ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 
101 
Take a moderately large tin, suck as a biscuit tin, and a wooden box 
of sufficient size to give 3 or 4 inches clear space in every direction when 
the tin is placed within it. Put a layer of sawdust 3 or 4 inches deep at 
the bottom ; stand -the tin upon this, and fill the surrounding space with 
sawdust also. The various organs, after being removed with as much 
cleanliness as possible, are each wrapped in a clean cloth wetted (not 
wringing wet) with 1-500 Hg01 2 , and further wrapped and sealed each 
separately in a piece' of gutta-percha tissue (most easily done with a 
trace of chloroform) ; they are then put into one or more watertight tins 
(those 1/4 or 1/2 lb. tins in which some tobacco is sent out are admir- 
able ; they may be more securely closed by means of a paraffin candle 
and a warm poker). The tobacco tins are placed within the larger tin, 
together with pounded ice and common salt, in the proportion of 3 lbs. 
of ice to 1 lb. of salt. The lid is placed on the larger tin, and the 
wooden outer box filled up with sawdust. An apparatus which holds the 
above quantities of ice and salt will keep about 1/2 pint hard frozen for 
more than twenty-four hours. There can be no difficulty in obtaining 
these materials at a moment’s notice. The maintenance of objects at a 
low temperature when bacteriological examination cannot be carried out 
completely on the spot, is essential for satisfactory results to accrue. 
Demonstrating the Coccidia of Cuttle-fish.* — The specimens used 
by M. M. Siedlecki were obtained from the Gulf of Naples, and all the 
animals examined were found to be more or less infected. The parasites, 
localised in the digestive apparatus, are recognisable by the naked eye as 
white opaque spots in the posterior part of the intestine. Observations 
were made on fresh and also on fixed and stained specimens. 
Fresh specimens were placed on cover- glasses the corners of which 
had been slightly melted in the flame of a Bunsen’s burner. In this way 
the cover was supported on four footlets, and crushing of the prepara- 
tions avoided. The specimen was immersed in sea-water or in intestinal 
juice, was broken up as little as possible, and then placed on a slide for 
examination. 
Stained preparations were made from teased-out pieces and from 
sections, the former giving the best results. A piece of intestinal mu- 
cosa and submucosa, from 5 to 10 mm. square, was placed on a cover- 
glass, teased out in sea- water or in intestinal juice, and carefully spread 
over the whole surface of the cover. The cover was then placed film 
side down on the surface of the fixative solution. In this way the layer 
. is at once fixed, and also caused to adhere to the cover. If the fluid (sea- 
water or juice) be in excess, the adhesion of the film will be imperfect, 
and some of the preparation will drop off ; while if the film be allowed to 
dry before being fixed, the structure of the Coccidia is profoundly altered 
or destroyed. 
The best fixative is a saturated solution of sublimate in sea-water to 
every 100 ccm. of which 3 to 5 drops of glacial acetic acid have been 
added ; Flemming’s and Hermann’s fluids also gave good results. After 
fixation the films are washed in water, next in alcohols of increasing 
strength (30°, 50°, 70°, 96°, 100°), and then the preparations are ready 
for staining. The time required for immersion in the fixative is from 
* Ann. Inst. Pasteur, xii. (1898) pp. 801-3 (3 pis.). 
