Wax Worms (Galleria mellonella) as Potential Bioremediators for Plastic Pollution Student Researcher: Alexandria Elliott Faculty Mentor: Danielle Garneau, Ph.D. Center for Earth and Environmental Science SUNY Plattsburgh, Plattsburgh, NY 12901

Plastic Pollution Life History Stages Results Discussion • 30 million tons of plastic Combination of holes in plastic and • Egg stage: average length (0.478mm) • waste is generated annually and width (0.394mm) and persists 3-10 nylon in frass suggests worms are in the USA (Coalition 2018). days (Kwadha et al. 2017)(Fig. 3). digesting plastic (Figs. 6,7). • Larval stage: max length (30mm), white  Of the plastic pilot trials which • 50% landfill, < 10% cream in color, possess 3 apical teeth, exhibited signs of feeding, two were HDPE (Fig. 6). recycled (PlasticsEurope, and persists 22-69 days. A Plastics The Facts 2013) • Pre-pupal/Pupal stage: length (12- • Bombelli et al. (2017) and Yang et al. 20mm) and persists 3-12 and 8-10 days, (2014) found wax worms were capable respectively. All extremities are glued to of PE consumption. • 10% of world’s plastic waste body with molting substance.  Common bond (CH -CH ) in PE is  2 2 ends up in ocean 70% • Moth stage: sexual dimorphism is same as that in beeswax (Bombelli et sinks 30% floats in Fig. 1. Plastic Use distinct. Moths max length (20mm) and Fig. 5. Change in worm weight as a function of plastic pilot trial. al. 2017). Fig. 9. PE degradation as currents (Gyres, Fig. 2). (Plastics Europe). persists on average 6-14 days (males) B • Greater negative change in worm weight (g)/day for all  FT-IR shows degradation of PE (i.e., evidenced by FT-IR PE and 23 days (females). pilot trials consuming plastic (HDPE 1 & 2, PP, EVAC) vs. increase in surface roughness, novel peaks at 2,921 and 2,852 cm-1, new peak at ~3,350 controls (Fig. 5) plastic consumption may act as an spectral peaks) occurring with • Larvae of wax moth is partly responsible cm-1 ethylene glycol environmental stressor. digestion and PE mass loss in (Bombelli et al. 2017). for honeybee (Apis mellifera and A. experiments with a bacterial cerana) decline. They burrow into A B homogenate smear on plastic unsealed hive cells and throughout the (Bombelli et al. 2017; Fig. 9). honeycomb (Kwadha et al. 2017). • Yang et al. (2017) isolated two Dickman (1933) first noted wax worm C digestion of beeswax. bacterial strains from the digestive tracts Fig. 3. Wax worms A) life stages, B) of wax worms feeding on PE (Fig.10). honeycomb infestation, and C) sexual (HDPE) dimorphism. (Kwadha et al. 2017) • Plastic type may have altered Methods phenology of metamorphosis, such that Fig. 10. Wax worms on Wax worms were acquired from PetSmart and placed in screen-covered PE film and bacterial only 1/12 moths emerged, completing Ball jars and worms, frass, and plastics were weighed until moths isolates (Yang et al. Fig. 6. A) Produce bag (HDPE(2)) with foraging holes (blue circles) and, metamorphosis. emerged (Fig. 4). B) FT-IR spectrum (HDPE). 2014)  Those feeding on EVAC emerged 1st. A B C • Evidence of wax worm feeding was present on 2 HDPE Fig. 2. Ocean gyres and garbage patches (Scripps Research Inst.). samples, polypropylene isotactic, and ethylene/vinyl Overall Implications acetate (Fig. 6). • Common plastics are water bottles (PET), single- • Higher rates of worm weight loss may be evidence of plastic as a stressor worms are an impractical long-term organism for use low-density plastic bags (LDPE), bottle caps A B (polypropylene), fishing nets/line (nylon), and bioremediation of plastic. styrofoam (Plastics Europe). Fig. 4. A) Evaluating plastic B) weighing worms C) pupae inspection. • Future testing should focus on determining if gut contents (e.g., • Plastics photo- and mechanically degrade but Pilot 1: bacteria) digest plastic bacteria as long-term, large-scale persist as microplastics. Conditions: 5 worms per jar under 2 light regiments (darkness, natural bioremediation candidate for plastic pollution. light). Plastics: water bottle (Polyethylene teraphthalate (PET)), soda bottle (PET colored), PVC tubing (Poly(Diallyl phthalate (PDAP))), bottle caps (nylon) • Consumers must begin to reduce, recycle, and refuse plastic Bioremediation (Ethylene/Vinyl Acetate(EVAC)), and projector sheet (Hydroxypropyl dependence. Organisms who degrade environmental pollutants methyl cellulose). Acknowledgements Pilot 2: Fig. 7. A) Wax worm frass, B) FT-IR spectrum of frass (nylon). (e.g., bacteria, fungus, worms) Conditions: 5 worms per jar under 2 light regimes (natural, UV light). Many thanks to Dr. Mark Lesser for his assistance in statistical analysis Plastics: grocery bag (polyethylene, high density(HDPE(1))), produce bag • FT-IR analysis of frass using Program R. Additional thanks to the Lake Champlain Research Wax Worms (Galleria mellonella) (HDPE(2)), Ramen package (polypropylene, isotactic), bottle caps samples resulted in a Institute (Luke Meyers) for use of scales, Ball jars, and insect consultation. (EVAC), green mesh (HDPE(3)), and chocolate mold (polystyrene(PS)). Thanks to Erin Ashline FT-IR support. Support for this research and A B C best match for nylon in Controls: 5 worms per jar, under all 3 light conditions (darkness, natural trace amounts (Fig. 7). conference travel comes from the Center for Earth and Environmental and UV light), with NO plastics. Science and College Auxiliary Services of SUNY Plattsburgh. Pilot 3: Literature Cited Conditions: 10 worms per ball jar, under most favorable light regimes • There was no change in Bombelli, P., Howe, C., & Bertocchini, F. (2017). Polyethylene bio-degradation by caterpillars of the wax moth (natural and UV light) worm weight in Galleria mellonella. Current Biology, 27(8): R292-R293. Coalition, P. P. (2018). The Facts. Retrieved from Plastic Pollution Coalition Plastics: grocery bag (HDPE(1)), Ramen package (PP), produce bag darkness. Worm weight http://www.plasticpollutioncoalition.org/the-movement (HDPE(2)), and bottle caps (EVAC). declined in both light Dickman, R. 1933. Journal of Insect Physiology: Beeswax in the nutrition of the wax moth, Galleria mellonella Pilot 4: best conditions, highest performing plastics (L.). J. Cell. Comp. Physiol. 3, 223–246. conditions, albeit the Kwadha, A. C., Ong’amo, G. O., Ndegwa, P.N., Raina, S. K., & Fombong, A. T. (2017). The Biology and Control Fig. 3. A) wax worms, B) wax worm moth and pupae, Conditions: 10 worms, under UV light conditions. of the Greater Wax Moth, Galleria mellonella. Insects. Jun; 8(2): 61. doi: 10.3390/insects8020061 Plastic: produce bag (HDPE(1)) trend was insignificant Yang, J., Yang, Y, Wu W-M,m Zhao, J., Jiang, L. 2014. Evidence of polyethylene biodegradation by bacterial Ball jar array. Fig. 8. Light conditions strains from the guts of plastic-eating waxworms. Environmental Science and Technology. 8 (23), pp 13776– C) Replication: 3 replicates, one control worm jar (no plastics). (Fig.8). 13784.