AKARSANA KUMAR S11132245

BI309: COMPARATIVE ANIMAL PHYSIOLOGY

Tuesday 9-1 PM

KIDNEY PHYSIOLOGY AN D URINALYSIS

Organ Observations and Urine Test

Aim/ Objectives

- To examine and to note the gross structures of the kidney

- Draw the structure of internal and external gross structures and explain their functions
- Examine prepared slides of provided mammalian kidney, ureter and urinary bladder
- Examine urine for color, transparency, specific gravity
- To carry out the chemical test for abnormal urine constituents

Introduction

The kidneys are the chief regulators of the internal environment of the body. They do this by
regulating the concentration of ions, water, and pH in the various body fluids. In addition, they
provide for the elimination of the waste products of metabolism. The million nephrons in the
kidneys contain two main structures, the glomerulus and the renal tubule. As blood passes
through the kidneys, it is first filtered through the glomerulus (120 ml/min), and the filtrate then
passes into the renal tubule (Mason, 20120. The tubular filtrate is similar to blood plasma in
composition, except that large molecules(having molecular weights of more than 70,000) are
excluded (e.g., plasma proteins). As this filtrate passes along the proximal and distal tubules,
most of the water is reabsorbed, and many essential substances are actively or passively
reabsorbed into the bloodstream. Toxic by-products of metabolism and substances in excess are
retained in the filtrate or are secreted into the filtrate and finally excreted in the urine (1 ml
formed per minute). Thus, the final composition of the urine is quite different from that of the
glomerular filtrate and reflects the integrity of kidney function and changes in blood
composition. An analysis of urine can yield valuable information about the health of the kidney
and of the body in general. Various diseases are characterized by abnormal metabolism, which
causes abnormal by-products of metabolism to appear in the urine (Nigam, 2014).
 For example, phenylpyruvic acid appears in the urine in phenylketonuria (PKU), a disease
resulting in mental retardation. In diabetes mellitus, deficient production of insulin by the
pancreas results in the appearance of glucose in the urine (glycosuria). The volume of urine
produced and its specific gravity give information on the state of hydration or dehydration of
the body.
The purpose of this experiment was to analyze the provided non-human urine for pathological
constituents. The structure of the provided cattle and pig kidney was examined labeling the
gross internal and external structures.

Material & Method

-as per lab manual

Results and Discussion

Part A: Organ Observation

Parts of the Description and Functions

Kidney

Renal hilus An indentation near the center of the concavity of the kidney where the renal vein and
ureter leave the kidney and the renal artery enters the kidney.

Renal capsule A smooth, transparent membrane surrounding the kidney. It protects and helps maintain the
kidney's shape. It is also surrounded by fatty tissue which helps protect the kidney from
damage.

Renal cortex The outer reddish part of the kidney that has a smooth texture. It is where the Bowman's
Capsules, glomeruli, proximal and distal convoluted tubules and blood vessels are found.

Renal medulla The inner striated red-brown part of the kidney.

Renal Striped, triangular structures within the medulla, which are made of straight tubules and
pyramids corresponding blood vessels.

Renal pelvis The funnel-shaped cavity that receives urine drained from the nephrons through the
collecting ducts and papillary ducts.

Renal artery The blood vessel that delivers oxygen-rich blood to the kidney. It enters the kidney through
the hilus and divides into smaller arteries, which separate into afferent arterioles that serve
each of the nephrons.
Renal vein The blood vessel that receives deoxygenated blood from the kidney and returns it to the
systemic circulation.

Interlobular The blood vessel that delivers oxygen-rich blood to the glomerular capillaries under high
artery pressure.

Interlobular The blood vessel that receives low pressure deoxygenated blood that drains from the
vein glomeruli and the loops of Henle.

Kidney The functional units where the kidney's main functions are performed. There are about a
nephrons million nephrons in each kidney.

Collecting duct The part of the kidney nephron that collects urine and drains into papillary ducts, minor
calyx, and major calyx, and finally into the ureter and urinary bladder.

Ureter The structure which conveys urine from the kidney to the urinary bladder.

Part B: URINE EXAMINATION

I) Gross Examination

The physical characteristics of urine include observations and measurements of color, clarity,
odor, specific gravity and pH. Visual observation of a urine sample can give important clues as to
evidence of pathology.

Color

The color of normal urine is usually light yellow to amber. Generally, the greater the solute
volume, the deeper the color. The yellow color of urine is due to the presence of a yellow
pigment, urochrome. Deviations from normal color can be caused by certain drugs and various
vegetables such as carrots, beets, and rhubarb. Unusual or abnormal urine colors can also be the
result of a disease process. For an instance, red-colored urine can occur when blood is present in
the urine and can be an indicator of disease or damage to some part of the urinary system. The
depth of urine color is also a crude indicator of urine concentration. Pale yellow or colorless
urine indicates dilute urine where lots of water is being excreted. Dark yellow urine indicates
concentrated urine and the excretion of waste products in a smaller quantity of water, such as is
seen with the first morning urine, with dehydration, and during a fever (Madya 2012)

Odor

Urine odor refers to the scent of the fluid excreted during urination. Urine odor varies. Slightly
aromatic is characteristic of freshly voided urine. Urine becomes more ammonia-like upon
standing due to bacterial activity. Moreover, when a person is dehydrated, his/her urine will have
a smell similar to ammonia. However, if one drinks enough fluids and are otherwise healthy,
urine does not usually have a strong smell (Nigam, 2014). Changes in urine odor are usually
temporary. Such changes are not always a sign of disease. Certain foods and medicines,
including vitamins, may affect your urine's odor. For example, asparagus causes a characteristic
urine odor due to its content of asparagine, an amino acid. However, foul-smelling urine may be
due to bacteria, such as those responsible for urinary tract infections. Sweet-smelling urine may
be a sign of uncontrolled diabetes or a rare disease of metabolism. Liver disease and certain
metabolic disorders may cause musty-smelling urine (Dugdale, 2013).

pH

The kidneys play an important role in maintaining the acid-base balance of the body. Therefore,
any condition that produces acids or bases in the body such as acidosis or alkalosis, or the
ingestion of acidic or basic foods, can directly affect urine pH. The normal pH of urine ranges
from 4.5 - 8.0. Average is 6.0 which slightly acidic. Diet can be used to modify urine pH. A
high-protein diet or consuming cranberries will make the urine more acidic. A vegetarian diet, a
low-carbohydrate diet, or the ingestion of citrus fruits will tend to make the urine more alkaline.
Some of the substances dissolved in urine will precipitate out to form crystals when the urine is
acidic; others will form crystals when the urine is basic. If crystals form while the urine is being
produced in the kidneys, a kidney stone or “calculus” can develop. By modifying urine pH
through diet or medications, the formation of these crystals can be reduced or eliminated. In
addition, bacterial infections also increase alkalinity (Raven et al, 2015).

Specific Gravity

The specific gravity of urine is a measurement of the density of urine - the relative proportions
of dissolved solids in relationship to the total volume of the specimen. It reflects how
concentrated or dilute a sample may be. Water has a specific gravity of 1.000. Urine will always
have a value greater than 1.000 depending upon the amount of dissolved substances (salts,
minerals, etc.) that may be present. Very dilute urine has a low specific gravity value and very
concentrated urine has a high value. Specific gravity measures the ability of the kidneys to
concentrate or dilute urine depending on fluctuating conditions. Normal range 1.005 - 1.035,
average range 1.010 - 1.025. Low specific gravity is associated with conditions like diabetes
insipidus, excessive water intake, diuretic use or chronic renal failure. High specific gravity
levels are associated with diabetes mellitus, adrenal abnormalities or excessive water loss due to
vomiting, diarrhea or kidney inflammation. A specific gravity that never varies is indicative of
severe renal failure (Raven et al, 2015).

ii) Chemical Test for Abnormal Urine Constituents

The use of chemical methods for the analysis of urine allows for the discernment of its contents
through chemical reactions. Also, this method allows us to detect materials in the urine that
cannot be detected solely by the senses.

Glucose is not normally present in the urine because it is a renal threshold substance. This means
that it is not excreted in the urine until it reaches certain elevated levels in the blood. Glucosuria
or glycosuria is the term used for presence of glucose in the urine (U.S. National Library of
Medicine, 2012). To detect glucose, Benedict’s test was employed. The red precipitate often
appears orange since it is obscured by the blue color of Benedict’s reagent. Nevertheless, this
would still indicate a positive result. The test solution must be heated as the reaction is favored
with the presence of heat. The presence of glucose in the urine may be indicative of certain
diseases. One of these could be diabetes, which is caused by low quality or quantity of insulin.
However, further tests should be employed to confirm this disease. This may also be an indicator
for pregnancy since gestational diabetes may occur. A rare condition called renal glycosuria may
also be possible. This happens when glucose is excreted in the urine even if glucose is at normal
levels in the blood (U.S. National Library of Medicine, 2012).

Proteinuria is the presence of excess proteins in the urine. Temporarily high level of protein in
the urine after exercising, during pregnancy or during an illness is normal, but generally, there
should only be small traces of protein in the urine of a healthy individual (Healthwise, 2012). In
the experiment, a test using nitric acid and heat was performed for the detection of the protein
albumin in the urine. It is based on the principle of heat coagulation and precipitation of protein
by the nitric acid resulting to a cloudy solution or presence of white precipitate. It is highly
sensitive and can react with many proteins including albumin, globulins, glycoprotein, and
Bence-Jones protein (Byers, 2012). The presence of heat and strong acid causes the protein to
denaturize and coagulate. This disrupts the hydrogen bonds of the protein structure leaving only
its primary structure. It also changes the solubility of the protein wherein the water gets trapped
in a white semi-solid gel (Raven et.al, 2015).

The peroxidase activity of hemoglobin present in urine decomposes hydrogen peroxide and the
liberated oxygen oxidized benzidine to form a green- blue colored complex. Presence of more
number of red blood cells in urine is called hematuria which is associated with disease of or
damage to the genitourinary tract. Other disorder commonly used associated with hematuria
includes acute infection chronic glomerulonephritis tuberculosis of kidney nephritic syndrome
toxic damage to glomerulus malignant hypertension infarction renal calculi trauma to kidney,
acute cystitis, calculi, tumors in the ureter or bladder and kidney stones. In other clinical
conditions such as bleeding disorder (leukemia, thrombocytopenia, coagulation factor deficiency,
sickle disease or traits, scurvy), use of anticoagulant drugs.

Bile salts when present lower the surface tension of urine. When sulphur powder is added to the
urine, sulphur particles sink to the bottom of the tube. In the case of normal urine, it will float
on the surface. Bile salts along with bilirubin can be detected in urine in cases of obstructive
jaundice. In obstructive jaundice, bile salts and conjugated bilirubin regurgitate into blood from
biliary canaliculi (due to increased intrabiliary pressure) and are excreted in urine. The test used
for their detection is Hay’s surface tension test. The property of bile salts to lower the surface
tension is utilized in this test (Byers, 2012).

Nitroprusside used in this test reacts with both acetone and acetoacetic acid in presence of alkali
(NH4OH) to produce permanganate calomel red ring at the junction. In diabetes mellitus
impaired ability to metabolize carbohydrate takes place. as carbohydrate cannot be used to meet
the body energy need, fats are burned which leads to the presence of ketones in the urine.
Individuals receiving levadopa paraldehyde pyridium and phathalein compound may produce
false positive result when tested for ketonuria. Presence of salicylates gives false negative result
(Healthwise, 2012). Presence of ketones indicates increase in:

 Diabetes mellitus
 Propanol poisoning
 Severe starvation.
 Severe carbohydrate restriction
 Anorexia
 Fasting
 Fever
 Prolonged vomiting

ii) Microscopic Examination

The microscopic elements of the urine sediment are examined to detect and determine possible
renal and urinary tract disorders, including other systemic diseases. The microscopic examination
of urine sediment is done through centrifugation. The sediment contains red blood cells, white
blood cells, epithelial cells, casts, crystals and other structures, which may have accumulated in
the urine in its passage from the kidney to the lower urinary tract.
Red blood cells or erythrocytes (RBCs) have a normal (pale yellow) appearance. White blood
cells or leukocytes (WBCs) can be a sign of infection of other diseases in the kidney. They can
enter anywhere to the urine from the glomerulus to the urethra. Few renal epithelial cells can be
seen in urine microscopic examination due to normal exfoliation. Renal cells are the cells lining
the nephron, glomerulus, proximal and convoluted tubules and the collecting ducts. They are
indicative of renal diseases such as acute tubular necrosis, viral infections or renal transplant
rejection. Casts are solidified proteins formed in the lumen of kidney tubules. These are formed
in the distal convoluted tubules and the collecting duct of the kidney. Presence of casts in
microscopic examinations indicates existence of a renal disease.

Abstract

Abstract

Madya,(2012) performed uranalysis test of human urine and found out that Ketones (acetone,
aceotacetic acid, beta-hydroxybutyric acid) resulting fromeither diabetic ketosis or some other
form of calorie deprivation (starvation), are easilydetected using either dipsticks or test tablets
containing sodium nitroprusside.

Hathway(2012) carried out chemical analysis of ketone.The presence of abnormally high numbers of
ketones in the urine (ketonuria)usually results from uncontrolled diabetes mellitus. Ketonuria
can also be caused byprolonged diarrhoea or vomiting that results in starvation.

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