BACTERIAL FUNCTION AND MORPHOLOGY FUNDAMENTAL SHAPES OF BACTERIA

I. ENVELOPE STRUCTURES
1. COCCUS
● round or spherical A. GLYCOCALYX - located outside the cell that contains polysaccharide
● morphologic variations: coffee bean shaped, lancet-shaped ● also called CAPSULE OR SLIME LAYER (called capsule when strongly adherent to the cell wall
● average diameter: 1 um and slime layer when loosely attached)
● examples: Staphylococcus, Gonococcus, Meningococcus ● function for adherence to cell surfaces
2. BACILLUS ● most bacteria potentially can have a capsule but is not essential to the growth of the bacteria
● rod-shaped but important in survival inside the host
● morphologic variations: club shaped, comma shaped, filamentous, etc. ● readily apparent as in S. pneumonia, B. anthracis or C. perfringens demonstrable by use of Hiss
● average size: 2 x 0.5 um capsule stain, appearing as clear space around the organism
● examples: Clostridium, Corynebacterium, Myobacterium ● size may be 2-3 X the diameter of the cell
3. SPIRILLUM ● Neufeld-Quellung reaction
● spiral or coiled organisms ○ capsular swelling reaction
● 3 forms: ○ addition of antiserum > capsule becomes swollen
○ Treponema - with fine regular coils ○ used for identification and typing of bacteria like S. pneumonia
○ Borrelia - coarse, irregular coils
○ Leptospira - very fine regular coils with hook at one or both ends ● significance of capsule:
● average size: may be as long as 40 um 1. antigenic - stimulates antibody formation
2. associated with virulence of the bacteria > delays ingestion of bacteria by phagocytes
(anti-phagocytic)
SIZE OF BACTERIA 3. transformable characteristic - genes for capsule formation can be transferred to other
● measured in terms of micrometer (1/1000 of a mm) or nanometer (1/1000 micrometer) bacteria
● size range of most pathogenic bacteria: 0.2-0.3 um up to 10 um ● composition of capsule:
○ among the smallest bacteria: Rickettsia, Mycoplasma ○ complex polysaccharide - Streptococcus, Enterobacter aerogenes
○ among the biggest bacteria: B. anthracis ○ polymers of D(-) glutamic acid - B. anthracis
● may be useful in the identification of bacteria ○ Hyaluronic acid - group A Streptococcus
● can be lost if grown repeatedly in culture

ARRANGEMENT OF BACTERIA B. CELL WALL (often referred to as murein sacculus) - composed of peptidoglycan (murein or
● arrangement is dependent on plane of division of bacteria and position taken after cell division has mucopeptide)
occurred ● functions:
● arrangement of cocci: 1. gives form and rigidity to bacteria
○ singly or in pairs (Pneumococcus, Gonococcus, Meningococcus) 2. provides protection to the bacterial cell
○ tetrads (Geffkya tetragena) 3. target of beta-lactam antibiotics
○ sarcinate (Sarcina lute) 4. lipoteichoic acid component - for attachment and adherence
○ clusters (Staphylococcus) 5. mycolic acid in acidfast bacteria - responsible for acidfastness
○ chains (Streptococcus) ● absent in Mycoplasma and Chlamydia
● arrangement of bacilli: ○ bacteria without cell wall - protoplasts
○ singly or in pairs (Klebsiella pneumonia) ○ bacteria with cell wall remnants - spheroplasts
○ chains (B. subtilis, B. anthracis) ● demonstrable by Dyar cell wall stain or by electron microscopy
○ palisade (M. leprae) ● principal structural component -> peptidoglycan
○ groups (M. tuberculosis) ○ multi-layered in gram positive bacteria and monolayer in gram negative bacteria
○ L, V, Chinese character arrangement (C. diphtheria) ● teichoic acid found in gram positive bacteria only
● arrangement of spirillum:
○ singly, pairs, groups. No typical arrangement found. Cell wall components:

A. PEPTIDOGLYCAN
STRUCTURE OF AN IDEALIZED BACTERIUM - 2% - 40% of cell wall dry weight
 - 2 amino acids: glucosamine and muramic acid 1. thin peptidoglycan external to the cytoplasmic membrane - 1-2% of cell wall dry weight
- chains may vary from less than 10 to more than 170 disaccharide units 2. outer membrane
- consists of polymeric backbones of repeating N-acetylglucosamine and N-acetylmuramic acid joined a. LPS - consists of lipid A which is the toxic moiety of LPS responsible for endotoxin
together by a tetrapeptide side chains and cross bridges activity
- diaminopimelic acid - an important element of bacterial cell wall which an immediate precursor b. porins - protein molecules that allow passive diffusion sugars and amino acids
of lysine (amino acid in position 3 of tetrapeptide chain), bacteria, it occupies position 3 c. ompA protein - anchor the outer membrane to the peptidoglycan layer and sex pilus
- stages of peptidoglycan synthesis: receptor for conjugation

1. Stages 1 & 2 FEATURES GRAM (+) GRAM (-)

- occur in the cytoplasm
- beginning with glucose, a series of reaction are carried out resulting in the formation of N Peptidoglycan Multilayered Monolayered
acetylglucosamine and attachment to a nucleotide uridine triphosphate (UTP) to form UDP
Complexity Simple Complex
derivative.
- subsequently synthesis results in the formation of UDP-N-muramic acid (M), plus a Teichoic acid Yes No
pentapeptide (AA)5
LPS No Yes

2. Stage 3 Lipoprotein & Phospholipid No Yes

- occurs in the cytoplasmic membrane
- UDP-M-AA5 is transferred to a lipid carrier, undecaprenol (UND), with release of UDP.
ACID FAST BACTERIA
- release of UDP results in peptidoglycan monomer UND-M-(AA)5 - migrate to the outer leaflet of
- Mycobacteria and Nocardia
the cytoplasmic membrane for peptidoglycan polymerization
- stain red with carbol fucshin and resist decolorization by acid alcohol
3. Stage 4
- MYCOLIC ACID - a substituted, B hydroxyl fatty acids that occur in Mycobacteria as esters bound to
- involves peptidoglycan cross-linkages
cell wall polysaccharides
- polymer migrates to nascent peptidoglycan to be incorporated in the cross-linkage
- contains equal amounts of peptidoglycan, arabinans and lipids
- more than 50% of lipid components are esterified mycolic acid
B. LYTIC ENZYMES
- 25% are normal fatty acids
- 3 major groups
- GLYCOLIPIDS *not cell wall bound
1. Endo-B-1,4 N-acetylhexosamidases
*trehalose, mycolates, sulfolipid, lipooligosaccharides, mycosides and
2. Endopeptidases
lipopolysaccharides
3. Amidases
Cell wall defective bacteria:
C. NON-PEPTIDOGLYCAN COMPONENTS
1. Protoplasts - with external surfaces free of cell wall constituents; osmotically fragile; usually derived
1. envelope proteins: e.g. M proteins
from gram (+) bacteria
2. capsular polysaccharides
2. Spheroplasts - have some remnants of the cell membrane, usually derived from gram (-) bacteria
- diverse capsule and surface polymers
3. L forms (L phase variants) - able to grow and divide and can revert to parent bacterial form
- soluble specific substance
- hyaluronic acid
C. OUTER MEMBRANE
- does not affect viability of the cell
- part of the cell wall in gram negative bacteria
- located external to the peptidoglycan layer
COMPARISON OF CELL ENVELOPE OF GRAM POSITIVE AND GRAM NEGATIVE:
- composition:
- lipid bilayer - attached to peptidoglycan by lipoproteins that cross the periplasmic space
Gram positive bacteria
- proteins - include porins which are involved in the transport of ions and hydrophilic compounds
- 30-200 molecules thick; 40-80% of total dry weight
- LPS - composed of lipid A, a polysaccharide-rich core, and a polysaccharide side chain
- strong rigid cell wall with several layers of peptidoglycan, teichoic acid and teichuronic acid
- polysaccharide: designated as 0 antigen
- usually devoid of proteins and lipids
- lipid portion: responsible for biologic effects of endotoxin
- functions:
Gram negative bacteria
1. maintains the structure of the bacteria
- more complex structurally and chemically than gram (+) bacteria
2. barrier to noxious environmental compounds
- 2 Layers
3. molecular sieve for small water soluble molecules
 FLAGELLA
D. PERIPLASMIC SPACE ● organ of locomotion; seen usually in bacilli
- fluid-filled space located between the outer membrane and the plasma membrane ● originates from cytoplasmic membrane
- contains hydrolytic enzymes and components of various transport systems ● slender whip like structures -> exhibit lashing, forward and rotatory movements
● made up of protein called flagellin
E. PLASMA MEMBRANE (Cytoplasmic Membrane) ● parts: basal body, hook, filament
- made up to 2 layers of lipids ● classification:
- the inside layer is consists of actin-like filaments which is responsible for the spiral appearance of ○ monotrichous - single polar flagellum
treponemes ○ lophotrichous - a tuft of flagella at one end or at both ends
- functions: ○ amphitrichous - single flagella at both ends peritrichous - flagella all around the bacillus
1. physical and metabolic barrier between interior and exterior of bacterial cell ○ atrichous - without flagella
2. Exhibits selective permeability ● demonstrable by electron microscopy or silver impregnation techniques
3. Location of bacterial electron transport system ● (Fisher and Conn flagellar or Leiffson stain)
4. Excretion of hydrolytic exoenzymes ● highly antigenic (H antigen)
5. Contains enzymes and receptors important in various systems of the cell ○ not vital to survival of bacteria
a. demonstrate by electron microscope ○ mutation or use of Waning blender can mean loss of flagella

II. INTERNAL or CYTOPLASMIC STRUCTURES PILI (FIMBRIAE)

● pili and fimbriae regarded synonymous (common pili)
A. BACTERIAL NUCLEUS (Nucleoid) ● thread-like or hair-like structures found in Gram negative bacteria
● not a true nucleus since it is not enclosed by a nuclear membrane and without nucleolus ● composed of structural proteins (pilins) & adhesins
● mitosis not demonstrable -> considered as prokaryotic cells ● fimbriae can be distinguished from flagella by its smaller diameter and it is not coiled
● appears as concentrated DNA in cytoplasm ● twitching motility
● consist of one double stranded, circular, covalently closed, supercoiled DNA molecule that is not ● demonstrable by electron microscopy
associated with histones ● types
● demonstrable by Feulgen staining or by electron microscopy 1. common pili - thousands of pili around bacteria
● bearer of hereditary characteristic 2. sex (F) pili - one or 2 in a bacterium
● functions:
B. MESOSOME 1. adhesion - adherence to glycoproteins of GUT; common pili
● formed by invagination of plasma membrane 2. used in transfer of genetic material by the process of conjugation (sex pili)
● lamellated or whorled-like structure 3. virulence
● associated with division septa during binary fission 4. antigenic - can induce antibody production
5. antiphagocytic
C. RIBOSOMES
● complex globular structures demonstrable by electron microscopy SPORES (Endospores)
● composed of RNA molecules and many associated proteins involved in protein synthesis ● resistant structures which enable bacteria to withstand adverse environmental conditions; convert to
● Size: 70S vegetative forms under favorable conditions
● composed of 2 subunits -> 50S and 305 ○ spores of C. tetani - can remain alive in soil for as long as 20 years and as long as 5 years in
● target of many antimicrobial agents like aminoglycosides chopped meat medium
● found only in few gram positive bacteria: Clostridium, Bacillus and a species of Rickettsia (Coxiella)
D. STORAGE GRANULES (Inclusion bodies) ● round, oval or elliptical structure; located terminally, sub-terminally or centrally in bacilli
● temporarily holds excess metabolites ● demonstrated by Dorner spore stain, Schaeffer & Fulton stain, heat and acetic acid method
● may be used for storage of food and energy, carbon, phosphates, etc. ● components:
● metachromatic granules of C. diphtheria made up of volutin or polyphosphates. 1. complete copy of chromosome
○ demonstrable by LAMB stain 2. minimum amount of proteins and ribosomes
● much granules of M. tuberculosis - gram positive granules 3. high amount of calcium bound to dipicolinic acid -> calcium dipicolinate
4. keratin-like protein coat
III. SURFACE APPENDAGES