二烯丙基二硫醚对蜂房哈夫尼菌群体感应的抑制作用
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摘要
以从海参中分离的优势腐败菌蜂房哈夫尼菌为对象,研究了二烯丙基二硫醚(DADS)对受其群体感应调控的生物膜及泳动性的抑制作用。采用CV026报告菌定性检测DADS的抗群体感应活性,利用96微孔板法、扫描电镜(SEM)和琼脂扩散法分析DADS对蜂房哈夫尼菌生物膜及泳动性的抑制作用。结果表明,DADS作用于蜂房哈夫尼菌和CV026的最低抑菌浓度是48 mmol/L。在亚抑菌浓度下,与未处理对照组相比,DADS能够抑制由群体感应调控的CV026中紫色素的分泌而不抑制其生长。在0～12 mmol/L,随着浓度增加,DADS对生物膜的抑制率可达49.6%,对泳动性的抑制率达到73.5%。在12 mmol/L时,DADS能够抑制生物膜的形成。DADS能够抑制蜂房哈夫尼菌的群体感应活性、生物膜及泳动性,从而发挥防腐功效。
The inhibitory effect of diallyl disulfide(DADS) on biofilm and swimming ability of Hafnia alvei, a dominant putrefaction bacterium isolated from sea cucumber, was studied. The anti-quorum sensing activity of DADS was qualitatively detected using CV026 as reporter bacteria. The inhibition of DADS on biofilm and swimming activity of Hafnia alvei were analyzed by 96-microplate method, scanning electron microscopy(SEM) and agar diffusion method. The results showed that the minimum inhibitory concentration of DADS on Hafnia alvei and CV026 was 48 mmol/L. At subinhibitory concentration, DADS could inhibit the secretion of purple pigment in CV026 regulated by quorum sensing without inhibiting growth, compared with untreated control group. At 0-12 mmol/L, with the increase of DADS concentration, the inhibition rate of DADS on biofilm and swimming ability could reach to 49.6% and 73.5%, respectively. DADS could inhibit the formation of biofilm at 12 mmol/L. It indicated that DADS could inhibit the quorum sensing activity, biofilm and swimming ability of Hafnia alvei, thus exerting antiseptic effect.
The inhibitory effect of diallyl disulfide(DADS) on biofilm and swimming ability of Hafnia alvei, a dominant putrefaction bacterium isolated from sea cucumber, was studied. The anti-quorum sensing activity of DADS was qualitatively detected using CV026 as reporter bacteria. The inhibition of DADS on biofilm and swimming activity of Hafnia alvei were analyzed by 96-microplate method, scanning electron microscopy(SEM) and agar diffusion method. The results showed that the minimum inhibitory concentration of DADS on Hafnia alvei and CV026 was 48 mmol/L. At subinhibitory concentration, DADS could inhibit the secretion of purple pigment in CV026 regulated by quorum sensing without inhibiting growth, compared with untreated control group. At 0-12 mmol/L, with the increase of DADS concentration, the inhibition rate of DADS on biofilm and swimming ability could reach to 49.6% and 73.5%, respectively. DADS could inhibit the formation of biofilm at 12 mmol/L. It indicated that DADS could inhibit the quorum sensing activity, biofilm and swimming ability of Hafnia alvei, thus exerting antiseptic effect.
引文
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[2]BASSLER B L,GREENBERG E P,STEVENS AM.Cross-species induction of luminescence in the quorum sensing bacteriumVibrio harveyi[J].Journal of Bacteriology,1997,179(12):4043-4045.
[3]HALLSTOODLEY L,COSTERTON J W,STOOD-LEY P.Bacterial biofilms:from the natural environment to infectious diseases[J].Nature Reviews Microbiology,2004,2(2):95-108.
[4]张旭东,闫超杰,孙剑,等.冷鲜牛肉中蜂房哈夫尼亚菌(Hafnia alvei)的分离与鉴定[J].微生物学杂志,2010,30(3):106-108.
[5]FUQUA C,PARSEK M R,GREENBERG E P,et al.Regulation of gene expression by cell-to-cell communication:acyl-homoserine lactone quorum sensing[J].Annual Review of Genetics,2001,35:439-468.
[6]RASMUSSEN T B,BJAMSHOLT T,SKINDER-SOE M E,et al.Screening for quorum-sensing inhibitors(QSI)by use of a novel genetic system,the QSIselector[J].Journal of Bacteriology,2005,187(5):1799-1814.
[7]刘玉平,孙宝国.含硫食用香料的合成及应用[J].中国食品添加剂,2003(6):82-84.
[8]JAKOBSEN T H,GENNIP M V,RHIPPS R K,et al.Ajoene,a sulfur-rich molecule from garlic,inhibits genes controlled by quorum sensing[J].Antimicrobial Agents and Chemotherapy,2012,56(5):2314-2325.
[9]马艳,李婷婷,崔方超,等.大菱鲆源蜂房哈夫尼亚菌群体感应现象及生物被膜调控的研究[J].现代食品科技,2016,32(8):70-76.
[10]STEPANOVICS,VUKOVI′C D,DAKI′CI,et al.A modified microtiter-plate test for quantification of staphylococcal biofilm formation[J].Journal of Microbiological Methods,2000,40(2):175-179.
[11]ADONIZIO A L,DOWNUM K,BENNETT B C,et al.Anti-quorum sensing activity of medicinal plants in southern Florida[J].Journal of Ethnopharmacology,2006,105(3):427-435.
[12]石倩,谢林香,陈立志,等.17种中草药细菌群体感应抑制剂的筛选[J].广东农业科学,2014(10):77-80.
[13]PACKIAVATHY I A,SASIKUMAR P,PANDI-AN S K,et al.Prevention of quorum-sensing-mediated biofilm development and virulence factors production in Vibrio spp.by curcumin[J].Applied Microbiology and Biotechnology,2013,97(23):10177-10187.
[14]TRUCHADO P,GIMENEZ-BASTIDA J A,LAR-ROSA M,et al.Inhibition of quorum sensing(QS)in Yersinia enterocolitica by an orange extract rich in glycosylated flavanones[J].Journal of Agricultural and Food Chemistry,2012,60(36):8885-8894.
[15]ABREU A C,BORGES A,MERGULHO F,et al.Use of phenyl isothiocyanate for biofilm prevention and control[J].International Biodeterioration and Biodegradation,2014,86:34-41.
[16]黄旭镇.大黄鱼源特定腐败菌波罗的海希瓦氏菌QS系统的鉴定及茶多酚调控研究[D].杭州:浙江工商大学,2015.