— Book. Copyright N° COPYRIGHT DEPOSIT: THE Examination of the Urine OF THE HORSE AND MAN BY PIERRE A. FISH, D.Se., D.V.M. PROFESSOR OF VETERINARY PHYSIOLOGY NEW YORK STATE VETERINARY COLLEGE CORNELL UNIVERSITY THIRD EDITION REVISED Comstock PusuisHinc Co., AGENTS Irnaca, N. Y. 1919 Copyright 1919 by PIERRE A. FISH” PREFACE. This manual represents the published form of mimeographed sheets, which the writer has used, for some years past, in giving instruction to his veterinary students. Data relating to the urine of the horse is not abundant and, unfortunately, some of the tests which may be standard for human urine are not reliable for that of the horse and, therefore, require modification. In order to emphasize this fact, the writer has employed the comparative method and asks the students to examine their own urine along with that of the horse and note carefully the differences in the results. The importance of urine examination for diagnosis and prognosis in human medicine is too well known to require emphasis here; but, notwith- standing certain difficulties in the way of its application in veterinary medicine, the writer believes that there is no important reason why just as much valuable information may not be derived, in certain cases, from the urine of the horse by an up-to-date veterinarian, as the physician obtains from the urine of his patients. Simplification of methods, without too great a sacrifice in accuracy, is essential for satisfactory examination,—especially by veterinarians. The writer realizes that only a short step has been taken in this direction, but hopes that time will lighten the difficulties. Beas OcToBER, 1906. PREFACE TO THE SECOND EDITION In the present edition some changes have been made in the text and new material added with the hope of bringing the manual! more completely up to date. There is still much to be done in investigating and simplifying the tests applicable to the urine of the domesticated animals and it is to be hoped that more research may be developed along this line. BAG Ey. NOVEMBER, 1911. PREFACE FOR THE THIRD EDITION Experience has shown the desirability of making certain changes and some additions to the previous edition in order to bring this work up to date. Acknowledgement is gladly made to Dr. C. E. Hayden and former students whose suggestions and interest have been of much value Marcu, 1919. Poa, (3) TABLE OF CONTENTS. CHAPTER I. The Secretion ofiimine sc. sc fs. teases nati ts techs, oom Wee pp. CHAPTER II. Quantity, Color, Transparency, Consistency, Reaction, Degreesor Acidity, SpectielGravity..... > .0. ose pp. CHAPTER III. Qualitative tests: Inorganic Constituents, Water, Chlorides, Sulphates, Phosphates, Carbonates....pp. CHAPTER IV. Organjc Constituents: Urea, Uric Acid, Hippuric Acid, Creatinin, Mucus, Indican, Oxalic Acid, Acetone, Diacetic Acid, Urobilin, Leucin, Tyrosin, EARGTION sc Sheree trees 1, ok I coche Re ee pp. CHAPTER \V. Abnormal Substances in the Urine: Albumin, Sugar...pp. CHAPTER VI. Bile MB loods Misia 3... Seep Ri ones Aree eee pp. CHAPTER VII. Quantitative Analysis, Centrifugal Method: Phos- phates, Chlorides, Sulphates, Uric Acid, Uricom- eter, Urea, Albumin, Saccharometer, Sugar, Ehr- heh's MDiaZO-TeaACtiOn otk. See acne re Poss ng cee pp. CHAPTER VIII. Volumetric Methods: Chlorides, Phosphoric Acid, Sulphu- ric Acid, Relation of Constituents in Normal Urine pp. CHAPTER IX. Chemical Examination of Urinary Deposits, Acid Urate of Soda, Uric Acid, Cystine, Pus, Blood, Al- kaline Urates, Triple Phosphates, Calcium Oxalate, Calcium Casbonate, us Blood 2. scene ese ee pp. CHAPTER X. Microscopical Examination of Urine, Unorganized Sedi- ments: Uric Acid, Acid Urate of Soda, Oxalate of Lime, Hippuric Acid, Calcium Sulphate, Calcium Phosphate, Triple Phosphate, Urate of Ammonium, Cystin, Leucin, Tyrosin. Organized Sediments: Epithelial cells; Glycogen cells, Amyloid bodies, Hyaline, Granular, Epithelial, Fat, Hemorrhagic and Waxy casts; Cylindroids; Blood and Pus cor- puscles; Spermatozoa’ “Mucus; “Bactertag ac. 2.23% pp. Form for Urine Examinations, Procedure in Examining a sdiiple Obvurine .. Set ott Ae een cd pp. Appendix, Formulae for Reagents. .......0...0-0..5... pp. 7-12 12-19 19-24 24-33 33-41 41—45 45-56 56-59 URINE ANALYSIS. APPARATUS FOR THE LOCKER 1 dozen test tubes, 6 inch 1 dozen test tubes, 5 inch 1 Minim pipette 1 Beaker, 10 oz. 1 Graduate, 30 cc. 1 Graduate, 250 cc. 1 Flask 1 Funnel, 1% inch 1 Funnel, 3 inch 1 Watch glass 1 Evaporating dish, 8 oz. 1 Urin>meter 1 Glass rod 1 Thermometer 1 Crucible, 8 cc. Special apparatus, not found in the locker, may be obtained, when needed, 1 Piece wire gauze 1 Piece absorbent cotton 1 Box matches 1 Test tube brush 1 Test tube rack 1 Test tube holder, wire 1 Tripod 1 Piece muslin 1 Pack filter papers, 3 inch 1 Pack filter papers, 6 inch 1 Sponge 1 Clay triangle 2 Tin cans 1 Copper water bath 1 Towel by handing an order for it to one of the assistants. (5) PLATE 1. SCHEMATIC SECTION OF THE KIDNEY 1. Cut portion of the Kidney. 2. Pyramid. 3. Papilla of the Pyramid. 4. Glomerulus, enclosed in Bowman’s Capsule. 5. Bowman's Capsule. 6. Convoluted portion of Renal Tubule. 7. Loop of Henle. 8. Collecting Tubule. 9. Capillary net work,—second set; the first set is the Glomerulus. 10. Openings of Collecting Tubules on Papillae, from which the Urine drops into the Pelvis of the Kidney. THE SECRETION OF URINE. I; The blood supply to the kidneys is an important consider- ation in the secretion of urine. At the outset, one is impressed by the fact that a relatively large artery supplies a relatively — small gland. The blood pressure of the renal artery is nearly, if not quite, as great as that of the abdominal aorta, which means that about as much pressure 1s required to force the blood through the kidneys as is required to send the blood through the pedal extremities. The renal vein is also a relatively large vessel as compared with the size of the kidney or with the efferent vessels of other glands. The pressure within the renal vein is very low, practically as low as that in the posterior vena cava with which it connects. The artery, after entering the kidney, breaks up into branches which pass between the pyramids. At the junction of the cortex and medulla these branches form arches in the substance of the kidneys and from these arches branches run outward into the cortex to supply the glomeruli; other branches pass inward to supply the pyramids. Each glomerulus has its afferent and efferent vessel and of these the efferent is smaller. On issuing from the capillaries of the glomerulus, the efferent vessel soon breaks up into a second set of capillaries which sup- plies the uriniferous tubules; so that in this arrangement the blood goes first to the glomeruli and later supplies the tubules. The blood from the second set of capillaries is finally gathered up by'small vessels which unite to form ultimately the renal vein. The glomeruli and the uriniferous tubules are the portions of the kidney actively concerned in the production of its secretion— the urine. The following points are therefore worthy of special notice; in the malpighian body the arterial blood in the glomerular capillaries is separated, from the inside of the capsule of Bow- man by a thin layer of flattened epithelial cells only; two sets of capillaries exist, one set forming the glomeruli and the other supplying the uriniferous tubules, the blood supplying the tubules (7) 8 must have first passed through the glomeruli, and is therefore more concentrated; in certain portions of the tubules short cylindrical epithelial cells are found which are comparable to true secreting cells found in other glands; and finally, the smaller size of the efferent as compared with the afferent vessel fulfills a condition which retards the flow of blood through the glomerulus. The conditions are favorable for a high and variable pressure in the glomerulus and for a lower and more constant pressure in the second set of capillaries around the uriniferous tubules. On account of the resistance offered by a double system of capil- laries the blood pressure in the kidneys is kept relatively high. The changes in blood pressure may be observed upon the Cracce oF URINE Fie. 1. Artery. 2. Glomerulus. 3. Capsule of Bowman. 4. Convoluted portion of Tubule. 5. Capillary net work. 7. Loop of Henle. 8. Collect- ing Tubule. 9. Opening of Tubule on Papilla. kidney itself by means of an apparatus known as the oncometer. With each rise in the blood pressure the kidney swells, with each fall in pressure it contracts, and a tracing can be obtained very similar to an ordinary blood pressure tracing. The glomeru- lus suspended in its capsule is also influenced by these changes. When the capillaries are engorged with blood the glomerulus fills the capsule, when collapsed there is an evident space between the membranes of the glomerulus and the capsule. In 1842 Bowman advanced a theory relating to the pro- duction of urine which, while recognizing to some extent certain physical factors, also brought out the view that the urine is a secretion. In 1844 Ludwig advanced the theory, sometimes referred to as the mechanical theory, in which only physical processes were involved, 7. e., filtration, diffusion and osmosis. It is true that in the capilfaries of the glomerulus there is a high resist- ance because of the smaller size of the efferent vessel, there is, therefore, a higher degree of pressure in the glomerulus than in the capsule in which it is suspended. This inequality of pressure is favorable to filtration. As the result of any filtra- tion from the glomerulus some of the water of the blood with a certain proportion of dissolved substances would pass into the beginning of the uriniferous tubule. The effect of such a filtra- tion would render the blood remaining in the glomerulus and second set of capillaries more concentrated, and in the second set of capillaries in connection with the uriniferous tubules the essential elements of an osmometer would be obtained,—an animal membrane formed by the delicate wall of the capillary and the wall of the tubule, upon one side of which there is a dense fluid, the blood, and upon the other a weak saline solution, condi- tions which are favorable to the processes of osmosis and diffusion. In this theory, as a result of the interchange of these fluids, some of the water passes from the tubule to the blood, making it less concentrated, and the products of retrogressive meta- morphosis,—urea and other organic substances,—pass from the blood to the tubule, forming urine. An objection in connection with the diffusion of urea has been raised in that it is a well-known fact that the urine contains a much greater amount of urea than does the blood and that it would be contrary to the laws 10 of osmosis and diffusion for a fluid weak in urea (the blood) to pass through a membrane a substance which has accumulated in greater amount in a fluid (the urine) on the other side of that membrane. It should also be remembered that the blood is constantly moving and pre- sumably the fluid in the tu- bules:is doing the same. In other words, it would seem that if the conditions indicate an osmometer, it varies from experimental ones in that the fluids on either side of the membrane are moving in a definite direction. The pro- teid constituents do not nor- Fic. 2 mally leave the blood on ac- 1. Artery. 2. Afferent Vessel. 3. count of their well-known in- Glomerulus. 4. Capsule of Bow- a ta re 3 man; 5. Biferent-Vessel. 6. Capi: SSPOSiViOA La OSMosIs. lary network. 7. Uriniferous Tub- One important fact, however, a ee ee remains unaccounted for in Ludwig’s mechanical theory, and that is, if the uriniferous tubules are stripped of their epithelial cells, as they often are in disease, urea and some other nitrogenous products are no longer, or only imperfectly eliminated, and become stored up in the blood and produce the condition known as uremia, al- though the conditions of an osmometer remain. It must, there- fore, be admitted that there is some direct or elaborating action on the part of the epithelium as originally suggested by Bowman, although under normal conditions, transudation, diffusion and osmotic processes may occur coincidently. The theoretical conclusions of Bowman have been con- firmed and extended by the practical researches of Heidenhain, who injected indigo-carmine into the blood of animals and found that it was promptly removed by the kidneys. These organs were removed at suitable intervals after the injection and care- fully examined under the microscope. In no instance did he find any of the indigo in the capsules of Bowman, but it was found abundantly in the cells lining certain portions of the tubules, and in the lumen of the tubules near these cells. Similar experiments 11 with the urate of sodium showed that it likewise was secreted at the same place and in the same manner. Further investigations have tended to diminish the impor- tance of the “‘mechanical”’ factor and to develop a selective or secretory function for the cells, even including those of the glomeruli, by virtue of which the cells will permit certain sub- stances to pass through and prevent others, among the latter, albumin. The view is advanced that the passage of the fluid through the glomerulus is not a mere transudation, but a matter of selection also. The selective action of the renal cells is both qualitative and quantitative. As an example of the former, it is shown by experiment that if some egg albumin be injected into the blood, it is promptly eliminated by the kidneys. Egg albumin is not very markedly different from the serum albumin of the blood; both are indiffusible, but the renal cells recognize the former as a foreign substance and immediately separate it from the blood. The sugar, in normal quantity in the blood, although a diffusible substance, is not selected. Urea, which is also dif- fusible, but existing in the blood in much smaller amount than sugar, is selected and appears in the urine. Why this should be so it is difficult to explain, although it is a well-known fact that the sugar serves as a food for the tissues and is needed by the system, while urea is a waste product and would be detrimental to the system if not eliminated. That the cells exert a quantitative selection is shown by the fact that when sugar is present in the blood in excessive amounts, 3 parts per 1000 or over, the excess is promptly elimi- nated. Diet influences the reaction of the urine. A vegetable diet favors an alkaline reaction; a diet of flesh favors an acid reaction. The urine of the herbivora is therefore alkaline, while in the carnivora and omnivora the urine is mainly acid, although influenced to some extent by the kind of food eaten. The forma- tion of an acid fluid from alkaline material,—blood and lymph,— is at first sight puzzling. The well-known fact that the gastric secretion of all mammals is acid is a case in point. Further- more, experimental evidence shows that if an alkaline solution of sodium bicarbonate be placed upon one side of the membrane in an osmometer andasolution of neutral sodium phosphate be 12 placed upon the other side and a weak electric current be sent through the solutions, the fluid in contact with the positive pole will, in a short time, become acid from the formation of acid sodium phosphate, while the fluid in contact with the negative pole is increased in alkalinity Na HCO;+ Naz HPOs;= Naz CO;+Na He PQ,. While it may be true to a considerable extent that the kid- ney merely removes certain constituents from the blood and transfers them to the urine, it has been shown that the activity of the renal cells is required in the production of hippuric acid, —a new product, as hippuric acid is not present in the blood. In general, the theory explaining the secretion of urine, according to observed facts, is ’one which, while recognizing the process as partly physical, also requires some process of activity or elaboration on the part of the kidney itself. EH. The urine of all mammals may be regarded, for the most part, as a solution of constituents derived from the metabolism of the tissues of the body. Some of these constituents, espec-— ially the inorganic, may appear in the urine in the same form as they are taken into the body in the food, e. g., sodium chloride; others, especially the organic, represent decomposition products derived from the food or tissues, e. g., urea, creatinin, etc. The composition of the urine may, therefore, to some extent be regarded as an index of tissue activity, and the examination of this secretion is of considerable importance in clinical diagnosis and prognosis. That there is a relationship between the diet and the renal excretion is shown by the examination of the urine of the three great classes of animals grouped according to the food they eat; herbivora, omnivora and carnivora. Perhaps the first and most striking characteristic of the urine of each group is its chemical reaction. In vegetable feeders it is normally alkaline; in flesh eaters it is markedly acid, and in the omnivora it may be acid or alkaline according to the preponderance of the fleshy or vegetable material in the food. : The relative proportion of phosphoric and carbonic acids 13 in the blood depends very much upon the composition of the diet. The chief mineral constituents of animal. food are phosphates; those of vegetable foods carbonates. The blood of herbivorous animals is therefore rich in the latter, that of the carnivora in the former, and the acids formed in the tissues will form bicarbonates in the blood of the herbivora, and acid phosphates in that of earnivora. Acid phosphates hold earthy phosphates in solution, and bicarbonates dissolve. earthy carbonates. Hence the urine of carnivorous animals is rich in phosphates of lime and magnesia, ~ that of the herbivora in calcium and magnesium carbonate. In the practical treatment of this subject it is convenient to regard the horse as the type for the herbivora; man for the omnivora; and the dog or cat for the carnivora. [In all cases throughout this work it is to be understood that, unless especially directed, the same tests are to be performed upon the urine of man and horse and a parallel record of such tests, for comparison, is to be kept upon the blank pages. : In man, fresh, normal urine is a clear, golden-colored, trans- parent liquid, having a peculiar aromatic characteristic odor, and a bitter saline taste. : In the horse, the urine is of a yellowish color when passed, but turns to a deep brown color upon standing for a time, due to the oxidation of pyrocatechin. It is more or less turbid and of a viscous character. Its odor is somewhat ammoniacal and strongly aromatic and more penetrating than that of man. The urine is chiefly a solution of urea and certain organic and inorganic salts; epithelial cells and mucus may also be held in suspension. Like milk and other animal fluids, the urine is not of constant composition. It is influenced by various factors, such as food, the amount of water or- other fluids taken into the body; the temperature of the skin; the emotions; blood pressure, general or local; exercise; the time of day; age; sex; and medicines. Quantity. The amount varies considerably. In man, the quantity for twenty-four hours ranges from 1000 cc. to 2000 cc. In the horse, the average amount is about 3000 cc. to 4000 cc., although it may go as high as 7000 c.c. or 9000 cc. In the ox a still greater quantity is secreted, the usual limits being from 4500 cc. to 19000 cc. In the sheep, from 250 cc. to 700 cc. In 14 the pig it varies from 1200 cc. to 6000 cc. In the dog it varies from 200 cc. to 900 cc., depending upon the size of the animal. Color. The color ranges from pale yellow to brown. The normal color of urine is due to pigments probably derived from the coloring matter of the bile. Transparency. The urine of the horse is normally more or less opaque, that of man should be transparent at the time of passing. Many pathological urines, however, are perfectly ~ clear. Pathological turbidity may be due to urates, phosphates, pus, bacteria, spermatozoa, fatty globules, blood, etc. All urines become turbid after standing for a time. The urine of the horse is especially turbid at the end of micturition, exceptionally it may be clear but becomes turbid on cooling. Its opacity is due to the presence of earthy salts of the carbonate of lime precipitated and formed by the dis- engagement of a certain amount of carbon dioxide from the bicarbonate of lime. The turbidity increases when the urine remains for any length of time in the bladder; it reaches its maximum when the urine is cooled by exposure to the free air; it diminishes after the ingestion of a large quantity of water. A clear, limpid urine is generally pathological in the horse; it indicates polyuria and the reaction in this case is usually acid, exceptionally neutral or alkaline when the phosphates are modified qualitatively or quantitatively. The turbidity of horse urine is abnormal when it is due to the presence of the phosphate of lime, or of calcium sulphate, or acid salts; also from the existence of albuminoid substances, (exudates, leucocytes in interstitial nephritis). | In other animals the passing of turbid urine is usually re- garded as abnormal. Consistency. In the horse the urine is very viscid on account of the contained mucus and sometimes of epithelial debris. It filters very slowly. Urine taken directly from the ureter or the pelvis of the kidney is still more viscid, having a consistency very similar to that of egg albumin and a specific gravity some- what higher than normal. Reaction. The reaction of human urine is acid, that of the dog more so than that of man. In the pig it is sometimes acid and sometimes‘ alkaline, depending upon the diet. In the horse 15 and sheep it is alkaline, also in the ox, but in the calf and foal while suckling itis acid. Herbivorous urine is alkaline, but if such ani- mals are starved for a time they practically become carnivorous in that they are living upon their own tissues and under such con- ditions their urine becomes acid. In the herbivora the reaction is normally alkaline and is due to the presence of bicarbonates or carbonates of lime or potassium. It would appear that the salts present in the vegetable food undergo oxidation to form organic acids, and these in turn are transformed into bicarbonates or carbonates, causing alkalinity of the urine. In man the acidity is due to the presence of acid sodium phosphate NaH, PO,. The urine passed before breakfast and during fasting or perspiration is more acid than at other times; during digestion and after meals the acidity is decreased. The degree of acidity of the urine may be determined by the use of the acidimeter, a graduated glass tube devised by Dr. H.R. Harrower. His description of it and its use follows: “The acidimeter consists of a glass tube so graduated that 10 ce. is the first measuring point. From this upward the tube is graduated in fifths of degrees up to 100°, each degree repre- senting the amount of decinormal sodium hydroxide solution required to neutralize 100 cc. of urine. The method of using © the acidimeter is as follows: The tube is filled with the urine to be tested, until the lower edge of the meniscus is just on the 10 cc. mark. Two drops of phenolphthalein indicator solution are added, and then with an ordinary medicine dropper deci- normal sodium hydroxide solution is slowly added, inverting the tube after each addition, until the color of the fluid has just .been changed from a yellow to a light rose pink. The acidity in degrees is now read off on the tube at the level of the fluid. The normal acidity of a mixed 24 hour specimen should be between 30 and 40 degrees. (With very concentrated urines in which the acidity is above 100° the tube may be filled to the 5 cc. mark and water to the usual level. The resulting figures are, of course, doubled.) If the urine is alkaline in reaction and it is desired to esti- mate the degree of alkalinity decinormal hydrochloric or oxalic acid solution must be used in place of the sodium hydroxide, the pink color present being just discharged by the acid.” 16 The ‘‘Acid Index’ or ‘Acid Unit” may be obtained by multiplying the degree of acidity by the amount of urine passed in 24 hours. The normal man is about 40,000 acid units. In man there is increased acidity physiologically during the night; with a flesh diet; after strong muscular exertion; during the intervals of gastric digestion; after the ingestion of mineral acids. There is increased acidity, pathologically, in fevers; in rheu- matism; after asthmatic attacks; in emphysema, pneumonia — and pleuritis. The urine is less acid, or alkaline, physiologically, during gastric digestion; after hot or prolonged cold baths; after pro- fuse sweating; after copious ingestion of vegetable acids and their salts. : Pathologically in acute and chronic inflammation of the urinary tract as in cystitis; in decomposition of the urine in the bladder in retention; in some cerebral and nervous diseases: in anemia; chlorosis; and general debility. When the urine of man is set aside in a cool place it grad- ually becomes more acid. This is called acid fermentation. After longer exposure to a warm atmosphere the urine becomes neutral, and finally strongly alkaline in reaction. It becomes turbid, has an ammoniacal odor, and deposits triple phosphate, ammonium urate, and great numbers of microbes exist. This is alkaline fermentation and is due to the transformation of the urea into ammonium and carbon dioxide, by means of a ferment produced by an organism known as the muicrococcus ureae. (Fig. 20). This organism is said to be conveyed through the air and to exist commonly around the orifice of the urethra. As long as the urine is acid the organism does not exist in the bladder, but may sometimes gain entrance through the medium of a sound or catheter. Test the reaction of the urines with red and blue litmus paper. Some urines change both the red and blue paper and are termed amphoteric. (In taking notes of the experi- ments it is well to record the results in parallel columns, the horse urine in one column and the human urine in the other). Specific Gravity. The specific gravity of human urine ranges from 1015 to 1025, the average being 1018 to 1020. That th ; Bb A Mt ay, of the horse ranges from 1020"to 1050, the average being about 1035. That of the cow is lower, ranging from 1015 to 1045. It seems to depend considerably upon the milk secreted. In milch cows the urine contains a greater amount of water and a lesser amount of solids. The specific gravity of the urine of the sheep ranges from 1015 to 1060; that of the pig from 1005 to 1025, and of the dog from 1016 to 1060, depending upon the diet. Cat, 1020 to 1040. The specific gravity may be ob- tained in different ways. The sim- plest and most usual way is to employ the instrument known as the urin- ometer. Some urinometers are not strictly accurate, but they may be tested by filling the urinometer jar with distilled water at 15° C. (60° F.) Read the division of the scale corre- sponding with the surface of the fluid looking above or below the meniscus as is found to be the most correct for the zero reading. Always adhere to this method when using the same urinometer. Test the spe- cific gravity of the urines and make the necessary corrections. If the urine is warmer than 15° C. add 1 to the last right hand figure of the spe- cific gravity for every 4 degrees of C. temperature, or for every 7 degrees of extra F. temperature. As oppor- tunity presents, test the specific grav- ity of some warm, freshly passed urine. Test the same urine later, when cool, and note if any difference — in the reading. Urinometers already corrected for the ordinary room tem- perature (70° F.) may be obtained, in which case the temper- ature corrections may be omitted. If the urine should be too dense to read easily on the urinometer, dilute it with an equal volume of distilled water and multiply the reading by two to Rie. 3 Urinometer. i be van} Ly iw? ded pe a Tye ats it i ‘ he Le ri 18 get the correct specific gravity. The variation of the specific gravity depends upon the amount of the solids in the urine. The amount of solids may be estimated with approximate accuracy from the specific gravity by Christison’s formula (Haser-Trapp’s coefficient): ‘‘Multiply the last two figures of a specific gravity expressed in four figures by 2.33. This gives the quantity of solid matter in every 1000 parts.’”’ (The number of grams in 1000 «ce. Example. Suppose a patient passes 1400 cc. of urine in 24 hours and the specific gravity is 1020, 20*2.33=46.6 grams of solids in 1000 cc. To ascertain the amount in 1400 cc. use the following proportion: 1000 cc. : 1400 cc. : : 46.6 grams is to X (65.24 grams). The total quantity of the solids of the human urine is about 60 grams for the 24 hours or approximately AM, The following method taken from the Alkalcidal Clinic may also be used: Multiply the quantity, in ounces, of the 24-hour urine by the last two figures of the specific gravity and this by 1.1, the product will represent the total solids in grains. Thus, if the amount of urine voided in 24 hours be 36 ounces and its specific gravity 1021, the formula would be 36X21 X1.1, equal to 831 grains, the normal amount for a person weighing 100 Ibs. The amount for other weights may be determined by proportion. In general the amount of total solids is a measure (a) of the activity of tissue change; (b) of renal integrity; (c) of abnor- mal constituents in the urine. Hygienic conditions which favor “increased metabolism, as abundant food, active exercise, etc., increase the solid matter in urine, while the opposite conditions decrease them. -With the urine normal or subnormal in amounts, the solids are deficient pathologically from defective and enfeebled meta- bolism as in senility; anemia as a result of syphilis, cancer, etc.; chronic alcoholism; functional or organic diseases of the liver; from renal failure as in acute nephritis; certain’ conditions of ~ chronic renal disease; at the close of Bright’s disease; venous congestion of the kidneys, etc. With the urine increased in amount there may be a deficiency of solids in diabetes insipidus; interstitial nephritis; amyloid disease of the kidney; chronic parenchymatous nephritis. When the urine is not increased in amount, the urinary solids are increas2d in fevers; lithemia; some > 19 forms of dyspepsia. When the quantity of urine is increased the solids are increased in diabetes mellitus; phosphaturia; azoturia (excessive secretion of urea). : Calculate the solids of the urines by the formulae previously given. After obtaining the result of 1000 cc. estimate the quantity in 1250 cc. of human, and 5450 ce. of horse urine. In record- ing the tests use parallel columns, one column for the horse and the other for the human urine. III. Qualitative Tests. In all cases the urines must be filtered and perfectly clear before attempting the examination. INORGANIC CONSTITUENTS. These consist chiefly of sodium, potassium, ammonium, calcium, and magnesium, combined with hydrochloric, phos- phoric and sulphuric acids. Water. The water of the urine is derived from the food and drink, a small quantity being formed in the body. It var- ies according to the activity of the sweat glands of the skin. Chlorides. Next to the urea the chlorides form the chief portion of the urinary solids. The chlorides are increased phy- siologically after the ingestion of salt foods and much water; mental and physical activities; and during pregnancy. Pathologi- cally, they may increase after the crises of fevers; after: the absorption of exudates; in diabetes (occasionally). The chlorides are decreased pathologically, in all acute fevers; pneumonia (often entirely absent during the height of the disease); in cholera; and in most chronic diseases. An increase, or the re-establishment of the excretion of chlorides in disease is generally a favorable sign. In pneumonia it is a precursor of the crisis, and may often take place before other symptoms reveal the favorable change. Test a portion of each urine with a few drops of silver nitrate solution. A white, cheesy or curdy precipitate 20 insoluble in nitric acid indicates the presence of silver chloride. The phosphate of silver may also be thrown down but the nitric acid dissolves it, keeping it in solution. Evaporate carefully a few drops of urine upon a glass slide, with a gentle heat over the flame. Octahedral or rhombic crystals may form,—a compound of sodium chloride and urea. Examine with the microscope. (Fig. 5). Sulphates. The sulphates are chiefly those of sodium and potassium. Only a small amount of them enters the body with the food, so that they are chiefly formed from the metabolism of proteids in the body. The above are known as ordinary sulphates. Another class known as the ethereal sulphates also exist. The proportion exists in the ratio of 10 of the ordinary to 1 of the etherealin man. In the horse the proportion is about 2 of the ethereal to 1 of the ordinary. The ethereal sulphates are formed by the combination of sulphuric acid with organic bases such as phenol, skatol, etc., which originate from putre- factive processes in the intestine. The amount of ethereal sul- phates is of importance in determining whether or not the diges- tive processes are going on normally. In general the sulphates are increased physiologically by the ingestion of sulphur and its compounds; nitrogenous food; and conditions of increased meta- — bolism. After acidulating the urine with hydrochloric acid to prevent the precipitation of phosphates, add to a small part of each urine, a little 2% barium chloride solution; a precipitate of barium sulphate is formed, insoluble in nitric acid. To separate the ethereal sulphates, mix 30 cc. of urine with an equal bulk of ‘‘baryta’’ mixture. Stir and filter. This removes the ordinary sulphates (as barium sulphate), add 10 cc. of hydrochloric acid to the above filtrate, and keep in the water bath at 100° C. for an hour in the hood and then allow the ethereal sulphates to settle. This may require some little time. (Baryta mixture is prepared by making saturated solutions in the cold, of barium nitrate and barium hydrate, and adding two volumes of the meat to one volume of the nitrate). Phosphates. The phosphates consist of alkaline and earthy salts in the proportion of 2 to 1. The latter are insoluble in an alkaline medium and are precipitated when acid urine becomes alkaline. They are insoluble in water, but soluble in acids; in acid urine they are held in solution by free CO.. The alkaline phosphates (sodium and potassium) are very soluble in water, and they never form urinary deposits. The earthy are. phos- phates of calcium (Caz PO,). (abundant) and magnesium (MgHPO, plus 7H,O) (scanty). An alkaline medium precipitates them al- though not in the form in which they occur in the urine. The excretion of phosphates in the urine is largely dependent upon the amount of calcium ingested; the more calcium the food contains, the less phosphoric acid appears in the urine, and the more in the feces. ‘This is due on the one hand to the tendency on the part of calcium to form insoluble calcium phosphates in the intestinal tract and in this way to prevent the absorption of the food phosphates; on the other hand, to the well-established tendency of calcium salts to be excreted into the bowel and not into the bladder; one must imagine in the latter case that calcium salts circulating in the blood combine with circulating phosphoric acid and bear the latter with them into the bowel. The fact is of some therapeutic importance in the treat- ment of nephrolithiasis due to uric acid calculi, for the admin- istration of calcium salts in this affection by bearing much phos- phoric acid into the bowel, leads to the excretion of less phosphoric acid in the urine, and hence of normal and basic instead of acid phosphates; and as the latter precipitate and the former dissolve uric acid, it will be seen that by giving calcium we prevent the precipitation of crystalline uric acid and urate deposits in the urinary passages. (Croftan). Where considerable calcium is present in the food the excre- tion of phosphates in the urine is minimum, especially if the urine is alkaline because of the presence of sodium or potassium salts. In herbivorous animals where the urine is alkaline and where considerable quantities of phosphorus containing food are eaten, very little phosphate is excreted in the urine. Phosphates are increased in the urine pathologically in rickets: osteomalacin; osteoporosis; fractures; chronic rheu- matism; diseases of the nervous system; and after great mental 22 strain and worry. Phosphates are decreased in renal diseases and phthisis. Physiologically, variations occur chiefly from the character of the food and drink; in the horse there may be an increase in the urinary phosphates after a large feed of oats, bran, oil- cake, etc. Pathologically, they are increased during the active changes in such bone diseases as spavin, ring-bone, splint, etc. To a small amount of each urine add about half its volume of nitric acid and then add two volumes of a 5% solution of ammonium’ molybdate and boil. A canary yellow ppt. (crystalline) of ammonium phospho-molyb- date should appear in the omnivorous urine. In the herbivor- ous urine the presence of so much organic matter renders the test unreliable; although a precipitate may form it is not a typical or characteristic one for phosphates in the urine of the horse, unless the organic matter has been previously removed. To each of the urines add half its volume of ammonia and allow it to stand. A precipitate of earthy phosphates is formed, in the urine of man Filter, add enough nitric acid to give an acid reaction to the urines, and test the filtrates with ammonium molybdate as before. This method separates the earthy from the alkaline phosphates. To a portion of the urines add half their volumes of baryta mixture; a copious precipitate. Filter, add nitric acid and test the filtrates with ammonium molybdate. No ppt. should occur as the baryta mixture precipitates the phosphates as well as the sulphates and carbonates. Use a little of the magnesia mixture instead of the baryta mixture. Filter, add a little nitric acid, and test the filtrate © with ammonium molybdate. (The magnesia mixture is composed of magnesium sulphate 1 part, ammonium chloride 1 part, ammonia water 1 part, and distilled water 8 parts). To portions of the urines add a few drops of acetic acid and then a little 5% uranium nitrate solution,—a . yellow ppt. of uranium phosphate is formed. The lime, magnesia, iron and other inorganic urinary consti- tuents are comparatively unimportant, and have no special 23 clinical significance. The tests for them are somewhat compli- cated and are therefore omitted. Demonstration of Carbonates and CO, in Urine. Carbon dioxide exists in the urine to some extent in a free state. There are also various carbonates present, especially in the herbivorous urine. The amount is very variable and to a great extent is dependent on the kind of food that is eaten; large quantities of vegetable foods determine an increase both in the combined and in the free carbon dioxide. The carbonates may be broken up and COs, given off by the application of heat or certain acids. Heat experiment. Fill a small flask about half full of unfiltered herbivorous urine. Through the perforated stopper of the flask pass some bent glass tubing connected with a test tube containing lime water. Heat the urine in the flask, and as it boils the CO, will pass over into the test tube, and calcium carbonate will be formed from the union of the gas with the lime. Repeat the experiment with omnivorous urine. .» A simple proof of the presence of carbonates, or CO: in the urine is to fill a Doremus Ureometer with 25 cc. of the unfiltered urine and introduce 1 cc. of nitric acid. The larger part of the gas (COs) rises and collects in the upper portion of the ureometer. Compare the amounts thus obtained in the urines of horse and man. A simpler qualitative test is to add a few drops of nitric or acetic acid to a little unfiltered urine in a test tube. If effervescence occurs it is due to CO, set free from the car- bonates by the acid. IV. ORGANIC CONSTITUENTS. Urea is, in amount, the principal constituent of the solids of the urine. It is the most important product of the decomposi- tion of proteid in the food. In round numbers it forms from 2% to 3.5% of the urine, averaging about 2.5%. About one-half of the total solids in the urine consists of urea. Urea has no effect on litmus, it is odorless, has a weakly cool and bitter taste like saltpeter. It is very soluble in water amd alcohol, but it is almost insoluble in ether and benzine. About 90% of the total nitrogen of the urine is excreted in the form of urea. Physiologically, urea is increased by a proteid diet; exercise and muscular vigor; by drinking much water. It is decreased by fasting; non-nitrogenous food; reduction of water in the diet; alcoholic beverages, tea or coffee; indolence of mind and body. Pathologically, it is increased in all acute fevers; dyspnoea; diabetes; and phosphorus poisoning. It is decreased in uremia; acute yellow atrophy of the liver and in chronic diseases. In general, any disease that interferes with the activity of the liver decreases the urea. Any disease affecting the uriniferous tubules ~ may modify the appropriation of the urea from the blood and affect its passage into the urine. An increase of the urea inde- pendently of the physiologic variations and amount of nitro- genous food eaten is an approximate index of the amount of tissue waste in the system; on the other hand, when the urea is decreased it is an evidence of a diseased condition of the liver (the producer of urea) or the kidney (the eliminator of urea). A simple method of detecting urea is to concentrate a small amount of dog* or human urine in an evaporating dish to about half of its original volume. Place a drop or two of this concentrated urine upon a glass slide and after adding a drop of nitric acid, gently warm over the flame. If urea be present, upon evaporation, the micro- scope will show the characteristic crystals of nitrate of urea, of rhombic or hexagonal form. (Fig. 4). . *For this experiment, dog urine is more satisfactory than human because of the larger percentage of urea present. 25 Take 20 cc. of fresh, filtered dog or human urine and add 20 cc. of baryta mixture to precipitate the phosphates and sulphates. Filter, evaporate the filtrate to dryness, and extract the residue with a little boiling alcohol over the water bath very carefully. Filter off the alcoholic solution, stand the,filtrate away in a cold place for crystalliza- tion. The crystals of urea, usually long, fine, transparent needles, will separate out. Examine them under the micro- scope. (Fig. 5). Repeat in the hood, the experiment upon the urine of the horse and compare results. Heat some urea crystals in a test tube. Biuret is formed and ammonia comes off. Add a trace of copper sulphate solution and a few drops of 20% caustic potash. A rose-red color is produced,—the biuret reaction. Fic. 4 Fic. 5 Crystals of Nitrate of Urea. Crystals of Urea. Medicines which increase the amount of urea are: urea itself; uric acid, common salt, phosphoric acid, squill, theo- bromine, colchicum, cubebs, atropine, cantharides, vegetable acids, iron preparations, hyposulphite of soda, potassium chlor- ide, ammonium chloride, coca, potassium permanganate, oxygen, salicylic acid. Medicines which decrease the amount of urea are: digitalis, alcohol, coffee, tea, potassium and sodium iodides, potassium bromide, arsenic, turpentine, aklaline carbonates, mercury, anti- pyrin, valerian, quinine sulphate, benzoic acid. 26 Uric Acid, sometimes called lithic acid, is, next to urea, the most important nitrogenous constituent of the urine of man. It has been said to be absent in herbivorous urine, being replaced by hippuric. This has been shown by later researches not to be strictly true, as a trace of uric acid is found in addition to the hippuric. In birds and reptiles uric acid is the chief nitrogenous constituent, being present in greater amount than urea. In man the proportion of urea to uric acid is about 45 to 1, about 0.5 gram of the latter being excretedin the 24 hours. It causes the brick red deposit sometime seen in urine after standing fora time. Its solubility is very low, only 1 part being soluble in 14000 cc. of cold water, and 1 to 18000 in boiling. Physiologically, it is increased and diminished proportionately with urea. Patho- logically, it is increased in indigestion; acute dropsies, rheumatic and catarrhal inflammations; after attacks of gout; cancer of the liver; in leucemia; in all disturbances of the circulation and respiration. Pathologically uric acid is decreased in chronic diseases generally; diabetes and polyuria; before paroxysms of gout; anemias; chronic rheumatism; chronic diseases of spinal cord. Uric acid is generally in solution in the form of urates of sodium, potassium, ammonium, lime and magnesium. These salts are very easily decomposed even by weak organic acids. Perform the following tests upon both urines (filtered). In a conical glass, add 5 parts of hydrochloric acid to 30 parts of urine. Label and put in a cool place for 24 hours. Red or brownish colored crystals of uric acid are deposited upon the sides of the glass, or form a pellicle on the surface of the fluid like fine grains of cayenne pepper. The brownish-red color is due to pigment (uroerythrin). Murexide Test. To about 1 cc. of human urine add a little nitric acid; evaporate in a porcelain dish very care- fully to avoid charring. Cool and add a drop of dilute ammonia, a purple red color of murexide or purpurate of ammonia is formed. It turns bluer upon the addition of caustic potash solution. Dissolve a few crystals of uric acid in 10% caustic soda or potash. Add a drop or two of Fehling’s Solution— or dilute cupric sulphate and caustic potash and heat— 27 there should occur a ppt. which at first may be white and after a time turning green or reddish. (Fehling’s Solution is put up in two bottles; one labeled A, the other B. In making tests, take equal parts of A and B and add the substance to be tested. See formula in appen- dix.) Schiff’s Test. Dissolve a little uric acid in a small quantity of 10% sodium carbonate solution. With a glass rod, place a drop of silver nitrate solution on filter paper and then a drop of the uric acid solution so that the two drops partially overlap. A dark brown or black spot of reducing silver appears. Hippuric Acid, (Cy»H,NOs), is found especially in the urine of the herbivora, as the horse, ox, etc. In the urine of carnivora, and especially in that of man, it exists in but very minute quantity, usually about 0.5 to 1 gram being excreted in 24 hours. It dissolves readily in hot alcohol but is sparingly soluble in water. It occurs in man after the ingestion of certain vegetables, such as asparagus, plums, pears, and apples with their skins; a purely vegetable diet and from the use of benzoic acid, cinnamic acid, essence of bitter almonds, quinine, and analogous bodies. Hippuric acid normally seems to be derived chiefly from the husks or cuticu- lar structures of the food. Pathologically hippuric acid is increased in diabetes, chorea; jaundice and other liver complaints; and in the acid urine of x f Fic. 6 ea Gre Crystals of Hippuric Acid. Various forms of Hippuric Acid with triple phosphates. 28 patients suffering from all kinds of fevers. In testing for hip- puric acid the fresh urine should be used; if stale, benzoic acid is likely to be obtained instead. Test. Saturate the fresh urine with lime water, which transforms the hippuric acid into a salt of lime, the fluid is boiled, then filtered, evaporated to a syrupy consistency, cooled and excess of hydrochloric acid added, when hippuric acid crystallizes out on standing for 24 hours. The horse urine is to be evaporated in the hood. Creatinin (C,H;N3;0). This substance was discovered in ‘the urine by Liebig. It is easily produced from creatin, a sub- stance normally existing in plain and striated muscular tissue. Creatinin occurs constantly in normal human urine, the amount varying according to Voit, from 0.5 to 4.9 grams per day accord- ing to the quantity of proteids eaten. It is said not to be dimin- ished by fasting. Pathologically it is increased in typhoid fever; intermittent fever; pneumonia; tetanus. It is de- creased during convalescence from the above diseases; and likewise in anemia; chlorosis; muscular atrophy; tuberculosis; paralysis, Etc. Test.<, Take ’.5 .ec.. of .winem a test tube and add a few drops of freshly prepared 1% sodium nitro- prusside. Render the solution alka- line with potassium hydroxide. A ruby red color develops which soon turns yellow. The above is Weyl’s test. Acetone, if pre- sent, also gives the red reaction. Perform the above test on both urines. Caution. Creatinin’ reduces copper oxide and may be taken for small quantities of sugar. Fic. 8 Creatinin. Mucus. . Mucus in the urine is not visible, but causes cloudi- ness sometimes by entangling epithelial cells, urates, oxalate of lime and other crystals in various amounts. Add to the urine a little acetic or citric acid and, in addition a few drops of ligucr todi comp. (Lugol’s solu- 29 tion) which makes the threads or bands of mucin visible. . Indican or Indoxyl. This substance is derived from indol, one of the putrefactive products formed in the intestine. Indoxyl occurs in very small quantities in normal human urine, about .004 to .020 gram for the 24 hours. Horse’s urine is said to con- tain 23 times as much. The intestines of the herbivora are much longer than in the case of the carnivora. On this account, and in conjunction with the carbohydrate diet, a much greater fermen- tation occurs, which leads to a greater elimination of fadican in the urine. In obstruction of the intestine, or in intestinal catarrh or where the food remains a long time in the intestine and ferments there, the proportion of indican increases in the urine and causes _ a true indicanuria. Indoxyl is of considerable clinical import- ance, an increase is indicative of imperfect performance of the digestive processes. In obstructive diseases of the small intestine ' the increase of indoxyl in the urine is enormous. Pathologically indoxy1 is increased in cholera, typhoid fever, peritonitis, dysentery, Addison’s disease, cancer of the liver and stomach and pernicious anemia. Obermayer’s Reagent. This reagent has the advantage of keeping indefinitely. It is prepared by dissolving 2 grams of solid ferric chloride in 500 cc. of concentrated hydro- chloric acid. (Sp. gr. 1.19). To equal parts of urine and the above reagent add a little chloroform. Shake frequently but not too violently (otherwise an emulsion may be formed). The chloroform will become more or less blue by the indigo formed, in pro- portion to the indican originally present: According to the depth of the blue color it may be designated as little, much or copious. A test said to be somewhat more sensitive is as follows: Urine 10 cc.; 20% basic Lead acetate 2 cc. Shake and filter. Add to the filtered urine 10% Thymol in alcohol, % cc.; Obermayer’s Reagent 10 cc. and let stand for 15 minutes; add chloroform, 4 cc. Shake gently several times. Let the chloroform settle. A violet color indicates the pre- sence of indican. Jaffe’s Test. To a little urine add an equal volume of strong hydrochloric acid. Add to this mixture 2 or 3 Yreka drops of a solution of freshly prepared chlorinated soda. There soon forms a bluish cloud of indigo. Add a little chloroform and shake gently, this will take the indigo into solution and settle as a blue layer at the bottom of the test tube. The amount of indoxyl can be judged by the depth of the blue color. Indican is oxidized by free chlorine ob- tained from the chlorinated soda to indigo. Oxalic Acid. This is usually found in combination with lime in the form of calcium oxalate. The crystals are of small . size and appear in the form of dumb bells and octahedra. They occur normally in greater amount in herbivorous than in omni- vorous urine. They greatly increase after eating such vegetables as tomatoes, fresh beans, beet-root, asparagus, apples, grapes, honey, and after the use of rhubarb, senna, squills, etc. Another source of oxalic acid in the body is incomplete oxidation of carbo- hydrates and proteids or retarded metabolism. It is therefore a result of mal-assimilation and is found in dyspepsia, diabetes mel- litus, etc. The long continued excretion of an excess of oxalate of lime frequently irritates the kidneys, producing albuminuria and grave nervous disturbances and may lead to the formation of calculi: (Fig: 18.) Acetone. Normal urine may contain traces of acetone but it Occurs 1n excessive quantities as a pathological condition.* It is found in many of the fevers, certain forms of cancer, in starva- tion, and in diabetes, when it indicates an advanced form of the disease. It is associated with an increased proteid metabolism and is looked upon as a product of proteid decomposition with deficient oxidation. Lieben-Ralfe Test. Dissolve 1.3 grams (20 grains) of potassium iodide in 4 cc. (1 dram of liquor potassae). Boil in test tube, after which gently pour the urine on its surface. A yellow precipitate between the two solutions indicates an affirmative test. A more satisfactory test is to add to the urine a few crystals of iodine and of iodide of potassium with some caustic potash. Heat. Yellow precipitate—iodoform with its characteristic odor. Legal’s Test for Acetone”. Add. to 5. ce. of the urme *If pathological urine is not available, a small amount of ac2tone may be added to the urine for laboratory tests. 31 some fresh aqueous solution of sodium nitroprusside fol- lowed by a little ammonia, or sodium hydrate solution, which gives a red color. Add an excess of acetic acid and if acetone is present the red color will be intensified, if acetone is absent a yellow color will result. Compare with creatinin. These tests are not always satisfactory when applied to the ordinary urine. Greater accuracy is claimed if the urine is distilled and the tests applied to the distillate. Frommer’s Test for Acetone. To 10 cc. of urine in a test tube add some potassium hydroxide to make the urine markedly alkaline; before the latter is dissolved add 10 to 12 drops of salicylic aldehyde (made by dissolving 1 part of salicylous acid in 10 parts of absolute alcohol). Heat the mixture to about 70° C. With acetone the solution becomes yellow, then red and after long standing dark red. According to Frommer, even the minutest amount of acetone will give this reaction and no other constituent of the urine will give this color—not even diacetic acid. The reaction is explained as follows: One molecule of salicyclic aldehyde combines with one molecule of acetone to form oxybenzol-acetone. This, in the presence of strong alkalies, forms dioxy-dibenzol- acetone. The alkaline salts of this compound are intensely red. Diacetic Acid. Lindemann’s modification of Riegler’s test. Urine Lice: Acetic Acid 5 drops Lugol’s Solution 5 drops Chloroform 3 cc. With normal urine the chloroform is colored rosy red; with urine which contains diacetic acid it remains colorless. Urine which contains much uric acid should have the amount of Lugol’s Solution doubled, and should not be too vigorously shaken. Urobilin is commonly regarded as the most important color- ing matter in the urine. There is some evidence that it represents a reduced form of bilirubin, one of the pigments of the bile.* Urobilin is more readily obtained from highly colored urines, (fevers, etc.): __ *A small amount of an alcoholic extract of the feces added to the urine will usually give favorable tests for urobilin. 32 Test. The ordinary test with an alcoholic solution of zinc may be simplified in the following manner: 10 cc. of urine are acidified with 2 drops of HCl and shaken with 2 cc. of chloroform. After the separation of the liquids, 2 cc. of the chloroform layer are tested with 4 cc. of a solution of 1 gram of crystallized acetate of zinc in a liter of 95% alcohol (shelf reagent). At the junction of the two layers the green fluorescent ring characteristic of urobilin will appear, and on shaking, a fluorescence which is rose-colored by reflected light will be distinguished throughout the liquid. Another test is to add ammonia to the urine until dis- tinctly alkaline, filter, and to the filtrate add a little 10% chloride of zinc solution.. A green fluorescence should appear, and if examined with the spectroscope, a characteristic band should occur. (See Fig. 16.) Leucin and Tyrosin are patholovic constituents of urine. They are normal products of pancreatic digestion and under ordinary conditions are carried, after absorption, to the liver, where they disappear, presently undergoing decomposition. When present in the urine these bodies are usually considered pathognomonic of acute yellow atrophy of the liver, although they are likewise stated to be present in the urine in certain rare cases of acute phosphorus poisoning associated with hepatic atrophy, due to typhoid fever, etc. Phenol. According to Tereg and Munk the horse excretes in the urine about 10 grams of tribromphenol in 24 hours. The tribromphenol is equivalent to 3 grams of phenol daily. Great importance is laid by these observers on the excretion of phenol, a process which is suspended during intestinal complaints, par- ticularly colic, and is, according to them and others, a cause of the rapid death in these effections, produced by the toxic effect of the unexcreted phenol. The production of phenol in the healthy body is greatly influenced by diet, being largest on rye and hay, one part peas and two parts oats, and on hay alone; it is smallest on rye alone, and next smallest on oats and hay. Sal- kowski is inclined to regard the excretion of 3 grams of phenol daily as too high. 33 We ABNORMAL SUBSTANCES FOUND IN THE URINE. Albumin. The presence of this substance in the urine is regarded as pathologic. There is, however, in the urine of some individuals apparently enjoying perfect health, minute traces of albumin sometimes present, and, unless these traces persist, are . not to be regarded as serious. If present in any considerable quantity, it must be regarded as distinctly abnormal. Albumi- nuria is the term applied when albumin occurs in notable quantity in the urine. The principal form of albumin present is serum- albumin, in addition there may be serum-globulin, acid albumin, albumose, and peptone. The amount of albumin in the urine may be increased: 1. By food rich in albumin; 2. Suppression of cutaneous perspira- tion, as by colds, burns, or cutaneous diseases; 3. Pulmonary and cardiac diseases attended with dyspnoea, cyanosis, valvular dis- eases of the heart; 4. Febrile and inflammatory diseases, as ma- larial, eruptive, typhus and typhoid fevers, croup, diptheria, erysipelas, rheumatism, gout, peritonitis, meningitis, etc.; 5. By lesions or prostration of the nervous system, especially when at- tended by diminished temperature and arterial tension, as from grief, fear, injury, pressure; 6. Pressure as from tumors, preg- nancy, etc.; 7. Cachexias, as from cancer, syphilis, scrofula, sep- ticemia; 8. Hydremia and ailments that disturb the vascular ten- sion; 9. Chorea, convulsions, exacerbations of febrile and other diseases; 10. Diseases of genito-urinary organs, as Bright’s dis- ease, cystitis, hemorrhage, abscess, etc.; 11. Medicines, such as copaiba, cubebs, turpentine, some emetics and drastic cathartics, some anesthetics, coffee, many metallic salts, poisoning by hydro- gen arsenide, carbon protoxide, carbon dioxide, phosphorus, iodine, iodoform, etc. The loss of organic material (albumin) disturbs nutrition, the blood becomes more aqueous; it sometimes produces an anas- arca which is the result of hydremia and of anuria. Albuminuria may be caused: 1. By an alteration in the renal transudation; 2. By certain changes in the blood; 3. By disturb- ance of the circulation. 34 Renal Changes. Lesions of the dialysing portions of the kidney, especially of the glomerule; the epithelium of the convo- luted tubules may also be essential in the production of albu- minuria. These cells normally prevent the albumin from filtering through with the other elements of the plasma. Albuminuria is also dependent upon renal lesions, sometimes primary as in nephritis, sometimes consecutive as in alteration in the blood or a disturbance in the circulation. Acute nephritis, chronic neph- ritis, fatty or amyloid degeneration interfere with the process of dialysis. Bacteria may exercise a traumatic action upon the renal epithelium and cause desquamation or degeneration, obstruct the vessels, modify blood pressure, or by the excretion of soluble products irritate the parts. Changes in the Blood. The dialysing membrane cannot stand with impunity any adulteration of the blood. The passage, in the kidney, of any such substance as biliary pigment in icterus; glucose in diabetes; poisons, such as alcohol, lead or mercury, or toxic gases, render the urine albuminous. Subcutaneous injections of solutions of extractives (leucin, tyrosin, creatinin, xanthin and hypoxanthin) cause degeneration of the epithelium of the kidneys and albuminuria. The subcu- taneous injection of tincture of cantharides causes, in a few min- utes, the production of an albuminous exudate in the glomerules. Although the limit of saturation of the blood plasma by albumin may be unknown, it is none the less evident that a superabun- dance of the substance (albumin) in the vessels causes albuminuria. The existence of a physiologic albuminuria is still doubtful in the domestic animals; for Fréhner, who has examined the urine of a number of healthy horses, has found only two cases in which albumin was present. In man it has been demonstrated to be due to severe muscular exercise, slight cold, and nitrogenous diet. Disturbance of Circulation. Too great a variation in blood pressure will cause albuminuria. Renal emboli, section of vaso-motor nerves of the kidney, medullary lesions, etc., cause albuminuria by causing an active congestion of the kidney. Venous stasis, organic affection of the heart, of the liver, presence of fetus, etc., cause albuminous urine. The urine may be less fluid. Bacteria may frequently cause 30 thromboses, emboli, edemas, anemias, ete., from vaso-motor trou- bles changing simultaneously the filter, the liquids which filter, with regard to pressure and velocity. Albuminous urine is ‘usually of light color and low specific gravity. It may occasionally be dark and dense, due to other ingredients, or to concentration. Under particular conditions of fatigue or disease albumin may appear in the urine. Temporary albuminuria is sometimes induced by a cold bath, especially in persons prone to kidney disease, and it has been observed after excessive muscular exercise, as in the urine of soldiers after a prolonged march. Any cause which leads to an increased blood pressure in the kidneys tends to induce albuminuria, and many of the cases in which it is the result of disease may be traced to this cause. Al- buminuria is a constant accompaniment of the nephritis follow- - ing scarlet fever and may occur to a less extent in pneumonia, typhoid and diptheria. It may also occur in diabetes, and is then a highly unfavorable symptom. In every case the urine must be clear before testing, by filtering it carefully; also take the specific gravity.* In addi- tion to the suspected urines make control tests on the normal for comparison. Heat Test. Heat about 5 cc. of the urine to the boil- ing point in a test tube. Note the slightest turbidity. If present it will be due to albumin or earthy phosphates. In horse urine the precipitate may be due to driving off CO, and precipitation of lime, etc., not phosphates. Add slowly a few drops of acetic acid (or nitric). If due to the phos- phates the urine becomes clear, if the turbidity remains it is albumin. Care must be taken, in the addition of the acid after boiling, to note the effect after each drop is added and to go on adding until there is no doubt that the urine is distinctly acid. If only a trace of albumin is present and too much acid is added the albumin may be converted into acid albumin and remain in solution. Heat does not coagulate acid albumin. Add a little acetic acid to dissolve any phosphates and heat again. *If a pathological urine is not available, a little blood serum added to the normal urine will give satisfactory tests. 36 Another Heat Test. Fill a test tube about one-third full of water and boil it. Add a few drops of the suspected urine. If albumin is present a cloudiness or coagulum will appear, according to the amount of albumin present. Heller’s Cold Nitric Acid Test. Pour some of the urine gently upon the surface of some nitric acid in a test tube. A ring of white coagulum occurs at the junction of the two fluids. If the quantity of albumin is small, the coagulum may not occur for a few minutes. A brown zone will frequently be seen at the point of contact due to the action of the acid upon the coloring matters of the urine, but it does not give any turbidity unless albumin be present. Millard-Robert’s or Nitric Magnesian Test. The re- agent is as follows: Nitric Acid, 1 part; Sat. Sol. Mag- nesium Sulphate, 5 parts. Use as in the preceding test. Picric Acid Test. (Johnson’s). Fill the test tube half full of urine. Slightly incline the tube and gently pour down its side about 2 cc. of a saturated solution of picric acid, so that it may come in contact with the upper layer of the urine. Place the tube in an upright position. A layer of coagulated albumin will appear at the line of junction. The coagulation of albumin takes place at once and is thus not easily mistaken for precipitated urates, which require some time for their precipitation and dis- appear on the application of heat. Ferrocyanide Test: To 2 cc. of acetic acid in a test tube add 4 cc. of a 5% solution of potassium ferrocyanide. Mix them and add 10 cc. of urine. Piate II. Various forms of Calcium Carbonate Crystals. (Horse Urine) . 7 * 2 w ‘ . ’ f _ 2% . - > s 61 posed of the acid urate of soda. austic potash does not dissolve it, hydrochloric acid gives a crystalline precipitate, the deposit will give the murexide test. (Page 26). Uric Acid. If a crystalline deposit, almost insoluble in boiling water, but soluble in caustic potash and giving the murexide test, it is uric acid. Cystine (very rare). The deposit of crystalline, insoluble in caustic potash but soluble in ammonia and hydrochloric acid. Pus. A white, dense, mucus deposit, viscid and not often mixing with the urine; caustic potash renders it more viscid; the microscope shows the pus corpuscles. This deposit is rare in acid urines. Blood. Red deposit; the tincture of guaiac with hydrogen dioxide gives a blue color. The spectroscope gives the charac- teristic lines in the spectrum. The hemin test will show the characteristic crystals. When blood is present the urine is gener- ally albuminous. Some deposits not very abundant and having no special chemical reaction may be recognized under the microscope. Neutral or Alkaline Urine. Alkaline Urates. More or less reddish deposits easily soluble in boiling water and giving the murexide test. Triple Phosphate or Calcium Phosphate. A white deposit, “UT A | cae ct ba “ORANGE YELLOW! GREEN Blue HOLE: Fic. 16 A.—Spectrum of Oxy-hemoglobin. B.—Spectrum of Reduced Hemo- globin. C.—Spectrum of Urobilin. 62 insoluble in boiling water, soluble in acetic acid, does not give the murexide test; acidified with nitric acid, the molybdate of ammonia solution gives a yellowish precipitate in the cold. Calcium Oxalate. White deposit insoluble in boiling water, also in caustic potash and acetic acid. Soluble in hydrochloric acid or in nitric acid; does not give the murexide test. Calc'um Carbona’e. White deposit insoluble in boiling water or in caustic potash. Soluble with effervescence in acetic, nitric or hydrochloric acids, does not give murexide reaction. Pus, (more frequent in alkaline and albuminous urine). A white, dense, mucus-like deposit, not mixing readily with the rest of the urine. Caustic potash renders it more viscid, the micro- scope shows pus globules. Blood. Determined by the guaiacum test, spectroscope, microscope, or hemin test. From the pathologic point of view the presence of urinary sediments generally indicate an alteration of the secretions. .@ Microscopical Examination of Urine. If an immediate examination is desired the centrifuge may be used to cause the sediment to separate from the urine; otherwise the urine is set aside for some hours in a cool place when the sediment gradually settles to the bottom. The supernatant liquid is poured off and by means of a pipette, camel’s hair brush or a wire loop a small portion of the deposit is transferred to a slide and Fic. 17. Uric Acid Crystals. ‘ examined. The deposits may be divided into unorganized and organized sediments. 7 % re YS Pa ‘, * > aC eee ee ee 65 Unorganized Sediments. In unorganized sediments two con- ditions may be considered: Ist, the urine is acid. 2nd, the urine is alkaline. In acid urine look first for crystals of uric acid diverse in form and reddish brown in color. (Fig. 17). Second, crystals of acid urate of soda, yellowish or red. The crystals are badly formed. Third, crystals of oxalate of lime also found in alkaline urine. (Fig. 18). Fourth, crystals of hippuric acid. Fifth, crystals of calcium sulphate. (4 and 5 are met with rarely in acid urine). Sixth, calcium phos- phate. Make sure of the identifi- cation by reference to charts. In alkaline urine look first for ammonio-magnesium phosphates, (Fig. 19), (triple | phosphates). Second, bicalcium phosphate crys- tals. Third, tricalcium or amorph- ous phosphate crystals. Fourth, crystals of sulphate of lime. Fifth, crystals of oxalate of lime, (also Fic. 18. Calcium Oxalate. found in acid urine). Sixth, crystals of urate of ammonium in the form of yellow colored spheres. Crystals of cystin, leucin and tyrosin are sometimes en- countered in acid or alkaline urines. ». BOrganized Sediments. These are brought more plainly into Fic. 19. Triple Phosphates. . tao" ¥ Mis + tia Ty hat We 64 view if the preparations are stained, although this is not uni- versally practised. Organized sediments may be divided into (a) histologic elements, (b) microbic elements. , Of the histologic elements there are frequently encountered, Ist, epithelial cells from the bladder, from the vagina and from the ureter, their presence has no special significance. If, how- ever, they are present in considerable quantity, a lesion of these parts may be suspected. 2nd, cells from the pelvis of the kidney, generally an indication of renal affection. 3rd, cancer cells, which have a special significance. 4th, hyaline or granular casts, seen more easily in stained preparations; they are encountered frequently in albuminous urine; their presence indicates a mild form of nephritis. 5th, epithelial hem- orrhagic or wax casts, of which the hemorrhagic are the most frequent, they are markedly colored and easily recog- nized on account of the hemoglobin they contain; they contain fine granulations which are not blood corpuscles but possible fragments of them, they indicate a severe form of nephritis. 6th, cylindroid ele- ments drawn out in an irregular and somewhat ribbon-like form; they are the product of the secretion of the urinary epithelium. In the human race they may be found in cases of scarlatina and generally in certain forms of nephritis. In examining for casts it is desirable to examine the urine immediately after emission, as they generally disintegrate rapidly and disappear. 7th, blood — corpuscles more or less cre- nated but easily recognizable by their yellowish tint; the ‘presence of blood corpuscles in the urine indi- cates a hemorrhage either in the blad- der orinthe kidney. 8th, pus corpuscles which may have crenated borders, granular contents and appear quite refractive, a drop of Fic. 20 Micrococcus Ureae. at 69 dilute acetic acid will render the nuclei visible. Pathologically the pus corpuscles indicate a suppuration of some portion of the urinary tract. In the majority of cases it is impossible to say if the pus comes from the bladder or kidney. In cases of cystitis of the neck of the bladder, the last portion of the urine passed contains no pus, while if this trouble be in the kidney this last portion will contain pus. This fact is of some use in diagnosing cystitis of the neck of the bladder. 9th, spermatozoa may be present but are easily recognized by their elongated form. 10th, mucus may be fre- quently present, but has no very great pathologic signi- ficance. Glycogen Cells. If the urinary sediment is stained with dilute Lugol’s Solution, there will be seen, in many specimens of % human urine, a num- “et ey re ber of epithelial cells Fic. 22. Red blood corpuscles crenated. present with more or less of their interior stained a deeper brown like glycogen, than in other cells that may be present. As yet, nothing has been determined in regard to their importance. Amyloid or Amylaceous Bodies. These are small circular or oval bodies which give the starch reaction with an iodine solu- tion. Just what their significance is, is not known. Virchow first described them. They have been found in various tissues and excretions, including the urine normal as well as pathological. In diseases of the kidneys, Veitz and Wederhake found that these amyloids afford us important indications; but that in affections of the bladder they are present in increased quantity. Weder- hake therefore suggests that the amyloid bodies have a ‘certain value in differential diagnosis. The absence of the bodies in pathological urine is against catarrhal disease of the bladder; whilst their presence, especially if numerous, in urine which + fm . - - x * « - — we i i he ‘= : ~ cm ll i 5 go Z ie . — 4 . c ’ . ‘ » . , ’ * G ; . é 7 FE; 7 er é ; — an ‘ oer oe ; “s st ee eS a oot Reo tye pe etine pN reheat re ee “a Re. Ob ae ele 7 —.. iG + yn =>, ? wh = hy i . in ie ae a “s — + = 23 = ts > . a ~ ‘ ~ at 67 appears to contain renal elements only, indicates that the bladder is also affected. (Dixon Mann). The urinary sediment stained with a dilute Lugol’s solution show these bodies, when present, varying in color from a blue to a deep blue black color. Urinary Casts. Although probably observed earlier, Henle is credited in 1842, with first carefully describing the casts moulded in the tubules of the kidneys. Among earlier views, casts were considered as being composed of coagulated fibrin; as pro- ducts of the secretion of the epithelium of the tubules; as trans- formed or disintegrated epithelial cells and as products from the blood. A view quite commonly accepted is that casts are the products of the coagulation of albuminous material. ‘The fact that the presence of casts in the urine is usually accompanied by the presence of albumin lends force to this view; for the more abundant the albumin, the more likelihood is there of finding casts. From this standpoint, then, casts, may be regarded as albuminous exudates from the blood, with the addition of transformed or de- stroyed epithelium. In nearly all cases where casts are present, some — albumin is found. Occasion- ally they are sometimes described as being present without the ap- pearance of albumin. This condi- Fic. 23. a. Showing forma- tion is looked upon with some tion of hyaline cast in tubule. doubt; for it is commonly be- b. Hyaline cast with granular lieved that albumin exists but in °POSit © Granular cast. an amount too slight to be detected by the ordinary chemical tests. The presence of casts in the urine is of much diagnostic im- portance; if found in any quantity, they indicate nephritis par- ticularly if albumin is also present in any amount. It is claimed by some that merely hyperemia of the kidney will cause the appearance of casts in the urine, and that they may sometimes be found when the kidneys are perfectly intact. Mitchell states that two or three hyaline or one or two small granular casts may a a i ce 68 be found in one of every three specimens of the twenty-four hour’s urine examined (human). Casts have been described in cases of gastro-intestinal catarrh, in jaundice, in acute and chronic anemia, as well as in nervous affections of different kinds, with- out accompanying inflammation of the kidneys. There are many, however, who hold that casts are always the products of an inflammatory process, and are, therefore, indicative of renal inflammation. Other evidences or symptoms should also be taken into account. Occasionally it is difficult to find casts even when t Hew ane known to exist. Alka- line urine has a tendency to dissolve them. At. 1 ives they will not settleiomg hours. Usu- ally if al- lowed to Ss tanith. Sie hours, the casts will set- tle if they are present. The centrifuge will — bring them down in'a few minutes. A low power of the micro- scope (150-200 diameters) may be used in the search for casts and will enable the observer to pass over the field quite rapidly. To identify the cast and its structure, a power of 400-500 diame- ters is required. More thanonespecimen must always be examin- ed before giving a positive statement as to the presence or ‘ absence of casts. False or pseudo-casts have been described; these are believed to be accidental formations, while true casts in general indicate nephritis. Fic. 24. a. Blood cast and hyaline cast carrying blood cells. b. Pus cast. c. Hyaline cast carrying epi- thelial cells. d. Epithelial casts. (Greene). 69 True casts may appear in three different sizes according to - the portion of the tubule in which they are formed. The smaller size originates in the narrow tubule. The next in size comes from the convoluted tubule of the second order (no casts from the convoluted tubule of the first order, i. e., that portion of the tubule nearest the glomerules, appear in the urine, because they are too large to pass through the narrow tubules). The third and largest are those developing in the straight collecting tubules. It is believed a prognostic value may be attached to the size of the casts as well as to their number. Casts from the narrow tubules indicate a mild attack; from the convoluted tubules a severe form, especially in the cortex of the kidney; from the collecting tubules, with the other forms also present, a serious condition of general renal inflammation or unfavorable prognosis. The identity of casts may generally be determined by their uni- form width. They are usually longer than they are broad, and have one well- rounded ex- tremity and well defined borders. True casts are of six va- rieties; hya- line, epitheli- Cabesb lo OG granular, fat- ty and waxy cases...” In” /a general way the first three varities are found in an acute form of nephritis. Duwran'g the first few weeks of the inflammation the last three varieties are not encountered. Ifthe acute condition passes into a subacute, then the granular variety appears, at first in small number, then in larger, with, usually, a considerable number of the hyaline and Fic. 25. Granular Casts. (Greene). 70 epithelial casts. Fatty and waxy casts are always secondary products, and as a rule not found until a nephritis has existed for some time. Hyaline casts are colorless, pale, more or less transparent formations soluble in acetic acid. They are of variable size and generally difficult to detect on account of their apparently struc- tureless character. At times a slight granulation may be seen imbedded in or adhering to their matrix and occasionally acci- dental attachments of pus or fat globules in small numbers. Epithelial casts have a hyaline ma- trix more. of, less concealed by epi- thelial cells. The presence of these casts is indicative of an acute process. Blood casts con- sist of the hyaline matrix with blood corpuscles imbed- ded in or adhering to the matrix. Pus casts are rare, but when present the Fig. 26. a. Fatty casts. b. and c. Blood casts. d. Free fatty Molecules. (Roberts). pus corpuscles ad- here to the matrix. Blood casts are indicative of a hemorrhage into:the tubules and of an acute hemorrhagic process. Hyaline and epithelial casts are usually associated with them. Granular casts usually have well defined boundaries with granular matter imbed- ded in or adhering to the matrix. They may be finely or coarsely granular, the lat- ter having a more serious significance. Granular casts are due to a disintegration of the renal epithelium. Their py¢. 27. Fatty Casts and Fat Droplets. 71 degree of refraction is changeable; sometimes they appear yel- lowish, at other times colorless. Fatty casts have a hyaline matrix containing a number of small, glistening fat globules and granules. Some free fat is also usually found in the field. As fatty casts are secondary products of epithelial and granular casts, the diagnosis of a chronic process is justifiable. The hyaline matrix is character- istic of the different varie- ties of casts that have been mentioned up to this point. Waxy casts differ in chemical composition from those previously men- tioned. They are charac- terized by wavy contours; a high refracting power; a more or less yellowish color and quite a high degree of brittleness. They are slowly, if at all, attacked by acetic acid. Their presence signifies waxy degeneration of the kidney. Hyaline casts may sometimes have a superficial resemblance to waxy casts, but ey never have the same high refraction as the latter. In the urine of the horse, the sediment of crystals of lime car- bonate may obscure the search for casts. In this case add a few drops of acetic acid which will dissolve the crystals quickly, and the casts, if present, will show more distinctly but undergo solution more slowly than the crystals. Cast-like formations are composed of various elements having a form somewhat similar to casts but lacking the matrix soluble in acetic acid. Amorphous urates often simulate granular casts in form. Bacteria are often grouped in a manner similar to the form of cast, but a close inspection shows an irregular outline, and usually a number of groupings not in cast form. Granular detritus and hematoidin may also assume the form of casts; like- wise epithelial cells, blood corpuscles and fibrin in renal hemor- rhages may also assume the form of casts. Acetic acid is said to be a reliable reagent for differentiating between true and false Fic. 28. Waxy Casts. ow oJ Peay 7 o : t . : . i Ke L heh : ae oa “> 7 a i - f j ¥ uY- ‘ #2 Be | ' _ , 4 * : oa ats * ; ’ 4 : Se APR FT ¢ i, + pe A ; : Se a . vk i fe \ » fi & m : eh. 4 FAR vhs date ; H REE eee jag 4 BS : (7S 2 oe t ; . ta og jae ; | ; ' ; ; 7 Fé Piet al wt cee Tea é | # -s “ et ees Fete) FC | > , Ma ane ay Ai, aMos e 4 fe Mae stthate: a Nr ; iene ar ¥ ~] bo casts. This reagent dissolves the matrix of the true casts but does not act upon the cast-like formations. . _ Cylindroids appear like hyaline casts, but are large and band-like. Their breadth is uniform and they often contain Fic. 29. Cylindroids. a. b, Cast-like forms; cc. Filamentous forms. (Ogden). crystals, epithelial cells and corpuscles. They are soluble in acetic acid and are of renal origin. No special sig- nificance is attached to them. Mucous cylinders sometimes call- ed cylindroids are usually not of uniform breadth and seldom contain morphologic constituents and are in- soluble in acetic acid. They are usu- ally found in any urine containing an abundance of mucus and are of no special significance. Upon special blanks, test specimens of urine quantitatively, the number and amount of constituents being unknown. ~~] we) Form used in examination of Horse Urine. Sample Owneriiccain SNe. 22. eeeeeddrecs Now. 0) 3, Sospebies fey) Siekis , sAweiis ee Sexi Object ats... os. eee. Date a. 22 ee Normal (Horse). Amount in 24 hours 3000-4000 ec. c. Specific Gravity 1025-1050 Reaction Alkaline Color Yellowish brown Translucency Turbid Consistency Viscid Total Solids 50-120 parts per 1000 Chlorides 8-14.“ Sulphates 2-3 ie az Phosphates 05=7ates ce Urea 20-40 * a Uric Acid Trace Hippuric Acid 4— 8 parts per 1000 Indican i ey ae: ss ABNORMAL CONSTITUENTS. Albumin Sugar Bile —~ Hemoglobin MICROSCOPIC EXAMINATION. Epithelial Cells Leucocytes Blood Casts Spermatozoa Micro-crganisms Crystalline Deposit. Calcium Carbonate Calcium Oxalate Triple Phosphates 74 Form used in Examination of Human Urine. Name . AG@reES si Ai. Dolce SER, tai le een ee Date. Normal. Sample. Amount in 24 hours. 1000-1500 cc. Specific gravity 1015-1025 Reaction Acid (30°—40°) Color Light golden Translucency Clear Sol ds 34-50 parts per 1000 90-75 parts per 24 hours Chlorides 6-9 parts per 1000 as NaCl 10-15 parts per 24 hours 1.5-2.5 parts per 1000 as P.O; Phosphates 2.—3.5 parts per 24 hours 1.5-3 parts per 1000 as SO* Sulphates 2.5-4 parts per 24 hours Urea 14-22 parts per 1000 22-33 parts per 24 hours Uric Acid 0.25-0.40 parts per 1000 0.40—0.60 parts per 24 hours Indican Normal, little, medium, much Albumin ‘ Sugar Bile Hemoglobin MICROSCOPIC EXAMINATION. Epithelial cells Leucocytes Blood Casts Crystalline Deposit. Uric Acid Oxalates Triple Phosphates Urates =~] Or Procedure in Examining a Sample of Urine. Experience has shown that the following procedure expedites — the work of examination. Pour the urine into a sedimenta- tion glass, as some little time may be required for the sediment to settle. After noting the amount, color and translucency, take the specific gravity with the urinometer. This may be done in the sedimentation glass. Filter as much of the urine as may be needed for the subsequent tests. While waiting for the urine to filter make the tests for urea and uric acid as the correct reading is not obtained until some few minutes after the tests have been made. Test the reaction with litmus paper or if the degree of acidity is required (human) use the acidi- meter. Test forindican. Thus far the tests may be made just as well with the unfiltered as the filtered urine. For the albumin and sugar tests the filtered urine must be used, using two or three different tests for each substance. If albumen is present it should be removed before making the sugar tests. Also before making the centrifuge tests for the chlorides, phosphates and sulphates. This may be done by adding a little acetic acid to the urine and boiling it until the albumin is coagulated. Filter out the albumin and use the urine thus filtered for the remaining tests. With a pipette remove some of the sediment at the bottom of the sedimentation glass and introduce it into a centrifuge tube filling the remainder of the tube to the required height with the unfiltered urine. Include this’ with the other centri- fuge tubes used in making the tests for the chlorides, etc., revolv- ing them in the centrifuge for three minutes. If albumin is present and it is desired to know the amount, it may be deter- mined by the centrifugal method as described in the text. For the microscopial examination, prepare a couple of slides from the sediment in the centrifuge tube, after pouring off the clear urine above. It is well, also, to prepare a slide’ or two from the sediment in the sedimentation glass. After putting a drop or two of the sediment on the slide it may be covered with- a cover glass and examined clear or before’ covering the sedi- ment a drop of dilute Lugol’s solution, or safranin or other stain may be added. In some cases the slight tint of the dye appears to make the casts or other elements stand out more clearly. To detect the amyloid bodies or glycogen cells it 1s necessary 76 to use the iodine solution and this often times serves to make the casts and other elements more distinctly visible. APPENDIX. Formulae for Reagents. Barium Chloride Solution for Centrifuge Sulphate Test. Barium chloride, 4 parts Hydrochloric acid, 1 part Distilled water, 16 parts Barium Chloride Solution for Quantitative Test of Sulphates. Pure crystallized barium chloride, 30.5 grams. Distilled water to 1000.-ser Baryta Mixture. This mixture is prepared by making sat- urated solutions in the cold, of barium nitrate and barium hydrate, and adding two volumes of the hydrate to one volume of the nitrate. Benedict’s Solution. Solution 1. Cupric Sulphate 34.65 grams Distilled water up to 500. cc. Solution 2. Sodium carbonate (anhy-* drous), 100. grams Rochelle salt, 173. grams Distilled water up to HOO! Mec Benedict’s Single Solution. Cupric Sulphate, 17.3 grams Sodium Citrate, 173. grams Sodium Carbonate (anhydrous), 100. grams Distilled water up to 1600: sce: Benedict's Solution for Quantitative Tests. Copper Sulphate Crystals, 18 grams Sodium Carbonate Crystals, 200 grams Sodium Citrate, 200 grams Potassium Thiocyanate, 125 grams Potassium Ferrocyanide (5 %Sol.) 5 ce. Distilled water to 1000 cc. Dissolve the citrate, carbonate and thiocyanate in 800 cc. ~ SY of distilled water with the aid of heat. Dissolve the copper sulphate separately in 100 cc. of distilled water and mix the two solutions slowly with constant stirring; then add the ferrocyanide solution. Cool and dilute with distilled water to exactly 1000 ce. Cochineal Solution. Boil 40 grams of cochineal in 800 cc. of water. When cool add 200 cc. of alcohol and filter. Decinormal Oxalic Solution. Dissolve 6.285 grams of pure oxalic acid in enough distilled water to make at or near 15° C. (59° F.), exactly 1000 cc. Decinormal Sodium Hydroxide Solution. Dissolve. 3.996 grams of pure sodium hydroxide in enough distilled water to make, at or near 15° C. (59° F.), exactly 1000 cc. Ehrlich’s Diazo-reaction. Solution 1. Sulphanilic acid, 2 grams Hydrochloric acid, 50° ec: Distilled water, 1000 cc. Solution 2. A 0.5% solution of sodium nitrite. Use in the proportion of 1 part of No. 2 to 50 parts of No. 1. Esbach’s Reagent. Picric acid, 10 grams Citric acid, 20 grams Distilled water up to 1000 cc Fehling’s Solution. Solution A. Pure Copper Sulphate crystalline, 34.64 grams Distilled water up to 500. “ee: Solution B. Sodio-potassium tartrate (Rochelle salt), 173. grams Pure caustic potash, 125. grams Distilled water up to 500: eee Use equal parts of A and B. Hypobromite Solution. Dissolve 40 grams of caustic soda in 100 cc. of distilled water. To 23 cc. of this solution add 2 cc. of bromine. To this mixture add an equal volume (25 cc.) of water. The solution is not very stable and is best made up as needed. Lugol’s Solution. Iodine, 2.5 grams Potassium iodide, 5. grams Distilled water to 50. cc. This may ordinarily be diluted 1 to 5 or 10 for urine work. Magnesia Mixture. Magnesium sulphate, 1 part. Ammonium chloride, 1 part Ammonia water, ; 1 part Distilled water, 8 parts Millard-Roberts Reagent. Nitric acid, 1 part Sat. Sol. Magnesium Sulphate, 5 parts Mohr’s Volumetric Method for Chlorides. Solution 1. Fused silver nitrate, 29.075 grams Distilled water to make, 1000. Ce; Solution 2. Neutral potassium chromate, 10. grams Distilled water to make, 100... “ee: Nylander’s Reagent. Digest 2 grams of bismuth subnitrate and 4 grams of Rochelle salt in 100 ce. of a 10% solution of potas- sium hydroxide. The reagent should then be cooled and filtered. Obermayer’s Reagent. Dissolve 2 grams of solid ferric chloride in 500 cc. of concentrated hydrochloric acid. Sp. gr. 1.19. Phenolphthalein. Dissolve 1 gram of phenolphthalein in 100 cc. of 95% alcohol. Ruhemann’s Uric Acid Reagent. Iodine, 0. 5 grarns Potassium iodide, 1.25 grams Absolute alcohol, 7.5 grams Glycerine, 5. grams Distilled water to 100. grams Salicylic Adehyde. Dissolve 1 gram of salicylous acid in 10 cc. of absolute alcohol. Shieb’s Reagent. Solution 1. Ammonium Sulphate (purest), 1.2 grams Copper Sulphate (purest), 2.6 grams Distilled water up to HOS) eG ae 79 Solution 2. Caustic potash C. P. 20. grams 7): Distilled water up to ~ 50. ce. re Dissolve and when cool, add Glycerine, DORACC. Ammonia water, 0.960 sp. gr. 300. cc. _ Add No. 1 to No. 2 and dilute the whole.to 500 cc. with distilled water. Stopper securely and shake until thoroughly mixed. | " aes - 4