Abstract
The massive production of plastics, together with inadequate waste management, has intensified the problem of plastic pollution. Recently, the scientific community has increasingly investigated the interactions between plastic polymers and biological systems, with particular interest in plastivorous insects and their gut-associated microbial communities. The lesser mealworm Alphitobius diaperinus has emerged as a promising model for studying biological interactions associated with polystyrene (PS) modification during gut transit. In this study, two complementary approaches were applied to characterize gut mediated responses to PS. First, chemical analyses were employed to provide evidence of polymer modification following gut transit; micro-FTIR analysis confirmed the presence of PS residues in larval frass and showed detectable structural changes in the recovered material, while GC-MS analysis identified α-methylstyrene and cumyl alcohol as compounds associated with PS chemical modifications and not detected in controls. Second, gut microbiota composition was assessed through metabarcoding analysis using full-length PacBio HiFi sequencing of the 16S rDNA gene. Differences were observed between the microbiota of PS fed larvae and that of the control group under different experimental conditions. Together, these results indicate that PS ingestion leads to measurable chemical modifications of the polymer and shifts in gut microbial composition, providing insight into insect–microbe interactions associated with PS transformation.
Data availability
Raw data were deposited in NCBI’s SRA database within BioProject ID PRJNA1330245, SRA numbers: SRS26581475, SRS26581478, SRS26581479, SRS26581482, SRS26581483, SRS26581486, SRS26581487, SRS26581489 - SRS26581491, SRS26581495, SRS26581497, SRS26581498, SRS26581502, SRS26581503, SRS26581506, SRS26581509, SRS26581510, SRS26581512, SRS26581515, SRS26581517 - SRS26581520, SRS26581522, SRS26581526, SRS26581528, SRS26581530, SRS26581531, SRS26581535, SRS26581536, SRS26581538, SRS26581539, SRS26581541, SRS26581544, SRS26581548, SRS26581549, SRS26581551, SRS26581552, SRS26581554, SRS26581555, SRS26581558, SRS26581560, SRS26581562 - SRS26581564, SRS26581566, SRS26581571 - SRS26581573, and Bio-Sample numbers: SAMN51481962 - SAMN51482011. All supporting data, code and protocols have been provided within the article, through supplementary data files or on the following link: https://github.com/ESZlab/Alphitobius_metabarcoding.
References
Bucci, K., Tulio, M. & Rochman, C. M. What is known and unknown about the effects of plastic pollution: a meta-analysis and systematic review. Ecol. Appl. 30(2), e02044 (2020).
Plastics Europe. Plastics – the Fast Facts 2025. Plastics Europe. https://plasticseurope.org/knowledge-hub/plastics-the-fast-facts-2025/. Accessed 24 Feb 2026 (2026).
Lamba, P., Kaur, D. P., Raj, S. & Sorout, J. Recycling/reuse of plastic waste as construction material for sustainable development: a review. Environ. Sci. Pollut. Res. 29(57), 86156–86179 (2022).
Li, P. et al. Characteristics of plastic pollution in the environment: a review. Bull. Environ. Contam. Toxicol. 107, 577–584 (2021).
Gallo, F. et al. Marine litter plastics and microplastics and their toxic chemicals components: the need for urgent preventive measures. Environ. Sci. Eur. 30, 1–14 (2018).
Germanov, E. S., Marshall, A. D., Bejder, L., Fossi, M. C. & Loneragan, N. R. Microplastics: no small problem for filter-feeding megafauna. Trends Ecol. Evol. 33(4), 227–232 (2018).
Xu, L. Progress in polystyrene biodegradation by insect gut microbiota. World J. Microbiol. Biotechnol. 40(5), 143 (2024).
Boctor, J., Pandey, G., Xu, W., Murphy, D. V. & Hoyle, F. C. Nature’s plastic predators: a comprehensive and bibliometric review of plastivore insects. Polymers 16(12), 1671 (2024).
Dar, M. A. et al. Current paradigms and future challenges in harnessing gut bacterial symbionts of insects for biodegradation of plastic wastes. Insect Sci. 32(3), 726–752 (2025).
Asiandu, A. P., Wahyudi, A. & Sari, S. W. A review: Plastics waste biodegradation using plastics-degrading bacteria. J. Environ. Treat. Tech. 9(1), 148–157 (2021).
Ho, B. T., Roberts, T. K. & Lucas, S. An overview on biodegradation of polystyrene and modified polystyrene: the microbial approach. Crit. Rev. Biotechnol. 38(2), 308–320 (2018).
Hou, L. & Majumder, E. L. W. Potential for and distribution of enzymatic biodegradation of polystyrene by environmental microorganisms. Materials 14(3), 503 (2021).
Yuan, J., Cao, J., Yu, F. & Ma, J. Microbial degradation of polystyrene microplastics by a novel isolated bacterium in aquatic ecosystem. Sustain. Chem. Pharm. 30, 100873 (2022).
Yang, Y. et al. Biodegradation and mineralization of polystyrene by plastic-eating mealworms: part 2. role of gut microorganisms. Environ. Sci. Technol. 49(20), 12087–12093 (2015).
Peng, B. Y. et al. Biodegradation of polystyrene by dark (Tenebrio obscurus) and yellow (Tenebrio molitor) mealworms (Coleoptera: Tenebrionidae). Environ. Sci. Technol. 53(9), 5256–5265 (2019).
Yang, Y., Wang, J. & Xia, M. Biodegradation and mineralization of polystyrene by plastic-eating superworms Zophobas atratus. Sci. Total Environ. 708, 135233 (2020).
Kim, H. R. et al. Biodegradation of polystyrene by Pseudomonas sp. isolated from the gut of superworms (larvae of Zophobas atratus). Environ. Sci. Technol. 54(11), 6987–6996 (2020).
Wang, Z., Xin, X., Shi, X. & Zhang, Y. A polystyrene-degrading Acinetobacter bacterium isolated from the larvae of Tribolium castaneum. Sci. Total Environ. 726, 138564 (2020).
Woo, S., Song, I. & Cha, H. J. Fast and facile biodegradation of polystyrene by the gut microbial flora of Plesiophthalmus davidis larvae. Appl. Environ. Microbiol. 86(18), e01361-20 (2020).
Dunford, J. C., & Kaufman, P. E. Lesser mealworm, litter beetle, Alphitobius diaperinus (Panzer)(Insecta: Coleoptera: Tenebrionidae): EENY-367/IN662, rev. 6/2006. EDIS, (2006).
Francisco, O. & Do Prado, A. P. Characterization of the larval stages of Alphitobius diaperinus (Panzer)(Coleoptera: Tenebrionidae) using head capsule width. Rev. Bras. Biol. 61, 125–131 (2001).
Cucini, C. et al. Bacterial and fungal diversity in the gut of polystyrene-fed Alphitobius diaperinus (Insecta: Coleoptera). Animal Gene 17, 200109 (2020).
Cucini, C. et al. Polystyrene shaping effect on the enriched bacterial community from the plastic-eating Alphitobius diaperinus (Insecta: Coleoptera). Symbiosis 86(3), 305–313 (2022).
Zarra, F. et al. Novel insights into insect mediated polystyrene biodegradation through bacterial genome analyses. Sci. Rep. 15(1), 1047 (2025).
Tsochatzis, E., Lopes, J. A., Gika, H. & Theodoridis, G. Polystyrene biodegradation by Tenebrio molitor larvae: identification of generated substances using a GC-MS untargeted screening method. Polymers 13(1), 17 (2020).
Richard, C. M., Renault, D. & Colinet, H. Can the plastic-eating insect Alphitobius diaperinus be used as a new biodegradation agent for synthetic plastic insulator wastes?. Environ. Technol. Innov. 39, 104315 (2025).
Garrigós, M. C., Marín, M. L., Cantó, A. & Sánchez, A. Determination of residual styrene monomer in polystyrene granules by gas chromatography–mass spectrometry. J. Chromatogr. A 1061, 211–216 (2004).
Urbanek, A. K., Rybak, J., Hanus-Lorenz, B., Komisarczyk, D. A. & Mirończuk, A. M. Zophobas morio versus Tenebrio molitor: Diversity in gut microbiota of larvae fed with polymers. Sci. Total Environ. 952, 176005 (2024).
Ding, M. Q. et al. Effects of plastic aging on biodegradation of polystyrene by Tenebrio molitor larvae: insights into gut microbiome and bacterial metabolism. Sci. Total Environ. 953, 176130 (2024).
Tay, J. H., Asib, N., Aziz, N. A. A. & Tan, G. H. Biodegradation of expanded and extruded polystyrene with different diets by using Zophobas atratus larvae (Coleoptera: Tenebrionidae). Pertanika J. Trop. Agric. Sci. https://doi.org/10.47836/pjtas.46.2.06 (2023).
Ndotono, E. W., Tanga, C. M., Kelemu, S. & Khamis, F. M. Mitogenomic profiling and gut microbial analysis of the newly identified polystyrene-consuming lesser mealworm in Kenya. Sci. Rep. 14(1), 21370 (2024).
Yang, S. S. et al. Biodegradation of polystyrene wastes in yellow mealworms (larvae of Tenebrio molitor Linnaeus): factors affecting biodegradation rates and the ability of polystyrene-fed larvae to complete their life cycle. Chemosphere 191, 979–989 (2018).
Tsangaris, C. et al. Interlaboratory comparison of microplastic extraction methods from marine biota tissues: a harmonization exercise of the Plastic Busters MPAs project. Mar. Pollut. Bull. 164, 111992 (2021).
Wang, Y. & Zhang, Y. Investigation of gut-associated bacteria in Tenebrio molitor (Coleoptera: Tenebrionidae) larvae using culture-dependent and DGGE methods. Ann. Entomol. Soc. Am. 108(5), 941–949 (2015).
Posit team. Rstudio: Integrated Development Environment for R. Posit Software, PBC, Boston, MA. URL http://www.posit.co/ (2024).
Callahan, B. J. et al. DADA2: High-resolution sample inference from Illumina amplicon data. Nat. methods 13(7), 581–583 (2016).
Quast, C. et al. The SILVA ribosomal RNA gene database project: improved data processing and web-based tools. Nucleic Acids Res. 41(D1), D590–D596 (2012).
Wickham, H. ggplot2: Elegant Graphics for Data Analysis (Springer-Verlag, 2016).
McMurdie, P. J. & Holmes, S. Phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS One 8(4), e61217 (2013).
Martinez Arbizu, P. pairwiseAdonis: pairwise multilevel comparison using adonis. R package. Version 0.4. Available on URL: http://github.com/pmartinezarbizu/pairwiseAdonis. (2020)
Love, M., Anders, S. & Huber, W. Differential analysis of count data–The DESeq2 package. Genome Biol. 15(550), 10–1186 (2014).
Chen, H. & Boutros, P. C. VennDiagram: A package for the generation of highly-customizable Venn and Euler diagrams in R. BMC Bioinformatics 12(1), 35 (2011).
Acknowledgements
This work was supported by the NBFC to University of Siena/Department of Life Sciences, funded by the Italian Ministry of University and Research, PNRR, Missione 4 Componente 2, “Dalla ricerca all’impresa”, Investimento 1.4, Project CN00000033.
Author information
Authors and Affiliations
Contributions
Conceptualization and Software: F.Z. and C.C. Methodology and Validation: F.Z., R.F., C.C., C.P., M.B. and A.B. Formal analysis: F.Z. and F.N. Investigation and Data curation: F.Z. and R.F. Resources: C.P., A.B., F.N., A.C. and F.F. Writing – Original draft and Visualization: F.Z. Writing – Review & Editing: C.P., A.B., L.M., F.N., A.C. and F.F. Supervision: C.C., F.N. and F.F. Project administration: F.F. Funding acquisition: L.M., F.N., A.C. and F.F.
Corresponding authors
Ethics declarations
Competing interests
One of the authors (F.N.) is part of the Editorial Board of the journal but did not take part in the evaluation of this manuscript in any respect. Other authors declare no conflicts of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
Rights and permissions
Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
About this article
Cite this article
Zarra, F., Funari, R., Cucini, C. et al. Chemical characterization and gut microbial response unveil modification of polystyrene polymer in the lesser mealworm. Sci Rep (2026). https://doi.org/10.1038/s41598-026-44113-3
Received:
Accepted:
Published:
DOI: https://doi.org/10.1038/s41598-026-44113-3