Published: July 21st, 2023
Nasonia wasp embryos were dissected from Lucillia sericata pupae after parasitization for 12-24 h and washed with alcohol and 10% sodium hypochlorite solution to obtain germ-free embryos. After rearing the germ-free embryos and supplying them with Nasonia rearing medium to grow and develop in vitro, germ-free Nasonia adults were obtained.
Aseptic rearing technology is a method of culturing insects under sterile or almost sterile conditions, which can effectively eliminate the influence of external microorganisms on insect microbiota and thus promote the rapid development of insect microbiota research. Nasonia (wasp genus) is a parasitic wasp insect that has many advantages, such as a short lifespan, high genetic variation, easy operation, etc., and is widely used as an insect model system. Unlike antibiotic treatment, which can only reduce the number of microorganisms in animals, aseptic rearing techniques can control both the composition and quantity of microorganisms in animals, further facilitating the study of host-microbe interactions. However, previous versions of Nasonia rearing medium (NRM) have some defects and problems, such as a complex and time-consuming preparation process, easy contamination by bacteria or fungi, and short storage time. Therefore, this study solves these problems by optimizing the tools used in the NRM preparation process, storage conditions, and component ratios. The optimized medium could allow storage at -20 °C for at least 3 months and eliminate the possibility of NRM contamination during feeding sterile wasps. This further improves the survival rate and health level of aseptic Nasonia, which is important for using Nasonia as a model for microbial research.
Germ-free animals are animals that have no detectable living microorganisms and parasites1. Germ-free embryos can be obtained by dissecting the mother under aseptic conditions and subsequently raised in barrier systems2. Such animals can be used to study the effects of microorganisms on animals, such as on the intestinal microbiota, immune system, and metabolism1. With certain technical means, many insects and even mammals can be rendered sterile3,4. Germ-free animals have a unique role and have been widely used in va....
1. Preparation of germ-free Nasonia rearing medium
The preparation efficiency of NRM was greatly improved by improving the preparation tools. In addition, the problem of NRM pollution in the feeding process was eliminated by optimizing the strategy and preservation method. At the same time, the adjusted NRM had a more suitable nutritional ratio for the growth and development of germ-free wasps with L. sericata as hosts. The survival rate of germ-free wasps from larvae to pupae was significantly improved compared to germ-free wasps reared with NRMv3 using GFRv2 (.......
With the application of high-throughput detection technologies such as genomics and metabolomics, researchers have gradually realized that there is huge genetic diversity and metabolic complexity in the gut microbiota16. These symbiotic bacteria are closely related to various physiological or pathological states, such as host nutritional metabolism, tumors, immunity, and aging through complex interactions with the host17. However, the research related to the network of the .......
Funding: this work was supported by the National Science Foundation of China (32270538), the National Key R&D Program of China (2022YFF0710603), the Natural Science Foundation of Beijing (6222046), and the CAS strategic funding via the CAS-CSIRO funding scheme (152111KYSB20210011) awarded to G.H.W. Author contributions: all authors developed the scope and focus of the review and contributed to the writing of the manuscript.....
|0.22 Sterile vacuum filter
|1x PBS solution
|200 mesh nylon net
|8, 1.2, 0.8, and 0.45 µm filters
|Shanghai Xingya Purification Material Factory
|Absolute ethyl alcohol
|Commercial Drosophila Medium
|No Catalog numbers
|Purchase on Taobao
|Lucillia sericata pupae
|Hefei Dayuan Biotechnology Co., Ltd.
|No Catalog numbers
|Purchase on Taobao
|Small writing brush
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