B IO- S AFETY G UIDELINES

Contents:

1. Introduction2. History3. Background4. Purpose5. Basic Principles6. Biological Waste & Its Disposal7. Risk Groups 8. Bio-Safety Levels9. Risk Assessment10.Conclusion

Abstract

These guidelines for bio-safety laboratory competency outline the essential skills, knowledge, and abilities required for working with biologic agents at the four bio-safety levels (BSLs) (levels 1, 2, 3, and 4). This article comprises all the basic guidelines necessary to carry out any type of research being carried out on biological organisms carefully and without harming anyone i.e. the researcher, workers and co-workers and most importantly our environment and also it carries guidelines to handle bio-hazardous materials and dispose them off according to the protocol.

For a person with no scientific background bio-hazardous symbols have also been discussed so that when they see those symbols they would be able to protect themselves accordingly.

In the end a conclusion has been given which consists the essence of the discussion of the whole research paper.

1. Introduction:

Bio-safety is the prevention of large scale loss of biological integrity, focusing both on ecology and human health. These prevention mechanisms include conduction of regular reviews of the bio-safety in laboratory settings, as well as strict guidelines to follow. Bio-safety is used to protect us from harmful incidents. High security facilities are necessary when working with Synthetic Biology as there are possibilities of bioterrorism acts or release of harmful chemicals and or organisms into the environment. A complete understanding of experimental risks associated with synthetic biology is helping to enforce the knowledge and effectiveness of bio-safety. 1

Despite a greater awareness of bio-safety and bio-containment practices, handling infectious microorganisms and other organisms remains a source of infection, and even mortality, among laboratory workers. Incidents of secondary transmission of disease to the public at large, which may be due to possible contamination of the environment or personnel, are also occurring. Laboratory workers can minimize the risks associated with work involving these infectious agents through the application of appropriate bio-safety and containment principles and practices.

Bio-safety laboratories must ensure adequate safety conditions to avoid potential hazards associated with the handling of biologic materials, the manipulation of genomes, the creation of synthetic organisms, and the spread of multidrug-resistant bacteria, and threats of biologic terrorism. These guidelines define the essential competencies needed by laboratory personnel to

work safely with biologic materials and other hazards that might be found in a biologic laboratory (e.g. those related to research animals, chemicals, radiologic materials, and the physical environment).

A successful laboratory safety program encompasses a continuous process of hazard recognition, risk assessment, and hazard mitigation. The risk for exposures, laboratory acquired infections, and the unintended release of research or clinical materials to the environment should ultimately be reduced by ensuring the competency of laboratories at all levels.

Based on this above discussion bio-safety can be defined as “procedures or measures designed to protect the population against harmful biological or biochemical substances”.2

2. History:

On 18 April 1955 the first biological safety conference took place at Camp Detrick and involved members of the military representing Camp Detrick, Pine Bluff Arsenal, Arkansas (PBA), and Dug way Proving Grounds, Utah (DPG). In that conference role of Safety in the Biological Warfare, chemical, radiological and industrial safety issues were discussed. Later in the US, the Centre for Disease Control (CDC) specified 4 different levels of bio-containment which ranges from bio-safety level 1 to bio-safety level 4. 3 4

3. Background:

The Laboratory bio-safety guidelines were initially developed to guide government, industry, university, hospital, and other public health and microbiological laboratories in their development

of bio-safety policies and programs. The Guidelines also serve as a technical document providing information and recommendations on the design, construction and commissioning of containment facilities.5 Several other circumstances led to the development of bio-safety competencies for practicing laboratorians. In 2006, PAHPA called for the assessment, development, delivery, and evaluation of competency-based training for bio-safety in high containment laboratories. In 2008, CDC convened the Blue Ribbon Panel for issues of Clinical Laboratory Safety to address incidence of laboratory-acquired infection. Also in 2008, a Trans-Federal Task Force was convened for federal agencies with laboratories to address bio-safety and bio-security in working with agents that pose a significant public health threat, whether they arise from nature, accidental exposure, or deliberate terrorist attacks. These efforts underscored the need to develop competency guidelines for laboratory bio-safety.

4. Purpose:

The primary purpose of these guidelines is to establish the behaviors and knowledge that laboratory workers at all levels should have to work safely with biologic materials. Other key issues are: the need for a well-designed workspace, knowledge of specific biologic agents and toxins, quality laboratory management practices, and an overall safety culture.

5. Basic Principles:

No regulation or guideline can ensure safe practices. Individual and organizational attitudes regarding safety will influence all aspects of safe practice, including willingness to report

concerns, response to incidents, and communication of risk. Each organization should strive to develop a culture of safety that is open and nonpunitive, encourages questions, and is willing to be self-critical. Persons and organizations must be committed to safety, be aware of risks, behave in ways that enhance safety, and be adaptable. Scientists understand that practices should be refined as observations are made, hypotheses tested, findings published, and technical progress achieved. The same holds true for safety in the laboratory, which should evolve as experience is gained and as laboratory activities change. As laboratorians gain more knowledge over time concerning how to recognize and control hazards, the level of risk that is considered acceptable should become smaller, with the goal of moving continuously to eliminate or reduce risk to the lowest reasonably achievable level.

Laboratorians have both the responsibility to report concerns to management and the right to express concerns without fear of reprisal. Similarly, management has the responsibility to address concerns raised from any direction. A continuous process of hazard recognition, risk assessment, and hazard mitigation practices ensures that management and laboratory workers alike are aware of the issues and work together to maintain the highest standard of safety.

6. Bio-Hazardous Waste & Its Disposal:

Bio-hazardous waste:- Bio-hazardous waste is defined as materials

containing:

Infectious agents (to human, plants, animals)

Biological toxins

Materials derived from humans and primates (blood, body fluids, tissues)

Human and primate cell lines (including recombinant)

Recombinant animal cell lines

Recombinant microorganisms

Transgenic animals (vertebrate and invertebrate)

Materials derived from transgenic animals (body fluids, tissues)

Transgenic plants

Recombinant materials such as plasmids, DNA/RNA, synthetic DNA

Disposal Of Bio-Hazardous Waste:-

Liquid Waste:Bio-hazardous liquid wastes are liquids

containing bio-hazardous waste.

Disposal Method: Decontaminate by treating with an appropriate disinfectant. The amount of contact time will depend on the chemical used and the material decontaminated. For specific procedures, refer to your approved protocol or contact EHS.

Solid Waste:Bio-hazardous solid wastes are solids that

contain bio-hazardous materials or lab waste that has come in contact with bio-hazardous materials. These include:

Culture media

Personal Protective Equipment (contaminated)

Plastic ware including pipette tips

Transgenic plants including soil

Disposal Methods: Place material in an autoclave bag, close bag loosely; attach a strip of autoclave tape and autoclave. Apply “treated” label and place in black garbage bag for disposal in trash. Record treatment on autoclave waste treatment record form.

Autoclave bag

Autoclave bag with “treated” label attached.

Treated bag placed in black trash bag for disposal.

Or; Place in biohazard waste container and contact EHS for pickup.

Sharps: Dispose into sharps container

Glassware: Decontaminate for re-use or dispose in sharps container. Do not use glass trash boxes for glassware contaminated with biological materials.

Transgenic Animals and Materials: Transgenic animals and materials

includes animals (vertebrate and invertebrate) and materials (tissues and infected bedding) from transgenic animals.

Transgenic insects:

Freeze, then autoclave or place in biohazard container

Transgenic animal carcasses/tissues:

Place in 2X biohazard bags and place in freezer

Animal bedding from animals shedding pathogens:

Place in biohazard waste container.

Disposal Method: Place in appropriate container. Request EHS pickup.

Non-transgenic Animals/Tissues: Non-transgenic animals or tissues

include animal carcasses/tissues not infected with infectious agents.

Disposal Method: Place in double bags and place material in freezer and request EHS pickup.

Used Animal Bedding: Used animal bedding includes bedding

from animals that is free of pathogens or biological toxins

Disposal Method: Can be disposed as conventional trash. Containers should be secured.6

7. Risk Groups:

Classification of organisms according to risk group has traditionally been used to categorize the relative hazards of infective organisms. The factors used to determine which risk group an organism falls into is based upon the particular characteristics of the organism, such as

pathogenicity

infectious dose

mode of transmission

host range

availability of effective preventive measures

availability of effective treatment

These classifications presume ordinary circumstances in the research laboratory or growth in small volumes for diagnostic and experimental purposes. Four levels of risk have been defined as follows.

Risk Group 1 (low individual and community risk):

Any biological agent that is unlikely to cause disease in healthy workers or animals.

Risk Group 2 (moderate individual risk, low community risk):

Any pathogen that can cause human disease but, under normal circumstances, is unlikely to be a serious hazard to laboratory workers, the community, livestock or the environment. Laboratory exposures rarely cause infection leading to serious disease; effective treatment and preventive measures are available, and the risk of spread is limited.

Risk Group 3 (high individual risk, low community risk):

Any pathogen that usually causes serious human disease or can result in serious economic consequences but does not ordinarily spread by casual contact from one individual to another, or that causes diseases treatable by antimicrobial or anti-parasitic agents.

Risk Group 4 (high individual risk, high community risk):

Any pathogen that usually produces very serious human disease, often untreatable, and may be readily transmitted from one individual to another, or from animal to human or vice-versa, directly or indirectly, or by casual contact.7

8. Bio-Safety Levels: There are 4 types of bio-safety levels

according to the risk factors involved depending on the nature of pathogen being handled.

Bio-Safety Level 1 (BSL 1): This applies to the basic

laboratory that handles agents requiring bio-safety level 1. BSL 1 requires no special design features beyond those suitable for a well-designed and functional laboratory. Biological safety cabinets (BSCs) are not required. Work may be done on an open bench top, and containment is achieved through the use of practices normally employed in a basic microbiology laboratory.

Bio-Safety Level 2 (BSL 2):

This applies to the laboratory that handles agents requiring bio-safety level 2. The primary exposure hazards associated with organisms requiring BSL 2 are through the ingestion, inoculation and mucous membrane route. Agents requiring BSL 2 facilities are not generally transmitted by airborne routes, but care must be taken to avoid the generation of aerosols (aerosols can settle on bench tops and become an ingestion hazard through contamination of the hands) or splashes. Primary containment devices such as BSCs and centrifuges with sealed rotors or safety cups are to be used as well as appropriate personal protective equipment (i.e., gloves, laboratory coats, protective eyewear). As well, environmental contamination must be minimized by the use of hand washing sinks and decontamination facilities (autoclaves).

Bio-Safety Level 3 (BSL 3):

This applies to the laboratory that handles agents requiring bio-safety level 3. These agents may be transmitted by the airborne route, often have a low infectious dose to

produce effects and can cause serious or life-threatening disease. BSL3 emphasizes additional primary and secondary barriers to minimize the release of infectious organisms into the immediate laboratory and the environment. Additional features to prevent transmission of BSL3 organisms are appropriate respiratory protection, HEPA filtration of exhausted laboratory air and strictly controlled laboratory access.

Bio-Safety Level 4 (BSL 4):

This is the maximum containment available and is suitable for facilities manipulating agents requiring bio-safety level 4. These agents have the potential for aerosol transmission, often have a low infectious dose and produce very serious and often fatal disease; there is generally no treatment or vaccine available. This level of containment represents an isolated unit, functionally and, when necessary, structurally independent of other areas. BSL 4 emphasizes maximum containment of the infectious agent by complete sealing of the facility perimeter with confirmation by pressure decay testing; isolation of the researcher from the pathogen by his or her containment in a positive pressure suit or containment of the pathogen in a Class III BSC line; and decontamination of air and other effluents produced in the facility.

9. Risk Assessment:

Risk assessment is a critical step in the selection of an appropriate bio-safety level for the microbiological work to be carried out. A detailed local risk assessment should be conducted to determine whether work requires containment level 1, 2, 3 or 4 facilities and operational practices. Individuals with varying

expertise and responsibilities should be included in the risk assessment process and can include, among others, the facility director, laboratory supervisor, principal investigator, senior microbiologist, bio-safety officer and bio-safety committee.

Available information can be used as a starting point to assist in the identification of risk factors, including the recommended Risk Group of the organism. In addition to the Risk Group classifications, which are based on the risk factors inherent to the organism, the following factors associated with the laboratory operation should also be examined:

potential for aerosol generation

quantity

concentration

agent stability in the environment (inherent biological decay rate)

type of work proposed (e.g., in vitro , in vivo , aerosol challenge studies)

use of recombinant organisms (e.g., gene coding for virulence factors or toxins; host range alteration; oncogenicity; replication capacity; capability to revert to wild type)

10. Conclusion: These guidelines outline the essential

expectations for behaviors and knowledge of laboratory workers necessary to work safely with biologic materials at all levels of the profession in the life sciences. The development of these guidelines is a first step toward defining comprehensive safety competencies in biologic laboratories. These guidelines reflect a range of past experiences and will be reviewed periodically and refined as additional experience is gained. The guidelines can be used as a resource to develop

educational goals, training standards, safety assessments, professional development, and certification.

Every organization using these competencies should regularly review and improve its practices and documents with an eye toward continual reduction of the risks involved in working with biologic and other hazardous laboratory materials. Training is not limited to the initial instruction received at the start of a laboratory worker’s employment but is continuous and refreshed periodically. Some professions or organizations that were contributors to these guidelines, including AALAS and the Council of State and Territorial Epidemiologists, also have addressed biologic safety practices. 8

1 http://en.wikipedia.org/wiki/Biosafety 2 https://www.google.com.pk/?gws_rd=cr&ei=y2PcU5rOG6mh0QX4s4CQBg#q=define+biosecurity 3 http://www.slideshare.net/neharachankar/neha-biosafety-levels-ppt 4 http://fas.org/pubs/pir/2011fall/2011fall-bioagents.pdf 5 http://www.phac-aspc.gc.ca/publicat/lbg-ldmbl-04/ch1-eng.php 6 http://www.utexas.edu/safety/ehs/biosafety/waste_guidelines.html 7 http://www.phac-aspc.gc.ca/publicat/lbg-ldmbl-04/ch2-eng.php 8 https://www.aaalac.org/accreditation/RefResources/guide_for_biosaf_comp.pdf