Research Article Indigenous Starter Cultures to Improve ...

Research ArticleIndigenous Starter Cultures to Improve Quality ofArtisanal Dry Fermented Sausages from Chaco (Argentina)

Noelia Z Palavecino Prpich12 Marcela P Castro12 Mariacutea E Cayreacute1

Oscar A Garro12 and Graciela M Vignolo23

1Laboratorio de Microbiologıa de Alimentos Universidad Nacional del Chaco Austral Comandante Fernandez 755Presidencia Roque Saenz Pena 3700 Chaco Argentina2Consejo Nacional de Investigaciones Cientıficas y Tecnicas (CONICET) C1033AAJ Buenos Aires Argentina3Centro de Referencia para Lactobacilos (CERELA) Chacabuco 145 San Miguel de Tucuman 4000 Tucuman Argentina

Correspondence should be addressed to Marcela P Castro mcastrouncauseduar

Received 30 August 2014 Revised 17 December 2014 Accepted 14 January 2015

Academic Editor Sadhana Ravishankar

Copyright copy 2015 Noelia Z Palavecino Prpich et al This is an open access article distributed under the Creative CommonsAttribution License which permits unrestricted use distribution and reproduction in any medium provided the original work isproperly cited

Lactic acid bacteria (LAB) and coagulase negative cocci (CNC) were isolated from artisanal dry sausages sampled from thenortheastern region of Chaco Argentina In order to evaluate their performance in situ and considering technological featuresof the isolated strains two mixed selected autochthonous starter cultures (SAS) were designed (i) SAS-1 (Lactobacillus sakei 487 +Staphylococcus vitulinusC2) and (ii) SAS-2 (L sakei 442 + S xylosusC8) Cultures were introduced into dry sausage manufacturingprocess at a local small-scale facility Microbiological and physicochemical parameters were monitored throughout fermentationand ripening periods while sensory attributes of the final products were evaluated by a trained panel Lactic acid bacteria revealedtheir ability to colonize and adapt properly to the meat matrix inhibiting the growth of spontaneous microflora and enhancingsafety and hygienic profile of the products Both SAS showed a beneficial effect on lipid oxidation and texture of the final productsStaphylococcus vitulinus C2 from SAS-1 promoted a better redness of the final product Sensory profile revealed that SAS additionpreserved typical sensory attributes Introduction of these cultures could provide an additional tool to standardize manufacturingprocesses aiming to enhance safety and quality while keeping typical sensory attributes of regional dry fermented sausages

1 Introduction

InArgentina dry fermentedmeat products aremanufacturedby means of local artisanal techniques derived from Spanishand Italian traditions Artisanal production lacks standard-ization processes and relevant featuresmdashsuch as casing sizeraw materials quality and process parametersmdashare subjec-tively monitored [1] Moreover dry fermented meat productsare produced without the addition of starter cultures that isthe fermentation is conducted by wild-type strains henceensuring the development of the so-called indigenousmdashor autochthonousmdashmicroflora Under these conditions thesettlement of the same group of microorganisms givinga uniform fermentative behavior is an improbable sce-nario Consequently the introduction of starter cultures in

the manufacture of fermented meat products turns into anecessary implement in order to guarantee food safety andto standardize the final product attributes [2 3]

Meat fermented products are the result of a complexmicrobiological activity that mainly consists of a lacticfermentation and several characteristic biochemical changestriggered by lactic fermentation Meat starter cultures cur-rently comprise coagulase negative cocci (CNC) and lacticacid bacteria (LAB) Many CNC species such as Staphylococ-cus spp and Kocuria spp have nitrate reductase proteolyticand lipolytic activities which contribute to the developmentof redness and flavor of the product [4ndash7] As regards LABspecies mainly used as starter cultures are Lactobacillus sakeiL curvatus L plantarum L pentosus L casei Pediococ-cus pentosaceus and P acidilactici [8] Lactic bacteria play

Hindawi Publishing CorporationInternational Journal of Food ScienceVolume 2015 Article ID 931970 9 pageshttpdxdoiorg1011552015931970

2 International Journal of Food Science

a crucial role in this fermentation that is acidification whilethey can also have proteolytic and lipolytic activities Thepositive technological features of this acidification include (i)inhibition of spoilage and pathogenic microflora (ii) quickdrying and enhancement of texture due to denaturation andcoagulation of proteins (iii) activation of muscle proteasesand (iv) formation of nitric oxide and nitrosomyoglobinwhich contribute to redness [9] However an excessive acidi-fication can negatively affect the taste or smell of the productand lead to color failures as a consequence of CNC inhibition[10] The most important factors determining characteristicsand quality of meat fermented products are the selectionof starter cultures and the environmental conditions duringfermentation and ripening [11 12] Quality of the productswill be the result of the interaction between these two factorswhich generates a beneficial or a disadvantageous setting forcellular metabolism

Indigenous microorganisms from meat fermentation arewell adapted to fermentation and ripening conditions and areable to inhibit spontaneous growth of undesirable microflora[8] Selected autochthonous starter cultures (SAS) can con-tribute to the manufacture of artisanal local products withparticular features In previous studies two strains of L sakeione strain of S vitulinus and another of S xylosus wereisolated from regional meat fermented products [13] Owingto their safety and technological features these strains havepotential application as starter cultures In order to evaluatetheir performance in situ and consequently the feasibilityof their use these strains were tested as starter cultures in alocal small-scale facility which manufactures dry fermentedsausages Two different mixed starter cultures were designedMicrobiological and physicochemical characteristics of theproducts had beenmonitored throughout the manufacturingprocess while sensory attributes of the final products hadbeen evaluated by a trained panel

2 Materials and Methods

21 Autochthonous Starter Cultures Strains used in this studywere selected in previous studies taking into considerationtheir technological and safety features [13] they are reportedin Table 1 Association of the different strains to design eachstarter culture was determined by means of compatibilitytests Microorganisms are deposited in the Strain Repositoryof the Laboratorio de Microbiologıa de Alimentos (Universi-dad Nacional del Chaco Austral Argentina) Lactobacilli arekept in MRS broth (de Man Rogosa Sharpe) supplementedwith 20 vv glycerol while staphylococci are kept in soytrypticase + 06wv yeast extract (TSBYE) added with20 vv glycerol both bacterial species aremaintained frozenat minus18∘C

Two mixed starter cultures were designed namely SAS-1 comprised of L sakei 487 and S vitulinus C2 and SAS-2comprised of L sakei 442 and S xylosus C8 Lactic bac-teria were propagated in MRS broth and CNC strains inTSBYE they were incubated at 30∘C for 12 h Cells were thenharvested by centrifugation (4500 rpm 20min) washedtwice with saline peptone water (085 gNaCl100mL) and

Table 1 Indigenous bacterial strains used as starter cultures in thisstudy

Strain Technological activity

Lactobacillus sakei 442 Acidifying proteolytic and nitratereductase


Staphylococcus vitulinus C2 Proteolytic and nitrate reductaseStaphylococcus xylosus C8 Lipolytic and nitrate reductase

resuspended in this solution Cellular density was monitoredby plate count (MRS agar for LAB and Mannitol salt agar forCNC)

22 Dry Fermented SausagesManufacture and Sampling Dryfermented sausages used in this work were those currentlymanufactured by a local small-scale facility Traditionalrecipes dictate the composition of these meat products perkilogram that consists of 450 g beef 177 g pork meat 300 gbacon 34 g sodium chloride 15 g sugar 10 g powder milk07 g potassium nitrate 3 g sodium glutamate 5 g choppedgarlic 2 g ground red pepper and 3 g ground black pepperAll the ingredients were thoroughlymixed together andmeatdough was divided into two batches one batch was addedwith the SAS and the other was kept as a control systemAfterwards meat dough was packed and shaped into naturalcasings (sheep gut) SASwere added once themeat dough hadbeen finished being completely homogenized before casingswere stuffed Cultures SAS-1 and SAS-2were added separatelyto the meat dough in two different instances Final bacterialconcentration (LAB + CNC) reached a value of sim106 colonyforming units per gram (cfug) After inoculation and stuff-ing the unfermented products were placed in a ripeningchamber for 15 days During the first seven daysmdashferment-ation periodmdashtemperature was kept at 18ndash22∘C and relativehumidity (RH) oscillated to 90ndash95 The next 8 daysmdashripening periodmdashtemperature was kept at 15∘C andRH rangewas 80ndash85 From each batch samples were withdrawn toperform physicochemical and microbiological analyses atinitial time 2 4 7 and 15 days

23 Microbiological Analyses Ten grams of each sample wasaseptically taken and was transferred to a stomacher bag with90mL of sterile peptone water to be homogenized After2min in the stomacher (Stomacher 400 Circulator SewardUK) appropriate dilutions of the homogenate were done inorder to inoculate several growth media to determine viablecounts of LAB in MRS agar (30∘C 72 h) Micrococcaceaein AMS (30∘C 72 h) yeasts and molds in ChloramphenicolSabouraud Agar (25∘C 5 days) Enterobacteriaceae in Glu-cose Red Violet Bile Agar (37∘C 24 h) Staphylococcus aureusin Baird Parker Agar with egg yolk emulsion and potassiumtellurite (37∘C 48 h) Besides final productswere also assayedfor the following Escherichia coli count (MPNg) follow-ing the ICMSF method [14] sulphite-reducing anaerobes(cfug) according to ISO 152132003 [15] absence of E coli

International Journal of Food Science 3

O157 H7NM by the USDA-FSIS method [16] absence ofSalmonella spp by the BAM-FDA method [17] absence ofListeria monocytogenes according to ISO 11290-1A1 [18]

24 Physicochemical Analyses

241 Water Activity (119886119908) pH and Humidity Water activity

was measured at 25∘C by means of a water activity device(Testo Germany) Measurements of pH were determinedwith the aim of a pricking probe attached to a pH meterinserted directly into the meat dough (Testo) Humiditycontent (expressed as ) was determined according to theAOACmethod [19]These parameterswere evaluated at everysampling stage

242 Ash NaCl Concentration and Fat Content Ash contentwas analyzed according to the AOAC method [20] NaClconcentration and fat content were determined according toAOAC [19] These measurements were solely conducted onthe final product

243 Lipid Oxidation Determination Lipid oxidation wasevaluated by means of the thiobarbituric acid reactivesubstances test (TBARS) according to the methodologysuggested by Sinnhuber and Yu [21] TBARS values wereexpressed as mg malonaldehyde per kilogram of sample(mgMDAkg)

244 Free Fatty Acid (FFA) Composition The FFA compo-sitions were determined in the fat fraction of dry sausagesamples by means of gas chromatography of their corre-sponding methyl esters Briefly total lipid contents wereextracted according to Bligh and Dyer [22] and then theywere saponified esterified and transferred to n-heptaneas proposed by Hartman and Lago [23] Fatty acid methylesters were analyzed using an Agilent 6850AHP (AgilentTechnologies Inc CA USA) chromatograph equipped witha capillary fused silica column Supelco 2340 (Supelco Belle-fonte PA USA) (60m length 025mm bore and 025 120583mfilm thickness) splitndashsplitless injector and flame ionizationdetector The latter two were kept at a constant temperatureof 250∘CHeliumat 06 psi was the carrier gas Temperature ofthe oven was set at 140∘C for 5min followed by an increase to240∘C at a rate of 4∘Cmin The final temperature was keptconstant for 20min FFA were identified by comparison oftheir retention times with those of authentic standards(Supelco 37 Components FAME Mixture Bellefonte PA)FFA concentration was expressed as mg fatty acid per gramof fat

245 Color Measurements Color measurements were con-ducted in dry sausages cuts with the aim of an Evolution600UV-Vis spectrophotometer (Thermo Scientific equippedwith integrator sphere andVISIONLite Color Calc SoftwareGermany) illuminant D65 and a 2∘ standard observer in aroom with fluorescent lighting and after standardization ofthe instrument with respect to the white calibration plateCIE 119871lowast 119886lowast and 119887lowast values were determined as indicators of

lightness redness and yellowness Two sausages from eachtreatment were cut into 3 pieces The color of each piece wasmeasured at 5 locations on the covered cross section Themean of 15 measurements was recorded for the color of thedry meat sausage

25 Sensory Analysis A quantitative descriptive analysis(QDA) was conducted by an eight-member sensory panelwith previous experience in judging fermented sausages Fullripened dry fermented sausages manufactured with the SASand those used as control systems were evaluated Two 30-minute-long training sessions were heldmdashin which commer-cial fermented sausages were usedmdashso as to define discussand clarify each attribute to be evaluated The overall assess-ment was performed following Baka et al [2] guidelinesThe evaluated attributes were as follows (i) external appear-ance (descriptors firmness cohesiveness and easy peelingcapability of the case from the sausage) (ii) color (intensityand uniformity) (iii) aroma (intensity cured smell andrancidity) (iv) taste (salty acid and sour) (v) texture (chew-ing and hardening) The answer for every descriptor wasdetermined as the mean value of panelistsrsquo answers

26 Statistical Analyses All data were expressed as means plusmnstandard errors of duplicate measurements Statistical com-parisons between variables were performed with Studentrsquos 119905-test Differenceswere considered significant at119875 lt 005 One-way ANOVA was used to determine significant differences(119875 lt 01) between sensorial attributes All the tests were per-formed with the software Statgraphics Plus 40 (ManugisticsInc USA)

3 Results and Discussion

31 Fermentative Bacteria and Associated Microflora Lacticbacteria molds and yeasts Micrococcaceae and Enterobac-teriaceae counts are depicted in Figure 1 S aureus was notdetected in any of the samples taken during the productiveprocess

Lactic bacteria comprised the dominant microflorathroughout fermentation and ripening periods At the begin-ning of the productive process LAB counts in the inoculatedsystems were higher (SAS-1 588 plusmn 018 log cfug and SAS-2 626 plusmn 025 log cfug) than those of the control systems(C-1 215 plusmn 001 log cfug and C-2 429 plusmn 001 log cfug)Differences found in bacterial counts from both controlsystems reflected the high variability between batches thatcan be found in the processing plant During the first 48 hSAS-1 LAB counts rapidly increased to 864 plusmn 002 log cfugfollowed by a slight reduction to 827 plusmn 016 log cfug at theend of the fermentation period with no significant variationsuntil the end of the ripening period As regards SAS-2 LABcounts reached a value of 782plusmn016 log cfug at 48 h and thisvalue was kept invariable throughout the entire process LABcounts from inoculated SAS-1 showed significant differencescompared to the corresponding control system C-1 until thefourth day of fermentation whereas LAB counts from inoc-ulated SAS-2 showed significant differences compared to its


1

2

3

4

5

6

7

8

9

0 5 10 15Processing time (days)

Fermentation Ripening

log 1

0(c

fug

)

(a)

1

2

3

4

5

6

7

8

9


log 1

0(c

fug

)

(b)

0

1

2

3

4


log 1

0(c

fug

)

(c)

0

1

2

3

4


log 1

0(c

fug

)

(d)

Figure 1 Bacterial counts of (a) lactic acid bacteria (b) Micrococcaceae (c) yeasts and molds (d) Enterobacteriaceae throughoutfermentation and ripening periods Systems X-X SAS-1 (Lactobacillus sakei 487 + Staphylococcus vitulinus C2) ◼---◼ C-1 (noninoculatedcontrol system simultaneously manufactured with SAS-1) 998771- sdot -998771 SAS-2 (L sakei 442 + S xylosus C8) e sdot sdot sdote C-2 (noninoculated controlsystem simultaneously manufactured with SAS-2)

control system C-2 until the second day of fermentation Inaddition LAB counts of both control systems had been alsoincreased during the fermentation period reaching similarvalues compared to the inoculated systems at the end ofthe ripening period The LAB imminent colonization inthe systems inoculated with both SAS confirmed the goodadaptation of indigenous LAB strains to the meat substratesuggesting a high dominance and adaptability to the specificenvironmental conditions of the manufacturing processSimilar results had been reported by Baka et al [2] Bonomoet al [3] and Talon et al [24] among others

Micrococcaceae counts revealed no significant differ-ences between inoculated systems (SAS-1 and SAS-2) andtheir respective control systems (C-1 and C-2) in any of thesampled stages These results which had also been reportedby other authors [25 26] can be explained in the view thata great load of this microbial group could inhabit theprocessing environment facilitating its introduction into themeat dough during mixing and handling NeverthelessMicrococcaceae are weak competitors for nutrients com-pared to LAB though their countswere not higher than LABrsquoscounts Regarding molds and yeasts counts they did not

show significant differences between inoculated systemsand their noninoculated control systems during the studiedperiod Enterobacteriaceae showed bacterial counts that didnot differ significantly between SAS-1 and C-1 nor betweenSAS-2 and C-2 (mean values were 335 plusmn 006 and 220 plusmn018 log cfug resp) Similar counts for this microbial groupwere encountered in meat fermented products from otherregions [2 3 27] Bacterial counts from this groupdiminishedsignificantly at the second day of fermentation in system SAS-1 while this reduction was evident for SAS-2 only after thefourth day However in the different systems (inoculated andnoninoculated) bacterial counts were less than 1 log cfug atthe end of the ripening period (Figure 1(d)) The latterbehavior is expected since Enterobacteriaceae are highly sen-sitive to acidity and dryness [28] In general Gram-negativebacteria and Enterobacteriaceae are considered unwantedmicroflora in meat fermented products the dominance ofLAB and Micrococcaceae being an essential event on themanufacture of these products Besides quality control of thefinal products showed that all the samples were within thedesirable values for good hygiene and food safety of this typeof products namely E coli (MPNg) lt 3 sulphite-reducing


4

5

6

7

0 5 10 15

pH

Processing time (days)

(a)

20

30

40

50

60

70

0 5 10 15

Moi

sture

()


(b)

75

80

85

90

95


aw

()

(c)Figure 2 Physicochemical parameters determined during dry sausages fermentation and ripening periods (a) pH values (b) humidity() (c) 119886

119908

() Systems X-X SAS-1 (Lactobacillus sakei 487 + Staphylococcus vitulinus C2) ◼---◼ C-1 (noninoculated control systemsimultaneously manufactured with SAS-1) 998771- sdot -998771 SAS-2 (L sakei 442 + S xylosus C8) e sdot sdot sdote C-2 (noninoculated control systemsimultaneously manufactured with SAS-2)

anaerobes lt 10 cfug Salmonella spp absence in 25 g ofsample Listeriamonocytogenes absence in 25 g of sample andE coli O157 H7 absence in 65 g of sampled products

32 Physicochemical Parameters The results of pH 119886119908 and

humidity measurements during fermentation and ripeningof dry sausages are reported in Figure 2 Initial pH valuesin the meat matrix were not significantly different betweenSAS-1 and C-1 nor SAS-2 and C-2 (mean values 606 and572 resp) Furthermore no significant differences werefound between pH values of SAS-1 systems and their controlsystems throughout the productive process Values for SAS-2showed a significant reduction after two days of fermentationcompared to the control system (54 and 57 resp) howeverthis difference disappeared on subsequent days pH valuesbecame similar (sim547) at the end of the fermentation periodComparing SAS-1 to SAS-2 the first contributed to a deeperacidification of the fermented system that is SAS-1 produceda pH falling of 081 units while SAS-2 just decreased pH in02 units This reduction is due to carbohydrate fermentationwithin the meat matrix mediated by LAB which produces

organic acids responsible for pH decrease This situation inmeat fermentation is crucial since it contributes to the inhi-bition of undesirable microflora accelerates the developmentof redness affects flavor and reduces water holding capacityof proteins contributing to the necessary drying process [29]A slight pH increase in all the systems (with the exceptionof system C-2) was recorded during the ripening process itcan be attributed to the deamination effect caused by yeasts[30 31] and to the buffering capacity of proteins [32] Ingeneral pH values found herein are in agreement with thosefound by other authors [3 26 27]

Humidity values at the end of the ripening period rangedbetween 2758 and 3241 in all the samples (inoculatedand noninoculated) No significant differences were foundbetween SAS and control systems humidity values in any ofthe different stages of the productive process Similar resultswere reported by Palavecino et al [33] and Romero et al [34]for products manufactured in the same geographical regionas well as Casaburi et al [27] andCasquete et al [35] for Euro-pean products Nevertheless these values were considerably


Table 2 Ash and NaCl concentration total fat content and TBARSvalues registered for dry sausages at the end of the ripening period

System Ash () NaCl () Total fat () TBARS(mgMDAkg)

SAS-1 69a plusmn 031b 611 plusmn 097 2497 plusmn 429 089 plusmn 003C-1 642 plusmn 015 539 plusmn 038 2146 plusmn 267 105 plusmn 008SAS-2 695 plusmn 032 573 plusmn 018 3005 plusmn 262 097 plusmn 004C-2 638 plusmn 112 603 plusmn 001 3399 plusmn 055 154 plusmn 004aMean valuebStandard deviation

lower than others reported for products from other regionswhere for longer ripening periods humidity contents werehigher [3 7 12 36 37]119886

119908showed high values at the beginning of the process

(9265ndash9335) and then it diminished progressively to finalvalues ranging between 796 and 819 for all the systemsNo significant differences were found between inoculatedsystems and their respective controls

Other physicochemical parameters evaluated on finalproducts are presented in Table 2 As regards ash contentNaCl concentration and lipid content no significant differ-ences were determined between inoculated and noninocu-lated systems On the contrary TBARS values were higherin control systems C-1 and C-2 compared to systems SAS-1 and SAS-2 respectively Natural casings used to stuff drysausages are permeable to air and consequently sausages areexposed to lipid oxidation TBARS values were significantlyhigher in control system C-2 and from these results it couldbe inferred that SAS are able to prevent lipid oxidation in drymeat sausages and they restricted TBARS values to a concen-tration of 1mgMDAkg which is considered the ranciditylimit for fresh meat [38] Differences found amongst SAScould be attributed to their different antioxidant propertiesnamely metal chelating capacity free radical scavengingand reducing properties Although TBARS values close to2mgMDAkg had been found in fermented meat products[39 40] and even higher amounts (22mgMDAkg) theywere not enough to be detected as rancid at a sensory level[41]

Changes in color parameter are shown in Table 3 Resultsshowed that there were significant differences in the rednessin those samples inoculated with SAS-1 compared to theircontrol systems (C-1) Redness in fermentedmeat products isdue to nitrosomyoglobin formationwhen nitrogenmonoxidereacts with myoglobin Nitrosomyoglobin increases expo-nentially at the beginning of the drying process as a result ofnitrate reductase activity from staphylococci [42] MoreoverMicrococcaceae have the ability to produce the enzyme cata-lase which destroys the oxygen peroxide produced by LABOxygen peroxide affects heme pigments and can produceoxidative decoloration of dry fermented sausages From theseresults it could be inferred that the nitrate reductase positivestrain S vitulinusC2 (part of SAS-1) was effective in achievinga better redness development with the aim of the lowerpH found in these samples compared to the SAS-2 ones

Table 3 Color determination in dry sausages at the end of theripening period

System 119871

lowast

119886

lowast

119887

lowast

SAS-1 4067a plusmn 089b 999 plusmn 066 586 plusmn 037C-1 4135 plusmn 057 910 plusmn 055 555 plusmn 053SAS-2 3904 plusmn 105 853 plusmn 108 538 plusmn 062C-2 3868 plusmn 236 836 plusmn 076 490 plusmn 027aMean valuebStandard deviation

Regarding the rest of the color parameters no significant dif-ferenceswere found between the inoculated systems and theircontrol systemsOn account of fat content hematic pigmentsnitritenitrate concentration and humidity among otherfactors which affect reflectance values from these prod-ucts different chromatic values had been reported by manyauthors [43ndash45]

33 FFA Profile Free fatty acids concentration (expressed asmgg of fat) was determined in order to evaluate SAS effecton lipid fraction of studied meat products Samples manu-factured with the addition of SAS showed a similar lipidicprofile to the noninoculated samplesThese results are shownin Table 4 Talon et al [24] and Casaburi et al [27] describedsimilar tendencies Significant differences for oleic acid con-centration were found in system SAS-1 whose strains did notshow lipolytic activity in vitro compared to its control systemC-1This fact could be attributed to endogenous lipases activ-ity being favored by the pH decrease [46] On the other handlipolytic strain S xylosus C8 (part of SAS-2) was not effectivein the lipolysis of the products during ripening thoughendogenous lipases activity can be confirmed [47 48] Freefatty acidsmdashreleased as a consequence of endogenous lipasesand microbial activitymdashcomprise the substrate for thosebiochemical reactions that lead to aromatic compoundsformation and flavor of the final product [3 49 50]

34 Sensory Attributes Sensory quality of dry meat sausages(texture color and flavor) depends on numerous compoundsinvolved in many biochemical reactions that take place dur-ing themanufacturing process Final firmness of the productsis the result of its acidification and dryness Both eventsare favored by lactic acid bacteria owing to their acidifyingcapacity which also contributes to the dryness of the productbecause it reduces the water holding capacity of the mixture[51] In this study an enhancement in the firmness of inocu-lated dry sausages was observed which can be attributed tothe L sakei strains acidification activity from both SASSamples inoculated with SAS-1 (Figure 3(a)) also showedhigher punctuation for the following descriptors easy peelingcapability of the case from the sausage color intensity anduniformity The higher color intensity is in keeping withthe higher redness found in those samples inoculated withSAS-1 On the other hand samples inoculated with SAS-2(Figure 3(b)) obtained lower scores for the descriptor easypeeling capability of the case from the sausage No significantdifferences in the rest of the evaluated attributes were


Table 4 Free fatty acid (FFA) profile [mg fatty acidg of fat] of dry sausages manufactured with indigenous starter cultures

Free fatty acid SystemSAS-1 C-1 SAS-2 C-2

Saturated(140) myristic 1093a plusmn 099b 955 plusmn 084 1199 plusmn 301 1138 plusmn 215(160) palmitic 17816 plusmn 139 15275 plusmn 163 18985 plusmn 908 17891 plusmn 775(170) heptadecanoic 329 plusmn 043 296 plusmn 049 266 plusmn 057 257 plusmn 044(180) stearic 9225 plusmn 658 7605 plusmn 643 8749 plusmn 201 1814 plusmn 218(220) behenic 832 plusmn 070 730 plusmn 21 560 plusmn 164 507 plusmn 160

Monounsaturated(161) palmitoleic 1793 plusmn 146 1551 plusmn 165 1851 plusmn 331 1778 plusmn 228(171)c heptadecanoic 314 plusmn 035 257 plusmn 047 242 plusmn 042 237 plusmn 020(181)c n9 oleic 33187 plusmn 592 25706 plusmn 437 30102 plusmn 139 30552 plusmn 103

Polyunsaturated(182)c n6 linoleic 7941 plusmn 457 6907 plusmn 1005 7435 plusmn 1217 7336 plusmn 1053120572(183) n3 heneicosanoic 998 plusmn 068 897 plusmn 065 817 plusmn 164 765 plusmn 157(203)c n3 11417-eicosatrienoic 128 plusmn 026 129 plusmn 025 ND ND

aMean valuebStandard deviationND non detected

01234567

Easy peeling

Cohesiveness

Aroma intensity

Cured smellRancidity

Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

Color intensitylowast

Coloruniformitylowast

capabilitylowast

(a)

01234567

Easy peelingcapability

Cohesiveness

Color intensity

Coloruniformity

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

(b)

Figure 3 Effect of autochthonous starter cultures on the sensory attributes of fermented sausages (a) Comparison between SAS-1 and itscontrol systemC-1 (b) Comparison between SAS-2 and its control systemC-2 SystemsX-X SAS-1 (Lactobacillus sakei 487 + Staphylococcusvitulinus C2) ◼---◼ C-1 (noninoculated control system simultaneously manufactured with SAS-1) 998771- sdot -998771 SAS-2 (L sakei 442 + S xylosusC8) e sdot sdot sdote C-2 (noninoculated control system simultaneously manufactured with SAS-2) lowastSignificantly different (119875 lt 01)

observed compared to the control systems confirming thatthe designed SAS could contribute to enhance the qualityof these traditional local products without changing theirtypical sensory attributes

4 Conclusions

Native starter cultures represent an efficient alternative tocommercial starter cultures since the latter do not alwaysoffer the desirable properties to the final product Selectedautochthonous starters enhanced safety and hygienic profileof dry sausages showing also a beneficial effect on lipidoxidation and texture of the final products Sensory profile

revealed that SAS addition helped to preserve typical sensoryattributes inasmuch as any of the SAS affected it significantlyFinally the addition of these cultures could provide an addi-tional tool to standardize manufacturing processes aiming toenhance safety and quality while keeping typical sensoryattributes of regional dry fermented sausages Further studiesare being conducted in order to test the robustness of bothSAS in different production batches throughout the year

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper


Acknowledgments

The authors would like to thank the following institu-tions from Argentina for their financial support ConsejoNacional de Investigaciones Cientıficas y Tecnicas (CON-ICET) Agencia Nacional de Promocion Cientıfica y Tec-nologica (ANPyCT) and Universidad Nacional del ChacoAustral (UNCAus) They would like to thank Mara RomeroandMarıa Alicia Judis for their contribution in free fatty acidsdeterminations

References

[1] E Fernandez-Fernandez M L Vazquez-Oderiz and M ARomero-Redrıguez ldquoEffects ofmanufacturing process variableson the physicochemical and sensory characteristics of Galicianchorizo sausagerdquo Journal of the Science of Food and Agriculturevol 82 no 3 pp 273ndash279 2002

[2] A M Baka E J Papavergou T Pragalaki J G Bloukas and PKotzekidou ldquoEffect of selected autochthonous starter cultureson processing and quality characteristics of Greek fermentedsausagesrdquo LWTmdashFood Science and Technology vol 44 no 1 pp54ndash61 2011

[3] M G Bonomo A Ricciardi and G Salzano ldquoInfluence ofautochthonous starter cultures on microbial dynamics andchemical-physical features of traditional fermented sausages ofBasilicata regionrdquo World Journal of Microbiology and Biotech-nology vol 27 no 1 pp 137ndash146 2011

[4] R Capita S Llorente-Marigomez M Prieto and C Alonso-Calleja ldquoMicrobiological profiles pH and titratable acidity ofchorizo and salchichon (two Spanish dry fermented sausages)manufactured with ostrich deer or pork meatrdquo Journal of FoodProtection vol 69 no 5 pp 1183ndash1189 2006

[5] F Leroy J Verluyten and L de Vuyst ldquoFunctional meat startercultures for improved sausage fermentationrdquo International Jour-nal of Food Microbiology vol 106 no 3 pp 270ndash285 2006

[6] J Fernandez-Lopez E Sendra E Sayas-Barbera C Navarroand J A Perez-Alvarez ldquoPhysico-chemical andmicrobiologicalprofiles of lsquosalchichonrsquo (Spanish dry-fermented sausage)enriched with orange fiberrdquo Meat Science vol 80 no 2 pp410ndash417 2008

[7] M Bedia L Mendez and S Banon ldquoEvaluation of differentstarter cultures (Staphylococci plus Lactic Acid Bacteria) insemi-ripened Salami stuffed in swine gutrdquoMeat Science vol 87no 4 pp 381ndash386 2011

[8] M Hugas and JMMonfort ldquoBacterial starter cultures formeatfermentationrdquo Food Chemistry vol 59 no 4 pp 547ndash554 1997

[9] F-K Lucke ldquoUtilization of microbes to process and preservemeatrdquoMeat Science vol 56 no 2 pp 105ndash115 2000

[10] H J Buckenhuskes ldquoSelection criteria for lactic acid bacteria tobe used as starter cultures for various food commoditiesrdquo FEMSMicrobiology Reviews vol 12 no 1ndash3 pp 253ndash271 1993

[11] F Toldra ldquoMeat fermentationrdquo in Handbook of Food ScienceTechnology and Engineering Y H Hui Ed vol 4 CRC Press2006

[12] G Tabanelli F Coloretti C Chiavari L Grazia R Lanciottiand F Gardini ldquoEffects of starter cultures and fermentationclimate on the properties of two types of typical Italian dryfermented sausages produced under industrial conditionsrdquoFood Control vol 26 no 2 pp 416ndash426 2012

[13] N Z P PrpichM P CastroM E Cayre O A Garro andGMVignolo ldquoAutochthonous starter culture selection to keep tra-ditions in the manufacture of dry sausages aliverdquo Annals ofMicrobiology 2014

[14] ICMSF Microorganisms in Foods 1 Their Significance andMethods of Enumeration University of Toronto Press TorontoCanada 2nd edition 1987

[15] International Organization for Standardization ldquoMicrobiologyof food and animal feeding stuffsmdashhorizontal method forthe enumeration of sulfite-reducing bacteria growing underanaerobic conditionsrdquo ISO Standard 152132003 InternationalOrganization for Standardization Geneva Switzerland 2003

[16] USDAFSIS Detection Isolation and Identification ofEscherichia coli O157H7NM from Meat Products RevisionMLG 505 2010

[17] US Food and Drug Administration (FDA) ldquoSalmonellardquo inBacteriological Analytical Manual chapter 5 US Food andDrug Administration (FDA) 2007

[18] International Organization for Standardization ldquoMicrobiologyof food and animal feeding stuffsmdashhorizontal method for thedetection and enumeration of Listeria monocytogenesmdashpart 1detection method Amedment 1 modification of the isolationmedia and the haemolysis test and inclusion of precision datardquoISO Standard 11290-11996A12004 International Organizationfor Standardization Geneva Switzerland 2004

[19] Association of Official Analytical Chemists Official Methods ofAnalysis Association of Official Analytical Chemists Washing-ton DC USA 14th edition 1995


[21] R Sinnhuber and T Yu ldquoThe 2-thiobarbituric acid reaction Anobjective measure of the oxidative determinating occurring infats and oilsrdquo Journal of Japan Society of Fish Science vol 26 no5 pp 259ndash267 1977

[22] E G Bligh and W J Dyer ldquoA rapid method of total lipidextraction and purificationrdquo Canadian Journal of Biochemistryand Physiology vol 37 no 8 pp 911ndash917 1959

[23] L Hartman and R Lago ldquoA rapid preparation of fatty acidmethyl from lipidsrdquo Laboratory Practice vol 22 no 3 pp 475ndash476 1973

[24] R Talon S Leroy I Lebert et al ldquoSafety improvement andpreservation of typical sensory qualities of traditional dryfermented sausages using autochthonous starter culturesrdquo Inter-national Journal of Food Microbiology vol 126 no 1-2 pp 227ndash234 2008

[25] J Samelis J Metaxopoulos M Vlassi and A Pappa ldquoStabilityand safety of traditional Greek salamimdasha microbiological ecol-ogy studyrdquo International Journal of Food Microbiology vol 44no 1-2 pp 69ndash82 1998

[26] N Zdolec M Hadziosmanovic L Kozacinski et al ldquoMicrobialand physicochemical succession in fermented sausages pro-duced with bacteriocinogenic culture of Lactobacillus sakei andsemi-purified bacteriocin mesenterocin Yrdquo Meat Science vol80 no 2 pp 480ndash487 2008

[27] A Casaburi M-C Aristoy S Cavella et al ldquoBiochemical andsensory characteristics of traditional fermented sausages ofVallo di Diano (Southern Italy) as affected by the use of starterculturesrdquoMeat Science vol 76 no 2 pp 295ndash307 2007

[28] B Gonzalez andVDıez ldquoThe effect of nitrite and starter cultureon microbiological quality of lsquochorizorsquomdasha Spanish dry curedsausagerdquoMeat Science vol 60 no 3 pp 295ndash298 2002


[29] A Vernam and J Sutherland Meat and Meat Products Chap-man amp Hall London UK 1995

[30] B Marchesini A Bruttin N Romailler R S Moreton C Stuc-chi and T Sozzi ldquoMicrobiological events during commercialmeat fermentationsrdquo Journal of Applied Bacteriology vol 73 no3 pp 203ndash209 1992

[31] L O Sunesen and L H Stahnke ldquoMould starter cultures for drysausagesmdashselection application and effectsrdquoMeat Science vol65 no 3 pp 935ndash948 2003

[32] I Astiasaran R Villanueva and J Bello ldquoAnalysis of proteol-ysis and protein insolubility during the manufacture of somevarieties of dry sausagerdquoMeat Science vol 28 no 2 pp 111ndash1171990

[33] N Palavecino M Castro E Cayre and O Garro ldquoCaracter-izacion fisicoquımica y microbiologica de productos carnicosfermentados elaborados en la provincia del Chacordquo in I Jor-nadas Regionales de Investigacion en Ingenierıa UniversidadTecnologica Nacional Chaco Argentina 2010

[34] M C Romero A M Romero M M Doval and M A JudisldquoNutritional value and fatty acid composition of some tradi-tional Argentineanmeat sausagesrdquo Food Science and Technology(Campinas) vol 33 no 1 pp 161ndash166 2013

[35] R Casquete M J Benito A Martın S Ruiz-Moyano EAranda and M G Cordoba ldquoMicrobiological quality ofsalchichon and chorizo traditional Iberian dry-fermentedsausages from two different industries inoculated withautochthonous starter culturesrdquo Food Control vol 24 no 1-2pp 191ndash198 2012

[36] V Ferreira J Barbosa S Vendeiro et al ldquoChemical andmicrobiological characterization of alheira a typical Portuguesefermented sausage with particular reference to factors relatingto food safetyrdquoMeat Science vol 73 no 4 pp 570ndash575 2006

[37] M C Garcıa Fontan JM Lorenzo SMartınez I Franco and JCarballo ldquoMicrobiological characteristics of Botillo a Spanishtraditional pork sausagerdquo LWTmdashFood Science and Technologyvol 40 no 9 pp 1610ndash1622 2007

[38] H Ockerman Quality Control of Post-Mortem Muscle andTissue Columbus Ohio USA Department of Animal ScienceOhio State University 1976

[39] A G M Scannell C Hill R P Ross G Schwarz and E KArendt ldquoEffect of nitrite on a bacteriocinogenic Lactococcuslactis transconjugant in fermented sausagerdquo European FoodResearch and Technology vol 213 no 1 pp 48ndash52 2001

[40] D Ansorena and I Astiasaran ldquoEffect of storage and packagingon fatty acid composition and oxidation in dry fermentedsausages made with added olive oil and antioxidantsrdquo MeatScience vol 67 no 2 pp 237ndash244 2004

[41] M C D Fernandez and J M Z Rodrıguez ldquoLipolytic andoxidative changes in lsquoChorizorsquo during ripeningrdquo Meat Sciencevol 29 no 2 pp 99ndash107 1991

[42] J Goslashtterup K Olsen S Knoslashchel K Tjener L H Stahnkeand J K S Moslashller ldquoColour formation in fermented sausagesby meat-associated staphylococci with different nitrite- andnitrate-reductase activitiesrdquo Meat Science vol 78 no 4 pp492ndash501 2008

[43] S Dellaglio E Casiraghi and C Pompei ldquoChemical physicaland sensory attributes for the characterization of an Italian dry-cured sausagerdquoMeat Science vol 42 no 1 pp 25ndash35 1996

[44] E Sayas A Perez J Fernandez andMOnate ldquoCaracterizacionfısica y fısicoquımica de la Longaniza Imperial de LorcardquoAlimentaria vol 7 pp 27ndash32 1998

[45] B Rubio B Martınez M D Garcıa-Cachan J Rovira and IJaime ldquoEffect of the packaging method and the storage time onlipid oxidation and colour stability on dry fermented sausagesalchichon manufactured with raw material with a high level ofmono andpolyunsaturated fatty acidsrdquoMeat Science vol 80 no4 pp 1182ndash1187 2008

[46] F Toldra ldquoThe enzymology of dry-curing meat productsrdquo inNew Technologies for Meat and Meat Products F J SmuldersF Todra J Flores and M Prieto Eds EcceamstAudetNijmegen The Netherlands 1992

[47] J L Navarro M I Nadal L Izquierdo and J Flores ldquoLipolysisin dry cured sausages as affected by processing conditionsrdquoMeat Science vol 45 no 2 pp 161ndash168 1997

[48] K Molly D Demeyer G Johansson M Raemaekers M Ghis-telinck and I Geenen ldquoThe importance of meat enzymes inripening andflavour generation in dry fermented sausages Firstresults of a European projectrdquo Food Chemistry vol 59 no 4 pp539ndash545 1997

[49] C Coutron-Gambotti and G Gandemer ldquoLipolysis and oxi-dation in subcutaneous adipose tissue during dry-cured hamprocessingrdquo Food Chemistry vol 64 no 1 pp 95ndash101 1999

[50] J A Ordonez E M Hierro J M Bruna and L de La HozldquoChanges in the components of dry-fermented sausages duringripeningrdquoCritical Reviews in Food Science andNutrition vol 39no 4 pp 329ndash367 1999

[51] R Talon S Leroy-Setrin and S Fadda ldquoBacterial startersinvolved in the quality of fermentedmeat productsrdquo in ResearchAdvances in the Quality of Meat and Meat Products F ToldraEd pp 175ndash191 Research Signpost Trivandrum India 2002

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


a crucial role in this fermentation that is acidification whilethey can also have proteolytic and lipolytic activities Thepositive technological features of this acidification include (i)inhibition of spoilage and pathogenic microflora (ii) quickdrying and enhancement of texture due to denaturation andcoagulation of proteins (iii) activation of muscle proteasesand (iv) formation of nitric oxide and nitrosomyoglobinwhich contribute to redness [9] However an excessive acidi-fication can negatively affect the taste or smell of the productand lead to color failures as a consequence of CNC inhibition[10] The most important factors determining characteristicsand quality of meat fermented products are the selectionof starter cultures and the environmental conditions duringfermentation and ripening [11 12] Quality of the productswill be the result of the interaction between these two factorswhich generates a beneficial or a disadvantageous setting forcellular metabolism

Indigenous microorganisms from meat fermentation arewell adapted to fermentation and ripening conditions and areable to inhibit spontaneous growth of undesirable microflora[8] Selected autochthonous starter cultures (SAS) can con-tribute to the manufacture of artisanal local products withparticular features In previous studies two strains of L sakeione strain of S vitulinus and another of S xylosus wereisolated from regional meat fermented products [13] Owingto their safety and technological features these strains havepotential application as starter cultures In order to evaluatetheir performance in situ and consequently the feasibilityof their use these strains were tested as starter cultures in alocal small-scale facility which manufactures dry fermentedsausages Two different mixed starter cultures were designedMicrobiological and physicochemical characteristics of theproducts had beenmonitored throughout the manufacturingprocess while sensory attributes of the final products hadbeen evaluated by a trained panel

2 Materials and Methods

21 Autochthonous Starter Cultures Strains used in this studywere selected in previous studies taking into considerationtheir technological and safety features [13] they are reportedin Table 1 Association of the different strains to design eachstarter culture was determined by means of compatibilitytests Microorganisms are deposited in the Strain Repositoryof the Laboratorio de Microbiologıa de Alimentos (Universi-dad Nacional del Chaco Austral Argentina) Lactobacilli arekept in MRS broth (de Man Rogosa Sharpe) supplementedwith 20 vv glycerol while staphylococci are kept in soytrypticase + 06wv yeast extract (TSBYE) added with20 vv glycerol both bacterial species aremaintained frozenat minus18∘C

Two mixed starter cultures were designed namely SAS-1 comprised of L sakei 487 and S vitulinus C2 and SAS-2comprised of L sakei 442 and S xylosus C8 Lactic bac-teria were propagated in MRS broth and CNC strains inTSBYE they were incubated at 30∘C for 12 h Cells were thenharvested by centrifugation (4500 rpm 20min) washedtwice with saline peptone water (085 gNaCl100mL) and

Table 1 Indigenous bacterial strains used as starter cultures in thisstudy

Strain Technological activity



Staphylococcus vitulinus C2 Proteolytic and nitrate reductaseStaphylococcus xylosus C8 Lipolytic and nitrate reductase

resuspended in this solution Cellular density was monitoredby plate count (MRS agar for LAB and Mannitol salt agar forCNC)

22 Dry Fermented SausagesManufacture and Sampling Dryfermented sausages used in this work were those currentlymanufactured by a local small-scale facility Traditionalrecipes dictate the composition of these meat products perkilogram that consists of 450 g beef 177 g pork meat 300 gbacon 34 g sodium chloride 15 g sugar 10 g powder milk07 g potassium nitrate 3 g sodium glutamate 5 g choppedgarlic 2 g ground red pepper and 3 g ground black pepperAll the ingredients were thoroughlymixed together andmeatdough was divided into two batches one batch was addedwith the SAS and the other was kept as a control systemAfterwards meat dough was packed and shaped into naturalcasings (sheep gut) SASwere added once themeat dough hadbeen finished being completely homogenized before casingswere stuffed Cultures SAS-1 and SAS-2were added separatelyto the meat dough in two different instances Final bacterialconcentration (LAB + CNC) reached a value of sim106 colonyforming units per gram (cfug) After inoculation and stuff-ing the unfermented products were placed in a ripeningchamber for 15 days During the first seven daysmdashferment-ation periodmdashtemperature was kept at 18ndash22∘C and relativehumidity (RH) oscillated to 90ndash95 The next 8 daysmdashripening periodmdashtemperature was kept at 15∘C andRH rangewas 80ndash85 From each batch samples were withdrawn toperform physicochemical and microbiological analyses atinitial time 2 4 7 and 15 days

23 Microbiological Analyses Ten grams of each sample wasaseptically taken and was transferred to a stomacher bag with90mL of sterile peptone water to be homogenized After2min in the stomacher (Stomacher 400 Circulator SewardUK) appropriate dilutions of the homogenate were done inorder to inoculate several growth media to determine viablecounts of LAB in MRS agar (30∘C 72 h) Micrococcaceaein AMS (30∘C 72 h) yeasts and molds in ChloramphenicolSabouraud Agar (25∘C 5 days) Enterobacteriaceae in Glu-cose Red Violet Bile Agar (37∘C 24 h) Staphylococcus aureusin Baird Parker Agar with egg yolk emulsion and potassiumtellurite (37∘C 48 h) Besides final productswere also assayedfor the following Escherichia coli count (MPNg) follow-ing the ICMSF method [14] sulphite-reducing anaerobes(cfug) according to ISO 152132003 [15] absence of E coli

















1

2

3

4

5

6

7

8

9



log 1

0(c

fug

)

(a)

1

2

3

4

5

6

7

8

9


log 1

0(c

fug

)

(b)

0

1

2

3

4


log 1

0(c

fug

)

(c)

0

1

2

3

4


log 1

0(c

fug

)

(d)






4

5

6

7

0 5 10 15

pH


(a)

20

30

40

50

60

70

0 5 10 15

Moi

sture

()


(b)

75

80

85

90

95


aw

()


119908

















System 119871

lowast

119886

lowast

119887

lowast












01234567

Easy peeling

Cohesiveness

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast



capabilitylowast

(a)

01234567


Cohesiveness

Color intensity

Coloruniformity

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

(b)



4 Conclusions






Acknowledgments


References




























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

















1

2

3

4

5

6

7

8

9



log 1

0(c

fug

)

(a)

1

2

3

4

5

6

7

8

9


log 1

0(c

fug

)

(b)

0

1

2

3

4


log 1

0(c

fug

)

(c)

0

1

2

3

4


log 1

0(c

fug

)

(d)






4

5

6

7

0 5 10 15

pH


(a)

20

30

40

50

60

70

0 5 10 15

Moi

sture

()


(b)

75

80

85

90

95


aw

()


119908

















System 119871

lowast

119886

lowast

119887

lowast












01234567

Easy peeling

Cohesiveness

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast



capabilitylowast

(a)

01234567


Cohesiveness

Color intensity

Coloruniformity

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

(b)



4 Conclusions






Acknowledgments


References




























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


4

5

6

7

0 5 10 15

pH


(a)

20

30

40

50

60

70

0 5 10 15

Moi

sture

()


(b)

75

80

85

90

95


aw

()


119908

















System 119871

lowast

119886

lowast

119887

lowast












01234567

Easy peeling

Cohesiveness

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast



capabilitylowast

(a)

01234567


Cohesiveness

Color intensity

Coloruniformity

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

(b)



4 Conclusions






Acknowledgments


References




























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology











System 119871

lowast

119886

lowast

119887

lowast












01234567

Easy peeling

Cohesiveness

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast



capabilitylowast

(a)

01234567


Cohesiveness

Color intensity

Coloruniformity

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

(b)



4 Conclusions






Acknowledgments


References




























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








01234567

Easy peeling

Cohesiveness

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast



capabilitylowast

(a)

01234567


Cohesiveness

Color intensity

Coloruniformity

Aroma intensity


Salty

Acid

Sour

Hardening

Chewing

Firmnesslowast

(b)



4 Conclusions






Acknowledgments


References




























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


Acknowledgments


References




























































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology
































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Research Article Indigenous Starter Cultures to Improve ...

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