来自海洋海草 Streptomyces argenteolus TMA13 的生物活性化合物:利用时间致死动力学和活死细胞成像技术对抗鱼类病原体。

IF 2.1 4区 生物学 Q3 MICROBIOLOGY Brazilian Journal of Microbiology Pub Date : 2024-09-01 Epub Date: 2024-07-19 DOI:10.1007/s42770-024-01407-w
Lokesh Elumalai, Siddharthan Nagarajan, Sivarajan Anbalmani, Sangeetha Murthy, Radhakrishnan Manikkam, Balagurunathan Ramasamy
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引用次数: 0

摘要

放线菌普遍存在于水生和陆生环境中,呈丝状形态,拥有特定 G + C 含量的 DNA,并产生大量次级代谢产物。本研究侧重于从海洋海草中分离出的放线菌,研究它们对鱼类病原体的抗菌活性。采用琼脂栓塞法,在 28 个分离菌株中,银色链霉菌 TMA13 对鱼类病原体--嗜水气单胞菌(10 毫米)、鱼腥单胞菌(22 毫米)、塔尔达爱德华氏菌(17 毫米)、哈维弧菌(22 毫米)和鳗弧菌(12 毫米)的抑制面积最大。这种强效菌株的优化涉及各种因素,包括 pH 值、温度、碳源和盐条件,以提高产量和抗菌效果。在抗生物膜试验中,随着 TMA13 提取物浓度的增加,抑菌率也随之增加。用 TMA13 粗提取物进行的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)检测表明,该提取物在极低浓度下对鱼类病原体具有很强的活性。时间杀灭动力学检测显示,所有鱼类病原体经 100 µg/ml 浓度的粗提取物处理后,其生长曲线在不同时间间隔内均有变化,表明细胞活力下降并进入死亡阶段。此外,还研究了荧光显微镜下细菌细胞暴露于萃取物后发出绿色和红色荧光的情况,从而实现了活死细胞的分化。通过气相色谱-质谱(GC-MS)和傅立叶变换红外光谱(FT-IR)分析对粗萃取物进行了进一步表征,确定了具有显著抗菌活性的功能基团次生代谢物。这项研究阐明了阿根廷链霉菌 TMA13 生产抗生素化合物的能力,可有效对抗鱼类病原体。
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Bioactive compound from marine seagrass Streptomyces argenteolus TMA13: combatting fish pathogens with time-kill kinetics and live-dead cell imaging.

Actinobacteria, pervasive in aquatic and terrestrial environments, exhibit a filamentous morphology, possess DNA with a specific G + C content and production of numerous secondary metabolites. This study, focused on actinobacteria isolated from marine seagrass, investigating their antibacterial activity against fish pathogens. Among 28 isolates, Streptomyces argenteolus TMA13 displayed the maximum zone of inhibition against fish pathogens-Aeromonas hydrophila (10 mm), Aeromonas caviae (22 mm), Edwardsiella tarda (17 mm), Vibrio harveyi (22 mm) and Vibrio anguillarum (12 mm) using the agar plug method. Optimization of this potent strain involved with various factors, including pH, temperature, carbon source and salt condition to enhance both yield production and antibacterial efficacy. In anti-biofilm assay shows the maximum percentage of inhibition while increasing concentration of TMA13 extract. Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) assays with TMA13 crude extract demonstrated potent activity against fish pathogens at remarkably low concentrations. Time-kill kinetics assay showcased growth curve variations over different time intervals for all fish pathogens treated with a 100 µg/ml concentration of crude extract, indicating a decline in cells viability and progression into the death phase. Additionally, fluorescence microscopic visualization of bacterial cells exposed to the extracts emitting green and red fluorescence, enabling live-dead cell differentiation was also studied. Further characterization of the crude extract through GC-MS and FT-IR analyses performed and identified secondary metabolites with functional groups exhibiting significant antibacterial activity. This study elucidates the capacity of Streptomyces argenteolus TMA13 to enhance the production of antibiotic compounds effective against fish pathogens.

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来源期刊
Brazilian Journal of Microbiology
Brazilian Journal of Microbiology 生物-微生物学
CiteScore
4.10
自引率
4.50%
发文量
216
审稿时长
1.0 months
期刊介绍: The Brazilian Journal of Microbiology is an international peer reviewed journal that covers a wide-range of research on fundamental and applied aspects of microbiology. The journal considers for publication original research articles, short communications, reviews, and letters to the editor, that may be submitted to the following sections: Biotechnology and Industrial Microbiology, Food Microbiology, Bacterial and Fungal Pathogenesis, Clinical Microbiology, Environmental Microbiology, Veterinary Microbiology, Fungal and Bacterial Physiology, Bacterial, Fungal and Virus Molecular Biology, Education in Microbiology. For more details on each section, please check out the instructions for authors. The journal is the official publication of the Brazilian Society of Microbiology and currently publishes 4 issues per year.
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