Biodegradation of Polyvinyl Chloride (PVC) in Tenebrio molitor (Coleoptera: Tenebrionidae) larvae.

Tenebrio molitor larvae (Coleoptera: Tenebrionidae) are capable of depolymerizing and biodegrading polystyrene and polyethylene. We tested for biodegradation of Polyvinyl Chloride (PVC) in T. molitor larvae using rigid PVC microplastic powders (MPs) (70-150 μm) with weight-, number-, and size-average molecular weights (Mw, Mn and Mz) of 143,800, 82,200 and 244,900 Da, respectively, as sole diet at 25 °C. The ingestion rate was 36.62 ± 6.79 mg MPs 100 larvae-1 d-1 during a 16-day period. The egested frass contained about 34.6% of residual PVC polymer, and chlorinated organic carbons. Gel permeation chromatography (GPC) analysis indicated a decrease in the Mw, Mn and Mz by 33.4%, 32.8%, and 36.4%, respectively, demonstrating broad depolymerization. Biodegradation and oxidation of the PVC MPs was supported by the formation of OC and OC functional groups using frontier transform infrared spectroscopy (FTIR) and 1H nuclear magnetic resonance (1H NMR), and by significant changes in the thermal characteristics using thermo-gravimetric analysis (TGA). Chloride released was counted as about 2.9% of the PVC ingested, indicating limited mineralization of the PVC MPs. T. molitor larvae survived with PVC as sole diet at up to 80% over 5 weeks but did not complete their life cycle with a low survival rate of 39% in three months. With PVC plus co-diet wheat bran (1:5, w/w), they completed growth and pupation as same as bran only in 91 days. Suppression of gut microbes with the antibiotic gentamicin severely inhibited PVC depolymerization, indicating that the PVC depolymerization/biodegradation was gut microbe-dependent. Significant population shifts and clustering in the gut microbiome and unique OTUs were observed after PVC MPs consumption. The results indicated that T. molitor larvae are capable of performing broad depolymerization/biodegradation but limited mineralization of PVC MPs.

Peng BY ，Chen Z ，Chen J ，Yu H ，Zhou X ，Criddle CS ，Wu WM ，Zhang Y ... -  《-》

Biodegradation of expanded polystyrene and low-density polyethylene foams in larvae of Tenebrio molitor Linnaeus (Coleoptera: Tenebrionidae): Broad versus limited extent depolymerization and microbe-dependence versus independence.

Yellow mealworms (Tenebrio molitor larvae) are capable of biodegrading polystyrene (PS) and low-density polyethylene (LDPE). This study tested biodegradation of one expanded PS (EPS) with a weight-average molecular weight (Mw) 256.4 kDa and two LDPE foams with respective Mw of 130.6 kDa (PE-1) and 288.7 kDa (PE-2) in T. monitor larvae obtained in Beijing, China. The larvae consumed EPS and both LDPEs over a 60 day. Fourier transform infrared spectroscopy and thermogravimetric analyses of frass confirmed the formation of new oxygen-containing functional groups, as well as a change in physical property and chemical modification, indicating that biodegradation of EPS and LDPE occurred. Gel permeation chromatography analysis confirmed broad depolymerization of EPS and PE-1 (i.e., a decrease in both Mw and a number-average molecular weight (Mn)) but revealed limited extent depolymerization of PE-2 (i.e., increase in Mn and decrease in Mw). For all materials, the size-average molecular weight (Mz) was decreased. Biodegradation and oxidation of EPS and LDPE were confirmed using FTIR and TGA analysis. Depression of gut microbes by the antibiotic gentamicin resulted in significant inhibition of EPS depolymerization but did not stop LDPE depolymerization, resulting in the increase in Mn and revealing that PS biodegradation was gut microbe-dependent but LDPE biodegradation was less dependent or independent of gut microbes. Gut microbial community analysis indicated that, as expected, under different dietary conditions, the intestinal flora significantly shifted to communities associated with biodegradation of EPS and LDPE. The results indicated the complexity and limitation of biodegradation of plastics in plastics-eating T. molitor larvae.

Yang L ，Gao J ，Liu Y ，Zhuang G ，Peng X ，Wu WM ，Zhuang X ... -  《-》

Biodegradation of polypropylene by yellow mealworms (Tenebrio molitor) and superworms (Zophobas atratus) via gut-microbe-dependent depolymerization.

Polypropylene (PP), a fossil-based polyolefin plastics widely used worldwide, is non-hydrolyzable and resistant to biodegradation as a major source of plastic pollutants in environment. This study focused on feasibility of PP biodegradation in the larvae of two species of darkling beetles (Coleoptera: Tenebrionidae) i.e., yellow mealworms (Tenebrio molitor) and superworms (Zophobas atratus) using PP foam with number-, weight-, and size-average molecular weights (Mn, Mw, and Mz) of 109.8, 356.2, and 765.0 kDa, respectively. The tests were conducted in duplicates with respective larvae (300 T. molitor and 200 Z. atratus each incubator) at 25 °C and 65% humidity for over a 35-day period. The larvae of T. molitor and Z. atratus fed with PP foam as sole diet consumed PP at 1.0 ± 0.4 and 3.1 ± 0.4 mg 100 larvae-1 days-1, respectively; when fed the PP foam plus wheat bran, the consumption rates were enhanced by 68.11% and 39.70%, respectively. Gel permeation chromatography analyses of the frass of T. molitor and Z. atratus larvae fed PP only indicated that Mw was decreased by 20.4 ± 0.8% and 9.0 ± 0.4%; Mn was increased by 12.1 ± 0.4% and 61.5 ± 2.5%; Mz was decreased by 33.8 ± 1.5% and 32.0 ± 1.1%, indicating limited extent depolymerization. Oxidation and biodegradation of PP was confirmed through analysis of the residual PP in frass. Depression of gut microbes with the antibiotic gentamicin inhibited PP depolymerization in both T. molitor and Z. atratus larvae. High throughput 16S rRNA sequencing revealed that Citrobacter sp. and Enterobacter sp. were associated with PP diets in the gut microbiome of Z. atratus larvae while Kluyvera was predominant in the T. molitor larvae. The results indicated that PP can be biodegraded in both T. molitor and Z. atratus larvae via gut microbe-dependent depolymerization with diversified microbiomes.

Yang SS ，Ding MQ ，He L ，Zhang CH ，Li QX ，Xing DF ，Cao GL ，Zhao L ，Ding J ，Ren NQ ，Wu WM ... -  《-》

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 (Mn) of 28.9 and 27.3 kDa and weight-average molecular weights (Mw) of 342.0 and 264.1 kDa, over a 36-day period at ambient temperature. The Mw 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 Mn, Mw and size- average molecular weight (Mz) 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.

Yang SS ，Ding MQ ，Zhang ZR ，Ding J ，Bai SW ，Cao GL ，Zhao L ，Pang JW ，Xing DF ，Ren NQ ，Wu WM ... -  《-》

Biodegradation of low-density polyethylene and polystyrene in superworms, larvae of Zophobas atratus (Coleoptera: Tenebrionidae): Broad and limited extent depolymerization.

Larvae of Zophobas atratus (synonym as Z. morio, or Z. rugipes Kirsch, Coleoptera: Tenebrionidae) are capable of eating foams of expanded polystyrene (EPS) and low-density polyethylene (LDPE), similar to larvae of Tenebrio molitor. We evaluated biodegradation of EPS and LDPE in the larvae from Guangzhou, China (strain G) and Marion, Illinois, U.S. (strain M) at 25 °C. Within 33 days, strain G larvae ingested respective LDPE and PS foams as their sole diet with respective consumption rates of 58.7 ± 1.8 mg and 61.5 ± 1.6 mg 100 larvae-1d-1. Meanwhile, strain M required co-diet (bran or cabbage) with respective consumption rates of 57.1 ± 2.5 mg and 30.3 ± 7.7 mg 100 larvae-1 d-1. Fourier transform infrared spectroscopy, proton nuclear magnetic resonance, and thermal gravimetric analyses indicated oxidation and biodegradation of LDPE and EPS in the two strains. Gel permeation chromatography analysis revealed that strain G performed broad depolymerization of EPS, i.e., both weight-average molecular weight (Mw) and number-average molecular weight (Mn) of residual polymers decreased, while strain M performed limited extent depolymerization, i.e., Mw and Mn increased. However, both strains performed limited extent depolymerization of LDPE. After feeding antibiotic gentamicin, gut microbes were suppressed, and Mw and Mn of residual LDPE and EPS in frass were basically unchanged, implying a dependence on gut microbes for depolymerization/biodegradation. Our discoveries indicate that gut microbe-dependent LDPE and EPS biodegradation is present within Z. atratus in Tenebrionidae, but that the limited extent depolymerization pattern resulted in undigested polymers with high molecular weights in egested frass.

Peng BY ，Li Y ，Fan R ，Chen Z ，Chen J ，Brandon AM ，Criddle CS ，Zhang Y ，Wu WM ... -  《-》