NICOLAE Ion Member of the Academy of Romanian Scientists...

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BULLETIN OF SCIENTIFIC INFORMATION - NR.22 / 2011

Rector’s Allocution

We have the special pleasure to let you know that the Review of our University,„Bulletin of Scientific Information”, having ten years of consecutive issue, it achieved the recognitionof the National Council for Scientific Research in Higher Education (NURC), beingcomprised in the category „National Reviews – C Category”.

So, the Bioterra University review „Bulletin of scientific information” works as a real platformfor the information and exhibition of the most recent and valuable research in the agricultual field andconnected sciences (food industry, agro-tourism, ecology, agricultural economics etc.).

This way I express my gratitude the contributors to our review, authoritative academic and univeritarynames of whose studies are found in the selection done by the scientific board of the review, co-workers withwhom we have strong relations of partnership and mutual support in the development and course of someconjointed research projects.

I wish to the review many and consistent issues.

Prof. Floarea Nicolae, Ph.D.Rector of the Bioterra University Bucharest

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Sumary:

» Study concerning the cultivation and conServation optionS of

leontopodium alpinum SpecieSARISTOTEL Cristina, ELISEI Angelica

Faculty of Food Control and Expertise, 4th grade

Bioterra University of Bucharest05

» malolactic fermentation mlf: again in SpecialiStS’ attention with new

technical-Scientific approacheS! aSpectS of the white wine making (1)CROITORU Constantin


» malolactic fermentation mlf: again in SpecialiStS’attention with new

technical-Scientific approacheS ! aSpectS of the red wine making (2)CROITORU Constantin


» Study of Some tomato varietieS originating in england in the

pedoclimatic condition of South-eaStern romania, reSpectively Braila

countyGALAn C`t`lin, BOLOLOI Mihaela, EREMIA Florentina


» proSpectS for development of rural touriSm in romaniaAndROnE Roxana, POPARLAn Alina Maria, MIhAILA daniela Fanuta, MARICA Mariana daniela


» the concept of food Security in europeSTOICA Valeri2, ATUdOSIEI nicole Livia1, AndROnE Roxana1, EREMIA Florentina1

1Bioterra University of Bucharest2Academy of Agricultural Sciences and Forestry of Bucharest

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Introduction

» Leontopodium alpinum is a species of aspecial beauty and it represents the srarestspecies of the whole Romania’s mountain flora.Its area of origin is the calcareous plateaus andmeadows from the steep and sunny slopes.In Romania the species can be sporadicallymet in the Maramureului, Vrancei, Bucegi,F`g`ra], Retezat, Bihor, Cozia, and Ceahl`uMountains. The highest density is in Rodnei

Mountains where is was established theprotected area „ Natural Reservation of PiatraRea” having a surface of about 50 ha.Leontopodium alpinum forms a rosette ofleaves at the ground surface from the middleof which it raises a short stem of about 5-20cm, bearing a star-shaped inflorescencecovered with silver hairs which gives aspecial aspect. Depending on the annual thermal regime, itblooms in July-August (Picture No. 1 and 2).


Study concerning the cultivation and conServationoptionS of leontopodium alpinum SpecieS

Aristotel Cristina, Elisei Angelica Faculty of Food Control and Expertise, 4th grade

Bioterra University of Bucharest

Abstract: The research has the main goal to find some methods and techniques to cultivate andpreserve „in situ” and „ex situ” the species Leontopodium alpinum, perennial, herbaceousspecies, Genus Leontopodium Cass., Family Asteraceae, that has as development area the alpineplateaus of the Carpathian Mountains. In the last century the environment conditions degradation by pollution and excessive grazing,and also by harvesting the flower without control it leaded to the destruction of natural habitatsand dramatic restriction of this species habitat development.For this reason in Romania since 1933 Leontopodium alpinum has been declared a naturalmonument and it is also today protected at the national and international level with protectedspecies status. Due to this status the collection of specimens from the wild is strictly prohibited,that makes more difficult the study of the species in the laboratory conditions in order to elaboratesome culture technologies links.

Key Words: Leontopodium alpinum, protected alpine species, mountain flora.

Picture no. 01 - leontopodium alpinum in its native habitat

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Materials and Methods

There were used 53 plants of Leontopodiumalpinum grouped in 10 pots. The plants wereobtained from seeds harvested from the nativeenvironment and cultivated in greenhouseconditions (Picture No. 02) by the onlyedelweiss grower from Romania, NicolaeTiron.

For the transplant into the naturalenvironment there were chosen morelocations from din „ Bucegi National Park”,located on the mountain about 1500 maltitude, in remote places, inaccessible and farfrom tourist routes (Picture No. 03).

Due to the steep slopes and rough terrain thebatches of plants were hand-carried (PictureNo. 04)..

Using a mini-shovel the soil has been dug andloose to a depth of 10-20 cm for plant rootsto penetrate more easily. Extraction of theplants from pots has been performed carefullyin order to protect the week roots. In order toreduce the physiological stress due to thetransport, extraction from pot was performedat the planting place.

The specimens were planted at a depth of 8-12 cm in areas with predominantly calcareoussoil (Picture No. 05).


Picture no. 02 - production of seedlings in greenhouse conditions

Picture no. 03 - Selection of locations from Bucegi national park

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After planting the plots were wateredthoroughly with spring water to ensure goodcontact between the roots and soil.Eco-biological parameters of the batches ofplants were monitored from July 2 toSeptember 27, 2010, when the plants enteredin dormancy (Picture No. 06).

Results and discussions

By the proposed objectives the study was ableto effectively contribute to the conservationof rare plant species existing within protectedareas, by initiating procedures aimed atgrowing opportunities „in situ” and „ex situ”. The plants transferred in the native habitatwere monitor to quantify their viability rate,development rate, ability of flowering andpropagation, as well as to prevent the eventual

risk factors that could endanger the futureexperiments.

Conclusions

» the percentage of attachment

accommodation of the plants at thepedoclimatic conditions of the native habitatwas 93%, the plants going on with thebiologic circle and succeeding even to bloomfrom the very first year;» the result of the project van determine andencourage in the future the increasing of thecivic attitude of the people who love the naturein their desire to grow edelweiss; » also, this action can become a smallbusiness for the students who wish to getsome extra money without affect the flora andenvironment


Picture no. 04 - transport of plants at the elevation 1500

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.References

http://www.edelweissgrowers.com/

http://ro.wikipedia.org/wiki/Floarea-reginei

http://www.gradina-online.ro/Floarea_de_colt_A3668.html

http://atlasbotanic.florisicadouri.ro/

http://herbarium.biology.colostate.edu/onlinedatabases.htm

MOHAN Gheorghe, ARDELEAN Aurel -

Atlas botanic –Alfa All Publishing House,Bucharest, 2010

IONEL Adrian, MANOLIU Alexandru,ZANOVSCHI Valeriu - Cunoasterea siocrotirea plantelor rare - Ceres PublishingHouse, Bucharest, 1986

SIMIONESCU Ion - Flora Romaniei -Albatros Publishing House, Bucharest, 1974


Picture no. 05 - transplant in the natural environment of the leontopodium alpinum plants

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Picture no. 06 -plants state at the end of vegetation

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Introduction

Malolactic fermentation (MLF) in white winemaking remains a topical issue for bothpractitioners and researchers, in the sameextent as alcoholic fermentation (AF). Thenumber of producers choosing this process ofbiological reduction of the high acidity of thedry white wine using selected malolacticbacteria (SMB) and adequate nutrients is onthe rise. Very recent studies show that theinitiation stage of MLF (in the must alongwith the onset of AF, when the AF wasperformed in proportion of 50 - 60%, in thefinal stage of the AF or immediately after thefinalization of AF of the must) is crucial forboth the success and completion of theprocess and the insurance of the biologicalstability of the fermentation environment inwhich this process took place.The climate of the last period overturnedRomania’s territory, and the heavy rains haveproduced and will produce changes to the vinesin this active period of vegetation.

We may end up with grape harvest for whitewine varieties (I primarily think of Chardonnayand less of others) which excel through a toohigh acidity and a low concentration in sugars,which will require the application oftechnological solutions in order to reduce thevalue of this important analytical and sensoryparameter.I am taking into account the young plantationsof Chardonnay from Reca] and from Jidveithat produced wines gallooned with goldmedals at some of the latest prestigiousinternational competitions. In this context, anew approach to the issue of reducing theacidity in musts or in wines from whitevarieties through the biological pathway ofMLF seems appropriate.

Materials and methods

Knowing the evolution of the mainparameters (the SO2 concentration, thetemperature, the pH, ...) which act in theprocess of white wine production on a must


malolactic fermentation mlf: again in SpecialiStS’ attention with

new technical-Scientific approacheS!aSpectS of the white wine making (1)

croitoru constantin


Abstract: Malolactic fermentation (MLF) in white wine making is a new topic both for producersand for researchers, in the same extent as alcoholic fermentation (AF). The number of producerschoosing this process of biological reduction of the high acidity of dray white wines using selectedmalolactic acid bacteria (SMB) and adequate nutriments is on the rise. Very recent studies showthat the stage of MLF starting (by co-inocculation, by premature inoculation, by sequentialinoculation and staggered inoculation) is of top importance for the achievement and completionof this biological process and also for the biological stability ensurance of the resulted wine.

Key Words: white wine, malolactic fermentation, Sodinal Company.

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or a wine which requires a reduction of thetitratable acidity is very important in choosingthe commercial preparations of SMB andtheir compatible nutrient in order to guaranteeoptimal conditions to achieve MLF.» aspects of the So2 content. In white wineproduction, the action of sulphur dioxide usedwhen pressing the destemmed and crushedgrapes (derived from a healthy harvest ofChardonnay) is homogeneous because thetreated environment acts like a singlehomogeneous liquid phase; during setting ofthe resulted must after its separation from themarc, sulfur dioxide concentrations slowlyevolve according to its duration of action(table 1); it is noticed that after 3 days at 12°C, free SO2 concentration is 23 mg/l for aninitial dose of SO2 of 80 mg/l and only 10mg/l for the initial intake of 40 mg/l; reportedvalues show that the end of the setting of themust free SO2 concentration is approximately25 – 30 % of its original concentration whichwas administered at the pressing process.If it is envisaged that the fraction of molecularSO2 is the most active fraction of free SO2on the microorganisms, this fraction is evenmore important as the pH is lower and thetemperature and the alcoholic level are

higher; for example, for a pH of 3,1 and atemperature of 12 °C, a must which contains10 mg/l of free SO2 will have only 0,24 mg/lof molecular SO2 (according to the formulaestablished in the literature and tables ofvalues offered by professional institutions ofspecialist).Depending on the SMB strains and theircellular physiological state, it was found thatthe bacterial cells have a variable sensitivityto molecular SO2 concentration, with acritical limit allowed of 0,30 mg/l. In acomparative experiment on a must ofChardonnay kind, it was signaled a bacterialcell mortality after sowing with a strain ofSMB which varied depending on themolecular SO2 concentration of the sownmust; a drastic reduction of the bacterialcolony from the sown must was noticed inless than three days at a concentration of themust in molecular SO2 of 0,35 mg/l and inless than six days at a concentration of thesame must in molecular SO2 of only 15 mg/l.This simple experiment demonstrated that thesuccess of an early bacterial sowing with astrain of SMB depends largely on theconcentration in molecular SO2 in white winemaking (table 2).


analysed

parameters

Setting period

(hours)

So2 administered in the pressing

process

40 mg/l 80 mg/l

Free SO2 (mg/l) 244872

121010

302523

Free SO2 proportionfrom SO2 administered

(%)

244872

302525

383129

Table 1

the evolution of concentration in free So2 during setting of a chardonnay must (ph 3,1)

at a temperature of 12°c for two different levels of sulphitation

(Gerbaux V. and Briffox C., 2010)

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» aspects of the temperature evolution.

Relatively recent studies (Gerbaux V. et al.,2008) have shown that the implantation of astrain of SMB in a dry white wine with higheracidity occurs in optimal conditions at atemperature of 14 – 16 °C than for a slightlyhigher temperature of 18 – 20 °C. In case of earlybacterial sowing with a strain of SMB during AFof the must (whether or not derived fromChardonnay), the thermal impact on the bacterialimplantation heavily depends on the pH value ofthe fermented environment (table 3).

» aspects of the ph evolution. Recentresearch in an already presented doctoral thesis(H. Akin, 2008), led by Professor PierreStrehaino from the University of Toulouse, ona Chardonnay must with 205 g/l sugar, an initialpH of 3,1 and an assimilable nitrogen content of210 mg/l (of which 70 mg/l ammoniacalnitrogen and 140 mg/l amic nitrogen), whereAF was conducted at 18 °C in the presence ofselected strains of yeast, have shown that: in thefirst stage of 4 days, the pH decreased from3,10 to 3,03 and this decrease of the pH value


duration

(days)

molecular So2 concentration

< 0,05 mg/l 0,15 mg/l 0,35 mg/l

0 2,2 x 106 2,2 x 106 2,2 x 106

1 2,5 x 106 9,3 x 106 3,8 x 106

3 5,1 x 106 1,8 x 106 78

6 1,1 x 106 5 5

Table 2

the influence of molecular So2 concentration of chardonnay must (ph 3,1 and 15 °c)

on the implantation of a SmB population (viable cells/ml) after setting

(Gerbaux V. ]i Briffox C., 2010)

Table 3

the influence of the thermal regime on the SmB population after 6 days

of sowing and on the mlf implementation period

(sowing with a SmB population of 1 x 106 cells/ml at a must density of 1020 - 1030 g/l)

(Gerbaux V. ]i Briffox C., 2010)

must ph and the analyzed

parameters

temperature

16°c 18°c 20°c

chablis must 2009

(ph 3,11)

SMB (cells/ml) 9,5 x 104 6,0 x 104 1,7 x 104

FML period(days)

38 31 27

Bourgogne must 2009

(ph 3,26)

SMB (cells/ml) 6,0 x 106 8,6 x 106 1,0 x 106

FML period(days) 31 28 24

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is explained by the nitrogen assimilation whichis available during the growth phase of theselected yeast cell; from the fourth dayonwards, the pH value progressively increasedto reach the final value of 3,21 and this increaseof pH value is due to the proper fermentativeprocess wherethrough the gradual increase inalcohol concentration modifies the ionicbalance which has repercussions on the pH aswell; the increase of pH was more significantas the AF has advanced to the point of approx.80 %, after which these increases wereprogressively reduced until the completion ofthe fermentative process.The pH evolution during the AF of the musthas to be considered when choosing thebacterial sowing stage in case of severalvarieties of white musts (pre-eminently fromChardonnay) with high acidity. Choosing theSMB strain will have in view that the cellulargenetic heritage and the physiological state ofviable strain cells will exert a high enoughtolerance even towards the smallest variationsof pH (in case of approaching to the criticallimit of pH tolerance value for bacterialstrains of SMB cells, even a variation of only0,05 pH units can not be ignored !). Theinfluence of pH on molecular SO2

concentration (and of temperature as well)will also considered.


i. the SmB selection

Distribution companies can provide wineproducers compatible SMB and appropriatenutrients in order to ensure the achievementof MLF with the minimum requirementslisted above. Such SMB are presented in table4 and are also distributed by SODINALCompany.» the nutrient selection. Along with therecommended SMB from table 4, there isalso a complex nutrient which is compatiblewith these selected bacteria, calledFERMOPLUS MALOLACTIQUE. Thisnutrient is distributed by SODINALCompany and is corresponding to the aspectslisted below.

» the opportunity to use a nutrient at mlf

of wines. The nutrient should be used onlytowards amelioration, survival and growth ofbacterial cells in order to accelerate the kineticsof MLF in wine and not to develope these


Table 4

Strains from the Biolact range

important parameters

in the inoculation

phase

Biolact

acclimatée

Biolact

acclimatée

pB1025

Biolact

acclimatée

4r

Biolact

acclimatée

Bm

Biolact

cwr

Number of strains 3 1 4 2 1

Minimum pH 3,1 2,9 3,2 3,0 3,2

Temperature limit (°C) 18 15 18 12 -

Alcoholic degree (% vol.)

14,5 14 14 14 < 14

Min. temp. of inoculum 18 15 15 12 17

Total SO2 (mg/l) - - - - < 40

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unwanted bacterial species capable of causingvarious diseases. It is recommended the beused in all technology situations which requirethe finalization of MLF of wines. Typically, therecommended optimal dose is of 30 g/hl whichis administered in SMB leaven plus apreliminary dose of 5 g/hl administered in themust (wine) which is subject to biologicaldeacidification through MLF.» the characteristics of a mlf effective

nutrient. It has to be a nutrient with acomposition based on amino acids, vitamins,polysaccharides and mineral salts obtainedfrom the enzymatic and thermal degradationsof the autolysed yeast cell walls. It has toexercise the ability to rebalance the nutritionalenvironment exhausted by the yeasts after AF.It should be able to reduce the latency timewhich is necessary for the MLF onset. It hasto have a reasonable price in relation to itsquality.» the composition. Considering its complexcomposition, the product FERMOPLUSMALOLACTIQUE is a nutrient which is ableto:l rebalance the nutritional deficiencies ofthe wine intended for biologicaldeacidificationl to ensure the reduction of the latencytime at MLF onsetlmaintain a high bacterial cell viabilityuntil the completion of MLF

The composition of this product consists of amixture of 70 % cell walls, from autolysedyeasts, degraded through a thermo-enzymaticprocess (which provides all the nutritionalresources required for the performance ofmetabolic and fementative functions of SMB)and 30 % potassium caseinate.» packaging and coding. The product,FERMOPLUS MALOLACTIQUE, is sold inlarge bags with a net weight of 5 kg and hasthe code 001253.

ii. comparative study on the state of mlf

onset in white wine making

Literature (Croitoru C., 2009; Gerbaux V. ]iBriffox C., 2010) envisage 4 stages to MLFonset in white wine making, which will bediscussed below.» triggering through co-inoculation. Thisco-inoculation technique consists ofsimultaneously wine sowing with selectedyeast suspension and also with the SMBleaven. This technique confirmed its validityin the case of musts from very healthy cropswith a pH value close to 3,2 but denied theopportunity of its application in the case ofsome musts with pH values between 2,9 and3,15. Using this technique, the completion ofMLF may be extended up to 2-3 months. Otherspecialists opt for a „staggered co-innoculation” of the SMB leaven after 24 - 48hours from the administration of selected yeastsuspension (corresponding to the pH reductionperiod, as explained above). This technique is very dependent on both thespecificity for the metabolized substrate (malicacid) of selected SMB strain as well as thecompatibility of the nutrient used in relation tothis strain. In addition, it is compulsory that theadministered dose of SO2 does not exceed 30mg/l which, in the set must, would correspondto a content of free SO2 situated below thelimit of 10 mg/l which would correspond to aconcentration of molecular SO2 below 0,2mg/l which would affect in a lesser extent thebacterial cell viability.» triggering mlf through precocious

inoculation. This technique of precociousinoculation with SMB applies to the stage inwhich the duration course of the AF of themust was exceeded with 50 %, correspondingmore precisely with the stage in which thedensity of the must dropped around 1030 -1020 g/l. Even under these conditions, theSO2 dose administered in the destemming andcrushed grapes shall not exceed 40 mg/l when


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sowing with SMB leaven is required in a whitemust (suppose from Chardonnay kind) inwhich the pH is lower than the value of 3,2. This SO2 dose has to assure the protection ofthe must against oxidation, so that it mustoriginate from a healthy harvest. By applyingthis technique selected yeast cells will have thenecessary time through exercising theirmetabolism to produce those carbonylcompounds which are able to fully combine thefree SO2 content of the fermentativeenvironment; under these conditions, thesuccess of bacterial sowing is significantlyincreasing, especially because the stage whenSMB leaven is administered also correspondswith that maximum increase of pH previouslyanalysed. This technique is based on a slow butcontinuous metabolization of malic acid (andpossibly of citric acid) by the SMB leavenenriched with the compatible nutrient from themust which was subject to biologicaldeacidification through MLF but which wasalso previously „fortified” with the samecompatible nutrient for SMB (as shownabove). By exercising their metabolism, SMBwill not be able to convert to sugarmetabolization because the selected yeasts willcomplete this metabolization before the SMBwill complete the malic acid metabolizationand possibly citric acid metabolization.Comparative experiments conductedinternationally have shown that by applyingthis technique the sensory profile of theresulted wines changes in a favourablemanner and that there is no risk of unwantedbacterial deviations. However, this techniquehas the disadvantage that requires dailymonitoring of both of the fermentationprocesses, AF and MLF, in order to make surethat none of them will slow down or stop; thismonitoring includes both rigorous and dailycontrolling of the density which expresses themetabolic rate of sugars per unit time (e.g. in24 hours) and also acidity diminution (harderto follow, if we consider that the majority of

selected yeast strains also produce relativelysmall quantities of succinic acid and lacticacid, D and L forms of which will bediscussed in the next issue) and the evolutionof MLF (by significant daily control of theevolution of malic acid metabolization). » triggering mlf through sequential

inoculation. This technique of sequentialinoculation is applied in the final stage of themust AF (at a course stage of over 90 %). Thistechnique is recommended when the wine stillcontains 10 – 12 g/l fermentable sugars. Manyprofessionals opt for this technique because thefermentative environment becomesincreasingly rich in natural nutrients due toexorbtion and autolysis processes of yeast cellsunder the action of increasing alcoholconcentration; the content of these dead yeastcells becomes a valuable food for SMB. Untilthe stage of AF completion which correspondsto the complete metabolization (under 4 g/l) ofsugars by selected yeasts, SMB will attack ”thefirst course” from ” the nutritional menu” theyhave available and prefer the most (i.e. malicacid, the first substrate in the order ofpreference of metabolization), they willcontinue, if needed, with ”the second course”in case they did not have enough (i.e. with citricacid, the second preferred substrate) and onlyin exceptional cases (the selected yeast have notyet completed sugar metabolization and SMBshow one more viable cell density high enoughto require other nutritional substrates formetabolization) they turn to ”desert” (in thiscase, the remaining fermentable sugars non-metabolised by the selected yeast). Thistechnique requires a less stringent monitoringthan the previous one, in turn, it induces longerperiods of MLF (even with the use ofcompatible and appropriate nutrients).Triggering MLF through sequential inoculationwith SMB avoids the risk of Brettanomycesinstallation in the fermentative environmentthat would bring considerable prejudices on thesensory profile of the infested wine.


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» triggering mlf through staggered

inoculation. This technique of staggeredinoculation is applied after the completion ofAF of the must. This technique has less and lesssupporters, even though 10 –15 years ago it wasthe most appreciated technique, especially inthe Bourgogne region where it was regularlyapplied in dry white wines from Chardonnaykind. It still maintains its followers in the caseof producers to whom customers specificallyask for a Chardonnay whose sensory profile isdominated by a smell and taste typical of freshbutter („caractère beurré”).This asset of fresh butter is determined by theconcentration of diacetyl resulted after themetabolizaion of the citric acid by SMB whichusually occurs in the last third of the MLF.Since the technical equipment of laboratoriesof most wine cellars does not allow analyticalcontrol of citric acid metabolization, a puresensory evaluation is required (with a strongempirical character) of its metabolic state byassessing the intensity and the quality of thefresh butter asset which is determined by theamount of diacetyl resulting from themetabolization of this acid. In a wine which still contains fermentablesugars, the evaluation of citric acidmetabolization is almost impossible. For thisreason this staggered technique is appliedonly in the case of obtaining dry white winesfrom Chardonnay kind with a delightful assetof fresh butter.

Conclusions

Each of the four triggering and implementationtechniques of MLF in white wine making hasboth advantages and inconvenients. Dependingon the quality of the harvest, on the type ofdesired wine, on the technical equipment of thewine cellar and the laboratory, one of thetechniques presented above can be applied. Isuggest to interested parties the preliminary

completion of several comparativeexperiments on a pilot level (demijohns of 50l) or semi-industrial level (containers made ofstainless steel of 500 - 1000 l) at the beginningof the wine making process and only after anoptimum feedback the application of thechosen technique at an industrial level in thenext wine making process. The next issue of the magazine will beaddressed to the behavior and the comparisonof the same techniques of MLF triggering andimplementation in red wine making, with apresentation of protocols for managing SMBand compatible nutrients, but also with anapproach to problems regarding thecompletion of this process (on the evolutionof volatile acidity and highlighting the formsof lactic acid) in order to stabilize the winefrom a biological poin of view.

References

» Akin H., 2008 – „Evolution du pH pendantla fermentation alcoolique de moûts de raisins:modélisation et interprétation métabolique”,Thèse de Docteur de l’INP Toulouse.» Croitoru C., 2009 – „Treaty of oenologicalscience and engineering. Products ofdevelopment and maturation of wines”,Publisher AGIR.» Gerbaux V., Briffox C., Bou-Deleris M.,2008 - „Mise au point d’une nouvellebiomasse bactérienne pour l’ensemencementdirect des vins blancs”, Revue Françaised’Oenologie, 227, 6-11.» Gerbaux V., Briffox C., 2010 – „Faisabilitéet mise en oeuvre de l’ensemencementbactérien précoce pour les moûts deChardonnay”, Revue Française d’Oenologie,136, 13 – 15.


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Introduction

Malolactic fermentation (MLF) in red winemaking remains a topical issue for bothpractitioners and researchers despite thescientific progresses recorded in the last yearsregarding the performances of selectedmalolactic bacteria (SMB) strains. In red winemaking the most important aspect isrepresented by the stage in which FML istriggered in regard to the course of alcoholicfermentation (AF). Very recent research studies regarding theoptimization of MLF achievement in redwines with SMB and appropriate nutrientshave shown that this process is completed thefastest in the stage of sequential inoculation(immediately after the separation of the freerun wine from the pomace), then at the

precocious inoculation stage (when thedensity of grape pomace decreases around1020 g/l) and the co-inoculation stage (in thedebut stage of AF but after inoculation withselected yeast), followed by the tardyinoculation stage (in the free run wine afterthe complete sugar metabolization) and veryrare in the spontaneous cases (without SMBinoculation).

Materials and methods

i. the opportunity to accomplish mlf

with SmB

There is an impressive diversity of views in theinternational world of specialists in oenologyregarding the opportunity to accomplish MLFin red wines only under the action of a few


malolactic fermentation mlf:again in SpecialiStS’attention with

new technical-Scientific approacheS !aSpectS of the red wine making (2)

croitoru constantin


Abstract: Malolactic fermentation (MLF) in red wine making remains an actual topic for bothwine makers and wine researchers, despite the scientific progress recorded in recent years relatedto the performance of selected malolactic bacteria strains (SMB). Also in red wine making, themost important aspect is the stage for the MLF onset in comparison to the stage of alcoholicfermentation (AF). Very recent research studies regarding the improvement of the MLF performance in red wines withSMB and adequate nutriments have revealed that this process is carried out the most rapidly atthe stage of sequential inoculation (just after the separation of the free run wine from the grapepomace), and then at the stage of precocious inoculation (when the density of the grape pomacedecreases to around 1020 g/l) and the stage of co-inoculation (in the debut stage of AF but afterinoculation with selected yeasts), followed by the stage of tardy inoculation (in the free run wineafter the complete metabolization of sugars) and very rare in spontaneous cases (without SMBinoculation).

Key Words: red wine, malolactic fermentation, inoculation, Brettanomyces.

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strains of SMB belonging to an establishedgenus of Oenococcus oeni (O. oeni).» aspects of the traditionalist opinion.

Some specialists consider that eachviticultural area has the ability to make anatural selection of useful indigenous strainsof O. oeni existing in that area so that it is notnecessary the financial effort allocated forMLF accomplishment using commercialbiologicals of SMB since the indigenousstrains have a superior adaptation ability totechnological restrictive conditions of thefermentative environment specific to thosewines.Practical reality partially confirms thisattitude because in most cases the wine cellarsare unable to ensure a proper thermalcondition immediately after the completion ofthe wine preparation process, so that theprocess of MLF accomplishment is postponeduntil the following spring when thetemperature conditions often provide thetriggering of biological deacidificationprocess in the presence of indigenousmicroflora which certainly contains strains ofO. oeni and other less valuable bacterialspecies which inevitably contribute to theachievement of the process (and sometimesto the appearance of unpleasant smell or tasteas a result of the formation of biogenicamines, ethyl carbamate or as a result of a fewcompounds which seriously affect the sensoryprofile of the resulted wine).» aspects of the opinion of preliminary

comparative testing. There are plenty ofworldwide experts who are aware of theadvantages of using SMB to accomplish MLFbut are working in wine cellars that lack ofnecessary financial resources. The solutionlies in the experimentation at an industriallevel of the accomplishment of MLF withSMB on a specific wine lot (experimentalversion) however only through comparisonwith wine from the same lot at which MLF iscarried out with bacteria from the indigenous

microflora (control sample); for bothversions, it is recommended the comparisonof the MLF process durations and theconsequences that result from it, thecomparative sensory analysis, the evaluationof the costs of the experimental version andassessing the possibility of covering thesecosts through the price of wine as a result ofimproving its smell and taste characteristics.Comparing the same wine which resultedfrom different experiments but carried out inidentical technological conditions, theopportunity of MLF accomplishment usingbiologicals of SMB can be properlyevaluated.» aspects of vanguardist opinion. Anothergroup of experts (unfortunately the leastnumerous, but fortunately, rapidly growing!)managed to convince decision makers fromviticultural and wine making processessocieties in which they operate that thegeneralization of MLF accomplishment isopportune only with SMB at least for redwines from the “premium” and“superpremium” category. Companies that have generalized the MLFaccomplishment with SMB were under thenecessity of providing themselves with thenecessary technical equipment to achieve,in optimum circumstances, this biologicaldeacidification process; most importantinvestments were made in the climatemaintenance area of a few premisesintended for this technological purposewhich ensures appropriate heat treatment forthe triggering, the course and thecompletion of MLF. It is not coincidentalthat these companies have acquiredsignificant medals (the majority of gold !)at the most prestigious internationalcompetitions which they had participated inwith the wines of the above mentionedcategories and at which MLF wasaccomplished exclusively with SMB.


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ii. current techniques for SmB

utilization at mlf accomplishment

All these techniques differ from one anotherby the state in which the SMB suspension isadministered related to the progress stage ofAF which begins with the administration ofselected yeast suspension. Other aspects ofcurrent techniques of MLF accomplishmentwith SMB refers to the concept of co-inoculation compatible between selectedyeasts and SMB for some of these techniquesand of course at the danger of installation ofBrettanomyces yeasts for others.» Stages of bacterial inoculation. Sowingthe SMB can be done in different stages of AFrespectively before the AF onset (Croitoru C.,2005 as a bibliographical source, but it is notsubject to this article !), immediately after theAF onset (co-inoculation, simultaneously orshortly after the administration of selectedyeast suspension), during AF (precociousinoculation, in the last third of the AF whenthe density of the destemmed and crushedgrapes lowers to approx. 1020 g/l), in the finalstage of AF (sequential inoculation,immediately after the separation of the freerun wine from the pomace) or immediatelyafter completion of AF (tardy inoculation, inthe free run wine after the complete sugarmetabolization) (Croitoru C., 2009; BernierC., 2009) .» the concept of co-inoculation and

compatible cohabitation. For the first twotechniques involving a more prolongedcohabitation of selected yeasts with SMBwhich will have to take into account theconcept of compatible co-inoculation aswell; this new concept was introduced veryrecently by Renouf V., et. al. (2008); authorspresume that any couple of selected yeaststrain – SMB strain involves different typesof interactions that can be positive, neutralor inhibitory; considering the yeasts’activities which reflect the phenotypes of

each strain, the onset of MLF can befavoured or, on the contrary, slowed down;it is important to take into account thesecompatibilities between selected yeaststrains and SMB strains when applying theco-inoculation technique (or precociuosinoculation) in which yeast cells andbacteria have to cohabit; as for yeastsstrains, the ones which produce higherconcentrations of SO2 and medium-chainsaturated fatty acids will not berecommended for these techniques, a strainof yeast is choosen instead which producesthese unwanted compounds in extremelylow concentrations; the new concept is alsoapplicable to SMB strains of O. oeni genusthat have the ability to tolerate high osmoticpressures of the destemmed and crushedgrapes (those with very high sugarconcentrations) as well as higher AFtemperatures in red wine making; thesestrains will clearly be recommended for thetwo techniques mentioned above.» the danger of Brettanomyces yeast

installation. Numerous studies (Cauchy-Alvin B., 2005; Renouf V. et. al., 2008)showed the influence of a rapid MLF onseton cotaminated Brettanomyces yeast cells’density; the growth of contaminatedpopulation was stopped when the MLF onsetwith SMB takes place; unfortunately, oftenin wine production, late MLF onset when thecontamination level with Brettanomyces isalready high will cause difficulty in theaccomplishment of this biologicaldeacidification process as well as thepossibility of installation of some unwantedbacteria deviations. The situation iscomplicated when the wines were sulphitedin autumn with too large doses of SO2, sothat in spring, a late trigger and a slowprogress of MLF are noticed, creating theconditions of rapid development ofBrettanomyces yeasts.


20

iii. the co-inoculation technique

» generalities. The co-inoculation processconsists in the sowing of the destemmed andcrushed grapes with selected yeast and SMB,simultaneously or slightly delayed («simultaneous co-inoculation» or «quasi-simultaneous co-inoculation») byadministering SMB within 24 hours after theadministration of selected yeasts.An objective of a co-inoculation is to promoteSMB implantation in the fermentativeenvironment, taking into account that thedestemmed and crushed grapes, through itscomposition, ensures less stressful survivalconditions for these microorganisms than thewine because it does not contain alcohol, andthe available nutritional resources do notdetermine deficiencies in assimilable nitrogenand some vitamins; the destemmed andcrushed grapes environment in relation to thewine environment has the advantage that thebacterial sowing provides BMS a gradualacclimatization to the fermentativeenvironment with the increasing of its AFprocess.» recommended technological situations.

This technique of co-inoculation isrecommended especially when obtaining redwines which are intended to rapidcommercialization with high pH (> 3,4)bearing in mind the fact that an additionalawareness regarding their biologicalstabilization is compulsory.» eventual possible risks. The resultsobtained from co-inoculation experimentshave not confirmed the initial hypothesesregarding the risk of formation of somenoticeable amounts of acetic acid under theaction of SMB because the acetic acid ismainly produced when the SMB or otherbacterial strains have completed malic aciddegradation and began the citric acid.» the evolution of the biological agents. Inthe event of a co-inoculation, there is rapid

decrease in the bacterial population followedby its increase in the stage when the yeastcells reach the death phase. Experiments haveshown that bacterial cell survival is dependenton pH, and the survival rate increases oncethe pH increases. SMB leaven will beadministered only after the density of viablecells of inoculated selected yeast intended forAF reached the maximum level (this level isreached normally after 2 - 3 days fromsowing), so that yeast-bacteria interaction isavoided; after at least 48 hours from the onsetof AF, SMB leaven is added which isprepared according to the protocol presentedbelow.» technological parameters of reference.

Effective management of the co-inoculationtechnique requires several conditions for thefermentative environment necessary forbacterial development which relates to the useof destemmed and crushed grapes with atemperature over 17°C, a pH above 3,2 andtotal SO2 content below 50 mg/l. MLFmonitoring is recommended so that theprocess will be stopped after the malic acidhad been entirely metabolized or after a greatamount of citric acid had been metabolised orimmediately after the citric acid had beencompletely metabolised.» the necessity of rigorous analytical

monitoring. The choice of co-inoculationtechnique should take into account the must’sanalytical parameters (pH, sugarconcentration, SO2 content, assimilablenitrogen content). After the onset of AF,dosing the concentration in free SO2 isrequired and the stage at which thisconcentration is zero, the administration ofthe SMB suspension is possible; in addition,SMB implantation in the fermentativeenvironment should be controlled to ensurethat this process took place under optimumconditions using the microscopic method withepifluorescence applied after 24 hours frombacterial inoculation; it is also recommended


21

the regular monitoring of the volatile acidity,in addition to the monitoring of must’s densityand its L(-)malic acid content.» other specific technological aspects.

Through the co-inoculation technique, it isnoted that the SMB do not attack othersubstrates such as sugars, but consumes onlythe malic acid as the pH of the fermentativeenvironment is, in most cases, below thecritical limit of 3,6. When applying thistechnique, the latency phase of SMB isvirtually zero, the MLF duration isconsiderably shortened and the favorableimpact on the sensory profile of wines whichensures and provides its typicity isundeniable.It has been demonstrated that the bacterialinoculation suffers a significant loss of viablecell density due to the reduction of pHthrough an increase in acidity caused by theformation of succinic acid, lactic acid andpyruvic acid by the yeast cells as byproductsof their metabolism. Since the yeast cells ofsome strains have the ability to produce SO2but also to consume a large amount of thenutritional resources in this stage of AF itmeans that there are other technologicalreasons that do not recommend thisalternative technique.

iv. precocious inoculation technique

It refers to the SMB inoculation in the lastpart of the AF when the density of thedestemmed and crushed grapes drops toapprox. 1020 g/l. In regard to the co-inoculation technique, the precociousinoculation technique do not have largedifferences in the sense that it refers to thesame technological aspects except that theadministration of SMB suspension is delayedas opposed to the state of administration ofselected yeast suspension; it can be said thatthe precocious inoculation technique can alsobe called the sequential co-inoculation

technique which is a delayed co-inoculationof SMB in regard to the selected yeastinoculation.In his doctoral dissertation, Nehme N. (2008)studied the interaction between a selectedyeast strain of S. cerevisiae and a SMB strainof O. oeni regarding the impact of thisinteraction on the achievement of AF andMLF in sequential cultures (delayed co-inoculation) and mixed cultures(simultaneous or quasi-simultaneous co-inoculation); research has shown that in thecase of sequential cultures, the strain of S.cerevisiae exerted an inhibitory activity on thestrain of O. oeni through the resulted alcohol(a proportion of 75 % of inhibition) andthrough a peptidic fraction with the molecularmass ranging between 5 and 10 KDa (aproportion of 25 % of inhibition), while themetabolization level of the malic acid was upto 3,5 times higher for simultaneous co-inoculation as opposed to the sequential co-inoculation version; this research hadhighlighted the importance of choosing thecouple of selected yeasts strains - SMB strainfor maximum success of the application ofsimultaneous co-inoculation technique (orquasi-simultaneous) to achieve MLF.

v. Sequential inoculation technique

Sowing with SMB in the final stage of AFrequires their acclimatization to thefermentative environment which is hostile(low pH, high alcohol concentration) and lowin nutritional resources. Even if in the finalstage of AF the yeast cells die after theyrelease through passive release the aminoacids in the extracellular environment; thisrelease is followed later by a slow naturalprocess of autolysis which translates into anutrient enrichment of the wine; in somecases, the release of amino acids in theextracellular environment does not occurimmediately after the death of yeasts cells and


22

the autolysis process requires a longer periodof time (several months) in order to observe asignificant enrichment of the environment innutritional resources; for these reasons,several techniques of wine sowing withcommercial biologicals of SMB require theadministration of nutritional adjuvants (MLFactivators) to ensure the onset, course andcompletion of MLF; between these adjuvantsthe greatest effectiveness was shown by thoseproduced through thermo-enzymaticdegradation of the cell walls originated fromautolysed yeast cells to which it came uponthrough the experimental testing of severalsimilar products used integrally (extracts orautolysates of yeasts) or partially processed(milled cellular capsules or with variousstages of thermal degradation). The improvement of MLF kinetics can beinsured by administering the activator (e.g.FERMOPLUS MALOLACTIQUE product)directly in the wine to be sowed with SMB(the SMB choosing, see table 4 for of the firstpart of the article published in the previousissue in which advanced products belongingto BIOLACT range are presented) or byadding it in two stages (a part during theacclimatization phase of the SMB and theother part directly into the wine intended forsowing with SMB) according to the protocolof SMB reactivation presented below. As allactivators, the activators intended for MLFmust exercise a nutritional effect which willallow the bacterial biomass to increase(provided by yeasts extracts or components ofcell walls thermo-enzymatic degraded) and adetoxifying effect which will ensure cellsurvival (provided by an intake of cell wallsin lower stage of degradation).Numerous studies conducted in recent yearshave shown that the administration of theleaven from several selected strains of O. oeniwith a high affinity for the metabolization ofmalic acid and then citric acid can be carriedout in the final stage of the AF when up to 10

- 12 g/l of residual sugars are still to bemetabolised without any existing danger ofunwanted bacterial deviations.

vi. the tardy inoculation technique

The technique of delayed onset of MLF hasbeen associated with the state ofadministering the SMB directly in the winejust because of their compatibility. In thistechnique, regardless of the chosen SMBbiologicals, the manufacturers and thedistributors recommended the administrationof the rehydratable biological material onlyafter the completion of AF (when the reducingfermentable sugars content is very low, lessthan 2 g/l residual sugars).The followers of this technique argue that apremature inoculation with SMB when thewine is left with unfermented sugar content(the stage corresponding with the slow-downof AF as a result of the yeasts entering in thedecline phase) is prohibited and thereforeshould be avoided. In this undesirablesituation, the risk of accentuation of AF slow-down appears, which can cause a bacterialdeviation with serious consequences on thequality of the wine (the phenomenon of lactictart). At this stage, inoculated SMB arecharacterized by a very high density of viablecells, which coexist in the presence of a viablepopulation of yeasts with a cell densitysimilar to the bacteria as order of magnitude,but with a much lower vitality as opposed tothe vitality of bacterial cells.The acceleration of the death phase of yeastsdue to the presence of SMB in thefermentative environment has beenscientifically demonstrated and practicallyconfirmed; this biological phenomenon ofacceleration of the death phase of yeasts isdetermined by the hydrolysis of yeasts cellwalls under the action of the enzymeequipment available to SMB cells. Thebacterial inoculum multiplies taking


23

advantage of the presence of yeast autolysis(which is an excellent source of nutrition andexceptional growth) and it ferments theresidual sugars causing volatile acidity.Some SMB manufacturers and distributors tryto classify MLF after its completion with SMBusing direct inoculation or the preparation of areactivated leaven; the practice of wine makingprocess shows that the directly administeringof SMB into the wine without a priorreactivation was doomed to failure in mostcases, regardless of the strain’s performances.


A recently comparative study (seeexperimental versions presented at thebeginning of the article compared with thecontrol version unsown with SMB) regardingthe influence of the inoculation stage withSMB on the course MLF in the case of Merlotkind had monitored the evolution of kinetics offermentation and reported several analyticaldistinctions, microbiological distinctions andsensory distinctions at the resulted wines(Bernier C., 2009).In all experimental versions (including thecontrol version) the AF process was conductedin adequate circumstances and also performedwith the same strain of selected yeast. It wasfound that MLF has triggered the fastest in thesequential inoculation stage (immediately afterthe separation of the free run wine from thepomace), then in the precocious inoculationstage (when the density of the destemmed andcrushed grapes decreases around 1020 g/l) andin the co-inoculation stage (in the debut stageof AF but after the inoculation with selectedyeasts), followed by tardy inoculation stage (infree run wine after the complete metabolizationof sugars) and very rare in the case of controlversion (spontaneous MLF without inoculationwith SMB).Sequential sowing resulted MLF with the

shortest duration and the shortest periodbetween the end AF and the beginning ofMLF; SMB sown samples at the beginning ofAF (co-inoculation version) and in the lastthird of the AF (precocious inoculationversion) could not completely remove theperiod between the completion of AF and theonset of MLF but this period wasconsiderably reduced compared to the controlsample; SMB suspensions were implanted inthe fermentative environment to allexperimental versions so that the risk of yeastBrettanomyces contamination was limited to amaximum; no significant analyticaldifferences were found between the resultedwines; and in sensory level the highest-ratedwine was the wine obtained by sequentialinoculation namely the one which was thefastest in beginning and completing theprocess of MLF.

i. reactivation protocol of SmB

During acclimatization, BIOLACTACCLIMATEE selected bacteria are found inthe ideal pH and nutritional conditions so thatthey multiply faster than indigenous bacteriafrom the must or wine which continues toremain in very poor growing conditions. Inthe example below, the total volume of wine(must) intended for MLF is of 100 hl.Regardless of the volume of wine (must)intended for the treatment to achieve MLF,the preparing of the leaven from the samewine (must) is required which is a volume of1 % of the wine’s (must’s) volume intendedfor treatment. Therefore, in order to sow 100 hl, a volumeof 1 hl of leaven from the same wine (must)will be required. For other volumes of wine(must) intended for treatment to achieve MLF,the necessary quantities of SMB from theBIOLACT ACLIMATEE range andFERMOPLUS MALOLACTIQUE nutrientare shown in table 1.


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The preparation of the leaven implies goingthrough the following sequence oftechnological operations:1. The preparation of a recipient with acapacity of 1 hl, in which will be added 50liters of wine (must) and 50 liters of waterfree of chlorine, resulting a diluted suspensionwith a pH between 3,4 and 4.2. Administering of a dose of 500 g/hlFERMOPLUS MALOLACTIQUE (nutrientnecessary for growth and development ofbacterial cells which is a nutritionalsupplement based on autolysed yeast cellwalls degraded by heat and enzyme action) ina volume of 1 hl of diluted suspension,followed by homogenization .3. If possible, ensuring an optimumtemperature of 22 - 24 °C. Even a minimaltemperature of 20 °C can trigger MLF butwith a lower density of bacterial viable cells.4. Sowing the homogeneous suspension with afull dose of 100 g of BIOLACT ACLIMATEE(corresponding to a volume of 100 hl wine(must), namely 1 g of bacteria in 1 hl of wine

(must)) which should provide a density ofapproximately 2 x 10 7 viable cells/ml.5. Maintaining the optimum temperature (20- 22 ºC) for 24 hours for SMB acclimatizationat the wine restrictive environmentalconditions and for triggering the cellmultiplication. Periodic homogenization ofthe sown wine is recommended every 3 - 4hours with BIOLACT ACLIMATEE andenriched with FERMOPLUS MALOLACTIQUEnutrient. The assessment of homogeneityconsists in periodic visual examination of theleaven content in the recipient.6. A period of 48 hours at a temperature of 20- 22 ºC must be insured which will allow themetabolization of approx. 2/3 to 3/4 of theinitial malic acid content of the wine (must);this stage allows the progressive decrease ofpH and the avoidance of pH shock. Thesuspension in which the malic acid had beenmetabolized is represented by the reactivatedfinal leaven which is able to trigger, toundertake and to complete the MLF of thatwine (must) in an optimum period of time.


Table 1

the quantities of SmB from Biolact range

and fermopluS malolactiQue nutrient

needed to achieve mlf in different volumes of wine (must)

the volume of

wine (hl) intended for

treatment

the quantity (g) of

Biolact selected

bacteria

the quantity (kg) of

fermopluS

malolactiQue nutrient

50 50 0,250 + 1,5 = 1,750

60 50 0,300 +1,8 = 2,100

100 100 0,500 + 3,0 = 3,500

150 150 0,750 + 4,5 = 5,250

200 200 1 + 6 = 7

250 250 1,250 + 7,500 = 8,750

300 300 1,500 + 9 = 10,500

400 400 2 + 12 = 14

500 500 2,500 + 15 = 17,500

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Sowing the recipient with wine (must) withSMB leaven involves:7. Administering a dose of 30 g/hl ofFERMOPLUS MALOLACTIQUE adequatenutrient in the wine (must) from the recipientintended for achieving MLF with 24 hoursbefore adding the final SMB leaven.8. Mixing the final leaven with the wine(must) intended for MLF previously enrichedwith a dose of 30 g/hl of FERMOPLUSMALOLACTIQUE.

» the required amount of SmB. The doseof SMB remains constant (1g/hl) regardlessof the type of product that is usedused.Usually, packages are sold with a net contentof 25 g (for a volume of wine of 25 hl) or 250g (for a volume of wine of 250 hl), whichprovides the necessary quantity of SMB fordifferent volumes of wine (see table 1).

» the required amount of nutrient. Thenutrient is administered in 2 distinct phases:In the preparation of the reactivated leaven,when the required amount is calculated usingthe formula: Q1 = 5 g/hl x 1 % V x 100, inwhich: V = wine volume in hl

Example: Q1 = 5 g/hl x 1 % (100 hl) x 100 = 5 g/hl x 1hl x 100 = 5 g/hl x 100 hl = 500 g = 0,500 kg

» in the feeding course of the bacteria

during mlf, when the following formulaapplies: Q2 = 30 g/hl x V, in which: V = winevolume (hl) intended for treatment.

Example: Q2 = 30 g/hl x 100 hl = 3000 g = 3 kg

» total amount of the nutrient will be: Qt =Q1 + Q2 = 0,500 + 3 = 3,5 kg Note: The success of the wine’s MLF onsetincreases if the triggering of this process ofbiological deacidification occurs by the endof the alcoholic fermentation when itstemperature is of 20 - 22 °C, which provides

a rapid cell multiplication of SMB in both thepreparation of the final reactivated leaven andthe course of the malic acid metabolization.Attention must be paid to the pH of the winebecause a value below 3,2 should bemonitored! Other limiting technologicalfactors (alcoholic concentration greater than15 % vol. or a free and total SO2 contentabove the permissible limits of 10 mg/l and50 mg/l) usually occur very rare in thetriggering of MLF of the wine.

ii. managing the progress and the

completion of mlf with SmB

After the completion of malic acidmetabolization, SMB seeks other hydrocarbonresources to exercise their own metabolism.At this stage the increase of volatile acidityoccurs through the metabolization of the citricacid or the appearance of unwanted bacterialdeviations occurs due to the metabolization ofthe traces of sugars. If the period after MLF isextended, the wine becomes a favourableenvironment for the growth of severalpathogen germs of the Brettanomyces andPediococcus genus.The MLF process is completed when there areonly 0,1 to 0,3 g/l of malic acid, sometimeseven less. Apart from exceptional cases alsorequiring citric acid metabolization, rapidintervention is recommended for cessation ofthe activity of lactic bacteria. The citric acidtransformation can be appreciated or notthrough the formation of typical aromaticcompounds. The metabolization of traces ofresidual sugars with the formation of volatileacids is prohibited. A normal and pure MLFonly forms traces of volatile acids (+0,05 to+0,15 g/l), while in the case of lactic tart onsetthe volatile acidity can reach up to 0,4 g/l in afew days. Effective management of MLFcompletion requires a few additional analysis.» volatile acidity monitoring. Measurementof residual malic acid is not sufficient to


26

detect the presence of acetic bacteria in thefermentative environment so that the volatileacidity dosing is a complementary measurewhich allows rapid detection of severalpossible alterations caused by the presence ofthese bacteria. The determination of volatile acidity can bedone through the classical method(distillation) or through new methods(enzymatic method). In the case of thedetermination of abnormally high values(above 0,6 g/l in acetic acid) is necessary toidentify the microbial agent responsible forthis anomaly; the first stage is to observe,through a microscope, the bacteria present inthe wine using an optical microscope(magnification x 1000) in immersion; thisobservation often allows the identification ofbacteria and sometimes of the genuspossessing these specific characteristics; inthe second stage, a viable cell counting onspecific mediums can be done.

» highlighting the forms of lactic acid. Incase of lactic tart that occurs immediatelyafter MLF in the case of wines with high pH,the identification of the disease can be doneeasily by determining the stereochemicalnature of the lactic acid present in wine.Enzymatic dosage of the forms of lactic acidpresent in wine is very simple and very fast;a content of D(-) lactic acid greater than 300mg/l shows the occurance of unwantedbacterial deviations. Yeasts forme littleamount of lactic acid begining with themetabolization of hexoses of D(-) lactate,while the lactic bacteria only forms L(-)lactate based on L(-)malate during MLF. On the contrary, when the lactic bacteria usesugars, D(-)lactic results in the case of coccusand both forms of lactate, L(-) and D(-), inthe case of lactobacilli. Under theseconditions, the presence of a critical contentof 300 mg/l or more of D(-)lactic acid isrepresented by the characteristics of a

multiplication of lactic bacteria whichmetabolizes sugars from the environment.Therefore, the rapid elimination of lacticbacteria from the wine is recommendedimmediately after the completion of MLFwithout waiting to be noticed an increase ofthe volatile acidity which is another proof ofthe installation of unwanted bacterialdeviations.

» the influence of SmB usage on the

biological stability the of wines.

In general, the microbiological stability of awine is determined primarily by its pH value;the lower the pH value is the higher themicrobiological stability is because fewerbacterial genera and species can develop anda stronger antiseptic action of SO2 is signaledthrough a significant proportion of active SO2in the wine.The success off a sustainable biologicalstability of a wine is ensured by the limitingconditions of the development of lacticbacteria that refers to the acidity level, thealcohol concentration, a low storagetemperature and at the administration of arational dose of antiseptic.In view of the above, even if the pH valueincreases after MLF, the biological stabilityemphasizes; this phenomenon has nothingparadoxical because by the disappearance ofsugars and malic and citric acids, the easilymetabolised by the lactic bacteriasources of hydrocarbon disappear as well,simultaneously with the disappearance ofother nutritional resources as a result ofbacterial cell multiplication, to which it mightbe added the release by the lactic bacteria cellsin the wine of several toxins which inhibit themicrobial activity.The separation of the wine from the lacticbateria sediment along with its sulfitation arealso contributing to the biologicalstabilization.


27

Conclusions

The main advantages of MLF implementationwith SMB will be discussed below.

» faster mlf onset. Using SMB, chancesare high that MLF will trigger faster so thatthe duration of the process of red winemaking is reduced.

» mlf implementation in difficult wines.

A major advantage is the possibility of MLFimplementation in some difficult red wines(with low pH, high alcohol level and a contentof free/total SO2 exceeding 10/50 mg/l) as aresult of efficacious acclimatization of SMBto the fermentative environment and theelimination of the inconvenience related to arestrictive temperature (below 17 - 20 °C).

» limiting the risk of Brettanomyces yeasts

production. By inoculating SMB directly inthe destemmed and crushed grapes, whichensures their development immediately afterthe yeasts reach the death phase, the period oftime in which there could be a vacuum ofuseful microorganisms is limited to amaximum; this would allow the unwantedyeasts from the Brettanomyces genus to install.

» the reduction of biogenic amines

production. By using SMB, the biogenicamines production is reduced so that theresulted wines better conserve their fruitiness.

» improving the sensory profile and the

biological stability of wines. The reality ofwine production demonstrates that MLF withSMB is required for all red wines structuredand intended for aging because it gives theman added complexity, gracefulness androundness taste and ensures theirmicrobiological stability.

References

» Bernier C., 2009 – „Influence du momentd’inoculation en bactéries lactiques sur ledéroulement de la fermentationmalolactique”, Mémoire de fin d’étude pourle Diplôme National d’œnologie, Facultéd’œnologie Bordeaux 2, France.» Cauchy-Alvin B., 2005 - „Étude de laprédominance des bactéries sélectionnées surla flore contaminante de typeBrettanomyces”, Revue des Oenologues, 117,13 – 14.

» Croitoru C., 2005 - „Reducerea acidit`\iimusturilor ]i vinurilor. Metode ]i procedeefizice, fizico-chimice, chimice ]i biologice”,Publisher AGIR, Bucharest.

» Croitoru C., 2009 - „Tratat de }tiin\e ]iinginerie oenologic`. Produse de elaborare ]imaturare a vinurilor”, Publisher AGIR,Bucharest.

» Nehme N., 2008 - „Étude des interactionsentre Saccharomyces cerevisiae etOenococcus oeni: impact sur la réalisation dela fermentation malolactique en culturesséquentielles et mixtes”, Thèse de doctorat,École doctorale de mécanique énergetique,génie civil et procédés spécialité: génie desprocédés et environnement, Départament BioProcédés et systèms microbiens duLaboratoire de Génie Chimique, Toulouse.

» Renouf V., Gourraud C., Murat M. L., 2008– „Les différentes alternatives d’utilisationdes levains malolactiques. Partie 1/2: Aspectscinetiques et microbiens”, Revue desOenologues, 128, 39 – 43.


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Introduction

The experimantal plot has been established inChiscani, a locality placed 2km from Danubeand 15 km South of Braila City, all varietiesenjoying the same pedoclimatic conditions.Braila County is between the followinggeographical coordinates: 44º 44’ and 45º 30’N, 27º 04’ and 28º 10’ S. It is located insoutheast side of Romania, occupying thenorth-eastern Romanian Plain and the BigIsland of Braila, the largest embanked site ofthe Danube River.The relief is generally flat, the only „landaccident” being the rivers, and lakedepressions. Among the relief units, in theNorth side the county includes a part ofLower Siret River Meadow, towards the West

it includes small parts of Salcioara Plain andBuzaului Plain, and towards the East theLower Danube River Meadow.The relief of Braila County is of cumulative,sedimentary origin, the loess deposits havinga thickness ranging from 10 to 20 m. About75% of county’s surface is covered ofchernozem. Theese soils formed in thecontinental semiarid climate conditions andof loess deposits, on silt and sand, on theground of a steppe vegetation and surfacegroundwater sources.The most spread chernozem soils are:l Brown chernozem;l Chocolate chernozem;l Carbonate chernozem;l Leachade chernozem.

Braila County climate is temperatecontinental with shades of aridity.


Study of Some tomato varietieS originating in englandin the pedoclimatic condition of South-eaStern

romania, reSpectively Braila county

Galan C`t`lin, Bololoi Mihaela, Eremia Florentina


Abstract: The study has as the main target the testing of some Tomato varieties originating inEngland in the pedoclimatic condition South-Eastern Romania from Braila region. For this firststage of the project htey are not of special interest the technical economical parameters (costs,productivity, profit etc.), theese following to be the object of a different study. They have beenstudied six ,,cherry“ tomatos varieties. (Solanum lycopersicum var. Cerasiforme):

- Cherry White (C.W.)- Cherry Black (C.Bk.) - Cherry Brown Berry (C.Br.)- Cherry Gold Nugget (C.Gn.) - Cherry Cerise (C.Cr.)- Cherry Riesentraube (C.Rs.)

The biotechnical parameters of the cultivated varieties were monitorized along the wholevegetative period, ierespectively May 29th 2011, the planting date in open field, and until October22nd, the date of the experimental plot land clearing.

Key Words: ,,cherry“, tomatos, Solanum lycopersicum var. Cerasiforme.

29

Solar radiation has a relatively uniformdistribution, totaling between 122,5 and 125kcal/cm and about 2,200 hours of realsunburn, out of which 72% is in the periodApril-September.The high temperatures during the summerfrequently exceed 30-35ºC, being tipical forthe temperate continental climate. During thecold season, along about 110 days, it occursthe ground frost, out of which 80 days occursonly at night. Concerning the precipitation, the annualaverage is 465 l/m, most precipitation beingrecorded during May-August, and the least inautumn and winter.


Concerning the used culture materials andmethods, they have been covered the

following steps/technological links:»In order to shorten the acclimatizationperiod and to reduce the losses fromtransplanting, the six varieties have beenimported from England as seeds, theseedlings being produced in Romania (PictureNo. 01). The seeds supplier was PREMIERSEEDS, a British company.


Picture No. 01 – „Cherry” tomatos seedlingsready for planting in the field

Picture No. 03 – Giving Metiocarb granules before planting

Picture No. 02 – Land modeling

30

» Date of planting in pots was may 1st 2011,and the date of planting in the opened field wasmay 29th 2011, the efficiency being of 96 %. » The field where it has been established theexperimental plot, have been prepared inadvance by making a spring plowing at 30cm, and use of the combinator for raising andleveling. The land modeling have beenmanually performed by open 70 cm widefurrows (Picture No. 02). » Before planting in the field it was givenMetiocarb (2% a.i.) as granules in order tocontrol the fen cricket (Gryllotalpagryllotalpa) and other harmfull insects(Picture No. 03).Planting distances were the same for all

varieties, 70 cm between rows and 30 cmbetween plants on row (Picture No. 04),resulting a density of 47,619.0 plants/hectar.» The tomatoes grown in opened field, inintensive system, having a steam suportingsystem and flooding irrigation furrows(Picture No. 05).» The plants were periodically removed ofside-shoots but not pinched out. Five out ofthe six analyzed varieties shown an excelentforce of growth along the summer, reachingabout 1.60 m 60 days after the planting(Picture No. 06). The sixth variety, CherryGold Nugget (C.Gn.) shown a determinedgrowth..

» Weeds control have been performedmanually by hoeing between rows and bypulling weeds between plants on row. Theywere not used herbicides. They have been


Picture No. 04 – Planting distance betweenplants (30 cm)

Picture No. 05 – System of support polesfashioned from wood and bamboo crafts

Picture No. 06 – Plant height 60 days afterplanting

31

performed 4 hoeings every 15-20 days. » Concerning the irigations, thanks to thefrequent rains registered in the period may

30th Julyth 2011, the water consumption wasreduced, the irigations being intensified inAugust at a 3-4 days interval.


The biotechnologic parameters of thecultivated varieties where monitored alongthe whole vegetative period, respectively

may 29th 2011, date of planting in the openedfield, and until october 22nd 2011, clearingdate of the experimental plot. The performed observations can besynthetized as it follows: » The varieties C.Br., C.Cr., and C.W. showna special vigour of the shoots resulted fromside-shoots (Picture No. 07).

» Despite the large inflorescences, the varietyC.Bk. shown a high sensitivity to pollination,the yield obtained being low (Picture No. 08).» Although they have been periodically


Picture No. 07 – Lateral growth sprouts („side-shoots” type)

Picture No. 08 – Inflorescencesof the Cherry Black variety

(C.Bk.)

Picture No. 09 –Black spot on berries of Cherry Brown Berryvariety (C.Br.)

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Picture No. 10 – Berries of the Cherry Riesentraube variety (C.Rs.)

Picture No. 11 – Cherry Gold Nugget Variety (C.Gn.) when it reached the maximum yield.

Picture No. 12 – Berries of the Cherry White variety (C.W.) at the physiologic maturity

performed criptogamic treatments, the varietyC.Br. shown an high sensitivity to black spot- Alternaria spp. (Picture No. 09).» On reaching the physiological maturity, theberries of the variety C.Rs. shown unevenripening and thus a commercial derogatoryaspect too (Picture No. 10).

» The only variety with a detemined growth(C.Gn.), registered an average yield of 0.480kg/plant, plant height being of only 60-70 cm.It results an average yield per hectar of22,857.12 kg, level reached 70-75 days afterplanting (Picture No. 11).» Although the berries of the C.W. variety

33

shown uniformity and a special commercialaspect, it showed sensitivity to cracking,phenomenon amplified during the transport(Picture No. 12).

» The C.Cr. variety adapted excelent showing ahigh growth force, a long growing season, anda special quality of berries (Picture No. 13).

Conclusions

» All the analized varieties adapted well in thepedoclimatic condition of SE Romania;» The choosen culture system didn’t causespecial problem concerning the adaptabilityand productivity;» Five out the six varieties shown a specialgrowth vigour during the summer and a longvegetative period;» Concerning the yield quality, the obtainedresults with the varieties C.Bk. and C.Br. wereunsatisfactory;» At the C.Rs. variety the berries’ ripeninguniformity was poor;» The best results concerning the relationproductivity / quality were registered by thevarieties C.Cr., C.Gn., and C.W.

References

atanasiu n. – „Metode actuale de irigare înlegumicultur`”,. „Horticultura”, Magazine,No. 6, 2007;ciofu ruxandra, Nistor S., Popescu V.,Chilom Pelaghia, Apahidean S., Hogo] A.,Berar V., Lauer K.F., Atanasiu N. – „Tratatde legumicultur`”, Ceres Publishing House,Bucharest, 2004;dr`ghici elena – „Legumicultur`”, GranadaPublishing House, Bucharest 2002;dumitrescu m., Scurtu I., Stoian L., Gl`manGh., Costache M., Di\u D., Roman T., L`c`tu]V., R`doi V., Vlad C., Z`grean I. –„Producerea legumelor”, Artprint PublishingHouse, Bucharest, 1998;indrea d., Apahidean S., Apahidean Maria,Sima Rodica – „Cultura legumelor”, CeresPublishing House, Bucharest, 2007;oprea a., Galan C. – „Tehnologii generale ]ibiotehnologii speciale în agricultur`”,Course, I.S.B.N. 978–973–0–06230-4,Bucharest, 2009.popescu v., Popescu Angela – „Culturalegumelor în câmp ]i în solarii”, MASTPublishing House, Bucharest, 2003;http://www.premierseeds.co.za/vegetable-seeds/tomato.htmlhttp://stores.ebay.co.uk/Premier-Seeds-Direct/Tomatoes-/_i.html?_fsub=9717831


Picture No. 13 –Cherry Cerise Variety (C.Cr.)

34

Introduction

Rural tourism is based on three coordinates:space, people, products, because:

l space without human existence can notbe support cohabitation, a space withoutproducts may not meet all travel needs ofconsumers;l lack of space in people or products haveonly a small reception capacity;l products that have not the space andpeople that have only a fleeting existenceand can not ensure sustainabledevelopment locally.

The basic equation of rural tourism productis: accommodation on the farm = holiday invillage + leisure in rural areas.

At first rural tourism has become slowly butconsistently, especially in tourist markets ineconomically developed countries,

industrialized countries with a high degree ofurbanization. Thus in rural tourism isincreasingly popular and requested the peoplewho live and work in stressful conditions.

Practitioners of this tour can be found invarious areas of the world: in almost all ofEurope (European Community pays specialattention to projects and programs for tourismin rural areas), North America, Australia andeven Africa, standing out such flows tourismand concerns of practicing while tourismservices in the most unexpected areas.


a. motivational and strategic objectives

Rural tourism must be understood as apackage of socio-cultural, sports and activitiesspecific to the geographical environment,


proSpectS for development of rural touriSm in romania

Androne Roxana, Poparlan Alina Maria, Mihaila daniela Fanuta, Marica Mariana daniela


Abstract : In EU countries, rural tourism is not, in fact, a new phenomenon. Over time - this timebeing one in decades for most EU countries - tourist accommodation in rural areas was performedin a manner more or less spontaneous or organized. Instead, what is new is the size reached bythe expansion of tourism in rural phenomenon. This expansion can be explained on the one handby launching the development of rural regions and on the other hand, the diversification of thepractice of mass tourism. Therefore, rural tourism sign of Europe, one by one in the future localdevelopment policies. At first rural tourism has become slowly but consistently, especially intourist markets in economically developed countries, industrialized countries with a high degreeof urbanization. Thus in rural tourism is increasingly popular and requested the people who liveand work in stressful conditions.

Key Words: rural tourism, development of rural regions, development of rural tourism

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made disposal, they will take place inconditions of minimal investment and natureconservation. Climate and landscape,ethnography and folklore, ceramics, crafts,hunting and fishing fund, the fund viticulture,fruit growing, existing in rural areas, makeRomania to have multiple opportunities forrural tourism development.

– Strategic Objective

Analyzing the benefits derived from thepractice of this type of tourism, and currentsituation of Romanian villages it is necessaryto define Strategic Objective to be taken intoaccount in determining any strategy, be iteconomic or political, at any ruralcommunities.

The strategic objectives should aim to provideand protect the human factor, technicalfacilities and heritage conservation, groupedinto three strands, namely:

lStop the migration of the rural populationto urban and stimulate recovery eel, eelleast partially, the population of urbanareas to rural areas;l Ensuring the living conditions in ruralareas and of civilization, stimulating activepopulation stability in rural areas;l Conservation and environmentalprotection areas - factor of attraction oflocal population and foreigners to ruralareas.

Tourism in rural areas, complementary toother forms of tourism, contribute tosustaining the necessary conditions fortransformation tourist tourism tour circuit tostay.

This can and should become the tourist areaof residence, to spend their holidaydestination all year round, because it respondsnot only motivation and knowledge ofcultural tourism, but also other modern

requirements: to leisure in nature, whichotherwise found in other countries(investigations conducted among visitors tohistorical monuments show that attention isnot only the monuments but also on theirenvironment).

Motivation can be expressed as a convincingstatement of Dr. Andre Lwoff, Nobel laureate:“you’re a european metropolis lived in

noisy environments of continental explain

that here in the upper country, as you call

it ., those in romania, and live fulfilling

feeling to know peace... the beauty and

ambiance visited monasteries, quiet

environment, this is what struck me was

going to set there.“

B. factors that favor the development of

rural tourism in romania

As the output of the current economic crisisand rising income, demand will improvetourism in our country, but it is possible toorient toward less traditional tourism, whichwill remain expensive, and may in a greaterextent to rural tourism, which is cheaper.

On the possible direction would contributeand specific factors:

4 mountain, hill and many other areasoffer breathtaking landscapes;4 family agricultural offer natural food,prepared according to traditional recipes.4 Romanian population is largely apopulation who household still keepsparents or possibly nostalgia grandparents,who grew up in;

Preparation of this type of tourist offerrequires a series of efforts and actions atvarious levels to increase satisfaction andattracting demand:


36

Arrangement of the agro-tourist farms

Development of infrastructure localCreating structures organizers

c. perspective of rural tourism

development in lowland areas, hills and

mountains

Alecu Ruso noticed well, or poetic sense, theharmony of natural architecture of Romania,said that when our country is one of the mostblessed by God.

a) lowland area - includes land in S, Vand E of Romania. These regions are areasof greatest agricultural importance,benefits about 3 million hectaresdeveloped for irrigation and drainageworks. Network structure is the typelocality collected, with a density of 2 to 6.8km at 100 locations.

In terms of rural tourism plain area can counton a particular tourist vocation in the next 5-10 years, which can be determined by theexistence in the area of object of touristinterest such as lakes, historical sites, literally,forests, etc..

b) the area of the hill - is defined interms of geography, as the area thatcomprises the hilly terrain that is dominant

with altitudes between 300-600 m. aidrural settlements have a density of 0.4 to100 km villages and settlements networkis dominated by the diffuse type, the resultof natural conditions. This area lends itselfto a multifunctional agriculture andstructures land can be based on smallfamily farms.

In terms of rural tourism is one such areavocation because of higher overall variedlandscape and climate favorable.

c) mountain area - comprising landtypically located at over 600 m altitude.This area is characterized by a agrarianstructure of the plot, the average area perhousehold farm is 2.5 - 3 hectares.

In terms of rural tourism the area has alwaysexerted a powerful attraction, especially whendue to urbanization today and pollution cleannature of a human need is growing.

d. conditions for development of rural

tourism in romania

l In our country the forms of tourism in ruralareas can get a great extent.

ñ urban population - represents 55% ofthe total population, as such it representsan important potential for rural tourismdemand if it combined with factorsconsiderations that favor development ofrural tourism in Romania;

ñ free time - there. Vacations are aboutone month per year and work week is onlyfive days, after 1989. Therefore the activepopulation employed would have to spendtime both weekend and longer vacationsaway from home, which sometimes feelsthe need to escape;


37

ñ real incomes - but is more restrictivefactor that hinder eel, even block domestictourism demand. As long as the vastmajority of urban population is forced tospend 60-80% of their income for food, wehope to remain weak for travel money.Scourge of inflation which greatly reducedthe real incomes of population andpurchasing power has declined from ‘89(though not before this time the situationwas not very good) made internationaltourism demand to know a strong rebound.Foreign tourism demand is hindered byeconomic, social and political. The qualityof services we are foreigners in Romaniaare still below European standards. Eventhough in terms of national currencyexchange rate and prices compared tointernational markets stabilized, touristscould be motivated to come to Romania,however, the fragility of political andsocial security and personal failure createsa braking reaction flow of foreign touriststo Romania;

ñ relative inelasticity of classic

tourism - is manifested in Romania as alimiting factor, more so with this type oftourism as economic crisis lives of thetransition. Given inflation and theviability of the tourism needs, prices andtariffs have become prohibitive for masstourism potential, without the services tobe improved and modernized inproportion;

ñ motorization families - is a growingphenomenon. Paradoxically, although realincomes have decreased the number ofcars and construction of houses hasincreased both the city and the countryside.Engines were in full progress, createsconditions for potential tourists travelingin rural areas. Do not forget that ruraltourism is largely a form of family travel

and car travel is essential for family travel,especially where no other means oftransportation.


Rural tourism development must integrateboth the overall strategy for tourismdevelopment in the overall strategy as well associo-economic development of this area. Forthis purpose it is necessary to know inadvance of items:

l assessment of all categories of economicresources, financial and social place;l defining the directions and guidelinesfor economic growth in the area;l assessment of tourism developmentpossibilities and limits.

Arranging travel the countryside is a complexaction covering a wide problems includingsolutions to stimulate tourist traffic and theintegration of tourism in rural space economy,and methods of selection and delimitation ofareas, principles of operation.

– types of visitor facilities

Localizations are influenced by the size oftourism and tourist resources spreadthroughout the country. So meet:

a) locations related to the existence of asingle object or tourist item of great value,for example, Neamt Fortress, Bear Lakefrom Sovata. In these cases, arrangementsare summary dimensions are closelycorrelated with the index of theattractiveness of that objective. b) locations integrated into a set ofconditions that gives a certain specificityof geographical areas with a specificcharacter, for example, with the


38

monuments of art, architecture. In thesecases, more complex facilities are takingthe form of centers or tourist resorts.c) specific locations of tourist areasrelatively homogeneous, with a wide rangeof stretch, without a certain feature. Inthese cases, facilities are more numerousand can be simple or complex, and can beplaced anywhere watching brief is arelationship between resources andequipment located. In this case thedecision is determined decisivelydevelopment of tourism demand.

Besides these types of visitor facilities,reports on tourism planning experience of thesix regions of the World TourismOrganization, offers and other travel planningsolutions depending on the nature of theterritory in geographical space:

– Planning spa sites literally;– Land sites mountain;– Land of historical and archaeologicalsites;– Rural land;– Planning spa resorts, etc..

!!! site = French site, landscape meaning,position, region, geographical configurationof their own localities, and is dedicated toaccept broader theory and practice in spatialplanning tourism in many countries.

– Selection and delineation of tourist

areas

Depending on the delimitation of touristresources and determining on this basis, thedominant profile of each area, is a milestonein arranging travel to the territory. Visitorfacilities are consistent with each area takinginto account the potential surface to bedecorated, climate, vegetation, culturalartifacts, labor resources and demographic

situation, needs to stimulate economic andsocial progress of the territory

When the decision on the orientation of atourist development area, specify thedestination to be assigned to it must be suchas to ensure a customer demand morediverse.

In this regard, planning policy must take intoaccount the kind of tourism that has the bestchance of development in the area and toadapt their focus to specific typology. Inaddition some development principles andrequirements of their regions, the selection oftourist areas and are considering a number ofspecific criteria, namely:

l natural conditions (elements of touristattraction, the attraction function, typologyfacilities);l cultural and historical richness

(archaeological, historical monuments,traditions);l socio-economic conditions (demographyplanning, standard of living);l infrastructure (transport and ways andmeans of access, water sources, energy);l general equipment for culture and

recreation (means and forms of receptionand accommodation, food units);l legislation and regulations (have a rolein stimulating or restrictive depending onthe legal status of any perimeter of eachdevelopment must comply withregulations on environmental protection,construction rules).

unit types that will develop tourism in thetourist zone depend on the nature of theterritory, specific resources and features,density and their value.

Landmark tourist attraction is an interestingenough to motivate tourists travel to it(natural monument, archaeological sites, etc.).


39

In the context of rural tourism, the basic unitof activity tourist farms and agro tourismpensions. They provide the necessary servicesin the area, especially the reception,accommodation and meals.

Conclusions

1. a promising start -in Romania for overtwo decades by promoting rural settlementsethno-folk values , cultural and sceniclandscape as tourist villages was established,official rural tourism .

After 1989 the first initiatives began toappear, first at the level of animation‘s heartymountain tourism, such as Radu Rey, then themountain areas in Romania Commission andthe Ministry of Tourism. Thus the law waspassed approving Government Ordinance No.62 of 1994 on the establishment of facilitiesfor the development of rural tourism in themountains, the Danube Delta and Black Sea.They also established ANTREC. In manyrural villages appeared to profile companiesthat made accommodations in private homes,offering natural products and activitiesspecific to ice.

Tourists from Germany, USA, France, GreatBritain and elsewhere. The president said thatthe company is a member ANTRECEuropean Federation of Rural Tourism in theAmerican Society of Travel Agents of theU.S. Society of Ecological Tourism in theU.S., etc..

Romania has great possibilities ofdevelopment of tourism, its practice is notonly possible but also very necessary in thisstage.

Income from this activity will contributesubstantially to raising the civilization of theRomanian village and its inhabitants, the

hospitality so characteristic of the Romans,could change the image of Romania in theworld.

By the conclusion of partnership agreementswith national, regional or local profile, itwould ease access to EU funds for Romania’srural tourism, which Romania, currently cannot receive more than coincidence, since thereis still a member of the EU. It is alsonecessary to form a group of experts able toprovide technical assistance and training tokeep the local power bosses and owners ofstructures on the problems of rural tourismmarketing, planning and interior partitions ofthe housing and training and food servicesclassification and quality standards, cateringand dining rules, rules of hygiene andecology, behavior in dealing with tourists,information system.

2. rural tourism and socio-economic

development of the romanian village -Romanian village - rural tourism contributesto the economic life of the village by these

– Opportunity of a policy of its long-termdevelopment, in close liaison with othersector policies, agriculture, infrastructure,environmental protection;– Can become a support for newbusinesses and jobs;– Encouraging local traditional activities;– Dynamic factor in the process ofincreasing the quality of life in rural areas,etc..

3. Support and internal resources - formany years, countries like France, Austria,Germany turned to a new form of tourism

– Eco-tourism areas. Why not Romania?The wooden churches of Maramures, themonasteries of Moldova, crafts and thehospitality villages and Roman, houses withporch - these are universal values and


40

arguments for tourist attraction. In order topromote rural tourism, the professionalinform those who want to turn to this newfield, was established since 1994 ANTREC.

4. turismul areas (a lifestyle) - one of thetrends development of tourism industry isreturning to nature, already manifested in allcivilized countries where the current post-industries has created the need for withdrawal(for a few days) of metropolis. And for thehuge tourism potential of our country,

Rural tourism is one of the most attractivetourist offers, and this from a multipleperspective: on one hand the novel or foreigntourists, which will require increasingly moreintense this type of tourist service and on theother hand, villagers, who will use all hisenergy and skill to integrate into what willbecome, after efforts associations, a networkcovering the whole country.

Ministry of Tourism is heavily involved inrural tourism development. Importantinitiatives extends beyond legal forms that arerequired to prepare. The EU’s PHAREprogram was attached to a componentsupporting rural tourism development.

Joint development efforts of rural tourismwill certainly be rewarded in the near future.What concerns us all is, in fact, achieving ourcountry’s tourist vocation, whichunanimously recognized, is the mostbeautiful country in the Europeangeographical area. If this will be done, wehave a standard of living in developedcountries agree with the eel, as twenty-firstcentury tourism industry will be.

References

» Balaure V., 2006 - „Promotionaltechniques”, support the course, UniversityBioterra;

» Baciuman I., 2005 – „The economy ofrural tourism and agrotourism”, Alba Iulia;

» Bran Florina, 2006 – „Rural Tourism -European model”, Ed Economics;

» Bucianu C., 1999 – „The economy of ruraltourism and agrotourism”, Alba Iulia;

» Costea R., 2002 - „Marketing travelagents in the market economy”, NationalInstitute for tourism training andmanagemet, Ministry of Tourism;

» Deroni M., 2007 – „The situation of ruraltourism in Europe”, department MFAForeign Relations, Bucharest;

» Gavrilescu C., 2009 – „Marketing”,Bucharest;

» Radu E., 2005 – „Service Management”,Cantemir Christian University;

» Nistoreanu P., 2006 - „Rural tourism - asmall business with great prospects”, Ed.ASE;

» Nistoreanu P., 1999 – „Rural Tourism - asmall business with great prospects”,Didactic and Pedagogic Publishing House,Bucharest.


41

Introduction

According to FAO food security is “alwaysguarantee each individual at any place or timeof access to sufficient and healthy food toallow him to have a satisfactory diet for ahealthy and active life.”Food security isinfluenced by four groups of factors:

Social-economic and political environment

– Food sector performance– Social protection– Health and hygiene

The main bodies involved in monitoringglobal security are: FAO, EC, USDA. Inrecent years, says there is food insecurityproblems in 86 countries, 43 from Africa, 24from Asia, 9 from Latin America andCaribbean, 7 from Oceania, three in Europe.In 2004, 35 countries have received aidemergency because of the food crisis. Thecauses were: military and civil conflict, postconflict situations, refugees, economic zonesand climate problems. Romania, as is known,is aimed at joining the European Union on 1

January 2007. To achieve this goal, Romaniamust meet several conditions among which isthe adoption and implementation of theCommunity agreement , which includesbesides legislative harmonization andadoption of European standards in all areas,especially those that relate to health andpeople safe.

Romanian companies may have two options:either to adapt to new conditions, progressand prosper or lose their market position andare sad it bankrupt by misalignment to thenew European standards.

During 2-4.06.2005 Chamber of Commerceand Industry organized by the CenterTimisoara EUROINFO a Europeanconference in the project “Food Safety, anatural consequence of productresponsibility”, funded by the EuropeanCommission’s PHARE program.

At the conference representatives haveactively participated in 5 countries: Poland,Estonia, Hungary, Bulgaria, Italy. Theconference presented the followingdocuments:


the concept of food Security in europe

Stoica Valeri2, Atudosiei nicole Livia1, Androne Roxana1, Eremia Florentina1

1Bioterra University of Bucharest2Academy of Agricultural Sciences and Forestry of Bucharest

Abstract : European Nutrition was founded in 1967 on the foundations of internationalconventions. In an initial phase was conditioned by a community management products. Food aidmay thus vary from year to year depending on the reserves of European agriculture. Much later,after a deal between EU and ACP countries, technical and financial aid were donated to thebeneficiary countries in order to gain sufficient nutrition. Until then reflect the commission‘sdesign evolved from a food aid as a tool, among others, that contribute to improved food security,access to food in concrete terms.

Key Words: food security, management products, satisfactory diet.

42

â management systems;â HACCP principles and documents;â food safety management system;â Presentation of ISO 22000 standard forcertifying food safety system.


Application of HACCP system in all unitsthat make up the food chain is required underRomanian law, which requires that allcompanies in the food and catering motorscomply with the principles of this system.Standards contribute to implementation ofHACCP, developed by the CODEXALIMENTARIUS Commission and adoptedin Romania are:

Sr 13462-1:2001 agro-food-hygiene:

general principles: This standard followsthe road of food production from primarystage to the final, it reaches the consumer andestablish the necessary hygiene conditions forproducing a safe and adequate foodconsumption;

Sr 13462-2:2002 agro-food-hygiene:

the hazard analysis and critical control

point (haccp) and its implementation

guide. The standard establishes principles forrisk analysis and critical control points at playand how to identify these specific risks andcontrol measures in order to ensure foodsafety;

Sr 13462-3:2002-hygiene foodstuffs.

principles for the establishment of

microbiological criteria for foods at any

point in the food chain from primary

production to final consumption. Thisstandard specifies the principles ofestablishment and application of

microbiological criteria for foods, ensuringfood security mainly through source control,process control and application of appropriatehygiene practices during production,processing (including labeling), handling,distribution, storage, marketing.

Application of the standards mentioned aboveprovide compelling advantages:

4 Maintaining a market position;4 a safe sanitary quality;4 a reduction in scrap and customercomplaints;4 Increased customer confidence in acompany;4 Improving a company’s image.

Food safety management system integrates:

4 HACCP principles;4 Applying the steps described by theCODEX ALIMENTARIUS Commission;4 Combining dynamics with thenecessary prerequisites programs tocontrol and reduce any risks to acceptablelevels of food safety for the final productdelivered in the next stage of the foodchain;4 Hazard analysis that is essential foreffective food safety management systemas organized knowledge required to designan effective combination of controlmeasures.

Safety dietary essential qualities of a food, afood is safe when the consumer does not alterand endanger the health of the body.

food safety management system is

presented in iSo 22000 internationalstandard that specifies requirements for a foodsafety management system in a food chain, inwhich an organization:

â Must demonstrate the ability to controlfood safety hazards in order to provide safe


43

end products that meet food safetyrequirements agreed by consumers and theregulator;â Aims customer satisfaction througheffective control of food safety hazards,including processes to update the system.

The international standard ISO 22000specifies the requirements that allow anorganization:

â a plan, design, implement, manage,maintain and update a food safetymanagement systemwill evaluate and assess customerrequirements and demonstrate compliancewith mutually agreed customerrequirements related to food safety;â a demonstrate effective communicationwith customers and other stakeholdersalong the food chain;â will demonstrate compliance withapplicable requirements of the regulatoryauthority with respect to food safety;â will ensure that they meet establishedfood safety policies;â will demonstrate that under otherstakeholders;â to request a registration system or itscertification of food safety management byan external organization.

International standard ISO 22000 onlyconsider concerns about food safety issuesbut not discouraged by the organization andintegration of other aspects of themanagement system elements such asquality, in general and / or environmentalprotection.

Organizations that wish to take into accountsuch matters can do by implementing theinternational standard EN ISO 22000 withISO 9001: 2001 and / or SR EN ISO14001:2005.

Food safety and standards issues in the fieldhave been widely presented and discussed atthe European Conference which provided theoccasion attended by economic agents tounderstand the priorities for developing a safebusiness based on the new marketrequirements.


a. detection of genetically modified

organisms and foods derived there from

Today, varieties of soybean, cotton, corn andcanola are genetically modified products on25% of cultivated areas in the world. Genesin these plants were introduced to conferresistance to certain herbicides or toxicproducts used to control specific insects. Inthe United States produced nearly two-thirdsof the yields obtained by bio-engineeringglobally. Other countries where crops withgenetic alterations are major quantities are:Argentina, Canada, Brazil, China and SouthAfrica.

ISO standards to be developed will berelevant globally and will facilitateinternational trade and services by providinga consistent international approach todetection and analysis of foods derived fromgenetic organisms (GMOs). This will allowthe storage of seed industry, seed producers,grain, food manufacturers and analysislaboratories to meet the labeling provisionsthat have been developed by many countriesaround the world or who are currently in theplanning stage, being control tool used byfood control authorities in conformity withlegal requirements.


44

B. labeling provisions

When the United States exported for the firsttime in 1996, in Europe, soy genetic changes,the European Union in response to publicdebate, introduced obligatory labeling forfoods derived from organisms with geneticchanges. In April 2004 new legislation tookeffect that extended the labeling provisions.Other governments, following the example ofthe European Union have developed labelingprovisions for food derived from geneticallymodified food. Whenever food labelingapproaches with different genetic changesgreatly from one country to another.

c. detection of genetic changes in food

Understand range of DNA technology, whichled to obtaining genetic organisms, allowedthe development of detection methods andprecise carecteristice. To determine if a foodhas been produced from a GMO, the methodchosen was on search sites modified DNAsegments and / or new proteins resulting fromgenetic modification. The following figureschematically illustrates the detection ofmaterials derived from organisms withgenetic modifications in food. The startingpoint is an adequate sampling to obtain arepresentative sample of the productinvestigated. The next step is extraction ofprotein or DNA in the sample studied. Proteinextracts are taken at random. Extracted DNAanalysis is subject to RPL - polymerase chainreaction.

Conclusions

Romanian producers are forced to obtainquality certificates from international bodies.Certificates issued by RENAR nationallaboratory accreditation body for certification

of Romanian food, are not recognized by theEuropean Union.

Testing laboratories, certification bodies andsystems management product certificationbodies can not issue valid certificates abroad.European Union withdrew its accreditation afew months ago Accreditation AssociationRomania - RENAR national laboratoryaccreditation body for testing / analysis,metrological calibration, inspection bodiesand certification (quality management system/ environment, HACCP, product / servicesand personnel).

reaSonS. The suspension was decided,because there were non-compliance. Staffhave the competence and deficiencies thathave not provided evidence to ensuretraceability of measurement results. “After allconverge to the staff. It is an acute need toRENAR staff, “said association president,Prof. Fanel Iacobescu, which is part of thenew management team named after findingproblems by international bodies.

lack of SanctionS. Problems wouldnot have occurred if the certification bodieswould have had a duty under the rules or ifRENAR would be sanctioned in the wrong,admit president. The new president does notrecognize that sanctions have been appliedRENAR them Romanian certification bodieshave not made the right job. It is almostcertain that the sanctions now come also. “Wewill decide in two weeks for the areasrequested the assessment. It can be done inone year from the date of request. Good forentry into the EU we have many areas,“Iacobescu estimates, which estimates thatinternational organizations that providereliable current RENAR part management.

aBroad. Until then, those seekinginternationally recognized certificates areforced to go under another umbrella and be


45

certified or accredited to an internationalbody. “ Unfortunately, the letter of credit wassuspended creditor monitoring. Customers areconfused between a suspension ofinternational recognition of the limited termof the right body to accredit certificationbodies in Romania. Right to be a full memberhere, in Romania, took not one, “said MarianaIonescu, general manager of SIMTEX -Certification Body. Now manager SIMTEXOC desire is to accredit one of the mostpowerful bodies in the EU.

local brake eXportS “Most Romanianexporters have made accreditationcertification to institutions abroad, preciselybecause there was a lack of confidence inRomanian bodies certification, believes JohnCezar Coraci, president of UGIR 1903,member organization in the College RENARbeneficiaries. He estimates that non-recognition is a major European brakeRomanian export as these problems isparticularly important if you want to balancethe trade balance, “says John Cezar Coraci,president UGIR 1903, an organizationbringing together major employers in theindustry.

what are quality standards ?

ruleS. Alcohol manufacturers are requiredto have ISO 9001 and HACCP.

iSo 9001 - Quality management systems - aset of rules and procedures that anorganization always respect them.“Unfortunately, the confusion at the consumerlevel. The fact that a manufacturer has notcertified management system tells me, as aconsumer, that products are of quality. Hesays that control of product manufacturingand keep them well under control and that theproduct is always the same. This is why somemanufacturers who know the confusion and

write the note forces ISO 9001 certifiedproducts. It is not very fair for the product isnot certified, “says general manager RENARCristian Dorin Nichita PhD.

iSo 14001 - This standard refers to themanagement of environmental issues,specifically what makes the organization tominimize adverse environmental effectscaused by its activities and to continuouslyimprove environmental results.

haccp management SyStem - It isa system that identifies, evaluates and controlthe risks relevant to food safety.

accreditation Speed againSt time -directors will certificates quickly.Now the fashion is the food. Alcohol-producing companies are obliged by law toobtain the tax stamp and have ISO 9001certification risk. Another legislativeregulations require food companies to haveimplemented a system and risk analysis andcritical control points.

2000 euro.

Both ISO 9001 accreditation costs for a formwith 40 employees, the cost is around 1500euros, plus EUR 500 for annual monitoring.Fees for certification are based on the numberof auditors audit the number of days andnumber of personnel in the organization to beaudited. There is no fixed price once.

top iSo 9001

The biggest fans of quality managementsystems and of the environment areEuropeans. They have 48.76% and 43.96% ofthe certificates. U.S. has only 37,285, evenless than Spain by nearly 40,000 certificates.Great amateur certification is and China.

It has, moreover, ranking in the country with132 926 certificates issued.


46


the world

China 132, 926Italy 84 ,485United Kingdom 50 ,884Japan 48,989Spain 40, 972

in europe

Romania 5183Bulgaria 1685Czech Republic 10 ,781Hungary 10, 207Poland 5753Slovakia 2008USA 37, 285

References

» Ardelean, D. 2002 - „Probleme desecuritate alimentar` ]i integrarea Românieiîn Uniunea European`”, Editura ASE,Bucure]ti;

» Banu, C. si colab. 2002 - „Manualulinginerului de industrie alimentar`”, vol. I ]iII, Editura Tehnic`, Bucure]ti;

» Basdekis, J. C. 1999 - „Ghid de dietetic`”,Editura Polirom, Ia]i;

» Diaconescu, I. 1998 - „Merceologiealimentar`”, Editura Eficient, Bucure]ti;

» Diaconescu, I. 2002 - „Func\iile produsuluialimentar”, Editura ASE, Bucure]ti;

» Diaconescu, I. 2007 - „Merceologiealimentar`”, Editura Qlassrom, Bucure]ti,2004;

» Viezuin`, V. 2000 - „Securitateaalimentar` în lumina exigen\elor actuale”,Tribuna Economic`, nr. 2/12 ian;

» Zamfir, I. 2000 - „Protec\ia consumatorilorde la A la Z”, Editura Premier, Ploie]ti;

» Zamfir, I. 2002 - „Protec\ia consumatorilorpe în\elesul tuturor”, Editura Dacia AS,Ploie]ti;

*** FAO, 2004 - „The State of FoodInsecurity in the World 2004”, FAO, Rome;*** http://apps.fao.org *** http://departments.oxy.edu/uepi.schoolfoodschecklist.htm

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