References
Introduction
Hypothesis
Future Directions
Even though Vidatox has growth promoting characteristics in the experiments performed, using much lower concentrations could yield more promising results. Repeating all trials would be beneficial to ensure reliable and repeatable data. The limitations of testing and analyzing vidatox in wells rather than a living organism needs to be taken into consideration. Vidatox’s ability to increase the activity of the immune system will not be represented when testing in wells like it would in in vivo testing.
A long-term experiment could be testing the individual components of Vidatox that make it active. Vidatox may not be the most beneficial against S. Typhimurium, but a component of it might be. Once the active components that decrease the activity of S. Typhimurium are identified, comparing those components to compounds that chemically look similar might reveal possible S. Typhimurium inhibitors or antibiotics.
Department of Molecular, Cellular, and Developmental Biology
University of Colorado Boulder
Skylar Bates, Anneliese Eisele, Anna Fleming, Tiger Sutton
The effect of Vidatox on Salmonella Typhimurium growth
Abstract
University. Retrieved March 6, 2023, from
https://www.mcgill.ca/oss/article/health-quackery/blue-scorpion-venom-cancer
from https://www.ntnu.no/ub/scorpion-files/r_junceus.php
diabetic foot and immunology. Retrieved March 6, 2023, from https://www.cubamedic.net/en/cuban-medicines/vidatox/
NCBI. Retrieved April 3, 2023, from https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5403050/
hepatocellular carcinoma. Scientific Reports. Retrieved April 3, 2023, from https://www.nature.com/articles/srep44685
Results
Acknowledgments
Methods
Methods
Conclusions
Over the course of the semester, 3 different kinds of experiments were run to determine the effectiveness of Vidatox as an antibiotic. The bactericidal/bacteriostatic experiment was not performed because Vidatox did not show significant results like hits. From the data that we collected, there is no evidence to suggest that Vidatox is an antibiotic because it did not inhibit the growth of S. Typhimurium. At the max dose that can be administered to a human, (10 uM), Vidatox showed characteristics of being a growth promoter as it yielded high absorbance values on the spectrophotometer and even produced hits. Between the other doses, there was low variability in absorbance, with no statistical hits with the exception of the outlier in figure 4. This is considered an outlier as it does not fit with the trend of the graph and deviates from all other data.
Bacteria are capable of mutating rapidly and can share DNA between each other increasing
the problem of resistant
bacteria infections since these
evolutionary mechanisms allow
strains of bacteria to become
deadly due to growing multi-
drug resistance bacteria
(Figure 1.). Vidatox, extracted
from the poison of blue
scorpion (Rhopalurus Junceus),
is a potential antibiotic because it
is a readily available chemotherapy. Chemotherapies like Vidatox target cells that divide rapidly and have the potential to target other cells that divide rapidly which could include various types of bacteria like our model organism, S. Typhimurium. A very beneficial attribute of S. Typhimurium is that the bacteria induces typhoid fever-like symptoms in mice while only causing food poisoning in humans. Using S. Typhimurium is therefore safer to use within the lab while also being able to observe the effects of typhoid fever symptoms in mice. Thus, removing the risk of using a dangerous bacteria of S. Typhi which causes typhoid fever in humans.
Utilizing chemotherapies as antibiotics has been a popular approach in more recent years because similarly to antibiotics there is minimal disruption to the host, but significant damage to the pathogens (ScienceDirect 2023). Even though we are using a venom as our compound, the venom from the Blue Scorpion is harmless to people (McGill University, 2023). Vidatox has been proven to block vital nutrients from entering diseased cells which would be a beneficial function when fighting bacteria. Without the necessary nutrients, S. Typhimurium will not be able to survive. Vidatox has been proven to boost and strengthen the patient's immune system when digesting Vidatox (Cuba Medic 2022). This reaction results in an increased number of white blood cells being produced within the body which is vital for fighting off foreign pathogens or more specifically, tumors (Giovannini, 2017). Improving the function of the immune system could be a contributing factor in fighting off S. Typhimurium because when antibiotics are taken, the immune system still plays a huge role in curing the patient.
After the Golden Age of discovery, most of the easy to find antibiotics had been found and first generation techniques began to fail, (Rice & Galbraith, 2008) leaving no new classes of antibiotics to be found. While there is always a high demand for new antibiotics, pressure to maximize sales and profitability did not align with the appropriate use, promotion, or consumption of antibiotics. The overuse and misuse of antibiotics in the medical industry as well as the agriculture industry poses a great threat to the rapidly growing resistance of bacteria (Healey, 2022). In fact, bacterial infections are predicted to be the leading cause of death surpassing cancer by the year 2050 due to this lack of support. (Figure 1.) It is imperative that we allocate more resources towards finding new antibiotics for our future.
We would first like to acknowledge the University of Colorado Boulder and the Howard Hughes Medical Institute for funding this research lab and our experiments. Thank you to the MCDB Chair, Lee Niswander and the department for providing this opportunity. Additionally, we would like to thank the Principal Investigator, Corrie Detweiler for sponsoring this lab. Finally, thank you to Pamela Harvey, Vinny Scarato and our TA’s Corbin Miller, Geneva Todd and Aubrey Woodard for their support throughout the semester.
Figure 1. Deaths From Drug-Resistant Infections Set To Skyrocket
Figure 2. Max Dose of Vidatox (10uM) compared to DMSO and Ampicillin. Vidatox showed some growth promoting hits because some results were +2 standard deviations away from the mean. During this experiment, the average of all max dose trials yielded no evidence to suggest a hit. The mean was still much higher than the negative control, showing growth promoting characteristics.
Figure 4. Dose response curve for a 1:10 dilution. There is no clear trend in the graph. All but the 0.01µM concentration were not hits which was likely caused by an error during pipetting. The .01 is an outlier as it does not fit with the rest of the data.
Figure 3. Dose response curve for a 1:2 dilution. The graph shows that there is a noticeable trend that dips down at a concentration of 1.25 uM. The absorbance value then increases after 1.25 uM showing that in this experiment, the lowest concentration of vidatox had the largest absorbance value. However, none of the concentrations presented hits.
Max Dose
1:2 Dilution Protocol
of Vidatox starting at 5µM.
50µg/ml Ampicillin in 6 wells containing 90µL of S. Typhimurium
1:10 Dilution Protocol
Hits are two standard deviations higher or two standard deviations lower than the class mean of DMSO compiled from compound screening. Two standard deviations higher an effective growth promoter. Two standard deviations lower indicates an effective antibiotic. Any data that lies in between ± two standard deviations is a non-hit which is insufficient to draw a conclusion about the attributes of the solution.
**All plates were incubated at 37°C for a 24 hour period before analytical examination. Absorbance data was collected from a spectrophotometer read at 620 nm for each individual plate. All S. Typhimurium is sourced from Su Lab CU Boulder cultured in M9 Minimal Media.
If Chemotherapies like Vidatox can target other cells that divide rapidly like tumors, then Vidatox will be able to kill or stop the growth of rapidly dividing bacteria which will significantly lower the concentration of bacteria in tested wells.