Health & Safety

- You should think of “safety” while doing any practical work to avoid
harming yourself or others.

- Safety is considered everyone responsibility.

COSHH

Control of Substances Hazardous to Health

- Are group of “regulations” including specific legal requirements for risk

assessment when using hazardous chemical or biological agents.

- These regulations can be applied through ‘approved’ codes of practice for

the control of hazardous substances, carcinogens and biological agents,
including pathogenic microbes.
 Hazard & Risk

Hazard: the ability of a substance or biological agent to cause Harm.

Risk: the pRobability of a substance or biological agent to cause harm in a specific

situation.

How to differentiate between Hazard and Risk:

One of the Hazards associated with water is drowning.

However, the Risk of drowning in a few drops of water is minimal !!
 Warning
Signs

Radiation hazard

Toxic
Laser Hazard Flammable- oxidizing

US protectors
Low temperature
Emergency eye wash Explosive
 Exposure Routes to Harmful substances

1- Skin Absorption
Some materials are
2- Inhalation
capable of penetrating
intact, healthy skin as Of gases, vapours, aerosols,
phenol fumes

4- Injection
3- Ingestion
Accidental on contaminated food, drink
Biological Hazard

- The most obvious risks when handling microbial cultures are those due to
ingestion or entry via a cut in the skin.

- All cuts should be covered with a plaster or disposable plastic gloves.

- A less obvious source of hazard is the formation of aerosols of liquid

droplets from microbial suspensions, with the risk of inhalation, or surface
contamination of other objects.
 Classification of microbes on the basis of hazard,
UK Advisory Committee on Dangerous Pathogens
(ACDP)

Hazard group Comments

1 Unlikely to cause human disease.

2 May cause disease

- Possible hazard to laboratory workers / minimal hazard to
community.

3 May cause sever disease

- May be a serious hazard to laboratory workers / may spread to
community.

4 Causes sever disease

- Is a serious hazard to laboratory workers / high risk to
community.
Laminar flow cabinet
- Prevent airborne contamination e.g. when
preparing media or sub-culturing microbes or
tissue cultures.

- Sterile air is produced by passage through a

high efficiency particulate air (HEPA) filter,
which is then directed towards the working area,
either horizontal towards the operator or
downwards.

- No protection to the worker from contamination

and so, must not be used with pathogenic
microbes.
 Bio-safety Labs
4 bio-safety levels (BSLs) according to different laboratory techniques, safety
equipment, and design, depending on the types of agents being studied:

BSL-1 labs: used to study agents not known to consistently cause disease in healthy
adults.
- Basic safety procedures - no special equipment or design features.

BSL-2 labs: used to study moderate-risk agents that pose a danger if accidentally
inhaled, swallowed or exposed to the skin.
- Safety measures: gloves - eyewear - hand washing sinks - waste decontamination
facilities.

BSL-3 labs: used to study agents that can be transmitted through the air and cause
potentially lethal infection.
- Safety measures: Gas-tight enclosure - specialized ventilation system - clothing
decontamination - sealed windows.

BSL-4 labs: used to study agents that pose a high risk of life-threatening disease for
which no vaccine or therapy is available.
- Safety measures: wearing full-body, air-supplied suits - shower when exiting the
facility - occupy safe, isolated zone within the building.
 Substance Hazard Safety

SDS ‘sodium dodecyl -Irritant Wear gloves

sulphate’ -Toxic

NaOH ‘sodium - sever irritant Wear gloves

hydroxide’ - highly corrosive

- irritant/corrosive Wear gloves

Isopropanol - highly flammable No naked flames
- potential carcinogen

- highly toxic Wear gloves

Phenol - skin burns Use in fume hood
- potential carcinogen

- volatile and toxic Wear gloves

Chloroform - irritant/corrosive Use in fume hood
-potential carcinogen
 Ethidium bromide and COSHH

- Ethidium bromide (EtBr) is widely used for detection and quantification of

DNA/RNA, but is an extremely hazardous material.

- The ready prepared ethidium bromide solutions decrease the risk associated
with handling the solid when preparing the stock solutions.

Risks associated with ethidium bromide:

R 22 Harmful if swallowed
R 23 Very Toxic by inhalation
R 68 Possible risks of irreversible mutagenesis effect: intercalating agent and
frame-shift mutagen.
*R: indication of risk associated.

- Inexpensive, but the hidden costs of disposal are very high. charcoal
filters, operator time to filter the waste running buffer, and bagging of solid
waste for incineration.
Disposal of ethidium bromide:
- Aqueous solutions of ethidium bromide (>1.0µ g/ml) may be disposed of in
one of two ways:

1- Adsorbed onto an ‘ion exchange column’ specifically for this purpose, These
columns will adsorb at least 300mg of ethidium bromide; the waste liquid may
then be discarded to drain. Then, the columns should be double-bagged in
plastic bags and disposed of as for solid waste.

2- Adsorbed onto activated charcoal at a rate of 100mg charcoal to 50mg ethidium

bromide. The mixture should be left overnight before filtering off the solid,
which should then be double-bagged in plastic bags, then disposed of as for
solid waste. And the waste liquid can be discarded to drain.

Solid ethidium bromide waste (e.g. gels, contaminated paper towels, etc)
should be placed into a suitable, leak-tight container and then into a yellow bag
and treated as clinical waste for incineration.
SYBR Safe Green

- An alternative of the ‘ethidium bromide’.

- No acute oral toxicity and greatly reduced genotoxicity.
- Non hazardous waste.
- No special handling, storage or disposal.
-Can be excised from gels under non-UV light, which reduces UV-induced
cross-linking of the DNA you want to further manipulate.

But more expensive!!

References:

Books:
- Reviewed in Practical skills in biology, 2003