PHMP Midterms

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UNIT V: Microbial Control

Types of Microbial Control

1. Sterilization
- the removal or destruction of ALL living
microorganisms.

2. Commercial Sterilization
- Sufficient heat treatment to kill endospores of
Clostridium botulinum in canned food.

3. Sanitization
- Treatment is intended to lower microbial counts on FACTORS AFFECTING EFFECTIVENESS
eating and drinking utensils to safe public health
levels. 1. Microbial load
- associated with grains reflects the environmental
4. Disinfection conditions in which the crop was grown
- Destruction of vegetative pathogens on inanimate
objects. 2. Environmental influences
- (i.e., temperature)
5. Antisepsis
- Destruction of vegetative pathogens on living 3. Presence of organic matter
tissue. - (food residues) on the surface can interfere with
the antimicrobial activity of sanitizers and
disinfectants by reducing the level of activity or by
Other related terms protecting the microorganisms from attack by
acting as a physical barrier.
“Cide" - meaning to kill; treatment that causes death
of microorganisms. 4. Suspending medium
- Biocide or germicide - kills microorganisms in - the propagation of microorganisms or of living
tissue cells in special media conducive to their
general
growth
- Fungicide — kills fungi
5. Time of exposure
"-stat or -stasis" - meaning to stop or to steady; - Period in a sterilization process during which items
treatment that inhibits growth and multiplication of are exposed to the sterilant at the specified
microorganisms. sterilization parameters.
- Fungistatic - inhibits mold
- Bacteriostat -Inhibits multiplication of 6. Microbial characteristics
bacteria. - texture, transparency, and color or pigmentation.

Degerming - removal of microbes from a limited


area, such as the skin around an injection site. ACTIONS OF MICROBIAL CONTROL AGENTS

Sepsis - indicates bacterial contamination. 1. ALTERATION OF MEMBRANE


PERMEABILITY
Asepsis - absences of significant contamination. - The susceptibility of the plasma membrane is
due to its lipid and protein components.
Certain chemical control agents damage the
plasma membrane by altering its
RATE OF MICROBIAL DEATH
permeability.
When bacterial populations are heated or treated with
antimicrobial chemicals, they usually die at a constant 2. DAMAGE TO PROTEINS AND NUCLEIC
rate. ACIDS
- Some microbial control agents damage
cellular proteins by breaking hydrogen bonds
and covalent bonds. Other agents interfere
with DNA and RNA and protein synthesis.
fungi and its spores are killed within 10 minutes
Physical Methods of Microbial Control - EXCEPTIONS: X bacterial endospores

3. Pasteurization
- It uses a high temperature for a short time (720C
Dry Heat Sterilization for 15 seconds) to destroy the pathogens without
altering the flavor of the food.
- This is applicable for substances unaffected at a
temperature of 1480C to 2600C in the oven, at an
exposure time of 45 minutes.
- This method kills spores, as well as vegetative
forms of microorganisms.
- This method is ideal for sterilizing glasswares,
metalwares, and anhydrous oils.
- The principle of sterilization involved is the
oxidation of microorganisms by heat.

1. Direct flaming — used to sterilize inoculating


instruments using alcohol lamp or burner

2. Incineration — burning materials to ashes. 4. Fractional Sterilization Methods


- Are effective for vegetative forms of
microorganisms, but not for spores.

Methods of Fractional Sterilization

1. Tyndallization - It makes use of moist heat at 100


degrees Celsius, using free flowing steam.
- It is normally performed by 2 to 3 exposures,
alternated with intervals at room temperature or
incubator temperature.

2. Inspissation
- It is a fractional method of sterilization at 60
Moist Heat Sterilization
degrees Celsius in an oven, alternated with
intervals at room temperature or incubation for 2
- This is more effective than the dry heat method. to 3 days.
- The principle of sterilization is the coagulation of
the cell protein of the microorganism. 3. Filtration
- The passage of liquid or gas through a filter with
1. Autoclaving (steam under pressure) pores enough to retain microbes.
- It is the most effective method of moist heat - Microbes can be removed from air by high
sterilization. efficiency particulate air filters.
- It can destroy the spore formers. - Membrane filters composed of nitrocellulose or
- The temperature used is 121 C with a pressure of cellulose acetate are commonly used to filter out
15 psi for 15-30 minutes exposure. bacteria, viruses and even large proteins.
- Example: HEPA FILTER - can reduce the amount
of airborne allergens.

4. Cold
- The effectiveness of low temperatures depends on
the microorganisms and the intensity of the
application.
- Microorganisms do not reproduce at ordinary
refrigerator temperatures (00 to 70C)
- Many microbes survive (but do not grow) at the
subzero temperatures used to store food.

5. Desiccation
- In the absence of water, microorganisms cannot
grow but can remain viable.
2. Boiling
- Viruses and endospores can resist decolorization.
- Boiling point of water is 1000 C. At this
temperature, vegetative pathogens, viruses, some
6. Osmotic pressure
- Microorganisms in high concentrations of salts Evaluating a Disinfectant
and sugars undergo plasmolysis.
- Molds and yeasts are more capable of growing in In the use-dilution test, bacterial (S.choleraesuß, S.
materials with low moisture or high osmotic aureus, and P. aeruginosa) survival in the
pressure than bacteria are. manufacturer's recommended dilution of a disinfectant is
determined. In the filter paper method, a disk of filter paper
7. Radiation is soaked with a chemical and placed on an inoculated
- Use of short wavelength, high-intensity radiation to agar plate; a clear zone of inhibition indicates
destroy microorganisms. This radiation can come effectiveness.
in the form of gamma or X-rays that react with
DNA resulting in a damaged cell. Non-ionizing
radiation uses longer wavelengths and lower
energy.

Types of Radiation

1. Ultraviolet radiation - This is used to aid


reduction of air borne contamination produced by
mercury vapor lamps. This method has poor
penetration capability. Its effectiveness depends
on:
- Length of time of exposure Types of Disinfectants
- Intensity of radiation
- Susceptibility of the microorganism 1. Phenol and Phenolics
- Phenolics exert their action by injuring plasma
2. Ionization Radiation membranes, inactivating enzymes, and denaturing
- This radiation method makes use of high energy proteins.
emitted from radioactive isotopes such as cobalt
- Common phenolics are cresols and
60 (gamma rays) or by cathode or beta rays
(mechanical acceleration of electrons to high hexachlorophene.
velocity and energy).
2. Chlorhexidine
Note: Gamma rays are more reliable because there is no - Chlorhexihide damages plasma membranes of
mechanical breakdown, but it has a disadvantage of rare vegetative cells.
source and cannot be shut off immediately. - Chemical Methods of Microbial
Accelerated electrons provide higher and more uniform
dose outfit and can destroy organisms by stopping its
reproduction. Chemical agents are used on living tissue 3. Halogens
(as antiseptics) and on inanimate objects (as - Some halogens (iodine and chlorine) are used
disinfectants). Few chemical agents achieve sterility. alone or as components of inorganic or organic
solutions.

4. Alcohols
Chemical Methods of Microbial Control - Alcohols exert their action by denaturing proteins
and dissolving lipids.
- In tinctures, they enhance the effectiveness of
Principles of Effective Disinfection
other antimicrobial chemicals.
- Aqueous ethanol (60% to 90%) and isopropanol
The presence of organic matter, degree of contact with
microorganisms, and temperature should be considered. are used as disinfectants.

5. Heavy Metals and Their Compounds


- Silver, mercury, copper, and zinc are used as
germicidals.
- They exert their antimicrobial action through
oligodynamic action.

6. Surface-Active Agents
- Surface-active agents decrease the tension
between molecules that lie on the surface of a
liquid; soaps and detergents are examples. Two types of medication in selective toxicity:
- Soaps have limited germicidal action but assist in 1. Healthy Human Cells
the removal of microorganisms through scrubbing. 2. Mutated Cells
- Acid-anionic detergents are used to clean dairy
Chemotherapy: The use of drugs to treat a disease/
equipment. microorganisms
- Broad term for drugs that can kill a specific type of
7. Quaternary Ammonium Compounds cell
- Quats are cationic detergents attached to NH4+. - A form of antimicrobial drug
By disrupting plasma membranes, they allow Chemotherapeutic agents: Include chemicals that
cytoplasmic constituents to leak out of the cell. combat disease in the body.
They are most effective against gram-positive
Antineoplastic agents - Anticancer medication
bacteria. - Targets cancer cells without killing healthy cells

8. Organic Acids and Derivatives


- Sorbic acid, benzoic acid, and propionic acid Antimicrobial drug
inhibit fungal metabolism.
- They are used as food preservatives. - Is a chemical substance that destroys
pathogenic microorganisms with minimal
9. Aldehydes damage to the host tissues.
- Aldehydes such as formaldehyde and - Chemical that kills or inhibits the growth of
glutaraldehyde exert their antimicrobial effect by microorganisms.
inactivating proteins. - Broad Term
- They are among the most effective chemical
disinfectants. Antibacterial - Targets Bacteria
Antifungal - Targets Fungi
10. Gaseous Chemosterilizers Antiviral - Targets Viruses
- Ethylene oxide is the gas most frequently used for Antiprotozoal Targets infections caused by protozoa
sterilization.
Anthelmintic - Targets infections caused by
parasitic worms
- It penetrates most materials and kills all
microorganisms by protein denaturation.

11. Oxidizing Agents


- Ozone and peroxide are used as antimicrobial
agents.
- They exert their effect by oxidizing molecules
inside cells.

Antibiotic: Chemical produced by a microorganism


Note: Iodine is available as a tincture (in solution with
that kills or inhibits the growth of other
alcohol) or as an iodophor (combined with an organic
microorganisms.
molecule). The germicidal action of chlorine is based on
- Comes from the word antibiosis
the formation of hypochlorous acid when chlorine is added
- Naturally derived
to water. Chlorine is used as a disinfectant in gaseous
form (C12) or in the form of a compound, such as calcium Antibiosis - refers to the different ways a
hypochlorite, sodium hypochlorite, and chloramines. microorganism kills other microorganism
- Interaction between organisms

UNIT VI: Antimicrobial Drugs

The History of Chemotherapy


Introduction
Early Humans have known that there are small
Selective toxicity: A drug/chemical that kills harmful organisms everywhere
microbes without damaging the host
- Toxic only to certain types of cells Germ theory of disease - there are microorganisms that
- Needs to distinguish between different types of can cause diseases.
cells
Before the discovery of antimicrobial agents, if harmful
microorganisms managed to get into your body then there
is a likelihood that you will die because of that
microorganism (fatal).

Several scientists wanted to create medication or


chemicals that can enter our body, but do not cause harm
to the human body, but can still kill bacterial cells.
(selective toxicity).

Paul Ehrlich developed the concept of chemotherapy to Sulfanilamide - Derived from a dye
treat microbial diseases.
- First commercially available antimicrobial drug
- Father of Chemotherapy
- One of the few scientists that wanted to find a - Marketed Sulfanilamide as (Prontosil)
treatment for treponema pallidum infection - One of the first medications to arrive in the
(Syphilis) that can affect your genitals and even philippines
the CNS (Neurosyphilis). - Not an antibiotic
- Tried 606 Compounds for syphilis
- Compound 606 (Salvarsan) an arsenic
compound capable of killing treponema pallidum
without killing healthy cells
- Compound 606 (Salvarsan) is the first selective The History of Chemotherapy
toxicity drug.
- Also known as Arsphenamine or Magic Bullet Alexander Fleming discovered the first
antibiotic,penicillin,in 1928; its first clinical traits were done
Compound 606 (Salvarsan) - contains harmful in 1940.
metalloids that can produce adverse drug reaction which - Accidentally discovers penicillin while creating
is no 100% safe culture media
- Stepping stone for future selective toxicity drug - Penicillin came from penicillium notatum or the
- Proved that selective toxicity is possible mold in his culture media
- It is not an antibiotic (synthesized in a lab) - Was about to discard the culture sample but was
helped by two scientist to create the treatment

Howard Florey & Ernst Chain - The two scientist that


help fleming develop penicillin as a medication
- Created and develop tests in order to manufacture
penicillin

Note: The use of Antiseptics, Disinfectants, and


Heating is only applicable when microorganisms are
outside the body, because humans are susceptible to heat
(causes our proteins to coagulate).

High alcohol concentration should be used in disinfecting,


but should not be ingested or taken orally because it is Note: Penicillin is an antibiotic
harmful (we metabolize alcohol which is toxic).

The History of Chemotherapy


The History of Chemotherapy
Most antibiotics are made by Streptomyces bacteria.
Sulfa drugs came into prominence in the 1930’s.
- Eli Lilly
- Erythromycin (Ilosone)
Gerhard Domagk was a German pathologist and
bacteriologist credited for the discovery of Sulfanilamide
(Prontosil). Became the first commercially available
antimicrobial drug.
- Tried different chemicals like Paul Ehrlich to treat
his daughter
- Successfully treated a patient with Sulfanilamide
- Received a nobel peace prize in physiology for his
discovery of Sulfanilamide
cells.
Note: Dr. Dalisay - Filipino scientist currently - Fungi and protozoa have their own
gathering soil samples from the bottom of the ocean eukaryotic enzymes.
inorder to get microbial sample to create new -
- The more similar the pathogens and host
antibiotics
enzymes, the more side effects the
antimicrobial will have.

Action of Antimicrobials

The Spectrum of Activity


Antibacterial drugs affect many targets in a prokaryotic
cell.
Fungal, protozoan, and helminthic infections are more
difficult to treat because these organisms have eukaryotic
cells.
Narrow-spectrum drugs affect only a select group of
microbes (grams-positive cells,for examples); broad
spectrum drugs affect a more diverse range of microbes.

The Action of Antimicrobial Drugs

Antimicrobials generally act either by directly killing


microorganisms or by inhibiting their growth.

- cidal: Chemical that kills microorganisms


● Bactericidal/Fungicidal
-static: Chemical that inhibits the growth of
microorganisms
● Bacteriostatic/Fungistatic

The Spectrum Activity

- Small,hydrophilic drugs can affect


gram-negative cells.
- Antimicrobial agents should not cause
excessive harm to normal microbiota.
- Superinfections occur when a
pathogen develops resistance to the
drug being used or when normally
resistant microbiota multiply
excessively. Cell wall of prokaryotes

The Action of Antimicrobial Drugs

Bacteria have their own enzymes for:


- Cell wall formation
- Protein synthesis
- DNA replication
- RNA synthesis
- Synthesis of essential metabolites

The Action of Antimicrobial Drugs Inhibitors of Cell Wall Synthesis

- Viruses use host enzymes inside host


Penicillins synthesis and may be used to kill penicillinase
All penicillins (over 50 compounds) contain a 4 sided ring producing staphylococci.
or β-lactam ring.
Natural penicillins produced by Penicillium spp. Are Note: Bacillus subtilis strain was gathered from Margaret
effective against gram-positive cocci and spirochetes. Tracey

Inhibitors of Cell Wall Synthesis

Antitubercular Drugs
Isoniazid (INH) and ethambutol inhibit cell wall synthesis
in mycobacteria by interfering with mycolic acid synthesis
or incorporation.

Inhibitors of Cell Walls


Inhibitors of Protein Synthesis
Semisynthetic penicillins are resistant to penicillinase Chloramphenicol,aminoglycosides,tetracyclines,
and have a broader spectrum of activity than natural glycylcyclines,
penicillins. macrolides,streptogramins,oxazolidinones,and
pleuromutilins inhibit protein synthesis at 70s ribosomes.
Carbapenems are broad-spectrum antibiotics that inhibit
cell wall synthesis.

The monobactam Aztreonam affects only gram-negative


bacteria. Injury to Membranes

Lipopeptides

Polymyxin B and Bacitracin cause damage to


plasma membranes.

Nucleic Acid Synthesis Inhibitors

Rifamycin
Inhibits mRNA synthesis; it's used to treat
Inhibitors of Cell Wall Synthesis tuberculosis.
- First treatment for TB
Cephalosporins - Inhibit cell wall synthesis and - Antibiotic
are used against penicillin resistant strains.
- Derived from Streptomyces griseus
2nd,3rd and 4th generations are more effective - Discovered by Selman Waksman
against gram-negative bacteria.
R I P E S (Rifampicin, Isoniazid, Pyrazinamide,
Polypeptide - Such as Bacitracin inhibit cell wall Ethambutol, Streptomycin) - effective cure for TB
synthesis primarily in gram-positive bacteria.

The glycopeptide Vancomycin inhibits cell wall Quinolones


Quinolones and fluoroquinolones, such as synthesis or protein cutting in virus assembly.
Ciproflaxin, inhibit DNA gyrase.

Antimetabolites

Sulfonamides
Competitively inhibit the synthesis of essential
metabolites such as folic acid.

SMZ-TMP competitively inhibits dihydrofolic acid


synthesis.

- Sulfamethoxazole + Trimethoprim (Co-


Trimoxazole)

- Entry inhibitors and fusion inhibitors bind to


viral attachment and receptor sites. Inhibit
attachment:
• Zanamivir (tx of influenza)

Inhibit uncoating:
ANTIFUNGAL DRUGS • Amantadine (tx of influenza)

- Pathogenic fungi are often outside the body. Nucleoside and nucleotide analogs, such as acyclovir
and zidovudine, inhibit DNA or RNA synthesis.
- Fungi are eukaryotes and have unique sterols
(ergosterol) in their cell walls.

- Polyenes, such as nystatin and amphotericin


B, combine with plasma membrane sterols
and are fungicidal.

- Azoles and allylamines interfere with sterol


synthesis and are used to treat cutaneous
and systemic mycoses.
Inhibitors of viral enzymes prevents viral assembly and
exit. Inhibits assembly:
- Echinocandins interfere with fungal cell wall
synthesis. • Indinavir (tx of HIV)
- The antifungal agent flucytosine is an
antimetabolite of cytosine. Alpha interferons are natural products of the immune
system in viral infections. They inhibit the spread of
- Griseofulvin interferes with eukaryotic cell viruses to new cells. (tx of Viral hepatitis)
division and is used primarily to treat skin
infections caused by fungi.

ANTIPROTOZOAL DRUGS

Protozoa are eukaryotic cells.


ANTIVIRAL DRUGS Many drugs are experimental, and their mode of action is
unknown.
- Viruses are composed of nucleic acid, protein
Chloroquine, artemisinin, quinacrine, diiodohydroxyquin,
capsid, and host membrane containing virus
pentamidine, and metronidazole are used to treat
proteins.
protozoan infections.
- Viruses live inside host cells and use many host
enzymes.
Metronidazole — inhibit glucose uptake; tx of
- Some viruses have unique enzymes for DNA/RNA
• Trichomonas vaginalis E-TEST
• Entamoeba histolytica
• Giardia lamblia Minimal inhibitory concentration (MIC) is the lowest
• Anaerobic bacteria concentration of drug capable of preventing microbial
growth; MIC can be estimated using the E-test.

ANTHELMINTIC DRUGS

Helminths are macroscopic multicellular eukaryotic


organisms: tapeworms,roundworms,pinworms or
hookworms.

Anthelmintic drugs include mebendazole, praziquantel and


ivermectin.

- Prevent ATP generation (Tapeworms)


- Alters membrane permeability (Flatworms)
- Neuromuscular block (Intestinal roundworms)
- Inhibits nutrient absorption (Intestinal
roundworms)
- Paralyzes worm (Intestinal roundworms)

TESTS TO GUIDE CHEMOTHERAPY BROTH DILUTION METHODS BROTH DILUTION


TESTS

TESTS TO GUIDE CHEMOTHERAPY - In a broth dilution test, the microorganism is grown


in liquid media containing different concentrations
Measuring Antimicrobial Sensitivity of a chemotherapeutic agent.
- MIC and MBC can be determined using the broth
- Tests are used to determine which dilution method.
chemotherapeutic agent is most likely to combat a - The lowest concentration of a chemotherapeutic
specific pathogen. agent that kills bacteria is called the minimum
- These tests are used when susceptibility cannot bactericidal concentration (MBC).
be predicted or when drug resistance arises.
• Disk-diffusion tests
• Broth dilution tests

Serial Dilutionp

KIRBY-BAUER TEST

In the disk-diffusion test, also known as the KirbyBauer


test, a bacterial culture is inoculated on an agar medium,
and filter paper disks impregnated with chemotherapeutic
agents are overlaid on the culture.

Antimicrobial Concerns

After incubation, the diameter of the zone of inhibition is RESISTANCE TO ANTIMICROBIAL DRUGS
used to determine whether the organism is sensitive,
intermediate, or resistant to the drug. - Many bacterial diseases, previously treatable with
antibiotics, have become resistant to antibiotics.
Antimicrobial Resistance
• Relative or complete lack of effect of antimicrobial
against a previously susceptible microbe
• Increase in MIC
Superbugs are bacteria that are resistant to several
antibiotics.

Antibiotic present: Resistance to Antimicrobial Drugs


Some combinations of drugs are synergistic; they are
Drug resistance factors are transferred horizontally more effective when taken together
between bacteria.
Some combinations of drugs are antagonistic; when taken
together, both drugs become less effective than when
taken alone

Synergistic effect - drugs that are more effective taken


together
Ex: Co-trimoxazole

Additive Effect - effects of two drugs

Antagonistic effect - Better to be taken separately or will


lose effectiveness
RESISTANCE TO ANTIMICROBIAL DRUGS Ex:
MECHANISMS OF ANTIBIOTIC RESISTANCE Potentiation Effect - there is one effective against
1. Enzymatic destruction of drug microorganism, and another drug with no antimicrobial
2. Prevention of penetration of drug effect that creates a stronger antimicrobial activity
3. Alteration of antibiotic or target site Ex: amoxicillin + Clavulanic acid = Stronger antimicrobial
4. Rapid ejection of the drug activity

Combination

Additive: 1 + 1 = 2
Synergistic: 1 + 1 = 3
Antagonistic 1 + 1 = 0
Potentiation 1 + 0 = 2

FUTURE OF CHEMOTHERAPEUTIC AGENTS

- New agents include antimicrobial peptides,


bacteriocins, and bacteriophages.
The discriminating use of drugs in appropriate
concentrations and dosages can minimize resistance. - Virulence factors rather than cell growth factors
may provide new targets.

ANTIBIOTIC SAFETY

The risk (e.g., side effects) versus the benefit (e.g., curing UNIT VII: Principles of Disease and Epidemiology
an infection) must be evaluated before antibiotics are used

Normal Microbiota of Humans


EFFECTS OF COMBINATIONS OF DRUGS
- Internal organs are usually free of
microorganisms.
- But surface tissues have extensive populations of
microorganisms.
- The microbes that normally inhabit a healthy
individual's body are called microbiota or "normal
flora". Microbiota are specialists, able to colonize
and survive on human tissue.
- Each body region has characteristic flora. Symbiosis

Skin Symbiosis is when two organisms live intimately close,


typically over longer periods, often measured in
- Not a great habitat dries out, constantly being generations for at least one of the organisms. aIf one
shed, secretions include fatty acids (lower pH to 4- organism is substantially smaller than the other organism
6) and saltSome skin regions better habitats than and lives in or on the larger, then the larger organism is
others a scalp, ears a underarms anal region referred to as a host and the smaller as a symbiont.
- Propionibacterium acnes can live in sweat glands,
hair follicles, so it is not eliminated by washing - Mutualism is a symbiosis in which both host and
skin symbiont benefit.
- Commensalism is a symbiosis in which one of the
- Staphylococcus epidermidis and Staphylococcus participants (typically the symbiont) benefits but
aureus, are found on skin and thrive in nasal the other organism (typically the host) neither
region benefits nor is harmed.
- Parasitism, the third category of symbiosis, is one
Mouth where the host is harmed while the symbiont gains
(the latter, e.g., by having a place to live and
- Saliva contains lysosomes and other enzymes something to eat).
that kill bacteria.
- Streptococcus mutans and other Streptococcus
adheres to teeth, especially gum margins, Host-Microbe Relationship
providing microhabitat for other bacteria to
colonize. Microbial competition
It is the prevention of harmful bacterial growth by a non
Gl tract harmful bacteria.

- Stomach is highly acidic (pH 2-3) and kills most - An organism existing in an area on or in the host
microbes. fills the area and thus serves to prevent the filling
- Some bacteria and yeasts can tolerate passage of the same area by a harmful bacterium.
through stomach; few microorganisms live in - Normal microbiota can benefit the host by
stomach. preventing overgrowth of harmful microorganisms.
- other words, the good bacteria help defend
- Small intestine has some bacteria, but does not against the bad bacteria.
proliferate due to digestive enzymes.
- As it approach colon, more and more bacteria can Resident microbiota
be seen, especially Gram-negative Resident microflora are the more-or-less permanent
- Enterobacter (e.g. Escherichia coli). aColon has members of normal microflora.
an enormous bacterial population (1/3 of feces is
bacteria). Transient microbiota
- Bacteria in colon divide every 12-24 hours on Transient microflora are present only under unusual
average, much slower than laboratory batch circumstances and only transiently present (hours to
culture rates. months).
Genitourinary tract
Opportunistic microorganisms
- Upper urinary tract is usually sterile. Opportunists are members of the normal microflora that do
- Lower part of the urethra gets some bacteria, but not usually cause disease but can be pathogenic under
is frequently "washed out" by urinary flow. certain circumstances like:
- When washout is reduced, more chances of ● Host immunosuppression
urinary tract infections. ● Transfer to other parts of the body
- Vagina has complex microbiota. After women start ● Elimination of microbial antagonism
periods, glycogen is secreted, and lactic acid
bacteria produce lactic acid, maintaining pH 4.5.

- The good bacteria defend against the bad bacteria


by: aCompeting for attachment sites aCompeting Pathology
for nutrients aMaking antibiotics against invading
Pathology
microbes
Study of disease aConcerned with etiology as well as
- Disease occurs when this balance between the
structural and functional changes brought about by the
normal flora and the host is upset (e.g., antibiotic
disease.
misuse).
- An infection that results from a prior infection is
Infection
called a secondary infection
Invasion and colonization of the body by pathogenic
microorganisms. used by microorganisms to adhere to and colonize body
surfaces.
Disease
Occurs when an infection results in any change from a - Mechanical means such as suckers, hooks, and
state of health. An abnormal state in which part or all of barbs are used by parasites, such as tapeworms,
the body is incapable of performing normal functions. to adhere and colonize body surfaces.

Pathogenicity Becoming Established


An organism's capacity to cause disease. Once the organism has made it into the body and attached
it still has to avoid or defend itself from the body's internal
Virulence defenses, the immune system.
The degree of disease an organism has the potential to
cause disease. Growing the pathogen under conditions Capsules or other disguises are used by microorganisms
that decrease its adaptation to growth on a given host will to avoid phagocytosis.
decline the virulence of pathogen (attenuation). aSome microorganisms make chemicals that are toxic to
white blood cells called leukocidins

Events of Infection

Becoming Established: Surviving Host Defenses


There are 3 main factors that aid a microbe in becoming
established.

A. It enters or gains access to the body through the


correct portal of entry. Stages of Disease
B. The number of cells that enter the body is enough
to escape the body's defenses. The period during which the disease is spread is
C. Other predisposing dependent upon the infecting organism.
For example, some organisms are contagious before
Portal of Entry signs and symptoms are manifest. Others are contagious
These are the ways to enter the host. Microbes can't during the worst portion of the manifest signs and
cause disease unless they enter the body through the right symptoms.
opening.
Syndrome
3 main ways that microbes enter the body A combination of signs and symptoms that are
characteristic of a disease.
1. Mucous membranes: Respiratory tract (RT),
Gastrointestinal tract (GIT), Urogenital tract (UT), Symptom is a characteristic of a disease that can be felt
conjunctiva. (by the individual with the disease) but cannot be
2. Skin: unbroken, impenetrable to most people. measured by another individual. aSign is a characteristic
Can gain access through hair follicles, sweat of a disease that can be measured by another individual
ducts, abrasions
3. Parenteral route: deposited directly into tissues
beneath skin aPunctures, injections, cuts, wounds,
surgery, cracking

Size of Inoculum
Number of cells that enter through the portal of entry aID
(Infectious Dose): minimum number of cells required to
cause infection. Some organisms require 100's of cells to
cause infection while others require as few as 10.

Predisposing Factors
One that makes the body more susceptible to a disease Incubation period
and may alter the course of the disease. c] Gender - Time interval between the initial infection and the
aClimate and weather aOthers include nutrition, age, first appearance of any signs or symptoms.
fatigue, etc. - The time an infection has begun up to the
occurrence of signs and symptoms.

Attachment (issues) Prodromal period


- Short period that follows the period of incubation
After the microbe has entered the body, the next step is to in some diseases when symptoms (and signs)
attach and colonize the body surfaces. appear, but full blown illness has not-yet begun
Adhesion: fimbriae, surface proteins, and capsules are - Early, mild symptoms
Are toxins associated with Gram-negative bacteria
Period of Illness
- The phase during which the typical signs and - The lipid A portion of the lipopolysaccharide
symptoms of the disease are apparent. aWeak except in large doses and produce similar
- Most acute stage of the illness c] Number of white effects independent on the producing organism
cells increases or decreases - Large doses are especially a problem during
Peak of Illness Gram Negative septicemia
- The peak of disease symptoms aThe person - Cause damage that is a consequence of the body
exhibits full signs and symptoms of the disease. using endotoxins as a signal for Gram-negative
bacterial infections; excessive amounts of
Period of Decline endotoxin cause the body to overreact and
- the period during which the signs and symptoms damage itself.
subside as the infection is brought - Typically, are released following bacterial division
- further under control or lysis.
- Patient is susceptible to secondary infections - There is no such thing as an endotoxin toxoid nor
a vaccine against endotoxin
Period of Convalescence
- Person regains strength and the body returns to
its pre diseased state aSequelae are persisting
disease after infection Kinds of Infectious Diseases
- The inability of the body to fully repair the damage
due to an infection A. Kinds of Infectious Diseases

● Infectious disease is a disease that involves


Virulence Factors pathogens.
● Communicable disease is an infectious disease
Virulence factors that may be passed from host to host (particularly
properties of pathogens that allow them to cause disease. when all individuals involved are of the same
Any characteristic or structure of the microbe that helps it species)
to establish itself in the host or cause damage in the host. ● Contagious disease is easily passed from
- Pili are used in adhesion to hosts aCapsules individual to individual.
prevent phagocytosis
- Enzymes used to evade host defenses or to harm B. Types of Infection
the host directly ● An acute infection develops rapidly but is soon
- Toxins are substances produced, for example, by over
microorganisms, that are poisonous to host ● A chronic infection develops slowly and is not
organisms soon over
● A subacute infection is the gray zone between
Bacterial toxins may be classified as either exotoxins or acute and chronic
endotoxins ● A latent infection is sign-less or symptom-less for
a long while before signs and symptoms appear
Exotoxins are produced predominantly (though not ● An inappropriate infection that does not display
exclusively) by Gram-positive bacteria aAct by a variety of signs or symptoms or, at least, all of the signs
mechanisms and symptoms that result from exposure to typically associated with a given syndrome.
an exotoxin depending on the structure of the exotoxin ● A local infection is confined to a certain area (e.g.,
one is exposed to. a pimple).
- Made inside a living microbe and released into the ● A focal infection begins as a local infection but
surrounding environment. a Can be converted to then spreads beyond the local area as a
toxoid bacteremia, or toxemia.
- Some exotoxins are exoenzymes ● A systemic infection is spread throughout the body
- Hemolysins that catalyze the lysis of red blood in the blood or lymph.
cells Mucinase, keratinase, collagenase,
hyaluronidase C. Blood Invasion
- Other exotoxins damage or destroy phagocytic ● Septicemia is the growth of bacteria in the blood
cells that have engulfed an exotoxin producer. ● (a.k.a., blood poisoning)
- Named according to the mode of action or the ● Bacteremia is the presence, without multiplication,
specific tissues they act against. of bacteria in the blood.
- Cytotoxins kill host cells by damaging the cell ● Fungemia is the presence of fungus in the blood
membrane. ● Toxemia is the presence of toxins in the blood
- Neurotoxins act on nervous system tissue ● Viremia is the presence of virus in the blood

Enterotoxins act on tissues of the gastrointestinal tract

Endotoxin
Koch’s Postulates
The means, or logic, by which a specific Common Infectious Diseases and Causative
microorganism is classified as the cause of a Microorganisms in the Nervous System
disease.

1. Evidence that the causative agent is always


present when the specific disease is present.
2. Successful isolation of the causative agent in
pure culture.
3. Ability of pure-cultures of the organism to
cause disease in a healthy organism.
4. Successful isolation of the causative agent
from the experimentally infected host.

Exceptions to Koch's postulates Common Infectious Diseases and Causative


Microorganisms in the Genitourinary Tract
1. There may exist more than one cause of a
specific disease.
2. It might not be possible to grow the causative
agent in pure culture.
3. There might not exist a suitable experimental
host.
4. The disease might result from more than one
causative agent acting in unison.

Common Infectious Diseases and Causative


Microorganisms in the Respiratory Tract
Basic Immunology

Definitions:
- Immune System = cells, tissues, and molecules
that mediate resistance to infections.
- Immunology = study of structure and function of
the immune system.
- Immunity = resistance of a host to pathogens and
their toxic effects.
- Immune response = collective and coordinated
response to the introduction of foreign substances
in an individual mediated by the cells and
molecules of the immune system.

Common Infectious Diseases and Causative Role of the Immune System


Microorganisms in the Gastrointestinal Tract - Defense against microbes
- Defense against the growth of tumor cells
- Kills the growth of tumor cells
- Homeostasis
- Destruction of abnormal or dead cells
(e.g. dead red or white blood cells, antigen-antibody
complex.

Immune System
1. Organs
2. Cells
3. Molecules

Immune System:
(1) organs Innate Immunity: Mechanisms
- Mechanical barriers/surface secretion
- Skin, acidic pH in stomach, cilia
- Humoral Mechanisms
- Lysozymes, basic proteins, complement,
interferons.
- Cellular defense mechanisms
- Natural killer cells, neutrophils, macrophages,
mast cells, basophils, eosinophils.

Immune System:
(2) cells
- Lymphocytes
- T-lymphocytes
- B-lymphocytes, plasma cells
- Natural killer lymphocytes
- Monocytes, Macrophage
- Granulocytes
- Neutrophils
- Eosinophils
- Basophils

Immune System:
(3) molecules
- Antibodies
- Complement
- Cytokines
- Interleukins
- Interferons

Two types of Immunity:


1. IInnate (non-adaptive)
- First line of immune response
- Relies on mechanisms that exist before
injection.
2. Acquired (adaptive)
- Second line of response (if innate fails)
- Relies on mechanisms that adapt after
infection
- Handled by T and B-lymphocytes
- One cell determines one antigenic
determinant.

Innate Immunity:
First line of defense
- Based on genetic make-up
- Relies on already formed components
- Rapid response: within minutes of infection Adaptive Immunity:
- Non-specific Second line of response
- Same molecules/cells respond to a range of - Based upon resistance acquired throughout life
pathogens - Relies on genetic events and cellular growth
- Has no memory - Responds more slowly, over few days
- Same response after repeated exposure. - Is specific
- Does not lead to clonal expansion. - Each cell responds to a single epitope on an
antigen
- Has anamnestic memory
- Repeated exposure leads to faster, stronger
response.

Adaptive Immunity: Active and Passive

Adaptive Immunity:
Mechanisms
- Cell mediated immune response (CMIR) 1. T-cell
- T-lymphocytes - Recognizes peptide antigen on
- Eliminate intracellular microbes that survive macrophage in association with major
within phagocytes or other infected cells histo-compatibility complex (MHC) class
- Humoral Immune Response (HIR) - Identifies molecules on cell surfaces.
- B-lymphocytes - Helps body distinguish non-self materials
- Mediated by antibodies 2. T-cell goes into effectors cells stage that is able to
- Eliminate extra-cellular microbes and their toxins kill infected kills.

Humoral Immune Response


1. B lymphocytes recognize specific antigens.
- Proliferate and differentiate into antibody-
secreting plasma cells. T lymphocytes
2. Antibodies bind to specific antigens on 2 types
microbes via specific mechanisms. - Helper T-lymphocytes (CD4+)
3. Some B lymphocytes evolve into the resting - CD4+ T cells activate phagocytes to kill microbes
state - Cytolytic T-lymphocyte (CD8+)
- Memory cells - CD8+ T cells destroy infected cells containing
microbes or microbial proteins.

Cell mediated immune response


- Primary response
- Production of specific clones of effector T cells
and memory clones
- Develops in several days
- Does not limit the infection
- Secondary response
- more pronounced, faster
- more effective at limiting the infection

Example - cytotoxic reactions against intracellular


parasites, delayed hypersensitivity (e.g, Tuberculin test)
and allograft rejection.

Antibodies (immunoglobulins)
- Belong to the gamma-globulin fraction of serum lgG
proteins • 70-75% of total immunoglobulin
- Y-shaped or T-shaped polypeptides • Secreted in high quantities in secondary
- 2 identical heavy chains exposures
- 2 identical light chains • Cross the placenta
- Five kinds of antibodies • Major functions /
- lgG, lgM, lgA, lgD, lgE applications
• neutralize microbes and
toxins
• opsonize antigens for phagocytosis
activate the complement
• protect the newborn
• participate in agglutination and precipitation reactions

• 4-fold rise or fall indicates active infection


• A single positive sample indicates past exposure

lgM
• Secreted initially during primary infection
• Major functions / applications
• secreted first during primary exposure
• activates the complement
• used as a marker of recent infection
Immunoglobulin

•Presence in newborn means infection


•Single positive sample in serum or CSF indicates
recent or active infection
•Used to detect early phase of infection

lgA
• Monomeric in serum (lgA1)
• Dimeric with secretory component in the lumen of the
gastro-intestinal tract and in the respiratory tract (lgA2)
• Major function / application
• neutralizes microbes and toxins

•Sero-diagnosis of tuberculosis
lgM - lgG sequential response
•Synthicial respiratory virus tests

lgD
• Monomeric
• Extremely scarce (<0.2% of total immunoglobulin)
• Major functions / applications
• present on the surface of B lymphocytes
• functions as membrane receptor
• role unclear
• has a role in antigen stimulated lymphocyte
differentiation

lgE
• Mediates type I hypersensitivity
• Monomeric
• Major functions / applications Immunoglobulin
• associated with anaphylaxis
• plays a role in immunity to helminthic parasites

Serodiagnosis of infectious and non-infectious allergies


(e.g., allergic bronchopulmonary aspergillosis, parasitic
diseases)

Antigens and immunogens

Antigen
A substance that reacts with antibody molecules and
antigen receptors on lymphocytes

Immunogen
An antigen that recognized by the body as non-self and
stimulates and adaptive immune response

For simplicity, both antigens and immunogens are usually


referred to as antigens

To be immunogenic, an antigen must possess three


characteristics:
IgM
A. High molecular weight
● Produced as a first response to many antigens B. Chemical complexity
● Levels remain high transiently C. Foreignness (recognized as nonself by the body)

Chemically, Antigens are large molecular weight proteins


IgG and polysaccharides. Including conjugated proteins such
as glycoproteins lipoproteins and nucleoproteins and
● Produced after IgM including lipopolysaccharide
● Higher levels persist in small amounts throughout
life
● Produced in large amounts during secondary Epitopes
response Epitopes (or antigenic determinants)

Antigens and immunogens:


• Type 2-Cytotoxic
• the actual portions or fragments of an antigen that • Type 3-Immune Complex
react with receptors on B-lymphocytes and • Type 4- Delayed
Tlymphocytes, as well as with free antibody
Molecules

• Composed of polysaccharides or proteins recognition Hypersensitivity types


• The body recognizes an antigen as "foreign" when
epitopes of that antigen binds to Blymphocytes and T-
lymphocytes

• By means of epitope-specific receptor

Antigens and immunogens


Haptens
A hapten is a small molecule that by itself is not
immunogenic acts as an antigen when it binds to a larger
protein molecule

Antigens and immunogens

Immunodeficiency

● Loss or inadequate function of various


components of the immune system

● Can occur in any part or state of the immune


system

● Physical barrier, phagocytes, B lymphocytes, T


lymphocytes, complement, natural killer cells

Failure of immune response ● The immuno-compromised host

Immune response helps individuals defend against: ● has an impaired function of immune system

• microbes ● is at high risk of infection


• some cancers
• Immune response can fail ● Congenital (primary) immunodeficiency
• hypersensitivity reactions
• immunodeficiency ● Genetic abnormality

● Defect in lymphocyte maturation


Hypersensitivity reactions
● Acquired (secondary) immunodeficiency
Cause cell damage through excessive immune response
to antigens: ● Results from infections, nutritional deficiencies or
treatments
• Hypersensitivity
• overreaction to infectious agents ● AIDS, chronic leukemia
• Allergy
• overreaction to environmental substances
• Autoimmunity
• overreaction to self Altered immunity: immunocompromised

Hypersensitivity reactions

• Type 1 -Anaphylaxis

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