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E. coli group test procedure report – europuppyblog

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  1. Elastic flora refers to a group of fermented lactose, acidic acid (the original color of the medium is red, if acidic acid becomes yellow), Gram -negatives who produce gas, oxygen, and imponent anaerobic are negative. Bacillus -free. Food detects the colorectal flora, which indicates that the food was contaminated by feces. Results The report is the nearest number (MPN)
    device and materials of the colon flora per 100ml (g) sample:
    1. Temperature box: 36 ± 1 degree
    2. Refrigerator: 0: 0 -4 degrees
    3. constant temperature water bath: 44.5 ± 0.5 degrees
    4. Teacher
    5. Microscope
    6. Modifier or milk benzene
    7. Ping dish: 90mm
    8. IVF
    9. Plant
    10. Wide mouth bottle or triangular bottle: capacity is 500ml
    11. Glass beads: diameter is about 5mm
    12. Alcohol lamps
    13. IVIT
    Cultivation bases and reagents:
    1. Lactose gallbladder salt fermentation pipe
    2. Yonghongmei blue agar tablets
    . 3. 3. Lactan fermentation tube
    4. EC broth
    5. Phosphoric acid buffer diluted liquid
    . physiological saline
    7. Gram -staining solution
    Operation steps:
    7.1). Dilute the sample
    7.1.1 Place the sample of 25ml (or g) with a sterile operation in 225ml sterilized physiological saline or other diluted glass bottle (the appropriate quantity in the bottle is placed in the bottle Glass beads) or sterilized breast phenyl phenyl, through full shaking or grinding into a uniform diluted solution of 1:10. It is best to use a homogeneous device for solid testing, and treat mining min at a speed of 8000-10000R/min to make a uniform diluted solution of 1:10.
    7.1.2 Use a 1ml sterilization straw to absorb 1 ml of diluted solution, inject a test tube containing 9ml sterilized physiological saline or other diluted liquids, shake the tube mixing, and make 1: 100 dilution liquid r r r r r r r r r
    7.1.3 Take another 1ml sterilization straw. Make 10 times the diluted liquid in turn according to the above operation.
    7.1.4 According to the requirements of food hygiene standards or estimates of the pollution of the test, choose three dilution degrees and each dilution degree is vaccinated with 3 tubes.
    7.2 Lactan fermentation test
    Mainer the sample to be checked in the lactose gallbladder fermentation pipe, the amount of inoculation is more than 1ml, and the two lactose gallbladder salt fermentation tubes are used. Each dilution is inoculated and 36 ± 1 degrees the temperature box, and 24 ± 2 hours. If all lactose gallbladder salt fermentation tubes do not produce qi, you can report it as the colonic flora negative. According to the following procedures.
    7.3 Separation Cultivation
    The fermentation tube of Qi-producing gas is inachened on Yonghongmei Blue Graphon Flate, set up 36 ± 1 degree temperature box, cultivate 18-24 hours, then take it out to observe the lycomeda form of the lycomeda form. , And do Gram -colored dyeing and experimental tests.
    7.4 Confirmation of the test
    On the above tablet, pick up the suspicious colon flora 1-2 for Gram-dyeing, and inoculate the lactose fermentation tube at the same time. In 2 hours, observing gas -producing gas -producing gas -free aphid -free aphids, and the Gram -gyan dyeing can be reported as positive of colonbaccinal groups.
    7.5 Report
    In the confirmation of the number of colonous pipes for colonobacteria, check the MPN retrieval table, and report the MPN value of each 10ml (g) colon flora.
    8 The dung colon flora
    8.1 Cultivation of all gas -producing lactose gallbladder fermentation tubes (see 7.2) in the EC broth tube, set up 44.5 ± 0.2 degrees of water bath boxes (The water surface in the water bath should be higher than the EC broth surface), and it is cultivated for 24 ± 2 hours. After cultivation, if all EC broth tubes do not produce qi, they can be reported as negative; All the gas-producing EC broth is inaged on the Yonghongmei Blue Graphon Flate, and placed 36 ± 1 degree to cultivate for 18-24 hours. Those who have typical colonies on the tablet are confirmed to be positive for the fecal colonobacterus.
    8.2 Results report
    In the confirmation of the positive tube of the dung colon flora, check the MPN retrieval form, and report to the MPN value of each 100ml (g) the mpn of the dung colon flora

  2. Detective process of dung colon flora in cosmetics detection projects:
    10g/ml sample 90ml sterilized physiological saline → 10ml 10ml double lactose bile (including neutral agent) medium → fecal colonobacteria 44.5 ° C, 48H Cultivation
    Note: In cosmetics detection items, if the sample contains oil -grease ingredients, such as essential oils, liquid paraffin, vomiting vomiting 80, and physiological saline should be added in front of the sample, so that the sample can be evenly dispersed and open. Come.
    What kind of microorganisms are the dung colorectal flora? Let ’s introduce it in detail:
    Elac bacteria: colonobacteria is not named for bacterial classification, but the term of the field of sanitary bacteria. It does not represent a certain or certain bacteria. A set of characteristics of bacteria related to feces pollution, these bacteria are not completely consistent in biochemical and serotology.
    Definition (GB2010): Under certain cultivation conditions, it can ferment lactose, acidic gas, oxygen, and concentrated anaerobic Gram -negative Bacillus.
    This Eschaous population: a type of colon flora, also known as thermal large -embodiments, cultivated temperature: 44.5 ± 0.5 ° C. The fecal colonic flora is a microbial project that must be tested in cosmetics testing projects.
    Edimum Erus bacteria: ((. Coli) is usually called E. coli. According to different biological characteristics, the pathogenic E. coli is divided into 5 categories:
    ,
    The bowel toxic E. coli (ETEC),
    EAEC).
    The test service you may be interested in:

  3. Because the E. coli group refers to a group of bacteria related to stool pollution with certain characteristics, that is, the Gram -negative germiosilus of Gram -based germinated germinated Bacillus of Gram -based germinated germinated in 37 ° C. Therefore, the detection of the colorectal flora is generally performed in accordance with its definition.
    The testing method of import and export food colorectal flora adopted in China mainly includes national standards and industry standards formulated by the former State Commercial Inspection Bureau. The two standard methods are slightly different in the detection program.
    (1) National standards: National standards adopt three steps, namely: lactose fermentation test, separation training, and confirmation test.
    The lactose fermentation test: After the sample is diluted, select three dilution degrees, and each dilution degree to inoculate tri -tube lactose gallbladder fermentation tubes. Cultivate 48 ± 2h at 36 ± 1 ℃ to observe whether it is produced.
    Sepan cultivation: Turn the cultivation of gas production tubes on Yonghongmei Blue Graphite tablet, and cultivate 18-24h at 36 ± 1 ℃ to observe the formation of the colonies.
    Crofin test: Pick the suspected colonies on the tablet and observe Gram -dyeing. At the same time, the lactose fermentation tube is 36 ± 1 ℃ to cultivate 24 ± 2h to observe the gas production situation.
    Report: According to the number of positive pipes of E. coli, check the MPN table and report the MPN value of the Escharian flora per 1ml (g). For specific operations, see
    gb4789.3-2010 “National Standard Food Sanitary Microbiology Test of the People’s Republic of China
    Thenexal flora measurement”
    The industry standards formulated by the bureau, equivalent uses the US FDA standard method to detect E. coli in exported foods. This method adopts two steps:
    G speculation test: After the sample is diluted, select three dilution degrees, and each dilution degree to inoculate tri -tube LST soup. Cultivate 48 ± 2h at 36 ± 1 ℃ to observe whether it is produced.
    Keng test: Incubate the gas -producing green lactose gallbladder salt (BGLB) soup tube, cultivate 48 ± 2h in 36 ± 1 ° C to observe whether the gas is produced. BGLB is positive. Check the MPN table and report the mpn value of the colonic flora in each ml (g) sample.
    The specific operation see SN0169-92

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