1. Carolina Science Supply

2.  The activities in this kit are intended to simulate how oil-degrading microbes can be used to break down petroleum after oceanic oil spills.  The activities are NOT a direct representation of marine oil spill bioremediation.

3.  Carolina Item #181322  $89.25  Kit designed for a class of 32 students

4.  Included in the kit:  1 bottle of tetrazolium indicator, 0.02% ▪ Chemical indicator that will turn from clear to pink when reduced  1 bottle of oil  1 container of Rid-X Septic System Treatment  1 sheet of labels  64 culture tubes with caps  81 plastic pipettes  Needed, but not supplied:  Test tube racks  At lease 64 mL of distilled water  140 mL warm tap water  Funnel  20 x 20 Cheese cloth

5.  Prior to the lab, the Rid-X solution needs to be made, left to react, and then sifted.  This process can be completed the day before the lab.

6.  In a spill of crude oil in the ocean:  25% of the oil evaporates and becomes air pollution  15% of the oil consists of heavy compounds which stick together and sink to the bottom of the ocean  The remaining 60% is broken down over time into simpler, nontoxic products by oil-degrading microbes (microscopic living organisms) in a process called biodegradation.

7.  Human intervention to facilitate microbial removal of pollutants from the environment  Three major approaches to bioremediation for the cleanup of marine oil spills

8.  Biostimulation  Optimizing the environment of oil-degrading microbes to accelerate their growth and metabolic activity  Nutrients such as nitrogen, phosphorus, and iron are added to the ocean environment, much like applying fertilizer to a lawn  Negatives ▪ Nutrients must remain in contact with the oil ▪ Concentration must remain at an optimal level for an extended period of time.

9.  Bioaugmentation  The process of supplementing (or “seeding”) a population of naturally occurring, oil-degrading microbes with additional microorganisms  Used when the existing population of microbes in a contaminated region is not optimally suited to degrade the type of oil present  Positives ▪ These microbes cans be easily cultivated in large quantities in laboratories and can be stored, ready for use, for up to three years

10.  Genetically Engineered Microorganisms (GEM)  Most naturally occurring microbes cannot achieve complete degradation on its own  Scientists are creating genetically modified microorganisms to combine requisite pathways and enzymes in one organism in order to degrade oil more efficiently and extensively  Relatively new approach and is undergoing development

11.  Lab consists of four culture tubes to be used in two experiments Experiment A ▪ Oil will be treated with oil-degrading microbes (experimental condition) and without oil-degrading microbes (control condition) in the presence of the tetrazolium chemical indicator Experiment B ▪ Oil will be treated with oil-degrading microbes (experimental condition) and without oil-degrading microbes (control condition) but without the chemical indicator