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(Solved) Lab 6 - Experiment 1: Winogradsky Column Sergei Winogradsky


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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.

 

Materials:

 

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2 Full pages of shredded newspaper

 

(2) 5 g. Bags of calcium carbonate (chalk)

 

(2) 5 g. Bags of calcium sulfate

 

Paint stirrer

 

Parafilm?

 

(2) Rubber bands

 

Permanent marker

 

Ruler

 

1 pair of gloves

 

4 Large plastic bags

 

250 mL Beaker

 

Trowel

 

*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

 

Procedure:

 

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?

 


 

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? 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

 

this procedure.

 

? 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!

 

i.

 


 

Completely submerge the 250 mL beaker in the water body, being

 

careful to avoid collecting as much sediment and debris as possible.

 

ii.

 

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).

 

iii.

 

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.

 

i.

 

Remove any surface debris (plant residues/leaves/branches/ thatch/etc.)

 

from the soil site.

 

ii.

 

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

 

zone.

 

? Collect in between crop rows if collecting from a fertilizer band.

 

? Try to sample dark, light, limed, and unlimed soil areas separately.

 

iii.

 

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)

 

iv.

 

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.

 


 

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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

 

air.

 

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

 

Week #

 


 

Observations (Colors, Layers, Observations (Colors, Layers,

 

Column Location) of Column in Column Location) of Column in

 

Light

 

Dark

 


 

1

 

2

 

3

 

4

 

5

 

6

 

Questions:

 

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?

 


 

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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

 

fittest??

 


 

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