Integrated research in Oenology

Full text

CONTRIBUTIONS to SCIENCE, 1(3): 365-369 (2000)
Institut d’Estudis Catalans, Barcelona
The creation of the Oenology degree at the University of Tar-
ragona meant a considerable challenge for the staff in-
volved. The initial period of consolidation of the teaching
function was followed by research development, the other
high priority of the Faculty of Oenology. The integration of
personnel from different scientific and technological back-
grounds allowed an excellent complementarity and synergy
for the development of competitive research projects. Cur-
rently, more than 40 researchers are involved in oenological
scientific and tecnological research, and these researchers
make up the Oenology Unit, a part of the Centre de Referèn-
cia en Tecnologia dels Aliments (Reference Centre for Food
Technology), a virtual research institute created by the Gen-
eralitat of Catalonia.
Origin and development
The School of Oenology, created in 1988, meant the develop-
ment of an experience in teaching and research in the disci-
pline of oenology. The need for quality teaching in a field that
was not yet recognised by an official title in Spain was a chal-
lenge for the group of researchers in the Faculty of Chemistry
who were involved. Initially, the focus was on teaching and a
curriculum development for oenologists that would meet the
needs of the rapidly growing wine sector. Although this initial
goal was achieved, the final accreditation of the 1996 Bac-
calaureate of Science (BSc), in Oenology changed the old
School of Oenology into the present Faculty of Oenology.
The academic restructuring and teaching delivery re-
quired an additional effort in fundamental oenological re-
search. Thus, the personnel involved with the Oenology
School during this time had to develop research pro-
grammes within the wine sector. This required strong scien-
tific disciplinary interaction among the scientists as they
faced different research projects. Also, new winery facilities,
experimental and sensory evaluation laboratories, were es-
tablished to be used both for the formation of oenology stu-
dents and for research.
In 1994, the Generalitat of Catalonia decided to develop
the «Reference Centres for Research» within their «Re-
search Plan». The «Reference Centres for Research» were
designed as virtual centres which aggregated some already
existing facilities in public research institutions. One of those
Reference Centres was the Centre de Referència en Tec-
nologia dels Aliments (CeRTA, Reference Centre for Food
Technology), which had 7 Research Units. Among these
was the Oenology Unit, grouping the researchers involved in
developing the different research activities globally related
to wine. This global approach allowed the development of
research streams ranging from vineyard training and grape
production, to wine fermentation and development, quality
improvement factors which related to the health and nutri-
tional aspects of wine consumption.
The experimental facilities
The development of Oenology teaching obliged the Faculty
to complete the facilities with vineyards and a cellar. The
municipality of Tarragona provided the Faculty of Oenology
with a 7 hectare plot . In this plot, a 5 hectare vineyard was
then developed. The vineyard was designed to fullfil both
teaching and research activities. On the one hand, 17 differ-
ent varieties were planted to allow the students to follow the
development of representative grape varieties. On the other
hand, several experimental plantations were undertaken to
analyze the plantation framework, water stress, and the ef-
fect of water supplementation on grape production and wine
quality. In addition, an ampelographic collection of 67 local
varieties are kept to test the oenological potential and pro-
jected future usage.
The experimental plots grow around a newly built 700 m2
winery (1995). The winery has a capacity for processing up
to 60 000 kg of grapes which normally yield about 45 000
litres of wine. This production is mostly used for teaching
purposes (wine made by the students as part of their curric-
ula) yet a reasonable amount is diverted into research. The
winery has a microvinification facility with hundred-litre vats
at a controlled temperature. These comprise an excellent re-
search complement to the industrial fermentations regularly
produced in the cellar.
Research laboratories in analytical chemistry, sensorial
analysis, molecular biology, microorganisms and plant bio-
chemistry, microbiology and food chemistry make up the fi-
nal set up of research resources linked to the Faculty of
Oenology. For sensorial analysis a tasting room for up to 40
INTEGRATED RESEARCH IN OENOLOGY*
* Albert Mas and Lluís Arola. Unitat d’Enologia del CeRTA.
Facultat d’Enologia, Universitat Rovira i Virgili. Ramón y Cajal, 70.
43005– Tarragona, Catalonia (Spain). Tel. 34 977 25 00 00. Fax: 34
977 25 03 47. Email: amb@ee.urv.es

people was built, with all the facilities regarding lighting and
isolation. A refrigerated wine storage room connecting to the
tasting room was also built.
The human factor: research groups
The research groups of the Department of Biochemistry and
Biotechnology are the most numerous within the Oenology
Unit. The research groups in Oenology from this department
are identified as Oenological Biochemistry and Oenological
Biotechnology.
The Oenological Biochemistry group deals with research
subjects in viticulture, fermentation and effects of wine com-
ponents on human health. In viticulture, applied research is
mostly carried out in the experimental plots and the water
availability to the grapevine is analyzed in terms of plant
growth and grape ripening. Soil structure and nutrient up-
take are strongly related to water availability and are studied
as related to leaf metabolism. Among the relevant factors for
wine quality in the ripening process, polyphenol synthesis is
a specific area of analysis [1]. Finally, cloning and sequenc-
ing Vitis genome and its expression completes the viticulture
research interests.
«Stuck» and «sluggish» fermentations are probably
among the problems of greatest import in modern oenology
[2]. A biochemical approach to this problem is taken by
members of the oenological biochemistry group in analysing
some of the most relevant factors, such as lipid metabolism
and the composition of the plasma membrane, membrane
carrier proteins, and nutrient uptake [3] on yeast exposed to
different stress factors such as ethanol, copper, organic
pesticides, etc [4, 5]. Due to the relevance of sparkling wine
in our immediate region, projects dealing with wine protein
characterisation related to foam formation and stability are
undertaken [6,7]. The influence of different winemaking
techniques on red colour extraction and stabilisation are
also analysed [8 – 11].
There is considerable evidence to suggest that small
amounts of alcoholic beverages reduce the risk of vascular
disease and total mortality in middle and old age. But it is not
clear whether wine is more protective than other alcoholic
beverages because it contains phenolic compounds. The
group has developed an animal experimental model which
can distinguish the effects of alcohol from the effects of the
non-alcoholic components present in wine [12], so as to de-
termine whether wine has effects other than those of alcohol
on cholesterol metabolism and oxidative status [13]. Due to
the complexity of in vivo studies; flavonoids, specially pro-
cyanidins, effects on cholesterol metabolism, fat reserves
[14] and oxidative stress are analyzed in cell lines. Also, the
distribution and metabolism of flavonoids are a subject of in-
tensive research in this group.
The oenological biotechnology group has focused their
research interest in the application of novel molecular biolo-
gy techniques to the different fields of oenology. Some of the
work was done in the field of viticulture by analising stress
factors in the growth and adaptation of grapevines [15, 16],
which led to the sequencing of some membrane proteins
[17]. Knowledge of these may be of industrial interest and
they are now protected by a registered patent [18]. Howev-
er, the main interest has been focused on industrial microor-
ganisms. The group has incorporated and developed mole-
cular biology techniques to identify different yeast species
[19], lactic acid [20] and acetic acid bacteria [21, 22]. These
techniques have been successfully applied to analyze yeast
population dynamics in different environmental conditions
during wine making. Thus, analysis of different winery envi-
ronments [23, 24], and oenological practices [25] have been
performed or are being peformed on yeast strain develop-
ment [26]. Also, the selection of yeast strains and their use
as fermentation starters has been carried out, already yield-
ing a comercial presentation [27].
Molecular analysis has also been applied to lactic acid
bacteria both for identification [20] and metabolic assess-
ment as response to different environmental situations. The
influence of compounds present in wine on kinetics of malo-
lactic fermentation and physiology of malolactic bacteria
have been studied, such as copper and other pesticides
[28, 29], fatty acids, sulphur dioxide, and phenolic com-
pounds [30]. The presence of bacteriophages in Oenococ-
cus oeni and its relation to lysogeny and stuck malolactic
fermentation has also been studied [31]. Additionally, sever-
al strains of malolactic bacteria have been isolated for use
as starters [32].
The Analytical Chemistry of Wine and Aliments Research
group which is a part of the Department of Analytical and Or-
ganic Chemistry, has been very active in developing analyti-
cal techniques for aroma and pesticide analysis in wine us-
ing innovative and conventional methods [4, 33-40]. Method
validation for wine and must analysis has been carried out
as well as assessment of the implementation of quality sys-
tems in different food analysis laboratories.
The Food Technology research group of the Department of
Chemical Engineering has developed research activities in
membrane separation techniques in wine and beer produc-
tion, characterisation of membrane fouling during microfiltra-
tion and new processes to prevent and control this [41-43].
Also, the use of continuous processes for stabilisation and
decoloration as well as the effects of must clarification on al-
coholic fermentation has been the focus of its research [44].
However, none of these actions will be effective if the pro-
fessional sector is not involved. Several research projects
are underway, although the researchers and the research
groups do not have the time to devote to administration and
the amount of faceto-face contact development required.
Thus, an oenologist acts as a conduit of knowledge, in order
to best utilize the Faculty’s human resources and match
them to the needs of the wine sector while promoting and
maintaining contact within the sector.
All the Faculty’s researchers are very active in a wide vari-
ety of knowledge transfer activities, such as conferences,
seminars and meetings. A yearly symposium is held in Vi-
lafranca del Penedès which presents the most relevant ad-
366 Albert Mas and Lluís Arola

vances in oenological research done by the group re-
searchers. The conferences are collected in a publication
series, Oenology Today, published by the Oenology Unit of
the Reference Centre for Food Technology
References
[1] Nadal, M., Arola, Ll. Effects of limited irrigation on the
composition of must and wine of Cabernet Sauvignon
under semi-arid conditions. Vitis 34 (3), 151-154, 1995.
[2] Zamora, F. Los problemas de fermentación. In «Temas
actuales en la microbiología enológica», Ed. Ayunta-
miento de Haro, pp 51-79, 1994. ISBN: 84-7359-467-3.
[3] Zamora, F., Fort, F., Fuguet, J., Bordons, A., Arola, Ll.
Influence de certains facteurs lors de la fermentation
alcoolique sur la prise de glucose par les levures. In
«Œnologie 95», Ed. A. Lonvaud-Funel, Lavoisiser,
Paris, 1995, pp 167-171. ISBN: 2-7430-0083-X.
[4] Sala, C., Fort, F., Busto, O., Zamora, F., Arola, Ll.,
Guasch, J. Fate of some common pesticides during
vinification process J. Agr. Food Chem. 44, 3668-
3671,1996
[5] Fort, F., Sala, C., Busto, O., Arola, Ll., Bordons, A.,
Guasch, J., Zamora, F. Presenza di pesticidi in vinifi-
cazione ed inibizione della fermentazione alcolica. Vi-
gnevini, 7/8, 42-44, 1999.
[6] Canals, J.M., Arola, Ll., Zamora, F. Protein fraction
analysis of white wine by FPLC. Am. J. Enol. Vitic., 49,
383-388, 1998.
[7] Zamora, F., Luengo, G., Margalef, P., Magriña, M., Aro-
la, Ll. Efecto del sangrado sobre el color y la composi-
ción en compuestos fenólicos del vino tinto. Rev. Esp.
Cien. Tecnol. Alimen., 34, 663-671, 1994
[8] Vivas, N., Zamora, F., Glories, Y. Etude des
phénomenes d’oxydorreduction dans les vins. Mise au
point d’une méthode rapide de mesure du potentiel
d’oxydorreduction J. Int. Sci. Vigne Vin, 26, 271-285,
1992
[9] Vivas, N., Zamora, F., Glories, Y. Incidence de certains
facteurs sur la consommation de l’oxygène et sur le po-
tentiel d’oxydorreduction dans les vins. J. Int. Sci. Vi-
gne Vin, 27, 23-34, 1993
[10] Vivas, N., Glories, Y., Bertrand, A., Zamora, F. Principe
et méthode de mesure du potentiel d’oxydoréduction
dans les vins. Bull. O.I.V., 785-786, 618-633, 1996
[11] Vivas, N., Saint-Cricq de Gaujeac, N., Zamora, F. Ex-
perimental principe to evaluate the degree of oxyda-
tion or reduction balance in wines. J. Sci. Tech. Tonnel-
lerie, 5, 65-76, 1999.
[12] Arola Ll., Roig R., Cascón E., Brunet M.J., Fornós N.,
Sabaté M., Raga X., Batista J., Salvadó M.J., Bladé C.
Model for voluntary wine and alcohol consumption in
rats. Physiol. Behav. 1997, 62: 353-357.
Integrated research in oenology 367
Department Research Permanent Graduate
Group Members Students
Biochemistry and Biotechnology Oenological Biochemistry Lluís Arola Montse Pinent
Fernando Zamora Noemí Ferrer
Joan Miquel Canals Guillem Vanrell
Montserrat Nadal Pedro Cabanillas
Francesca Fort Miriam Lampreave
Cinta Bladé Josep Valls
Josepa Salvadó Bernardino García
Anna Ardèvol Francesc Puiggrós
Isabel Baiges
Oenological Biotechnology Albert Mas Maria Jesús Torija
José M. Guillamón Angel González
Nicolas Rozes Gemma Beltran
Albert Bordons Maite Novo
Magda Constantí Ramon Carreté
Montserrat Poblet Gerrie Garcia
Cristina Reguant
Analytical and Organic Chemistry Wine and Food Josep Guasch Montse Mestres
Analytical Chemistry Olga Busto M. Pilar Martí
Cristina Sala
M. Dolors Jornet
Eva Brull
Marc Rius
Nuria Martorell
Noemí Carrasco
Chemical Enginneering Food Technology Montserrat Ferrando Vesselina Pachova
Carme Güell Justyna Warczok
Francisco López
Table 1. The research groups in the Oenology Unit of the Centre for Reference for Food Technology

[13] Roig R., Cascón E., Arola Ll., Bladé C., Salvadó M.J.
Moderate red wine consumption protects the rat
against oxidation in vivo. Life Sci. 1999, 64: 1517-1524.
[14] Ardévol, A., Bladé, C., Salvadó, M.J, Arola Ll. Changes
in lipolysis and hormone-sensitive lipase expression
caused by procyanidins in 3T3-L1 adipocytes. Int J.
Obesity, 24: 319-324, 2000.
[15] Romeu, A., Mas, A. Effects of copper exposure in tis-
sue cultured Vitis vinifera . J. Agr. Food Chem, 47,
2519-2522, 1999
[16] Llorens, N., Arola, Ll., Bladé C., Mas A. Effects of cop-
per exposure upon nitrogen metabolism in tissue cul-
tured Vitis vinifera. Plant Sci. in press.
[17] Baiges, I., Schaeffner, A.R., Mas, A. Sequences of Vitis
berlandieri x Vitis rupestris putative aquaporins: PIP 1-
1 (Accesion number AF141643), PIP1-2 (Accesion
number AF141898), PIP 1-3 (Accesion number
AF141899), PIP 2-1 (Accesion number AF141642), PIP
2-2 (Accesion number AF141900), TIP 1 (Accesion
number AF271661), TIP 2 (Accesion number
AF271662), TIP 3(Accesion number AF271660).
[18] Baiges, I., Schaeffner, A.R., Mas, A. Nukleinsäuren
und hieraus abgeleitete Oligonukleotide zur spezifis-
chen Amplifikation und zum spezifischen Nachweis
von Aquaporin-Genen aus Vitis vinifera. Patent register
number 10011480.6 (Germany). 2000
[19] Guillamón, J.M., Barrio, E., Huerta, T., Querol, A. Rapid
characterization of four species of the Saccharomyces
sensu stricto complex according to mitochondrial DNA
patterns, Int. J. Syst. Bacteriol., 44, 708-714, 1994
[20] Zapparoli, G, Reguant, C., Bordons, A., Torriani, S.,
Dellaglio, F. Genomic DNA fingerprinting of Oenococ-
cus oeni strains by pulsed-field gel electrophoresis
and randomly PCR amplified polymorphic DNA, Curr.
Microbiol., in press.
[21] Poblet, M., Rozés, N., Guillamón, J.M., Mas, A. Identifi-
cation of acetic acid bacteria bacteria by restriction
fragment length polymorphism analysis of a PCR-am-
plified fragment of the gene coding for 16S rRNA. Lett.
Appl. Microbiol., 30, 1-7, 2000
[22] Ruiz, A. Poblet, M. Mas, A. Guillamón JM. Identification
of acetic acid bacteria by RFLP of the PCR-amplified
16S rDNA and 16S-23S rDNA intergenic spacer. Int. J.
Syst. Evol. Microbiol., in press
[23] Constantí, M., Poblet Icart, M., Arola, Ll., Mas, A., Guil-
lamón, J.M. Analysis of yeast populations during alco-
holic fermentation of wine in a newly established win-
ery. Am. J. Enol. Vitic. , 48, 339-344, 1997
[24] Torija, M.J., Ruiz, A., Martí, M., Beltran, G., Llauradó, J.,
Poblet, M., Rozès, N., Guillamón, J.M., Mas, A. Étude
de l’évolution des levures et bactéries acétiques par
l’utilisation de techniques de biologie moléculaire lors
de fermentations spontanées et inoculées . Oenologie
99 , A. Lonvaud ed, Tech&Doc, 2000 , pp 399-402.
[25] Constantí, M., Reguant, C., Poblet, M., Zamora, F.,
Mas, A., Guillamón, J.M. Molecular analysis of yeast
population dynamis: Effect of sulphur dioxide and the
inoculum in must fermentation. Int. J. Food Microbiol. ,
41, 169-175, 1998
[26] Llauradó, J., Constantí, M., Rozés, N., Mas, A.,
Velázquez, R., Bobet, R. Fermentaciones a bajas tem-
peraturas (13C): Efectos de las cepas de levaduras y
la adición de nutrientes. Alimentación. Equipos y Tec-
nología, XIX, 87-92, 2000
[27] Torija, M.J., Rozès, N., Guillamón, J.M., Mas, A. Sac-
charomyces cerevisiae. Patent register CECT 11462.
Date: 1/11/99. Comercial presentation RV1 (Lallemand
Inc).
[28] Bordons, A., Masqué, M.C., Vidal, M.T. Isolation and
selection of malolactic bacteria and effect of pesti-
cides. The management of malolactic fermentation
and quality of wine. Les entretiens scientifiques Lalle-
mand, 51-56, Verona, 1998
[29] Vidal, M.T., Constantí, M., Bordons, A. Effeto del rame
e dei pesticidi sulla fermentaziones malolattica.Vi-
gnevini , 7/8, 50-53, 1999
[30] Reguant, C., Bordons, A., Arola, L., Rozès, R. Influence
of phenolic compounds on the physiology of Oenococ-
cus oeni from wine. J. Appl. Microbiol., 88, 1065-1071,
2000
[31] Poblet, M., Bordons, A., Lonvaud-Funel, A. Lysogeny
of Oenococcus oeni and study of their induced bacte-
riophages. Curr. Microbiol., 36, 365-369, 1998
[32] Masqué, M.C., Bordons, A. Isolation and selection of
malolactic bacteria from southern Catalan wines. J.
Wine Res., 7, 91-101, 1996.
[33] Busto, O., Guasch, J., Borrull, F. Biogenic amines in
wine: A review of analytical methods. J. Int. Sci. Vigne
Vin, 30, 85-101, 1996.
[34] Busto, O., Guasch, J., Borrull, F. Determination of bio-
genic amines in wine after pre-column derivatisation
with 6-aminoquinolyl-n-hydroxysuccinimidyl-carba-
mate. J. Chromatogr., 737, 205-213, 1996.
[35] Busto, O., Miracle, M., Guasch, J., Borrull, F. Solid
phase extraction of biogenic amines from wine before
chromatographic analysis of their AQC-derivatives. J.
Liq. Chromatogr., 20, 743-755, 1997
[36] Sala, C., Busto, O., Guasch, J. A quick capillary gas
chromatographic method for determining usual pesti-
cides in must and wines. Chromatographia, 44, 320-
324, 1997
[37] Mestres, M., Busto, O., Guasch, J. Chromatographic
analysis of volatile sulphur compounds in wines using
the static headspace technique with flame photometric
detection. J. Chromatogr., 773, 261-269, 1997
[38] Mestres, M., Busto, O., Guasch, J. Headspace solid
phase microextraction analysis of volatile sulphides
and disulphides in wine aroma. J. Chromatogr., 808,
211-218, 1998
[39] Mestres, M., Sala, C., Martí, MP., Busto, O., Guasch, J.
Headspace solid phase microextraction of volatile sul-
phides and disulphides using carboxenpolydimethyl-
siloxane fibers in the analysis of wine aroma. J. Chro-
matogr., 835, 137-144, 1999
368 Albert Mas and Lluís Arola

[40] Mestres, M., Martí, MP., Busto, O., Guasch, J. Simulta-
neous analysis of thiols, sulphides and disulphides in
wine by headspace solid phase microextraction gas
chromatography. J. Chromatogr., 808, 211-218, 1999.
[41] Güell, C., Davis, R.H. Membrane fouling during micro-
filtration of protein mixtures. J. Membrane Sci., 119,
269, 1996.
[42] Czejak, P., López, F., Güell, C. Characterization of
membrane fouling caused by protein fraction in wine.
Am. J. Enol. Vitic., 48, 388, 1997.
[43] Güell, C., Czekaj, P,. Davis, R.H. Microfiltration of pro-
tein mixtures and the effect of yeast on membrane foul-
ing. J. Membrane Sci., 155, 113-122, 1999.
[44] Ferrando, M., Güell, C., López, F. Industrial wine mak-
ing: Comparison of must clarification treatments. J.
Agr. Food Chem., 46, 1523-1528, 1998
Integrated research in oenology 369
About the authors
Lluís Arola obtained his PhD. in Biochemistry at the University of Barcelona in 1978. His research has primarily dealt with me-
tabolism and nutrition. In 1988, he was the first director of the Oenology School. Since that time he has been actively involved in
biochemistry of fermentations and the effects of some wine components in lipid and oxidative metabolism.
Albert Mas is the present Dean of the Faculty of Oenology. He obtained the PhD. in Biochemistry from the University of
Barcelona in 1984. In previous research he worked with metal metabolism, but since his incorporation into the Oenology School
(1992) his investigations have focused on the effects of copper on grapevine metabolism and on development of molecular bi-
ology techniques for analysing microorganism population dynamics during fermentation.