Impact of Sample Preservation and Manipulation on Insect Gut Microbiome Profiling. A Test Case With Fruit Flies (Diptera, Tephritidae)

Bibliographic Details
Title: Impact of Sample Preservation and Manipulation on Insect Gut Microbiome Profiling. A Test Case With Fruit Flies (Diptera, Tephritidae)
Authors: Maarten De Cock, Massimiliano Virgilio, Peter Vandamme, Antonios Augustinos, Kostas Bourtzis, Anne Willems, Marc De Meyer
Source: Frontiers in Microbiology, Vol 10 (2019)
Publisher Information: Frontiers Media S.A., 2019.
Publication Year: 2019
Collection: LCC:Microbiology
Subject Terms: amplicon sequencing, gut microbiome, methodology, Tephritidae, Ceratitis capitata, Microbiology, QR1-502
More Details: High-throughput sequencing (HTS) techniques are of great value for the investigation of microbial communities, and have been extensively used to study the gut microbiome. While most studies focus on the human gut, many others have investigated insects. However, because of the rapid spread of HTS techniques, a lot of variation exists in the protocols for sample preparation. In the present study, we investigated the impact of two widely adopted sample-processing procedures preceding library preparation, i.e., preservation of insect tissue in 70% ethanol (EtOH) and sample dissection. We used the fruit fly Ceratitis capitata (Diptera: Tephritidae) as a model organism and set up two experiments, one comparing the effects of sample manipulation and preservation across life stages and the other across fruit samples from different sources. The results of this study showed no major effects of dissection on the outcome of HTS. However, EtOH preservation did have effects on the recovered gut microbiome, the main effect being a significant reduction of the dominant genus, Providencia, in EtOH-preserved samples. Less abundant bacterial groups were also affected resulting in altered microbial profiles obtained from samples preserved in 70% EtOH. These results have important implications for the planning of future studies and when comparing studies that used different sample preparation protocols.
Document Type: article
File Description: electronic resource
Language: English
ISSN: 1664-302X
Relation: https://www.frontiersin.org/article/10.3389/fmicb.2019.02833/full; https://doaj.org/toc/1664-302X
DOI: 10.3389/fmicb.2019.02833
Access URL: https://doaj.org/article/a9f081d68ea94a1eb0f73f992c56bb88
Accession Number: edsdoj.9f081d68ea94a1eb0f73f992c56bb88
Database: Directory of Open Access Journals
More Details
ISSN:1664302X
DOI:10.3389/fmicb.2019.02833
Published in:Frontiers in Microbiology
Language:English