HSEoflabanimal25 10 97

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HSE of lab animal 25-10-97
Presentation · May 2020

DOI: 10.13140/RG.2.2.36393.01128
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Mohammad sadeq Khosravy

Institut Pasteur of iran
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Health, Safety, Environment
(HSE)
in Laboratory Animals
By M.S. Khosravy ( D.V.M)

mskhosravy@yahoo.com
Health, Safety, Environment (HSE)
In Laboratory Animals
Biosafety for animal experiments:

• What is biosafety for animal experiments?
The application of combinations of :
 Laboratory practice and procedure
 vivarium facilities,
 safety equipment
when working with potentially infectious microorganisms.
W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬


Hazards Associated with Laboratory Animals:

Physical Hazards:
• Ergonomic issues
• Strained postures
• Heavy lifts
• Repetitive motion
• Slips/Trips/Falls
• Noise/Heat in Cage Washing
Rooms
• Bites

Physical Hazards:
• Heavy lifting and awkward
postures are part of the daily work.
• More strains and awkward
postures. An administrative
control would be to prevent the
use of the top and bottom row for
storage, providing low stools with
wheels for handling the cages on
the bottom rows.

Biosafety Hazards when Working with

Laboratory Animals:
• Sharps: claws, teeth, surgical instruments,
edges on animal cages
• Zoonoses: human pathogens which are often
asymptomatic in the animal
• Surface Contact: blood, body fluids
• Allergens: dander, fur, dust, bedding
• Vectors: insects, parasites
General Safety Information for Rodent Handlers:

 Laboratory Animals Potential Hazards:
 Zoonoses for Zoonosis “Zoo” = animals “nosis” = disease
• Allergies (Allergic responses)
 Bites & Scratch & other trauma
 Chemical Agents
 Laboratory Animals facilities potential hazards
• General Safety :Personal hygiene, personal
protective Equipment [PPE]).
Routes of Exposure in Animal Procedures:

• Parenteral Inoculation (needle stick)
• Subcutaneous wound (bite or scratch)
• Surface Contact (contaminated work area)
• Ingestion (hand to mouth, food in lab)
• Inhalation (aerosol-generating procedures)
• Ocular (hand to eye, cleaning infected bedding)
• Mucous Membrane (aerosol droplets in face)
• Skin absorption

The Risk Assessment:

• Virulence/pathogenicity/infectious dose
• Environmental stability of the agent
• Route of spread, communicability
• Quantity/concentration/volume of agent used
• Vaccine/Treatment availability
• Allergenicity.

Risk Assessment:
• Routes of Transmission of the Agent
• Medical Surveillance
• Engineering Controls
• Personal Protective Equipment Use
• Facility Design, both laboratory and vivaria
• Zoonoses
• Vectors
• Characteristics of animals:
The species, strain, age or sex of an animal may affect the level of risk presented
during the procedure. For example, a pregnant or nursing animal may be more
aggressive towards handlers than one without young

Minimize Risk:
• Wear proper PPE (respirators, gloves, gown, shoe
covers, etc.)
• Use gloves when handling animals & change gloves
between animals.
• Wash your hands as soon as possible after removing
gloves. Hands must be washed with disinfectant hand soap prior to leaving
the room.
• No eating, drinking, smoking, applying cosmetics or
handling contact lenses in any laboratory animals
facility.
• Keep food intended for human consumption separate
from animal food.
• Report any animals that appear to be ill.
• Report any occupational illness or injury to your lab
manager.
Immunocompromised/Pregnant
Individuals:
• Immunocompromised individuals and
pregnant women should be aware of the
potential zoonotic hazards that may be
present in the workplace.
• If you are undergoing chemotherapy/
radiation therapy, being treated with
steroids or other drugs that could cause
immunosuppression and/or you are
pregnant or plan on becoming pregnant,
please determine appropriate protective measures
and monitoring.

Good Work Practice Limit Access to

Animal and Lab Facilities:
– Ensure that the PI and Animal care
personnel limit access to animal
biohazard containment areas:
– Access to animal areas shall be
limited to essential personnel only.
– Visitors may not know the rules of
safety in lab areas and may also
bring contamination from the outside
which may affect the health of the
animals.
Good Work Practice:

 Label All Animal Biohazard Areas
PI posts appropriate biohazard signs with
the universal biohazard symbol on the
entrance to biohazard use areas to warn
the public that biohazardous materials
are being used within the room.
All PI’s must:
 Label all contaminated equipment in
the room (incubator, freezer) with a
biohazard label.
 Dispose of pathological waste in a red
bag labeled with the biohazard symbol.

Signage:
• Signage is important to
communicate the potential hazard
• Post a biohazard warning sign
when immunizations or use of
respirators is needed for entry
• Provide contact information
• Persons at increased risk of
infection should not enter
infected animal rooms
Good Work Practice

Hand washing
• Ensure hand washing procedures are
followed
Hands should be washed:
• with warm water and a disinfectant hand
soap
• immediately and thoroughly if
contaminated with blood or other body
fluids.
• immediately after gloves are removed.
A foot and elbow operated sink is required for
Animal Biosafety Level 3 work.

Safety Equipment:
• Vivarium workers should wear
two pairs of gloves when
appropriate.

Safety Equipment:
•Protective clothing with a
solid
front may be worn in the
vivarium.
•Eye protection as needed
•Respiratory protection as
dictated
by the risk assessment
•Head covering as needed
•Clothing is changed and when
contaminated.

Facility Design and Use

• The animal area should be designed so
that it can be easily cleaned and
decontaminated.
• Bench tops must be impervious to
water and resistant to heat, organic
solvents, acids, alkalis, and other
chemicals.
• Chairs used at BSL 2 must be covered
with a non-porous material that can be
easily cleaned and decontaminated
with your approved disinfectant.

Good Work Practice

Decontaminate Work Areas
Verify that the disinfectant used will be effective for the
agent used. Many disinfectants do not kill a broad
spectrum of organisms. Animal laboratory work surfaces
should be decontaminated:
• after a spill of blood or biohazardous materials
• when work activities are complete
• with an appropriate chemical disinfectant
! If animals are left in the room, be sure that fumes from the disinfectant will not harm
the animals.

Decontamination of the Red Bag Waste:

Allergic Reactions to Rodents:

• The greatest occupational risk in working with rodents is the development allergies to
rodents.
• 15% of the general population is allergic to animals.
• 30 –50%( mean: 40% ) of those without a previous allergy history will develop an allergy
to lab animals while working in that environment.
• 70% of those with pre-existing allergies will develop a new allergy
• and about 10-15% of these workers may have symptoms of occupational asthma.
• The main groups at risk:
• Researchers – staff, students and visitors
• Animal technicians
• Maintenance staff
• Contractors
• Visitors

Risk Factors for Development of

Laboratory Animal Allergies:
Exposure to allergens
• Duration
• Frequency
• Intensity
Previous allergic conditions
• Personal history of allergies/atopy/eczema
Other predisposing conditions
• Illness
• Immunocompromised
• Pets
• Family history

Allergy Screening:
• Personal history of allergies/ atopy/eczema
strongly associated with increased risk
• Family history of allergy also important
• having the pets
• Only test in case of pre-existing symptoms
when patient plans on working with that
species
• Periodic screen by history for development of
new allergies in high risk patients
Allergy Symptoms:
Symptoms can be provoked by inhalation of allergen or by allergen
introduced into a break in the skin caused by scratches, bites or
instruments.
– Red, itchy, watery eyes, runny nose1
– Sneezing, itchy, runny nose, congestion1
– Red itching skin, welts, hives2
– Asthma3”cough (can be late-phase with symptoms
starting several hours after leaving the animal
facility), wheezing, chest tightness, shortness of
breath
– Anaphylaxis4” itching, hives, throat tightness,
fainting, nausea, vomiting, diarrhea
1 = common, 2 = somewhat common, 3 = about 15 –30%, 4 = rare

• These allergens can be carried by:

– air currents in the animal room
– and can come into contact with your skin, eyes, nasal passages, and lungs
– where allergic reactions can occur or when you are exposed to rodents or to
rooms and equipment used to house the rodents
• People with allergies to rodents may have symptoms:

o Skin rash/itching or hives
o Watery or itchy or red eyes
o Runny nose or sinus congestion
o sneezing
o Wheezing or chest tightness
o Shortness of breath or difficulty breathing

Common Lab Animal Allergy Sources

• Rats/ Mice: major allergens in urine/saliva
• Cats: sebaceous glands, hair, saliva
• Dogs: saliva, hair, skin
• Rabbits: fur, saliva, urine
• Birds: droppings
• Bedding: (mattress dust and
mite derived allergens)

• People who have pre-existing allergies are at greater risk

when handling rodents.
 Rodent allergens :
• (protein materials which cause an allergic reaction in
people) are found in the animal’s:
 Urine
 Fur
 Saliva
 Hair
 Dander
 Droppings
 Serum.
 pelt proteins can also be allergenic


These areas of high-risk work for allergy are:

• Cleaning of animal rooms:
o Cleaning animal rooms by dry sweeping floors with a brush is unacceptable as it
generates a high airborne allergen load close to the worker.
o Floors should be cleaned by damp sweeping or HEPA filtered vacuum cleaning.
• Cleaning of animal cages:

 Cage cleaning: under a purpose-built extraction hood
 Mobile cage cleaning stations: filtered the exhaust air through HEPA filters
 Vacuum cleaning: Care must be taken with these units that noise levels to
the animal rooms( by HEPA filters and a two-stage bin arrangement)
• Disposal of waste:
o Soiled bedding from cleaning operations should be handled with due precautions and
carefully sealed in plastic bags (preferably double bagged) for disposal.

Prevention of Lab Animal Associated Allergies:

– Biosafety cabinets
– Filter top cages
– Ventilated cage racks
– Biobubble
– Choice of bedding
– Reduce time with animals
– Wash hands frequently
– Animal density
– Proper housekeeping practices Cage dumping in open waste bins can generate a
– Proper humidity significant number of airborne particles.
– Personal protective equipment (masks, respirator)

How Can You Prevent Allergies?

• Limit your exposure to allergens.
• • Enforce controlled access to animal facilities: Biosafety Cabinets &
Respirators
• Use good personal hygiene:
o Always wash your hands after contact with animals and before leaving the
laboratory or animal facility.
o Do not eat, drink, smoke, handle contact lenses, or apply cosmetics in work
areas, and wash your hands before engaging in any of these activities.
o Avoid wearing street clothes while in an animal facility, especially if working
with animals.
• Follow appropriate housekeeping procedures:
 After working with animals, do not dry-sweep working surfaces. Instead, use
a damp cloth or another wet method for cleaning and sanitation.
• Perform animal manipulations in well-ventilated areas and, if possible, in
ventilated hoods or safety cabinets.

How Can You Prevent Allergies?

• Wear personal protective equipment:
o Wear lab coats, scrubs, or coveralls when working with animals, and
have work clothes laundered in the workplace.
o Wear goggles and a mask, especially if splashing may occur.
o Wear disposable gloves when in contact with animals or their excreta.
o Wear a respirator while in areas where the concentration of airborne
allergens is elevated (e.g., highly populated animal rooms, cage
dumping areas, cage washer).
o Remember that disposable surgical masks may not offer protection
against allergens.
o Properly fitted masks, such as N95 particulate respirators, provide
superior protection.
o Participation in fit testing.


Treatment of Lab Animal Associated Allergies
• Prevention is preferred
• Education of employees
• Proper use of personal protective equipment
• Re-assign employees when needed
• Medical treatment to reduce symptoms

General description of zoonotic diseases:

 Of the >1400 pathogens known to affect humans, >60%
are zoonotic
 Zoonotic diseases are caused by infectious agents such
as:
– Virus
– Bacteria
– Parasites
– Fungi
– Prions (“infectious protein”)
– Mammals, birds, reptiles and fish may all carry zoonotic
diseases (as well as insect vectors)


Modes of transmission of zoonotic infections :

1. Direct transmission
 Direct contact:(including faeces, saliva etc)
– Influenza
– Tularemia
– Salmonella
Aerosoles:
– Influenza virus
– Hantavirus (musepest)
– Tularemia
Bites:
– Rabies
Staphylococcus aureus
– Tetanus
– Streptococcus

Bacterial Infections from Bite Wounds

>200 species of bacteria in the mouths of many animals,
including humans.
– Streptococcal species, staphylococcal species, tetanus.
– Bite wounds should be thoroughly cleaned and debrided.
– Cat scratch disease--Bartonella henselae
– Rat Bite Fever-Spirillum minor, Streptobaccilis moniliformis
– Prophylaxis for moderate to deep bites with
Amoxacillin/clavulinic acid (Augmentin).

Animals Bite & scratch:

• Yersinia pestis--fleas from street cats/wild
rodents

Modes of transmission of zoonotic infections continued

. Indirect transmission
 Food- and waterborne:
– Salmonella
– Camphylobacteria
– Tularaemia
– Helicobacter. muridarum salmonellosis
 Vector borne:
– Mechanical vectors:
– Biological vectors (like ticks, flies, mosquitoes)
– Borreliose (Lyme disease)
– Tularaemia or Rabbit fever ،(Francisella tularensis )
– Yersinia pestis(fleas from street cats/wild rodents) Helicobacter

Risk of zoonotic infections in research:

 Laboratory work
• Handling of laboratory animals
• Working with animal
tissue/secretions/excretions
• Cell culture and microbiology (animal tissue,
cell cultures, microbiological cultures etc)
 Field work
• Handling of wild animals and wild animal
samples (tissue samples, organs, blood,
body fluids/excretion, faeces, etc.).

• Zoonosis risk when working with

live animals in the laboratory
continued
• Some animals used do not have
documented health status and may carry
zoonotic pathogens
• Even animals with documented
health status may carry or acquire mite
zoonoses, and/or have an

microbiologic flora that contains
potential zoonotic agents

‫)‪Health, Safety, Environment (HSE‬‬
‫‪In Laboratory Animals‬‬
‫‪ ‬پرورش و تکثیر منطبق با استانداردهای ملی ایران‪:‬‬
‫حیوانات آزمایشگاهی تولید شده در ایران از حیث درجه کیفی «آلودگی های میکروبی» جسء گروه‬
‫قراردادی ها( ‪ )conventional‬و یا متعارف ها قرار می گیرند‪.‬‬
‫‪ : Conventional Animals ‬طبق تعریف سازمان دامپسشکی ایران حیًاواتی َستىد کٍ‬
‫فاقد عًامل بیماری زای قابل سرایت بٍ اوسان می باشىد‪.‬‬
‫‪ ‬از انجام اقدامات درمانی در حیواناتی که به زئونوزها مبتال شده اند! باید جلوگیری شود‪...‬‬
‫‪ ‬کمتریه کیفیت ممکه ( با درجٍ آلًدگی یک ستارٌ) در استاودارد سازمان دامپسشکی ایران‬
‫برای حیًاوات قراردادی عبارت است از عدم يجًد آلًدگی َای ذیل‪:‬‬
‫•‬ ‫)‪Salmonellosis (Salmonella spp‬‬ ‫•‬ ‫‪Lymphocytic choriomeningitis virus‬‬
‫•‬ ‫)‪Tuberculosis (mycobacterium spp‬‬ ‫•‬ ‫‪Dermal fungal disease‬‬
‫•‬ ‫)‪Pasteurellosis (Pasteurella Yersinia‬‬ ‫•‬ ‫) ‪Leptospirosis ( leptospira spp‬‬
‫•‬ ‫‪(listeriosis) Listeria monocytogenes‬‬
‫•‬ ‫‪Streptococcus moniliformis‬‬
‫انستٌتو پاستور اٌران‪ -‬بخش تحقٌقات و مرکز رفرانس هاری ‪W.H.O‬‬ ‫‪44‬‬
‫‪ ‬پرورش و تکثیر خوکچه هندی مبتنی بر درجات آلودگی میکروبی منطبق با استانداردهای ملی ایران‪:‬‬
‫‪ ‬پرورش و تکثیر خوکچه هندی مبتنی بر درجات آلودگی میکروبی منطبق با استانداردهای ملی ایران‪:‬‬
laboratory animal health monitoring programme :

laboratory animal health monitoring programme :

Zoonotic Diseases of Rodents:

• Mice, rats, hamsters, guinea pigs and gerbils can
carry organisms that may cause infection and
disease in humans (zoonotic diseases, zoonoses)
and these may be transmitted either directly
(e.g., through bites or scratch) or indirectly (e.g.,
through exposure to feces).
• Natural infections & Most infections in Rodents
are usually subclinical.
• Various host and environmental stressors may
precipitate clinical disease. Rounded (Abnormal) Position
• Clinical signs occur most commonly in suckling
and weanling rodents and include sudden death,
watery diarrhea, lethargy, and ruffled fur.
Staphylococcus aureus

Occupational Health: Zoonoses

• Mice can be a reservoir of the following
infectious agents that are transmissable to
people:
– Viruses
Hantavirus
Lymphocytic choriomeningitis virus (LCM)
– Bacteria
Leptospira spp
Salmonella spp
– Fungi
Microsporum spp, Trichophyton spp
(LCM)
– Parasites
Rodentolepis nana
– Quarantine can last as little as 24 hours but is
usually for a week

• The type of animals involved and the nature of

the procedure will affect the risk of exposure to
infectious hazards. Hydrocephalus
• For example, an experiment with Salmonella will
generate much larger quantities of infectious
bedding using a calf compared with using a rat.
• The nature of each procedure will also present A hunchbacked mouse with piloerection
different risks.
• Animal husbandry, bleeding procedures and post
mortem examinations all present opportunities
for exposure but in different ways and so may
require different methods of control.
Runting in infected by MHV naïve, juvenile mice

Zoonoses in natural populations of animal species most commonly used for
experimental procedures.
A dehydrated mouse
W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬ 52

Use of animals from approved vendors and proper use of personal protective equipment
makes the risk of transmission of these diseases unlikely.
PATHOGEN TRANSMISSION ANIMAL DISEASE HUMAN DISEASE
Streptobacillus moniliformis, • animal bites(Rat bite fever) usually a subclinical infection, but polyarthritis, myalgias(Muscle Pain)
Spirillum minor • ingestion of contaminated purulent lesions have been reported in regional lymphadenopathy, fever
(Rat bite fever, Haverhill fever) food products(haverhill fever) some animals
Salmonellosis fecal-oral, ingestion of Malaise(a general feeling of discomfort, dehydration, vomiting, abdominal pain,
(most rodents) contaminated products illness), dehydration, bloody diarrhea nausea
Leptospirosis direct contact with contaminated infertility, fever, anorexia, anemia headache, myalgia, conjunctivitis,
(most rodents) urine nausea
Lymphocytic Choriomeningitis exposure to saliva or urine from viremia, viuria, and chronic wasting subclinical infection, mild flu-like
(LCM) infected animals or to infected disease symptoms
cell lines in the lab fomites may viral meningitis and encephalitis (rare)
play a role
Hantavirus exposure to aerosols, urine, and subclinical fever, myalgia, petechiation, abdominal
(most rodents) fecal material from infected pain, headache
animals
fomites may play a role
Dermatophytosis (Trichophyton direct contact circular raised erythematous lesion with circular raised erythematous lesions
mentagrophytes) hyperkeratosis and hair loss with hyperkeratosis and hair loss
Ornythonysisus Bacoti direct contact with cage materials assymptomatic to moderate pruritis severe pruritis
(Tropical Rat Mite)
Sarcoptes scabei (Guinea Pigs and direct contact with infected intense pruritus intense pruritus
Hamsters) animal

Zoonosis in the laboratory continued

• Infectious experiments - dedicated
research on zoonotic agents (in
animals or cell culture) Infectious
disease research - Biosafety level 2,
3, 4
• 3.“Regularly” biology (animals,

organs, tissues, etc.) and general
cell biology work risk of zoonotic
contaminated material

Exposure Procedures Immediate Response:

Mucous membrane:
 flush in an eye wash or potable water for a
minimum of 15 minutes.
Non-intact skin exposures:
• Wash with soap and water or antiseptic for 15
minutes.
• REPORT ANY INJURIES AND ILLNESSES TO
PI/LAB SUPERVISOR
Bacterial Infections from Bite Wounds:

>200 species of bacteria in the mouths of
many animals, including humans.
Streptococcal species, staphylococcal species,
tetanus.
– Bite wounds should be thoroughly cleaned.
– Prophylaxis for moderate to deep bites with
Amoxicillin /clavulinic acid (Augmentin).

Tetanus
• Acute disease caused by a toxin produced by
the bacteria Clostridium tetani
• Potentially in the intestines of most warm blooded animals, in
particular horses, sheep, cattle, dogs, cats, rodents, guinea pigs,
and chickens
• Typical infection occurs after wounds are contaminated with
faeces, soil, dirty needles/scalpels, or after bites often involves a
cut or deep puncture wound
• All animals, including humans, may become sick and tetanus
affects the skeletal muscles causing spasms and stiffness. May be
lethal.

Tetanus prevention:
– Vaccination: every 10 years, and additional immunisation
(“boosting”) after accidents depending on degree of
damage and vaccination status, see www.folkehelsa for
details
– Careful handling of live animals and excretions, biological
samples, etc.
– Good hygiene, including the use of gloves and hand wash
– Thorough and immediate cleaning and disinfection of
wounds
– Report any accident and seek medical aid for evaluation


Tularemia («hare plague»)

• Caused by the intracellular bacteria
Francisella tularensis
• Hares are in particular sensitive and in this
species the disease is usually lethal, but
healthy carriers exist(during incubation
period)
• Other species may also carry the infection:
lemmings, wild mice (voles), beaver, etc.
• Transmission: direct contact with infected,
sick or diseased animals
• aerosols (e.g. old nests, dust, etc).
• contaminated food or water (“rodent years”)
Tularemia in humans:
Very few bacteria needed to give infection

•The bacteria can penetrate intact skin
•Direct contact transmission gives skin infection, with
accompanying swollen lymph nodes, fever, headache
and nausea
•Aerosols may give pneumonia
•Water- and food borne usually as diarrhoea and
nausea

Salmonellosis:
• Caused by different variants of Salmonella spp bacteria
• S. Typhimurium one of the most prevalent in the world
• Food and water borne transmission most common, but
the disease can also be transmitted through contact with
infected animals (incl. humans), biological samples,
faeces, etc. (faecal-oral infection route)
• Human symptoms include diarrhoea, nausea, headache
and fever

• Hantavirus – Nephropathia epidemica “Mouse plague”

• Reservoir: Wild Rodents
• Disease transmission:
– Aerosols (particularly urine and faeces).
– Contaminated drinking water
– Autumn/early winter
– People at risk: Wildlife biologists
 Symptoms in humans :
– Influenza-like symptoms followed by acute kidney disease
– pulmonary syndrome

Injuries from Handling Rodents:

• Rodents may occasionally inflict injuries to people
such as bites or scratches.
• The long incisor teeth of rodents can inflict deep bites.
• It is essential that people who handle rodents for
research or teaching be provided with training in
proper handling techniques
• in order to avoid injury to themselves or the animals,
such specific handling and restraint techniques, and
protective clothing requirements

Characteristics of animals
• The species, strain, age or sex of an animal
may affect the level of risk presented during
the procedure. For example, a pregnant or
nursing animal may be more aggressive
towards handlers than one without young.





Good Work Practice

Minimize Inhalation Risks:
• Review inhalation risks to animal handlers when
personnel are:
– using high pressure hoses to clean surfaces
– cleaning animal bedding and cages
– handling the animals themselves
In addition, animals shed fur and hair which may
exacerbate allergies. For this reason, animal care
personnel should wear respiratory protection.
Cage Dumping
– The dumping of animal cages has
the potential to produce aerosols.
– Cages should be dumped in front
of the Biobubble at all times to
capture aerosols.
– The waste bin should be within 5
inches of the pre-filter of the
Biobubble.

Biobubble Maintenance
– The ventilation balloon in the back of
the Biobubble must be fully inflated at
all times when dumping cages.
– •A shrunken balloon indicates
restricted airflow through the pre-
filter.
– •The pre-filter must be cleaned or
replaced when the balloon indicates
restricted airflow.
– •The Biobubble must be certified
annually.

Cage Changing Stations

• The animal cage changing station provides
vertical HEPA filtered airflow for product
protection on the work surface.
• The station provides allergen personnel
protection by using a HEPA filtered down
draft vacuum on the work surface to
assure minimal exposure to animal
hair/dander and cage bedding material.
• •The cage changing station should not be
used as a substitute for a Class II, Type A2
(Non-ducted) Biological Safety Cabinet.
• •It is not a containment device for
biologicals

• Cage change stations:

• Cage change stations( video)

CAGE & RACK WASHER

Cage Washing

Proper Disposal of Animal Carcasses:

• The PI must first place the carcass in a red
bag avoid disposal in the regular trash.
• However, final disposal will depend of the
carcass was also contaminated with hazardous
chemicals or radioactive materials.
• ANIMAL CARCASSES MUST NOT DISPOSED
OF IN THE REGULAR TRASH!

Good Work Practice

• Minimize Sharps Use
Review all sharps usage:
• Sharps must be handled with extreme caution.
• Unless animals are restrained, they may squirm
and result in accidental self-inoculation (gas
anesthesia).
• Needles must not be recapped, bent, sheared,
or removed from a disposable syringe.
• All used sharps must be placed in a rigid,
hardplastic,puncture-resistant container for
disposal.

Animal Biosafety Level 1 (ABSL-1):

– Animal Biosafety Level 1 (ABSL-1) is suitable for
work involving little or no known potential hazard
to animal handling personnel and the environment.
– Work is conducted in the open (bench or cage).
– Special containment equipment,such as a
biosafety cabinet, is generally not required.
– E. coli K-12 is an example of a organism typically
used at ABSL-1.

Animal Room
• Lab coats or gowns are worn in the
animal facility.
• Gloves, Lab coat, eye protection
• Bedding materials are removed in such
a manner to minimize aerosol
generation.
• Hand-washing sink,safety shower/
eyewash, autoclave
• Cages are cleaned and disinfected on a
regular basis.

– Facility is separated from main
traffic Restricted access
– Doors – self closing and self locking,
kept closed, open inward Hand wash
sink
– No air recirculation
– Recommended negative pressure

Animal Biosafety Level 2

• Animal Biosafety Level 2 (ABSL-2) is similar to ABSL-1,
but work at this level requires containment devices
such as biosafety cabinets whenever aerosol generating
procedures are conducted.
• Specialized training required.
• Access to the facility is limited while the experiment is in
progress
• An example of an organism requiring ABSL-2 is Salmonella



Animal Biosafety Level 2:

• A biohazard sign is posted on the
outside door.
• All personnel must receive
appropriate training on the hazards
associated with the research.
• All biohazardous agents used in the
animal facility must be transported
in a manner to minimize leakage.
• Agents must be transported using
double containment in a labeled
container.


• Medical surveillance appropriate to the animal
species used shall be offered to workers prior to
the initiation of the experiment.
• Protective clothing worn in the animal use areas
shall not be worn outside the room.
• Work areas are disinfected at the conclusion of
the experiment.
• Autoclave nearby for trash, bedding and cages
Appropriate footwear


Facility Considerations
Check for the following safety equipment:
• A sink should be available for hand-washing.
• An autoclave should be available to treat infected
animal bedding prior to disposal.
• A biosafety cabinet should be available for aerosol-
generating procedures, such as vortexing animal
specimens.

• Consider room traffic flows
• Single entry vs. exit
• Clean/dirty double door
concept
• Clothes change
• Autoclave nearby



• Animal Biosafety Level 3 (ABSL-3) differs from
ABSL-2 in that the agents may cause serious or
lethal disease in animals or man.
• All ABSL-3 procedures are conducted within a
biosafety cabinet and the workers may also be
required to wear personal protective equipment
such as a respirator.
• The lab has special engineering features to
prevent a release of the agent to the
environment.
• An example of an agent requiring ABSL-3
containment is Mycobacterium tuberculosis.

Animal Caging Systems:

• When needed, partial or
full containment caging
systems as per risk
assessment
• Eg: Rodent-filtered top
caging, HEPA-filtered
primary barrier systems
appropriate to species

Ventilated Caging Systems


• All wastes from the ABSL-3 animal room are
autoclaved before disposal.
• All pathological waste from the animal room
shall be transported in a leakproof container
before disposal and eventual incineration.
• Appropriate face and respiratory protection is
worn by personnel entering the animal housing
areas.
• All experimental procedures shall be performed
in such a manner so as to minimize aerosol
generation.
• NO sharps.

• Animal Biosafety Level 4 (ABSL-4) is required
for all work with dangerous and exotic agents
which pose a high individual risk of infection to
animals and man which can be life threatening.
• Animal Biosafety Level 3 (Ag) is like ABSL-4 in
facility design and containment, but does not
necessarily protect the worker


• The following are examples of agents used at
– Ebola
– Marburg
– Lassa Fever Virus
– Machupo
Only a very few facilities may accommodate ABSL-4 experiments.

Medical Surveillance Considerations Vaccinations:

• The PI should be aware of the availability of
vaccines for certain organisms.
• OSHA requires that Hepatitis B virus (HBV)
• vaccinations must be offered to at risk personnel.
• Other vaccinations, such as rabies tetanus and
yellow fever are licensed and clearly have benefit
for the user.
• Some vaccinations, however, are experimental and
are may not be available for general use.

Emergency Procedures:
• The IBC and IACUC must review emergency procedures
for personnel who are bitten or scratched while
handling infected animals.
• Animal handlers must report the incident to their
supervisor and then to a qualified physician.
• The physician preferably would be familiar with
animal zoonoses and other health complications
associated with the handling of animals.
• The animal worker may have to provide a specimen in
conjunction with post exposure follow up, such as with
Herpes B Virus surveillance.
• Disinfectant Types:
– A disinfectant described with the suffix “cidal” is one that has
killing action.
– A bactericidal disinfectant kills bacteria but not necessarily
spores.
– A sporicidal disinfectant kills spores and bacteria.
– A disinfectant that has the suffix “static” merely inhibits
microorganism growth.
– A bacteriostat, for example, prevents the growth of bacteria
but does not necessarily kill them.
– Chemicals, such as phenols, sodium hypochlorite (bleach) and
quaternary ammonias, are commonly used to disinfect
nonliving objects such as floors and equipment.


Decontaminate Work Surfaces

Work surfaces must be
decontaminated:
• •After work,
• •After spills,
• •At the end of each day,
• •Prior to the repair or
surplus of equipment

Disinfecting Liquid Microbiological Waste

• Liquid waste such as human blood, animal blood, human tissue
culture, body fluids, or growth media that has not been used for
propagating microbes/vectors/toxins must notbe poured down the
sanitary sewer without prior steam sterilization or chemical
disinfection.
• For disposal of liquid microbiological waste that was used for
propagating microbes/viral vectors/toxins,NC Medical Waste Rules
do not allow chemical disinfection followed by disposal to the
sanitary sewer unlessapproval has been obtained from the NC
Division of Waste Management.
• The liquid chemical disinfection procedure used prior to disposal
down the sanitary sewer must be detailed on the Biohazard
Registration for the PI listed on their Lab Safety Plan.

Cage Washing
• Final rinse temperature for animal cages
should reach at least 180oF. This should be
documented in the cage wash notebook.

• Chemical and physical hazards:

Phenolic Compounds:
 the cats and certain other laboratory animals react
adversely to phenol.
‫ معموالً پاٌدار‬0 ‫ شماره‬:)‫ واكنش پذٌري (مربع رنگ زرد‬
‫ در صورت تماس با بخار يا مايع بسيار خطرناك‬،‫ خطرناك‬-4 ‫ شماره‬:) ً‫ سالمتً (مربع رنگ آب‬


Phenolic Compounds: ‫مواد سمى‬


Phenolic Compounds: ‫مواد خورنده‬


Phenolic Compounds:


Sodium hypochlorite:
 Bleach must be used carefully, as it can harm delicate tissues
such as the eyes and lungs.
‫ معموالً پاٌدار‬0 ‫ شماره‬:)‫ واكنش پذٌري (مربع رنگ زرد‬
‫از دستگاه تنفسي استفاده شود‬،‫ خطرناك‬-2 ‫ شماره‬:) ً‫ سالمتً (مربع رنگ آب‬
‫عامل اكسٌد‬

Sodium hypochlorite:) ‫( آب ژاول‬


Sodium hypochlorite:) ‫( آب ژاول‬ ‫مواد محرك‬

Sodium hypochlorite:) ‫( آب ژاول‬
‫مواد خورنده‬

Sterilization: Autoclaving
– Because steam under high pressure is used to achieve the temperature
needed for sterilization, laboratory animal facility personnel are advised to
handle autoclaves with great caution.
– Animal technicians should learn all about the particular machines they are
going to use before attempting to operate them.
– Minimum protective apparel for operating the autoclave is a lab coat, safety
glasses, gloves, and closed-toed shoes.
Testing Autoclaves
– uses different tests to ensure their autoclaves are working properly.
– Sterilization Monitoring should be used in each autoclave load. This will let the
technician know whether or not steam has penetrated into the items to be sterilized.
– Carefully place the indicator inside a cage, near the middle of the biohazard waste load.
– For animal areas generating biohazard waste at BSL 2 or above, a weekly autoclave
testing log must be completed every week and be available at all times.
– Autoclave testing logs must be kept for 3 years.



Handling and Picking Up a Mice

Handling and Picking Up a Mice


Device Restraint (FALCON TUBE)

Picking Up & Restraining A Mice

Picking Up & Restraining A Mice :Cylinder restrainer

Single-handed restraint of the mouse

Single-handed Restraint Of The Mouse

Single-
handed
restraint of
the mouse
Restraining flat-shaped cylinder for manipulating a Animal positioning for sexing
mouse
mouse positioned inside a body triangular plastic sleeve Fixation of a mouse during temperature
with hindleg exposed for possible blood sampling at the recording. Note: this is an adult animal
vena saphena

DecapiCone

Manual restraint of a mouse using both hands

Device Restraint (Cylinder restrainer or DecapiCone)
Restraining flat-shaped cylinder for Using a restraining wall for, e.g., blood
manipulating a BALB/c mouse sampling on the lateral tail vein of a mouse
mouse positioned inside a triangular Restraining conical cylinder for

plastic sleeve manipulations on the tail of a mouse

Restraint &
fixering of
the mouse

Oral Feeding ( gavage) in Mouse



Intraperitoneal administration (i.p) Injection

Intraperitoneal Injection ( i. p)

Intraperitoneal Injection ( i. p)

Subcutaneous administration (S.C) Injection

Subcutaneous administration (S.C) Injection

Intravenous administration (i.v.) Injection

Intravenous administration (i.v.) Injection

Intracerebral administration (i.c.) Injection

Intracisternal injection:By Stereotaxic alignment system
Animals must be heavily sedated or anesthetized for cannula or catheter placement and for direct injections.
Noninvasive-intratracheal

Aseptic Technique in Rodent Surgery



Tools for Blood Collection from Tail: Tail tip –mice

Blood Collection From Retro-orbital Sinus

Surgicalprep for Intracardiac (IC) Puncture Blood Sampling

Intracardiac (IC) Puncture Blood Sampling

Barbering and Shaving for Blood Collection From Saphenous Vein

Tapping for Blood Collection From Saphenous Vein

Tapping for Blood Collection From Saphenous Vein

Axillary Cut Down Blood Sampling Video

Blood Collection from Carotid artery
- This method is a terminal

procedure
- Where an incision is made
through skin and tissues in
the neck area.(a 5 cm incision
back Subcutaneously behind the
Ear and through the Incision
between the Scapulae.)
- The animal must be
euthanized after blood has
been collected and before it
regains consciousness.

Euthanasia By Carbon Dioxide



Handling and Picking Up a Rat













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HSE of lab animal 25-10-97

Presentation · May 2020

Mohammad sadeq Khosravy

Nanomedicine View project

LAB ANIMAL View project

The user has requested enhancement of the downloaded file.

By M.S. Khosravy ( D.V.M)

Biosafety for animal experiments:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Hazards Associated with Laboratory Animals:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Biosafety Hazards when Working with

General Safety Information for Rodent Handlers:

Routes of Exposure in Animal Procedures:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

The Risk Assessment:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Good Work Practice Limit Access to

Good Work Practice:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Good Work Practice

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Facility Design and Use

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Good Work Practice

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Decontamination of the Red Bag Waste:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Allergic Reactions to Rodents:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Risk Factors for Development of

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

• These allergens can be carried by:

• People with allergies to rodents may have symptoms:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Common Lab Animal Allergy Sources

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

• People who have pre-existing allergies are at greater risk

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

These areas of high-risk work for allergy are:

• Cleaning of animal cages:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Prevention of Lab Animal Associated Allergies:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

How Can You Prevent Allergies?

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

How Can You Prevent Allergies?

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

Treatment of Lab Animal Associated Allergies

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬

General description of zoonotic diseases:

W.H.O ‫ بخش تحقٌقات و مرکز رفرانس هاری‬-‫انستٌتو پاستور اٌران‬