Isolation of lactic acid bacteria from Lighvan cheese, a semihard cheese made from raw sheep milk in Iran

The lactic acid bacteria contributing to Lighvan cheese ripening during the different stages of production were investigated. Isolated strains from different culture media were identified phenotypically to species and subspecies level. In total, 413 strains were isolated from raw milk, 1-day-old cheese and fully ripened cheese. The most abundant species belonged to Enterococcus faecium (87 isolates), Lactococcus lactis ssp. lactis (68 isolates), Enterococcus faecalis (55 isolates) and Lactobacillus plantarum (48 isolates). E. faecium, Lc. lactis and Lb. plantarum were the predominantly isolated strains from ripened cheese. Therefore, they may contribute considerably to the aroma and flavour development of Lighvan cheese.     

Changes of lactose, lactic acid, and acetic acid contents in Serra cheese during ripening

 Changes in the quantities of lactose, lactic acid and acetic acid in Serra cheese were monitored using a triplicate two-way factorial design over a ripening period of 35 days (sampling at 0, 7, 21 and 35 days) throughout the cheesemaking season (sampling in November, February and June). The amount of lactose in total solids of cheese (TS) decreased slowly from 6.17% to 0.21% (w/w_TS) as ripening time elapsed. As a result of sugar metabolism, the lactic acid content increased from 0.07% at day 0 to 2.10% (w/w_TS) by 35 days, whereas the acetic acid content increased from 0.00% to 0.24% (w/w_TS) during the first week. The lactose content was statistically correlated with the lactic acid content but not with the acetic acid content. Received: 18 November 1996     

Behaviour of pesticides lindane and methyl parathion during manufacture, ripening and storage of feta cheese

The behaviour of the pesticides lindane and methyl parathion during the manufacture, ripening and storage of feta cheese was investigated. Pasteurization of milk at 63–65°C for 30 min did not affect the concentrations of either pesticide. Lindane and methyl parathion at concentrations of 0.10 mg/kg and 16.66 mg/kg of milk, respectively, did not affect the activity of the starter culture. The concentration of lindane in the cheese and in whey/brine did not change significantly during ripening (up to 240 days). The concentration of methyl parathion in the cheese increased progressively up to 120 days of ageing, but decreased significantly (P < 0.05) from 120 up to 240 days. Methyl parathion might be bound to casein, in particular to the seryl and phosphoseryl groups of casein. Changes of brine to decontaminate the feta cheese from methyl parathion and lindane residues did not affect their concentrations in the cheese.     

Physicochemical, microbiological and sensory changes during ripening and storage of Xinotyri, a traditional Greek cheese from raw goat's milk

Physicochemical, microbiological and sensory characteristics, evolution of lipolysis monitored by measuring acid degree value (ADV) and the main mineral elements were studied during ripening and storage of artisanal Xinotyri cheese from raw goat's milk. Cheeses were characterized by a high content in total solids (TS; 83%) and fat (59% of TS), and very low pH (4.0), water activity (0.87) and moisture (17%). Protein and salt contents at the end of storage were 31% and 2.8%, respectively. Lipolysis increased, while variations in mineral (Ca, P, Mg and Zn) content were found during ripening and storage. Cheeses were free of Salmonella and Listeria in 25 g, while enterobacteria, pseudomonads, and coagulase-positive staphylococci were < 100 cfu/g. Mesophilic lactic acid bacteria increased above 8 log cfu/g by day 6, but declined by 2–3 logs in the ripened cheese (45 days) and by 3–4 logs during cheese storage at 4°C for up to 180 days.     

Changes in the physicochemical,microstructural and rheological properties of traditional Kope cheese during ripening

The effect of ripening time on the microstructural, physicochemical and rheological characteristics of traditional Kope cheese ripened in clay pots was investigated. The moisture content, pH and total nitrogen (TN)/dry matter (DM) of the cheese decreased, and DM, fat in DM and water‐soluble nitrogen/TN increased during this period. Storage and loss moduli increased, while loss tangent decreased; as a result, the elasticity characteristics were greater than the viscous characteristics of the samples. Microstructure images showed that the size of pores and casein network density increased significantly during ripening and, after 90 days, the structure of many pores became very similar to each other.     

The effect of refrigerated storage of raw milk on the physicochemical and microbiological quality of Tunisian semihard Gouda‐type cheese during ripening

A Tunisian semihard Gouda‐type cheese made from milk kept at 4 °C for 24, 48, 72 and 96 h was monitored during 45 days of ripening. The effect of milk refrigeration on the evolution of physicochemical parameters in relation to the quantitative variation of the microbial population during ripening of Gouda‐type cheese was investigated. Microbiological and physicochemical analyses were performed on raw milk and cheese samples after curding, 2, 9, 16, 23, 30, 37 and 45 days of ripening time. The raw milk kept under refrigeration at 4 °C for 96 h showed the highest microbial count and proteolysis level. The duration of storage significantly reduced the cheese yield as a result of important solubilisation casein in proteoses‐peptones. Results of different nitrogenous fractions by Kjeldahl method showed enzymatic hydrolysis products of casein whose intensity depended on the maturing stage as well as the refrigeration time. Besides the evident action of the plasmin, original milk protease, on the hydrolysis of casein in soluble fractions, the proteolysis of cheese caseins is also initiated by proteolytic action of the chymosin and extracellular heat‐resistant proteases notably produced by the same psychrotrophic microflora. Lactic acid bacteria starters that constitute the dominant microflora of this type of cheese are also considered as aroma precursors.     

The biodiversity and evolution of lactic flora during ripening of the Iranian semisoft Lighvan cheese

Ninety‐five isolated strains of Lactic acid bacteria (LAB) were identified from Lighvan cheese. The LAB evolution showed the dominance of lactococci and lactobacilli in the first stage and substitution of these genera by enterococci at the end of ripening. The most predominant strains were Enterococcus faecium (22.44%), Lactococcus lactis ssp. lactis (20.4%), Lactobacillus plantarum (18.36%) and E. faecalis (14.28%), respectively. Eleven and 51 different carbohydrate fermentation profiles were observed according to API 20 STREP and API 50 CH, respectively. API 20 STREP dendogram showed identical fermentation profiles of some E. faecalis and E. faecium strains, indicating that these strains might be well adapted to the whole cheese manufacture.     

Biochemical changes during the ripening of homemade 'San Simón da Costa' raw milk cheese

'San Simón da Costa' cheese is a traditional smoked variety produced in the northwest of Spain from cow's milk. Biochemical changes were determined during its ripening. Its high calcium and phosphorus content and its low NaCl and sodium content stand out. This cheese undergoes moderate proteolysis. The most abundant free amino acid at the end of the ripening was glutamic acid, followed by tryptophan, leucine, arginine and phenylalanine. The lipolysis throughout ripening is slight; the most abundant free fatty acid being oleic, followed by palmitic and butyric acid.     

Compositional characteristics and volatile organic compounds of traditional PDO Feta cheese made in two different mountainous areas of Greece

The compositional characteristics of Feta cheese manufactured in two mountainous areas from raw milk, from sheep fed by grass, and the forages’ volatile organic compounds (VOC) were studied, to differentiate the Feta cheese. Although the main compositional features did not vary greatly, the proteolysis mode was altered. The VOC profiles of the cheeses varied. While some VOCs were common to both groups of cheeses, others were discriminatory for each production area. There were common terpenes in both the bulk forage and the mature cheeses. Overall, the VOC composition of Feta could provide a basis for identification with regard to its region of origin.     

Monitoring changes in feta cheese during brining by magnetic resonance imaging and NMR relaxometry

Magnetic resonance imaging (MRI) and NMR relaxometry were used to monitor changes in feta cheese during 169 h of brining at 4.8%, 13.0% and 23.0% salt solutions. Image and relaxation data were acquired to study salt uptake and water loss due to dehydration of cheese during brining. Saturation recovery and Carr-Purcell-Meiboom-Gill (CPMG) sequences were used to determine the longitudinal relaxation (T₁) and the transverse relaxation (T₂) times, respectively. Signal intensities of T₂ weighted images decreased during 169 h of brining. An excellent linear correlation between the average signal intensity and the water content was obtained (R² = 0.984). The T₁ values of cheese brined at 4.8% were almost constant but T₁ values decreased for both 13.0% and 23.0% salt brined cheeses. Analysis of the CPMG decays gave relaxation spectra containing two components which decreased during brining. The short component T_2a was highly correlated with salt content (R² = 0.974). Results showed that NMR and MRI can be used to follow salt uptake and changes in water content in cheese during brining.     

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).     

FTIR-based polyphasic identification of lactic acid bacteria isolated from traditional Greek Graviera cheese

This study used a combination of phenotypic, physical (Fourier Transformed Infra-Red [FTIR] spectroscopy) and molecular (RFLP and SSCP analysis of 16S rRNA genes) methods to identify the lactic acid bacteria (LAB) flora present in traditional Greek Graviera cheese after five weeks of ripening. A total of 300 isolates collected from high dilution plates of TSAYE (incubated at 30 °C), M-17 (22 °C) and M-17 (42 °C) agar media were clustered by FTIR and then representative strains of each cluster were cross-identified blindly by all methods. Based on their FTIR spectra, 282 isolates were LAB grouped in 28 clusters. The LAB species identified and their prevalence in the cheese samples were: Lactobacillus casei/paracasei (68.8%), Lactobacillus plantarum (19.5%), Streptococcus thermophilus (8.9%), Enterococcus faecium (2.1%), and Lactococcus lactis (0.7%). Also, Staphylococcus equorum (11 isolates), Corynebacterium sp. (5 isolates) and Brevibacterium sp. (1 isolate) were recovered from TSAYE. Comparative identification results showed that phenotypic and molecular methods were in mutual agreement as regards the LAB species identified. The present polyphasic identification approach based on rapid FTIR screening of 10-fold more isolates than a previous classical identification approach allowed or improved detection of few sub-dominant species; however the predominant LAB species in the cheese samples were the same with both approaches.     

Changes in the rheological properties of Iranian UF-Feta cheese during ripening

The frequency sweep test was used to evaluate storage modulus (G′), loss modulus (G′′) and loss tangent [tan (δ)] of Iranian UF-Feta cheese during ripening period (3, 20, 40 and 60 days). With development of ripening, storage and loss moduli increased at varying rates. The rate of increase in G′ was greater than that in G′′ resulting in a reduction in tan (δ). That is, storage modulus was dominant to loss modulus and as a result the elasticity nature was greater than the viscous nature of cheese samples. Due to the disruption of fat globules and proteolysis, protein matrix rearranged and formed a more compact texture containing aggregates of casein. Ripening did not influence the pH level and also the concentrations of dry matter, fat, salt, and total nitrogen in dry matter. However, water soluble nitrogen increased significantly (P < 0.05).     

A factory-scale application of secondary adjunct cultures selected from lactic acid bacteria during Puzzone di Moena cheese ripening

The lactic acid populations of 2 seasonal Puzzone di Moena cheeses made from winter and summer raw cow's milk were characterized at different ripening times. Lactic acid bacteria (LAB) were isolated on selective media and subjected to genetic typing and identification. The species most frequently found during ripening were Lactobacillus paracasei ssp. paracasei, Lactobacillus plantarum, and Pediococcus pentosaceus. The different strains recognized by random amplification of polymorphic DNA-PCR were characterized for their acidifying and proteolytic activities to select nonstarter LAB to be used as secondary adjunct cultures (SAC). For each of the 3 above species, a strain showing weak acidification and high proteolytic capacity was selected. The 3 strains (Lb. paracasei ssp. paracasei P397, Lb. plantarum P399, and P. pentosaceus P41) constituted a mixed SAC used at 2 levels of concentration (10³ and 10⁴ cfu/mL) in experimental cheese making at dairy factory-scale. The analysis of volatile organic compounds as well as sensory analyses showed that the preferred level of SAC inoculation was 10³ cfu/mL.     

Role of salt in Iranian ultrafiltered Feta cheese: Some textural and physicochemical changes during ripening

The effect of NaCl (0–4%) on the ripening of Iranian ultrafiltered (UF)‐Feta cheese was assessed over 120 days of ripening at 4 °C. Whey percentage, whey salt, whey pH, cheese pH and textural properties of hardness and cohesiveness were monitored, and experimental modelling performed using response surface methodology. Texture, pH and whey percentage were significantly affected by NaCl and ripening. The maximum whey of 22% was recorded at the end of ripening period. Texture of this cheese becomes harder during ripening confirming cheese pH and whey percentage being the major determining factors. Cheese samples were more elastic than viscous with cohesiveness values of 0.6–0.9.     

Secondary proteolysis in Serra cheese during ripening and throughout the cheese-making season

 Experimental Serra cheeses were manufactured from raw ewe’s milk and thistle flowers following a two-way factorial design. The content of nitrogen soluble in water (WSN), in 2% trichloroacetic acid (2% TCA-N), in 12% TCA (TCA-N) and in 5% phosphotungstic acid (5% PTA-N), and the pH and salt-in-moisture concentration were measured throughout the ripening period (sampling at 0, 7, 21 and 35 days) and the cheese-making season (sampling in November, February and May). Proteolysis in 35-day-old Serra cheese was quantitatively high [average values of 34.6% and 11.9% for WSN/TN (total nitrogen) and 2% TCA-N/TN, respectively], but qualitatively low (average values of 5.8% and 1.2% for 12% TCA-N/TN and 5% PTA-N/TN, respectively). The ratios WSN/TN and 2% TCA-N/TN were lowest for cheeses ripened in February, whereas the ratio 12% TCA-N/TN was highest for cheeses ripened in November. By 35 days of ripening, the average pH and salt-in-moisture concentration values were 5.2 and 4.8%, respectively. No correlation was found to occur between the measured pH or salt-in-moisture concentration and the values of soluble nitrogen fractions throughout the cheese-making season. Received: 14 February 1996/Revised version: 4 June 1996     

The potential for carbon dioxide formation during ripening of acid curd cheese

Inspired by recent reports on high concentrations of carbon dioxide in the atmosphere of ripening chambers used in acid curd cheese production, small-scale experiments were performed to systematically investigate sources of CO₂ formation. In a closed system with a ratio of cheese mass to air volume close to that in industrial scale, up to approximately 16% (v/v) CO₂ were observed within 3 d of maturation at 24 °C. Without addition of ripening salts (CaCO₃, NaHCO₃) the initial carbon dioxide formation was delayed, but maximum CO₂ levels were still much higher than admissible workplace concentrations. Control experiments with quarg, which was pasteurized for yeast inactivation, revealed that growth and activity of yeasts has to be considered as the most important source for carbon dioxide formation. The results of the study strongly point on the necessity of preventive measures for ensuring the safety of employees.     

A model to assess lactic acid bacteria aminopeptidase activities in Parmigiano Reggiano cheese during ripening

The aim of this work was to investigate in which phases of ripening of Parmigiano Reggiano cheese lactic acid bacteria aminopeptidases present in cheese extract could be involved in release of free amino acids and to better understand the behavior of these enzymes in physical-chemical conditions that are far from their optimum. In particular, we evaluated 6 different substrates to reproduce broad-specificity aminopeptidase N, broad-specificity aminopeptidase C, glutamyl aminopeptidase A, peptidase with high specificity for leucine and alanine, proline iminopeptidase, and X-prolyl dipeptidyl aminopeptidase activities releasing different N-terminal amino acids. The effects of pH, NaCl concentration, and temperature on the enzyme activities of amino acid β-naphthylamide (βNA)-substrates were determined by modulating the variables in 19 different runs of an experimental design, which allowed the building of mathematical models able to assess the effect on aminopeptidases activities over a range of values, obtained with bibliographic data, covering different environmental conditions in different zones of the cheese wheel at different aging times. The aminopeptidases tested in this work were present in cell-free Parmigiano Reggiano cheese extract after a 17-mo ripening and were active when tested in model system. The modeling approach shows that to highlight the individual and interactive effects of chemical-physical variables on enzyme activities, it is helpful to determine the true potential of an amino-peptidase in cheese. Our results evidenced that the 6 different lactic acid bacteria peptidases participate in cheese proteolysis and are induced or inhibited by the cheese production parameters that, in turn, depend on the cheese dimension. Generally, temperature and pH exerted the more relevant effects on the enzymatic activities, and in many cases, a relevant interactive effect of these variables was observed. Increasing salt concentration slowed down broad-specificity amino-peptidase C, glutamyl aminopeptidase A, proline iminopeptidase, and peptidase with high specificity for leucine and alanine. Interestingly, this variable did not affect broad-specificity aminopeptidase N and positively affected X-prolyl dipeptidyl aminopeptidase. The models elaborated varying pH, temperatures, and salt concentration and were a useful, low cost, and fast tool to understand the role of the main peptidases in the different phases of cheese ripening in relation to the major environmental factors influencing enzyme activity.     

Chemical composition and microbial evaluation of Argentinean Corrientes cheese

The chemical and microbial composition of an artisanal cheese made from raw cow's milk produced and consumed in the province of Corrientes (north-eastern Argentina) was evaluated using standard methods. Corrientes cheese has high moisture content (50–60%), normal protein and fat contents (21–27 and 22–26% respectively), and is low in salt (0.5–2.0% w/w). Microbial counts also varied significantly between samples (colony-forming units per gram ranges covering logs of 5–11), probably due to environmental contamination in the raw material. These results will help produce higher quality Corrientes cheeses with well-defined characteristics.     

Evaluation of isolated starter lactic acid bacteria in Ras cheese ripening and flavour development

Eighteen cultures of starter lactic acid bacteria with or without added adjunct cultures, isolated from Egyptian dairy products, were evaluated in experimental Ras cheese for flavour development. Chemical composition of experimental cheeses was within the legal limit for Ras cheese in Egypt. All cultures used in this study had no effect on chemical composition of Ras cheese. Very significant variations in free amino acids, free fatty acids and sensory evaluation have been found among the cultures used in Ras cheesemaking. The levels of free amino acids and free fatty acids were correlated well with flavour development in Ras cheese. Seven of the tested cultures produced acceptable flavour and texture of Ras cheese. The highest overall score of flavour intensity, flavour and texture acceptability were in cheese made using YY47 lactic culture in addition to adjunct culture of Lactobacillus helveticus, Lactobacillus paracasei subsp. paracasei, Lactobacillus delbrueckii subsp. lactis and Enterococcus faecium. This culture can be recommended for Ras cheese manufacture using pasteurized milk.