Name _________________________

Exercise 6:
A Trip to the Sewage
Treatment Plant Sample
Collection and Quantitative
Analysis

INTRODUCTION
In this exercise we will assess the effect of Bard's drinking water plant on its input
(water from the Sawkill) and the effect of the sewage treatment plant on its input (raw
sewage). To do this, we will collect water samples from the input and output of each
plant and we will determine the concentration of bacteria in each. The four samples in
mind are quite instructive, but you can take more if you are curious about anything in
particular.

PROCEDURE
1.

Obtain four autoclaved milk dilution bottles. Label them

SAWKILL INPUT
DRINKING WATER OUTPUT
RAW SEWAGE INPUT
TREATED SEWAGE OUTPUT

2.

After this we will head to the Bard College drinking water and sewage treatment
plants to take tours and to collect samples. Bring plastic gloves and a strong
stomach for the latter.

3.

After the tours bring the samples back to lab to determine the concentration of
bacteria in each sample. To do this, we will prepare dilutions of each of samples.
This will involve making a series of 10-fold dilutions and will be described in detail
in the lab. We will make 100 (undiluted), 101, 102, 103, 104, 105, 106, and
107 dilutions of each.

4.

After the dilutions are made, pipette 1.0 mL of each dilution (NO MOUTH
PIPETTING ) into separate screw-cap culture tubes containing molten agar at

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EXERCISE 6: A TRIP TO THE SEWAGE TREATMENT PLANT
47 C. Mix each 47 C tube swiftly but thoroughly, without making any bubbles,
then immediately pour the agar into the bottom of a sterile Petri dish. The technique will be demonstrated. KEEP THE 47 C TUBES OUT OF THE 47 C
WATER BATH FOR ABSOLUTELY AS LITTLE TIME AS POSSIBLE, OR
THEY WILL GEL ON YOU!
5.

Incubate the plates at room temperature until colonies have grown sufficiently for
counting. The colonies confined in the agar will be much smaller than those
growing freely on the agar. Since all colonies are presumably the product of a
single cell, count them all equally, no matter what the size.

RESULTS
1.

For each water sample, find a plate that has between 30 and 300 colonies, count
the colonies and record the number below. Indicate the dilution of the sample
used for each plate counted. Use this data to calculate how many bacteria there
were for each sample per milliliter before diluting.

Water Sample

Colonies per
Plate

Cells per mL
of Dilution

Dilution
Factor

Cells per mL
of Original
Sample

Sawkill Input
Drinking Water
Output
Raw Sewage
Input
Treated
Sewage Output

DISCUSSION
1.

Was the concentration of bacteria in the Sawkill input to the drinking water plant
higher than that of the drinking water output?

What procedures in the drinking water plant are designed to reduce the bacterial
count?

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EXERCISE 6: A TRIP TO THE SEWAGE TREATMENT PLANT
2.

Was the concentration of bacteria in the treated sewage output lower than that of
the raw sewage input?

What procedures in the sewage treatment plant are designed to reduce the
bacterial count?

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EXERCISE 6: A TRIP TO THE SEWAGE TREATMENT PLANT

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