四环素降解菌共培养体系构建及废水修复的群落分析

doi:10.13560/j.cnki.biotech.bull.1985.2020-0232

摘要/Abstract

摘要：

含四环素废水在环境中广泛存在,由于其会造成抗性基因传播及人体耐药性等问题而备受关注。通过比较菌株对四环素的生物降解性能,明确地构建了以菌株Raoultella sp. XY-1及Pandoraea sp. XY-2为主体的共培养系统,并将该系统引入实际废水进行生物修复,观测污染物去除效率及群落变化。结果显示在含80 mg/L四环素的葡萄糖培养基中,共培养微生物10 d降解率达到80%以上。共培养微生物对真实四环素废水污染物去除效果良好,且高通量测序结果显示菌株Raoultella sp. XY-1成为优势菌。

关键词: 四环素废水, 生物修复, 微生物共培养, 细菌群落

Abstract:

Tetracycline-containing wastewater widely exists in environment,and it has attracted much attention due to issues such as resistance gene transmission,human drug resistance,etc. After comparing the ability of bacteria degrading tetracycline,the co-culture system mainly with strain Raoultella sp. XY-1 and strain Pandoraea sp. XY-2 was clearly constructed. Then this system was applied in actual wastewater for bioremediation,and the removal efficiency of pollutants and bacterial community changes were recorded. The results showed that degradation percentage by this co-culture system for 10 d was > 80% in glucose medium with 80 mg/L tetracycline. The co-culture system presents a favorable effect on the removal of pollutants from actual tetracycline-containing wastewater,and the results by high-through sequencing community indicated that the strain Raoultella sp. XY-1 became the dominant bacterium.

Key words: tetracycline wastewater, bioremediation, microbial co-culture, bacteria community

吴学玲, 周翔宇, 吴晓燕, 罗奎, 顾怡超, 周晗, 廖婉晴, 曾伟民. 四环素降解菌共培养体系构建及废水修复的群落分析[J]. 生物技术通报, 2020, 36(10): 116-126.

WU Xue-ling, ZHOU Xiang-yu, WU Xiao-yan, LUO Kui, GU Yi-chao, ZHOU Han, LIAO Wan-qing, ZENG Wei-min. Construction of Tetracycline-degrading Bacterial Co-culture System and Community Analysis of Wastewater Remediation[J]. Biotechnology Bulletin, 2020, 36(10): 116-126.

图/表 7

表1 四环素回收率情况

添加浓度/ （mg?L^-1）	回收率/100%			${x}^{-}±SD \%$	CV/100%
添加浓度/ （mg?L^-1）	1	2	3	${x}^{-}±SD \%$	CV/100%
10	87.01	88.23	89.06	88.14±0.84	0.95
40	86.04	87.68	89.52	87.75±1.42	1.61

表2 不同时间样本显著差异分析

样本	对照组	Raoultella sp. XY-1	Pandoraea sp. XY-2	共培养系统	时间/d	显著性
四环素浓度	80.83±1.05a	80.38±0.55a	81.77±1.59a	79.77±0.70a	0	0.112
四环素浓度	65.90±0.74a	19.1±0.07c	28.94±0.32b	13.91±0.08d	12	1.000

图1 葡萄糖培养基中四环素残留浓度变化（A）和共培养体系比例优化（B）

表3 生物修复中理化参数变化情况

检测项目	空白	对照组	实验组	检测项目	空白	对照组	实验组
四环素浓度	3.34278	2.23645	1.03205	pH	6.82	7.04	8.31
COD_Cr	462.16	386.56	224.65		NH₄⁺-N	3.92	3.66	2.03
总氮	27.45	23.22	12.33		NO₃^--N	18.32	12.64	4.96
Ag	<0.001	<0.001	<0.001		As	0.031	0.011	<0.005
Cd	0.006	0.002	<0.001		Co	0.015	0.004	<0.001
Cr	0.013	0.009	<0.002		Cu	0.311	0.201	0.152
Fe	0.587	0.392	<0.005		Hg	0.007	0.003	0.003
Mn	<0.001	<0.001	<0.001		Ni	0.015	0.006	0.003
Pb	<0.009	<0.009	<0.009		Zn	0.321	0.154	<0.001

图2 实验组与对照组修复前后群落香浓指数分析（A）及实验组与对照组修复前后群落Chao指数分析（B）

图3 实验组与对照组修复前后所有属的重叠情况（A）,实验组与对照组修复前后纲水平群落丰度柱状图（B）,实验组与对照组修复前后群落属水平可视化圈图（C）

图4 对照组（A）实验组（B）与修复后的实验组与对照组（C）组内显著差异菌属 Y轴表示属水平下的物种名,物种对应的每个柱子表示该物种在各样本相对丰度,不同颜色表示不同的样本来源;中间区域为所设定的置信区间内,圆点对应的数值表示物种在两样本中相对丰度的差值,圆点颜色显示为物种丰度占比较大的样本颜色;图中最右边为P值,* 0.01 < P ≤ 0.05,** 0.001 < P ≤ 0.01,*** P ≤ 0.001

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