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Small Ruminant Research 101 (2011) 17 32
Contents lists available at SciVerse ScienceDirect
Small Ruminant Research
jou rn al h om epa ge: www. elsev ier .com/ locate /smal l rumres
Microb adi
EvanthiaLaboratory of F , FacultyThessaloniki, G
a r t i c l
Article history:Available onlin
Keywords:Greek traditioMicrooraBiochemical aNSLAB
centurd gift. em we
g to thees, harg is conheese
of the usuall
1. Introdu
Cheese making in Greece has a centuries old tradition.Homer in 8th century BC in his Odyssey describes wellthe shepherd and cheese maker Polyphemus of the 12thcentury BC,lus, 1st centthat Aristeulearned thenymphs, wteach the Gcheese as a fsidered the that HomerFeta and it ithe ancienttypes evolvarea, almosin cheese m
Lately, thtional techn
This papeRuminants, Gu
CorresponE-mail add
ealthiestablished as a recent way to understand the quality ofa product (Rubino et al., 1999). There is also the CouncilRegulation (EEC) no. 2087/92 (1992), which attempts tounify different concepts of typicity and lays down rules on
0921-4488/$ doi:10.1016/j. as well as his cheeses. Diodorus (Diodorus Sicu-ury B.C), the Greek historian from Sicily, wrotes, son of Apollo and grandson of Zeus, who had
art of cheese making from his nannies, theas sent by the Gods at Olympus mountain toreeks how to make cheese. Given the value ofood, it is not surprising that ancient Greeks con-cheese as a divine invention and gift. The cheese
described seems possible to be the ancestor ofs the main cheese manufactured in Greece from
times till today. Meanwhile, various cheeseed through the centuries, so that nowadays eacht every island has its individual unique traditionanufacture.e EU supports the local products and the tradi-ologies. Typical foods are considered as safer
r is part of the special issue entitled Products from Smallest Edited by A. Govaris and G. Moatsou.
ding author. Tel.: +30 2310991727; fax: +30 2310991674.ress: ganet@agro.auth.gr (E. Litopoulou-Tzanetaki).
the protection of agricultural products intended for humanconsumption. The PDO/PGI labels guarantee specic sen-sory properties and imply that particular characteristics arepresent in the product.
Numerous traditional cheeses are made throughoutGreece today. Some of them are in fact types of the samecheese variety, have somewhat different steps in tech-nology or possibly the same technology, but known withdifferent local names. Twenty of them were recognizedas PDO (Protected Designation of Origin) cheeses andthere are others, waiting for their recognition. As a total,Greek traditional cheeses could be grouped, according totheir technology of manufacture, as: cheeses in brine; softcheeses; semi-hard cheeses; hard cheeses; whey cheeses.
2. Cheeses in brine (Pickled cheeses)
2.1. Batzos cheese, PDO
Batzos is a low-fat, semi-hard, white brined cheese witha sourish and lightly piquant taste and a large number of
see front matter 2011 Elsevier B.V. All rights reserved.smallrumres.2011.09.022iological characteristics of Greek tr
Litopoulou-Tzanetaki , Nikolaos Tzanetakisood Microbiology and Hygiene, Department of Food Science and Technologyreece
e i n f o
e 2 October 2011
nal cheeses
ctivities
a b s t r a c t
Cheesemaking in Greece has acheese as a divine invention anGreece today and twenty of thcheeses are grouped, accordinsoft cheeses, semi-hard cheesmicroora throughout ripenintheir biochemical activities to ctics. Microorganisms indicativedecline during ripening and are
ction and htional cheeses
of Agriculture, Aristotle University of Thessaloniki, 54124
ies-old tradition and ancient Greeks considered theNumerous traditional cheeses are made throughoutre recognized as PDO. As a total, Greek traditionalir technology of manufacture, as: cheeses in brine,d cheeses and whey cheeses. Their predominantsisted of NSLAB that proliferate and contribute byripening and the development of avor characteris-ir hygienic quality, Enterobacteriaceae and coliforms,y found at negligible levels in the mature products.
2011 Elsevier B.V. All rights reserved.
er by the consumer and typicality has been
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18 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
holes in the body. It is made from either caprine milk, orovine milk or mixtures of both milks. The cheese is madeat home by the traditional method as well as at cream-eries of Western Macedonia and the nearby Thessaly, usingmodications of the traditional method and technologicalinnovations.
The milk is not heat-treated and coagulation takes placewithin 50 min at 2830 C. When the milk just begin to gel,the manufacturer hits the milk with a thick wooden stickabout 150200 times; the milk is then left to curdle for3540 min before being hit again for 300350 times. Duringthese hitting stages, a large proportion of fat is transferredto the whey. The curd is then left to settle, but some cheesemakers would apply mild heat before placing the curd, cutin appropriate pieces, in cheese cloths and hanging themto drain and ripen for 24 h. The next day, the ripened curdis cut into slices, salted with coarse salt, placed in tinsand covered with brine (Anifantakis, 1991; Zygouris, 1956).Cheese is either consumed fresh or stored in cool rooms.
The microbial counts in the curds of Batzos cheese fromovine milk in summer (Fig. 2) were higher than in curdsof spring cheeses (Fig. 1) for all microbial groups exceptstaphylococci (Nikolaou et al., 2002). Lactic acid bacte-ria (LAB), Enterobacteriaceae and coliforms are the majorcomponents of the microora during ripening; LAB grownon M17 agar was the most abundant microbial group inthe curd and cheese during storage for one month at 4 C.Enterobacteriaceae and coliforms, declined faster in cheesesmade in the summer. This was possibly related to the lowerpH values of cheeses made in the summer and the highernumber of LAB present in curds of summer cheeses.
Among LAB lactobacilli predominated in cheeses madein summer;
Table 1Species of lactic acid bacteria isolated from ripened curd of Batzos cheesefrom raw ovine milk, made in spring and summer. Isolates from twobatches of cheese in each season.
% of isolates from cheese made in
Species Spring Summer
LactococciLc. lactis subsp. lactis 22.22
LactobacilliLb. paracasei subsp. paracasei 32.98Lb. paraplantarum 14.82 30.85Lb. pentosus 7.41
EnterococciE. faecalis 7.41 13.83E. faecium 40.74 21.28E. durans 3.70 E. pseudoavium 1.06Pediococcus spp. 3.70
Total 100.00 100.00
bacteria (Table 1). Lactobacillus paracasei subsp. paracaseiand Lb. paraplantarum were isolated frequently from theripened curds of Batzos cheese in summer, while Enterococ-cus faecium was the most frequently isolated LAB speciesin spring. Lactococcus lactis subsp. lactis was found only inthe curds of spring cheeses.
Hydrolysis of both s- and -casein (CN) proceededmore rapidly in spring than in summer; s-CN is alsodegraded faster than -CN. Lipolysis products increasedgradually with storage and accumulated at higher levelsin cheese made in the summer.
For the cheese from caprine milk no signicant differ-ences were observed in the counts related to the season,
ignic
0
2
4
6
8
10
12
sts Lactobac ill i Lact ococc i Enter ococci
log 1
0 cfu
/g
Fig. 1. Log cou ine milk, made in summer. Mean of two cheesemaking trials. in spring enterococci were the most abundant but s
TC Coliforms En ter obac teriace ae Staphyloc occ i Yea
nts (x SD) of microbial groups in the curd of Batzos cheese from raw ovant changes in the microbial counts throughout
Curd (afte r draining)Ripened curd
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E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 19
0
2
4
6
8
10
12
ea sts
log 1
0 cfu
/g
Curd (af ter drainin g)Ripened curd
Fig. 2. Log ovine
ripening analso observteriaceae anin cheese frmicrobial ghad a signiaceae and c
The lactnance of cein winter lacocci were spring and samong the isolation of cheeses.
The maitis, found mand summetobacilli prparaplantarisolated. Enrococcus in
In an attthe cheese strains werpasteurizedRS and PS cexcluding fpartially skmicrobial lo(R) and rawwere similaOnly when
e prolation
cheesdesirabgradall reduded. Ar rate tased thTC Colif orms Enterobac ter iace ae Staphyloc occ i Y
counts (x SD) of microbial groups in the curd of Batzos cheese from raw
d storage were recorded (Psoni et al., 2003). Ased for cheese from ovine milk, LAB, Enterobac-d coliforms are the principal microbial groupsom caprine milk. LAB, are the most abundantroup throughout storage. Ripening and storagecant inuence on the number of Enterobacteri-oliforms (Fig. 3).ation season has a clear effect on the predomi-rtain LAB in each season. Thus, in cheese made
cheescoaguotherof un
Dea smarecorfasteincrectococci and enterococci predominated. Lacto-less frequently isolated from cheeses made inummer, whereas enterococci were rarely foundpredominant LAB. However, the frequency oflactobacilli was increased in spring and summer
n lactococci species were Lc. lactis subsp lac-ore frequently in winter cheeses. In spring
r cheeses facultatively heterofermentative lac-edominated. Lactobacillus plantarum and Lb.um were the principal species of lactobacilliterococcus durans was the predominant Ente-cheese made in winter.empt to improve the microbiological quality offrom raw milk, selected Lc. lactis subsp lactise used as starter to make cheese from raw (RS),
(PS) and pasteurized standardized (PSS) milk.heese were made by the traditional method ofat from the milk. PSS cheese was made fromimed milk (Psoni et al., 2006). Surprisingly, theads of lactococci in cheeses made from raw milk
milk with starter and the traditional methodr and the same was observed with their pH.
standardized milk (25 g/L fat) was used for
Thus, coof a native sin better mstandardizenot the trad
2.2. Feta ch
Feta is a cid and sliggas holes sical openinmixtures ofproduced anot commo
The coagnet from abAfter curdlicheese clotpressing. Thand salted babout 33.5for about 1is developeLactobac illi Lactococc i En terococc i
milk, made in spring. Mean of two cheesemaking trials.
duction without hitting and stirring during, higher levels of LAB and lower pH than thee types were recorded. A higher rate of decreasele microorganisms was recorded in this cheese.tion of s-CN was in R > RS and in PS > PSS, whilection of -CN during ripening and storage wass expected, degradations of s-CN occurred at ahan -CN and proteolysis and lipolysis productsroughout ripening and storage.nclusively, cheese milk pasteurization and usetarter from the traditional cheese has resultedicrobiological quality Batzos cheese. However,d milk with low fat content must be used anditional method of excluding fat into the whey.
eese, PDO
semi-soft, white brined cheese, with mildly ran-htly acid taste, rm and smooth texture. No
hould be present, but irregular small mechan-gs are desirable. It is made from ovine or from
it with caprine milk (up to 30%). The cheese isll-over the country. However, its production isn in the islands.ulation of the milk is traditionally made by ren-omasa of lambs and kids, within 50 min to 1 h.ng, the curd is cut and transferred into eitherhs or perforated moulds for draining withoute drained curd is subsequently cut into blocksy coarse salt on cheese-tables, until it contains% salt. The cheese remains on the cheese-table5 days. During this period a surface microorad which is considered to contribute to cheese
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20 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
Fig pening o
ripening aninto barrelsin cold room
The moand standarized to a fa(72 C, 15 sneting. Lactbulgaricus) ing to Greekripens for a
The pred4d, when tposed of lacmicrobes. Ltute the prewhile at thmicroora being the pare largely either not dand Hatzikat 4 days ahansenii, wS. cerevisiaeleu- and vaaminopeptiactivity is hstrains only
alotol. 3. Log counts (x SD) of Enterobacteriaceae and coliforms throughout ri
d avor. The cheese is afterwards transferred The h
or tins with brine (78%) to continue ripenings.
dern technology follows the traditional stepsdized conditions. The milk is usually standard-t content of 5.86.0%; it is then pasteurized
) and immediately cooled at 3234 C for ren-ic starters (for example yoghurt, Lc. lactis + Lb.are added 20 min prior to renneting. Accord-
standards (Greek Food Code, 1998) the cheeset least two months.ominant surface microora of fresh cheese (athe microbial slime starts appearing) is com-tic acid bacteria (LAB), yeasts and salt toleranteuconostoc, Lc. lactis and Lb. plantarum consti-dominant LAB microora developed at day 4,e end of ripening in the warm room the LABis composed of lactobacilli, with Lb. plantarumredominant species. Aminopeptidase activitiesdue to lactobacilli, while lipolytic activities areetected or weak in these organisms (Tzanetakisamari, 1994). The predominant yeast speciesre Saccharomyces cerevisiae and Debaryomyceshile at the end of ripening in the warm room
is the predominant yeast. Yeasts also containl-aminopeptidase that is higher than the cys-dase activity. In addition, their esterase/lipaseigher than the esterase activity, and for some
lipase activity is found (Tzanetakis et al., 1996).
lococci, micStaphylococant bacteriwhile coryold cheese. do not exhstaphylococesterase/lipThus, lactobsis, yeasts pstaphylococ
The nonate during may reach of ripeningTzanetaki, lactococci iperiod and ilar behavihand, micromaximum heterofermthroughoutnant specieand Lb. paroften foundamong the and Litopof goat milk cheese made in spring and summer.
erant surface microora is composed of staphy-
rococci, enterococci and coryneform bacteria.cus saprophyticus predominates the halotoler-a content on the surface of cheese at day 4,neforms are commonly found in the 20-day-Staphylococci isolates from Feta cheese surfaceibit any aminopeptidase activity; however,ci show quite strong, in general, esterase andase enzyme activity (Tzanetakis et al., 1995a,b).acilli of the cheese surface may affect proteoly-articipate to both, proteolysis and lipolysis, andci contribute to cheese lipolysis.
starter lactic acid bacteria (NSLAB) prolifer-Feta cheese from pasteurized milk ageing andnumbers as high as 107108 cfu/g at the end
in the warm room (Tzanetakis and Litopoulou-1992; Manolopoulou et al., 2003). Mesophilicncrease signicantly their numbers during thisthen decrease; enterococci also showed a sim-or (Manolopoulou et al., 2003). On the othercocci, yeasts and coliforms seem to reach their
levels at day 4, for most cheeses. Facultativelyentative lactobacilli are an important group
cheese ripening. Lb. plantarum is the domi-s constituting the 50% of the NSLAB population,acasei subsp. paracasei and Lb. brevis are also. In addition, enterococci and pediococci arepredominant NSLAB of Feta cheese (Tzanetakisulou-Tzanetaki, 1992). Other species, such as
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E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 21
Table 2Log counts (x SD) of microbial groups during ripening of Feta cheese from raw ewes milk.
Days at 16 C Days at 4 C
1 4 30 60
TAC 9.18 0.8 8.37 0.2Enterobacter 3.04 2.6 2.49 2.2Coliforms 3.08 2.7 2.65 2.3Staphylococ 3.54 3.1 4.88 0.2LAB on MRS 8.48 0.6 8.29 0.1LAB on M17 7.68 0.2 6.80 0.1Lactobacilli 8.36 0.3 8.18 0.1Enterococci 5.68 0.7 5.53 0.2Yeasts 4.43 0.8 4.77 0.3Gram ve 4.92 0.7 2.60 2.3
Lb. paraplanquently (M
In the chdecrease thlactobacilli ripening, Enegative barococci als(Vassiliadishygienic quiforms, werNSLAB mic(Table 3). Lgroup in thdominatingenterococcivailed overcheese. ThisLb. paracase
The tradraw milk anheat treatma starter. Pthey were hmized (TS)accumulatecheese typeCN occurreamount of types. -Cawhile in TSincreasing lated in theless in TS thsibly due toincreased to
Lb. plantand peptidoThe amounlow and strate of acidienzymes casei isolatesaminopeptienzyme ac
opeptidase activity is low, except for the diagnos-bstrate Gly-Pro-pNA. This is an important feature
heese ripening since caseins have high-proline con-In addition, Lb. paracasei subsp paracasei may degradens (Bintsis et al., 2003). The amount of amino acidsulated in the milk after 24 h is low and strain depen-
(Mama et al., 2002). Moreover, the esterase activityases with increasing carbon chain length (Bintsis et al.,). Less frequently isolated during ripening of Feta,
3es in numbers and kinds of non starter lactic acid bacteria (NSLAB)
ripening of Feta cheese from raw ewes milk.
ning (days)
Species No. ofisolates
%
Lc. lactis ssp. lactis 12Lc. lactis ssp. cremoris 2Lc. rafnolactis 3 43.59Ent. durans 2Ent. faecalis 1Ent. faecium 1Ent. saccharolyticus 10.26Lb. buchneri 8Lb. brevis 19.94 1.6 8.95 0.3 iaceae 7.25 0.3 6.24 0.8
7.89 1.0 6.43 0.8 ci 6.50 0.1 5.91 1.0
agar 8.87 0.3 8.80 0.2 agar 9.01 0.3 8.65 0.3
7.94 0.4 8.04 0.0 5.98 05 6.83 0.2 6.37 1.6 5.86 2.2 7.04 0.9 6.25 0.7
tarum and Lb. pentosus may also be found fre-anolopoulou et al., 2003).eese made from raw milk, all microbial groupseir counts from one to four days, except forand enterococci (Table 2). After one month ofnterobacteriaceae, coliforms, yeasts and Gramcteria decreased their numbers rapidly. Ente-o decline, and lactobacilli increase slightly
et al., 2009). Microorganisms indicative of theality of the product, Enterobacteriaceae and col-e not detected in the 60-day-old cheese. Theroora of raw milk Feta cheese is complexactococci were the most abundant microbiale fresh cheese, with Lc. lactis subsp. lactis pre-. Lactobacilli were found more frequently than
in the fresh cheese and Lb. paraplantarum pre- the other NSLAB species in the 30-day-old
species was also equally frequently found withi subsp. paracasei in the cheese after 60 days.itional cheesemakers of Feta cheese either used no starter at all or apply a milder-than-HTSTent of milk and usually traditional yogurt as
roteolysis products increased with ageing andigher in cheese made from raw (R) than ther-
milk throughout ripening. Amino acid levelsd in the cheese increased with ripening in boths (Vassiliadis et al., 2009). Degradations of s-d at a faster rate than -CN. At 60 days theundegraded s-CN was similar in both cheesesein remained almost unattacked in R cheese,
it underwent a low degree of degradation,with ripening. The lipolysis products accumu-
cheese after 1 month of ripening were 50%
amintic sufor ctent. caseiaccumdent incre2003
Table Changduring
Ripetime
1
30 an R cheese. FFA content then decreased, pos- further degradation in R cheese, while in TS
higher than R levels (Table 4).arum isolates from Feta cheese exhibit lipolyticlytic activities (Xanthopoulos et al., 2000a,b).ts of amino acids accumulated in the milk arerain-dependent. Strains of slow and mediumcation are also found. An array of peptidolyticn be detected in Lb. paracasei subsp paraca-
from Feta such as endocellular dipeptidase,dase, endopeptidase, and carboxypeptidasetivity (Bintsis et al., 2003). Their dipeptidyl
60 W. confusa 23.08Ln. mesenteroides ssp.dextranicum
4
Ln. mesenteroides ssp.mesenteroides
5 23.08
Lc. lactis ssp. lactis Ent. faecalis 3Ent. avium 1Ent. saccharolyticus Ent. hirae 8.16Lb. paraplanarum 30Lb. paracasei ssp. paracasei 9
Lb. plantarum 1Lb. rhamnosus 3Lb. buchneri 1Lb. brevis W. confusa 1 89.98Ln. mesenteroides ssp.mesenteroides
2.04
Lb. paraplanarum 18Lb. paracasei ssp. paracasei 18Lb. paracasei ssp. tolerans 9Lb. plantarum Lb. buchneri 2Lb. brevis 100TOTAL 135
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22 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
Table 4Values (x SD)a of pH, moisture %, NaCl-in-moisture %, proteolysis (o-PA)b, degradation of caseins and lipolysis (acid degree value ADV) during ripeningof Feta cheese made from raw (R) and/or thermized (TS) ewes milk.
Days at 16 C Days at 4 C
pH 0.2 0.0
Moisture% 2.2 2.4
NaCl-in-moi 2.6 0.3
Proteolysis (
% degradatios-CN
-CN
Lipolysis (AD
a Mean of thb In ppm of
c,dValues of ea
heterofermesterase, aacids (Vafotoc strains mthe caseinsTzanetaki, 1
Pediococacid slowlywhen growThese orgaand dipept(Vafopoulostrains alsonicant intactivity on casein. Qupentosaceus
Enterocoand some (AmbadoyiLow peptidand their ecocci seemmolecular wThe main voacetaldehyd
Enterocolow pH (3.0ited antagosporogenes had interescould be usfrom the suinteresting assimilated
It may bLAB microtribute to pby virtue of
nyl co abilite inicrob
istics tigatiohen tphilicdjunc
to mere a
the ads of rited hides, pitopoubservto maknhanceentrati). The Cheese type 1 4
R 4.96c 0.1 4.81cTS 4.88c 0.1 5.0cR 55.03c 1.4 51.1cTS 55.59c 0.7 53.8c
sture% R 1.956c
TS 1.582c
o-PA) R 2412.21c 245.93 TS 1997.96c 272.72
n of caseinsR TS R TS
V) R TS
ree cheesemaking trials.l-glycine equivalents.ch raw of the same cheese differ (P < 0.05).
entative lactobacilli and leuconostocs, producend degrade preferentially short-chain fattypoulou-Mastrogiannaki et al., 1996). Leuconos-
ay exhibit peptidase activities and hydrolyze (Vafopoulou-Mastrogiannaki and Litopoulou-996).cus pentosaceus isolates from Feta produce
and often form diacetyl and acetaldehyden in milk (Litopoulou-Tzanetaki et al., 1989).nisms also have dipeptidyl aminopeptidaseidase activity for peptides containing prolineu-Mastrogiannaki et al., 1994). P. pentosaceus
exhibit carboxypeptidase activities with sig-er-strain differences. Their overall proteolytic-casein is considerably higher than on s1-ite low esterase activities are found in P.
compared to other organisms.cci isolates from Feta are poor acidiersof them exhibit low proteolytic activity
annis et al., 2005; Sarantinopoulos et al., 2001).ase activities are also exhibited by enterococci
carboifying
Ththe macterinves
Wmesoand ausedues wand stagemulapeptivor (Lalso oture via econc2002sterolytic system is rather complex; entero- to preferentially degrade substrates with loweight fatty acids (Sarantinopoulos et al., 2001).latile compounds produced by enterococci aree, ethanol and acetoin.cci isolates from cheese were able to withstand) and high bile (1%) concentrations and exhib-nistic activities towards enterococci, LAB and Cl.(Ambadoyiannis et al., 2005). Selected isolatesting probiotic and technological properties anded as adjuncts to make cheese. Isolates of yeastsrface of Feta cheese were also found to exhibitprobiotic properties (Psomas et al., 2001) and
cholesterol into the cells (Psomas et al., 2003).e concluded, therefore that, the predominantora during ripening of Feta cheese may con-roteolysis and avor development of the cheese
an active proteolytic system, the ability to form
acetate, aceamounts de
When P.for Feta chelower levelthe fresh chtrol cheeselevels of acwere also siP. pentosaceration time
2.3. Toulou
Touloumskin bags (Tmodicatiofrom raw o30 60
4.69c 0.5 5.06c 0.64.83c 0.3 5.06c 1.0
56.13c 6.7 60.99c 9.758.63c 5.9 62.0c 7.46.098d 1.7 5.355cd 0.56.724d 0.3 4.582cd 0.3
5929.77d 257.87 7383.21d 659.235241.73d 948.70 5087.02d 1125.64
57.07c 13.67 66.7c 19.0139.4c 7.68 65.2c 10.380.44c 0.24 0.44c 0.241.73c 0.59 10.17d 2.66
0.559c 0.008 0.288d 0.0940.244c 0.132 0.349c 0.08
mpounds, their lipolytic activity and their acid-ies.uence of the various starter combinations oniological, physicochemical and sensory char-of Feta cheese has been studied in severalns.hermophilic starter cultures, combinations of
LAB (Lc. lactis subsp lactis, Lactobacillus casei)t cultures (E. durans, Leuconostoc cremoris) wereake Feta cheese, signicantly lower pH val-ttained in the cheese with the mesophilic LABjuncts. At the same time, during the laterpening the adjunct E. durans population accu-gher concentrations of amino acids and smallroducing cheese with signicantly better a-lou-Tzanetaki et al., 1993). Similar effects were
ed by the addition of E. faecium as an adjunct cul-e Feta cheese. The cheese avor was improvedd degradation of s- and -CN and increasingons of free amino acids (Sarantinopoulos et al.,main volatile compounds formed were ethanol,
taldehyde, acetoin and diacetyl, with highesttermined for ethanol, followed by acetate.
pentosaceus was included in the starter cultureese production, the cheese pH was reduced tos and the amount of soluble nitrogen, even ineese, was signicantly higher than in the con-
(Vafopoulou-Mastrogiannaki et al., 1990). Theetaldehyde in cheese made with P. pentosaceusgnicantly higher and the avor was improved.us accelerated cheese ripening and the matu-
of Feta cheese by 1 month.
missio cheese (Touloumotyri)
issio is a cheese that ripens and is preserved inouloumia). It is in fact Feta cheese with somens in technology. The cheese is manufacturedvine or caprine milk or mixtures of them. The
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E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 23
milk is curdled as for Feta. After salting and ripening, thecheese is cut in small pieces and put into skin bags. Theproduction of this cheese has been dramatically reducednowadays, mainly due to its preservation.
The pH Feta (4.65microora (be found at2.236.07, that the com(brine concundesirableples were n
2.4. Kalatha
This chenique simildrained int
2.5. Sfela (F
Sfela is ponnesus, poften called3638 C. Ttures of ovibarrels, likecurd is draiwhere it is shape. The cit is dry-sal
2.6. Teleme
Teleme cheese, maTeleme seeIt is manufmainly.
The techof that useding. The moexpel the ware immed20 h) to allo
LAB growmilk is simithe predomtococci are 75 d, while group untilTzanetaki, 1leuconostocFeta, in resp(Mama et a1989; VaTzanetaki, 11996). Whelactis and Lmoris and E
was lower than in cheese made with thermophilic startercultures (Tzanetakis et al., 1995a,b). In addition, the aminoacid levels in these cheeses were higher and the cheeseavor was improved. P. pentosaceus is also benecial as
junct nce, the wase proerationferent
of misis (Pare) anoris) stst degrescendows monclu
e thermaking
ted in
ft che
nevat
evatoprine m
pleasmilk. eries
rth-weaditioacedo. Theye takiurd wan late
villagee.e micrmade(Hatziram-nheese Howevecreasourabf LAB
itions r winteriaceabial g
in Mne in cry.ctococt 4 C a0-dayevel insaltingof the cheese is sometimes higher than in5.40), lactobacilli constitute the predominat107 cfu/g) and enterococci and yeasts may also
high levels (range of log10 cfu/g 5.008.27 andrespectively) (Tzanetakis, 1990). It also seems,
bined effect of low pH and high salt contententration 5.567.35%) regulate the survival of
microorganisms. Thus, 35% of the retail sam-egative for coliforms.
ki of Limnos, PDO
ese type is made at Limnos island with a tech-ar to Feta, with the exception that the curd iso round baskets, where it is also surface salted.
eta of re), PDO
a semi-hard cheese produced in South Pelo-articularly in Messinia province. The cheese is
Feta of re because the curd is cooked athe cheese is made from raw ovine milk, mix-ne and caprine milk, and it nowadays ripens in
Feta. The milk is curdled like Feta. The cookedned in cheese cloths and it is then put on tables,inverted regularly, in order to obtain a roundheese is then cut in pieces, the size of Feta, andted (Anifantakis, 1991).
is a soft and pleasant in taste white-brinedde from all milk types or mixtures of them.ms to have originated from northern Romania.actured all over the country, in the mainland,
nology of manufacture of Teleme cheese differs for Feta in the procedure of draining and salt-ulded curd of Teleme, is subjected to pressure tohey. After draining the blocks of Teleme, theyiately immersed into brine (usually 18% NaCl,w penetration of salt (Anifantakis, 1991).th and evolution in Teleme cheese from ewe
lar to that in Feta, with Lb. plantarum becominginant species with the progress of ripening. Lac-present in curd and cheese until the age of up toleuconostocs constitute an important microbial
30 d of ripening (Tzanetakis and Litopoulou-992). Isolates of P. pentosaceus, lactobacilli ands from Teleme are also similar to isolates fromect of their proteolytic and lipolytic activitiesl., 2002; Tzanetakis and Litopoulou-Tzanetaki,fopoulou-Mastrogiannaki and Litopoulou-996; Vafopoulou-Mastrogiannaki et al., 1994,n mesophilic starter cultures (Lc. lactis subspb. casei) were used concurrently with Ln. cre-. durans to make Teleme the pH of the cheese
an adpresecheescheesaccelof diftypesteolycultucremloweof a dand calso cor ththe mresul
3. So
3.1. A
Anor cahas agoat creamin no
Trern Msheepbeforthe creturtheirchees
Thmilk high and Gthe cson. are dunfavtion ocondand/oobactmicromadedecliJanua
Laage aand 6low lafter starter culture (Tzanetakis et al., 1991). In itse growth of LAB was enhanced, the pH of the
signicantly lower than in the control and theteolysis was accelerated. Thus, a concomitant
of ripening by 1 month was noticed. The use starter cultures to make cheese from differentlk was found to affect the rate of cheese pro-ppa et al., 2006). The thermophilic (yoghurtd mesophilic (Lc. lactis subps lactis and subsparter resulted in cheeses with the highest andee of proteolysis, respectively. Proteolysis wasing order in cheeses made from sheeps, goatsilk on the basis of nitrogenous fractions. It was
ded that the mixed (thermophilicmesophilic)ophilic cultures might be more appropriate in
of Teleme cheese from all milk types, as theycheeses of higher levels of proteolysis.
eses
o, PDO
is a spread type cheese made from raw ovineilk or mixtures of both. The traditional cheese
ant taste, especially the product made fromThe cheese is produced at home as well as at
in the mountain region of Western Macedoniast Greece and the nearby Thessaly.
nally, Anevato cheese was produced in West-nia by shepherds with large ocks of goats and
renneted milk obtained in the morning justng the cattle out for feeding. During the day,s raised and was ready to be drained on theirin the afternoon. The shepherds used to visits once a week to see their families and sell their
obial counts of the traditional cheese from goat throughout the whole lactation season werekamari et al., 1999). Lactic acid bacteria (Fig. 4)egative organisms are the major components ofmicroora throughout the entire lactation sea-er, Enterobacteriaceae and coliforms numbersed even during curd draining, due possibly tole conditions created by the increasing popula-
and the decreasing pH. In addition, more acidand higher levels of LAB in summer than springer cheese coincided with lower levels of Enter-e and coliforms in the respective cheeses. Bothroups also disappeared rapidly from cheeseay and they underwent a more acceleratedheese made in March than the cheese made in
ci dominate in the cheese until 15 days of stor-nd lactobacilli become predominant in the 30--old cheese. Leuconostoc spp. were present atitially, but occurred more frequently in cheese. Lactococci were identied, mainly, as citrate
-
24 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
Fig. 4. Counts ge of An
fermentingLb. plantaruamong Leutively. The respect of and esterasfrom heat-milk cheeseas starter c(Xanthopou
3.2. Galoty
Galotyrimade from cheese is mdiffering fro
The milkit is salted (this time thto a skin balled. The cNowadays, skin bags. moulds, the
Retail chture contenis found at l(pseudomococci) are pL. monocytoKakouri, 20yeasts and this cheese
tisana90 Cbial p
(lactoNSLABd, primphilicucing Nf the Nlevels ot und
signiays of (log10 cfu/g; x SD) of lactic acid bacteria during manufacture and stora
. Lc. lactis subsp lactis, Ln. paramesenteroides andm were the most frequently isolated speciesconostoc spp. and Lactobacillus spp., respec-isolates of lactococci differed considerably intheir acidifying, caseinolytic, aminopeptidasee/lipase activities (unpublished data). Cheesestreated milk lacked the typical avor of raw, despite the use of Lc. lactis subsp lactis strainsultures, isolated from the traditional cheeselos et al., 2000a,b).
ri, PDO
Arto 85microto 8.0The posemesoprodpart ohigh did ngrew14 d is a soft cheese with sourish and brakish taste,ovine milk at the end of the lactation season. Theade in all regions of the country by processesm region to region.
is boiled and put in a clay pot, and after 24 h4% NaCl) and left for another two days. Duringe milk is mixed every 3 h. It is then transferredg. This is repeated several times until the bag isheese ripens for three months (Zygouris, 1956).the cheese usually ripens in barrels instead ofIn this case, in order to avoid the growth of
cheese surface, is covered with melted fat.eeses have a low pH (3.744.0) and high mois-t (av. 76.4%). The predominat population of LABevels of 8.0 log10 cfu/g and undesirable bacterianads, enterobacteria and pathogenic staphylo-resent at low levels (
-
E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 25
107 and 106 cfu/g, respectively. Enterococci were abun-dant (60.8% of the isolates), lactobacilli were found quiteoften (23.5%) and weisselae and leuconostocs were consid-erably less frequently (9.8% and 5.9%, respectively) isolated(Fourkiotis by coliformtent (brine microbial g
3.4. Kopani
Kopanispery taste mislands of Adardized buproducers. a small quacurd is draiolder cheeshand and focheese is thor blue greeballs are thethat the mocheese. Theis kneaded 1956).
In retailin 86% of t78%, and cThe cheese72% of the 105 cfu/ghe cheeses, yeastsmoulds were >103 cfu/g inoliforms were absent in 60% of the samples.
pH (in 74% of the cheeses 60%) and salt (in 52% of the cheesesed greatly. Lb. plantarum and Lb. casei subsphe dominant lactobacilli and enterococci and P.
were also isolated. Among yeasts, Pichia mem-s dominated while moulds were characterizedm commune (Tzanetakis et al., 1987).
alo Chanion, PDO
ft spreadable cheese. The milk is left to soure cheese is drained, salt is added at 1% and theeady for consumption. The product is made atrete island.ss composition of this cheese was foundllows (Papageorgiou et al., 1998): moisture.63% 4.67; fat in dry matter 54.03% 7.73;tent 14.23% 1.72; salt content 1.02% 0.38;.003; and pH 4.36 0.25. Salmonella and L.nes were not detected, coagulase positiveci were present in 6.45% of the samples andnd sulte-reducing clostridia were isolated% and 40.32% of the samples, respectively. Thentained high populations of coliforms, ente-sts and moulds and psychrotrophic bacteria,
c acid bacteria constituted the predominantroora. It was also shown that high qualitylo Chanion cheese can be produced using a
mixture of ewes and goats milk and 4% ofstarter culture.
Kawith mixtuducethe eGreecareas
Pl(510(Anifat 3238 Cleft t824unifobaskein a betwand ia woothe shthe c23 dthe wtrans
Thand icontemattmatunant enterdepeand low iing rlactomicrobial gwith sition30% and eacid coccicontrthrouactiv
4.2. K
KrorganertiesripenPossis a traditional Greek cheese of pasta lata typesant avor, manufactured from ovine milk or
ovine and caprine milk. The cheese was intro-eece from the neighboring Balkan countries atthe 19th century, initially to Thessaly, Central
where its manufacture was spread to other country.ine or mixtures of it with caprine or bovineilk are used for the cheese manufactures, 1991; Zygouris, 1956). The milk is curdlede coagulum is cut and it is usually cooked atcurd is cut in large pieces, and the baski is
in a cheese cloth and ripen over a period of20 C. The ripened baski is cut into small thinces, which are put into a basket (kanelo). Thepped into water (at 70 C), where it is turnedn order to facilitate the penetration of watere slices. The cheese thus becomes soft, elasticormity is achieved by its thorough mixing withdle. Then, the cheese is cut in pieces, formed to
spheres and introduced into the moulds, whereemains for 24 days. The cheese is salted everyntil it accepts 1214 saltings. The ripening in
room (18 C) lasts for at least 70 days; it is then to cold stores (02 C).
of the traditional fresh cheese is over 5.0reased slightly throughout ripening. Moisturege between 42 and 46%, fat content in dry
round 45% and the brine concentration of theese is around 3.0%. LAB constitutes the predomi-ora throughout cheese ripening (107 cfu/g) and
are present at quite high levels (104106 cfu/g),on the milk quality. Staphylococci, coliformsare found at levels from negligible to quitefresh cheese and decrease their numbers dur-g. In the fresh cheese (baski) pediococci andconstitute the 25% of the predominant LAB
and enterococci are the most abundant micro-50%). The proportion of pediococci is increasedogress of ripening and at 120 days the compo-
LAB microora is: pediococci 50%, enterococcictobacilli 20%. Selective isolates of pediococciocci were found to produce small quantity ofis makes them appropriate as adjuncts. Pedio-
more proteolytic than enterococci and mayo the acceleration of cheese ripening and avorir strong aminopeptidase and other proteolyticnpublished data).
yri (Possias)
ri or Possias has slightly sour taste andc properties very much affected by the prop-ine sediment, where the cheese is put afternifantakis, 1991). The wine sediment is called
which the cheese was named. The cheese
-
26 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
is made from ovine or caprine milk or mixtures of both,mainly in Kos island, Dodekanisos.
The milk from the afternoon milking is heated untilboiling and it is then transferred into swallow clay pots.The next morning, the milk fat risen on the milk surfaceis removed and the boiled milk is thoroughly mixed withfresh milk from the morning milking. The milk is coag-ulated with rennet (usually traditional), the curd is cutand then is transferred into baskets (tyrovolia) to drain for24 h. After that, the baskets are removed the surface ofthe cheese is salted and then is put in vessels. The cheese istransferred in cool places to ripen for 2030 days and afterthat it is removed from the vessels and put on shelves todry. The dry cheese is again put in the vessels and wine sed-iment is added to cover the cheese. In this environment thecheese ripens for one more week. At the end, the cheesesget a reddish colour of the sediment (Anifantakis, 1991).
The NSLAB constitute the predominant microora ofthe cheese and are found at similar levels on the cheesesurface and interior (Kalavrouzioti et al., 2004). The salt-tolerant ancheese surfpossibly duand/or the face (4.38 a2.49%, respeof the lactiwas compowith E. faeccheese intelactococci, bE. faecium addition, thcasei were isolates of were foundof the cheerespectively
5. Hard ch
5.1. Gravier
Gravieracheeses. It
its ne taste. Sheeps milk is mainly used for cheesemanufacture, but cows milk or mixtures of it withsheeps and goats milk are also used, the latter notexceeding 20%. The mature cheese has a smear rindand, usually, exhibits small or larger irregular openingsand could be characterized as slits of holes rather thaneyes.
The Greek Graviera was initially manufactured in thesecond decade of the 20th century. Nowadays, many typesof Graviera are traded and are distinguished from the nameof the region where they are produced (Graviera of Crete,Naxos, Mytilini, Tinos, Lamia, Kerkyra, Larissa, Dodoni andothers).
The milk is renneted at 3336 C with a quantity ofrennet able to curdle the milk within 2530 min. The useof starter is optional (Zygouris, 1956). The curd is cutin small pieces, the size of corn and cooked at 4850 Cunder continuous stirring. The curd is cut in pieces,each one enough to ll one mould. The cheese in themoulds is pressed for 24 h. After pressing, the cheese is
n she
rface sn thehs.owadaophilicse mane trad
contalationdant m
numb decree NaCeen 2re che% of
ced (Ta the vaent N
cheespecies urd als startnosto
Table 5Values (x SD sis (o-PAof Graviera Kr
fter salt
pH 5.38 Moisture% 1.92 NaCl-in-moi 3.05 % degradatios-CN -CN
Proteolysis (Lipolysis (AD
a Mean of thb In ppm of d the coliform microoras were less on theace than the interior by 1 log10 cfu/g. This wase to the effect of possia and not of the pHNaCl, which were similar for the cheese sur-nd 2.38%, respectively) and interior (4.38 andctively). Possia also affected the composition
c microora. Thus, the surface LAB microorased of lactobacilli and enterococci (50% each),ium predominating. The LAB microora of therior consisted of lactobacilli, enterococci andut lactobacilli predominated. Lb. plantarum and
were the most frequently isolated species. Ine species Lb. rhamnosus, Lb. casei and Lb. para-not isolated from the cheese surface, while noLb. buchneri, E. gallinarum and E. pseudoavium
in the cheese interior. The mean moisturese surface and interior was 54.4 and 55.6%,.
eeses
a, PDO
is the nest cheese among the Greek hard-is distinguished for its pleasant aroma and
put ois suing omont
N(meschee
Thmilk)popuabuntheirually
Thbetwmatu14.39redu
Atdifferof thecus sthe cact aLeuco
)a, of pH, moisture%, NaCl-in-moisture%, degradation of caseins, proteolyitis cheese made from raw ewes milk.
Days at 17 C
Before cooking After cooking Before salting (1 day) A
6.27 0.23 6.48 0.15 5.3 0.049 67.1 4.15 53.69 9.48 45.51 3.01 4
sture% n of caseins
0 0
o-PA) 7069.72 2774.35 V) 0.227 0.03 ree cheesemaking trials.l-glycine equivalents.lves in the salting rooms (1416 C) where italted by 3540 or even 60 saltings depend-
cheese height. The cheese ripens for 34
ys, the cheese milk is pasteurized and starter + thermophilic or fresh yoghurt) is used forufacture in modern dairies (Anifantakis, 1991).itional Graviera Kritis cheese (from raw ewesins Enterobacteriaceae and coliforms at high
s (Fig. 5). In addition, NSLAB constitute the mosticrobial group during ripening. They increase
ers in the cheese before salting and then grad-ase till the end of storage (6 months).l-in-moisture of the cheese was found to range.42 and 3.05% and moisture content of theese was 31.52%. At this time the 65.36% ands- and -CN, respectively, of the cheese wasble 5) by the native ora, mainly the lactic.rious steps of cheese manufacture and ripening,SLAB species were found (Table 6). In the curde lactococci predominated. The main Lactococ-was Lc. lactis subsp. lactis. Its high frequency inlows us to speculate that Lc. lactis populationer to drive the initial acidication of the curd.cs in the curd constituted a signicant part of
)b and lipolysis (acid degree value ADV) during ripening
ing (4 days) 45 days 90 days
0.075 5.38 0.2 5.07 0.0794.14 39.44 1.1 33.6 1.960.82 2.53 0.28 2.417 0.14
33.12 12.91 65.36 2.8812.15 3.94 14.39 2.77
8013.23 986.81 9016.8 1902.030.3315 0.212 0.5759 0.045
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E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 27
0
1
2
3
4
5
6
7
8
9
Before cooking Aft er cook ing 1 4 Time of ripenin g (days )
log 1
0 cfu
/gColiformsEnterobacteriacea e
Fig. 5. Populations (log10 cfu/g; x SD) of Enterocacteriaceae and coliforms throughout
lactic microora. Leuconostoc mesenteroides subsp. dextran-icum and Ln. pseudomesenteroides found in the curd, may begreatly responsible for the formation of small holes in thecheese body. Lb. paracasei subsp. paracasei, Lb. plantarum
Table 6Changes in numbers and kinds of NSLAB isolates during ripening ofGraviera Kritis cheese made from raw ewes milk (number from threecheesemaking trials).
Ripening time Species No. ofisolates
%
Before cooking Lc. lactis ssp. lactis 4Lc. lactis ssp. cremoris 5Lc. rafnolactis 2 47.83Lb. paracasei ssp. paracasei 1Lb. brevis 1 8.7Leuc. mesenteroides ssp. 3
45 days
90 days
and heterolow proporformation btively.
The prowith ripenicheese at 4erofermentripening. Thseems possnated over
Enterocopredominaning. Leuconcheese micrcarnobacter
e (unpaviera) and g ripedextranicumLeuc. pseudomesenteroides 6 39.13E. faecium 1 4.34Lb. paracasei ssp. paracasei 8Lb. paracasei ssp. tolerans 2
cheesGr
(>5.3durinLb. plantarum 7 89.29Lb. paraplantarum 1Lb. curvatus 1Lb. brevis 5Lb. buchneri 1C. piscicola 1 3.57Leuc. pseudomesenteroides 1 3.57E. durans 1 3.57Lb. paracasei ssp. paracasei 24Lb. plantarum 6Lb. paraplantarum 3 84.09Lb. biffermentans 1Lb. coryneformis 1 4.55Lb. brevis 1 2.27Lb. parabuchneri 1Leuc. mesenteroides ssp.mesenteroides
1 9.09
Leuc. mesenteroides ssp.dextranicum
1
W. confusa 1E. faecalis 3E. malodoratus 1
until the chactivity of thganisms. Pr(L. bulgaricution of diacwas relatedable microbwere increaquantities othe age of amounts fo
Additionmesophilic bacterium frgross comppH and thestarters weiforms were45 90
manufacture and ripening of Graviera Kritis cheese.
fermentative lactobacilli, found in the curd intions may play a minor role to the open bodyy gas production from citrate or lactose, respec-
portion of lactococci isolation was decreasedng and these organisms were not present in the5 days. On the contrary, the facultatively het-ative lactobacilli increased their frequency witherefore, their contribution to cheese ripeningible. Lb. paracasei subsp. paracasei predomi-the NSLAB in the mature cheese.cci participated at a quite low percentage in thet lactic microora throughout cheese ripen-ostocs constituted a low proportion of theoora throughout ripening and weissellae andia emerged at some ripening stages of theublished data).
cheese made from cows milk had a high pHbrine concentration ranged from 0.43 to 4.10ning for 180 days. The WS-N increased rapidly
eese was taken out of the warm room, due to thee increasing population of proteolytic microor-oline formation was mainly due to the starters, propionibacteria) activity and peak forma-etyl at the end of ripening in the warm room
to the rapid increase of almost all the desir-ial groups. The amounts of C2 to C6 fatty-acidssed throughout ripening for 180 days. Highestf fatty acids from C10 to C16 were recorded at3 months with palmitic acid being in highestllowed by myristic acid (Zerridis et al., 1984).
of combination of thermophilic and/orand thermophilic starters along with Propioni-eundereichii subsp. shermanii did not affect theosition of the ripening cheeses. However, the
enterococci counts in the cheeses made withre signicantly lower than the control and col-
not detected (Kandarakis et al., 1998).
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28 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
5.2. Kefalograviera, PDO
Kefalograviera is a hard cheese, with a rm body andmany holes or eyes, mild to medium piquant avor andcolour from60/40 cow/also be incoKefalograviGreece. As togy of manGraviera ch
When fuwith differstarter did nbut the lattethe low-fatfat contentuble nitrogadjunct culnitrogenou2003). The levels of acacetoin at 1had signicone than thtype of start(Kondyli et
5.3. Kefalot
Kefalotyby high harfrom sheepmay also be
The cheGreece cenof Greece wand it is tris manufactetc.).
The milkit is usuallycheese milsmall particunder contithe size of othey are thripening roB) for 12 where it is washed witto continue
The pH obe >5.0, thewas 38% microora pwas compo(Lc. lactis), lactobacilli mentative)Streptococc
Table 7Log microbial counts, pH, moisture % and NaCl-in-moisture % (x SD) ofOrinotyri cheese from raw sheeps milk.
a
robacterformsts
sture % l-in-moi
eans of t detect
f ripenmina
etaki, 1 Orin
nameing wterobald chhree mlog10 cse (Tabse wasnits ins in thted inher free leveing. Encocci, tly foucomple 8). Fi
chees pento
3-mo and tcalis, Eiformie freshLc. garh wereat em-mont. torquii).
Ladotyri, PDO
dotyri is a hard cheese, produced mainly in the islandytilini. It is considered as a type of Kefalotyri of goodty. The cheese is made from ewes or mixtures of ewesgoats milk raw at home or pasteurized in dairies.tyri is made by different methods but the overallology is very similar to that used for Kefalotyri man-ure. Ladotyri is preserved in olive oil, as indicated byme (ladi = olive oil; tyri = cheese). nearly white to slight yellow. A mixture ofsheep milk is usually used, but goats milk mayrporated in a proportion not higher than 20%.era is manufactured, mainly, in mountains ofhe name of the cheese is indicating its technol-ufacture has elements of both, Kefalotyri andeese (Anifantakis, 1991).ll-fat and low-fat cheeses were manufactured
ent starters, it was observed that the type ofot affect the composition of the low-fat cheeser received signicantly higher avor scores that
control (Katsiari et al., 2002). In addition, the affected the pattern of proteolysis, water sol-en was signicantly affected by the addition oftures and the production of low molecular masss compounds was enhanced (Michaelidou et al.,experimental cheese had signicantly higheretone at 90 days and acetic acid, diacetyl and80 days than the control low-fat cheese, whichantly higher levels of butan-2-ol and butan-2-e former cheeses at both sampling ages. Theer also affected the total levels of free fatty acids
al., 2003).
yri
ri (Kefali + tyri = head + cheese) is characterizeddness, salty taste and strong avor. It is mades or goats milk or mixtures of them. Cows milk
used.ese technology was introduced from Italy toturies ago. It is manufactured in various partsith technologies that differ from area to area
aded under the name of the region where itured (Crete, Naxos, Thessaly, Kefalonia, Epirus,
is partially skimmed (5.86.0% fat content) and pasteurized. Starter cultures are added in thek. After coagulation, the coagulum is cut intoles. The temperature is then raised to 4345 Cnuous stirring. The coagulum is cut into pieces,ne mould. The pieces are put into moulds anden put into press. The cheese is placed in theom (1416 C) for 24 h and then in brine (1820days. It is transferred again in the ripening roomgiven 2025 dry saltings. The cheese is nallyh brine and transferred into a cold store (24 C)
ripening.f the cheeses sold in retail shops was found to
percent moisture in the majority of the cheesesand the brine concentration 6.8%. The lacticresent in the fresh cheese from raw cows milksed of streptococci (S. thermophilus), lactococcienterococci (E. faecalis, faecium and durans),(obligately as well as facultatively heterofer-
and leuconostocs (predominantly Ln. lactis).i and lactococci were not isolated after the 15th
EnteColiYeaspH MoiNaC
a Mb No
day opredoTzan
Inlocalfollow
Enday-othe t7.76 cheecheepH uleveldetecin eit
ThripenLactoquenwas (Tablfreshby P.In thefoundE. faecorynin thatus, whicers ththe 3subsphioch
5.4.
Laof Mqualiand Ladotechnufactits naFresh cheese Three-months-oldcheeseb
iaceae 7.94 0.19 4.93 0.917.41 0.29 4.65 1.081.99 0.77 ND26.31 0.19 5.79 0.28
49.16 5.82 36.4 3.05sture% 4.13 1.83 6.64 1.85ve cheesemaking trials.ed.
ing; leuconostocs were also present among thent LAB of the 2-month-old cheese (Litopoulou-990).
otyri (Orinos + tyri = mountainous + cheese), a for Kefalotyri, made from raw ewes milk, theere observed (Prodromou et al., 2001).cteriaceae and coliform counts in the ten-eese, were high; their average counts inonth-old-cheese were smaller by 3.02 and
fu/g, respectively, than counts in the freshle 7). At the same time, the pH value of the fresh
high (>6.31 pH units) and it was reduced by 0.52 the mature cheese. Yeasts were counted at lowe fresh cheese (200 cfu/g) and they were not
the mature product. S. aureus was not detectedsh or mature cheese.ls of NSLAB were decreased signicantly withterococci predominated in the fresh Oritotyri.lactobacilli and pediococci were also less fre-nd in the fresh cheese. The NSLAB microoraex in both, 10-day and 3-month-old cheesefteen different species were characterized in thee, with E. faecalis being predominant, followedsaceus, Lc. lactis subsp. lactis and Lb. curvatus.nth-old cheese fourteen different species werehe predominant at decreased frequencies were:. faecium, Lb. paracasei subsp. paracasei and Lb.s subsp. torquens. There were also species found
cheese (E. faecalis var. liquefaciens, E. malodor-vieae, Lb. paracasei subsp. tolerans, Lb. curvatus)
not detected in the mature cheese and oth-erged as ripening progressed and were found inh-old product (Lc. rafnolactis, Lb. coryneformisens, Lb. plantarum, Lb. paraplantarum, Lb. homo-
-
E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 29
In cheese made from raw ewes milk microbial countswere measured at high levels (mean counts of coliforms1.8 105 cfu/g; yeasts 5.8 104 cfu/g; halotolerant bac-teria 1.5 105 cfu/g; psychrotrophs 1.3 106 cfu/g) andlactic acid bacteria constituted the predominant microora(mean counts 3.2 108 cfu/g and 4.1 108 cfu/g for cocciGram positive and catalase negative and lactobacilli,respectively). Enterococci predominated (58%) over lacto-bacilli (36.2%), lactococci, S. thermophilus and leuconostocs,the cheese pH was over 5 and its moisture and NaCl%content were 31% and 3%, respectively (Fista et al.,1996).
5.5. Manoura
Manoura is a hard traditional farmhouse Greek cheesevariety manufactured by producers of Sifnos island in theAegean sea (Cyclades complex) from raw ovine or a mixture(50:50) of raw ovine and caprine milk from local herds.
The cheese is made as follows: the milk is not heat-treated and it is not inoculated with a starter culture.Animal rennet deriving from the stomach of a young ani-mal is used to curdle the milk. The coagulum is cut intopieces, the size of a nut, and after a thorough stirring, it isleft to settle in the bottom of the vat. The curd is transferredto clean baskets (tyrovolia) to drain. The cheeses are saltedfor 23 days with coarse salt, on a different side each day,after removal from the basket and inversion. After that, thecheeses are removed from the baskets and put on beds ofstraw to dry for 34 months. The cheeses are then put intobarrels with red wine to soften for about a week (510days). They are then put into barrels and covered with wine
Table 8Species of lactOrinotyri.
Lactococcus Lc. garvieae Lc. rafnolacEnterococcusE. faecalis vaE. faecium E. hirae E. saccharolyE. malodoratEnterococcusWeisella parPediococcus LactobacillusLb. paracaseiLb. curvatus Lb. coryneforLb. coryniforLb. plantarumLb. paraplanLb. homohiocTOTAL
a Isolates frorococci. Isolatenterococci 11rococci 10 straW. paramesent
sediment for 1 day. The cheeses are afterwards moved toempty barrels, where they are kept until sold (Gerasi et al.,2003).
Changes in the microora of Manura from raw ovinemilk were sgeneral, thethe cheese ment had ancounts decrin the intewere reducmonths) anLactic acid bial groupscremoris, P.sei, frequenas starters bacteria prsella paramecontribute activities.
5.6. Xinotyr
Xinotyriraw goats Naxos (Cycdled in abouamount of starter. Thefor 23 h anis kneaded transferred
and tved an45 d
the thangeseningr popuophileningB, sign
therms werectively4 cfu/g
elich
elichloheesetion see miln 1 h. Td into ing, thimes. Tal timen con to dic acid bacteriaa isolated from fresh and three-month-old
% of the isolates
Fresh Three-month-old
lactis ssp. lactis 16.5 25.01.6
tis 1.5 faecalis 24.7 27.9r. liquefaciens 1.6
6.6 10.31.6 1.5
ticus 1.6 1.5us 1.6
spp. 1.6 amesenteroides 1.6 4.4pentosaceus 23.0 2.9
paracasei ssp. paracasei 1.6 8.8 ssp. tolerans 1.6
11.6 mis ssp. coryniformis 3.2 2.9mis ssp. torquens 8.8
1.5tarum 1.5hii 1.5
100 100
m RA, M17 agar and CAA. All isolates from CAA were ente-es from M17 agar were; fresh cheese: lactococci 11 and
strains; three-month old cheese: lactococci 18 and ente-ins. Isolates from RA were lactobacilli, P. pentosaceus anderoides.
days remofor 30
Incal cin riphighethermof ripNSLAto theyeastrespeas 10
5.7. M
MThe clacta
Thwithiferredrain34 tseverwoodthe sutudied during ripening (Gerasi et al., 2003). In various microbial groups developed better onsurface than in the interior. Red wine treat-
inhibitory effect on their growth and microbialeased more rapidly on the cheese surface thanrior (Fig. 6). Enterobacteriaceae and coliformsed sharply during ripening on a straw bed (3d they were not detected in mature cheese.bacteria predominated over the other micro-
throughout ripening. Ln. mesenteroides subsp. pentosaceus and Lb. paracasei subsp. paraca-tly found in maturing cheese, could be usedto make this cheese. Moreover, the lactic acidedominating in mature cheese, such as Weis-senteroides, Lb. bifermentans and Lb. brevis, mayto cheese ripening though their biochemical
i
is a farm cheese variety, manufactured frommilk from indigenous breeds in the island oflades complex). The milk is renneted and cur-t 24 h at room temperature. Sometimes, a smallcheese whey from the previous day is used as
curd is transferred in a cheese cloth to draind then dry salt at 1.5% is added and the curdfor the uniform dispersion of salt. The curd is
into plastic truncated conical moulds for 34urned over daily. After that, the moulds ared the cheese is left to ripen on wooden shelvesays (Bontinis et al., 2008).raditional cheese the following microbiologi-
were observed (Bontinis et al., 2008). Early (6-day), the cheeses contained signicantlylations of mesophilic LAB and lactococci than
ic LAB and streptococci. During the later stages, there were major declines in populations oficantly higher for the mesophilic as comparedophilic part of the NSLAB ora. Enterococci and
counted at quite high levels (105 and 107 cfu/g,), early in ripening and declined to levels as low
for enterococci, and 103 cfu/g for yeasts.
loro (melipasto)
ro is the traditional cheese of Limnos island. is made from raw ewes milk at the end of theason.k, warm as it is after milking, is coagulatedhe curd is cut into very small pieces and trans-
baskets for draining on wooden shelves. Duringe cheese in the baskets is reversed every 1 h foren hours after curdling the cheese is dry saltedes on each side. Then, the baskets are put in anstruction like a cage, hung from a tree, underry and the baskets are inversed every day. After
-
30 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
Table 9Mean log microbial counts (log10 cfu/g; x SD) during storage of Manouri cheese in spring and summer (mean of three cheesemaking trials). Means in thesame row with different superscripts were signicantly different (P < 0.05).
Days at 4 C
Microbial group Sample from Season 0 (cheese after draining) 5 10 20
Total aerobic count Surface Spring 4.25a 0.24 6.01b 0.44 6.69bc 0.96 7.32c 0.64Interior Spring 4.73a 0.75 5.10ab 1.25 5.82abc 0.68 6.87c 0.84Surface Summer 4.83a 0.03 6.45b 0.99 7.48bc 0.65 8.34c 0.66Interior Summer 4.91 0.42 6.25 1.22 6.31 0.70 6.33 0.75
Enterobacteriaceae Surface Spring 2.67a 1.12 3.04a 0.04 5.65b 1.25 7.26b 1.59Interior Spring 0.00a 0.00 2.93b 0.14 5.14c 1.13 5.37c 1.18Surface Summer 3.16a 1.25 5.69b 0.95 7.14bc 0.82 8.32c 0.56Interior Summer 1.59a 0.51 2.94a 1.94 5.32b 1.09 5.89b 0.77
Coliforms Surface Spring 2.28a 0.64 3.45ab 0.74 4.39bc 0.99 5.34c 1.15Interior Spring 0.98a 0.68 3.29b 0.71 2.41ab 0.39 1.78ab 1.23Surface Summer 3.06a 0.82 5.12b 0.79 6.59c 0.45 8.32c 0.56Interior Summer 1.17a 0.81 2.47a 0.99 5.65b 0.53 5.75b 0.97
Total LAB Surface Spring 5.07a 0.53 5.52ab 0.47 5.76ab 0.54 6.54b 1.04Interior Spring 4.98 0.55 5.10 0.48 5.05 0.18 5.17 0.46Surface Summer 4.13a 0.65 5.01ab 0.87 5.71bc 0.72 7.05c 0.65Interior Summer 5.17 0.39 4.76 0.71 5.23 0.23 5.16 0.32
Lactococci Surface Spring 3.97a 0.76 5.52ab 0.47 5.78bc 0.56 7.02c 1.18Interior Spring 3.08a 1.60 4.55ab 0.48 5.09ab 0.11 5.36b 0.41Surface Summer 4.16a 0.40 4.73b 0.42 6.11b 0.67 8.21c 0.38Interior Summer 4.45a 1.00 4.95a 0.26 5.10a 0.25 6.40b 0.42
Enterococci Surface Spring 2.90a 1.39 4.97ab 1.09 5.15ab 1.76 6.42b 1.45Interior Spring 2.25a 0.67 2.65a 0.97 3.30ab 1.21 4.66b 1.05Surface Summer 2.50a 0.10 4.26b 0.23 5.30c 0.46 7.17d 0.35Interior Summer 2.86ab 1.18 2.54a 0.99 4.26bc 0.35 4.75c 0.69
Yeasts Surface Spring 4.90 1.06 5.30 0.23 4.93 0.25 6.46 0.81Interior Spring 4.65 1.01 4.85 0.57 4.67 0.58 5.40 0.37Surface Summer 3.53a 0.22 5.41b 0.52 5.65b 0.97 6.81b 1.09Interior Summer 2.41a 1.14 4.65b 0.50 5.22b 0.10 5.34b 0.32
Staphylococci Surface Spring 4.06a 0.98 6.21b 0.96 5.92b 0.12 6.68b 0.76Interior Spring 3.92 1.42 3.89 0.52 4.15 0.69 5.12 0.16Surface Summer 3.64a 0.52 5.49b 0.29 5.63b 0.45 6.64c 0.12Interior Summer 3.37 1.03 3.43 0.83 4.12 0.85 4.39 0.47
Fig. 6. Counts (log10 cfu/g; x SD) of different microbial groups at three main points of ripening of Manura cheese.
-
E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32 31
45 days the baskets are removed and the cheese wheelsare left in the cage for further drying. When they are hardenough, the cheeses are washed with sea water and driedwith a cloth.
The meaits moisturecentration various micand 7.11 (lasisted, mainLb. paracase
6. Whey ch
6.1. Manou
Manourwith myzitdelicate tex1991; ZygoWestern Mbeen madetion of Batand ovine the whey oalthough acheesemakmanufacturmaking myis lower tha
Selectedstorage for (Lioliou et aincreased (Plevels in chspring. Moron the cheesummer (Tacontent (2.5the growthobacteriaceaBaird-Parkewas a greathansenii anisolates of Eextent- yeaNH2) conteduring storalso seemsextent (acidity of the stisolates of Eand phosph
6.2. Myzith
Myzithrheating the1956). The wa temperatter quality
(in a proportion of 35%) when it obtains a temperatureof 6570 C. Salt (11.5%) may also be added to the wheywhile it is heated.
inomy
is typg thes and
d. For tfactur
t is thehe chehich aen for
inotyr
notyris madetter ws prodeatede clothhen puree mos in thi40 an8 (unp
ict of
one of ) has
peopnce oistics o
ences
oyiann, 2005. Pm infantakis, E.iry Com, T., Vbinson, bacilli an, 6877is, T.G.,ysicochg and stoilk. Int. J., TzaneLadotym on Laldhovencieties, n Leeuwotis, A., icroorangress o, E., Litody of Mand Sifnn pH of ve cheese samples was 4.49 0.2, content 31.4% 1.5 and the mean brine con-
was measured at 5.8% 2.1. The log counts ofrobial groups ranged between 1.63 (coliforms)ctococci). The NSLAB of the mature cheese con-ly, of Lb. paraplantarum (56% of the isolates) andi subsp. paracasei (34%) (unpublished data).
eeses
ri, PDO
i is a whey cheese that has many similaritieshra, but it is creamier and less salty, with ature and superior sensory quality (Anifantakis,uris, 1956). Manouri is a traditional product ofacedonia, in northern Greece, and it has long
from the whey obtained during the produc-zos, a semihard cheese derived from caprinemilk. Today, Manouri is also produced fromf cheeses made solely from ovine milk, and
whey cheese, it is the main product of thising process. The overall technology of Manourie has much in common with the process forzithra, but the nal moisture content (5060%)n that of the latter cheese.
microbial groups were counted throughout20 days at 4 C in batches of Manouri cheesel., 2001). The counts of all the microbial groups
< 0.05) throughout storage and reached highereeses made in summer than in those made ineover, the micro-organisms developed betterse surfaces than in the interiors, especially in theble 9). The pH (6.787.33) and salt-in-moisture33.72) of the cheeses did not seem to affect
of bacteria and yeasts. The isolates of Enter-e were mainly Hafnia, while the isolates fromr medium were mainly staphylococci. There
diversity of yeasts species, but Debaryomycesd Pichia membranefasciens predominated. Thenterobacteriaceae, staphylococci and-to a lessersts were proteolytic, but the free amino acid (N-nt of the cheese did not increase signicantlyage (136.4225.2 mg/kg glycine equivalent). It
that milk fat was not degraded to any great degree value 0.090.19) by the lipolytic activ-rains. The main enzymes detected in selectednterobacteriaceae were leucine aminopeptidaseohydrolase.
ra
a is a traditional Greek whey cheese made by whey at a temperature of 8892 C (Zygouris,hey heated under continuous stirring to obtain
ure of 8892 C within 4045 min. For a bet-of Myzithra, whole milk is added to the whey
6.3. X
Thdurinsheepislanmanuand iand trels wto rip
6.4. X
Xithat iof buFor itit is hcheesand tor thlevel% is is 14.
Con
Ntakisotherinueacter
Refer
AmbadN.fro
AnifanDa
BintsisRoto95
BontinPhinm
Fista, Gof siuVeSova
FourkiMCo
Gerasistuislzithra, PDO
e of Myzithra is made from the whey obtained production of Kefalotyri and Graviera from
goats milk. It is a traditional product of Cretehe production of Xinomyzithra, Myzithra is rsted, which is cooked and drained more severelyn pressed for one week. Salt is added after thatese is mixed. Then, the cheese is placed in bar-re transferred in rooms of 510 C temperature
a period of two months.
i (Klotsotyri, giza, artymi or prentza)
(xyno = sour + tyri = cheese) is the cheese type from the whey obtained during the productionith a pleasant sourish and salty pleasant taste.uction, the whey is put into a copper vat, where
until curdling. The cheese is left to drain in a for several hours. The drained cheese is saltedt in either barrels or skin bags or tins for twonths to ripen. The NSLAB are detected at lows cheese. The cheese pH is
-
32 E. Litopoulou-Tzanetaki, N. Tzanetakis / Small Ruminant Research 101 (2011) 17 32
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Microbiological characteristics of Greek traditional cheeses1 Introduction2 Cheeses in brine (Pickled cheeses)2.1 Batzos cheese, PDO2.2 Feta cheese, PDO2.3 Touloumissio cheese (Touloumotyri)2.4 Kalathaki of Limnos, PDO2.5 Sfela (Feta of fire), PDO2.6 Teleme
3 Soft cheeses3.1 Anevato, PDO3.2 Galotyri, PDO3.3 Katiki, PDO3.4 Kopanisti, PDO3.5 Pichtogalo Chanion, PDO
4 Semi-hard cheeses4.1 Kasseri, PDO4.2 Krassotyri (Possias)
5 Hard cheeses5.1 Graviera, PDO5.2 Kefalograviera, PDO5.3 Kefalotyri5.4 Ladotyri, PDO5.5 Manoura5.6 Xinotyri5.7 Melichloro (melipasto)
6 Whey cheeses6.1 Manouri, PDO6.2 Myzithra6.3 Xinomyzithra, PDO6.4 Xinotyri (Klotsotyri, giza, artymi or prentza)
Conflict of interest statementReferences
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