WaterWater

By:By:

Andrew PaekAndrew Paek

Nirav ParekhNirav Parekh

BackgroundBackground

Water is an essential part of our well beingWater is an essential part of our well being We use it for nourishment, cooking, We use it for nourishment, cooking,

cleansing, and many other things.cleansing, and many other things. Because of water’s importance, there are Because of water’s importance, there are

methods used to purity water to make safe methods used to purity water to make safe for human consumption.for human consumption.

Federal regulations are put in place to Federal regulations are put in place to ensure this safety.ensure this safety.

PurificationPurification

To purify water…To purify water… 1) Raw water (untreated water) is pumped into a 1) Raw water (untreated water) is pumped into a

sediment basin where large particles such as sand sediment basin where large particles such as sand settle to the bottom.settle to the bottom.

2) The sediment free water is then pumped into the 2) The sediment free water is then pumped into the coagulation basin where the anionic polymers and coagulation basin where the anionic polymers and chlorine added to the sediment basin, which helps chlorine added to the sediment basin, which helps form larger insoluble particles out of the suspended form larger insoluble particles out of the suspended tiny particles in the water.tiny particles in the water.

3) The water is filtered to remove remaining solutes, 3) The water is filtered to remove remaining solutes, suspended particles and microorganismssuspended particles and microorganisms

DisinfectionDisinfection

Filtered water must be disinfected before it Filtered water must be disinfected before it is released as drinking wateris released as drinking water

Chlorine is the most common disinfectantChlorine is the most common disinfectant Kills microorganisms within 30 minutesKills microorganisms within 30 minutes Neutralizes organic compoundsNeutralizes organic compounds Improves water taste and smellImproves water taste and smell

RegulationsRegulations

There are certain bacteria known as coliforms There are certain bacteria known as coliforms which are used to indicate if water is which are used to indicate if water is contaminated.contaminated.

Based on the Based on the Safe Drinking Water ActSafe Drinking Water Act, water is , water is only considered safe if the average number of only considered safe if the average number of coliform bacteria does not exceed 1 CFU/100 coliform bacteria does not exceed 1 CFU/100 mL mL

Water utilities report results to the EPA if they Water utilities report results to the EPA if they don’t meet standardsdon’t meet standards

The public is also informedThe public is also informed Outbreaks are reported to the CDCOutbreaks are reported to the CDC

HypothesisHypothesis

We predict that water from various drinking sites We predict that water from various drinking sites on campus on campus will will meet federal regulations. As of meet federal regulations. As of now, there isn’t any kind of public now, there isn’t any kind of public announcement that the water on campus isn’t announcement that the water on campus isn’t safe to drink, so it can be assumed that the safe to drink, so it can be assumed that the water on campus meets federal drinking water on campus meets federal drinking regulations.regulations.

We predict that we will not find any coliforms in We predict that we will not find any coliforms in the water samples intended for drinking.the water samples intended for drinking.

MethodsMethods

Obtained three 100 mL samples of water Obtained three 100 mL samples of water at five sites on this campus:at five sites on this campus: Dorm water fountainDorm water fountain Dorm showerDorm shower Dorm sinkDorm sink Outdoor fountainOutdoor fountain Diner drinking waterDiner drinking water

These samples were filtered, incubated, These samples were filtered, incubated, and the number of colonies were countedand the number of colonies were counted

Membrane Filtration MethodMembrane Filtration Method

1) Shake each 100ml sample in their 1) Shake each 100ml sample in their containers at least 25 times.containers at least 25 times.

2) Pour 100ml of each sample into the 2) Pour 100ml of each sample into the filter apparatus.filter apparatus.

3) Allow the water to filter for at least an 3) Allow the water to filter for at least an hour.hour.

4) Pour each of the 2ml ampules of the m-4) Pour each of the 2ml ampules of the m-coliblue24 broth into each sample.coliblue24 broth into each sample.

Rational for MF methodRational for MF method

The m-coliblue24 broth is a type of media The m-coliblue24 broth is a type of media that selects for coliforms. It’s also a that selects for coliforms. It’s also a differentiatial media which detects differentiatial media which detects between E.Coli and other coliforms.between E.Coli and other coliforms.

The membrane filtration method allows us The membrane filtration method allows us to count the number of colony forming to count the number of colony forming units which allows us to determine if the units which allows us to determine if the water samples meet federal regulations.water samples meet federal regulations.

Plating and EnterotubesPlating and Enterotubes

We inoculated the Eosin-methylene blue We inoculated the Eosin-methylene blue plates with unique colonies found from the plates with unique colonies found from the filters.filters.

From the pure colonies we inocoluated From the pure colonies we inocoluated enterotubes which tests for various enterotubes which tests for various characteristics of the colonies found in the characteristics of the colonies found in the water samples.water samples.

These methods are used to identify the These methods are used to identify the coliforms found in the water samples.coliforms found in the water samples.

Expected ResultsExpected Results

For the MF technique:For the MF technique: Total coliforms would appear as red colonies.Total coliforms would appear as red colonies. E.coli would appear as blue colonies.E.coli would appear as blue colonies.

For the eosin-methylene blue media:For the eosin-methylene blue media: Only coliforms should be able to grow.Only coliforms should be able to grow. Lactose fermenters such as E.coli would Lactose fermenters such as E.coli would

appear with a green sheen.appear with a green sheen.

ResultsResults

No coliforms were detected in these water No coliforms were detected in these water samples:samples: Dorm water fountainDorm water fountain Dorm showerDorm shower Dorm sinkDorm sink Diner drinking waterDiner drinking water

Colonies were found from the outdoor Colonies were found from the outdoor fountain.fountain.

Filters from the Outdoor fountainFilters from the Outdoor fountain

Filter 1Filter 1

5 brown and 3 5 brown and 3 pink/purple pink/purple coloniescolonies

Filter 2Filter 2

1 brown and 5 1 brown and 5 pink/purple pink/purple coloniescolonies

About 80 tiny About 80 tiny purple colonies.purple colonies.

Filter 3Filter 3

No colonies No colonies observed.observed.

Filter 2Filter 2

EMB PlatesEMB Plates

Outdoor Filter 1Outdoor Filter 1 Control E.ColiControl E.Coli

Enterotube ResultsEnterotube ResultsControl: e.coli Outdoor Filter 1 colony

Glucose +gas -Lysine + -Ornithine - -

H2S - -

Indole + -Donitol - -Lactose + -Arabinose + -Sorbitol + -Voges Proskauer - -Dulcitol - -PA - -Urea - -Citrate - +

AnalysisAnalysis

Since no coliforms were present in the sites Since no coliforms were present in the sites where people drink water, we can conclude that where people drink water, we can conclude that the drinking water here on campus is safe the drinking water here on campus is safe according to federal regulations.according to federal regulations.

DON’T DRINK WATER FROM THE OUTDOOR DON’T DRINK WATER FROM THE OUTDOOR FOUNTAINS.FOUNTAINS.

The isolated colonies found on the filter could The isolated colonies found on the filter could possibly be from the genus possibly be from the genus Pseudomonas, Pseudomonas, which doesn’t produce acids or gas from which doesn’t produce acids or gas from glucose. These are normally found in soil and glucose. These are normally found in soil and water.water.

Implications of coliformsImplications of coliforms

We’re interested in finding bacteria called We’re interested in finding bacteria called coliformscoliforms, which are facultatively aerobic, gram-, which are facultatively aerobic, gram-negative, nonspore-forming, rod-shaped negative, nonspore-forming, rod-shaped bacteria which are able to ferment lactose and bacteria which are able to ferment lactose and produce gas.produce gas.

Their presence could indicate that the water Their presence could indicate that the water contains fecal matter.contains fecal matter.

Coliforms are not necessarily responsible for Coliforms are not necessarily responsible for serious illnesses .serious illnesses .

If the water is dirty enough to have fecal matter, If the water is dirty enough to have fecal matter, chances are that there are pathogens in the chances are that there are pathogens in the water as well.water as well.

Implications (cont.)Implications (cont.)

Some of the main microorganisms which Some of the main microorganisms which cause serious waterborne illnesses:cause serious waterborne illnesses: Typhoid fever – Salmonella typhiTyphoid fever – Salmonella typhi Cholera – Vibrio choleraeCholera – Vibrio cholerae Legionellosis – Legionella pneumophilaLegionellosis – Legionella pneumophila Giardiasis – Giardia IntestinalisGiardiasis – Giardia Intestinalis Amebiasis – Entamoeba histolyticaAmebiasis – Entamoeba histolytica

Frequency of disease in the USFrequency of disease in the US

In the year 2001-2002 there were 6 outbreaks In the year 2001-2002 there were 6 outbreaks caused by the bacteria Legionella which resulted caused by the bacteria Legionella which resulted in:in: Illnesses in 80 peopleIllnesses in 80 people 41 hospitalizations41 hospitalizations Four deathsFour deaths

This was the most prevalent cause of This was the most prevalent cause of waterborne illness out of the other causes waterborne illness out of the other causes including chemicals, parasites, viruses, and including chemicals, parasites, viruses, and other bacteria.other bacteria.

LegionellosisLegionellosis An intracellular parasite that grows in alveolar An intracellular parasite that grows in alveolar

macrophages and monocytes. macrophages and monocytes. Leads to pneumonia or a mild infection known as Leads to pneumonia or a mild infection known as

Pontiac feverPontiac fever Symptoms:Symptoms:

High feverHigh fever CoughCough Muscle aches and headachesMuscle aches and headaches

TreatmentTreatment Antibiotics – rifampin and erythromycin Antibiotics – rifampin and erythromycin

PreventionPrevention The bacteria can be killed in the water supply through The bacteria can be killed in the water supply through

hyperchlorination or heating the water to temperatures greater hyperchlorination or heating the water to temperatures greater than 65 degrees Celsius.than 65 degrees Celsius.

Protocol EvaluationProtocol Evaluation

Water samples were Water samples were not dechlorinated not dechlorinated which could have which could have inhibited growth.inhibited growth.

As seen on the As seen on the control, some bacteria control, some bacteria may not have grown may not have grown on raised parts of the on raised parts of the filter.filter.

Comments on the resultsComments on the results

In the enterotube that tested the sample from the In the enterotube that tested the sample from the outdoor fountain, only the last test was positive. This is outdoor fountain, only the last test was positive. This is very unusual since coliforms should be able to ferment very unusual since coliforms should be able to ferment some kind of sugar being facultative anerobes. This some kind of sugar being facultative anerobes. This colony should be tested again for identification.colony should be tested again for identification.

Only two out of six colonies (including the control) from Only two out of six colonies (including the control) from the filter membranes were able to grow on the EMB the filter membranes were able to grow on the EMB agar. The process which the m-coliblue24 broth and the agar. The process which the m-coliblue24 broth and the EMB agar select for coliforms may be different between EMB agar select for coliforms may be different between each other. This would cause one media to select each other. This would cause one media to select against some colonies which may have grown in the against some colonies which may have grown in the other media.other media.

Overall significance of the ProjectOverall significance of the Project

Based on our tests on the water samples Based on our tests on the water samples across campus, we can conclude that the across campus, we can conclude that the water is safe to drink.water is safe to drink.

We can also conclude that water in We can also conclude that water in outdoor fountains contain fecal matter. So outdoor fountains contain fecal matter. So it’s advised that nobody drinks or touches it’s advised that nobody drinks or touches it.it.

THE ENDTHE END

Bibliography Bibliography Madigan, Michael and Martinko, John. Madigan, Michael and Martinko, John. Brock Brock

Biology of Microorganisms. Biology of Microorganisms. Upper Saddle River, Upper Saddle River, NJ: Pearson Prentice Hall. 2006NJ: Pearson Prentice Hall. 2006

Centers for Disease Control and Prevention. Centers for Disease Control and Prevention. Surveillance for waterborne-disease outbreaks Surveillance for waterborne-disease outbreaks associated with recreational water — United associated with recreational water — United States, 001–2002 and Surveillance for States, 001–2002 and Surveillance for waterbornedisease outbreaks associated with waterbornedisease outbreaks associated with drinking water — United States, 2001–2002. In: drinking water — United States, 2001–2002. In: Surveillance Summaries, October 22, 2004. Surveillance Summaries, October 22, 2004. MMWR 2004:53(No. SS-8).MMWR 2004:53(No. SS-8).