Establishment and Maintenance of Gnotobiotic American Cockroaches (Periplaneta americana).
Establishment and Maintenance of Gnotobiotic American Cockroaches (Periplaneta americana).
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DOI：
10.3791/61316
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发表时间：
2021-05-28
影响因子：
1.2
通讯作者：
Ottesen, Elizabeth A.
中科院分区：
文献类型：
--
作者：
Dukes, Helen E.;Dyer, Josey E.;Ottesen, Elizabeth A.
Gnotobiotic Animal是对微生物组结构和功能的控制的强大工具这里将昆虫定义为蟑螂，这些蟑螂仍然含有垂直传播的内共生体（blattabacterium），但缺乏通常驻留的其他微生物在该方案的表面和消化道上，卵子病例（卵子）被从（非遗产）菌落中取出，一旦收集并进行了灭菌。 6周的脑心脏输注（BHI）琼脂，直到它们孵化或因污染而被移除确保若虫不是在给定条件下无法在BHI上生长的微生物，额外的质量控制措施使用限制性片段长度多态性（RFLP）来测试使用这种方法的非雌激素若虫。接种简单或复杂的微生物群落，并用作肠道微生物组研究的工具。
该方案用于在孵化前对卵子病例（othecae）进行灭菌，以建立和维持gnotobiotic美国蟑螂（Periplaneta Americana）。
Gnotobiotic animals are a powerful tool for the study of controls on microbiome structure and function. Presented here is a protocol for the establishment and maintenance of gnotobiotic American cockroaches (Periplaneta americana). This approach includes built-in sterility checks for ongoing quality control. Gnotobiotic insects are defined here as cockroaches that still contain their vertically transmitted endosymbiont (Blattabacterium) but lack other microbes that normally reside on their surface and in their digestive tract. For this protocol, egg cases (oothecae) are removed from a (nonsterile) stock colony and surface sterilized. Once collected and sterilized, the oothecae are incubated at 30 °C for approximately 4–6 weeks on brain-heart infusion (BHI) agar until they hatch or are removed due to contamination. Hatched nymphs are transferred to an Erlenmeyer flask containing a BHI floor, sterile water, and sterile rat food. To ensure that the nymphs are not housing microbes that are unable to grow on BHI in the given conditions, an additional quality control measure uses restriction fragment-length polymorphism (RFLP) to test for non-endosymbiotic microbes. Gnotobiotic nymphs generated using this approach can be inoculated with simple or complex microbial communities and used as a tool in gut microbiome studies. This protocol is used in establishing and maintaining gnotobiotic American cockroaches (Periplaneta americana) by surface sterilizing the egg cases (oothecae) prior to hatching. These gnotobiotic insects contain their vertically transmitted Blattabacterium endosymbionts but have axenic guts.
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影响因子：
2.5
作者：
Nicklas, Werner;Keubler, Lydia;Bleich, Andre
通讯作者：
Bleich, Andre
影响因子：
17.1
作者：
Faith JJ;Ahern PP;Ridaura VK;Cheng J;Gordon JI
通讯作者：
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影响因子：
2.7
作者：
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Nishino H
DOI：
10.1073/pnas.0907504106
发表时间：
2009-11-17
影响因子：
11.1
作者：
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通讯作者：
Moran, Nancy A.
影响因子：
4.4
作者：
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通讯作者：
Brune, Andreas