1. Grape berry yeast communities: Influence of fungicide
treatments
Vesna Milanović, Francesca Comitini, Maurizio Ciani.
International Journal of Food Microbiology
Estefania Jaramillo Salazar
Luis Camilo Maldonado
3rd Semester
Medicine students
U.P.Ba

2. INTRODUCTION:
• The presence of yeast and bacterial species on grape berry surfaces dependent
on several environmental factors, such as: application of agrochemical
treatments… consequently the good fungicide selection is essential and the
effects of new generation fungicides are not well known.

3. INTRODUCTION:
• there is great concern about the impact of fungicides on human health,
therefore there is an increasing trend towards consumption of organic food
‘Natural’ , They carried out qualitative and quantitative evaluation.
• of the yeast communities under treatment based on formulations containing
copper and sulphur compounds , and under treatment with conventional
fungicides, using both culture-dependent and -independent methods.

4. YEAST GENERALITIES:
• What is? Yeasts are microscopic fungi, unicellular.
• Where is? Is found naturally on the surface of plants.
• what shape they have?
• What is your cell structure?
• How they reproduce?
Yeasts reproduce asexually by budding
• and sexually by ascospores

5. YEAST GENERALITIES:
• What is their utility ? They Ferment various organic bodies, especially
sugars or carbohydrates, producing multiple substances.
• Also to express recombinant proteins because its easy to use
• How do they act? Produce enzymes capable
• of decomposing a variety of substrates, primarily sugars.
•
An example of yeast: Saccharomyces cerevisiae

6. FUNGICIDES GENERALITIES:
• What are they?
Fungicides are pesticides that act on pathogenic fungi.
What types of fungicides are according to their action?
are two types of fungicides:
1 -Contact
2 -systemic
• What types of fungicides are according to their field of application?
• Use in seed coatings.
• Use for soil disinfection.
• For application on plants.

7. FUNGICIDES GENERALITIES:
• how do they act?
Most fungicides are sprayed or dusted farm on seeds, leaves or fruit.
•
What types of fungicides are according to their composition?
organic fungicides and Conventional fungicides.

8. GRAPE BERRY GENERALITIES:
• What is ? the grape is a fruit
• how they grow? They Grow in clusters of between 6 and 300 grapes.
•
What color can have? They can be black, purple, yellow, gold, purple, pink,
brown, orange or white.
•
What are the main grape producing countries?: Australia, Europe, USA and
South Africa.

9. GRAPE BERRY GENERALITIES:
• What is the grape content? The grape conteins 80% water, fiber,
• many calories, carbohydrates (glucose and fructose = sugar), Vitamins,
minerals…
• What are grape properties?
- Debugger and detoxifying.
- help to Intestinal transit problems.
- Immune system: produce red and white blood cells, helps to antibody
formation.
-Brain function.
-Cardiovascular and degenerative problems….

10. LINKAGE:
• the quality of grape berry is strongly influenced by the kind of
treatment used, in the yeast communities on grape surfaces
can produce either beneficial or harmful influence.

11. GENERAL OBJECTIVE:
• the goal of this work is the comparison between organic and
conventional treatment in the vineyards.

12. MATERIALES Y MÉTODOS
• El medio de cultivo:
es una Solución que cuenta con los nutrientes necesarios para,
multiplicar, aislar e identificar los microorganismos (bajo
condiciones favorables de temperatura y pH)
fue usado para realizar un adecuado recuento de las células
aisladas de las levaduras

13. MATERIALES Y MÉTODOS
• Muestreo
• 6 racimos (3kg c/u) uvas de un viñedo óganico
tratadas con mezcla de burdeos
• 6 racimos (3kg c/u) Uvas de un viñedo convencional
tratadas con productos químicos
• Luego se separaron en bolsas donde se homogenizó cada contenido
manualmente y se transfirieron a matraces de 0,5L proceso de
fermentación .
• las muestras fueron tomadas al principio, en la mitad y al final de la
fermentación

14. MATERIALES Y MÉTODOS
• Muestreo.
• Las muestras fueron llevadas al medio de cultivo 5 días
fueron seleccionadas :
• Según sus aspectos morfológicos 30 y 40 colonias c/muestra
• Según la proporción de las levaduras 50 y 300 colonias c/muestra

15. MATERIALES Y MÉTODOS
• Extracción del DNA
Son las diferentes técnicas que empleo para extraer el DNA y de
ésta manera poder analizar el genoma
Se utilizó para identificar regiones en el DNA de las levaduras.

16. MATERIALES Y MÉTODOS
• PCR:
• Es una técnica de amplificación enzimática de secuencias específicas de
DNA.
• Se utilizó para aumentar el segmento específico de ADN extraído de la S.
cerevisiae

17. RESULTADOS:
Fig.1 Total yeast counts of all of the samples from the organic and conventional vineyards
at the beginning of the fermentation. Inset: Total yeast counts shown collectively according
to organic (grey) and conventional (black) vineyards. .

18. RESULTADOS:
•Un total de 1.057 colonias de levadura fueron aislados de estos doce
muestras que se recogieron durante las tres fases de análisis: medias, al
principio y al final de la fermentación.
•Las Muestras obtenidas por DGGE fueron sometidas a UPGMA ; para
evaluar las diferencias entre la viña tratada con fungicida convencional y
fungicida orgánico.

19. RESULTADOS:
Un total de
1.057 colonias
de levadura
fueron aislados
de estos doce
muestras que
se recogieron
durante las tres
fases de
análisis: medias
principio y al
final de la
fermentación

20. RESULTADOS:

21. RESULTADOS:
Como se indicó anteriormente,
utilizando el cultivo dependiente
de análisis en el inicio de la
fermentación, se produjo una
menor diversidad de levaduras en
las muestras de la viña orgánica
que desde el viñedo convencional,
y esto se confirmó por análisis de
DGGE.
poblaciones inferiores a 10 3 UFC /
ml; no se distingue en la electroforesis

22. RESULTADOS:
La figura. 3. análisis
UPGMA cluster de los
perfiles de DGGE de las
muestras recogidas de
los orgánicos (1O-6O) y
convencional (1C-6C)
viñedos en el inicio de la
fermentación, basado en
la banda de
revelado. Escala: las
distancias promedio
entre las muestras. BF =
inicio de la fermentación.

23. DISCUSSION
SOURCE AFFIRMATION AGREE/DISAGREE
Barba The frequency and relative high Agree
doses of these organic products
might have antimicrobial effects on
the yeast on the grape berries. A
previous study indicated that
fungicides based on sulphur
compounds has no negative
effects on yeast, while those
formulated on copper compounds
can significantly inhibit growth of S.
cerevisiae.
Crowdy Moreover, the higher yeast density Agree
on the grape berry surfaces with
the conventional fungicides might
be explained by the fact that
together with the active
ingredients,these chemicals can
contain additives, such as
surfactants,emulsifiers,dispersants,
solubilisers, solvents, diluents,
preservatives and thickeners,
which are used to enhance the
efficacy and stability of
the fungicides.

24. DISCUSSION
SOURCE AFFIRMATION AGREE/DISAGREE
Oliva These data are related to Agree
the previously reported
yeast abundance,and
they can be justified by an
absence or a reduced
selective effect on the
yeast flora of the new
generation fungicides
used.(for example:
famoxadona, fenhexmid,
quinoxifeno)
Martini As some studies have Disagree
also not been able to
isolate this species from
healthy, mature grapes,
these observations have
raised speculation and
controversy as to the
origin of S. cerevisiae in
wine production

25. CONCLUTIONS
The fungicides can be enhance the
The fungicides can be enhance the
production and alcoholic
production and alcoholic
fermentation,but could be decrease
fermentation,but could be decrease
specific yeast that are important to
specific yeast that are important to
fermentation of that wine
fermentation of that wine
for healtyness Its so important have aa
for healtyness Its so important have
good and extrictly control with
good and extrictly control with
fungicides of the grapes berry surface
fungicides of the grapes berry surface
because we can sickness if it was
because we can sickness if it was
damage or toxic
damage or toxic

26. CONCLUTIONS
• Organic fungicides are more effect
to decrease yeast than
convencional
Sometimes the production of
Sometimes the production of
specific wine could be affected by
specific wine could be affected by
fungicide
fungicide

27. MAPA CONCEPTUAL ESTEFANIA:

28. MAPA CONCEPTUAL CAMILO:

29. GRACIAS