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Total aerobic microbial count (Multiple -tube method:(
1-Suspend 10.0 g(solid) or 10.0 ml(liquid) of the specimen influid soybean casein digest medium
(in PH7.2 phosphate buffer) or buffered sodium colored-peptone solution PH7.0to make 100 ml.It will be 1:10 dilution (1/10)
. It is calledpretreatment material
Add the specimen to medium not more than 1 hour after preparing the appropriatedilutions for inoculation.2-Prepare a series of 12 tubes each containing 9-10 ml of soybean-casein digest medium.
3-Divide these 12 tubes in four sets of three tubes each.
4-To each of the first three tubes add 1 ml of 1:10 dilution described in no 1.
5-To the next three tubes, add 1 ml of a 1:100 dilution described in no 4.
6-To the next three tubes, add 1 ml of 1:1000 dilution described in no 5.
7-To the last three tubes, add 1 ml of the diluent (as control(
8-Incubate the tubes at 30-35°C for at least 5 days N.B; 1- No microbial growth should appear in the last three tubes.
2-If the reading is difficult, prepare a subculture in a liquid and evaluate the result after further period of incubation.9-Determine the most probable number of M.O per gm or ml of material using the next table.
If, for the first column, the number of tubes showing microbial growth is two or less, themost probable number of M.O per gm or per ml is less than 100. References;
1-Herbal Medicines Manufacturing Guidelines, Ministry of Health Egypt.http://www.mohp.gov.eg/Sec/Statistics/PhBooks.asp?x=32-Quality Control Methods for Medicinal Plant Materials, WHO Geneva.
3-WHO Guidelines on GMP for Herbal Medicines. ISBN 978 9241 547161.
4-laboratory practice in tropical countries Monica Cheesbrough 2000, ISBN 977-05-2050-0
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