Faculty of Veterinary Medicine

AIRLANGGA UNIVERSITY

INTRODUCTION of
MICROBIOLOGY
I. THE HISTORY OF MICROBIOLOGY
MICROBIOLOGY
Consist of : science
- Virus
MICROORGANISM
- Bacteria associated with human
- Fungus ‘’JASAD RENIK’’ and animal life
- Algae
- Protozoa
-- first detailed BENEFICIAL HARMFUL
description

AS CAUSE OF
NORMAL FLORA DISEASES
Antony van Leewenhoek (1674)
The father of bacteriology MAKING WINE, CHEESE,
examined YOGHURT, BEER, ALCOHOL,
AND ANTIBIOTIC
RAIN WATER, SALIVA, TEETH

look at various forms

microorganisms
SPHERICAL, CYLINDRICAL , AND SPIRAL
MICROORGANISME FORMS are SPHERICAL (Coccus),
CYLINDRICAL (Bacillus), AND SPIRAL (Spirula)

HIS FINDING DOES NOT MAKE PEOPLE INTERESTED

people follow to

ABIOGENESIS THEORY

SPONTANEOUS
GENERATION
MICROORGANISM CAUSE THE DISEASE

PASTEUR ( 1877 )
succes
 ISOLATED BACILLUS anthaxis FROM THE BLOOD OF SICK ANIMAL
 FOUND FOWL CHOLERA DISEASES
ROBERT KOCH (1876) in Germany

 TALK ABOUT ANTHRAX BACILLUS

 IMPROVING THE TEHNIQUE IN MICROBIOLOGY :
STAINING METHOD
PURE CULTURE TEHNIQUE TO GET SPESIFIC MICROORGANISM

 KOCH POSTULATE DECLARATION :

1. A SPESIFIC MICROORGANISM CAUSE A SPECIFIC DISEASE
2. THE ORGANISM CAN BE ISOLATED AND GROWN IN PURE
CULTURE IN THE LABORATORY
3. THE PURE CULTURE WILL PRODUCE THE DISEASE WHEN IT’S
INOCULATED INTO A SUSCEPTIBLE ANIMAL
4. THE SAME MICROORGANISM MUST BE ISOLATED

 DISCOVERED TUBERCULOSIS AND CHOLERA DISEASES

 in 1900 ALL OF BACTERIA SPECIES TO CAUSE
VARIOUS OF DISEASES WAS KNOWN, LIKE :
CORYNEBACTERIUM DIPHTERIAE

SALMONELLA TYPHOSA

NEISSERIA GONNORHOEAE

CLOSTRIDIUM PERFRINGENS

CLOSTRIDIUM TETANI

SHIGELLA DYSENTRIAE

TREPONEMA PALLIDUM

ECT
 PREVENTION AND TREATMENT AGAINST THE DISEASES

IMMUNISATION ANTISEPTIC CHEMOTERAPEUTICA

& ANTIBIOTICA
EDWARD JENNER (1798)
PREVENT TO INFECTION
initially
often
USED COWPOX VIRUS TO previous
IMMUNIZE PEOPLE AGAINST SURGERY
SMALLPOX
JOSEPH LISTER
45% DIE used
using carbolic acid
PASTEUR (phenol)
- SURGICAL EQUIPMENT DIPPING
made - SURGICAL ROOM SPRAYING
VACCINE AGAINST : - COVERING OF THE SURGICAL INCISION
-FOWL CHOLERA DISEASE
- ANTHRAX CONTROL
SATISFYING RESULT
- RABIES
NOW
 OTHER CHEMICAL - ALCOHOL
- IODINE
 PHYSIS TECHNICAL
- AIR FILTER
- ULTRA VIOLET’S RAY
CHEMOTERAPEUTICA & ANTIBIOTICA
used
CONTROL OF DISEASES
OLDEST CHEMICAL AGENT :
- Hg2 Cl2 (AIR RAKSA ) SIPHILIS TERAPY
- QUININE/ KINA  AGAINST MALARIA

MODERN CHEMOTHERAPY
discoverer
-SALVARSAN  SIPHILIS PAUL EHRLICH
-SULFONAMIDA  INFECTION OF SEVERAL BACTERIA GERHARD DOMAGK

initially from
ANTIBIOTICA PENICILLIN FUNGI OF PENICILLIUM NOTATUM

INFECTION THERAPY WHICH NOT CURABLE BY SULFA

discoverer

ALEXANDER
FLEMING
II. CLASSIFICATION & NOMENCLATURE OF MICROORGANISM

CLASSIFICATION = TAXONOMIC SYSTEMATIC ARRANGEMENT OF

MICROORGANISM INTO GROUP
BACTERIA in a series

BERGEY’s MANUAL OF DETERMINATIVE BACTERIOLOGY (ed.8, 1974) • SPECIES

e.g.
• GENUS
KINGDOM : PROCARYOTAE • FAMILY
DIVISION : BACTERIA
CLASS : SCHIZOMYCETES – ending CETES
• ORDO
ORDO : EUBACTERIALES – ending ALES • CLASS
FAMILY : MICROCOCCACEAE – ending ACEAE • PHYLLUM/ DIVISION
GENUS : STAPHYLOCOCCUS • KINGDOM
SPECIES : AUREUS

based on
1. IDENTIFICATION
detect
CHARACTERISTIC OF MICROORGANISM

 MACROSKOPIC MORPHOLOGY OF COLONY

 MICROSCOPIC MORPHOLOGY OF BACTERIA CELL
 BIOCHEMICAL CHARACTERISTIC
 PHYSIOLOGICAL CHARACTERISTIC TO OXYGEN, pH & TEMPERATURE
 VIRULENCE/ PATHOGENICITY CHARACTERISTIC
 ANTIGEN STRUCTURE
2. COMPUTER = NUMERIC TAXONOMIC

COMPARED ONE DATA WITH OTHERS  LIST ARRANGE IN A SERIES BY

BASE ON COMMON CHARACTERISTIC e.g.:
 YES/ NO SPECIAL ENZYME
 YES/ NO PIGMENT
 YES / NO SPECIAL MORPHOLOGY STRUCTURE

3. GENETIC = GENETIC TAXONOMIC

DNA COMPOSITION ( GUANINE & CYTOSINE)

COMPARED WITH OTHER BACTERIA

% MOLECULE G + C CLOSE TO EQUAL

indicate
SAME CLUSTER
 BACTERIAL  PLANT NOMENCLATURE  THE BINOMIAL SYSTEM
NOMENCLATURE
LINNAEUS ( 1753 )

BINOMIAL NAME - CONSIST OF TWO LATIN WORDS

- THE FIRST WORD IS THE NAME OF THE GENUS
AND IS ALWAYS WRITTEN WITH A CAPITAL LETTER
- THE SECOND WORD IN THE NAME OF THE SPECIES
GIVING INFORMATION AND NOT CAPITALIZED AND IS USUALLY DESCRIPTIVE
ABOUT THESE BACTERIA
- ITALIC NAME ALWAYS, example : Staphylococcus albus

sometimes
SCIENTIFIC NAME have general name
example

 Brucella abortus
causing
abortus i.g.
Name of BRUCE

 Salmonella typhii
causing
Name of SALMON typhoid

 Neisseria gonorrhoeae gonococcus

 Mycobacterium tuberculosis Tubercle bacillus

III. METHODE OF MICROBIOLOGY

CLASSIFYING MICROORGANISMS

base on

IDENTIFICATION
observe characteristic of bacteria

MORPHOLOGY: FORM, SIZE,

STRUCTURE, STAINING REACTIONS
CULTURAL CHARACTERISTICS using
IDENTIFICATION
BIOCHEMICAL ACTIVITY
TECHNIQUE
ANTIGEN STRUCTURE/
1. MICROSCOPIC EXAMINATION
SEROLOGICAL CHARACTERISTIC
2. CULTURAL
PATHOGENICITY 3. BIOCHEMICAL ACTIVITY
4. SEROLOGICAL
* DNA (Genomic) 5. ANIMAL’S EXPERIMENTATION
6. PCR
* Protein profile
7. SDS-PAGE
 microscope BRIGHT FIELD (COMMONLY)
MICROSCOPIC EXAMINATION
DARK FIELD
determine ULTRA VIOLET
FLUORESCENCE
MORPHOLOGICAL CHARACTERISTICS PHASE CONTRAST
OF MICROORGANISMS/ BACTERIA ELECTRON
by
PREPARATION FOR LIGHT MICROSCOPIC EXAMINATION

A. NO STAINING BACTERIA IN LI- B. STAINING

( wet / native ) VING CONDITION

detect
CAPABLE of MOTION of BACTERIA
(motile / non motile)
SIMPLE NATIVE HANGING DROP
PREPARATION PREPARATION
 B. BY STAINING

NEGATIVE STAIN TECHNIQUE POSITIVE STAIN TECHNIQUE

INDIAN INK OR OBSERVE UNSTAIN BACTERIA

NIGROSIN  LEPTOSPIRA

SIMPLE
COMPLEX

THE BACTERIA APPEAR TRANSPARENT/ CLEAR ON A DARK BACKGROUND

 POSITIVE STAIN
TECHNIQUE

FIXED
SIMPLE STAINING PREPARATIONS COMPLEX

USE MORE THAN ONE

STAINING REAGENTS DIFFERENTIAL STAINING
DIFFERENTIAL STAINING

1. GRAM STAIN
reagents : CRYSTAL VIOLET
IODINE ( LUGOL ) SOLUTION
Gram +  violet ALCOHOL ACETON
Gram -  red SAFRANIN

2. ACID FAST (Ziehl Neelsen) STAIN for

Mycobacterium tuberculosis

reagents : CARBOL FUCHSIN HEATED have waxy substances

ACID ALCOHOL
METHYLENE BLUE
heating
- ACID FAST BACTERIA  RED
- NON ACID FAST BACTERIA  BLUE

SPECIAL STAIN

observe
SPORA (scaeffer fulton) STAIN LOCATION OF SPORA (CENTRAL,
SUB TERMINAL, TERMINAL
reagents : MALACHIT GREEN
SAFRANIN be needed
RED COLOR BACTERIA HEATING
GREEN COLOR SPORA
 Mikroskopis
 CULTURAL CULTURAL CHARACTERISTICS

FORM
by SIZE
COLOR
CONSISTENCY

PRIMARY ISOLATION  PURE CULTURE

SECONDARY ISOLATION  ISOLATE CULTURE
CULTURAL IDENTIFICATION

BIOCHEMICAL TEST
PRIMARY ISOLATION  STREAK PLATE TECHNIQUE

 PURE CULTURE

SECONDARY ISOLATION  ISOLATE CULTURE

MULTIPLY BACTERIAL
POPULATION

furthermore
IDENTIFICATION
Makroskopis
CULTURAL IDENTIFICATION

BIOCHEMICAL TEST AND SUGAR FERMENTATION

ANIMAL EXPERIMENT

BIOLOGICAL TEST PATHOGENICITY

propose

 DETERMINATION OF VIRULENCE/
PATHOGENICITY
 IDENTIFICATION  CLINICAL SIGN,
CHARACTERISTIC LESIONS
 PURIFY
 KOCH POSTULATE PROVED
HAPTEN/ANTIGEN TAK
SEMPURNA
*PROTEIN /PROTIN –IKATAN
*ASING BAGI INANG
*TAK MAMPU INDUCE ANTIBODY
*BEREAKSI DG ANTIBODI

ANTIGEN IMMUNOGEN
*PROTEIN /PROTIN –IKATAN *PROTEIN /PROTIN –IKATAN
*ASING BAGI INANG *ASING BAGI INANG
*INDUCE ANTIBODY *INDUCE ANTIBODY PROTEKTIF
*BEREAKSI DG ANTIBODI *BEREAKSI DG ANTIBODI

SEROLOGIS
*IKATAN ANTIGEN DG ANTIBODI
INVITRO
SEROLOGIS TES

AGLUTINASI
HA dan HI
PRESIPITASI
COMPLEMEN FIKSASI (CFT)
ENZIME LINK IMMUNO SORBENT
ASSAY(ELISA)
RADIO IMMUNO ASSAY (RIA)
FLOURESCENT ANTIBODIY TECHNIQUE
FLOWCYTOMETRY
SEROLOGICAL  RELATED WITH IMMUNITY

SEROLOGICAL TEST ANTIGEN + ANTIBODY REACTION

 REAKSI AGLUTINASI
TEMPAT PLATE
REAKSI
* PLATE
*MIKROPLATE
* TABUNG

+ -
AGID Test (AI)

1 2
Pour Agar Cut Agar

3 4 Fill Wells
Remove Agar Plugs
Hasil reaksi Agar gel
presipitasi
 Hemaglutinasi dan Hemaglutinasi
Neuraminidase

Hemagglutinin
Inhibition
M2

PB1
PB2
PA
HA
NP
NA
MA
HA untuk deteksi bakteri
yang mempunyai
NS

M1

Hemaglutinin

HI untuk menguji positif Contoh

penyakit yang disebabkan Coryza
oleh bakteri yang CRD
mempunyai hemaglutinin Cholera
TBC
ELISA (Enzyme-Linked
Immunosorbant
Assay)
ELISA Kit
Results
Instrument PCR

TBE Buffer
Genomik : Hasil PCR

1 2 3 4 5 M

Fig. 1. Agarose gel electrophoresis of ITS PCR products

Lanes 1–5, A. fumigatus, A. flavus, A. niger, A. nidulans and A. terreus, respectively.
M, 100 bp ladder molecular size marker (sizes in bp)
SDS-PAGE
 1. Protein preparation
 2. SDS PAGE preparation

stacking gel

resolving gel
Result protein/OMP bands on
SDS-PAGE
Western blot
Principle

 Transfer to nitrocellulose
membrane/PVDF

 Immuno-detection (indirect)
Instrument
Immunodetection Result
Thank you