1

Navayug vidyapeath Trust, Ladavali, Mahad, Raigad.

Project certificate
(Academic year 2021-22)

To whomsoever it is concerned

This is the certify that miss kudupkar Asiya Asif Roll No __________,
Enrollment No 2002250006 of our instituted studying in final semester
of the course Advanced Diploma in Medical Laboratory Technology
affiliated to the Maharashtra state board of technical education,
Mumbai, has satisfactory completed the project title urine Analysis &
urinary Tract Infection. Navayug vidaypeath trust ladavali as a part of
curricular requirement of the subject project and seminar on laboratory
training
This project represents her bonafide work during final semester of her
course.
This certificate is issued subject for certification of her training work by
the training institute as per the academic requirement of her laboratory
training project.

Professor incharge
Principal
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Project Title

Urinary Analysis & Urinary Tract Infection

at Ahamadiya clinical laboratory from
June 2021 to Nov 2021

Submitted in partial of the course

Advanced Diploma in Medical Laboratory Technology
Semester 3rd
Batch (2021-22)
Affiliated to the Maharashtra State Board
Of Technical Education, Mumbai
Institute name: - Navayug Vidyapeath Trust, Ladavli

Professor In charge (External) Signature of

candidate

Miss: Kudupkar Asiya Asif

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Dedicated

I dedicate this laboratory research project

to my beloved parents
My Teachers
&
My dear Friends
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Acknowledgments

I would like to express my sincere thanks to AHAMADIYA CLINICAL

LABORATORY for providing me the opportunity and resources to carry
out the project work. I would specifically thank Put owner name of
laboratory for being a constant source of motivation and support. I am
also thankful to him for providing me all the facilities and assistance for
carrying out the research work.

I owe my profound gratitude to our principal Mrs. Wadkar madam

(Navayug vidyapeath college) for her guidance and encouragement
during the entire period of the project.
I would like to acknowledge Mr Mansoor Lokhande (Lab Technician),
Mrs. Rupali Jadhav (B.Sc. DMLT). Foe their kind guidance and support.

I once again thanks to all of the staff members and others who have
directly or indirectly help me during my tenure.

Miss Kudupkar Asiya Asif

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Introduction
Urine Analysis contain, Quantity, Colour,
pH, Sugar & Protein is used for urine
report.

Urine Tract Infection is a bacterial

infection. When we are doing examine of
urine within 10 minutes and we saw
bacteria these bacteria known as
pathogenic bacteria is caused for disease,
in urine see calcium oxalate & urine acid
is symptom of calculla patient.
 6

Review of literature
a. The urinary system: - The mamaline urinary
tract is a contiguous hollow organa system
whose primary function is to collect, transport,
store and expel urine periodically and in highly
coordinated fashion. In so doing the urinary
tract ensure, the elimination of metabolic
products and toxic wastes generated in
kidney. The process of constant urine flow in
the upper urinary tract and intermittent
elimination from the lower urinary tract also
plays a crucially important part in cleansing
the urinary tract, ridding it of microbes that
might have already gained access when not
eliminating urine, the urinary tract acts
effectively as a closed systems inaccessible to
the microbes comprised, from proximal to
distal of renal papillae, renal pelvis, ureters,
urinary bladder, and urethra, each component
of the urinary tract has distinct anatomic
features and performs critical functions
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i. Kidney: - There is a pair of kidneys that

are purplish- brown and are located
below the ribs in the middle of the back.
Their function Is to

 Remove waste from the blood in the

form of urine
 Keep substances stable in the blood
 Make erythroprotein, a hormone
which helps to make red blood cells
 Make vitamins D-active
 Regulate blood pressure
The kidney removes waste from the
blood through tiny filtering units called
nephrons. Each nephron is made up of a
ball of small blood capillaries, called a
glomerulus, there is also a small tube
called a renal tubule which drain the
urine and joins other tubules carrying the
urine out of the kidney to the ureter,
urea, together with water and other
waste, forms the urine,
ii. Two ureters: - Each kidney has a narrow
tube called a ureter which carries urine
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from the kidney to the bladder. Muscle in the

ureter walls tighten and relax forcing urine
down this tube away from the kidney. If the
urine backs up or stand still a kidney infection
can develop. About every 10 to 15 Seconds.
Urine is emptied into the bladders from the
ureters.

iii. Bladder: - The bladder is a triangle shaped

hollow organ located in the lower abdomen. It
is held in place by ligaments attached to the
pelvis bones. The bladder walls relax and
expand to store urine and contract and flatten
to empty urine through the urethra

iv. Two sphineter muscles: - Circular muscles that

helps to keep urine from leaking by closing
tightly like a rubber band around the opening
of the bladder

v. Nerve and urethra: - The nerves alert a person

when it is time to urinate or empty the bladder
muscles to tighten, which squeezes urine out
of the bladder. At the same time the brain
signals the spineter muscles to relax to let
urine exit the bladder through the urethra.
When all the signals occur in the correct order,
normal urination occurs
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2. Urinary tract infection

Urinary tract infection (UTI) is a common bacterial infection

known to affect the different part of the urinary tract and the
occurrence found in the both male and female. Despite the fact
that both genders are susceptible to the infection women are
mostly vulnerable due to their anatomy and reproductive
physicology. The infection is usually caused as a consequence of
bacterial inversion of the urinary tract including the lower and the
upper urinary tract
Anatomy UTI’S are classified as lower UTI & upper UTI:
a. Upper UTI
 Involves kidney and ureter
 Acute pyelitis
 Acute pyelonephritis
b. Lower UTI
 Urethritis
 Cystitis
 Prostatitis in (male)
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3.1 Clinically
urinary tract infection is
classified as either asymptomatic or
symptomatic

Asymptomatic bacteriuria is defined as the

presence of significant bacteria without the
symptoms of an acute urinary tract
infection. Traditionally diagnostic criteria of
significant bacteriuria include culture of 105
colony formines units (CFUS) of a single
uropathogen in clean catch urine
spearmen’s. UTI is defined as the presence
of at least 105 organism per millimeter of
urine in an asylpto patient, or as a more
than 103 organism of urine with
accompanying pyuria (>7 white blood cell )
in a symptomatomatic patient symptomatic
urinary tract infection are divided into
lowest tract. (acute cystitis) or upper tract
(acute pyelonephris) infection cystitis is
defined as significant Bacteria with
associated Bladder mucosal invasion
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a. Predisposing factors for UTI

1. Age: - patient at old age are more

susceptible to urinary tract infection as
compared to young
2. Sex: - sexually active females (due to
short urethra proximity to anus trauma)
are more vulnerable to get UTI’s

3. Pregnancy: -
Because of anatomical and physiological
adaptation in pregnancy such as dialation
of ureter and renal pelvis, static,
incompetence of vesico urethral valves,
hormonal changes the risk of UTI
increases in pregnancy
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4.1 UTI in pregnancy: -

The prevalence of systematic and asymptomatic

bacteuria among women during pregnancy is very
common and previous history of the infections is a
major risk factor. It is a widely accepted fact that
pregnancy is associated with variety of anatomical
changes in a woman, followed by hormonal and
physical changes which increase the possibility of
urinary static which in turn cause the backward flow of
urine from bladders to ureters. This as a consequence
of physical aberrations enhances the occurrence of UTI
among women during pregnancy.
Untreated UTI or asymptomatic bacteriuria during
pregnancy may lead to serious consequence from
maternal life and fetus like higher risk of pyelonephritis
sepsis and transient renal failure and complicated
outcomes such as intrauterine growth restriction
preclampsia and premature delivery. Therefore, it is
important to screen, raise suspicion and known how to
recognize this condition. Intending to promptly initiate
appropriate treatment in order to minimize
complications associated with UTI.
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4. AIMS & OBJECTIVES

a. To find the incidence of asymptomatic

bacteria and symptomatic urinary tract
infection

b. To identify the isolated uropthogens by

using standard microbiological
techniques
,
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5. Material and Methods

The present study was prospective study carried out in

the department of microbiology Navayug vidyapeath trust
ladavali after approval from the institutional eithic
committee

6. Inclusion criteria

a. Normal patient urine

b. Exclusion criteria
i. Patients having a known renal disease/ anomaly
of urine tract
ii. Patient on antibiotic therapy within 72 hrs of
specimen collection

7. Investigation

a. Specimen collection:

i. Clean catch midstream urine specimen is

collected in a sterile wide mouth container
ii. Next portion of urine (midstream sample) is
collected in a sterile wide mouthed container
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b. Transport and storage:

i. Urine specimen are transported to the laboratory

without delay
ii. In case of delay, these samples are store in refrigerator
at 4 ᵒC for up to 4 hrs.

c. Processing of specimen:

i. Macroscopic examination:

1. Appearance of cloudy or turbid urine, this is seen

in presence of bacteria, proteins, crystals or
leucocytes (WBC’s casts)
2. Presence of blood (RBC’s), sometime when lady in
her period RBC’s known as non-pathogenic.
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ii. Urine culture:

1. Uncentrifuged urine specimen is cultured on blood

again and macconkey agar by semi quantitative
methods
2. A standard inoculating loop which holds 0.001 mL
of specimen is used and streaked on culture plates
without intermitted heating
3. Culture plates are incubated at 37 ᵒC for 24 hrs
and observed for growth of bacteria on next day
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iii. Identification of the isolate:

1. These isolated were identified using standard

microbiological techniques such as motility, gram
staining colony characteristics and biochemical
reaction

iv. Interpretation of results:

1. A colony count of 100 CFU per 0.001 mL which

corresponds to 105 CFU/mL of urine) at a single
uropathogenic bacterium on each culture plate is
considered as significant and is taken as a case of
urinary tract infection for both asymptomatic and
symptomatic patients
2. In a symptomatic patient with pyuria (>7 WBC’s P
or HPF in wet mount of urine) low bacterial counts
of 103 CFU/mL of urine are also considered
significant
3. The specimen (cultures) showing no growth of
bacteria or growth of < 102 CFU/mL of urine are
considered significant. They occur because of
contamination by commensal bacteria during
voiding
4. The frequency of intermediate 102 -103 CFU/mL is
only 5% to 10%. If the urine collection procedure
has been performed properly, in such a case fresh
specimen of urine is collected
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5. Urine specimen showing growth

of three or more type of
bacteria colonies are
considered as contamination
and a repeat specimen is
collected for such patients

6. In case of less frequent but

known urinary pathogen such
as staphylococcus aureus and
psedomonas aeuginosa low
colony counts are also
considered as significant
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8. Summary & Conclusion:

Urinary treat infection is a very common

infection among both man and women but
definitely a higher incidence in females
specially in pregnancy not only due to
anatomical differences between male and
females urinary tracts but also due to
physiological and hormonal changes
associated to serious consequence for
maternal life and fetus, transient renal failure
and complicated outcome such as intrauterine
growth restriction. It is important to screen
raise suspicion and know how to recognize
this condition intending to promptly initiate
appropriate treatment in order to minimize
complications associated with UTI
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9. Formation of urine:

The urine form urine passes through the uraters to the bladder for
excretion. The composition of urine reflects the activities of the
nephrons in the maintence of homeostasis. There are three
phases on the formation of urine
a. Simple filtration
b. Selective reabsorption
c. Secretion

a. Simple filtration
A large volume of blood approximately onelitre/ minute
flow through the kidney. In four to five minutes a volume of
blood equal to the total blood volume passes through the
renal circulation
A net filtration pressure of about 15 mmHg in the capillary
bed of the tuft drives the filterates through the glomerular
membrane. The filtrate is called an ultrafilter because its
composition is essentially the same as that of plasma.
However, ultrafilter does not contain most of the proteins
present in plasma having molecular.
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Weight greater than 15000 and also the blood cells. Each
nephrons produces about 100 mL of ultrafilter per day.
Since each kidney contain about one million nephrons,
about 170 to 200 liter of ultrafilter is produced per day and
per minute about 125mL pf filterate is formed. i.e.
glomerular filteration rater (GFR) is mL/minute, chemically
glomeluar filterate is an essential protein-free extracellular
fluis or a protein and cell free filterate whole blood.

b. Selective Reabsorption
The glomerular filtrate contains various substances such as
high threshold substances (glucose, amino acids, protein,
etc.) and low threshold substances (sodium, potassium,
calcium, etc.) high threshold substances are reabsorbed
almost completely by the tubuler, when their concentration
in the plasma are within the normal range but are not
completely reabsorbed.
Nearly 60 % to 70 % of ultrafilter is reabsorbed in the
proximal part of the tubule. The other substances
reabsorbed in the proximal part of the tubule are sodium
ions without water are reabsorbed generating diluted urine.
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Water reabsorption in the distal tubules and collecting

ducts is then regulated by antidiyretic hormone (ADH,
secreted by pituitary gland).
c. Secretion in the distal tubule

Some substances are actively transported (or actively

excreted) into the tubular lumen. The substances secreted
by the tubule epithelium are creatinine and potassium,
similarly H+, Nh4+ ions are also excreted and reabsorption to
Na+ & HCO3+ takes place in this region.

Out of 170 liters of glomecular filtrate produced in a day the

tubules reabsorb about 108.5 liters of the filtrate about 1.5
liters of urine is excreted in 24hr, which contains 25-30 g
urea 2-3 g creatine, 1-1.5 g uric acid, 1.5 g of sodium
chloride.
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10.1Acidic urine:

The American association of chemical chemistry says the normal

urine pH is between 4.5 to 8 any pH higher than 8 is basic or alkaline
and any under 6 is acidic. A urine pH test is carried out as a part of a
urinalysis. After performing urine pH test doctors can use the results
to diagnose various diseases. The kidney maintains normal acid-
base balance primarily through the reabsorption od sodium and the
tubular secretion of hydrogen and ammonium ions. Urine becomes
increasingly acidic as the amount of sodium and excess acid
retained by the body increases.
Acidic urine can also create an environment where kidney stones.
Stones can form if a person has low urine pH, meaning that it is
more acidic. It might indicate a medical condition such as, diabetic,
ketoacidosis, which is complication of diabetic, diarrhea stairation
uric acid crystals invariably form in acidic urine, typically with a urine
pH < 5.5. uric acid is soluble in alkaline urine, presenting the
precipitation of urate crystal.
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10.2Alkaline Urine

The pH scale ranges from 0 to 14. A pH of 7 is neutral, whereas a pH

results below 7 is acidic and above 7 is alkaline. Urine has the
highest range of pH compared to other bodily fluids. If a person has
a high urine pH, meaning that it is more alkaline, it might signal a
medical condition such as kidney stones, (calculla), urinary tract
infection (UTI’s), kidney related diseases.
To help make your urine more acid you should avoid most fruits
(especially citrus fruits and juices). Milk and other dairy products
and other foods, which make the urine more alkaline. Eating more
protein and foods such as cranberries. (especially cranberry juice
with vitamin-C added), plums or prunes may also help.
 25

Crystal in Acidic urine:

Calcium ocalate: calcium oxalate stones are the most common type of
kidney syone. Kidney stones are solid masses that form in the kidney
.when there are high levels of calcium oxalate ,cystine or pgosphate and
too little liquid, there are diffrent type you have.Calcium oxalate stones
are caused by too much oxalate in the urine.
Oxalate is a natural substances found in many foods your body uses
food. For energy. Afrer your body uses what it needs, waste products
travel through the bloof stream to the kidneys and are remove through
urine.

Calcium ocalate
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Uric acid:Uric acid crystals can be diffrent types of shapes,

barrel,plate like diamond .they are typically orange.brown or yello in
colour .they can be found in normal urine when caused by a protein
rich -diet which increases uric acid in the urine .uric acid kidney
stones often red orange brown in colour,throught uric acid crystals
are colourless uric acid crystals are characterized by their needle
shape & strong double refraction in polarized light,where as crystal
of calcium pyrophosphate dihydrate which are found in
chodrocalcinosis have a more rhomboid appearance and limited
bire fringence (2,3)

Uric Acid Crystals.

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CRTYSTAL IN ALKALINE URINE

1.Triple phosphates Crystals:Triple phosphates crystals also known
as magnesium ammonium pgosphat3 crystals,are found in alkaline
urine.(pH greater than 7).The formation of magnesium ammonium
phosphates Crystals (triple phosphates crystals)is caused by a
combination of factors including decreased urine volume combined
with bacteria in the renal system that are capable of producing
ammonia and increasing the urine pH (such as proteus or klebsiella-
type bacteria).In individual with kidney stones the repeated
presence of triple phosphate could be predict the formation of a
struvite (magnesium ammonium phosphates)Stones .It should be
noted however ,that most patients with triple phosphates crystals
will never form stones.

Triple phosphates crystals.

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Amorphous urates: Amorphous urates (Na,K ,Mg,or ca salts) tend to

form in acidic urine and may have yellow or yellow brown color
rhomboid .Amorphous phosphates are similar in general appearance
,but tend to form in alkaline urine and lack colour and are coffin-lid
shape .The presence of urate crystals sometimes called amorphous
urate crystals is generally of little clinical significance crystals are
considered normal .It they are form solutes that are typically found in
the urine.
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Ammonium biurates:These crystals are brown sphere with spiky throns

.They almost resemble small bugs .They are often found in alkaline urine
,but they can also be seen in normal urine. Sometimes ammonium
biurates crystals only appear beacause the urine sample is old or has
been poorly preserved. The formation of ammonium urate crystals can
be caused by a combination of factors including decreased urine volume
or a condition that alkalinizes (Increase the pH of)urine,such as
vegetarian diet,urinary tract infection of some medications.In individuals
who have kidney stones,the repeated presence of amorphous urate
crystals in the urine may indicate the probable nature of stones.It should
be noted however,that most patients with amorphous urate crystals will
never form stone .
 30

Casts:Urinary casts are tiny tube- Shaped particle that can be found
whwn urine is examined under the microscope during a test called
urinalysis. Urinary casts may be made up of while blood cell,red blood
cells,kidney cells,or substances such as proteins or fat normally the
presence of casts in the urine is considered to be an unusual
finding.However small amounts,of hyaline casts ( between 0-2 casts per
low power field of the microscope).May be detected in the urine of
healthy individuals without necessarily indicating a serious condition
like kidney disease.
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Granular cast: Granular casts are a sign of many types of kidney disease.
Red blood cells casts mean there is a microscope amount of bleeding
from the kidney .They are seen in many kidney diseases.Renal tubular
epithelial cells casts reflect damage to tubule cells in the kidney.Casts in
urine from normal individuals are few or none( and are usually hyaline
or granular in nayure )Granular casts are generally the results of
degeneration of cells in cellular casts[13]Their significance lies with the
casts from which they were formed.Granular may also the results or
direct aggregation of serum proteins and other substances into a matrix
of Tamm Horsfall mucoprotein .Casts are clusters of urinary sediment
elements.(fat bodies)

Granular casts
 32

Hyaline casts: Hyaline casts are the only casts that should be observed in
normal urine. And not of particular clinical intrest .They can be
observation after intense exercise,In very concerted low- volume
urine,or during diuretic treatment. Hyaline casts are cylindrical and clear
with a low refractive index.So they can easily be missed on cursory
review under bright field microscopy,or in an aged sample where
dissolutions has occured whereas on the other hand, phase contrast
microscopy leads to easier identification. Hyaline casts are cellular casts
that consits of a protein matrix.Presence of an occasional hyaline cast
may be normal however ,the number of hyaline casts .Increase with
renal disease.

Hyaline casts