百事微生物检测方法.docVIP

百事：微生物检测方法 1. 目的 PURPOSE 概述非碳酸饮料产品的耐酸细菌、酵母菌和霉菌的分析程序。To outline the procedure for analysis of NCB products for Aciduric Bacteria, Yeast and Mould. 2. 范围 SCOPE 本程序适用于需要精确测定存在的耐酸细菌、酵母菌和霉菌的非碳酸饮料产品。这些有机物有可能导致饮料的腐败。采用浇灌平板法对这些参数进行分析，酵母菌和霉菌用的是PDA，耐酸细菌用的是OSA。The procedure is applicable to NCB products where an accurate enumeration of the Aciduric Bacteria, Yeast and Mould present is required. These organisms may potentially cause spoilage of beverages. Analysis for these parameters is conducted using a pour plate methods using PDA for Yeast and Mould and OSA for Aciduric Bacteria. 3. 程序 PROCEDURE 3.1 设备 Equipment - 无菌移液管 – 10ml容量Sterile Pipettes – 10ml Capacity - 无菌的皮氏培养皿– 150mmSterile Petri Dishes – 150mm - 设定为30°C +/- 1°C的恒温箱Incubator set at 30°C +/- 1°C - 设定为25°C +/- 1°C的恒温箱Incubator set at 25°C +/- 1°C - 恒温水浴，45 +/- 1°CConstant Temperature Waterbath at 45 +/- 1°C - 高压蒸汽灭菌器Autoclave - 微波Microwave - 纸巾Tissues - 0.2 μm过滤器0.2 μm Filter - 注射器(5ml或10ml)Syringe (5ml or 10ml) 3.2 试剂 Reagents - 橘汁琼脂（Oxoid或同类产品）Orange Serum Agar (Oxoid or Equivalent) - 马铃薯右旋糖琼脂（Oxoid或同类产品）Potato Dextrose Agar (Oxoid or Equivalent) - 去离子水Deionised Water - 乙醇/异丙醇~70%Ethyl/Iso-propyl Alcohol ~70% 3.3 标准 Standards 不适用N/A 3.4 试验的准备 Test Preparation 3.4.1 工作台的准备 Preparation of Workstation 层流装置的准备（任选）Laminar Flow Preparation (Optional) ? 清洁装置并喷上70%的异丙醇或普通酒精，并用纸巾擦干；或让其蒸发。保证载片是关闭的，同时打开紫外灯。务必保证下列条件：Clean the cabinet and spray with 70% isopropyl or ethyl alcohol and dry with tissues or allow to evaporate. Ensure the sliding glass is closed and turn on the UV lamp. Always ensure the following: - 使用紫外线时滑动窗要保持关闭。Keep the sliding window closed when UV is in use. - 打开滑动窗之前要关闭紫外灯。Turn off the UV light before open the sliding window. - 样品、培养基或者你本人在任何时候都不得接触紫外线。Do not expose samples, media or yourself to UV light at any time. ? 试验之前至少15分钟就要接通装置的鼓风机，以便清除装置中的空气并代之以经过过滤的空气。Turn ON the cabinet blower at least 15 minutes before testing to allow the cabinet purge the air inside the c