Snoeyenbos-West Oona L O, Guerrero Christina R, Valencia Makaela, Carini Paul
Department of Environmental Science, University of Arizona, Tucson, AZ 85721.
School of Animal and Comparative Biomedical Science, University of Arizona, Tucson, AZ 85721.
bioRxiv. 2023 Oct 10:2023.10.10.561742. doi: 10.1101/2023.10.10.561742.
Anaerobic microbes play crucial roles in environmental processes, industry, and human health. Traditional methods for monitoring the growth of anaerobes, including plate counts or subsampling broth cultures for optical density measurements, are time and resource intensive. The advent of microplate readers revolutionized bacterial growth studies by enabling high-throughput and real-time monitoring of microbial growth kinetics but their use in anaerobic microbiology has remained limited. Here, we present a workflow for using small-footprint microplate readers and the Growthcurver R package to analyze the kinetic growth metrics of anaerobic bacteria. We benchmarked the small-footprint Cerillo Stratus microplate reader against a BioTek Synergy HTX microplate reader in aerobic conditions using DSM 28618 cultures. The growth rates and carrying capacities obtained from the two readers were statistically indistinguishable. However, the area under the logistic curve was significantly higher in cultures monitored by the Stratus reader. We used the Stratus to quantify the growth responses of anaerobically grown and DSM 29485 to different doses of the toxin sodium arsenite. The growth of and was sensitive to arsenite doses of 1.3 μM and 0.4 μM, respectively. Complete inhibition of growth was achieved at 38 μM arsenite for , and 338 μM in . These results show that the Stratus performs similarly to a leading brand of microplate reader and can be reliably used in anaerobic conditions. We discuss the advantages of the small format microplate readers and our experiences with the Stratus.
厌氧微生物在环境过程、工业和人类健康中发挥着关键作用。监测厌氧菌生长的传统方法，包括平板计数或对肉汤培养物进行二次取样以测量光密度，既耗时又耗费资源。酶标仪的出现彻底改变了细菌生长研究，它能够对微生物生长动力学进行高通量实时监测，但在厌氧微生物学中的应用仍然有限。在此，我们展示了一种使用小型酶标仪和Growthcurver R软件包分析厌氧细菌动力学生长指标的工作流程。我们在有氧条件下使用DSM 28618培养物，将小型Cerillo Stratus酶标仪与BioTek Synergy HTX酶标仪进行了对比测试。从这两种酶标仪获得的生长速率和承载能力在统计学上没有显著差异。然而，Stratus酶标仪监测的培养物中逻辑曲线下的面积明显更高。我们使用Stratus酶标仪来量化厌氧培养的[细菌名称1]和DSM 29485对不同剂量毒素亚砷酸钠的生长反应。[细菌名称1]和DSM 29485的生长分别对1.3 μM和0.4 μM的亚砷酸盐剂量敏感。对于[细菌名称1]，在38 μM亚砷酸盐时生长完全受到抑制，对于DSM 29485则在338 μM时完全抑制。这些结果表明，Stratus酶标仪的性能与领先品牌的酶标仪相似，并且可以在厌氧条件下可靠使用。我们讨论了小型酶标仪的优点以及我们使用Stratus酶标仪的经验。
