Examination of Air

Quantitative.—

Apparatus Required:

Aspirator bottle, 10 litres capacity, fitted with a delivery tube, and having its mouth closed by a perforated rubber stopper, through which passes a short length of glass tubing.

Erlenmeyer flask, 250 c.c. capacity (having a wide mouth properly plugged with wool), containing 50 c.c. sterile water.

Rubber stopper to fit the mouth of the flask, perforated with two holes, and fitted as follows:

Take a 9 cm. length of glass tubing and bend up 3 cm. at one end at right angles to the main length of tubing. Pass the long arm of the angle through one of the perforations in the stopper; plug the open end of the short arm with cotton-wool.

Take a glass funnel 5 or 6 cm. in diameter with a stem 12 cm. in length and bend the stem close up to the apex of the funnel, in a gentle curve through a quarter of a circle; pass the long stem through the other perforation in the rubber stopper.

A battery jar or a small water-bath to hold the Erlenmeyer flask when packed round with ice.

Supply of broken ice.

Rubber tubing.

Screw clamps and spring clips, for tubing.

Water steriliser.

Retort stand and clamps.

Apparatus for plating (as for enumeration of water organisms, vide page 420).

Method.—

1. Fill 10 litres of water into the aspirating bottle and attach a piece of rubber tubing with a screw clamp to the delivery tube. Open the taps fully and regulate the screw clamp, by actual experiment, so that the tube delivers 1 c.c. of water every second. The screw clamp is not touched again during the experiment.

At this rate the aspirator bottle will empty itself in just under three hours. Shut off the tap and make up the contents of the aspirator bottle to 10 litres again.

2. Sterilise the fitted rubber cork, with its funnel and tubing, by boiling in the water steriliser for ten minutes.

3. Remove the cotton-wool plug from the flask, and replace it by the rubber stopper with its fittings. Make sure that the end of the stem of the funnel is immersed in the bouillon.

4. Place the flask in a glass or metal vessel and pack it round with pounded ice. Arrange the flask with its ice casing just above the neck of the aspirator bottle.

5. Connect up the free end of the glass tube from the flask—after removing the cotton-wool plug—with the air-entry tube in the mouth of the aspirating bottle (Fig. 216).

6. Open the tap fully, and allow the water to run.

Replenish the ice from time to time if necessary.

(In emptying itself the aspirator bottle will aspirate 10 litres of air slowly through the water in the Erlenmeyer flask.)

7. When the aspiration is completed, disconnect the flask and remove it from its ice packing.

8. Liquefy three tubes of nutrient gelatine and add to them 0.5 c.c., 0.3 c.c., and 0.2 c.c., respectively, of the water from the flask, by means of a sterile graduated pipette, as in the quantitative examination of water. Pour plates.

9. Pour a second similar set of gelatine plates.

10. Incubate both sets of plates at 20° C.

11. Enumerate the colonies present in the two sets of gelatine plates after three, four, or five days and average the results from the numbers so obtained; estimate the number of micro-organisms present in 1 c.c., and then in the 50 c.c. of broth in the flask.

12. The result of air examination is usually expressed as the number of bacteria present per cubic metre (i. e., kilolitre) of air; and as the number of organisms present in the 50 c.c. water only represent those contained in 10 litres of air, the resulting figure must be multiplied by 100.

Qualitative.—

1. Proceed exactly as in the quantitative examination of air (vide supra), steps 1 to 10.

2. Pour plates of wort agar with similar quantities of the air-infected water, and incubate at 37° C.

3. Pour plates of nutrient agar with similar quantities of the water and incubate at 37° C.

4. Pour similar plates of wort gelatine and incubate at 20° C.

5. Pick off the individual colonies that appear in the several plates, subcultivate them on the various media, and identify them.