1、CAMS/PUMC細菌學實習-II多途徑細菌采樣培養及初步鑒定CAMS/PUMC實驗材料咽拭子 1支/人大號普通培養皿 2塊/人小號羊血培養皿 2塊/人10ml無菌肉湯管 1支/人載玻片 1片/人 A、B菌 3套/桌NS 1份/人濾紙 1張/人無菌棉棒CAMS/PUMC實驗目的了解各種培養基的制備掌握機體常見部位及環境中的細菌采樣、培養及初步鑒定掌握革蘭氏染色方法及結果判定CAMS/PUMC實驗內容機體不同部位的細菌采樣、培養咽部正常菌群的采樣、培養未消毒手的細菌采樣培養CAMS/PUMC實驗內容環境中細菌采樣培養常用生活用品的細菌采樣培養手機流通紙幣抗菌洗浴用品抑菌功效的檢測CAMS/PUM

2、C紫外線穿透能力試驗CAMS/PUMC實驗內容A、B菌鑒定（Gram stain）報告內容染色結果細菌形態CAMS/PUMC細菌涂片制備程序取一干凈玻片，火焰上方緩緩通過三次去油蠟筆畫線將玻片分割為A、B兩區域接種環取NS分別滴入A、B兩區域各一滴分別取少許A、B菌 ，與NS充分混勻成菌液，風干后形成菌膜固定（火焰上方緩緩通過三次）革蘭氏染色濾紙吸干水分，油鏡觀察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細菌接種法平行劃線法分區劃線法密集接種法CAMS/PUMC平行劃線法CAMS/PUMC分區劃線法CAMS/PUMCbackCAMS/PUMCCAMS/PUMC培養基（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基礎培養基enriched medium營養培養基selective medium選擇培養基differential medium鑒別培養基CAMS/PUMCAccording to physical conditionliquid medium液體培養基solid medium固體培養基 1.52.5% of agar semisolid medium半固體培養基0.30.5% of agar CAMS/PUMC Phenom

6、ena of bacterial growth細菌在培養基中的生長情況In liquid medium： surface growth pellicle表面生長, uniformly turbid均勻混濁生長, sediment in bottom沉淀生長 CAMS/PUMC菌膜菌沉淀均勻渾濁對照CAMS/PUMCCAMS/PUMC抗菌用品抑菌功效試驗步驟將同一人雙前臂暴露自來水沖洗數秒分別用舒膚佳及夏士蓮香皂，自上而下涂抹前臂三次自來水沖洗數秒將瓊脂棒貼在雙前臂皮膚表面5分鐘放入無菌培養皿中37 培養24小時通過菌落數的觀察判斷抑菌效果咽部正常菌群的采樣、培養咽拭子擦拭咽部，平行劃線法接種于羊血平皿CAMS/PUMCCAMS/PUMCCAMS/PUMC未消毒手的細菌采樣培養將10 ml液體培養基加入無菌手套內充分洗手混勻后吸出用棉棒進行密集接種CAMS/PUMC密集接種用無菌棉棒兒蘸取菌液將多余菌液在液面上方管壁內輕輕旋轉擠出，涂布在瓊脂平板整個平面三次，每次旋轉60度，最后沿平板內緣涂抹一周，以取得均勻接種紫外線穿透能力試驗密集接種法接種細菌將不同形狀的紙片貼至培養基