LESSON 6

Basic Concepts on laboratory Biosafety and Biosecurity

Brief History of Laboratory Biosafety

Observing and implementing laboratory safety precautions are of utmost importance in the
medical technology practice. Individuals who handle and process microbiological specimen are
vulnerable to pathogenic microorganisms which are possible sources of laboratory acquired
infections (LAI).

Laboratory biosafety and biosecurity traces its history in North America and Western Europe. The
origins of biosafety is rooted in the US biological weapons program which began in 1943, as
ordered by then US President Franklin Roosevelt and was active during the Cold War. It was
eventually terminated by US President Richard Nixon in 1969. In 1943, Ira L Baldwin became the
first scientific director of Camp Detrick (which eventually became Fort Detrick), and was tasked
with establishing the biological weapons program for defensive purposes to enable the United
States to respond if attacked by such weapons. After the Second World War. Camp Detrick was
designated a permanent installation for biological research and development Biosafety was an
inherent component of biological weapons development. Later on. Newell A. Johnson designed
modifications for biosafety at Camp Derrick, He engaged some of Camp Detrick’s leading
scientists about the nature of their work, and developed specific technical solutions such as Class
III safety cabinets and laminar flow hoods to address specific risks. Consequent meetings
eventually led to the formation of the American Biological Safety Association (ARS) in 1989. The
association held annual meetings that soon became the ABSA annual conferences (Salerno et al.,
2015)

Other contributors outside the United States included Arnold Wedum who described the use of
mechanical pipettors to prevent laboratory-acquired infections in 1907 and 1908 (Kruse (1991),
cited by Salerno, 2015). Moreover, ventilated cabinets, early progenitors to the nearly ubiquitous
engineered control now known as the biological safety cabinet, were also first documented
outside of the US biological weapons program. In 1909, a pharmaceutical company in
Pennsylvania developed a ventilated cabinet to prevent infection from mycobacterium
tuberculosis.

At the height of increasing mortality and morbidity due to smallpox in 1967, WHO aggressively
pursued the eradication of the virus (College of Physicians of Philadelphia 2014). It was also
during this time that serious concerns about biosafety practices worldwide were raised,
contributing directly to the decision of the World Health Assembly to consolidate the remaining
virus stocks into two locations: the Center for Disease Control and Prevention (CDC) in the United
States and the State Research Center of Virology and Biotechnology VECTOR (SRCVB VECTOR) in
Russia. In 1974. The CDC published the Classification of Etiological Agents on the Basis of Hazard
that introduced the concept of establishing ascending levels of containment associated with risks
in handling groups of infectious microorganisms that present similar characteristics. Two years
later, the National Institutes of Health (NIH) of the United States published the NIH Guidelines for
Research Involving Recombinant DNA Molecules. It explained in detail the microbiological
practices, equipment, and facility necessarily corresponding to four ascending levels of physical
containment.

The guidelines laid the foundation for the introduction of a code of biosafety practice. The code,
along with WHO first edition of Laboratory Biosafety Manual (1983) and the CDC and NIH’s
jointly-published first edition of the Biosafety in Microbiological and Biomedical Laboratories
(1984), marked the development of the practice of laboratory biosafety. These documents
established the model of biosafety containment levels with certain agents which increased the
biosafety levels for biological agents that pose risks to human health. Biosafety levels are the
technical means of mitigating the risk of accidental infection from or release of agents in the
laboratory setting as well as the community and environment it is situated in. Although biosafety
levels are concentrated in a combination of engineered control, administrative controls, and
practice, the emphasis is clearly on the equipment and facility control with little attention given
to risk assessment.
This progress in biosafety practice continued until the emergence of a community of “biosafety
officers” who adopted the administrative role of ensuring that the proper equipment and facility
controls are in place based on the specified biosafety level of the laboratory.

Arnold Wedum, director of Industrial Health and Safety at the US Army Biological Research
Laboratories in 1944, was recognized as one of the pioneers of biosafety the provided the
foundation for evaluating the risks of handling infectious microorganisms and for recognizing
biological hazards and developing practices, equipment, and facility safeguards for their control.
In 1966, Wedum and microbiologist Morton Reitman, colleagues at Fort Detrick, analyzed
multiple epidemiological studies of laboratory-based outbreaks

Brief History of Laboratory Biosecurity

In 1996, the US government enacted the Select Agent Regulations to monitor the transfer of a
select list of biological agents from one facility to another. Slightly after the terrorist attacks and
the anthrax attacks of 2001. Also known as Amerithrax, the US government changed its
perspective. The revised Select Agent Regulations then required specific security measures for
any facility in the United States that used or stored one or more agents on the new, longer list of
agents.

The revision of the Select Agent Regulations in 2012 sought to address the creation of two tiers
of select agents, Tier 1 agents are materials that pose the greatest risk of deliberate misuse, and
the remaining select agents. This change was intended to make the regulations more risk-based,
mandating additional security measures for Tier 1 agents. Other countries also relatively
implemented and prescribed biosecurity regulations for bioscience facilities Singapore’s
Biological Agents and Torinx Act is similar in scope with the US regulations but with more severe
penalties for noncompliance (Republic of Singapore 2005). In South Korea the Act on Prevention
of Infectious Diseases in 2005 was amended to require institutions that work with listed “highly
dangerous pathogens” to implement laboratory biosafety and biosecurity requirements to
prevent the loss, theft, diversion, release, or misuse of these agents, In Japan, the Infectious
Disease Control Law was recently amended under Japans Ministry of Health, Labor, and Welfare.
It also established four schedules of select agents that are subject to different reporting and
handling requirements for possession, transport. And other activities. Then in Canada, Canadian
containment level (CL) 3 and CIA facilities that work with risk group 3 or 4 are required to undergo
certification. In 2008, the Danish Parliament passed a law that gives the Minister of Health and
Prevention the authority to regulate the possession, manufacture, use, storage, sale, purchase
or other transfer, distribution, transport, and disposal of listed biological agents. Around the
world, biosecurity implementation has become a purely administrative activity based on a
government developed checklist.

Local and International Guidelines on Laboratory Biosafety and Biosecurity

In February 2008, the Comité Européen de Normalisation (CEN), a European Committee for
Standardization published the CEN Workshop Agreement 15793 (CWA 15793) which focuses on
laboratory biorisk management. The Workshop offers a mechanism where stakeholders can
develop consensus standards and requirements in an open process. The CWA 15793 can be
applied to international stakeholders, however, they do not have the force of regulation while
conformity is voluntary. The CWA 15793 was developed among experts from 24 different
countries including Argentina, Australia, Belgium, Canada, China, Denmark, Germany Ghana, UK,
US, among others. It was updated in 2011 and intended to maintain a biorisk management
system among diverse organizations and set out performance-based requirements with the
exclusion of guidance for implementing a national biosafety system. Since it originated in the
European workshop agreement framework, confusion among countries outside Europe arose
especially in the United States in terms of its applicability Nevertheless, the agreement was used
until it officially expired in 2014 (Gronvall. 2015).

To address concerns on biosafety guidance for research and health laboratories, issues on risk
assessment and guidance to commission and certify laboratories, the WHO in 1983 published its
3rd edition of the Laboratory Biosafety Manual. It includes information on the different levels of
containment laboratories (Biosafety levels 1-4), different types of biological safety cabinets, good
microbiological techniques, and how to disinfect and sterilize equipment. In terms of biosecurity,
it covers the packaging required by international transport regulations and other types of safety
procedures for chemical, electrical, ionizing radiation, and fire hazards. The manual puts
emphasis on the continuous monitoring and improvement directed by a biosafety officer and the
biosafety committee. Unfortunately, there is no mechanism to ensure that the WHO biosafety
guidance is being adhered to, or that people working in laboratories are sufficiently trained.

The Cartagena Protocol on Biosafety (CPB), made effective in 2003 which applies to the 165
member-countries provides an international regulatory framework to ensure an adequate level
of protection in the field of safe transfer, handling, and use of living modified organisms (LMOs)
resulting from modern biotechnology.” The regulations primarily tackle the safe transfer,
handling, and use of L.MOs that may have adverse effects on the conservation of biological
diversity except those that are used for pharmaceuticals purposes. In addition, the legislation
provides a framework for assessing the risk of LOSS and is focused on ensuring that LMOs do not
negatively affect biodiversity.

The new National Committee on Biosafety of the Philippines (NCBP) established under E.O. 430
series of 1990 was formed on the advocacy efforts of scientists. The mandate of NCPB focuses
on the organizational structure for biosafety: procedures for evaluation of proposals with
biosafety concerns: procedures and guidelines on the introduction, movement, and field release
of regulated materials: and procedures on physico-chemical and biological containment. On
March 17, 2006, the Office of the President promulgated E.O. 514 establishing the National
Biosafety Framework (NBF), which prescribes the guidelines for its implementation,
strengthening the National Committee on Biosafety of the Philippines, The NBF is a combination
of policy, legal, administrative, and technical instruments developed to attain the objective of
the Cartagena Protocol on Biosafety which the Philippines signed on May 24, 2000. The NBF can
be considered as an expansion of the NCBP, which since 1987 has played an important role in
pioneering the establishment and development of the current biosafety system of the country
and was acknowledged as a model system for developing countries. The Department of
Agriculture (DA) also issued Administrative Order No. 8 to set in place policies on the importation
and release of plants and plant products derived from modern biotechnology. The Department
of Health (DOH), together with NCBP, formulated guidelines in the assessment of the impacts on
health posed by modern biotechnology and its applications. The guidelines aid in evaluating and
monitoring processed food derived from or containing GMO. Currently, DOH, in the midst of
technological advances, recognizes the need to update the minimum standards and technical
requirements for clinical laboratories, It requires clinical laboratories to ensure policy guidelines
on laboratory biosafety and biosecurity (DOH Administrative Order No. 2007-0027).

Different Organizations in the Field of Biosafety

Several organizations across continents have undertaken initiatives in advocating for laboratory
biosafety and biosecurity. The following are some prominent organizations inside and outside
the Philippines:

1. American Biological Safety Association (ABSA) a regional professional society for

biosafety and biosecurity founded in 1984. It promotes biosafety as a scientific discipline
and provides guidance to its members on the regulatory regime present in North America.
2. Asia-Pacific Biosafety Association (A-PBA) a group founded in 2005 that acts as a
professional society for biosafety professionals in the Asia-Pacific region. Its members are
from Singapore, Brunei. China, Indonesia, Malaysia, Thailand, the Philippines, and
Myanmar. Active members of the International Biosafety Working Group are required to
directly contribute to the development of the best biosafety practices.
3. European Biological Safety Association (EBSA) a non-profit organization founded in June
1996, that aims to provide a forum for discussions and debates on issues of concern and
to represent those working in the field of biosafety. EBSA focuses on encouraging and
communicating among its member’s information and issues on biosafety and biosecurity
as well as emerging legislation and standards.
4. Philippine Biosafety and Biosecurity Association ( PBA) rated by a multi disciplinary team
with members coming from the health and education sectors as well as individuals from
the executive, legislative, and judicial branches of the government. Also included are
members of the steering committee and technical working groups of the National
Laboratory Biosafety and Biosecurity Action Plan Task Force established as per DPO No.
 2006-2500 dated September 15, 2006. A long term goal of the association is to assist the
DA and DOH in their efforts to create a national policy and implement plan for laboratory
biosafety and biosecurity.
5. Biological Risk Association Philippines (BRAP) a non-government and non-profit
association that works to serve the emergent concerns of biological risk management in
various professional fields such as in the health, agriculture, and technology sectors
throughout the country. It has launched numerous activities in cooperation and
collaboration with other associations, on a national and international scale in the
promotion of biosafety, biosecurity and biorisk management as scientific disciplines.
BRAP goes by the tagline. “assess, mitigate monitor.”

Currently, member countries of ABSA, A-PBA, and EBSA have founded organizations in their
respective nations which share the same goals and objectives in addressing issues and concerns
related to biosafety and biosecurity.

Fundamental Concepts of Laboratory Biosafety and Biosecurity

WHO issued a common understanding of biosafety derived from the practical guidance I
techniques to be used in laboratories. Biosafety has long been practiced in most nations
especially among institutions that handle and process microbiological specimen. The WHO
Laboratory Biosafety Manual (LBM) defines biosafety is “the containment principles,
technologies, and practices that are implemented to prevent unintentional exposure to
pathogens and toxins, or their accidental release." On the other hand, biosecurity refers to the
protection, control, and accountability for valuable biological materials within laboratories, in
order to prevent their unauthorized access, loss, theft, misuse, diversion, or intentional release"
(WHO, 2006). By simple definition, "biosafety protects people from germs" while "biosecurity
protects germs from people."
In 1966, Charles Baldwin, an environmental health engineer working for the Dow Chemical
Company containment systems products, created the biohazard symbol used in labeling
biological materials carrying significant health risks.

Biosafety and biosecurity share common perspectives in terms of risk assessment and
management methodologies, personnel expertise and responsibility, control and accountability
for research materials including microorganisms and culture stocks, access control elements,
material transfer documentation, training, emergency planning, and program management
among others.

To sum up, biosafety focuses on laboratory procedures and practices necessary to prevent
exposure to and acquisition of infections while the maintenance of secure procedures and
practices in handling biological materials and sensitive information falls under biosecurity.

Classifications of Microorganisms According to Risk Groups

WHO recommends an agent risk group classification for laboratory use that describes four
general risk groups based on principal characteristics and relative hazards posed by infectious
toxins or agents. Risk group classification for humans and animals is based on the agent's
pathogenicity, mode of transmission, host range, and the availability of preventative measures
and effective treatment. Through the classification, infective microorganisms are classified as
Risk Group 1, Risk Group 2, Risk Group 3, and Risk Group 4:

1. Risk group 1 - includes microorganisms that are unlikely to cause human or animal
disease. These microorganisms bring about low individual and community risk.
2. Risk group 2 - includes microorganisms that are unlikely to be a significant risk to
laboratory workers and the community, livestock, or the environment. Laboratory
exposure may cause infection, however effective treatment and preventive measures are
available while the risk of spread is limited. This risk group bring about moderate
individual risk and limited community risk.
 3. Risk group 3 - includes microorganisms that are known to cause serious diseases to
humans or animals and may present a significant risk to laboratory workers. It could
present a limited to moderate risk if these microorganisms spread in the community or
the environment, but there are usually effective preventive measures or treatment
available. They bring about high individual risk, and limited to moderate community risk.
4. Risk group 4 - includes microorganisms that are known to produce life-threatening
diseases to humans or animals. It represents a significant risk to laboratory workers and
may be readily transmissible from one individual to the while effective treatment and
preventive measures are not usually available. In effect, they bring about high individual
and community risk.

Categories of Laboratory Biosafety According to Levels

In order to facilitate precautionary measures, CDC categorized laboratories into our biosafety
levels-Biosafety Level 1, Biosafety Level 2, Biosafety Level 3, and Biosafety Level 4. Biosafety level
designations are based on a composite of the design features, construction, continent facilities,
equipment, practices, and operational procedures required for working with from the various
risk groups. They are designated in ascending order, by degree of protection provided to the
personnel the environment, and the community (BMBL, 5th edition).

1. Biosafety Level 1 (BSL-1) is suitable for work involving viable microorganisms that are
defined and with well-characterized strains known not to cause disease in humans.
Examples of microorganisms being handled in this level are Bacillus subtilis, Naegleria
gruberi, infection canine hepatitis virus, and exempt organisms under the NIH Guidelines.
This level is the most appropriate among undergraduate and secondary educational
training and teaching laboratories that require basic laboratory safety practice, safety
equipment, and facility design that requires basic level of containment.
2. Biosafety Level 2 (BSL-2) basically designed for laboratories that deal with indigenous
moderate-risk agents present in the community. It observes practice, equipment, and
facility design that are applicable to clinical, diagnostic, and teaching laboratories
 consequently observing good microbiological techniques Examples of microorganisms
that could be handled under this level are Hepatitis B virus HIV, salmonellae, and
Toxoplasma species. BSL-2 is appropriate when work in done with human blood, body
fluids, tissues, or primary human cell lines where there is uncertain presence of infectious
agents. Hand washing sinks and waste decontamination facilities must be available and
access to the laboratory must be restricted when work is being conducted. All procedures
where infectious aerosols or splashes may be created are conducted in biosafety cabinets
or other physical containment equipment.
3. Biosafety Level 3 (BSL-3) puts emphasis on primary and secondary barriers in the
protection of the personnel, the community, and the environment from Infections
aerosol exposure. Work with indigenous or exotic agents with a potential for respiratory
transmission, and that may cause serious and potentially lethal infection are being
conducted here. Examples of microorganisms handled here are Mycobacterium
tuberculosis, St. Louis encephalitis virus, and Coxiella. All laboratory activities are required
to be performed in a biosafety cabinet or other containment equipment like you tight
aerosol generation chamber. Secondary barriers for this level are highly required
including controlled access to the laboratory and ventilation requirements to minimize
the release of infectious aerosols from the laboratory while special engineering and
design features are being considered. Personnel must be supervised by scientists
competent in handling infectious agents and associated procedures in a BSL-3 laboratory.
4. Biosafety Level 4 (BSL-4) is required for work with dangerous and exotic agents that pose
high individual risks of life-threatening diseases that may be transmitted via the aerosol
route, for which there are no available vaccines or treatment. Specific practices, safety
equipment, and appropriate facility design and construction are required for instance
when manipulating viruses such as the Marburg or the Crimean-Congo hemorrhagic fever
and any other agents known to pose a high risk of exposure and infection to laboratory
personnel, community, and environment. The laboratory worker's complete isolation
from aerosolized infectious materials is accomplished primarily by working in a Class III
biosafety cabinet or in a full body, air-supplied positive-pressure personnel suit. A BSL-4
laboratory is generally a separate building or completely isolated zone with specialized
ventilation requirements and waste management systems. Laboratory staff must have
specific and thorough training in handling extremely hazardous infectious agents. The
laboratory is controlled by the laboratory supervisor in accordance with institutional
policies.