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Lab 6 - Experiment 1: Winogradsky Column
Sergei Winogradsky (1856-1953) was a Russian microbiologist who was one of the first
scientists to study mixed populations of bacteria with differing metabolic abilities. In a
Winogradsky column, two distinct gradients form in opposite directions: oxygen is high at the
top of the column and non-existent at the bottom while hydrogen sulfide is high at bottom and
non-existent at the top. Additionally, light facilitates the growth of aerobic photosynthetic
bacteria. These generated gradients allow for growth of a large variety of bacteria that
reproduce the biogeochemical cycles on which life depends.
2 Full pages of shredded newspaper
(2) 5 g. Bags of calcium carbonate (chalk)
(2) 5 g. Bags of calcium sulfate
(2) Rubber bands
1 pair of gloves
4 Large plastic bags
250 mL Beaker
*2 Clean, clear 2 L plastic bottles with the top cut off
*8 - 10 C. of dirt or mud (pond, lake, city park, yard, etc.)
*8 - 10 C. of water (preferably collected from the same location as the dirt/mud); tap
water will also suffice).
? *Light source (south facing window or lamp with no more than a 60 W bulb)
? *Large bowl
*You must provide
Field Trip - Water and Soil Sample Collection
1. Identify a location that has water and soil available for collecting; and, which you believe
has features abundant micro-organismic, plant or animal life. Keep in mind that you will
need 8 - 10 cups of water and 8 - 10 cups of soil.
2. Use graph paper to sketch a diagram of where you will collect your water and soil
samples from. Be sure to label the graph with descriptive information, and indicate any
other critical information that might affect the water quality. For example:
? Has it recently flooded?
? Is the area experiencing a drought?
? Is there snow or ice present?
? What is the temperature?
? Is the surrounding animal/marine life steady/predictable?
? What kind of animals/plants are local to the area?
? Is it humid out?
This diagram is called a Field Report and will become useful should you need to
re-assess the original environment.
3. Label each plastic bag with the location name and the collection date.
4. Go into ?the field? to obtain your samples. To do this:
? Pack up your plastic bags, gloves, ruler, 250 mL Beaker, and a printed copy of
? Water sample collection: (you can collect your soil sample first; refer to the
bottom half of this section for soil sample collection methods).
NOTE: Some collectors may wish to wade into the water to obtain their water
sample. As a general rule of thumb, water collectors should NOT wade into
flowing water if the water depth (in feet) x the water velocity (in feet per second)
is greater than or equal to 10. If you have any hesitations regarding water-safety,
do NOT enter the water!
Completely submerge the 250 mL beaker in the water body, being
careful to avoid collecting as much sediment and debris as possible.
Transfer the water from the beaker into one of the plastic bags designated
for your water sample. Repeat this process 5 times for
each bag (a total of 10 times).
Seal the bags tightly and store them away or set aside until you
leave the area.
? Collect your soil sample. It is ideal to use the same environment that the
water sample came from. However, you can move to a new location if necessary.
Remove any surface debris (plant residues/leaves/branches/ thatch/etc.)
from the soil site.
Insert your trowel into the soil and observe the following depth guidelines
depending on the soil environment:
? Sample the soil down about 6 - 8 in. deep if collecting from a
garden/ flower bed zone (use your ruler to approximate the depth).
? Sample the soil down about 3 in. deep if collecting from a turf zone.
? Sample the soil down about 8 - 12 in. deep if collecting from a root
? Collect in between crop rows if collecting from a fertilizer band.
? Try to sample dark, light, limed, and unlimed soil areas separately.
Scoop up the soil, and transfer it into one of the plastic bags designated
for the soil. The trowel in your lab kit can contain 1/3 of a cup. Therefore,
you will need to repeat this process 12 -15 times for each plastic bag (a
total of 24 - 30 times)
Try to return back to your ?lab? without changing the structure of the soil
composition. Natural clumps, rocks, etc. should be maintained to get a
more authentic understanding of the soil porosity and biochemical habitat.
2. Put approximately 946 mL (4 C.) of the dirt/mud into a large bowl or bucket, add 1 bag
(5 g.) of the calcium carbonate, and 1 bag (5 g.) of the calcium sulfate to the mixture.
Note: There are 236 mL in one cup.
3. Add enough of the water you collected to make a thick, but somewhat fluid, mixture.
This should require approximately 946 mL (4 C.).
4. Add approximately 1/2 of the shredded newspaper to the bowl and mix again.
5. Transfer this mixture to the soda bottle and tap the bottle on the ground or other hard
surface to pack the mixture tightly to the bottom. It is important that no air pockets or
bubbles should remain in this layer
6. Use a paint stirrer or handle of a long spoon to further pack the mud and remove any
7. Add approximately 236 mL (1 C.) of the remaining (unmodified) dirt/mud on top of the
previous layer and tap again to pack it down.
8. Add water to a depth of approximately 1 in. above the last layer and make a small mark
with a permanent marker on the 2 L bottle at the top of the water level.
9. Let the column sit undisturbed for 30 mins. and monitor the depth of the water. If the
water level rises, remove some to return to the original level. If the water level
decreases, add more to return to the original level. There should be approximately 1 in.
of air space above the water.
10. Cover the top with Parafilm? and secure with a rubber band.
11. Repeat the Steps 5 - 13 to create a second Winogradsky column.
12. Incubate the columns for 6 - 10 weeks at room temperature. Place one column near a
south-facing window to receive indirect sunlight, or approximately 2 ft. away from a lamp
with a 40 - 60 W. bulb. Place the second column in a dark location (without natural or
artificial light available). Remember to rotate the columns 180? 1 time per week.
13. Observe the column every 7 days and record your observations in Table 1.
Table 1: Winogradsky Column Observations
Observations (Colors, Layers, Observations (Colors, Layers,
Column Location) of Column in Column Location) of Column in
1. Do ecosystems change over time? If so, what causes those changes to occur?
2. How did your results vary between the two columns? Why do you think their responses
were different? Be specific.
3. Where does carbon come from in a Winogradsky column?
4. What is carbon important?
5. What purpose does calcium sulfate serve in the Winogradsky column?
6. How is photosynthesis different between cyanobacteria (growing at the top of the column
and green and purple sulfur bacteria (growing near the bottom 1/3 of the column)?
7. Identify three critical factors (abiotic or biotic) required for primary succession to occur.
8. Define two reasons why ecological succession progresses from populations with low
diversity to populations with high diversity.
9. Highly diverse ecosystems are often regarded as a more ?healthy? ecosystem. Explain why.
10. How does ecological succession demonstrate the evolutionary process of ?survivial of the
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