Guidelines for Anesthesia, Analgesia, and Tranquilization

Policy No.: 104.01

Revision No: 4
Effective Date: (revised

02/25/14)

Category: Research Guidelines

I. GENERAL

2
The Animal Welfare Act and the Public Health Service Policy on Humane Care
and Use of Laboratory Animals requires that with all procedures in which the
animal perceives pain, the investigator minimizes or alleviates that pain or stress
or provides justification that alleviation of the pain would interfere with goals of
the experimental design.
3

II. SELECTION OF THE APPROPRIATE DRUG

4
There are several considerations in the selection of a particular drug for an
experimental procedure.
The purpose of the drug (restraint, blood collection, dosing, or surgical
anesthesia) will dictate the use of one type of agent over another.
The type of study or procedure will require a drug which interferes the least with
the experimental parameters being observed.
The requirement of a post-operative analgesic may indicate the use of an
anesthetic which inherently provides analgesia (such as Innovar-Vet).
The available equipment or facilities (such an inhalation equipment, scavenging
devices, or hood) may dictate the use of one anesthesia over another to
accommodate the facility.
The use of drugs to alleviate stress and/or pain in experimental procedures may
affect several parameters including the cardiovascular system, respiration, and
thermoregulation in addition to the central nervous system. It is important in
administration of such agents to monitor physiological parameters and maintain
acceptable levels. A commonly employed device for maintaining an animal's
temperature during anesthesia is a warm water heating pad. To maintain normal
hydration, IV or SC fluids are administered.
The experience of the individual designated to administer the drug is another
important factor. Some anesthesias have a wide safety margin and can be used
safely by relatively inexperienced individuals. Other agents require experienced,
knowledgeable personnel to administer them due to the narrow safety margin
and possible complications of their use.

III. COMMONLY USED DRUGS

Tranquilizers, sedatives, and hypnotics
The purpose of these agents is to calm anxiety. Generally they do not
provide analgesia or a loss of consciousness. Many tranquilizers are
frequently used as preanesthetics in veterinary medicine with the dual
purpose of calming the patient and lowering the total anesthetic required
to produce anesthesia. Some examples of commonly used veterinary
tranquilizers or preanesthetics are:
Acetylpromazine or "Ace" is a potent neuroleptic for many animals
including wild species. "Ace" has many advantages as a tranquilizer:
wide safety margin, low toxicity, potent sedative action, good muscle
relaxation, and can be used in combination with narcotics, dissociative
anesthetics, and atropine. Ace is also a potent vasodilator, thus
hypotension can be a concern.
Diazepam or Valium is a mild sedative with muscle relaxant and
anticonvulsant properties. It is an excellent preanesthetic for animals with
seizure disorders. Some formulations must be administered intravenously
and cannot be physically mixed with atropine as precipitation results.
Care must be taken to administer the drug slowly if administered IV.
Xylazine or Rompun is not a true tranquilizer due to its property of
providing analgesia as well as muscle relaxation. It is used as a
preanesthetic sedative. When used with a barbiturate, xylazine will
reduce the total anesthesia required for the procedure. Xylazine can be
used in combination with ketamine, ultrashort-acting barbiturates,
halothane, or methoxyflurane. Xylazine can cause bradycardia and
should be used only after premedication with atropine or glycopyrrolate.
Increasing dosages increases the duration of its effects rather than the
degree of sedation. Dosages vary widely between species.
Analgesics
Analgesic agents are used to alleviate or reduce pain. The minimizing of
stress and the use of analgesia to alleviate pain is an important
consideration in a research protocol. In the conduct of humane research
it is important that the scientist and staff be familiar with signs exhibited by
the animal which would indicate pain. Below are two tables from Lab
Animal, vol. 15, #9, page 32 to assist in the evaluation of pain in rodents.
The signs described may be extended to other species.

Table 1. Signs for

judging morbidity
(disease/illness in
rodents)

Table 2. Signs for judging moribund

condition in rodents

Rapid breathing
rate
Breathing rate very
slow, shallow, and
labored
Rapid weight loss
Ruffled fur (rough
hair coat)
Hunched posture
Hypothermia or
hyperthermia
Ulcerative
dermatitis or
infected tumors
Inappetence

Impaired ambulation (unable to reach food

or water easily)
Evidence of muscle atrophy or other signs
of emaciation (body weight is not always
proportionate)
Any obvious illness including such signs
as lethargy (drowsiness, aversion to
activity, lack of physical or mental
alertness), prolonged incontinence,
bleeding, difficulty breathing, central
nervous system disturbances, or chronic
diarrhea or constipation
Inability to remain upright

Diarrhea or
constipation
Below is a table appearing in Lab Animal, vol. 14, #4, page 25 which
describes the potential for pain in various experimental manipulations.
Table 1. Post-Operative Pain Evaluation
Type of Surgery

Head, ear,
throat, dental

Ophthalmologic

Orthopedic

Abdominal
Cardiovascular

Indications of Pain
Response
Rubbing, shaking, selfmutilation, depression,
and reluctance to
swallow, eat, drink or
move
Reluctance to move,
scratching, rubbing
Abnormal posture,
abnormal gait,
reluctance to move,
guarding, itching, selfmutilation
Guarding, abnormal
posture, vomiting,
anorexia
Depression, reluctance

Expected
Pain Level

Duration

Moderate
to high

Generally
intermittent

High

Intermittent
to continual

Moderate

Intermittent

Mild to
moderate

Intermittent

Mild to

Continual

to move, anxiety
Changes in respiratory
rate and pattern, anxiety,
reluctance to move
Scooting, biting, licking,
self-mutilation

Thoracic
Perirectal

moderate
Mild to
moderate

Continual

Moderate
to high

Intermittent
to continual

Some examples of commonly used analgesic agents are discussed

below.
Narcotics and Opiates effectively raise the pain threshold as well
as altering the psychological response to pain. They provide both
sedation and strong analgesia. Narcotics induce a "sleep-like"
state, or stupor, through CNS depression. Disadvantages to their
use are the depressant action on the CNS, cardiovascular, and
respirator systems. Dosages vary considerably between species,
necessitating confirmation of a suitable dose with the clinical
veterinarian. Commonly used narcotics in veterinary applications
are:
1

Buprenorphine is a derivative of the morphine alkaloid,

thebaine. It has profound analgesic properties when
administered parenterally which last 6-12 hours. It is
suitable for relief of moderate to severe pain in multiple
species.

Buprenorphine SR is a sustained release

(SR) formulation that delivers buprenorphine in
a fully biodegradable liquid polymer matrix.
This formulation provides consistent 72 hour
release of buprenorphine. Studies have been
done in dogs, cats and laboratory animals that
have shown that over the 72 hour period blood
levels are greater than 1 nanogram/mL for the
entire 72 hour period. A study in macaques
showed this plasma concentration was
maintained for 5 days.

Butorphonol Tartrate is an opioid agonistantagonist which is suitable for controlling

moderate to severe pain in several species
including cats. Its duration of action is 4-6
hours. It causes less respiratory depression
than morphine.
Codeine is a less effective analgesic in animals than
morphine and is rarely used. It can be used to control
irritating coughs in dogs.

Fentanyl is a short-acting analgesic. Effects of fentanyl

are similar to those of morphine, however, respiratory

center depression can be more severe and apnea may

result.
5

Meperidine (Demoral) is one fifth as potent as morphine.

Meperidine may be used in dogs, nonhuman primates, and
horses. It is recommended for alleviation of short-duration
post-operative pain or in situations which require repeated
drug administration to control pain. Rapid IV injection may
cause convulsions.

Morphine is used in dogs and nonhuman primates.

Adverse reactions are seen in cats, horses, and cattle
including increased spinal reflexes, panting, and nausea.
Nonhuman primates are reported to exhibit skin irritation
on the palms and soles of their feet and a loss of appetite.
Depression of gastrointestinal tract mobility is a common
side effect of morphine. Morphine depresses the
thermoregulatory center and animals should be maintained
in a temperature controlled environment when undergoing
morphine treatment. Morphine causes an increase in
intraocular pressure in most species and pupillary
constriction in dogs and rabbits. Duration of action of
morphine is 2-4 hours.

Oxymorphone as an analgesic is ten times more potent

than morphine with a less depressant effect on the
respiratory center and less stimulation of vomiting.
Oxymorphone can cause bradycardia and auditory
hypersensitivity.

Pentazocine is one third as potent as morphine with mild

respiratory and CNS depression. Duration of action is
about one hour. Generally, it is not considered a potent
analgesia and is used to control chronic or mild postsurgical pain.
Tramadol Hydrochloride is an oral opioid that is currently
not a scheduled drug. Although its mode of action is not
completely understood at least two mechanisms seem to
be applicable: binding of the parent and the M1 metabolite
to u-opioid receptors and weak inhibition of reuptake of
norepinephrine and serotonin.

Non-Narcotic Analgesics, also classified as non-steroidal antiinflammatory agents (NSAID) generally mediate the inflammatory
response through antiprostaglandins or prostaglandin synthesis
inhibitors.
Carprofen (Rimadyl) has antinflammatory, analgesic,
and antipyretic activity similar to other NSAIDs. It is
considered to be COX-2 specific. In dogs it is given twice
daily.

Deracoxib (Deramaxx) has antinflammatory, analgesic,

and antipyretic activity similar to other NSAIDs. It is
considered to be COX-2 specific. It is newer than
carprofen and can be given to dogs once daily.
Flunixin meglumine (Banamine) is a potent injectable
nonsteroidal anti-inflammatory agent with potent analgesic
and fever-reducing properties. It must be used cautiously
for no more than 3 days consecutively.

Meloxicam (Metacam) has antinflammatory, analgesic,

and antipyretic activity similar to other NSAIDs. It is
considered to be COX-2 preferential, not COX-2 specific,
as at higher dosages its COX-2 specificity is diminished.

Saliscylates and Acetaminophens include both aspirin

and Tylenol which are effective against superficial pain,
especially that of musculoskeletal origin. Acetaminophen
is excessively toxic to cats due to their inability to detoxify
the equivalent of a human dose.

General Anesthesia
General anesthesia is a state of complete unconsciousness with the
following components: loss of pain perception, loss of motor control,
skeletal muscle relaxation, and absence of reflex response. The
unconscious state cannot be interrupted by stimulation as can the
narcotic induced stupor. Termination of the unconscious state of general
anesthesia is accompanied by pain perception. General anesthesia can
be administered by inhalation or injection methods. Common injectable
anesthetics are:
1

Barbiturates are a group of drugs with similar chemical

structures. Intermediate and long duration barbiturates are used
as hypnotics, sedatives, or anti-convulsants. Barbiturates act as
CNS depressants and are considered poor analgesics. Shortacting or ultra-short acting are generally acceptable as anesthetic
agents.
Increasing barbiturate doses will progressively increase the level
of CNS depression until a state of anesthesia is reached. They
are potent respiratory center depressants and can have variable
effects on the cardiovascular system. There is considerable
variation in dose response within and between species.
Barbiturates should be administered intravenously, slowly to
effect. Other methods provide less predictable effects. The
administration of glucose, chloramphenicol and epinephrine will all
prolong recovery from barbiturate anesthesia.

Dissociative Anesthetics produce an anesthetized state

characterized by moderate analgesia, muscle rigidity, loss of
responsiveness to external physical stimuli, and amnesia. Eyes
remain open and various reflexes remain intact (blinking),

laryngeal reflex. The respiratory center is not depressed and

frequently there is an increase in heart rate and blood pressure.
Ketamine hydrochloride is the commonly used dissociate
anesthetic in veterinary medicine. In some species, such
as cats and dogs, side effects of ketamine anesthesia
include: excessive salivation, muscle rigidity, a tendency
toward convulsions, and recovery characterized by
excitement, disorientation, and hallucinations. The extent
of analgesia provided for invasive procedures is
questionable and Ketamine should not be used alone in
these cases. The use of narcotic preanesthetics is
recommended for the reduction of the side effects of
ketamine.
Telazol is a combination injectable anesthetic composed of
tiletamine, a dissociative anesthetic related to ketamine,
and zolazepam, a water soluble benzodiazepine
tranquilizer. It is suitable for short noninvasive procedures
in some species including primates and swine, and may be
used for injection prior to general gas anesthesia.
Tribromoethanol (Avertin)
Tribromoethanol (Avertin) is an anesthetic that provides
rapid induction and recovery for single use, short duration
(approximately 15-20 minutes) surgical procedures in
rodents. Tribromoethanol has been commonly used in the
production of transgenic animals to facilitate procedures
such as embryo transfer, vasectomy, or distal tail
amputation for Southern Blot analysis.
Tribromoethanol is not commercially available and
investigators who wish to use it as an anesthetic must
make their own solutions. Tribromoethanol may be
approved for use after scientific and/or medical justification
has been provided in the animal care and use protocol as
to why a pharmaceutical grade anesthetic agent cannot be
used. TJU strongly encourages the use of
ketamine/xylazine or isoflurane rather than
Tribromoethanol.
Inhalation Anesthesia
The use of inhalant anesthesia provides the user with several
advantages over injectable anesthesia. The rapid induction and
recovery as well as the overall controllability of the anesthesia
provide a greater means of absolute control of the animal's
physiological state. However, inhalant anesthesia requires
specialized equipment, constant monitoring, and trained technical
assistance. Equipment can be a simplistic nose cone for
administration to rodents or complex apparatus required for larger
species. Several devices have been designed for the
administration of inhalant anesthesia to the variety of species

commonly used in the laboratory and consultation of the literature

may be helpful.
Commonly used inhalant anesthetics are:
Isoflurane is an anesthetic with almost "ideal" properties.
It is non-flammable, non-irritating, non-toxic, and relatively
insoluble in the blood. It has rapid induction and recovery.
Isoflurane is encouraged as the first choice anesthetic in
mice. It should be delivered as a known percentage (1-3%
for maintenance; up to 5% for induction) in oxygen from a
precision vaporizer.
Sevoflurane is an inhalation anesthetic similar to
isoflurane, but has a very rapid anesthesia induction and
recovery.
Methoxyflurane is a potent anesthetic which provides
significant post-operative analgesia. Its low volatility allows
its use with a simple nose cone for small rodents. Muscle
relaxation is good and it is neither explosive nor irritating in
anesthetic doses. Renal and hepatic damage is possible
with prolonged or excessive doses.
Halothane is a widely used inhalant anesthetic in
veterinary medicine. It provides rapid induction and
recovery, fair muscle relaxation, and poor analgesia.
Halothane is highly volatile and should be used with a
finely-calibrated precision vaporizer. It may induce
malignant hyperthermia in certain breeds of swine.
Nitrous Oxide is a weak anesthetic drug for veterinary use
and must not be used by itself. It can be used both with
other inhalant anesthetics or narcotic analgesia. Used in
combination with other inhalant anesthetics it reduces the
dose needed of these agents.
Diethyl Ether is a highly volatile anesthetic agent. Ether is
not an approved anesthetic agent at Thomas Jefferson
University.
A precision vaporizer must be used for delivering inhalant
anesthetics. Use of a cotton ball or gauze soaked with the
anesthetic in a closed container is strongly discouraged
and can only be used if in an approved protocol. Strong
justification will be required before approval is granted.
Anticholinergic Drugs
The use of anticholinergic drugs such as atropine or
glycopyrrolate reduce salivation, lacrimation, urination, and
defecation during the experimental procedure. Atropine is the
most commonly used anticholinergic. It should be noted that

rabbits have the enzyme atropinesterase which results in a

decreased response to the administration of atropine.
GUIDELINES FOR ANESTHESIA, ANALGESIA, AND TRANQUILIZATION
1

o Cats

o Dogs

o Rats

o Mice

o Hamsters

o Gerbils

o Rabbits

o Guinea Pigs

o Primates

10 o Pigs
11 o Sheep & Goats

Guidelines for Anesthesia, Analgesia, and Tranquilization: Tables

CAT
DRUG
Acepromazine
(tranquilization)
Atropine
Buprenorphine

ANESTHESIA
Premeds
0.03-0.05 mg/kg IV,
IM, SC (2 mg max).
0.02-0.04 mg/kg IV,
IM, SC
0.01 - 0.02 mg/kg
IV, IM, SC

ANALGESIA

Induction Agent

0.005-0.01 mg/kg IV,

IM, SC q6-12h

Buprenorphine
SR

0.12 mg/kg SC q72h

Butorphanol

0.2 - 0.4 mg/kg IV,

IM, SC

0.2 0.4 mg/kg IM,

SC q2-4h; tablets:
0.4 mg/kq q8h

Diazepam
(tranquilization)

0.1-0.2 mg/kg IV

**1.25-2.5 mg patch
(12.5-25 mcg/hr);
duration at least 72
hrs and is longer in
cats than dogs.

0.1-0.2 mg/kg IV, IM,

Fentanyl

Glycopyrrolate
Hydromorphone

0.01-0.02 mg/kg IV,

IM, SC
0.1-0.2 mg/kg IV,

Isoflurane

Ketamine

Ketamine +
Diazepam

Meloxicam
(NSAID)

Meperidine
Midazolam
(tranquilization)

IM, SC
1-3% inhalant to
effect (up to 5% for
induction)
22-33 mg/kg can
be used for dx or
mionor sx
procedures, rec.
using with xylazine
or diazepam

SC q 2-6h

2-6 mg/kg IV, IM,

SC
0.06-0.2 mg/kg
with opioid IV, IM,
SC

Morphine

Oxymorphone

K: 5-10 mg/kg IV
D: 0.05 mg/kg IV

0.2 mg/kg PO, SQ

loading dose once,
followed by 0.1
mg/kg PO, SQ q 24h
for not longer than 2
days.
3-5 mg/kg IM, SC q12h

0.1 mg/kg SC q1224h

0.05-0.1 mg/kg SC
q1-3h

1-2mg/kg PO q8-12h

Propofol
Telazol
Thiopental
Tramadol

4-8 mg/kg IV

1-4 mg IV, 4-8 mg IM

8-10 mg/kg IV

0.5-2 mg/kg IV, IM,

Xylazine

SQ
** Small cats may be dosed with patch, but the patch should not be cut in half. Cover
of the gel membrane with tape. Half-patch dosing is suggested for pediatric,
geriatric, and systemically ill cats.
DOG
DRUG
Aspirin
Acepromazine
(tranquilization)
Atropine

ANESTHESIA
Premeds
...
0.03-0.05 mg/kg IV,
IM, SC (2 mg max).
0.02-0.04 mg/kg IV,
IM, SQ

ANALGESIA

Induction Agent

10-20 mg/kg q12h

Buprenorphine

0.01 - 0.02 mg/kg

IV, IM, SC

Buprenorphine
SR

Butorphanol

0.2 - 0.4 mg/kg IV,

IM, SC

Carprofen
(NSAID)

2 mg/kg PO q12h

Codeine

0.5-2 mg/kg PO q612h

1-2 mg/kg PO q24h

0.1-0.2 mg/kg IV

Deracoxib
(NSAID)
Diazepam
(tranquilization)
Fentanyl
Glycopyrrolate
Hydromorphone
Isoflurane
Ketamine +
Diazepam
Meloxicam
(NSAID)
Meperidine
Midazolam
(tranquilization)

0.01-0.02 mg/kg IV,

IM, SC
0.1-0.2 mg/kg IV,
IM, SC
1-3% inhalant to
effect (up to 5% for
induction)
D: 0.5 mg/kg IV
then ketamine
K: 10 mg/kg IV

2-6 mg/kg IV, IM,

SC
0.06-0.2 mg/kg
with opioid IV, IM,
SC

Morphine

0.1-2 mg/kg IM, SC

Oxymorphone

Pentobarbital
Propofol
Thiopental
Tramadol

30 mg/kg IV

0.005-0.02 mg/kg IV,

IM, SC q6-12h
0.03-0.06 mg/kg SC
q 72h
0.2 1 mg/kg IM, SC
q2-4h; tablets: 0.4
mg/kq q8h

See below

0.1-0.2 mg/kg IV, IM,

SC q 2-6h

0.2 mg/kg PO, SQ

loading dose once,
followed by 0.1
mg/kg PO, SC q 24h
3-5 mg/kg IM, SC
q1-2h

0.5-2 mg/kg IM, SC

q3-4h; tablets: 1.5-3
mg/kg PO q12h
0.22 mg/kg SC q 13h

1-5 mg/kg PO q8-12h

4-8 mg/kg IV
8-10 mg/kg IV

Xylazine

0.5-2 mg/kg IV, IM,

SC

DOG Fentanyl
Canine Size
Dose
Fentanyl Content
< 5 kg **
12.5- 25 mcg/hr
1.25-2.5 mg
5-10 kg
25 mcg/hr
2.5 mg
10-20 kg
50 mcg/hr
5 mg
20-30 kg
75 mcg/hr
7.5 mg
> 30 kg
100 mcg/hr
10 mg
** Small dogs may be dosed with patch, but the patch should not be cut in half. Cover
of the gel membrane with tape. Half-patch dosing is suggested for pediatric,
geriatric, and systemically ill dogs.

RAT & MOUSE: Ketamine, Acepromazine, and Xylazine cocktail Atropine.

It is strongly recommended that atropine be administered (as a separate injection) due to
the cardiovascular depressant effects of the following mixture.
In one bottle of ketamine (100 mg/ml) add two ml of acepromazine (10 mg/ml) and 0.5
ml of xylazine (100 mg/ml).
* Please note that the 100 mg/ml bottle of xylazine is used, not the 20 mg/ml bottle.

This will give final concentrations of 80 mg/ml of ketamine, 1.6 mg/ml of acepromazine,
and 5 mg/ml of xylazine. Dose the animal at 1.25 mls/kg of body weight or 0.125
mls/100 grams of body weight subcutaneously (works nicely using the SC route, can
probably also be administered IP).
Atropine: Given at approximately 0.05 mg/kg mix 0.25 mls of atropine in 4.75 mls of
normal saline. This will give a final concentration of 0.027 mg/ml. Dose the animal at
0.054 mg/kg or 0.2 mls/100 grams of body weight subcutaneously.
The recommended shelf life for the ketamine, xylazine, aceproazine cocktail is 180 days
after mixing, provided that any of the 3 drugs in the mixture have not expired before that
time.

Taylor, B. et al. 2009. Beyond-Use Dating of Extemporaneously Compounded Ketamine,

Acepromazine, and Xylazine: Safety, Stability, and Efficacy Over Time. JAALAS 48(6):
718-726.

Atropine in saline should be discarded immediately after use.

Rat & Mice Tribromoethanol (Avertin)

Improper preparation, storage, or use of Tribromoethanol can result in high mortality
losses. In particular, Tribromoethanol degrades in the presence of heat and light,
producing toxic by-products that are potent gastrointestinal irritants.

The procedure for production, storage and use of Avertin was provided by the
Jackson Laboratory, where it is used routinely for mouse surgery.

Preparation of Tribromoethanol Stock Solution

Items Needed:
1ml syringes
10ml syringes
100ml wide mouth glass bottle
(2) amber vials with silicone septa, 40ml
50g 2,2,2 tribromoethanol powder
tert-amyl alcohol
25mm syringe filter with 45um membrane
pH paper (range 4.0 to 7.0)
Tribromoethanol stock labels
Fisherbrand marking pen
Clean lab coat
Gloves
NOTE: Glass bottle and amber vials must be cleaned and steam sterilized prior to use.
Preparation of stock solution
1. In a fume hood add 31mls tert-amyl alcohol to 50g of 2,2,2tribromoethanol powder in a 100ml wide mouth glass bottle. Cap the
bottle tightly and manually shake for a few minutes. This should get
most of the powder into solution.

2. Place the bottle in a 600C shaker water bath. Gently shake the bottle
until all of the crystals are dissolved.
3. Once dissolved test the pH of the stock solution. It should be between 5.0 and 5.5.
4. In a laminar flow clean bench use 10ml syringes to withdraw the tribromoethanol
stock solution from the bottle.
5. Attach a syringe filter to a 10ml syringe and filter the tribromoethanol stock solution
directly into the 40ml amber vial. Use a new syringe and syringe filter for each 10mls
of stock solution.
6. Repeat Steps 4-5 until all the stock solution is removed from the bottle.
7. Place a stock label on the bottle. Record on the label: chemical name, preparation
date, expiration date, who prepared the stock solution, and bottle number. NOTE:
There is 1-month expiration date for tribromoethanol stock solution.
8. Store stock solution in dark at room temperature.
Preparation of the working solution of tribromoethanol (20mg/ml)
Items Needed:
Amber vials with silicone septa, filled with 39.5mls 1X PBS
Tribromoethanol stock solution
1ml syringes
20 gauge needles
70% ethanol
pH paper (range 4.0 to 7.0)
Fisherbrand marking pen
Clean lab coat
Gloves
NOTE: Amber vials must be cleaned and steam sterilized prior to use.
Preparation of working solution
1. Place the vials containing PBS into a water bath, bring up to 500C and heat for 10
minutes.
2. In a laminar flow clean bench fill 1ml syringes with tribromoethanol stock solution.
3. After 10 minutes at 500C remove and dry vials and place in the laminar flow clean
bench.
4. Disinfect the top of each vial with 70% ethanol.
5. Inject 0.5ml of stock solution into each vial while the PBS is still warm and manually
shake each vial until the solution is thoroughly mixed (no bubbles/crystals visible).
6. Test the pH of one vial in the batch. The pH should be 7.0.
9. Place a 2% tribromoethanol label on the bottle. Record on the label: chemical name,
preparation date, expiration date, who prepared the stock solution, and bottle
number. NOTE: There is a 3-week expiration date for tribromoethanol working
solution.
10. Store working solution in dark at room temperature.
Dosage:
The dosage for the C57BL/6J mouse is ~0.2 ml/10 gram body weight (400 mg/kg) IP.
Dosage will need to be adjusted by mouse strain.

RATS
DRUG
Acepromazine

ANESTHESIA

ANALGESIA

200 - 300 mg/kg

PO
1-2 mg/ml in
drinking water
100-150 mg/kg
PO q4h

0.1-0.5 mg/kg
SC, IP q8-12h
0.9-1.2 mg/kg
SC q72h
0.2-2 mg/kg SC,
IP q2-4h

1.1-2.5 mg/kg
SC q12h

TRANQUILIZATION
0.5-1 mg/
kg IP, SC

Acetaminophen
(NSAID)

Aspirin (NSAID)

Atropine (premed)

0.1-0.4 mg/kg SC

Buprenorphine

Buprenorphine SR

Butorphanol

Diazepam

Flunixin (NSAID)

Glycopyrrolate
(premed)

0.01-0.02 mg/kg SC

Ibuprofen (NSAID)

10-30 mg/kg
PO q4h

Isoflurane
Ketamine +
acetylpromazine

1-3% inhalant to
effect (up to 5% for
induction)
K: 75 mg/kg IP, SC
A: 2.5 mg/kg IP, SC

4.0 mg/kg SC

Ketamine +
Diazepam

K: 60 - 80 mg/kg IP,
SC
D: 10 mg/kg IP, SC

Ketamine +
Xylazine +
Acepromazine

See description
above.

Ketamine +
Xylazine

K: 40 - 80 mg/kg IP,
SC
X: 5 - 10 mg/kg IP,
SC

Meloxicam (NSAID)

Meperidine

Midazolam

Morphine

Oxymorphone

Pentobarbital

30-45 mg/kg IP

0.2-0.3 mg/kg
PO, SC q12h
10-20 mg/kg SC
q2-3h

2-5 mg/kg SC
q2-4h
0.2-0.5 mg/kg
SCq6-12h

1-2 mg/kg SC

* Different strains and stocks may react differently to drug combinations.

MICE
DRUG
Acepromazine
Atropine (premed)

ANESTHESIA

0.05 - 0.1 mg/kg SC

Acetaminophen
(NSAID)

Aspirin (NSAID)

Buprenorphine

..

Buprenorphine SR

Butorphanol

Carprofen (NSAID)
Diazepam

Flunixin (NSAID)

ANALGESIA

300 mg/kg PO
1-2 mg/ml in
drinking water
120 mg/kg PO q
4h
0.05-2.5 mg/kg
SC, IP q6-12h
0.3 0.6 mg/kg SC
q72h
1-5 mg/kg SC q
4h
5mg/kg SC q24h

0.3 -2 mg/kg SC,

Po q12-24h

TRANQUILIZATION
0.5-1.0 mg/kg IP, SC

3-5 mg/kg IP

Glycopyrrolate
(premed)

0.01-0.02 mg/kg SC

Ibuprofen (NSAID)

..

7-15 mg/kg PO
q4hr

Ketamein +
Xylazine

1-3% inhalant to effect

(up to 5% for
induction)
K: 50 mg/kg IP, SC
X: 5 mg/kg IP, SC

Meloxicam (NSAID)

0.2-0.3 mg/kg
PO, SQ q 12hrs

Ketamine + Xylazine
+ Acepromazine

See description
above.

Meperidine

Morphine

Oxymorphone

..

Pentobarbital

50-90 mg/kg IP, SC

Isoflurane

20 mg/kg Sc q23h
2-5 mg/kg SC q2-4h
0.2-0.5 mg/kg
SC q6-12h

* Different strains and stocks may react differently to drug combinations.

HAMSTERS
DRUG
Acepromazine
Acetaminophen
(NSAID)
Atropine (premed)

ANESTHESIA

ANALGESIA

1-2 mg/ml in
water
...
0.5 mg/kg SC
q8h
1-5 mg/kg SC
q4h

TRANQUILIZATION
0.5-1.0 mg/kg IP, SC

5 mg/kg SC q24h

2.5 mg/kg SC
q12-24h

3-5 mg/kg IP

0.05-0.1 mg/kg SC

Buprenorphine

Butorphanol

Carporfen
(NSAID)
Diazepam
Flunixin (NSAID)

Glycopyrrolate
(premed)

0.01-0.02 mg/kg SC

Isoflurane
Ketamine +
Xylazine

1-3% inhalant to effect

(up to 5% for induction)
K: 80 mg/kg IP
X: 5 mg/kg IP

...

Meloxicam
(NSAID)

Meperidine

Morphine

Oxymorphone

Pentobarbital

50-90 mg/kg IP

0.2-0.3 mg/kg
PO, SC q12h
20 mg/kg SC q23h
2-5 mg/kg SC
q8h
0.2-0.5 mg/kg SC
q6-12h

Hamsters exhibit a great variability of response to administration of parenteral

anesthesics.
The use of droperidol and fentanyl may result in CNS signs and should be avoided if
possible.
Intravenous injections may be made into the saphenous vein.
GERBILS
DRUG
Acetaminophen
(NSAID)
Atropine (premed)

Buprenorphine

Butorphanol

Carprofen
(NSAID)
Diazepam

ANESTHESIA

0.05-0.1 mg/kg SC

5 mg/kg SC q24h

...
2.5 mg/kg SC
q12-24h

3-5 mg/kg IP

Glycopyrrolate
(premed)

0.01-0.02 mg/kg SQ

Ketamine +
Xylazine
Meloxicam
(NSAID)

TRANQUILIZATION

Flunixin (NSAID)

Isoflurane

ANALGESIA
1-2 mg/ml in
water

0.1-0.2 mg/kg SC
q8h
1-5 mg/kg SC
q4h

1-3% inhalant to effect

(up to 5% for induction)
K: 50 mg/kg IP, SC
X: 2 mg/kg IP, SC

Meperidine

Morphine

Oxymorphone

0.2-0.3 mg/kg
PO, SC q12h
20 mg/kg SC q23h
2-5 mg/kg SC
q8h
0.2-0.5 mg/kg SC
q6-12h

Pentobarbital

50-90 mg/kg IP

In gerbils Acepromazine can cause seizures.

RABBITS
DRUG
Acepromazine

Aspirin (NSAID)

Buprenorphine

Buprenorphine
SR

Butorphanol

Carporfen
(NSAID)

Diazepam

Fentanyl

Flunixin (NSAID)

Glycopyrrolate
(premed)

0.01-0.02 mg/kg SC

Isoflurane
Ketamine +
Acepromazine
Ketamine +
Diazepam
Ketamine +
Xylazine
Meloxicam
(NSAID)

ANESTHESIA

1-3% inhalant to effect

(up to 5% for induction)
K: 25-40 mg/kg IM, SC
A: 0.25-1 mg/kg IM, SC
K: 30-40 mg/kg IM, SC
D: 2-5 mg/kg IM, SC
K: 30-40 mg/kg IM, SC
X: 5 mg/kg IM, SC

ANALGESIA

100 mg/kg PO
q24h
0.02-0.1 mg/kg SC
q6-12h
0.1-0.3 mg/kg SC
q72h
0.1-0.5 mg/kg IM,
SQ q4h
1-2 mg/kg
POq12h; 2-4
mg/kg PO q24h

2.5 mg patch (25

mcg/hr) x 3 days
1-2 mg/kg SC q1224h

TRANQUILIZATION
1-2 mg/kg SQ, IM

1-5 mg/kg IM

Meperidine

Midazolam

0.2-0.3 mg/kg PO,

SC q12-24h
5-10 mg/kg SC, IM
q2-3h

Morphine

2-5 mg/kg IM, SC

q4h

Pentobarbital

20 - 45 mg/kg IV (pre
anesthetize with 1
mg/kg Ace IM

1-2 mg/kg IM

Intramuscular injections of ketamine have been associated with local necrosis and distal
neuropathies in the injected leg.

GUINEA PIGS
DRUG
Acepromazine
Aspirin (NSAID)
Atropine
(premed)

ANESTHESIA
...
...

...
80-85mg/kg PO q4h

TRANQUILIZATION
0.5-1 mg/kg IM
...

0.1-0.2 mg/kg SC, IM

Buprenorphine

...

Butorphanol

Carprofen
(NSAID)

Diazepam
Glycopyrrolate
(premed)
Ibuprofen
Isoflurane
Ketamine +
Acepromazine
Ketamine +
Diazepam
Ketamine +
Xylazine
Meloxicam
(NSAID)

...
0.01-0.02 mg/kg IM,
SQ
...
1-3% inhalant to effect
(up to 5% for
induction)
K: 20-40 mg/kg
A: 0.5 mg/kg IM
K: 20-30 mg/kg IM
D: 1-2 mg/kg IM
K: 20-40 mg/kg IM
X: 2 mg/kg IM

Meperidine

...

Morphine

...

ANALGESIA

0.05-0.10 mg/kg SC
q6-12h
0.4-2 mg/kg SC q24h
1-2 mg/kg PO q1224h; 4 mg/kg SC
q24h
...

...

0.5-3 mg/kg IM

10 mg/kg PO q4h

...

...

...

...

...

...

0.2-0.3 mg/kg PO
q12h
10-20 mg/kg SC,IM
q2-3h
5-10 mg/kg SC,IM
q4h

...
...

Guinea pigs are an anesthetic challenge. They lack accessible blood vessels for IV
administration and display great variability in response. It is not uncommon for two
guinea pigs of the same sex, age, weight, and strain to respond quite differently to the
same anesthesia. Guinea pigs can also exhibit a swimming motion with their legs during
the surgical plane of anesthesia.

Halothane is considered high risk for guinea pigs as they hold their breath and then
gasp. Halothane (1%) causes a 35% - 40% drop in blood pressure. Isoflurane has been
used with great success in guinea pigs. Innovar Vet if injected IM can cause necrosis
and sloughing of the skin at the injection site.

PRIMATES
DRUG
Acepromazine
Malate

ANESTHESIA

ANALGESIA

TRANQUILIZATION

0.5 - 1 mg/kg IM, SC

Aspirin (NSAID)

100 mg/kg PO
q24h

Atropine

0.02-0.05 mg/kg SC,

IM, IV

Buprenorphine

Buprenorphine SR

Butorphanol

Carprofen
(NSAID)

2 mg/kg PO q12h

Diazepam

0.25-0.5 mg/kg IM,

IV

Flunixin (NSAID)

Glycopyrrolate
Ibuprofen
(NSAID)

0.005-0.01 mg/kg IM

10 mg/kg IM q1224h

7 mg/kg PO q12h

Isoflurane
Ketamine
Ketamine +
Acepromazine
Ketamine +
Diazepam
Ketamine +
Dexmedetomidine
Ketamine +

1-3% inhalant to
effect (up to 5% for
induction)
5 - 10 mg/kg IM
K: 4 mg/kg IM
A: 0.04 mg/kg IM
K: 15 mg/kg IM
D: 1 mg/kg IM
K: 3-5 mg/kg IM
D: 0.01-0.03 mg/kg
IM
K: 7 mg/kg IM

0.01 - 0.03 mg/kg

IM q6-12h
0.2 mg/kg SC q5
days
Old World
Primates: 0.5
mg/kg IM q8h
New World
Primates: 0.02
mg/kg SQ q6h

Xylazine

X: 0.5 mg/kg IM

Meloxicam
(NSAID)

Oxymorphone

Propofol
(induction)
Telazol

0.1-0.2 mg/kg PO
q24h for up to 3
days
Old world: 0.15
mg/kg SC, IM
q4-6h
New world: 0.075
mg/kg SC, IM
q4-6h

2.5 - 5.0 mg/kg IV

2 - 6 mg/kg IM

PIG
ANESTHESIA
Premeds

DRUG

ANALGESIA

TRANQUILIZATION

Acepromazine
Aspirin
(NSAID)
Atropine

0.1 - 0.22 mg/kg IM

10 mg/kg PO q12h

0.04 mg/kg IM

Buprenorphine

Butorphanol

0.005 - 0.1 mg/kg IM

q8-12h; use 0.05-0.1
mg/kg for major
surgeries
0.1 - 0.3 mg/kg IM q812h

1 - 2 mg/kg PO q12h

1-3% inhalant to
effect (up to 5% for
induction)

Use Dog Dose

0.5 - 10 mg/kg IM

Carprofen
(NSAID)
Diazepam
Fentanyl
Isoflurane
Ketamine +
Xylazine

K: 12-20 mg/kg IM
X: 1-2 mg/kg IM

Meperidine

Morphine

Propofol
(induction)
Telazol

2 - 10 mg/kg IM q4h
0.2 - 0.9 mg/kg SC, IM
q4h

4-8 mg/kg IV

3-5 mg/kg IM

SHEEP & GOATS

DRUG
Acepromazine

ANESTHESIA

Buprenorphine

Butorphanol

Diazepam
Flunixen
(NSAID)
Isoflurane
Pentobarbital
Propofol
(induction)
Thiopental

Ketamine
Xylazine

Reversal Agents

ANALGESIA

Sheep: 0.015-0.01
mg/kg IM, SQ q612h
Goats: 0.005 mg/kg
IM, SC q6-12h
0.2 1 mg/kg IM, SC
q2-4h

TRANQUILIZATION
0.05 - 0.1 mg/kg IM

Sheep: 1.0 - 1.5 mg/kg IM;

5 mg/kg in feed
Goats: 0.5 - 1.5 mg/kg IM;
15 mg/kg in feed

to effect
20 - 30 mg/kg IV

0.5-1 mg/kg PO, IM,

IV q8hrs

...

2-4 mg/kg IV
Sheep: 10-15
mg/kg IV
Goats:20 - 22
mg/kg IV
K: 15 mg/kg IM,
X:, 0.1-0.2
mg/kg IM

Atipamezole
(Antisedan)

Yohimbine
(Yobine)

Reverses Effects
of
Medetomidine
(Domitor)

Xylazine
(Rompun)

Non-rodent
species
Give IM an equal
vol of Antisedan
as Domitor. May
give IV if it has
been 45 min
since Domitor
was given. May
give at equal
volume or
volume of
Domitor dose.

Rabbits

Other

1 mcg/kg SC,
IV

Mice, Rats,
Gerbils,
Hamsters,
Guinea Pigs

Dogs, Cats,
Primates: 0.100.15 mg/kg IV
slowly.

0.2-1 mg/kg
IM, IV

0.1 0.2 mg/kg

IM, SC

Sheep & Goats:

0.13-0.22 IV
slowly.
Pigs: 0.125-0.3
mg/kg IV slowly.

Local Analgesia for Rodents

0.1-1 mg/kg IM,

SC, IV, IP

Mice, Rats,
Gerbils,
Hamsters,
Guinea Pigs:
0.2-0.5 mg/kg IP

Purpose: To provide additional pain relief directly at the site of surgery before making
the surgical incision (i.e., incisional line block). Local anesthesia can be used in addition
to systemic analgesia such as opioids or NSAIDs to control pain. Local analgesic drugs
should not be used as the sole analgesia for moderate to severe pain.
Route: Local anesthetics are given subcutaneously. Intramuscular and Intravenous
injections must be avoided. Systemic toxicity (seizures, heart rhythm disturbances, and
death) results from over dosage or accidental intravenous injections.
Local Anesthetic
Lidocaine (2%)
(20 mg/ml)

Dose
Dilute to 0.5% (5
mg/ml), do not exceed
7 mg/kg total dose

Without epinephrine
Bupivacaine/Marcaine
(0.5%)

Dilute to 0.25% (2.5

mg/ml), do not exceed
8 mg/kg total dose

Without epinephrine
Ropivacaine/Naropin
(0.2%)

Maximum Volumes:
Weight of Mouse

25 g
35 g
45 g
55 g
Weight of Rat

250 g
350 g
450 g
550 g

No need to dilute, do
not exceed 8 mg/kg
total dose.

Dilutions
1:4 dilution - Take 1
ml of lidocaine (2%)
and add 3 ml of sterile
water. Discard after
using.
1:2 dilution Take 1
ml of 0.5%
bupivacaine and add 1
ml of sterile water.
Discard after using.
N/A

Notes
Faster onset than
bupivacaine but short
(< 1 hr) duration of
action
Slower onset than
lidocaine but longer
(4-8 hr) duration of
action
Slower onset than
lidocaine but longer
(4-8 hr) duration of
action; wider safety
margin

Maximum Volume
Diluted Lidocaine
(0.5%)
Do Not Exceed
0.03 ml
0.05 ml
0.06 ml
0.07 ml

Maximum Volume
Diluted Bupivacaine
(0.25%)
Do Not Exceed
0.08 ml
0.11 ml
0.14 ml
0.17 ml

Maximum Volume
Ropivacaine (0.2%)

Maximum Volume
Diluted Lidocaine
(0.5%)
Do Not Exceed
0.35 ml
0.49 ml
0.63 ml
0.77 ml

Maximum Volume
Diluted Bupivacaine
(0.25%)
Do Not Exceed
0.8 ml
1.12 ml
1.44 ml
1.76 ml

Maximum Volume
Ropivacaine (0.2%)

Use of Hypothermia as Anesthesia for Neonatal Rodents

Do Not Exceed
0.1 ml
0.14 ml
0.18 ml
0.22 ml

Do Not Exceed
1.0 ml
1.4 ml
1.8 ml
2.2 ml

Hypothermia may be used to induce anesthesia in neonatal rats and mice less than six
days of age. Their small size and body mass makes rapid core cooling feasible through
surface cooling. They are also more resistant to arrest of blood supply to the brain and
tolerate extended periods of 1C body temperature without known negative effects.
Potential risks of hypothermia include ventricular fibrillation, tissue hypoxia and
metabolic acidosis on warming.
When a need for hypothermia is demonstrated, the following guidelines should be
followed:
To induce hypothermia, pups may be placed in a latex sleeve or glove and immersed up
to the neck in crushed ice and water (2C-3C) which requires a 5-8 minute induction
time (2-3 minutes to unconsciousness and 3-5 minutes to complete blockage of neural
transmission). Alternatively, pups may be placed in a paper-lined tube and packed in
crushed ice which may require up to 15 minutes to obtain a surgical plane of anesthesia.
Analgesia for hypothermia induced by these methods lasts approximately 10 minutes.
Do not place the animals directly on the cooling medium; provide a cloth, paper, or other
barrier material. Simply placing conscious animals in a cold room or on an ice pack are
not acceptable as induction may take 30-45 minutes.
The anesthetic state may be prolonged by placing the hypothermic pup on an ice pack
(3C-4C). Studies have shown that rodent pups will maintain a core body temperature
of approximately 5C when kept on an ice pack for a maximum of 15 minutes.
Illumination of the surgical field should be fiber optic in nature, because incandescent
bulbs may cause inadvertent and uncontrollable warming.
After the procedure, avoid rapid re-warming because of possible tissue damage. Until
animals are fully conscious they cannot escape' excessive heat generated by
incandescent lamps, heating pads, or other warming devices. Pups can also be placed
in an incubator at 33C for 20-30 minutes. Complete recovery typically requires 30-60
minutes.
Supplemental oxygen may benefit the recovering animals.