Anthropogenic and biological activities elevate microplastics pollution in headwater ecosystem of Yangtze tributaries in Hindu Kush-Himalayan region.

Microplastic (MP) pollution is widely spread in oceans, freshwater, and terrestrial environments but MPs in mountainous headwater ecosystem are rarely reported. This study focuses on the headwater of Yangtze tributaries of the Hindu Kush-Himalayan (HKH) region. Five streams at elevations of 900 to 3300 m were selected to investigate the distribution of MPs in water and sediments across altitudes. MPs were found in all water and sediment samples from top stream zone nearly in absence of anthropogenic activity, low anthropogenic zone, and high anthropogenic zone, increased from 12-54, 81-185 to 334-847 items/L, and 2-35, 26-84 to 124-428 items/kg, respectively. This elevation-dependent MP distribution indicated that as elevation decreased, anthropogenic activities intensified and increased MPs input and their abundance, size, and diversity. Notably, hydraulic projects, such as damming, were identified as potential barriers to the migration of MPs downstream. Microbiome analyses revealed the presence of bacterial genes associated with plastic biodegradation in all sediment samples. The study indicates that Shangri-la mountainous streams have been polluted with MPs for years with potential risk of generation of nano-sized particles via natural fragmentation and biodegradation, and thus raises concern on MPs pollution in headwaters streams in mountainous regions.

Biodegradation of various grades of polyethylene microplastics by Tenebrio molitor and Tenebrio obscurus larvae: Effects on their physiology.

Polyethylene (PE) is the most productive plastic product and includes three major polymers including high-density polyethylene (HDPE), linear low-density polyethylene (LLDPE) and low-density polyethylene (LDPE) variation in the PE depends on the branching of the polymer chain and its crystallinity. Tenebrio obscurus and Tenebrio molitor larvae biodegrade PE. We subsequently tested larval physiology, gut microbiome, oxidative stress, and PE degradation capability and degradation products under high-purity HDPE, LLDPE, and LDPE powders (<300 µm) diets for 21 days at 65 ± 5% humidity and 25 ± 0.5 °C. Our results demonstrated the specific PE consumption rates by T. molitor was 8.04-8.73 mg PE ∙ 100 larvae-1⋅day-1 and by T. obscurus was 7.68-9.31 for LDPE, LLDPE and HDPE, respectively. The larvae digested nearly 40% of the ingested three PE and showed similar survival rates and weight changes but their fat content decreased by 30-50% over 21-day period. All the PE-fed groups exhibited adverse effects, such as increased benzoquinone concentrations, intestinal tissue damage and elevated oxidative stress indicators, compared with bran-fed control. In the current study, the digestive tract or gut microbiome exhibited a high level of adaptability to PE exposure, altering the width of the gut microbial ecological niche and community diversity, revealing notable correlations between Tenebrio species and the physical and chemical properties (PCPs) of PE-MPs, with the gut microbiome and molecular weight change due to biodegradation. An ecotoxicological simulation by T.E.S.T. confirmed that PE degradation products were little ecotoxic to Daphnia magna and Rattus norvegicus providing important novel insights for future investigations into the environmentally-friendly approach of insect-mediated biodegradation of persistent plastics.

Molecular-Weight-Dependent Degradation of Plastics: Deciphering Host-Microbiome Synergy Biodegradation of High-Purity Polypropylene Microplastics by Mealworms.

The biodegradation of polypropylene (PP), a highly persistent nonhydrolyzable polymer, by Tenebrio molitor has been confirmed using commercial PP microplastics (MPs) (Mn 26.59 and Mw 187.12 kDa). This confirmation was based on the reduction of the PP mass, change in molecular weight (MW), and a positive Δδ13C in the residual PP. A MW-dependent biodegradation mechanism was investigated using five high-purity PP MPs, classified into low (0.83 and 6.20 kDa), medium (50.40 and 108.0 kDa), and high (575.0 kDa) MW categories to access the impact of MW on the depolymerization pattern and associated gene expression of gut bacteria and the larval host. The larvae can depolymerize/biodegrade PP polymers with high MW although the consumption rate and weight losses increased, and survival rates declined with increasing PP MW. This pattern is similar to observations with polystyrene (PS) and polyethylene (PE), i.e., both Mn and Mw decreased after being fed low MW PP, while Mn and/or Mw increased after high MW PP was fed. The gut microbiota exhibited specific bacteria associations, such as Kluyvera sp. and Pediococcus sp. for high MW PP degradation, Acinetobacter sp. for medium MW PP, and Bacillus sp. alongside three other bacteria for low MW PP metabolism. In the host transcriptome, digestive enzymes and plastic degradation-related bacterial enzymes were up-regulated after feeding on PP depending on different MWs. The T. molitor host exhibited both defensive function and degradation capability during the biodegradation of plastics, with high MW PP showing a relatively negative impact on the larvae.

Microplastic generation from field-collected plastic gauze: Unveiling the aging processes.

Accumulation of plastic debris in the environment is a matter of global concern. As plastic ages, it generates microplastic (MP) particles with high mobility. Understanding how MPs are generated is crucial to controlling this emerging contaminant. In this study, we utilized high-density polyethylene (HDPE) plastic gauze, collected from urban settings, as a representative example of plastic waste. The plastic gauze was subjected to various aging conditions, including freeze-thaw cycling, mechanical abrasion, and UV irradiation. Following aging, the plastic gauze was rinsed with water, and the number of generated MPs were quantified. It was found that aged plastic gauze generated up to 334 million MP particles per m2 (> 10 µm) during rinsing, a number two orders of magnitude higher than unaged plastic. Fragmentation occurred in two dimensions for bulk MPs of all morphotypes. However, specific aging approaches (i.e., mechanical abrasion and UV irradiation) generated spheres and fibers via pseudo-3D fragmentation. Additionally, changes in molecular weight, size distribution, and surface oxidation characteristics unveiled a complex pattern (i.e., irregular changes with exposure time). This complexity underscores the intricate nature of plastic debris aging processes in the environment.

Biodegradation of polyethylene terephthalate by Tenebrio molitor: Insights for polymer chain size, gut metabolome and host genes.

Polyethylene terephthalate (PET or polyester) is a commonly used plastic and also contributes to the majority of plastic wastes. Mealworms (Tenebrio molitor larvae) are capable of biodegrading major plastic polymers but their degrading ability for PET has not been characterized based on polymer chain size molecular size, gut microbiome, metabolome and transcriptome. We verified biodegradation of commercial PET by T. molitor larvae in a previous report. Here, we reported that biodegradation of commercial PET (Mw 29.43 kDa) was further confirmed by using the δ13C signature as an indication of bioreaction, which was increased from - 27.50‰ to - 26.05‰. Under antibiotic suppression of gut microbes, the PET was still depolymerized, indicating that the host digestive enzymes could degrade PET independently. Biodegradation of high purity PET with low, medium, and high molecular weights (MW), i.e., Mw values of 1.10, 27.10, and 63.50 kDa with crystallinity 53.66%, 33.43%, and 4.25%, respectively, showed a mass reduction of > 95%, 86%, and 74% via broad depolymerization. Microbiome analyses indicated that PET diets shifted gut microbiota to three distinct structures, depending on the low, medium, and high MW. Metagenome sequencing, transcriptomic, and metabolic analyses indicated symbiotic biodegradation of PET by the host and gut microbiota. After PET was fed, the host's genes encoding degradation enzymes were upregulated, including genes encoding oxidizing, hydrolyzing, and non-specific CYP450 enzymes. Gut bacterial genes for biodegrading intermediates and nitrogen fixation also upregulated. The multiple-functional metabolic pathways for PET biodegradation ensured rapid biodegradation resulting in a half-life of PET less than 4 h with less negative impact by PET MW and crystallinity.

Unveiling Fragmentation of Plastic Particles during Biodegradation of Polystyrene and Polyethylene Foams in Mealworms: Highly Sensitive Detection and Digestive Modeling Prediction.

It remains unknown whether plastic-biodegrading macroinvertebrates generate microplastics (MPs) and nanoplastics (NPs) during the biodegradation of plastics. In this study, we utilized highly sensitive particle analyzers and pyrolyzer-gas chromatography mass spectrometry (Py-GCMS) to investigate the possibility of generating MPs and NPs in frass during the biodegradation of polystyrene (PS) and low-density polyethylene (LDPE) foams by mealworms (Tenebrio molitor larvae). We also developed a digestive biofragmentation model to predict and unveil the fragmentation process of ingested plastics. The mealworms removed 77.3% of ingested PS and 71.1% of ingested PE over a 6-week test period. Biodegradation of both polymers was verified by the increase in the δ13C signature of residual plastics, changes in molecular weights, and the formation of new oxidative functional groups. MPs accumulated in the frass due to biofragmentation, with residual PS and PE exhibiting the maximum percentage by number at 2.75 and 7.27 µm, respectively. Nevertheless, NPs were not detected using a laser light scattering sizer with a detection limit of 10 nm and Py-GCMS analysis. The digestive biofragmentation model predicted that the ingested PS and PE were progressively size-reduced and rapidly biodegraded, indicating the shorter half-life the smaller plastic particles have. This study allayed concerns regarding the accumulation of NPs by plastic-degrading mealworms and provided critical insights into the factors controlling MP and NP generation during macroinvertebrate-mediated plastic biodegradation.

Biodegradation of polyvinyl chloride, polystyrene, and polylactic acid microplastics in Tenebrio molitor larvae: Physiological responses.

It is widely understood that microplastics (MPs) can induce various biological stresses in macroinvertebrates that are incapable of biodegrading plastics. However, the biodegradation and physiological responses of plastic-degrading macroinvertebrates toward MPs of different degradability levels remain unexplored. In this study, Tenebrio molitor larvae (mealworms) were selected as a model of plastics-degrading macroinvertebrate, and were tested against three common plastics of different degradability rankings: polyvinyl chloride (PVC), polystyrene (PS), and polylactic acid (PLA) MPs (size <300 µm). These three MPs were biodegraded with the rate sequence of PLA > PS > PVC, resulting in a reversed order of negative physiological responses (body weight loss, decreased survival, and biomass depletion) of mealworms. Simultaneously, the levels of reactive oxygen species (ROS), antioxidant enzyme activities, and lipid peroxidation were uniformly increased as polymer degradability decreased and intermediate toxicity increased. PVC MPs exhibited higher toxicity than the other two polymers. The oxidative stresses were effectively alleviated by supplementing co-diet bran. The T. molitor larvae fed with PLA plus bran showed sustainable growth without an increase in oxidative stress. The results provide new insights into the biotoxicity of MPs on macroinvertebrates and offer comprehensive information on the physiological stress responses of plastic-degrading macroinvertebrates during the biodegradation of plastics with different degradability levels.

Responses of gut microbiomes to commercial polyester polymer biodegradation in Tenebrio molitor Larvae.

Polyethylene terephthalate (PET) is a mass-produced fossil-based plastic polymer that contributes to catastrophic levels of plastic pollution. Here we demonstrated that Tenebrio molitor (mealworms) was capable of rapidly biodegrading two commercial PET resins (microplastics) with respective weight-average molecular weight (Mw) of 39.33 and 29.43 kDa and crystallinity of 22.8 ± 3.06% and 18 ± 2.25%, resulting in an average mass reduction of 71.03% and 73.28% after passage of their digestive tract, and respective decrease by 9.22% and 11.36% in Mw of residual PET polymer in egested frass. Sequencing of 16 S rRNA gene amplicons of gut microbial communities showed that dominant bacterial genera were enriched and associated with PET degradation. Also, PICRUSt prediction exhibited that oxidases (monooxygenases and dioxygenases), hydrolases (cutinase, carboxylesterase and chitinase), and PET metabolic enzymes, and chemotaxis related functions were up-regulated in the PET-fed larvae. Additionally, metabolite analyses revealed that PET uptake caused alterations of stress response and plastic degradation related pathways, and lipid metabolism pathways in the T. molitor larvae could be reprogrammed when the larvae fed on PET. This study provides new insights into gut microbial community adaptation to PET diet under nutritional stress (especially nitrogen deficiency) and its contribution to PET degradation.

Gut Microbiome Associating with Carbon and Nitrogen Metabolism during Biodegradation of Polyethene in Tenebrio larvae with Crop Residues as Co-Diets.

Tenebrio molitor and Tenebrio obscurus (Coleoptera: Tenebrionidae) larvae are two commercial insects that eat plant and crop residues as diets and also biodegrade synthetic plastics polyethylene (PE). We examined biodegradation of low-density PE (LDPE) foam (Mn = 28.9 kDa and Mw = 342.0 kDa) with and without respective co-diets, i.e., wheat brain (WB) or corn flour (CF), corn straw (CS), and rice straw (RS) at 4:1 (w/w), and their gut microbiome and genetic metabolic functional groups at 27.0 ± 0.5 °C after 28 days of incubation. The presence of co-diets enhanced LDPE consumption in both larvae and broad-depolymerized the ingested LDPE. The diet type shaped gut microbial diversity, potential pathways, and metabolic functions. The sequence of effectiveness of co-diets was WB or CF > CS > RS for larval development and LDPE degradation. Co-occurrence networks indicated that the larvae co-fed with LDPE displayed more complex correlations of gut microbiome than the larvae fed with single diets. The primary diet of WB or CF and crop residues CS and RS provided energy and nitrogen source to significantly enhance LDPE biodegradation with synergistic activities of the gut microbiota. For the larvae fed LDPE and LDPE plus co-diets, nitrogen fixation function was stimulated compared to normal diets and associated with LDPE biodegradation.

Influence of Polymer Size on Polystyrene Biodegradation in Mealworms (Tenebrio molitor): Responses of Depolymerization Pattern, Gut Microbiome, and Metabolome to Polymers with Low to Ultrahigh Molecular Weight.

Biodegradation of polystyrene (PS) in mealworms (Tenebrio molitor lavae) has been identified with commercial PS foams. However, there is currently limited understanding of the influence of molecular weight (MW) on insect-mediated plastic biodegradation and the corresponding responses of mealworms. In this study, we provided the results of PS biodegradation, gut microbiome, and metabolome by feeding mealworms with high-purity PS microplastics with a wide variety of MW. Over 24 days, mealworms (50 individuals) fed with 0.20 g of PS showed decreasing removal of 74.1 ± 1.7, 64.1 ± 1.6, 64.4 ± 4.0, 73.5 ± 0.9, 60.6 ± 2.6, and 39.7 ± 4.3% for PS polymers with respective weight-average molecular weights (Mw) of 6.70, 29.17, 88.63, 192.9, 612.2, and 1346 kDa. The mealworms degraded most PS polymers via broad depolymerization but ultrahigh-MW PS via limited-extent depolymerization. The gut microbiome was strongly associated with biodegradation, but that with low- and medium-MW PS was significantly distinct from that with ultrahigh-MW PS. Metabolomic analysis indicated that PS biodegradation reprogrammed the metabolome and caused intestinal dysbiosis depending on MW. Our findings demonstrate that mealworms alter their gut microbiome and intestinal metabolic pathways in response to in vivo biodegradation of PS polymers of various MWs.

Microplastics in urban runoff: Global occurrence and fate.

Public concerns on microplastic (MP) pollution and its prevalence in urban runoff have grown exponentially. Huge amounts of MPs are transported from urban environments via surface runoff to different environment compartments, including rivers, lakes, reservoirs, estuaries, and oceans. The global concentrations of MPs in urban runoff range from 0 to 8580 particles/L. Understanding the sources, abundance, composition and characteristics of MPs in urban runoff on a global scale is a critical challenge because of the existence of multiple sources and spatiotemporal heterogeneity. Additionally, dynamic processes in the mobilization, aging, fragmentation, transport, and retention of MPs in urban runoff have been largely overlooked. Furthermore, the MP flux through urban runoff into rivers, lakes and even oceans is largely unknown, which is very important for better understanding the fate and transport of MPs in urban environments. Here, we provide a critical review of the global occurrence, transport, retention process, and sinks of MPs in urban runoff. Relevant policies, regulations and measures are put forward. Future global investigations and mitigation efforts will require us to address this issue cautiously, cooperating globally, nationally and regionally, and acting locally.

Nanoplastic stimulates metalloid leaching from historically contaminated soil via indirect displacement.

It is generally believed that the ability of nanoplastics (NPs) to mobilize other contaminants is due to direct adsorption; however, this intuitive belief is questioned in this study when it comes to a historically contaminated soil where mining activities since 1958 have resulted in arsenic (As) and cadmium (Cd) enrichment. Negatively charged polystyrene (PS) NPs were used in this study, which should theoretically stimulate Cd (metal cation) instead of As (negatively charged oxyanion) leaching if direct adsorption accounted for co-transport. Surprisingly, PS NPs enhanced the leaching of As by up to over 5 times (p value < 0.05), but had almost no effect on Cd leaching (p value > 0.05). A novel indirect displacement model was therefore developed to describe the phenomenon of enhanced As leaching. It has been found that negatively charged NPs interacted with As via competition for soil binding sites. Underlying mechanistic insights were further explored via both theoretical calculations with the Extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) approach, and direct characterization using Scanning Electron Microscopy (SEM) and Computed X-ray Microtomography (µCT) showing binding sites and flow channels, respectively. The overall results provide new and valuable insights into NP-metal(loid) interactions in the natural soil environment, which can be integrated in future studies regarding the transport and risk assessment of NPs, and toxic metal(loid)s.

Impacts of physical-chemical property of polyethylene on depolymerization and biodegradation in yellow and dark mealworms with high purity microplastics.

Yellow and dark mealworms (Tenebrio molitor and Tenebrio obscurus) biodegrade commercial polyethylene (PE) materials at a high rate. We examined the impact of physical and chemical properties on biodegradation using high purity microplastics (MPs). These included high-density polyethylene (HDPE), low-density polyethylene (LDPE), and linear low-density polyethylene (LLDPE), all with different weight average molecular weights (Mw) and different crystallinity degrees in T. molitor and T. obscurus larvae. The biodegradation extent in the two mealworms was similar but strongly depended on the polymer type in sequence, since LDPE > LLDPE> HDPE (with respective Mw of 222.5, 110.5 and 182 kDa). When LDPE MPs with Mw of 0.84, 6.4 and 106.8 kDa and HDPE with Mw of 52, 105 and 132.7 kDa were tested, the PE MPs with lower Mw showed a greater extent of depolymerization. The results of dominance analysis indicated that less branching structure and higher crystallinity degree negatively impacted depolymerization and biodegradation. Py-GC/MS analysis confirmed the breaking of the macromolecule backbone as well as the formation of oxidized functional groups after all the tested PE materials passed through the mealworm intestine. The results demonstrated that molecular weight, PE type, branching, and crystallinity degree significantly affect the biodegradation capability of PE by the mealworms, and possibly by other biological systems as well.

Response of the yellow mealworm (Tenebrio molitor) gut microbiome to diet shifts during polystyrene and polyethylene biodegradation.

Plastic biodegradation by mealworm is regarded as an emerging strategy for plastic disposal. In this study, the polystyrene (PS) and low density polyethylene (LDPE) degradation efficiency by yellow mealworms (Tenebrio molitor larvae) supplemented with bran and the effects of plastics on the gut core microbiome were explored to construct a circular and continuous reactor for plastic biodegradation in the future. The gut microbiome was also investigated with dietary shift to explore the relationship between specific diets and gut microbes. The bran plus plastic (7:1 ratio, w/w) co-diet contributed to the mealworm survival and growth. The formation of -CË­O-/-C-O- groups in the plastic-fed mealworms frass represented the oxidation process of plastic biodegradation in the mealworm gut. The changes in molecular weights (Mw, Mn and Mz) of residual PS and LDPE in mealworms frass compared with that of PS and PE feedstock confirmed the plastic depolymerization and biodegradation. Lactobacillus and Mucispirillum were significantly associated with PE + bran diet compared to bran diet and PE diet, representing the response of mealworm gut microbiome to the bran and plastic mixture was distinguished from either bran or plastics alone. The gut microbiome changed substantially with the diet shift, indicating that microbial community assembly was a stochastic process and diverse plastic-degrading bacteria might occur in the mealworm gut.

Vertical migration of microplastics in porous media: Multiple controlling factors under wet-dry cycling.

Terrestrial soils are not only a large reservoir for Microplastics (MPs), but also a possible entrance to the subsurface environment, posing potential risks to the subterranean habitats and groundwater. In this study, we examined the vertical transport of MPs of four polymers, i.e., polyethylene (PE), polyethylene terephthalate (PET), polypropylene (PP) and polyamide (PA), in porous sand media driven by wet-dry cycling. The effects of polymer properties, MP size, sand particle size, wet-dry cycles, and dissolved organic matter (DOM) on their migration behavior were investigated. Surface hydrophobicity showed a strong positive correlation with MP mobility, with PA exhibiting the greatest movement potential, followed by PE, PET, and PP. The penetration depth of MP particles increased with decreasing MP particle size (dMP) and increasing sand diameter (dsand). MP particles migrated deeper in sand media when dMP/dsand < 0.11. Furthermore, frequent wet-dry cycles and the presence of DOM promoted the vertical migration of MPs in the sand. The results revealed multiple factors influencing the vertical migration of MPs in sand, which is instructive for understanding the ecological risk of MPs in potentially contaminated soil (e.g., farmland with long-term mulching) to the subsurface environment and potential negative impact to public health.

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.

Modeling the Conditional Fragmentation-Induced Microplastic Distribution.

Microplastics (MPs) are drawing increasing attention from the international community due to their potential threats to the ecosystem and human health. Although their occurrence and spatial distribution have been extensively studied in recent years, the relationship between their abundance and sizes remains unclear. Moreover, the underlying mechanisms dominating their size distribution have rarely been explored. In the present study, we developed a novel conditional fragmentation model to describe MP size distribution in the soil environment. It is proposed that the distribution of MPs is not a coincidence but controlled by conditional aging. The applicability of this model was tested using data collected from different land use settings in Beijing, China. A distinct downsizing phenomenon from fibers, films, and fragments to granules is observed. Undisturbed land use types accumulated larger sized MPs with higher stability, while human interference accelerated the fragmentation of MPs. Both morphological analysis and time-of-flight secondary ion mass spectroscopy (TOF-SIMS) observations provided direct evidence for the conditional fragmentation process. Furthermore, the model has proven to be suitable for describing the size distribution of MPs from various sources (including atmospheric deposition, transportation, and agriculture) and aging processes (such as mechanical abrasion, chemical oxidation, and photochemical transformation). It is proposed that this model can be used for various purposes in MP-related studies, especially source identification, transport modeling, and risk assessment.

Biodegradation of polylactic acid by yellow mealworms (larvae of Tenebrio molitor) via resource recovery: A sustainable approach for waste management.

Polylactic acid (PLA) is biodegraded rapidly under composting or thermophilic temperature but slowly under natural conditions with substantial microplastics generated. In this study, we examined the feasibility of PLA biodegradation and developed a novel approach for PLA waste management using yellow mealworms (Tenebrio molitor larvae) to achieve biodegradation and resource recovery simultaneously. Results confirmed PLA biodegradation in mealworms as sole PLA and PLA-bran mixtures (10%, 20%, 30% and 50% PLA, wt/wt). Feeding PLA-bran mixtures supported the larval development with higher survival rates and lower cannibal rates than feeding PLA only at ambient temperature. The PLA conversion efficiency was 90.9% with 100% PLA diet and was around 81.5-86.9% with PLA-bran mixtures. A peak insect biomass yield was achieved at a PLA ratio of 20%. PLA biodegradation was verified via detection of chemical and thermal modifications. Gut microbial community analysis indicated that intestinal communities shifted with PLA biodegradation, resulting in clusters with OTUs unique to the PLA diet. Based on these findings, we propose a circular approach for PLA waste management via resource recovery of used PLA as the feedstock for insect biomass production, management of mealworm excrement waste as fertilizer, and utilization of agricultural products for PLA production.

Characterization of biodegradation of plastics in insect larvae.

Biodegradation of plastics has been observed at rapid turnover rate by some insect larvae, especially those of Coleoptera, in particular Tenebrionidae. Tenebrio molitor larva is well studied and capable of biodegrading polystyrene (PS), polyethylene (PE), polypropylene (PP), and polyvinyl chloride (PVC) in their digestive intestine in synergy with their gut microflora. This chapter includes the methods, protocols, and procedures used to characterize biodegradation of plastics in T. molitor larvae and their gut microbiomes with polystyrene as the model feedstock. The methods used can be expanded to enable investigation of other plastics and/or insects.

Enhanced Bioavailability and Microbial Biodegradation of Polystyrene in an Enrichment Derived from the Gut Microbiome of Tenebrio molitor (Mealworm Larvae).

As the global threat of plastic pollution has grown in scale and urgency, so have efforts to find sustainable and efficient solutions. Research conducted over the past few years has identified gut environments within insect larvae, including Tenebrio molitor (yellow mealworms), as microenvironments uniquely suited to rapid plastic biodegradation. However, there is currently limited understanding of how the insect host and its gut microbiome collaborate to create an environment conducive to plastic biodegradation. In this work, we provide evidence that T. molitor secretes one or more emulsifying factor(s) (30-100 kDa) that mediate plastic bioavailability. We also demonstrate that the insect gut microbiome secretes factor(s) (<30 kDa) that enhance respiration on polystyrene (PS). We apply these insights to culture PS-fed gut microbiome enrichments, with elevated rates of respiration and degradation compared to the unenriched gut microbiome. Within the enrichment, we identified eight unique gut microorganisms associated with PS biodegradation including Citrobacter freundii, Serratia marcescens, and Klebsiella aerogenes. Our results demonstrate that both the mealworm itself and its gut microbiome contribute to accelerated plastic biodegradation. This work provides new insights into insect-mediated mechanisms of plastic degradation and potential strategies for cultivation of plastic-degrading microorganisms in future investigations and scale-up.