In this project, an outgrowth and optical density (OD) measurement step was developed and validated for an established protocol for antibiotic treatment of Anopheles stephensi mosquitoes. This step allows the elimination of uncleared mosquitoes from experimental pools. Our findings suggest that, as previously demonstrated, antibiotic treatment reduced the mosquito microbiome to an extent, as evidenced by decreased bacterial growth observed through OD readings of our experimental samples. Additionally, qPCR analysis of 16S abundance provided quantitative data on bacterial abundance, which further supported the effectiveness of antibiotic treatment. Finally, a comparison of mosquito samples pooled randomly or with high OD individuals removed showed a significant improvement in bacterial clearance in samples with our refinement step included. The method established here can generate experimental samples for third-generation sequencing with minimal bacterial contamination for robust hypothesis testing. The addition of OD measurement is a rapid, cost-effective step that can potentially improve hypothesis testing on large pools of samples. This study contributes to our understanding of the impact of antibiotic treatment on mosquito microbiomes. Improvements in molecular and bench-level techniques can greatly contribute to our ability to gain insights into the role of microbes in mosquito biology.
| Identifer | oai:union.ndltd.org:unt.edu/info:ark/67531/metadc2332653 |
| Date | 05 1900 |
| Creators | Qadri, Tanvi |
| Contributors | Macias, Vanessa Michelle, 1986-, Henard, Calvin, Azad, Rajeev K. |
| Publisher | University of North Texas |
| Source Sets | University of North Texas |
| Language | English |
| Detected Language | English |
| Type | Thesis or Dissertation |
| Format | Text |
| Rights | Public, Qadri, Tanvi, Copyright, Copyright is held by the author, unless otherwise noted. All rights Reserved. |
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