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GB 4789.5-2012: National food safety standard -- Microbiological examination of food hygiene -- Examination of shigella Delivery: 9 seconds. True-PDF full-copy in English & invoice will be downloaded + auto-delivered via email. See step-by-step procedure Status: Valid GB 4789.5: Historical versions
Similar standardsGB 4789.5-2012: National food safety standard -- Microbiological examination of food hygiene -- Examination of shigella---This is an excerpt. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.), auto-downloaded/delivered in 9 seconds, can be purchased online: https://www.ChineseStandard.net/PDF.aspx/GB4789.5-2012 GB NATIONAL STANDARD National Food Safety Standard Microbiological Examination of Food Hygiene - Examination of Shigella Issued on. MAY 17, 2012 Implemented on. JULY 17, 2012 Issued by. Ministry of Health of the People’s Republic of China Table of ContentsForeword... 7 1 Scope... 8 2 Devices and Materials... 8 3 Culture Medium and Reagents... 8 4 Examination Procedures... 9 5 Operational Procedures... 10 Appendix A... 15ForewordThis Standard replaces GB/T 4789.5-2003 "Microbiological Examination of Food Hygiene - Examination of Shigella". Compared with GB/T 4789.5-2003, main changes in this Standard are as follows. - The standard name was modified; - Culture mediums and reagents were modified; - Enrichment part, biochemical test and additional biochemical test part in the operational procedures were modified; - Table 2 was modified; - Table 4 was modified. National Food Safety Standard Microbiological Examination of Food Hygiene - Examination of Shigella1 ScopeThis Standard specifies the methods to examine the Shigella in foods. This Standard is applicable to the examination of Shigella in foods.2 Devices and MaterialsIn addition to the conventional sterilization and cultivation devices in the microbiological laboratory, other devices and materials needed for the examination are as follows.3 Culture Medium and Reagents3.1 Enriched Shigella broth - novobiocin. see A.1 in Appendix A. 3.2 MAC agar. see A.2 in Appendix A. 3.5 TSI agar. see A.4 in Appendix A. 3.6 Nutrient agar slant. see A.5 in Appendix A. 3.7 Semi-solid agar. see A.6 in Appendix A. 3.8 Ammonium dextrose medium. see A.7 in Appendix A. 3.9 Urea agar. see A.8 in Appendix A. 3.15 Peptone water, and indole reagent. see A.14 in Appendix A. 3.16 Shigella diagnostic serum. 3.17 Biochemical identification reagent kit.4 Examination ProceduresSee Figure 1 for the Shigella examination procedures. Examined Sample5 Operational Procedures5.1 Enrichment Take 25g (mL) of examined sample by sterile operation, add it into a homogenizing cup containing 225mL of sterile enriched Shigella broth, homogenize it with a spinning-blade homogenizer at 8000r/min~10000r/min; 5.2 Separation Take the enriched Shigella enrichment liquid, and inoculate it on XLD agar plate and MAC agar plate or Shigella chromogenic medium plate respectively. Cultivate it at 36ºC±1ºC for 20h~24h, observe the morphology of colonies growing on each plate. The diameter of single Shigella sonnei colony shall be larger than other Shigella's. 5.3 Preliminary biochemical test 5.3.1 Pick up more than 2 typical or questionable bacterial colonies from the selective agar plate, inoculate them on a tube of TSI, semi-solid and nutrient agar slant respectively, cultivate them at 36ºC±1ºC for 20h~24h, observe the results respectively. 5.4 Biochemical test and additional biochemical test 5.4.1 Biochemical test Use the lawn which is growing on the nutrient agar slant and is cultivated in 5.3.1, to perform biochemical test, that is the decomposition test of β-galactosidase, urea, lysine decarboxylase, ornithine decarboxylase, salicin and esculin. 5.5 Serological identification 5.5.1 Preparation of antigen Shigella has no power, therefore it has no flagellar antigen. The main antigen in the Shigella is thallus (O) antigen. Thallus (O) antigen may be subdivided to type and group specific antigen. 5.5.2 Agglutination reaction Draw out 2 areas of 1cm×2cm on the slide, pick up a loop of bacteria to be tested, place 1/2 loop on the upper part of each area respectively; add a drop of antiserum on the lower part of one area, add a drop of normal saline on the lower part of the other area for comparison. Grind the colonies in the two areas to emulsion respectively with a sterile inoculating loopAppendix ACulture Medium and Reagents A.1 Enriched Shigella broth - novobiocin (Shigella broth) A.1.1 Enriched Shigella broth A.1.2 Novobiocin solution A.1.2.1 Ingredients Novobiocin 25.0mg Distilled water 1000.0mL A.1.2.2 Preparation Dissolve the novobiocin into the distilled water, remove the bacteria with 0.22μm membrane filter; if the solution is not used immediately, it may be stored at 2ºC~8ºC for a month. A.1.3 5mL of novobiocin solution (A.1.2) is added in per 225mL of enriched Shigella broth (A.1.1) and mixed uniformly before use. A.2 MAC agar A.4 ISI agar A.5 Nutrient agar slant A.5.2 Preparation Except for agar, dissolve the rest ingredients into distilled water, add about 2mL of 15% sodium hydroxide solution, cool the solution to around 25ºC, calibrate pH to 7.0±0.2.Add agar, boil the solution to dissolve the agar. Dispense the solution into small test tubes, each tube has about 3mL. Sterilize the solution at 121ºC for 15 min, make it into slant. A.6 Semi-solid agar A.6.1 Ingredients ......Source: Above contents are excerpted from the full-copy PDF -- translated/reviewed by: www.ChineseStandard.net / Wayne Zheng et al. 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