Inhibition of citral nanoemulsion to growth,spoilage ability and Al-2/luxS quorum sensing system of Shewanella putrefaciens CN-32:a study on bacteriostasis from in vitro culture and gene expression analysis

Objectives:The bacteriostatic effects of a citral nanoemulsion against Shewanella putrefaciens CN-32(SHP CN-32)were investigated using in vitro culture and gene expression analysis,forbuilding a potential application in spoilage microorganism control and aquatic products quality maintenance.Materials and Methods:SHP CN-32 was treated by prepared citral nanoemulsion when the minimal inhibitory concentration(MIC)was verified.The growth curve,membrane integrity,scanning electron microscope(SEM)observation,biofilm formation and quorum sensing(QS)signaling molecule Al-2 content were evaluated in different MIC treatment groups(0 to 1.00 MIC).The gene expression status of SHP CN-32 in O and 0.50 MIC groups were compared using transcriptome sequencing and quantitative polymerase chain reaction(PCR).Results:The in vitro culture revealed that the citral nanoemulsion could inhibit the growth of SHP CN-32 with MIC of approximately 200μg/mL.Images of membrane integrity.SEM and biofilm formation suggested significant biological structure damage in bacteria after treatment.Meanwhile,the Qs signaling molecule Al-2 content showed a decline with increasing treatment concentration.Transcriptome sequencing and quantitative PCR revealed that the majority genes related diversified functional metabolic pathways of SHP CN-32 were downregulated at varying degree.Conclusion:A significant bacteriostasis of citral nanoemulsion against SHP CN-32 was verified via the results of growth inhibition,structural destruction,signal molecular decrease and gene expression downregulation of strains.These synergies significantly affect the characteristic expression of SHP CN-32,revealing the application potential as bacteriostat,QS inhibitor and preservative in aquatic products.