The Effect of Fertilizers on Microbial Survivorship Jonah Bruemmer 9th Grade

Introduction Many different types of fertilizers are used to increase plant growth. This fertilizer can leak into groundwater, and the nutrients from the fertilizer cause overgrowth of algae. Algae population consumes the supply of dissolved oxygen and organisms no longer have oxygen to live, which is a problem. Fertilizers contaminating groundwater is a problem. Can fertilizers affect environmental prokaryotic populations?

Chemical Fertilizer Miracle-Gro Liquid All Purpose Plant Food Liquid form Advised to be used diluted with water 29.6ml of fertilizer/3785ml of water (1 gallon) 12-4-8 (nitrogen %- phosphorus %- potassium %) Not organic

Biological Fertilizer Alaska Fish Fertilizer Liquid form Advised to use at 29.6ml/3785ml of water (1 gallon) 5-1-1 (nitrogen %- phosphorus %- potassium %) Organic

Escherichia Coli (E. coli) Found in intestinal tract of animals Contaminates groundwater through feces Mostly harmless except for a few strains Prokaryotic Gram negative

Purpose To examine the effect of a biological and a chemical fertilizer on E. coli survivorship.

Hypotheses Null: The different concentrations of the two fertilizers will not have an effect on the survivorship of E. coli cell colonies. Alternate: The different concentrations of the two fertilizers will have an effect on the survivorship of E.coli cell colonies. The chemical fertilizer will have a greater effect than the biological fertilizer.

Materials MiracleGro Chemical Fertilizer. Alaska Fish Biological Fertilizer LB Media (1% tryptone, 5% yeast, 10% NaCl) LB media + 12% agar Plates E. coli (DH5Alpha) Incubator at 37°C Pipette (p. 20, 200, 2000) Test tubes Spreader bars Vortex Sharpie Sterile Filter Syringes

Methods Both fertilizers were sterile filtered and then diluted with water to their recommended application. (30ml/3785ml) Both fertilizers were pipetted into tubes with LB media according to their desired concentration (0%, 1%, 2%, 5%, 10%)

Experimental Concentrations(per fertilizer) 0% (control) 1% 2% 5% 10% ml of E. coli 0.1ml ml of LB media 0.2ml 0.197ml 0.194ml 0.185ml 0.170ml ml of fertilizer 0ml 0.003ml 0.006ml 0.015ml 0.03ml Total Amount Per Tube 0.3ml

Methods (Continued) 0.1ml of E. Coli cells (10^5) were added into each tube 0.1ml samples from each tube was pipetted onto LB plates (5 plates per concentration) Plates were allowed to incubate at room temperature for 10 minutes Plates were incubated for 36 hours at 37°C and counted visually

MiracleGro Chemical Fertilizer Effects on E. coli Survivorship p value = 0.00035 Concentrations of MiracleGro Application Stock

Dunnett’s Test - MiracleGro Concentration T-value Result 1% 2.19 Insignificant 2% 3.88 Significant 5% 4.31 10% 5.20 T-crit = 3.48

Alaska Fish Fertilizer Effects on E. Coli Survivorship p value = 0.0000003 Concentrations of Alaska Fish Fertilizer Application Stock

Dunnett’s Test - Alaska Fish Fertilizer Concentration T-value Result 1% 1.60 Insignificant 2% 3.35 5% 4.29 Significant 10% 8.90 T-crit = 3.48

MiracleGro and Alaska Fish Fertilizer Effect on E. coli Survivorship

Conclusions As concentrations of fertilizer increased, the amount of E. Coli colonies increased as well. The null hypothesis can be rejected for both fertilizers for the 5% and 10% concentrations and the 2% concentration of MiracleGro The null hypothesis cannot be rejected for both of the fertilizers for both 1% concentrations and the 2% concentration of Alaska Fish Fertilizer only. Compared to the control, higher concentrations of Alaska Fish Fertilizer increased the amount of colonies much more than MiracleGro.

Limitations Only one type of bacteria was tested Only 5 Concentrations Only two fertilizers were tested Plating techniques could be unsynchronized (varying time of exposure) Study does not reveal other effects on the bacteria

Extensions Test more than one bacteria type Test more than two fertilizers More concentrations with more trials Different methods of testing (agar infusion) Test only nitrogen

Sources “E. Coli (Escherichia Coli) | E. Coli | CDC.” Centers for Disease Control and Prevention, Centers for Disease Control and Prevention, www.cdc.gov/ecoli/index.html. Perlman, Howard, and Usgs. “Nitrogen and Water.” Adhesion and Cohesion Water Properties, USGS Water Science School, water.usgs.gov/edu/nitrogen.html.

ANOVA : MiracleGro SUMMARY Groups Count Sum Average Variance 5 222 5 222 44.4 17.8 1 253 50.6 5.3 2 277 55.4 12.8 283 56.6 23.8 10 296 59.2 40.7 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 682.96 4 170.74 8.502988048 0.0003536327302 2.866081402 Within Groups 401.6 20 20.08 Total 1084.56 24

ANOVA : Alaska Fish Fertilizer SUMMARY Groups Count Sum Average Variance 5 120 24 57.5 1 154 30.8 22.7 2 191 38.2 50.2 211 42.2 41.7 10 310 62 53.5 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 4155.76 4 1038.94 23.02615248 0.0000003010269956 2.866081402 Within Groups 902.4 20 45.12 Total 5058.16