1 BACTERIAL CELL AND ITS ULTRASTRUCTURE - DRA.

MARY JOYCE SABORRIDO TEOXON Different Types of Microscope Light microscope uses beam of light Electron microscope uses beam of electrons - Both light and electron microscope are used to view bacteria - It depends only on the magnification TEM vs. SEM - TEM has a higher magnification than SEM used to determine different organelles e.g. golgi apparatus and mitochondria Confocal Scanning Laser Microscope uses laser Scanning probe microscope used to view atomic structure
Organism

depends on other organism for survival have DNA/ RNA eukaryotes

Prokaryotes Eukaryotes Neither both

prions Virus Fungi Parasite Bacteria *human cell is considered as eukaryotes Bacterial Morphology ---shape, arrangement--Spheres (single) Coccus (pairs) Diplococci Tetrad (cluster) Staphylococcus (chain) Streptococcus Rods Bacilli/ Bacillus single pair chain Spiral - Very thin structure - Can be seen using dark field microscope wherein the background appears dark while the organism is seen with light *Coccobacillus = coccus + bacillus *pleomorphic - capable of transformation of shapes - varied in forms and shapes Gram Stain Mostly used differential stain Can be gm (-)/(+) bacteria Discovered by Heinz Christian Gram Composition: VIAS crystal violet grams iodine Jcelimpin 06/07/2011

Important Microbial Group prions - smallest microbial organism - acellular - neither eukaryotes nor prokaryotes - contain mostly of CHON component viruses - contain either DNA or RNA - an obligate intracellular parasite - depends on the host for replication to occur - acellular bacteria - contain DNA or RNA - classified as gm (-) or gm (+) - prokaryotes fungi - eukaryotes - contain DNA or RNA parasites

2 alcohol/ acetone safranin *In differential stain, the primary stain is the CRYSTAL VIOLET and the counter stain used is the SAFRANIN. Grams iodine - Enhanced the affinity of the crystal violet to bacterial cell - Its a mordant (not a stain) - Yellowish brown in color (amber) Alcohol/ acetone - Remove the primary stain - Serves as decolorizer 2 Types of Bacteria Gm (-) - With lipopolysaccharides - Able to washed away the crystal violet and iodine complex - Able to dissolve fats Gm (+) - No lipid component - Not able to remove the crystal violet iodine complex Components Gm (+) Gm (-) Crystal violet Grams iodine Alcohol/ acetone Safranin Violet Violet Violet violet
Ribosomes

Mycobacterium Corynebacterium Bacillus Clostridium Actinomyces Streptomyces Less common bacilli: Erysipelothrix Listeria Nocardia Bacterium ( Bifido-, Propioni-, Eu-. Arcano-) Gram Reaction Streptococcus gm (+) Staphylococcus gm (+) Neisseria gm (-) Bacillus gm (+) Escherichia coli gm (-) Structures found in gm (-)/(+) bacteria
Gm (-) Outer membrane Lipoprotein Cytoplasm Inclusion body Nucleoid body Both Gm (+) Septum

Mesosome

Violet
Inner membrane

Colorless

Cell wall Capsule (outermost layer)

Violet

Red/ pink
pili Flagella Porin CHON

flagella

Rules in Gram Staining All cocci are gm (+) except: Neisseria Moraxella (previously branhamella) Veillonella All bacilli are gm (-) except:

---cell envelope---

Techoic acids Lipoteichoic

LPS

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acids Phospholipids Outer membrane CHON Binding CHON lipoprotein Periplasmic space Thin cell wall Petidoglycan Phospholipids Inner membrane CHON cytoplasm Outer membrane major permeability barrier essential component of gm (-) Gm (-) Phospholipid/ CHON lipopolysaccharides Lipid A polysaccharides Hydrophobic membrane LPS = endotoxin Lipid A = toxic moiety PS = immunogenic Thick cell wall

K illers Klebsiella pneumoniae H ave Haemophilus influenzae N ice Neisseria meningitidis C apsule Cryptococcus neoformans (only fungi that is capsulated that cause meningitis in an immunocompromised individual BESS Bacillus anthracis poly-D glutamates capsule Escherichia coli meningeal stains have capsule mostly K1 Salmonella typhi (virulence) capsular antigen Streptococcus pyrogenes when 1st isolated, non immunogenic (but antiphagocytic hyaluronic acid capsule)
Gm (+) or Gm (-) Chemical composition Function

Envelope structure

Bacterial Envelope
Envelope structure Gm () or (+)
Both gm (-) and (+)

Chemical composition

Function

capsule -slime -glycocalyx

Composed of polysaccharide gel except Bacillus anthracis w/c is made up of polyD glutamic acid/ glutamate

pathogenecity -virulence factor -protect against phagocytosis e.g. macrophage, neutrophils and PMNs until opsonized immunogenic

*O-antigen = cell envelope antigen of a bacteria Structur Gm (-) or Chemical Functio e Gm (+) Composition n
Cell wall = peptidoglycan Either gm (+), thick 85% or gm (-), thin 15% found in all living bacteria except Mycoplasm Peptideglycan , polymer of peptides (typically 4 a.a. Cross linked to other chains) and glycans (made up of alternating Rigid support, cell shape, protecttion from osmotic damage, confer

S ome

Streptococcus pneumoniae

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a amino sugars) sugars found in peptidoglycan : Nacetylglucosamine (NAG) and Nacetylmuramic acid (NAM) e.g inhibit cell wall synthesis (penicillin) gm reaction, target of AB/ enzyme e.g. lysozymes

Structure

Gm (+) or Gm (-)
Gm (-)

Chemical Function composition

storage space between inner and outer membrane

Periplasmic space (non essential component)

*when inhibited cell become non viable Use for attachment of Gm (+) bacteria: Teichoic acid Lipoteichoic acid Structure Organism Chemical Function composition
Cell wall Acid fast only: this bacteria only resist acids specially in staining, once stained it is hard to decolorize Mycolic acid Acid fastness resistance to drying and chemical agents

Stores degradative enzymes w/c breakdown large molecules (Blactamases) acid regulation of osmolarity

Wall less forms of bacteria Spheroplasts (+) OM Protoplasts (-) OM Structure Gm ()/ (+) Chemical composition
Phospholipid bilayer w/ many embedded CHON, serves as the powerhouse of the bacteria

Function

Cytoplasmic / cell membrane (essential component)

Both gm -/+

All organism are non acid except: Cryptosporidium sp. Legionella sp Legionella micdodadei Isospora sp. Mycobacterium sp. Nocardia sp. (partially acid fast)

Energy metabolism occurs here, hydrophobic cell sack, selective permeability and active transport Adherence to cell surface (common pili either host or common bacterial cell)

Pilus/ fimbria 1.common 2.sex 3.virulence

Primarily gm (-)

Glycoprotein (pilin)

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including attachment to other bacteria during conjugation Flagellum/ axial filaments for Spirochetes; thicker than pili and with wavy structure Both spirochetes Gm(+) and (-); for motile bacteria CHON (flagellin) motility resistance, toxin prodn Ribosomes In eukaryotes, there is 80s target of some antibiotics e.g. erythromycin , chloramphenicol, clindamycin Both gm +/70s 30s 50s s=means Svedberg CHON synthesis

Types of flagella Monotrichous Lopotrichous tuft of flagella at one side Amphitrichous tuft of flagella on both sides Peritrichous Internal Bacterial Structure Gm +/- Chemical Function composition
Both gm + and Contain DNA, RNA, CHON Genetic material (all essential genes)

granules

Both gm +/-

Glycogen, lipids, polyphosphate, etc. Keratin coat, Ca++ dipicolinat e

storage

Endospores

Structure

Nucleoid region (essential component) also called nuclear/chromatin body/ nuclear region Plasmids (not essential components)

Gm (+) only e.g. Bacillus & Clostridium (major spore forming bacteria )

Resistance to heat, chemicals and dehydration *can survive in extreme temperature

*Bacillus subtilis common contaminants Bacteria w/ granules: Corynebacteria w/ metachromatic granules known as Babes-Ernst Granule Mycobacterium much granule Yersinia volutin granule

Both gm +/-

DNA

Non essential genetic material, role in conjugation , drug

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