Stephanie Yin and Karen Lin

January 25, 2011

Science Fair
Introduction to Science Fair
We first became aware of the natural cleaning effects of wine through the online case

study, “Wine Kills Several Types of Foodborne Pathogens”. The article discusses how the

acidity of wines may contribute to its antimicrobial effect on foodborne pathogens such as E.

coli (Oregon State U, 2002). Researchers simulated a model of the stomach and tested wine,

specifically white wine, because it is more acidic than red wine, and discovered that it reduced

the levels of E. coli in the stomach. Consequently, we revised our initial procedure to test the

effect of various types of non-alcoholic wines on E. coli grown in LB agar plates. Escherichia

coli (E. coli) are members of the Enterobacteriaceae family of gamma-proteobacteria that

inhabit the lower intestines, known as the gut flora. The presence of this rod-shaped bacteria,

along with other species of bacteria in our intestinal tract, provide us with many necessary

Vitamin K and B-complex vitamins. However, enterohemorrhagic strains of E. coli may cause

severe anemia or kidney failure, which can lead to death. Other rare strains can cause urinary

tract infections. E. coli can be passed from person to person, but serious E. coli infections are

more often linked to food containing the bacteria, such as undercooked meat, contaminated

vegetables, and unpasteurized fruit juice. Although heat is known to kill E. coli, Microbiologists

Mark Daeschel and Jessica Just at Oregon State University (OSU) discovered that wine,

especially white wine, inactivated virulent bugs (called pathogens) like E. coli. After managing

to isolate the properties in wine that killed the bugs, they found that it is the malic and tartaric

acid that kill bacteria, rather than anything present in the alcohol (2002).
 Wine is a natural antimicrobial, and scientists at OSU are developing a new disinfectant

containing wine to fight off microbes like bacteria. The major grape acids in wine are tartaric and

malic acid, with citric being among the minor ones. Tartaric acid is found in almost no fruit but

the grape and here it is the predominate acid. It is important because it is the strongest and most

voluminous acid present in grape wines and, with its potassium and calcium salts, largely

controls the effective acidity (pH) of such wines. Malic acid is one of the most widespread acids

among the many fruits and vegetables from which wines are made. Although acid is not required

for yeast to reproduce and convert sugar into alcohol and CO2, it is desired in terms of flavor.

More importantly, the role of acids in winemaking comes from their ability to stop, or at least

retard, the growth of many potentially harmful microorganisms that would spoil the wine itself.

"Antimicrobial" is a word that describes both natural and man-made substances,

including antibiotics and disinfectants, that can kill or slow down the growth of bacteria and

viruses and other microorganisms. However, sometimes microorganisms can develop a

resistance over time to an antimicrobial substance, so that it no longer stops microbes from

growing. Naturally occurring alternatives could help address this problem. We've all heard

advice from seasoned travelers that it's often better to drink the wine than the water in a foreign

country. There is some truth to the adage. Wine drinkers, for example, rarely suffer from food

poisoning, such as salmonella, the leading cause of food poisoning. The wine's natural acids

combine with its alcohol content to kill bacteria. The OSU scientists discovered it also kills

bacteria on a kitchen counter, for example. As an added benefit, using wine as an anti-microbial,

or as a natural food preservative, cuts down on the amount of chemicals in the environment, and

on the long and costly development process for chemically based food preservatives.
 If we use substances containing proanthocyanidins, malic acid, and tartaric acid, such as

non-alcoholic wines, then the concentration of E. coli in LB agar plates will decrease.

Proanthocyanidins form a group of chemical compounds that are found in many plates which are

an important part of the human diet. These compounds are termed flavanoids which is a category

of plant chemicals. Proanthocyanidins are found in teas, black currant, bilberry, cranberry, grape

seed, and grape skin. There has been a lot of speculation about proanthocyanidins having a

positive effect on capillary fragility, retinopathy, sunburn, varicose veins, and pancreatic

insufficiency, but there has not been any clinical evidence to support these claims.

We plan to test the effect of non-alcoholic wine on E. coli by using a method involving a

zone of inhibition to qualitatively observe the decrease in bacterial concentration. The zone of

inhibition is simply the area on the agar plate that remains free from microbial growth. Zones of

inhibition are used to measure how potent an antimicrobial agent is against bacteria. A paper disc

that is saturated with the antimicrobial agent is placed on an agar surface that has been streaked

with bacteria. If substantial antimicrobial activity is present after a set amount of days depending

on the bacteria grown, a clear region will form around the disc, termed the “zone of inhibition”.

The zone of inhibition around the test product indicates the absence of microbial growth by the

antimicrobial agent. This method of testing is fast and well-suited for determining the ability of

water-soluble antimicrobials to inhibit the growth of a number of sample microorganisms.

A toothbrush may not be handy at the holiday dinner table, but research published in the

journal Food Chemistry suggested moderate consumption of red wine helps to rinse teeth clean

of bacteria during and after meals (Discovery News, 2009). The case study is an investigation

about how red wine may prevent cavities, specifically involving proanthocyanidins (from the red
wine) and their effect on Streptococcus mutans (S. mutans). The findings add to the growing list

of health benefits associated with drinking wine, including improved longevity and diminished

risk of cardiovascular and neurological diseases. Although alcohol's cavity-preventing benefits

are already well known, we became interested in the dental benefits for teetotalers. The

researchers cultured S. mutans, common bacteria that feed on sugars in food and contribute to

tooth enamel demineralization, which often results in cavities. Streptococcus is a group of

spherical bacteria that belong to a group termed Streptococcaeceae, which may cause different

diseases in humans and animals. Streptococcus mutans (S. mutans), specifically, belong to the

viridians species which thrive in the mouth and contribute to tooth decay. S. mutans cause tooth

decay by metabolizing simple sugars and fermentable carbohydrates into acids that dissolve

tooth enamel. Many factors including diet, general health, and heredity affect an individual’s

chance of developing caries, or tooth decay. We initially planned to simulate a mouth to test the

effect of non-alcoholic wines in reducing the concentration of S. mutans on calcium ceramic

beads using water and LB agar as a bacterial growth medium. However, we were unable to

obtain S. mutans, so we had to revise our experiment and substitute S. mutans with E. coli.

Consequently, based on recent outbreaks of E. coli food poisoning, we decided the main focus of

our project be on the function of different types of wine as a disinfectant.