创伤弧菌生物被膜形成能力及影响因素的研究

高梦迪; 宁喜斌

doi:10.13386/j.issn1002-0306.2022080256

创伤弧菌生物被膜形成能力及影响因素的研究

doi: 10.13386/j.issn1002-0306.2022080256

高梦迪^1,,
宁喜斌^{1, 2, 3,*, ,}

1.
上海海洋大学食品学院，上海 201306
2.
农业部水产品贮藏保鲜质量安全风险评估试验室（上海），上海 201306
3.
国家淡水水产品加工技术研发分中心（上海），上海 201306

详细信息

作者简介:
高梦迪（1996−），女，硕士研究生，研究方向：食品微生物，E-mail：1404843089@qq.com

通讯作者:
宁喜斌（1964−），男，博士，教授，研究方向：微生物学、食品安全，E-mail：xbning@shou.edu.cn

中图分类号: TS201.3
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- 文章访问数: 17
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- 被引次数: 0
出版历程
- 收稿日期: 2022-08-24
- 刊出日期: 2023-05-15

Biofilm Forming Ability and Influencing Factors of Vibrio vulnificus

GAO Mengdi^1
,,
NING Xibin^{1, 2, 3,*
, ,}

1.
College of Food Science and Technology, Shanghai Ocean University, Shanghai 201306, China
2.
Laboratory of Quality and Safety Risk Assessment for Aquatic Products on Storage and Preservation (Shanghai), Ministry of Agriculture, Shanghai 201306, China
3.
National R&D Branch Center for Freshwater Aquatic Products Processing Technology (Shanghai), Shanghai 201306, China

摘要

摘要: 研究26株创伤弧菌（Vibrio vulnificus，Vv）产生物被膜情况及影响生物被膜形成的因素，为有效控制创伤弧菌形成生物被膜提供理论依据。本研究采用刚果红平板法、改良试管法及改良微孔板法分析25株创伤弧菌分离株及1株标准菌株形成生物被膜的能力，从中选出一株产膜能力最强的菌株，并研究不同初始菌浓度、温度及时间、pH、NaCl浓度、金属阳离子以及接触材料对其生物被膜形成的影响。结果显示，所选菌株中具有生物被膜形成能力的有25株（96.15%）。其中菌株VvK产膜能力最强，在25 ℃条件下，初始菌浓度为10⁸ CFU/mL，含3% NaCl、pH8~9培养24 h时，生物被膜形成量最大。而添加一定浓度的金属阳离子（Cu²⁺、Mn²⁺、Ca²⁺、Mg²⁺）后，生物被膜的形成受到不同程度的抑制，其抑制能力依次降低。菌株VvK在接触亲水性表面（不锈钢和玻璃）时生物被膜形成量显著高于疏水性表面（聚苯乙烯），且在不锈钢表面形成量最大。不同创伤弧菌生物被膜形成能力具有较大的差异，且在不同培养条件下具有特定的规律，需引起重点关注。
- 创伤弧菌 /
- 生物被膜 /
- 培养条件 /
- 影响因素 /
- 产膜能力
Abstract: The biofilm produced by 26 strains of Vibrio vulnificus (Vv) and the factors affecting biofilm formation were studied to provide theoretical basis for effective control of biofilm formation by Vv. In this study, Congo red plate method, improved test tube method and improved microplate method were used to analyze the ability of 25 isolated Vv strains and 1 standard strain to form biofilm, and one strain with the strongest ability to produce biofilm was selected from them. The effects of different initial bacterial concentration, temperature, culture time, pH, NaCl concentration, metal cations and contact materials on the formation of biofilm were investigated. The results showed that 25 strains (96.15%) had biofilm forming ability among the selected strains and VvK had the strongest ability to produce biofilm. At 25 ℃, the initial bacterial concentration was 10⁸ CFU/mL, 3% NaCl and pH8~9 for 24 h, the biofilm quantity was the largest. However, the formation of biofilm was inhibited by certain concentration of metal cations (Cu²⁺, Mn²⁺, Ca²⁺, Mg²⁺) definitely, and the inhibition ability of above cations decreases in turn. When VvK contacted hydrophilic surfaces (stainless steel and glass), the amount of biofilm formation was significantly higher than hydrophobic surfaces (polystyrene), and the amount of biofilm formation on stainless steel was the largest. The biofilm forming ability of different Vv was quite different, and had specific rules under different cultivation conditions, which should be paid more attention.
- Vibrio vulnificus /
- biofilm /
- culture conditions /
- influence factor /
- film production capacity

HTML全文

图 1 刚果红平板鉴定生物被膜

注：左：PIA阳性菌株VvK；右：PIA阴性菌株Vv Bq。

Figure 1. Congo red plate for biofilm identification

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图 2 改良试管法鉴定生物被膜

注：第3排第1管为对照管，其余为试验管。

Figure 2. Biofilm identification by modified tube method

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图 3 初始菌浓度对VvK生物被膜的影响

Figure 3. Effect of initial concentration on VvK biofilm

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图 4 NaCl浓度对VvK生物被膜的影响

Figure 4. Effect of NaCl concentration on VvK biofilm

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图 5 pH对VvK生物被膜的影响

注：不同小写字母表示差异显著（P<0.05），相同小写字母表示差异不显著（P>0.05）。

Figure 5. Effect of pH on VvK biofilm

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图 6 温度和时间对VvK生物被膜的影响

Figure 6. Effects of temperature and time on VvK biofilm

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图 7 金属阳离子对VvK生物被膜的影响

Figure 7. Effect of metal cations on VvK biofilm

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图 8 不同接触材料对VvK生物被膜的影响

Figure 8. Effects of different contact materials on VvK biofilm

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表 1 创伤弧菌生物被膜测定结果

Table 1. Results of Vibrio vulnificus biofilm determination

生物被膜粘附程度	测定值范围（OD_570nm）	菌株数目（占比%）
+++	0.477±0.008～1.877±0.035	9（34.6%）
++	0.322±0.032～0.466±0.018	13（50%）
+	0.214±0.007～0.230±0.020	3（11.6%）
−	0.088±0.009	1（3.8%）
	0.088±0.009	空白对照
注：+++：强粘附，++：中等粘附，+：弱粘附，−：无粘附。

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表 2 强生物被膜形成能力菌株

Table 2. Strong ability of biofilm formation strains

菌株名称	OD_570nm值（平均值±标准差）
VvK	1.877±0.035^a
Vv 18s	0.658±0.022^b
Vv 61	0.656±0.026^b
Vv Ds	0.572±0.005^c
Vv Cs	0.519±0.038^d
Vv Js	0.518±0.030^d
Vv 52	0.513±0.019^d
Vv Y5	0.503±0.023^d
Vv CICC21615	0.477±0.009^d
注：不同小写字母表示差异显著（P<0.05），相同小写字母表示差异不显著（P>0.05）。

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