渔用氧化剂对水源水和池塘水中磺胺类抗性基因<i>sul</i>1的去除作用

赵晓雨; 苏浩昌; 徐煜; 徐武杰; 胡晓娟; 文国樑; 曹煜成; 余招龙

doi:10.12131/20200231

渔用氧化剂对水源水和池塘水中磺胺类抗性基因sul1的去除作用

doi: 10.12131/20200231

赵晓雨^{1, 2,},
苏浩昌^{2, 3},
徐煜^{2, 3},
徐武杰^{2, 3},
胡晓娟^{2, 3},
文国樑^{2, 3},
曹煜成^{2, 3, ,},
余招龙⁴

1.
上海海洋大学水产与生命学院，上海 201306
2.
中国水产科学研究院南海水产研究所/农业农村部南海渔业资源开发利用重点实验室/广东省渔业生态环境重点实验室，广东广州 510300
3.
中国水产科学研究院南海水产研究所深圳试验基地，广东深圳 518121
4.
广东冠利海洋生物有限责任公司，广东茂名 525400

基金项目: 国家重点研发计划项目 (2019YFD0900402)，中国水产科学研究院基本科研业务费专项资金 (2020TD54)；现代农业产业技术体系建设专项资金 (CARS-48)；广东省现代农业产业技术体系创新团队建设专项资金 (2019KJ149)

详细信息

作者简介:
赵晓雨 (1996—)，男，硕士研究生，研究方向为养殖环境安全。E-mail: 13576943750@163.com

通讯作者:
曹煜成 (1979—)，男，博士，特聘研究员，从事水产健康养殖和养殖生态环境调控研究。E-mail: cyc_715@163.com

中图分类号: S 949
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- 被引次数: 0
出版历程
- 收稿日期: 2020-11-05
- 修回日期: 2021-03-08
- 网络出版日期: 2021-04-13

Removal of sulphonamide resistance sul1 gene in water source and pond water by fishery oxidants in aquaculture

Xiaoyu ZHAO^{1, 2
,},
Haochang SU^{2, 3},
Yu XU^{2, 3},
Wujie XU^{2, 3},
Xiaojuan HU^{2, 3},
Guoliang WEN^{2, 3},
Yucheng CAO^{2, 3
, ,},
Zhaolong YU⁴

1.
College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China
2.
South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences/Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs/Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, Guangzhou 510300, China
3.
Shenzhen Base South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Shenzhen 518121, China
4.
Guangdong Guanli Marine Biology Co., Ltd., Maoming 525400, China

摘要

摘要: 为探讨利用渔用氧化剂去除养殖水体环境中的抗生素抗性基因 (Antibiotic resistant genes, ARGs) 并控制其传播的可行性，该研究以磺胺类抗性基因sul1作为目标抗性基因，选用次氯酸钠 (NaClO)、二溴海因 (C₅H₆Br₂N₂O₂) 和高锰酸钾 (KMnO₄) 3种养殖中常用的渔用氧化剂，运用实时荧光定量PCR技术，分析目标渔用氧化剂对水源水和池塘水体中ARGs的去除效果。结果显示，测试的水源水和池塘水体中的sul1浓度均处于较高水平，分别达到了10⁵ 拷贝·mL⁻¹和10⁶ 拷贝·mL⁻¹，而且微生物的细胞内sul1是其主要存在形式；其次，NaClO对各实验组中的sul1均呈现较强的去除效果，C₅H₆Br₂N₂O₂仅对水源水中细胞内和细胞外的sul1具有较强的去除作用，KMnO₄仅对不同水体中的细胞外sul1具有较强的去除作用。结果表明，3种目标渔用氧化剂中，NaClO对水源水和池塘水体中ARGs的去除效果最好，科学应用NaClO有利于去除水产养殖水环境中的ARGs，进而防控其在养殖过程中的传播。
- 抗生素抗性基因 /
- 渔用氧化剂 /
- 次氯酸钠 /
- 二溴海因 /
- 高锰酸钾
Abstract: To explore the feasibility of using fishery oxidants to eliminate antibiotic resistant genes (ARGs) in aquaculture water and to control their spread, we studied the effects on ARG removal of three commonly used fishery oxidants (Sodium hypochlorite, dibromohydantoin and potassium permanganate) in water source and pond water by real-time fluorescent quantitative PCR. Results show that the concentration of sul1 gene was high in the water source and pond water (10⁵ and 10⁶ copies·mL⁻¹, respectively), and the intracellular sul1 gene was the predominant form. Additionally, sodium hypochlorite had a strong effect on sul1 gene removal in all the experimental groups, and dibromohydantoin only showed a strong removal effect on sul1 gene in the water source. Potassium permanganate only showed a strong effect on the removal of extracellular sul1 gene in both water source and pond water. The results indicate that sodium hypochlorite has the best effect on ARG removal in both water source and pond water for these three common used fishery oxidants. Scientific application of sodium hypochlorite is beneficial to ARG removal and prevention in aquaticulture water.
- Antibiotic resistance genes /
- Fishery oxidants /
- Sodium hypochlorite /
- Dibromohydantoin /
- Potassium permanganate

HTML全文

图 1 消毒前后水源水中i-sul1(a)和e-sul1(b)浓度的变化

同一图中标有不同字母代表有显著差异 (P<0.05)；图2同此

Figure 1. Change in concentrations of i-sul1 (a) and e-sul1 (b) in water source before and after disinfection

Different letters in the same figure indicate significant difference (P<0.05). The same case in Figure 2.

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图 2 消毒前后池塘水中i-sul1 (a) 和e-sul1 (b) 浓度的变化

Figure 2. Change in concentrations of i-sul1 (a) and e-sul1 (b) in pond water before and after disinfection

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图 3 消毒过程中活性氯质量浓度 (a) 及其所达到的浓时积 (b) 的变化

Figure 3. Change in concentration of FAC (a) and reached CT values (b) during disinfection

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图 4 消毒过程中高锰酸钾质量浓度 (a) 及其所达到的浓时积 (b) 的变化

Figure 4. Change in concentration of KMnO₄ (a) and reached CT values (b) during disinfection

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表 1 水源水和池塘水中氨氮、亚硝酸盐氮和总有机碳质量浓度

Table 1. Concentrations of NH₄ ⁺-N, NO₂ ⁻-N and TOC in water source and pond water mg·L⁻¹

项目 Item	水源水 Water source	池塘水 Pond water
氨氮 NH₄ ⁺-N	0.127	1.120
亚硝酸盐氮 NO₂ ⁻-N	0.013	0.845
总有机碳 TOC	8.9	10.9

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表 2 qPCR所需引物

Table 2. Primers used for quantitative PCR

基因 Gene	引物对 Primer pair	序列 (5'—3') Sequence	退火温度 Annealing temperature/℃	片段大小 Amplicon size/bp	参考文献 Reference
sul1	FW	CGCACCGGAAACATCGCTGCAC	62	163	[23]
sul1	RV	TGAAGTTCCGCCGCAAGGCTCG	62	163	[23]
注：FW上游引物；RV下游引物 Note: FW. Forward primer; RV. Reverse primer

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