2492 Journal of Applied Microscopy 



[ NEWS AND NOTES. j 



Culture media in test-tubes may be solidified either in an upright or a slant- 

 ing position. The former is the one most useful for stab cultures, but the slant- 

 ing position offers a larger and easier inoculating area and can be used for stab 

 cultures if required. To inoculate a tube it should be held between the first 

 finger and thumb of the left hand, with the slanting surface of the medium point- 

 ing upwards, and by keeping the tube as nearly horizontal as possible any germs 

 in the air (which have a weight of their own) are prevented from falling into it. 

 The infective matter, or second tube, should be held between the first and 

 second fingers of the same hand, and, taking the needle high up the handle by 

 the thumb and first two fingers of the right hand, sterilize it by placing in the 

 flame till red hot, then with the little finger and palm, or with the backs of the 

 ring and little fingers also of the right hand, remove the cotton-wool plug from 

 the infective tube by a twisting movement. This screwing motion removes the 

 cotton-wool fibres from the mouth of the tube more thoroughly than would be 

 the case if the plug was drawn straight out. And now, as quickly and cleanly 

 as possible, remove from the infective tube a very little of the specimen on the 

 point of the needle. Plug the tube ; remove the wool from the fresh one, and 

 inoculate the new medium by making one, two, or three streaks the full length 

 of the culture surface, and avoid touching the side of the tube. Withdraw the 

 needle ; pass the mouth of the test-tube two or three times through the flame, do 

 the same with the plug end of the cotton-wool, and insert it firmly in the tube. 

 Sterilize the needle before laying down. Label the tube with date and hour of 

 inoculating, and the source of the infective material, taking care not to wet the 

 label with the tongue, as we are dealing with infective matter. Place in the 

 incubator. In the case of stab cultures, the infective material is planted by 

 thrusting the straight needle into the substance of the nutrient medium for its 

 full length.- — Jour. State Med. 



An apparatus for making anaerobic cultivations in fluid media consists of a 

 round-bottomed flask or stout bottle (A) fitted with a single-bored stopper, 

 through which passes a right-angled bend of manometer tube (B). This is con- 

 nected by thick-walled rubber tubing with another bend of glass tube (C), which 

 passes through a cotton-wool plug into a test-tube or small flask (D) ; the rubber 

 connection carries a screw or spring clip. The apparatus having been sterilized 

 by moist heat, the bottle is nearly filled with the medium, about 15 c. c. of which 

 are placed in the tube D ; the whole is then sterilized for half an hour on two 

 successive days in the steamer, the clip being left open, and the tube C being 

 drawn up above the fluid in D. On the third day the bottle is placed in a cal- 

 cium chloride bath (immersed up to its neck), which is maintained at a tempera- 

 ture of about 120° C, until all the air is driven out (five minutes will suffice to 

 displace 50 c. c. of air) ; the tube C is then lowered into the medium, and the 

 clip fastened. To inoculate the medium in the bottle a fluid culture or broth 

 emulsion must be used ; the tube C is removed from the test-tube and dipped 

 into the culture ; on opening the clip the infected fluid in the tube will be sucked 

 into the bottle, and when a sufficient amount has been transferred the clip is 

 again shut. The condition in the bottle is one of complete anaerobiosis, and the 

 partial vacuum does not exercise any ill-effect upon the growth of the bacteria. 

 — Jour. State Med. 



