Special Hazards in the Laboratory

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

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Advanced Laboratory Safety TrainingPart 5 of Texas A & M Laboratory Safety Training Programs

Key Concepts

• Hazardous Biological Substances (Biohazards)• Bloodborne Pathogens• Universal Precautions• Exposure Control Plan• Biosafety in Microbiological and Biomedical Laboratories

(BMBL)• Biological Risk Assessment• BioSafety Level (BSL)Criteria• BioSecurity Criteria• Select Agents & Toxins• BioHazard Containment, Decontamination & Disinfection• TAMU Biological Safety Office

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Why Is Biological Safety Important?

• Use of hazardous biological materials on campus including– Biohazardous agents and bloodborne

pathogens• Bloodborne Pathogens• Prevent disease in personnel• Prevent release of organisms/agents into

environment

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What are BioHazardous Materials?• 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)– Prions (CJD)– Parasites (Giardia, Strongyloides)

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What are BioHazardous Materials?

• Toxins – Microbial poisons– Exotoxins – Produced by bacteria

• Clostridium botulinum - food poisoning, one of most deadly• Clostridium tetanii – tetanus• Corynebacterium diphtheriae – diphtheria• Bioterrorism use

– Endotoxins – released from cell wall when bacteria disintegrates

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Classification System For Biohazardous Materials

BioSafety Levels (BSL)Risk Group 1

BSL 1

• Agents not associated with disease in healthy adult humans

Risk Group 2BSL 2

• Agents associated with human disease; rarely serious; preventative or therapeutic resources available

Risk Group 3 BSL 3

• Agents associated with serious or lethal human disease; preventative or therapeutic resources available

Risk Group 4 BSL 4

• Agents likely to cause serious or lethal human disease; no preventative or therapeutic resources available

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The Standard Code of Practice for BioSafety• 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

• Established criteria– 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

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Researcher Responsibilities

• Files a Biological Usage Authorization (BUA)• Trains all personnel• Establishes emergency procedures• Reports incidents with biologicals• Posts biohazard warning signs• Cooperates with Biological Safety Officer during

inspection visits• For more information covering responsibilities of TAMU

PI’s, visit http://rcb.tamu.edu/

Faculty or Principle Investigator (PI)

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BioSafety Officer

• Dr. Bruce M. Whitney – Institutional Biological Safety Officer– Office of Biological Safety– Texas A&M University

• Tel: 979-458-0683 or 436-1307• Mailstop: 1112 TAMU• Email: brucewhitney@tamu.edu• Email: bwhitney@vprmail.tamu.edu

TAMU Office of Research Compliance

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Institutional BioSafety Committee (IBC)

• Mandated by National Institutes of Health (NIH)• Must review and approve all usage of Risk Group 2 or

higher agents• Must review and approve rDNA usage, formation of

transgenic animals, experiments involving human gene therapy

• Applies to TAMU– Employees & students conducting research involving

infectious biohazards – Research subjects, human or animal

• TAMU IBC: http://rcb.tamu.edu/

TAMU Office of Research Compliance

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Biological Usage Authorization (BUA)

• Describes the Principle Investigator’s research project

• Required when biohazardous material is stored, used, or transported

• Good for 3 years• Can be amended

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Forbidden Agents

• Reference material can be kept in inactive state– (frozen or lyophilized)

• List of all references must be given to IBC

Cannot be brought on

campus!

• Mycobacterium tuberculosis• Hantavirus• Foreign Animal Pathogens• HIV 1 & 2

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CDC Select Agents

• Cannot be brought on campus without prior approval & registration

• Must be registered with the University BioSafety Officer

• Required under• U.S. BioTerrorism Prevention Act• USA Patriot Act (USAPA)• TAMUS Regulations, TEES & TAMU Rules

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The BioSafety Inspection

What is inspected

• Personal Protective Equipment (PPE)

• Training• Signage• Adherence to protocol• Technique• Physical facility• General safety• Risk assessment• Controls• Project Safety Analysis

(PSA)14

Working Safely with Bloodborne Pathogens

What you

should know

• Cleaning• Decontamination• Regulated waste disposal• Pre and post exposure

procedures• Universal Precautions (CDC)• Special hazards

• HIV, HBV research

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What are Bloodborne Pathogens?

• Microbial agents transmitted by the blood and bodily fluids of infected individuals

• Control of Occupational Exposures to BloodbornePathogens

• OSHA standard 29 CFR 1910.1030

http://www.osha.gov/SLTC/bloodbornepathogens/index.html

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Exposure Control Plan

• All laboratory supervisors, faculty and principle investigators are responsible for insuring their employees and students comply with the Exposure Control Plan to reduce the risk of occupational exposures to bloodborne pathogens

• All personnel that have the potential to be exposed to bloodborne pathogens MUST be trained to this plan

• A copy of the Exposure Control Plan must be readily available in each work area or laboratory where bloodborne pathogens exposures exist

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Where to Get More Information

• Workers in many different occupations are at risk of exposure to bloodborne pathogens, including Hepatitis B, Hepatitis C, and HIV/AIDS

• In 1991, OSHA issued the Bloodborne Pathogens Standard to protect workers from this risk

• In 2001, in response to the Needle-stick Safety and Prevention Act, OSHA revised the Bloodborne Pathogens Standard, 29 CFR 1910.1030

• The revised standard clarifies the need for employers to select safer needle devices and to involve employees in identifying and choosing these devices

• The updated standard also requires employers to maintain a log of injuries from contaminated sharps

www.osha.gov/SLTC/bloodbornepathogens/index.html

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Where to Get More Training

• Prevention of Exposures to Bloodborne Pathogenshttp://rcb.tamu.edu/bohp/training-1/training

– This training presentation will aid in increasing awareness of the risk of exposure to bloodbornepathogens and techniques and practices for preventing exposure and abating hazards in Engineering laboratories and facilities

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Nano Technology Safety in the Laboratory

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

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Key Concepts

• Nanotechnology• Nanoparticle Safety

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Nanometer-Scale Particles & NanoTechnology

• A nanometer (nm) is one-billionth of a meter– Or 1 x 10-9 m – or one millionth of a

millimeter

Sheet of Paper• 100,000 nm thick

Our DNA• 2 to 12 nm in diamete

Red Blood Cells• 5,000 nm in diameter

Human Hair• 10,000 to 50,000 nm

Virus• 10 to 100 nm in dimension

For Comparison

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What is Nanotechnology?• Nanotechnology is somewhat loosely

defined, although in general terms it covers engineered structures, devices and systems that have a length scale of 1 to 100 nanometers

• At these length scales, materials begin to exhibit unique properties that affect physical, chemical and biological behavior

• The use of the term “nanotechnology” can be misleading since it is not a single technology or scientific discipline

• Rather it is a multidisciplinary grouping of physical, chemical, biological, engineering, and electronic, processes, materials, applications and concepts in which the defining characteristic is size

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Risks in Nanotechnology

• Existing research shows little evidence to suggest that the exposure of workers arising from the production, handling, and processing of nanoparticles has been adequately assessed

• Current knowledge is inadequate for risk assessment purposes

• No information has been identified about worker exposures to nanoparticles in the university/research sector or in the new nanoparticle companies

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Exposure Controls for Nanotechnology

• Total enclosure of all processes • Storage of all Nano-materials in total enclosure• Local exhaust ventilation, with HEPA filtration• General ventilation• Limitation of numbers of workers, and exclusion of

others• Reduction in periods of exposure, via SOP’s and

personnel training• Regular cleaning of wall and other surfaces;

documented cleaning schedule

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Exposure Controls for Nanotechnology

• Use of appropriate personal protective equipment

• Prohibition of eating and drinking in laboratories and controlled areas

• Transport of Nano-materials within a secondary containment device

• Immediate cleanup of ALL spills & discharges• Collection of all nanoparticle waste materials

for disposal in compliance with the TAMUHazardous Waste Management Plan

• Follow safety procedures outlined in the first Safety in the Laboratory presentation

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PPE for Nanotechnology

• Individuals working with nanoscale materials must wear a HEPA rated respirator to reduce potential health risk

• Free, unfixed, nanoscale materials must be used in an exhausted glove box or similar containment

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Guidelines for Working Safely with Nanoparticles

To receive the “Guideline for Working Safely with Nanotechnology and Nanoparticles”, contact EHS Engineering Safety at 845-2132

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Radiation & Laser Safety in the Laboratory

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

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Key Concepts

• Radiation Safety• Laser Safety

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Ionizing Radiation: Overview• Three types of radiation emitted from various isotopes:

Alpha, Beta, and Gamma• Damage to the body occurs through interaction with a part of the cell

or indirectly by the formation of free radicals and ultimately changes within the DNA of the cell

• The amount of damage depends on many factors including the dose rate, the size of the dose, the site of exposure

• Effects may be short or long term, acute short term effects = 100,000 mrads (100 rad) in less than 1 week will cause nausea, diarrhea, fatigue, hair loss, sterility and easy bruising – Over 600 rads is fatal– Long term low exposures can lead to cancer

• No completely safe limit of exposure is known which is why we try to reduce exposures to ALARA (as low as reasonably achievable) levels

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Ionizing Radiation: Definition of Terms

• Roentgen ( R ) - The amount of x- or gamma radiation that produces ionization resulting in one electrostatic unit of charge in one cc. of dry air at standard conditions

• Roentgen Absorbed Dose (rad) - The mean energy per unit mass imparted by ionizing radiation in a mass – One rad is 100 ergs absorbed per gram or 1 rad = 0.01

Gy• Roentgen Equivalent Man (rem) - A unit of absorbed done

(in rad) times a quality factor used to express the relative biological effect of the particular radiation as compared to gamma radiation – Personal exposure limits are often expressed in rem

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Ionizing Radiation: Definition of Terms

• Sievert (Sv) - Unit of absorbed radiation dose (in Gy) times the quality factor fo the radiation as compared to gamma radiation – Equivalent to the Gy times the quality factor and

equivalent to 100 rem• Gray (Gy) - Unit of absorbed radiation dose equal to one

joule of absorbed energy per kilogram of matter, 1 Gy = 100 rad

• Quality Factor - Weights the absorbed dose for the biological effectiveness of the particles producing the absorbed dose

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Ionizing Radiation: Alpha

Alpha charged particles containing 2 protons, 2 neutrons • Can be emitted by from certain heavy atoms such

as uranium and thorium • Move in straight lines, slowly• Can be stopped by a piece of paper or by skin, but

if inhaled or gets into an open wound can very damaging inside the body

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Ionizing Radiation: Beta

ß - Beta particles electrons emitted with very high energy from many radioisotopes

• Positively charged counterparts of beta particles are called positrons and can be shielded by thin metal foils or ¼ inch plastic

– Examples are tritium, phosphorus-32, and carbon-14

• Can be stopped by the skin • Can cause serious damage to the eyes and to the

skin

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Ionizing Radiation: Gamma

• Gamma rays and x-rays, extremely energetic photons, have no mass or charge

• Gamma rays are emitted from the nucleus during decay, and can be produced by particle accelerators, and nuclear reactors

• X-rays are emitted from the electron shells • Extremely dense materials, such as lead or depleted

uranium are needed to shield these particles• Neutrons

– Uncharged particles, emitted from the nucleus during decay

– Can be shielded by water, paraffin, boron, and concrete

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Ionizing Radiation: Threshold Limit Values (TLVs)

• TLVs are published by the ACGIH (American Conference of Governmental Industrial Hygienists) and are used in many jurisdiction’s occupational exposure limits or guidelines̶ 20 mSv: TLV for average annual dose for radiation workers,

averaged over five years̶ 1 mSv: Recommended annual dose limit for general public -

International Commission on Radiological Protection (ICRP)̶ 1 rem = 20 mSv̶ 1 Sv = 100 rem

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Ionizing Radiation: Risk• The average lifetime risk of death from cancer following

an acute dose equivalent to all body organs of 0.1 Sv (10 rem) is estimated to be 0.8%

• This increase in lifetime risk is about 4% of the current baseline risk of death due to cancer in the United States

• The baseline risk of cancer induction in the United States is approximately 25%

• Another way of stating this risk– A dose of 10 mrem creates a risk of death from cancer of

approximately 1 in 1,000,000

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Lasers & Laser Systems: Class 1• Class 1 Laser

– Not able to cause biological injury, exempt from any controls

– Lasers are considered safe based upon current medical knowledge

– Includes all lasers or laser systems which cannot emit levels of optical radiation above the exposure limits for the eye under any exposure conditions inherent in the design of the laser product

– There may be a more hazardous laser embedded in the enclosure of a Class 1 product, but no harmful radiation can escape the enclosure

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Lasers: Class 2• Class 2a & 2b Lasers

– Ocular hazard only if viewed for more than the blink response (0.25 seconds)

– All wavelengths are of the visible (0.4-0.7m m) portion of the electromagnetic spectrum and less than 1mW of power

– Class 2 laser or laser system must emit a visible laser beam

– Because of its brightness, Class 2 laser light will be too dazzling to stare into for extended periods

– Momentary viewing is not considered hazardous since the upper radiant power limit on this type of device is less than the MPE (Maximum Permissible Exposure) for momentary exposure of 0.25 second or less

• Intentional extended viewing, however, is considered hazardous

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Lasers: Class 3• Class 3 laser or laser system can emit any

wavelength, but cannot produce a diffuse (not mirror-like) reflection hazard unless focused or viewed for extended periods at close range– Class 3 lasers are not considered a fire

hazard or serious skin hazard • Any continuous wave (CW) laser that is not

Class 1 or Class 2 is a Class 3 device if its output power is 0.5 W or less– Since the output beam of such a laser is

definitely hazardous for intra-beam viewing, control measures center on eliminating this possibility

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Lasers: Class 3A & 3B

• Class 3a– Ocular and skin hazard for direct exposures– Beams may include wavelengths of visible, infrared

or ultraviolet ranges– CW’s maximum power between 1mW and 5mW– Pulsed radiant energy cannot exceed 0.125J with

exposure time <0.25s• Class 3b

– Beams may include wavelengths of visible, infrared, or ultraviolet ranges

– Emit no more than 5mW-500mW (or 5W) of CW power or 10 J/cm2 of Pulsed power

– Acute hazard to eyes and skin from direct beam – Diffuse reflections may be a hazard

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Lasers: Class 4

• Class 4 laser or laser system is any that exceeds the output limits Accessible Emission Limits (AEL) of a Class 3 device – As would be expected, these lasers may be

either a fire or skin hazard or a diffuse reflection hazard

– Very stringent control measures are required for a Class 4 laser or laser system

• Class 4 – Ocular and skin hazard to the direct beam

and the diffuse reflections– Beams may include wavelengths of visible,

ultraviolet or infrared ranges– CW power greater than 5W, or pulsed

radiant energy greater than 10J/cm2, possible ignition/fire hazard

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Lasers: Safety for Class 3B and Class 4

• Provide key switch interlock systems to prevent unauthorized use

• Wear laser goggles that match the wavelength of the laser system being used

• Prevent reflection of beams - be aware of windows, smooth surfaces and mirrors, watches, etc. in your lab

• Prevent scattered light emissions with beam stops

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Lasers: Safety for Class 3B and Class 4

• Never align the beam with unprotected eyes • Never intentionally stare into any laser beam • The most important safety measure you can

include in your lab is the proper selection of eyewear̶ Always check with the manufacturer for their suggested

eyewear requirements̶ Look for the wavelengths and Optical Density (OD) values

stamped on the side of the eyewear — does it match the wavelengths of the equipment you are using?

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Lasers: Registration for Class 3B and Class 4

• All Class 3b and Class 4 lasers on TAMU property must be registered with the TAMU Radiation Safety Office

• All Class 3b and Class 4 lasers must be registered within 10 days of arriving on campus

• System Regulation 24.02.02 Health & Safety Program Section on Radiation Safety, Subsection 3.3 on non-ionizing radiation safety, and comply with the Texas Radiation Control Act, Title 25 Texas Administrative Code (TAC) §.289.301, Registration and Radiation Safety Requirements for Lasers (as amended)

• TAMU Radiation Safety Office: 845-0063 or 845-2132

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Lasers: Analysis of Laser Accident Data • 272 accidents reported to Rockwell Laser Industries from 1964 to 1994

reveals that– Eye injury was the most commonly reported laser related accident – Eye injury was involved in slightly over 74% of all of the accidents

recorded– Over 90% of the eye injury cases recorded some functional loss, of

which 77% was permanent

• Statistical data gathered by the Laser Institute of America (LIA) concludes that the most common laser incidents are caused by Nd: Yag (29.7%) and Argon (20.5%) lasers

• More importantly are the occupations which are involved– Out of 12 categories, scientists are involved in 17.6% of laser incidents,

second only to technicians (21.3%) – Doctors and Nurses include 9.2% of all incidents, 5th highest of all

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Lasers: Causes of the Most Common Accidents

1. Accidental eye exposure during beam alignment2. Misaligned optics3. Failure to wear available eye protection

It only takes 0.25 seconds (time required for the blink response) with a direct hit from a high powered laser

to damage your eyesight.

Don’t become a statistic, protect your eyes!

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