1. Biological Risk: Basic Concepts and Classifications Jebunnesa Chowdhury

2.  Biological Risk Biological risk refers to naturally occurring or human made risk caused by exposure to biological agent or microorganism. The following terminologies associated with biological risks are defined or described:  Risk - likelihood that under particular conditions of exposure an intrinsic hazard will represent a threat or harm; risk is a function of hazard and exposure.  Risk = hazard x exposure Or Risk = hazard x probability x consequence  Hazard – intrinsic (it is there or it is not) potential of a material to cause harm to human health and/or the environment; also synonymous to threat  Exposure – the extent and the duration of or the frequency with which the operator is exposed to the hazard  Likelihood – probability of something happening  Consequence – adverse effects or outcome caused by an event  Stakeholders – individual, groups and institutions who may affect, be affected by, or perceive themselves to be affected by the decision, activity or risk; synonymous with ‘interested parties

3.  Biological agent - a micro-organism, cell culture, or human endoparasite, whether or not genetically modified, and products derived from them which may cause infection, allergy, toxicity or otherwise create a hazard to human health and the environment; also synonymous with biohazard  Microorganisms - a microbiological entity, cellular or non-cellular, which is capable of replication or of transferring genetic material

4. Naturally Occurring vs. Biotechnological Risks Biological risk can be classified into two broad categories: naturally occurring or human- caused. Naturally occurring biological risks – includes-  (1) the emergence of antibiotic resistant bacterial infections (tuberculosis, pneumonia, flu epidemic);  (2) naturally emerging pathogens attributed to deforestation (monkey pox, Ebola, Lassa fever);  (3) spreading of a zoonosis i.e. infected animal population conveying the disease to humans via direct contact, vector or water/foodstuffs;  (4) toxins arising from certain molds and fungi (deoxynivalenol, alflatoxins, ochratoxin a);  (5) parasitic infection outbreaks in humans;  (6) invasive alien species (plants, animals and microorganism)

5. Human caused or related biological risks – can be further classified into: (1) deliberately induced risks such as the use of harmful biological agents through warfare or terrorism; (2) biotechnological risks such as products of traditional cross breeding and selection, mutation and modern biotechnology.

6. Biological Agents and Risk Groups Laboratory associated infections (LAI) has been documented since the beginning of the 20th century. The historical accounts of LAIs, though relatively infrequent and the advent of modern biotechnology raised awareness about the hazards of infectious microorganisms and the risks these posed to laboratory workers who handle them and the community if they escaped from the laboratory.  There are three ways that will bring workers into contact with materials that may pose a biological risk. These are: 1. exposure as a result of working with biological agents – areas of work include a microbiology laboratory; greenhouse, animal house; activities include isolation, identification and culture of microorganisms or cells including materials used for genetic modification, intentional contact with animals and plants and materials that originate from animals and plants as part of planned experimental work.

7. Biological Agents and Risk Groups  2. exposure which does not result from the work itself but is incidental to it, mainly because biological agents are present as contaminants - areas and activities include farming, refuse collection, sewage treatment, handling human body fluids and excreta; handling materials that may be contaminated by these materials such as hypodermic needles or sewage treatment plant.  3. exposure which is not a result of the work that you do – unintentional contact with animals and animal and plant materials or people in the workplace.

8. The World Health Organization (WHO) has recommended an agent risk group classification for laboratory use that describes four general risk groups based on the risk criteria/factors described below.  Pathogenicity of the agent or its product - inherent risks of a pathogen are based on factors such as the severity of the disease it causes, its virulence and infectivity; ‘disease’ caused by agent’s products include toxicity, allergenicity, physiological activity (e.g. anti-nutritional).  Mode of transmission and host range of the agent – these are influenced by existing levels of immunity, density and movement of the host population, presence of appropriate vectors and standards of environmental hygiene.

9.  Availability of effective preventive measures - measures may include: prophylaxis by vaccination or antisera; sanitary measures, e.g., food and water hygiene; the control of animal reservoirs or arthropod vectors; the movement of people or animals; and the importation of infected animals or animal products.  Availability of effective treatment - includes passive immunization and post-exposure vaccination, antibiotics, and chemotherapeutic agents, taking into consideration the possibility of emergence of resistant strains.

10. Other considerations that maybe taken to account in classifying biological agents include  Origin/source – indigenous (native, local) or exotic (foreign, alien); e.g. exotic agents posed higher risks to human health because it may cause more severe infection with no available treatment  Ability of the organism to survive – dormancy or resting period; duration  Number/concentration of microorganism – the higher the number the greater the possibility of infection  Nature and route of transmission – inhalation (dust, aerosol), ingestion (food, drink, saliva), contact (cuts, bites, injection)

11.  The National Institute of Health, USA (NIH) Guidelines established a comparable classification of genetically modified hazard agents into a particular risk group using the same criteria indicated above. Many countries, including Bangladesh (see Annex 1 of the Bangladesh Biosafety Guidelines 2006), have adopted the WHO and NIH risk group classifications and criteria. The descriptions of the WHO and NIH risk groups are presented in Table 1 below.

13. C ontainment and Biosafety Levels  Containment refers to the ability to reduce or eliminate exposure of workers, other persons, and the outside environment to potentially hazardous agents. It is also used to describe safe methods, facilities and equipment for managing infectious materials in the environment where they are being handled or maintained. For experimental plants and animals and microorganisms that are restricted within a chosen outdoor environmental zone of control, the term confinement has been used.

14. The fundamental elements of containment include the following: 1. Safe laboratory practices and techniques- the most important element of containment; It refers to strict adherence to standard microbiological practices and techniques which require properly trained personnel, a biosafety or operations manual that identifies the hazards and specifies practices and procedures designed to minimize or eliminate exposures to the hazards. 2. Safety equipment (primary physical barrier or personal protective equipment) – primary barrier includes biological safety cabinets (BSCs), enclosed containers (e.g. centrifuge cups), and other engineering controls designed to remove or minimize exposures to hazardous biological materials. The BSC is the principal device used to provide containment of infectious splashes or aerosols generated by many microbiological procedures. Safety equipment also may include items for personal protection, such as gloves, coats, gowns, shoe covers, boots, respirators, face shields, safety glasses, or goggles.

15. The fundamental elements of containment include the following: 3. Facility design and construction (secondary physical barrier) - provides a barrier to protect persons outside the laboratory, and protects persons or animals in the community from infectious agents that may be accidentally released from the laboratory. The recommended secondary barrier(s) will depend on the risk of transmission of specific agents. Special design features include specialized ventilation systems to ensure directional air flow, air treatment systems to decontaminate or remove agents from exhaust air, controlled access zones, airlocks as laboratory entrances, or separate buildings or modules to isolate the laboratory.

16. 4. Biological barriers - natural barriers that limit either: (i) the infectivity of a vector or vehicle (plasmid or virus) for specific hosts, or (ii) its dissemination and survival in the environment; also include design of facilities and special practices for limiting or excluding the unwanted establishment, transfer of genetic information, and dissemination of organisms beyond the intended location; used for experiments involving recombinant DNA technology to replace hazardous agents and decrease, by many orders of magnitude, the probability of dissemination of recombinant DNA outside the experimental area. The fundamental elements of containment include the following:

17. Various combinations of physical and/or biological barriers along with a constant use of standard practices were used to establish different levels of containment for organisms within laboratories (BSL1 through BSL4) and large scale uses (BL1-LS through BL4-LS), plants (BL1-P through BL4-P) and animals (BL1-N though BL4-N).  In all cases, four biosafety levels were established, which provide increasing level of protection to personnel, environment and the community. Categories of containment and biosafety levels are considered separately and detailed descriptions are found in the NIH Guidelines 2002, BMBL (Biosafety in Microbiological and Biomedical laboratories) 2007 and Bangladesh Biosafety Guidelines 2006.

18.  It is important to note that the risk groups correlate with but do not equate to biosafety levels. The risk group of an agent should be one factor, to be considered in association with mode of transmission, procedural protocols, experience of staff, and other factors in determining the BSL in which the work will be conducted. A summary of the characteristics of the four BSLs within laboratory is shown in Table 2 below.

20. Standard Microbiological Practices  Strict adherence  Aware of potential hazard  Trained & proficient in techniques  Supervisors responsible for Appropriate Laboratory facilities Personnel & Training  Special practices & precautions Occupational Health Programs

21. Biosafety Level-1 (BSL-1 or ABSL-1)  Well characterized agents  Agents not known to cause disease in healthy human adults  Prophylactic treatment available  Animals in open cage system or open environment  Risk Group 1 Agents E.coli K-12 Transgenic Plants Plasmids Fungi, Mold and Yeast

22. BSL-1/ABSL-1 Practices  Bench-top work allowed  Daily Decontamination  Manual pipetting  Required Hand washing  Red bag waste  Bio cabinet not required (unless creating aerosols)

23. Biosafety Level-2 (BSL-2 or ABSL-2)  Agents associated with human disease but treatment available  Direct contact /Percutaneous exposure/Mucus membrane exposure  Scratch, Puncture, Needle stick, Eyes, Mouth, open cut  Risk Group 2 Agents Human or Primate Cell Herpes Simplex Virus Attenuated Human Immunodeficiency Virus Patient specimens

24. BSL-2/ ABSL-2 Practices  Limited access to lab when work in progress  Daily decontamination  Mechanical pipetting  Lab coat, safety glasses, gloves required  Red bag & sharps containers required  Biohazard sign posted at entrance to lab  Labeling all equipment  Documented training  Baseline serology or pre- vaccination may be required

25. Biosafety Level 3 (BSL-3 or ABSL-3)  Indigenous or exotic agents Aerosol transmission Serious health effects, treatment may/ may not exist  Risk Group 3 Agents Human Immunodeficiency Virus Mycobacterium tuberculosis Coxiella burnetii

26. BSL-3/ ABSL-3 Practices  Public access NOT permitted  Daily decontamination, autoclave required  Required foot activated hand washing sink and controls  Biohazard signs and labels posted  Air flow from low hazard to high hazard  Bench top work not permitted  Documented training and personnel competency certification (for BSL-3 procedures)

27. Biosafety Level-4 (BSL-4 or ABSL-4)  Dangerous/exotic agents  Life threatening disease  Aerosol transmission  Agents of unknown risk of transmission or health affects  No known treatment  Risk Group 3 Agents Lassa Fever Virus Ebola Hemmorrhagic Fever Virus Marburg Virus Herpes B Virus

28. BSL-4/ ABSL-4 Practices  Builds on BSL-3/ ABSL-3 practices  Maximum containment facilities  Pressurized Containment Suite  BSL-3 + Class III Biosafety Cabinet  Chemical decontamination showers  Liquid effluent collection / decontamination

29. Components of Risk Analysis  Risk analysis is used in its broadest sense as an integrated process consisting of three major components: 1. risk assessment, 2. risk management and 3. risk communication. The individual components are distinct, but are linked to achieve a well-functioning risk analysis process that forms the basis for decision making on any operation or dealing of GMOs. The use of risk analysis in this manner generally conforms to Bangladesh’s proposed NBF and Biosafaty Guidelines, and other national and international principles and guidelines to protect human health and the environment from risks posed by or as a result of modern biotechnology.

30. Risk assessment  Risk assessment is the first and the scientific component of risk analysis. It is a rigorous science-driven process used to identify a hazard and obtain qualitative or quantitative estimate of the levels of risk posed by a hazard including possible adverse effects to human health and the environment.  It typically consists of four steps: (1) hazard analysis (identification and characterization), (2) likelihood estimation, (3) consequence evaluation; and (5) risk estimation.

31. Risk management  Risk management is the second and decision-making component of the process of risk analysis.  It is primarily supported by risk assessment but is informed by other risk considerations.  Risk management is concerned with evaluating whether the risks identified by the risk assessment process are acceptable and manageable, then selecting and implementing the control measures as appropriate to ensure that risks are minimized or controlled

32. Risk communications  Risk communications is recognized as the third component that underpins the risk assessment and risk management processes.  It is the process of exchange of information and opinions concerning risk and risk-related factors among various stakeholders concerned with risk (Codex definition).  It strengthens the over-all process of risk analysis by helping to define the issues and providing the link and the feedback mechanism that informs the two processes.

33.  Risk analysis applied in the broad sense separates the risk assessment from risk management.  The reasons are: to maintain the scientific integrity of the risk assessment, to avoid confusion over the functions to be performed by risk assessors and risk managers, and to minimize any conflict of interest.  In practice, however, this separation is rarely clear-cut and variation in its implementation exists among countries and across regulatory institutions.