Physiology Lab Report 10 - Urinalysis
Physiology Lab Report 10 - Urinalysis
Physiology Lab Report 10 - Urinalysis
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OBJECTIVE
To conduct the tests those are conducted to examine the physical, chemical and
microscopic properties of the urine.
INTRODUCTIONS
When cells in the body break down proteins into forms they can utilize, they
produce ammonia wastes that the liver turns into urea (a chemical compound of carbon,
hydrogen, nitrogen, and oxygen). When cells break down carbohydrates, they produce
water and carbon dioxide as waste products. If these useless waste products were allowed
to accumulate in the body, they would become dangerous to the body's health. The body
eliminates these wastes in a process known as excretion. The body system most
responsible for waste excretion is the urinary system, which eliminates water, urea, and
other waste products from the body in the form of urine.
The main organs of the urinary system are the kidneys, which form urine. The
other parts of the system include the ureter, the urinary bladder, and the urethra, neither
form urine nor change its composition. They are merely structures that transport urine
from the kidneys to the outside of the body. Urine is formed in the kidneys as a result of
three processes. There are filtration, reabsorption, and secretion. Filtration takes place in
the renal corpuscles; reabsorption and secretion take place in the renal tubules.
The most cost-effective device used to screen urine is a paper or plastic dipstick.
This microchemistry system has been available for many years and allows qualitative and
semi-quantitative analysis within one minute by simple but careful observation. The color
change occurring on each segment of the strip is compared to a color chart to obtain
results. However, a careless doctor, nurse, or assistant is entirely capable of misreading or
misinterpreting the results. Microscopic urinalysis requires only a relatively inexpensive
light microscope.
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Generally, urinalysis consists of physical tests, chemical tests, and microscopic
analysis as follows:
A. PHYSICAL TEST
A red or red-brown (abnormal) color could be from a food dye, eating fresh beets,
a drug, or the presence of either hemoglobin or myoglobin. If the sample contained many
red blood cells, it would be cloudy as well as red.
B. CHEMICAL TEST
These urinalysis test strips, URS-K (Ketones) URS-3 (Glucose, Protein, pH) and
URS-10 (Glucose, Protein pH, Leukocytes, Nitrites, Ketones, Bilirubin, Blood,
Urobilinogen, and Specific Gravity) are simple, easy to use reagent strips for the
detection of key diagnostic chemical markers in human urine. They are the same test
strips used routinely by doctors, laboratories and healthcare professionals in preliminary
diagnosis of, and initial screening for potential health problems. URS-Strips are plastic
strips to which chemically specific reagent pads are affixed.
The reagent pads react with the sample urine to provide a standardized visible
color reaction within 30 seconds to one minute depending on the specific panel screen.
The color is then visually compared to the included color chart to determine the level of
each chemical factor. Test results may provide useful information regarding carbohydrate
(sugar) metabolism (diabetes), kidney function, acid-base balance, bacteriuria, occult
blood, high leukocytes (infection) and other conditions of overall health. The strips are
ready to use upon removal from the vial and the entire reagent strip is disposable. URS-
K, 3 & 10 reagent test strips are packed 100 tests to a vial with approximately one (1)
year shelf life. Each vial comes with a complete abstract on the chemical and biological
properties of the test including a color chart for rapid visual diagnosis.
Urinalysis test strips can be used to determine the following aspects of urine:
Specific gravity
The specific gravity of urine can be affected by a range of diseases and disorders.
Low specific gravity (below 1.005) is associated with diabetes insipidus, nephrogenic
diabetes insipidus, acute tubular necrosis, and inflammation of the upper urinary tract
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(pyelonephritis). In fixed specific gravity, the specific gravity of the urine remains at
1.010 no matter how much fluid the person drinks. This condition occurs in patients who
have chronic inflammation of the small blood vessels in the kidneys (glomerulonephritis)
and serious kidney damage. High specific gravity (above 1.035) occurs in patients who
are in shock or who suffer from nephrotic syndrome, dehydration, acute
glomerulonephritis, congestive heart failure, or liver failure.
pH
A pH factor greater than 7 (more alkaline) may result from Fanconi's syndrome,
urinary tract infections, or metabolic or respiratory alkalosis. A pH factor below 7 (more
acid) may be due to fever, PKU, the secretion of homogentisic acid in the urine
(alkaptonuria), and acidosis.
Protein
Glucose
A high level of glucose and other sugars in the urine (glycosuria) is often a
symptom of diabetes mellitus. Glycosuria can also be caused by advanced kidney disease,
Cushing's syndrome, impaired tubular reabsorption, shock, a rare tumor of the adrenal
gland (pheochromocytoma), or cancer of the pancreas. Milk in the urine is normal if a
woman is pregnant, has just given birth, or is breastfeeding. On the other hand, rare
hereditary metabolic disorders are indicated when urine contains fruit sugar (fructose),
milk sugar (galactose), or a simple sugar called pentose.
Ketones
Bilirubin
Urobilinogen
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Low levels of urobilinogen in the urine may be a result of antimicrobial therapy,
inflammatory diseases, kidney disease, severe diarrhea, or blocked bile ducts.
Nitrite
A positive nitrite test indicates that bacteria may be present in significant numbers
in urine. Gram negative rods such as E. coli are more likely to give a positive test.
Red blood cells in the urine can be due to vigorous exercise or exposure to toxic
chemicals. Bloody urine can also be a sign of bleeding in the genitourinary tract as a
result of systemic bleeding disorders, various kidney diseases, bacterial infections,
parasitic infections including malaria, obstructions in the urinary tract, scurvy, subacute
bacterial endocarditis, traumatic injuries, and tumors.
A high number of white blood cells in the urine is usually a symptom of urinary
tract infection. A large number of cells from tissue lining (epithelial cells) can indicate
damage to the small tubes that carry material into and out of the kidneys.
C. MICROSCOPIC ANALYSIS
The sediment obtained after centrifugation is first examined under low power to
identify most crystals, casts, squamous cells, and other large objects. The numbers of
casts seen are usually reported as number of each type found per low power field (LPF).
Example: 5-10 hyaline casts/L casts/LPF. Since the number of elements found in each
field may vary considerably from one field to another, several fields are averaged. Next,
examination is carried out at high power to identify crystals, cells, and bacteria. The
various types of cells are usually described as the number of each type found per average
high power field (HPF). Example: 1-5 WBC/HPF.
The main sediments that we can observe in the microscopic analysis of urine
sample are cells, casts, crystals and also other organisms or artifacts. The presence of
bacteria, parasites, or yeast cells in the urine may be a symptom of urinary tract infection
or contamination of the external genitalia.
MATERIALS
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Slides and Cover Slips
Pasteur Pipette
PROCEDURES
A. Physical Test
B. Chemical Test
C. Microscopic analysis
1. The urine sample is mixed and shaked well so that the components in the
urine are in equilibrium.
2. Approximately 7ml of both normal and abnormal samples are being poured
into the test tubes and centrifuged for 5 minutes under a speed of 1500rpm by
using the centrifuge.
3. About 6 ml of the supernatant is discarded. The remaining 1ml would contain
all of the sediments required.
4. Upon removal, the 1ml of remaining liquid is mixed.
5. 1 drop of the specimen is put onto the slide and dried. Then one drop of
Methylene Blue reagent is being dripped onto the slide and dried.
6. The urine sample is viewed under the microscope with a low power of x 10
and a high power of x 20.
7. The solid substances are identified.
8. The types of substances are drawn.
9. The experiment is repeated by using the sample from an ill patient.
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RESULTS
DISCUSSION
1. Bacteria proliferate
2. Cells autolyse and cytological features deteriorate rapidly
3. Dissolve of casts if the urine is alkaline
4. Increase in pH because of bacteria action breakdown of urea
5. Sunlight from affecting bilirubin and urobilinogen
6. Overgrowth of contaminating microorganisms
7. Loss of ketone bodies
8. Crystallization of solutes and decrease the clarity
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A. PHYSICAL TESTS
Colour
Urine color will vary between species, but it is normally some shade of yellow
depending on the concentration. Colour is due to urochrome (pigment produced from
breakdown of bile) and urobilin (from breakdown of hemoglobin). The colour shows how
concentrated is the urine. Abnormal color changes in the urine could be due to drugs,
increased urinary pigments or red blood cells. Red to reddish-brown could be due to
either hematuria , hemoglobinuria, or myoglobinuria. Yellow-green to yellow-brown is
associated with bilirubinuria. Occasionally, unusual colors may be caused by dyes
associated with food or drugs.
In this experiment, both samples are pale yellow and yellowish in color which
indicates they are normal.
Clarity
Urine may be cloudy (turbid) because it contains red or white blood cells, casts, or
crystals, bacteria, fat, mucus, digestive fluid (chyle), or pus from a bladder or kidney.
Crystallization or precipitation of salts upon standing at room temperature or in the
refrigerator may also be a cause of turbidity. Clearing of the specimen after addition of a
small amount of acid indicates that precipitation of salts is the probable cause of tubidity.
In this experiment, the both sampes are clear so they are considered normal.
B. CHEMICAL TESTS
pH
The glomerular filtrate of blood plasma is usually acidified by renal tubules and
collecting ducts from a pH of 7.4 to about 6 in the final urine. However, depending on the
acid-base status, urinary pH may range from as low as 4.5 to as high as 8.0. The change
to the acid side of 7.4 is accomplished in the distal convoluted tubule and the collecting
duct.
Specific Gravity
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measure specific gravity in approximations. Most laboratories measure specific gravity
with a refractometer. Specific gravity between 1.002 and 1.035 on a random sample
should be considered normal if kidney function is normal. Since the specific gravity of
the glomerular filtrate in Bowman's space ranges from 1.007 to 1.010, any measurement
below this range indicates hydration and any measurement above it indicates relative
dehydration.
If specific gravity is not > 1.022 after a 12 hour period without food or water,
renal concentrating ability is impaired and the patient either has generalized renal
impairment or nephrogenic diabetes insipidus. In end-stage renal disease, sp gr tends to
become 1.007 to 1.010.Any urine having a specific gravity over 1.035 is either
contaminated, contains very high levels of glucose, or the patient may have recently
received high density radiopaque dyes intravenously for radiographic studies or low
molecular weight dextran solutions. Subtract 0.004 for every 1% glucose to determine
non-glucose solute concentration.
Protein
Dipstick screening for protein is done on whole urine, but semi-quantitative tests
for urine protein should be performed on the supernatant of centrifuged urine since the
cells suspended in normal urine can produce a falsely high estimation of protein.
Normally, only small plasma proteins filtered at the glomerulus are reabsorbed by the
renal tubule. However, a small amount of filtered plasma proteins and protein secreted by
the nephron (Tamm-Horsfall protein) can be found in normal urine. Normal total protein
excretion does not usually exceed 150 mg/24 hours or 10 mg/100 ml in any single
specimen. More than 150 mg/day is defined as proteinuria. Proteinuria > 3.5 gm/24 hours
is severe and known as nephrotic syndrome.
In rough terms, trace positive results (which represent a slightly hazy appearance
in urine) are equivalent to 10 mg/100 ml or about 150 mg/24 hours (the upper limit of
normal). 1+ corresponds to about 200-500 mg/24 hours, a 2+ to 0.5-1.5 gm/24 hours, a
3+ to 2-5 gm/24 hours, and a 4+ represents 7 gm/24 hours or greater.
In this experiment, the protein is found in the sample 2 but not in sample 1. This
indicates that the subject number 2 may have proteinuria or the presence of an excess of
serum proteins in the urine. Proteinuria may be a sign of renal (kidney) damage. Since
serum proteins are readily reabsorbed from urine, the presence of excess protein indicates
either an insufficiency of absorption or impaired filtration. Diabetics usually suffer from
damaged nephrons and develop proteinuria. With severe proteinuria, general
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hypoproteinemia can develop which results in diminished oncotic pressure. Symptoms of
diminished oncotic pressure may include ascites, edema, and hydrothorax.
Glucose
Less than 0.1% of glucose normally filtered by the glomerulus appears in urine (<
130 mg/24 hr). Glycosuria (excess sugar in urine) generally means diabetes mellitus.
Dipsticks employing the glucose oxidase reaction for screening are specific for glucos
glucose but can miss other reducing sugars such as galactose and fructose. For this
reason, most newborn and infant urines are routinely screened for reducing sugars by
methods other than glucose oxidase (such as the Clinitest, a modified Benedict's copper
reduction test).
In this experiment, glucose is present in sample 2 but not sample 1. This means
that person might be a diabetes patient because high level of glucose and other sugars in
the urine (glycosuria) is often a symptom of diabetes mellitus. Glycosuria can also be
caused by advanced kidney disease, Cushing's syndrome, impaired tubular reabsorption,
shock, a rare tumor of the adrenal gland (pheochromocytoma), or cancer of the pancreas.
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Ketones
Nitrite
A positive nitrite test indicates that bacteria may be present in significant numbers
in urine. Gram negative rods such as E. coli are more likely to give a positive test.
Red blood cells can enter the urine from the vagina in menstruation or from the
trauma of bladder catherization. Red blood cells in the urine otherwise may be due to
many causes including kidney damage, tumors eroding the urinary tract, stones, and
urinary tract infections.
In this experiment, although white blood cells are detected in sample 1, it is still
be considered normal as white blood cells from the vagina or the opening of the urethra
(in males, too) can contaminate a urine sample. Such contamination aside, the presence of
abnormal numbers of white blood cells in the urine is important. It can mean there is
kidney disease or an infection of the kidney, bladder, or urinary tubes (upper or lower
urinary tract). The presence of abnormal numbers of white cells in the urine is referred to
as pyuria (pus in the urine).
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When stimulated by eating, bile (including the conjugated bilirubin) is excreted
into the small intestine. In the later portions of the small intestine (ileum) and the colon,
about half of the bilirubinglucuronide is converted into urobilinogen. Urobilinogen is
either reabsorbed or converted by the presence of oxygen to stercobilin. The stercobilin
and remaining bilirubinglucuronide are excreted in the feces. These two metabolites of
bilirubin are what give feces their characteristic brown color. Small amounts of
urobilinogen remaining in the blood are filtered by the kidneys, ending up in the urine as
urobilin. This bilirubin metabolite gives urine its characteristic yellow color. In diseases
where too much haemoglobin is broken down or the removal of bilirubin does not
function properly, the accumulating bilirubin in the body causes jaundice.
In this experiment, both samples are normal for bilirubin and urobilinogen tests.
C. MICROSCOPIC ANALYSIS
Type of urine sediment is observed in this experiment. Here are various types of
sediments that might present in the sample:
Hematuria is the presence of abnormal numbers of red cells in urine due to:
glomerular damage, tumors which erode the urinary tract anywhere along its length,
kidney trauma, urinary tract stones, renal infarcts, acute tubular necrosis, upper and lower
uri urinary tract infections, nephrotoxins, and physical stress. Red cells may also
contaminate the urine from the vagina in menstruating women or from trauma produced
by bladder catherization. Theoretically, no red cells should be found, but some find their
way into the urine even in very healthy individuals. However, if one or more red cells can
be found in every high power field, and if contamination can be ruled out, the specimen is
probably abnormal.
RBC's may appear normally shaped, swollen by dilute urine (in fact, only cell
ghosts and free hemoglobin may remain), or crenated by concentrated urine. Both
swollen, partly hemolyzed RBC's and crenated RBC's are sometimes difficult to
distinguish from WBC's in the urine. In addition, red cell ghosts may simulate yeast. The
presence of dysmorphic RBC's in urine suggests a glomerular disease such as a
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glomerulonephritis. Dysmorphic RBC's have odd shapes as a consequence of being
distorted via passage through the abnormal glomerular structure.
Pyuria refers to the presence of abnormal numbers of leukocytes that may appear
with infection in either the upper or lower urinary tract or with acute glomerulonephritis.
Usually, the WBC's are granulocytes. White cells from the vagina, especially in the
presence of vaginal and cervical infections, or the external urethral meatus in men and
women may contaminate the urine.
If two or more leukocytes per each high power field appear in non-contaminated
urine, the specimen is probably abnormal. Leukocytes have lobed nuclei and granular
cytoplasm.
Epithelial Cells
Renal tubular epithelial cells, usually larger than granulocytes, contain a large
round or oval nucleus and normally slough into the urine in small numbers. However,
with nephrotic syndrome and in conditions leading to tubular degeneration, the number
sloughed is increased.
When lipiduria occurs, these cells contain endogenous fats. When filled with
numerous fat droplets, such cells are called oval fat bodies. Oval fat bodies exhibit a
"Maltese cross" configuration by polarized light microscopy.
Transitional epithelial cells from the renal pelvis, ureter, or bladder have more
regular cell borders, larger nuclei, and smaller overall size than squamous epithelium.
Renal tubular epithelial cells are smaller and rounder than transitional epithelium, and
their nucleus occupies more of the total cell volume.
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Squamous epithelial cells from the skin surface or from the outer urethra can
appear in urine.
Casts
Urinary casts are formed only in the distal convoluted tubule (DCT) or the
collecting duct (distal nephron). The proximal convoluted tubule (PCT) and loop of
Henle are not locations for cast formation. Hyaline casts are composed primarily of a
mucoprotein (Tamm-Horsfall protein) secreted by tubule cells. The Tamm-Horsfall
protein secretion (green dots) is illustrated in the diagram below, forming a hyaline cast
in the collecting duct:
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incorporated. An example of glomerular inflammation with leakage of RBC's to produce
a red blood cell cast is shown in the diagram below:
The factors which favor protein cast formation are low flow rate, high salt
concentration, and low pH, all of which favor protein denaturation and precipitation,
particularly that of the Tamm-Horsfall protein. Protein casts with long, thin tails formed
at the junction of Henle's loop and the distal convoluted tubule are called cylindroids.
Hyaline casts can be seen even in healthy patients.
Red blood cells may stick together and form red blood cell casts. Such casts are
indicative of glomerulonephritis, with leakage of RBC's from glomeruli, or severe tubular
damage.
White blood cell casts are most typical for acute pyelonephritis, but they may also
be present with glomerulonephritis. Their presence indicates inflammation of the kidney,
because such casts will not form except in the kidney.
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When cellular casts remain in the nephron for some time before they are flushed
into the bladder urine, the cells may degenerate to become a coarsely granular cast, later a
finely granular cast, and ultimately, a waxy cast. Granular and waxy casts are be believed
to derive from renal tubular cell casts. Broad casts are believed to emanate from damaged
and dilated tubules and are therefore seen in end-stage chronic renal disease.
The so-called telescoped urinary sediment is one in which red cells, white cells,
oval fat bodies, and all types of casts are found in more or less equal profusion. The
conditions which may lead to a telescoped sediment are: 1) lupus nephritis 2) malignant
hypertension 3) diabetic glomerulosclerosis, and 4) rapidly progressive
glomerulonephritis.
In end-stage kidney disease of any cause, the urinary sediment often becomes very scant
because few remaining nephrons produce dilute urine.
Bacteria
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Multiple organisms reflect contamination. However, the presence of any organism in
catheterized or suprapubic tap specimens should be considered significant.
Yeast
Yeast cells may be contaminants or represent a true yeast infection. They are
often difficult to distinguish from red cells and amorphous crystals but are distinguished
by their tendency to bud. Most often they are Candida, which may colonize bladder,
urethra, or vagina.
Crystals
Common crystals seen even in healthy patients include calcium oxalate, triple
phosphate crystals and amorphous phosphates.
Uric acid
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Uric acid is the final oxidation product of purine metabolism in the human body
and is found in small amounts in urine. Purine precursors include xanthine and
hypoxanthine, which are converted to uric acid by xanthine oxidase. Uric acid is the end
product of nitrogen metabolism in birds and reptiles, whereas in humans and many other
higher animals the main product of nitrogen detoxification is urea, and in fish, bacteria
and protozoa, it is ammonia.
In animals that produce uric acid in high quantities, it is excreted in feces as a dry
mass. While this compound is produced through a complex and energetically costly
metabolic pathway (in comparison to other nitrogenated wastes), its elimination
minimizes water loss. It is therefore commonly found in the excretions of animals that
live in very dry environments.
Humans produce only small quantities of uric acid with excess accumulation
leading to a type of arthritis known as gout. In human blood, uric acid concentrations
between 3.6 and 8.3 mg/dL are considered normal by the American Medical Association,
although significantly lower levels are common in vegetarians due to a decreased intake
of purine-rich meat. The disease gout in humans is associated with abnormal levels of
uric acid in the system. Saturation of uric acid in the human blood stream may result in
one form of kidney stones when the acid crystallizes into solid inside the kidney. A
percentage of gout patients eventually get uric kidney stones.
Others
General "crud" or unidentifiable objects may find their way into a specimen,
particularly those that patients bring from home. Spermatozoa can sometimes be seen.
Rarely, pinworm ova may contaminate the urine. In Egypt, ova from bladder infestations
with schistosomiasis may be seen.
From this experiment, it is observed that several sediments are present in each of
the samples. The sediments have been shown in the results. All the sediments that present
are normal for both samples.
CONCLUSION
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aware that they may have a problem. It is used to detect urinary tract infections (UTI) and
other disorders of the urinary tract.
REFERENCES
1. http://www.medicinenet.com/script/main/art.asp?articlekey=8453
2. http://www.medicinenet.com/cystinuria/article.htm
3. http://en.wikipedia.org/wiki/Diabetic_ketosis
4. http://en.wikipedia.org/wiki/Nitrite
5. http://www.traprockpeace.org/glen_lawrence_march04.html.
6. http://en.wikipedia.org/wiki/Urinalysis
7. http://en.wikipedia.org/wiki/pyrolurea
8. http://en.wikipedia.org/wiki/Urinary casts
9. http://en.wikipedia.org/wiki/Uric acids
10. http://www.livescience.com/humanbiology/fake_urine_040906.html.
11. http://www.webmd.com/hw/health_guide_atoz/hw5973.asp
12. http://en.wikipedia.org/wiki/Ketone
13. http://en.wikipedia.org/wiki/Nitrite
14. http://en.wikipedia.org/wiki/Proteinuria
15. http://en.wikipedia.org/wiki/Diabetes
16. http://etd.paml.com/etd/24hrreport.php
17. http://loudoun.nv.cc.va.us/vetonline/vet131/urinalysis.htm
18. http://en.wikipedia.org/wiki/Urobilinogen
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