Urinalysis

Amanda J. Callens, BS, LVT, Joseph W. Bartges, DVM, PhD*

KEYWORDS
 Urinalysis  Urine sediment examination  Urine chemistries  Urine specific gravity
 Urine dipstick

KEY POINTS
 Urinalysis is a useful laboratory test in documenting urinary tract diseases, and it can also
provide information about other systemic diseases, such as liver failure and hemolysis.
 The collection method and storage time and conditions are the most important preanalyt-
ical sample variables.
 Preanalytical patient variables include physiologic variables or introduced variables
related to treatment or diagnosis.

Urinalysis is an important laboratory test that can be readily performed in veterinary

practice and is considered part of a minimum database. It is useful in documenting
various types of urinary tract diseases and may provide information about other sys-
temic diseases, such as liver failure and hemolysis. Preanalytical sample variables and
patient variables other than those related to disease may influence urinalysis results.
Excellent resources are available for more comprehensive information about urinalysis
in small animals.1–3 Urine may be collected by cystocentesis, urethral catheterization,
or voiding and should be evaluated within 30 minutes. If this is not possible, then it may
be refrigerated for up to 24 hours or submitted to an outside diagnostic laboratory;
however, this may result in crystal precipitation. Refrigeration does not alter urine
pH or specific gravity.

IN-HOUSE VERSUS SEND-OUT TESTING

Most veterinary practices can and should do urinalysis in-house, from the standpoint
of practice economics and quality of care. The test requires only basic laboratory
supplies, including disposable supplies such as a specimen container, disposable
pipettes, conical centrifuge tubes, urine dipsticks, glass slides and coverslips, and
sediment stain, and equipment, including a centrifuge, a refractometer (preferably

The authors have nothing to disclose.

Department of Internal Medicine, Cornell University Veterinary Specialists, 880 Canal Street,
Stamford, CT 06902, USA
* Corresponding author.
E-mail address: [email protected]

Vet Clin Small Anim 45 (2015) 621–637

http://dx.doi.org/10.1016/j.cvsm.2015.02.001 vetsmall.theclinics.com
0195-5616/15/$ – see front matter Ó 2015 Elsevier Inc. All rights reserved.
622 Callens & Bartges

a temperature-compensated veterinary model with a feline-specific scale for specific

gravity), a microscope, and optionally an automated dipstick reader.4–6 It can be
performed easily by clinic technical staff if they are properly trained, making it an
inexpensive, technically feasible test to perform. In-house testing is preferred
because of faster turnaround time and greater accuracy of results, because delayed
analysis is a potential source of introduced error. Another advantage of in-house
testing is that results can be correlated more easily with the rest of the patient’s clin-
ical picture.

Performing the Complete Urinalysis

A description of performing a urinalysis is provided. The sample should be identified
on the sample container using the patient’s clinic number or name and the date and
should be matched to the reporting form. The reporting form should include patient
identification, date, method and time of collection, and any current or recent medica-
tions or diagnostic agents. Transfer 0.5 to 1.0 mL of the sample using sterile tech-
nique to a sterile tube after mixing the sample and submit or store for aerobic
bacteriologic culture. Record the time the urinalysis is performed and whether the
sample was refrigerated on the reporting form. If the sample has been refrigerated,
allow it to reach room temperature or warm it with mixing to either known patient
body temperature or 38 C (101 F). Gently mix the sample by inverting it several times
and transfer 3 to 5 mL to a clear conical centrifuge tube and record the color (yellow,
brown, black, red, white), clarity (clear, cloudy, turbid, flocculent), and odor (normal or
abnormal and describe if possible) on the reporting form. Perform semiquantitative
biochemical analysis by immersing the urine dipstick so that all reagent pads are
covered with urine, start timing the reactions, and drag edge of strip against rim of
tube to remove excess urine. Perform semiquantitative biochemical analysis using
a urine dipstick or automated dipstick reader following manufacturer instructions.
Record results on reporting form. If the urine sample is grossly discolored (eg, gross
hematuria) or turbid, the pigment discolors the reagent pads on the dipstick, making it
difficult to read and giving erroneous interpretation. Attempt to clear the urine by
centrifuging the sample first. If centrifugation results in a red sediment at the bottom
of the conical tube and a clear supernatant, then the dipstick semiquantitative chem-
ical analysis may be performed on the supernatant after transferring it to another tube
using a pipette or by decanting. If the urine sample does not clear with centrifugation,
then interpret results cautiously. Record results for protein, pH, glucose, ketones, and
bilirubin on reporting form; results for specific gravity, urobilinogen, leukocytes, and
nitrite are unreliable, inferior to other tests performed as part of the urinalysis, or of
no clinical significance and should not be reported. Using an appropriate refractom-
eter measure the specific gravity of the urine sample and report on form. If the sample
is discolored or turbid, then the specific gravity can be measured on the supernatant
after centrifugation. Centrifuge 3 to 6 mL of the sample in the conical tube for
5 minutes at 1500 to 2000 rpm (450 g). Do not use the brake to stop the centrifuge
as this may result in suspension of the sediment. Transfer all but 0.5 to 1.0 mL of
the supernatant depending on the volume of urine in the tube to another tube by using
a pipette or by decanting. If the sample cleared with centrifugation, then dipstick
semiquantitative chemical analysis and specific gravity determination may be per-
formed as mentioned previously. Suspend the remaining sediment pellet in the 0.5
to 1.0 mL of supernatant by tapping the conical tip of the tube gently on the table
top. Use less urine to suspend the sediment pellet if less urine was available for
centrifugation. Transfer a drop of the suspension to a clean glass microscope slide
using a pipette and place a glass coverslip over it. There should be enough fluid to
 Urinalysis 623

fill the area under the coverslip, but the coverslip should not float. Examine the slide
immediately using bright field light microscopy optimized for contrast by closing the
condenser diaphragm if the microscope has one or by lowering the condenser. Using
the 10 objective (100 magnification) scan at least 2 edges of the area under the
coverslip and note presence of casts and crystals as amount per low power field
(#/lpf). Using the 40 objective (400 magnification) scan at least 10 fields of the
area under the coverslip for cells (red blood cells [RBCs], white blood cells [WBCs],
epithelial cells), amorphous crystalline material, microorganisms (bacteria, yeast, par-
asites), spermatozoa, and fat droplets and record as amount per high power field
(#/hpf). Estimate numbers if it is difficult to quantitate, and if too numerous to count
record as such. A second slide may be prepared as described, air dried, stained
with a modified Wright stain, Wright-Giemsa stain, or Gram stain, and examined using
the 10 and 40 objectives as described and with immersion oil at 100 (1000
magnification) for identification of bacteria.7–10

STAINED URINE SEDIMENT EXAMINATION

Preanalytical Sample Variables
A urine sample submitted for analysis should be free of any contaminants, collected
into a clean, opaque, airtight, sterile container, and analyzed within 60 minutes of
collection; however, as this is not always the case, the clinician should consider the
influence of preanalytical sample (ie, nonbiologic) variables when interpreting urinaly-
sis results. The most important of these variables are collection method and storage
time and conditions. The collection method is relevant with regard to potential
contamination. For example, samples collected by cystocentesis typically have a
small amount of iatrogenic blood contamination, voided samples often contain con-
taminants from the lower urogenital tract (eg, bacteria, epithelial cells, spermatozoa,
blood) or the environment (eg, bacteria, plant material, debris), samples obtained by
urinary catheterization may contain lower urogenital tract contaminants or iatrogenic
hemorrhage, and samples obtained from the floor or examination table or from a litter
box lined with nonabsorbent material are often contaminated with microbes and
debris. Storage time and conditions may affect urinalysis results. For example, refrig-
eration may promote crystal formation; prolonged storage of samples at room temper-
ature may result in degeneration of cells and casts, altered crystal formation, or
bacterial overgrowth and resultant secondary artifacts (eg, altered pH, decreased
glucose concentration); and exposure to light or air may alter the composition of the
sample.

Preanalytical Patient Variables

Preanalytical patient variables include physiologic variables or introduced variables
related to treatment or diagnosis. Physiologic variables that affect results include
diet, time of day, and reproductive factors. For example, diet may influence urine
pH or crystal formation, samples collected first thing in the morning tend to be more
concentrated and may have altered cellular morphology or microbial viability, and
estrus or recent breeding may influence microscopic findings. Introduced patient vari-
ables include administration of drugs, fluids, or other exogenous agents. For example,
corticosteroids and diuretics interfere with renal concentrating ability, corticosteroids
may also cause proteinuria, antimicrobials may mask urinary tract infections or form
crystals, hydroxyethyl starch interferes with measurement of specific gravity, and
radiographic contrast agents may interfere with measurement of specific gravity or
biochemical analytes or may form crystals.
624 Callens & Bartges

RESULTS OF COMPLETE URINALYSIS

Urine Appearance
Color
Normal urine is typically transparent and yellow or amber on visual inspection. Primar-
ily 2 pigments impart the yellow coloration: urochrome and urobilin. Urochrome is a
sulfur-containing oxidation product of the colorless urochromogen. Urobilin is a
degradation product of hemoglobin. Because the 24-hour urinary excretion of uro-
chrome is constant, highly concentrated urine is amber in color, whereas dilute urine
may be transparent or light yellow color. The intensity of the color is in part related to
the volume of urine collected and the concentration of urine produced; therefore, it
should be interpreted in the context of the urine specific gravity. Caution must be
used not to overinterpret the significance of urine color as part of a complete urinaly-
sis. Significant disease may exist when urine is normal in color. Abnormal urine color
may be caused by the presence of several endogenous or exogenous pigments.
Although the abnormal color indicates a problem, it provides nonspecific information.
Causes of abnormal coloration should be investigated with appropriate laboratory
tests and examination of urine sediment. Detection of abnormal urine color should
prompt questions related to diet, administration of medication, environment, and
collection technique. Knowledge of urine color may also be important in interpreting
colorimetric test results because it may induce interference with the test.
Urine color that is anything other than yellow or amber is abnormal. There are many
potential causes of discolored urine (Fig. 1, Table 1). The most common abnormal
urine color in dogs and cats is red, brown, or black, which may be caused by hema-
turia, hemoglobinuria, myoglobinuria, and bilirubinuria.

Pale yellow urine Urine that is pale yellow or clear in appearance may be normal or
may indicate a polyuric state. Urine may be appropriately dilute if it is associated
with recent consumption or administration of fluids, consumption of a diet containing
low quantities of protein or high quantities of sodium chloride, glucocorticoid excess,
or administration of diuretics. Urine would be considered to be inappropriately

Fig. 1. Urine samples having various colors from clear (3) to pale yellow (1, 5), dark yellow
(4, 6, 7), and red due to hematuria (2).
Table 1
Potential causes of discolored urine

Urine Color Causes Urine Color Causes

Yellow or amber Urochromes Yellow-brown or Bile pigments
Urobilin green-brown
Deep yellow Highly concentrated urine Brown to black (brown or Melanin
Quinacrinea red-brown when viewed Methemoglobin
Nitrofurantoina in bright light in thin layer) Myoglobin
Phenacetina Bile pigments
Riboflavin (large quantities)a Thymola
Phenolsulfonphthalein (acidic urine)a Phenolic compoundsa
Nitrofurantoina
Nitritesa
Naphthalenea
Chlorinated hydrocarbonsa
Aniline dyesa
Homogentisic acida

(continued on next page)

Urinalysis
625
 626
Callens & Bartges
Table 1
(continued )
Urine Color Causes Urine Color Causes
Blue Methylene blue Colorless Very dilute urine (diuretics,
Indigo carmine and indigo blue dyea diabetes mellitus, diabetes
Indicansa insipidus, glucocorticoid excess,
Pseudomonas infectiona fluid therapy, overhydration)
Water-soluble chlorophylla
Rhubarba
Toluidine bluea
Triamterenea
Amitriptylinea
Anthraquinonea
Blue food dyea
Green Methylene blue Milky white Lipid
Dithiazanine Pyuria
Urate crystalluria Crystals
Indigo bluea
Evan bluea
Bilirubin
Biliverdin
Riboflavina
Thymola
Phenola
Triamterenea
Amitriptylinea
Anthraquinonea
Green food dyea
 Red, pink, red-brown, Hematuria Brown Methemoglobin
red-orange, or orange Hemoglobinuria Melanin
Myoglobinuria Sulfasalazinea
Porphyrinuria Nitrofurantoina
Congo red Phenacetina
Phenolsulfonphthalein (following alkalinization) Naphthalenea
Neoprontosil Sulfonamidesa
Warfarin (orange)a Bismutha
Food pigments (rhubarb, beets, blackberries)a Mercurya
Carbon tetrachloridea Feces (rectal-urinary fistula)
Phenazopyridine Fava beansa
Phenothiazinea Rhubarba
Diphenylhydantoina Sorbitola
Bromsulphalein (following alkalinization) Metronidazolea
Chronic heavy metal poisoning (lead, mercury)a Methocarbamola
Rifampina Anthracin catharticsa
Emodina Clofaziminea
Phenindionea Primaquinea
Eosina Chloroquinea
Rifabutina Furazolidonea
Acetazolamidea Copper toxicity
Red food dyea
Orange-yellow Highly concentrated urine
Excess urobilin
Bilirubin
Phenazopyridine
Sulfasalazinea
Fluorescein sodiuma
Flutamidea
Quinacrinea
Phenacetina
2,4-Dichlorophenoxyacetic acida

Urinalysis
Acetazolamidea
Orange food dyea
a
Only observed in human beings.

627
628 Callens & Bartges

concentrated if it were dilute in the presence of dehydration. Diseases that may be

associated with persistently dilute urine include diabetes mellitus, diabetes insipidus,
hyperadrenocorticism, hypoadrenocorticism, hypercalcemia, hyperthyroidism, and
renal failure. If urine is pale yellow or clear, the urine specific gravity is often less
than 1.015. A simple test to determine whether polyuria is persistent is to determine
the urine specific gravity of a sample collected in the morning. Other tests should
include serum biochemical analysis and a complete urinalysis. Additional testing
may include measurement of serum thyroxine concentration, adrenal function testing,
or monitoring urine specific gravity after several days of vasopressin administration.

Red, brown, or black urine Red, brown, or black urine suggests the presence of
blood, hemoglobin, myoglobin, or bilirubin. A positive occult blood reaction is
obtained when urine contains any of these substances. Discoloration of urine may
also result in false-positive reactions on other urine dipstick test pads. Analysis of urine
sediment reveals the presence of RBCs if the discoloration is due to hematuria. If no
RBCs are present on microscopic examination of urine sediment, presence of hemo-
globin, myoglobin, or bilirubin should be suspected. Examination of plasma color may
aid in differentiating these. If the urine is discolored because of myoglobin, the plasma
is clear because myoglobin in plasma is not bound significantly to a carrying protein,
which results in filtration and excretion of myoglobin. If the plasma is pink, it is sugges-
tive of hemoglobin. If the plasma is yellow, it is suggestive of bilirubin; serum bilirubin
concentration should also be increased. Myoglobinuria indicates muscle damage;
serum creatine kinase activity is often increased in this setting. Hemoglobinemia indi-
cates intravascular hemolysis resulting from immune-mediated, parasite-mediated, or
drug-mediated destruction of RBCs. Hyperbilirubinemia may result from liver disease,
post–hepatic obstruction, or hemolysis.
Milky white urine Milky white urine may be due to the presence of WBCs (pyuria),
lipid, or crystals. The more concentrated the urine sample is the more opaque it
may appear. The presence of pyuria secondary to a bacterial urinary tract infection
is the most common cause of milky white urine; however, pyuria may occur because
of inflammation and not be associated with an infection. Lipiduria may be observed in
healthy animals but is frequently observed in cats affected with hepatic lipidosis. Crys-
talluria if heavy and present in a concentrated urine sample may also result in milky
white urine. Microscopic examination of urine sediment aids in differentiation of these
causes.
Clarity
Urine is typically clear but may become less transparent with pigmenturia, crystalluria,
hematuria, pyuria, lipiduria, or when other compounds such as mucus are present.
Depending on the cause, increased turbidity may disappear with centrifugation of
the sample.
Odor
Normal urine has a slight odor of ammonia; however, the odor depends on urine con-
centration. Some species, such as cats and goats, have pungent urine odor because
of urine composition. Bacterial infection may result in a strong odor due to pyuria; a
strong ammonia odor may occur if the bacteria produce urease.

URINE CHEMICAL ANALYSIS

Urine must be at room temperature for accurate measurement of specific gravity and
for chemical analysis. These tests are usually done before centrifugation; however, if
 Urinalysis 629

urine is discolored or turbid, it may be beneficial to perform these tests on the

supernatant.
Specific Gravity
Specific gravity is defined as the ratio of the weight of a volume of liquid to the weight
of an equal volume of distilled water; therefore, it depends on the number, size, and
weight of particles in the liquid. It is different from osmolality, which depends only
on the number of particles in the liquid; measurement of osmolality requires special-
ized instrumentation. Urine specific gravity is determined using a refractometer
designed for veterinary samples, which includes a scale calibrated specifically for
cat urine. Urine specific gravity for species other than cats should be determined using
the scale for dogs. In healthy animals, urine specific gravity is highly variable, depend-
ing on fluid and electrolyte balance of the body. Interpretation of urine specific gravity,
therefore, depends on the clinical presentation and findings of serum chemical anal-
ysis. An animal that is dehydrated or has other causes of prerenal azotemia has hyper-
sthenuric urine with a specific gravity of 1.025 to 1.040 (depending on the species).
Dilute urine in a dehydrated or azotemic animal is abnormal and could be caused
by renal failure, hypoadrenocorticism or hyperadrenocorticism, hypercalcemia, dia-
betes mellitus, hyperthyroidism, diuretic therapy, or diabetes insipidus. Glucosuria
increases the urine specific gravity despite increased urine volume.
Semiquantitative, Colorimetric Reagent Strips
Reagent strips such as Multistix (Siemens Healthcare, Erlangen, Germany) or Chem-
strip (Roche Diagnostics, Indianapolis, IN) can be used to perform several semiquan-
titative chemical evaluations simultaneously. They are used routinely to determine
urine pH, protein, glucose, ketones, bilirubin/urobilinogen, and occult blood. Some re-
agent strips include test pads for leukocyte esterase (for detection of WBCs), nitrite
(for detection of bacteria), and (urine specific gravity) USG; these are not valid in ani-
mals and should not be used. Reagent strips are adversely affected by moisture and
have a limited shelf life. Bottles should be kept tightly capped, and unused strips
should be discarded after their expiration date.
pH
Urine pH is typically acidic in dogs and cats and alkaline in horses and ruminants but
varies depending on diet, medications, or presence of disease. Reagent strip colori-
metric test pads for pH determination are accurate to within 0.5 pH units.11 For
example, a reading of 6.5 means the actual pH is likely to be between 6.0 and 7.0.
Portable pH meters are more accurate than pH colorimetric test pads. A bacterial uri-
nary tract infection with a urease-producing microbe results in alkaluria. Urine pH
affects crystalluria because some crystals, such as struvite, form in alkaline urine,
whereas other crystals, such as cystine, form in acidic urine.
Protein
The protein test pad uses a color indicator (tetrabromophenol blue), which detects pri-
marily albumin in urine. Results range from 10 to 1000 mg/dL. Proteinuria can occur
from prerenal (fever, strenuous exercise, seizures, extreme environmental tempera-
ture, and hyperproteinemia), renal (primarily glomerular and occasionally tubular dis-
ease), or postrenal (inflammation, hemorrhage, and infection) causes. A positive
reaction must be interpreted in light of USG, pH, and urine sediment examination.
For example, a trace amount of protein in concentrated urine is less significant than
a trace amount of protein in dilute urine. Alkaluria gives a false-positive reaction. Like-
wise, presence of other proteins, such as Bence-Jones proteins, gives false-negative
630 Callens & Bartges

results. Proteinuria can be measured using sulfosalicylic acid precipitation, which de-
tects albumin and globulins; however, it is not accurate in dogs and cats. If proteinuria
is present with an inactive urine sediment, its significance can be verified and quanti-
tated by dividing the urine protein concentration by the urine creatinine concentration
(urine protein to urine creatinine ratio; UP:UC). Interpretation of UP:UC is as follows:
less than 0.5:1.0 (dogs) and less than 0.4:1.0 (cats) is normal, 0.4 or 0.5 to 1.0:1.0 is
questionable, and greater than 1.0:1.0 is abnormal. With primary renal azotemia,
UP:UC greater than 0.4:1.0 in cats and 0.5:1.0 in dogs is considered abnormal.12 A
semiquantitative microalbuminuria test is available to detect urinary albumin in the
range of 1 to 30 mg/dL. It uses enzyme-linked immunosorbent assay technology spe-
cific for canine or feline albumin. Because of minor species differences to albumin,
there are different kits for dogs and cats. The microalbuminuria test detects lower con-
centrations of albumin than a standard dipstick test pad. Hematuria must be macro-
scopic to increase the microalbuminuria or UP:UC; however, pyuria increases both.

Glucose
Glucose is detected by a glucose oxidase enzymatic reaction that is specific for
glucose. Glucosuria is not present normally because the renal threshold for glucose
is greater than 180 mg/dL in most species and greater than 240 mg/dL in cats. With
euglycemia, the amount of filtered glucose is less than the renal threshold and all
the filtered glucose is reabsorbed in the proximal renal tubules. Glucosuria can result
from hyperglycemia (due to diabetes mellitus, excessive endogenous or exogenous
glucocorticoids, or stress) or from a proximal renal tubular defect (such as primary
renal glucosuria or Fanconi syndrome). If glucosuria is present, blood glucose concen-
tration should be determined. False-negative results can occur with high urinary con-
centrations of ascorbic acid (vitamin C) or with formaldehyde (a metabolite of the
urinary antiseptic, methenamine, which may be used for prevention of bacterial urinary
tract infections). False-positive results may occur if the sample is contaminated with
hydrogen peroxide, chlorine, or hypochlorite (bleach).

Ketones
Ketones are produced from fatty acid metabolism and include acetoacetic acid,
acetone, and b-hydroxybutyrate. The ketone test pad detects acetone and aceto-
acetic acid but not b-hydroxybutyrate. The test pad contains nitroprusside that
reacts with acetoacetic acid and acetone to cause a purple color change; it is
more sensitive to acetoacetic acid than acetone. Ketonuria is associated with pri-
mary ketosis (ruminants), ketosis secondary to diabetes mellitus (small animals),
consumption of low-carbohydrate diets (especially in cats), and occasionally pro-
longed fasting or starvation. A false-positive reaction can occur with the presence
of reducing substances in the urine.

Bilirubin/Urobilinogen
When hemoglobin is degraded, the heme portion is converted to bilirubin, which is con-
jugated in the liver and excreted in bile. Some conjugated bilirubin is filtered by the
glomerulus and excreted in urine. The kidney can metabolize hemoglobin to bilirubin
and secrete it in dogs but not in cats. Male dogs have a higher secretory ability than fe-
male dogs. Dipstick reagent pads use diazonium salts to create a color change and are
more sensitive to conjugated bilirubin than unconjugated bilirubin. Bilirubinuria occurs
when the level of conjugated bilirubin exceeds the renal threshold as with liver disease
or hemolysis. In dogs with concentrated urine, a small amount of bilirubin can be
normal. Pigmenturia and phenothiazine may result in a false-positive reaction; false-
negative reactions may occur with large amounts of urinary ascorbic acid (vitamin C).
 Urinalysis 631

Urobilinogen, formed from bilirubin by intestinal microflora, is absorbed into the por-
tal circulation and is excreted renally. A small amount of urinary urobilinogen is normal.
Increased urinary urobilinogen level occurs with hyperbilirubinemia; a negative test
result may be observed with biliary obstruction; however, the test is not specific
enough to be clinically useful.
Occult blood
The occult blood test pad uses a pseudoperoxidase method to detect intact RBCs,
hemoglobin, and myoglobin. A positive reaction can be due to hemorrhage (hematu-
ria), intravascular hemolysis (hemoglobinuria), or myoglobinuria. The last 2 processes
can be distinguished by examination of plasma; plasma appears pink to red after intra-
vascular hemolysis, whereas myoglobin is rapidly cleared from plasma, resulting in
clear plasma. As with other colorimetric test pads, discolored urine may yield false-
positive results. A positive result should be interpreted with microscopic examination
of urine sediment.

URINE SEDIMENT

Microscopic examination of urine sediment should be part of a routine urinalysis. For

centrifugation, 3 to 5 mL of urine is transferred to a conical centrifuge tube. Urine is
centrifuged at 1000 to 1500 rpm for 5 minutes. The supernatant is decanted, leaving
0.5 to1.0 mL of urine and sediment in the tip of the conical tube. The sediment is resus-
pended by tapping the tip of the conical tube against the table several times. A few
drops of the sediment are transferred to a glass slide, and a coverslip is applied.
Examination of unstained urine is recommended for routine samples. Microscopic
examination is performed at 10 objective (for crystals, casts, and cells) and 40
objective (for cells and bacteria) magnifications. Contrast of the sample is enhanced
by closing the iris diaphragm and lowering the condenser of the microscope. Stains
such as Sedi-Stain and new methylene blue can be used to aid in cell identification
but may dilute the specimen and introduce artifacts such as stain precipitate and crys-
tals. Use of a modified Wright stain increases the sensitivity, specificity, and positive
and negative predictive values for detection of bacteria.
Red Blood Cells
In an unstained preparation, RBCs are small and round and have a slight orange tint
and a smooth appearance (Fig. 2). Normal urine should contain less than 5 RBCs
per field at 400 magnification (40 objective). Increased RBC count in urine (hema-
turia) indicates hemorrhage somewhere in the urogenital system; however, sample
collection by cystocentesis or catheterization may induce hemorrhage.
White Blood Cells
WBCs are slightly larger than RBCs and have a grainy cytoplasm (see Fig. 2; Fig. 3).
Normal urine should contain less than 5 WBCs per field at 400 magnification (40
objective). Increased WBC count (pyuria) can occur because of inflammation, infec-
tion, trauma, or neoplasia. Catheterization or collection of voided urine may introduce
a few WBCs from the urogenital tract.
Epithelial Cells
Transitional epithelial cells, a common urine contaminant derived from the bladder and
proximal urethra, resemble WBCs but are larger. They have a greater amount of grainy
cytoplasm and a round, centrally located nucleus. In a voided urine sample, squamous
epithelial cells may be observed. They are large, oval to cuboidal, and may or may not
632 Callens & Bartges

Fig. 2. Red blood cells and 1 white blood cell (arrow).

contain a nucleus. Occasionally, neoplastic transitional cells may be observed in an

animal with a transitional cell carcinoma. Neoplastic squamous cells may be observed
in an animal with a squamous cell carcinoma.

Cylindruria (Casts)
Casts are elongated, cylindrical structures formed by mucoprotein congealing within
renal tubules and may contain cells (Fig. 4). Hyaline casts are pure protein precipi-
tates, are transparent, have parallel sides and rounded ends, and are composed of
mucoprotein. They may occur with fever, exercise, and renal disease. Epithelial
cellular casts form from entrapment of sloughed tubular epithelial cells in the

Fig. 3. White blood cells (neutrophils) and bacteria.

 Urinalysis 633

Fig. 4. Granular cast.

mucoprotein; they may be observed with renal tubular disease. Granular casts are
thought to represent degenerated epithelial cellular casts. Waxy casts have a granular
appearance and are thought to arise from degeneration of long-standing granular
casts. They typically have sharp borders with broken ends. Other cellular casts include
erythrocyte casts and WBC casts and are always abnormal. Erythrocyte casts form
because of renal hemorrhage. WBC casts occur because of renal inflammation, as
with pyelonephritis. Fatty casts are not common but can be observed with disorders
of lipid metabolism, such as diabetes mellitus. A few hyaline or granular casts are
considered normal. However, the presence of cellular casts or other casts in high
numbers indicates renal damage and may be one of the earliest abnormalities deter-
mined by laboratory tests noted with toxic damage to renal epithelial cells (eg, genta-
micin, amphotericin B).

Infectious Organisms
The presence of bacteria in urine collected by cystocentesis indicates infection. Small
numbers of bacteria from the lower urogenital tract may contaminate voided samples
or samples collected by catheterization and do not indicate infection. Bacterial rods
are most easily identified in urine sediment (see Fig. 3). Particles of debris may be
mistaken for bacteria. Suspected bacteria can be confirmed by staining urine sedi-
ment with Gram stain or modified Wright stain; however, aerobic culture is best to
confirm a bacterial urinary tract infection. Rarely, yeast and fungi (Fig. 5) and parasitic

Fig. 5. Fungal organism (Blastomyces spp) and white blood cells (neutrophils).
634 Callens & Bartges

ova may be observed in urine sediment. Their presence is not always associated with
clinical disease. Parasitic ova observed include Stephanus dentatus, Capillaria plica,
Capillaria felis (Fig. 6), and Dioctophyma renale. In addition, microfilariae of Dirofilaria
immitis may be observed in urine sediment.
Crystals
Many urine sediments contain crystals. The type of crystal present depends on urine
pH, concentration of crystallogenic materials, urine temperature, and length of time
between urine collection and examination. Crystalluria is not synonymous with uro-
lithiasis and is not necessarily pathologic. Furthermore, uroliths may form without
observed crystalluria. Struvite crystals (Fig. 7) are commonly observed in canine
and feline urine. Struvite crystalluria in dogs is not a problem unless there is a concur-
rent bacterial urinary tract infection with a urease-producing microbe. Without an
infection, struvite crystals in dogs are not associated with struvite urolith formation.
However, struvite uroliths form in some animals (eg, cats) without a bacterial urinary
tract infection. In these animals, struvite crystalluria may be pathologic. Struvite crys-
tals appear typically as coffin lids or prisms; however, they may be amorphous. Cal-
cium oxalate crystalluria occurs less commonly in dogs and cats; if persistent, it may
indicate an increased risk for calcium oxalate urolith formation. However, calcium
oxalate and calcium carbonate crystalluria is common in healthy horses and cattle.
Calcium oxalate dihydrate crystals appear as squares with an X in the middle
(Fig. 8) or as envelope shaped. Calcium oxalate monohydrate crystals are dumb-
bell shaped. An unusual form of calcium oxalate crystals is typically seen in associ-
ation with ethylene glycol toxicity. These crystals occur in neutral to acidic urine.
They are small, flat, and colorless and are shaped like picket fence posts. Ammonium
acid urate crystals suggest liver disease (eg, portosystemic shunt). These crystals
occur in acidic urine and are yellow-brown spheres with irregular, spiny projections
(Fig. 9); however, they may also be amorphous. Certain species, such as birds and
reptiles, and certain breeds of dogs, specifically Dalmatians, can normally have
ammonium acid urate crystalluria. Cystine crystals are 6 sided and of variable size

Fig. 6. Capillaria felis ovum.

 Urinalysis 635

Fig. 7. Struvite crystals, red blood cells, and spermatozoon (arrow).

Fig. 8. Calcium oxalate dihydrate crystals.

Fig. 9. Ammonium urate crystals.

636 Callens & Bartges

Fig. 10. Cystine crystals.

(Fig. 10). They occur in acidic urine. Presence of cystine crystals represents a prox-
imal tubular defect in amino acid reabsorption. Cystinuria has been reported to occur
in many breeds of dogs and rarely in cats. Dachshunds, Newfoundlands, English bull-
dogs, and Scottish terriers have a high incidence of cystine urolithiasis. Bilirubin crys-
tals occur with bilirubinuria; however, they may be normal in small numbers in dogs.

Lipids
Fat droplets are commonly present in urine from dogs and cats and may be mistaken
for RBC. They often vary in size and tend to float on a different plane of focus than the
remainder of the sediment. They are not considered to be pathologic.

Spermatozoa
Spermatozoa may be observed normally in urine collected from male dogs (see Fig. 7).

Plant Material
Occasionally, plant material may be observed in urine samples collected by voiding
(Fig. 11). When present, it indicates contamination of the urine sample and is not
pathologic.

Fig. 11. Plant pollen in a voided urine sample.

 Urinalysis 637

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