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. 2024 Dec 31;10(6):364-368.
doi: 10.52601/bpr.2024.240025.

A rapid and reproducible method for generating germ-free Drosophila melanogaster

An-Qi Li  1   2 , Sha-Sha Li  2 , Hui-Lin Li  2 , Lan-Lan Zhong  3 , Guo-Bao Tian  3 , Xin-Yuan Zhao  2 , Qiao-Ping Wang  2   4   5
Affiliations

A rapid and reproducible method for generating germ-free Drosophila melanogaster

An-Qi Li et al. Biophys Rep. .

Abstract

Microbial communities exert a profound influence on various facets of animal behavior and physiology, making the comprehension of their interactions with hosts or the environment essential. Drosophila melanogaster, a widely recognized model organism, has been pivotal in elucidating host-microbe interactions. Despite the existence of several protocols for generating germ-free (GF) Drosophila, their reproducibility has been constrained by the technical difficulty of maintaining airtight conditions in centrifuge tubes. In this study, we introduce a refined method for the production of GF Drosophila, complemented by a straightforward verification process to ascertain its efficacy. We propose an innovative strategy employing bio-reaction tubes equipped with a 0.22 μm filter membrane cap, which facilitates the rearing and maintenance of GF flies, thereby streamlining the procedure and enhancing the efficiency of model construction.

Keywords: Axenic; Drosophila; Germ-free; Microbiota; Protocol.

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Conflict of interest statement

An-Qi Li, Sha-Sha Li, Hui-Lin Li, Lan-Lan Zhong, Guo-Bao Tian, Xin-Yuan Zhao and Qiao-Ping Wang declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
Sterile food in bio-reaction tubes. A Front view. B Top view
Figure 2
Figure 2
Assembly of egg collection cages. A The lid of egg collection cages. B The side walls of the egg collection cages are formed by cutting off the bottom of a clear plastic bushing. C The grape juice medium plates with yeast paste. D The fully assembled egg collection cage
Figure 3
Figure 3
Washing kit for egg dechorionation. A Sterile double-distilled water. B Sterile brushes. C Egg-washing containers. D 100 μm cell sieves
Figure 4
Figure 4
Verification of germ-free fruit fly. A Results of GF fly homogenate coated plate. B Results of MRS coated plate
See this image and copyright information in PMC

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