Bacterial

Staining
WILSON R. DELOS REYES JR.,RMT, MLS (ASCPi), MSMT, MD
Kinds of Ionizable Dyes Used in Staining
Bacteria
BASIC DYES
- Commonly used
- Cationic dyes with positively charged groups that adhere to
negatively charge molecules like nucleic acids and proteins.
- Example: Methylene blue, crystal violet, safranin and malachite
green
Kinds of Ionizable Dyes Used in Staining
Bacteria
ACIDIC DYES
- Anionic dyes with negatively charged groups that bind to
positively charge cell structures.
- Example: Eosin and acid fuchsin
Staining Techniques
SIMPLE STAINING
- Single stain is used
- Directed towards coloring the forms and shape of the cells
- Example: Methylene blue
Staining Techniques
DIFFERENTIAL STAINING
- Divide bacteria into separate groups
- Directed towards coloring the components of the elements
present.
- Example: Gram staining and Acid-fast bacilli (AFB) staining
Staining Techniques
DIFFERENTIAL STAINING
- Steps in differential staining are as follows:
- 1. Application of Primary stain
- 2. Application of the mordant
- 3. Application of the decolorizing agent
- 4. Application of the secondary stain/counterstain
Staining Techniques
NEGATIVE STAINING
- Demonstrate presence of diffuse capsule surrounding some
bacteria
- Excellent technique for studying bacterial gas vacuole and viral
morphology
- Appearance: bacteria as light-colored bodies against dark
background
- Example: India Ink or Nigrosin dye
Gram Stain
- Most commonly used differential stain
- Utilizes crystal violet as the primary stain, while safranin is the
secondary stain or counterstain.
- Iodine - act as the mordant
- Acetone alcohol - act as decolorizing agent
 GRAM-POSITIVE BACTERIA GRAM-NEGATIVE BACTERIA

PRIMARY STAIN
(Crystal violet)

MORDANT
(Iodine)

DECOLORIZATION
(Acetone)

SECONDARY STAIN
(Safranin)
General Rule of Gram staining
- All cocci are Gram positive
except:

- Neisseria

- Veilonella

- Branhamella (Moraxella)
General Rule of Gram staining
- All bacilli are Gram
negative except:

- Arcanobacterium - Listeria
- Bacillus - Mycobacterium
- Clostridium - Nocardia
- Corynebacterium - Streptomyces
- Erysipelothrix - Trophyrema whipplei
Reason why Gram-positive become Gram-negative
bacteria
- Removal of MgRNA
- Aged, dying and autolyzing cells
- Old cells may lose their ability to retain strains
- Antibiotic-treated bacterial cells have atypical staining
reaction
- Using acidic iodine during staining
- Due to a technical error or the wrong use of stains
Exception in Gram staining

- Organisms that exists almost exclusively within host cells

(Chlamydia)
- Organisms that lack cell walls (Mycoplasma and Ureaplasma)
- Organisms with insufficient dimension to be resolved by light
microscopy (Spirochetes).
Acid-Fast Staining

- Used to stain bacteria that have high lipid contents in their cell
wall
- Utilizes carbol fuchsin as the primary stain and methylene blue or
malachite green as the secondary stain.
- Cell wall of acid fast bacteria resists the acid-alcohol in
decolorizing step.
- Heat - applied as a mordant in Ziehl-Neelsen method
- Tergitol - applied in Kinyoun method.
 ACID-FAST BACTERIA NON-ACID FAST BACTERIA

PRIMARY STAIN
(Carbol fuchsin)

MORDANT
(Heat/Tergitol)

DECOLORIZATION
(3% acid-alcohol)

SECONDARY STAIN
(Methylene blue)
Acid-Fast Staining Method

- 1. Ziehl-Neelsen/Hot staining Method

- 2. Kinyoun’s/Cold staining Method
- 3. Pappenheim method - differentiate Mycobacteriuam smegmatis
from Mycobacterium tuberculosis
- 4. Baumgarten method - differentiate Mycobacterium leprae from
Mycobacterium tuberculosis.
- 5. Auramine-rhodamine method - selective for the cell wall of
AFB.
Modified Acid-Fast Staining Method (Modified Kinyoun)

- Useful for the identification of intestinal coccidian oocysts

- Ideal for cryptosporidia and cyclospora parasites
- Specimen : stool
- Reagent: same with conventional acid-fast except the
concentration of acid-alcohol (1% H2SO4)
- Result: oocysts appears as magenta-stained organisms against a
blue background
Special Staining Method

Staining Technique Cellular structure/Bacteria

Dyar Cell Wall
Anthony’s, Hiss and Gin’s Capsule
Nigrosin Capsule
Neisser Metachromatic granules
Albert Metachromatic granules
Dorner Endospore
Special Staining Method

Staining Technique Cellular structure/Bacteria

Schaeffer-Fulton Endospore
Gray Flagella
Leifson Flagella
Feulgen DNA
Levaditi’s Sphirochetes
Fontana-Tribondeau Sphirochetes
 CULTURE AND
CULTURE MEDIA
WILSON R. DELOS REYES JR., RMT, MLS(ASCPi), MSMT, MD
CULTURES
- Growth of microorganisms in a culture media. Utilizing effective and
appropriate culture media for growth, transport, and storage facilitates
the study in Bacteriology
CULTURE MEDIA
- Composed of a mixture of nutrients such as carbon, nitrogen, sulfur,
phosphorus, hydrogen, oxygen and buffers
TYPES:
- Liquid, Semi-solid and Solid Medium
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 several culture species contained in a separate culture
medium (one species per culture medium).
- It is used for academic and industrial purposes.
CLASSIFICATION OF CULTURE MEDIA
1. According to Consistency
2. According to Composition
3. According to Dispensing or Distribution Method for the
Medium
4. According to Use
A.ACCORDING TO CONSISTENCY
1. 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) and thioglycollate.
ACCORDING TO CONSISTENCY
2. SEMI-SOLID MEDIUM
- It contains 0.5% to 1% Agar
- It is used to observe bacterial motility and
detect indole and sulfide production
- Ex: Sulfide Indole Motility (SIM) Medium
ACCORDING TO CONSISTENCY
3. SOLID MEDIUM
- It contains 2% to 3% agar
- Ex: Triple sugar Iron (TSI) agar,
MacCOnkey (MAC) agar, Blood agar
plate (BAP) and Chocolate agar plate
(CAP)
B. ACCORDING TO COMPOSITION
1. SYNTHETIC OR DEFINED
MEDIUM
- Medium in which all the
components are known
- Used for research purposes
- Preferred for isolation of
cyanobacterium and
chemoorganotrophs
- Ex: BG-11 medium
B. ACCORDING TO COMPOSITION
2. NON-SYNTHETIC or COMPLEX MEDIUM
- Medium in which some substances are unknown
- Peptones, Meat and Yeast Extract
- Useful for isolation of medically significant bacteria
- Ex: Nutrient broth (NB) broth medium, TSB and MAC agar
B. ACCORDING TO COMPOSITION
3. TISSUE CULTURE MEDIUM
- Used for obligate intracellular bacteria (Rickettsia and
Chlamydia).
- Example: W138 cells, HeLA 299 Cells and McCoy cells
HeLa 299 cells Human cervical tissue
McCoy cells Fibroblast
W138 cells Fibroblast
B. ACCORDING TO COMPOSITION
3. TISSUE CULTURE MEDIUM
C. ACCORDING TO THE DISPENSING OR
DISTRIBUTION
PLATE MEDIA
- Distributed into the dish or plate
TUBE MEDIA
- Prepared as either liquid, slant, butt and slant or butt
EXAMPLES:
- Triple sugar iron (TSI)
- SIM
- Simmon’s citrate agar
- Lysine iron agar (LIA)
C. ACCORDING TO THE
DISPENSING OR
DISTRIBUTION
D. ACCORDING TO USE
1. SIMPLE MEDIA, GENERAL PURPOSE MEDIA AND SUPPORTIVE
MEDIA
- Routinely used in the laboratory and without additional supplement
- Support the growth of most non-fastidious bacteria
- Usually composed of meat and soybean extract
- Ex: Nutrient agar, Nutrient broth and Tryptone soy broth (TSB)
D. ACCORDING TO USE
2. Enrichment media (Liquid-type media)

- Used to propagate the growth of certain group of bacteria from a

mixture of organisms.
- Contain specific nutrients and without additional supplement
- Example: Alkaline peptone water, Selenite F, Thioglycollate,
Tetrathionate, Gram-negative (GN) broth and Lim broth
D. ACCORDING TO USE
3. Enriched media and Non-selective media

- Media with additional supplement such as blood, vitamins and

yeast extract
- Solid-type of media
- Example: Blood agar plate and Chocolate agar plate
D. ACCORDING TO USE
BLOOD AGAR PLATE CHOCOLATE AGAR PLATE
- Hemolytic pattern of bacteria - Recovery of Haemophilus
D. ACCORDING TO USE
4. Differential Media

- These media allow the visualization of metabolic differences

between groups of bacteria
- Example: MAC, BAP, eosin methylene blue (EMB) and Hektoen
Enteric Agar (HEA)
D. ACCORDING TO USE
MacConkey Agar Blood Agar Plate

- Indicator: Neutral red - Differentiate hemolytic pattern of

streptococci
D. ACCORDING TO USE
5. Selective Media

- These media are incorporated with antibiotics, dyes or chemicals

to inhibit the growth of other organisms.
- Example: HEA, MAC, Xylose lysine desoxycholate (XLD) agar,
Bismuth sulfate agar (BSA), Mannitol Salt Agar (MSA) and Thayer-
Martin Agar (TMA)
D. ACCORDING TO USE
5. Selective Media

Other selective media:

- Gentamicin blood agar: Streptococcus
- Bacitracin chocolate agar: Haemophilus
- Blood agar plate with ampicillin: Aeromonas
- Phenylethyl alcohol: Gram positive bacteria
- Colistin-Nalidixic acid (CNA) agar: Gram positive bacteria
D. ACCORDING TO USE
5. Selective Media

Inhibitory substances

Gram Positive Bacteria Crystal/Gentian violet,

basic/carbol fuchsin and bile salt

Gram Negative Bacteria Potassium tellurite and sodium

azide

For Swarming Bacteria Alcohol and chloral hydrate

D. ACCORDING TO USE
5. Selective Media
MEDIA DESCRIPTION

Hektoen Enteric Agar (HEA) Bile salt and dyes: Inhibit indigenous microbiota of LGIT;
used for recovery of fecal bacteria
pH indicator: Bromthymol blue

MacConkey Agar (MAC) Bile salts and crystal violet: inhibit gram-positive bacteria;
used for recovery of fecal bacteria

Xylose Lysine Desoxycholate Agar (XLD) Xylose, lysine, sucrose, 0.25% sodium desoxycholate
and sodium thiosulfate; for fecal bacteria
Differentiate: Shigella and Salmonella

Mannitol Salt Agar (MSA) Support growth of Staphylococcus aureus

Thayer-Martin Agar (TMA) Selective for Neisseria sp.

Xylose lysine desoxycholate Agar
D. ACCORDING TO USE
6. Special Media

- Used to isolate bacteria with specific growth requirements

- Example: Lowenstein-Jensen (LJ) medium and Thiosulfate citrate–
bile salts-sucrose (TCBS) agar.
D. ACCORDING TO USE
LOWENSTEIN-JENSEN MEDIUM

Composed of Whole eggs and malachite green

STERILIZATION AND DISINFECTION
WILSON R. DELOS REYES JR., RMT, MLS (ASCPi), MSMT, MD
01 STERILIZATION

Refers to the removal or

destruction of all forms
of life, including bacterial
spores
 PHYSICAL
METHOD
1. Application of Heat
2. Filtration
3. Low/Cold Temperature
4. Dessication and
Lyophilization
5. Osmotic Pressure
6. Radiation
Application of Heat
MOIST HEAT
Destroys microorganisms
through coagulation of
enzymes and structural
proteins and degradation
of Nucleic acids
 A. BOILING

Destroys vegetative bacteria (non-sporulating).

TEMP: 100C.

TIME: 10 to 15 minutes.

Content Here Content Here Content Here Content Here Content Here
 B. AUTOCLAVING

Fastest and simplest, all organisms (except for

prions) including those that contains spores
PRINCIPLE: Steam under pressure.

BIOLOGICAL INDICATOR:
- Bacillus stearothermophilus
- New Name: Geobacillus
stearothermophilus.
- 121C, 15 psi for 15 mins (media, liquids,
pipettes, utensils, etc.
- 132C, 15 psi for 30-60 mins decontaminating
medical wastes.
 C. TYNDALLIZATION

Destroys vegetative cells and spores after 3

consecutive days of sterilization
Fractional or Intermittent Sterilization.

TEMP: 100C
TIME: 30 minutes
For 3 consecutive days
Arnold’s sterilizer (free-flowing steam)
 D. INSPISSATION

Sterilize protein-rich medium

Principle: thickening of media through evaporation

TEMP: 70C to 80C

TIME: 2 hours
For 3 consecutive days
Ex. Lowenstein-Jensen medium
 E. PASTEURIZATION

Partial Sterilization

Sterilize milk, dairy products and alcohol

Eliminates food borne pathogens and

organisms responsible for spoilage.

Cannot eliminate bacterial endospores

TYPES OF PASTEURIZATION
ULTRA-HIGH
LOW-TEMPERATURE HIGH TEMPERATURE
TEMPERATURE
HOLDING (LTH) SHORT-TIME (HTST)
(UHT)
BATCH METHOD FLASH PASTEURIZATION --
Milk can be stored at
Destroy milk-borne Destroy milk-borne room temperature for
pathogens pathogens two months without
affecting its flavor

Reduces spoilage of Process also

Quick heating then
food without affecting applicable to the
immediate cooling
taste coffee creamer

63C for 30 minutes 72C for 15 seconds 140C for 3 seconds

Application of Heat
DRY HEAT
- Sterilization method does not require
water
- Kills microorganisms by denaturating
proteins
- Biological indicator: Bacillus subtilis var.
niger
- New name:Bacillus atrophaeus
 A. FLAMING

DIRECT HEATING

Content Here Content Here Content Here Content Here Content Here
 B. OVEN HEATING

For glassware, oil or powders

TEMP: 160C to 170C

TIME: 1.5 to 2 hours

Content Here Content Here Content Here Content Here Content Here
 C. INCINERATION

Most common method of treating infectious

waste and infected animals

Hazardous material:
TEMP: 870C to 980C

Principle: Burning materials into ashes at

300C to 400C

Content Here Content Here Content Here Content Here Content Here
 D. CREMATION

Used to control spread of communicable

disease

Content Here Content Here Content Here Content Here Content Here
 FILTRATION
- Method of choice for sterilization of
antibiotic solution, toxic chemicals,
radioisotopes, vaccines and
carbohydrates
- Both for liquid and air substance
 A. DEPTH FILTERS

Made up of fibrous or granular materials

Ex. Berkefield filter, Chamberland filter and

asbestos

Content Here Content Here Content Here Content Here Content Here
 A. MEMBRANE FILTERS

Porous membranes (0.1 um thick)

Composed of cellulose acetate or

polycarbonate

Used to sterilize pharmaceuticals, ophthalmic

solution, culture media, antibiotics and oil
products

Content Here Content Here Content Here Content Here Content Here
 MEMBRANE FILTERS
LIQUID AIR FILTRATION
FILTRATION OF CRITICAL
FILTRATION BACTERIA, YEAST AND
STERILIZING
MOLDS

Uses cellulose Uses high-efficiency Uses 0.22um

particulate air (HEPA) Uses 0.45um pores
acetate or cellulose membrane filters for
filter of membrane filter
nitrate membrane parenteral solutions
Removes organisms Range: 0.2-0.45um
with a vaccum. and alcohol.
larger than 0.3um Remove most
Removes vegetation
bacteria and fungi
cells and spores but
Text Content Here but not virus
Text Content Here not virus

Text Content Here

Text Content Here
LOW/COLD TEMPERATURE
- Considered bacteriostatic
- Reduces the rate of metabolism
- Important in food microbiology
- 2C to 8C for 72 hours – kills syphilis
DESSICATION and LYOPHILIZATION
Dessication
- Destroys bacteria through disruption of
metabolism involves removing of water from
microbes (bacteriostatic).
Lyophilization
- Destroys bacteria through changes in
proteins and decrease in chemical
reaction
DESSICATION and LYOPHILIZATION
 OSMOTIC PRESSURE
- High concentration of salts and sugars to
create a hypertonic environment
 RADIATION
- PRINCIPLES: Radiation passes through
the cells free hydrogen and hydroxyl
radical and some peroxidase are created.
- Free radicals – cause intracellular
damage
 A. IONIZING RADIATION

Cold Sterilization
Causes mutation in the DNA and produce
peroxidase
Destroy vegetative cells and endospores of
both prokaryotes and eukaryotes
Gamma rays (1500 to 2500 radiation) & xrays

Biological indicator: Bacillus pumilus

Content Here Content Here Content Here Content Here Content Here
 B. NON-IONIZING RADIATION

Damage to cellular DNA by producing thymine dimers

Used on exposed surface, operating rooms

Microorganisms in water are destroyed under

UV lamps
Ultraviolet rays (10um to 400um) in which
260um is the most lethal

Content Here Content Here Content Here Content Here Content Here
CHEMICAL
METHOD
1. Acid and Alkaline solution
2. Phenol
3. Alcohol
4. Halogen
5. Salt of Heavy Metals
6. Quaternary ammonium
7. Aldehydes
8. Gas sterilants
0
DISINFECTION
2
Refers to the removal,
inhibition or killing of
microorganisms usually
on inanimate objects.
Does not remove
bacterial spores.
 Terminology
ANTISEPTIC
0 Content Applied topically on the skin
1 Here Inhibit sepsis formation

DISINFECTANT
0 Content
Applied to inanimate objects
2 Here Lysol, Chlorine and Sodium hypochlorite (1:10).
BACTERICIDAL
0 Content
Precipitates bacterial protein and Kills all
3 Here bacteria in the specimen
Ex. Strong acids.

0 Content BACTERIOSTATIC
4 Here Inhibits the growth of organisms
A. ACID AND ALKALINE SOLUTION

Hydrolyzes and coagulates proteins

Content Here Content Here Content Here Content Here Content Here
 B. PHENOL

Firs widely used antiseptic and disinfectant

Destroys plasma membrane and denatures

protein.
5% Phenol; 10 to 30 mins contact time
Effective against mycobacteria

Ex. Xylenols, lysols, cresols

 C. ALCOHOL

Non-sporicidal, denatures protein and dissolution

of lipid membrane
Both an antiseptic and disinfectant

60% to 90% concentration

Should be allowed to evaporate to achieve
complete antisepsis
Ex. Isopropanol (rubbing alcohol) and ethanol
 D. HALOGEN

Destroys microorganisms through oxidation

Ex. Chlorine, iodine, fluorine and bromine

Tincture of iodine and iodophor are effective

antisepsis
A 1:10 dilution of sodium hypochlorite is an
effective bleach
IODOPHOR HALOGEN 3 minutes
(mycobacteria 10 to 30
mins)
More stable than iodine in its pure
form.
CHLORINE
Combination of iodine and
detergent.
Used in the form of hypochlorite
Antibacterial effect is the oxidative
effect of molecular iodine and
hypoiodic acid CDC recommends a 1:10 dilution of
5.25% solution of sodium
Improperly diluted iodophors may hypochlorite.
not kill microorganisms. Antibacterial activity through oxidative
and disinfecting effects of hypochlorous
30 to 60 seconds acids
onto the skin
Concentrated solution should not
be used for disinfection.
 E. SALTS OF HEAVY METALS

Destroys microorganisms by inactivating and

precipitating cell proteins.
Ex. Copper, arsenic, mercury, silver and zinc

AgNO3 eye drop for Neisseria gonorrhoeae

Mercuric chloride as antiseptic

 F. QUATERNARY AMMONIUM
COMPOUNDS
DETERGENTS
Non-tuberculocidal and Non sporicidal

Most widely used surface active agents

Disruption of the cell membrane leads to

leakage of cell content
Ex. Zephiran (benzalkonium chloride and ceepryn)
And cetylpyridinium chloride

Pseudomonas aeroginosa
in ammonium citrate
medium is resistant to
quats
 F. QUATERNARY AMMONIUM
COMPOUNDS
PHENOLICS
Non sporicidal
Molecules of phenols that have been
substituted by halogens, alkyl, phenyl or
benzyl
Found in germicidal soaps
Use in hospital floors
Antibactericidal effect is cell wall disruption
 G. ALDEHYDES
FORMALDEHYDE (HCHO)
Generally as formalin consist of 37% aqueous
solution
For mycobacteria, 3% to 8% HCHC. Contact
time 30 minutes
Commonly used in sterilizing HEPA filters

Irritability factor and known carcinogen

 G. ALDEHYDES
GLUTARALDEHYDE
Effective against HIV and Hepatitis B for 10 mins

For medical instruments (heat-labile) that are

made of plastics and rubber material
2% glutaraldehyde is germicidal in 10 minutes
Sporicidal in 3 to 10 hours
Rapid killing action but does not penetrate
organic material well.

Pseudomonacidal,
tuberculocidal, fungicidal and
virucidal
 H. GAS STERILANT
ETHYLENE OXIDE (EtO)
Most commonly used gas for sterilization

Antibacterial effect is the alkylation of nucleic

acids in spores and vegetative cells
Concentration: 450 mg/L to 700 mg/L of
chamber space at 55C to 60C for two hours
Used to sterilize plastic petri dishes, sutures,
catheters and heart-lung machines.

Biological indicator: Bacillus

globigii
 H. GAS STERILANT
PERIACETIC ACID
Active against all vegetative microorganisms and
fungal spores

Sterilize pieces of medical equipment

BIOSAFETY IN MICROBIOLOGY
LABORATORY

WILSON R. DELOS REYES JR., RMT, MLS (ASCPi), MSMT, MD

BIOLOGICAL SAFETY CABINET

- Device that enclose a working area to protect workers from

aerosol exposure and infectious disease agents.
- Air that contains the infectious material is sterilized thru:
- Heat
- UV light
- Passage through a high-efficiency particulate (HEPA) resistance
filter
BIOLOGICAL SAFETY CABINET

CLASS I CABINET
- Open-fronted cabinet with negative pressure
- Room air —---- > sterilized using HEPA filter
- Only air to be exhausted is sterlizied
- Used for biosafety levels (BSL) 2 and 3 agents
BIOLOGICAL SAFETY CABINET

CLASS II CABINET
- Also known as Laminar flow BSC
- Most commonly used BSC
- Sterilized air using HEPA filter flows over the infectious
material and the air to be exhausted
- Used for biosafety levels (BSL) 2 and 3 agents
BIOLOGICAL SAFETY CABINET

CLASS II CABINET
- 2 types of Class II cabinet
- A. Class IIA - has fixed opening; 70% of the air recirculated
- B. Class IIB - used for chemicals, radioisotopes and
carcinogens.
* Most hospital clinical microbiology laboratory technologist use
Class II BSC
BIOLOGICAL SAFETY CABINET

CLASS III CABINET

- Provides the highest level of safety to the worker
- Air coming into and going out of the cabinet is sterilized
using HEPA filter.
- Infectious material within is handled with rubber gloves
- Used for biosafety levels (BSL) 4 agents
CLASSIFICATION OF BIOLOGIC AGENTS

BIOSAFETY LEVEL 1 AGENTS

- No known potential for infecting healthy people
- For academic purposes
- Ex: Bacillus subtilis and Naegleria gruberi
CLASSIFICATION OF BIOLOGIC AGENTS

BIOSAFETY LEVEL 2 AGENTS

- Agents acquired by ingestion and exposure to percutaneous
and mucous membrane
- All common agents of infectious diseases
- Ex: HIV, Bacillus anthracis, Yersinia pestis, Salmonella sp.
and Shigella sp.
CLASSIFICATION OF BIOLOGIC AGENTS

BIOSAFETY LEVEL 2 AGENTS

- Agents acquired by ingestion and exposure to percutaneous
and mucous membrane
- All common agents of infectious diseases
- Ex: HIV, Bacillus anthracis, Yersinia pestis, Salmonella sp.
and Shigella sp.
CLASSIFICATION OF BIOLOGIC AGENTS

BIOSAFETY LEVEL 3 AGENTS

- Potential agent for aerosol transmission
- Must have extra precaution in processing lethal pathogen
- Ex: Mycobacterium tuberculosis, Francisella tularensis,
Brucella spp., Coxiella burnetti, St. Louis encephalitis and
systemic fungi
CLASSIFICATION OF BIOLOGIC AGENTS

BIOSAFETY LEVEL 4 AGENTS

- Life-threatening infections
- Needs maximum containment and decontamination of all
personnel and materials before leaving the area.
- Example: arbovirus, arenavirus, filovirus and smallpox virus
CATEGORIES OF POTENTIAL INFECTIOUS AGENT
OF BIOTERRORISM
CATEGORY A
- Agent that poses the greatest public health threat
- Easily transmitted and highly infectious
- Examples: smallpox, Bacillus anthracis and Francisella
tularensis
CATEGORIES OF POTENTIAL INFECTIOUS AGENT
OF BIOTERRORISM
CATEGORY B
- Moderate morbidity and low mortality
- Not easily transmitted
- Examples: Coxiella burnetti, Burkholderia pseudomallei and
Rickettsia sp.
CATEGORIES OF POTENTIAL INFECTIOUS AGENT
OF BIOTERRORISM
CATEGORY C
- Emerging pathogens
- Examples: viruses that causes yellow fever and dengue
hemorrhagic fever.
D. According to Use