Kydd LeNaiya, Alalhareth Fawaz, Mendez Ana, Hohn Maryann, Radunskaya Ami, Kojouharov Hristo, Jaworski Justyn
Department of Bioengineering, The University of Texas at Arlington, Arlington, TX 76010 USA.
Department of Mathematics, The University of Texas at Arlington, Arlington, TX 76010 USA.
Regen Eng Transl Med. 2022;8(3):489-497. doi: 10.1007/s40883-022-00248-z. Epub 2022 Apr 14.
We examine the impacts of dosing strategies of plasmids on bacterial communities in the murine gut by measuring the quantity of plasmids in mouse feces.
We fed mice carrier bacteria, , that contain plasmids with both a reporter gene and an antibiotic resistant gene. We varied the quantity of the plasmid-carrying bacteria and the length of time the mice consumed the bacteria. We also pretreated the gut with broad-spectrum antibiotics and used continuous antibiotic treatment to investigate selection pressure. We collected bacteria from fecal pellets to quantify the number of plasmid-carrying bacteria via plate assay.
Dosing regimens with plasmid-carrying bacteria resulted in a significantly increased duration of persistence of the plasmid within the gut when supplemented continuously with kanamycin during as well as after completion of bacterial dosing. The carrier bacteria concentration influenced the short-term abundance of carrier bacteria.
We evaluated the persistence of plasmid-carrying bacteria in the murine gut over time using varying dosage strategies. In future work, we will study how bacterial diversity in the gut impacts the degree of plasmid transfer and the prevalence of plasmid-carrying bacteria over time.
Observing how plasmids persist within the gut can help us understand how newly introduced genes, including antibiotic resistance, are transmitted within the gut microbiome. In our experiments, mice were given bacteria containing a genetically engineered plasmid and were examined for the persistence of the plasmid in the gut. We found long-term persistence of the plasmid in the gut when administering antibiotics during and following dosing of the mice with bacteria carrying the plasmid. The use of higher concentrations of carrier bacteria influenced the short-term abundance of the plasmid-carrying bacteria in the gut.
Building on evidence from these initial studies that persistence of plasmids within the gut can be regulated by the dosage strategy, we will explore future studies and models of gene uptake in the context of spatial and taxonomic control and further determine if dosing strategies alter the compositional diversity of the gut microbiome.
通过测量小鼠粪便中质粒的数量，研究质粒给药策略对小鼠肠道细菌群落的影响。
我们给小鼠喂食携带质粒的载体细菌，该质粒同时含有报告基因和抗生素抗性基因。我们改变携带质粒细菌的数量以及小鼠摄入细菌的时间长度。我们还用广谱抗生素对肠道进行预处理，并采用持续抗生素治疗来研究选择压力。我们从粪便颗粒中收集细菌，通过平板试验来量化携带质粒细菌的数量。
当在细菌给药期间以及给药完成后持续补充卡那霉素时，携带质粒细菌的给药方案导致质粒在肠道内的持续时间显著增加。载体细菌浓度影响载体细菌的短期丰度。
我们使用不同的给药策略评估了携带质粒细菌在小鼠肠道内随时间的持续性。在未来的工作中，我们将研究肠道细菌多样性如何随时间影响质粒转移程度和携带质粒细菌的流行率。
观察质粒如何在肠道内持续存在有助于我们理解新引入的基因，包括抗生素抗性基因，如何在肠道微生物群中传播。在我们的实验中，给小鼠喂食含有基因工程质粒的细菌，并检测质粒在肠道内的持续性。我们发现，在给携带质粒的细菌给小鼠给药期间及之后使用抗生素时，质粒在肠道内长期持续存在。使用更高浓度的载体细菌会影响肠道内携带质粒细菌的短期丰度。
基于这些初步研究的证据，即质粒在肠道内的持续性可通过给药策略进行调节，我们将在空间和分类控制的背景下探索未来的基因摄取研究和模型，并进一步确定给药策略是否会改变肠道微生物群的组成多样性。
