1、快速記數法,記數PetrifilmATP鑑定API 20E,Petrifilm,製作公司: 3M將一培養皿 (petri dish) 製作在一薄膜中 (film),故稱為 petrifilm 已成為工業界標準操作含有水溶膠, 營養素,指示劑 Size: 10 cm 7.5 cm (about 20 cm2).1 cm 1 cm yellow grid is printed on the back of the plate to assist enumeration. 塑膠 “spreader” 塗抹片.,與傳統方法比較,不用準備培養基減少廢棄物減少體積增加測試量 (0.1 vs. 1 mL)有
2、各式產品可供不同的目標菌,種類,總菌數 (total aerobic plate count, APC, TPC)通用性培養基 general mediumTSA, TSB, Nutrient agar, BHI 大腸桿菌群/大腸桿菌 (coliform/E. coli)金黃色葡萄球菌 (Staphylcoccus aureus)選擇性培養基 selective medium只有目標菌生長特殊型態與顏色,Aerobic Count Plate,洋菜膠透氣膜細菌生長Produce acid, lower pH, indicator turn red,大腸桿菌群/大腸桿菌 (coliform/E.
3、 coli),大腸桿菌群Gram 陰性、非胞子、能將乳糖發酵產生氫與CO2Escherichia coli, Citrobacter freundii, Enterobacter aerogenes, Enterobacter cloacae, Klebsiella penumoniae主要在腸道內,可在自然界中生長糞便大腸桿菌群 (fecal coliform)44.5 亦可在自然界中生長,需要較多有機質,主要在腸道內大腸桿菌Escherichia coli (E. coli),Petrifilm E. coli/Coliform Count (EC) plates contain Viol
4、et Red Bile (VRB) 紫紅膽鹽、膠、指示劑、葡萄糖醛 酸酶 ( -glucuronidase )紫紅膽鹽限制G+ 菌生長大腸桿菌群紅+氣泡大腸桿菌藍+氣泡 葡萄糖醛酸酶 ( -glucuronidase ),大腸桿菌 (Escherichia coli),一般型大多無害指標菌腹瀉型感染型出血型中間形,Materials and methods,Bacteria:Escherichia coli ATCC 11775 Enterobacter aerogenesPetrifilmAPCcoliformcoliform/E. coli,Bacteria mixture (from a
5、ll two bacterial species)One tube/benchDilution dilute to -8 (8 tubes/group)APCAdd one mL of -7 and -8Each dilution has two films (total 4 films)ColiformAdd one mL of -8 Each dilution has one film (total 1 films)E. coli/coliformAdd one mL of -8 Each dilution has one film (total 1 films),APCRed coloniesColiform Red with air bubbleE. coli Blue with air bubble,取 1 ml稀釋液滴於Petrifilm上,APC: 35, 48 hourColiform/E. coli: 24 h incubationStaphylcoccus: 24 hour,APC,E. coli/coliform,