UNIT 3.

MEDICALLY IMPORTANT MICROORGANISMS

A. VIRUSES
 Medically important microorganisms
 Viruses lack necessary cellular parts that can allow them to replicate
independent of the host cell.
 They also lack the genes and enzymes that are necessary for energy
production
 Obligate intracellular parasites
 Are acellular organisms
 Outer surface is called capsid (composed of capsomeres)
 Viruses possess only a single nucleic acid, either DNA or RNA, but never
both.

VIRUS CLASSIFICATION
 Presence or absence of an envelope
o Naked Viruses - whose capsids are not covered by an envelope
o Enveloped Viruses - covered with an envelope
BACTERIOPHAGES
 Are special type of viruses that primarily infect bacteria.
 They are similar to other viruses in that:
o They are obligate intracellular parasites
o Similarly shaped like other viruses
o Type of nuclei they possess
 They play a role in the acquisition of virulence factors of certain bacteria (e.g
diphtheria toxin of Corynebacterium diphtheriae) as well as in the transfer of
genetic material from one bacterium to another (transduction)

B. BACTERIA
 Medically important microorganisms
 Are prokaryotic cells with majority having an outer covering called the cell
wall (composed mainly of peptidoglycan)
 Unlike viruses they possess both the DNA and RNA
 Unlike eukaryotic organisms, bacteria possess a nucleoid instead of a true
nucleus, smaller ribosomes, and lack mitochondria

HANS CHRISTIAN GRAM

 Discovered Gram Stain in 1884 where pneumonia was a big problem
 Three causes of pneumonia; unknown (later identified as viral pneumonia, and
two types of bacterial pneumonia caused by either Streptococcus pneumoniae or
Klebsiella pneumoniae.

CAN BE CATEGORIZED INTO:

 Gram-negative bacteria with cell wall (e.g., E. coli)
 Gram-positive bacteria with cell wall (e.g., S. aureus)
 Acid-fast bacteria with lipid-rich cell wall (e.g., Mycobacterium tuberculosis)
 Bacteria without cell wall (e.g., Mycoplasma)

ILLUSTRATION OF GRAM-NEGATIVE AND GRAM-POSITIVE BACTERIA

C. FUNGI
 Are eukaryotic cells with an outer surface composed mainly of chitin.
 Their cell membrane is made up mostly of ergosterol
 Possesses both DNA and RNA
 They possess a true nucleus that is enclosed by a nuclear membrane and
mitochondria

Features Bacteria Fungi

Cell type Prokaryotic; unicellular Eukaryotic; unicellular or

multicellular

Role in ecosystem Can be both producers and Mainly decomposers

decomposers

Optimal pH Neutral pH (6.5 - 7.0) Slightly acidic (4.0 - 6.0)

Cell structures No true nucleus and Possess true nucleus and

membrane-bound organelles membrane-bound
organelles

Main component of Peptidoglycan, except in Chitin

cell wall archaebacteria

Sterols in cell Absent except in Present

membrane Mycoplasma

Mode of nutrition Heterotrophic, Heterotrophic; majority

chemoautotrophic, aerobic and facultative
photoautotrophic, aerobic, anaerobic
 anaerobic, facultative
anaerobic

Reproduction Binary fission Sexual and asexual

spores

D. ALGAE
 Are eukaryotic organisms whose outer surface consists primarily of
cellulose.
 They are plant-like organisms (most of them have chlorophyll and capable
of photosynthesis.
 Unlike plants, they do not possess true roots, stems and leaves
 Algae do not produce significant diseases in humans.
 Diatoms are unicellular algae that inhabit both fresh - and saltwater.
o Their cell wall contains silicone dioxide that may be utilized in
filtration systems, insulation and as abrasives.
 Dinoflagellates are also unicellular algae that are important members of
the phytoplankton group.
 Contribute greatly to the oxygen in the atmosphere and serve as
important links in the food chain
 Responsible also for “red tide”.
 These organisms produce powerful neurotoxin which are potentially
fatal disease called paralysis shellfish poisoning.

BACTERIAL MORPHOLOGY
MORPHOLOGY: BACTERIA
 Prokaryotic, have simpler structures compared to eukaryotic organisms.
 In terms of morphology, bacteria are classified into three basic shapes :
 Coccus (pl. cocci)
 Cocci can be described as spherical or round-shaped organisms
(e.g., Staphylococcus, Streptococcus)
 May be arranged singly, in pairs (diplococci), in chains
(streptococci), in clusters (staphylococci), in groups of four (tetrad),
or in groups of eight (octad)

 Bacillus (pl. bacilli)

 Are rod-shaped organisms (e.g., Escherichia coli, Salmonella)
 Some may be very short, resembling elongated cocci called
coccobacilli (e.g., Haemophilus influenzae).

 Spiral-shaped or curves
 Organisms may show variations in their morphology
 Vibrio cholerae (comma-shaped), Treponema pallidum (spiral-
shaped), and Corynebacterium diphtheria (club-shaped)
 ENVELOPE STRUCTURES
 Prokaryotic cells surrounded by a complex envelope that may vary in
composition. The envelope serves to protect the bacteria from harsh
environmental conditions.
 Glycocalyx
 Outermost covering of some bacteria.
 Gelatinous substances that are located external to the cell wall,
composed of polysaccharide or polypeptide, or both.
 Capsule: if strongly attached
 Slime layer: if loosely attached
 Capsule
 The possession of capsule makes some pathogenic bacteria resistant to
phagocytosis.
 The capsule is considered a virulence factor because it enhances the
ability of bacteria to cause disease.
 Capsule is found most commonly among Gram-negative bacteria, for
example, Escherichia coli, Neisseria meningitidis, Klebsiella pneumoniae,
Haemophilus influenza, and Pseudomonas aeruginosa.
 Some also found among gram-positive bacteria.

 Cell wall
 Sometimes called the murein sacculus.
 Its principal component is peptidoglycan, which is also called murein or
mucopeptide
 It is multi-layered in gram-positive bacteria and single-layered in
gram-negative bacteria.
 The cell wall provides rigid support and gives shape to the bacteria

SPECIAL COMPONENTS OF GRAM-POSITIVE CELL WALLS

1. Teichoic acids: comprise major surface antigens of gram-positive organisms and
can elicit antibody response.
 In some gram-positive organisms such as Staphylococcus aureus,
teichoic acids function for the attachment of the organism to the host cell.
 Provide tensile strength to gram-positive bacterial cell walls
2. Polysaccharides: polysaccharide molecules include neutral sugars such as
mannose, arabinose, rhamnose, and glucosamine.
 Also include some acidic sugars such as glucuronic and mannuronic acid.
SPECIAL COMPONENTS OF GRAM-NEGATIVE CELL WALLS
1. Outer membrane: a bi-layered structure where the inner leaflet is composed of a
lipopolysaccharide (LPS).
 It has special protein channels that allow passage of small or low-
molecular weight hydrophilic substances such as sugars and amino acids
 LPS has a complex glycolipid called lipid A, responsible for its
endotoxin activity. Located in the outer leaflet of the outer membrane.
2. Lipoprotein: functions to anchor the outer membrane to the peptidoglycan layer and
stabilizes the outer membrane.

3. Periplasmic space: a fluid-filled space between the outer membrane and the inner
plasma membrane. It contains enzymes for the breakdown of large transportable
molecules into transportable ones and enzymes that serve to detoxify and inactivate
antibiotics.

IMPACT OF GRAM STAINING OF THEIR CELL WALL

 Gram Positive Bacteria

o Primary stain: Crystal violet

o Mordant: Gram’s iodine (Fixes the crystal violet on the bacterial cell wall)

o Alcohol: Decolorizer (Retains the color violet due to its thick

peptidoglycan)

o Safranin: Will retain the primary stain (Violet)

o Result: Blue / Purple

 Gram Negative Bacteria

o Primary stain: Crystal violet

o Mordant: Gram’s iodine (Fixes the crystal violet on the bacterial cell wall)

o Alcohol: Decolorizer
(Removes the color violet)

o Safranin: Will lose the primary stain and will take the secondary stain
causing to appear red / pink

o Result: Red / Pink

FLAGELLA
 These are thread-like structures made up entirely of molecules of the protein
sub-unit flagellin.
 They project from the capsule and are organs for motility
 Classified into four types namely:
1. Monotrichous (single polar flagellum);
2. Lophotrichous (a tuft of flagella at one end of the bacterium)
3. Amphitrichous (flagella at both ends of the bacterium); and
4. Peritrichous (flagella all around the bacterium).
 Bacteria without flagella are called atrichous.
PILI OR FIMBRIAE
 These are rigid surface appendages found on many gram-negative bacteria.
 The are fine and short in comparison with flagella.
 Their structural protein subunits are called pilins.
 Pili may also function for motility.
 Function for adherence to cell surface (common pili) or attachment to another
bacterium during a form of bacterial gene exchange called conjugation (sex pili)

AXIAL FILAMENTS
 Axial filaments are also called endoflagella and are found in spirochetes (e.g.
Treponema pallidum causing syphilis and Borrelia burgdorferi causing Lyme
Disease)
 These are composed of bundles of fibrils, the structures of which are similar to
flagella.
 The filaments rotate producing movement of the outer sheath of the spirochetes
propelling them forward.

BACTERIAL GROWTH REQUIREMENTS

GROWTH
 As defined in medical dictionaries involves an orderly and organized increase in
the sum of all components of the organism
 Microbial growth is concerned with increase in the number of cells rather than
size
 Bacteria require nutrients and physical conditions that will promote their growth

NUTRITIONAL REQUIREMENTS
 CARBON
 Makes up the structural backbone or skeleton of all organic molecules.
 Based on their carbon source, microorganisms may be classified into
autotrophs (lithotrophs) and heterotrophs (organotrophs)
 Autotrophs are microorganisms that utilize inorganic compounds (e.g.,
carbon dioxide) and inorganic salts as their sole carbon source.
 Organotrophs are organisms that make use of organic substance like
sugars or glucose as their carbon source.
 For both, their energy may be derived from either light (photolithotrophs
and photoorganotrophs) or oxidation of inorganic substances
(chemolithotrophs and chemoorganotrophs).

 NITROGEN, SULFUR, PHOSPHORUS

 These are necessary for the synthesis of cellular materials like proteins
and nucleic acids.
 Nitrogen and sulfur are required for the synthesis of proteins
 Nitrogen and phosphorus are essential for the synthesis of nucleic acids
and ATP.

 INORGANIC IONS
 These include magnesium potassium, calcium, iron, and trace elements
(e.g., manganese, zinc, copper, cobalt).
 Magnesium: stabilizes ribosomes, cell membranes and nucleic
acids. Serves as a co-factor in the activity of many enzymes.
 Potassium is required for the normal functioning and integrity of
ribosomes and other enzymatic activities of the cell
o Calcium is important component of gram-positive bacterial cell wall and
contributes to the resistance of bacterial endospores against adverse
environmental conditions.
 o Iron is a component of cytochrome, a component of the electron transport
chain, and functions as a co-factor for enzymatic activities.
o Trace elements are components of enzymes and functions as co-factors.

 GROWTH FACTORS
 Growth factors are essential to promote the growth and development of
the bacterial cell.
 These include vitamin B and amino acids.

PHYSICAL REQUIREMENTS
 MOISTURE / WATER
 The bacterial cell is composed mainly of water. It serves as the medium
from which bacteria acquire their nutrients.

 OXYGEN
 Used by aerobic bacteria for cellular respiration and serve as the final
electron acceptor.
 Microbes are classified as either aerobes or anaerobes
based on their oxygen requirements.
 Aerobes: microorganisms that utilize molecular oxygen for
energy production
 Obligate Anaerobes: microbes that cannot survive in the
presence of oxygen
 Facultative organisms: organisms that can grow and survive
under both aerobic and anaerobic conditions.

 TEMPERATURE
 Enhanced enzyme activity requires certain temperatures.
 Categorized into three groups based on their temperature requirements:
(1) Thermophiles (which grow best at temperatures higher than 40
degree C;
(2) Mesophiles (require an optimal temp of 20 degree C - 40
degree C;
(3) Psychrophiles (require an optimum temperature of 10 degree
C - 20 degree C);
 Most medically important bacteria are mesophiles.

 pH
 Another requirement of bacteria is the extent of acidity or alkalinity of their
environment which is referred to as the pH.
 Microorganisms that grow best in pH 8.4 to 9.0 are called alkalophiles.
Those that grow best in pH 6.5 - 7.5 are called neutrophiles. Most
medically important bacteria are neutrophils.
 The pH of must human tissues are 7.0 to 7.2. Certain bacteria require a
pH less than 6.0. These bacteria are called acidophiles.

OSMOTIC CONDITIONS
 Most organisms grow best under ideal conditions of osmotic pressure, which is
determined by the salt concentration.
 The normal microbial cytoplasmic salt concentration is approximately 1%.
 The optimum condition is if the external environment also has the same salt
concentration.
 If the extracellular salt concentration is increased (e.g., when food is salted),
water will flow out of the microbial cell and the organisms will shrink and die.
  On the other hand, if the external environment does not contain salt, water will
flow into the bacterial cell causing the organism to swell and rupture.
 Organisms that require high salt concentration for growth are called halophiles
(e.g. diatoms and dinoflagellates) and those that require high osmotic pressure
for optimal growth are called osmophiles.

Bacterial Growth Curve

 Illustrates the phases in the growth of the population of bacteria when they are
grown in a culture of fixed volume.

STAGES OF GROWTH
 Lag phase
 Period of adjustment for the bacteria in the new environment.
 There is no appreciable increase in number of microorganisms.
 Microbes will show increased metabolic activity in order to synthesize
DNA as well as secrete enzymes which might not be present in their new
environment but which are needed by the organism.
 Bacteria attain their maximum size toward the end of the lag phase
 May last for 1 to 4 hours

 Log / Logarithmic / Exponential phase

 Characterized by rapid cell division; increase in number of bacteria
 Exhibits high metabolic activity
 Generation time or doubling time of the organisms (the time required the
bacterial cells to double in number) is determined.
 E.g., A generation time of 10 minutes means that the bacteria will double
in number every 10 minutes showing exponential growth.
 Average duration of this phase is about 8 hours

 Stationary Phase
 This is considered as the period of equilibrium.
 During this period, the rate of growth slows down, nutrients start to
deplete, and toxic wastes begin to accumulate - as a consequence some
to bacteria will die.
 The number of living cells equals the number of dead cells.

 Death or Decline Phase

 This is the period of rapid cell death where the number of dead cells is
greater than the number of living cells.
 The duration of this phase varies from a few hours to a few days.