【摘要】 依据逆向调整原则,设计了4套诱导乳酸杆菌进入活的非可培养(VBNC)状态的方案,同时应用LIVE/DEAD细胞染色试剂,对进入该状态的细菌进行了荧光观察,并尝试用Tween-80对VBNC状态细菌进行了复苏。结果显示,MRS液体培养基,pH 2.0~3.0,4℃,兼性厌氧培养和不含半胱氨酸的MRS液体培养基,pH 2.0~3.0,4℃,兼性厌氧培养均可使3株试验菌体在短期内进入VBNC状态,并且能较长时间维持这一状态;3株细菌菌体均发生萎缩、弯曲,且彼此相连,原本单在的杆菌变成了链杆菌;随着诱导时间的延长,菌体的交联程度明显增加,出现众多红绿交替的长链杆菌;Tween-80可使VBNC状态细菌恢复为可培养状态并实现复苏,而复苏后的细菌重新具备原菌的形态和生化特征。表明,乳酸杆菌具有VBNC状态,并且在此状态下呈现与正常菌体不同的形态特征。更多还原

【Abstract】 Four induction protocols were designed for Lactobacillus lactis entering into viable but non-culturable(VBNC) state according to reverse adjustment protocol.Fluorescent images from VBNC L.lactis were observed with Live/Dead Bacterial Viability Kit.The VBNC bacterium resuscitation was conducted with Tween-80.Both protocol 2 in which L.lactis strains were cultured facultatively and anaerobicly with MRS from pH 2.0 to 3.0 at 4 ℃ and protocol 4 in which L.lactis strains were cultured facultatively and anaerobicly with MRS without cysteine from pH 2.0 to 3.0 at 4 ℃ could induce L.lactis strains to become VBNC state in a short time and maintain the state in a long time.Fluorescent observation showed that the cells of 3 strains were all atrophy,bending,connecting with each other,and altering original bacillus into streptobacillus.Connecting with each other was obviously increasing with the induction time,many streptobacillus showed red or green.VBNC L.lactis could be resuscitated to normal morphous and biochemical characteristics with Tween-80.The results indicated that L.lactis could become into VBNC state with different morphous.更多还原