Microbiology : Systematic study of Microorganisms

Microorganisms : Bacteria, Fungi, Algae,Parasites,Viruses,Prions and Viroids

Basic branches Bacteriology Study of Bacteria

Virology Study of Viruses Mycology Study of Fungi Parasitology Study of parasites Phycology Study of algae Prions and Viroids .

Applied Branches Medical Microbiology Study of microorganisms of medical importance

Agriculural Microbiology Study of microorganisms in Agriculture Food Microbiology Study of Microorganisms in Food Environmental Microbiology Study of Microorganisms in Environment Industrial Microbiology Study of Microorganisms applied in industries

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Medical Microbiology
Branches

Medical Bacteriology Study of Bacteria Medical Virology Study of Viruses Medical Mycology Study of Fungi Medical Parasitology Study of parasites Immunology Study of Immunity

Immunity : The resistance offered by host against

microorganisms or their parts or products

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Identification and Characterization of Microbes

BACTERIAL MORPHOLOGY

Cell shape or form is important for distinguishing different bacteria: Three Basic shapes: 1. Spherical 2. Rod 3. Spiral shape
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Identification and Characterization

Spherical shaped: Short rods: Rods: Curved rods: Spiral shaped: Branched bacteria: Irregular: Coccus Cocci Coccobacillus - Coccobacilli Bacillus Bacilli Vibrio vibrio Spirillum spirilla Actinomycete Actinomycetes Mycoplasma Mycoplasma

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Copyright 2004 Pearson Education, Inc., publishing as Benjamin Cummings

Different Arrangements of Bacteria

y y y y y y Cocci in Chains: Cocci in clusters: Cocci in pairs: Cocci in four: Bacilli in pair: Bacilli in chains: Streptococci Staphylococci Diplococci Tetrad Diplobacilli Streptobacilli

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Arrangements

Pairs: diplococci, diplobacilli Clusters: staphylococci Chains: streptococci, streptobacilli

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BACTERIA STRUCTURE
BACTERIAL CELL PARTS: Essential for Survival
Cell Wall Cell membrane Cytoplasm Nucleoids Mesosomes Ribosomes

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BACTERIA STRUCTURE
BACTERIAL CELL PARTS: Non-Essential for Survival
Flagellum Pili Capsule Endospore Intracellular Inclusions

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BACTERIAL CELL PARTS

1. Cell Wall
Cell wall: is the outermost component part.
The thickness of the cell wall ranges from 10-25 nm,

Cell wall is used: To Protects bacteria against lysis Allows them to grow over a wide range of osmotic pressures. It gives Rigidity to bacterial cells, which determines their characteristic shape.
 The main constituent of bacterial cell wall is a chemically complex polymer known as peptidoglycan.
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Cell Wall
Christian Gram recognized 2 different types of bacteria based on their staining reaction.

 Gram-positive bacteria stain purple and have thick layers of peptidoglycan combined with teichoic acid.  Gram-negative bacteria stain pink and have much thinner layer of peptidoglycan covered with an outer lipid membrane.

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Gram Stain smear on glass slide. 1. Prepare a thin bacterial Mechanism 2. Fix the smear by dry heat 3. Flood the smear with Crystal Violet for 1 min. 4. Wash the slide with tap water 5. Flood the smear with Gram s Iodine for 1 min. 6. Wash with tap water 7. Decolorize the smear by alcohol for 20-30 sec. 8. Wash slide with water 9. Flood the smear with Safranin 10.Wash with Tap water 11. Air dry slide and Observe under 100X oil immersion
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Gram negative Heat/Dry Crystal violet stain Iodine Fix Alcohol de-stain de stain Safranin stain
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Gram positive

Gram Stain Mechanism

Crystal violet -iodine crystals give blue coloration to cell Gram-positive
Alcohol dehydrates peptidoglycan CV-crystals do not leave

Gram-negative
Alcohol dissolves outer membrane and leaves holes in peptidoglycan CV- washes out

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Copyright 2004 Pearson Education, Inc., publishing as Benjamin Cummings

Figure 4.13b, c

The peptidoglycan is a complex, interwoven network that surrounds the entire cell and is composed of three parts: 1. Sugar (glycan) backbone which is composed of alternating N-acetylmuramic acid and N-acetylglucosamine 2. A set of identical tetrapeptide side chains attached to the N- acetylmuramic acid. 3. A set of identical peptide cross-bridges between two tetrapeptides.
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Peptidoglycan

Peptidoglycan
Polymer of disaccharide N-acetylglucosamine (NAG) & N-acetylmuramic acid (NAM) Linked by polypeptides

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Figure 4.13a

Gram-positive cell walls

Special Components of Gram-positive cell walls Contain significant amount of teichoic and

teichuronic acids, which account up to 50% of the dry weight of the cell wall.

 Teichoic acids are negatively charged fibers which protrude at the outer surface of Gram +ves.  In addition to this some gram-positive walls contain polysaccharide molecules.

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Gram-Positive cell walls

Teichoic acids: two types
Lipoteichoic acid links to plasma membrane Wall teichoic acid links to peptidoglycan

May regulate movement of cations

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Figure 4.13b

Gram-Negative Cell wall

Special components of Gram-Negative cell walls:  Gram-negative cell walls contain three components that lie outside of the peptidoglycan layer:
Lipopolysaccharide (LPS) Outer membrane Lipoprotein

 LPS - also known as endotoxin. The endotoxin is responsible for many of the features of disease, such as fever and shock (especially hypotension) caused by these organisms.
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Gram-Negative Cell wall

Periplasmic space = space between inner and outer membrane.
Gram-negative bacteria only have degradative enzymes located in this area.

Periplasmic enzymes act on the wide variety of substrates which a GN bacterium encounters in nature to convert them to molecules which are transportable into the cell.
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Gram-Negative Cell wall

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Figure 4.13c

Cell wall less forms in bacteria

 L forms : cell wall less forms of bacteria that appear spontaneously or due to treatment with Penicillium. Common in many bacteria E.coli, Neisseria.
Penicillin inhibits peptide bridges in peptidoglycan. Bacteria can revert back to cell wall forms.

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Cell wall less forms in bacteria

Lysozyme are enzymes that digest disaccharide in peptidoglycan.

 Protoplast : Spherical form of Gram Positive bacterium when treated with lysozyme.  Spheroplast : Spherical form of Gram Negative bacterium when treated with lysozyme.  Mycoplasma: Natural cell wall less forms and they have Sterols in plasma membrane
Protoplasts and spheroplasts are susceptible to osmotic lysis and are used for research.
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Cell wall of Acid-fast bacteria

 Mycobacteria have an unusual cell wall, resulting in their inability to be Gram-stained.
These bacteria are said to be acid-fast, They resist decolorization with acid-alcohol after being stained with Crystal violet.

 This is due to the high concentration of lipids, called as mycolic acids, in the cell wall.

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2. membrane also called cytoplasmic or Plasma Membrane The plasma

cell membrane The cytoplasmic membrane (5-10 nm thick) consists of proteins embedded in a bilayer of phospholipids Functions: Selective permeability, it controls which nutrients/ substances may transport into or out of the cell. Site for energy generation by oxidative phosphorylation Synthesis and assembly of cell wall components Section of enzymes and toxins Protects the cell against osmotic rupture
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Plasma Membrane

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Figure 4.14a

Plasma membrane Components of PlasmaMembrane Phospholipid bilayer Peripheral proteins Integral proteins Transmembrane proteins

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Figure 4.14b

Plasma Membrane
 Movement Across Membranes
Simple diffusion: Movement of a solute from an area of high concentration to an area of low concentration. Facilitated diffusion: Movement of solutes combined with a transporter protein in the membrane.

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Movement Across Membranes

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Figure 4.17

Movement Across Membranes

Osmosis
Movement of water across a selectively permeable membrane from an area of high water concentration to an area of lower water concentration.

Osmotic pressure
The pressure needed to stop the movement of water across the membrane.
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Figure 4.18a

Movement Across Membranes

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Figure 4.18c-e

Movement Across Membranes

Active transport of substances requires a transporter protein and ATP.
Substances such as amino acids, organic acids and inorganic ions combined with carrier proteins are transported across membrane and released inside the cell by this mechanism.

Group translocation of substances requires a transporter protein similar to active transport but the nutrients being transported undergoes chemical modification.
Sugars and fatty acids are transported by this mechanism.

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 The cytoplasm is the cellular material outside the nucleus but enclosed by the cell membrane.  Consists of 70% water  ~1000 different enzymes  Many ribosomes It is site of the bacterium for all activities It is controlled by the information carried in the DNA of the nucleus.

3. Cytoplasm

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Cell Wall

Cytoplasm

Cell membrane

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4. Nucleoid
Genetic material that controls all characteristics of cell . Centrally located cellular mass without a nuclear membrane. Contains a single circular chromosome, in which DNA is located.

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5. Mesosome and Ribosomes

Mesosome =It is an invagination of the cytoplasmic membrane, which forms septum during cell division.
Site for some respiratory enzymes. More seen in Bacilli.

Ribosomes =site for protein synthesis. Bacteria ribosomes are 70S in size with two subunits 50S and 30S.

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Ribosomes

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Figure 4.19

1. Flagella: are long (3-20 ), whiplike appendages that move the bacteria toward nutrients and other attractants, a process called chemo-taxis.
Made of chains of flagellin Attached to a protein hook Anchored to the wall and membrane

BACTERIAL CELL PARTS: Non Essential for Survival

The system enables bacteria to detect changes in concentration of certain chemicals and to move either toward (positive chemotaxis) or away (negative chemotaxis) from the substance depending on its nature.
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Copyright 2004 Pearson Education, Inc., publishing as Benjamin Cummings

Figure 4.8

Flagellar Arrangement in Bacteria:

 Monotrichous bacteria that have a single polar flagellum.  Amphitrichous bacteria having flagella at both ends.  Lophotrichous A tuft of flagella in a single location  Peritrichous bacteria flagella random distributed all over cell surface.
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Flagella Arrangement

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Figure 4.7

Motile Cells

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Figure 4.9

2. Fimbriae or pili
Pili are straight rigid rod like appendages. shorter and thinner than flagella, composed of a single protein pilin. Function :  Facilitate adherence of bacteria to other bacterial, red blood cells, or lining of the intestine.  Provide a site for attachment of bacteriophages (bacterial viruses).  Enable bacteria that have a sex pilus to transfer genetic material across a pilus bridge from one to another by a process known as Conjugation
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 Capsule plasma membrane secreted outside of the cell wall. It is polysaccharide that forms a protective outer coat to the cell. Can be detected by negative staining. Plays important role in bacterial virulence. Function: Protects bacteria from phagocytosis by white blood cells inside the body. Protects bacteria from external toxic substances & degradative enzymes. Provides water molecules to bacteria in dry conditions.
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3. Capsule produced by the is a thick gelatinous layer

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4. Endospores
Spore or endospore is a highly resistant, metabolically inactive forms of bacteria. Endospores are also formed during unfavorable temperature, pH and gaseous requirement. Endospore formation is protective measure and not reproductive process. The genetic material is enclosed in several protein coats that are resistant to heat, drying, freezing, toxic chemicals, and radiation. The process of Spore formation is - Sporulation The process of outgrowth or vegetative growth is Germination
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Copyright 2004 Pearson Education, Inc., publishing as Benjamin Cummings

Figure 4.21a

5.Intracytoplasmic inclusions:
Granules: are intracellular inclusions that serve as a storage for nutrients. Bacteria store food in reserve form during favorable conditions, when there is sufficient amount of nutrients & utilized during unfavorable conditions. Plasmids: are double stranded extrachromosomal DNA, particles that carry important genes for antibiotic resistance, toxin production, and carbohydrate fermentation. Plasmids number may vary from one to many.
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