Non-caloric artificial sweeteners are among the most commonly used food additives in the world, consumed by both healthy and obese individuals. People like the "no calorie" or "no guilt" aspect that comes along with this zero calorie sweetener when choosing between regular and diet soda or any other sweetened treat. What people do not realize however is that these artificial sweeteners can actually lead to obesity indirectly by affecting the plethora of microbiota in the human gut and affecting glucose intolerance. While people are possibly choosing the no calorie option because they think it is healthier and want to lose weight, they do not realize that they could actually be causing more damage and weight gain than they would with just regular sugar. Diabetes and obesity is a huge problem, especially in America. About nine percent of the American population suffers from diabetes and about one third of the population is considered to be obese, so as much research as possible should be conducted in order to prevent this trend from growing.

It has been previously proven in both mice and humans that changes in the gut microbiota after consuming high, yet FDA acceptable, amounts of sweetener leads to a greater likelihood of developing diabetes. Instead of testing on living subjects, this research uses an in-vitro approach in testing the direct effects of a realistic amount of sweetener on the microbiota. Using a standardized in-vitro digestion method, the sweeteners are digested and then exposed to two types of bacteria found in the microbiome, Escherichia Coli and Staphylococcus Epidermidis. The growth curves of these bacteria cultures are plotted using absorption spectroscopy, and the different curves are compared to a control.

This experiment consists of ten experimental groups, five for each type of bacteria. The first group is a control group of bacteria growing in the digest medium, but no type of sweetener is present. The second group is the bacteria growing in a culture of digested sucrose, or normal sugar. The third, fourth, and fifth groups are bacteria cultures growing in saccharin, sucralose, and aspartame respectively. Each trial is done for each type of bacteria and repeated three times for a total of thirty experimental groups.

Since each trial was repeated three times, the values were averaged together and graphed into growth curves that were then compared. The results from each of the three trials were all close in number with relatively small standard deviations. For both types of bacteria, comparing the control graph and the

growth in sucrose showed very little statistical difference. For example, sucrose only inhibited growth of E.Coli by two percent. However, for both types of bacteria, the three growth curves from medium with artificial sweeteners showed significant differences from the control group. For E.Coli, aspartame increased growth by 14.5 percent, saccharin inhibited growth by 18 percent, and sucralose inhibited growth by 36 percent. For Staphy.Epi, aspartame increased growth by 33 percent, saccharin inhibited growth by 58 percent, and sucralose inhibited growth by 78 percent.