Biosafety in research laboratories
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1 Elisabeth Haggård, Feb Biosäkerhetskommi;én Biosafety in research laboratories Responsibilities Biological agents GMM/GMO Blood or blood products Animal products Biosafety measure
2 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
3 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
4 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
5 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
6 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
7 Laboratory infections. Sulkin and Pike ( ) Active surveillance of laboratories: cases, 4.1% mortality (161 persons!). Laboratory category Number percentage Research Diagnos=c Biological product Teaching Unspecified Known accidents or incident for only 18% of them 25% needle sticks 27% spills
8 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
9 Routs of infection from the lab: Person to person Ventilation air borne droplets (aerosol), dust Waste Waste water
10 Factors to consider in classification of agents: Mode of transmission Pathogenicity of the agents Availability of preventive/therapeutic measures (antibiotics, vaccination)
11 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
12 However, some human cell lines or tumors are contaminated with human pathogens* Agent No of samples tested No of positive samples Prevalence % HIV HepB HHV6 (Herpes virus) HHV EVB (Epstein Barr virus) Total * According to RADIL
13 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)
14 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
15 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
16 Demands on laboratories at biosafety level Isolated location Delimited Yes Separate building Entrance through airlock No Yes Yes Observation window Optional Yes Yes Separated ventilation system with HEPA filter outlet No Yes Yes + HEPA air inlet Neg pressure No Yes Yes Personal shower Optional Optional Yes Safety cabinets Optional Yes Yes Closed for contamination Optional Yes Yes decontamination -of solid waste, autoclave Yes Yes in lab Yes in lab -of waste water Optional Yes Yes Reserve electricity No Optional Yes Alarm for safety installations No Yes Yes
17 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.
18 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 )
19 Regulations for using GMO (SFS 1994:901) Depends on activity Authority Contained use of: Microorganisms Work Environment Authority Water living organisms Board of Fisheries (Fiskeriverket) Other organisms Board of Agriculture (Jordbruksverket) Field experiments: Microorganisms, nematodes, spiders, insects Chemical Inspectorate Water living organisms Board of Fisheries Trees Forest Agency (Skogsstyrelsen) Other organisms Board of Agriculture On the market Several authorities
20 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.
21 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.
22 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 F L R Risk Negligible Low High Safety level 1 2 3
23 Riskassessment determines the required biosafety level Unmodified GMM microorganism Risk class Safety level Risk class Safety level Category or 2 (3) F or L (R) E. coli K12 Cell cultures or 3 (1) L or R (F) (2) R(L)
24 F and L activities have to be reported R activities requires permission
25 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
26 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.
27 Protective equipment: Personal protective equipment: Gloves Gown Mask/goggles/face shield Safety equipment: Cabinets, class I-III Automatic pipetting device Safety (locked) centrifuges
28 Class I biosafety cabinet
29 Class II biosafety cabinet (type A2) B-types connection to building exhaust system required
30 Clas III biosafety cabinet Connected to building exhaust system required
31 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.
32 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.
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