Researcher(s)
- Amelia Owen, Chemical Engineering, University of Delaware
Faculty Mentor(s)
- Mark Blenner, Chemical and Biomolecular Engineering, University of Delaware
- Rachel Keown, Biological Sciences, University of Delaware
Abstract
Low-density polyethylene (LDPE) is a widely used plastic due to its beneficial properties and cost effectiveness, making it one of the most common types of plastic found in landfills. However, its ubiquity and resistance to biodegradation make it a growing environmental concern. Recent studies have shown that Tenebrio molitor (mealworms) can survive on an LDPE-only diet, and may contribute to plastic degradation through their microbial activity. Bacteriophages, viruses that infect bacteria, are present in the microbiome of mealworms but have not been well explored. This project aims to investigate how phages may influence the gut microbiome of mealworms fed LDPE, and how they affect plastic-degrading bacteria.
In this investigation, mealworms were fed diets of bran (no plastic control) or LDPE. Gut samples were collected at intervals over 30 days on either plastic or control diets. Following each sampling, centrifugation was performed to separate mealworm tissue and bacteria cells, isolating the viral fraction in the resulting supernatant. Any remaining host (18s) or bacterial (16s) genes were targeted using PCR and if present, removed through the use of DNase. Proteinase K was then applied to dissolve the viral protein capsid, releasing the viral DNA. Phenol-chloroform-isoamyl (PCI) extraction was performed to purify the viral DNA for downstream sequencing. Based on the number of mealworms sampled, we expect to obtain DNA at the scale of 10s to 100s of nanograms that should be suitable for long-read sequencing. This method offers a new approach to measure the viral DNA yield from the gut samples of mealworms, allowing us to further study virus-host interactions of mealworms on different diets. Improving this method will allow for more efficient sequencing in future research, which can provide more information into how bacteriophages interact with bacterial communities involved in plastic degradation.