Transcript The Laboratory Environment

The Laboratory Environment
LAT Chapter 7
LAT Presentations Study Tips
Chapter 7
• If viewing this in PowerPoint, use the
icon to run
the show.
 Mac users go to “Slide Show > View Show” in menu bar
• Click on the Audio icon:
when it appears on the
left of the slide to hear the narration.
• From “File > Print” in the menu bar, choose “notes
pages”, “slides 3 per page” or “outline view” for taking
notes as you listen and watch the presentation.
 Start your own notebook with a 3 ring binder, for later study!
Humidity & Temperature
Chapter 7
• Standard relative humidity = 30 to 70 %
• Standard temperature range =19°C to 26°C
• Higher temperatures for post-operative recovery for birds,
reptiles, many nonhuman primates, and hairless rodents.
• Measurement:
• thermometer, minimum-maximum
thermometer, thermograph
• Relative humidity:
• amount of water present in air
• hygrometer, wet/dry bulb thermometer
•Computerized systems automatically
record environmental data.
Air Exchange
Chapter 7
• 10 to 15 fresh air room changes / hour standard
• Either 100% fresh air or re-circulated air
• Air filtered before and after leaving the room
• Reduce level of airborne pathogens, odors, chemical
contaminants, and particulates
• Supply and exhaust measured with anemometer
• measures the velocity of air passing through the vent
expressed as cubic feet per minute (CFM)
• Adjustments to supply and exhaust flows are what
determine positive or negative room pressure.
Dealing With Ammonia
Chapter 7
• A gaseous by-product of the bacterial
metabolism of urea, a substance found in urine.
• Heavier than air and becomes concentrated
• Can be a serious problem in filter-top cages.
• Mycoplasma pulmonis may cause disease in
the presence of a high ammonia.
• Some commercial bedding material contains an
ammonia inhibitor.
Laminar/Mass Air Displacement
Chapter 7
• Uniform, unidirectional, continuous flow of filtered air
• 200 or more air changes per hour
• Laminar air flow combined with HEPA filters prevent
airborne microorganisms from entering.
 HEPA filters are 99.7 percent efficient.
 removes particles as small as 0.3 microns.
 If HEPA filters are kept dry, bacteria and viruses cannot pass
through them.
• Laminar air flow cabinets or cubicles  Air is drawn through a pre-filter and forced into a plenum or
distribution chamber, then through HEPA filters and over cages.
 With reversal of air flow, devices achieve biohazard containment.
Laminar/Mass Air Displacement
Chapter 7
Ventilated Cage Racks
Chapter 7
• Provide HEPA-filtered air to each individual cage.
• Provide a barrier at the cage level.
 Continuous air flow reduces ammonia levels.
• Dust and hair can clog a HEPA filter.
 pre-HEPA filter for gross contaminants
• Laminar flow hoods (work stations) used for manipulations
 Most are of the type which protects both the operator and the hood
contents from contamination (Class II).
• Understand how the hood functions
 Stacking cages and other materials inside a hood can disrupt the
normal air flow => hood ineffective.
 Will not prevent exposure of personnel to chemical agents such as
formaldehyde or gas anesthetics.
Other Environmental Variables
Chapter 7
• Intense lighting => retinal degeneration in albino spp.
• Most animals tolerate lighting of 35- to 100-foot candles.
• 12-hour light/dark cycle - on 12 hrs and off 12 hrs.
 Automatic light timing devices prevent lighting variables.
 Variations => reduced breeding in some species.
• Animals can be sensitive to noises humans can’t hear.
 Some rodents susceptible to audiogenic seizures when exposed
to sudden loud noises.
 Noise stress => enlarged adrenal glands, reduced breeding
efficiency, increased blood pressure, auditory damage, and
behavioral disorders.
• Avoid sudden loud noises greater than 80 decibels (db).
Sanitation - Disinfect or Sanitize
Chapter 7
4 levels of sanitation:
1. Cleaning: complete removal of visible soil from surface
2. Sanitation: reducing organisms living on inanimate object
to an acceptable public health standard
3. Disinfection: reducing number of pathogenic organisms
(not necessarily spores) to a harmless level
4. Sterilization: rendering an object free of all living
organisms
Sanitation - Sanitize
Chapter 7
• The animal facility is continuously recontaminated by air,
water, animals, and people.
• Sanitation program = cleaning + sanitizing.
• Degree of risk = type and level of contamination + use.
• > risk of infection if organisms are resistant +/or highly
virulent.
• > risk if the animals are
particularly susceptible (e.g.
immunocompromised).
Choosing a Chemical
Chapter 7
Label Claims: regulated by EPA under the Federal Insecticide,
Fungicide, and Rodenticide Act
Spectrum of Activity: the specific organisms tested against the product
Effectiveness in Hard Water: Hard water ions can inactivate chemical.
Stability of the pH: Buffers prevent > pH changes from the concentrated
to the diluted form or by additives such as soaps.
Use Dilution: Using too much of product is wasteful and using too little
may reduce or eliminate the antimicrobial effect.
Contact Time: Essential that agent be in contact with surface long
enough to kill the most resistant organisms present.
Temperature: Heat could cause the evaporation of some of the
components of the formulation.
Other Attributes
Chapter 7
Toxicity: thoroughly rinse away
Application Mops and squeegees
Sprays
Immersion
Fogging
Fumigation
Evaluation Methods: bacterial cultures
Sterilization
Chapter 7
• Moist heat, dry heat, chemicals, and radiation
• Steam autoclave is primary means of sterilizing.
• Resistant organisms indicators for testing sterility.
• Most common biological indicator are spores of the
bacterium Bacillus stearothermophilus.
 Small vials placed into the autoclave during a sterilizing cycle.
 Vial incubated to detect any growth of the spores.
 A color change indicates growth of the bacteria.
 No growth = sterilizing cycle is operating properly.
• Chemical reaction temperature indicators - change color
when correct surface temperature reached.
Methods: Moist Heat
Chapter 7
• Hot water is effective only as a sanitizer.
• Steam is a good sterilizer.
• Steam under pressure - temperature > 100°C (212°F).
• Limitations
 cutting edges dull
 dry fabrics scorch
 some wet materials corrode
 some rubber and plastics deteriorate
 certain materials don’t mix with water
 possibility of serious injury
• Minimum sterilization time 15 minutes at 121°C (250°F)
or 5 minutes at 132.2°C (270°F).
Autoclaves
Chapter 7
1. Central chamber is surrounded by a jacket.
2. Steam saturated with water vapor and superheated
under pressure.
3. Steam baffle prevents load from being saturated.
4. Drain present at the lowest point of the chamber.
5. Valves top and bottom permit the exit of air and steam
6. Safety valve if the steam pressure exceeds a safe level.
7. Air inlet and vacuum air filters remove particulates/
8. In-line thermometer in the steam drainpipe
9. Door gaskets, joints and seals must be air-tight.
To Run an Autoclave
Chapter 7
• Steam in at top, displaces air out drain in bottom.
• Only effective if all the air is removed.
 Air pockets prevent steam penetration and heat transfer.
• Load equipment to be sterilized.
• To start, close door tightly and turn on timer.
• When the temperature reaches 121°C and pressure
reaches 15 pounds per square inch (psi), the timer
begins sterilization time.
• Steam is then vented.
• Drying cycle reduces residual moisture.
Autoclaves
Chapter 7
Air and
Steam In
Chart records the
run cycle, times and
temperature.
Safety tip:
Jacket stays hot!
Air and
Steam Out
Avoid Trapped Air
Doors can be
automatic or
manual.
Make sure the gasket and
drain are clear of debris.
Dry Heat
Chapter 7
• Kills most commonly encountered microbes.
• Requires long time (one to two hours at 160°C)
to effectively sterilize.
• Can scorch or burn certain materials.
• Most common dry heat method is hot air oven.
• Effective on equipment or bedding materials
damaged by moist heat or chemicals.
Chemicals
Chapter 7
• Glutaraldehyde
• Formaldehyde
 Toxic, carcinogenic, corrosive, and
has limited penetrability.
• It should be used only by
specially trained personnel.
• Peracetic acid
• Chlorine dioxide
• Ethylene oxide
• Plasma sterilizers
Radiation
Chapter 7
For interesting information on food irradiation, go
to: ccr.ucdavis.edu/ irr/what2.shtml
• Ionizing = gamma and beta
• Non-ionizing = ultraviolet, or UV
• Disrupts microorganisms protein structure
• Ionizing radiation can be lethal to humans
• Gamma rays used on instruments and supplies.
• Irradiated diets are nutritionally supplemented.
• Non-ionizing radiation less penetrability.
• Irradiated items are not radioactive.
Vermin Control
Chapter 7
• Carriers of disease-causing agents
• Walls and floors free of cracks and crevices
• Pipelines, drains, and air filters well sealed
• Inspect incoming supplies for vermin
• Keep stored feed, bedding, and caging away from walls
• Noninvasive = traps, sticky boards, boric acid, or silica
• Eliminating feral or wild rodents = poisoning or trapping in
areas outside the animal rooms.
• Place traps with triggers or entry ports along walls
• Control involving pesticides pest control personnel
Safety & Hygiene
Chapter 7
• Instruct on precautions taken in work area and use of
safety equipment.
• Advise that use of safety equipment is mandatory.
• Equipment must be available for any type of risk or
exposure encountered.
• Employee’s responsibility to perform in a safe manner.
• First aid stations / emergency eye-wash or shower stations
/ fire extinguishers / spill kits and instructions / emergency
evacuation routes
• Good personal hygiene needs to be enforced.
Research Environment Hazards
Chapter 7
Radioisotopes, living pathogens, carcinogens, and toxins
• Sign information required:
 identity of biohazardous agent, the name and telephone number of
responsible supervisor, and special requirements for entering
• Basic - microorganisms not known to cause disease in
healthy adult humans
 CDC classifies these organisms as BSL1 (Biosafety Level 1).
• Containment - separate environment from public
 These organisms are classified as BSL2 (Biosafety Level 2),
• High containment - may cause serious or fatal disease
 These organisms are classified as BSL3 (Biosafety Level 3).
Research Environment Hazards
Chapter 7
What is a radioisotope?
Link to web site on the various uses of radioisotopes.
http://www.uic.com.au/peac.htm
Isotopes are different forms of an atom of the same chemical element. They have
identical chemical properties but a different relative atomic mass. While the
number of protons is the same, the number of neutrons in the nucleus differs.
Some isotopes are referred to as 'stable' and others as 'unstable' or 'radioactive'.
It is the radioactive nature of these unstable isotopes, usually referred to a
'radioisotopes', which gives them so many applications in modern science and
technology.
see also ANSTO paper on Radioactivity, Radioisotopes etc
Primary Barriers
Chapter 7
Biological safety cabinets
 Class I,II & III cabinets
 Effective operation of safety cabinets depends on
inward flow of air, and any activity that disrupts that
flow can result in escape of material from the cabinet.
 Cabinets should be tested at installation, any time it
is moved to a new location, and at least annually.
 A certification label, with the date that the next routine
check is due, is attached to the cabinet at the time of
testing.
Secondary Barriers
Chapter 7
• Air locks, locker rooms, shower areas,
ultraviolet lighting, differential airflow, air filters,
and other such facilities outside immediate
animal housing environment
• Combination of structural barriers plus primary
barriers and good technical safety skills
routinely protect technicians.
Secondary Barriers
Chapter 7
Secondary Barriers
Chapter 7
Ultra Violet Irradiation
Waste Disposal
Chapter 7
1. Research protocol must specify type, amount, route and method of
excretion.
2. Isolate hazardous animals from animals that don‘t contain materials.
3. Post sign on room/cages containing hazardous materials.
4. Post “Notice To Employees” and “Emergency Procedures” signs.
5. Under supervision of veterinarian and Occupational Health and Safety.
6. Monitor personnel working with animals that emit gamma radiation.
7. Instruct personnel in proper procedures for handling and disposing of
contaminated wastes from the cages.
8. Occupational Health and Safety Office approval of investigators .
9. Occupational Health and Safety Officer central control over methods
and records of disposal of hazardous animal carcasses.
10. Instruct personnel in any additional precautionary measures.
Environmental Enrichment
Chapter 7
• Allows animals to engage in “species typical” behavior.
• The Guide divides behavioral management into:
1 Structural environment
2 Social environment
3 Activity
• Animal Welfare Act mandates written plan and records.
• Dogs - exercise schedules outside of cage and
technician interaction
• Primates - indestructible objects, food treats, puzzle
solving, group housing of compatible animals, swings,
perches and technician interaction
Additional Reading
Chapter 7
1. Biosafety in Microbiological and Biomedical
Laboratories, HHS publication No. (CDC) 938395, 1993.
2. Fox, J.G., Cohen, B.J., and Loew, F.W., eds.
Laboratory Animal Medicine. Academic Press,
Inc., Orlando, FL, 1984.
3. Occupational Health and Safety in the Care and
Use of Research Animals. National Academy
Press. 1997.