PENJELASAN PRAKTIKUM

SISTEM UROGENITALIA

BAGIAN MIKROBIOLOGI
FAKULTAS KEDOKTERAN
UNIVERSITAS HASANUDDIN
2015

MICROBIOLOGY LAB
ASSISTANTS 2014-2015
MUH. RIDHO AKBAR
MUHAMMAD AZZAM
APRIZAL AZHAR
ANDI MUH OCTAVIAN PRATAMA
ANOM PARTHA JAYA
AGUNG DANAN JAYA
RADHI ARIFIN
MUH. FARID
FETTY ANDRIANI
FADHILAH PUTRI WULANDARI
NURUL FAJRINA
AISYAH NAULI SIHOTANG
MUFIDAH NUR QISTHI
SRI RINIA SARI ISWANTI
HANA AMIERA
AYU AQILAH BT. KHAZANI

TATA TERTIB
LABORATORIUM
MIKROBIOLOGI
Laki-laki :Memakai KEMEJA, CELANA PANJANG KAIN,
dan SEPATU TERTUTUP. Rambut TIDAK BOLEH
menyentuh kerah kemeja.
Perempuan : Memakai KEMEJA, ROK PANJANG KAIN,
dan SEPATU TERTUTUP. Rambut diikat ke atas dan bagi
yang berjilbab, jilbabnya dimasukkan ke dalam jas lab.

Perlengkapan yang WAJIB dibawa saat praktikum :

1.
2.
3.
4.
5.
6.
7.

Jas lab
Name tag
Penuntun praktikum + pas foto berwarna 4x6
Masker
Handscoen
Pensil warna
Penggaris 30 cm (per kelompok)

PERSYARATAN
ASISTENSI & RESPONSI

Telah mengumpulkan Tugas Pendahuluan

PRAKTIKUM
Telah mengikuti Asistensi & Responsi
Memenuhi Tata Tertib Laboratorium Mikrobiologi
Membawa Perlengkapan Praktikum
UJIAN PRAKTIKUM
Telah mengikuti Praktikum
Telah mengumpulkan Laporan Praktikum
Memenuhi Tata Tertib Laboratorium Mikrobiologi
REMEDIAL UJIAN PRAKTIKUM
Telah mengikuti Ujian Praktikum namun
dinyatakan lulus Ujian Praktikum

belum

INTRODUCTION

Urinary Tract Infection

Bacterial Agents
Causing UTI
Urine Sample &
Collection
Bacteriological
Examination

REVIEW :
BIOCHEMICA
L TEST

TSI (TRIPLE SUGAR IRON) TEST

Slant & Butt
Alkali : Merah
Asam : Kuning
Netral : Orange
H S
2
(+) : Hitam
(-) : Warna dasar
Gas
(+)
: Medium
terangkat/ pecah
(-) : Medium tidak
terangkat

SIM (SULFIDE INDOLE MOTILITY) TEST

H2S
(+) : Hitam
(-) : Warna dasar
Indol
(+)
: Terbentuk cincin
merah
(-)
: Tidak terbentuk
cincin merah
Motilitas
(+) : Stabline diffuse
(-)
: Stabline tidak
berubah

METHYL RED / VOGES PROSKAUER

(MR/VP)
Methyl Red

(+) : Merah
(-) : Kuning /
Orange

Voges Proskauer

(+) : Merah
(-) : Kuning /
Colorless

CITRATE UTILIZATION & UREA

HYDROLYSIS TEST
Citrate Utilization

(+) :
(-) :

Biru
Hijau

Urea Hydrolysis

(+) :

Pink

(-) :

Kuning

SUGAR FERMENTATION

Lactose Fermentation

Sucrose Fermentation

Glucose Fermentation

Mannitol

(+) :
(-) :

Kuning
Merah

URINARY TRACT

URINARY TRACT INFECTION

Aninfectionthat affects part of theurinary tract
caused

by

pathogenic

bacterial

infection

or

opportunistic infection.
Bacteriuria is a condition when the presence of
bacteria in urine with the number of 10 5 cfu in 1
ml of midstream urine or 103 cfu in 1 ml of urine
by supra-pubic aspiration.
Pyuria is the condition of urine containing pus in
urine, or white blood cell with the number of 5
cells/HPF or > 10 cells/mm3.

BACTERIAL AGENTS CAUSING

URINARY TRACT INFECTION

Escherichia coli (80%)

Coagulase-Negative Staphylococci (7%)

Staphylococcus saprophyticus in young woman

Proteus mirabilis (6%)

Other Gram-Negative Bacteria (4%)

Other Gram-Positive Bacteria (3%)

-Hemolytic Group B Streptococcus in pregnant

woman

BACTERIAL AGENTS CAUSING

URINARY TRACT INFECTION

Other organism

Anaerobic bacteria (2% rare)

Legionella pneumophila

Mycoplasma hominis

Ureaplasma urealyticum

Salmonella spp.

Shigella spp.

Adenovirus

Mycobacterium tuberculosis

Mycobacterium spp.

Campylobacter spp.
Chlamydia trachomatis
Gardnerella vaginalis
Haemophilus influenzae

INTRODUCTION
Escherichia coli
Klebsiella
pneumoniae
Enterobacter
agglomerans

Providencia
stuartii
Providencia
alcalifaciens
Alcaligenes
faecalis

Proteus vulgaris
Pseudomonas
aeruginosa

Proteus mirabilis

Escherichia coli
Family enterobacteriaceae
Gram-negative bacteria
Facultative anaerobic
Rod shaped
Commonly
found
in
lowerintestine
Have O, H, and K antigens

Esc
heri
chi
a

Slant
& Butt

Acid/Ac

H2S

Gas

Indole

Motili
ty

MR

the

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Klebsiella pneumoniae

Kle
bsi
ella
pne

Family enterobacteriaceae
Gram-negative bacteria
Facultative anaerobic
Rod shaped
Commonly found in the mouth,
skin, and intestine
Only have O and K antigens

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Proteus vulgaris

Pro
teu
s
vul

Family enterobacteriaceae
Gram-negative bacteria
Facultative anaerobic
Rod shaped
Inhabits the intestinal tracts of
humans, animals, and raw meat,
can also be found in soil, water,
and fecal matter

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Proteus mirabilis

Pro
teu
s
mir

Family enterobacteriaceae
Gram-negative bacteria
Facultative anaerobic
Rod shaped
Can
lead
to
the
formation
ofcrystalsofstruvite and calcium
carbonate
Part of the normal flora of human
gastro-intestinal tract, can also be
found free living in water and soil

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Enterobacter agglomerans

Ent
ero
bac
ter

Family enterobacteriaceae
Gram-negative bacteria
Also known as Pantoea agglomerans
Aerobic bacteria
Rod shaped
Inhabits plants (grows symbiotically
with various plants), soil, water, and
such
species
includes
bacteria
reported as both commensal and
pathogen of animals and humans

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Providencia alcalifaciens

Pro
vid
enc
ia

Family enterobacteriaceae
Gram-negative bacteria
Facultative anaerobic
Rod shaped
Could cause Travelers Diarrhea

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Providencia stuartii

Pro
vid
enc
ia

Family enterobacteriaceae
Gram-negative bacteria
Facultative anaerobic
Rod shaped
Commonly found in the intestine

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Alcaligenes faecalis

Alc
alig
ene
s

Family alcaligenaceae
Gram-negative bacteria
Obligat aerob
Rod shaped
Optimum growth occurs at about
37C

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

Pseudomonas aeruginosa

Family pseudomonadaceae
Gram-negative bacteria
Obligat aerob
Coccobacillus bacteria
Have O, H, and K antigens
Have 3 pigments :
Pyocyanin (blue - green)
Pyoverdine
fluorescent)

(yellow

green

and

Pyorubin (red - brown)

Pse
udo
mo
nas

Slant
& Butt

H2S

Gas

Indole

Motili
ty

MR

VP

Citrat
e

Sugar Fermentation
Urea

Gluco
se

Lacto
se

Sucro
se

Manni
tol

TSI

Acid

Alkali

Alkali

Acid

Acid

Neutral

Indole

H2S

MacConkey

+
Escherichia
coli

Klebsiella
pneumoniae

Proteus

Citrate
&
Urea

Klebsiella
pneumoniae

Brown

Brown + Green

Alcaligenes
faecalis

Pseudomonas
aeruginosa

Providencia

Indol

+
+

Escherichia
coli

Proteus
vulgaris

Proteus
mirabilis

Providencia
stuartii

Providencia
alkalifaciens

URINE SAMPLE & COLLECTION

The urine sample should be collected in a sterile manner
and put in a sterile container.
Sterilization cant be done with :
1. Wet heat sterilization

May decrease the volume of urine sample leading to

dilution of urine sample.
2. Chemical sterilization

May leave the chemical substance at the wall of

container and may kill the bacteria.
. Good urine for bacteriological examination is morning
urine, except for Mycobacterium tuberculosis
examination, 24 hours urine is usually used.

URINE COLLECTION

Midstream Urine

Urine Pocket
Supra-Pubic
Aspiration
Catheterization

MIDSTREAM URINE

Most commonly used for urine collection and bacteriological

examination of urine.

Urine specimen should be taken in the morning (morning

urine)

High risk of contamination (> 20%) and give a false-positive

culture.

Doesnt cause discomfort to the patient/client.

Easy to do and doesnt require any special skills and tools.

Difficult to do in patients with urinary incontinence.

Give a false-negative culture if use antiseptics to clean the

genital.

MIDSTREA
M
URINE

URINE POCKET

Usually used for uncooperative patients and collected 24

hours urine.

Medium risk of contamination (6-10%).

Easy to do and doesnt require any special skills.

URINE POCKET

SUPRA-PUBIC ASPIRATION

The most sterile method of collecting urine (specificity

99%)

The gold standard for bacteriological examination for

urine.

Requires to be done in the morning (morning urine) and

the bladder must be in full condition.

Requires any special skills and tools.

Cause discomfort to the patient/client (pain).

CATHETERIZATION

Should be avoided

Lead to irritation of urogenital tract mucosa and enhance

the infection

Low risk of contamination (95%)

Discomfort sensation for patients.

Must be done by a trained person.

COLLECTING, STORING,
TRANSPORTING, AND LABORATORIUM
Midstream Urine

Supra-Pubic Aspiration

Urine Pocket

Collecting

Sterile

Punction with spoit

Urine pocket or bag

Storing

Sterile bottle
at 4oc

Sterile bottle
at 4oc

Sterile bottle
at 4oc

Transporting

Laboratory

Ice box

Centrifuge
Gram staining

Isolation & Identification

Bacterial count

BACTERIAL EXAMINATION

.
.

Gram staining of direct preparation

Bacterial count
1. Pour plate method
2. Surface drop method
3. Dipslide method
Isolation and identification
Antibiotic sensitivity testing

GRAM STAINING OF DIRECT

PREPARATION

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)

POUR PLATE METHOD

POUR PLATE
METHOD

SURFAC
E
DROP
METHO
D

DIPSLIDE
METHOD

DIPSLIDE METHOD

ISOLATION & IDENTIFICATION

Urine is dripped onto the surface of MacConkey

/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)

PREDISPOSITION FACTORS UTI

Gender

Obstruction

Systemic infection

Pregnancy

Cateterization

Diabetes Mellitus

Tumor

Iritation

THA
NK
YOU