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Effects of Vaping Liquid on Worm Behavior

Rebecca Cooke (rcooke@douglasps.net)

Faculty Mentor: Jagan Srinivasan (Department of Biology & Biotechnology)

Graduate Mentor: Samuel Isife

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C. elegans Preparation

Avoidance Assay

Chemotaxis Assay

References

Egg-Laying

Lesson Plan

Acknowledgements

Caenorhabditis elegans is a microscopic worm used as a model organism used to study the effects of vaping on behavior4

1. Birdsey J, Cornelius M, Jamal A, et al. Tobacco Product Use Among U.S. Middle and High School Students — National Youth Tobacco Survey, 2023. MMWR Morb Mortal Wkly Rep 2023; 72:1173–1182. DOI: http://dx.doi.org/10.15585/mmwr.mm7244a1

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2. Kalininskiy, A., Bach, C. T., Nacca, N. E., Ginsberg, G., Marraffa, J., Navarette, K. A., ... & Croft, D. P. (2019). E-cigarette, or vaping, product use associated lung injury (EVALI): case series and diagnostic approach. The Lancet Respiratory Medicine, 7(12), 1017-1026.

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3. National Center for Chronic Disease Prevention and Health Promotion (US) Office on Smoking and Health. E-Cigarette Use Among Youth and Young Adults: A Report of the Surgeon General [Internet]. Atlanta (GA): Centers for Disease Control and Prevention (US); 2016. Chapter 3, Health Effects of E-Cigarette Use Among U.S. Youth and Young Adults. Available from: https://www.ncbi.nlm.nih.gov/books/NBK538688/

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4. Henricson, A., Sonnhammer, E. L., Baillie, D. L., & Gomes, A. V. (2004). Functional characterization in Caenorhabditis elegans of transmembrane worm-human orthologs. BMC genomics, 5, 1-18.

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E-cigarettes (vapes) are currently the most popular nicotine delivery system among adolescents1

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  1. Worms were washed with 1 mL of M9 and distributed onto fresh NGM plates
  2. Added one drop of diH2O or 2M glycerol to the tail-end of a forward-moving worm and recorded behaviors.
  1. Worms were bleached to isolate embryos and synchronize ages of the worms.
  2. One L3 was placed on the Petri dish with OP50 and the vaping liquid dilutions.
  3. Number of eggs laid were counted each day for 5 days.

Vaping has been linked to:

  • Lung injury2 & Respiratory disease
  • Cardiovascular disease
  • Deficits in cognition, attention, and mood
  • Potential for long-term addiction3

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The goal of the research is to determine if exposure to vaping liquid changes worm behavior

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Wild-type worms were raised in 1%, 5%, and 10% vaping liquid stock mixed with OP50 E. coli bacteria as their food source

Vaping liquid stock created by diluting 20% with LB broth and autoclaved

This material is based upon work supported by the National Science Foundation under Grant No. EEC-2055507

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Thank you to Dr. Jagan Srinivasan, Samuel Isife, Caroline Muirhead, Elizabeth DiLoreto, Racheal Asaolu, and Aiyina Meng for their support and feedback.

Students will conduct similar experiments on C. elegans behavior to create a PSA on energy drinks and caffeine addiction.

Conclusion 1: Changes to worm attraction and avoidance behavior is concentration-dependent with worms showing reduced sensitivity to known attractants and repellents

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Conclusion 2: Worms lay fewer eggs when exposed to vaping liquid.

“Strengthen the prevention and treatment of substance abuse, including narcotic drug abuse and harmful use of alcohol.”

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n=10, Error bars are SEM, One-way ANOVA compared 0% vs. treatment, ns-no significance, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

Scan to see avoidance behavior!

n=10, Error bars are SEM, One-way ANOVA compared 0% vs. treatment, ns-no significance, *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001

 

1. Worms were washed with 1 mL of chemotaxis buffer

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  1. Worms were placed at the center of the chemotaxis plate containing diH2O and diacetyl on opposite ends of the plate.
  2. Allow worms to move on the plate for one hour.