中国农业科学 ›› 2019, Vol. 52 ›› Issue (9): 1614-1623.doi: 10.3864/j.issn.0578-1752.2019.09.012
蔡林林1,胡海静1,衣晓坤2,王虎虎1(),徐幸莲1,彭斌3
收稿日期:
2018-09-04
接受日期:
2019-01-31
出版日期:
2019-05-01
发布日期:
2019-05-16
通讯作者:
王虎虎
作者简介:
蔡林林,E-mail: 基金资助:
CAI LinLin1,HU HaiJing1,YI XiaoKun2,WANG HuHu1(),XU XingLian1,PENG Bin3
Received:
2018-09-04
Accepted:
2019-01-31
Online:
2019-05-01
Published:
2019-05-16
Contact:
HuHu WANG
摘要:
【目的】研究酸性电解水(AEW)能否有效控制肉源性荧光假单胞菌,探究处理后菌体变化,为新型杀菌剂的开发提供思路。【方法】使用不同有效氯浓度酸性电解水(20、40和60 mg?L -1)将肉源性荧光假单胞菌分别处理5、10、15、20、25和30 min后,通过平板计数检测酸性电解水对肉源性荧光假单胞菌的致死效果;收集未处理及20、40和60 mg?L -1酸性电解水各处理5 min后菌液,利用扫描电子显微镜观测酸性电解水对肉源性荧光假单胞菌细胞形态的影响;利用多种荧光探针检测酸性电解水对肉源性荧光假单胞菌细胞膜完整性、胞内pH、胞内ATP浓度、膜电位的影响;提取胞外聚合物(EPS)探究酸性电解水与EPS的相互作用。 【结果】20、40和60 mg?L -1的酸性电解水均可在5 min内杀死10 7CFU/mL肉源性荧光假单胞菌,酸性电解水浓度大于40 mg·L -1时对肉源性荧光假单胞菌的杀菌效果没有显著影响(P>0.05),10 min处理后均已无活菌检出。扫描电子显微镜结果表明3种浓度酸性电解水的处理均可使肉源性荧光假单胞菌菌体干瘪皱缩,失去原本饱满的细胞形态,60 mg·L -1电解水处理下破损程度最大。酸性电解水可显著降低肉源性荧光假单胞菌的膜完整性、胞内pH、膜电位、胞内ATP浓度和EPS含量(P<0.05)。20、40和60 mg·L -1酸性电解水处理后菌体膜完整性分别降低到2.26%、1.87%和1.20%,膜电位分别消散40%、50%和50%,胞内pH由7.50分别降低到6.53、5.90和5.83,3种浓度处理后胞内ATP浓度已降至最低。杀菌效果显著依赖于酸性电解水浓度的升高,当处理浓度高于40 mg?L -1时,胞内pH、膜电位和胞内ATP浓度的降低无显著差异。酸性电解水处理下EPS中蛋白、总糖含量显著降低,松散胞外聚合物 (L-EPS)中蛋白和总糖分别降低71.08%和62.23%,而紧密胞外聚合物(B-EPS)中蛋白和总糖分别降低99.32%和40.62%。当EPS反向添加到酸性电解水中时则可显著降低酸性电解水氧化还原电位(ORP)(P<0.05),但对有效氯浓度(ACC)和pH没有显著影响(P>0.05)。 【结论】酸性电解水对肉源性荧光假单胞菌具有良好的杀菌效果,可破坏肉源性荧光假单胞菌细胞膜,同时EPS对酸性电解水杀菌效果具有一定的阻碍作用。
蔡林林, 胡海静, 衣晓坤, 王虎虎, 徐幸莲, 彭斌. 酸性电解水对肉源性荧光假单胞菌的致死效应[J]. 中国农业科学, 2019, 52(9): 1614-1623.
CAI LinLin, HU HaiJing, YI XiaoKun, WANG HuHu, XU XingLian, PENG Bin. Lethal Effect of Meat-Borne Pseudomonas fluorescens to Acidic Electrolyzed Water[J]. Scientia Agricultura Sinica, 2019, 52(9): 1614-1623.
表2
酸性电解水对肉源性荧光假单胞菌胞外聚合物中蛋白和总糖含量影响"
类型 Type | 处理 Treatment | 蛋白 Protein (μg?mg -1) | 总糖 Carbohydrate (μg?mg -1) | 蛋白/总糖 Protein/Carbohydrate ratio |
---|---|---|---|---|
松散胞外聚合物L-EPS | 0 | 583.68±72.05a | 112.33±0.73a | 5.20±0.18a |
40 mg?L-1 AEW | 168.78±2.96b | 42.36±0.14b | 3.98±0.08b | |
紧密胞外聚合物B-EPS | 0 | 59.23±1.28a | 19.72±0.66a | 3.01±0.15a |
40 mg?L-1 AEW | 0.40±0.28b | 11.71±1.23b | 0.03±0.02b |
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