Professional Documents
Culture Documents
Actualizacin de conocimientos
Christophe Gerland
christophe.gerland@intellioeno.com
www.intellioeno.wix.com/intellioeno
SUMMARIO DE LA PRESENTACION
Los microorganismos mas peligrosos y los puntos llave en sus
manejos
Lo que piden los clientes y la legislacion
Importancia de los biofilms
Los puntos llave al momento del embotellamento
Parte 1
Los microorganismos mas peligrosos
Los puntos llave en sus manejos
Enemigos vinicos n1
Brettanomyces
Lactobacillus
Zygosaccharomyces bailii
Enemigos vinicos n1
Brettanomyces
Lactobacillus
+ bacterias aceticas
+ Pediococcus
Levaduras de refermentacion
Se detectan Brettanomyces en la
lina de embotellamento
ATP
BRETTANOMYCES
valvula filtro
ausencia
bec rinceuse
10
Ausencia
bec rinceuse
Ausencia
BEC 1
28
presencia *
BEC 12
35
presencia *
BEC 19
59
presencia *
BEC 29
84
non effectu
BOUCHEUSE
15
ausencia
BRETTANOMYCES : AUMENTO DE SU
RESISTENCIA AL SULFUROSO
Ms exactamente : identificacin cada vez ms frecuente de cepas muy
resistentes
Curtin , Australia
Aislamiento de 244 cepas de Brett en Australia
3 genotipos ms frecuentes
Resistencia al sulfuroso respectivamente de 30, 40 y 75 mg/L (valor mximo
donde se observa crecimiento en medio sinttico pH no indicado)
Edwards, USA
Identificacin de 2 cepas de Brett. que pueden multiplicarse en un vino de
Syrah despus de adicionar 0,89 mg/L de SO2 molecular activo
Despus de 1 da, no se detectan en medio de cultivo (unicamente en
epifluorescencia = viable pero no cultivable) , despus de 10 dias multiplicacin
para volver al nivel inicial de 1 milln/mL
Coulon e Lonvaud, Bordeaux (Symposium international oenologie Bordeaux
2011)
Han islado en terroirs de Bordeaux diversas cepas de Brett resistentes a mas
de 0.8 mg/L de SO2 molecular activo
coutarico
Aspergillus niger
cinnamyl esterasa
acido cafeico
Levaduras POF+
Cinnamato
decarboxilasa
4-vinil-catecol
fertarico
Levaduras ?
cinnamyl esterasa
acide coumarique
acide frulique
Brettanomyces
Cinnamato dcarboxilasa
4-vinil-fenol
4-vinil-guaacol
Brettanomyces
vinil fenol reductasa
4-etil-catecol
4-etil-phnol
4-etil-guaacol
2013
Osborne, 2013
CAPACIDAD DIFERENTE DE LAS CEPAS DE OENOCOCCUS OENI
A TRANSFORMAR COUTARICO EN COUMARICO
Edwards, 2013
LAS CEPAS DE BRETTANOMYCES NO PUEDEN HACER ESTA TRANSFORMACION
Caftarico
coutarico
Aspergillus niger
cinnamyl estrase
acido cafeico
Levures POF+
Cinnamate
dcarboxylase
Edwards, 2013
fertarico
Levures ?
cinnamyl estrase
acide coumarique
Brettanomyces
cinnamyl estrase
acide frulique
Brettanomyces
Cinnamate dcarboxylase
4-vinil-fenol
4-vinil-guaacol
Brettanomyces
vinyl phnol reductase
Interes de hacer la
Analisis
Del coumarico libre
4-etil-catecol
4-etil-phnol
4-etil-guaacol
Vapor
Analisis de antes/despues del
proceso
1,00E+06
1,00E+06
1,00E+05
1,00E+05
Avant
1,00E+04
Aprs
1,00E+03
Finition
1,00E+02
Brett en UFC/mL
Ptri (VC)
Cytomtrie (VNC)
1,00E+04
1,00E+03
1,00E+02
1,00E+01
1,00E+01
1,00E+00
J+7
1,00E+00
Barrique 1
Barrique 2 Barrique 3
(Mchage) (Oxygne -)
J+28
Barrique 1
J+7
J+28
Barrique 2
(Mchage)
J+7
J+28
Barrique 3
(Oxygne - )
5 min
leau
ozone
850-900
ppm
Agua ozonizada
Analisis de la madera
antes/despues del tratamiento
Ptri (VC)
Cytomtrie (VNC)
1,00E+06
Avant
1,00E+04
Aprs
1,00E+03
Finition
Brett en UFC/mL
1,00E+05
1,00E+05
1,00E+04
1,00E+03
1,00E+02
1,00E+02
1,00E+01
1,00E+01
1,00E+00
J+7
1,00E+00
J+28
Barrique 1
J+7
J+28
Barrique 2
(Mchage)
J+7
J+28
Barrique 3
(Oxygne -)
Vapor
Mechado
Ultra-sonidos
Agua caliente
Unicamente en
superficia
Entran en profundidad
Sosa/
permanganato
Agua ozonizada
No desinfectan totalmente
La superficia
Wood seeker se
conecta con una
perceuse y permite
de tomar muestra
de roble hasta 1 cm
de profundidad
Inoculacion de
sniffbrett y
incubacion. Lectura
de 1 a 10 dias,
acopleado con
citometria a partir
de 3 dias.
2
El resultado permite de
ver y despues optimizar
el protocolo de limpiezadesinfecion de las
barricas.
Alta pression
UVB
Micro-ondas
Ultra-sonidos
A CONSIDERAR
ANTES DEL
EMBOTELLADO
Identificar los vinos con riesgo : por la analisis fina del fructosa
FRUCTOSE > 0,100 g/L
Ni 4EP ni Brett
Mais vin risque si arrive une contamination
Heterofermentativas : hilgardii,
brevis,
Picadura lctica
Enfermedad del rebote o tourne
Enfermedad del amargor (glicerol acroleina)
Gusto a ratn (produccin de Tetrahidroxipiridina)
Gusto a raton
Acetobacter
Quinto microorganismo
Pediococcus
Brettanomyces
Zygosaccharomyces y Saccharomycodes
Lactobacillus
Acetobacter
Pediococcus
Alta pression
UVB
Micro-ondas
Ultra-sonidos
Agua ozonizada o ozono gaz ?
que tipo ?
3,50E+03
3,00E+03
Brett (/mL)
2,50E+03
Non filtr
2,00E+03
10 m
1,50E+03
4 m
1,00E+03
1 m
5,00E+02
0,3 m
0,00E+00
1993
1994
Renouf, Lonvaud
Am. J. Enol. Vitic.
58:3 (2007)
1995
2000
1500
Non filtr
10 m
4 m
1 m
500
0,3 m
0
1993
1994
1995
6,00E+03
5,00E+03
4,00E+03
3,60E+03
no filtrado
Brett(/mL) 3,00E+03
7 m
2,00E+03
1,40E+03
1,00E+03
4 m
1,25E+02
Renouf, Lonvaud
Am. J. Enol. Vitic.
58:3 (2007)
0,00E+00
1996
1997
1998
4000
3500
3000
2500
4-EP (g/L) 2000
1500
1000
Non filtr
7 m
4 m
500
0
1996
1997
1998
Evaluation of the inhibitory effect of dimethyl dicarbonate (DMDC) against wine microorganisms
A. Costa,
A. Barata,
M. Malfeito-Ferreira<img
V. Loureiro
Several microbial species associated with wine were challenged against increasing
concentrations of dimethyl dicarbonate (DMDC). The concentration inducing complete
cell death upon addition to red wine was regarded as the minimum inhibitory
concentration (MIC). In dry red wines with 12% (v/v) ethanol and pH 3.50, the
inactivation depended on the initial cell concentration. For an initial inoculum of
500 CFU/ml, the MIC of the yeasts species Schizosaccharomyces pombe, Dekkera
bruxellensis, Saccharomyces cerevisiae and Pichia guilliermondii was 100 mg/l. The
most sensitive strains belong to Zygosaccharomyces bailii, Zygoascus hellenicus and
Lachancea thermotolerans, with MIC of 25 mg/l DMDC. For inoculation rates of about
106 CFU/ml, the maximum dose of DMDC legally authorised (200 mg/l) was not
effective against the most resistant species. The addition of 100 mg/l potassium
metabisulphite (PMB), equivalent to 1 mg/l molecular sulphur dioxide, increased the
inactivation effect of 100 mg/l DMDC over initial yeast populations of 106 CFU/ml but
did not fully kill S. pombe and S. cerevisiae.
Lactic acid and acetic acid bacteria were not killed by the addition of 300 mg/l of
DMDC. Trials performed in wines before bottling showed that in most samples
indigenous bacterial populations were not affected by 200 mg/l DMDC. Therefore,
under winery practice, DMDC at the maximum dose legally permitted may be regarded
as an efficient preservative to control low contamination rates of yeasts but ineffective
against lactic acid and acetic acid bacteria.
chitosan
Alta pression
tecnicas fisicas (Bartowsky, 2008)
UVB (Tebaldi, 2011)
Campos pulsados (Poulard, 2011)
Micro-ondas
Ultra-sonidos (Jiranek et al. 2008)
Embotellado al caliente
3.5.4. EMBOUTEILLAGE A CHAUD (OENO 9/97)
Dfinition :
Mise en bouteille du vin pralablement rchauff et bouchage
immdiat de la bouteille.
Objectifs :
a) La stabilisation biologique du vin ;
b) L'limination de l'oxygne ;
c) La stabilisation physico-chimique.
Prescription:
Le vin ne doit pas tre rchauff plus de 45C.
Recommandation de l'OIV :
Admis.
ALTA PRESSION
Alta pression
UVB
Micro-ondas
Ultra-sonidos
TURBIDITY (NTU)
50
40
WINE TREATED WITH LYSOZYME
30
20
10
0
0
10
12
ver www.lysoclub.com
por el protocolo completo
make a well
analisis del
14/8/2012
REF.
DO 450
lisozima (mg/L)
VINO
B5
0.358
0,004
B6
0.355
0,004
R6
1.914
0,307
CRIANZA MALBEC
R7
0.521
0,036
CRIANZA TINTO
R8
2.572
>0,4
R9
0.528
0,037
R10
1.234
0,175
R11
2.645
>0,4
CRIANZA B1204
R12
0.488
0,03
R13
0.358
0,004
R14
2.209
0,364
0,163
R15
2.156
0,354
0,081
En verde los vinos que son debajo de la limita oiv de 0,25 mg/L
Parte 2
Lo que piden los clientes
Obligacion de limpieza
Y desinfecion antes
De poner un vino en
Un deposito, botella,..
Parte 3
Como detectar y eliminar los biofilms
Parte 4
Los puntos llave al momento del embotellamento
Vinos
Materiales
Aire
Adaptacion del processo
The airborne yeast flora associated with four different areas in a Spanish
winery was analyzed monthly over the course of one year (February 2008
January 2009), in order to characterize the yeasts in this particular ecosystem,
an aspect relatively unknown until now. The sampling areas were: vinification,
bottling, cask aging and bottle cellar aging. 382 yeasts were identified from
two different culture media: one generic for yeasts and the other selective for
the Brettanomyces/Dekkera genus. The levels of yeasts present in the air
throughout the year were low and the highest presence of yeasts was in the
bottling area. Most of the yeasts isolated were members of the nonSaccharomyces groups. Yeasts of the Saccharomyces cerevisiae species were
only detected in the air in the vinification area during the vinification period.
Sporidiobolus and Cryptococcus were the dominant genera in the air and they
were found continuously in every area analyzed. The spoilage yeast
Brettanomyces/Dekkera was not detected in any sampling made.
PRECAUCION MINIMA : poner pression positiva en la sala de embotellamento
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