1. The Effects of Reiki on Bacteria Survivorship Jordan Ciccone Central Catholic High School

2. Reiki Japanese spiritual healing practice meaning “universal life energy.” Training is done by a Reiki Master. Reiki supposedly works via the practitioner intentionally willing the energy toward target areas. Either gets rid of negative energy or provides positive healing energy (disputed). Supposedly works on all forms of life.

3. Reiki (continued) Most practitioners use Reiki to complement Western medicine. Some use Reiki as preventive medicine. Few peer-reviewed scientific studies; no foundation in science. Critics propose the placebo effect as the source of Reiki’s effectiveness.

4. Escherichia coli A prokaryotic, gram negative, unicellular, bacterial cell which can most frequently be found in the large intestine of endothermic animals. May be used as an indicator organism for fecal contamination and has been a motive behind some product recalls. Used in this experiment because it is exemplified as a model prokaryote for scientific study due to its extensive documentation.

5. Cellular Stresses Several variables may be responsible for stressing and killing cells. Temperature Toxins Viral infections Radiation Heat Oxidation

6. Rationale The purpose of this experiment is to determine the effectiveness of Reiki on peroxide-stressed bacterial cells. Any significant difference in survivorship curves will lend scientific support for Reiki’s effectiveness with little possibility of a placebo effect.

7. Hypothesis 1. Null Reiki will not significantly alter the survivorship of peroxide- stressed E. coli. 2. Alternative Reiki will significantly increase the survivorship of peroxide- stressed E. coli.

8. Materials 40 LB agar plates(1% tryptone, 5% yeast extract, 1% NaCl, 2 mL 1M NaOH, 1.5% agar) YEPD Media (1% yeast extract, 2% peptone, 2% glucose (dextrose)) LB media (1% tryptone, 5% yeast extract, 1% NaCl) Klett spectrophotometer Sterile pipette tips Micropipettes Vortex Incubator Sidearm flask Spreading platform, spreader bar, ethanol 20 mL Sterile capped test tubes with Sterile Dilution Fluid (SDF) (10 mM KH 2 PO 4, 10 mM K 2 HPO 4, 1 mM MgSO 4, 0.1 mM CaCl 2, 100 mM NaCl) Escherichia coli 0.22 micron syringe filters + 10 mL syringe Reiki practitioner Two 1 Liter Beakers Water Thermometer Hydrogen Peroxide

9. Procedures 1. Escherichia coli cultures were grown overnight in sterile LB media,. 2. Samples of the overnight cultures were added to fresh medias in a sterile sidearm flask. 3. The cultures were placed in incubators (37°C) until a density of 50 Klett spectrophotometer units was reached. This represents a cell density of approximately 10 8 cells/ml. 5. The microbial cultures were serially diluted in sterile dilution fluid to a concentration of approximately 10 3 cells/ml. 6. The Reiki practitioner was permitted to prepare the other room with the identical setup for the treatment of the cell cultures within test tubes by the use of gestures and vocal harmonics. 7. 0.1 ml. of the E. coli culture was then added to 4 test tubes, yielding a final volume of 10 ml. in each tube. This step was repeated with four other test tubes for the identical set up in the other room. 8. Dilutions of hydrogen peroxide were then created in order to create an exposure concentration kill curve. The concentrations were 10^-5, 10^-4, 10^-3, 10^-1, and 0 (control).

10. Procedures (continued) 8. The E.coli cells were then exposed to the various concentrations of hydrogen peroxide. 9. The Reiki practitioner was then instructed to treat the cells with Reiki in that room only, by which she used tuning forks, quartz and celenite crystals in addition to gestures and vocal mantras to administer treatment to the cells. 10. After 15 minutes, an independent source color coded the four(stressed, non- stressed, treated stressed, treated non-stressed) tube groups with blue, green, black or red so as to mask the results to the experimenter for a double blind experiment. 11. After vortexing the test tubes to evenly suspend cells, 0.1 ml. aliquots were removed from one tube and spread on 5 plates of labeled with the same color. This was repeated for the other 7 tubes. Two tubes were used to help safeguard against procedural errors. 12. The plates were incubated at 37°C for 24 hours. 13. The resulting colonies were counted by the experimenter. Each colony is assumed to have derived from one cell. The colors were then identified as their respective variable groups by the independent source.

11. Experimental Layout Plate Layout (LB plates)  Experimental Layout (Test tubes and heat source) 

12. Plate Count Hydrogen Peroxide Exposure 0 ExposureE-5 H2O2E-4 H2O2E-3 H2O2E-1 H2O2 REIKI463, 452, 445, 435, 478, 421, 484 431 450, 480, 471, 432, 501, 412, 453, 437 450, 443, 400, 415, 420, 460, 418, 409 360, 400, 390, 373, 430, 401, 363, 432 2, 5, 3, 1, 1, 4, 2, 4 NO REIKI456, 434, 412, 475, 419, 437, 476, 429 412, 470, 494, 473, 462, 341, 525, 381 450, 447, 460, 475, 449, 435, 470, 446 455, 521, 434, 380, 380, 444, 400, 430 4, 13, 6, 8, 7, 7, 3, 12

13. LD50:.05 ( 5 * 10^-2 ) Colonies Exposure

14. H2O2 Exposure 0. 461177 0.008766 0.690562

15. ANOVA Analyses Exposure: 10^110^310^410^5 Control P-Value:0. 033420.0739390.0087660.6905620.461177 Reiki to Non-Reiki Groups

16. Dunnett’s Test Reiki Group T-crit: 3.02 Group T – Value Interpretation 10^-5 Not Significant 10^-4 Not Significant 10^-3 SIGNIFICANT 10^-1 SIGNIFICANT 0.299 2.145 5.086 39.659

17. Conclusion Dunnett’s Tests show that the Reiki therapy on E.coli exposed to oxidation concentrations of 10^-3 and 10^-1 had a significant effect on the survivorship of E. coli, lending support for the alternative hypothesis. Dunnett’s Tests show that the hydrogen peroxide concentration of 10^-1 had a significant effect on the survivorship of E. coli

18. Improvements ● More replications ● More trials ● Account for more variables ● Simultaneous plate spreading ● Analyzing Reiki’s effects in normal settings

19. References ● Centralcatholichs.com/biology