Biological Safety

Safety for people, animals & plants

Biosafety and Biosecurity Training

“A safe, healthful, and secure environment for scholarship and research.”

Vinod Jyothikumar,PhD

Executive Summary

2 Biological Agents Overview

3 What are Biosafety Materials ?

4 Understanding Biosafety

Materials

5 Classification of Biosafety

Levels

1 Introduction

Biosafety Biosecurity &

Bioterrorism

7 High Containment Biodefense

Research Laboratories

9 Testing Biological Agents

11 Research Aspects of

Bioterrorism

12 The standard code of Practice for

Biosafety

8 Microbial Forensics Analysis

10 Key Concerns in Biological

Research

Biosafety-Biosecurity Bioterrorism

hot

zones

virus hunters

space suits

Biohazard-Labels

Biosafety-Biosecurity Bioterrorism

An agent of biological origin that has the capacity to produce deleterious effects on humans, i.e. microorganisms, toxins and allergens derived from those organisms; and allergens and toxins derived from higher plants and animals. The Hazard Communication Sign is not intended to restrict access, but to communicate that biohazardous materials may be present in the laboratory.

The term biosecurity has been applied more broadly to encompass efforts to prevent harm from both intentional and unintentional introductions of organisms to human health and infrastructure and the environment, as well as to the agricultural crop and livestock industries.

Biosecurity

The application of knowledge, techniques and equipment to prevent personal, laboratory and environmental exposure to potentially infectious agents or biohazards.

Biosafety

Bioterrorism is a form of terrorism where there is the intentional release of biological agents (bacteria, viruses, or other germs). This is also referred to as germ warfare.

Bioterrorism

Category A agents, as classified by the U.S. Department of Health and Human Services’ (HHS) Centers for Disease Control and Prevention (CDC), are included because they are of the highest concern as potential threats. They have the potential for major public health impact and social disruption and are also known to have been studied by some countries for use in biological warfare.

Category B agents, which are defined as a “second highest priority” by CDC, are also included. Although these agents are fairly easy to disseminate, they generally cause moderate illness and low death rates.

Category C agents, which are considered to be “third highest priority” by CDC, are not included because they are currently not major bioterrorism threats. However, these agents are emerging as infectious disease threats that CDC believes could, in the future, be engineered to produce biological weapons.

Biological Agents Overview

• Any biological material capable of causing harm to humans, animals, plants

• BioHazardous Agents – Pathogens that can replicate & cause disease include:

– Bacteria (Streptococcus pyogenes)

– Fungi (Candida, Histoplasma)

– Viruses (HIV, HBV)

– Parasites (Giardia, Strongyloides)

• Toxins – Microbial poisons

– Exotoxins – produced by bacteria

• Clostridium botulinum - food poisoning, one of most deadly

• Clostridium tetanii – tetanus

• Corynebacterium diphtheriae – diptheria

• Bioterrorism use

– Endotoxins – released from cell wall when bacteria disintegrates

What are Biosafety Materials?

Infectious Diseases

Source:Terrorism and Other Public Health Emergencies: A Reference Guide for Media

Understanding Biosafety

Materials

Infectious diseases are caused by the invasion of the body by harmful microorganisms.

Microorganisms multiply and make the person

sick by attacking organs or cells in the body. These harmful microorganisms include viruses

and bacteria, as well as certain other microscopic organisms, and are sometimes called pathogens.

There is usually a lag time, called an incubation period, between when a person is infected and when the symptoms appear. infectious diseases.

vibrio cholera bacterium

Understanding Biosafety

Materials

People can become infected with these diseases in any number of ways, including consuming contaminated water or food, being bitten by insects or animals, or inhaling or touching the microorganisms or their spores.

-Spores are produced by certain bacteria and plants. Like seeds, spores do not grow until the environment is conducive for them to do so. They are highly resistant to heat and other environmental factors. All of the diseases discussed in this section are considered infectious diseases.

Illnesses caused by chemical agents by contrast, are not.

Contagious Diseases

A contagious disease is an infectious disease that can be “caught” by a person who comes into contact with someone who is infected. Not all infectious diseases are contagious.

Source: Terrorism and Other Public Health Emergencies: A Reference Guide for Media

Understanding Biosafety

Materials

Exposure to a contagious disease usually happens through contact with the infected person’s bodily fluids or secretions, such as a sneeze.

Depending on the disease, the level of contact required to pass on the

illness could be as casual as water droplets in the air from a cough (e.g., smallpox).

The level of contagiousness has nothing to do with how serious the resulting disease may be. For example, pneumonic plague and the common cold are both highly contagious, but pneumonic plague is obviously a much more serious disease.

There are some infectious diseases that

are not contagious at all, no matter how close the contact with an infected person (e.g., botulism, tularemia). Francissella tulareniasis

Classification of Biosafety Levels

Risk Group 1 BSL-1

Agents not associated with diseases in healthy adult humans

Risk Group 2 BSL-2

Agents associated with human disease: rarely serious: preventative of therapeutic resources available

Risk Group 3 BSL-3

Agents associated with serious of lethal human disease: rarely serious: preventative of therapeutic resources available

Risk Group 4 BSL-4

Agents likely to cause serious of lethal human disease: rarely serious: NO preventative of therapeutic resources available

Biosafety level 1 (BSL1) Suitable for working with microbes that pose no risk, or a very limited risk, to personnel or the environment. Work is often conducted on open lab bench tops. No special containment equipment is needed and foot traffic through the lab is not restricted. However, there are limited safety measures. For instance, contaminated work surfaces must be decontaminated; and eating and drinking is not permitted. However, Some organisms may cause disease in immunocompromised individuals.

Biosafety level 2 (BSL2) Built for work with microbes of moderate risk. Access is limited, laboratory coats must be worn, and biosafety signage is required at entrances. In addition, special cabinets that contain and filter the air are used for working with microbes that can move through the air (aerosols). Generally required for any human-derived blood, bodily fluids, tissues in which infectious agent may be unknown

BSL1+BSL2+: Biosafety level 2 facility +

biosafety level 3 working techniques

Classification of Biosafety Levels

(transmission electron micrograph of E.

coli)

(transmission electron micrograph of

hepatitis B virus)

Biosafety level 3 (BSL3) Used for work with microbes that can cause serious injury or, in cases of an extreme exposure, death. Agents with potential for respiratory transmission, may cause serious and potentially lethal infection In addition to the safety measures of level 2 labs, there are several other safeguards. People younger than 16 are not allowed in these labs; doors must be kept closed; windows are sealed. Surgical gloves and masks, or respirators, are used. The rooms are kept at a lower air pressure level than surrounding rooms, and the outdoor environment to contain aerosols. Air in the room also is continuously filtered through HEPA filters that are able to catch minute particles. These filters are regularly disinfected and changed. May be studied at BSL-2 for diagnosis

Classification of Biosafety Levels

BSL3+: Biosafety level 3 facility + biosafety level 4 working

techniques

(F. tularensis under

direct fluorescent

antibody stain)

Biosafety level 4 (BSL4) Highest level of safety measures to protect those working inside them and in adjacent areas. Used for work with highly infectious or dangerous microbes. Dangerous and exotic agents with high risk of life-threatening disease, aerosol-transmitted Contain the safety measures of BSL3 facilities. In addition, access is very restricted. Entry logs are kept for the labs. Lab workers must wear protective suits and undergo a disinfectant shower upon leaving. The labs have double-door airlocks. Special nonbreakable containers are used for working with the microbes. Materials in the lab are destroyed or decontaminated before leaving. Strictly controlled access to the laboratory; Changing clothing before entering and exiting

lab (showering upon exiting recommended) Decontaminating all material exiting facility

Classification of Biosafety Levels

(transmission electron

micrograph of Ebola

virus)

• BSL-1: high schools, community colleges, municipal drinking water treatment facilities

• BSL-2: local health departments, universities, state laboratories,

private laboratories (hospitals, health care systems), industrial laboratories (clinical diagnostic companies)

• BSL-3: state health departments, universities, private companies,

industry, federal government (NIH, CDC) • BSL-4: only 15 facilities in the US

– 9 federal (CDC, NIH), 4 university (Georgia State University, University of Texas Medical Branch), 1 state, 1 private

– Renovations underway at several labs, new facilities proposed at additional sites

Classification of Biosafety Levels

BSL-3 Labs Registered with the CDC and USDA Select Agent Program, by Sector

High Containment Biodefense

Research Laboratories

Gigi Kwik Gronvall, PhD, is a Senior Associate; Joe Fitzgerald, MPH, is a Senior Associate; Allison Chamberlain is an Analyst; Thomas V. Inglesby, MD, is COO and Deputy Director; and Tara O’Toole, MD, MPH, is CEO and Director; all are at the Center for Biosecurity of the University of Pittsburgh Medical Center, Baltimore, Maryland.

High Containment Biodefense

Research Laboratories

What We Don’t Know About Biological

Weapons Of Mass Destruction

Medical experts do not know if bacteria have been engineered to be resistant to antibiotics.

Experts do not know how potent the strains will be in a biological attack.

Experts do not know if the illnesses caused by some of these agents will be immediately recognizable.

There is disagreement on how long some of these agents can survive in the environment.

Experts do not know if the diseases these agents cause will be the same as past epidemics (e.g., is today’s smallpox the same as the disease that was eradicated?).

A hypothetical sample microbial forensic analysis flowchart for characterizing evidence from an overt attack.

Microbial Forensic Analysis

Source: Bruce Budowle, FBI Laboratory

Testing for Biological Agents

Quick diagnosis and treatment of a patient exposed to a biological agent are key to saving that patient’s life. The following list provides descriptions of the types of tests that may be run

•This is a examining a building or an area for the presence of a biological or chemical agent. Testing is usually a two-step process. The confirmation test can take up to 72 hours depending on how fast the bacteria grow and can be positively identified.

Environmental testing

•Nasal swab is an environmental test that may be used to assist in a public health investigation to determine the presence of a bioterrorism agent in an area or building. Nasal swabs are not used to determine if a person is infected by a given agent.

nasal swab

•A culture is a method for growing an organism in the lab (for example, in a Petri dish). A culture can help in identifying a bacteria or virus. Cultures are the most readily available technology for clinical diagnosis of a suspected case of bioterrorism but can be time consuming to complete

Culture

Testing for Biological Agents

•A blood culture is a test that looks for bacteria or viruses in the blood. Blood is drawn from a patient, and the sample is sent to a special laboratory for analysis. The sample is incubated, or kept in a warm place, at a certain temperature, and in this controlled environment, the bacteria grow and are isolated from the blood for easier identification.

Blood culture

•“Media” are the nutrient mixes used to grow organisms in a lab. Selective media are used to identify an organism by giving it or depriving it of certain nutrients. For example, selective agents can be added or removed, which “poison” some organisms while allowing others to grow.

Selective media

•Gram staining uses dyes to make a bacterial cell stand out for identification. A specimen is put on a slide and a four-part staining procedure begins. This test may produce results in less than an hour, but it is not specific enough to definitely identify the organism, and a longer confirmation test is still needed.

Gram staining

Testing for Biological Agents

With advanced techniques

•Which look for specific antigens or antibodies and are useful in detecting the presence of toxins. However, antibody production for identification can take time.

Immunoassays

•Such as a polymerase chain reaction, which look at the DNA or RNA to identify an agent. However, sample preparation can take a long time.

Gene amplification

In general, detection and identification using any of these methods is dependent on the sample quantity and quality and the exactness of the processing. A combination of tests will yield the most accurate results. In the absence of immediate results, physicians who suspect bioterrorism may begin a preliminary course of treatment until the lab results are in.

Key Concerns in Biological

Research

The cloning from the new and unknown microorganisms relating to

human health effect

The cloning of toxin gene with LD50 less than 100 ng/kg body weight

The deliberate transfer of drug by resistance to microorganisms different

methods from natural acquisition

The cloning from highly dangerous pathogens

Research Aspects of

Bio-terrorism

• Biosafety in Microbiological and Biomedical Laboratories (BMBL) – U.S. Department of Health & Human Services

– Public Health Service

– Centers for Disease Control & Prevention & the National Institutes of Health

– http://www.cdc.gov/od/ohs/biosfty/bmbl5/bmbl5toc.htm

• Establishes criteria for: – Biological Risk Assessment

– Principles of BioSafety

– Laboratory BioSafety Level (BSL) Criteria

– Laboratory BioSecurity Criteria

– BioHazard Containment

– Decontamination & Disinfection

– Transportation of BioHazards & Infectious Materials

– Select Agents & Toxins…

The Standard Code of Practice for

Biosafety

Partenering and Cooperation

National Biosafety Strategy

Leg

isla

tive

(Reg

ula

tions)

Guid

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es

Educa

tion

& T

rain

ing

Bio

safe

ty

spec

iali

sts

Raising awareness

Preparedness for intentional

release: Pathogen Control

Developing risk assessment

methodologies

Capacity- building for risk

management

Promoting Health Security

1. US Department of Health and Human Services, Centers for Disease Control and Prevention and National Institutes of Health. Biosafety in Microbiological and Biomedical Laboratories. 5th ed. Washington, DC: US Government Printing Office; 2007. http://www.cdc.gov/od/ohs/ biosfty/bmbl5/bmbl5toc.htm. Accessed February 6, 2008.

2. Clemson University Environmental Health and Safety. Biological Safety [online training]. http://ehs.clemson.edu/training/biosafety/index.html. Accessed February 6, 2008.

3. Centers for Disease Control and Prevention. Laboratory-Acquired West Nile Virus Infections — United States, 2002. MMWR Morb Mort Wkly Rep. 2002; 51:1133-1135. http://www.cdc.gov/mmwr/preview/ mmwrhtml/mm5150a2.htm. Accessed February 6, 2008.

4. United States Government Accountability Office. High-Containment Biosafety Laboratories: Preliminary Observations on the Oversight of the Proliferation of BSL-3 and BSL-4 Laboratories in the United States. Publication GAO-08-108T. http://www.gao.gov/docsearch/ abstract.php?rptno=GAO-08-108T. Published October 4, 2007.

References