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Genitourinary

System
Laboratory Manual

3rd Year Undergraduate Program


Faculty of Medicine Universitas Padjadjaran
3/20/2015

TABLE OF CONTENTS
WEEK

DEPARTMENT

I
II
III
IV

Embryology
Anatomy
Histology
Pathology
Anatomy
Physiology

TOPIC

Embryology of GUS
Anatomy of GUS
Histology of GUS
Pathology anatomy of
GUS
Renal Regulation of
Fluid Balance
Clinical
Urine
Microscopic
Pathology
Examination
Microbiology Microbiology of GUS
Pharmacology Drugs Acting on the
kidney;
comparison
between some diuretics
agents

V
VI
VII
VIII

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

PAGE
3
11
16
39
58
62
68

79

WEEK I

EMBRYOLOGY OF GENITOURINARY SYSTEM


LEARNING OBJECTIVES

After performing laboratory activity, the student should be able to:


1.
2.
3.
4.

Describe development of kidney system.


Describe molecular regulation of kidney development.
Describe abnormalities/anomalies of the kidney development.
Describe the development and abnormalities of ureter, urinary bladder, urethra and
prostate.
5. Describe development of the indefferent embryo.
6. Describe primordium of the external genitalia.
7. Describe clinical considerations of the development of external genitalia.

RESOURCE PERSON

1. Retno Ekowati, dr., M.Kes

REFERENCES

1. Sadler, T.W. Langmans Medical Embryology. 10th Ed. Lippincott Williams and
Wilkins. Philadelphia. 2006. pp. 229-256.
2. Moore, K.L., and Persaud, T.V.N. The Developing Human. Clinically Oriented
Embryology. 7th Ed. Elsevier Saunders. 2003. pp. 255-285.

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

HOMEWORK ASSIGNMENTS
Complete the figure by naming the pointed parts:

Fig. 2.1 Transverse sections through embryos at various stages of development showing
formation of nephric tubules. A. 21 days. B. 25 days. Note formation of external and internal
glomeruli and the open connection between the intraembryonic cavity and the nephric tubule.

Fig. 2.2 A. Relationship of the intermediate mesoderm of the pronephric, mesonephric, and
metanephric systems. In cervical and upper thoracic regions, intermediate mesoderm is
segmented; in lower thoracic, lumbar, and sacral regions it forms a solid, unsegmented mass
of tissue, the nephrogenic cord. Note the longitudinal collecting duct, formed initially by the
pronephros but later by the mesonephros. B. Excretory tubules of the pronephric and
mesonephric systems in a 5-week embryo.

Genitourinary System 3rd Year Undergraduate Program


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Fig. 2.3 A. Transverse section through the urogenital ridge in the lower thoracic region of a 5week embryo showing formation of an excretory tubule of the mesonephric system. Note
the appearance of Bowmans capsule and the gonadal ridge. The mesonephros and gonad
are attached to the posterior abdominal wall by a broad urogenital mesentery. B. Relation of
the gonad and the mesonephros. Note the size of the mesonephros. The mesonephric duct
(wolffian duct) runs along the lateral side of the mesonephros.

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

Fig. 2.4 Relation of the hindgut and cloaca at the end of the fifth week.
The ureteric bud penetrates the metanephric mesoderm (blastema).

Fig. 2.5 A-C Ascent of the kidney. Note the change in position between the
mesonephric and metanephric systems. The mesonephric system degenerates
almost entirely, and only a few remnants persist in close contact with thr gonad.
In both male and female embryos, the gonads descend from their original level to
a much lower position.

LABORATORY ACTIVITY
1.
2.
3.
4.
5.
6.
7.
8.
9.

Discuss homework materials in small group, and explain development of the kidney.
Explain molecular regulation of the kidney development.
Explain clinical correlation that appear, if any, in the development of kidney.
Describe the Anomalies of Kidney.
Describe: The development of Ureter and Anomalies of the Ureter.
Describe: The development of Urinary bladder and Anomalies of Urinary Bladder)
Describe: The development of Male Urethra and Anomalies of Urethra
Describe: The development of the Prostate
Describe: The development of Male Genital Externa and Anomalies of Male External
Genitalia
10. Describe: The development of Testis and Descent of Testis

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

Complete the figure by naming the pointed parts:

Fig. 2.6 Development of the renal pelvis, calyces, and collecting tubules of the
metanephros. A. 6 weeks, B. At the end of the sixth week, C. 7 weeks, D. Newborn. Note
the pyramid form of the collecting tubules entering the minor calyx.

Fig. 2.7 Development of a metanephric excretory unit. Arrows, the place where the excretory unit
(blue) estblishes an open communication with the collecting system (yellow), allowing flow of urine
from the glomerulus into the collecting ducts.

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

Fig. 2.8 Genes involved in differentiation of the kidney. A. WT1, expressed by the mesenchyme, enables
this tissue to respond to induction by the ureteric bud. GDNF and HGF, also produced by mesenchyme,
interact through their receptors, RET and MET, respectively, in the ureteric bud epithelium, to stimulate
growth of the bud and maintain the interactions. The growth factors FGF2 and BMP7 stimulate
proliferation of the mesenchyme and maintain WT1 expression. B. PAX2 and WNT4, produced by the
ureteric bud, cause the mesenchyme to epithelialize in preparation for excretory tubule differentiation.
Laminin and type IV collagen form a basement membrane for the epithelial cells.

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

Fig. 2.9 A, B. A complete and a partial double ureter. C. Possible sites of


ectopic ureteral openings in the vagina, urethra, and vestibule. D, E.
Photomicrographs of complete and partial duplications of the ureters (U).
Arrows, duplicated hilum; B, bladder; K, kidneys; ML, median umbilical
ligament.

Fig. 2.10 A. Unilateral pelvic kidney showing the


position of the adrenal gland on the affected side. B, C.
Drawing and photomicrograph, respectively, of
horseshoe kidneys showing the position of the inferior
mesenteric artery. BW, bladder wall; U, ureters.

Genitourinary System 3rd Year Undergraduate Program


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----

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WEEK II

ANATOMY OF GENITOURINARY SYSTEM


LEARNING OBJECTIVES

1. Describe the anatomic topography of kidney.


2. Describe the relationship of kidney and suprarenal gland to adipose, fascial
coverings, and other abdominal organs.
3. Describe the vascularization, innervations and lymphatic vessels of kidney.
4. Describe basic structure gross anatomy of kidney.
5. Describe the branches of abdominal aorta and tributaries of inferior vena cava (level
of vertebrae).
6. Describe the anatomic topography of pelvic urinary organs.
7. Describe the anatomical aspects of ureters, urinary bladder and urethra.
8. Describe the vascularization, innervations and lymphatic vessels of ureters, urinary
bladder and urethra.
9. Describe the structure and function of ureters, urinary bladder, and urethra.
10. Compare the urethras of males and females.
11. Describe the anatomic topography of male and female genital organs.
12. Describe the anatomical aspects of male and female genital organs.
13. Describe the vascularization, innervations and lymphatic vessels of male and female
genital organs.
SEQUENCE

1. Pretest 15 minutes
2. Lab activity 120 minutes
a. Video guide
b. Specimen identification
c. Anatomy pictionary quiz
3. Post test 15 minutes

RESOURCE PERSON

1. Putri Halleyana A. R., dr


2. Gita Tiara D. N., dr
3. Nani M. Yazid., drg., Mkes

REFERENCES

1. Moore, KL and Dalley, AF. Clinical Oriented Anatomy. 5th Ed. Lippincott Williams &
Wilkins. 2006. pp. 308-320.

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NOTE

Before starting the lab activity, the student must:


1. Wear the lab coat properly
2. Bring the reference book or atlas
3. Finish their homework assignment
INTRODUCTION

Functionally, the urogenital system can be divided into two entirely different
components: the urinary system and the genital system.
The urinary system consists of two kidneys, two ureters, one urinary bladder, and
one urethrae. The superior urinary organs (kidneys and ureters) and their vessels are
primary retroperitoneal structures on the posterior abdominal wall.
The kidneys do the major work of the urinary system. They produce urine that is
conveyed by the ureters to the urinary bladder in the pelvis.

HOMEWORK ASSIGNMENTS

1. Please make a drawing of urinary system consist of both kidneys (and its attachment
to the IVC and abdominal aorta), both ureters, urinary bladder, and the urethra!
2. Describe the kidneys:
a. Topography
b. Vascularization. Make a schematic drawing of kidneys arteries & veins
c. Innervation
3. Describe the ureters anatomical course. Including the structures it crosses and
consctrictions
4. Describe the difference of the right and left suprarenal gland
5. Describe the urinary bladders:
a. Surface and borders
b. The difference of organ relations in male and female
c. Vascularization
d. Innervation
6. Compare the male and female urethra in a table diagram!
7. Make a drawing of the internal and external male genitalia! Explain the functions of
the organ briefly in the drawing!
8. Describe the topography, organ relations, vascularization, and innervation of:
a. Female internal genitalia organ
b. Female external genitalia organ

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LABORATORY ACTIVITY
I.

Video Introduction

II.

Identification
Structure

check

Urinary System
Kidney
Location
Superior & inferior pole
Topography: right & left kidney
Coverings
Fibrous capsule
Perinephric fat
Renal fascia
Paranephric fat
Surfaces: anterior & posterior
Internal structures
Renal Pyramid
Renal Column
Renal Papilla
Renal Pelvis
Major calices
Minor calices
Vascularization
Renal artery & its branching: right & left
difference
Renal veins & its tributaries: right & left
difference
Lymphatic
Innervation
Ureter
Location & course: abdominal part, pelvic part
Constrictions
Vascularization
Arterial
Veins
Lymphatic
innervation
Urinary Bladder
Shape & topography: empty vs filled, male vs
female
Fixation
External structure: apex, base, body, neck
Surfaces: superior, posterior, infero-lateral

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comment

Internal structure: ureter orifices, urethral orifice,


trigone, detrussor muscle
Vascularization
Innervation
Urethra
Male: prostatic part, intermediate part, spongy part
Female: course
Sphincter
Vascularization
Innervation
Genitalia
Male
Internal: topography, course, vascularization,
innervation
Testes
Epididymides
Ductus deferentes
Seminal vesicles
Ejaculatory ducts
Prostate: topography, organs relation, parts
surfaces, ducts
Bulbourethral gland
Spermatic cord
External
Scrotum
Penis
External structure
Internal structure
Muscles & ligaments
Vascularization
Innervation
Female
Internal: topography, course, vascularization,
innervation
Vagina
Uterus
Uterine tubes
Ovaries
External: topography, course, vascularization,
innervation
Mons pubis
Labia majora
Labia minora
Clitoris
Bulbs of vestibule

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Greater vestibular glands

Trainers authorization

III.

Anatomy pictionary quiz


Each group will be ask to hand 20 pictionary questions to be guessed by the other
group. The number of guessed question will determine the winner and loser. The
winning group will be given a privilege while the losing group will be given extra
anatomy assignment.

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Genitourinary System 3rd Year Undergraduate Program


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WEEK III

HISTOLOGY OF GENITOURINARY SYSTEM


LEARNING OBJECTIVES

After performing laboratory activity, the student should be able to:


1. List the organs of the urinary system and male genital system
2. Identify the structures and regions of a kidney and describe their organization
3. Describe the structure, function, and location of each component of a nephron
4. Describe the function of juxtaglomerular apparatus and identify its component
5. Trace the flow of blood through the kidney
6. Identify the components of the glomerular filtration barrier in a diagram of a portion
of a renal corpuscle
7. Describe the structure and function of ureters
8. Describe the structure and function of urinary bladder
9. Describe the structure of male urethra
10. Describe the location of the glands, ducts, and external genitalia of the male
reproductive system
11. Describe the general organization of the testis
12. Describe the differences in wall structure of excretory genital ducts.
13. Compare the prostate, seminal vesicles, and bulbourethral glands in terms of general
organization, epithelial lining, secretory products, and the point(s) at which their
secretions enter the excretory pathway.

RESOURCE PERSON

Nursiah Nasution, dr. (Department of Histology)

REFERENCES

1. Junqueira, L.Carlos. Basic Histology Text and Atlas. 12th Edition. Lange Medical Books
McGraw-Hill.
2. Young, B and Heath, J.W.Weathers Functional Histology. 4th Edition. Churchill
Livingstone.
3. Paulsen, Douglas F. Basic Histology Examination & Board Review. 2nd Edition.
Appleton & Lange.
4. Gartner, Leslie P and Hiatt, James L. Color Textbook of Histology. 2nd Edition. Saunders

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INTRODUCTION

The urinary system includes the kidneys and the urinary tract. Kidneys are paired
bean-shaped, retroperitoneal organs are located in the posterior wall of the abdominal
cavity. The components of urinary tract are ureters, urinary bladder, and urethra. The lumen
of the tract is characteristically lined by transitional epithelium, except for certain portions
of the urethra. The kidneys filter metabolic wastes and foreign substances from the blood;
regulate the ion, salt, and water concentrations of the fluids that bathe the bodys tissues;
and produce rennin and erythropoietin. The collection of raw filtrate from the blood in the
glomerular capillaries is only the first step in urine production. It is followed by the
reabsorption of important ions, small proteins, nutrients, and much of the water. These are
returned to the blood in the peritubular capillaries and vasa recta in precise proportions.
The portion of the raw filtrate that is not reabsorbed constitutes the urine; it is carried by
the ureters from the kidneys to the urinary bladder, where it is temporarily stored and later
released through the urethra.
The male genital system consists of the external genitalia and a series of glands and
ducts that produce and transport the male gametes (spermatozoa) and the seminal fluid.
During laboratory activity, the students will be asked to show any anatomical parts
using model, interactive CD, as well as histological preparations and finally also discuss
about important aspects of physiology and pathogenesis. Before activity, the students have
to accomplish the homework assignment first, read the primary references, so that they will
be more readily performing laboratory activity
The bladder and the urinary passages (calyces and renal pelvis) store urine formed in
the kidneys and conduct it to exterior. The ureters pass through the wall of the bladder
obliquely, forming a valve that prevents the backflow of urine. The urethra is a tube that
carries the urine from the bladder to the exterior. In men, sperm also pass through it during
ejaculation. In women, the urethra is exclusively a urinary organ.
The male reproductive system may be divided into four major functional components:
The testes or male gonads, paired organs lying in the scrotal sac, are responsible for
population of the male gametes, spermatozoa, and secretion of male sex hormones,
principally testosterone.
A paired system of ducts, each consisting of ductuli efferentes, epididymis, ductus
deferens and ejaculatory duct, collect, store and conduct spermatozoa from each testis. The
ejaculatory ducts converge on the urethra from which spermatozoa are expelled into the
female reproductive tract during copulation.
Two exocrine glands, the paired seminal vesicle and the single prostate gland, secrete a
nutritive and lubricating fluid medium called seminal fluid in which spermatozoa are
conveyed to the female reproductive tract. Semen, the fluid expelled during ejaculation,
consists of seminal fluid and spermatozoa, plus some desquamated duct lining cells.
The penis is the organ of copulation. A pair of small accessory glands, bulbourethral
glands of Cowper, secreted a fluid which lubricated the urethra for passage of semen during
ejaculation
The accessory genital glands produce secretions that are essential for the reproductive
function in men.
The prostate is a collection of 30-50 branched tubuloalveolar glands. The prostate has 3
distinct zones. The transition zone is of medical importance because it is the site where
most benign prostatic hyperplasia originates

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The seminal vesicles consist of 2 highly tortuous tubes about 15 cm in length they are glands
produce a viscid, yellowish secretion that contains spermatozoa-activating substances.
The bulbourethral glands, is 3-5 mm in diameter. The secreted mucus is clear and acts as a
lubricant.
HOMEWORK ASSIGNMENT

Please complete this figure by naming the pointed parts!

KIDNEY

organization
the kidney
(frontal
section)
Pyelum, ureter, renal artery,General
medullary
pyramid,ofcapsule,
cortex,
renal
vein, calyx mayor, calyx
minor

18

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

Please complete this figure by naming the pointed parts of renal vasculature!

Figure: Basic organization of the nephron, collecting system and renal vasculature
Glomerulus, afferent arteriole, efferent arteriole, cortical radial artery. Interlobar artery, arcuate
artery

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Collecting duct, proximal convoluted tubule, distal convoluted tubule, Henles loop

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

JUXTAGLOMERULAR APPARATUS

Diagram: Juxtaglomerular apparatus

Glomerulus, Bowmans space, Bowmans capsule, proximal convoluted tubule, distal convoluted
tubule, afferent arteriole, efferent arteriole , macula densa juxtaglomerular cell, lacis cells

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Schematic representation of a glomerular capillary with the visceral layer


of Bowmans capsule (formed by podocytes)

Podocyte cell body, filtration slit, Primary process, secondary process (pedicel)

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Genitourinary System 3rd Year Undergraduate Program


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Please list components of the wall structure of this figure!

Figure No.1: Transverse section of the ureter, Massons trichrome


Please list components of the wall structure of this figure!

Figure No.2: Section of wall layer the urinary bladder, Massons trichrome

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Dorsal
arteries

Superficial
dorsal vein
Deep dorsal
vein
Tunica
albuginea
Deep artery

Corpus
cavernosum
of the penis
Erectile
tissue

Urethra
Corpus
cavernosum
of the urethra

Drawing
Drawing of
of aa transverse
transverse section
section of
of the
the penis
penis
60

Figure No. 3

Urethra, corpus cavernosum of the urethra, corpus cavernosum of the penis, dorsal arteries,
superficial dorsal vein, deep artery, deep dorsal vein, tunica albuginea, erectile tissue

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Testis

testicular lobules, seminiferous tubules, septae, tunica albuginea, rete testis, tunica vaginalis
(parietal layer), tunica vaginalis (visceral layer), serous space, ductuli efferentes, ductus
deferens, epdidiymis

Figure: testis

25

Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

SEMINIFEROUS TUBULES, STRAIGHT TUBULES,


RETE TESTIS AND DUCTULI EFFERENTES

1. Seminiferous
tubules

2. Straight
tubules

3. Connective
tissue of
mediastinum
4. Rete testis
tubules
5. Ductuli
efferentes
(efferent
ductules
6. Rete testis
tubules

44

Rete testis tubules, ductull efferentes, rete testis tubule, connective tissue of mediatinum,
seminiferous tubules, straight tubules

26

Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Ductus
Ductus Epididymidis
Epididymidis (Duct
(Duct of
of the
the Epididymis)
Epididymis)
1. Connective
tissue
2. Cross sections
of the ductus
epididymidis

5. Section
through of Ubend of the
ductus
epididymidis

6. Epididymal
wall cut
tangentially
3. Basement
membrane

7. Smooth
muscle fibers
8. Stereocilia

4. Pseudostratified
columnar
epithelium with
stereocilia

9. Columnar cells
10. Basal cell

46

Basement membrane, connective tissue, cross sections of the ductus epididymidis,


pseudostratified columnar epithelium with stereocilia, basal cell, columnar cells,
epididymidis wall cut tangentially, stereocilia, section through of U-bend of the ductus
epididymidis

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts

Ductus
Ductus Deferens
Deferens (Transverse
(Transverse section)
section)
1. Outer
longitudinal
muscle layer

2. Circular
muscle layer
3. Inner
longitudinal
muscle layer

5. Lamina propria
6. Longitudinal crest
of lamina propria
7. Epithelium

8. Adipose tissue

4. Nerve and
blood vessels
in the
adventitious

49

Lamina propria, epithelium, adipose tissue, longitudinal crest of lamina propria, inner
longitudinal muscle layer, outer longitudinal muscle layer, circular muscle layer, nerve and
blood vessels in the adventitious

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Central zone, peripheral zone, transition zone


Please complete this figure by naming the pointed parts!

Prostate
Prostate Gland
Gland

1. Prostatic
concretions

4. Glandular
epithelium

5. Ducts
2. Glandular
alveoli

6. Smooth
muscle fibers

3. Glandular
ducts
7. Capillary and
venule

57

Glandular ducts, glandular alveoli, glandular epithelium, prostatic concretions, smooth


muscle fibers, capillary and venule.

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Prostate
Prostate Gland
Gland with
with Prostatic
Prostatic Urethra
Urethra
1. Prostatic
urethra
2. Colliculus
seminalis

5. Prostatic
glands (alveoli)

6. Smooth muscle of
the stroma
7. Dilatation of
the utriculus

3. Fibromuscular
stroma

4. Prostatic
glands (alveoli)

8. Diverticula of
urethral wall
9. Utriculus

10. Ejaculatory
ducts

57

Fibromuscular stroma, colliculus seminalis, prostatic glands (alveoli), prostatic urethra,


utriculus, ejaculatory ducts, diverticula of urethral wall, dilation of the utriculus, smooth
muscle of the stroma

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Genitourinary System 3rd Year Undergraduate Program


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Please complete this figure by naming the pointed parts!

Seminal
Seminal Vesicle
Vesicle

4. Glandular
epithelium
1. Crypts in the
mucosa

5. Primary fold in
the mucosa
6. Secondary folds

2. Muscular coat

7. Lamina propria

3. Adventitia

52

Lamina propria, primary fold in the mucosa, secondary folds, glandular


epithelium, crypts in the mucosa, adventitia, muscularis coat

Think first and answer the question correctly and concisely!


Identify the structures and regions of a kidney and describe their organization
1.
2.
3.
4.
5.

List the organs of urinary system and male genitale system


Describe the structure, function, and location of each component of a nephron
Describe the function of juxtaglomerular apparatus and identify its components
Trace the flow of blood through the kidney
Identify the components of the glomerular filtration barrier in a diagram of a portion
of a renal corpuscle
6. Describe the structure and function of ureters
7. Describe the structure and function of urinary bladder
8. Describe the structure of male urethra
9. Describe the location of the glands, ducts, and external genitalia of the male
reproductive system
10. Describe the general organization of the testis
11. Describe the differences in wall structure of excretory genital ducts.
12. Describe the microstructure of prostate gland
13. Describe the microstructure of seminal vesicles
14. Describe the microstructure of bulbourethral glands

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LABORATORY ACTIVITY

Pre-requisites: The students have to do the homework assignment and read the references
as listed in the first page.
Task:
1. Discus the homework materials in the small group (tutorial group).
2. See the specimen under microscope and try to make a schematic draw and put the
most important description based on the schematic draw.
Specimen No.1 (kidney, H.E.)
Schematic Draw ( cortex)

32

Description

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Specimen No.2 (kidney, H.E.)


Schematic Draw (medulla)

Description

Specimen No.3: ureter


Schematic Draw

33

Description

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Specimen No.4: urinary bladder


Schematic Draw

Description

Specimen No.4: transverse section of the penis


Schematic Draw

34

Description

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Specimen No.5: Seminiferous tubules


Schematic Draw

Description

Specimen No.6: Tubuli recti and rete testis


Schematic Draw

35

Description

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Specimen No.7: Ductuli efferentes

Schematic Draw

Description

Specimen No.8: Ductus epididymidis


Schematic Draw

36

Description

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Specimen No.9: Ductus deferens


Schematic Draw

Description

Specimen No.10: prostate gland


Schematic Draw

37

Description

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Specimen No.11: prostate gland with prostatic urethra


Schematic Draw

Description

Specimen No.12: seminal vesicles


Schematic Draw

38

Description

Genitourinary System 3rd Year Undergraduate Program


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WEEK IV

PATHOLOGY ANATOMY OF GENITOURINARY


SYSTEM
LEARNING OBJECTIVES

After performing laboratory activity, the students should be able to :


1. Explain the macroscopic and microscopic of post streptococcal glomerulonephritis
2. Explain the macroscopic and microscopicof chronic pyelonephritis.
3. Explain the macroscopic and microscopicof common kidney tumour
4. Explain the macroscopic and microscopicof polycystic kidney
5. Explain the macroscopic and microscopicy ofcystitis
6. Explain the macroscopic and microscopicof urothelial carcinoma of the bladder
7. Explain the macroscopic and microscopicof benign prostatic hyperplasia
8. Explain the macroscopic and microscopicof prostatic adenocarcinoma
9. Explain the macroscopic and microscopicof seminoma of the testes

RESOURCE PERSON

1. Hasrayati Agustina, dr.,SpPA


2. Hermin Aminah, dr,. SpPA

REFERENCE

Kumar V, Abbas AK and Fausto N. Robbins and Cotran Pathologic Basis of Diseases ( 2005),
7th Edition. Elsevier Saunders. pp .504-506, 960-1019

HOMEWORK ASSIGNMENT
A.

Fill in the blank box under each the picture with microscopic and macroscopic
appearance of each cases

LABORATORY ACTIVITY

Pre-requisites: The students have to do the homework assignment and read the references
as listed in the first page.
Task:
1. Discuss the homework materials in the small group (tutorial group).
2. See the specimen under microscope and try to make a schematic draw and put the
most important description based on the schematic draw.

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1. ACUTE POST STREPTOCOCCAL GLOMERULONEPHRITIS


A 10-year-old boy had just recovered from skin infection ( impetigo ).Two weeks
later, he developed malaise, nausea and slight fever , he passed dark brown urine.

Microscopic :

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2
3
4

1=

2=

4=

5=

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3=

2. CHRONIC PYELONEPHRITIS
A 40-year-old man presentedto the hospital because of recurrent urinary tract
infections. He complained back pain, fever, polyuria and his blood pressure was 170/100
mmHg. A renal sonogram revealed extensive scarring with pelvic and calyceal enlargement
and also cortical thinning.

Macroscopic :

Microscopic :

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3. WILMS TUMOR
A 3-year-old boy presented to the hospital because of a large abdominal mass. He
had hematuria, abdominal pain and constipation. USG examination revealed a large mass in
his left kidney.

Macroscopic :

Microscopic :

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4. RENAL CELL CARCINOMA


A 60-year-old man presented to the hospital because of flank pain and hematuria
since 1 month ago. He also complained fever, weakness and weight loss.

Macroscopic :

Microscopic :

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5. POLYCYSTIC KIDNEY
A 5 years old girl presented to the hospital because of recurrent urinary tract infection. USG
reveal cystic masses in both of her kidney

Macroscopic

Microscopic

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Schematic draw
Specimen No 2:

Specimen No 3:

Specimen No 4 :

Specimen No 5 :

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6. CHRONIC NON SPECIFIC CYSTITIS AND CYSTITIS CYSTICA


A 37 years old female came to the outpatient clinic because of disuria and frequency since 2 weeks ago .She
had no fever or flank pain

Macroscopic

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Microscopic:

7. UROTHELIAL CARCINOMA OF THE BLADDER


A 56-year-old male had several episodes of hematuria and dysuria in the past few week. The patient
was a chronic smoker for 30 years. A urinalysis show 4 (+) blood. The urine culture is negative. A cystoscopy
was performed, and a 2 cm, sessile, friable mass was seen on the left lateral wall.

Macroscopic:

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Microscopic :

8. BENIGN PROSTATIC HYPERPLASIA


A 65-year-old male had increasing difficulty in urination. He had to get up several
times at night because of a feeling of urgency. He has difficulty starting and stopping
urination. His serum PSA level was slighty increased.

Macroscopic:

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Microscopic :

9. PROSTATIC ADENOCARCINOMA
A 65-year-old male presented with a long history of urinary hesitancy nocturia.
Digital rectal examination revealed a hard, irregular prostate. Serum PSA level is 120 mg/ml.

Macroscopic :

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Microscopic:

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Specimen No 1:
Schematic Draw

52

Description

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Specimen No 2:
Schematic Draw

Description

Specimen No 3 :
Schematic Draw

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Description

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Specimen no 4 :

Schematic Draw

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Description

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10. SEMINOMA OF THE TESTES


A 57 year-old male had a unilateral enlargement of the testss. Physical exanination
showed enlargement of the left testes 5 cm in diameter

Macroscopic :

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Microscopic :

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Specimen No 1:
Schematic Draw

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Description

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WEEK V

PHYSIOLOGY OF GENITOURINARY SYSTEM:


RENAL REGULATION OF FLUID BALANCE
LEARNING OBJECTIVES
After performing laboratory activity, the students should be able to:
1. Describe the renal regulation of fluid balance.
2. Demonstrate and explain how the kidneys respond to the water loading and salt water
loading by analyzing changes in urinary volume, specific gravity and pH.
RESOURCE PERSON
1.
2.
3.
4.

Dr. med. Setiawan, dr. (Physiology)


Reni Farenia, dr., M.Kes (Physiology)
Yuni Lazuardi, dr., M.Kes (Physiology)
Nova Sylviana dr.M.Kes ( Physiology)

REFERENCES
1. Guyton AC and Hall JE. Textbook of Medical Physiology. 11th Edition. Elsevier
Saunders. pp 348-365.
INTRODUCTION
Human being can be fasting of foods for several days. But, this survival capacity is
very limited for water. Exposing to water challenges require the capability of regulatory
process which mostly involves the kidney ability. Urine volume, solute concentration, and
electrolyte content are adjusted by the kidneys to maintain homeostasis of the blood.
Drinking excess water or eating salty foods results in a rising blood volume, which is
followed by compensatory increases in the urinary excretion of the salt and water.
Diuretics are agents that increase the rate of urine formation (diuresis). These agents
are ion transport inhibitors that decrease the reabsorption of Na + at different sites in the
nephron. As a result, Na+ and other ions such as Cl- enter the urine in greater amounts
rather than normal, along with water, which is carried passively to maintain osmotic
equilibrium. The efficacy of the different classis of diuretics vary considerably, with the
increase in secretion of Na+ varying from less than 2% for the weak, potassium-sparing
diuretics, to over 20% for the potent loop diuretics. Their major clinical uses are in managing
disorders involving abnormal fluid retention (edema) or in treating hypertension where

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their diuretic action causes decreased blood volume, which leads to reduction in blood
pressure.
HOMEWORK ASSIGNMENT
1. How is the mechanism of water regulation reflected by urinary excretion once a person
is challenged with drinking excess water for instance in amount of 1 L?
2. What do you expect from the urinary excretion analysis (urinary volume, spesific gravity
and pH) in above condition?

LABORATORY ACTIVITY
Pre-requisites: The students have to do the homework assignment and read the references.
Task of Renal Regulation of Fluid Balance
Materials:
1. Urine collection cups
2. Urinometers
3. pH paper (pH range 39)
Procedure:
1. The students void their urine into collection cups at the beginning of the laboratory
session. In the analyses done in step 4, this sample will serve as thecontrol (time zero).
2. The students drink 1000 mL of water or salt containing water (4 gram of NaCl).
3. After drinking the solutions described in step 2, the students void their urine every 30
minutes for 2 hours. The urine samples are analyzed as described in step 4.
4. Each of the five urine samples collected are analyzed for volume, pH, and specific gravity,
as follows:
(a) Volume (mL). Measure the approximate volume of urine obtained and enter the data in
the table of the laboratory report.
(b) pH. Determine the pH of the urine samples by dipping a strip of pH paper into the urine
and matching the color developed with a colorchart. The urine normally has a pH
between 5.0 and 7.5.
(c) Specific gravity. Determine the specific gravity of the urine samples by floating a
urinometer in a cylinder (fig. 9.3) nearly filled with the specimen. Read the specific
gravity at the meniscus on the urinometer scale, making sure that the urinometer float

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is not touching the bottom or the sides of the cylinder. The specific gravity is directly
related to the amount of solutes in the urine and ranges from 1.010 to 1.025. (Pure
water should have a specific gravity of 1.000.)

Worksheet
Enter your data in the following table of observation:
Drinking of pure water
Time

Urine volume

Urine pH

0
30
60
90
120

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Urine Specific gravity

Drinking of salt containing water


Time

Urine volume

Urine pH

Urine Specific gravity

0
30
60
90
120

Application of the knowledge:


1.

How is the urine analysis results if a person is now challenged with the high salt
containing water?

2.

Could you explain the regulatory mechanism involved?

3.

What would be happened in a dehydrated person?

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WEEK VI

CLINICAL PATHOLOGY OF GENITOURINARY


SYSTEM: URINE MICROSCOPIC EXAMINATION
HOMEWORK ASSINGMENT
To be collected to your tutor at the day of lab activity

What will you do on your laboratory activity today?

I.

OBJECTIVE
At the end of the activity the students will understand and can describe about:
1. The recommended of preanalyticalstage of urine examination
2. Analytical stage: The urine microscopic examination
3. Post analytical stage of urine examination
4. The interpretation of the test results
5. Factors which can affect or interfere the results

II.

REFERENCE:
1. Strassinger SK; Di Lorenzo MS; Urinalysis and Body Fluids; 4 thed; Philadelphia;
F.A. Davis Company, 2001.

III.

INTRODUCTION
The results of urine microscopicexamination contains information about many of the
bodys major information can be obtained by inexpensive laboratory tests.

IV.

INTENDED USE
Reasons for performing the urinalysis include aiding in the diagnosis of disease,
screening asymptomatic populations for undetected disorders, and monitoring the
progress of disease and the effectiveness of therapy.

V.

EQUIPMENTS

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In this laboratory activity the students do the microscopic analysis of the urine by
light microscope. Another equipments: object glass, cover glass, glove, and tissue.
VI.

PROCEDURES
PREPARATION/PREANALYTIC PROCEDURE
1. Urine pot: Dry, clean, leak-proof (disposable pots are recommended).
2. Midstream urine:
- Patients must be provided with appropriate cleansing materials, sterile
pot and instructions for cleansing and voiding.
- Patients are instructed to wash their hands prior to beginning the
collection. Male patients should clean the glands beginning at the urethra
and withdrawing the foreskin. Female patients should separate the labia
and clean the urinary meatus and surrounding area.

ANALYTICAL PROCEDURE
A. PHYSICAL EXAMINATION
The physical examination of urine includes the determinations of the urine color,
clarity, and specific gravity. The results of the physical portion of urinalysis also
can be used to confirm of or to explain findings in the microscopic examination of
urinalysis.

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TEST PROCEDUREOF PHYSICAL EXAMINATION


Determination of color and clarity can do on urine in a clean, dry and leak-proof container.SG can
also determine with a refractometer.

This procedure must be followed exactly to achieve reliable results.


1. Use your gloves
2. Collect fresh, well-mixed, uncentrifuged urine specimen in a clean dry
container. Mix well immediately before using.

B. MICROSCOPIC EXAMINATION:
The microscopic examination of the urineuse a sediment of urine that had
centrifugated. Its purpose is to detect and to identify insoluble materials present
in the urine. The blood, kidney, lower genitourinary tract, external
contaminations all contribute formed elements to the urine. These include RBCs,
WBCs, epithelial cells, casts, bacteria, yeast, parasites, mucus, spermatozoa,
crystals, and artifacts. Examination of the urinary sediment must include both
identification and quantitation of the elements present.

Squamous epithelial cells

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Squamous epithelial cells

White blood cell and granular cast

Stained white blood cell cast

Mucus

Fatty cast

Uric acid crystals

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Triple phosphate crystals

Cystine crystals

Cholesterol crystals

POSTANALYTICAL: Recording and Reporting


MICROSCOPIC EXAMINATION
- RBCs: <1/hpf
- WBCs: < 6/hpf
- Epithelial cells: represent normal sloughing of old cell. Three types of epithel cells are
seen in urine: squamous, transitional, and renal tubular.
- Casts: negative
- Bacteria, yeast, parasites: negative
- Mucus: normal :a small amount. It has no clinical significance when present in either
female or male urine
- Spermatozoa: occasionally found in the urine of male
- Crystals: depends on the pH of urine; normal in acid urine: uric acid, acid urates, and
sodium urates; in alkaline urine: amorphous phosphate, triple phosphate, calcium
phosphate, calcium carbonate, and ammonium biurate. In abnormal urine you can
found: cystine, cholesterol, leucine, thyrocine, bilirubin, sulfonamides, ampicillin, and
radiographic dye.

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RESULT:

..
...
...
DISCUSSION:
..

..
..................................................................................
..............................................................
CONCLUSION:

.
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WEEK VII

MICROBIOLOGY OF GENITOURINARY SYSTEM


Topic : Laboratory examination of organisms associated with urinary tract and sexual
transmitted infections
I. General objective
After finishing this activity, the student will be able to:
Understood microbiological examinations to confirm the diagnosis of urinary tract and
sexual transmitted infections
II. Specific objective
At the end of laboratory practice, the student could:
1. Understood methods of specimen collection and the laboratory examinations to
confirm the diagnosis of urinary tract and sexual transmitted infections
2. Understood the methods of examination, isolation, and identification of bacteria
that cause infection of the urinary tract and sexual transmitted infections
Upon completion of this course, the student will be able to:
1. List examples of urinary and genital tract specimens.
2. Explain methods of urine sampling and genital specimen collection for microbiology
examination
3. Explain the use of the primary plating media for each specimen.
4. Describe urine culture technique for the diagnosis of UTI
5. Select incubation atmosphere, temperature, and time for each culture.
6. Interpret, and evaluate direct Gram stains of genital specimens.
7. Describe colonial morphology and growth characteristics of the bacteria
8. Determine appropriate biochemical tests or adjunct procedures required for
identification of significant isolates.
9. Interpretation antibiotic susceptibility tests as required and evaluate their
appropriateness with regard to the treatment
10. Explain microscopic examination for diagnose Treponema pallidum infection
11. Explain the procedure and principle laboratory diagnosis of Chlamydia trachomatis
infection : microscopic, direct antigen detection, PCR
12. Correlate culture results with clinical history and presentation.

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III. Methods

Presentation
Demonstration
Discussion

Laboratory Activity
The laboratory examination is an integral part of the Microbiology course. It provides the
student an opportunity to learn basic microbiological techniques, and introduces him/her to
a case based approach to microorganisms related to respiratory tract infection.
Steps in microbiology examinations:

Direct detect infectious agent


Visualization
Culture
Nucleic acid (DNA or RNA)
Antigen detection
Indirect detect hosts response to infection
Serological detect antibody

Direct detection:
o Visualization (Microscopic) - smears from lesions
o Light microscope and Gram stain, wet mount preparation
Bacteria
Fungal spores
o Electron microscope
Viruses
o Quick and easy if positive , negative results not definitive
Visualize bacteria:
secretion : genital discharge
mucosal smears : mucosal scraping
Direct detection:
Culture and identification
Growth medium : colony count, selective media
Adequate incubations conditions
Temperature
Atmosphere (O2, CO2)
Biochemical tests for identification
Expensive and time-consuming, positive results may be definitive
Inoculate enrichment or selective media, incubate 24 hours
Streak on agar plates, incubate 24 hrs
Pick presumptive colonies and restreak for identification

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METHODS OF URINE COLLECTION


1. Random collection taken at any time of day with no precautions regarding
contamination. The sample may be dilute, isotonic, or hypertonic and may contain
white cells, bacteria, and squamous epithelium as contaminants. In females, the
specimen may contain vaginal contaminants such as trichomonas, yeast, and during
menses, red cells.
2. Early morning collection of the sample before ingestion of any fluid. This is usually
hypertonic and reflects the ability of the kidney to concentrate urine during
dehydration which occurs overnight. If all fluid ingestion has been avoided since 6
p.m. the previous day, the specific gravity usually exceeds 1.022 in healthy
individuals.
3. Clean-catch, midstream urine specimen collected after cleansing the external
urethral meatus.
4. Catherization of the bladder through the urethra for urine collection is carried out
only in special circumstances, i.e., in a comatose or confused patient. This procedure
risks introducing infection and traumatizing the urethra and bladder, thus producing
iatrogenic infection or hematuria.
5. Suprapubic transabdominal needle aspiration of the bladder. When done under ideal
conditions, this provides the purest sampling of bladder urine. This is a good method
for infants and small children.
Method of clean-catch, midstream urine specimen collection:
1. Local disinfection of the meatus and adjacent mucosa should be performed with a
nonfoaming antiseptic solution; this region should then be dried with a sterile swab
to avoid mixture of the antiseptic with urine.
2. Contact of the urinary stream with the mucosa should be minimized by spreading
the labia in females and by pulling back the foreskin in uncircumcised males.
3. A midstream urine is one in which the first half of the bladder urine is discarded and
the collection vessel is introduced into the urinary stream to catch the last half. The
first voided specimen should be discarded since the initial urine flushes urethral
contaminants such as contaminating cells and microbes from the outer urethra. It is
the second, midstream sample that should be sent to the laboratory. This sounds
easy, but it isn't (try it yourself before criticizing the patient).
Urine collection of baby

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URINE CULTURE
1. QUALITATIVE URINE CULTURE
As previously discussed, the most common cause of UTI in men and women are gram
negative rods from the patients fecal flora; therefore, a qualitative approach in
demonstrating their presence should center around growth characteristics and biochemistry
associated with that group. The Qualitative isolation of urinary pathogens begins with
primary culture on Blood agar. Most all incriminating microorganisms will grow voraciously
on sheep blood with E. coli representing 80% of UTI, and many producing beta hemolytic
zones. Mac Conkey agar is utilized to further selectively isolate and differentiate those gram
negative rods that ferment lactose (e.g. E.coli, Klebsiella, Enterobacter and Serratia).
Patients who have high numbers of cells/ ml that grows on blood agar but not on
MacConkey s agar may suggest a more serious invasive pathogen of the descending route
(e.g. S. pyogenes , Candida or S. aureus).
CULTURE PROCEDURE
Step 1. Label the bottom of a blood agar and MacConkey agar plate Qualitative
Urine
Step 2. From the simulated urine, streak for isolation onto the blood and
MacConkeys agar.
Step 3. Incubate plates inverted at 370C for 24 hours.
Step 4. Record observations in results

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2. QUANTITATIVE URINE CULTURE


a. Pour plate method : isolation of bacteria by dilution techniques
b. Calibratied loop inoculation
A quantitative culture can be easily performed using a 1:1000 ml (0.001 ml) sterile,
disposable loop. The loop is dipped vertically into the mixed urine specimen and inoculated
onto a blood agar plate making a single line streak from the top of the plate to the bottom
of the plate. Distribution of the 1:1000 ml urine dilution is done by cross-streaking the initial
line of inoculation over the surface of the plate to obtain countable colonies. The plate is
incubated overnight at 350 C and examined the next day for countable colonies. The number
of colonies present is multiplied by 1000 (1000 represents the reciprocal of your dilution
factor RDF i.e. from the standardized loop 1/1000). Multiplying the number counted NC
by the (RDF) gives the number of organisms/ml of urine.
CFU/ml = NC x RDF
CULTURE PROCEDURE
Step. 1. Label the bottom of a blood agar plate Quantitative Urine
Step 2. Each desk is provided with 1/1000 ml standardized inoculating loops.
Aseptically remove one loop from the container
Step 3. Using the standardized loop, recover 1/1000 ml of urine from the
simulate specimen provided.
Step 4. Perform a quantitative culture streak
Step 5. Incubate plate at 370C for 24 hr.
Step 6. Record CFU/ml

c. Dip slide method


The functional unit consists of a clear plastic dual-chambered "paddle" containing both a
general purpose and a differential medium, Trypticase soy agar (TSA) and Levine eosin
methylene blue (EMB) agar or other kind of media, respectively. It is housed in a clear
plastic screw-cap vial. The media beds are parallel and face the same direction; each has a
surface size of about 310 mm2. The outer wall of the media wells is constructed to prevent
medium dislodgement during handling and shipment and the trapping of urine between the
wall and media. To inoculate the unit, the media portion is momentarily dipped into the
urine to wet the entire agar surfaces. It is then returned to the vial and incubated in an
upright position overnight at 370C. The number of colonies per milliliter of urine is estimated
by comparing the density of growth with a standard chart obtained.

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Microorganisms causing urinary tract infections


The bacteria that cause UTI will vary according to the clinical and epidemiological setting are
usually originating from the patients own normal intestinal organisms; colonic flora
(coliforms = Enterobacteriaceae) which, in certain circumstances, are able to pass from the
area around the anus into the urethra and so into the bladder. Members of this bacterial
group include opportunistic pathogens (e.g. E.coli by far the most frequent cause of urinary
tract infection), Klebsiella, Enterobacter, Serratia, and Proteus. Pseudomonas and
Enterococcus are also often incriminated, especially in hospitalized patients with indwelling
urinary catheters.
Community acquired UTI, otherwise healthy adults
Escherichia coli most common
Staphylococcus saprophyticus (usually young adult females)
Less commonly:
Proteus species
Enterobacter species
Serratia species
Morganella species
Klebsiella species
Citrobacter species
Pseudomonas aeruginosa
Enterococcus species
(Patients with recurrent UTIs, urinary tract abnormalities, obstruction and neurogenic
bladder will have a higher frequency of those bacteria listed as less common above).

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Hospital acquired UTI


Hospitalized patients rapidly become colonized by the hospitals resident microflora,
specifically Gram negative bacteria. Patients in this setting are prone to UTI because they
are debilitated and may have had urinary tract instrumentation, particularly urinary tract
catheterization. Indwelling urinary catheters universally become colonized by bacteria.
Hospital bacteria have been selected by broad spectrum antibiotics used in the hospital, and
are generally quite resistant to antibiotics.
Causes of hospital acquired UTIs include:
E. coli and Enterobacteriaceae as described above.
Increased frequency of Proteus, Enterobacter, Serratia, Morganella.
Pseudomonas aeruginosa and other Pseudomonas species.
Enterococcus faecalis and E. faecium.
Stenotrophomonas maltophilia
Corynebacterium urealyticum
Staphylococcus epidermidis
Candida albicans and other species.
Uncommon causes of UTI
Some other microorganisms which may infect the urinary tract should be mentioned:
Mycobacterium tuberculosis renal TB; uncommon in this community.
Adenovirus type 11 haemorrhagic cystitis in children.
SEXUALLY TRANSMITTED INFECTIONS
N. gonorrhoeae
Specimen collection : Dacron or rayon swab
Specimen transport
1. Direct inoculation ~ Candle jars
2. Nonnutritive transport media ~Amies medium (12 hrs) should not be refrigerated
3. Nutritive transport systems ~ Transgrow, JEMBEC, Bio-Bag, Gono-Pak
Transgrow Medium is a convenient flask containing MTM Chocolate agar and CO 2.
Culture
Selective Media for N. gonorrhoeae are Thayer-Martin (Modified=MTM), New York
City Medium (NYCM)
Incubation condition 35oC, 3-5% CO2
Colonial morphology : 5 types autolysin
N. gonorrhoeae forms small, convex, smooth, grayish-white to colorless, mucoid colonies in
48 hours at 35-370C.
The gonococcus requires: enriched medium with increased CO2 tension for growth
Cultured on modified Thayer Martin (MTM) Chocolate agar.

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MTM Chocolate agar is selective for pathogenic Neisseria contains :


enrichment factors to promote the growth of
gonococci,
antibiotics to inhibit normal body flora :
vancomycin to inhibit gram-positive bacteria;
colistin to inhibit gram-negative bacteria;
trimethoprim to suppress Proteus;
nystatin to inhibit yeast.
Identification tests
1. Oxidase test
tetramethyl--phenylenediamine dihydrochloride

Oxidase
indophenol (blue)

2. Carbohydrate utilization tests


Cystine trypticase agar (CTA) ~ 1% carbohydrate, phenol red
Organism
N. gonorrhoeae
N. meningitides
N. lactamica

Glucose
+
+
+

Maltose
+
+

Lactose
+

Sucrose
-

Chlamydia trachomatis
Chlamydia testing is available by 3 methods:
Culture : cell culture
Antigen detection from genital specimen: DFA. Elisa
Molecular technique: Molecular Probe, PCR
Chlamydia Culture Collection
1. Utilizing a sterile swab (rayon or dacron), obtain a suitable specimen.
2. Place the swab in transport media (M4 media tube with pink liquid).
3. Label the transport media vial.
4. Transport the vial to the laboratory. May be refrigerated.
Chlamydia DFA/ Elisa Collection
The Direct Fluorescent Antibody (DFA) method for Chlamydia utilizes an antibody to detect
Chlamydia organisms on a slide, while Elisa detect chlamydia antigen using microtiter plate.
This method can be used for urethral, cervical, and rectal samples.
Urethral samples (male):
Patient should not have urinated one hour prior to sampling.
1. Insert small dacron swab (mini-tip) 2-4 cm into urethra.
2. Rotate swab and withdraw.

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Cervical samples (swab):


1. Wipe exocervix with large or small rayon or dacron swab to remove all excess mucus.
Dispose of swab.
2. Gently insert rayon or dacron swabs into endocervical canal until most of dacron tip is not
visible.
3. Rotate swab 5-10 seconds inside endocervical canal.
4. Withdraw swab without touching any vaginal surfaces.

Rectal samples:
Samples should be collected only from symptomatic patients.
1. Insert the large dacron swab about 3 cm into anal canal.
2. Move swab from side to side to sample crypts.
3. Withdraw swab. If fecal contamination occurs, discard swab and obtain another
specimen.
Treponema pallidum
Laboratory Diagnosis: This relies heavily on clinical manifestations. In addition, the finding of
Treponema pallidum within exudative lesions by the use of dark-field microscopy and
positive serology aids the diagnosis.
Penicillin treatment eradicates all stages, including congenital infection in pregnancy.
PROCEDURE
Step 1. Observe Prepared Slide Treponema pallidum. Special stain or negative stain
(bright field microscope)
Step 2. Observe Dark Field Microscopy (presumptive Identification)
Step 3. Record results
TERMS AND QUESTIONS FOR STUDY
UTI exercise
1. What is bacteriuria?
2. How do microorganisms enter the urinary tract?
3. What group of microorganisms appear in ascending UTIs.
Neisseria gonorrhea exercise
1. Gram stain
2. Bacteria isolation, media
3. Colony morphology
4. Identification procedure
Spirochaeta exercise
1. Special stain
2. Differentiate morphology of:
Treponema pallidum
Borrelia bergdorferi
Leptospira interrogans

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ADDITIONAL QUESTION
1. What is the significance of small pin point, beta hemolytic, and colonies appearing
on blood agar but no growth on MacConkey agar when conducting a qualitative
urine culture?
2. What are significant numbers for a UTI when conducting a quantitative urine
culture? How many CFU/ml would you conclude the number of colonies if you find
145 colonies on the plate when you inoculate using a 1/1000 ml calibrated loop?
What number of colonies if you use a 1/100 ml calibrated loop?
3. Why is urine specimens keep cold until plated?
4. What do we learn from a dip stick test? Is this test conclusive?
5. Can we distinguish between N. gonorrhoeae and N. meningitidis by Gram stain?
6. What is the meaning of intracellular Gram negative cocci? Explain!
7. Why are selective media needed in primary isolation from female urogenital
specimens?
8. Can we diagnose syphilis from Gram stained smears of primary lesions? Explain!
9. Explain dark field microscopy and its application in the Microbiology laboratory!

DRAW THE RESULT OF YOUR ACTIVITY

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2014 - 2015

Signature of trainer

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Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

WEEK VIII

PHARMACOLOGY OF GENITOURINARY SYSTEM:


DRUGS ACTING ON THE KIDNEY; COMPARISON
BETWEEN SOME DIURETICS AGENTS
LEARNING OBJECTIVES

After performing laboratory activity, the students should be able to:


1. Explain pharmacological properties of diuretics related to the clinical applications

RESOURCE PERSON

1. Kuswinarti (Pharmacology)
2. Ike Husen (Pharmacology)

REFERENCES

1. Benowitz. N.L. Antihypertensive agents. Katzung B.G (editor). In: Basic and
Clinical Pharmacology. 10 th Edition. McGraw Hill LANGE. 2007: 159-179
2 Ives.H.E. Diuretic agents. Katzung B.G (editor). In: Basic and Clinical
Pharmacology. 10 th Edition. McGraw Hill LANGE. 2007: 236-252

INTRODUCTION

Diuretics are drugs that increase the excretion of Na+ and water from the body by an action
on the kidney. Their primary effect is to decrease the reabsorption of Na+ and Cl- from the
filtrate, increase water loss being secondary to the increased excretion of NaCl. This can be
achieved by a direct action on the cells of the nephron or indirectly modifying the content of
the filtrate. Since a very large proportion of the NaCl and water that passes into the tubule
in the glomerulus is reabsorbed, a small decrease in reabsorption can result in a marked
increase in excretion.
Before activity, the students have to accomplish the homework assignment first, read the
primary references, so that they will be more ready in performing laboratory activity.

MATERIALS, EQUIPMENTS AND EXPERIMENTAL SUBJECTS


MATERIALS:

79

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

a. Diuretic Agents
a. HCT
b. Furosemide
c. Spironolacton
b. Other :
d. Glycerin
e. Aquadest
EQUIPMENTS :
f.
g.
h.
i.
j.
k.

Beaker glass
Measuring glass
Catheter
Fixation board
Disposable injection
NGT

EXPERIMENTAL SUBJECTS :
Three months old healthy male rabbits with the same body weight
The rabbits are divided into 2 group :
a. Control group will receive aquadest only
b. Treatment group will receive diuretic agents consist of HCT/ Furosemid
/Spironolacton.

PROCEDURES
a.
b.
c.
d.

Fix the rabbit on the fixation board


Put the catheter into the ostium urethra externum
Emptying the vesica urinaria
Administer orally 15 cc of water to the first rabbit as the control and 15 cc of
diuretic agent to the other (each diuretic type for different rabbit).
e. Collect the urine after 30 minutes and repeat it twice every 30 minutes.
f. Compare the volume of urine from :
i. Control group vs treatment group
ii. Treatment group vs treatment group

80

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

RESULT

Volume (ml)
Group

Drug

before
treatment

Volume (ml) after treatment

30 minutes

60 minutes

II

III

IV

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Genitourinary System 3rd Year Undergraduate Program


2014 - 2015

90 minutes

120 minutes

DISCUSSION

CONCLUSION :

HOMEWORK ASSIGNMENT
A. DIURETICS

1. Which is the most potent diuretic found in your experiment? Give the rational
explanation of your experiment! (regarding either its mechanism of action or
pharmacokinetic)
2. List the indications of furosemide! Explain the rational explanation regarding its
mechanism of action!
3. Explain the side effects of furosemid
4. Explain the indication/s of HCT and its related mechanism of action!
5. Explain the mechanism of action of spironolacton
B. ANTI HYPERTENSIVE AGENTS

Explain the best drug of choice and worst choice for each patient below!
1. A-50-year old well controlled type 2 diabetes, male patient recently has been
diagnosed as stage II essential hypertension with normal renal function (no
microalbuminemia).
2. Hypertensive patient with tachycardia and chronic open angle glaucoma
3. Hypertensive patient with history of vasospatic angina pectoris
4. Stage II essential hypertension patient with pregnancy.
5. Patient who tends to have exacerbated asthma. Mention antihypertensive agent
should be avoided and explain the mechanism!
6. A patient has Stage III essential hypertension but normal ventricular function. After
evaluating the responses to several other antihypertensive agents, alone and in
combination, the physician places the patient on oral hydralazine. Mention
antihypertensive agent/s is/are likely to be needed to manage the expected and
unwanted cardiovascular side effects of hydralazine and explain!
C.NEPHROPHARMACOLOGY

1. What are guidelines to preventing drug-induced nephrotoxicity


What is drug-induced nephrotoxicity

82

Genitourinary System 3rd Year Undergraduate Program


2014 - 2015