The effect of water on bacteria survival By Declan Reilly

Introduction In this day and age, water can be bought in many different forms and packaging. For example, some types of water have different products such as minerals or electrolytes added to them. These minerals and electrolytes can foster a good environment for bacteria to live and flourish, which can become a major health risk. 

Variable Smart water Has electrolytes that improve your energy. Has other minerals that help with taste. This will most likely wield the greatest results from the bacteria. Contains vapor distilled water one liter, calcium chloride 0.2%, magnesium chloride 0.9% and potassium bicarbonate 0.4% electrolyte 1% for taste.

Regular(Kirkland ) bottle water It is predicted there will be a low amount of bacteria surviving colonies from this type of water. purified water potassium bicarbonate 0.1% calcium citrate 3% magnesium oxide 2% calorie free sodium free

Tap The tap water will have a low survival rate, tap water is tested by Pittsburgh Water & Sewer Authority for a bacteria or disease. Because chlorine is used to treat the water, there may be some chlorine residue remaining. This will help to kill bacteria leading to the low surviving colonies. Fluoride- 2ppm Nitrate- 3ppm Barium- 1ppm Cyanide- 0.2ppm Total organic compound- 39.2ppm

Tap continued Trihalomethanes- 13ppm Haloacetic Acids - 8ppm NaCEL- 21ppm Free Chlorine- 0.2ppm

MODEL BACTERIA E. coli Prokaryotic, gram negative, unicellular, bacterial cell  Intestinal mammalian symbiont  Inexpensive laboratory variable, easily cultured Pathogen

SDF Isotonic environment Used as a controlled variable for many experimented. SDF (Sterile Dilution Fluid) 100mM KH2P04 ,10mM MgSO4, 1mM NaCl

Purpose If the bacteria can survive and thrive this can lead to a major health problem.

Hypothesis Alternative hypothesis: The different kinds of bottled water, such as the Smart Water will have an increase of bacteria survival of the colonies. Null hypothesis: The different kinds of bottle water will have no effect on the survival of the bacteria .

Materials •Sidearm Flask •Turn-table Spreader bars •Vortex •Incubator •Cone/Beaker •Tap water, Smart water, regular bottle water •SDF (Sterile Dilution Fluid)- 100mm KH2PO4, 10mm MgSO4, 1mm NaCl •Ethanol Spreader bars YEPD Agar Plates (1% Yeast Extract, 2% Peptone, 2% Glucose) Escherichia coli C600 •Burner

Procedure 1. E. coli was grown overnight in sterile LB media. 2. A sample of this culture was added to LB media in a side arm flask. 3. The culture was incubated until a density of 50 Klett units.

Concentration grid Type SDF Tap Smart water Regular Water E. Coli 0.1ml Variable 9.9ml Total 10ml Concentration 99%

Procedure 4. 0.1mL of cell culture was then added to the test tubes, yielding a final volume of 10mL and had a cell density of approximately 10^3 cells/mL/ 5. The solution was mixed by vortex. 6. After vortex to evenly suspend cells, 0.1 aliquots were removed from the tubes and spread on YEPD agar plates. 7. The plates were incubated at 37 degrees Celsius for 24 hours. 8. The resulting colonies were counted. Each colony is assumed to have risen from one cell

Data SDF Regular Tab Smart 5 min 185 119 208 104 174 115 155 134 143 75 116 97 137 84 159 86 125 91 95 45 min 101 176 102 126 129 163 135 114 175 128 138 117

Column 1 control SDF Column 2 Regular Column 3 Tap Column 4 Smart

Column 1 control SDF Column 2 Regular Column 3 Tap Column 4 Smart

5 min dunnett test T crit of 3.29 Tap Smart Water Regular Not Significant -1.275 .232 1.129

45 minute dunnett test T crit of 3.29 Regular Smart water Tap Significant Not significant Not Significant 3.97417 .001 .0001

Conclusion For both of the anova, 5 minutes and 45 minutes, the f value is greater then the f crit this means that the null can be rejected. The p value is below 0.05 this support that the null hypothesis was rejected. Dunnett shows all have no significant variation. This fails to reject the null hypothesis.The regular water has a significant variation to the control which reject the null.

Limitations Plating was slightly unsynchronized Only one concentration Only two exposure times Only one method was used.

Extension Add different variables Try different types of soda. Different test times See if a long exposure cause harm. Use different models

Works cited How Do Fertilizers Affect the Environment.” Environment News South Africa, Environment News South Africa, 18 Aug. 2018, www.environment.co.za/environmental- issues/how-do-fertilizers-affect-the-environment.html. “Why It's Time to Stop Punishing Our Soils with Fertilizers.” Yale E360, e360.yale.edu/features/why-its-time-to-stop-punishing-our-soils-with-fertilizers-and- chemicals. “How Fertilizers Harm Earth More Than Help Your Lawn.” Scientific American, www.scientificamerican.com/article/how-fertilizers-harm-earth/. Hunt, Janet. “Harmful Effects of Chemical Fertilizers.” Hunker.com, Hunker, 24 Oct. 2010, www.hunker.com/12401292/harmful-effects-of-chemical-fertilizers. Disabled World. “What Are Body Electrolytes and How Do They Work.” Disabled World, Disabled World, 22 Aug. 2013, www.disabled- world.com/health/electrolytes.php