姜黄素介导的光动力技术对牡蛎汁中副溶血性弧菌生物被膜的清除

Inactivation effects of curcumin-mediated photodynamic technology on Vibrio parahaemolyticus biofilm formed in oyster juice

副溶血性弧菌是水产品中常见的食源性致病菌,该研究以水产品中常见的牡蛎为研究对象,建立了牡蛎汁中副溶血性弧菌生物被膜的生长模型,并采用新型高效的姜黄素介导的光动力灭活(photodynamic inactivation,PDI)技术以清除牡蛎汁中形成的成熟副溶血性弧菌生物被膜。通过结晶紫染色、噻唑蓝[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumromide,MTT]法研究副溶血性弧菌在牡蛎汁以及胰蛋白胨大豆肉汤(trypticase soy broth,TSB)培养基中生物被膜形成能力和细胞活力的差异,并探究PDI处理副溶血性弧菌生物被膜前后的生物被膜量、基因表达水平、细胞形态和被膜结构参数的变化。结果表明,与TSB培养基相比,在牡蛎汁中能够形成细胞活力更强的生物被膜,并且在姜黄素浓度为20μmol/L、蓝光照射60 min(13.68 J/cm 2)条件下,PDI处理对牡蛎汁中形成的副溶血性弧菌生物被膜量清除率为>83%。此外,PDI处理还下调了氧化应激相关基因(grx A、sod C)和SOS反应基因(lex A),破坏了生物被膜的三维结构,其生物体积、厚度和质构熵降低,均一性增加。以上结果表明,姜黄素介导的光动力具有良好地清除牡蛎中副溶血性弧菌生物被膜的效果,该研究为预防副溶血性弧菌引起的食源性疾病及水产品保鲜提供了借鉴方法与思路。

Vibrio parahaemolyticus represents a common pathogenic bacterium found in aquatic products.In this study,we selected oysters as our research model and established a growth model for V.parahaemolyticus biofilm in oyster juice.We further introduced a novel curcumin-mediated photodynamic inactivation(PDI)approach to eliminate the mature V.parahaemolyticus biofilm present in the oyster juice.Using crystal violet staining and MTT assays,we assessed the capacity for biofilm formation and cell viability in both oyster juice and TSB medium.Subsequent investigations focused on quantifying biofilm biomass,elucidating gene expression levels,and characterizing cell morphology as well as biofilm structural parameters.Our findings revealed a more robust biofilm formation in oyster juice as compared to the TSB medium.Remarkably,over 83%of the V.parahaemolyticus biofilm in oyster juice was eradicated after PDI treatment using a concentration of 20.0μmol/L curcumin and 60 min(13.68 J/cm 2)blue LED irradiation.Notably,the PDI treatment downregulated oxidative stress-related genes(grx A and sod C)and SOS response genes(lex A)within the biofilm cells.This treatment also compromised the three-dimensional architecture of the biofilm,with evident reductions in bio-volume,thickness,and textural entropy,while concurrently enhancing its homogeneity.This approach offers a novel method for mitigating foodborne diseases caused by V.parahaemolyticus and improving the preservation of aquatic products.