URINALYSIS

Urine formation
1. Glomerular filtration: passive process of filtration from plasma
a. Forces favoring glomerular filtration: hydrostatic pressure
b. Forces opposing glomerular filtration: colloidal osmotic pressure, renal intratubular pressure, selective permeability
of glomerular capillary wall
2. Tubular reabsorption: active process for things the body wants to keep
a. Water, Na+, Cl-, low mw proteins, glucose, amino acids, water soluble vitamins
b. When [blood] of a substance exceeds glomerular threshold, substance spills out into urine
3. Tubular secretion: active process = H+, urate, NH3, K+
4. Urine concentration: only 1% of glomerular filtrate is excreted as urine; rest is reabsorbed
a. Water and solutes are reabsorbed in proximal tubules
b. Countercurrent system  concentration gradient in medullary interstitium
c. ADH  distal tubules and collecting ducts permeable to water
d. Aldosterone  secretion of K+/H+ and absorption of Na+ and water in distal tubules
Indications for UA
1. Part of minimum database
2. Suspect urogenital disease
3. Screening for asymptomatic disease
4. Monitoring therapy

Methods of collection
Method Advantages Disadvantages Overall
Free catch Noninvasive Contamination Prefer midstream, morning
Can do at home OK for USG
Large animals Not ideal for culture
Catheterization Relieve obstruction Trauma Less contamination than free catch
May induceUTI
Difficult in females and cats
Cystocentesis Immediate sample Iatrogenic RBCs Sample of choice for culture
Best for culture Tear of bladder
Aspirate of other structures
Manual expression None Trauma to bladder Not recommended
Reflux of bacteria
Sample handling- SUBMIT 5 mls of urine in a syringe asap or refrigerate the sample. For bacterial culture, put approx 3 mls
in a culture vial.
1. Process immediately
2. If not possible within 30mins, refrigerate and do chemical evaluation ASAP
3. Keep away from light (degrades bilirubin)
4. Cells degrade and chemical parameters get altered on standing urine
5. Refrigerated urine OK for sediment, frozen urine OK for chemical
6. Generally refrigerated urine is OK for 6-8hrs, useless after 24hrs

COMPONENTS OF UA: all 3 must be performed for meaningful results

PHYSICAL EXAM
1. Color
a. Normal: light yellow, yellow, amber (urochromes)
i. Rabbits may have yellow, green or red from food pigments
b. Abnormal
i. Red to red-brown: hematuria, hemoglobinuria, myoglobinuria
ii. Dark brown to black: methemoglobinuria, melanin
iii. Yellow-brown: bilirubinuria
iv. Orange-yellow: bilirubinuria, highly concentrated urine, phenazopyridine
v. Blue: Pseudomonas infection
vi. Pink: porphyrinuria (seen under UV light)
2. Turbidity
c. Normal:
i. Dog/cat = clear (may become turbid on standing or refrigeration)
ii. Cat: may be turbid from lipid droplets
iii. Horses: turbid from calcium carbonate crystals and mucous
 iv. Cattle: clear but may be turbid on standing from precipitation of Ca carbonate crystals
v. Rabbits/guinea pigs: thik and white d/t Ca carbonate crystals
d. Increased: crystalluria, high cellularity, mucous, lipid, sperm
3. Odor
e. Normal: produced by volatile fatty acids; intensity varies among spp and sex
f. Abnormal
i. Ammonia: urease-producing bacteria (Proteus, Staph)
ii. Ketones: sweet or fruity odor
iii. Putrid: bacterial degradation
4. USG: solute concentration measured as (1) USG or (2) urine osmolality
g. USG = measured with refractometer; measures refractive index of light passing through fluid
h. Species-specific calibration; periodically calibrate with distilled water
i. Temp should be 60-100F
j. Increased: protein, glucose, contrast agents (correction factor not used)
k. Normal: 1.001-1.065 in most, up to 1.080 in cats; adequate = 1.030ish
i. Hypersthenuria (bariuria) = high SG
ii. Isosthenuria = SG equal to plasma (1.008-1.012)
iii. Hyposthenuria = SG lower than plasma (<1.008)
l. Know hydration status of patient
m. If no other factors interfering, urine should be maximally concentrated with 3% dehydration

Low USG (nonrenal) Mechanism

Drugs Various
Hyperadrenocortisim Cortisol interferes with response to ADH
Severe hepatic disease ↓urea production = medullary washout
↓metabolism of hormones = cortisol, aldosterone; hypokalemia, Psychogenic
polydipsia
Diabetes mellitus Osmotic diuresis-glucosuria
Central DI Absence or deficiency of ADH
Nephrogenic DI Failure of kidneys to respond to ADH
Hypercalcemia Induces insensitivity of collecting ducts to ADH
Pyometra/sepsis Toxins inhibit ADH activity
Hypoadrenocortisism Renal Na loss = medullary washout

CHEMICAL EXAM
1. pH: reagent pad has 2 pH indicators: methyl red, Acid pH Alkaline pH
bromthymol blue Meat based diet Vegetable based diet
a. Can detect pH 5-9 Acidifying diet Urinary tract infection
b. Normal dog/cat = 6.5-7.5; normal large animal Metabolic acidosis Post prandial
= 7.5-8.5 Respiratory acidosis Alkalinizing agents
2. Protein: normal is NEGATIVE for all species Protein catabolic state Metabolic alkalosis
a. Glomerular filtration of protein dependent on Paradoxical aciduria in Respiratory alkalosis
size, charge, shape metabolic alkalosis Artifact – air degradation
b. Dipstick: tests mainly albumin
i. False positive: alkaline urine, contamination with quaternary ammonium compounds (cleaning solutions)
c. Sulfosalacylic acid precipitation test (Bumin test): precipitation of protein  turbidity
i. Sensitive for more proteins: globulins, Bence Jones
ii. False positive: X ray contrast substances, penicillin, sulfas
iii. False negative: highly alkaline urine
d. Microalbuminuria test: can detect <30mg/dl for dog/cat; SPECIES-SPECIFIC tests
i. Quantitative: ELISA
ii. Semi-quantitative: USG adjusted to Proteinuria Mechanism
1.010 Hemorrhage Albumin and globulin from plasma
e. Interpret urine protein with USG: protein is UTI Plasma derived proteins
MORE significant when urine is DILUTE Renal disease Glomerular disease: GN, amyloid
f. Urine protein:creatinine ratio: perform Tubular disease
with inactive sediment and NO azotemia Pre renal Increased glomerular permeability
i. Normal: dogs <0.5, cats <0.2 Overwhelmed tubular resorption
ii. >1 = glomerular or tubular Alkalinity False positive with dipstick
iii. >5 = glomerular damage
3. Glucose: freely filtered at glomerulus, reabsorbed completely in proximal tubules
 a. Normal is NEGATIVE
b. Glucosuria when [serum] > resorptive capacity
c. Dipstick: peroxidase rxn = measuring metabolite of reaction
d. False positive: sodium hypochlorite, hydrogen peroxide
e. False negative: ascorbic acid in urine
Glucosuria + persistent Glucosuria + transient Glucosuria +
hyperglycemia hyperglycemia normoglycemia
Diabetes mellitus Stress Primary renal
Hyperadrenocorticism Catecholamines glucosuria
Hyperglucagonemia Drugs: steroids, ketamine, Fanconi syndome
Hyperpituitarism xylazine, Ethylene glycol
Acromegaly Acute pancreatitis
Hyperthyroidism
Acute severe colic Species Threshold
Bovine milk fever Dog >180 mg/dl
Moribund animals Cat > 280 mg/dl
Cow > 100 mg/dl
Horse/calf > 160 mg/dl
Bird > 600 mg/dl

4. Ketones: acetone, acetoacetic acid, B-hydroxybutyrate

a. Freely filtered by glomerulus, completely reabsorbed by proximal tubules
b. Ketonuria detectable prior to ketonemia
c. Ketonuria: carb depletion/lipolysis = DM, starvation, insulinoma, pregnancy ketosis
d. Measurement: reagent strip OR tablets; both use nitroprusside reaction
i. More specific for acetoacetate > acetone
ii. Does not detect B-hydroxybutyrate = #1 ketone formed in diabetic ketoacidosis
iii. False negative RARE since other ketones, athough in small amts, are present
e. False negative: standing urine (room temp), bacterial infection (convert acetoacetate to acetone)
5. Bilirubin: breakdown of Hgb by MPs; conjugated filtered at glomerulus, unconjugated cannot pass
a. Bilirubinuria may be detected before hyperbilirubinemia (low renal threshold)
b. Normal urine negative (except highly concentrated urine in male dogs)
c. Bilirubinuria: (1) intrahepatic or extrahepatic cholestasis, (2) hemolysis
d. Measurement: reagent stripe, tablet (Ictotest) = both use diazoization rxn, tablet more sensitive
e. False negative: standing urine (unstable, breaks down with light)
6. Occult blood: hematuria, hemoglobinuria, or myoglobinuria
a. Measure: reagent strip = peroxidase reaction
i. Intact RBCs appear as green dots on pad; Hgb appears as green color
ii. Can detect as low as 5/ul intact RBCs
iii. If unsure after ammonium sulfate, can do electrophoresis
Discolored urine Centrifuge Sediment Mm damage? Hemolysis? Ammonium sulfate
Hematuria Clears RBCs (but low USG lyse) No No
Hemoglobinuria Does NOT clear No RBCs No Yes Precipitates
Myoglobinuria Does NOT clear No RBCs Yes No Does not precipitate

SEDIMENT EXAM
1. Methodology: method of sample collection affects sediment!!
a. Centrifuge urine at 1500rpm for 5-10mins; do NOT apply brake
b. Supernatant removed, leaving 0.5mL; remix and put 1 drop on slide with coverslip
i. Can air-dry sediment for better recognition of cells and other structures
ii. Can also use standardized plastic well slides
c. Start by adjusting microscope: condenser (close a bit), stage diaphragm, light
d. Examine first with 10x for casts; examine 10 10x fields for average
e. Then examine with 40x for other sediment components. Examine 10 fields
2. Casts: formed in tubules: mucoprotein secreted by tubular cells + cellular elements
a. RARELY in normal urine
b. Form more frequently in acidic urine; will lyse in very dilute urine
c. Indicate tubular damage
Type of cast Normal? Composed of Associated with Appearance
Hyaline casts Low #s in Tubular Proteinuria, passive congestion of
normal-conc microprotein kidneys
urine

Cellular casts NEVER normal Epi cells: Intrarenal dz, acute tubular damage
Tubulointersitial inflammation
WBCs:
Hemorrhage into tubules
RBCs:

Granular casts Rare OK in conc Degeneration of Large #s from renal tubular epi
urine cellular casts degeneration and necrosis
Waxy casts WORST ONE Final stage of Prolonged renal stasis: local tubular
degradation of obstruction, oliguria, CRF
cellular casts

3. Epithelial cells: neoplastic TCC difficult to differentiate from hyperplasia

Epi cells Origin Normal Significance Appearance
Tubular epi Renal tubules Small amt Not reliable, hard to differentiate 30-40um
cells from others
Transitional Renal pelvis, Small amt Large # = inflammation or 30-40um; variable shape
epi cells ureters, bladder, maturation and neoplasia
proximal urethra aging

Normal TCC
Squamous epi Genital tract Voided Indicated contamination from Largest, 50um
cells samples genital tract

4. RBCs: normal <5/hpf; if unsure, add vinegar to lyse RBCs

5. WBCs: normal <5/hpf
a. Large numbers = pyuria; +/- bacteruria
b. Stale urine = degenerate WBCs; dilute urine = LYSE
6. Bacteria: normal urine does not have bacteria until it reaches mid-urethra
a. Bacteria  highly suggestive of UTI; urine culture is gold standard
b. Should be found forming distinct chains/clusters (don’t confuse with Brownian motion)
c. WBCs should be present unless patient is immunocompromised or sample is contaminated
d. >1 million bacteria must be present before they can be seen in urine
7. Sperm: normal in intact males; can be seen in females recently bred
8. Fungi: hyphae or budding yeast
a. Candida may be present in patients with UTI
b. Systemic mycoses found in urine: Blastomyces, Cryptococcus
9. Crystals: some normal; form when saturation of crystalogenic substances; +/- urolithiasis
a. Formation influenced by pH, volume of urine, rate of urine flow
b. Drugs can form some: sulfas, ciprofloxacin
 Appearance Form in . . . Significance Photograph
Struvite = triple Colorless, 3-6 sided Alkaline, Normal
phosphate prisms, coffin lid neutral or Urease producing bacteria
slightly acidic Idiopathic/FLUTD
High P diet in ruminants/pigs
Urolithiasis

Calcium Light yellow to brown, Alkaline to Normal in horses, guinea pigs,

carbonate opaque spheres or neutral rabbits
dumbbell with rare Rarely urolithiasis in ruminants
striations, rice grains,
flowers

Calcium Colorless, picket fence, Acid, neutral, Ethylene glycol toxicity

oxalate occasionally dumbbell or and slightly
monohydrate spindle shaped alkaline urine

Calcium Colorless and clear, Acid, neutral, Can be normal (esp in horses)
oxalate square envelope or and slightly Urolithiasis
dehydrate Maltese cross alkaline urine Less common in EG toxicity

Bilirubin Orange to reddish brown Acid Normal in highly concentrated

needles or granules, urine – in DOGS
sometimes bundle of Hyperbilirubinemia
sticks NEVER normal in cats

Ammonium Yellow brown, opaque, Alkaline, Normal: Dalmatians, English

biurate irregular sphere, thorn neutral or acid bulldogs
apple shape Urolithiasis: liver dz,
portosystemic shunt

Amorphous Can be phosphates, urates Hyperphosphatemia

Others Cholesterol, cystine, sulfa

10. Lipid droplets: round, black, refractile; float under coverslip (different plane)
a. Common in cats; clinical significance unknown
b. Does NOT indicate lipemia
11. Parasite ova: do NOT confuse with fecal contamination

a. Dioctophema renale Capillaria plica

12. Mucous: narrow, twister ribbon-like material
a. Urethral irritation
b. Contamination with genital secretions
c. NORMAL in horse urine
13. Other: algae (protothecosis), plant fiber, pollen grains, Alternaria