Genotypic and technological characterization of Leuconostoc isolates to be used as adjunct starters in Manchego cheese manufacture

Twenty-seven Leuconostoc (Ln.) isolates from Manchego cheese were characterized by phenotypic and genotypic methods, and their technological abilities studied in order to test their potential use as dairy starter components. While phenotypic diversity was evaluated by studying the biochemical characteristics of technological interest (i.e. acidifying and aminopeptidase activities), genotypic diversity was evidenced by using Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR). Additional technological abilities such as lipolytic, proteolytic and autolytic activities, salt and pH tolerance and production of dextran, flavour compounds and biogenic amines, were investigated. The marked differences among strains reflected the existing biodiversity in naturally fermented products. After statistically evaluating their performance, strains C0W2, belonging to Ln. lactis, and C16W5 and N2W5, belonging to Ln. mesenteroides subsp. dextranicum, revealed the best properties to be used in mixed dairy starter cultures. This study evidences the fact that natural environments can be considered as a proper source of useful strains, for the dairy industry.     

Preliminary observations on proteolysis in Manchego cheese made with a defined-strain starter culture and adjunct starter (Lactobacillus plantarum) or a commercial starter

Different authors have demonstrated the potential of adding lactobacilli as adjunct cultures to pasteurized milk used in cheese manufacture. The aim of this work was to observe the effect of the use of a defined-strain starter culture and the addition of an adjunct culture (Lactobacillus plantarum) to cheesemilk in order to determine their effect on the ripening of Manchego cheese. Manchego cheeses were manufactured using one of the following starter culture systems: a defined starter consisting of Lactococcus lactis ssp. lactis and Leuconostoc mesenteroides ssp. dextranicum; a defined starter, as described above, and Lb. plantarum, which were isolated from a good quality Manchego cheese made from raw milk, or a commercial starter comprised of two strains of Lc. lactis. The cheeses were sampled at 15, 45, 90 and 150 d of ripening. Principal component analysis of peak heights of reversed-phase HPLC chromatograms of 70% (v/v) ethanol-insoluble and -soluble fractions distributed the samples according to the starter used in their manufacture. Quantitative differences in several peptides were evident between the three cheeses. Cheeses made with the defined-strain starters received higher scores for the flavour quality and intensity and for overall impression than the cheeses made with the commercial starter.     

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.     

Influence of exopolysaccharide‐producing cultures on the volatile profile and sensory quality of low‐fat Tulum cheese during ripening

The objective of this investigation was to compare the composition and changes in the concentration of volatiles in low‐fat and full‐fat Tulum cheeses during ripening. Tulum cheese was manufactured from low‐ or full‐fat milk using exopolysaccharide (EPS)‐producing or non‐EPS‐producing starter cultures. A total of 82 volatile compounds were identified belonging to the following chemical groups: acids (seven), esters (21), ketones (14), aldehydes (six), alcohols (14) and miscellaneous compounds (20). The relative amounts of acids, alcohols and aldehydes increased in the cheeses made with EPS‐producing cultures during 90 days of ripening. Differences were found in the volatile profile of full‐fat Tulum cheese compared with the low‐fat variant, especially after 90 days of ripening. Exopolysaccharide‐producing cultures changed the volatile profile, and the EPS‐producing cultures including Streptococcus thermophilus + Lactobacillus delbrueckii subsp. bulgaricus + Lactobacillus helveticus (LF‐EPS2) produced cheese with higher levels of methyl ketones and aldehydes than the non‐EPS cultures. In the sensory analysis, full‐fat Tulum cheeses and the cheese produced with the EPS‐producing culture containing Lb. helveticus (LF‐EPS2) were preferred by the expert panel. It was concluded that the use of EPS‐producing starter cultures in the manufacture of low‐fat Tulum cheese had the potential to improve the flavour.     

Influence of an adjunct culture of Lactobacillus on the free amino acids and volatile compounds in a Roncal-type ewe’s-milk cheese

Free amino acids and volatile compounds were analysed in a Roncal-type cheese made from pasteurized ovine milk with and without an added adjunct culture (Lactobacillus paracasei + Lactobacillus plantarum). In both batches, the total free amino acid concentration increased 11–12-fold with ripening time, the main amino acids being Leu, Glu, Lys, Phe, Val, and Ile. At the end of ripening, significant differences were recorded for Leu, Ile, Gaba, Phe, Pser, Ser, Gin, Ala, and Orn.     

Selection, application and monitoring of Lactobacillus paracasei strains as adjunct cultures in the production of Gouda-type cheeses

Raw milk cheeses have more intense flavours than cheeses made from pasteurized milk and harbour strains with potential adjunct properties. Two Lactobacillus paracasei strains, R-40926 and R-40937, were selected as potential adjunct cultures from a total of 734 isolates from good quality artisan raw milk Gouda-type cheeses on the basis of their prevalence in different cheese types and/or over several production batches, safety and technological parameters. Conventional culturing, isolation and identification and a combined PCR-DGGE approach using total cheese DNA extracts and DNA extracts obtained from culturable fractions were employed to monitor viability of the introduced adjuncts and their effect on the cheese microbiota. The control cheese made without adjuncts was dominated by members of the starter, i.e. Lactococcus lactis and Leuconostoc pseudomesenteroides. In the cheeses containing either R-40926 or R-40937, the respective adjuncts increased in number as ripening progressed indicating that both strains are well adapted to the cheese environment and can survive in a competitive environment in the presence of a commercial starter culture. Principal component analysis of cheese volatiles determined by steam distillation-extraction and gas chromatography-mass spectrometry could differentiate cheeses made with different concentrations of adjunct R-40926 from the control cheese, and these differences could be correlated to the proteolytic and lipolytic properties of this strain. Collectively, results from microbiological and metabolic analyses indicate that the screening procedure followed throughout this study was successful in delivering potential adjunct candidates to enrich or extend the flavour palette of artisan Gouda-type cheeses under more controlled conditions.     

Effect of mesophilic lactobacilli and enterococci adjunct cultures on the final characteristics of a microfiltered milk Swiss-type cheese

The effect of four associations of adjunct cultures composed of mesophilic lactobacilli and enterococci, either solely or combined, on the microbiological, biochemical and sensory characteristics of Swiss-type cheese made using microfiltered cows’ milk and supplemented with propionibacteria was studied. The global pattern of growth was similar to that generally observed in raw milk cheese and interactions between microflora were highlighted during ripening. Enterococci, which negatively affected the survival of streptococci starters, seemed to play a limited role in the formation of volatile compounds, probably due to their low levels throughout ripening. On the contrary, mesophilic lactobacilli, which affected the evolution of propionibacteria, enterococci and L. delbrueckii subsp. lactis starter counts, modified free amino acid content, production of volatile compounds and organoleptic properties of mature cheese. This population appeared to be of major importance in the formation of cheese flavor as it was positively related to numerous potential flavor compounds such as alcohols and their corresponding esters, acetaldehyde and 4-methyl-4-heptanone. The original mesophilic lactobacilli present in milk could play an important role in the sensorial diversity of raw milk Swiss-type cheeses such as Comte.     

The effect of different dietary treatments on volatile organic compounds and aromatic characteristics of buffalo Mozzarella cheese

The objective of this study was to evaluate the effect of two dietary treatments, ryegrass hay (RH) and wrapped ryegrass silage (WRS) on the volatile organic compounds (VOCs) of buffalo Mozzarella cheese. The primary data were collected from 40 female buffaloes. Twenty animals were fed RH, and other twenty received WRS. Volatile organic compounds were identified using a mass selective detector after extraction with a ‘purge and trap’ system and a gas chromatographic analysis. Our results showed differences in the aromatic profile of Mozzarella cheese samples obtained from buffaloes receiving different feed. In particular, 5 VOCs allowed the discrimination between cheese samples.     

Taste and flavor of artisan and industrial Manchego cheese as influenced by the water-soluble extract compounds

The correlation between chemical and sensory analysis of the water-soluble extract compounds with molecular weight less than 1,000 Da (WSE < 1,000 Da) from Manchego cheese at different stages of ripening, as well as the influence of the pasteurization processes, has been studied. Free amino acids, peptides and volatile compounds were more abundant in raw milk cheeses WSE < 1,000 Da. Further fractionation of the WSE < 1,000 Da was carried on by gel permeation chromatography and amino acids, peptides, ions and volatiles were determined in the fractions. The same fractions were also subjected to organoleptic evaluation. Volatile compounds were related to the aroma and small peptides and amino acids to the taste. Volatile compounds were mainly alcohols and ketones. Fresh and floral notes were detected in the 4-month-old cheeses, which evolved into more intense and complex flavors in the 8-month-old cheeses. Peptides detected in the fractions were mainly hydrophilic. Umami taste was predominant in most of the fractions and it could be related to the presence of glutamic and aspartic acids in these fractions. Salty taste was perceived with high intensity in the fractions with a high content of salts. Bitter taste was mainly detected in the last eluting fractions and it was attributed to the Tyr, Phe and Trp residues found in these fractions. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Effect of commercial adjunct lactobacilli on biochemical and sensory characteristics of Iranian white-brined cheese

Iranian white-brined cheese was made in open vats using commercial adjunct culture of Lactobacillus helveticus Lh.Bo2 in viable and freeze-shocked (FS) forms. Biochemical and sensory characteristics of the trial cheese were studied during 2 months of ripening. Assessment of primary proteolysis by SDS-PAGE and water soluble nitrogen for the trial cheese showed no or small differences throughout ripening. However, the experimental cheeses exhibited significantly greater rates of free amino group formation. Lipolysis as measured by total free fatty acid was consistently higher in cheese made with the FS adjunct culture. Expert panellists detected significant differences between the control and the experimental cheese.     

Changes of organic acids,volatile aroma compounds and sensory characteristics of Halloumi cheese kept in brine

This study examines the changes in organic acids, volatile aroma compounds and sensory characteristics of ovine Halloumi cheese kept in brine (∼10% NaCl) at 4 °C for 45 days. The highest total score of the organoleptic assessment was observed at day 1, before storage in brine. During cheese storage, a large reduction of lactose was observed, especially in the first stages of storage while, concurrently, a significantly progressive increase in lactic and acetic acids was observed. Among the volatile aroma compounds determined were alcohols, aldehydes, ketones, acids, esters, hydrocarbons, sulphur compounds, as well as a variety of other compounds in very small quantities. Ethanol and acetic acid were the dominant volatile aromatic compounds and their concentrations increased during storage. The lipolysis of Halloumi cheese during storage was not excessive. The dominant free fatty acids were palmitic, oleic and acetic.     

Effect of Penicillium roqueforti and incorporation of whey cheese on volatile profiles and sensory characteristics of mould‐ripened Civil cheese

In this study, four different types of mould‐ripened Civil cheese were manufactured. A defined (nontoxigenic) strain of a Penicillium roqueforti (SC 509) was used as secondary starter for the manufacture of mould‐ripened Civil cheese with and without addition of the whey cheese Lor; in parallel, secondary starter‐free counterparts were manufactured. A total of 83 compounds were identified. Ketones, alcohols and esters were the principal classes of volatile components. Principal component analysis of the headspace volatiles grouped cheeses by age and type. P. roqueforti inoculated cheese was clearly separated from the other cheeses at 180 days of ripening, and these cheeses were characterised with high levels of ketones (e.g., 2‐butanone, 2‐heptanone). Differences in the panel scores between the cheese samples were not significant during the first stage of ripening (up to 60 days); as ripening proceeded, these differences were become evident and P. roqueforti inoculated cheeses received higher scores than others. Addition of Lor in the manufacture of mould‐ripened Civil cheese caused lower points by the sensory panel, and the cheese inoculated with P. roqueforti and Lor‐free was the best type of mould‐ripened Civil cheese. The results showed that the use of P. roqueforti in the manufacture of mould‐ripened Civil cheese has significant impact on the volatile profiles and sensory attributes.     

Swiss-type and Swiss–Cheddar hybrid-type cheeses: effects of manufacture on sensory character and relationships between the sensory attributes and volatile compounds and gross compositional constituents

The effects of manufacturing steps, including rate of acidification in the vat, hot room step, curd wash and ripening temperature, on the sensory character of Swiss-type and Swiss–Cheddar hybrid-type cheeses were determined. In addition, relationships were determined between sensory attributes of cheeses and their gross compositional constituents and volatile compounds. Ten assessors described the sensory characteristics using 12 odour, 19 flavour, four appearance and nine texture attributes. Gross compositional constituents were determined using standard methods. Volatile compounds were isolated using a model mouth apparatus that included a mastication device. Statistical methods were used to reduce the original sensory vocabulary to four odour, eight flavour, three appearance and five texture attributes. Results of a principal component analysis on the sensory data showed clear differences in sensory character between cheese types. For example, the Swiss-type with a low rate of acid production in the vat and a hot room step had a more 'nutty' and 'sweet' flavour than the modified Swiss-type with higher acid production and no hot room step. However, the modified Swiss–Cheddar hybrid with a curd wash step, a higher acid production and no hot room step also had a 'nutty' and 'sweet' flavour. Ripening temperature also has an effect on sensory character. Relationships between sensory character and volatile compounds and/or gross compositional constituents were determined by using partial least squares regression. One odour and one flavour attribute were shown to be correlated with subsets of volatile compounds and gross compositional constituents. Four texture attributes were correlated with subsets of gross compositional constituents. The availability of information on the manufacturing process increased the interpretability of the models.     

Relationship between sensory quality and volatile compounds of elderflower (Sambucus nigra L.) extracts

Elderflower samples (Sambucus nigra L.) of 89 different genotypes were picked at optimum flowering time in Austria, England and Denmark and extracted with an aqueous solution containing sucrose, citric acid, ascorbic acid, and sodium benzoate, and investigated for the emission of volatile compounds. Volatile compounds emitted from elderflower extracts were collected by dynamic headspace technique (purge and trap) and analysed by GC–FID and GC–MS. A total of 59 volatile compounds were identified and quantified, and significant differences among the investigated elderflower extracts were observed for 58 compounds. Diluted elderflower extracts was evaluated by sensory panels trained in determination of elderflower flavour and the results from the sensory evaluation was correlated to the content of volatile compounds in the extracts using classical factor analysis including determination of eigenvalues, factor loadings and factor scores. Four principal components (PC1–PC4), which included 28 volatile compounds and sensory quality, were extracted from the factor analysis. The four PCs explained, altogether 73.7% of the data variation [PC1 (38.5%), PC2 (19.7%), PC3 (8.1%) and PC4 (7.4%)]. Important contributors to the floral and elderflower flavour of the extracts were rose oxides, hotrienol, linalool, linalool derivatives and α-terpineol, whereas the fruitiness and freshness of the extracts were mainly due to non-oxidized monoterpenes, aliphatic aldehydes and alcohols. Possibilities for selection of cultivars with specific flavours are discussed.     

Influence of filtration on volatile compounds and sensory profile of virgin olive oils

The influence of filtration through a hydrophilic cotton layer on volatile compounds, sensory characteristics and colour of two monovarietal oils was investigated in this study. Volatiles were evaluated using headspace solid-phase microextraction-gas chromatography. After the filtration of Bu?a oils only a slight increase in total alcohols was noticed. In filtered ?rna oils a significant decrease of total alcohols and slight changes in total aldehydes, total ketones and total C5 volatile compounds concentration were detected. No significant influence on the sensory scores of oils, but some slight changes in sensorial profiles were noted (slightly higher intensities of sensory characteristics apple and grass, and higher values of the lightness L^∗ in filtered samples). The results point to unequal filtration impact on different monovarietal oils and could be useful in developing targeted technologies for specific monovarietal oils quality improvement.     

The relationship between sensory quality and volatile compounds in raw juice processed from elderberries (Sambucus nigra L.)

Elderberry juice was processed by pressing of 101 samples of elderberries (Sambucus nigra L.) picked in Austria, England and Denmark at optimum maturity. Taste panels trained in determination of elderberry flavour evaluated the juices and the results from the sensory evaluation were correlated to the content of volatile compounds. The volatile compounds were isolated from elderberry juices by dynamic headspace technique and analysed by GC-FID and GC-MS. A total of 59 volatile compounds were identified, 9 of which have not previously been detected in elderberry products. A total of 53 compounds occurred in quantifiable amounts and significant differences among the investigated elderberry samples were observed for 50 compounds. The importance of the isolated volatile compounds to elderberry flavour was determined by classical factor analysis using the results from the sensory evaluation and information on the odour of the individual volatile compounds. Classical factor analysis were used including determination of eigenvalues, factor loadings and factor scores by varimax rotation of standardised variables for extraction of four non correlated factors (principal components) explaining approximately 73% of the variation in elderberry flavour. Twenty-eight volatile compounds and sensory quality were included in the four factors. Using the compounds with highest numerical values (loadings) the four factors were named: fresh-elderberry-elder flower (35.2%), fresh-fruity-sweet (17.3%), fresh-fruity (13.7%) and green-floral-fruity (6.7%). Three compounds with characteristic elderberry odour (-damascenone, nonanal, dihydroedulan) were closely correlated with the sensory quality of elderberry together with compounds with fruity, floral, green and/or miscellaneous odours.     

The effect of heat treatment and starter culture on colour intensity and sensory properties of Kulek cheese

The effect of the heat treatment on colour intensity and sensory properties of Kulek cheese made from raw milk with (RS) or without starter culture (R) and heated milk with starter culture (HS) was investigated during ripening. Colour L, a and b values of Kulek cheeses were determined for both interior and surface. The values for cheese R were 82.4 and 74.5 for L, ?5.2 and ?5.5 for a, 31.0 and 34.7 for b interior and surface respectively. The equivalents for RS cheese were 82.8 and 75.7 for L, ?4.8 and ?5.0 for a, 31.0 and 34.0 for b. The values for HS cheese were 88.9 and 88.4 for L and ?3.3 and ?3.3 for a, 22.8 and 24.2 for b. L values (lightness) were the highest in cheeses made from heated milk while b and negative a values were the highest in cheeses made from raw milk. Aroma scores of raw and heat‐treated milk cheeses made with starter were not significantly different. The panelists scored cheeses made from raw milk without starter as the best in body and texture.     

Physicochemical and textural characteristics and volatile compounds of semihard goat cheese as affected by starter cultures

Today, cheese is valued because of its high nutritional value and unique characteristics. Improving the texture and flavor of cheese by selecting suitable starter cultures is an important way to promote the development of cheese industry. The effect of starter cultures on the physicochemical and textural properties and volatile compounds during the ripening of semihard goat cheese were investigated in this work. Different starter cultures—mesophilic (M) and thermophilic starters (T), Lactobacillus plantarum ssp. plantarum ATCC 14917 (Lp), a mix of the M and T starters (M1), and mix of the M, T, and Lp starters (M2)—were used in the production of the goat cheeses. Volatile compounds were determined by a solid-phase microextraction/gas chromatography-mass spectrometric (SPME/GC-MS) method. The results showed that the moisture content of cheeses produced with the 5 kinds of starter cultures decreased after maturation, whereas ash content increased. The pH values of goat cheeses decreased first and then increased during maturity, and the pH value of M2 cheese was the lowest among the cheeses. The hardness and chewiness of the cheeses increased with increasing maturity, whereas cohesiveness, springiness, and resilience showed the opposite tendency. The 60-d-old cheese made with Lp had the highest chewiness, cohesiveness, springiness, and resilience, whereas the 60-d-old cheese made with M2 had the highest hardness. A total of 53 volatile components were identified by SPME/GC-MS, and carboxylic acids, alcohols, ketones, and esters were the 4 major contributors to the characteristic flavors of the cheeses. Volatile components and their contents differed greatly among the produced cheeses. The M2 cheese contained the highest relative content of the main volatile compounds (90.10%), especially butanoic acid and acetoin. Through a comprehensive comparison of the results, we concluded that M2 cheese had a dense texture and milky flavor, and M2 is a potential starter culture candidate for the production of goat cheese.     

Changes in volatile compounds and related biochemical profile during controlled fermentation of cv. Conservolea green olives

The effect of controlled fermentation processes on the profile of volatile and other biochemical compounds of cv. Conservolea green olives processed by the Spanish method was studied. The different treatments included: (a) inoculation with a commercial starter culture of Lactobacillus pentosus, (b) inoculation with a wild strain of Lactobacillus plantarum isolated from a previous fermentation, (c) uninoculated spontaneous process (control). Microbial growth, pH, titratable acidity, reducing sugars, organic acids and volatile compounds were monitored. Starter cultures were effective in establishing an accelerated fermentation process. Both were able to reduce the survival period of Enterobacteria by 7 days, minimizing thus the likelihood of spoilage. Higher acidification of the brines and faster pH drop was observed in inoculated processes, with L. pentosus presenting better performance than the wild strain of L. plantarum. Lactic and acetic were the major organic acids detected by HPLC, the concentration of which increased in the course of fermentation. Citric and malic acids were also present in the brines but they were degraded completely within the first 2 weeks of fermentation. Ethanol, methanol, acetaldehyde, ethyl acetate, isobutyric acid were the major volatile compounds identified by GC. Their concentration varied greatly among the fermentation processes, reflecting varying degrees of microbial activity in the brines.