医学微生物学资料：细菌实习-2 2

1、CAMS/PUMC细菌学实习-II多途径细菌采样培养及初步鉴定CAMS/PUMC实验材料咽拭子 1支/人大号普通培养皿 2块/人小号羊血培养皿 2块/人10ml无菌肉汤管 1支/人载玻片 1片/人 A、B菌 3套/桌NS 1份/人滤纸 1张/人无菌棉棒CAMS/PUMC实验目的了解各种培养基的制备掌握机体常见部位及环境中的细菌采样、培养及初步鉴定掌握革兰氏染色方法及结果判定CAMS/PUMC实验内容机体不同部位的细菌采样、培养咽部正常菌群的采样、培养未消毒手的细菌采样培养CAMS/PUMC实验内容环境中细菌采样培养常用生活用品的细菌采样培养手机流通纸币抗菌洗浴用品抑菌功效的检测CAMS/PUM

2、C紫外线穿透能力试验CAMS/PUMC实验内容A、B菌鉴定（Gram stain）报告内容染色结果细菌形态CAMS/PUMC细菌涂片制备程序取一干净玻片，火焰上方缓缓通过三次去油蜡笔画线将玻片分割为A、B两区域接种环取NS分别滴入A、B两区域各一滴分别取少许A、B菌 ，与NS充分混匀成菌液，风干后形成菌膜固定（火焰上方缓缓通过三次）革兰氏染色滤纸吸干水分，油镜观察CAMS/PUMC Heat fixingCAMS/PUMCSmear preparation CAMS/PUMCCAMS/PUMCcrystal violet stainCAMS/PUMCwash with water CAMS/P

3、UMCStain with grams iodine solution CAMS/PUMCDecolorize by adding 95% alcohol CAMS/PUMC Counterstain with Safranin CAMS/PUMCCAMS/PUMCG+菌G-菌CAMS/PUMCCAMS/PUMCCAMS/PUMCCAMS/PUMC细菌接种法平行划线法分区划线法密集接种法CAMS/PUMC平行划线法CAMS/PUMC分区划线法CAMS/PUMCbackCAMS/PUMCCAMS/PUMC培养基（Culture medium）Substances used for artific

4、ial cultivation of bacteria. CAMS/PUMCNutrient requirements of bacterial growth water Carbon source Nitrogen source Minerals Growth factors organic compounds such as amino acids, purines嘌呤, pyrimidines嘧啶, and vitamins that a cell must have for growth but cannot synthesize itself. CAMS/PUMCClassifica

5、tion of culture mediumAccording to basic ingredients:Minimal essential growth medium基础培养基enriched medium营养培养基selective medium选择培养基differential medium鉴别培养基CAMS/PUMCAccording to physical conditionliquid medium液体培养基solid medium固体培养基 1.52.5% of agar semisolid medium半固体培养基0.30.5% of agar CAMS/PUMC Phenom

6、ena of bacterial growth细菌在培养基中的生长情况In liquid medium： surface growth pellicle表面生长, uniformly turbid均匀混浊生长, sediment in bottom沉淀生长 CAMS/PUMC菌膜菌沉淀均匀浑浊对照CAMS/PUMCCAMS/PUMC抗菌用品抑菌功效试验步骤将同一人双前臂暴露自来水冲洗数秒分别用舒肤佳及夏士莲香皂，自上而下涂抹前臂三次自来水冲洗数秒将琼脂棒贴在双前臂皮肤表面5分钟放入无菌培养皿中37 培养24小时通过菌落数的观察判断抑菌效果咽部正常菌群的采样、培养咽拭子擦拭咽部，平行划线法接种于羊血平皿CAMS/PUMCCAMS/PUMCCAMS/PUMC未消毒手的细菌采样培养将10 ml液体培养基加入无菌手套内充分洗手混匀后吸出用棉棒进行密集接种CAMS/PUMC密集接种用无菌棉棒儿蘸取菌液将多余菌液在液面上方管壁内轻轻旋转挤出，涂布在琼脂平板整个平面三次，每次旋转60度，最后沿平板内缘涂抹一周，以取得均匀接种紫外线穿透能力试验密集接种法接种细菌将不同形状的纸片贴至培养基