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中国海水养殖系统中分离的芽孢杆菌属的脱氮特性及时空差异生态风险评估

The nitrogen removal characterization and ecological risk assessment of Bacillus sp. isolated from mariculture systems in China with spatiotemporal difference.

作者信息

Zhang Qian, Wang Yingeng, Zhang Zheng, Yu Yongxiang, Wang Chunyuan, Liao Meijie, Rong Xiaojun, Zhang Zhiqi, Li Bin, Ge Jianlong, Wang Jinjin

机构信息

School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao, Shandong, China.

State Key Laboratory of Mariculture Biobreeding and Sustainable Goods, Yellow Sea Fisheries Research Institute, Chinese Academic of Fishery Sciences, Qingdao, Shandong, China.

出版信息

PLoS One. 2025 Mar 20;20(3):e0319344. doi: 10.1371/journal.pone.0319344. eCollection 2025.

Abstract

The accumulation of nitrogen compounds may worsen the aquatic environment and cause serious economic losses in the aquaculture industry. In this study, the denitrification performance and ecological safety of 120 Bacillus sp. isolates with spatial and temporal differences were evaluated based on the aspects of hemolysis, drug resistance, denitrification performance, and purification effect for mariculture wastewater. Firstly, 55/120 safe strains with no hemolytic activity were detected through hemolysis testing. Then, based on selective denitrification medium and colorimetric reagent method, 34/55 Bacillus sp. with denitrification effect were screened. For these 34 Bacillus sp. isolates, the drug resistance phenotype and genotype, denitrification genes, and enzyme activities related to the nitrogen metabolizing (AMO, HAO, NAR, NIR) were examined. And the MARI was 0.00-0.25, with a multi-drug resistance rate of 17.6%. The drug resistance genes tetB, blaTEM, and cfr and the denitrification genes nap, nor, and narG were detected. Ultimately, 27/34 strains with denitrification function and ecological safety were obtained. In addition, eight Bacillus sp. showed certain denitrification effects on nitrogen-containing wastewater treatment. Among them, B. subtilis B24 has outstanding denitrification ability, with removal rates of 92%, 62%, 68%, and 30% for NH4 + -N, NO2--N, NO3--N, and TN in simulated wastewater, respectively. It also has a good denitrification effect in practical applications. This study provides candidate bacterial strains for the treatment of mariculture wastewater.

摘要

氮化合物的积累可能会使水生环境恶化,并给水产养殖业造成严重的经济损失。在本研究中,基于溶血、耐药性、反硝化性能以及对海水养殖废水的净化效果等方面,对120株芽孢杆菌属菌株的时空差异反硝化性能及生态安全性进行了评估。首先,通过溶血试验检测出55株/120株无溶血活性的安全菌株。然后,基于选择性反硝化培养基和比色试剂法,筛选出34株/55株具有反硝化效果的芽孢杆菌属菌株。对于这34株芽孢杆菌属菌株,检测了其耐药表型和基因型、反硝化基因以及与氮代谢相关的酶活性(AMO、HAO、NAR、NIR)。MARI为0.00 - 0.25,多重耐药率为17.6%。检测到耐药基因tetB、blaTEM和cfr以及反硝化基因nap、nor和narG。最终,获得了27株/34株具有反硝化功能且生态安全的菌株。此外,8株芽孢杆菌属菌株对含氮废水处理表现出一定的反硝化效果。其中,枯草芽孢杆菌B24具有出色的反硝化能力,对模拟废水中的NH4 + -N、NO2--N、NO3--N和TN的去除率分别为92%、62%、68%和30%。其在实际应用中也具有良好的反硝化效果。本研究为海水养殖废水处理提供了候选菌株。

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