纳米银/聚乙烯醇复合物的生物合成及其对6种水产病原菌的抑菌活性

魏亚楠; 马新冉; 齐珈俪; 王先钰; 刘晓玲; 王磊

doi:10.13801/j.cnki.fhclxb.20210210.006

纳米银/聚乙烯醇复合物的生物合成及其对6种水产病原菌的抑菌活性

doi: 10.13801/j.cnki.fhclxb.20210210.006

鲁东大学　生命科学学院，烟台 264025

基金项目: 山东省重点研发计划(2019GSF107091)；山东省自然科学基金项目(ZR2020MC135)；山东省海洋与渔业科技创新计划(2017YY04)

详细信息

通讯作者:
王磊，博士，教授，研究方向为纳米技术在病害防控中的应用　E-mail：wanglei9909@163.com

共同作者：魏亚楠和马新冉为共同第一作者，对本文具有同等贡献
中图分类号: TB333
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出版历程
- 收稿日期: 2020-12-03
- 录用日期: 2021-02-05
- 网络出版日期: 2021-02-10
- 刊出日期: 2021-11-01

Biosynthesis of silver nanoparticles/polyvinyl alcohol composite and its antibacterial activity against six aquatic pathogens

College of Life Sciences, Ludong University, Yantai 264025, China

摘要

摘要: 纳米银作为一种新型抑菌剂有望成为传统抑菌剂的替代品，制备稳定、高效、环保的新型纳米银抑菌产品成为当今的研究热点。本研究以葡萄籽提取液为还原剂和稳定剂，聚乙烯醇(PVA)为载体，采用一步法“绿色”生物合成出一种纳米银/聚乙烯醇复合物(AgNPs/PVA)。通过紫外-可见(UV-Vis)吸收光谱、透射电镜(TEM)、X射线衍射(XRD)等手段对合成产物进行了表征。结果表明银离子被葡萄籽提取物成功还原成纳米银并附着在PVA的表面，纳米银颗粒均匀，呈现单分散状态，粒径较小，平均粒径为14 nm左右。AgNPs/PVA对鳗弧菌、溶藻弧菌、副溶血弧菌、哈维氏弧菌、灿烂弧菌及点状气单胞菌等6种典型的水产病原菌均有显著的抑菌效果。以溶藻弧菌为指示菌，AgNPs/PVA的最小抑菌浓度(MIC)为1.1 μg/mL，最小杀菌浓度(MBC)为2.2 μg/mL。AgNPs/PVA的Zeta电位为−24.1 mV，表明纳米银颗粒间有很强的排斥力，为其稳定分散提供保障，后续实验证明制备的AgNPs/PVA具有良好的稳定性和热稳定性。以上研究结果表明，AgNPs/PVA复合材料在水产养殖病害防治中具有广阔的应用前景。
- 纳米银 /
- 葡萄籽 /
- 聚乙烯醇 /
- 生物合成 /
- 抑菌活性 /
- 稳定性
Abstract: As a new generation of antibacterial agent, silver nanoparticles are expected to be a substitute for traditional antibacterial agents in the future. Developing a new stable, efficient and eco-friendly nano-silver antibacterial agent has become the hot spot of current research. In this study, the grape seeds extract was used as an reducing and stabilizing agent, and polyvinyl alcohol (PVA) was used as the carrier, the silver nanoparticles/polyvinyl alcohol (AgNPs/PVA) composites were prepared by a facile, green, “one-pot” biological method. The asprepared AgNPs/PVA composites were characterized by UV absorption spectroscopy (UV-Vis), transmission electron transmission electron microscopy (TEM) and X ray diffraction (XRD). The results indicate that the silver ions are successfully reduced by the grade seeds extract and the produced AgNPs are decorated on the surface of PVA. The AgNPs are uniform and monodisperse, the particle size is small with mean diameter about 14 nm. The AgNPs/PVA composites show superior antibacterial effects against six tipical aquatic pathogens, such as Vibrio splendidus, V. harveyi, V. anguillarum, V. alginolyticus, V. parahaemolyticus and Aeromonas punctata. They have a minimum inhibitory concentration (MIC) of 1.1 μg/mL and a minimum inhibitory concentration (MBC) of 2.2 μg/mL against V. alginolyticus. The Zeta potential of AgNPs/PVA is found to be −24.1 mV, indicating that the strong repulsion between the nanoparticles, which provide guarantee for stable dispersion. The subsequent experiments prove that the prepared AgNPs/PVA has good stability and thermostability. The above research results indicate that AgNPs/PVA composites have a great application prospect in the control of aquaculture diseases.
- silver nanoparticles /
- grape seeds /
- polyvinyl alcohol /
- biosynthesis /
- antibacterial activity /
- stability
注释:

1) 共同作者：魏亚楠和马新冉为共同第一作者，对本文具有同等贡献

HTML全文

图 1 (a)葡萄籽提取液、聚乙烯醇(PVA)和纳米银/聚乙烯醇复合物 (AgNPs/PVA)的照片；(b)葡萄籽提取液、PVA和AgNPs/PVA的紫外吸收光谱

1—Grape seeds extract; 2—PVA; 3—AgNPs/PVA

Figure 1. (a) Photograph of aqueous suspension of grape seeds extract, polyvinyl alcohol (PVA) and silver nanoparticles/polyvinyl alcohol (AgNPs/PVA) composites；(b) UV-Vis absorption spectra of grape seeds extract, PVA and AgNPs/PVA

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图 2 AgNPs/PVA透射电镜图像(a)与粒径分布图(b)

Figure 2. TEM images of AgNPs/PVA (a) and their particle size distributionsize (b)

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图 3 AgNPs/PVA的XRD图谱

Figure 3. X-ray diffraction pattern of AgNPs/PVA

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图 4 AgNPs/PVA对6种水产病原菌的抑菌圈实验

1—AgNPs/PVA; 2—AgNPs; 3—Grape seeds extract; 4—Normal saline

Figure 4. Inhibition zone test of 6 aquatic pathogens of AgNPs/PVA

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图 5 不同浓度AgNPs/PVA作用下溶藻弧菌的生长曲线

Figure 5. Growth curve of V. alginolyticus treated with different concentrations of AgNPs/PVA

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图 6 AgNPs/PVA对溶藻弧菌核酸内容物泄露的影响

Figure 6. Effects of AgNPs/PVA on the release of nucleic acid contents from V. alginolyticus

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图 7 AgNPs/PVA的Zeta电位

Figure 7. Zeta potential of AgNPs/PVA

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图 8 AgNPs/PVA热处理后的照片

1—20℃; 2—40℃; 3—60℃; 4—80℃; 5—100℃

Figure 8. Thermal stability test of AgNPs/PVA

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图 9 AgNPs/PVA的抑菌热稳定性实验

1—20℃; 2—40℃; 3—60℃; 4—80℃; 5—100℃ ；6—Grape seeds extract

Figure 9. Antibacterial thermal stability test of AgNPs/PVA

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表 1 AgNPs/PVA对6种水产致病菌的抑菌圈直径的统计结果(n=3)

Table 1. Statistics of the inhibition zones of 6 aquatic pathogenic bacteria of AgNPs/PVA (n=3)

Aquatic pathogens	Inhibition zone/mm
Aquatic pathogens	AgNPs/PVA	AgNPs	Grape seeds extract	Normal saline
V.splendidus	17.7±0.3	15.9±0.7	0	0
V. harveyi	17.6±0.2	16.2±0.2	0	0
V. anguillarum	20.3±0.4	17.2±0.3	0	0
V. alginolyticus	21.5±0.6	16.0±0.2	0	0
V. parahaemolyticus	19.3±0.1	17.1±0.4	0	0
A. Punctata	17.4±0.4	16.4±0.4	0	0

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表 2 AgNPs/PVA复合物对溶藻弧菌最小抑菌浓度和最小杀菌浓度的测定

Table 2. MIC and MBC tests of AgNPs/PVA composites against V. alginolyticus

Label	Concentration of AgNPs/PVA/(μg·mL⁻¹)	24 h	48 h
1	137.5	−	−
2	68.8	−	−
3	34.4	−	−
4	17.2	−	−
5	8.6	−	−
6	4.3	−	−
7	2.2	−	−
8	1.1	−	+
9	0.6	+	+
10	0.3	+	+
Negative control		−	−
Positive control		+	+

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表 3 葡萄籽生物合成AgNPs单体对溶藻弧菌最小抑菌浓度和最小杀菌浓度的测定

Table 3. MIC and MBC tests of AgNPs against V. alginolyticus

Label	Concentration of AgNPs/(μg·mL⁻¹)	24 h	48 h
1	65.5	−	−
2	32.8	−	−
3	16.4	−	−
4	8.2	−	−
5	4.1	−	−
6	2.0	−	−
7	1.0	−	+
8	0.5	+	+
9	0.2	+	+
10	0.1	+	+
Negative control		−	−
Positive control		+	+

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