is dried on a cover glass, and "fixed" by heating the cover, but not too strongly. The cover is next transferred to "chloroform-methylene-blue" (12 to 15 drops of a saturated alcoholic solution of methylene-blue, to which has been added 3 to 4 c.c. of chloroform); after 4 to 6 minutes, according to the thickness of the film, the cover is removed, the chloroform allowed to evaporate spontaneously, and the excess of coloring matter removed by rinsing in water. With fresh milk only the bacteria are stained dark-blue; if the milk has curdled, however, or become sour, the casein particles will be found stained pale-blue, which will not interfere with the visibility of the dark-stained bacteria.-Microscopical Bulletin, 1892, 21; from Centralb. Bacterienkunde, 1891.

Bacterium Amylozymicus.-L. Perdrix has separated from Paris water a bacillus, B. amylozymicus, which ferments starch, with production of amyl alcohol. It is separated by cultivation on potatoes, and finally on gelatin. The bacillus is 2-3 μ long, and 0.5 μ thick; the rods are joined in pairs and chains, and in the absence of oxygen are motile, like Vibrio butyricus, Pasteur. The rods are readily stained; the spores are set free through the dissolution of the walls of the mother cell. The bacillus flourishes only in the absence of oxygen, readily, however, either in a vacuum or in hydrogen, nitrogen, or carbonic anhydride. The optimum temperature is 35°; it grows quite well at 20-25°; at 16-17°, fermentation commences at the end of four days. Its "maximum" temperature is 42-43°. It will grow in all the usual cultivating media, ferments the sugars and starch, but does not attack cellulose or calcium lactate, differing in this respect from Vibrio butyricus, Pasteur. Acids are produced during the fermentations which it causes, and the presence of acidity, equivalent to 0.055 gram sulphuric anhydride, or of alkali equivalent to 0.08-0.11 gram in 100 c.c., is sufficient to arrest the process; the addition of calcium carbonate to the liquid enables the fermentation to become perfect. Glucose ferments to hydrogen, carbonic anhydride, acetic and butyric acids during the first three days; from the third to the ninth day no acid is formed. From saccharose and lactose acetic acid is formed during the first five days. From the fermentation of starch a distillate was obtained, of which onethird was amyl alcohol, and from 100 grams of potatoes, 2.3-2.5 c.c. of alcohols were separated. The sugar obtained from starch is very similar to glucose, but has a less rotatory action, and its phenylglucosazone melts 10° lower than that from glucose; 94 per cent. of the starch is converted into sugar, carbonic anhydride, ethyl and amyl alcohols, acetic and butyric acids, and 6 per cent. is converted into dextrin. The sugar formed by the bacillus from starch may be fermented perfectly with beer-yeast, either after sterilization, or in the presence of the bacillus. If either the sugar obtained by fermentation of starch with this bacillus, or a sterilized mash, be fermented with a pure cultivation of yeast, no fusel oil is formed, and the author concludes that the fusel oil found in commercially prepared

alcohol, is formed by the action of bacteria. The B. amylozymicus remains uninjured for 10 days at 50-55°.-Am. Journ. Pharm., 1892, 152, from Journ. Chem. Soc., 1892, 90.

Microbes in the Air.-E. W. Lucas gives a sketch of the apparatus, which consists of three flasks (say half gallon, flat bottomed). One, A, to act as the air receiver, the second, B, is filled with water and connected with the

first, while the third, C, is connected by a syphon with the second, to receive its contents. Flask A is prepared by careful sterilization, then 20 c.cm. nutrient gelatin is introduced and the mouth plugged with sterile cotton, after which it is heated in a steam sterilizer for a half hour on three successive days. A two-hole rubber cork which has been previously sterilized and fitted with one tube reaching nearly to the bottom and its top end drawn to a fine point and sealed, the other fitted with a tube bent at right angles at each end to connect flasks A and B. The flask B is filled with water and connected by a syphon with flask C, which stands on a lower level. When ready for work the upper end of glass tube in flask A is broken off to leave a small orifice, and the syphon arranged so that water will fall by drops into C, so that fresh air enters A slowly. When the water flask has emptied itself, flask A is disconnected and rubber caps placed over the tubes and the flask set aside to allow the colonies of microbes to develop. Pharm. Journ. Trans., July 1891, 63.

BILE AND URINE WITH CONSTITUENTS.

Chemistry of the Liver.-A. P. Luff has read a paper on this subject before the Chemists' Assistants' Association, which is an excellent resumé of the whole subject, but contains nothing especially new, and cannot well be condensed. Reference must be had to Pharm. Jour. and Trans., April 1892, 884, or Am. Jour. Pharm., 1892, 322-328.

Bile-Influence of Purgatives.-Loewenstein found that large doses of aloes, rhubarb, cathartic acid, jalap, gamboge and podophyllotoxin do not increase the biliary secretion; on the contrary, the last two drugs lessen it. Absence of bile in the intestines lessens the purgative effects of gamboge, jalap and podophyllotoxin, and increases the action of rhubarb and aloes. -Am. Jour. Pharm., 1892, 288, from Bull. Therap., 1891.

Influence on the Action of Pancreatin.-See under Pancreatin.
Bile-Detection in Urine.-See under Urine.

URINE.

Urine-Testing.-Louis Siebold calls attention to several points often

overlooked.

Albumen.-In incipient cases of albuminuria and Bright's disease, the urine collected the first thing in the morning is occasionally quite free from albumen, though portions collected later on may contain appreciable quantities. After the administration of acids the urine fails to respond to some of the tests. The nitric acid test is the least delicate test; in every case where this test appears to indicate albumen, while the heat test and the potassium ferrocyanide (with acetic acid) test give negative results, the nitric acid indication should be regarded with distrust. After the administration of balsams the urine reacts with nitric acid as an albuminous urine; the resinous precipitate, however, is soluble in alcohol and in excess of nitric acid. The heat-test should be performed with a filtered urine, and a test-tube with unboiled urine should always be used for comparison. The same precaution should be observed in applying the ferrocyanide test. The several tests Siebold arranges in the following order: The most delicate being picric acid, then potassio-mercuric iodide, ferrocyanide, heat, cold nitric acid, and last Jolles' bleaching-powder test.

Bile.-Siebold considers Rosenbach's test the most reliable: Filter the urine through white filtering paper, and let a drop of concentrated nitric acid run down the side of the still moist filter. It leaves a yellow streak which soon turns orange, with a violet border, outside of which blue and emerald-green zones will be observed.

Blood. He recommends as a chemical test that of Luchini: Shake the urine with chloroform and one drop of acetic acid, when the presence of blood may be deduced from the color imparted to the chloroform.

Sugar. The best way in which to test for sugar is the following: Heat

two fluid drachms of Fehling's solution in a test tube to boiling, and add 5 to 10 drops of urine; if the sugar be abundant the well-known deposit will be observed. Traces of sugar are detected by adding 11⁄2 fluid drachms of the urine to the hot solution, heating again to boiling, and allow it to stand for some time. If no milkiness is produced as the mixture cools, the urine is either free from sugar, or it contains less than per cent. If the quantity of sugar is very small, the mixture loses its transparency, and passes from clear greenish-blue to a light greenish opacity, as if a few drops of milk had been added.

Quantitative Examination.-Quantitative analysis of samples of urine not representative of the whole day's discharge are in the majority of cases of but little practical value.-Chem. Drug., Jan. 1892, 105.

Urine Test for Albumen in Presence of Biliary Matter (Icteric Urine). -Grocco finds that the usual reagents for albumen in icteric urine at times produce a precipitate similar to coagulated albumen, but being soluble in alcohol, and not giving the biuret reaction. To avoid being misled, it is necessary to treat the urine with or so of its volume of concentrated acetic acid, and put it aside for six to eight hours at a low temperature; it is then filtered, and the usual albumen tests applied. The author finds this pseudo-albuminous precipitate to be composed of biliary pigments, principally biliverdin.-Am. Journ. Pharm., 1892, 316, from Rep. Pharm., 1892, 168.

Urine-Estimation of Albumen.-Jolles considers the acetic acid potassium ferrocyanide test as the most delicate, especially if the urine be filtered before applying the test, and the mixture compared with the clear urine; in this manner it will be possible to detect albumen even if present in traces amounting to only 0.0008 gm. per 100 c.c. Bacterial urine cannot be rendered clear by filtration, but must be shaken up with a little infusorial earth, when a clear filtrate can be obtained. Traces of albumen, adhering to the earth, can be removed by washing the earth with a warm solution of potassa.-Year-book Pharm., 1891, 119, from Zeits. analyt. Chem., xxix., 407.

Urine-Test for Albumen.-E. Spiegler states that the following reagent is more sensitive than potassium ferrocyanide with acetic acid. The reagent consists of a solution of 8.0 mercuric chloride, 4.0 tartaric acid, 20.0 sugar in 200.0 water (all in gm.)

A test-tube is filled to about one-third or one-half with the reagent, and the previously filtered urine, to which has been added a little concentrated acetic acid, is added slowly, drop by drop, so as to form a layer on top of the reagent. Spiegler states that less than 1 : 50,000 of albumen can be detected in this way by the formation of a white ring in the line of contact. The addition of acetic acid is made to prevent the formation of mercuric carbonate.-Zeits. Oesterr. Apoth.-Ver., 1892, 65, from Wien. klin. Woch.S., 1891.

Urine-Optical Estimation of Albumin.-H. O. C. Ellinger has recently found that albumin may be estimated by Jean's oleorefractometer.

The apparatus consists of a tube, having a telescope at one end and a lens at the other, which is placed in front of the source of light. In the centre of the instrument is a reservoir confined between two glass plates, and within this reservoir is a removable prism with glass sides. If both the reservoir and the prism are filled with the same liquid, the observer looking through the telescope will find, upon a photographically reduced scale contained inside of the tube, a shadow, the perpendicular line of which coincides with the mark o. If the two liquids differ optically, the shadow will begin at a different mark. In some cases it will be to the left, in others to the right of zero. According as the case may be, there is an arrangement by which the observation may be easily made in either case.

Ellinger has found that if urine containing albumin is deprived of the latter by heating, addition of a drop of dilute acetic acid, and filtration, a portion of the filtrate then introduced into the central reservoir of the instrument, and some of the unboiled but filtered urine introduced into the prisms, the shadow will be pushed to the right in proportion to the amount of albumin. In five different experiments the author obtained the following readings on the scale:

By actual analysis the corresponding amounts of albumin in 1,000 parts of the urine were found to be:

2.71,

4.36,

4.94,

5.10,

5.22.

It will be seen, therefore, that the results are sufficiently accurate for clinical purposes.-Am. Drug., 1891, 345; from Jour. f. prakt. Chem., 1891, xliv., 256.

Urine Test for Albumen.-J. A. MacWilliam recommends salicylsulphonic acid as a reliable test. To 20 minims of urine in a very small test tube add a drop or two of a saturated aqueous solution of the acid (if the urine is strongly alkaline, an extra drop or two of the solution should be added). Shake the tube quickly, and examine at once. The appearance of an opalescence or cloudiness immediately or within a very few seconds is a test for proteids (if the opalescence occurs first in a couple of minutes, it indicates traces only of proteid). If on heating to boiling the opalescence or precipitate does not disappear, it is due to albumen; if on the other hand it clears up on heating to reappear on cooling, it is due to albumoses or peptones.-Am. Journ. Med. Sci., July 1891, 192, from Brit. Med. J.; Am. Journ. Pharm. 1891, 446.

J. Neumann recommends strongly salicylsulphonic acid as test for albumen, because it can be used in the cold, and is as sensitive a test as any other.-Pharm. Centralh., 1892, 31.