitory mechanisms are not well elucidated. Both cross-sectional and cohort studies have evidenced a strong association of raw-milk consumption with protection against allergic/atopic diseases; further studies are needed to determine whether such association extends to traditional raw-milk cheese consumption. In the future, the use of meta-omics methods should help to decipher how traditional cheese ecosystems form and function, opening the way to new methods of risk-benefit management from farm to ripened cheese.

Ripening and quality of Swiss-type cheese made from raw, pasteurized or microfiltered milk

Abstract Experimental mini-cheeses were made from raw (Ra), microfiltered (MF), pasteurized (Pa) (72 °C, 30s) or pasteurized mixed with microfiltration retentate (PR) milk to study the influence of the indigenous microflora and pasteurization on the quality of Swiss-type cheese. To estimate biochemical transformations during cheese ripening, several methods were used: nitrogen fractionation (water-soluble fraction and phosphotungstic acid (PTA)-soluble fraction), urea-polyacrylamide gel electrophoresis of caseins, reverse phase liquid chromatography of the water-soluble fraction, lactate and volatile fatty acids. Microbial populations were also enumerated. At the end of ripening, in comparison with MF and Pa milk cheeses, Ra and PR milk cheeses exhibited higher overall aroma intensity and pungency, characteristics which correlated with higher populations of facultatively heterofermentative lactobacilli (108 cfug−1), propionibacteria (108 cfug−1), and enterococci (106cfug−1). These cheeses had high levels of PTA-soluble N and acetic, propionic and isovaleric acids. MF and Pa milk cheeses, although somewhat different from one another, were very different from the two other types of cheese. Pa milk cheese had a lower pH than MF milk cheese, and contained a higher proportion of γ-caseins due to the activation of plasmin. Moreover, Pa milk cheese was more acidic, but demonstrated a higher overall aroma intensity. The addition of raw milk flora (retentate) to Pa milk restored almost all the biochemical and sensory characteristics of Ra milk cheese measured in this study.

Effect of autochthonous lactic acid bacteria starters on health-promoting and sensory properties of tomato juices.

Strains of Lactobacillus plantarum, Weissella cibaria/confusa, Lactobacillus brevis, Pediococcus pentosaceous, Lactobacillus sp. and Enterococcus faecium/faecalis were identified from raw tomatoes by Biolog System, partial 16S rRNA gene sequence and subjected to typing by Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis. Ten autochthonous strains were singly used to ferment tomato juice (TJ) via a protocol which included fermentation at 25 degrees C for 17 h and further storage at 4 degrees C for 40 days. Unstarted TJ and TJ fermented with an allochthonous strain of L. plantarum were used as the controls. All autochthonous strains grew well in TJ reaching cell densities ca. 10,000 and 10 times higher than unstarted TJ and TJ fermented with the allochthonous strain. Viscosity of TJs fermented with autochthonous strains was generally the highest, especially when started with W. cibaria/confusa which synthesized exo-polysaccharides. Overall, unstarted TJ and TJ fermented with the allochthonous strain showed marked decreases of ascorbic acid (ASC), glutathione (GSH) and total antioxidant activity (TTA) during storage. On the contrary, several TJs fermented with autochthonous strains, especially with L. plantarum POM1 and POM 35, maintained elevated values of ASC, GSH and TAA. The variation of color indexes mirrored the above behavior. TJs fermented with the above two autochthonous strains were compared to controls based on volatile components through Purge and Trap or Solid Phase Microextraction Gas Chromatography-Mass Spectrometry (PT or SPME-GC/MS) analysis. As shown by Principal Component Analysis a large number of volatiles belonging to various chemical classes markedly differentiated TJs fermented with autochthonous strains with respect to controls.

Metabolic responses of Lactobacillus plantarum strains during fermentation and storage of vegetable and fruit juices.

ABSTRACT Strains of Lactobacillus plantarum were grown and stored in cherry (ChJ), pineapple (PJ), carrot (CJ), and tomato (TJ) juices to mimic the chemical composition of the respective matrices. Wheat flour hydrolysate (WFH), whey milk (W), and MRS broth were also used as representatives of other ecosystems. The growth rates and cell densities of L. plantarum strains during fermentation (24 h at 30°C) and storage (21 days at 4°C) differed only in part, being mainly influenced by the matrix. ChJ and PJ were the most stressful juices for growth and survival. Overall, the growth in juices was negatively correlated with the initial concentration of malic acid and carbohydrates. The consumption of malic acid was noticeable for all juices, but mainly during fermentation and storage of ChJ. Decreases of branched-chain amino acids (BCAA)—with the concomitant increase of their respective branched alcohols—and His and increases of Glu and gamma-aminobutyric acid (GABA) were the main traits of the catabolism of free amino acids (FAA), which were mainly evident under less acidic conditions (CJ and TJ). The increase of Tyr was found only during storage of ChJ. Some aldehydes (e.g., 3-methyl-butanal) were reduced to the corresponding alcohols (e.g., 3-methyl-1-butanol). After both fermentation and storage, acetic acid increased in all fermented juices, which implied the activation of the acetate kinase route. Diacetyl was the ketone found at the highest level, and butyric acid increased in almost all fermented juices. Data were processed through multidimensional statistical analyses. Except for CJ, the juices (mainly ChJ) seemed to induce specific metabolic traits, which differed in part among the strains. This study provided more in-depth knowledge on the metabolic mechanisms of growth and maintenance of L. plantarum in vegetable and fruit habitats, which also provided helpful information to select the most suitable starters for fermentation of targeted matrices.

In vivo sodium release and saltiness perception in solid lipoprotein matrices. 1. Effect of composition and texture

Reducing the sodium content in foods is complex because of their multidimensional sensory characteristics and the multifunctionality of sodium chloride. The aim of this study was to elucidate how food composition may influence in-mouth sodium release and saltiness perception. Lipoprotein matrices (LPM) were produced using milk constituents and characterized by means of rheological measurements, texture, and taste sensory profiles. Texture and taste perceptions were affected differently by variations in the salt level, dry matter, and fat contents. Composition and textural changes also modified temporal sodium release and saltiness perception recorded in five subjects, but the effects varied as a function of the salt content. The water content mainly appeared to influence the amount of sodium released, whereas saltiness perception was mainly related to fat content. Elasticity, coating, and granularity were found to be correlated with temporal sodium release and/or saltiness parameters.

Diversity of the lactic acid bacterium and yeast microbiota in the switch from firm- to liquid-sourdough fermentation.

ABSTRACT Four traditional type I sourdoughs were comparatively propagated (28 days) under firm (dough yield, 160) and liquid (dough yield, 280) conditions to mimic the alternative technology options frequently used for making baked goods. After 28 days of propagation, liquid sourdoughs had the lowest pH and total titratable acidity (TTA), the lowest concentrations of lactic and acetic acids and free amino acids, and the most stable density of presumptive lactic acid bacteria. The cell density of yeasts was the highest in liquid sourdoughs. Liquid sourdoughs showed simplified microbial diversity and harbored a low number of strains, which were persistent. Lactobacillus plantarum dominated firm sourdoughs over time. Leuconostoc lactis and Lactobacillus brevis dominated only some firm sourdoughs, and Lactobacillus sanfranciscensis persisted for some time only in some firm sourdoughs. Leuconostoc citreum persisted in all firm and liquid sourdoughs, and it was the only species detected in liquid sourdoughs at all times; it was flanked by Leuconostoc mesenteroides in some sourdoughs. Saccharomyces cerevisiae, Candida humilis, Saccharomyces servazzii, Saccharomyces bayanus-Kazachstania sp., and Torulaspora delbrueckii were variously identified in firm and liquid sourdoughs. A total of 197 volatile components were identified through purge and trap–/solid-phase microextraction–gas chromatography-mass spectrometry (PT–/SPME–GC-MS). Aldehydes, several alcohols, and some esters were at the highest levels in liquid sourdoughs. Firm sourdoughs mainly contained ethyl acetate, acetic acid, some sulfur compounds, and terpenes. The use of liquid fermentation would change the main microbial and biochemical features of traditional baked goods, which have been manufactured under firm conditions for a long time.

Simplification of a complex microbial antilisterial consortium to evaluate the contribution of its flora in uncooked pressed cheese

A complex microbial consortium derived from raw milk and composed of populations classified in 4 groups (lactic acid bacteria (A), Gram positive catalase positive bacteria (B), Gram negative bacteria (C) and yeasts (D)) can contribute to the inhibition of Listeria monocytogenes in the core of an uncooked pressed cheese. To identify what groups may be involved in the inhibition, the consortium was simplified by successively omitting one group at a time. Pasteurized milk was inoculated with these more or less complex consortia and their effects on L. monocytogenes count, pH, acids and volatile compounds in the core of uncooked pressed cheese were evaluated. The growth of L. monocytogenes was the highest in cheeses prepared with pasteurized milk and only St. thermophilus. Inhibition in other cheeses was expressed by comparison with growth in these ones. All the consortia containing both lactic acid bacteria (group A) and Gram positive catalase positive bacteria (group B)--ABCD, ABD, ABC, AB--were more inhibitory than those containing lactic acid bacteria on its own (A) or associated only with yeasts (AD) or/and Gram negative (ADC). Consortia without lactic acid bacteria were weakly inhibitory or had no effect. Gram positive catalase positive bacteria alone were not inhibitory although most of the species became established in the cheeses. The Lactobacillus population (Lb. casei, Lb. plantarum, Lb. curvatus and Lb. farciminis) was predominant in cheeses (9 log CFU/g) with a higher count than Leuconostoc (7 log CFU/g) and Enterococcus (7 log CFU/g). Lactobacillus counts were negatively correlated with those of L. monocytogenes (r=-0.84 at 18 days) and with the level of D-lactic acid. There was no correlation between L. monocytogenes and Leuconostoc or Enterococcus counts. Complex consortium ABCD and AB not only had a stronger inhibitory power in cheeses than consortium AD, they were also associated with the highest levels of L-lactic and acetic acids. All cheeses inoculated with lactic acid bacteria differed from those without by higher levels of ethyl formiate, pentane and alcohols (2-butanol, 2-pentanol), and lower levels of ketones (2-hexanone, 2,3-butanedione) and aldehydes (2-methyl-butanal). Levels of 2-methyl-butanal, 2-butanol and 2-pentanol were higher in ABCD and AB cheeses than in AD cheeses. Beside their contribution to the inhibition, their effect on cheese flavour must be evaluated.

Use of microparticulated whey protein concentrate, exopolysaccharide-producing Streptococcus thermophilus, and adjunct cultures for making low-fat Italian Caciotta-type cheese.

Low-fat Caciotta-type cheeses were manufactured with partially skim milk (fat content of ~0.3%) alone (LFC); with the supplementation of 0.5% (wt/vol) microparticulated whey protein concentrate (MWPC) (LFC-MWPC); with MWPC and exopolysaccharides (EPS)-producing Streptococcus thermophilus ST446 (LFC-MWPC-EPS); and with MWPC, EPS-producing strain ST446, and Lactobacillus plantarum LP and Lactobacillus rhamnosus LRA as adjunct cultures (LFC-MWPC-EPS-A). The non-EPS-producing isogenic variant Streptococcus thermophilus ST042 was used for making full-fat Caciotta-type cheese (FFC), LFC, and LFC-MWPC. Cheeses were characterized based on compositional, microbiological, biochemical, texture, volatile components (purge and trap, and solid-phase microextraction coupled with gas chromatography-mass spectrometry), and sensory analyses. Compared with FFC and LFC (51.6 ± 0.7 to 53.0 ± 0.9%), the other cheese variants retained higher levels of moisture (60.5 ± 1.1 to 67.5 ± 0.5%). The MWPC mainly contributed to moisture retention. Overall, all LFC had approximately one-fourth (22.6 ± 0.8%) of the fat of FFC. Hardness of cheeses slightly varied over 7d of ripening. Microbial EPS positively affected cheese texture, and the texture of LFC without MWPC or microbial EPS was excessively firm. Free amino acids were at the highest levels in LFC treatments (2,705.8 ± 122 to 3,070.4 ± 123 mg/kg) due to the addition of MWPC and the peptidase activity of adjunct cultures. Aldehydes, alcohols, ketones, sulfur compounds, and short- to medium-chain carboxylic acids differentiated LFC variants and FFC. The sensory attributes pleasant to taste, intensity of flavor, overall acceptability, and pleasant to chew variously described LFC-MWPC-EPS and LFC-MWPC-EPS-A. Based on the technology options used, low-fat Caciotta-type cheese (especially ripened for 14 d) has promising features to be further exploited as a suitable alternative to the full-fat variant.

Distribution and mobility of phosphates and sodium ions in cheese by solid-state 31P and double-quantum filtered 23Na NMR spectroscopy

The feasibility of solid‐state magic angle spinning (MAS) 31P nuclear magnetic resonance (NMR) spectroscopy and 23Na NMR spectroscopy to investigate both phosphates and Na+ ions distribution in semi‐hard cheeses in a non‐destructive way was studied. Two semi‐hard cheeses of known composition were made with two different salt contents. 31P Single‐pulse excitation and cross‐polarization MAS experiments allowed, for the first time, the identification and quantification of soluble and insoluble phosphates in the cheeses. The presence of a relatively ‘mobile’ fraction of colloidal phosphates was evidenced. The detection by 23Na single‐quantum NMR experiments of all the sodium ions in the cheeses was validated. The presence of a fraction of ‘bound’ sodium ions was evidenced by 23Na double‐quantum filtered NMR experiments. We demonstrated that NMR is a suitable tool to investigate both phosphates and Na+ ions distributions in cheeses. The impact of the sodium content on the various phosphorus forms distribution was discussed and results demonstrated that NMR would be an important tool for the cheese industry for the processes controls. Copyright

Spatial Distribution of the Metabolically Active Microbiota within Italian PDO Ewes' Milk Cheeses.

Italian PDO (Protected Designation of Origin) Fiore Sardo (FS), Pecorino Siciliano (PS) and Pecorino Toscano (PT) ewes’ milk cheeses were chosen as hard cheese model systems to investigate the spatial distribution of the metabolically active microbiota and the related effects on proteolysis and synthesis of volatile components (VOC). Cheese slices were divided in nine sub-blocks, each one separately subjected to analysis and compared to whole cheese slice (control). Gradients for moisture, and concentrations of salt, fat and protein distinguished sub-blocks, while the cell density of the main microbial groups did not differ. Secondary proteolysis differed between sub-blocks of each cheese, especially when the number and area of hydrophilic and hydrophobic peptide peaks were assessed. The concentration of free amino acids (FAA) agreed with these data. As determined through Purge and Trap (PT) coupled with Gas Chromatography-Mass Spectrometry (PT-GC/MS), and regardless of the cheese variety, the profile with the lowest level of VOC was restricted to the region identified by the letter E defined as core. As shown through pyrosequencing of the 16S rRNA targeting RNA, the spatial distribution of the metabolically active microbiota agreed with the VOC distribution. Differences were highlighted between core and the rest of the cheese. Top and bottom under rind sub-blocks of all three cheeses harbored the widest biodiversity. The cheese sub-block analysis revealed the presence of a microbiota statistically correlated with secondary proteolysis events and/or synthesis of VOC.