Safety, Health and Environment (SHE) Khairul Farihan Kasim School of Bioprocess Engineering UniMAP ~kfk~

Biosafety In Biopharmaceutical Facilities ~kfk~

Biosafety Biosafety is a term used to describe efforts to reduce and eliminate the potential risk resulting from biotechnology and its product. The potential risks can be grouped into the three areas of sustanable development: Environment concerns (release to environment, biodiversity) Social concerns (threat to human health from new allergens in the food system) Economic concerns. ~kfk~

Where Hazard Come From? Physical: Due to wrong operation of Equipment Due to miss-use of Equipment Due to lack of safety precaution during opearation Chemical: Due to use of flammable or Biohazard chemicals outside Fume Cupboard Not using suitable cloths, Gloves, Eye glass, etc…. Biological: Emergency cases (leakage of hazard chemicals from broken tools & equipments) Due to wrong use of biomaterials -Use of MO in less safe lab. for its group - Wrong decontamination protocol ~kfk~

Before Running Experiment! Types of biological materials used in experiment The mode of modification applied and methodology Type of Chemicals used and precautions of these chemicals The different types of expected and unexpected biomaterials arised Documents on the waste disposal regulation of the chemicals and biomaterials in each step -Host (BI, BII, BIII, etc…., GRAS?) - Vector NO RADIOACTIVE MATERIALS ARE ALLOWED IN Biomanufacturing facilities ~kfk~

Inside the Biotechnology Factory (Interferon Production Facility) Working with hazardous chemicals (non-radioactive) Working with B1 and B2 microorganism Working with GMO Working with equipments with special requirements. (UV lamps, sonicators, etc….) ~kfk~

Recombinant MO If the wild strain considered as GRAS, its recombinant is not GRAS anymore. The recombinant strain should not have any known combinant of pathogenicity, high colonization ability, and high genetic transfer competency. The recombinant strain should not able to survive and colonize in the environment, and should not able to transfer any part of it’s genome to indigenous population. Recombinant E. coli K-12 strains and their plasmidless hosts are unable to establish in environments consistent with various deliberate release scenarios. Moreover, those strains are non conjugating and apparently incapable of transferring genes to other organisms. ~kfk~

Biosafety Levels BS Agents Practice Safety Equipments (Primary Barriers) Facilities (Secondary Barriers) L1 & L1-LS Not known to consistent cause disease for healthy personnel Standard Microbiological Practice Standard BSC Class I Open bench top sink required L2 & L2-LS Associated with human disease, hazard , percutaneous injury. Mucous. Recombinant MO BSL- 1 practice plus. Limited access Biohazard warning signs Sharps precautions Waste decontamination policy Standard BSC class II BSL-1 plus Autoclave available. L3 & L3-LS Indigenous or exotic agents with potential for aerosol transmission, disease may have serious or lethal consequences. BSL-2 practice plus: Controlled access Decontamination of all wastes Decontamination of lab. Clothing before laundering BSL-2 plus - Physical separation from access corridors. Self closing, double door accesss Exhausted air not recirculated Negative airflow in the laboratory L4 Dangerous/exotic agents which pose high risk of life-threatening disease BSL-3 practice plus: -Clothing change before entering Sower on exit All material decontaminated on exit from facility Standard BSC class III or BSC class II in combination with full-body air supplied positive pressure BSL-3 plus - Separates building or isolated zone -dedicated supply and exhaust vacuum ~kfk~

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Extra Requirement for Good Large Scale Practice (GLSP) Implementation of a health and safety program. Suitably trained personnel and written operational procedures. Facilities, Equipment, protective clothing and practices appropriate to risk. Regulatory compliance with regard to discharged to the environment Minimization of aerosol generation to prevent adverse risk to employee health. A spill control plan within the emergency response plan. GLSP should preferably be designed to one level higher than the minimum acceptance biosafety level. ~kfk~

Points of Potential Release of Controlled Material Shaft seals (vessels, pumps) Flanges (Pipes, Valves, Vessels) Points of Probe and sensor entry Perforated pipes, vessels, Equipment Rupture Disk Exhaust gases Tubing and hoses Pumps Sample points Gasket Leakage into cooling water Safety valves Waster treatment system Sanitary connections ~kfk~

Biohazard from Biomaterials Laminar Air – Flow (Biological Safety Cabinet) And Decontamination System ~kfk~

Biosafety Cabinet Type Face velocity (1fpm) Airflow Pattern Radionuclides / Toxic Chemicals Biosafety Level (s) Product Protection Class I open front 75 In a front:rear and top through HEPA filter No 2,3 Class II Type A 70% recirculated through HEPA; exhaust through HEPA Yes Type B1 100 30% recirculated through HEPA: exhaust via HEPA and hard ducted Yes (low levels/volatility) Type B2 No recirculation:total exhaust via HEPA and hard ducted Type B3 Same as IIA, but plena under negative pressure to room and exhaust air is ducted Class II NA Supply air inlets and exhaust through 2 HEPA filter 3,4 ~kfk~

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Decontamination Sterilization Disinfection Antiseptics The use of a physical or chemical procedure to destroy all microbial life, including large numbers of highly resistant bacterial spores The use of a physical or chemical procedure to virtually eliminate all recognized pathogenic microorganisms but not all microbial forms (bacterial endospores) on inimate objects A germicide that is used on skin or living tissue for the purpose of inhibiting or destroying microorganisms. ~kfk~

Decontamination Methods Heat Chemicals Radiation ~kfk~

Agent Selection Degree of microbial killing required Nature of item/surface to be treated Ease of use Safety Cost ~kfk~

Agent Efficiency Type of organism Number of organisms Amount of organic material Type & configuration of material to be treated Time & concentration or exposure pH Humidity Agent Efficacy ~kfk~

Chemical Agent Selection HLD-High level disinfection ILD-Intermediate level disinfection LLD- Low level disinfection Sporocides Kills all microorganisms except high numbers of bacterial spores Required 5-10 min. exposure Examples: aldehydes, hydrogen peroxide, paracetic acid -Tuberculocides Kills M. tuberculosis var. bovis and all vegetative bacteria, fungi, and most viruses -Require minimum 20 min. Exposure -Examples: phenolics, iodophores, chlorine compounds, alcohols. Hospitals germicides used for housekeeping Kills most vegetative bacteria and some fungi, but not M. tuberculosis var. bovis Require minimum 20 min. Exposure Examples: quartenary ammonium compounds ~kfk~

Decontamination Summary Sterilization Bacterial Spores B. Subtilis Mycobacterium MTB var. bovis Non-lipid Viruses Polio- Rhino- Fungi Cryptococcus sp. Candida sp. Vegetative Bacteria Pseudomonas sp. Staphylococcus sp. Salmonella sp. Lipid Viruses Herpes CMV HBV HIV 2.10 Decontamination Summary Sterilization HLD LLD ILD Sterilization HLD ILD LLD ~kfk~

Different Types of Wastes in Biopharmaceutical Factory Biohazard waste (Containing Process related biomaterials) Solid waste Chemical wastes Liquid waste Non-Process related wastes (wastes of non-production area) Not contaminated with Chemicals or Biomaterials of process Gas waste Waste Management System in Factory is Important ~kfk~

Waste Management System (Hazardous wastes) Solid and Liquid wastes Segreagation and separation of different types in suitable container Collection I Collection II Decontamination Release of safe waste* (house hold level) *, this step could be done by waste contractor (if available) ~kfk~

The role of Biosafety Committee in Biopharmacetical factory To Put A regulatory system to register and document all biomaterials Establish a mechanism to handle requests for permits for certain activities, such as release of GMOs into the environment through the national biosafety committee Establishing a mechanism for monitoring and inspections Putting system to provide information for personnel (Training, workshop, Biosafety Guidelines, etc…) Participate as consulting during the purchase of new equipments Reviewing the SOPs of all department (safety of chemicals, method and suitability of biosafety level to the area classification. Preparation of Biosafety Manuals ~kfk~

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