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Impact of sample preservation and manipulation on insect gut microbiome profiling : a test case with fruit flies (Diptera, Tephritidae)

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Abstract
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.
Keywords
amplicon sequencing, gut microbiome, methodology, Tephritidae, Ceratitis capitata, FLY CERATITIS-CAPITATA, BACTERIAL COMMUNITIES, BACTROCERA-DORSALIS, INTESTINAL-TRACT, LIFE-CYCLE, MICROORGANISMS, POPULATIONS, DIVERSITY, HOST, IDENTIFICATION

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MLA
De Cock, Maarten, et al. “Impact of Sample Preservation and Manipulation on Insect Gut Microbiome Profiling : A Test Case with Fruit Flies (Diptera, Tephritidae).” FRONTIERS IN MICROBIOLOGY, vol. 10, 2019, doi:10.3389/fmicb.2019.02833.
APA
De Cock, M., Virgilio, M., Vandamme, P., Augustinos, A., Bourtzis, K., Willems, A., & De Meyer, M. (2019). Impact of sample preservation and manipulation on insect gut microbiome profiling : a test case with fruit flies (Diptera, Tephritidae). FRONTIERS IN MICROBIOLOGY, 10. https://doi.org/10.3389/fmicb.2019.02833
Chicago author-date
De Cock, Maarten, Massimiliano Virgilio, Peter Vandamme, Antonios Augustinos, Kostas Bourtzis, Anne Willems, and Marc De Meyer. 2019. “Impact of Sample Preservation and Manipulation on Insect Gut Microbiome Profiling : A Test Case with Fruit Flies (Diptera, Tephritidae).” FRONTIERS IN MICROBIOLOGY 10. https://doi.org/10.3389/fmicb.2019.02833.
Chicago author-date (all authors)
De Cock, Maarten, Massimiliano Virgilio, Peter Vandamme, Antonios Augustinos, Kostas Bourtzis, Anne Willems, and Marc De Meyer. 2019. “Impact of Sample Preservation and Manipulation on Insect Gut Microbiome Profiling : A Test Case with Fruit Flies (Diptera, Tephritidae).” FRONTIERS IN MICROBIOLOGY 10. doi:10.3389/fmicb.2019.02833.
Vancouver
1.
De Cock M, Virgilio M, Vandamme P, Augustinos A, Bourtzis K, Willems A, et al. Impact of sample preservation and manipulation on insect gut microbiome profiling : a test case with fruit flies (Diptera, Tephritidae). FRONTIERS IN MICROBIOLOGY. 2019;10.
IEEE
[1]
M. De Cock et al., “Impact of sample preservation and manipulation on insect gut microbiome profiling : a test case with fruit flies (Diptera, Tephritidae),” FRONTIERS IN MICROBIOLOGY, vol. 10, 2019.
@article{8643088,
  abstract     = {{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.}},
  articleno    = {{2833}},
  author       = {{De Cock, Maarten and Virgilio, Massimiliano and Vandamme, Peter and Augustinos, Antonios and Bourtzis, Kostas and Willems, Anne and De Meyer, Marc}},
  issn         = {{1664-302X}},
  journal      = {{FRONTIERS IN MICROBIOLOGY}},
  keywords     = {{amplicon sequencing,gut microbiome,methodology,Tephritidae,Ceratitis capitata,FLY CERATITIS-CAPITATA,BACTERIAL COMMUNITIES,BACTROCERA-DORSALIS,INTESTINAL-TRACT,LIFE-CYCLE,MICROORGANISMS,POPULATIONS,DIVERSITY,HOST,IDENTIFICATION}},
  language     = {{eng}},
  pages        = {{13}},
  title        = {{Impact of sample preservation and manipulation on insect gut microbiome profiling : a test case with fruit flies (Diptera, Tephritidae)}},
  url          = {{http://doi.org/10.3389/fmicb.2019.02833}},
  volume       = {{10}},
  year         = {{2019}},
}

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