20BT2057 Bioethics, Ipr And Biosafety PDF

L T P C 20BT2057 BIOETHICS, IPR AND BIOSAFETY 3 0 0 3 Course Teacher Dr. R.S. David Paul Raj PhD Associate Professor Department of Biotec...

L T P C 20BT2057 BIOETHICS, IPR AND BIOSAFETY 3 0 0 3 Course Teacher Dr. R.S. David Paul Raj PhD Associate Professor Department of Biotechnology School of Agriculture and Biosciences Karunya Institute of Technology and Sciences 1 2 3 Biosafety is defined by the World Health Organization (WHO): Biosafety comprises “the containment principles, technologies and practices that are implemented to prevent unintentional exposure to pathogens and toxins or their accidental release”. Biosafety is defined as the application of a combination of laboratory practices and procedures, laboratory facilities, and safety equipment when working with potentially infectious micro-organisms to protect the laboratory staff and, through them, the general public. 4 The events of September 11, 2001, and the anthrax attacks in October of that year reshaped and changed, forever, the way we manage and conduct work in biological and clinical laboratories. 5 WHO has designated five Collaborating Centres on Biosafety in the recent past. Of these, two are in the USA and one each in Canada, Australia and Sweden. Recognizing the importance of the subject and the need for platforms for professionals to share views and to advise national authorities, several regional bodies of professionals have been established. These include the American Biosafety Association, the Asia-Pacific Biosafety Association and the European Biosafety Association. 6 BRIEF HISTORY OF BIOSAFETY Innovation and development of biosafety in the United States is reflected accurately in the history and pre-history of the American Biological Safety Association (ABSA). The first unofficial meeting was held on April 18, 1955 at Camp Detrick (now Fort Detrick) and involved members of the military. “The Role of Safety in the Biological Warfare Effort.” Beginning in 1957. There were striking changes in the meetings in 1964-1965: the NIH and CDC joined for the first time, along with a number of other relevant federal Agencies. By 1966, the attendees included universities, private laboratories, hospitals, and industry. 7 In 1974, the United States Postal Service and Department of Transportation introduced regulations for shipping of etiologic agents (microorganisms and toxins that cause disease in humans). New safety programs and trainings were introduced. The designation of 4 levels of biosafety originated in the mid-1970s, and the safety requirements for research with recombinant DNA were hotly debated. The primary agencies involved are the Department of Labor (DOL), Occupational Safety and Health Administration (OSHA), Health and Human Services (HHS) Centers for Disease Control and Prevention (CDC), and the U.S. Department of Agriculture (USDA) Animal and Plant Health Inspection Service (APHIS). 8 9 10 11 Biotechnology has wide application in the field of healthcare, industry, agriculture and environment. The industrial applications include fermentation based products, biotech instruments, equipment’s, bio-fuel, bioenergy and bio-mining. In case of medicine it used in vaccines, diagnostics, monoclonal antibodies, stem cells and tissue specific delivery system. In the field of agricultural, the application includes biofertilizer, bio- pesticides, GM-planting materials and hybrids, micro-propagation and animal improvement. In case of environment, the biotechnolocal application plays a crucial role in soil and water remediation. 12 In mid 1980s the American NIH guidelines are developed the rules for laboratory work associated with genetic engineering techniques on the basis of national and international laws of different countries. The first international biosafety guideline was framed in the year 1986 by OECD (Organisation for Economic Co-operation and Development) also known as blue book. This guideline is mainly for the use of recombinant DNA organisms in various sectors like agriculture, industry and environment. 13 Principles of risk assessment Risk assessment is backbone of biosafety and effective tool to determine the proper biosafety level and safety practices for the work considering health issues, environmental, ecological issues and social issues. Types of Risk Assessment Risk Assessments: Microbiological : Risk assessment in Microbiological aspects consists of virulence, infection route, mode of transmission, survival in the environment, infectious dose, effective preventative and therapeutic treatments method availability, impact of introduction and/or release into the environment or the public and concentration of the pathogen. 14 Risk Assessments: Animal Biosafety Risk assessments in model animals are following: i. Only known permissive species should be used ii. Nature of animal to be known including animal’s aggressiveness, bite nature and tendency to scratch. iii. Principal investigator must be identified for its risks, exposure route, symptom of infection, risk on personnel and immunizations iv. Suitable surveillance programme for the staff along with safety operation manual must be adopted. v. Display of necessary biohazard warning vi. Suitable medical surveillance programme for the staff must be adopted and safety operations manual must be used. vii. Only experimental animals must be maintained. viii. Biological safety cabinets (Classes I or II) and individual cages with suitable air supply with HEPA-filtered exhaust air must be provided. 15 Risk Assessments of Genetically Modified Plants Risk assessment of GM plants begins with hazard identification, hazard characterization, exposure characterization, risk characterization, risk management strategies, and an overall risk evaluation. Risk assessment of GMOs considers seven specific areas of concern such as persistence and invasiveness nature of GM plant. This also includes plant to plant gene transfer. Gene transfer between pant-to-micro-organism Interaction of the GM plant with target organisms Interaction among GM plant with non-target organisms, selection of suitable and relevant functional group for risk assessment including impact of cultivation, management and harvesting methods, production and receiving environment. 16 Risk assessments of genetically modified organisms (GMOs) It is important to assess the risk associated with GMOs before release into environment as well as impact on human. Each individual case needs to be assessed for pre-marketing safety assessment following the structured and integrated approach. It includes hazard identification, risk characterization and exposure assessment. 17 Risk Management In risk management the main three components are impact assessment, public awareness/participation, and the design of regulatory systems. The protocol establishes to measures strategies to regulate, manage, and control risks identified in the provision of risk assessment. The national regulatory frame work to ensure the bio-safety decision according to the scientific risk assessment. For commercial release of GMOs, it needs to be studied involving a higher number of GMOs under complex ecosystems over a time at different sites to reveal the interaction among species and ecosystem. 18 Regulations and Guidelines on Biosafety of Recombinant DNA Research & Biocontainment 2017 Containment Containment encompasses safe methods (Combination of facilities, practices and procedures) for managing risk-inherent microorganisms, GE organisms or cells in the laboratory environment where they are being handled or maintained. Selection of appropriate containment strategy will ensure safety to laboratory workers, outside people and the environment from hazardous microorganisms, GE organisms or cells by: i. Reducing the exposure, and ii. Preventing their escape and establishment in a natural environment. Principle The principle is the protection of all identified elements from risk(s) posed by organisms (includes risk-inherent; GE and non-GE microorganisms, animal, plants, arthropods, aquatic animals, etc) during their use in laboratory. In practice, it should be achieved in realization of three interrelated steps: i. Identification of elements that should be protected: Containment measures should ensure protection of laboratory worker(s) (Primary elements) who have maximum possibility of exposure to the organisms. In addition, the containment measure should also prevent the escape of organisms and so ensure protection of persons outside the laboratory and the environment (Secondary elements). ii. Identification of potential risk(s) associated with organism(s): It involves assessment of risk(s) associated with the organisms and their classification to appropriate risk groups based on: a. Pathogenicity of the organism towards humans/animals/plants. b. Modes of transmission and host range of the organism. c. Availability of effective preventive treatments or curative medicines. d. Capability to cause epidemics. Physical Containment The strategy is to physically confine the organism under study that can be feasibly adopted to prevent or minimize its exposure to worker and environment ensuring the risk(s) can be prevented or mitigated. It is achieved through the use of three elements of containment i.e. Procedures, Safety equipment(s) and Facility design(s). The protection of personnel(s) and the immediate laboratory environment from exposure to organisms (includes risk-inherent; GE and non-GE microorganisms, animals, plants, arthropods, aquatic animals, etc.) is provided by ‘Procedures’ and the use of appropriate ‘Safety equipment(s)’ (Primary containment). The protection of the environment external to the laboratory from exposure to risk-inherent materials is provided by a combination of ‘facility design’ and operational practices (Secondary containment). The elements are not in hierarchy and should be used with equal priorities in combination to ensure a successful containment  Risk Group 2 Pathogenic for humans Unlikely a serious hazard Treatment and preventive measures available Limited risk of spread of infection CDC, Yersinia pestis laboratory  Risk Group 3 Pathogenic, cause serious disease Effective treatment and preventive measures usually available Little person-to-person spread Laboratory in Lyon France  Risk Group 4 Lethal, pathogenic agent Readily transmittable – direct, indirect Effective treatment and preventive measures not usually available National Institute for Infectious Diseases, Rome, Italy Procedure These must be followed by workers involved in research and handling of organism in consideration of: i. Strict adherence to standard microbiological practices and techniques. ii. Awareness of potential hazards. iii. Providing/arranging for appropriate training of personnel. iv. Selection of safety practices in addition to standard laboratory practices if required. It is emphasized that good laboratory practice is fundamental to laboratory safety and cannot be replaced by any other means, which can only supplement it. Safety Equipments Any equipment that contributes to personnel protection either directly or indirectly from the hazardous biological material is considered for containment. It includes: i. Instruments like biological safety cabinets, autoclave and a variety of enclosed containers (e.g. safety centrifuge cup). ii. The biological safety cabinet (BSC) is one of the principal devices used to provide workers safety from hazardous microorganisms and infectious aerosols. iii. Three types of BSCs (Class I, II, and III) are used in biosafety level facilities. Safety and functionality of each instrument must be monitored monthly for effectiveness and calibrated annually before commencing operations. iv. Equipment such as autoclaves and biological safety cabinets must be validated with appropriate methods (usually by a certified examiner) before being taken into use. The results of the monitoring and calibration must be documented. v. Recertification should take place at regular intervals, according to the manufacturer’s instructions. vi. If any equipment is found to be defective and the defect has not been corrected, the equipment must be clearly marked to show that it is defective and must not be used for any purpose until the defect has been corrected. Personal protective equipment (PPE) such as gloves, coats, gowns, shoe covers, boots, respirators, face shields and safety glasses, etc. The Head of the laboratory, after consultation with the biosafety officer and IBSC, should ensure that adequate equipment is provided and that it is being used properly. In selecting safe laboratory equipment, the general principles that should be considered include: i. Designed to limit or prevent contact between the operator and the infectious organisms. ii. Constructed of materials that are impermeable to liquids, corrosion-resistant and meet structural strength requirements. iii. Fabricated to be free of burrs and sharp edges. iv. Designed, constructed and installed to facilitate simple operation and to provide ease of maintenance, accessibility for cleaning, and ease of decontamination and certification testing. These are general principles. Detailed performance and construction specifications may be required to ensure that the equipment possess necessary safety features.  Microbiological Biosafety Level (BSL) Facilities Biosafety Level 1 (BSL-1): BSL-1 will be applicable for: i. Isolation, cultivation and storage of Risk Group (RG) 1 microorganisms those are abundant in natural environment. ii. Experiments on RG 1 microorganisms provided that the experiments will not increase environmental fitness and virulence of the microorganisms. iii. Category I genetic engineering experiments on microorganism: This category includes experiments which generally do not pose significant risk(s) to laboratory workers, community or the environment and the modifications have no effect on safety concerns. Examples are: a. Insertions of gene into RG 1 microorganism from any source, deletions, or rearrangements that have no adverse health, phenotypic or genotypic consequence. Modification should be well characterized and that the gene functions and effects are adequately understood to predict safety. b. Experiments involving approved host-vector systems (refer to Annexure 3) provided that the donor DNA is originated from RG 1 microorganism, not derived from pathogens. The DNA to be introduced should be characterized fully and will not increase host or vector virulence. c. Experiments involving the fusion of mammalian cells which generate a non-viable organism, for example, the construction of hybridomas to generate monoclonal antibodies. d. Any experiments involving microorganism belonging to RG 1. For e.g. self-cloning, fusion of protoplasts between non-pathogenic RG 1 organism. Biosafety Level 2 (BSL-2): BSL-2 will be applicable for: i. Isolation, cultivation and storage of RG 2 microorganisms. ii. Handling of environmental samples collected from environment that is unlikely to contain pathogens. Isolation of microorganisms from those samples and subsequent experiments. iii. Experiments on RG 2 microorganisms or isolates from environment mentioned above, provided that the experiments will not increase environmental fitness and virulence of the microorganisms. iv. Category II genetic engineering experiments on microorganism: These experiments may pose low-level risk(s) to laboratory workers, community or the environment. Examples are: a. Experiments involving the use of infectious or defective RG 2 viruses in the presence of helper virus. b. Work with non-approved host/vector systems where the host or vector either: does not cause disease in plants, humans or animals; and/ or is able to cause disease in plants, humans or animals but the introduced DNA is completely characterized and will not cause an increase in the virulence of the host or vector. experiments using replication defective viruses as host or vector. c. Experiments with approved host/vector systems, in which the gene inserted is: a pathogenic determinant; not fully characterized from microorganisms which are able to cause disease in humans, animals or plants; or an oncogene. d. Modification leading to persistent transient disruption of expression of gene(s) that are involved directly or indirectly in inducing pathogenicity, toxicity, survival, or fitness. Modification should be well characterized and the gene functions and effects are adequately understood to predict safety. e. Work involving fragments of Transmissible Spongiform Encephalopathy (TSEs) proteins or modified TSEs proteins that are not pathogenic and is not producing any harmful biological activity. f. Experiments in which DNA from RG 2 or 3 organisms are transferred into non-pathogenic prokaryotes or lower eukaryotes. However, handling of live cultures of RG 3 organism should be performed in BSL-3 laboratory. Biosafety Level 3 (BSL-3): BSL-3 will be applicable for: i. Isolation, cultivation and storage of RG 3 microorganisms. ii. Handling of environmental samples collected from environment that is likely to contain pathogens of potential disease consequences. Isolation of microorganisms from those samples and subsequent experiments. iii. Experiments on RG 3 microorganisms or isolates from environment mentioned above provided that the experiments will not increase environmental fitness and virulence of the microorganisms. iv. Category III and above genetic engineering experiments on microorganism: These kinds of experiments pose moderate to high risk(s) to laboratory workers, community or the environment. Examples are: a. Experiments on RG 2 and RG 3 microorganisms where insertion of gene directly involved in production of toxin or allergen or antimicrobial compounds. b. Insertions of gene into RG 3 microorganisms from any source, deletions, or rearrangements that affect the expression of genes, whose functions or effects are not sufficiently understood to determine with reasonable certainty if the engineered organism poses greater risk(s) than the parental organism. c. Insertions of nucleic acid from any source, deletions, or rearrangements that have known or predictable phenotypic or genotypic consequence in the accessible environment that are likely to result in additional adverse effects on human and/or animal health or on managed or natural ecosystems, e.g., those which result in the production of certain toxins. d. Research involving the introduction of nucleic acids (recombinant or synthetic) into RG 3 organisms or organisms listed in SCOMET items (http://dgft.gov.in). e. Genetic engineering of organisms isolated from environment where there are reported cases of disease prevalence and possibility of presence of infectious microorganisms Biosafety Level 4 (BSL-4): BSL-4 laboratory is the maximum containment laboratory. Strict training, strictly restricted access and supervision are required and the work must be done under stringent safety conditions and positive pressure personnel suits. BSL-4 will be suitable for: i. Isolation, cultivation and storage of RG 4 microorganisms. ii. Handling of samples collected from environment/patients that are likely infected with RG 4 organisms with serious/fatal health effects. iii. Experiments on RG 4 microorganisms or isolates from environment/patients mentioned above to find remedial measures. iv. Category III and above genetic engineering experiments on microorganisms involving introduction of nucleic acids (recombinant or synthetic) into RG 4 microorganisms or exotic agents. 45