Microbiology

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Microbiology Lecture Notes

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by www.easybiologyclass.com
(Sample but in‐depth and easy to follow lecture notes in microbiology for Students and Teachers of Biology/Life Sciences)

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Gram Positive Bacteria Vs Gram Negative Bacteria
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(Similarities and Difference between Gram Positive and Gram Negative Bacteria)
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What is Grams staining?
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Christian Gram, a Danish Physician in 1884 developed a staining technique to distinguish
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two types of bacteria. The two categories of bacteria based on gram staining are Gram
positive bacteria and Gram negative bacteria. Bacteria are first stained with crystal violet
or gentian violet. All bacterial cells will stain blue or purple colour with crystal violet
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solution. Then the bacterial cells are treated with iodine solution (Lugol’s iodine) solution
and washed with alcohol (de‐staining solution). Those bacteria which retain the blue or
purple colour of crystal violet are called Gram positive bacteria and those bacteria which
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loose the colour of crystal violet after washing with de‐staining solution is called Gram
Negative bacteria. Gram negative bacteria are later stained with safranin or fuchsin for
observation under microscope. Gram negative bacteria after safranin or fuchsin staining
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colour red or pink colour. Gram staining differentiates bacteria by the chemical and
physical properties of their cell walls by detecting the properties of peptidoglycan. Gram
staining method is useful in differentiating majority of bacterial species into two broad
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categories. Even though all bacterial species cannot be differentiated based on gram
staining technique, this method has immense application in clinical diagnostics and
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biological researches.
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Similarities between Gram Positive and Gram Negative Bacteria

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 Both are bacterial cells
 Both groups are prokaryotic
 Both lack membrane bounded organelles
 Both groups have covalently closed circular DNA as the genetic material
 Both groups contain extra‐chromosomal genetic materials (plasmids)

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 Both groups poses capsule
 In both groups, cell wall is made up of peptidoglycan

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 In both groups, cytoplasm is surrounded by lipid bilayer with many membrane
spanning proteins.
 Both gram‐positive and gram‐negative bacteria commonly have a surface layer

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called an S‐layer
 Both groups of bacteria undergo genetic recombination through transformation,

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transduction and conjugation
 Both groups undergo binary fission as a mode of asexual reproduction
 Both groups contain many flagellated and non‐flagellated species
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 Both gram positive and gram negative bacteria are inhibited by antibiotics (their
sensitivity varies)
 Both groups includes flagellated (motile) and non‐flagellated (non‐motile) forms
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Difference between Gram Positive Bacteria and Gram Negative Bacteria
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Gram Positive Bacteria Gram Negative bacteria
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Appears as dark violet or purple

Appears as pink or red coloured after
1 coloured under microscope after Gram
Gram staining
staining
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Retain the colour of crystal violet after Colour of crystal violet will not be
2 washing with alcohol (de‐staining retained after washing with de‐staining
solution) solution
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Examples: Bacillus subtilis,
Examples: Escherichia coli, Rhizobium,
3 Streptococcus, Lactobacillus,
Vibrio, Acetobacter

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Leuconostoc, Clostridium
Cell wall single layered, straight and Cell wall two layered and uneven
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even (wavy)

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5 Cell wall is very rigid and less elastic Cell wall is less rigid and more elastic

The rigidity of cell wall is due to the The elasticity of cell wall is due to the

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6 high proportion of peptidoglycans less amount of peptidoglycan (2 – 12
(80%) %)
Thickness of cell wall varies from 15‐ Thickness of cell wall varies from 7.5 to

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20 nm, sometimes up to 80 nm 12 nm

Muramic acid content of cell wall is Muramic content of cellwall is less, 2 –

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more, 16 – 20% of dry weight 5% of dry weight

Cell wall is resistant to alkalis and Cell wall is sensitive to alkalis and
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insoluble in 1% KOH solution soluble in 1% KOH solution

Cell is highly resistant of physical Cell is highly susceptible to physical

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disruptions disruptions
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S‐layer is attached to the peptidoglycan S‐layer is attached to the outer

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layer membrane
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12 Teichoic acid present in the cell wall Teichoic acid absent in the cell wall
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13 Periplasmic space absent Periplasmic space present

14 Outer membrane absent Outer membrane present

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15 Lipid content very low (1‐4%) Lipid content very high (11‐22%)
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Porins (proteinaceous membrane

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16 Porins are present

channels) are absent
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17 Lipopolysaccharides usually absent Lipopolysaccharides present

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18 Mesosomes are more prominent Mesosomes are less prominent

Produce endospores spores during

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19 Usually do not produce endospores
unfavourable conditions

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Flagella with four rings in the basal
20 Flagella with 2 rings in the basal body
body

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21 Usually produce exotoxins Usually produce endotoxins

Gram positive bacteria are more Gram negative bacteria are more

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susceptible to antibiotics resistant to antibiotics

Cannot be killed by vancomycin

23 Can be killed by vancomycin antibiotic
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antibiotic

Shows high resistance to sodium azide Shows low resistance to sodium azide
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solution solution
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Cells shows high susceptibility towards Cells shows low susceptibility towards
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penicillins and sulfonamide antibiotics penicillins and sulfonamide antibiotics
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Cells show low susceptibility towards Cells shows high susceptibility towards
26 streptomycin, chloramphenicol and streptomycin, chloramphenicol and
tetracyclines tetracyclines
Cell wall is highly susceptible to Cell wall is less susceptible to
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degradation by lysozyme enzyme degradation by lysozyme enzyme

28 Shows high tolerance towards dryness Shows low tolerance towards dryness
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29 Magnetosomes usually absent Magnetosomes sometimes present

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Very few are pathogenic to human and Most of them are pathogenic to human
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other animals and other animals

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Cell Wall Organization of Gram Positive and Gram Negative Bacteria

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Violet stained gram‐positive cocci and pink stained gram‐

negative rod‐shaped bacteria (Image source: Wikipedia)
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