microbial mechanisms of pathogenicity · diseases caused by neurotoxins botulism •clostridium...
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CLINICAL MICROBIOLOGY
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The definition of clinical microbiology as a branch of
science dealing with the interrelation of macro- and
microorganisms under normal and pathological conditions
and in the dynamics of a pathological process with an
account of the treatment till the clinical and/or complete
recovery is presented.
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Other definition of Clinical Microbiology
The study of pathogenic microorganisms
such as bacteria, fungi, parasites and
viruses.
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Objectives Identify key clinical symptoms and the microbial immunological
conditions that characterize sepsis
Describe microbiology’s role to deliver sepsis-related
laboratory results and support Surviving Sepsis Campaigns
Describe Medical Laboratory Scientists’ (MLS) training
requirements for careers in medical microbiology laboratories
Define current standards and limitations of blood culture
methods and the premises for new diagnostic method
development to support rapid de-escalation of antibiotics
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Clinical microbiologists oversee
… - Operations to isolate and identify infectious
organisms (bacteria, viruses, parasites, or fungi)
from blood, urine, body fluid, sputum, wounds, or
other specimens
- Practices to provide crucial information that
guides the selection of antimicrobial therapy for
patients , as well as the proper specimen selection,
collection, and culture of pathogenic
microorganisms and normal flora.
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Services that are critical to the well-being
of sick patients by enabling the correct
diagnosis to be made
MLS Personnel, highly trained
professionals (board certified medical
laboratory scientists) who make countless
decisions each day that save lives an
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Clinical Microbiology Laboratories
• CML are the centerpiece of infectious disease diagnosis and
the cornerstone of infection control /prevention.
– Use laboratory techniques including culture, microscopic
examination, biochemical, molecular and immunological
tests.
–Identify the best course of antibiotic treatment by
establishing which antibiotics will be most effective in
combating the organism causing an infection.
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Clinical Microbiology Subspecialties
Bacteriology
Virology
Mycobacteriology
Mycology
Parasitology
Serology
Antimicrobial Testing
Molecular Microbiology
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Microbial Mechanisms
of Pathogenicity
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-Pathogenicity - ability of pathogento cause
disease by overcoming the defenses of the host
-Virulence - degree of pathogenicity
-Virulence factors – the various traits or
features that allow or enhance the microorganism’s ability to cause disease.
Many properties that determine a microbe’s
pathogenicity or virulence are unclear or
unknown
But, when a microbe overpowers the hosts
defenses, disease results!
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Portals of Entry
To cause disease, most pathogenic bacteria
must gain access to the host.
1. Mucus Membranes
2. Skin
3. Parenteral
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1. Mucus Membranes
A. Respiratory Tract
• microbes inhaled into
mouth or nose in
droplets of moisture or
dust particles
• Easiest and most
frequently traveled
portal of entry
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Common Diseases contracted via
the Respiratory Tract
Common cold
Flu
Tuberculosis
Whooping cough
Pneumonia
Measles
Strep Throat
Diphtheria
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Mucus Membranes
B. Gastrointestinal Tract
• microbes gain entrance thru
contaminated food & water
or fingers & hands
• most microbes that enter the
G.I. Tract are destroyed by
HCL & enzymes of stomach
or bile & enzymes of small
intestine
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Common diseases contracted via
the G.I. Tract Salmonellosis
• Salmonella sp.
Shigellosis
• Shigella sp.
Cholera
• Vibrio cholorea
Ulcers
• Helicobacter pylori
Botulism
• Clostridium botulinum
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Fecal - Oral Diseases
These pathogens enter the G.I. Tract at one
end and exit at the other end.
Spread by contaminated hands & fingers or
contaminated food & water
Poor personal hygiene.
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Mucus Membranes of the Genitourinary System - STD’s
Gonorrhea
Neisseria gonorrhoeae
Syphilis
Treponema pallidum
Chlamydia
Chlamydia trachomatis
HIV
Herpes Simplex II
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Mucus Membranes
D. Conjunctiva –
• mucus membranes that cover
the eyeball and lines the eyelid
Trachoma
• Chlamydia trachomatis
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2nd Portal of Entry: Skin
Skin - the largest organ of the body. When
unbroken is an effective barrier for most
microorganisms.
Some microbes can gain entrance thrugh
openings in the skin: hair follicles and sweat
glands
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3rd Portal of Entry: Parentarel
Microorganisms are deposited into the
tissues below the skin or mucus membranes
Punctures
injections
bites
scratches
surgery
splitting of skin due to swelling or dryness
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Preferred Portal of Entry
Just because a pathogen enters your body it
does not mean it’s going to cause disease.
pathogens - preferred portal of entry
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Preferred Portal of Entry
Streptococcus pneumoniae
• if inhaled can cause pneumonia
• if enters the G.I. Tract, no disease
Salmonella typhi
• if enters the G.I. Tract can cause Typhoid Fever
• if on skin, no disease
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Number of Invading Microbes
LD50 - Lethal Dose of a microbes toxin that will kill 50% of experimentally inoculated test animal
ID50 - infectious dose required to cause disease in 50% of inoculated test animals
• Example: ID50 for Vibrio cholerea 108 cells (100,000,000 cells)
• ID50 for Inhalation Anthrax - 5,000 to 10,000 spores ????
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How do Bacterial Pathogens
penetrate Host Defenses?
1. Adherence - almost all
pathogens have a means to
attach to host tissue
Binding Sites
adhesins
ligands(اتحاد)
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Adhesins and ligands are usually
on Fimbriae
Neisseria gonorrhoeae
ETEC
(Entertoxigenic E. coli)
Bordetella pertussis
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2. Capsules Prevent phagocytosis
attachment
Streptococcus
pneumoniae
Klebsiella pneumoniae
Haemophilus
influenzae
Bacillus anthracis
Streptococcus mutans
Yersinia pestis K. pneumoniae
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3. Enzymes
Many pathogens secrete enzymes that
contribute to their pathogenicity
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A. Leukocidins
Attack certain types of WBC’s
1. Kills WBC’s which prevents phagocytosis
2. Releases & ruptures lysosomes
• lysosomes - contain powerful hydrolytic
enzymes which then cause more tissue damage
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B. Hemolysins - cause the lysis of RBC’s
Streptococci
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1. Alpha Hemolytic Streptococci
- secrete hemolysins that cause the
incomplete lysis or RBC’s
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2. Beta Hemolytic Streptococci
- secrete hemolysins that cause the complete lysis
of RBC’s
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3. Gamma Hemolytic Streptococci - do
not secrete any hemolysins
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C. Coagulase - cause blood to
coagulate
Blood clots protect bacteria from
phagocytosis from WBC’s and other host
defenses
Staphylococci - are often coagulase positive
• boils
• abscesses
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D. Kinases - enzymes that dissolve
blood clots
1. Streptokinase - Streptococci
2. Staphylokinase - Staphylococci
Helps to spread bacteria - Bacteremia
Streptokinase - used to dissolve blood clots in the
Heart (Heart Attacks due to obstructed coronary blood vessels)
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E. Hyaluronidase
Breaks down Hyaluronic acid (found in
connective tissues)
“Spreading Factor”
mixed with a drug to help spread the drug
thrugh a body tissue
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F. Collagenase
Breaks down collagen (found in many connective
tissues)
Clostridium perfringens - Gas Gangrene
• uses this to spread thrugh muscle tissue
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G. Necrotizing Factor
- causes death (necrosis) to tissue cells
“Flesh Eating Bacteria”
Necrotizing fasciitis
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Summary of How Bacterial
Pathogens Penetrate Host Defenses
1. Adherence
2. Capsule
3. Enzymes
• A. leukocidins
• B. Hemolysins
• C. Coagulase
• D. Kinases
• E. Hyaluronidase
• F. Collagenase
• G. Necrotizing Factor
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4. Toxins
Poisonous substances produced by
microorganisms
toxins - primary factor - pathogenicity
220 known bacterial toxins
• 40% cause disease by damaging the Eukaryotic
cell membrane
Toxemia
• Toxins in the bloodstream
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2 Types of Toxins
1. Exotoxins
• secreted outside the bacterial cell
2. Endotoxins
• part of the outer cell wall of Gram (-) bacteria
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Exotoxins
Mostly seen in Gram (+) Bacteria
Most gene that code for exotoxins are
located on plasmids or phages
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3 Types of Exotoxins
1. Cytotoxins
• kill cells
2. Neurotoxins
• interfere with normal nerve impulses
3. Enterotoxins
• effect cells lining the G.I. Tract
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Response to Toxins
If exposed to exotoxins: antibodies against the
toxin (antitoxins)
Exotoxins inactivated ( heat, formalin or phenol)
no longer cause disease, but stimulate the
production of antitoxin
• altered exotoxins - Toxoids
Toxoids - injected to stimulate the production of
antitoxins and provide immunity
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Example: DPT Vaccine
D - Diphtheria
• Corynebacterium diphtheriae
P - Pertussis
• Bordetello pertussis
T - Tetanus
• Clostridium tetani
DPT - Diphtheria Toxoid
Pertussis Antigen
Tetanus Toxoid
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End of Lec-1
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Required Immunizations in Illinois 1. Diphtheria
2. Pertussis
3. Tetanus
4. Measles
5. Mumps
6. Rubella
• German Measles
7. Polio
8. Hib
9. Hepatitis B
10.Chicken Pox
Corynebacterium diphtheriae
Bordetello pertussis
Clostridium tetani
Measles virus
Mumps virus
Rubella virus
Polio virus
Haemophilus influenzae
Hepatitis B Virus
Varicella-zoster virus
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Type of Vaccines D
P
T
M
M
R
Polio
• Salk
• Sabin
Hib
HBV
Chicken Pox
Toxoid
Antigen
Toxoid
Attenuated
Attenuated
Attenuated
IPV – Inactivated Polio virus (Killed) 1953
OPV – Oral Polio vaccine (attenuated) 1964
Conjugated vaccine
Recombinant vaccine (antigen) yeast
• Capsid produced by genetically engineered yeast
Attenuated
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Most genes that code for exotoxins - plasmids
or phages
Lysogenic convergence
Diphtheria
Cytotoxin inhibits
protein synthesis -
resulting in cell death
Pseudomembrane
• fibrin, dead tissue,
bacterial cells
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Lysogenic Convergence
Scarlet Fever
Streptococcus pyogenes
• lysogenic convergence
prophage
• cytotoxin - damages blood capillaries and results in a skin rash
• Strep Thoat with a rash
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Diseases caused by Neurotoxins
Botulism
• Clostridium botulinum
• Gram (+), anaerobic, spore-forming rod, found in
soil
• works at the neuromuscular junction
• prevents impulse from nerve cell to muscle cell
• results in muscle paralysis
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Tetanus (Lock Jaw) Clostridium tetani
Gram (+), spore-forming, anaerobic rod
neurotoxin acts on nerves, resulting in the inhibition of muscle relaxation
tetanospasmin - “spasms” or “Lock Jaw”
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Diseases caused by Enterotoxins
Cholera
• Vibrio cholerae
• Gram (-) comma
shaped rods
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Cholera toxin
Converts ATP into cAMP
causes cells to excrete Cl- ions and inhibits
absorption of Na+ ions
Electrolyte imbalance
H2O leaves by osmosis
H2O Loss (Diarrhea)
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Severe cases, 12 - 20 liters of liquid lost
in a day
Untreated cases - Mortality Rate about 50%
Mortality may be reduced to about 1%
• administering fluids and electrolytes
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EHEC (Enterohemorrhagic E. coli)
E. coli (0157:H7)
enterotoxin causes a hemolytic
inflammation of the intestines
results in bloody diarrhea
• Toxin
• alters the 60S ribosomal subunit
• inhibits Protein Synthesis
• Results in cell death
• lining of intestine is “shed”
• Bloody Diarrhea (Dysentary)
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Endotoxins - part of the Gram (-)
Bacterial cell wall
LPS (Lipopolysaccharides)
• O Antigen
• Lipid A
Lipid A - Toxin portion of the LPS
• responsible for Fever that is associated with
many Gram (-) Bacterial infections
• Gram (-) cells are “digested” endotoxins are
released - fever
• Antibiotics