1 Laboratoire de recherches fromagères, INRA, 36, rue de Salers, 15000 Aurillac;2 Laboratoire d'adaptation des herbivores aux milieux, INRA, Theix,
63122 Saint-Genès-Champanelle;3 Domaine de la Borie, INRA, 15190 Marcenat;
4 Station de recherche sur la viande, INRA, Theix, 63122 Saint-Genès-Champanelle, France
(Received 28 March 1995; accepted 28 June 1995)
Summary - Dairy cows belonging to 3 different genetic types (Holstein, Montbéliarde and Tarentaise)were fed, according to a 3 x 3 latin square experimental design, 3 different types of forage (ryegrasssilage, cocksfoot regrowths and maize silage). The proportion of concentrate in the diet, and theenergy and nitrogen contents were similar between ail 3 feeding treatments. The milk yielded by thesecows was used in the manufacture of Saint-Nectaire cheeses, under controlled technology manufac-turing conditions which were identical from one production to another. Cheeses manufactured from grasssilage milk have a more yellow curd than cheeses made from cocksfoot regrowlh or maize silagemilk. Cocksfoot regrowlh milk cheeses are more sticky and less firm than the others; they achieve a globalscore which is 0.6 to 1 point higher (P < 0.05) than those of other cheese types. Cheeses manufacturedfrom milk yielded by Montbéliarde and Tarentaise cows were better rated than those made fromHolstein milk. The various reasons for these differences are discussed in relation to the chemicalcomposition of the feeds.
cheese 1Saint-Nectaire 1 cow breed 1 forage type 1sensory characteristic
Résumé - Effet de la nature de la ration de base et du type de vache sur les caractéristiquesdes fromages de Saint-Nectaire affinés. Des vaches laitières de 3 types génétiques différents(Holstein, Montbéliarde et Tarentaise) ont reçu, selon un dispositif expérimental en carré latin 3 x 3, 3natures de fourrage différentes (ensilage de ray-grass anglais, foin de seconde coupe de dactyle et ensi-lage de maïs). La part de concentré dans la ration, et le niveau des apports énergétiques et azotés ontété semblables dans les 3 traitements alimentaires. Le lait de ces vaches a été transformé en fro-mages de Saint-Nectaire, dans des conditions technologiques de fabrications contrôlées et identiquesd'une fabrication à l'autre. Les fromages réalisés avec du lait d'ensilage d'herbe présentent une pâteplus jaune que celle des fromages réalisés avec du lait de foin de seconde coupe ou d'ensilage de maïs.Les fromages au lait de foin de seconde coupe se différencient des autres par leur caractère collant et
524 1Verdier et al
leur moindre fermeté; ils obtiennent une note globale supérieure de 0,6 à 1point (P < 0,05) à cellesdes autres types de fromages. Les fromages réalisés à partir du lait de vaches Montbéliardes etTarentaises ont été mieux notés que ceux réalisés à partir du lait de vaches Holstein. Les différentesorigines de ces écarts sont discutées en relation avec la composition chimique des aliments.
fromage / Saint-Nectaire / race de vache / nature des fourrages / caractéristique sensorielle
INTRODUCTION
The characteristics of matured cheesesdepend upon a great many factors linked, onthe one hand, to the technology employedand, on the other hand, to the original char-acteristics of the milk. The majority of stud-ies conducted over the past few years onthe influence of milk production factors oncheese characteristics, concerned the effectof genetic factors (Iactoprotein variants) onchee se chemical composition (Schaar et al,1985; Marziali and Ng-Kwai-Hang, 1986;Rahali and Ménard, 1991; Nuyts-Petit et al,1991; Remeuf, 1993; Delacroix-Suchet andMarie, 1994). Only a few studies revealedthe presence of a relationship between milkprotein genetic variants and the sensorycharacteristics of cheeses (Marziali and Ng-Kwai-Hang, 1986; Nuyts-Petit, 1991; Marieand Delacroix-Suchet, 1994; Vassal et al,1994). The influence of diet on milk chemi-cal composition (Hoden et al, 1985; Sutton,1989; Grummer, 1991; Hoden and Coulon,1991) and its rheological characteristics(Grandison et al, 1984, 1985a; Vertès andHoden, 1989; Laurent et al, 1992; Colin et al,1993; Macheboeuf et al, 1993) has beenstudied extensively. Sut most studies failedto approach the characteristics of the cor-responding matured cheeses, or only didso partially (Grandison et al, 1985b; Gareland Coulon, 1990). Some authors have,however, recently displayed a specifie inter-est in the sensory characterization ofcheeses (Serdagué et al, 1988, 1990;Paléari et al, 1993; Chamba et al, 1994).Eisewhere, numerous empirical observa-tions have been made associating differ-ences in cheese quality with variations in
farming practices (Urbach, 1990), and recentstudies (Martin and Coulon, 1995) have pro-vided explanatory factors. The relative lackof experimental data surrounding this topicis mainly linked to the difficulty in correctlydifferentiating the particular effects ofupstream factors fram those Iinked tocheese manufacturing itself. This approachis, however, of utmost importance in thecase of "Appellation d'Origine Contrôlée"-labelled cheeses for which modifications oforiginal milk characteristics upon cheesemanufacturing are generally highly limitedif not forbidden.
The aim of this study was to describeand analyse the organoleptic characteris-tics of Saint-Nectaire cheeses manufacturedfrom milk yielded by cows belonging to 3different breeds fed 3 types of forage (hay,grass silage and maize silage) in a 3 x 3latin square design. This study is a directextension of the work published by Coulonet al (1995) concerning the effect of type offorage on the chemical composition andclottinq ability of milks, the latter having beenused in the manufacturing of the Saint-Nec-taire cheeses studied in this paper.
MATERIALS AND METHODS
Experimental design
The cheeses studied were manufactured from 9different batches of milk yielded by animais of 3genetic types (Holstein [HO], Montbéliarde [MO],Tarentaise [TA]), fed a diet based on cocksfootregrowths (R), ryegrass silage (GS) or maizesilage (MS). A total of 54 multiparous cows (18 per
Forage type, breed and cheese characteristics
genetic type) in mid lactation (the mean lactationstage at the beginning of the first experimentalperiod was 10 weeks) were used. The experi-ment was conducted according to a 3 x 3 latinsquare design over 3 experimental periods (eachof 3 weeks) during which each group of cowswas successively fed the 3 types of forage. The 9groups (3 groups per breed) were formed on thebasis of production performances during a pre-experimental period and of the genetic variant ofthe K casein. The quantities of forage and con-centrate (mixture of barley, soya-bean meal andurea) fed, were calculated so that energy andnitrogen contents were similar between ail 3 diettreatments. On average, forage representedbetween 67 (group MS) and 72% (group R) oftotal dry matter (DM) intake. The data concerningthe setting up of the groups and the diets of theanimais have been explained in detail elsewhere(Coulon et al, 1995).
Manufacturing technology
The cheese transformations were carried out in anexperimental mini-cheese dairy equipped with 2vats having a maximal capacity of 40 l, on theTuesdays, Wednesdays and Thursdays of thelast 2 weeks of each period. In each vat, 34 kg ofmilk were used, allowing the manufacturing of 2actual-size Saint-Nectaire cheeses per batch.Saint-Nectaire cheese was selected as the modelfor our study as it is an Auvergne cheese whichcarries the "Appellation d'Origine Contrôlée" andis small in size (21 cm in diameter, around 5 cmthick). It is manufactured exclusively with cows'milk originating from the area possessing theAppellation (a few communes of the Puy de Dômeand the Cantal). The cheese has a semi-hard,pressed, sally curd with a moulded rind. In eachperiod, 12 vats were transformed, that is, 24cheeses manufactured: each period thereforeallowed the repetition of 3 of the 9 batches.
The Saint-Nectaire cheeses were manufac-tured according to a technology used in the rnak-ing of dairy Saint-Nectaire with raw milk; the milksused were from the prior evening milking, refrig-erated in tanks and maintained at 4°C until trans-formation. The milks were first heated in the vatsto 32°C, by allowing the hot water f1uidat regulatetemperature to tlow in the jacket, then their fatcontent was standardized by total skimming of apartial volume, so as to obtain a fat contentlpro-tein content ratio close to 1.15. They were next
525
enriched with CaCI2 (0.15 g1kg) to compensate Calosses consecutive to chilling, and inoculated witha 1% culture with mesophilic Iyophilized starters(Flora Danica Direct, Sochal, St Etienne deChomeil, France) revitalized on 10% sterile recon-stituted skimmed milk. The milks were then ren-neted at 32°C with a 520 mg active chymosinproof rennet (Gand-gassiot, Granday Laboratory,Beaune, France), 36 9 of which were added per100 kg of milk. Clotting time was assessed visu-ally. Twenty-five min after renneting, the curdwas cut for 5 min, then stirred for 40 min afterextracting a part of whey. It was then left to drainin a pre-pressing tray until it reached a pH closeto 6.20. Two cheeses per vat were then formed incloth mou Ids and pressed until a pH between5.30 and 5.40 was attained. Once this pH hadbeen reached, the cheeses, identified by num-bering on a case in plate, were removed fromhoops and brine salted for 7 h at 10°C. They werethen placed in a ripening cellar for 8 weeks. Tem-perature in the cellar was between 10 and 12°Cand relative humidity between 97 and 98%.
Physicochemical analyses
The pH was measured by introducing an Ingoldelectrode into some ground-up cheese, the tem-perature of which had first been brought up to20°C. Dry matter content was determined bydessication at 103°C for 24 h. The fat content ofthe cheeses was measured by butyrometricmethod (FIL-IDF standard 152:1991). The yel-low index of the cheese curds was determinedusing a spectraphotometer equipped with an inte-grating sphere (Renerre, 1981), on both faces ofa disk cut from the center of the cheeses.
Sensoryanalyses
The characteristics of the cheeses were assessedin 2 steps, using 2 very different methods.
1) Ali of the matured cheeses were submitted toa tasting panel of 8 assessors belonging to theINRA laboratory of Aurillac. The quality of thecheeses was assessed according to the criteriaissued by the Commission de Contrôle du Syn-dicat du Saint-Nectaire: members of the panelgave a score for the appearance of the cheese(/5), the curd (/5) and the taste (/10).
526 1Verdier et al
2) ln addition, a cheese from each batch wassubmitted for tasting to a sensory analysis groupcomprising 10 subjects, ail volunteers, trained torecognize primary tastes (AFNOR standard 1988)and familiar with sensory analysis and its tools.This panel had initially been required to assessvarious characteristics (appearance, odour, taste,texture) of retail Saint-Nectaire. A product profilewas subsequently established by the free gen-eration of allributes whose numbers were reducedwith the agreement of the tasting panel (AFNORstandard 1990; Barthélémy, 1990). The subjectswere trained to assess the intensity of each ofthe allributes included in the final profile. Scor-ing was do ne on a structured scale from 0 to 10.The profile comprised 2 parts of 15 allributeseach: 1 involved the "by mouth" description ofthe product, the other concerned the "by appear-ance" description of the cheese. The assessorsalways began with the actualtasting: the cheesesamples, with rinds removed, were coded andpresented simultaneously to alltasters in a ran-dom order which was different for each person.Cheese appearance was assessed on a quarterof a cheese (different code from the former part):ail cheeses were presented simultaneously to ailassessors.
Analysis of volatile compounds
A total of 20 9 of cheese were used to analyse thevolatile compounds of 2 cheeses, chosen in the 2most different groups according to sensory anal-ysis. This analysis was performed by gas chro-matography-mass spectrometry, as described inBerdagué et al (1993), and used to detect differ-ences in the concentration of substances whichplay a role in the f1avour of cheeses (Bosset etal, 1994), and to put forward chemical and bio-chemical hypotheses as to the origin of thesesubstances.
Statistical analysis
Results were analysed using the mean scoresof both methods of assessment (tasting and sen-sory analysis). The data were processed by anal-ysis of variance (SAS, 1987), whilst including inthe model the forage type, breed, batch, the num-ber of the cheeses in the vat and the period, asweil as breed x treatment interaction.
RESULTS
For each group of animais, 2 cheeses weremanufactured, analysed and tasted. For ailparameters studied, these 2 cheeses wereidentical.
Effect of type of basal ration
The cheeses made from milk of animais fedregrowths (R cheeses) differed in certainaspects from cheeses made from grasssilage milk (GS chee ses) (table 1). Themaize silage milk cheeses (MS cheeses)generally showed intermediate character-istics. Cheese yield was higher, but not sig-nificantly, in MS cheeses than in GS and Rcheeses. This is linked to the fat and pro-tein concentrations of the milks used incheesemaking, which were slightly higherin the MS group. R cheeses were less drythan GS and MS cheeses (-1.1 points of drymatter, P < 0.05). Their yellow index wassignificantly lower than that of GS cheeses,but higher than that of MS cheeses (P <0.01). On tasting, their overall score washigher by, respectively, 0.6 (P < 0.05) and0.3 points (P < 0.05) than those of GS andMS cheeses.
By mouth, 2 attributes differed signifi-cantly (P < 0.01): R cheeses were moresticky and less firm than MS and ab ove ailGS cheeses. Overall acceptability score forR cheeses was greater by 1 point (P< 0.05)to that of GS cheeses and by 0.6 points(P> 0.05) to that of MS cheeses.
Effect of breed
The cheeses manufactured from the milkof ail 3 breeds of cow showed similarphysicochemical characteristics (curd colour,dry matter, pH) (table 1).The fat in dry mat-ter of the chee ses done on Tarentaises milk
Forage type, breed and cheese characteristics 527
Table 1. Effect of forage and animal breed on cheese characteristics.Effet de la nature du fourrage et de la race sur les caractéristiques du fromage.
1 GS: ryegrass silage; R: cocksfoot2nd eut hay; MS: maize silage. 2 HO: Holstein; MO: Montbéliarde;TA: Tarentaise.3 Residual standard deviation. 4 Alter partial skimming. 5 100 x [(curd weight x curd DM)/(milk weight x milk DM)].abc Within diet or breed, values with different superscripts within row differ significantly (P < 0.05).1 GS: ensi/age de ray grass anglais; R : foin de dactyle de seconde coupe; MS : ensilage de maïs. 2 HO : Holstein;MO: Montbéliarde ; TA : Tarentaise. 3Écart type résiduel. 4 Aprés écrémage partiel. 5 100 x [(poids de caillé xteneur en MS du caillé)/(poids de lait x teneur en MS du lait)].abc À l'intérieur de la race ou du régime, les valeurs suivies d'une lettre différente sont significativement différentes
(P < 0,05).
528 1Verdier et al
(TA cheeses) was, nevertheless, lower by1.6 points (P < 0.01) than those of HO andMO cheeses. However, HO cheeses weresignificantly different (P < 0.01) from thecheeses of the 2 other breeds (MO and TAcheeses) in ail tasting criteria (appearence,texture, taste). The total score was lowerby nearly 1 point for HO as compared toMO and TA.
ln the sensory analysis, no differencesbetween the 3 cheese types appeared sig-nificant, apart from the more melting char-acter of MO and TA cheeses (P < 0.05), butthe tendances observed confirmed the dis-parities highlighted by the panel of asses-sors. Global acceptability was thereforegreater by 0.2 and 0.5 points, respectively,for MO and TA cheeses comparatively toHO cheeses.
9.00
8.00
o...7.00
6.00
-;, 5.00c'§ 4.00oIII 3.00
The disparities in cheese characteristicsbetween breeds were noticeably differentaccording to the diet fed (P < 0.01). In par-ticular, MO cheeses achieved their besttaste score (13.9) with the regrowth dietwhereas TA cheeses achieved this (13.5)with the maize silage-based diet. The grasssilage-based diet resulted in the smallestdifferences between breeds. Finally, the dif-ference in taste score reached almost 2points between the cheeses made withMontbéliarde cows and cocksfoot regrowthsand those made with Holstein cows and rye-grass silage. The qualities differentiatingthese 2 groups of cheeses were essentiallyopening number, firmness and gritty textureof the curd (fig 1). The volatile compoundsdistribution was analysed on 1 cheese ofeach of these 2 groups (MO/R and HO/GS).
. \ \/ \~ \2.00
1.00
Fig 1. Sensory characlerislics of cheeses according 10 type of forage and cow breed .• HO/GS; 0 MO/A.Caractéristiques sensorielles des fromages selon la nature du fourrage et la race .• Holstein/ensilagede ray-grass anglais ..Q Montbéliarde/foin de dactyle de seconde coupe.
Forage type, breed and cheese characteristics
Volatile compounds
Forty-two molecules were identified by GC-MS measurements (table Il). Most of thesecompounds belonged to the following chem-ical families: alkanes or alkenes (10), alde-hydes (4), ketones (6), alcohols (5), esters(7) and aromatic hydrocarbons (3). Thecomparison between the chromatographicprofiles of HO/GS and MO/R cheesesrevealed a desorption 5 times superior inHO/GS cheese. The differences betweenthe 2 cheeses lay essentially in the sulphurcompounds whose contents were very lowor indeterminable in MO/R cheese. Thesecompounds, which present very low thresh-olds of olfactory perception and whoseimpact on cheese sensory characteristicsis important, originate probably from anadvanced microbial sulphur-amino acidcatabolism during the ripening phase(Hem me et al, 1982). Esters, some of whichare able to contribute to a "fruit y" note ofthe product, the 2-3 butanedione, which is atypical compound of glucidic catabolism witha butter odour, and many 2-methyl-ketoneswere identified in more important quantitiesin the HO/GS cheese. In contrast, the MO/Rcheese distinguished itself by a more impor-tant quantity of cyclic-alkenes. The oxida-tion degree of the 2 cheeses was limited asshown by the low desorption of aldehydes,alcohols and unramified aliphatic alkanes.
DISCUSSION
One of the difficulties in studying the actualeffects of production factors on the quality ofmatured cheeses lies in the control of thecheesemaking technology employed. In thisstudy, ail precautions were taken to ensurethat variations of technological order fromone production to another were as slight aspossible (use of identical vats, regular mon-itoring of pH leading to a placing in mouldand end of pressing conducted under sirnl-
529
lar conditions between productions). More-over, the latin square experimental designallowed the random distribution betweentreatments of possible variations caused bynoncontrolled modifications in the technol-ogy.
Effect of the type of forage
One of the main effects of the type of dietconcerned the colour of the cheese curd. Asalready observed by Garel and Coulon(1990), grass silage results in a more yel-low curd than regrowth or maize silage. Theyellow colour of the cheese curd is due tocertain milk carotenoids originating fromfoodstuffs and more or less absorbed withinthe rumen depending on cell wall fat con-tent (Hay, 1993). The positive effect of grasssilage is due both to the type of forage(maize displaying lower carotenoid contentsthan grasses [Knigh and Waghorn, 1993])and, comparatively to regrowths, to itsmethod of harvesting which avoids longexposure to Iight, a major cause ofcarotenoid destruction (Britton and Goodwin,1973).
From a sensory point of view, cocksfootregrowth led to cheeses which were moresticky and less firm than the others. This isno doubt partially Iinked to their lower drymatter, and to the slightly lower useful mat-ter content of the milks used in manufac-turing. However, Grandison et al (1985b),in Cheshire technology, observed no effectof milk chemical composition on cheesesand their texture. Regrowth cheeses weregenerally better rated and globally more sat-isfying than GS and MS products. It is mostlikely that the observed disparities were dueto minor components which were eitherspecific or present to a greater or lesserextent in each forage (sesquiterpenes, lac-tones, dimethyl sulphyde) as already sug-gested by certain authors concerning milk(Forss, 1993; Visser, 1993; Wilson, 1993)
530 1 Verdier et al
Table II. Volatile compounds of 2 extreme cheeses.Composés volatils de 2 fromages extrêmes.
1 Measured Kovats indices. 2 The reliability of the identification is indicated by the following symbols: a: mass spec-trum and retention time identical with those of an authentic sam pie; b: mass spectrum and Kovats indices in agree-ment with the corresponding literature data; c: mass spectrum consistent with spectra found in the Iiterature; d: ten-tative identification by mass spectrum.1 Indices de Kovats mesurés. 2 La validité de l'identification est précisée par les symboles suivants: a = spectre demasse et temps de rétention identiques à ceux d'un échantillon de référence; b = spectre de masse et indices de Kovatsen accord avec les données de la littérature; c = spectre de masse voisin de ceux observés dans la littérature; d =essai d'identification à partir du spectre de masse.
Forage type, breed and cheese characteristics
or cheese (Dumont and Adda, 1978;Dumont et al, 1981; Bosset et al, 1994).This study does not enable us, however, tospecify whether these components wereoriginally in the forages or linked to theirmethod of conservation in the form of silage.Indeed, the latter can lead, in certain cases,to the development of distinctive f1avours incheese (Urbach, 1990; Forss, 1993).
Effectofanirnalbreed
ln this trial, the Montbéliarde breed led tothe most favourable cheese characteristics,especially in comparison with Holstein,These differences must be interpreted withcaution. On the one hand, they can be dueto feed effects. Indeed, considering theexperimental protocol, ail animais receivedcomplements as required, which means thatthe greater the production level of the ani-mais, the more concentrated feed they weregiven. The cows belonging to group HOwere thus fed a diet comprising 40% of con-centrate against, respectively, 28 and 21 %for MO and TA animais. On the other hand,these disparities cannot be generalized to ailanimais of a breed, because our samplewas low and not representative. Our resultssimply show that differences in cheese qual-ity can be due to intrinsic animal character-istics. This type of observation has recentlybeen made for Beaufort type cheeses ortraditional caprine cheeses according to themilk ~ or us1 casein variants (Marie andDelacroix-Buchet, 1994; Vassal et al, 1994).Along the same lines, differences in colourand f1avour according to breed (Friesian vsJersey herds) have also been noted (Keenand Wilson, 1993).
CONCLUSION
This study is original in the fact that it wasconducted under controlled conditions of
531
milk production and cheesemaking. lt hasrevealed certain differences in cheese char-acteristics depending on type of forage andanimal breed, and thus confirms the numer-ous empirical observations made on thissubject (Urbach, 1990). These resultsshould, however, be confirmed by comple-mentary studies before being made gen-eral, in particular for breed effects. The dif-ferences observed were obtained withinthe context of a particular technology. Asinteractions exist between the effects of pro-duction factors and manufacturing technol-ogy employed (Martin and Coulon 1995), itis possible that the effects of production fac-tors might have been Iimited by the tech-nology used in this study (relatively rapidacidification of the curd). Moreover, thisstudy did not allow the separation of theeffect of the floristic type of forage itself fromthat of its method of conservation. Work ispresently underway to analyse the respec-tive specific effects of these 2 factors.
ACKNOWLEDGMENTS
We would like to thank R Lavigne (cheese trans-formation) and H Magne of the ENILV in Aurillac(sensory analysis) for their help, Y Vantome forthe cheese colour measurements, and the PôleFromager AOC Massif-Central with whom theparticipation of this study was performed.
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