Biosafety And Bio Security

BIOSAFETY AND BIOSECURITY IN BILOGICAL LABORATORIES Lt. Gen (R) K.A.Karamat1 Mr. Ejaz Rahim2 Dr. Fazal-e-Hakim Khattak3 Miss. Beenish Mahmood4

INTRODUCTION: Background: Disease diagnosis, human or animal sample analysis, epidemiological studies, scientific research, and pharmaceutical developments: all of these activities are carried out in biological laboratories in the private or public sectors. Biological materials are handled worldwide in laboratories for numerous genuine, justifiable and legitimate purposes. Small and large volumes of living microorganisms are replicated,

where

cellular

components

are

extracted

and

many

other

manipulations undertaken for purposes ranging from educational, scientific, medicinal and health-related to industrial production. Among them, an unknown number of educated people handle, dangerous pathogens or their products every day.

The general public expects laboratory personnel to act responsibly and not to expose the community to bio-risks. They should follow safe working practices (Biosafety) associated with practices that will keep their work and materials safe and secure (Biosecurity), and to follow an ethical code of conduct (bioethics).

However, despite advances in technology, the availability of more and more sophisticated instruments for laboratory use, increasingly effective techniques and the availability of personal protective equipment, human error remains one of the most important factors at the origin of accidents. Poor concentration, denial of responsibilities, incomplete record-keeping, suboptimal facility infrastructure, 1

Advisor (Health) Planning Commission Member (Social Sector) Planning Commission 3 Acting Chief (Health) Planning Commission 4 Intern (Health) Planning Commission 2

1

refusal to acknowledge ethical considerations, lack of codes of conduct, etc. may be at the origin of laboratory-acquired infections.

The data collected from various sources shows that Pakistan has a collection of all kinds of communicable and non communicable diseases. Viral diseases include all types of hepatitis viruses, Encephalitis virus, Polio virus, Congo Crimean Haemorrhagic Fever and Dengue fever viruses while bacterial infections include a range of pathogens from neonatal tetanus to tuberculosis, salmonellae and other pathogens causing acute respiratory infections and diarrhoea. Protozoan and parasitic diseases also contribute a lot to the disease burden. As all kinds of diseases are prevalent in Pakistan with some having pandemic setting and others as epidemic or sporadic, therefore the laboratories both in the public and private sector regularly receive infectious materials as routine specimens and handle them as routine without taking into consideration the bio safety and bio security issues. Disposal of these specimens is also variable with very few labs following the protocols while most just discard it in the sewage.[1]

Laboratory biosafety and Laboratory Biosecurity Laboratory biosafety describes the containment principles, technologies and practices that are implemented to prevent the unintentional exposure to pathogens and toxins, or their accidental release. Laboratory Biosecurity describes the protection, control and accountability for valuable biological materials (VBM) within laboratories, in order to prevent their unauthorized access, loss, theft, misuse, diversion or intentional release. Biosafety aims to prevent the accidental release of pathogens and toxins to laboratory workers, the general population, and the environment. Biosecurity aims to prevent the deliberate theft or diversion of high-risk biological agents for use in bioterrorism or biological weapons proliferation. While the objectives are different, biosafety and biosecurity measures are usually complementary. Biosafety looks at appropriate laboratory procedures and 2

practices necessary to prevent exposures and occupationally acquired infections, while biosecurity addresses procedures and practices to ensure that biological materials and relevant sensitive information remain secure. Purpose of Bio safety and Bio security: As all laboratories, may it be public or private, small or large, rural or urban deal with known or unknown pathogens therefore there is a fear that these pathogens may become a health hazard for the lab workers and for the community if they are not properly handled, stored or discarded. Similarly there is a danger that those working in the labs might get exposed to these known or unknown pathogens without being aware of them or by not taking proper precautions or using personnel protection equipment (PPE).[2] In Pakistan only a few labs are following the biosafety or biosecurity guidelines while majority are not even aware about the basic concept of safety and security. As a starter the purpose of this planning exercise is to formulate a strategic framework for the development of Public sector hospital Laboratories in Pakistan so as to improve/upgrade these laboratories to BSL-2 or more. As a starter one laboratory in each province shall be upgraded to BSL-2 and shall be projected as a model laboratory for others to follow and improve their laboratory set up. All this shall be done with consultation of the provincial heads preferably Director General or Secretary Health.

Pathogens and toxins have been used, even in the recent past, to threaten and harm people, to disrupt society, economies and the political status quo. This has happened in spite of applicable international agreements banning the use of biological agents for malicious use.

3

GOALS:

The goals of the biologicals safety program are to: ¾ Protect staff from exposure to infectious agents ¾ Prevent environmental contamination ¾ Protect experimental materials ¾ Comply with federal and local regulations.

Pakistan Biosafety Rules, 2005 Notified on 21st April, 2005; S.R.O. 336 (I)/2005

The Rules regulate: ► The manufacture, import, and storage of modified organisms and gene technological products for research, whether conducted in laboratories of teaching, research and development institute in public or private sector. ► The import, export, sale and purchase of genetically modified organisms (GMO) for commercial purposes. ► The work involved laboratory work, field trial and commercial release of developed GMOs (plants, animals and micro-organisms). ► The Guidelines establishes the proper procedures to carry out following activities related to GMOs Under safety limits.

¾ Research and development on GMOs and their products ¾ Release of GMOs and products for field trials ¾ Release of GMOs for commercial purposes.[3]

4

Implementation Mechanism

The mechanism of monitoring and implementation of the National Biosafety Guidelines, 2005 is built on the following three tiers as specified in the Biosafety Rules, 2005.

1. National Biosafety Committee (NBC) 2. Technical Advisory Committee (TAC) 3. Institutional Biosafety Committee (IBC) Current situation: The world today is faced with increased risks of exposure to emerging and reemerging pathogens. These originate not only from natural hosts, but from diagnostic

and

research

laboratories

handling

and

manipulating

these

biohazardous organisms. Applied Biosafety and Biosecurity at national and international levels required to ensure the safety of individuals involved with research, and to protect the environment and public health against accidental and intentional release of pathogens and toxins. In recent years, several countries have developed and implemented laboratory bio-security legislation to regulate possession, use and access to biological materials to permit their appropriate use. Despite

the

advances

in

some

advanced

countries,

in

many

under

developing/developing countries and for many laboratories, guidance or specific requirements for the appropriate handling and storage of valuable biological materials (VBM) do not yet exist. Similarly, despite a greater awareness of biosafety and biocontainment practices, handling infectious microorganisms 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. Individuals who work in a

5

laboratory that handles infectious substances are at risk of exposure to the substances they handle. Laboratory-acquired infections are not uncommon.

Situation in Pakistan: In the past century, medical research has led to improved health and increased life expectancy largely because of major stress being given by the public/private sector in preventing and treating infectious diseases. This success has come about through improved hygiene, and increased public awareness. New threats to health continually emerge, however, as bacteria and viruses evolve and are transported to new environments, or develop resistant to drugs and vaccines. Some familiar examples of these so-called emerging or re-emerging infections include

Viral

Hepatitis,

Multi

drug

resistance,

Tuberculosis,

HIV/AIDS,

Hemorrhagic/Dengue fever, and annual outbreaks of gastroenteritis, influenza, typhoid and cholera. Pakistan being a developing country enjoys fewer of the benefits of advanced biological and biomedical research and faces greater risks because of more limited biosafety measures. In recent years, remarkable advances have been made in advanced biological research in the country, which warrants biosafety measures. However, biosafety in Pakistan is at developing stage and biosafety policy and regulations are still evolving

Biohazards and potentially infectious Material A.

Definition

An agent of biological origin that has the capacity to produce deleterious effects on human, i.e. microorganisms, toxins, and allergens derived from those organisms; and allergens and toxins derived from higher plants and animals. B.

Biological Agent Classification-Risk assessment

It is the responsibility of the principal investigator or laboratory director to conduct a risk assessment to determine the proper work practices and containment 6

requirement for work with biohazards material. The risk assessment process should identify features of microorganisms as well as host environmental factors that influence the potential for workers to have a biohazards exposure. This responsibility cannot be shifted to inexperienced or untrained personnel. The principal investigator or laboratory director should consult with a biosafety officer to ensure that the laboratory is in compliance with established guidelines and regulations. When performing a risk assessment, it is advisable to take a conservative approach if there is incomplete information available.

Classification of infective microorganisms by risk groups Infective micro organisms have been classified into different risk groups for laboratory work. Risk Group 1: (No or low community risk). These micro organisms are unlikely to cause human or animal disease. Risk group 2: (Moderate individual risk, low community risk) Pathogen can cause disease but is unlike to have serious hazard to laboratory workers, community, live stock or environment. Laboratory exposure may cause serious infection but effective treatment and preventive measures are available and risk of spread is limited Risk group 3: (High individual risk, low community risk) Pathogen causes serious disease but does not ordinarily spread from one infected person to other. Effective treatment and preventive measures are available Risk group 4: (High individual and community risk) pathogen causes serious disease that can be readily transmitted from one case to other. Effective treatment and preventive measures are not available. Laboratory facilities are designated as basic – Bio safety Level 1 (BSL-1), Bio safety Level 2 (BSL-2), Bio safety Level 3 (BSL-3) and maximum containment Bio safety Level 4 (BSL-4). The Bio safety levels are based on the design of laboratory, construction, containment facilities, equipment, practices and 7

operational procedures required for working with agents of various risk groups (table 1).[4]

Table-1.

Relation of risk groups to biosafety levels, practices and equipment

RISK BIOSAFETY GROUP LEVEL 1 BasicBiosafety Level 1 2 BasicBiosafety Level 2

3

Containment – Biosafety Level 3

4

Maximum containment – Biosafety Level 4

LABORATORY TYPE Basic teaching, Research

LABORATORY practices GMT

SAFETY EQUIPMENT None; open bench work

Primary health Services; diagnostic services, research Special diagnostic services, research Dangerous pathogen units

GMT plus protective clothing, biohazard sign

Open bench plus BSC for potential aerosols

As Level 2 plus special clothing, controlled access, directional airflow As Level 3 plus airlock entry, shower exit, special waste disposal

BSC and/or other primary devices for all activities

Class III BSC, or positive pressure suits in conjunction with Class II BSCs, double ended autoclave (through the wall), Filtered air BSC- biological safety cabinet; GMT= good microbiological techniques.

Countries make their own classification of micro organisms, by risk group, taking into consideration: 1. Pathogenicity of the organism 2. Mode of transmission and host range of organism. These are influenced by immunity levels of the population, density and movement of host population,

presence

of

appropriate

vectors

and

standards

of

environmental hygiene. 3. Local availability of preventive measures like prophylaxis, food and water hygiene, control of vectors 4. Local availability of effective treatment like passive immunization, post exposure vaccination, use of antimicrobials, antivirals and chemotherapy.

8

The bio safety level for a laboratory and its workers must be based on risk assessment and on predefined parameters. The facility requirements at 4 BSLs are shown in table 2. The assignment of BSL takes into consideration the organisms studied, facilities available, equipment practices and procedures required to conduct work in the laboratory. Table - 2.

Summary of biosafety level requirements

BIOSAFETY LEVEL 1 2 3 Isolation of laboratory No No Yes Yes No No Room sealable for decontamination Ventilation: Desirable Yes No - inward airflow Desirable Yes No - controlled ventilating system Desirable Yes/No b No - HEPA-filtered air exhaust Double-door entry No No Yes Airlock No No No Airlock with shower No No No Anteroom No No Yes Anteroom with shower No No Yes/No c Effluent treatment No No Yes/No c Autoclave: - on site No Desirable Yes - in laboratory room No No Desirable - double ended No No Desirable Biological safety cabinets No Desirable Yes Personnel safety monitoring capability d No No Desirable a Environmental and functional isolation from general traffic. b Dependent on location of exhaust. c Dependent on agent(s) used in the laboratory. d For example, window, closed-circuit television, two-way communication.

4 Yes Yes Yes Yes Yes Yes Yes Yes No No Yes Yes Yes Yes Yes

Proper handling and disposal of laboratory materials (specimens) can play a major role in preventing the spread of disease to those who are handling the specimens and also those who accidentally get exposed to these micro organisms while the specimen are being disposed. There is a need to conceptualize a national strategic framework for public sector hospital laboratories in Pakistan for efficient implementation of bio safety and bio security issues. All provinces and other stakeholders have to be involved in

9

various loops of development cycle of this system for generating ownership and mobilizing resources and developing required human resource.

GUIDELINES FOR LABORATORY SAFETY ► All material of human origin is regarded as potentially hazardous. ► Eating, drinking and smoking forbidden in the laboratory. ► All kinds of laboratory record are kept up to dates and in order. ► Warning symbols are pasted out side the entrance of the laboratory.

Clinical Lab safety ¾ Sufficient

safety

equipment

e.g.

firefighting

equipment

(and

its

understanding) safety cabinets, fume cupboards, antidotes & resuscitation apparatus. ¾ Emergency treatment boxes by all departments, locations to be known to all workers, safety officer to be conversant with lab – hazards.

PERSONAL PROTECTION ¾ Laboratory coats should be worn by all while working in the lab. ¾ Hands should be washed after handling chemicals and before leaving the department. ¾ Gloves should be used when handling dangerous materials. ¾ Periodic instructions should be given to the workers regarding good laboratory practice. ¾ Following aspects are important. 1. Sources of laboratory infections 2. Laboratory hazards. 3. Workers rights and duties in relation to safety measures. 4. Access to laboratories 5. Personal hygiene 6. Protective clothing

10

¾ Health education specially in respect of enhancing the awareness of the worker to the danger. ¾ Strict check on entry of unauthorized personnel in the laboratory. ¾ Initial medical examination of all lab personnel/Immunization status. ¾ Periodic medical examination if necessary.

Contingency plans and emergency procedures Every laboratory that works with infective microorganisms should institute safety precautions appropriate to the hazard of the organisms and the animals being handled. A written contingency plan for dealing with laboratory and animal facility accidents is a necessity in any facility that works with or stores Risk Group 3 or 4 microorganisms (containment laboratory – Biosafety Level 3 and maximum containment laboratory – Biosafety Level 4). National and/or local health authorities should be involved in the development of the emergency preparedness plan. Contingency plan The contingency plan should provide operational procedures for: 1. Precautions against natural disasters, e.g. fire, flood, earthquake and explosion 2. Biohazard risk assessment 3. Incident-exposure management and decontamination 4. Emergency evacuation of people and animals from the premises 5. Emergency medical treatment of exposed and injured persons 6. Medical surveillance of exposed persons 7. Clinical management of exposed persons 8. Epidemiological investigation 9. Post-incident continuation of operations. In the development of this plan the following items should be considered for inclusion: 1. Identification of high-risk organisms 2. Location of high-risk areas, e.g. laboratories, storage areas, animal facilities 11

3. Identification of at-risk personnel and populations 4. Identification of responsible personnel and their duties, e.g. biosafety officer, safety personnel, local health authority, clinicians, microbiologists, veterinarians, epidemiologists, and fire and police services 5. Lists of treatment and isolation facilities that can receive exposed or infected persons 6. Transport of exposed or infected persons 7. Lists of sources of immune serum, vaccines, drugs, special equipment and supplies 8. Provision of emergency equipment, e.g. protective clothing, disinfectants, chemical and biological spill kits, decontamination equipment and supplies. Puncture wounds, cuts and abrasions The affected individual should remove protective clothing, wash the hands and any affected area(s), apply an appropriate skin disinfectant, and seek medical attention as necessary. The cause of the wound and the organisms involved should be reported, and appropriate and complete medical records kept. Ingestion of potentially infectious material Protective clothing should be removed and medical attention sought. Identification of the material ingested and circumstances of the incident should be reported, and appropriate and complete medical records kept. Potentially infectious aerosol release (outside a biological safety cabinet) All persons should immediately vacate the affected area and any exposed persons should be referred for medical advice. The laboratory supervisor and the biosafety officer should be informed at once. No one should enter the room for an appropriate amount of time (e.g. 1 h), to allow aerosols to be carried away and heavier particles to settle. If the laboratory does not have a central air exhaust system, entrance should be delayed (e.g. for 24 h).

12

Signs should be posted indicating that entry is forbidden. After the appropriate time, decontamination should proceed, supervised by the biosafety officer. Appropriate protective clothing and respiratory protection should be worn. Broken containers and spilled infectious substances Broken containers contaminated with infectious substances and spilled infectious substances should be covered with a cloth or paper towels. Disinfectant should then be poured over these and left for the appropriate amount of time. The cloth or paper towels and the broken material can then be cleared away; glass fragments should be handled with forceps. The contaminated area should then be swabbed with disinfectant. If dustpans are used to clear away the broken material, they should be autoclaved or placed in an effective disinfectant. Clothes, paper towels and swabs used for cleaning up should be placed in a contaminated-waste container. Gloves should be worn for all these procedures. If laboratory forms or other printed or written matter are contaminated, the information should be copied onto another form and the original discarded into the contaminated-waste container.

Breakage of tubes containing potentially infectious material in centrifuges not having sealable buckets

If a breakage occurs or is suspected while the machine is running, the motor should be switched off and the machine left closed (e.g. for 30 min) to allow settling. If a breakage is discovered after the machine has stopped, the lid should be replaced immediately and left closed (e.g. for 30 min). In both instances, the biosafety officer should be informed. Strong (e.g. thick rubber) gloves, covered if necessary with suitable disposable gloves, should be worn for all subsequent operations. Forceps, or cotton held in the forceps, should be used to retrieve glass debris. All broken tubes, glass fragments, buckets, and the rotor should be placed in a non corrosive disinfectant known to be active against the organisms concerned

13

Unbroken, capped tubes may be placed in disinfectant in a separate container and recovered. The centrifuge bowl should be swabbed with the same disinfectant, at the appropriate dilution, and then swabbed again, washed with water and dried. All materials used in the clean-up should be treated as infectious waste. Breakage of tubes inside sealable buckets (safety cups) All sealed centrifuge buckets should be loaded and unloaded in a biological safety cabinet. If breakage is suspected within the safety cup, the safety cap should be loosened and the bucket autoclaved. Alternatively, the safety cup may be chemically disinfected. Fire and natural disasters Fire and other services should be involved in the development of emergency preparedness plans. They should be told in advance which rooms contain potentially infectious materials. It is beneficial to arrange for these services to visit the laboratory to become acquainted with its layout and contents. After a natural disaster, local or national emergency services should be warned of the potential hazards within and/or near laboratory buildings. They should enter only when accompanied by a trained laboratory worker. Infectious materials should be collected in leak proof boxes or strong disposable bags. Salvage or final disposal should be determined by biosafety staff on the basis of local ordinances. Emergency services: whom to contact The telephone numbers and addresses of the following should be prominently displayed in the facility: 1. The institution or laboratory itself (the address and location may not be known in detail by the caller or the services called) 2. Director of the institution or laboratory/ Medical officer 3. Laboratory supervisor 4. Biosafety officer 5. Fire services 14

6. Hospitals/ambulance services/medical staff (names of individual clinics, departments, and/or medical staff, if possible) 7. Police 8. Water, gas and electricity services.

REFERENCES 1. PC-1 of Establishment of Biosafety/ Biosecurity system for public sector Hospital/ Health institution dealing with infectious materials in Pakistan-2005 2. Strategic Frame work for Biosafety and Biosecurity in Public sector Hospitals I Pakistan, PMRC-Ministry of Health. 3. National Biosafety guidelines, Ministry of Enviornment-2005 4. Biological safety Manual, Third Edition-2005. 5. Laboratory Biosafety manual, World Health Organization-2004 6. Biosafety in Microbiological and Biomedical Laboratories 4th Edition: http://www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm 7. http://cns.miis.edu/research/cbw/biosec/pdfs/biosec.pdf 8. http://www.fas.org/programs/ssp/bio/resource/biosafetylevels.html

15