Comparison of the volatile composition and sensory characteristics of Spanish PDO cheeses manufactured from ewes’ raw milk and animal rennet

Idiazabal, Manchego, Roncal and Zamorano varieties are the Spanish cheeses manufactured under Protected Denomination of Origin (PDO) from ewes’ raw milk and coagulated with animal rennet. Two batches of each cheese variety were ripened for six months and the cheeses were compared for sensory characteristics and composition of volatile compounds. Seventy-six volatile compounds were identified by GC-MS analysis of headspaces of the cheeses. The volatile profile of the four PDO cheeses differed significantly. Acids were the most abundant volatile compounds in Idiazabal, Roncal, and Zamorano cheeses, whereas alcohols were the main volatile compounds in Manchego cheese due to the large percentage of 2-butanol. Aldehydes, ketones and esters were minor compounds in all the cheese varieties, whereas terpenes and unsaturated hydrocarbons were only found in Manchego cheese. The sensory profiles, of the four PDO cheeses also differed significantly. The highest scores for sharp, brine and rennet odours, and rancid and rennet flavours were assessed in Idiazabal and Zamorano cheeses, whereas milky and buttery odour scores were higher in Manchego, Roncal and Zamorano cheeses than in Idiazabal cheese. Principal component analysis was applied to sensory attributes and volatile composition of the cheeses. Differences in flavour and odour attributes were correlated with differences in the volatile compounds. Two principal components correlating sensory attributes and volatile compounds were defined as “strength factor” and “sweetness factor”. The “strength factor” distinguished among the four cheese varieties, except between Roncal and Manchego cheeses, and the “sweetness factor” distinguished the Zamorano cheese from the Idiazabal, Manchego and Roncal cheeses.     

Changes in lipid fractions and sensory properties of Idiazabal cheese induced by lipase addition

This work studied the addition of an adequate lipase to enhance lipolysis reactions and the development of piquant flavour and sharp odour in Idiazabal cheese, as an alternative to the use of lamb rennet paste. Cheeses were manufactured from bulk raw ewes' milk in 50 l vats with commercial bovine rennet and 80 lipase units of pregastric or 180 lipase units of fungal lipase and ripened for 180 days. A higher lipolytic activity was induced by lipase addition promoting strong changes in odour and flavour attributes. Both fungal and pregastric lipases increased the content of total free fatty acids (FFA), but the fungal lipase released mainly medium- and long-chain FFA. In contrast, the pregastric lipase preferably released short-chain FFA. Diglyceride (DG) content was considerably higher in cheeses made with added pregastric lipase compared with those made with fungal lipase or with no lipase. Monoglycerides (MG) were detected only in cheeses made with either lipase added, reaching comparable concentrations after ripening for 180 days. The cheeses made with pregastric lipase had the highest scores for odour and flavour intensity, and sharp and rennet odours, desirable attributes for the Idiazabal cheese made with lamb rennet paste. None of the texture attributes were significantly influenced by the concentrations of MG and DG in the cheeses made with either lipase. Thus, the pregastric lipase was more appropriate than the fungal lipase to develop a more traditionally-flavoured Idiazabal cheese.     

Hygienic quality of ewes' milk cheeses manufactured with artisan-produced lamb rennet pastes

The use of artisan-produced lamb rennet pastes, but not any of the other commercial animal rennets, imparts a characteristic flavour to the cheese, so most Mediterranean ewes' milk cheeses are coagulated with this kind of rennet paste. In contrast to the advantages of using lamb or kid rennet pastes from the sensory point of view, questions are still raised as to their hygienic quality. The goal was to examine the microbiological and enzymic quality of lamb rennet pastes prepared by cheese manufacturers for their own use, and evaluate the hygienic quality of raw sheeps' milk cheeses made with them, using Idiazabal cheese as a model. Lamb rennet pastes prepared by artisan cheese makers from the Basque region of Spain (27), and Italy (8) were evaluated. For cheese making experiments 5 different lamb rennet pastes were selected among the 27 samples from the Basque Country region of Spain. Microbiological analyses were carried out on samples from rennet pastes, rennet extracts, milks and cheeses during ripening. Enzymic activities studied in rennet paste were: total coagulating strength and lipase. Analysis of variance and Student's t-tests was performed. The results show that the artisan-produced rennet pastes contain high levels of a variety of microorganisms. After 60 ripening days, which is the minimum ripening period required for Idiazabal cheese prior to its commercialization, no Eschericia coli, Clostridium, Salmonella spp. or Listeria monocytogenes were detected, and levels for the rest of the microorganisms were below the limits of the European legislative standards for cheese manufactured with raw milk. We can conclude that the use of artisan-produced lamb rennet pastes of questionable hygienic quality for the manufacture of raw milk hard cheeses yields products of good hygienic quality.     

Lipolysis,proteolysis and sensory properties of ewe’s raw milk cheese (Idiazabal) made with lipase addition

In this paper, we describe the effect of the addition of pregastric lipase on the composition and sensory properties of Idiazabal cheese. Free fatty acids (FFA), partial glycerides, free amino acids (FAA), gross composition and sensory characteristics were determined at different ripening times in cheeses manufactured with three different amounts of commercial animal lipase or with lipase-containing artisanal lamb rennet paste. The addition of lipase increased the content of total FFA, particularly of short-chain FFA, and that of total partial glycerides in cheeses. Unexpectedly, lipase utilization significantly affected total FAA concentration, which decreased in cheeses elaborated with high lipase amount. In general, Val, Glu and Leu were the major FAA, and their concentrations depended, mainly, on ripening time. Lipase addition had significant influence on the sensory characteristics of the cheeses, increasing scores for most of the flavour and odour attributes of the cheese. Principal component analysis (PCA) was done including dry matter, FFA, FAA, partial glycerides and odour and flavour attributes of the cheeses. It indicated that aroma and flavour parameters of Idiazabal cheese and the content of short-chain FFA and diglycerides were highly correlated to first principal component (PC1), while texture parameters, compositional variables and FAA were correlated to the second principal component (PC2).     

Volatile composition and sensory properties of industrially produced Idiazabal cheese

The volatile composition and sensory properties of industrially produced Idiazabal cheeses made from ewes’ raw milk (RM) or pasteurised milk (PM) and with addition of different starter cultures were compared. Cheeses were analysed at 90 and 180 d of ripening. Acids were the major volatile compounds in RM cheeses. Methyl ketones were the major volatile compounds in PM cheeses at 90 ripening days. However, the content of acids strongly increased with ripening whereas the content of ketones decreased in PM cheeses. The concentration of esters was higher in RM cheeses than in PM cheeses. No differences were found in the content of alcohols. Most aldehydes, hydrocarbons, terpenes and furans identified were minor volatile compounds in both RM and PM cheeses. In RM cheeses, characteristic sensory attributes for the aroma of Idiazabal cheese were present at 3 months, whereas in PM cheeses those desirable sensory attributes did not appear until 6 months of ripening.     

Short communication: Sensory profile of raw goat milk cheeses made with artisan kid rennet pastes from commercial-weight animals: Alternative to farmhouse goat cheeses

The loss of traditional kid rennet pastes in the Canary Islands (Spain), as in many other regions, is most likely due to the custom of using abomasa from very young animals killed below desirable commercial weight. In addition, the reasonable price of commercial rennets (CR) has resulted in the loss of typical sensory characteristics for most farmhouse raw goat milk cheeses, placing them at a disadvantage when local and international markets are full of different cheeses, often with aggressive marketing strategies. This paper analyzes the sensory characteristics of raw goat milk cheeses made with rennet pastes prepared from commercial kid abomasa in 2 ways: dried while full of ingested milk [full, commercial, artisan kid rennet (FCKR)], or dried after being emptied of ingested milk and refilled with raw goat milk [empty, commercial, artisan kid rennet (ECKR)]. This latter practice allows the use of empty abomasa, or abomasa with grass, soil, and so on. Sensory profiles of cheeses made with FCKR and ECKR rennets were compared with those made with CR by an expert panel (n = 7). The FCKR and ECKR cheeses had similar sensory profiles. Although scores for FCKR cheeses were somewhat higher than for ECKR cheeses, they were in the range found for traditional cheeses made with rennet prepared with abomasa from very young animals. The sensory profile of CR cheeses was very different. Almost 90% of consumer panelists (n = 90) preferred cheeses made with the experimental rennet pastes. These results demonstrate the possibility to prepare artisan rennet pastes from commercial-weight kids in an easy way for farmhouse cheese makers using local resources that would otherwise be destroyed in abattoirs.     

The effect of varying casein/fat ratio on physicochemical and sensory qualities of Feta‐type cheese made using buffalo milk

The Feta‐type cheese was prepared with different casein/fat (C/F) ratios of buffalo milk using microbial rennet. The manufactured Feta cheeses were subjected to physicochemical and sensory quality at 15‐day interval up to 60 days of ripening. Sensory analysis discriminated the different level of C/F ratio of buffalo milk cheeses predominantly by age. There was no significant difference (P < 0.01) observed in cheese made from C/F ratio of 0.6–0.7 in terms of flavour. The titratable acidity (TA), soluble protein and free fatty acid appear to be age‐dependent and increased throughout the ripening in all experimental cheeses.     

Short communication: To what extent do environmental or technological conditions affect the sensory differentiation of raw ewe milk cheeses produced in valley or mountain farms?

This work discusses the extent to which differences in technological or environmental conditions affect the sensory properties of cheeses made in mountain or valley farms. The work is focused on Idiazabal cheese, frequently made with lamb rennet paste and with milk from sheep flocks managed under extensive grazing systems. Fourteen sensory attributes were assessed, and free fatty acid content and rennet lipase activity were analyzed in cheeses from mountain and valley farms. Sensory differences between cheeses were mainly dependent on the type and amount of rennet used for cheesemaking, particularly on the level of rennet lipase activity. The sensory attributes did not appear to be influenced by pasture type, grazing practices, or environmental conditions associated with farm location. Rennet lipase activity was responsible for the high scores of strong sensory attributes such as pungent mouthfeel, and these attributes masked the potential sensory differences that could be found due to farm location.     

Effect of curd washing on cheese proteolysis,texture, volatile compounds,and sensory grading in full fat Cheddar cheese

Curd was washed to varying degrees during Cheddar cheese manufacture, by partial replacement of whey with water at the early stages of cooking, to give target levels of lactose plus lactic acid in cheese moisture of 5.3 (control), 4.5, 4.3 and 3.9% (w/w). The cheeses were matured at 8 °C for 270 days. While curd washing had little effect on composition or the mean levels of proteolysis (as measured by pH 4.6 soluble nitrogen and levels of free amino acids), it led to cheeses that were, overall, firmer and less brittle. Curd washing resulted in cheeses having lower levels of some volatile compounds, and being less acid, more buttery, sweeter, saltier and creamier than non-washed cheeses that had more 'sweaty', pungent and farmyard-like sensory notes. The results suggest that curd washing during Cheddar manufacture may be used as a means of creating variants with distinctive flavour profiles.     

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.     

Effect of artisanal kid rennet paste on lipolysis in semi-hard goat cheese

This study examined the use of hygienised kid rennet pastes in model cheese systems and also in goat milk semi-hard cheeses to promote lipolysis. The results obtained indicated that the use of rennet paste caused greater lipolysis and increased, mostly, the short-chain free fatty acid (FFA) content. The model systems made with whole goat’s milk using rennet paste and commercial rennet mixture exhibited a higher FFA content than did the rennet paste-free controls (31,600 vs. 25,600 μmol/kg cheese). For the pilot cheeses made with bovine rennet and rennet paste mixture, the increase in FFA level after 45 days of ripening compared with the cheeses prepared only with commercial rennet was as much as 6600 (μmol/kg cheese) and the increase in the butyric acid content was also 1650 (μmol/kg cheese). Moreover, after 15 days of ripening, industrially prepared cheeses made with rennet paste exhibited greater levels of FFA than did the cheeses made with commercial rennet (11,500 μmol/kg at 45 days of ripening). Their flavour was stronger and the organoleptic characteristics were better accepted, which implies less ripening time for commercial cheese manufacture.     

Changes in the free amino acid content during ripening of Idiazabal cheese: influence of starter and rennet type

The effect of starter and rennet type on free amino acid release during ripening of Idiazabal cheese was studied. Four batches of cheeses were manufactured depending on the rennet used, commercial calf rennet or artisanal lamb rennet, and the addition or not of starter culture. Cheese samples contained 24 individual free amino acids Leu, Glu, Val and Phe showing the highest contents during the ripening. The results indicated that the release of the free amino acids during ripening was strongly affected by starter added to the cheeses, and that this effect varied markedly with the rennet used for cheesemaking. Total amounts of free amino acids were higher for the cheeses made with commercial calf rennet than for those made with artisanal lamb rennet, regardless of starter addition. Likewise, the highest total free amino acid levels were found in the cheeses made with starter, regardless of the type of rennet used.     

Influence of ripening temperature on the volatiles profile and flavour of Cheddar cheese made from raw or pasteurised milk

Cheddar cheeses were made from raw (R1, R8) or pasteurised (P1, P8) milk and ripened at 1°C (P1, R1) or 8°C (P8, R8). Volatile compounds were extracted from 6 month-old cheeses and analysed, identified and quantified by gas chromatography-mass-spectrometry. A detailed sensory analysis of the cheeses was performed after 4 and 6 months of ripening. The R8 cheeses had the highest and P1 the lowest concentrations of most of the volatile compounds quantified (fatty acids, ketones, aldehydes, esters, alcohols, lactones and methional). The R8 and P8 cheeses contained higher levels of most of the volatiles than R1 and P1 cheeses. Ripening temperature and type of milk influenced most of the flavour and aroma attributes. Principal component analysis (PCA) of aroma and flavour attributes showed that P1 and R1 had similar aroma and flavour profiles, while R8 had the highest aroma and flavour intensities, highest acid aroma and sour flavour. The age of cheeses influenced the perception of creamy/milky and pungent aromas. PCA of the texture attributes separated cheeses on the basis of ripening temperature. The R8 and P8 cheeses received significantly higher scores for perceived maturity than P1 and R1 cheeses. The P1 and R1 cheeses had similar values for perceived maturity. In a related study, it was found that concentrations of amino acids and fatty acids were similar in R1 and P1 during most of the ripening period, and R1 and P1 cheeses had low numbers of non-starter lactic acid bacteria (NSLAB). The panel found that ripening temperature, type of milk and age of cheeses did not influence the acceptability of cheese. It is concluded that NSLAB contribute to the formation of volatile compounds and affect the aroma and flavour profiles and the perceived maturity of Cheddar cheese.     

Effect of partial or total substitution of bovine for caprine milk on the compositional,volatile, non-volatile and sensory characteristics of semi-hard cheeses

Semi-hard cheeses were manufactured from caprine milk and bovine milk mixed in ratios of 100:0; 75:25; 50:50; 25:75; 0:100 (caprine:bovine) and ripened at 12 °C for 150 d. Substitution of up to 75% bovine for caprine milks had no significant effect on levels of cheese protein, moisture-in-non-fat substances, salt, pH, whiteness, total or individual free amino acids and substitution of up to 50% had no significant effect on levels of primary proteolysis and free fatty acid and fruity flavours. Increased bovine milk content resulted in decreased levels of butanoic to octanoic acids, ethyl and methyl esters of acetic to octanoic acids and acyclic, mono and bicyclic terpenes and waxy/goaty flavours to a degree dependent on the proportion of each milk type. Partial substitution of caprine with bovine milks resulted in cheeses with chemical and sensorial characteristics that may provide an available alternative to consumers desiring caprine cheeses.     

Effects of pectic fat mimetics and transglutaminase on the regularity of protein and fat degradation and flavour compounds in Cheddar cheese during ripening

The effect of simultaneous transglutaminase (TGase) treatment with pectic fat mimetics (PFM) addition on regularity of protein and fat degradation and flavour compound variations in Cheddar cheese during ripening was investigated. In the early stages of fermentation, the cheese with 20% PFM and 15 U/L TGase had a higher amino acid content. In the middle stage of fermentation, cheese with high concentration of TGase hydrolysed more proteins. Cheese supplemented with PFM and TGase showed increased content of long-chain polyunsaturated fatty acids, especially linoleic acid (C18:2). In addition, PFM and TGase compensated the flavour deficiencies of low-fat cheeses and had positive effects on volatile compounds such as alcohols, acids and methyl ketones. Overall, the characteristics and flavours of Cheddar cheese with the addition of PFM and TGase were superior to the control group, which could provide a theoretical basis for the application of PFM and TGase in cheese production.     

Lamb rennet paste in ovine cheese manufacture. Lipolysis and flavour

In a preliminary study with commercial ewe's milk cheeses, there were statistically significant differences in the sensory evaluation scores and the amounts of short-chain (C4–C10) free fatty acids (FFAs) between cheeses made with lamb rennet paste or bovine rennet. Experimental ewe's milk cheeses were manufactured with 2 levels of artisanally produced lamb rennet paste and 2 levels of bovine rennet in 50 L vats, with a manufacturing replicate done within 1 week. Total coagulating activity was 2500 RU for cheeses manufactured with a high amount of rennet or 1000 RU for cheeses manufactured with a low amount of rennet. The two batches of lamb rennet pastes used had 4.0 (early Spring) and 6.5 U g⁻¹ lipase (late Spring), whereas no lipase activity was detected in bovine rennets. The total concentration of FFAs in cheeses manufactured with lamb rennet paste was significantly higher than in cheeses manufactured with bovine rennet, regardless of ripening time and time of the year. Lipolysis in the former cheeses increased with the total units of lipolytic activity added. The increase in lipolysis in cheeses made with lamb rennet was primarily due to higher amounts of short-chain fatty acids (C4–C10). Butyric acid was the main FFA in cheeses made with lamb rennet paste, representing 34.5 μmol 100 μmol⁻¹ (low level of rennet) and 44.2 μmol 100 μmol⁻¹ (high level of rennet) of the total FFAs, whereas it represented only 17 μmol 100 μmol⁻¹ of the total FFAs in all cheeses made with bovine rennet. The concentration of individual FFAs of chain length <C12 were significantly higher in cheeses made with lamb rennet paste than in cheeses made with bovine rennet. Cheeses made with lamb rennet paste received significantly higher intensity scores for odour and flavour intensity, sharp odour, ‘piquant’ and bitter flavours and ‘natural rennet’ odour and flavour.     

Evaluation of Harp Seal Gastric Protease as a Rennet Substitute for Cheddar Cheese

A crude preparation of gastric proteases from Harp Seal (Pagophilus groenlandicus) was found to coagulate milk over a wider pH range than porcine pepsin and had a higher ratio of milk clotting to proteolytic activity with hemoglobin at pH 1.8. Cheddar cheese prepared with seal gastric protease (SGP) gave significantly higher sensory scores than cheese made with calf rennet. Chemical analysis of the cheeses revealed a lower concentration of citrate-HCl soluble nitrogen and less free and peptide-bound amino acids in SGP cheese than in the cheeses made with calf rennet and Mucor miehei protease.     

Relationships between flavour and microbiological profiles in Serra da Estrela cheese throughout ripening

Cheeses manufactured in certified dairies in the Portuguese region of Serra da Estrela, using refrigerated and non-refrigerated raw sheep milk, were quantitatively evaluated in terms of indigenous microflora and volatile compounds during a ripening period of up to 6 mo. Viable counts were obtained for lactococci, lactobacilli, leuconostoc, enterococci, yeasts, Enterobacteriaceae and staphylococci; analyses of volatiles were performed by chromatography after solid-phase micro-extraction. Treatment of all analytical data produced by principal component analysis revealed correlations between the major microbial groups present in cheese and patterns of volatiles generated. End products resulting from the degradation of sugars, free amino acids and glycerides constituted the predominant volatiles of Serra da Estrela cheeses. Among volatile, short-chain carboxylic acids detected were acetic, propionic, iso-butyric and iso-valeric acids; these compounds are known to be breakdown products of Gly, Ala and Ser, of Thr, of Val, and of Ile, respectively, following oxidative deamination. Semi-volatile fatty acids and their corresponding ethyl esters appeared in the cheese, probably as a result of the activity of lipases produced by yeasts and Enterobacteriaceae. These ethyl esters, which are responsible for fruity flavours, were especially pronounced in cheeses manufactured from refrigerated milk.     

Contribution of lactic acid bacteria to flavour compound formation in dairy products

Cheese flavour cannot be produced without starter bacteria. Lactic acid bacteria convert lactose to lactic acid and this together with their production of diacetyl and acetaldehyde are their main contributions to the flavour of cultured milks and fresh cheeses. In matured cheeses, the starter bacteria die out quickly and the rate at which they lyse and release their enzymes into the system has an influence on the rate at which free amino acids are formed. Rennet alone is mainly responsible for the formation of large, medium and small peptides but, without interaction with other enzymes, is capable of producing only methionine, histidine, glycine, serine and glutamic acid at quantifiable levels. Free amino acids in Cheddar cheese are mainly the result of microbial peptidase activity. These amino acids, together with the products of glycolysis, form substrates for secondary flora, the nature of which, in many cases, determines the cheese variety. They also form substrates for enzymes from the milk, e.g. the production of H₂S appears to be dependent on milk enzymes. Methionine, which is released by rennet, is further metabolized by starter enzymes with the production of methanethiol which plays a major role in cheese flavour possibly as a potentiator for other flavours. —Dicarbonyls, particularly methylglyoxal and diacetyl, and bacteria which can produce them, appear to play a crucial role in the formation of cheese flavour, both the desirable flavour of full-fat cheese and the meaty-brothy off-flavour of low-fat cheese. Although, theoretically, there are many compounds in cheese which could react purely chemically to form flavour compounds, these reactions are also mediated by enzymes in the cheese system and it seems unlikely that straight out chemical reactions play a major role in the production of cheese flavour. The role of the secondary flora is likely to be much more important than that of chemical reactions. Particularly in Cheddar and Emmental it has been shown that good quality cheeses have a low oxidation-reduction potential. This is more likely to be an indicator for the establishment of the anaerobic conditions required for the flavour forming reactions to proceed than an active causal agent of flavour formation. The function of glutathione is more likely to be as some sort of facilitator in enzyme reactions than as an agent for the reduction of oxidation-reduction potential. The ability of bacteria to accumulate glutathione from their media is likely to be one of the indicators of flavour generating capacity. Suitable selected strains of adjunct bacteria increase the rate and intensity of formation of Cheddar cheese flavour but unsuitable adjuncts can also cause off-flavours.