A cryopreservation method to recover laboratory- and field-derived bacterial communities from mosquito larval habitats.
A cryopreservation method to recover laboratory- and field-derived bacterial communities from mosquito larval habitats.
复制标题
DOI：
10.1371/journal.pntd.0011234
复制
发表时间：
2023-04
影响因子：
3.8
通讯作者：
中科院分区：
文献类型：
--
作者：
蚊子在广泛的水生栖息地中生长，其中含有高度多样化和可变的细菌群落，这些细菌群落塑造了幼虫和成虫的特征，包括某些蚊种的成年雌性将致病生物体传播给人类的能力。然而，虽然大多数蚊子研究控制了宿主基因型和环境条件，但微生物群变异对蚊子表型结果的影响往往未被考虑。无法对蚊子-微生物群相互作用进行可重复的实验室内和实验室间研究，也极大地限制了我们确定控制蚊媒疾病的微生物目标的能力。在这里，我们开发了一种方法来分离和冷冻保存来自黄热病蚊子埃及伊蚊（登革热、寨卡病毒和基孔肯雅病毒的主要载体）的实验室和现场幼虫饲养环境的细菌群落。然后，我们验证了使用我们的方法来生成由标准化实验室和现场衍生的细菌群落定殖的实验微观世界。我们的结果总体表明，与从非冷冻保存的新鲜材料中分离直接比较时，冷冻保存对实验室和现场来源的细菌的回收影响最小。我们的结果还表明，与新鲜材料相比，使用冷冻保存的菌种产生的复制微观世界中细菌群落的再现性有所提高。复制微观世界中的群落进一步捕获了实验室和野外幼虫环境中存在的大部分细菌多样性，尽管在含有野外来源细菌的微观世界中，恢复的类群与未恢复的类群相比，相对丰富度要低得多。总而言之，这些结果为蚊子研究的标准化提供了关键的下一步，包括确定的微生物群落定殖的幼虫饲养环境。它们还为蚊子与微生物相互作用的长期研究以及识别和操纵有可能降低蚊子传播能力的类群奠定了基础。蚊子在多种细菌群落的存在下发育，这些细菌群落塑造了它们传播致病病原体的能力。然而，我们目前对蚊子与微生物群相互作用的理解主要基于对低多样性细菌群落定殖的个体的研究，这些细菌群落通常与实验室或野外的蚊子无关。在这项研究中，我们开发了一种在实验室和现场从蚊子幼虫饲养环境中分离和冷冻保存微生物群的方法。然后，我们证明了这种方法的成功使用来产生由标准化微生物群落定殖的实验微观世界。我们的结果对媒介生物学领域具有至关重要的意义，因为它们将直接（i）在没有微生物群变异的混杂影响的情况下促进对感兴趣的蚊子性状（例如病原体易感性）的研究，以及（ii）实现蚊子与微生物群相互作用的可重复的实验室内和实验室间研究，以确定疾病控制的微生物目标。我们的结果也引起了微生物生态学和宿主-微生物相互作用领域的研究人员的广泛兴趣，因为它们展示了如何在其他系统中利用通常用于研究哺乳动物微生物群组装和功能的工具。
Mosquitoes develop in a wide range of aquatic habitats containing highly diverse and variable bacterial communities that shape both larval and adult traits, including the capacity of adult females of some mosquito species to transmit disease-causing organisms to humans. However, while most mosquito studies control for host genotype and environmental conditions, the impact of microbiota variation on phenotypic outcomes of mosquitoes is often unaccounted for. The inability to conduct reproducible intra- and inter-laboratory studies of mosquito-microbiota interactions has also greatly limited our ability to identify microbial targets for mosquito-borne disease control. Here, we developed an approach to isolate and cryopreserve bacterial communities derived from lab and field-based larval rearing environments of the yellow fever mosquito Aedes aegypti–a primary vector of dengue, Zika, and chikungunya viruses. We then validated the use of our approach to generate experimental microcosms colonized by standardized lab- and field-derived bacterial communities. Our results overall reveal minimal effects of cryopreservation on the recovery of both lab- and field-derived bacteria when directly compared with isolation from non-cryopreserved fresh material. Our results also reveal improved reproducibility of bacterial communities in replicate microcosms generated using cryopreserved stocks over fresh material. Communities in replicate microcosms further captured the majority of total bacterial diversity present in both lab- and field-based larval environments, although the relative richness of recovered taxa as compared to non-recovered taxa was substantially lower in microcosms containing field-derived bacteria. Altogether, these results provide a critical next step toward the standardization of mosquito studies to include larval rearing environments colonized by defined microbial communities. They also lay the foundation for long-term studies of mosquito-microbe interactions and the identification and manipulation of taxa with potential to reduce mosquito vectorial capacity. Mosquitoes develop in the presence of diverse bacterial communities that shape their ability to transmit disease-causing pathogens. However, our current understanding of mosquito-microbiota interactions is largely based on studies of individuals colonized by low-diversity communities of bacteria that are not commonly associated with mosquitoes in the laboratory or field. In this study, we developed an approach to isolate and cryopreserve microbiota from mosquito larval rearing environments in the lab and field. We then demonstrated the successful use of this approach to produce experimental microcosms colonized by standardized microbial communities. Our results are of critical significance to the field of vector biology because they will directly (i) facilitate the study of mosquito traits of interest (e.g., pathogen susceptibility) in the absence of confounding effects of microbiota variation, and (ii) enable reproducible intra- and inter-laboratory studies of mosquito-microbiota interactions to identify microbial targets for disease control. Our results are also of broad interest to researchers in the fields of microbial ecology and host-microbe interactions because they demonstrate how tools commonly used to study microbiota assembly and function in mammals can be leveraged in other systems.
登录
查看更多内容
影响因子：
4.2
作者：
Duguma D;Hall MW;Rugman-Jones P;Stouthamer R;Terenius O;Neufeld JD;Walton WE
通讯作者：
Walton WE
影响因子：
15.5
作者：
Bascuñán P;Niño-Garcia JP;Galeano-Castañeda Y;Serre D;Correa MM
通讯作者：
Correa MM
影响因子：
48
作者：
Callahan BJ;McMurdie PJ;Rosen MJ;Han AW;Johnson AJ;Holmes SP
通讯作者：
Holmes SP
影响因子：
4.6
作者：
Choo JM;Leong LE;Rogers GB
通讯作者：
Rogers GB
DOI：
10.1073/pnas.1412984112
发表时间：
2015-01-13
影响因子：
11.1
作者：
Carissimo, Guillaume;Pondeville, Emilie;Vernick, Kenneth D.
通讯作者：
Vernick, Kenneth D.