BACTERIAL GROWTH

AND
CULTIVATION
WEEK 2
MICROBIAL NUTRITION
Microorganisms require specific nutrients that are essential for their growth and
multiplication. Examples are the oxygen, carbon, hydrogen, nitrogen and sulfur.
These are some of the requirements needed in energy production and biosynthesis.
 ◦ ACCORDING TO OXYGENT REQUIREMENTS
1. Aerobes
- These organisms require oxygen to grow well with room air.
- Air contains 15-21% oxygen and 1% CO2.
- Examples are the Bordetella, Brucella, Mycobacteria & Pseudomonas.
2. Anaerobes
PHYSIOLOGIC
- These organisms do not require oxygen to grow.
REQUIREMENTS
Different types of Anaerobes.
OF BACTERIA
a) Obligate anaerobes
- They do not require the presence of oxygen because they die
after prolonged exposure to air.
- Examples are Clostridium and Bacterioides.
b). Facultative anaerobes

- These are the most clinically significant bacteria.

- Organisms can either grow in the presence or absence of oxygen.

- These are aerobes that can grow anaerobically.

- These organisms do not require oxygen but grow better in the presence of it.

- Examples are the bacteria under Enterobacteriaceae.

c). Aerotolerant anaerobes

- Organisms can survive in the presence of oxygen but will be unable to perform metabolic process unless situated in an
anaerobic environment.

- Example is Propionibacterium acnes

3. Microaerophile

- It requires 2-10% oxygen for growth

- Some examples are Campylobacter and Treponema pallidum

◦ ACCORDING TO CARBON DIOXIDE REQUIREMENTS

Capnophiles require and increased CO2 (5%-10%)

Some examples of Capnophiles are Haemophilus influenzae, Neisseria gonorrhea, and Streptococcus pneumoniae

ACCCORDING TO NUTRITIONAL REQUIREMENT

1. As to carbon source
- AUTOTROPHS - It uses CO2 as the sole source of Carbon.

- HETEROTROPHS – It uses reduced, performed organic molecules from other bacteria.

2. As to energy source
- PHOTOTROPHS – It uses Light as energy source

- CHEMOTROPHS – It utilizes the energy produced by organic or inorganic compound oxidation.

3. As to the electron source
- LITOTHROPHS – Reduced inorganic molecules to be used in biosynthesis or energy
conservation.
- ORGANOTHROPHS – It require organic substances ( Protein, Lipids) for growth and
multiplication.

ACCORDING TO TEMPERATURE REQUIREMENT

- The optimum temperature for most of the bacteria to grow is 35°C to 37°C
1. Psychrophiles/Cryophiles
- These organisms grow well at 0 °C to maximum 20°C
- Examples are Listeria monocytogenes and Yersinia enterolitica.
2. Mesophiles
- These organisms grow between 20°C to 45 °C
- Most commonly encountered pathogenic bacteria in clinical laboratory.
3. Thermophiles/ Hyperthermophiles
- These organisms grow between 50°C to 125 °C
4. Extremophiles
- These are prokaryotes that can survive in unusual conditions like in the absence of oxygen, increased temperatures, and
living below the earth’s surface.
- Example: Bacillus infernus.
THERMAL DEATH TIME – lowest/minimum time required to kill organisms under constant temperature.
THERMAL DEATH POINT- lowest temperature required to kill organism in a constant time.

ACCORDING TO pH REQUIREMENT
The optimum pH for most pathogenic bacteria is from pH 6.5-7.5.
1. Acidophiles- they grow between pH 0-5.5
2. Neutrophiles- they grow between pH 5.5-8.0 (most clinically significant bacteria)
3. Alkalophiles – they grow between pH 8.5-11.5
ACCORDING TO MOISTURE
- Moisture is important for bacterial growth and susceptibility testing.

ACCORDING TO SALT CONCENTRATION

1. Halophile
- They require increased concentration of sodium chloride.
- Examples: Staphylococcus aureus and Listeria monocytogenes
ACCORDING TO PRESSURE
1. Barophiles
- They grow rapidly in increased pressure environment.
- Examples: Photobacterium, Shewanella, Collwellia.
ACCORDING TO GROWTH FACTORS
- Growth factors are substances required by fastidious bacteria for their growth and
multiplication. Examples are the purines, amino acids, pyrimidines and vitamins.
BACTERIAL GROWTH CURVE

STATIONARY
PHASE

LOG PHASE
DECLINE
PHASE

LAG PHASE
◦ Terminology
1. Generation- doubling of cell number.
2. Generation time/ Doubling time- time required for bacteria to double their population.

STAGES OF BACTERIAL GROWTH

1. Lag phase/ Period of rejuvesence
- The period where there is no cell division.
- The start of biosynthesis.
- Adjustment phase of bacteria.

2. Log phase or Exponential phase

- The period where there is actively growing and actively dividing bacteria.
- It is the best stage to which microorganisms are utilized in physiological and biochemical testing.
3. Stationary phase/ Plateau phase
- Period where there is a balance between cell division and dying.
4. Death phase/ decline phase
- Period where there is cessation of bacterial growth as number of death cells exceeds the number of living organisms.

BACTERIAL GROWTH MEASUREMENT

1. By cell count- done through microscope, electronic particle counter, or colony count.
2. By cell mass- done by weighing, measuring the nitrogen content and turbidimetry.
3. By cell activity- done by relating biochemical activity.
 BACTERIAL COLONY
◦COLONIES ARE VISIBLE MACROSCOPIC MASSES OF
GROWTH ON A SOLID CULTURE MEDIUM AS A
RESULT OF REPEATED DIVISIONS OF ONE OR
FEW MICROORGANISMS
 EVALUATION OF COLONY
◦ laboratorians can provide physicians with early preliminary information regarding the patient’s culture result.

◦ This information also is important for deciding which subsequent steps to take for definitive organism
identification and characterization.

a) Type of media supporting the bacterial growth.

b) Incubation condition.

c) Relative quantities of each colony type.

d) Colony characteristics.
Criteria frequently used to characterized
bacterial growth include the ff:
◦Colony size
◦Colony pigmentation
◦Colony shape
◦Colony surface appearance
◦Changes in agar media resulting from bacterial growth.
◦Odor
MUCOID
COLONY
◦ It exhibits a water-like glistening, confluent
appearance
◦ Characteristic of organisms that forms slimes
or well developed capsules.
◦ Example: Klebsiella pneumoniae, S. pneumoniae, H.
influenzae
(ALL ENCAPSULATED BACTERIA)
SMOOTH
COLONY
◦ It has uniform texture and
homogeneity.
◦ It easy emulsifies in NSS
◦ Examples: Salmonella, Shigella, E. coli,
Serratia, Proteus
ROUGH
COLONY
◦ Granulated and rough in appearance
◦ It is usually produced by mutant strains that
lack surface proteins or polysaccharides
indicating loss of virulence.
 METHODS FOR MEASURING
BACTERIAL GROWTH
1. Microscopic count

- A measured volume of bacterial suspension is placed inside a defined area on a microscopic slide.

2. Plate count

- Most commonly used method.

- It measures the number of viable cells.

- It determines the CFU/mL of bacteria.

- 30-300 colonies should be counted.

3. Membrane or molecular filter

- It utilizes a polycarbonate membrane filter or Millipore filter.

4. Turbidimetric method ( Cell mass)

- It requires 10-100 million cells/Ml

5. Dry weight determination ( Cell mass)

-for filamentous organisms

6. Biochemical activity

- Enumeration of bacteria in milk.

 CULTURE AND CULTURE MEDIA
◦ Cultures are the growth of microorganisms in a culture medium.

◦ It relies on their ability to grow and survive outside the host, use of culture methods is certainly efficient.

◦ A culture medium is composed of a mixture of nutrients such as carbon, nitrogen, oxygen, sulfur, phosphorus,
hydrogen and buffers.

◦ Inhibitory agents like dyes, salts, and antimicrobials are added into the medium to facilitate the isolation of the
desired organism while suppressing the growth of other organisms present in sample.

◦ The selection of medium for inoculation is based on the type of specimen submitted for culture and the kind
of organisms that are involved in the infection process.
TYPES OF
CULTURE
◦ PURE CULTURE
- It is composed of only one species.
◦ MIXED CULTURE
- It is composed of more than one species.
◦ STOCK CULTURE
- It is composed of several species contained in a
separate culture medium; one specie per culture
medium.
- It should be grown in a large volume of broth and
then divided into small freezer vials.
- It is used for academic and industrial purposes.
c) Solid medium
- It contains 2-3% agar.
- Some examples include triple sugar iron agar, MacConkey agar (MAC), blood agar plate (BAP), and chocolate agar plate
(CAP)
Notes to Remember:
Agar is a sulfate polymer made of D-galactose-3,6-anhydro-L-galactose and D-glucuronic acid is usually from red algae.
Agar will melt at 80°C to 90°C and solidify at 40 °C to 50 °C .
The cooling temperature for distribution of culture medium into petri dish plate is 55 °C to 60 °C .
CLASSIFICATION OF
CULTURE MEDIA
1. ACCORDING TO CONSISTENCY
a) Liquid Medium
- It does not contain any amount of agar.
- It allows the growth of aerobes, anaerobes, and facultative anaerobes.
- Ex: Brain Heart Infusion, Trypticase soy broth (TSB), Thioglycolate.
b) Semi-solid medium
- It contains 0.5% to 1% agar.
- It is used to observe bacterial motility and to detect indole and sulfide
production.
- Example is Sulfide indole motility (SIM) medium.
2. ACCORDING TO COMPOSITION
a) Synthetic or defined medium
- It is a medium in which all the components are known.
- It is used for research purposes.
- It is preferred for the isolation of cyanobacteria and chemoorganotrophs.
- Example is BG-11 medium.
b) Non-synthetic or complex medium
- It is a medium in which some of the substances are unknowns
- It is used for the isolation of medically significant bacteria.
- Examples: Nutrient broth, MAC, Trypticase soy broth (TSB)
c) Tissue culture medium
- It is used for obligate intracellular bacteria (Rickettsia and Chlamydia)
- Some examples are W138 cells, HeLa 229 cells, and McCoy cells.
- HeLa 229 cells are human cervical tissue cells
- McCoy cells and W138 are fibroblast. It is used for isolation of Chlamydia.
3. ACCORDING TO THE DISPENSING OR DISTRIBUTION METHOD FOR THE MEDIUM

- Plated media are distributed into the dish or plate.

- Tube media are prepared as either liquid, slant, butt and slant, or butt.

4. ACCORDING TO USE

a) Simple media, general purpose media, and supportive media

- These are routinely used in the laboratory and without additional supplements.

- These are the media that support the growth of most non-fastidious bacteria

- Examples: Nutrient agar, Nutrient broth and Trypticase soy broth.

b) Enrichment media (Liquid-type media)

- These are used to propagate the growth of certain group of bacteria from a mixture of organisms.

- These media are incubated for a certain period and then subculture to isolate desired organism.

- Examples: Alkaline Peptone water, selenite F, thioglycolate, Lim broth, Gram-negative broth (GN)
c) Enriched media and non-selective media
- These are media with additional supplements such as blood, vitamins, and yeast extract, which are necessary for the growth
of fastidious organism.
- Example is BAP and CAP
- BAP is used to differentiate the hemolytic patterns of bacteria
- CAP is routinely used for Haemophilus species.
d) Differential media
- It allows the visualization of metabolic differences between groups of bacteria.
- Some examples are MAC, BAP, eosin methylene blue (EMB), Hekton enteric agar (HEA)
- MAC differentiates lactose fermenters (pink colony) from non-lactose ferments (colorless)
e) Selective media
- These are media incorporated with antibiotics, dyes, or chemicals to inhibit the growth of other organisms while promoting
the growth of the desired organism
- Example: HEA, MAC, xylose lysine desoxycholate (XLD), bismuth sulfide agar (BSA), mannitol salt agar (MSA), Thayer-
martin agar (TMA)
◦ Alkaline peptone water used to promote the growth of Vibrio.

◦ Selenite F used to isolate Salmonella from feces, urine, and water samples

◦ Thioglycollate is a general support enrichment medium that promotes the growth of most non- fastidious bacteria.

◦ Tetrathionate is selective enrichment broth for the isolation of Salmonella and Proteus.

◦ Gram- negative broth is used for the isolation of Salmonella and Shigella

◦ Lim broth is used as enrichment medium of Group B Streptococci.

Other selective Media include the following:

1. Gentamicin blood agar for Streptococcus.

2. Bacitracin chocolate agar for Haemophilus.

3. Blood agar plate with ampicillin for Aeromonas.

4. Phenylethyl alcohol for Gram-positive bacteria

5. Columbia colistin-nalidix (CAN) agar for Gram-positive bacteria.

Inhibitory substances that are added to the culture media to isolate the desired organism and make a medium selective.

a) For Gram-positive bacteria: Crystal violet/Gentian violet, basic/carbol fuchsin, and bile salt.

b) For Gram- negative bacteria- Potassium tellurite and sodium azide

c) For swarming bacteria- Alcohol and chloral hydrate.

◦ HEA contains bile salt and dyes (acid fuchsin and ph indicator bromthymol blue) that inhibit the indigenous microbiota of
the lower GIT

◦ MAC contains bile salt and crystal violet that inhibit Gram-positive bacteria.

f) Special media

- LJ medium is a protein rich medium composed of whole eggs and malachite green and which supports the growth of
Mycobacteria. It is sterilized by inspissation.

- TCBS is selective for the isolation of Vibrio in which sterilization is performed by boiling, and never by autoclaving, and
thus considered as a special medium.
 INOCULATION OF SPECIMEN
◦ Sterile body fluids, pus, urine, and sputum, are inoculated directly into the selected media.

◦ Specimens are received on swabs inoculated directly into the culture media.

◦ Some specimens requires direct or bedside inoculation are blood, genital specimens, corneal scrapings, sterile
fluids.
INOCULATION TECHNIQUES
1. STREAKING
- The most common manner of inoculation.
- Overlapping inoculation is used for antimicrobial sensitivity test (disk diffusion method)
MANNER OF INOCULATION
MANNER OF REPORTING (GRADING) OF
GROWTH PLATE
◦ 4+ - Signifies many, heavy growth; if growth is up to the 4th quadrant.
◦ 3+ - Signifies a moderate growth; if growth is up to the 3rd quadrant.
◦ 2+ - Signifies a few or light growth; if growth is in the 2nd quadrant.
◦ 1+ - Signifies a rare growth; if growth is in the 1st quadrant only.
CULTURE MEDIA and THEIR PURPOSES
 ANEROBIC CULTIVATION
WAYS TO FACILITATE ANAEROBIC CULTIVATION
1. The use of a special culture medium incorporated with thioglycollate and cysteine (reducing agents)
2. The boiling point of culture medium to remove (drive off) oxygen.
3. The use of an anaerobic chamber system with a vacuum pump and nitrogen gas to remove residual oxygen.
4. The use of gas-pak jar containing a palladium catalyst.
5. For small-volumes, plastic bags ang pouches containing calcium carbonate and catalyst can be used.

COMPONENTS OF AN ANAEROBIC CHAMBER

1. Nitrogen gas- is used as a filler for the remaining percentage of the anaerobic atmosphere
2. Palladium pellets-are used to remove residual oxygen from the chamber by combining with hydrogen to form water
3. Silica dust(dessicant)- is used to absorb the water that is formed when hydrogen combines with the free oxygen in the
presence of the catalyst
4. Methylene blue or reazurin- is an oxygen reduction indicator that becomes colorless in the absence of oxygen
 ANAEROBIC INCUBATOR
◦ Its components are desiccant, oxygen-free indicator, catalyst and an anerobic source of gas ( Nitrogen,

hydrogen and carbon dioxide)

◦ Anaerobic chamber is the ideal incubation system.

◦ The ideal temperature to promote growth of anaerobes is 37°C for 48 hrs.

 GAS-PAK JAR
◦ For gas-pak jar, it takes 30-45 minutes to obtain an anerobic environment.

◦ When water is added to the jar enveloped, carbon dioxide and hydrogen are produced.

◦ If the catalyst is working accurately, water vapor will be present inside the jar and the indicator strip will be
colorless.

◦ It take several hours for methylene blue indicator to change from blue to colorless

◦ Failure to achieve an anaerobic condition could be the result of a “poisoned” catalyst or a crack in the O-ring
jar or lid.
 STERILIZATION
AND
DISINFECTION
◦STERILIZATION- It refers to the
removal or destruction of all forms of life
including bacterial spores.
 PHYSICAL METHOD OF
STERILIZATION
◦ APPLICATION OF HEAT
Heat is the most common method used to remove or to kill microorganisms.
1. Moist heat procedure
- It destroys microorganisms through coagulation of enzymes and structural proteins and degragation of
nucleic acids.
2 methods of moist heat procedure:
a) Boiling
- It destroys vegetative bacteria when exposed to temperature of 100°C for 10-15 minutes.
b) Autoclaving
- This is the Gold standard of sterilization
- It is the fastest and simplest method of
sterilization through which all organisms
except prions are killed within 15
minutes.
- It is used for sterilization of biohazard
trash and heat-stable objects.
- The principle used is steam under
pressure.
- The biological indicator of autoclave is
Geobacillus stearrothermophilus.
- The method of autoclaving is done with
the use of autoclave which is set into
121°C, 15 psi, 15 minutes for media
liquids, utensils, glass pipettes, and
instruments for assay.
- It is also done in 132 °C, 15psi, 30-60
minutes for decontaminating medical
wastes.
c) Fractional/ Intermittent sterilization (Tyndallization)

- It destroys vegetative cells and spores after 3 consecutive days of sterilization.

- The temperature and time of exposure used are 100°C for 30 minutes using Arnold’s
sterilizer.

d) Inspissation

- It is used to sterilized protein rich medium such as Lowenstein-Jenssen medium.

- Thickening of media through evaporation is the principle used.

- The temperature and time of exposure used are 70 °C to 80 °C for 2 hours; it is performed
for 3 consecutive days.
e) Pasteurization/ Partial sterilization
- It uses to sterilized milk, dairy products, and alcoholic beverages.

- It eliminates food-borne pathogens and organisms responsible for food spoilage.

3 Types of Pasteurization
i. Low-temperature holding/ batch method
- It destroys milk-borne pathogens
- Treatment at this temperature reduces the spoilage of food without affecting its taste.
- Temperature and time requirement is 63 °C for 30 minutes.
ii. High temperature short time/ Flash pasteurization
-72 °C for 15 seconds ( quick heating and immediate cooling)
iii) Ultra-high temperature holding
- 140 °C for 3 seconds (cooled immediately in vacuum chamber)
- Advantage: milk can be stored in room temp for 2 months without affecting its flavor.
2. Dry heat procedure
- It is a sterilization method that does not require water.
- It kills microorganism by denaturating proteins.
- It is utilized in glass wares, oil products, and powder.
- The biological indicator is Bacillus subtilis var. niger
3 methods:
a) Flaming or direct heating.
b) Oven-heating
- It is used for glass wares, oil, petrolatum or powders
- 160 °C for 1.5-2 hours.
c) Incineration
-it is the most common method of treating infectious waste and infected lab animals.
-It’s Principle is burning of materials into ashes at 300 °C to 400 °C
DRYING
OVEN
 MICRO
INCINERATOR
d) Cremation
- It is used to control
the spread of
communicable diseases.
 CHEMICAL METHODS OF
STERILIZATION
◦ DISINFECTION
- Refers to the removal, inhibition, or killing of microorganisms which
include potential pathogens, by using chemical agents usually on
inanimate objects.
 Mechanisms by which chemical disnfectants
destroys microorganisms
1.Reaction with components of the cytoplasmic
membrane.
2.Denaturation of cellular proteins.
3.Reaction with thiol group of enzymes.
4.Damage to DNA and RNA
 Factors that Influence the activity of
disinfectants
1. Types and number of organisms present.

2. Temperature and ph level of the process.

3. Concentration of disinfectants.

4. Amount of organic present.

5. Length of contact time between chemical agent and microorganisms.

6. Type of water used for disinfection.

1. Antiseptic- it is applies topically on the skin. It inhibit sepsis formation.
Example: tincture of iodine, hexachlorophene, phisohex.

2. Disinfectant- it is applied to inanimate objects. Examples are lysol, cresols,

chlorine, sodium hypochlorite 1:10 dilution is an effective disinfectant.

3. Bactericidal- it precipitates bacterial protein, examples are thee H2SO4, HCL.

It kills bacteria in the specimen.

4. Bacteriostatic- it inhibit the growth of bacteria.

 Common Chemical Agents Used in Microbial
Control
1. Acid and alkaline solutions
-it hydrolyzes and coagulates proteins.
2. Phenol
- It is the first widely used antiseptic and disinfectant.
- It destroys plasma membranes and denatures cell proteins.
- 5%n phenol with 10-30 minutes contact time kills Mycobacteria.
3. Halogen
- It destroys microorganisms through the oxidation process.
- Examples are chlorine, iodine, fluorine, bromine, and astatine.
- Tincture of iodine is effective as an antiseptic.
a) IODOPHOR

- It is more stable than iodine.

- Contact time: 30-60 seconds prior onto the skin prior to blood collection.

- Disadvatage: non-sporicidal

b) CHLORINE

- It is used in the form of hypochlorite

- Concentrated solutions should not be used because it is corrosive.

- The contact time is 3 minutes, 10-30 minutes for mycobacteria.

◦ A tincture of iodine contains 2% iodine in 70% alcohol.

◦ Before drawing blood for blood culture, iodophor is applied after 70% alcohol.
4. Alcohol

- It causes the denaturation of proteins and dissolution of lipid membranes.

- It is used as an antiseptic and disinfectant. ( Bactericidal and Fungicidal).

- It should be 60-90% concentration.

- Examples: isopropanol and ethanol.

5. Salt of heavy metal

- It destroys microorganisms by inactivating and precipitating cell proteins.

- Examples: copper, arsenic, mercury, silver, and zinc

- AgNO3 is an eyedrop solution used for Neissseria gonorrhoea while mercuric chloride is an antiseptic.
6. Quaternary ammonium compounds (Quats)

a) Detergent (Non-tuberculocidal and non-sporicidal)

- It is widely used as surface-active agents.

- It disrupts cell membrane.

- It is used on bench-tops, floors, dairy utensils, and retaurants.

- Pseudomonas aeruginosa is resistant to Quats.

b) Phenolics

- These are molecules of phenol that have been substituted by halogens, alkyl, phenyl or benzyl groups.

- They are found in germicidal soaps.

- They are used in hospital floors and surroundings.

7. Aldehydes

- Their antibacterial effect is the inactivation of proteins and nucleic acids.

- They are commonly used in sterilizing medical equipment.

- Examples: 8% formaldehyde and 2% glutaraldehyde.

a) FROMALDEHYDE

- It is commonly used in sterilizing HEPA filters in a biological safety sabinet.

b) GLUTARALDEHYDE

- It is effective against HIV and HBV organisms exposed for 10 minutes.

8. Gas sterilant

a) Ethylene oxide

- It is the most commonly used gas for sterilization. It has explosive property.

- The biological indicator is Bacillus.

- The concentration used from 450mg/L to 700mg/L of chambers space at 55°C to 60 °C .

b) Periacetic acid

- It is active against all vegetative microorganisms and fungal spores.

- It is usually used to sterilize species of medical equipment.

PRINCIPLES OF ANTIMICROBIAL
ACTION AND RESISTANCE
ANTIBIOTICS
These are chemical substances that are
produced by microorganisms with the
capacity to inhibit or to kill other
microorganisms.
SOURCE (MICROORGANISM) ANIBIOTICS PRODUCED
Bacillus subtilis Bacitracin
Bacillus polymyxa Polymyxin
Cephalosporiun Cephalosphorins
Microspora purpurea Gentamicin
Penicillum notatum Penicillin
Streptomyces erythraeus Erythromycin
Streptomyces fradiae Neomycin
Streptomyces nodosus Amphotericin B
Streptomyces noursei Nystatin
Streptomyces venezuelae Chloramphenicol
Streptomyces aureofaciens Tetracycline
Penicillium griseofulvum Griseofulvin
 CLASSIFICATION
OF
ANTIBACTERIAL DRUGS
1. NATURAL DRUGS
- These drugs are produced by bacteria or fungi.
- Examples are: Amphotericin B, erythromycin, kanamycin, neomycin, nystatin, rifampin, streptomycin,
tetracycline, vancomycin…
2. SEMI-SYNTHETIC DRUGS
- These are modified natural drugs with added chemical groups.
- Examples: ampicillin, carbenicillin, methicillin.
3. SYNTHETIC DRUGS
- These are chemically- produced drugs.
- Examples: sulfonamides, trimethoprim, chloramphenicol, ciprofloxacin, isoniazid and dapsone.
 ANTIBIOTIC RESISTANCE
◦ It is a result of both the use and overuse of antimicrobial agents.

◦ It may also arise within antibiotic-producing microorganisms as a mechanism to protect them against the

action of their own antibiotic(autotoxicity).

 INTRINSIC RESISTANCE
◦ I t is a result of the biochemical makeup of wild-type organisms.
◦ It depends on the hydrophobic or hydrophilic nature of the antibiotic and on the impermeability of the cell
wall to the antibiotic.
◦ Tolerance to a particular drug or antimicrobial class.
◦ Natural resistance caused by the following:
a) Lack of affinity of the drug for bacterial target.
b) inaccessibility of the drug to the bacterial cell
c) Extrusion of the drug by chromosomally encoded efflux pumps.
d) innate production of enzymes that inactivate the drug.
 BETA LACTAMASES
◦ These are enzymes that chemically inactivate beta-lactam drugs by disrupting and consequently opening the
beta lactam ring of the molecule.
◦ Beta lactam ring structure inactivated penicilloic acid.
◦ The production of Beta lactamases by some bacteria is a significant mechanism contributing to the resistance
to some beta lactam drugs.
Most clinically important Beta lactamases are as follows:
a) Class A enzymes are found on plasmids
b) Class C enzymes are chromosomally located and inducible by exposure to beta lactams.
 ACQUIRED RESISTANCE
◦ It is present only on certain isolates that are different from the parental strain.
◦ It is usually expressed either as a modification of target sites or enzymatic modification of antibiotics.
 PATHOGENESIS
HOST-MICROBE INTERACTION
Pathogenesis is the development of an infection and disease. Certain virulence

agents with mechanisms of resistance against the host protective factors are

involved in the proliferation of microorganisms and the progress of disease.

 INFECTION
TYPES OF INFECTION ACCORDING TO THE CAUSE

1. Autogenous infection

- It is caused from the microbiota of the host.

2. Iatrogenic infection

- It is an infection that occurs as a result of some medical treatment or procedure.

3. Opportunistic Infection

- It is an infection that affects immunocompromised hosts but not the individuals with normal immune system.

4. Nosocomial infection

- It means “hospital acquired” infection

- It is a type of infection that is acquired at a healthcare facility.

 URINARY TRACT
INFECTION
LUNG INFECTION
4 Common (PNEUMONIA)
types of SURGICAL SITE
INFECTION
Infection
BLOOD STREAM
INFECTION
TYPES OF INFECTION ACCORDING TO HOST DISTRIBUTION
1. Local infection
- It means signs and symptoms are confined in one area.
- Example is infected wound, boil, and abscess.
2. Focal infection
- It starts as a local infection before spreading it to the body.
- Example: tooth infection, tonsilitis, appendicitis and wound infection caused by Clostridium tetani.
3. Systemic infection
- Microbes are be able to spread all over the body, blood and lymphnodes.
4 Types of systemic infection
a) Bacteremia
- Presence of bacteria in the blood.
- It invade bloodstream without active multiplication
- The highest concentration of bacteria is before the fever strikes.
b) Septicemia

- Active multiplication of bacteria inside the blood.

c) Pyemia

- It is a condition wherein pus-producing organisms repeatedly invading the body.

d) Toxemia

- It is the presence of toxins in the blood.

- The toxins are absorbed by the body cells.

 EXTENT OF INFECTION
1. PRIMARY INFECTION
- It is the initial infection that causes the illness. Example: common cold.
2. SECONDARY INFECTION
- It is caused by opportunistic pathogens after the primary infection has weakened the host’s immune system.
- Example: Pneumonia, and bronchitis
3. LARENT INFECTION (SILENT PHASE)
- It is clinically silent inside the body and causes no noticeable illness in the host.
- Example: Asymptomatic polio infection.
4. MIXED INFECTION
- It is caused by two or more organisms.
- Example is wound infection.
5. ACUTE INFECTION
- It is a type of infection that develops and progress slowly.
- Example: whooping cough.
6. CHRONIC INFECTION
- It develops slowly with milder but longer-lasting symptoms.
- Example: Tuberculosis.
 ROUTES OF INFECTION
1. DIRECT TRANSMISSION
a) Congenital- S. agalactiae, N. gonorrhea, T. pallidum
b) Sexual- N. gonorrhea, T. pallidum subsp. Pallidum, C. trachomatis
c) Infectious respiratory secretions- S. pyogenes, N. meningitidis
d) Hand to hand transmission- Rhinovirus
2. INDIRECT TRANSMISSION
a) Fomites
b) Water – Salmonella, Shigella, and Vibrio
c) Arthropod vectors- Borrelia, Francisella, Yersinia
◦ It is a specific illness or disorder that is characterized by a
recognizable set of sign and symptoms which are
attributable to heredity, infection, diet, or environment.
DISEASE
CLASSIFICATION OF
INFECTIOUS DISEASE
1. COMMUNICABLE DISEASE
- It spreads from one host to another.
- Examples: TB, small pox, flu, chicken pox.

2. NON- COMMUNICABLE DISEASE

- It does not spread from one host to another.
- Example: tetanus, botulism.
 CLASSIFICATION OF DISEASE
ACCORDING TO OCCURENCE
1. SPORADIC
- It occurs occasionally
2. ENDEMIC
- It is constantly present in a particular location or population.
3. EPIDEMIC
- It is a disease that affects a large number of people in a given population within a short period of time.
4. PANDEMIC
- It affects population across large regions, several countries or continents or around the world.
 EFFECTS OF INFECTIOUS DISEASE
1. SIGNS
- These are objective changes that can be measured.
- Example: fever, redness, swelling, paralysis
2. SYMPTOMS
- These are the subjective changes, indications of the disease in a person.
- Example: pain, malaise.
3. SYNDROME
- It is a group of signs and symptoms that are associated with a disease.
- Example: AIDS.
 PHASES OF INFECTIOUS DISEASE
1. INCUBATION PERIOD
- Time between the exposure to a pathogenic organism and the onset of symptoms.
2. PRODROMAL PERIOD
- It is the appearance of signs and symptoms.
3. CLINICAL OR ILLNESS PERIOD
- It is the peak of characteristic signs and symptoms of an infection or a disease.
4. DECLINE PERIOD
- Period which the signs and symptoms begin to subside.
5. CONVALESCENCE OR PERIOD OF RECOVERY
- The individual is recuperating towards full recovery.
1. TRUE PATHOGEN
- Organisms that are able to invade the tissues of healthy
individuals through some inherent ability causing various
diseases.
2. OPPORTUNISTIC PATHOGENS
GENERAL CLASSES
- Organisms that normally do not cause disease in their
OF PATHOGENIC
natural habitat to a healthy person. MICROORGANISM
- They cause diseases if the host is immunocompromised
or if they enter a different part of the body.