Lab #5 A Succession of Microbes Introduction: In natural communities, the replacement of species that occurs over time is called succession.

. It may be caused by several different factors, including competition, changing resources, or one species creating resources for species that follow. The newer species thrice and may dominate the community, but they are eventually replaced by still other species until a stable climax community forms. Succession is especially noticeable after natural disasters or severe disturbances caused by humans. In this investigation, you will study succession in two communities of microscopic organisms living in wet soil. You will place each of your communities in a Winogradsky column, a device that was originally developed by Russian microbiologist Sergei N. Winodradsky in the 1880s. You will add various nutrients to one of the columns. The other column will be your control, and you will not ass anything to it. By comparing the experimental Winogradsky column with the control, you will study the effects of added nutrients on microbial succession. Materials: 2 clear plastic, 500 ml soda bottles with screw on caps. 1000 cubic centimeters of soil from a natural area Dowel Funnel (large bore hole) Spoon Fresh water from a naturally occurring source (pond) Shredded newspaper of a crushed egg shell of a crumbled, hard-boiled egg yolk Metric ruler Wax pencil or permanent marker Procedure: 1. Create a data sheet to collect data once a week for the next 10-12 weeks. Keep in mind that you will be looking at the environment as a whole and at the microbes growing in your environment. 2. Collect about 1000 cu. cm of soil from a natural source. Collect several hundred milliliters of water from a pond. Leave them in a sunny window or under an artificial grow light over night to settle. 3. Build the Winogradsky columns. a. Remove any hard lumps, twigs, rocks, and other large particles from about 1000 cu. cm of soil. Separate the soil into two equal piles. b. Add a small handful of well-shredded newspaper, a crushed egg shell, and about of a crumbled yolk from a hardboiled egg to one of the piles of soil. These added substances will act as nutrients. Mix the nutrients thoroughly with the soil. c. Remove the product labels from the bottles. Label the two plastic bottles, one as the control, and one as the experimental bottle. d. Using the funnel, gently pour the soil that contains the added nutrients into the experimental bottle to a height of about 5 cm. Then gently pour the soil without the added nutrients into the control bottle to a height of about 5 cm. Use the dowel to pack the soil into the bottom of each bottle and to remove the air trapped in the soil. Add a little more soil and a few spoonfuls of pond water to each bottle, using the dowel again to pack the soil and release any trapped air. Repeat this process until the level of the spoil is about 5 cm from the top of each bottle, and cap the bottles tightly. e. Place your Winogradsky columns in an area where they will receive indirect sunlight. 4. Make sketches and take pictures of the columns and their contents once a week for 10-12 weeks. You may observe your bottle more than once a week if you choose. Do not open the columns until you reach the end of the 6 weeks. Record your observations in your data sheet. 5. Use the following key to describe the succession of microbe populations that you observed in the bottles over the course of the six weeks: a. Green clumps: algae b. Black spots: sulfate-reducing bacteria c. Reddish purple spots: purple sulfur bacteria d. Rust-colored areas: purple non-sulfur bacteria e. Any other colored spots, create a key of your own. Describe any other populations you may have observed.

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Write a lab report for your experiment, due 13 weeks after completion of step 3. In your conclusion, answer the following questions: a. What differences did you observe between the two columns in terms of the rate and pattern of succession? b. A simple sulfur cycle is going on in the columns. The sulfate-reducing bacteria produce a substance called hydrogen sulfide (the black spots), which is used by the purple sulfur bacteria for photosynthesis. The purple sulfur bacteria then produce other forms of sulfur, which are used by the sulfur-reducing bacteria. Create a simple pictorial cycle to show what is happening in the column. Given this information, what could you do to stop this cycle? c. What could have caused the succession in your bottle, and how is it different from the succession in a natural ecosystem? d. What might have happened if you had chosen soil from a different source, such as a beach, your back yard, or near the sewer processing plant? e. How does this lab differ from your pond succession lab (lab #4)?