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what are the two most commonly used species in biomedical research?
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rats and mice make up what percentage of all research subjects?
approximately 90%
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what are the four characteristics that make rats and mice so feasible?
- 1. small size
- 2. high reproductive capability
- 3. ease of handling
- 4. availability of special "breeds" called strains
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what is the rat genus, species?
Rattus norvegicus
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what is the mouse genus, species?
Mus musculus
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what is Mus musculus?
house mouse of north america and europe
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what is the most frequently used animal in biomedical research (genus, species)?
Mus musculus
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what is Rattus norvegicus?
the norway rat
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what was the first mammal to be used in research (genus species)?
Rattus norvegicus
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what are some specific areas that mice and rats are used in research?
- nutrition
- embryology
- toxicology
- oncology
- genetics
- organ transplant
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what should you do if you are allergic to rodents?
- wear PPE - mask, gloves and gown
- discard PPE when finished
- wash hands
- leave room if necessary
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what should you do if you do not know you are allergic to rodents?
- wear gloves
- wash hands
- outerwear worn only inside a rodent room (or gown)
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when is the tail hold used?
short term handling
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how do you use the tail hold?
- grasp tail at the base (closest part to the body, fat part)
- put rodent in new cage or place on surface for gripping
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what could happen if you grasp the tail at the tip?
skin can "slough" off
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which species do you use the scruff hold?
mouse
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how do you use the scruff hold?
- start with tail hold, placed on a surface for gripping (continue to hold tail)
- grasp the loose skin at the back of the neck
- grasp skin close to head to prevent bites
- hold tail between 4th and 5th fingers to steady mouse
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which species do you use the body hold?
rat
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how do you use the body hold?
- place hand over rat's body
- restrain head with 1st and 2nd fingers
- place thumb and 3rd finger behind front legs
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why would you use the scruff or body hold?
better/greater restraint
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what are three restraint devices?
- plexiglass restrainer
- decapicone
- centrifuge tubes
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what is the purpose of the plexiglass restrainer?
allows access to tail and other areas for injections or drawing blood
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what is the decapicone?
plastic "baggie"
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what is the purpose of the decapicone?
- restrains the rodent and leaves the tail free for injections
- IP/IM injections can be done through the baggie
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what is the purpose of the centrifuge tube?
keeps the rodent secure and useful for blood draw from saphenous vein in mice (be sure to cut hole in the end for air)
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what are the pros for using chemical restraint?
- no movement
- good access anywhere on body
- short or long periods
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what are the cons for using chemical restraint?
- must monitor anesthetic depth
- vaporizers are expensive
- chemical anesthetics need DEA number
- anesthetics may cause vasoconstriction
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what are the two main techniques for animal identification?
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what is included on the cage ID card?
- investigator's name
- species
- strain
- vendor
- age
- sex
- arrival date
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what are examples of individual IDs?
- ear tags
- ear punches
- tail tattoos
- toe amputation (hairless neonates only)
- nontoxic dyes
- shave patterns into fur
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what are the two ways to determine sex?
- anogential distance
- nipples
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which sex has nipples?
females
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how do you determine males using anogenital distance?
greater distance between anus and genital papilla
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why can you not rely on the presence of testicles to determine male sex?
testicles can be retracted into the body
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what is necessary for oral gavage?
- conscious, restrained animal best
- ball tipped feeding needle on end of syringe
- restrain and measure distance from mouth to last rib with feeding needle
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what size oral gave is used on mice?
20 ga, 1 1/2 in.
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what size oral gavage is used on rats?
18 ga, 3 in.
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how do you perform oral gavage?
- after measuring needle, place gavage needle in mouth and pass down esophagus by sliding it along roof of mouth
- animal should swallow as needle passes
- at correct depth, give fluid
- monitor for signs of discomfort, difficulty breathing, abnormal behavior
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how can oral medications be given to rodents?
- some antibiotics or pain meds can be placed in water bottle
- sweet flavored antibiotic can be given by dropper into the back of the mouth
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what are the four routes of injections in rodents?
- SQ - subcutaneous
- IP - intraperitoneal
- IV - tail vein
- Im - intramuscular
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what size needle is used on mice for SQ injections?
25-37 ga, 1/2 - 3/4 in.
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what size needle is used on rats for SQ injections?
22-25 ga, 1/2 - 3/4 in.
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how do you administer SQ injections on rodents?
- restrain rodent
- insert needle under skin of scruff and advance
- aspirate to ensure not in blood vessel
- inject fluids
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how much fluids can be given to mice by SQ?
2-3 mL
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how much fluids can be given to rats by SQ?
10 mL
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what size needle is used on mice for IP injections?
25-27 ga, 1/2 - 3/4 in.
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what size needle is used on rats for IP injections?
22-25 ga, 1/2 - 1 in.
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how do you restrain a rodent for an IP injection?
restrain rodent's body with head lower than body
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why should the head be lower than the body for an IP injection?
allows abdominal organs to move away from injection site
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where on the body do you give an IP injection to a rodent?
lower left quadrant of abdomen
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how do you know if you are successful with an IP injection?
- before giving fluid, aspirate
- if nothing aspirates, give fluid
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how much fluid can you give mice with an IP injection?
2 mL
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how much fluid can you give rats with an IP injection?
10 mL
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where is the tail vein located?
laterally on each side of the tail
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where is the tail artery located?
ventral aspect of the tail
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what size needle is used on mice for IV (tail vein) injections?
25-27 ga, 1/2 in.
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what size needle is used on rats for IV (tail vein) injections?
22-25 ga, 1/2 in.
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how do you administer an IV (tail vein) injection on rodents?
- restrain rodent (use restrainer or anesthesia)
- vasodilate tail by warming (hands, heat lamp, heating pad, warm water, hair dryer)
- grasp tail, rotate slightly to see veins
- choose injection site a little over half way down the tail
- DO NOT ASPIRATE
- if vein clears, then you are successful
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what size needle is used on mice for IM injections?
not recommended
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what size needle is used on rats for IM injections?
25 ga, 1/2 in.
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where do you administer an IM injection on rats?
posterior (caudal) thigh muscles
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how much fluid can you give rats with an IM injection?
0.3 mL
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what is the blood volume of a mouse?
70-80 mL/kg or 5.5% of body weight
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what is the blood volume of a rat?
50-65 mL/kg or 6-7% of body weight
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what are the five areas for blood collection in rodents?
- lateral tail vein
- ventral tail artery
- saphenous vein
- retro-orbital venous sinus
- cardiac punctur
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how do you administer blood collection from the lateral tail vein on a mouse?
- restrain mouse
- vasodilate tail vessels
- use scalpel blade to nick the skin and pierce the vein
- collect blood as droplet well up in microtainer or capillary tube
- hold pressure or cauterize when finished
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how much blood can be collected using the lateral tail vein on a mouse?
0.2 mL
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how do you administer blood collection from the lateral tail vein on a rat?
- restrain rat
- vasodilate tail vessels
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how much can be collected using the lateral tail vein on a rat?
1 cc
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what is used to collect the blood from the tail vein on a rat?
25 ga needle with 1 in. syringe or capillary tube
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how do you administer blood collection from the ventral tail artery on a rodent?
same as lateral tail vein
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how much blood can be obtained using the ventral tail artery on a mouse?
0.4 mL
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how much blood can be obtained using the ventral tail artery on a rat?
1.0 mL
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what size needle is used for blood collection from the saphenous vein on a rodent?
20-22 ga
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how do you administer a saphenous vein blood collection on a rodent?
- restrain mouse in centrifuge tube
- anesthetize rat with isoflurane
- shave fur over site on rear leg using clippers
- place small amount of petroleum jelly on site
- use needle to pierce skin and nick the vessel
- collect the blood as droplet wells
- hold pressure or cauterize when finished
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what must be done to a rodent prior to a retro-orbital venous sinus blood collection?
anesthetize rodent
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how do you administer a retro-orbital venous sinus blood collection on a rodent?
- use capillary tube or Pasteur pipette to place pressure on the conjunctiva at the medial canthus (inner corner) of the eye
- twist slightly until the tube pops into the venous sinus
- place a gauze pad over eye, closing eyelids and use gentle pressure to prevent hematomas when finished
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how much blood can be obtained using the retro-orbital venous sinus on a mouse?
0.2 mL
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how much blood can be obtained using the retro-orbital venous sinus on a rat?
0.5 mL
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what must be done to a rodent prior to a cardiac puncture blood collection?
anesthetize rodent
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when do you use a cardiac puncture blood collection on a rodent?
terminal procedure
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how do you administer a cardiac puncture blood collection on a rodent?
- place animal on its back
- insert needle under xiphoid cartilage at 30 degree angle
- aspirate slowly as needle is advanced slightly until blood enters syringe
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what size needle is used on rodents for a cardiac puncture blood collection?
22 ga, 1 in.
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what is an alternate technique for a cardiac puncture?
animal lying on its right side, approach from behind left foreleg
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how much blood can be collected on a mouse using a cardiac puncture?
1.0 mL
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how much blood can be collected on a rat using a cardiac puncture?
6 mL or more
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what is euthanasia?
act of inducing a painless death
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what are the 11 criteria to be considered for euthanasia?
- painless method
- time it takes to produce unconsciousness
- time until death is accomplished
- reliability of method
- repeatability of method
- personnel safety
- stress on animal
- non-reversibility
- compatible with scientific protocol
- aesthetically acceptable
- drug availability and abuse potential
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what are the three methods for minimizing fear and apprehension?
- use gentle restraint and handling
- use tranquilizers or sedatives
- prevent conscious animals from observing the euthanasia of others, especially their own species
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what are the three modes of action of euthanasia methods?
- hypoxemia of brain tissues
- depression of vital brain centers
- physical damage to brain tissue
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what are the two forms of inhalation agents used in euthanasia?
- inhalant anesthetics
- carbon dioxide
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what are examples of inhalation anesthetics used for euthanasia?
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what is the method for using inhalant anesthetics for euthanasia?
- place animal in closed container containing cotton or gauze soaked with agent
- vapors are inhaled
- animal becomes deeply anesthetized
- respiration ceases
- animal expires
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what should not be done using inhalant anesthetics for euthanasia?
- animal come into contact with agent
- air to circulate in the container
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what are the advantages to using inhalant anesthetics for euthanasia?
- good for small animals or for animals that are hard to restrain
- many clinics have anesthesia machines
- most agents are nonflammable and non-explosive
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what are the disadvantages to using inhalant anesthetics for euthanasia?
- some struggling and anxiety may occur
- cannot be used without good ventilation or an anesthetic machine
- may require a secondary method
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what is the method for using carbon dioxide for euthanasia?
- place animal in chamber filled with carbon dioxide
- animal loses consciousness due to anesthetizing effect
- animal asphyxiates
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what are the advantages for using carbon dioxide for euthanasia?
- inexpensive
- nonflammable
- non-explosive
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what are the disadvantages for using carbon dioxide for euthanasia?
cannot be used on neonatal animals
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what are the two forms of injectable agents used for euthanasia?
- barbiturates
- potassium chloride
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what are examples of barbiturates used for euthanasia?
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what is the method for using barbiturates for euthanasia?
intravenous in most animals
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what barbiturate must be given IV?
thiopental
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what barbiturate can be given IP to rodents?
pentobarbital
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what are the advantages to using barbiturates for euthanasia?
- fast
- minimal discomfort to the animal in most cases
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what are the disadvantages to using barbiturates for euthanasia?
- necessitates trained personnel to restrain animal and give injection
- regulated - must have DEA number
- terminal gasp is aesthetically objectionable
- excitability is occasionally possible as animal passes through stages of anesthesia
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what is the method for using potassium chloride for euthanasia?
- IV only in DEEPLY anesthetized animals
- (be careful adding too much to IV fluids as is harmful)
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what are the advantages to using potassium chloride for euthanasia?
- inexpensive
- fast
- non-regulated
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what are the disadvantages to using potassium chloride for euthanasia?
- animal must be anesthetized
- painful if animal not deeply anesthetized
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what is the form of physical euthanasia?
captive bolt pistol
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what is the method to using captive bolt pistol for euthanasia?
- animal is restrained
- pistol is placed between the the eyes and perpendicular to the skull and then discharged
- bolt penetrates the skull and significantly damages the brain to cause unconsciousness
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what are the advantages to using captive bolt pistol for euthanasia?
humane method for large animals that cannot be euthanized with drugs if used correctly
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what are the disadvantages to using captive bolt pistol for euthanasia?
- aesthetically displeasing
- death may not occur
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what are the three form of secondary methods to ensure death?
- cervical dislocation
- exsanguination
- rapid freezing
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what is the method for using cervical dislocation?
- after animal in anesthetized, place thumb and index finger on either side of the neck at the base of the skull
- using the other hand, the base of the tail is quickly pulled to separate the neck vertebrae
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what is the method for using exsanguination?
remove 75% of blood volume after the animal has been anesthetized
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what is the method for using rapid freezing?
liquid nitrogen is used to freeze brain tissue
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when is rapid freezing an acceptable form of euthanasia?
brain biochemistry is important for research project
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when is exsanguination an acceptable form of euthanasia?
- large volumes of blood are needed
- tissues must be used for histopathology or other tests
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which euthanasia method is suitable for dogs?
injectable barbiturates
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which euthanasia method is suitable for cats?
injectable barbiturates
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which euthanasia method is suitable for cows?
- injectable barbiturates
- captive bolt pistol
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which euthanasia method is suitable for horses?
- injectable barbiturates
- captive bolt pistol
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which euthanasia method is suitable for sheep?
- injectable barbiturates
- captive bolt pistol
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which euthanasia method is suitable for mice?
- carbon dioxide
- injectable barbiturates
- potassium chloride
- cervical dislocation (under 200 g)
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which euthanasia method is suitable for rats?
- carbon dioxide
- injectable barbiturates
- potassium chloride
- cervical dislocation (under 200 g)
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which euthanasia method is suitable for birds?
- inhalant anesthetics
- cervical dislocation
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which euthanasia method is suitable for rabbits?
- carbon dioxide
- injectable barbiturates
- potassium chloride
- cervical dislocation (under 1 kg)
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what are the four variables that will affect the choice of method for euthanasia?
- availability of drugs
- personnel qualifications
- animal species
- experimental focus
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