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Transcript
Biological Hazards Examination Center for Laboratory Safety and Health Study Aims To understand the definition and characteristics of biological hazards (biohazards) To know the transmission modes of biological agents To learn the classification of biohazards To know the prevention and control methods of biohazards 2 Outline Unit 1 Biology Overview 1.1 Biological Classification 1.2 Definition of Biohazards Unit 2 Sources and Modes of Transmission 2.1 Sources of Biohazards 2.2 Modes of Transmission Unit 3 Adverse Effects 3.1 Infection 3.2 Allergy 3.3 Toxic effects 3.4 Others Unit 4 Biohazard Prevention and Control 4.1 Prevention – Classification of Infectious Microorganisms by Risk Groups – General Prevention Methods (Environmental Management & Personnel Management) 4.2 Control – Disinfection and Sterilization – Biological Safety Cabinets – Temporary Storage and Management of Biomedical Waste 3 Unit 1 Biology Overview 1.1 Biological Classification Non-cellular Organisms – Virus Prokaryotes – Archaea – Eubacteria Microorganisms Eukaryotes (Eukarya ) – Fungi Source: Wikipedia – Protista • Photosynthetic (plant-like) protists : algae • absorptive (fungus-like) protists: no specific name • ingestive (animal like) protists : protozoa – Plantae – Animalia 5 Virus 20-300 nm (1 nm=10-9 m) non-cellular nucleic acid (RNA or DNA ) enclosed in a protein coat (capsid) obligate parasites specificity e.g., Influenza viruses (Types A, B, C), SARS coronavirus, Rabies virus , Hantavirus, HIV, Foot-and-mouth disease virus Non-cellular Organisms 6 Influenza Virus and Novel Influenza A (H1N1) Virus Influenza viruses: type A (causing human influenza pandemics), type B, type C – Viral envelope containing two main types of glycoproteins, hemagglutinin (HA) and neuraminidase (NA) – The central core containing single-stranded RNA genome – Common influenza A subtypes: H3N2, H1N1, H5N1 (avian flu) Novel influenza A (H1N1) virus – A subtype of influenza A virus – H1N1 viruses not normally infect humans → reassortment of swine, avian and human flu genes → human contact with infected swine → person–to–person transmission →pandemics 7 Prokaryotes Archaea existing in harsh environments, such as hot springs, salt lakes and volcanic vents different structure of cell membrane and cell wall compared to bacteria e.g., halophiles, thermophiles, alkaliphiles and acidophiles http://lesliegottschalk.blogspot.com 8 Prokaryotes Bacteria cell structure 0.5-1μm × 2-5 μm (1 μm = 10-6 m) mostly free-living e.g., Staphylococcus aureus, Pneumococcus, Clostridium botulinum , Bacillus anthracis, Escherichia coli, Mycobacterium tuberculosis. 9 Prokaryotes Endotoxins a constituent of the outer membrane of Gram-negative bacteria lipopolysaccharide (LPS) released into the environment after lysis of the bacterial cell or during active cell growth Source: http://horseshoecrab.org/med/med.html 10 Prokaryotes Bacterial Exotoxins mostly from Gram positive bacteria proteins that a bacterium produces internally and secretes into its environment e.g., exotoxins produced by Clostridium tetani, Clostridium botulinum and Corynebacterium diphtheriae 11 Prokaryotes Bacterial Endospores Function: to resist harsh environment (e.g., heat, desiccation, UV radiation, chemical disinfectants) e.g., Bacillus anthracis Clostridium tetani https://wikispaces.psu.edu/display/ 110Master/Prokaryotes+II++Structure+and+Function 12 Eukaryotes Fungi similar to plants without chlorophyll releasing enzymes to digest organic compounds including molds, yeasts (2-10 μm) and mushrooms (larger size) unicellular or multicellular 13 Eukaryotes Mycotoxins fungal metabolites to aid competitiveness and success of colonization of a fungus over other microorganisms (fungi or bacteria) cytotoxic: destroying cell structures (e.g., cell membranes), interfering important process of life (e.g., RNA and DNA synthesis) examples: Aflatoxin , Ochratoxin A 14 Eukaryotes Fungal Spores sexual and asexual spores function – dissemination and reproduction characteristic – resistant Fig 1: Conidiogenous cells and chain of conidia of Aspergillus sp. http://digiku.nmns.edu.tw/4images/details.php?image_id=44 Fig 2: Halophytophthora spinosa (zoosporangium) http://digiku.nmns.edu.tw/4images/postcards.php?image_id=45 15 Eukaryotes Protista (Protoctista) Photosynthetic (plant–like) protists –Algae – Photoautotrophs, with chloroplasts Absorptive (fungus–like) protists – Chemoheterotroph: phagocytize organism or excrete enzyme in order to decompose and absorb organic compounds Ingestive (animal–like) protists – Protozoa – meaning “first animals” – Predatory or parasitic – e.g., Amoebas, Giardia , Cryptosporidium, Plasmodium, Paramecium, Foraminifera. 16 Eukaryotes Higher Plants Ingest, inhale or contact with plants or their products Inhalation: hay fever – anemophilous flowers – pollen → allergen – IgE binds to allergens → release of histamine and inflammatory mediators – Allergic rhinitis and allergic conjunctivitis Contact: latex allergy – Latex protein – itchy rash, redness, blisters and scaling (dermatitis) – National Taiwan University Hospital 6.8% in 1997 – Hospitals in central Taiwan 8.25% in 1998 Japanese Cedar Ragweed 17 Eukaryotes Higher Animals Sources: – pets or laboratory animals – rats, rabbits, cats, dogs, monkeys, etc. Exposure routes/modes – animal bites – danders – pets infested with arthropods Zoonoses 18 Eukaryotes Arthropods Arthropods Lice Tick Hosts Humans, Humans, dogs, dogs, etc. etc. Fleas Lice Cockroaches Humans, rats, cats, etc. Transmission bites/stings mode Tick Mites feces, body fragment Fleas Mites Cockroaches 19 Droplet Settling and Retention Time The time between droplets generated by a patient and settling onto the floor (20C,1atm) Diameter of droplets (μm) Time Settling Velocity (cm/s) 10 7~8 min 0.3 5 20~25 min 0.075 1 ~14 Hr 0.003 0.1 ~20 day 0.000087 20 Droplet Size and Retention Time Evaporative time for different sizes of droplets (the influence of temperature is insignificant) - RH: 50% Diameter of droplets (μm) Evaporative Time (sec) 10 0.15 5 0.02 1 0.0017 Diameter of droplets (μm) Evaporative Time (sec) 10 0.375 5 0.05 1 0.0043 - RH: 80% 21 1.2 Definition of Biohazards Biological hazards or Biohazards Biological hazards or biohazards are all of the forms of life (as well as the nonliving products they produce) that can cause adverse health effects. – These hazards are plants, insects, rodents, and other animals, fungi, bacteria, viruses, and a wide variety of toxins and allergens. (Yassi et al., 2001) 22 Unit 2 Sources and Modes of Transmission 2.1 Sources of Biohazards 1.Host: Infected humans or animals Human: tuberculosis, influenza, enterovirus infection Animals: rabies, mad cow disease 2. Environmental pathogens pollens fungi Legionella in contaminated water 24 Routes of Entry Inhalation (respiratory tract) tuberculosis, influenza, measles Ingestion (GI tract) contaminated food: Typhoid, cholera, Hepatitis A Through skin or mucous membranes Schistosomiasis, trachoma, bloodborne infectious diseases 25 2.2 Modes of Transmission 1a.HostInhalation Direct transmission Indirect transmission droplets e.g., influenza, enterovirus infection, meningococcal meningitis Inhalation Airborne (droplet nuclei) e.g., tuberculosis, measles 26 2.2 Modes of Transmission 1b.HostIngestion kiss Direct transmission vehicle Food: enterovirus, Hepatitis A Contaminated objects: clothes, bedding Water: Typhoid, cholera Ingestion Indirect transmission • Mechanical transmission pathogens are carried to the site of infection, e.g., flies vector • Biological vector-borne transmission pathogens need to grow and multiply in vectors, e.g., mosquitoes 27 2.2 Modes of Transmission 1c.HostSkin, Mucous membrane sex, vertical transmission, bites/stings, physical contact Direct transmission e.g., Syphilis, AIDS, Athlete's foot, Hepatitis B sharps (needles, scalpels): Indirect transmission Skin/ Mucous membranes Body fluid/ Blood e.g., Hepatitis B transfusion: e.g., Hepatitis B, AIDS vector (mosquitoes): e.g., Dengue fever, malaria 28 2.2 Modes of Transmission 2. Environmental pathogens fungi pollen protozoa bacteria others Air e.g., hay fever, legionnaire’s disease Vehicle (contaminated water or food) e.g., aflatoxin poisoning, cholera penetration, wounds, physical contact e.g., schistosomiasis, tetanus Inhalation Ingestion Skin/ Mucous membranes 29 Unit 3 Adverse Effects Adverse Effects of Biohazards Infection: growth and multiplication of an infectious agent in the body (e.g., influenza, measles, tuberculosis) Allergy: repeated exposure to antigens, which stimulate a specific immunological response (e.g., allergic pneumonia, asthma, allergic rhinitis) Toxicosis : resulting from exposure to toxins produced by organisms such as bacterial endotoxin and exotoxin, mycotoxin, etc. (e.g., fever, chill, impairment of lung function) Others: (e.g., psychological effects) 31 Fir Tree Found Growing in Russian Man’s Lung Surgeons find fir tree 'growing inside patient's lung' Russian surgeons have claimed to have found a two-inch fir tree growing inside a man's lung. The amazing 'discovery' was apparently made when they opened up Artyom Sidorkin, 28, to remove what they thought was a serious tumor. http://www.telegraph.co.uk/news/worldnews/europ e/russia/5152953/Surgeons-find-fir-tree-growinginside-patients-lung.html 32 Psychological effects 33 Unit 4 Biohazard Prevention and Control 4.1 Prevention Classification of Infectious Microorganisms by Risk Group – based on National Science Council Guidelines for Experiments involving Genetic Recombination 2004(行政 院國家科學委員會93年6月增修版之「基因重組實驗守 則」) – Other references: NIH Guidelines for Research involving Recombinant DNA Molecules 2011; U.S. Department of Health and Human Services Biosafety in Microbiological and Biomedical Laboratories 2009 (5h Ed); WHO Laboratory Biosafety Manual 2004 (3rd Ed). 35 Risk Group 1 (RG1) Agents Agents not associated with disease in healthy adult humans Examples 1. Viruses – adeno-associated virus (AAV) types 1 through 4; and recombinant AAV constructs, in which the transgene does not encode either a potentially tumorigenic gene product or a toxin molecule and are produced in the absence of a helper virus. 2. Bacterial Agents – asporogenic Bacillus subtilis or Bacillus licheniformis – nonpathogenic Escherichia coli (Escherichia coli K-12) 36 Risk Group 2 (RG2) Agents Agents associated with human disease that is rarely serious and for which preventive or therapeutic interventions are often available Examples: 1.Viruses – Dengue virus serotypes 1, 2, 3 and 4 – Hepatitis A, B, C, D, and E viruses – Measles virus – Mumps virus – Coxsackie viruses types A and B 37 Risk Group 2 (RG2) Agents 2. Bacterial Agents including Chlamydia – Escherichia coli - all enteropathogenic, enterotoxigenic, enteroinvasive and strains bearing K1 antigen, including E. coli O157:H7 – Helicobacter pylori 3. Fungal Agents – Cryptococcus neoformans – Penicillium marneffei 4. Parasitic Agents – Ancylostoma human hookworms – Heterophyes – Enterobius 38 Risk group 3 (RG3) Agents Agents associated with serious or lethal human disease for which preventive or therapeutic interventions may be available Examples: 1. Viruses and Prions – Transmissible spongioform encephalopathies (TME) agents (Creutzfeldt-Jacob disease and kuru agents) – Hantaviruses – Human immunodeficiency virus (HIV) types 1 and 2 – SARS-associated coronavirus (SARS-CoV) 39 Risk Group 3 (RG3) Agents 2. Bacterial Agents including Rickettsia – Mycobacterium tuberculosis – Coxiella burnetii 3. Fungal Agents – Coccidioides immitis – Histoplasma capsulatum – H. capsulatum var. duboisii 40 Risk Group 4 (RG4) Agents Agents likely to cause serious or lethal human disease for which preventive or therapeutic interventions are not usually available Examples: Viruses – – – – Arenaviruses : e.g., Lassa virus Bunyaviruses: Crimean-Congo hemorrhagic fever virus Filoviruses : Ebola virus, Marburg virus Flaviruses (Group B Arboviruses): Tick-borne encephalitis virus complex – Herpesviruses (alpha) – Paramyxoviruses : Equine morbillivirus 41 Relation of Risk Groups to Biosafety Levels, Practices and Equipment Source: WHO 2004; Laboratory Biosafety Manual, p2. 42 Pathogen Safety Data Sheets and Risk Assessment http://www.phac-aspc.gc.ca/lab-bio/res/psds-ftss/index-eng.php Section I: Infectious Agent – Name – Synonym or Cross Reference – Characteristics Section II: Hazard Identification – Pathogenicity/Toxicity – Epidemiology – Host Range – Infectious Dose – Mode of Transmission – Incubation Period – Communicability Section III: Dissemination – Reservoir – Zoonosis – Vectors Section IV: Stability and Viability — Drug susceptibility — Drug resistance — Susceptibility to disinfectants — Physical inactivation — Survival outside host Section VII: Exposure Controls /Personal Protection – Risk group classification – Containment requirements – Protective clothing – Other precautions Section V: First Aid /Medical — Surveillance — First Aid/Treatment — Immunization — Prophylaxis Section VIII: Handling and Storage – Spills – Disposal – Storage Section VI: Laboratory Hazards — Laboratory-acquired infections — Sources/Specimens — Primary Hazards — Special Hazards Section IX: Regulatory and Other Information – Regulatory information – Updated – Prepared by 43 General Prevention Methods – Environmental Management – Personnel Management 45 Environmental Management Source removal (most important prevention method) Regular and proper environmental cleaning and disinfection Humidity control Air quality management 46 Air Quality Management Mechanical ventilation systems – General ventilation: airflow type and direction – Local exhaust ventilation: Biosafety cabinet Air cleaning devices – High-efficiency particulate air (HEPA) filter – Germicidal devices Negative-pressure environments – Negative pressure isolation room – Biosafety Level 3 (BSL-3) Laboratory 47 Personnel Management Personal hygiene (e.g., wash hands) Personal health management (e.g., immunization) Standard microbiological practices Personal protective equipment (the last resort for prevention) -wear lab coats -wear gloves and masks 48 Hand Hygiene Technique with Soap and Water Duration of the entire procedure: 40-60 seconds 49 Source: WHO 2009; WHO Guidelines on Hand Hygiene in Health Care, p156. 50 Standard Microbiological Practices Restrict or limit access when working Wash hands before and after work Decontaminate work surfaces before and after work, and immediately after spill Minimize splashes and aerosols Prohibit eating, drinking, smoking, handling contact lenses and applying cosmetics in the laboratory Prohibit mouth pipetting Use durable, leak proof containers to transport infectious material Decontaminate infectious waste, including specimens and cultures, before disposal Handle sharps with care; place sharps in puncture-resistant containers used for disposal or transport 51 Minimize splashes and aerosols Any laboratory manipulation can produce aerosols. Open top test tubes or containers are of particular concern. Measures can be taken to minimize these hazards. Source: WHO & DIH, http://www.apbtn.org/apbtn/trainingMaterials.html 52 How to Remove Gloves (1) 1.Pinch one glove at the wrist level to remove it, without touching the skin of the forearms, and peel away from the hand, thus allowing the glove to turn inside out 2.Hold the removed glove in the gloved hand and slide the fingers of the ungloved hand inside between the glove and the wrist. Remove the second glove by rolling it down the hand and fold into the first glove. 3. Discard the removed gloves http://whqlibdoc.who.int/publications/2009/9789241597906_eng.pdf 53 How to Remove Gloves (2) Key points: 1. Do not touch the outside of the gloves with bare hands during the process: gloves to glove and hand to hands only 2. If the gloves are very dirty, rinse them with water before removing. 3. Removed gloves must be discarded in a biohazard bag or receptacle. 54 4.2 Control Disinfection and Sterilization – Definitions: • Disinfection: A physical or chemical means of killing microorganisms, but not necessarily spores. • Sterilization: A process that kills and/or removes all classes of microorganisms and spores. – Cleaning prior to disinfection and sterilization: • Dirt, soil and organic matter can shield microorganisms and can interfere with the killing action of decontaminants (antiseptics, chemical germicides and disinfectants). • Precleaning (remove organic compounds) is essential to achieve proper disinfection or sterilization. 55 Chemical germicides(1) Alcohols Principle: – 75% alcohol kills pathogens by denaturing their proteins (coagulate proteins) and has better efficiency of disinfection. – 95% alcohol makes pathogens form a protective film ,which prevents any alcohol from entering the cell, decreasing disinfection efficiency. Notice: – Alcohols are active against vegetative bacteria, fungi and lipidcontaining viruses but not against spores. Their action on nonlipid-containing viruses (e.g., Enterovirus) is variable. – Alcohols tend to swell and harden rubber and certain plastic tubing after prolonged and repeated use, as well as bleach rubber and plastic tiles. – Simple preparation for 75% alcohol: 95% alcohol: water = 3:1 → approximately 71.25% 56 Chemical germicides(2) Chlorine (sodium hypochlorite) Principle: – Oxidation Notice: – A 1:100 dilution of 5% sodium hypochlorite is recommended; i.e. for household bleach containing 5% sodium hypochlorite, use 1 part bleach to 99 parts cold tap water (1:100 dilution) for disinfecting surfaces. – Bleach may cause irritation to mucous membranes, skin and respiratory tract; decomposes under light and heat; and reacts easily with other chemicals to produced toxic gas. 57 Physical agents(1) Ultraviolet Radiation Principle: – UV radiation at a wave-length of 254 nm causes alterations/defects in DNA, which inhibits pathogen replication. Notice: – UV light lacks penetrating power. Microorganisms beneath dust particles or beneath the work surface are not affected by UV radiation. – Require at least 20 minutes of exposure (contact time), but it is ineffective in spores and endospores. – UV rays are harmful to human cells. 58 Physical agents(2) Autoclaving Principle: – Applying high-pressure steam (wet heat) to permeate materials within the autoclave: complete sterilization by coagulation and denaturation of proteins Notice: – Used for items (including containers) resistant to heat and moisture – For most purposes, the following cycles will ensure sterilization of correctly loaded autoclaves: (1) Sterilizing 3 min at 134 °C (2) Sterilizing 10 min at 126 °C (3) Sterilizing 15 min at 121 °C (4) Sterilizing 25 min at 115 °C – Biomedical waste can be treated as general industrial waste, given that the waste has been autoclaved as regulated by the Dept. of Health, Executive Yuan. 59 Physical agents(3) Photocatalyst Principle: – When the photocatalyst (TiO2) is illuminated by UV radiation, electron/hole pairs are produced on the TiO2 surface, generating free radicals to decompose pollutants. Notice: – UV radiation with a wavelength of 365nm is needed to excite the titania catalyst, producing free radicals to facilitate redox reactions. – In order to inactivate pathogens, direct contact with the TiO2 surface for at least 30 minutes is required. – Human body and eyes should not be directly exposed to UV light. 60 Biological Safety Cabinet 61 Biosafety Cabinet (BSC) vs. Laminar Flow Clean Bench BSC – Use clean negative-pressure laminar flow to prevent emission of contaminated air – Main purpose: protect the operator, the laboratory environment and work materials from exposure to infectious agents Class II A2 BSC classifications based on the protection efficiency – Class I: less common – Class II: commonly used A. Certification Requirement: NSF/ANSI 49-2007(US Standard )/BS EN 12469:2000(European Standard ) B. Types: A1, A2(B3 originally), B1, B2 – Class III: provides the highest level of personnel protection and is used for Risk Group 4 agents. Positive-pressure laminar flow clean bench: provide product protection only 62 Positive-pressure Laminar Flow Clean Bench ≠ Biosafety Cabinet Exhausted outward to operator, providing product protection only Need to shield glass window (usually use curtain fabrics) while UV light is on. Replace the top priminary filter regularly (e.g., every 500 hrs) Air flow 63 Using Class II Biosafety Cabinet Open flames should be avoided: create turbulence in the airflow, compromising protection of both the worker and the work Do not block the front or rear grille openings: create turbulence in the airflow, disrupting laminar flow Ensure that the sash is set in the correct operating position (about 20 cm above the base of the opening ) The top opening is the filtered air outlet which should be cleaned regularly and cannot be blocked. Close the sash while the UV light is on. The primary filter of Type A1 BSC can be changed by lab personnel regularly HEPA filter should be replaced by a qualified professional regularly. Annual certification is required. 64 Temporary Storage and Management of Biomedical Waste (blood, cell and tissue samples, animals) 65 The Definition of Biomedical Waste Healthcare waste: WHO – The total waste stream from a healthcare facility, including the waste generated in the diagnosis, treatment, prevention, rehabilitation and related research activities. Biomedical waste: Taiwan – The waste generated during the process of medical treatment/test, autopsy, quarantine, research and production of drugs or biomaterials at healthcare facilities, clinical/medical laboratories, Biosafety Level 2 and above laboratories of industrial or research institutes, genetics/biotechnology laboratories and biotechnology/pharmaceutical factories. – including genotoxic waste, sharps, infectious waste 66 The Disposal Procedures of Biomedical Waste Three major steps: storage, transport and disposal Collect, segregate, pack and label the waste before storage. The new and old labels of biomedical waste Biomedical Waste Infectious Industrial Waste (announced on 2007.5.11) (abolished) 67 Segregation and Storage of Biomedical Waste Combustible Medical Waste – Seal and store in a red combustible container and label as biomedical waste; can be stored up to 1 day at room temperature, 7 days at 5 C, and 30 days at 0 C. Non-Combustible Medical Waste – Seal and store in a yellow unpenetrable container and label as biomedical waste; need sterilization. (Date and temperature of storage and the sign of biomedical waste should be lablled on the container.) 68 Cardboard containers have to be sealed; plastic bags have to be tied. Proper label and symbol in Chinese Sturdy Leak proof For incineration: red container For sterilization: yellow container Only for infectious waste, not for sharp devices Figure 2-15 The storage containers for infectious waste Source: Taiwan EPA 2008; Handbook of Medical Waste Management. 69 with a fitted cover Proper label and symbol in Chinese Sturdy and punctureresistant (made of metal or rigid plastic) Leak-proof Sharps containers Figure 2-14 The containers for sharp waste Source: Taiwan EPA 2008; Handbook of Medical Waste Management. 70 Combustible Waste Use leak-proof containers fitted with covers NonCombustible Waste Discard disposable sharps (e.g., needles and scalpel blades) into puncture-proof (made of metal or high-density plastic ) and rigid containers fitted with covers Disinfect or autoclave contaminated (with Waste containing infectious agents can be treated potential infectivity) reusable materials as industrial waste after sterilization (e.g., gloves, (e.g., glass pipettes, serum bottles) disposable centrifuge tubes, blood collection tube and etc.) 71 Contaminated (with potential infectivity) materials that can be reused after sterilization or autoclaving inappropriate 0.5% bleach Any contaminated (with potential infectivity) reusable materials should not be cleaned prior to autoclaving/sterilization. Any necessary washing or repair should be done after autoclaving/sterilization. Vertical soak (not completely immersed) Horizontal soak (aspirate disinfectant before removing the pipette dispenser, and then immerse the whole pipette in the disinfection box) Examples: 1. Pipettes used for RG1 and general cell culture should be immersed in the disinfectant first and then washed. 2. Reusable materials (e.g., glass pipettes, serum bottles) used to handle RG2 and above infectious materials can be soaked in the disinfectant first, autoclaved, and then washed. 72 Temporary Storage and Disposal of Animal Carcass Install sufficient refrigerators in animal rooms to store animal carcasses temporarily. After removing from the freezer, thaw the carcass before incineration to prevent incomplete combustion and waste of fuel; defrost the carcass in an appropriate place to prevent contamination by melting water (use a leak-proof container). Send innocuous animal carcasses directly to landfill or incineration; infectious animal carcasses should be packaged in the biohazard labeled plastic bags, autoclaved and then treated as innocuous animal carcasses. Radioactive animal carcasses should be packaged in the plastic radioactive waste bags and dried under 60-70C using a specialized oven; using a microwave to heat and dehydrate animal carcasses can prevent odor generated during the process; dehydrated animal carcasses can then be treated as radioactive waste. 73 Contaminated biological materials that can be incinerated directly or incinerated after sterilization (e.g., organs or carcasses of innocuous or infectious animals) 74 Current Laboratory Waste Disposal in Academic Institutes in Taiwan Waste Control, Decontamination Without BSC (BSL1) and Management (N=121) Proper segregation and storage of infectious waste Sterilization of infectious waste before disposal With BSC (BSL2) (N=161) 74 61.2% 136 84.5% 103 85.1% 151 93.8% Source: Taiwan IOSH 2006; Investigation of Facility and Installation at Biosafety-Level-2 Laboratories (IOSH95-H101). 75 Current Laboratory Waste Disposal in Academic Institutes in Taiwan (cont.) Not using leak proof container and uncovered Not using leak proof container and uncovered Source: Taiwan IOSH 2006; Investigation of Facility and Installation at Biosafety-Level-2 Laboratories (IOSH95-H101). Not using the biomedical waste bag resistant to high temperature and pressure 76