Basic laboratory glasswares and equipments

Plus sterilization/ disinfection techniques

Sterilization Disinfection

Moist heat (autoclave, tyndalllization, Physical

inspissation)

Dry heat (hot air oven, incineration) Boiling

Pasteurization

Filtration non ionizing radiation(UV rays)

Ionizing radiation(gammarays) chemical

Chemicals Alcohol, aldehydes, halogens, peracetic

acid,H202,quaternary ammoniums,phenols
(ethylene oxide, peracetic acids,etc)
Factors That Influence the Degree of Killing Before discussing methods used to kill microorganisms, a
review of the factors that influence the degree of killing of organisms is important. The following factors
play a significant role in the selection and implementation of the appropriate method of disinfection:

• Types of organisms
• Number of organisms
• Concentration of disinfecting agent
• Presence of organic material (e.g., serum, blood)
• Nature of surface to be disinfected
• Contact time
• Temperature
• pH
• Biofilms
• Compatibility of disinfectants and sterilants
Parts:

 pressure chamber
 pipes to introduce
 evacuate steam, valves to remove air and control pressure
 pressure and temperature gauges to monitor the procedure
MOA: As steam enters an autoclave chamber, it forces air out, raises the temperature of the contents,
and increases the pressure until a set temperature and pressure are reached.

* The relationship between temperature and pressure. Note that higher temperatures—and
consequently, greater antimicrobial action—are associated with higher pressures

I. Alcohols
A. are ethyl alcohol and isopropyl alcohol. Alcohols have excellent in vitro bactericidal
activity against most gram-positive and gram-negative bacteria.
B. They also kill Mycobacterium tuberculosis various fungi, and inactivate certain
enveloped viruses; however, they are not sporicidal and have poor activity against
nonenveloped viruses
II. Aldehydes
1. Formaldehyde
a. Formaldehyde is generally used as formalin, a 37% aqueous solution, or
formaldehyde gas.
b. Formaldehyde gas is often used to disinfectant biosafety hoods.
c. its usefulness is limited by its irritability factor and its potential carcinogenicity
 2. Glutaraldehyde
a. is a saturated five-carbon dialdehyde that has broadspectrum activity and rapid killing
action and remains active in the presence of organic matter.
b. Glutaraldehyde is extremely susceptible to pH changes and is active only in an alkaline
environment.
c. When used as a 2% solution, it is germicidal in approximately 10 minutes and sporicidal
in 3 to 10 hours. Its killing activity is due to inactivation of DNA and RNA through
alkylation of sulfhydryl and amino groups
d. is bactericidal, pseudomonacidal, fungicidal, and virucidal (against human
immunodeficiency virus [HIV] and hepatitis B virus [HBV]) with a minimum of 10
minutes’ exposure at a temperature between 20° and 30°C. It is also tuberculocidal.
III. Halogens
1. Iodophors
a. Iodine can be used as a disinfectant in one of two forms: tincture or iodophor.
b. Tinctures are alcohol and iodine solutions, used mainly as antiseptics.
c. An iodophor is a combination of iodine and a neutral polymer carrier that
increases the solubility of the agent. This combination allows the slow release of
iodine. Iodophors must be diluted properly for them to be effective.
d. Iodophors have the added advantage of being less irritating, nonstaining, and
more stable than iodine in its pure form. Iodophors may be used as antiseptics
or disinfectants, depending on the concentration of free iodine.
e. The best known iodophor is povidone-iodine (Betadine), which is mainly used
as an antiseptic. Povidone-iodine provides slow and continuous release of free
iodine.
f. Free iodine degrades microbial cell walls and cytoplasm, denatures enzymes,
and coagulates chromosomal material
IV. Chlorine and Chlorine Compounds
a. are usually used in the form of hypochlorite, such as the liquid sodium hypochlorite
(household bleach) and solid calcium hypochlorite

b.Their killing activity is based on the oxidative effects of hypochlorous acid, formed when
chloride ions are dissolved in water

c. The most common use of chlorine is disinfection of water.

V. Detergents: Quaternary Ammonium Compounds
a. Substitution of the four-valence ammonium ion with alkyl halides.
 b. They are cationic, surface-active agents, or surfactants, that work by reducing the
surface tension of molecules in a liquid

phenol coefficient :
A. expression of bactericidal power if a substance as compared to a pure phenol
B. bpc > or /greater than 1 : disinfectant is better than phenol
C. PC < or / less than 1: phenol is better
D. Pc = 1: same efficiency

biosafety cabinet
 a. The class I biological safety cabinet (hood) uses an exhaust fan to move air inward through the
open front. The air is circulated within the safety hood, passing through a high efficiency
particulate air (HEPA) filter before reaching the environment outside the hood
b. The class II biological safety hood is the most common in microbiology laboratories. Air is
pulled inward and downward by a blower and passed up through the air flow plenum, where it
passes through a HEPA filter before reaching the work surface. A percentage of the remaining
air is HEPA filtered before reaching the environment.
c. The class III biological safety hood is a selfcontained ventilated system for highly infectious
microorganisms or materials and provides the highest level of personal protection. The closed
front contains attached gloves for manipulation on the work surface

Biosafety Levels
1 : agents include those that have no known potential for infecting healthy people and are well defined
and characterized. BSL-1 agents include Bacillus subtilis and Naegleria gruberi;

2: agents are those most commonly being sought in clinical specimens and used in diagnostic, teaching,
and other laboratories. They include all the common agents of infectious disease, as well as HIV,
hepatitis B virus, Salmonella organisms, and several more unusual pathogens.

3: procedures have been recommended for the handling of material suspected of harboring organisms
unlikely to be encountered in a routine clinical laboratory and for such organisms as Mycobacterium
tuberculosis, Coxiella burnetii, and the mold stages of systemic fungi and for some other organisms
when grown in quantities greater than that found in patient specimens

4: agents are exotic agents that are considered high risk and cause life-threatening disease. They
include Marburg virus or Congo-Crimean hemorrhagic fever.