The Evolving Landscape of Biosafety and Biosecurity: A Review of International Guidelines and Best Practices

safeguarding, regulating, and responsibly managing crucial biological materials within a laboratory to prevent unauthorized access, loss, theft, misuse, diversion, or Biosafety and biosecurity encompass the intersection of bioengineering and biotechnology, along with the evaluation and control of risks to human, animal health, and the environment. This link encompasses the danger created by research and its use, as well as research and application to reduce risk via bioengineering and biotechnology. High-level biosafety laboratories provide a secure and reliable setting, integrating robust containment measures, well-educated staff, and precise biosafety protocols to safeguard scientists from infections while working with microbial pathogens. Simultaneously, they prevent the accidental release of these pathogens into the surrounding environment. In recent decades, there has been the construction and operation of labs with different tiers of protection, the establishment of legal regulations and a laboratory biosafety management system, and these functional labs have played a crucial role in addressing newly emerging and recurring infectious diseases. This review article offers useful insights into the complicated environment of biosafety and biosecurity at both the national and international levels. It advocates for more international cooperation among states, organizations, and stakeholders to ensure the effective implementation of biosafety and biosecurity measures, fostering a safer and more secure global biotechnology landscape. Abstract


Introduction
Biosafety involves principles, techniques, and protocols designed to prevent accidental exposure to diseases and toxic substances, as well as to avoid their unintentional release into the environment.It is not solely an labresponsibility but a collective effort aimed at safeguarding a clean and secure environment (1).The term "biosecurity" can be quite intricate as its meaning can vary in different contexts.According to the guidelines set by the World Health Organization (WHO) (2), biosecurity involves the processes that establish and uphold security and control over dangerous microorganisms, toxins, and related materials.Laboratory biosecurity, conversely, centers on safeguarding, regulating, and responsibly managing crucial biological materials within a laboratory to prevent unauthorized access, loss, theft, misuse, diversion, or Biosafety and biosecurity encompass the intersection of bioengineering and biotechnology, along with the evaluation and control of risks to human, animal health, and the environment.This link encompasses the danger created by research and its use, as well as research and application to reduce risk via bioengineering and biotechnology.High-level biosafety laboratories provide a secure and reliable setting, integrating robust containment measures, well-educated staff, and precise biosafety protocols to safeguard scientists from infections while working with microbial pathogens.Simultaneously, they prevent the accidental release of these pathogens

National Frameworks and Guidelines
Many developing countries have utilized developedcountry principles as a starting point and then modified or amended them to conform to local legislation and situations.At the international level, United Nations Industrial Development Organization (UNIDO), Food and Agriculture Organization (FAO), World Health Organization (WHO), United Nations Environment Program (UNEP), and other organizations have developed guidelines that have assisted poor nations in building their own biosafety guidelines (9).In Pakistan, a national biosafety strategy to control all biotechnology development in the country is urgently needed to address the problems (10).Enacting this strategy has the potential to enhance biotechnology research and development in Pakistan and facilitate cooperative relationships amongst native and international researchers and analytical facilities interested in collaborative research, experimentation, or on-site evaluations involving GMOs.
The main goal is to underline the creation of the following Biosafety Guidelines.The framework, developed by the National Biosafety Committee (NBC), are not designed to impose strict mandate that impede pursuits related to the advancement of recombinant DNA technology within the country (11).However, it's important to strike a balance, avoiding excessive permissiveness that might inadvertently encourage negligence and irresponsible behavior among certain researchers or laboratories (12).These guidelines cover a broad spectrum of activities involving gene manipulation through recombinant DNA technology, without regard to their specific goals.This encompasses tasks like creating biotechnologically modified plants, animals, and microorganisms, producing vaccines, manufacturing Genetically Modified Organisms (GMOs) for industrial purposes and their resulting products, and introducing GMOs into the setting for pilot studies and mercantile usages (13).The National Biosafety Committee acknowledges that these regulations are not exhaustive and anticipates that further modifications and revisions will be necessary in the future (14).As new information becomes accessible, it will be incorporated to enhance the current criteria, making them more pragmatic and supportive of the country's broader biotechnology advancement (15).

Basis of Biosafety Guidlines
The biosafety regulations aim to safeguard human health and the environment by minimizing the potential harm that could arise from laboratory activities involving recombinant DNA and the intentional venting of resulting Genetically Modified Organisms (GMOs) and their derived yields (16).Under these guidelines, the term "regulated material" comprises of all genetically altered things/ essentialities, including DNA and RNA medicinal, viroid's, viruses, cells, and other beings that have been genetically altered or manipulated.It also includes their derivatives and any waste or by-products resulting from genetic engineering procedures, whether or not they contain viable organisms (17).These Guidelines are organized into two sections.The first section addresses the handling of regulated activities in laboratories and outdoor settings.The second section outlines the approval processes necessary to declassify controlled materials, enabling their unrestricted distribution and commercial utilization.
Three stages of monitoring and implementation are envisaged in the proposed guidelines.First and foremost, the Principal empiricist and experiment or are accountable to themselves and the community (19).The Institutional Biosafety Committee (IBC) will oversee monitoring and inspection.The IBC is a key organ that plays a pivotal role in the overall setup.It serves as the foundation for the entire setup (20).The Committee will accept submissions, make recommendations for laboratory setup and scheduled releases, and adequately get them surveilled.All facts and findings that must be sent to the following two layers must go through the IBC, which must therefore be comprised of persons who are skilled to comprehend the potential dangers linked with each, and assess their respective significance.(21).Secondly, a Technical Advisory Committee (TAC) will be responsible for the technical review of all licensing applications, ensuring that GMOs or any related products being considered have undergone a comprehensive risk assessment in accordance with these established criteria (22).Third, the Ministry of Environment will establish a National Biosafety Committee (NBC) to serve as the overseeing authority responsible for thorough oversight, control of hazards, and the market introduction of all governed products.(11).

Laboratory Work
These rules apply to all investigative endeavors, regardless of where they take place, be it in educational or research facilities, developmental organizations, or private enterprises, that are related to the utilization and application of GMOs and their derivatives.To ascertain whether a particular project falls within the purview of these regulations, one should prepare a proposal and submit it to the Institutional Biosafety Committee (IBC) (23).In cases where there is no adequately established Institutional Biosafety Committee (IBC) or if one is unavailable, the identical proposal should be presented to the Technical Advisory Committee (TAC) for assessment through the supervising ministry (24).
Researchers who possess any doubts or questions regarding these issues are advised to pursue advice through relevant directorial instigation or contact the Ministry of Environment in Islamabad (25).

Field work
These biosafety standards encompass all facets of field testing for genetically modified plants, animals, and microorganisms.Typically, all field ordeals should follow prior laboratory research.It is a customary practice that genetically modified organisms developed in the laboratory undergo field testing before any Introduction of products into the ecosystem for public use or distribution (26).The suggested field trials should include the following:  Redo the trials conducted in the lab and verify the results obtained from laboratory examinations (27). Collect exact and reliable findings concerning the stability of genetically modified traits, their manifestation, and the passing on of these traits through generations in real-world conditions (28). Penalize the field credibility of genetically modified organisms, including factors such as their survival, reproduction, and competitive abilities (29). Assess the capacity of genetically modified organisms to adapt or evolve in response to varying environmental conditions (30). Assess the comprehensive environmental influence (31).

Commercial Release of Regulated Materials
These rules also describe the steps required to commercially deregulate controlled materials.As a standard practice, it is necessary to furnish comprehensive information from field trials to assess Journal Homepage: www.jwmdc.comwhether the genetically modified organism or its resulting product is eligible for public approval or authorization.In this context, it is crucial to present all the information, factors, and data that formed the basis for the approval of commercial release in a different country (32,33).

Guidelines for laboratory work
Four distinct biosafety levels are defined, each involving a combination of laboratory protocols and methodologies, safety gear, and laboratory infrastructure tailored to the specific tasks performed, the risk associated with the infectious agents, and the purpose or function of the laboratory (34).

Biosafety Level 1
Biosafety Level 1 practices, equipment, and facilities are appropriate for use in educational training labs at the undergraduate and secondary school levels, as well as in other facilities where the work involves wellcharacterized strains of live microorganisms that are not known to cause illness in healthy adults (35).Some examples of bacteria that meet these criteria are Bacillus subtilis, Naegleria gruberi, and the infectious canine hepatitis virus.It's important to note that many agents, while typically not associated with human disease, can still pose a risk to specific vulnerable populations such as young individuals, the elderly, or those with weakened immune systems (36).Therefore, assuming that vaccine strains, even if they have undergone multiple in vivo passages, are entirely non-virulent would be a mistake (37).

Biosafety Level 2
Biosafety Level 2 techniques, equipment, and facilities are suited to clinical, diagnostic, educational, and other facilities that operate with a wide range of indigenous moderate-risk agents found in the population and linked to varied degrees of human disease (38).Microorganisms classified at this containment level can be safely manipulated on an open laboratory bench using appropriate microbiological techniques, provided that the risk of generating aerosols is low.Some microorganisms falling under this category include the Hepatitis B virus, salmonellae, and Toxoplasma spp.Personnel working with these agents should be cautious about potential risks, including accidental self-inoculation, ingestion, and exposure of the skin or mucous membranes to infectious materials (39).Procedures with a significant potential for generating aerosols that could expose personnel should be conducted within primary containment equipment or devices (35).

Biosafety Level 3
Biosafety Level 3 practices, equipment, and facilities are employed in clinical, diagnostic, teaching, research, or production facilities where activities involve indigenous or exotic agents, and there is a tangible risk of infection through aerosols.In such cases, the diseases associated with these agents can be severe or even fatal (35).

Biosafety Level 4
Dealing with dangerous and uncommon materials that carry a substantial personal risk of causing lifethreatening illnesses requires the utilization of Biosafety Level 4 procedures, safety gear, and facilities.Any activities involving potentially infectious diagnostic substances, isolated specimens, or animals infected naturally or through artificial means can potentially expose and infect laboratory staff.An illustration of a microorganism classified under Level 4 is the Lassa fever virus (34,41).

Genetically modified microorganisms
When conducting fieldwork involving genetically modified microorganisms, it is essential to initially evaluate the characteristics of the biological system in question ( 42), This assessment can be summarized as follows:  If the microorganisms have a track record of safe use in field settings, the work can proceed following the established standards for that particular microorganism (43, 44)  These microorganisms originate from strains that have been previously documented in fieldwork. They perform the same functions as strains used in previously documented fieldwork.The work is limited to locations and environments Similar to past environmental circumstances and presumed to have a track record of being safely utilized.If the specified criteria are not met by the experimental microorganisms, the work can advance within suitable levels of containment (45).The proposed containment methods must meet one or more of the following criteria:  Appropriate biological confinement occurs when Microorganisms are made non-replicable before conducting tests in the field; or Alterations are implemented to restrict the persistence of microorganisms beyond designated areas (46). Only in a specific location may genetically inserts and structures be swapped or transferred to other bacteria (47,48,49). There are physical systems established to contain microorganisms within the designated target areas or trial sites (50).When dealing with microorganisms that have not previously been field-tested safely, the research can commence with the primary evaluation of risks to appraise the complete spectrum of plausible environmental consequences.Microbes identified as "problematic" in this assessment (51,52) and engineered for the,  Supporting the growth and nutrition of plant species, potentially providing an abundance of nutrients that could change the chemical composition of neighboring plants. Eliminating detrimental substances could lead to additional damage. Utilizing biological methods to manage plant pests that might outnumber the intended species, consequently generating harmful or diseasecausing substances that can spread within the wild populations at the trial location.

Containment measures must meet following requirements:
o There is an absence of intermingling or crossbreeding occurring (60).o Precautions have been put in place to restrict the dissemination of plants and plant materials (61).o The expression of the introduced gene remains consistent and does not alter in response to shifting environmental conditions (62).In the event of plants that have not previously been used safely in the experimentation (63,64), the initiation of work can start by conducting an initial risk evaluation to establish, assess, and ascertain potential risks:  The experimental site's ecological effects. Increased disease and pest resistance. A proclivity for gauntness. The impacts on other organisms, including both intended and unintended subjects.

Genetically modified animals
The following guidelines (65, 66, 67) should be followed when breeding genetically engineered animals: a. Genetically engineered animals must be raised in distinct breeding facilities that are easily identifiable as separate from non-engineered animals.Furthermore, genetically modified animals should be housed in separate facilities.If it becomes unfeasible to house them individually, small laboratory animals can be grouped for breeding purposes.

International frameworks and guidelines
It draws attention to the biosafety and biosecurity training obligations imposed on states parties by three international regulatory frameworks (68).Regarding each regime, the obligation to give evidence of conformity by reporting the existence of such provisions is reviewed.It claims that these technologies can be used to create and distribute complementary teaching and training resources for biosafety and biosecurity (69,70,71).It highlights how these building blocks can help life and related scientists better carry out their duty to protect their research from future exploitation by increasing their grasp of biosecurity risks (72,73).It is undeniable that "global health governance" has emerged as a prominent concept on the global stage over the last 10-15 years, alongside the twin concepts of globalization and global governance (74).The World Health Organization (WHO) and the United Nations (UNAIDS, UNFPA, UNICEF, and the UN Economic and Social Council, among others) have both done work that has highlighted the importance of international cooperation and a common approach to global health oversight and monitoring (75,76).By considering the World Health  (77).Combining responses to all three regimes by addressing biosafety and biosecurity together is of utmost importance.This cross-regime approach helps establish the education and training requirements for States Parties, both presently and in the future.Moreover, it fosters better compliance with all three regimes and, significantly, streamlines the reporting process (68).

International Health Regulations (IHRs)
The WHO International Health Regulations ( 73) have the primary goal of "preventing, mitigating, controlling, and responding to the international transmission of diseases" (78,79).These regulations impose new responsibilities on State Parties, which extend to both domestic and foreign territories, covering a broad spectrum of public health scenarios that go beyond just infectious disease threats (80). Is there an organization or individual responsible for inspecting laboratories to ensure compliance with biosafety requirements, which may include certifying biosafety equipment? Are biosafety procedures put into practice and subject to regular monitoring? Has a bio-risk assessment been carried out in laboratories to inform and update biosafety regulations, procedures, and protocols, including those related to decontamination and infectious waste management? Have diagnostic laboratories received official designation and authorization, or certification at Biosafety Level 2 (BSL-2) or higher, as needed for the relevant tiers of the healthcare system? Have national experiences and findings concerning biosafety been assessed, and have reports been shared with the global community?

United Nations Security Council Resolution (UNSCR)
Each of these sectors will require the necessary collaboration between various ministries, which will be encouraged and facilitated.On April 28, 2004, the United Nations Security Council (UNSCR) unanimously adopted Resolution 1540 (69,84).States Parties acknowledge an obligation to abstain from the following: Engaging in actions outlined in Chapter VII of the United Nations (UN) Charter, and in accordance with UNSCR 1540, States Parties commit to refraining from:  Supplying any form of assistance to non-state actors endeavoring to create, acquire, manufacture, own, transfer, or utilize nuclear, chemical, or biological weapons or their delivery mechanisms; (85) And that:  All nations must establish and enforce appropriate, efficient laws that forbid any non-governmental entity from producing, acquiring, owning, developing, transporting, transferring, or utilizing  nuclear, chemical, or biological weapons, along with their delivery systems, especially for terrorist objectives.This should also encompass any efforts to partake in such activities, aid, or provide financial support for them (86)  All countries should put into effect and uphold sufficient domestic regulations to inhibit the proliferation of nuclear, chemical, or biological weapons and their delivery systems, which should encompass proper oversight of related materials (69, 87).

The Biological Weapons Convention (BWC)
Under this Convention (83,84), any State Party commits to refraining from creating, manufacturing, storing, or acquiring (88,89):  Microbial or other biological agents, as well as toxins, irrespective of their origin. Weapons, machinery, or means of delivery intended for deploying these substances or toxins for aggressive purposes or in warfare.This includes production techniques, types, and quantities that lack a valid rationale for prevention, protection, or other peaceful intentions.

Implementation of biosecurity and biosecurity measures
Biosafety involves the adoption of laboratory protocols and processes, the incorporation of particular structural aspects in laboratory facilities, the utilization of safety gear, and the establishment of suitable occupational health programs when dealing with potentially infectious microorganisms and other biological risks (69).These precautions are designed to safeguard laboratory personnel, the public, agriculture, and the environment from potentially harmful agents and other biological hazards.In recent years, there has been increased attention on laboratory-acquired illnesses (LAIs), particularly in laboratories with high (biosafety level 3, or BSL-3) and maximum (BSL-4) containment.LAIs can occur in various types of labs, including research facilities, clinical laboratories, or animal research centers, and it can be challenging to determine whether the infection originated in the lab or the broader population.LAIs also pose a significant public health concern, as an infected lab worker can transmit the infectious disease to colleagues, family members, or the wider community (90)

Challenges
Biosafety has become an increasingly important aspect of worldwide security, with broad-reaching consequences that impact various fields, such as healthcare, agriculture, scientific research, technology, education, and the armed forces.It constitutes a vital element of national security and deals with concerns such as concealment, rapid dissemination, accidental release, overflow, and the potentially devastating consequences of biological threats (91).

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Journal Homepage: www.jwmdc.compractices in the life sciences.The core objectives of this network are to raise awareness about both the potential advantages and risks associated with biotechnological progress, enhance education in responsible life sciences, and advocate for effective policies that ensure sustainable development in biotechnology.Notably, the INB is currently developing a digital platform that will enable the access, download, upload, and sharing of customizable educational and training materials, including technical briefings, case study videos, scenario-based exercises, and immersive learning experiences like virtual reality laboratory tours.The growing global need for improved disease diagnosis and control has led to an expansion of diagnostic and research capabilities.However, this increased capacity for detecting infectious diseases hasn't always been accompanied by a proportional enhancement of biosafety and biosecurity capabilities, especially in resourceconstrained nations.Various obstacles impede the establishment or expansion of sustainable management capabilities for biosafety and biosecurity in these countries (92).Despite of the presence of international initiatives and national regulations, concerns related to biosafety and biosecurity continue to exist.This underscores the necessity for enhanced cooperation between the scientific and policy sectors to foster a culture of safety and security within the bioscience community (93).In 2003, there were instances of laboratory-acquired SARS infections that took place within biosafety level 3 (BSL3) and BSL4 (the highest containment level) laboratories (94).As per a WHO investigation, these infections resulted from poor program management, including inadequate laboratory procedures and insufficient training.Likewise, even the most robust security measures can be compromised if the individuals with access to hazardous infections lack trustworthiness, reliability, and adherence to security protocols (72).Since 2003, there have been more than 100 biosafety and biosecurity incidents at laboratories in the United States, according to the Associated Press (95).Texas A&M University, for example, was fined $1 million USD and had to cease all of its select agent research due to failures to adequately and accurately record events (96).Pirbright Laboratory in the United Kingdom unintentionally released foot-and-mouth disease virus into the neighborhood due to a leak in its wastewater pipes, which were known to be in need of maintenance (97, 98, and 99).

Conclusion and Future Perspective
The study underscores the significance of national and international frameworks and norms in promoting the safe and secure handling of biological materials.These regulations play a crucial role in preventing accidents and mitigating risks associated with biological research and operations.Given the dynamic nature of biotechnology and emerging biological threats, continuous adaptation and global collaboration are essential.Adhering to these principles allows us to strike a balance between harnessing the benefits of biotechnology and minimizing potential threats to both people and the environment.The trajectory of future biosecurity and biodefense initiatives may be influenced significantly by the occurrence of the "next event."Nevertheless, the most likely scenarios involve events that unfold inadvertently and spontaneously, posing threats to human and animal health.These threats may emerge due to the development of new diseases or the resurgence of existing ones in response to changing environmental or cultural factors (100).In light of the substantial threats posed by evolving infectious diseases and bioterrorism, the responsibility for workplace biosafety and biosecurity extends to all individuals.However, the primary duty lies with governments globally, necessitating heightened awareness and readiness to detect and contain hazardous biological agents.This responsibility transcends the establishment of a safe environment solely for laboratory workers; it also involves the adoption of biosafety commitments.The well-being of laboratory personnel, the proper handling of pathogens, and the overall laboratory environment hinge on the implementation of secure and efficient laboratory procedures and pathogen management.These factors are pivotal in ensuring the successful completion of laboratory experiments.Acknowledgement: The author would like to acknowledge National University of Medical Science (NUMS), Islamabad, Pakistan and Kohat university of Science and Technology (KUST), Khyber Pakhtunkhwa, Pakistan.

Figure 1 : 2 . 2
Figure 1: Basic elements providing for implementation of biosafety regulations

Figure 2 :
Figure 2: Biosafety levels (BSL) 1 to 4 As a globally applicable legal framework, the International Health Regulations of 2005 are binding on all 194 States Parties around the world, encompassing all member states of the WHO (81).The WHO questionnaire dispatched to States Parties in 2011 for the evaluation of their national implementation of the International Health Regulations (IHRs) is organized into 13 sections (82), each of the 13 sections in the WHO questionnaire addresses the eight essential capacities mandated for States Parties, which include coordination, legislation policy, surveillance, response, preparedness, laboratory, and human resource capacity.It also encompasses four specific hazard categories, which are zoonotic events, chemical events, food safety, and radiation emergencies, and concludes with points of entry (83).It also contains the following inquiries (84):  Do individual laboratories have access to biosafety guidelines? Are there regulations, policies, or strategies in place for laboratory biosafety? Has an entity been assigned responsibility for laboratory biosafety and biosecurity? Have biosafety guidelines, manuals, or Standard Operating Procedures (SOPs) been distributed to laboratories? Are the staff involved adequately trained in  biosafety guidelines? Has the categorization of microorganisms by risk group at the national level been completed?Journal Homepage: www.jwmdc.com

Figure 3 :
Figure 3: Analysis, designing and the steps of implementation of biosecurity Autoinoculation and ingestion are also major risks to those who work with these substances.Mycobacterium tuberculosis, St. Louis encephalitis virus, and Coxiella burnetii are examples of such agents for which Biosafety Level 3 protections are widely suggested (40).

Table 1 :
(73)ddress biosecurity concerns, a rising number of study resources for biosecurity-related educational training have been developed and made available(73).For instance, the United Nations Interregional Crime and Justice Research Institute has founded the International Network on Biotechnology (INB), a worldwide coalition that includes academic research institutions, nongovernmental organizations, international entities, and other stakeholders interested in promoting responsible Biosafety and biosecurity represent crucial foundations of global health security and are fundamental aspects of preventing the spread of biological weapons.