How shall we work with microorganisms ? AFS 2005:01 ”Provisions from the Swedisk Work Environment Authority ” (AV) h

1. Howshallweworkwithmicroorganisms? AFS 2005:01 ”Provisions from the SwediskWork Environment Authority” (AV) http://www.av.se/lagochratt/afs/ Ingrid Bölin Dep. Microbiology and Immunology, GU

2. The provisions from 2005 wererevised 2012 and nowincluderegulations for workwithblood and otherbody fluids. The revised version will come into force May 1 2013. AFS 2012:17 Mikrobiologiska arbetsmiljörisker – smitta, toxinpåverkan, överkänslighet Arbetsmiljöverkets föreskrifter om ändring i Arbetsmiljöverkets föreskrifter (AFS 2005:1) om mikrobiologiska arbetsmiljörisker – smitta, toxinpåverkan, överkänslighet

3. Content Risk classes and risk assessment 2. Safetyprecautions 3. Notification/permission 4. Geneticallymodified microorganisms (GMM)

4. Definitions in AV provisins: Biological agents /influencingfactors a) Microorganisms, i.e., microbiologicalunitscapableofproliferation or transfer ofgenetic material b) Cellculturesofmulticellular organisms c) Virus and prions, d) Human parasites e) Componentsor substancesproduced by agents in a) to d) in connectionwiththese agents.

5. Risk class 1 • Risk class 1 comprisesnormally non-pathogenicbiologicalagents and non-pathogenicstrainsofpathogenic species or viruses. • Included in risk class 1 arealsononpathogenicbiological agents thatcan cause disease in otherways, like allergicreactions or toxicreactions. Exampleofriskclass 1: non-pathogenicEscherichiacoli

6. Risk class 2 • Biological agents thatmay cause diseaseswhichcan be cured or prevented or normallyareself-limitedwithoutserioussequelae. • Virusesthathavebeen isolated from humans and do not belongto a higher risk class. • Includedarealsobiological agents suspectedto cause cancer in humans, butwith a low risk after exposure, e.g. manyotherconcurrentfactorsareneeded. Example:Streptococcuspyogenes, Campylobacter-species

7. Risk class 3 • Included in risk class 3 areinfectious agents with a risk ofseriousconsequencesafter exposure, i.e.,causingseriousillnesswithlimitedpossibilities for cure or prevention or beinghighlycontagious. • Biological agents whichmay cause cancer and where the probabilityof cancer development is highafter exposure. Exempel: Mycobacteriumtuberculosis

8. Risk class 4 • Risk class 4 containsbiological agents with a veryhigh risk ofseriousdiseaseafter exposure. It may be a combination of a serious or deadlydisease, with no cure or prevention, and the risk ofepidemic transmission of a highlycontagious agent. Exempel: Ebolavirus • If a biologicala agent may cause cancer in humans withoutanyconcurrentfactorsinvolved it is alsoclassified as 4.

9. Swedish Institute for CommunicableDisease Control (Smittskyddsinstitutet,SMI) harbours the only P4 laboratory for workwith risk class 4 microorganisms in the Nordic countries. =safetylevel 4. Workwith risk class 3 microorganismsalsorequires special containmentlaboratories. Permission from AV is needed for workwithboth the highest risk classes.

10. Notification and permission 28 § The firsttime an employerintendstouse an infectious agent AV should be notified no later than 30 daysbefore the workwill start. Major changesthatwillaffect the risks should be reportedwithin a monthaftertheyhavebeenimplemented. Arbetsmiljöverket, Distriktet i Göteborg (Västra Götalands och Hallands län) Rosenlundsgatan 8, Box 2555, 403 17 Göteborg. Tel 031-743 72 00 Telefax 031-13 50 60 e-post: goteborg@av.se 29 § The followingactivitiesareonlyallowedafterpermission from AV. 1. Workwith risk class 2 agents in culturevolumesexceeding 500. 2. Workwith, or storageof risk class 3 or 4 biological agents.

11. Risk assessment and documentation • The roleof the employer is toassess the risks for diseases and accidents in the workplace. • Risk assessmentsshould be donetoidentifymicrobiological risks and necessarypreventivemeasures. • Adequatecompetences for risk assessementshall be available. • Writtendocumentation is needed. • Risk assessmentshaveto be renewed at regular intervals.

12. Factorsthatmayinfluence the risk assessment • The propertiesof the infectious agent: infectivedose, airborne transmission, availabilityofantimicrobialdrugs or vaccines and possibletoxic or allergic risks. • Procedureswithhigh risk: creationof aerosols or risk of spill. • The design of the workingenvironment. • Staff knowledge and training .

13. Protectivemeasures Four different levelsofprotectivemeasuresaredescribed in the provisions. The risk assessment and risk classof the agent decideswhichlevelthatshould be applied. Goodmicrobiologicalpracticeshould be employed : Cleanliness and order. Hand wash and/or disinfectionafterfinishingwork. No jewells on hands or hair or shawlshangingloose. No eating, drinking or working in a waythatmaycontaminate the mouth. Avoid spill, splashes and creationof aerosols. Avoidusingsharpitems. Useclosed containers for contaminatedglassware, etc. Useprotectiveclothing (labcoat).

14. Protectionlevel 2 Appropriatewarningsignsshould be displayed at entrences or at workplaceswherebiological agents are. The design of the signsshould be ”biologicalhazard” with information of risk class or protectivelevel and additionalnecessary information. The biological agents should be stored in a safeway. Protectiveclothingshould be wornwhenworkingwith risk class 2 agents. Protectiveclothes and other personal protectiveequipment must be removedwhenleaving the restricted area.

15. Protectionlevel 2, cont. Premises, furnishing and equipment. The premisesshould be demarcated. Thismeansthat the roomsshould be physically separated from otheractivitieswithwalls and doors. Access to the premisesshould be restricted (locked doors, acesscards for qualifiedpersonnel). Washbasinswith hand desinfectant. The tapsshould be possibletousewithout hands. Benches and floorsshould be resistanttodisinfectants. The rooomsshould be rodent and insectproof.

16. Protectionlevel 2, cont. Autoclaveshould be available. Autoclave must be in the vicinityof the workplace, normallylocated in the same building. Proceduresthatmay cause aerosol formation should be done in a microbiologicalsafetyhoodequippedwith HEPA-filter. The functionof the hoodshould be checkedregularily.

17. Protectionlevel 2, cont. Decontamination Decontaminationshould be doneto an extentthatpreventsbiological agents to cause disease and as soon as possiblewithsuitablemethods . The methodsusedshould be designed as toavoidmicrobiological air pollution or otherhealthhazards. Examples ofmethods: Autoclaving (should be usedbeforewashing-upofglassware.) Alcohol Virkon (for viruses) Iodine or chlorinecompounds

18. Protectivelevel 2, cont. Waste disposal Contaminatedwasteshould be handled and transferred toothercompanies or organisations accordingtoroutinesmade in advancetoavoidhealth risks. Necessary information about the waste, the risks associatedwith handling and the appropriatepreventivemeasuresshould be given to the personnelreceiving the waste. Containers should be leakproof and durable toavoidspreadof the content.Thecontentshouldalso be clearlymarked on the containers. Microbiologicalwastemay in somecase be sent for burning in special boxes for risk wastewithoutpreviousautoclaving.

19. Information and instructions 14 § The employershouldensurethat the employeeresponsible for workwithbiological agents and all employeesthatmay be exposedtomicrobiological risks haveappropriateeducation and sufficientknowledgeof the infectious agents used in the workingplace. All personnelinvolve in workwiththese agents should be given information on the risks and howtoavoidthem. 15 § The employerhavetoensurethat the employeesare given writtensafetyinstructionsand that the information has beenunderstood and that the instructionsarefollowed.

20. If an accidenthappens….. • The employer is requiredtohaveroutines for: • Reporting and documentingunwanted events and ill-health. • Takingmeasuresto limit the consequencesofunwanted events. • Cooperating withpeopleinvolved in order toinvestigate the cause ofunwanted events or ill-health. • Implementingmeasurestoavoidthat the same thingwillhappenagain.

21. Containeduseofgeneticallymodifiedmicroorganisms (GMM)AFS 2011:2 Containeduse: when organisms aregeneticallymodified or grown, stored, transported, destroidor ifsuchmodifedorganisms are handled in otherways and special cointainmentsmeasuresareusedtopreventthem from contactwith the general public. GMM: a microorganismwhich has beengeneticallychanged in a waythatdoes not occur in nature.

22. GMM The provisions on GMM arebased on the provisions for microbiologicalworkingenvironment risks, buthave a broaderscope. In addition to risks regarding human healththese provisions alsoincludes: Risks for diseases in animals and plants. Risks as a consequenceofspreadof GMM to the environment. Risks as a resultof transfer ofgenetic material tootherorgansisms. Gene technology is alsoregulated by otherlaws (Miljöbalken).

23. GMM Workwith GMM is classifiedaccordingto the requiredcontainmentlevel F-activity Workwith GMM withneglectable or no risk for humans or the environment. L-Activity Low risk Notificationto AV. R-activity Medium or high risk Permission required from AV!

24. GMM Risk assessment What kind ofgenetic material has beeninserted? Whatare the propertiesof the receiving organism? Will the new constructchangetheseproperties? Whatare the risks if the GMM is releasedinto the environment? Special demands for useof GMM in plants.

25. Referenser och användbara sidor • WHO Biorisk management: Laboratorybiosecurityguidance(2006) • WHO Laboratorybiosafety manual(2004) • CWA Laboratorybiorisk management standard(2008) • EU-direktiv 2000/54/EC (2000) • www.gmo.nu Biotechnologylegislation in Sweden withlinksto different authorities.