MCB 221 (Viruses and Sterilization)

Viruses
 Viruses are obligate intracellular parasites.

 Viruses show some of the features of a living organism, such as the ability to
replicate themselves,
 But outside the cell they are just inert chemical
 50x smaller than average bacteria
 Viruses can infect bacteria, algae, fungi, protozoa, invertebrates, vertebrates
and plants.
 Viruses that infect bacteria are called bacteriophages or phages
They differ from true organisms by the following :
1. They cannot be observed using a light microscope.
2. They have no internal cellular structure.
3. They contain either DNA or RNA.
4. They are incapable of replication unless occupying an appropriate living host cell.
5. They are incapable of metabolism 6. Individuals show no increase in size.
Structure
They have a simple structure made up of nucleic acid and protein.
An intact viral particle(virion), has two components: a core of nucleic acid, surrounded
and protected by a protein coat (capsid), the combination of the two being known as the
nucleocapsid In certain virus types, the nucleocapsid is further surrounded by a
membranous envelope, partly derived from host cell material.
Virus Structure
1. Helical Viruses
2. Polyhedral Viruses
3. Complex Viruses
Assignment
Draw a well labelled diagram of a representive virus for the three Viral structure
Sterilization
• Decontamination: process where hazardous substances (Mcos, their toxins and
other chemicals) are removed from the surface or withing an article or substance.
• Sterilization: complete killing of all forms of microorganisms, including bacterial
spores
• Disinfection: killing or removing of harmful vegetative microorganisms.
• Disinfectant: chemical substance used to achieve disinfection.
• Antiseptic: are mild form of disinfectants that can be safely used externally on
living tissues. i.e on the surface of skin and mucous membrane.
• cidal: to kill e.g virucidal, fungicidal, bactericidal, sporicidal etc.
• Stasis: inhibit
Method of Sterilization
1. Physical methods:
• Heat ( dry heat & moist heat)
• Radiation
• Drying
• Filtration
• Low temperature
2. Chemical method: (used for heat sensitive equipments)
• Liquid (Alcohol, Chlorine, Iodine ...)
• Gaseous (Formaldehyde, Glutaraldehyde ...)
Physical Methods
HEAT : Most important should be used whenever possible , types:

1. Dry heat
 Hot air Oven: at temperature of 160˚C for one hour or 180 ˚C for 20
minutes; glassware, powder can be subjected to Oven
 Red heat: wireloops, forceps, spatula are sterilized by holding them in
Bunsen flame till they become red hot.
 Flaming: mouth of testtubes, flask, glass slide, needles, scissors, cover
slip are passes over a bunsen flame (most vegetative cell are killed).
 Incineration: for sterilization of disposable items used in the lab e.g petri
dishes, needles
2. Moist heat
Heating below 100 ˚C:
Pasteurization: process of removing pathogenic substances in substance like milk
(Hold method : milk is heated at 63 ˚C for 30mins; Flash type: heated at 72 ˚C for 15-
20seconds and Ultra : heated at 83 ˚C for 4-5 secs). Suitable to destroy most milk borne
pathogens like Salmonella, Mycobacterium, Streptococcus, Staphylococcus and
Brucella.
Heating at 100 ˚C:
 Boiling; most of the vegetative cell and viruses can be kill immediately at 100 ˚C
for 30 mins.
 Steam at 100 ˚C: media such has DCA, TCBS are sterilized by steaming for 3
consecutive days for 30mins. At the end of the last day of heating, all spores are to be
killed, the process is known as Tyndallisation (after John Tyndall). Sugar and gelating in
the medium may get decomposed on autoclaving.
Heating above 100 ˚C eg. Autoclave
at 121 ˚C for 15mins or 134 ˚C for 4 minutes , for sterilization of liquid media for the
elimination of both the spores and the vegetative cell.
Other physical methods :

Radiation
 Ionizing radiation- (X-rays, Gamma rays..)
– e.g. Gamma radiation: has greater energy than U.V. light, therefore more
effective. Used mainly in industrial facilities e.g. sterilization of disposable
plastic syringes, gloves, specimens containers and Petri dishes.
– leads to the removal of electrons from the cell membrane of the bacteria
cells leading to the formation of ions. The ions react wit protein and
nucleic acid in the bacteria cells leading to their death.
 Non-ionizable radiation (UV and Infrared rays)
– The rays leads to the formation of Thiamer diamer, a process that block
DNA replication and protein synthesis.
– Has limited sterilizing power because of poor penetration into most
materials. Generally used in irradiation of air in certain areas such as
operating rooms and tuberculosis labs.
Filteration
– Filteration does not kill microbes, it separates them out (remove particles from
solution that cant be autoclave).
– May be done under either negative or positive pressure. Example ; membrane
filter made of cellulose acetate with pore size between 0.2-045Կm. Generally
removes most bacteria but viruses and some small bacteria e.g. Chlamydia &
Mycoplasma may pass through. Thus filtration does not technically sterilize
items but is adequate for circumstances under which is used.
– Main use: for heat labile substances e.g. sera, antibiotics.
Drying: Removal of water from the sample , this lead to the death of the
vegetative cells of microorganisms.
Low temperature: Refrigeration and freezing are used to reduce or kill some
vegetative cells of bacteria.
Sterilization by Chemical methods
Useful for heat sensitive materials e.g. plastics and lensed endoscopes).
1. Alcohol: are good disinfecting agent, it remove water from bacterial cell, wash
away the lipid components of lipid polysaccharide layer, it interferes with protein
in the bacteria.
2. Chlorine: its use in municipal water treatment and salt of chlorine Sodium
hypochloride. It interferes with protein synthesis.
3. Formaldehyde (a gas): it can be used to fumigate operation room, surgical wards
etc. It interferes with nucleic acid in bacteria.
4. Ethylene Oxide Chamber
Ethylene oxide alkylates DNA molecules and thereby inactivates microorganisms.
Ethylenes oxide may cause explosion if used pure so it is

mixed with an inert gas .Requires high humidity (50-60% ) .Temperature : 55-60°C
and exposure period 4-6 hours.
5. Activated alkaline Gluteraldehyde 2%
Immerse item in solution for about 20 mins. If Mycobacterium tuberculosis or
spores present then immersion period 2-3 hours. Interfere with nucleic acid synthesis.
Important point
• Any instrument or item used for sterile body site should be sterile.
• Any instrument or item used for non-sterile body site can be disinfected.
• Hand washing is the most important to prevent hospital acquired infection.
Questions
1. What is the differences between:
i. Sterilization and disinfection
ii. Tyndallization and pasteurization
2. Enumerate the methods of moists and heat heat method of sterilization with
examples

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Viruses

 Viruses are obligate intracellular parasites.

HEAT : Most important should be used whenever possible , types:

Other physical methods :

 Non-ionizable radiation (UV and Infrared rays)

Sterilization by Chemical methods

Ethylenes oxide may cause explosion if used pure so it is

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