Confirmation of biodegradation of low-density polyethylene in dark- versus yellow- mealworms (larvae of Tenebrio obscurus versus Tenebrio molitor) via. gut microbe-independent depolymerization

Tenebrio obscurus (Coleoptera: Tenebrionidae) larvae are capable of biodegrading polystyrene (PS) but their capacity for polyethylene (PE) degradation and pattern of depolymerization remains unknown. This study fed the larvae of T. obscurus and Tenebrio molitor, which have PE degrading capacity, two commercial low-density PE (LDPE) foams i.e., PE-1 and PE-2, with respective number-average molecular weights (M_n) of 28.9 and 27.3 kDa and weight-average molecular weights (M_w) of 342.0 and 264.1 kDa, over a 36-day period at ambient temperature. The M_w of residual PE in frass (excrement) of T. obscurus, fed with PE-1 and PE-2, decreased by 45.4 ± 0.4% and 34.8 ± 0.3%, respectively, while the respective decrease in frass of T. molitor was 43.3 ± 0.5% and 31.7 ± 0.5%. Data analysis showed that low molecular weight PE (<5.0 kDa) was rapidly digested while longer chain portions (>10.0 kDa) were broken down or cleaved, indicating a broad depolymerization pattern. Mass balance analysis indicated nearly 40% of ingested LDPE was digested to CO₂. Antibiotic suppression of gut microbes in T. molitor and T. obscurus larvae with gentamicin obviously reduced their gut microbes on day 15 but did not stop depolymerization because the M_n, Mw and size- average molecular weight (M_z) decreased. This confirmed that LDPE biodegradation in T. obscurus was independent of gut microbes as observed during previous PS degradation in T. molitor, suggesting that the intestinal digestive system could perform LDPE depolymerization. High-throughput sequencing revealed significant shifts in the gut microbial community during bran-fed and unfed conditions in response to LDPE feeding in both Tenebrio species. The respective predominant gut genera of Spiroplasma sp. and Enterococcus sp. were observed in LDPE-fed T. molitor and T. obscurus larvae.