Urine Investigation

Urine analysis A routine urine analysis consists in tree parts: the physical, microscopic, and chemical examinations.

Physical Examination of Urine The physical examination of urine includes observing the color and appearance, and measuring the specific gravity. It is the easier and the quickest urine analysis and it can be performed while the urine specimen is being prepared for other procedures. Physical characteristics are the best observed immediately after voiding, before the sample is refrigerated, refrigerated specimens should be allowed to rich room temperature before the analysis is performed. Urine odor normally recently voided urine has a characteristic aromatic and not unpleasant odor. Changes in the urine odor may be due to dieses, diet, or the presence of microorganisms. In cystitis, or other urinary tract infections, or if urine is allowed to remain unrefrigerated for a few hours, bacteria may break down urea to form ammonia; the resulting odor is similar to ammonia. The fruity odor is because the presence of ketones (uncontrolled diabetes of ketosis in cattle or sheep). Urine color the normal color ranges from pale yellow, to straw, to amber. Dilute urine samples are pale; more concentrated urine samples are darker yellow or amber. Variation in color may be cause by diet, medications, physical activity, and diseases. Red urine, if it is cloudy, may be due to hematuria (the present of erythrocytes), while clear red urine may be due to the presence of hemoglobin (hemoglobinuria), brown urine occurred in myoglobinuria (the presence of myoglobin that follow to rhabdomyolysis), in this case the urine turns to brown color. Yellow brown or green brown urine is usually found in bilirubinuria (hepatic and posthepatic jaundice). Urine appearance usually urine is clear (not in the horses that normally show opalescent urine due to increased precipitated salts). The cause of cloudy or turbid urine specimen becomes evident during the microscopic examination. Fresh normal urine is clear immediately after voiding, at room temperature, or at 4C it may became turbid or cloudy (depending of the pH the cloudiness my be due to urates of phosphates). In freshly voided urine turbidity or cloudiness may be because WBCs, epithelial cells, mucus and bacteria. Specific gravity 1.005-1.030, with most samples falling between 1.010 1.025 indicated the concentrations of substances as urea, phosphates, chlorides, proteins, and sugars.

Microscopic Examination of Urine Microscopic examination of the urine sediment may reveal infections, trauma o urinary tract, the presence of abnormal crystals (suggest a metabolic disorder). All urine specimens should be examined as soon as possible to avoid cellular deterioration or multiplication of the bacteria. 10 ml of well mixed urine is centrifuged at 1500 2000 rpm (4000xg) no more, for 5 minutes. The supernatant is carefully poured off, leaving 0.5 ml in the tube, than is resuspended by gently tapping the tip of the tube. One drop of mixture is placed on the slide and covered by coverglass. Urine sediment refers to the solids that settle to the bottom of the urine specimen after centrifugation, or when urine is allowed to stand undisturbed. It may contain cells, microorganism, crystals, casts, amorphous material. Urine sediment may be observed unstained or colored to methylene blue. Cells in Urine Sediment (high power field Ob - 40x) RBCs Size: 7-8 m in diameter, pale or yellowish, smooth biconcave disk, no nucleus or cytoplasmic granules. Normal: Less than 2 RBC/HPF (in females consider the genital tract), larger numbers hematuria. WBCs usually neutrophils, larger than RBC, 10-12 m in diameter, contain nucleus and cytoplasmic granules. Normal: no more than 2 RBC/HPF. Can originate from any part of the urinary tract (glomerulus to urethra) large numbers In inflammatory processes of the urinary tract - pyuria. Epithelial cells are large and flat, with distinct nuclei and much cytoplasm, the most common are squamos epithelial cells. Bladder and tubular cells are smaller. Renal Tubular Epithelial Cells are slightly larger than WBC, flat, cuboidal or columnar with one large round nucleus. They occur in tubular damage, pyelonephritis, ATN, salicylate intoxication, transplant rejection. Transitional Epithelial Cells are 2-4 times larger than leukocytes, round, pear-shaped, tail-like projections with large round nucleus. Squamous Epithelial Cells are large, flat, irregular-shaped cells originated principally from the urethra and vagina.

Urine Investigation Casts in Urine Sediment (high power field Ob - 40x) Hyaline Casts are most frequently observed, they are colorless, homogenous and transparent with rounded ends. This casts can be found in normal urine, following physical exercise, and dehydration. Waxy Casts are very high refractive index, yellow, gray or colorless, smooth homogeneous appearance, short, broad with blunt or broken ends cracked or serrated edges. They are found in severe chronic renal failure, malignant hypertension, and diabetic nephropathy. Granular Casts (and fine granular casts) are the second-most common type of cast. They have large and coarse granules, and fine granular casts show fine granules, gray or pale yellow. Granular and fine granular casts can result either from the breakdown of cellular casts, or the inclusion of aggregates of plasma proteins (eg, albumin) or immunoglobulin light chains. They are found in chronic renal disease; and as with hyaline casts, can also be seen for a short time following strenuous exercise. Red Cell Casts are brown to tan. They occur in renal hematuria, glomerular disease (acute glomerulo nephritis, lupus nephritis, renal trauma). White Cell Casts are found in infection and noninfectious renal inflammation (ie, acute pyelonephritis, interstitial nephritis and lupus nephritis) Epithelial Cell Casts are found in stasis and desquamation of renal tubular epithelial cells following tubular damage and necrosis. Fatty Casts (Oval Fat Bodies) show a Maltese-cross pattern under polarized light. They are found in fatty degeneration of the tubular epithelium in degenerative tubular disease. Crystals and Amorphous Deposits in Urine Sediment in acid urine Uric Acid has many different shapes, diamond, rhombic prism or rosette, and yellow or brown color. They are very common and can be normal occurrence. They might be associated with increased purine metabolism. Calcium Oxalate are colorless, octahedral or envelope. They appear in ingestion of oxalate-rich foods: spinach, rhubarb, tomatoes, garlic, oranges, asparagus, high intake of ascorbic acid, and in ethylene glycol poisoning. Cystine are colorless, refractile, hexagonal plates. Of diagnostic importance they are in congenital cystinosis or cystinuria. Leucine are oily, highly refractile, yellow or brown spheroids and concentric striations. They are mainly occurred in severe liver disease. Cholesterol crystals are large, flat, transparent and with notched corners. They appear in excessive tissue breakdown, obstructed lymphatic flow, nephritis and nephrotic conditions. Tyrosine crystals are very fine, highly refractile needles, black, yellow, in sheaves or clusters. They occur in severe liver disease Sulfonamides are fan or sheaf of needles, eccentric binding, clear or brown. Crystals and Amorphous Deposits in Urine Sediment in alkaline urine Triple Phosphates (Ammonium Magnesium Phosphate) are colorless prisms, 3-6 sides, oblique ends, and have coffin lids shape. They can be found in normal urines, and in chronic urinary inflammation. Ammonium Biurates are yellow-brown spherical bodies with long, irregular spicules. Calcium Phosphate is long thin, colorless needles, with one pointed end, and arranged as rosettes or star. They can be found in normal urines Microorganisms Bacteria round or rod shape structures Yeasts smaller than RBCs, are ovoid, budding and chains the most common is Candida albicans . to make a distinguish between yeasts and RBCs one drop of acetic acid is added, RBCs will lyse while yeasts will not. Protozoa Trichomonas vaginalis (in cattle) Spermatozoa

Chemicall Examination of Urine Protein proteinuria may have two mechanisms. First, it indicates that the permeability the glomerulus is abnormally increased (albumins are the first proteins that pass throughout the filtrating membrane albuminuria can be an indicator of damage to the kidneys). Causes of albuminuria can be discriminated between by the

Urine Investigation amount of protein excreted, the larger amounts are excreted in the nephrotic syndrome, far less albumins are found in nephritic syndrome and in diabetic nephropathy. Secondly, it may be caused by renal infections, when proteins are in fact the desquamated endothelial and inflammatory cells.

Glucose glycosuria - normally, urine contains no glucose because the kidneys are able to reclaim all of the filtered glucose back into the bloodstream. Glycosuria is nearly always caused by elevated blood glucose levels (when the blood glucose level exceeds about 180 200 mg/dl), most commonly due to untreated diabetes mellitus. Sometimes, glycosuria is due to an intrinsic problem with glucose reabsorption within the kidneys themselves, a very rare condition termed renal glycosuria. Glycosuria leads to excessive water loss into the urine with resultant dehydration, a process called osmotic diuresis, which leads to polyuria. Ketone bodies ketonuria occur when there is carbohydrate deprivation, such as starvation or diabetes mellitus, when the body relies increasingly on the lipid degradation for energy, in the lack of available glucose. The metabolism of fat proceeds in a series of steps: triglycerides are hydrolyzed to fatty acids and glycerol; the fatty acids are hydrolyzed into smaller intermediate compounds (acetoacetic acid, betahydroxybutyric acid, and acetone - collectively known as ketone bodies). Finally, the intermediate products are utilized in aerobic cellular respiration; the process requires the presence of glucose. In the absence of glucose they accumulate and are responsible for ketosis, followed by metabolic acidosis, which may lead to coma. Part of them are eliminated through urine, acetone is removed by the lungs. Bilirubin bilirubinuria is the detection of conjugated bilirubin in the urine. The term biliuria refers to the presence of any bile pigment in the urine. The cause of bilirubinuria is hepatocellular and mechanic jaundice, in humans it is also found in some inherited conditions that affect bilirubin synthesis. Blood - hematuria is the presence as intact RBC which indicates bleeding or hemoglobinuria is the presence of hemoglobin (due to massive hemolysis). In this case RBCs are absent. Calcium hypercalciuria - is the condition of elevated calcium the urine. Chronic hypercalciuria may lead to impairment of renal function, nephrocalcinosis, and renal insufficiency. Phosphate - phosphaturia is the hyperexcretion of phosphate in the urine. This condition is divided into primary and secondary types. Primary, hypophosphatemia is characterized by direct excess excretion of phosphate by the kidneys, secondary causes, including all three types of hyperparathyroidism cause hyperexcretion of phosphate in the urine.