urogenitalia inter 2012

8/13/2019 Urogenitalia Inter 2012

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UROGENITALIAFatimah Yunikartika Akbar

Fakultas Kedokteran

Universitas Hasanuddin

2011

Penjelasan Praktikum Mikrobiologi


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ASISTEN MIKROBIOLOGI 2011 -2012

ASEPMETRIKA : Kel. A1-B1

MUH.AYYUB PRIMADI : Kel. A2-B2

MOH. DIMASISMAIL : Kel. A3-B3

MUH. YOGIPRATAMA : Kel. A4-B4

ANA ZAHARINA HASYIM : Kel. A5-B5

FATIMAH YUNIKARTIKA AKBAR : Kel. A6-B6

NUR HAFIZAH BT. HATTA : Kel. A7-B7 WINDAAGUSTIN : Kel. A8-B8

SUHAILAH BINTI MOHD JAMIL : Kel. A9-B9


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TATA-TERTIB LABORATORIUMMIKROBIOLOGI

Laki laki : Memakai celana panjang kain, no polo shirt, rambut

tidak boleh kena kerah baju

Perempuan :Memakai rok panjang kain, no polo shirt, rambut

diikat ke atas. Bagi yg memakai jilbab, kasi masuk di dalam jas lab

Alat yang harus dibawa saat praktikum :

Jas lab

ID Card

Penuntun praktikum + Foto 4x6 berwarna

Masker Handscoen

Penggaris panjang 1 / kelompok

Pensil Warna


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Urogenital infectionscaused bypathogenic bacterial infection

Bacteriuriathe presence of asmany as 105bacteria in one ml mid

stream urinePyuriathe presence of white blood

cell in the urine

URINARY TRACT INFECTIOS(UTI)


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AGENTS THAT CAUSE URINARYTRACT INFECTIONS

Escherichia coli: 80%

Coagulase-negative staphylococci: 7%

Staphylococcus saprophyticus in youngwoman

Other Gram-negative bacteria: 4%

Other Gram-positive bacteria: 3%

Streptococcus Group B in pregnant woman

Proteus mirabilis: 6%


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Other organism

Anaerobic bacteriarare(2%)

Mycobacterium tuberculosis Mycobacterium spp

Campylobacter spp

Chlamydia trachomatis

Gardnerella vaginalis

Haemophilus influenzae

Legionella pneumophila

Mycoplasma hominis

Ureaplasma urealyticum Salmonella spp

Shigella spp

Adenoviruses

URINARY TRACTINFECTIONS


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ESCHERICHIA COLI

- Enterobactericeae

- Negative Gram

- Rod Shaped- Non Spora

- Fakultative Anaerobic

- Indole (+)

- Citrate (-)

- Urease (-)

- H2S (-)- MR (+)

- VP (-)


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KLEBSIELLA PNEUMONIAE

- Enterobactericeae

- Negative Gram

- Non capsule

- Non motile- Facultative anaerobic

- Indole (-)

- Antigen O and K

- H2S (-)

- Citrate (+)

- Urease (+)

- MR (+)

- VP(-)

- Lactose fermentation


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ENTEROBACTERAGGLOMERANS

- Enterobactericeae

- Negative Gram

- Rod Shaped

- H2S (-)

-

VP (+)- MR (-)


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PROVIDENCIA

Providencia Stuartii

- Enterobactericeae

- Negative Gram

- H2S (-)

- Indole (+)

- Citrat (+)

- MR (+)- VP(-)

- Motile

ProvidenciaAlkalifaciens

- Enterobactericeae

- Negative Gram- Rod Shaped

- H2S (-)

- Indole (-)

- MR (+)- VP (-)

- Motile


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PROTEUS

Proteus Vulgaris

- Enterobactericeae

- Negative Gram

- Rod Shaped- Motile

- H2S (+)

- Indole (+)

- MR (+)

- VP (-)

Proteus Mirabilis

- Enterobactericeae

- Negative Gram

- Rod Shaped- Fakultative Anaerobic

- Motile

- H2S (+)

- Indole (-)- MR (+)

- VP (-)


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ALKALIGENES FAECALIS

- Alcaligenaceae

- Negative Gram

- Rod Shaped

- Motile

- H2S (-)

- MR (-)

-

Citrate and Gas (+)- Oxidase (+)

- Catalase (+)


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PSEUDOMONASAERUGINOSA

- Pseudomonadaceae

- Negative Gram

- Rod Shaped

- Obligative Aerob

- Normal flora in human skin

- Have 3 kind pigmen :

- Pyocyanin (bluegreen)

- Pyoverdine (yellowgreenand fluorescence)

- Pyorubin (redbrown)


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URINE SAMPLE

Must be collected in a sterile manner, to beput in a sterile container.

Sterilization cannot be done with:

1. Wet heat sterilizationmay change thevolume of urine sample leading to dilution ofurine sample

2. Chemical sterilizationwill leave thechemical substance at the wall of thecontainer, thus will kill the bacteria


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URINE COLLECTION

Good urine for bacteriological examination ismorn ing ur ine, except for Mycobacteriumtuberculosis examination, 24 hoursurine isusually used.

Urine samples are collected in the followingways :

1. Mid Stream Urine

2. Urine Pocket3. Supra-Pubic Punction

4. Catheterization


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Most commonly used urine sample in

bacteriological examination of urine

If properly done, may minimizing the

contamination or urethral normal flora

MID STREAM URINE


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Area surrounding the external meatus is cleanwith diluted detergent and clean water (do notused disinfectants)

After removal the first excreted urine, nexturination is directly patched into a sterile bottle(without touching the container or other bodyparts of the patient)

Bottle is labeled and sent or kept in a cool place(refrigerator, or with ice blocks)

MID STREAM URINE


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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)

URINE POCKET


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URINE COLLECTION:

BAGGED OR CLEAN CATCHContamination rate of 10%

Incooperative patien>105CFUinfection likely

104105CFUsuspicious


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The most sterile way of collecting urine. (Goldstandard99% specificty)

Requires to be done in the morning, as thebladder must be in full condition

Supra-pubic is cleaned & sterilized with 70%alcohol and bethadine or salvon.

By using 5ml sterile spoeit, puncture needle asdeep 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.

SUPRA-PUBIC PUNCTION


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Urine catheterization should be avoided,

except for patients with permanent catheter.

Catheterlead to irritation of at urogenital

tract mucosa & enhance the infection

For patients with permanent catheteruse

spoiet from the basal of disinfected catheter

CATHETERIZATION


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URINE COLLECTION:TRANSURETHERAL CATHERIZATION

>105Colony Forming Units - 95%

specificity

104105CFUinfection is likely

103104CFUSuspicious


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Gram staining of direct preparation

Bacterial count

i) Pour plate method

ii) Surface drop method

iii) Dip slide method

Isolation & identification

Antibiotic sensitivity testingSerotype / biotyping of bacterial agents

BACTERIOLOGICALEXAMINATION


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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)

GRAM STAINING OFDIRECT PREPARATION


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a) Pour plate method

BACTERIAL COUNT


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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 incubateovernight at 350C

Bacterial growth:

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

colonies only in the center of droplet

Grade 3 & 4growth 105cfu/ml; the edges of the droplet

will appear fully bacteria growth

BACTERIAL COUNT


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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.

BACTERIAL COUNT


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Urine is dripped onto the surface of McConkey/Nutrient agar/Blood agar/Chocolate agar.

Strike with a loop or a swab on the media

surfaceCulture is incubated overnight & the appearance

of growing colony observed.

*If there are 2 types of bacterial growth;considered as contaminated (repeat)

ISOLATION &IDENTIFICATION


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Must be done forobserving thebacterialsensitivity toantibiotics,primarily to those

commonly usedagainst urogenitalinfections.

ANTIBIOTIC SENSITIVITYTEST


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For epidemiological purposes,

sometimes it is necessary to determine

the serotype and biotype of isolatedbacterial agents.

Bacterial biotype can be also determinedby observing the pattern of antibiotic

sensitivity called biogram.

SEROTYPE & BIOTYPE ofBACTERIAL AGENTS


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BIOCHEMICAL

TEST


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TSI (TRIPLE SUGAR IRON)TEST

Slunt & Butt

Alcali : Red

Acid : Yellow

Neutral : Orange

H2S : (+) : Black

(-) : Basic Colour

Gas : (+) : Medium terangkat(-) : Medium tidak

terangkt


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SIM (SULPHUR INDOLEMOTILITY) TEST

Sulphur (H2S) : (+) : Hitam

(-) : Tidak ada

Indole :

Reagen kovacsberekasi dengan

indole dan menghasilkan warnamerah di bagian atas dari tabung.

bacteria used trypthophanaseenzyme to degrade tryptophan toindole

(+) : Terdapat cincin merah

(-) : Tidak ada cincin merah

Motility :

(+) : Bergerak (Stab line)

(-) : Tidak bergerak


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METHYL RED /VOGES PROSKAUER (MR/VP)

Methyl Red

(+) : Red (-) : Yellow

Voges Proskauer

(+) : Red (-) : Yellow


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CITRATE & UREAHYDROLYSIS

Citrate

(+) : Blue (-) : Green

Urea Hydrolysis

(+) :Pink (-) : Yellowbacteria used urease enzymeto

degrade urea to NH4


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SUGAR FERMENTATIONTEST

Lactose Fermentation

SucroseFermentation

GlucoseFermentation

Mannitol

(+) : Yellow (-) : Red


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RESPON!!!


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WRITE DOWN

YOUR NAME, ID,GROUP,

ASSISTANTSNAME, and CODEA


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1

Write down 5 agents

that cause UrinaryTract Infections!


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2

Explain about

bacteriuria!


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3

What is the characteristic of :

a. Escherichia colib. Providencia stuartii

Minimal 5


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4

Write down 3 methods of

bacterial count!


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5

Good urine for bacteriological

examination is (..a..), except

for (..b..) examination, (..c..) is

usually used.


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6

Write down predisposition

factors of Urinary Tract

Infection (UTI) ! (min.5)


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7

Write down 3 kinds pigmen of

Pseudomonas aeruginosa!


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8

Biochemical test :Alcali acid,

H2S (-), and Indole (-).

Bacteria?


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9

Write down the biochemical test and

the interpretation of :

a. b.


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10

Biochemical test :Alcali

neutral, Mc Conkey Brown.

Bacteria?


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RESPON!!!


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WRITE DOWN

YOUR NAME, ID,GROUP,

ASSISTANTSNAME, and CODE B


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1

Write down 5 agents

that cause UrinaryTract Infections!


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4

Write down 3 methods of

bacterial examination!


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5

a. Write down 4 methods of

collecting urine specimen!

b. What is the best method to

collect the urine specimen?


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6

Write down predisposition

factors of Urinary Tract

Infection (UTI) ! (min.5)


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7

Write down 3 kinds pigmen of

Pseudomonas aeruginosa!


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8

Biochemical test :Alcali acid,

H2S (+), and Indole (+).

Bacteria?


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9

Write down the biochemical test and

the interpretation of :

a. b.


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urogenitalia inter 2012

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