Lab 1 Virology Dr.

Nadia Al-Healy
Laboratory Safety
Many viruses are human pathogens, and laboratory-acquired infections
can occur. Laboratory procedures are often potentially hazardous if
proper technique is not followed. Among the common hazards that might
expose laboratory personnel to the risk of infection are the following:
(1) aerosols generated by homogenization of infected tissues,
centrifugation, ultrasonic vibration, or broken glassware.
(2) ingestion from mouth pipetting, eating or smoking in the laboratory,
or inadequate washing of hands
(3) skin penetration from needle sticks, broken glassware, hand
contamination by leaking containers, handling of infected tissues, or
animal bites.
(4) splashes into the eye or mucous membranes.
Good biosafety practices include the following:
(1) training in and use of aseptic techniques.
(2) no eating, drinking, mouth pipetting, or smoking in the laboratory.
(3) use of personal protective equipment (e.g., coats, gloves, or masks)
not to be worn outside the laboratory.
(4) sterilization of experimental wastes.
(5) use of biosafety hoods.
(6) immunization if relevant vaccines are available.
Additional precautions and special containment facilities (Biosafety
Level 4) are necessary when personnel are performing research with
high-risk agents such as the filoviruses and rabies virus.
Identification of a Particle as a Virus
When a characteristic physical particle has been obtained, it should fulfill
the following criteria before it is identified as a virus particle:
1. The particle can be obtained only from infected cells or tissues.
2. Particles obtained from various sources are identical regardless of the
cellular origin in which the virus is grown.
3. Particles contain nucleic acid (DNA or RNA), the sequence of which
is not the same as the species of host cells from which the particles were
obtained.
4. The degree of infective activity of the preparation varies directly with
the number of particles present.
5. Destruction of the physical particle by chemical or physical means is
associated with a loss of viral activity.
6. Certain properties of the particles and infectivity must be shown to be
identical (e.g., their sedimentation behavior in the ultracentrifuge and
their pH stability curves).
7. Antisera prepared against the infectious virus should react with the
characteristic particle and vice versa. Direct observation of an unknown
virus can be accomplished by electron microscopic examination of
aggregate formation in a mixture of antisera and crude viral suspension.
8. The particles should be able to induce the characteristic disease in vivo
(if such experiments are feasible).
9. Passage of the particles in tissue culture should result in the production
of progeny with biologic and antigenic properties of the virus.

Biological hazards, also known as biohazards, refer to biological

substances that pose a threat to the health of living organisms, primarily
that of humans. This can include medical waste or samples of a
microorganism, virus or toxin (from a biological source) that can affect
human health. It can also include substances harmful to animals.
Biological Containment and Laboratory Biosafety Levels:
Laboratories are classified according to their containment potential, or
biosafety level (BSL), and are designated as BSL-1, BSL-2, BSL-3, or
BSL-4. sometimes given as P1 to P4 (for Pathogen or Protection level),
Higher numbers indicate a greater risk to the external environment:
Biosafety level 1:
BSL-1 laboratories are the lowest level of containment. Work can be
done on the open bench with organisms that present a low risk of
infection; these organisms are not pathogens in normal individuals and
include organisms such as Bacillus subtilis. Workers should be protected
by barrier protection such as lab coats and gloves. An example of a BSL-
1 facility is a teaching laboratory that does not use pathogens.
Decontamination procedures for this level are similar in most respects to
modern precautions against everyday microorganisms (i.e., washing one's
hands with anti-bacterial soap, washing all exposed surfaces of the lab
with disinfectants, etc.).
Biosafety level 2
Entities this level is similar to Biosafety Level 1 and is suitable for work
involving agents of moderate potential hazard to personnel and the
environment. It includes various bacteria(Escherichia coli or
Streptococcus pyogenes) and viruses(influenza A, Lyme disease,
mumps, measles) that cause only mild disease to humans, or are difficult
to contract via aerosol in a lab. Normal procedures may be performed on
bench tops and must adhere to all BSL-1 precautions, and additional
barrier protection devices such as face and eye protection, gloves, and lab
coats or gowns must be used. Most microbiology research, clinical, and
teaching laboratories maintain BSL-2 containment standards.
Biosafety level 3:
This level is applicable to clinical, diagnostic, teaching, research, or
production facilities in which work is done with indigenous or exotic
agents which may cause serious or potentially lethal disease. It includes
various bacteria, parasites and viruses that can cause severe to fatal
disease. For example : rabies virus , West Nile virus, Mycobacterium
tuberculosis and Chlamydia psittaci. The laboratory should be fitted
with special features such as negatively pressurized rooms and air filters
to prevent accidental release of the pathogen from the laboratory.
Biological safety cabinets are required. work must not be done on the
open bench.
Biosafety level 4
laboratories are designed for maximum containment of life-threatening
pathogens that have a high probability of transmission by aerosols and for
which there is no effective immunization, treatment, or cure. When
dealing with biological hazards at this level the use of a positive pressure
personnel suit, with a segregated air supply, is mandatory. The entrance
and exit of a Level 4 bio lab will contain multiple showers, a vacuum
room, an ultraviolet light room, autonomous detection system, and other
safety precautions designed to destroy all traces of the biohazard.
Multiple airlocks are employed and are electronically secured to prevent
both doors opening at the same time. All air and water service going to
and coming from a Biosafety Level 4 (P4) lab will undergo similar
decontamination procedures to eliminate the possibility of an accidental
release.
Examples of pathogens: hemorrhagic fever viruses ,drug-resistant
Mycobacterium tuberculosis.