Studies on Agar Culture Media for Mycobacterium Tuberculosis VI. Modification of agar culture media by using stored whole blood. 2.
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The authors have been trying to improve twokinds of agar culture medium for Myco. tuberculosis, that is, the medium for inoculation of 0.1 cc of specimen treated with 4% NaOHand that for 0.1 cc of neutralized specimen.In the course of this study, some strains of Myco. tuberculosis were found to grow morepoorly and contaminations by other organismswere experienced more frequently on these mediathan on the egg culture medium.<BR>For the purpose of preparing a medium whichis more suitable for the growth of the Myco.tuberculosis, less frequently contaminated byother organisms, and has more simple constituentsthan the conventional media, the followingpointswere investigated: 1) a favorableamount of potassium phosphate, monobasic, and sodium phosphate, dibasic which were tobe added in the medium; 2) whether or not anadditional magnesium sulfate, sodium citrate, asparagine or sodium glutamate etc. were requisitefor the growth of Myco. tuberculosis;and 3) an appropriate amount of malachitegreen or penicillin to inhibit the growth ofcontaminants. Consequently a modified mediumwas prepared, having the following composition.<BR>1) A medium for inoculation of 0.1 cc of4 % NaOH treated specimen<BR>KH<SUB>2</SUB>PO<SUB>4</SUB>…0.5g<BR>Na<SUB>2</SUB>HPO<SUB>4</SUB>·12H<SUB>2</SUB>O …0.3g<BR>Sodium glutamate…0.3g<BR>Soluble starch …2.0g<BR>Glycerine …1.5cc<BR>Purified agar …2.0g<BR>0.1% malachite green …3.0cc<BR>(or Penicillin …100u/cc)<BR>Distilled water …100.0cc<BR>These constituents were dissolved, sterilizedin an autoclave and cooled to 55°C to 60°C, to which stored whole blood was added andmixed well to make its final concentration 10%.Five cubic centimeters of the medium wasdispensed into each test tube to make a slant, whose pH would be around 6.25 and becomearound 7.04 after the addition of 0.1 cc of 4%NaOH.<BR>2) A medium for inoculation of 0.1 cc ofneutralized specimen<BR>The amount of KH<SUB>2</SUB>PO<SUB>4</SUB> in the medium 1) wasreduced from 0.5g to 0.1g.<BR>That of other constituents were the same asabove. pH of this medium was around 6.63.<BR>More colonies were noted on these mediathan on a conventional stored whole blood agarmedium, 3 % and 1% Ogawa egg media.
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