Bakteri Gram Positif &

Bakteri Gram Negatif

Dicky B. Widhyatmoko
Dep. Mikrobiologi Kedokteran – FK UNAIR
September 27th 2016
Outline
• Pengertian Bakteri
• Pewarnaan Gram
• Klasifikasi Gram
• Bakteri Gram Positif
• Bakteri Gram Negatif
Pengertian Bakteri
• Definisi
▫ Single cell microorganism
▫ Ukuran 0.3 – 0.5 mm
▫ Memiliki tiga bentuk dasar : coccus, rods, curved /
spiral rods
• Perbedaan terletak pada cell envelope /
Identifying bacteria
• Size, shape, color
• Culturing techniques
• Metabolic attributes
• DNA
Staining / Pewarnaan
• Berfungsi untuk mewarnai bakteri
• Mudah untuk diamati / dikelompokkan
berdasarkan morfologi
• Pewarnaan dibagi menjadi 2 :
▫ Simple Staining  menggunakan satu zat warna,
ex. Methylene blue
▫ Differential Staining  menggunakan dua macam
zat warna
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The Gram Stain

• The Gram stain is named after the 19th century
Danish bacteriologist who developed it.
• Differential stain that distinguishes cells with a Gram-
positive cell wall from those with a Gram-negative cell
wall
▫ Gram-positive - retain crystal violet and stain purple
▫ Gram-negative - lose crystal violet and stain red from
safranin counterstain
• Important basis of bacterial classification and
identification
• Practical aid in diagnosing infection and guiding drug
treatment
Gram stain has four steps:

1. crystal violet, the primary stain

2. iodine, which acts as a mordant

by forming a crystal violet-iodine
complex

3. alcohol, which decolorizes

4. safranin, the counterstain.

G+ organisms are purple (or

bluish), G- organisms are red.
Cell Envelope
• Pelindung bagian luar dari cytoplasma
• Terdiri dari dua lapisan dasar :
▫ cell wall and cell membrane
• Menjaga keutuhan sel
• Perbedaan group ini berdasarkan pewarnaan gram
pada dinding sel dan sel membran
▫ Gram-positive bacteria: dinding sel terdiri dari dua
komponen utama yaitu peptidoglikan yang tebal dan
membran sel.
▫ Gram-negative bacteria: Memiliki membran sell
luar, lapisan peptidoglikan yang tipis, dan membran sel
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Gram-positive Cell Wall

• Thick, homogeneous sheath of peptidoglycan
▫ 20-80 nm thick
▫ includes teichoic acid and lipoteichoic acid:
function in cell wall maintenance and enlargement
during cell division; move cations across the cell
envelope; stimulate a specific immune response
Cell envelope of Gram positive
Is this gram stain positive or negative?
Identify the bacteria.
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Gram-negative Cell Wall

• Composed of an outer membrane and a thin
peptidoglycan layer
• Outer membrane is similar to cell membrane bilayer
structure
▫ outermost layer contains lipopolysaccharides and
lipoproteins (LPS)
 endotoxin that may become toxic when released during
infections
 may function as receptors and blocking immune response
 contains porin proteins in upper layer – regulate molecules
entering and leaving cell
▫ Bottom layer composed of phospholipids and lipoproteins
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Gram-negative Cell Wall

• Single, thin sheet of peptidoglycan
• Protective structure while providing some
flexibility and sensitivity to lysis
• Periplasmic space surrounds peptidoglycan
Cell envelope of Gram negative
Is this gram stain positive or negative?
Identify the bacteria.
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Classification Systems in the Procaryotae

1. Microscopic morphology
2. Macroscopic morphology – colony
appearance
3. Physiological / biochemical characteristics
4. Chemical analysis
5. Serological analysis
6. Genetic and molecular analysis
• G + C base composition
• DNA analysis using genetic probes
• Nucleic acid sequencing and rRNA analysis
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Species and Subspecies

• Species –a collection of bacterial cells which share
an overall similar pattern of traits in contrast to
other bacteria whose pattern differs significantly
• Strain or variety – a culture derived from a single
parent that differs in structure or metabolism from
other cultures of that species (biovars, morphovars)
• Type – a subspecies that can show differences in
antigenic makeup (serotype or serovar),
susceptibility to bacterial viruses (phage type) and
in pathogenicity (pathotype)
Common Pathogen
Gram Positive Bacteria
I-Gram Positive Cocci

A-Streptococcus (e.g. streptococcus Pneumoniae)

B-Staphylococcus (e.g. Staph. aureus)
C-Enterococcus (Previously Group D Strep.)

II-Gram Positive Rods

A-Corynebacteria: Corynebacterium diphtheria

B-Listeria monocytogenes
C-Bacillus anthracis (Anthrax)
D-Erysipelothrix rhusiopathiae

III-Gram Positive Branching Organisms

A-Actinomycetes
Gram Positive Cocci
• I-Beta-hemolytic Streptococcus (Lancefield Groups)
• - Group A Streptococcus (Streptococcus Pyogenes)
• - Group B Streptococcua (Streptococcus agalactiae)
• - Group C Streptococcus
• - Group G Streptococcus

• II-Alpha-hemolytic Streptococcus
• - Streptococcus Pneumoniae (Pneumococcus)
- Viridans streptococcus (bacterial endocarditis)

• III-Non-hemolytic Streptococcus
• - Streptococcus faecalis (Group D)
• - Certain members of Groups B, C, D, H, and O
Gram Negative
• Gram Negative Rods
• Anaerobes
▫ Bacteroidaceae (e.g. Bacteroides fragilis)
• Facultative Anaerobes (enteric/nonenteric)
▫ Enterobacteriaceae (e.g. Escherichia coli)
▫ Vibrionaceae (e.g. Vibrio Cholerae)
▫ Pasturella,Brucella,Yersinia

• Aerobes
▫ Pseudomonadaceae (e.g. Pseudomonas aeruginosa)
Facultative Anaerobes
• Enterobacteriaceae (e.g. E. coli)
• Vibrionaceae (e.g. Vibrio Cholerae)
• Salmonella,Shigella,Klebsiella,Proteus
• GI pathogens !!!!!
• non-enteric Pasturella,Brucella,Yersinia
• Francisella,Hemophilus,Bordetella