The Natural Ecosystem

Human Body
 Learning outcome
Students should be able to:
Describe the ecosystem of human body
Distinguish between normal flora and
pathogenic microbes
Explain the toxicology of pathogenic
microbes
Constant presence of m/os:
- through daily activities
- normal flora
- parasites
Parasites
- Little or no harmful effect
- Parasites causes damage pathogens
- Outcome of host-parasite relationship depends
on pathogenicity of the parasite
- Pathogenicity ability of parasite to inflict
damage on the host and on the resistance of the
host to the parasite
- Quantitative measure of pathogenicity = virulence
- Virulence expressed as cell or viral number that
will elicit a pathogenic response in a host within a
given time period
- Virulence and resistance of host are not constant
factors dynamic
- Measuring virulence

By experimental studies of LD50

More virulent ?? LD50
- Infection growth of microorganisms in the
host, whether or not the host is harmed
- Disease damage or injury to the host that
impairs host function
- Infection is NOT synonymous with disease.
- Normal flora microbial infections no
disease.
 Microbial Interactions with Higher Organisms

Favorable environments
Regions exposed to the environment
Not a uniform microbial environment
Each region differs chemically and physically
selective environment for certain m/os
Where infections (colonization and growth)
begin?
Mucous membranes
single or multiple layers of epithelial cells

direct contact with external environment
coated with protective layer of mucus
(glycoprotein) to protect against invasion
The invaders
Bacteria contact host tissues at mucous
membranes
attached loosely
attached firmly
Mucosal barrier breached invasion
Who has higher number and larger
variety of m/os?
SKIN vs. MUCOSAL MEMBRANES
Facts: Facts:
- visible exposed
- sheltered, moist
surface 2 m2
environment
(more stable)
- 400 m2
 skin
Normal flora of

Different region,
different chemical
composition and
moisture content
- Epidermis: periodic drying (not suitable)
- Glands: wonderful place

a. Eccrine glands
- sweat glands
- on palms, finger pads, soles of feet
- relatively devoid of m/os due to extensive flow
of fluid
b. Apocrine glands
- sweat glands
- underarm and genital regions (warm and humid)
- relatively high m/os

c. Sebaceous gland
associate with hair follicles
- secrete lubricant fluid (urea, amino acids, salts, lactic
acid, lipids)
- relatively devoid of m/os due to extensive flow of
fluid
Normal flora of skin
- Transient or resident
- Common residents (mostly g +ve)
E. g. Staphylococcus, Corynebacterium,
Propionibacterium acnes (G+ve)
Acinetobacter (G-ve)
Pityrosporum ovalis (yeast) on the scalp
Factors affecting normal flora of skin
a. Weather
b. Age
c. Personal hygiene

Limiting factors of growth of m/os on skin

a. Low moisture content
b. Low pH
Normal flora of
Oral Cavity
Includes teeth and tongue
Saliva - not a good microbial culture medium
- contains lysozyme and lactoperoxidase
Colonization begins
Fresh clean tooth surface
Formation of a thin organic film due to attachment of
acidic glycoproteins from the saliva
Attachment of bacteria (S. sanguis, S. sobrinus, S.
mutans and S. mitis).
Growth and formation of microcolonies formation
of plaque
Colonization of filamentous bacteria Fusobacterium
 Follow by spirochetes e.g. Borrelia sp., G+ve rods,
G-ve cocci and anaerobic filamentous Actinomycetes
Mouth O2. How to support anaerobic growth?

a. Presence of facultative anaerobes

b. Building up of plague dense matrix
Accumulation of dental plaque acid products
dental caries
 Two main types of m/os (dental caries)
a. Streptococcus sobrinus
- lactic acid-producing bacteria
- colonize smooth tooth surfaces
because of its specific affinity for salivary
glycoprotein
b. Streptococcus mutans
-lactic acid-producing bacteria
- found in crevices
-attach via dextran polysaccharide
only produce dextran in presence of sucrose
by means of dextransucrase
Streptococcus sobrinus Streptococcus mutans
 The role of oral flora in tooth decay is affected by

a. Tooth surface (frequent cleaning by tongue, cheek,

saliva, toothbrush), less crevices e.g. dogs tooth
b. Diet (high in sugar lactic acid decalcification
of enamel breakdown of tooth enamel
proteolysis of matrix of tooth enamel by proteolytic
enzymes released by bacteria)
Normal flora ofGI tract
Consist of stomach, small intestine and large
intestine
Stomach pH 2 (barrier)
Small intestine includes duodenum, jejunum
and ileum (pH 4-5)
Large intestine (pH 7)
increase number of m/os
The passage of food:

Water withdrawn concentrated feces

Bacteria one-third weight of fecal matter
Passage of material through gI tract = 24 h
Growth rate of bacteria is one to two doublings per
day
Normal flora of respiratory tract
Upper respiratory tract (nasopharynx, oral cavity and
throat) colonizes by m/os at the mucous membranes
but normally expelled by nasal secretions
Lower respiratory tract (trachea, bronchi, lungs) is
essentially sterile filtered by upper respiratory
tract and cilia.
Normal flora of urogenital tract
Bladder is sterile
Site of colonization epithelial cells lining of urethra
e.g. E. coli, Proteus mirabilis and etc.
Before puberty vagina is alkaline and does not
produce glycogen, L. acidophilus not found.
After puberty vagina is acidic, L. acidophilus
(ferments glycogen to lactic acid).
After menopause, glycogen disappears, pH rises and
flora resembles before puberty.
Pathogenesis of Pathogen
To alter host function and cause disease
Pathogenesis = ability of m/os to initiate
disease, includes entry, colonization, and
growth in host, resulting in changes in host
function and damage to host
1. Entry

- Must penetrate skin, mucosal

membranes or intestinal epithelium
- Wounds
2. Specific adherence

- Types of specificity

a. Tissue specificity: selectively adheres to cells

in particular region of body e.g. Neisseria
gonorrhoeae adheres to urogenital eplithelial
cells
b. Host specificity: selectively adheres to different
host e.g. humans, rodents or birds
- Substances that assist in adherence of bacteria
a. Polysaccharide synthesized and secreted by
bacteria e.g. capsule, glycocalyx or slime layer
adhesion to tissue and between bacteria
protection
b. Fimbriae and pili
E.g. E. coli mostly non-pathoghenic and are
part of normal flora. Few strains colonization
factor antigens (CFA) which are fimbrial proteins
colonize small intestine and initiate diarrhea.
 - M/os can cause harm through
a. Production of toxins
b. Invasion

3. Invasion (alternative)
- Can cause harm without invasion
- E.g. production of toxins, colonization after
antimicrobial chemotheraphy.
4. Colonization and Growth
- Multiplication of m/os

- Grow appropriate population disease

- Only grow with appropriate nutrients and

environmental conditions
- E.g. Brucellaabortus grows slowly in tissues of
infected cattle but rapidly in placenta (contain
erythritol) abortion
- Adaptation of m/os to nutrient poor environment
E.g. Iron deficiency will reduce growth of m/os.
Animals: iron bound to transferrin and lactoferin, thus
inavailable to m/os.
M/os: produce siderophore, an iron-chelating compound.
E.g. E.coli produce aerobactin (siderophore) removes
iron bound to transferrin.
- At the infection site, pathogen produces virulence
factors to establish and maintain disease.
- E.g. Streptococci, staphylococci, pneumococci, certain
clostridia produce hyaluronidase (break down
hyaluronic acid which act as tissue cement)
- Others e.g. proteases, nucleases and lipases
- Streptokinase (S. pyogenes) a fibrinolytic substance
dissolve fibrin clots
- Coagulase (S. aureus) cause fibrin clotting to protect
m/o from attack by host cells.
5. Host damage
Production of toxins to promote pathogenesis

Exotoxins
Enterotoxins
Endotoxins
 Exotoxins
Released extracellularly
Three categories

a) Cytolytic toxins
b) A-B toxins
c) Superantigen toxins
 Cytolytic toxins
Cause lysis of cells
Hemolysins lyse erythrocytes (blood agar plate)
Hemolysins also called lecithinases or
phopholipases
-toxin of C. perfringes, a lecithinase, dissolves
membrane lipids cell lysis
Streptolysin O (S. pyogenes) affects sterols of
host cytoplasmic membrane
Leukocidins (S.aureus) lyse white blood cells
decrease host resistance
Clostridium perfringens
 A-B toxins
Two covalently bonded subunits, A and B
B bind to cell surface receptor transfer of subunit A
across targeted cell membrane (functions to damage
cell)
Diptheria toxin by Corynebacteriumdiptheriae
Fragment B bind to host cell receptor proteolytic
cleavage of fragment A and B Fragment A enters
host cytoplasm disrupt protein synthesis by blocking
transfer of amino acid from tRNA to growing peptide
chain
Diphtheria skin lesion on the neck
 Superantigen toxins

Stimulate large numbers of immune response

cells inflammatory reactions
Toxic shock syndrome toxin and exfoliating
toxin A and B (Shigelladysenteriae and S.
aureus)
 Enterotoxin
Exotoxins that act on small intestine
massive secretion of fluid diarrhea
E.g. food poisoning m/os such as C.
perfringes, V.cholerae, E.coli
 Endotoxins
Cell bound and released in large amounts only
when cell lyse
E.g. lipopolysaccharide toxin of gram negative
bacteria
E.g. Escherichia,Shigella,Salmonella
Toxicity of endotoxins is lower than exotoxins.
E.g. LD50 of endotoxin is 200-400 ug/mouse but
LD50 of botulinum toxin is 25 pg/mouse (10
million times less)
Properties of exotoxins and endotoxins
Chemical properties
Exotoxins
- Proteins, excreted by certain g+ve or g-ve
bacteria, generally heat labile
Endotoxins

- Lipopolysaccharide-lipoprotein complexes,
released on cell lysis as part of outer
membrane of g-ve bacteria, extremely heat
stable
Properties of exotoxins and endotoxins Mode
of action/symptom

Exotoxins
- Specific, binds to specific cell receptors
or structures, defined specific action on
cells or tissues
Endotoxins
- General: fever, diarrhea, vomiting
Properties of exotoxins and
endotoxins Toxicity
Exotoxins
- Highly toxic, often fatal

Endotoxins
- Weakly toxic, rarely fatal
Properties of exotoxins and
endotoxins Immunogenicity
Exotoxins
- Highly immunogenic; stimulate

production of neutralizing antibody

Endotoxins
- Relatively poor immunogen; immune
response not sufficient to neutralize
toxin
Properties of exotoxins and
endotoxins Toxoid potential
Exotoxins
- Treatment of toxin with formaldehyde

will destroy toxicity, but treated toxin

(toxoid) remains immunogenic
Endotoxins
- None
Properties of exotoxins and
endotoxins Fever potential
Exotoxins
- Do not produce fever

Endotoxins
- Pyrogenic
 Comparison of Bacterial Exotoxins and Endotoxin
Exotoxin Endotoxin
Produced by both Gram-positive and Produced only by
Gram-negative bacteria Gram-negative bacteria
Released from cell Integral part of cell wall

Protein Lipid A of lipopolysaccharide

Very toxic Variable toxicity

Many types of exotoxin based
Only one type of endotoxin
on structure and function
Heat labile Heat stable
Specific receptors on Diverse range of host cells
host target cells and systems affected
Specific effects in host Diverse range of effects in host
Toxoidable :can be denatured to
Nontoxoidable: chemical composition
remove toxicity and retain
prohibits molecular modication
antigenicity
 Host Defenses

Different species, different susceptibilities to

same pathogen
Influenced by physiology, nutrition, anatomical
differences and tissue surface receptors.
Factor: age (young and old)
anatomical changes (prostate gland)
stress (cortisone)
diet (changes and malnourish)
 Host defenses
Nonspecific: against a variety of pathogens

Specific: directed against individual pathogens

Non-specific Host Defense System
Anatomical Defense
- Lysozyme in tears and other secretions
- Skin (physical barrier),
secretion of fatty acids and lactic acid

- Intestinal: rapid pH change

- Cilia and mucus
lining in respiratory tract
- Mucus lining in intestinal tract
- Flushing of
urinary tract
 Inflammation

Characteristics: redness, swelling, pain, heat

(localized)
Mediators of inflammation: cytokines which
are produced by leukocytes
Immediate localization of pathogen, often by
production of fibrin clot at site (crust)
 Fever
- Cause by pyrogenic m/os (g-ve)
- Slight fever: beneficial (accelerate phagocytic and
antibody responses)
- High fever (40oC): damage host tissues and cells
 Phagocytosis
- Occur when pathogens breaks through physical
and chemical barries (body secretions)
- Activate phagocyte which would lead to
a. Phagocytosis: engulfment and destroy of
pathogens by phagocytes (macrophages,
neutrophils and etc.)
b. Presentation of antigen on APCs induce
specific immunity
Specific Host Defense System
Presentation of antigen on APCs recognized by T
cells
Present on MHC class I (all nucleated cells)
recognize by Tc cells destroy cell directly
Present on MHC class II (macrophage, dendritic, B
cells) recognize by Th cells produce cytokines
activate other T cells (delayed type
hypersensitivity response) and B cells produce
antibody
 Germ-free animals
animals that have no microorganisms
living in or on it
raised within germ-free isolators in order
to control their exposure to viral,
bacterial or parasitic agents.
 Advantages:

a. Strictly calibrated
b. Will not spontaneously become sick due
to infections
c. Not able to infect humans and other
animals with undesirable m/os
 Disadvantages
Must be housed in total isolation
 Production of germ-free animals
The isolator
Separates an area from its surrounding
Sterilized with chemical disinfectants
Have working gloves, ventilating channels to conduct
sterile air in and out of isolator, entrance chamber (for
sterilization of objects intended to be introduced into
isolator)
- Feed, water, bedding, instruments needed are
packed into containers and sterilized in advance by
radioactivity, dry-sterilizing or autoclaving
 The animal
- Produced using hysterectomy: Remove entire
uterus aseptically, entrance chamber, uterus opened
and fetuses awakened.
- Produced caesarean section: Remove from uterus
and transfer isolator
- Can only avoid horizontal transfer but not vertical
transfer regular testing
 Characteristics of germ-free animals
(compare with conventional animals)

Caecum 5 times larger

Faeces is lighter in colour and odour-free
Do not degrade cholesterol to coprostanol or
bilirubin to urobilinogen (therefore colourless
and odourless faeces)
Reduced heart, lung and liver size
Size of lymphatic organs is reduced
Immunoglobulin concentration in serum is
reduced
 GNOTOBIOTIC ANIMALS

When known strains of bacteria or

microbiota are introduced to a germ-free
animal, it is renamed as a
gnotobiotic animal.
Laboratory animals affect research results
in unknown ways
Solution: to use microbiologically defined
animals
 laboratory animal raised under sterile
conditions
free of exposure to microorganisms
diet is controlled, preventing exposure to
microorganisms that may be in food
have lymphoid tissue that is not fully
developed and may have a deficiency of
serum immunoglobulin.
Specific pathogen-free animals (SPF)

laboratory animals that are guaranteed free of particular pathogens