UROGENITAL SYSTEM

BY;
NURHAFIZAH BT HATTA
C 111 09 865

Observe Gram Staining from Urine Sediment

Observe Biochemical Test

Urine Bacterial Count

Antibiotic Sensitivity Testing of Bacteria

Bacterial Agents of Urinary Tract Infections

Urogenital infections caused by pathogenic bacterial

infection
Bacteriuria the presence of bacteria 105 in the mid
stream urine
Pyuria the presence of white blood cell in the urine
Pyelonephritis UTI of the renal parenchym and/or
collecting system
Cystitis UTI associated with a superficial mucosal
invasion of the bladder

Some of the causative agents;

Escherichia coli

Staphylococcus saprophyticus

Streptococcus sp.

Proteus mirabilis

Other Gram negative bacteria

Must be collected in a sterile manner, to be put in a

sterile container.
Sterilization cannot be done with:
1. Wet heat sterilization may change the volume
of urine sample leading to dilution of urine
sample
2. Chemical sterilization will leave the chemical
substance at the wall of the container, thus
will kill the bacteria
Urine has to be collected in the morning

Mid stream urine

Urine pocket

Supra-pubic punction

Catheterization

Most commonly used urine sample in

bacteriological examination of urine
If properly done, may minimizing the
contamination or urethral normal flora

Area surrounding the external meatus is clean

with diluted detergent and clean water
(do not used disinfectants)
After removal the first excreted urine, next
urination is directly patched into a sterile
bottle (without touching the container or other
body parts of the patient)
Bottle is labeled and sent or kept in a cool place
(refrigerator, or with ice blocks)

Sterile bag plastic bag adhered around/in front

of external orificium to patch urine excreted
anytime by the patient

Sterile urine bag pocket available in market

Urine bag pocket is labeled and sent or kept in
a cool places (refrigerator, or in ice blocks)

The most sterile way of collecting urine.

(Gold standard ->99% specificty)

Requires to be done in the morning, as the bladder must

be in full condition

Supra-pubic is cleaned & sterilized with 70% alcohol

and bethadine or salvon.

By using 5ml sterile spoeit, puncture needle as deep as

finger on pubis until it penetrates the bladder.

Suck urine as many as 2-3ml, inject into a sterile bottle,

label and send to laboratory.

Urine catheterization should be avoided,

except for patients with permanent catheter.
Catheter lead to irritation of at urogenital
tract mucosa & enhance the infection
For patients with permanent catheter use
spoiet from the basal of disinfected catheter

Gram staining of direct preparation

Bacterial count
i) Pour plate method
ii) Surface drop method
iii) Dip slide method

Isolation & identification

Antibiotic sensitivity testing

Serotype / biotyping of bacterial agents

Direct preparation can be made from urine

sediments or urine without centrifugation.
From this preparation, we can see;
- The number of bacteria per field
- The number of leucocytes per field
(used as a criteria for pyuria)

a)

Pour plate method

b) Surface drop method

Urine is dipped with a sterile Pasteur pipette as

much as one to two droplets on the dry surface of
McConkey & Nutrient agar. Let the medium cool in
room temperature and incubate overnight at 350C
Bacterial growth:

Grade 0 no bacterial at the edge of droplet; 1 to 2

colonies only in the center of droplet
Grade 3 & 4 growth 105 cfu/ml; the edges of the
droplet will appear fully bacteria growth

c) Dip slide method

Sterile slide layered with agar medium at both

sides is dipped into urine(sample).
Dry the specimen & incubate overnight.

Bacterial growth then compared with standard

pattern.

Urine is dripped onto the surface of McConkey

/Nutrient agar/Blood agar/Chocolate agar.
Strike with a loop or a swab on the media surface
Culture is incubated overnight & the appearance of
growing colony observed.

*If there are 2 types of bacterial growth;

considered as contaminated (repeat)

Must be done
for observing
the bacterial
sensitivity to
antibiotics,
primarily to
those
commonly used
against
urogenital
infections.

For epidemiological purposes, sometimes it is

necessary to determine the serotype and
biotype of isolated bacterial agents.

Bacterial biotype can be also determined by

observing the pattern of antibiotic sensitivity
called biogram.

Triple sugar iron (TSI) reaction

- Observe slant, butt, H2S, Gas production
- Yellow (acid), Orange (neutral), Red (alkali)
- Black precipitate formed (H2S positive)

Sulphur Indole Motility (SIM) reaction

- Sulphur positive (black precipitate formed)
- Indole positive ( Red ring formed on the surface);
bacteria used trypthophanase enzyme to degrade
tryptophan to indole
- Motility positive ( Middle line production)

Methyl Red test ( Red = positive reaction)

Voges-proskauer test ( Red = positive reaction)

Citrate test ( Blue = positive reaction)

Urea test ( Pink = positive reaction) ;

bacteria used urease enzyme to degrade
urea to NH4
Sugar test ; Glucose, Lactose, Sucrose, Maltose
( Yellow = positive reaction)

Practical Work Guidelines, Urogenital System

2011
Medical Microbiology 24th Edition, McGraw Hill
(LANGE)