Download Biosafety in research laboratories

Elisabeth Haggård, Feb 2012. Biosäkerhetskommi;én Biosafety in research laboratories
• Responsibilities
• Biological agents
• GMM/GMO
• Blood or blood products
• Animal products
• Biosafety measure
Responsibility at the country level:
1.  Occupational heath law (Arbetsmiljölagen)
2.  Environmental law (Miljöbalken)
Ministry/authorities make specific regulations (föreskrifter)
• Work Environment Authority (Arbetsmiljöverket)
• Board of Health (Socialstyrelsen)
• Authority for Protection of the Environment (Naturvårdsverket)
• Chemical Inspectorate (Kemikalieinspektionen)
• Board of Agriculture (Jordbruksverket
Responsibility at the University level
The headmaster (rektor)
• Has an Environmental and Health Protection Committee (arbetsmiljönämnd)
• Has a Biosafety committee since may 2009
Elisabeth Haggård, professor Genetics
Katharina Pawlowski, professor Botany
Åke Wieslander, professor Biochemistry
Raad Askar, Veterinarian
Mats Hansson, safety engineer
Mattias Wadsten, head of safety at SU
• Delegates the responsibility to the prefects
Responsibility at the Department level:
• The prefect, delegates to the group leaders
Responsibilities of the group leader:
• Know the hazards and risks in the lab
• Make risk assessment
• Educate all personnel
• Provide protective equipment
• Control that the work performance is safe
• Check available emergency routines
Your responsibility:
• Be fully aware of all risks
• Be fully aware what protective measures are required
• Comply fully with applicable rules
• Report incidents and accidents
• Educate coworkers
• Be aware of emergency routines
Biological agents
The most important regulations for the research labs
are:
AFS 2005:01- Microbiological risks, toxins and
oversensitivity
AFS 2011:02/2000:5 – Use of genetically modified micro
organisms
AFS 1986:23 – Handling of blood and blood products
(only in Swedish)
Available as PDF from http://www.av.se/lagochratt/afs/
Laboratory infections.
Sulkin and Pike (1951-75) Active surveillance of 5.000 laboratories:
3.921 cases, 4.1% mortality (161 persons!).
Laboratory category
Research Diagnos=c Biological product Teaching Unspecified Number
2307
677 134 106 697 percentage
58.8 17.3 3.4 2.7 17.8 Known accidents or incident for only 18% of them • 25% needle sticks
• 27% spills
Routs of infection in the lab:
• Object to person Contact with media/materials
• Inhalation –air borne
droplets (aerosol), dust)
• Inoculation accidental injections, glass-breaks
splash to oral/mucus membrane
Routs of infection from the lab:
• Person to person • Ventilation –air borne
droplets (aerosol), dust
• Waste
• Waste water Factors to consider in
classification of agents:
• Mode of transmission
• Pathogenicity of the agents
• Availability of preventive/therapeutic
measures (antibiotics, vaccination)
Risk Group I
• Low individual and community risk
• A microorganism unlikely to cause human or animal disease
Ex. E. coli K12 and most cell cultures
However, some human cell lines or tumors are contaminated
with human pathogens*
Agent
No of samples tested No of positive samples
Prevalence %
HIV
1078
HepB
1078
HHV6 (Herpes virus) 268
HHV8
268
EVB (Epstein Barr virus) 268
9
8
3
1
4
0.83
0.74
1.12
0.37
1.49
Total
25
2.31
1078
* According to RADIL www.radil.missouri.edu
• Risk Group II
• Moderate individual risk, limited community risk
• A pathogen causing human or animal disease
unlikely to be a serious hazard to laboratory staff or
the community
• May cause serious infections but effective treatment is available and risk of spread is limited
Ex. Staphylococcus, influenza virus, scrapie (prion)
Risk Group III –
Biosafety lab level 3 required
• High individual risk, low community risk
• A pathogen causing serious human disease but not readily
transmitted to others
• Effective treatment is available Ex. Mycobacterium tuberculosis, Shigella dysenteria, EHEC
Risk Group IV –
Biosafety lab level 4 required, one at SMI
• High individual and community risk
• A pathogen causing serious human or animal disease, readily
transmitted between individuals
• Effective treatment
usually not available Ex. Ebola virus Demands on laboratories at biosafety level 2-4
Isolated location Entrance through airlock
Observation window Separated ventilation system
with HEPA filter outlet Neg pressure Personal shower Safety cabinets
Closed for contamination
decontamination
-of solid waste, autoclave
-of waste water
Reserve electricity Alarm for safety installations
2
Delimited
No Optional 3
Yes Yes Yes 4
Separate building
Yes
Yes
No No Optional Optional Optional Yes Yes Optional Yes Yes Yes + HEPA air inlet
Yes
Yes
Yes
Yes
Yes Optional No No Yes in lab
Yes
Optional Yes Yes in lab
Yes
Yes
Yes
Report/permission for using pathogenic organisms
First time use of class 2 organisms must be reported
to the local Work Environment Authority (Arbetsmiljöverket)
30 days before the work is initiated.
Class 3 organisms requires permission from the Work
Environment Authority (Arbetsmiljöverket) and special lab
and are time limited.
GMO Genetically Modified Organism
In the Environmental law (Miljöbalken) chapter 13 is an organism defined as: A biological unit that can multiply or transfer genetic material (3 §) The definition of an organism includes besides plant, animals etc., also for example seeds and pollen.
A genetically modified organism is an organism whose genetic material has been changed in a way that does not occur naturally by mating or natural recombination (4 §) Regulations for using GMO
(SFS 1994:901)
Depends on activity Contained use of:
• Microorganisms • Water living organisms • Other organisms Field experiments:
• Microorganisms, nematodes,
spiders, insects Water living organisms
• Trees • Other organisms On the market
Authority
Work Environment Authority Board of Fisheries (Fiskeriverket)
Board of Agriculture (Jordbruksverket)
Chemical Inspectorate Board of Fisheries
Forest Agency (Skogsstyrelsen)
Board of Agriculture Several authorities
GMM Genetically modified microorganism. In SFS 2000:271 is a microorganism defined as all microbial
units, cellular or not cellular that can multiply or transfer genetic
material. GMM is defined as a microorganism whose genetic material has been changed in a way that does not occur naturally by
mating or natural recombination
Includes: • Bacteria, virus, viroids, blue-green algae, molds, protozoa,etc.
• RNAi if inserted via a vector, but not if inserted by a liposome
• Cell lines from plants or animals
• Cells microinjected with DNA
• Cell fusions or hybridization techniques generating living cells
with new combination of genetic material that might be
dangerous for health or environment.
Examples of what is not considered as GMM
• Microorganisms generated by mutagenesis
• Microorganisms generated by cell fusions or prokaryotes
exchanging genetic materials by known natural processes
• Cell fusions or hybridization techniques generating living cells with new combination of genetic material that is not
dangerous for health or environment.
GMM
All work with GMM must be reported to or have
permission from the Work Environment Authority
(Arbetsmiljöverket)
They have forms that should be filled in.
Different forms depending on the safety level of the activity
Level of activity
Risk
Safety level
F
Negligible
1
L
Low
2
R
High
3
Riskassessment determines the required biosafety level Unmodified microorganism
Risk class
Safety level
1
1 E. coli K12
Cell cultures
2
2 3
3 GMM
Risk class
Safety level
Category
1
1 or 2 (3)
F or L (R)
2
2 or 3 (1)
L or R (F)
3
3 (2)
R(L)
F and L activities have to be reported
R activities requires permission
Handling blood or blood products.
All contacts with blood that has not been purified from infectious agents for example by heat treatment is a potential risk.
Possible infections
Most common: Hepatitis B, vaccine available, risk of infection by inoculation 10-30%
Other diseases with a risk are:
HIV, no vaccine, risk of infection by inoculation 0.3%
Hepatitis C, no vaccine, risk of infection by inoculation 3%
Less common are:
Hepatitis D&G, HTLV 1 and 2, Creutzfeldt-Jakobs disease
Animal products or ”side-products”.
Definition by Board of Agriculture (Jordbruksverket):
Animals, tissues, products not intended for food, i.e. even proteins >10 kD, Ab
not purified by affinity chromatography and sera. Should be considered as potentially infectious, and thus incinerated.
However, CE marked products can be regarded as non-infectious and wild
caught Swedish animals and fish believed to be uninfected.
All imports to SU must be reported every half year to the Board of Agriculture.
Three categories, with examples from each:
1.  Animals or materials from animals known or suspected to be infected with
pathogens.
2.  Animals or spills from animals not of category 1. Research animals that has
not been used for research.
3.  Products of animals originally intended for food.
Protective equipment:
Personal protective equipment:
• Gloves
• Gown
• Mask/goggles/face shield
Safety equipment:
• Cabinets, class I-III
• Automatic pipetting device
• Safety (locked) centrifuges
Class I biosafety cabinet
Class II biosafety cabinet (type A2)
B-types connection to building exhaust system required
Clas III biosafety cabinet
Connected to building exhaust system required
Good Microbiological Technique GMT
Specimens are handled safely.
No mouth pipetting is permitted. Pipettes and pipetting aids are used safely. Dispersal of infectious materials is avoided. Contact of infectious materials with skin and eyes is avoided. Ingestion of infectious materials is avoided. Separation of serum is carried out safely. Centrifuges are used safely. Homogenizers, shakers and sonicators are used safely. Tissue grinders are used safely. Refrigerators are maintained and used safely. Ampoules containing infectious materials are opened safely. Infectious materials are stored safely. Precautions are taken with blood and other bodily fluids. Specimens and infectious materials are shipped safely. Appropriate disinfection and sterilization are carried out. Gloves are worn for procedures that involves contact with blood or infectious material. Hands are washed between procedures and prior to leaving laboratory. Laboratory gowns are worn for work in laboratory. Closed-toed shoes are worn for work in laboratory. Storage of food or drink in the laboratory is prohibited. Eating, drinking, or smoking in the laboratory is prohibited.
Last but not least!
All incidences/accidents should be reported!
Stockholm University has an easy way of
notifying incidences and accidents. Go to
SAMIR, where you will find information and a
form to fill in.