The main objectives of this study were to investigate the applicability of chitosan as an effective wood preservative against subterranean termites, conduct metagenomic analysis of the bacterial hindgut community of Reticulitermes flavipes exposed to chitosan-treated wood, and perform chitosanase activity assay of metagenomics suggested bacterial species potentially responsible for chitosan breakdown. Chitosan showed termiticidal effects on subterranean termites at varying retention levels. Termite mortality increased when exposed to samples treated with higher chitosan concentration solutions. Approximately 40 - 100% of chitosan retained in treated-wood was leached depending on the initial retention. Post-leaching results indicate chitosan is not suitable for protection against both subterranean termites in outdoor conditions, but should be effective in non-leaching/indoor applications. For metagenomic analysis of the bacterial hindgut community of Reticulitermes flavipes, two methods were used for sequence data interpretation. The Illumina BaseSpace program identified twenty-six bacteria phyla with significant differences in abundance between the chitosan-treated and control groups. The second method, mothur, identified fifteen bacterial phyla also with significant differences in abundance between both treatment groups. Similar bacterial taxa were uniquely assigned to samples from termites fed on chitosan-treated wood using both methods. These results suggest a treatment driven effect on the hindgut bacteria diversity. While majority of the bacterial taxa were common to both methods, inconsistencies detected using the BaseSpace program suggests that the Greengenes database in its present state is not reliable for 16S rRNA gene sequence analysis. As for chitosanase activity of bacterial species with significance abundance from chitosan-treated wood exposed termites, three bacteria species, Lactococcus raffinolactis, Lactococcus lactis, and Dysgonomonas gadei, were examined. After culturing on chitosan media plates and broth, no conclusive activity could be detected from all three species. Further studies need to be conducted to understand the mechanism of chitosan toxicity to termites and insects in general and to prevent chitosan leaching from treated wood. A comparative metatransciptomic study needs to be implemented to supplement the metagenomic study performed herein, so as to elucidate the exact bacteria species involved in chitosan breakdown and the enzymes produced. Also, other bacterial species suggested by the metagenomic data to possess chitosanase activity should be investigated.
| Identifer | oai:union.ndltd.org:MSSTATE/oai:scholarsjunction.msstate.edu:td-2643 |
| Date | 08 December 2017 |
| Creators | Raji, Olanrewaju Adeyemi |
| Publisher | Scholars Junction |
| Source Sets | Mississippi State University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Theses and Dissertations |
Page generated in 0.0022 seconds