Ultraviolet-absorbing compounds in milk are related to forage polyphenols.

The aim of this work was to characterize UV-absorbing compounds (UAC) in milk in relation to diet. In winter, 4 groups of cows each received a different diet: concentrate rich containing 35% cocksfoot hay (CCH), maize silage (MS), rye grass silage (RS), or rye grass hay (RH). In summer, 2 additional diets were given: mountain grassland hay (GH) and mountain grassland pasture (GP). Polyphenols were analyzed by HPLC and Folin reaction on forages and UAC were extracted from milks and analyzed by HPLC. In forages, the number of polyphenols was lowest in MS (57) and greatest in GP (85). Twenty-four peaks were identified, accounting for 28 to 47% peak area at 280 nm. Caffeoyl compounds and flavonoid glycosides were mainly found in RH, GH, and GP. Hydrolyzed compounds such as hydroxycinnamic acids and aglycones were found in MS and RS. Estimated amounts of polyphenols were lowest for MS (3.7 g/kg), roughly similar for CH, RS, and RH (about 15 g/kg), and greatest for GH and GP (21.6 and 35.3 g/kg, respectively). About 230 different peaks were separated in milks. Milks from RH and GP contained the lowest (87) and the greatest (127) numbers of peaks, respectively. Only 10 peaks were identified, accounting for 21 to 54% of the total spectra area. In addition to the major compound hippuric acid, phenylacetic acid, benzoic acid, 4-hydroxybenzoic acid benzaldehyde, catechol, and small amounts of ferulic acid were found in varying amounts depending on the diet. Flavonoids such as quercetin, luteolin, and apigenin were also present. Hippuric acid was clearly related to the presence of caffeoylquinic compounds in forages. Other identified UAC may originate essentially from forage simple polyphenols or from cell wall aromatics. Some of the several unknown compounds may also originate from the transformation of other nutrients. Estimated amounts of UAC were widely variable within each animal group. They were surprisingly high in CCH and roughly similar in all milks from preserved forages (about 3.6 mg/L), with generally greater values for GH milks, whereas the greatest amount was found in GP milks (13.3 mg/L). Hierarchical clustering clearly discriminated the 6 diets, showing that there were major differences in GP milks. Some UAC were specific to one or a group of diets. Ultraviolet-absorbing compounds are therefore a potential tool to distinguish between milks according to diet. In addition, they may have a bioactive effect on milk component conservation or on human health.

Odor compounds in cheese made from the milk of cows supplemented with extruded linseed and alpha-tocopherol.

Three diets for cows were used to evaluate the effect of extruded linseed (EL) or extruded linseed plus alpha-tocopherol (ELVE) supplementation of a maize silage diet (CO) on the odor-active compounds of Saint-Nectaire cheese. Cheese odor and flavor profiles were studied by sensory analysis. The volatile compounds were extracted by purge and trap and separated by gas chromatography. The odor compounds were detected and identified using an 8-way olfactometric device and a mass spectrometer. Twenty-nine volatile compounds were considered as contributing to the odor of Saint-Nectaire cheese. Half the compounds identified were known to be lipid degradation products but not all of them were affected by the diet. Among the markers of unsaturated fatty acid degradation, hexanal was not affected, whereas heptanal was increased more by the ELVE diet (6 times) than by the EL (3 times) diet. The ELVE diet led to cheeses with butanoic acid and heptanal odor peaks that were, respectively, 2 and 6 times higher than with the CO diet, which explained the lower milk odor and flavor scores obtained by sensory analysis on ELVE cheese. Although the cheese-making date had a greater effect than the diet on the aromatic profiles of the cheese, principal component analysis showed that the differences between cheeses obtained on the 3 diets were repeatable. The EL diet successfully enhanced cheese nutritional value without noticeably changing its flavor. alpha-Tocopherol supplementation was found to be unnecessary, as no oxidized odor was found.

Kluyveromyces lactis but not Pichia fermentans used as adjunct culture modifies the olfactory profiles of Cantalet cheese.

Yeasts are commonly detected in cheese. Two yeast species, Kluyveromyces lactis and Pichia fermentans, were isolated at high populations from raw-milk Cantalet cheese, a French Protected Denomination of Origin hard cheese. To investigate the interest of these 2 species as adjunct cultures to promote flavor development of Cantalet cheese, they were added at 10(5) cfu/mL to microfiltered milk. The global microbiological, biochemical, and flavor changes induced by the presence of the yeasts in cheese were determined. Adjunct yeasts were present at 10(6) cfu/g in curd, declined to 10(4) to 10(5) cfu/g in cheese, and did not influence gross composition, content of free amino acids, or content of free fatty acids. By using 8-way gas chromatography-olfactometry in parallel with gas chromatography-mass spectrometry, 30 odorous compounds of Cantalet cheese were identified. The olfactory profiles of K. lactis cheeses contained significantly greater levels of 8 odorous compounds (ethanol, ethyl hexanoate, 4 aldehydes, and 2 branched-chain acids) compared with the control and P. fermentans cheeses. Sensory analysis of cheeses flavor discriminated K. lactis cheeses on only 2 attributes (acetaldehyde and alcohol odors). This study shows that yeast contribution is species-specific and that K. lactis, at a population of 10(6) viable cells/g, can influence Cantalet cheese flavor.

Novel multi-gas chromatography-olfactometry device and software for the identification of odour-active compounds.

The aim of this work was to devise a novel fully computerised chromatography-olfactometry system where eight sniffers detect volatile compounds obtained from a single chromatographic separation. The technical options taken to implement this system are described in detail. The new methods and software developed and applied to analyse data obtained by gas chromatography-olfactometry and mass spectrometry (GC-O/MS) are presented. We found that 46 odorous zones could be detected by a panel of eight sniffers during the 35min of the analysis of the volatile compounds extracted from a cheese by purge-and-trap. Synchronisation of olfactometry and mass spectrometry data enabled us to propose 44 structures to account for 44 out of these 46 odorous zones.

The effect of raw milk microbial flora on the sensory characteristics of Salers-type cheeses.

The sensory characteristics of Salers Protected Denomination of Origin raw-milk cheeses are linked to the biochemical composition of the raw material (milk) and to the resultant microbial community. To evaluate the influence of the microbial community on sensory characteristics, Salers-type cheeses were manufactured with the same pasteurized milk, reinoculated with 3 different microbial communities from 3 different filtrates from microfiltered milks. Each cheese was subjected to microbial counts (on selective media), biochemical tests, and volatile and sensory component analyses at different times of ripening. Adding different microbial communities to specimens of the same (biochemically identical) pasteurized milk lead to different sensory characteristics of the cheeses. Cheeses with fresh cream, hazelnut, and caramel attributes were opposed to those with fermented cream, chemical, and garlic flavors. The aromatic compounds identified (esters, acids, alcohols, and aldehydes) in these cheeses were quite similar. Nevertheless, one milk was distinguished by a higher content of acetoin, and lower 2-butanone and 3-methylpentanone concentrations. Over the production period of 1 mo, the different cheeses were characterized by the same balance of the microbial population assessed by microbial counts on different media. This was associated with the stability of some sensory attributes describing these cheeses. Nevertheless, there was no linear correlation between microbial flora data and sensory characteristics as measured in this study.

Roles of superoxide dismutase and catalase of Staphylococcus xylosus in the inhibition of linoleic acid oxidation.

Staphylococcus xylosus used as starter culture in sausages decreases the level of volatile organic compounds arising from lipid oxidation and so contributes to the aroma by avoiding rancidity. The aim of this study was to characterize the roles of catalase and superoxide dismutase (SOD) in the inhibition of free fatty acid oxidation by comparing antioxidant capacity of the S. xylosus wild-type strain with those of the katA mutant and the sod mutant. Antioxidant capacity was determined by measuring the volatile organic compounds and the conjugated diene hydroperoxides arising from linoleic acid oxidation. The three strains inhibited the oxidation of linoleic acid. However, the katA mutant, and especially the sod mutant, had less antioxidant capacity than the S. xylosus wild-type strain. Thus both catalase and SOD of S. xylosus contributed to the inhibition of lipid oxidation.

Prediction of Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus populations in yoghurt by Curie point pyrolysis-mass spectrometry.

We evaluated the potential of pyrolysis-mass spectrometry (PyMS) for quantifying the binary mixed population of Streptococcus salivarius subsp. thermophilus and Lactobacillus delbrueckii subsp. bulgaricus in yoghurt. For this purpose, a new analytical approach was developed. The yoghurt was transparised and its total bacterial population was recovered by centrifugation and estimated by turbidimetric measurement. The quantity of each population (L. bulgaricus, S. thermophilus) was then estimated in the pellet by PyMS, and the data were analysed by artificial neural networks (ANNs). In parallel, streptococci and lactobacilli were numerated on SYL agar and these data were used as reference values to predict the bacterial counts of each population by PyMS. A close correlation was established between the streptococci and the lactobacilli counts on SYL agar and PyMS measurements (r(2)=0.98 for S. thermophilus and r(2)=0.96 for L. bulgaricus). Combined turbidimetric measurement and PyMS/ANNs seemed to be a powerful method for obtaining rapid counts of binary mixtures of bacteria in yoghurt.

Characterisation of volatile compounds produced by bacteria isolated from the spoilage flora of cold-smoked salmon.

This study investigated the volatile compounds produced by bacteria belonging to nine different bacterial groups: Lactobacillus sake, L. farciminis, L. alimentarius, Carnobacterium piscicola, Aeromonas sp., Shewanella putrefaciens, Brochothrix thermosphacta, Photobacterium phosphoreum and Enterobacteriaceae isolated from cold-smoked salmon. Each bacterial group was represented by several strains. In addition, combinations of the groups were examined as well. Sterile blocks of cold-smoked salmon were inoculated, vacuum-packed and stored at 6 degrees C. After 40 days of storage at 6 degrees C, aerobic viable count and pH were recorded, the volatile fraction of the samples was analysed by gas chromatography-mass spectrometry (GC-MS), and spoilage was assessed by sensory evaluation. Among the 81 volatile compounds identified by GC-MS, 30 appeared to be released as a result of bacterial metabolism. Some of the effects of inoculated bacterial strains on the composition of the volatile fraction seemed to be characteristic of certain bacterial species. Sensory analysis showed relationships between bacteria, the composition of the volatile fraction and the organoleptic quality of smoked salmon.

Production of esters by Staphylococci.

The ability of resting cells and extracellular concentrates of Staphylococci to synthesize ethyl esters was studied in the presence of ethanol and short chain acids considered individually. All the strains synthesized ethyl esters, S. warneri was the highest producer and S. carnosus the lowest. Resting cells esterified preferentially butanoic acid, extracellular concentrates esterified butanoic, valeric and hexanoic acids. Acetic, decanoic and branched acids were poorly esterified. The activity of the extracellular concentrates with ethanol and butanoic acid was not modified by the pH (pH 5.5 or 7.0); but it was decreased at a temperature of 14 degrees C compared to 24 degrees C. For the resting cells it was the opposite, the activity was inhibited by acid pH and was not influenced by the temperatures. So the Staphylococci could produce esters during sausage manufacture.

Lipolysis, proteolysis and formation of volatile components during ripening of a fermented sausage with Pediococcus pentosaceus and Staphylococcus xylosus as starter cultures.

Bacterial growth, formation of acids, lipolysis, proteolysis, fat oxidation, formation of volatile compounds and flavour characteristics were followed during ripening and storage of a fermented sausage. The starter culture used was composed of Pediococcus pentosaceus and Staphylococcus xylosus. The number of Pediococcus sp. increased by 1.5 log cfu/g during the first day of processing and remained constant at this level for 3 weeks. The corresponding initial increase in the numbers of Staphylococcus sp., 0.4 log cfu/g, was followed by a rapid decrease in the viable numbers. Lactic acid, mainly d-lactic acid, and acetic acid were formed during ripening. The triglycerides were hydrolysed to 1,2-diglycerides and free fatty acids at the beginning of ripening, followed by the formation of 1,3-diglycerides and monoglycerides, indicating lipolytic activity. Moreover, the nonprotein nitrogen increased during ripening as a result of the proteolytic activity. Most of the changes with respect to pH, formation of d-lactic acid, acetic acid, peroxides and flavour development occurred during the initial 3 days of ripening, when growth of Pediococcus sp. and Staphylococcus sp. occurred. Lipolysis as well as proteolysis continued after this initial period. The volatile compounds identified belonged to several chemical families, viz. aliphatic and aromatic hydrocarbons, aldehydes, ketones, alcohols, phenols, carboxylic acids, esters, nitrogen compounds, sulphur compounds, chloride compounds, terpenes and furans. Many of the volatile compounds probably originated from smoke and seasoning (onion/garlic and pepper), while others were a result of the activities of muscle enzymes and bacteria.

Influence of pig crossbreed on the composition, volatile compound content and flavour of dry cured ham.

The extraction of volatile compounds from dry-cured ham using a dynamic head-space method coupled with gas chromatography mass-spectrometry (GC-MS) enabled the identification of a large number of components. Some of these compounds exhibited intense sensory characteristics during odour assessment. Using similar technology, the effect of pig crossbreeds: Landrace × Large-White, Duroc × Gascon-Meishan, Piétrain × Gascon-Meishan and Large-White × Gascon-Meishan, on both volatiles and the flavour of dry-cured hams was limited, although differences were observed in aromatic compounds like 1-octen-3-ol (mushroom), 2,3-butanedione and acetoin (butter like).

Effects of starter cultures on the biochemical characteristics of French dry sausages.

Bacterial starter cultures, consisting of lactic acid bacteria (Lactobacillus, Pediococcus) and staphylococci (Staphlylococcus carnosus. S. saprophyticus, S. warneri), have an important effect on the pH, lactate, acetate and free fatty acid contents of sausages. Sausages made with L. sake had the lowest pH whereas no change of pH was noticed in the controls and in the sausages inoculated with P. acidilactici. Inoculation of S. saprophyticus led to sausages with a high acetate content. Lipolysis occurred not only in inoculated samples but also in the controls, but it was the highest in the sausages inoculated with S. warneri.

Effects of starter cultures on the formation of flavour compounds in dry sausage.

The purpose of this work was to study the impact of starter cultures on the production of flavour compounds in dry sausages. The effect of six starter cultures corresponding to different combinations of lactic acid bacteria (Lactobacillus sake L110, Pediococcus acidilactici 725, P. pentosaceus 716) and different Staphylococcus species (S. carnosus 833, S. warneri 863, S. saprophyticus M31) strains were tested in a total of 30 dry sausages without spices. The analysis of flavour compounds using a dynamic headspace apparatus coupled to a gas chromatograph and a mass spectrometer enabled us to identify about 80 volatile compounds. They were of various origins-lipids oxidation, fermentations, amino acid catabolism and animal feedstuffs. The influence of the starters and especially the flavouring strains proved to have a major effect on the level of volatile compounds in dry sausages. The flavour tests led to more accurate determination of the sensory characteristics of important molecules in the flavour of dry sausages. The sensory analyses showed that the butter odour of dry sausages largely depends on the catabolism of carbohydrates and that curing and rancid odours were correlated with some typical compounds of lipid oxidation.

Diet and growth effects in panel assessment of sheepmeat odour and flavour.

The effects of sheep age and diet on several odours and flavours are described. Ram lambs raised on ewe's milk then a corn-based diet were compared with lambs raised on milk and a pasture of grass/clover, six treatments in all. A seventh treatment comprised very old ewes maintained on pasture. Fat and lean from forequarters was minced and cooked together. Cooked lean was assessed for intensity by a sensory panel for 10 flavour attributes. Four showed significant (P < 0.01) treatment effects: 'sheepmeat', 'animal', 'liver', and 'poultry'. Sheepmeat flavour was highest in the slow-grown pasture-fed lambs. Animal flavour-the flavour associated with the odour of confined livestock-showed a similar pattern with treatment. Liver flavour was highest in ewe meat, and the biochemical origin of this flavour is discussed. Eleven related odour attributes were assessed on the rendered fat with a novel olfactometer. Five attributes showed highly significant treatment effects for intensity (P < 0.001): animal and sheepmeat odours showed a similar distribution to the equivalent flavours; likewise cabbage and rancid odours were associated with the two slow-grown pasture treatments. A comparison of the odour and flavour statistics showed that the sense of smell was the more discriminating in sheepmeat assessment, and also confirmed that fat was the true source of sheepmeat odour/flavour. In respect of sheepmeat production for effective marketing, the data showed that at 90 days, a pastoral diet resulted in slightly enhanced odours when compared with a corn-based diet. By 215 days, however, many undesirable odours were exacerbated. Since these older rams were more sexually developed, a sex rather than an age effect could not be excluded. Rendered fat from this work was further used in a companion study (Yang et al., 1997. Meat Sci., 45, 183-200) in an attempt to link individual volatile compounds to odour attributes.

Relations between compositional traits and sensory qualities of French dry-cured ham.

The left hams of 20 pig carcasses were processed by dry-curing for either 179 (n = 10) or 273 days (n = 10). The Biceps femoris was taken at the end of the processing and submitted to chemical and sensory analyses. The following chemical traits were determined: pH, moisture, salt content, fatty acid content and composition of lipid classes (triglycerides, phospholipids, free fatty acids), nitrogen fractions (soluble and insoluble protein, non-protein nitrogen, free amino acids) and volatile compounds. Sensory analysis was performed by a panel of 12 trained members, who scored texture traits (firmness, dryness, fibrousness, mellowness), taste (salty, acid), intensity of aromas (typical of dry ham, fresh meat, fat, cured meat, rancid, metal), persistence of dry ham aroma, persistence of taste after deglutition, persistence of astringency, persistence of metallic taste. The sensory qualities of dry-cured ham showed noticeable changes between days 179 and 273. Firmness, dryness and intensities of aromas typical of dry-cured ham and cured meat were enhanced with time. Neither pH, moisture nor salt content played a noticeable role in determining the sensory quality of dry-cured ham. Among the volatile compounds, the levels of several ketones and 1-butanol were significantly correlated with the aromas of dry ham and cured meat, while rancid aroma was related to aldehydes, ethylacetate, 2,3-pentanedione and nonane. Therefore, the aromas of dry ham and cured meat appeared to be determined mainly by the products of lipid oxidation. However, they were related also to unidentified volatile compounds, whose identification would be of particular interest.

Fat-borne volatiles and sheepmeat odour.

The effects of lamb age and diet on volatiles from fat are described. Rendered fat from ram lambs raised on ewe's milk then a corn-based diet was compared with that from lambs raised on milk and a pasture of grass/clover, six treatments in all. An additional treatment comprised very old ewes maintained on pasture. Helium-borne volatiles of rendered fat were resolved on a DB5 gas chromatographic column and the mass spectra obtained. Long chain alka(e)nes like neophytadiene were dominant in pasture treatments especially where the lamb growth rate was slow. Branch chain fatty acids (4-methyloctanoic, 4-methylnonanoic and an unidentified acid) were also highest in these treatments. Longer chain aldehydes like 2-undecenal were good indicators of a grain diet. Hexanal, commonly associated with rancid odours, was unaffected by treatment. The diketone 2,3-octanedione was an excellent indicator of a pasture diet, as was 3-methylindole (skatole). Phenols showed complex relationships to treatments, but were generally more common in pasture treatments. Benzenethiol (thiophenol) was unaffected by treatment. Inspection of principal component analysis plots identified 10 volatile compounds as contenders for the cause of sheepmeat odour; branch chain fatty acids were confirmed as the leading chemical class. There were indications that puberty or age caused an increase in the odorous 4-methylnonanoic acid. Animal odour-the odour of confined livestock-was clearly causally linked to 3-methylindole, a rumen breakdown product of tryptophan. 3-Methylindole was also responsible for rancid odour, rather than hexanal and its analogues. A hypothesis is advanced that links 2,3-octanedione formation to the enzyme lipoxygenase and linolenic acid, both abundant in green leafy tissue. Overall, the data confirm that sheepmeat odour/flavour is specifically linked to the branch chain fatty acids, and is probably exacerbated by pasture-derived 3-methylindole and alkyl phenols.

Fat volatiles tracers of grass feeding in sheep.

Volatile compounds were analysed in subcutaneous fat from lambs raised and finished on grass (GG); raised and finished on concentrates (SS); raised on grass and finished on concentrates for a long (GS1) or a short (GS2) period. Whereas 3-methylindole (skatole), a compound that has been described as a good discriminator of grass feeding, was unaffected by the diet (P>0.05), 2,3-octanedione was lower (P<0.01) in the fat from animals that spent a period in stall with a concentrate diet than in those finished on grass. Among the 20 monoterpenes detected, only p-cymene was affected by the treatment, being higher (P<0.05) in the fat from animals raised and finished on grass (GG) than in the other treatments. Eight among the 13 detected sesquiterpenes were affected by the diet treatment. In particular ß-caryophyllene, not detected in the fat from animals raised and finished on concentrates (SS group), was at basal levels in the animals raised on pasture and finished for different durations on concentrates (GS1 and GS2 treatments) and was present at high levels in the animals raised and finished on grass (P<0.0005). From a factorial discriminant analysis, four of the 33 terpenes detected discriminated perfectly those animals raised and finished on pasture (GG) from all the other groups (SS, GS1, GS2).

Solid-phase microextraction-mass spectrometry: a new approach to the rapid characterization of cheeses.

This work describes a new method for the rapid characterization of cheeses by solid-phase microextraction coupled with mass spectrometry (SPME-MS). After four types of fiber were tested and the main extraction parameters studied, the volatile components were extracted using a Carboxen/PDMS 75-microm fiber placed for 10 min at 20 degrees C in the headspace of the cheese. The substances adsorbed were then transferred directly from the injector to the inlet of a mass spectrometer through a 1-m deactivated silica capillary column heated to 210 degrees C. The mass spectra thus obtained without prior chromatographic separation formed a "fingerprint" of the analyzed sample. For data analysis, the mass fragments of each spectrum (45 < m/z < 150 amu) were considered as potential descriptors of the composition of the headspace of the cheeses. Stepwise discriminant analysis was used to select a limited number of mass fragments that afforded an operational classification of the batches of cheeses studied. This new method offers the advantage of minimizing thermal, mechanical, and chemical modifications of the matrix, thereby reducing the risk of analytical artifacts. SPME-MS provides a simple and effective approach to rapid quality control by analysis of the volatile fraction of foods.

Effect of forage conservation (hay or silage) and cow breed on the coagulation properties of milks and on the characteristics of ripened cheeses.

Forty-two multiparous dairy cows of three different breeds (Holstein, Montbéliarde and Tarentaise) were fed on the same type of forage (natural grassland) preserved in the form of either hay (H) or silage (GS), according to a changeover design (two 4 week periods). The proportion of concentrate in the diet and the energy and nitrogen contents were similar in both treatments. The milk produced by these cows was used for the manufacture of Saint-Nectaire type cheeses, under controlled and identical cheesemaking technological conditions. More cheese was produced with the H treatment milk. The cheeses made with the GS treatment milk were more yellow and tended to be more bitter. The other chemical and sensory characteristics did not differ much between the two treatments. Of the 51 volatile compounds identified, four were in significantly higher proportion in the GS than in the H cheeses. Cheeses produced from Tarentaise cows' milk were more yellow and their pH was higher than those made with the milk of Holstein or Montbéliarde cows. The cheeses from Montbéliarde and Tarentaise cows' milk were firmer, more melting and tastier than those made with the milk of Holstein cows. Although some trends were apparent, there were no significant differences in cheese volatile compounds for different breeds.