Safety Instruction

Applies to practical course „Microbiology“

Bachelor Curriculum „Water Science“

Biofilm Centre, University of Duisburg-Essen,

Campus Essen

Building S05 T02, Laboratories A32, A35 and A55

10 March 2017

Dr. Jost Wingender

According to

- Ordinance on Biological Agents (§ 14 Biostoffverordnung)

- Ordinance on Hazardous Substances (§ 14 Gefahrstoffverordnung)

Regulates safety and health protection when

working with biological agents.

Provides an institution with a framework for a

risk assessment.

Commits the employer to provide operation

instructions.

Demands a briefing on safety instructions for

employees at least once a year and prior to

commencement of work.

Ordinance on Biological Agents

Biohazard

What are biological agents?

Definitions according to Ordinance on Biological Agents

Biological agents are microorganisms, cell cultures and

endoparasites, including their genetically modified forms, as well as

agents associated with transmissible spongiform encephalopathy,

which can endanger human health through infections, transmitted

diseases, formation of toxins, sensitizing, or other health damaging

effects.

Microorganisms are all cellular and non-cellular microscopic and

sub-microscopic biological units, which are capable of transferring

genetical material, in particular bacteria, viruses, protozoa or fungi.

Biological agents are categorized according to the risk of infection into

four risk groups.

Relevant biological agents used in practical course belong to risk

groups 1 and 2.

Risk group 1: Biological agents which are unlikely to cause illness

in humans.

Risk group 2: Biological agents which can cause illness in

humans and present a danger to the employees; the spread of

the agents in the human population is unlikely; effective preventive

actions or treatment are usually possible.

(Examples: Escherichia coli, Pseudomonas aeruginosa)

Classification of biological agents into

risk groups

Authorization to work with pathogens

(Biological agents of risk group 2)

Dr. J. Wingender: authorization (according to the German

Infection Protection Act) to work with pathogens belonging

to risk group 1 and 2.

All employees are allowed to work with pathogens

belonging to risk group 1 and 2 under his supervision and

do not require any other permit.

Working with biological agents

Handling of biological agents according to the

Ordinance on Biological Agents:

- Isolation

- Cultivation

- Cell disruption

- Mixing and separation

- Transport

- Storage

- Inactivation and disposal

Potential transmission routes in the

laboratory

Potential routes of contamination or infection under

laboratory conditions:

- Inhalation Formation of aerosols by spilling of

liquids, mixing, vortexing.

- Resorption through skin Spilling of liquids, contact of hands with

or mucous membranes contaminated equipment, contact with

eyes, nose, mouth

- Ingestion Pipetting by mouth, splashes to mouth

- Inoculation Cuts, needle pricks

Safety rules for handling of biological

agents

- Keep work place clean and tidy. Remove chemicals and devices that are not

permanently used.

- Do not work alone, ensure visual contact to colleagues.

- Keep windows and doors closed.

- Wear protective clothes (lab coat, gloves, safety glasses) and sturdy shoes.

- Eating, drinking, smoking, chewing gum, etc. as well as storage of food or

tobacco is strictly prohibited.

- Do not pipet by mouth, use mechanical transfer devices.

- Keep work place clean and tidy. Remove chemicals and devices that are not

permanently used.

- Avoid formation of aerosols.

- Exercise caution when working with the naked flame (Bunsen burner).

- After work disinfect work place (Bacillol AF; protective gloves), will be done by

„lab service“ (participants of practical course).

- Before leaving the laboratory, disinfect hands (Sterillium) see hygiene plan.

- After contamination of skin with biological agents, thoroughly disinfect

contaminated skin area (skin disinfectant, Cutasept F).

- After contact of biological agents with the eyes, rinse eyes with plenty of water

(at least 10 – 15 min); use eye shower.

- Any kind of accidents need to be reported to the responsible persons (Prof.

Meckenstock, Dr. Wingender).

- In case of pregnancy, report immediately to Prof. Meckenstock.

- Consider hygiene plan!

Safety rules for the handling of

biological agents

Hygiene plan

Hygienic hand disinfection

Wash basin with dispensers

of hand disinfectant and

liquid soap

Hygiene plan

Hygiene plan

What are hazardous substances?

Hazardous materials are substances or preparations have

the following properties:

• explosive,

• oxidizing,

• extremely flammable,

• highly flammable,

• flammable,

• very toxic,

• toxic,

• harmful,

• caustic, corrosive

• irritant,

• sensitizing,

• carcinogenic,

• teratogenic,

• mutagenic

• dangerous to the environment

• biohazardous

or

materials from which hazardous or explosive substances or

preparations can be produced or liberated

GHS - Globally Harmonized System

Hazardous substances

• Nine labels

• Hazard statements (H phrases)

• Precautionary statements (P phrases)

Regulation on Classification, Labelling and Packaging of Substances and

Mixtures

explosive flammable oxidative Pressurized

gases

corrosive acute toxic Acute toxic

Mutagenic

Carcinogenic

Reproductive toxicity

respiratory sensitizing

Aspiration hazard

irritant,

sensitizing

toxic

environment

ally

hazardous

Example: disinfectant Bacillol AF

Intake routes of hazardous substances

Possible intake routes into the human body in the laboratory:

- Inhalation Dust or powder during weighing,

aerosols caused by spilling of

liquids, mixing, vortexing.

- Resorption through skin or Spilling of liquids, contact of

mucous membrane hands with contaminated

equipment, hand-eye/nose/mouth

contamination

- Ingestion Pipeting with the mouth

Splashes to the mouth

- Inoculation Cuts, needle prick

Basic rules for handling

hazardous materials

Own responsibility to inform yourself prior to commencing work with hazardous

materials (e.g. Ordinance on hazardous substances, operating instructions, safety

data sheets, internet)

Wear protective clothes (lab coat, gloves, safety glasses) and sturdy shoes

Eating, drinking, smoking, etc. is prohibited

No storage of foods or tobacco in the laboratory.

Use hazardous materials only in quantities necessary for the experiments

Do not store hazardous material in containers, which are normally used to

store food! Danger of confusion!

Every storage container needs to be labelled with its name and danger labels

Labelling needs to be permanent with water proof pens; direct labelling with felt

markers or pasting over existing labels is not permitted!

Do not pipet by mouth, use mechanical transfer devices

Waste prevention and disposal

1. Use hazardous materials only in quantities that are necessary for

the experiment

2. Waste needs to be collected or packed and labelled in the

appropriate storage containers.

3. IDEXX-containers: Only for Colilert-18/Quanti-Trays waste.

4. Collect broken glass in the respective glass containers (non-

contaminated/contaminated).

Precaution and safety

devices

Inform about:

• Escape routes (do not block escape

routes)

• Emergency call 112

• Location of emergency facilities (regularly

control the functional capability, report)

- first aid boxes

- emergency shower

- eye wash

-emergency shutoff devices

• Fire protection equipment

- fire alarm

- fire extinguishers

- official fire regulations

Behavior in dangerous

situations

• Stay calm

• Protecting persons prior to

equipment protection

• Protecting people prior to fire

protection

• First inform others (fire alarm),

then fight fire

• protect your own safety

• Provide first aid

Behavior in dangerous situations

• Accidents/irregularities - immediately inform others (Prof.

Meckenstock etc.), project leader, warn colleagues.

• All injuries/accidents have to be recorded in the accident book

(“Verbandsbuch“); records have to be stored at least 10 years)

• Wipe up spilled suspensions containing biological material,

using tissue paper (autoclave)

• Desinfect contaminated equipment and surfaces with Bacillol AF

(15 min incubation)

• After skin contact with biological agents desinfect with skin

disinfectant.

• Bring injured persons out of the danger zone and provide fresh

air.

• Extinguish burning clothes.

• If necessary, use emergency shower. If contaminated, clothes

have to be removed before; wash with soap and water.

Behavior in dangerous situations

• Stop bleedings and apply a bandage (use examination

gloves)

• If eyes are contaminated with biological material, wash out

the eyes with huge amount of water (at least 10 min); use the

eye wash station

• After intensive contact (e.g. ingestion), consult doctor

• Do not leave injured person alone.

• Collect information for the doctor.

(vomit, chemicals and other details, e.g. safety data sheets)

• Autoclave contaminated clothes.

Emergency telephone numbers

3333 In-house emergency call connection with switchboard

(inform someone from the switchboard at the latest after all rescue measures

are done)

112 Fire Department, emergency doctor, rescue service

- from all telephones inside the university at each time possible

- free emergency call from public telephones

- Mobile phone

Meeting place: lawn in front of building S05