The University of Texas at TylerThe University of Texas at TylerInstitutional Animal Care and Use Committee:Institutional Animal Care and Use Committee:

Working with the Laboratory MouseWorking with the Laboratory Mouse

*Note: Content not specific to UT Tyler is from the *Note: Content not specific to UT Tyler is from the American Association for Laboratory Animal Science American Association for Laboratory Animal Science

(AALAS)(AALAS)

Working with the Laboratory Working with the Laboratory MouseMouse

The goal of this course is to provide information on the The goal of this course is to provide information on the use of mice in research. This course will address:use of mice in research. This course will address:

Regulatory requirements Regulatory requirements Occupational Health and Safety Occupational Health and Safety Housing and Routine Care Housing and Routine Care Animal Identification Animal Identification Sex Determination Sex Determination Clinical Signs of Illness Clinical Signs of Illness Humane Endpoints Humane Endpoints EuthanasiaEuthanasia

Regulatory Requirements Regulatory Requirements

To ensure the humane treatment of laboratory To ensure the humane treatment of laboratory animals, animal research is regulated by two animals, animal research is regulated by two federal agencies:federal agencies:• The United States Department of Agriculture (The United States Department of Agriculture (USDAUSDA) / ) /

Animal Care enforcing the Animal Welfare Act Animal Care enforcing the Animal Welfare Act RegulationsRegulations

• The National Institutes of Health / Office of Laboratory The National Institutes of Health / Office of Laboratory Animal Welfare (Animal Welfare (OLAWOLAW) enforcing the PHS Policy) enforcing the PHS Policy

The Animal Welfare Act Regulations and the PHS The Animal Welfare Act Regulations and the PHS Policy mandates differ greatly with respect to the Policy mandates differ greatly with respect to the laboratory strains of mice and rats. These species laboratory strains of mice and rats. These species are not covered by the USDA but are included in are not covered by the USDA but are included in PHS regulations and policy. PHS regulations and policy.

Regulatory Requirements Regulatory Requirements

The UT Tyler The UT Tyler IACUCIACUC has policies and has policies and procedures on the use of animals that reflect procedures on the use of animals that reflect federal and other government mandates and federal and other government mandates and professional standards in the laboratory animal professional standards in the laboratory animal field—refer to the UT Tyler IACUC Policy field—refer to the UT Tyler IACUC Policy Handbook available at:Handbook available at:

http://www.uttyler.edu/research/animals.htmhttp://www.uttyler.edu/research/animals.htm

Occupational Health & Safety Occupational Health & Safety Public Health Service Policy Public Health Service Policy

• The Public Health Service Policy requires The Public Health Service Policy requires institutions to have an institutions to have an occupational health and safety program for for individuals working with laboratory animals. individuals working with laboratory animals. This requirement is also reiterated in the This requirement is also reiterated in the Guide. .

• It is the responsibility of principal It is the responsibility of principal investigators to ensure that their laboratory investigators to ensure that their laboratory staff are informed of and participate in UT staff are informed of and participate in UT Tyler’s occupational health and safety Tyler’s occupational health and safety program.program.

Occupational Health and Safety: Occupational Health and Safety: Hazards of Working with Mice Hazards of Working with Mice

Working with mice is associated with Working with mice is associated with the following hazards which will be the following hazards which will be described more fully in the following described more fully in the following pages:pages:

• InjuriesInjuries• AllergiesAllergies• ZoonosesZoonoses

Occupational Health & Safety Occupational Health & Safety Injuries and Allergies Injuries and Allergies

InjuriesInjuries• Personnel handling a mouse can be bitten if the animal is Personnel handling a mouse can be bitten if the animal is

poorly restrained. Though mice are often inclined to bite poorly restrained. Though mice are often inclined to bite when frightened, their incisors do not always break the when frightened, their incisors do not always break the skin.skin.

• Poor technique in handling and restraining a mouse may Poor technique in handling and restraining a mouse may also cause injury to the mouse.also cause injury to the mouse.

• Training staff to work effectively and humanely with mice is Training staff to work effectively and humanely with mice is essential to prevent injuries to people and mice.essential to prevent injuries to people and mice.

AllergiesAllergies• People can develop an allergy to rodents after having People can develop an allergy to rodents after having

contact with them for some time.contact with them for some time.• Persons who develop allergy symptoms should seek Persons who develop allergy symptoms should seek

medical counseling and may have to discontinue working medical counseling and may have to discontinue working with this species.with this species.

Housing and Routine Care:Housing and Routine Care:Animal CrowdingAnimal Crowding

Animal crowding in a cage affects environmental Animal crowding in a cage affects environmental quality (the accumulation of urine, for example, quality (the accumulation of urine, for example, leads to excess ammonia and moisture).leads to excess ammonia and moisture).

Crowding can also cause newborn pups to be injured Crowding can also cause newborn pups to be injured or killed. Crowding is a special concern for multiple or killed. Crowding is a special concern for multiple litters in a cage since pups grow very quickly and litters in a cage since pups grow very quickly and rapidly increase their output of excreta.rapidly increase their output of excreta.

In particular, if the mother is about to give birth to a In particular, if the mother is about to give birth to a second litter, the first litter should be weaned and second litter, the first litter should be weaned and removed to new cages to prevent smothering and removed to new cages to prevent smothering and trauma of the newborns.trauma of the newborns.

Routine Sanitation Routine Sanitation

Routine sanitation and environmental controls are necessary Routine sanitation and environmental controls are necessary for protecting animal health and for minimizing the for protecting animal health and for minimizing the introduction for non-experimental variables which could introduction for non-experimental variables which could undermine the quality of research data.undermine the quality of research data.

Sanitation schedules vary according to the type of mouse Sanitation schedules vary according to the type of mouse caging.caging.

Based on the types of caging and bedding in use at your Based on the types of caging and bedding in use at your facility, your institution will have a standard operating facility, your institution will have a standard operating procedure (procedure (SOPSOP) on the sanitation schedule for mouse ) on the sanitation schedule for mouse cages.cages.

Environmental Controls Environmental Controls The The Guide for the Care and Use of Laboratory AnimalsGuide for the Care and Use of Laboratory Animals establishes establishes

standards for laboratory animal environments in regard to: for laboratory animal environments in regard to:

• room temperature room temperature • humidity humidity • ventilation ventilation • illumination and light schedule illumination and light schedule • noise abatementnoise abatement• The details of these standards are generally not a concern to The details of these standards are generally not a concern to

researchers who house their animals in an institutional animal researchers who house their animals in an institutional animal facility, because it is the animal care staff who have the facility, because it is the animal care staff who have the responsibility of operating the facility in compliance with these responsibility of operating the facility in compliance with these environmental standards.environmental standards.

• For practical considerations due to common work hours, researchers For practical considerations due to common work hours, researchers should be aware of the lighting schedules used in the rodent should be aware of the lighting schedules used in the rodent housing rooms (commonly 12 hr light:12 hr dark or 14 hr light:10 hr housing rooms (commonly 12 hr light:12 hr dark or 14 hr light:10 hr dark).dark).

• If researchers working late turn on the lights in the animal rooms If researchers working late turn on the lights in the animal rooms during the dark period, the disruption of the light schedule may during the dark period, the disruption of the light schedule may cause the animals to be perturbed, which may have effects on cause the animals to be perturbed, which may have effects on breeding performance and on circadian rhythms.breeding performance and on circadian rhythms.

Animal Handling and Restraint Animal Handling and Restraint

It's important to remember the following:It's important to remember the following:

• As a small animal, mice can be easily injured if As a small animal, mice can be easily injured if handled roughly. You should learn how to handled roughly. You should learn how to handle them firmly but gently and with handle them firmly but gently and with confidence to avoid injuring these delicate confidence to avoid injuring these delicate animals.animals.

• Mice are inclined to become aggressive and Mice are inclined to become aggressive and bite. Although their teeth seldom break bite. Although their teeth seldom break through your skin, a bite can hurt! Develop through your skin, a bite can hurt! Develop your confidence in handling mice by learning your confidence in handling mice by learning from an experienced mentor and practice hand from an experienced mentor and practice hand restraint first on anesthetized mice.restraint first on anesthetized mice.

Animal Handling and Restraint Animal Handling and Restraint

If a mouse does bite you, train yourself not to If a mouse does bite you, train yourself not to fling the animal from your hand as a fall can hurt fling the animal from your hand as a fall can hurt the animal and cause the loss of an animal and the animal and cause the loss of an animal and possible future genetic contamination of strains possible future genetic contamination of strains and stocks. and stocks.

Instead, remember that the mouse has bitten you Instead, remember that the mouse has bitten you in fear and self-defense. At that moment, you and in fear and self-defense. At that moment, you and the mouse share a mutual desire to disengage the mouse share a mutual desire to disengage from each other. from each other.

The best way to remove a mouse that is hanging The best way to remove a mouse that is hanging on to your finger is to train yourself to lower your on to your finger is to train yourself to lower your hand back into its cage. Presented with a retreat hand back into its cage. Presented with a retreat to its home cage, the mouse will quickly jump off to its home cage, the mouse will quickly jump off your finger.your finger.

Animal Handling and Restraint Animal Handling and Restraint

The succeeding screens will show: The succeeding screens will show:

• Common approaches for hand restraint of Common approaches for hand restraint of mice.mice.

• Two devices commonly used to restrain Two devices commonly used to restrain mice.mice.

Hand Restraint Hand Restraint There are two common hand methods for There are two common hand methods for

restraining mice. No matter how you will restrain restraining mice. No matter how you will restrain the mouse, mice are picked up the following way:the mouse, mice are picked up the following way:

• If your mice are in barrier or containment housing, If your mice are in barrier or containment housing, please open their cages in an appropriate hood or please open their cages in an appropriate hood or laminar-flow workbench.laminar-flow workbench.

• Remove the cage top if they are housed in a filter-top Remove the cage top if they are housed in a filter-top cage.cage.

• Place the wirelid top sideways on top of the cage.Place the wirelid top sideways on top of the cage.• Pick up a mouse by the tail (away from the tail tip) and Pick up a mouse by the tail (away from the tail tip) and

lift the mouse directly to the wirelid. You will find that lift the mouse directly to the wirelid. You will find that the wirelid is a useful area to which the mouse will want the wirelid is a useful area to which the mouse will want to hang on with its front feet, allowing you the to hang on with its front feet, allowing you the opportunity to reposition your grasp.opportunity to reposition your grasp.

Tail Wrap MethodTail Wrap Method

In the In the tail wraptail wrap method, you transfer the tail method, you transfer the tail from your thumb and forefinger to the 4th and from your thumb and forefinger to the 4th and 5th fingers of the same hand. These fingers press 5th fingers of the same hand. These fingers press into your palm the mouse's tail, close to the into your palm the mouse's tail, close to the animal's rump. Then, your thumb and forefinger animal's rump. Then, your thumb and forefinger grasp a scruff of skin at the occiput of the mouse. grasp a scruff of skin at the occiput of the mouse. In this method, the mouse is held by two points: In this method, the mouse is held by two points: the occiput and the base of the tail. To further the occiput and the base of the tail. To further immobilize the hindquarters, you may gently immobilize the hindquarters, you may gently entrap one leg along with the tail beneath your entrap one leg along with the tail beneath your 5th finger. It is important to provide back support 5th finger. It is important to provide back support to the mouse and avoid hyperextension of its to the mouse and avoid hyperextension of its back. back.

Points to remember:Points to remember: When picking up a mouse –When picking up a mouse –

• Pick up the tail at the middle, not the tip. A mouse does not Pick up the tail at the middle, not the tip. A mouse does not need to be picked up at the base of the tail like a rat does, need to be picked up at the base of the tail like a rat does, because the mouse is light and its weight will not damage because the mouse is light and its weight will not damage its tail.its tail.

• Never dangle a mouse by its tail, but instead lift it directly Never dangle a mouse by its tail, but instead lift it directly to the cage wirelid or some other support. When dangled, to the cage wirelid or some other support. When dangled, mice appear anxious as seen by the their behavior in mice appear anxious as seen by the their behavior in hyperextending their feet.hyperextending their feet.

• Mice feel more secure when given a protective cover, e.g., Mice feel more secure when given a protective cover, e.g., by an overturned cup-like device. This is a means to hold a by an overturned cup-like device. This is a means to hold a mouse securely and comfortably for a few moments.mouse securely and comfortably for a few moments.

• If you need a place to briefly sort and hold your mice, say If you need a place to briefly sort and hold your mice, say while you are rapidly administering injections to a cage of while you are rapidly administering injections to a cage of mice, each mouse can be placed on the wirelid after its mice, each mouse can be placed on the wirelid after its injection. Mice will stay on their wirelid a short while if food injection. Mice will stay on their wirelid a short while if food blocks are present, due to their instincts for food. If you blocks are present, due to their instincts for food. If you quickly make all your injections, all the mice can be treated quickly make all your injections, all the mice can be treated without a mixup of repeated or skipped administrations.without a mixup of repeated or skipped administrations.

Determining Sex and AgeDetermining Sex and Age Refer to the image below: the top two mice are neonates and note that the Refer to the image below: the top two mice are neonates and note that the

anogenital distance is larger in the male than in the female neonates, the anogenital distance is larger in the male than in the female neonates, the penis and vulva cannot be easily differentiated and so are referred to as a penis and vulva cannot be easily differentiated and so are referred to as a genital papillagenital papilla. The bottom two animals are adults; genitalia are . The bottom two animals are adults; genitalia are differentiated.differentiated.

Determining Sex and AgeDetermining Sex and Age

• Also, nipples become evident in females at about 10 days Also, nipples become evident in females at about 10 days of age.of age.

• Male mice, like other rodent species, retain an open Male mice, like other rodent species, retain an open inguinal canal in adulthood. That is, the descended testicles inguinal canal in adulthood. That is, the descended testicles communicate with the abdominal cavity. Depending on the communicate with the abdominal cavity. Depending on the position in which the mouse is held, the testicles may be position in which the mouse is held, the testicles may be retracted into the abdomen or descended into the scrotum. retracted into the abdomen or descended into the scrotum.

• Because of the open inguinal canal, castration of mice Because of the open inguinal canal, castration of mice requires that the surgeon use caution when applying requires that the surgeon use caution when applying tension to the testicle. Too much tension can result in the tension to the testicle. Too much tension can result in the intestines being pulled through the inguinal canal.intestines being pulled through the inguinal canal.

Determining Sex and AgeDetermining Sex and Age

• Occasionally people experience difficulty in determining Occasionally people experience difficulty in determining gender in mice at weaning age, although the anogenital gender in mice at weaning age, although the anogenital distances are markedly different between males and distances are markedly different between males and females.females.

• Newborns have a subtle difference in the anogenital Newborns have a subtle difference in the anogenital distance, due to their small size and scale of the anatomical distance, due to their small size and scale of the anatomical landmarks. In determining the gender of newborns, it’s best landmarks. In determining the gender of newborns, it’s best to examine several animals side by side to distinguish the to examine several animals side by side to distinguish the males from the females.males from the females.

• The difference becomes more apparent after a few days of The difference becomes more apparent after a few days of age. Another landmark is the presence of nipples in the age. Another landmark is the presence of nipples in the females from 10 days of age, which are absent in the male. females from 10 days of age, which are absent in the male. Darker mice are more difficult to differentiate than light Darker mice are more difficult to differentiate than light colored mice.colored mice.

Sex and AgeSex and Age

Sex and AgeSex and Age

Newborn through Day 5 Newborn through Day 5

The images on the next slide show pups from The images on the next slide show pups from 1 to 5 days of age. (The following slides 1 to 5 days of age. (The following slides show mouse pups from 6 to 15 days of show mouse pups from 6 to 15 days of age.)age.)

Newborn, or 1 day old, mice are very red, Newborn, or 1 day old, mice are very red, helpless, and hairless. Because of their helpless, and hairless. Because of their color and lack of hair, they are often color and lack of hair, they are often referred to as referred to as pinkiespinkies..

Sex and AgeSex and Age

Sex and AgeSex and Age

Day 6 through 10 Day 6 through 10

The next slide shows images of pups at The next slide shows images of pups at ages of 6 through 10 days. The ages of 6 through 10 days. The stages of fur growth are an important stages of fur growth are an important indicator of age during this period of indicator of age during this period of timetime

Sex and AgeSex and Age

Here are images of Here are images of pups at ages of 6 pups at ages of 6 through 10 days. The through 10 days. The stages of fur growth stages of fur growth are an important are an important indicator of age during indicator of age during this period of time.this period of time.

Sex and AgeSex and Age

Day 11 through 28 Day 11 through 28

The next slide shows images of pups The next slide shows images of pups from 11 days through 4 weeks of from 11 days through 4 weeks of age. age.

This 18 day period is one of rapid This 18 day period is one of rapid growth and change as their eyes growth and change as their eyes open and activity increases.open and activity increases.

Sex and AgeSex and Age

Reproductive Data Of MiceReproductive Data Of Mice

StageStage• Sexual maturitySexual maturity• Estrous cycleEstrous cycle• GestationGestation• Weaning ageWeaning age• Reproductive lifeReproductive life• Life spanLife span

• Pups begin eatingPups begin eating

dry fooddry food

AgeAge• 40-60 days40-60 days• 4 – 5 days4 – 5 days• 21 days21 days• 21-30 days21-30 days• ±8 months±8 months• 1 - 3 years1 - 3 years• 12 – 14 days12 – 14 days

Minimizing Pain and Distress Minimizing Pain and Distress WHEREVER POSSIBLE, PAIN/DISTRESS SHOULD BE WHEREVER POSSIBLE, PAIN/DISTRESS SHOULD BE

ELIMINATEDELIMINATED!!

Clinical Signs of IllnessClinical Signs of Illness

• Achieving humaneness in animal research depends upon the Achieving humaneness in animal research depends upon the control and, whenever possible, the reduction of animal pain and control and, whenever possible, the reduction of animal pain and distress. distress.

• Minimizing pain and distress also reduces the impact of these Minimizing pain and distress also reduces the impact of these extraneous factors on the research, i.e., as sources of non-extraneous factors on the research, i.e., as sources of non-experimental variation.experimental variation.

For example, in a mouse model of experimental autoimmune For example, in a mouse model of experimental autoimmune encephalomyelitis, implementation of supportive treatment encephalomyelitis, implementation of supportive treatment (hydration and nutrition) was shown to protect against loss of body (hydration and nutrition) was shown to protect against loss of body weight and to greatly extend survival of the animals in the study, weight and to greatly extend survival of the animals in the study, from 25 to 60 days. (Ref.: from 25 to 60 days. (Ref.: Lab AnimalLab Animal, 29(5): 40-46, 2000.) , 29(5): 40-46, 2000.)

Minimizing Pain and DistressMinimizing Pain and DistressCauses of Pain and Distress in MiceCauses of Pain and Distress in Mice

• Research staff should be familiar with the causes Research staff should be familiar with the causes of animal pain and distress. Pain and distress are of animal pain and distress. Pain and distress are caused by spontaneous and experimentally-caused by spontaneous and experimentally-induced disease or injury.induced disease or injury.

• Many factors may contribute to an animal’s Many factors may contribute to an animal’s distress or discomfort, including extreme distress or discomfort, including extreme homeostatic challenges.homeostatic challenges.

• Pain/distress should be minimized to an extent Pain/distress should be minimized to an extent that is possible and compatible with experimental that is possible and compatible with experimental objectives.objectives.

Factors in Pain & Distress Factors in Pain & Distress Changes in the following parameters may cause or be associated with animal Changes in the following parameters may cause or be associated with animal

pain or distress:pain or distress:

temperature (environmental and body temperature)temperature (environmental and body temperature) hypoxiahypoxia edemaedema blood electrolytes, e.g. hyperkalemiablood electrolytes, e.g. hyperkalemia dehydrationdehydration environment environment

• caging caging • cage mates cage mates • lighting lighting • humidity humidity • noise noise • VibrationVibration

Note - Smaller mammals experience physiologic changes such as starvation Note - Smaller mammals experience physiologic changes such as starvation (due to high metabolic rate) and chilling (due to large ratio of body surface (due to high metabolic rate) and chilling (due to large ratio of body surface area to mass) faster than larger animals.area to mass) faster than larger animals.

Signs of Pain and DistressSigns of Pain and Distress• Signs of pain and distress in rodents are not easy to Signs of pain and distress in rodents are not easy to

detect because of their body size, their tendency to detect because of their body size, their tendency to conceal outward signs of pain and distress, and their conceal outward signs of pain and distress, and their habit of hiding or freezing when disturbed.habit of hiding or freezing when disturbed.

• Nevertheless, signs of pain or distress can be detected Nevertheless, signs of pain or distress can be detected in rodents by carefully observing subtle changes in in rodents by carefully observing subtle changes in behavior.behavior.

• The ability to properly assess pain and distress in The ability to properly assess pain and distress in rodents requires:rodents requires:

--knowledge of normal rodent behavior and --knowledge of normal rodent behavior and appearance appearance --a systematic approach to observing clinical signs in --a systematic approach to observing clinical signs in rodentsrodents

Assessing Appearance and BehaviorAssessing Appearance and Behavior

From the Cage ExteriorFrom the Cage Exterior • Routinely inspect the rodents through the top and sides of Routinely inspect the rodents through the top and sides of

the cage. Get in the habit of removing the cage from the the cage. Get in the habit of removing the cage from the shelf and looking through all sides of the cage. Signs of shelf and looking through all sides of the cage. Signs of distress may be missed in animals on lower or upper distress may be missed in animals on lower or upper shelves because of low lighting or difficult access.shelves because of low lighting or difficult access.

• Baby mice and rats can be inconspicuous within piles of Baby mice and rats can be inconspicuous within piles of bedding or nestboxes.bedding or nestboxes.

Cage Wirelid OffCage Wirelid Off• Lift the cage wirelid to elicit a response to your presence. Lift the cage wirelid to elicit a response to your presence.

This disturbance may prompt the animals to move about This disturbance may prompt the animals to move about the cage. Examine the animals’ behavior, gait, and hair the cage. Examine the animals’ behavior, gait, and hair coat.coat.

• Normal mice are inquisitive and explore their cage Normal mice are inquisitive and explore their cage perimeter.perimeter.

Behaviors to ObserveBehaviors to Observe• Abnormal mice may huddle in their cage, or they may fail to move Abnormal mice may huddle in their cage, or they may fail to move

around and explore their cage. In addition, mice may vocalize when around and explore their cage. In addition, mice may vocalize when approached.approached.

• Inspect the animal’s mode and speed of movement. Observe the tail Inspect the animal’s mode and speed of movement. Observe the tail position when the animal moves.position when the animal moves.

• Is the gait (how it walks) awkward? Observe how all limbs move Is the gait (how it walks) awkward? Observe how all limbs move while walking. while walking.

• Does the animal teeter or stumble? Does the animal teeter or stumble? • Is the animal’s back hunched and abdomen tucked while walking? Is the animal’s back hunched and abdomen tucked while walking? • Is the tail held stiff and upright? Or does the tail drag?Is the tail held stiff and upright? Or does the tail drag?

Tip:Tip: Observe a cage of normal animals for a comparison. Observe a cage of normal animals for a comparison.

• Examine (and treat) an individual mouse by gently restraining the Examine (and treat) an individual mouse by gently restraining the animal. You can move the animal to a separate examination box for animal. You can move the animal to a separate examination box for detailed clinical inspectiondetailed clinical inspection..

Red TearsRed Tears

Stressed mice and rats commonly display Stressed mice and rats commonly display red tearsred tears or or porphyrin stainingporphyrin staining. Porphyrin is an oily discharge from . Porphyrin is an oily discharge from the Harderian glands located in the orbit behind the the Harderian glands located in the orbit behind the eyes.eyes.

Porphyrin staining may be seen:Porphyrin staining may be seen:• on the nose, on the nose, • around the eyelids, around the eyelids, • or on the medial aspect of the forepaws that become or on the medial aspect of the forepaws that become

stained through grooming of the face.stained through grooming of the face.

Affected rodents may also fail to groom or they may have Affected rodents may also fail to groom or they may have piloerection of the hair coat (giving a spiky appearance piloerection of the hair coat (giving a spiky appearance to the hair).to the hair).

Types of Observations Types of Observations A common approach to assessing animal appearance and behavior is A common approach to assessing animal appearance and behavior is

through observation of the following:through observation of the following:

Activity Level Activity Level • e.g., hypoactivity (hunched, huddled, lethargic), hyperactivity, e.g., hypoactivity (hunched, huddled, lethargic), hyperactivity,

restlessness, lack of inquisitiveness  restlessness, lack of inquisitiveness 

Attitude Attitude • e.g., arousal, depression, awareness of surroundings  e.g., arousal, depression, awareness of surroundings 

Behavior, Spontaneous Behavior, Spontaneous • e.g., vocalization, self-trauma, isolation from cage mates. These e.g., vocalization, self-trauma, isolation from cage mates. These

observations are made without disturbing the animal.  observations are made without disturbing the animal. 

Behavior, Provoked Behavior, Provoked • e.g., vocalization, hiding, aggressiveness, minimal response. These e.g., vocalization, hiding, aggressiveness, minimal response. These

observations are made when the animal is disturbed or even observations are made when the animal is disturbed or even prodded. prodded. 

Types of ObservationsTypes of Observations Body Condition Body Condition

• e.g., emaciation, missing anatomy  e.g., emaciation, missing anatomy 

Food and Fluid Intake Food and Fluid Intake • e.g., elimination of feces and urine  e.g., elimination of feces and urine 

Fur and Skin Fur and Skin • e.g., unkempt or greasy or dull fur; porphyrin staining around eyes e.g., unkempt or greasy or dull fur; porphyrin staining around eyes

and nostrils; cyanotic, pale, or congested mucous membranes or and nostrils; cyanotic, pale, or congested mucous membranes or skin (ears, feet, tail); skin lesions; soiled anogenital area  skin (ears, feet, tail); skin lesions; soiled anogenital area 

Eyes Eyes • e.g., clarity/condition of lens, cornea; position of globe (e.g., e.g., clarity/condition of lens, cornea; position of globe (e.g.,

sunken in orbit or protruding); condition of eyelids, encrustation  sunken in orbit or protruding); condition of eyelids, encrustation 

Posture Posture • e.g., hunched back, tucked abdomen; prostrate; head tucked e.g., hunched back, tucked abdomen; prostrate; head tucked

down down 

Types of ObservationsTypes of Observations Locomotion Locomotion

• e.g., gait, ataxia, lameness, action of each limb, position of tail e.g., gait, ataxia, lameness, action of each limb, position of tail when ambulating  when ambulating 

Neurological Neurological • e.g., tremor, convulsion, circling, paralysis, head tilt, coma  e.g., tremor, convulsion, circling, paralysis, head tilt, coma 

Vital Signs Vital Signs • e.g., respiratory distress (open mouth breathing, pronounced e.g., respiratory distress (open mouth breathing, pronounced

chest movement)  chest movement) 

Other clinical parameters that are relevant to your study Other clinical parameters that are relevant to your study • e.g., presence and status of tumors, infection, or surgical e.g., presence and status of tumors, infection, or surgical

woundswounds

Tip:Tip: It is helpful to have blank forms to use as "score sheets" to It is helpful to have blank forms to use as "score sheets" to enter and track each parameter assessed.enter and track each parameter assessed.

Physical Examination Physical Examination • After assessing the animals’ appearance and behavior, conduct a After assessing the animals’ appearance and behavior, conduct a

physical exam using methods that are appropriate to the species physical exam using methods that are appropriate to the species and experimental model.and experimental model.

• Performing a clinical exam on rodents is somewhat limited Performing a clinical exam on rodents is somewhat limited compared to larger animals due to the greater difficulty in venous compared to larger animals due to the greater difficulty in venous access and the smaller sampling size of biological fluids. access and the smaller sampling size of biological fluids. Nevertheless, specific methods and equipment for rodents allow a Nevertheless, specific methods and equipment for rodents allow a clinical exam to provide information on animal well being.clinical exam to provide information on animal well being.

• Use quantifiable characteristics whenever possible. These can be Use quantifiable characteristics whenever possible. These can be tracked over time and compared to a starting baseline or to normal, tracked over time and compared to a starting baseline or to normal, untreated animals.untreated animals.

• Such measurements are not only helpful for clinical assessments, Such measurements are not only helpful for clinical assessments, but they can also be useful when compiling research data and but they can also be useful when compiling research data and writing manuscripts. Later in this course, simple record-keeping writing manuscripts. Later in this course, simple record-keeping methods will be discussed to help utilize this information.methods will be discussed to help utilize this information.

Physical ExaminationPhysical Examination

You may evaluate:You may evaluate:• BehaviorBehavior• Body weightBody weight• Surface lesions (wounds, masses)Surface lesions (wounds, masses)• Hydration statusHydration status• Body temperature (telemetric methods)Body temperature (telemetric methods)• Blood parameters (Blood collection can be difficult/stressful Blood parameters (Blood collection can be difficult/stressful

in mice; may be used to confirm disease or failed in mice; may be used to confirm disease or failed treatment.)treatment.)

• For specific recommendations on evaluating these For specific recommendations on evaluating these parameters and supportive procedures, refer to the AALAS parameters and supportive procedures, refer to the AALAS Learning Library course Learning Library course Post-Procedure Care of Mice and Post-Procedure Care of Mice and Rats in Research: Minimizing Pain and DistressRats in Research: Minimizing Pain and Distress..

Humane Endpoints Humane Endpoints

Defining Defining ProcedureProcedure

• All of the federal laws, regulations, policies and All of the federal laws, regulations, policies and guidelines applicable to animal research have a guidelines applicable to animal research have a core requirement for the humane treatment of core requirement for the humane treatment of the animals involved in a study.the animals involved in a study.

• Accordingly, your Accordingly, your IACUCIACUC will have requirements will have requirements for the proper care (peri-procedural care) of your for the proper care (peri-procedural care) of your animals prior to, during, and after a research animals prior to, during, and after a research procedure.procedure.

• What is a procedure? A procedure is any activity What is a procedure? A procedure is any activity carried out on the animal, such as a behavioral carried out on the animal, such as a behavioral observation, a venipuncture, or a surgery.observation, a venipuncture, or a surgery.

Humane EndpointsHumane Endpoints

Defining Defining ProcedureProcedure (cont) (cont)

• This peri-procedural care requirement This peri-procedural care requirement covers:covers:

• properly preparing the animal to humanely properly preparing the animal to humanely undergo the procedure,undergo the procedure,

• supporting the animal’s physiological supporting the animal’s physiological function during the procedure, andfunction during the procedure, and

• providing additional supportive care to aid providing additional supportive care to aid the animal in recovering from the the animal in recovering from the procedure.procedure.

Humane EndpointsHumane EndpointsResponsibility for CareResponsibility for Care

• The investigator has the responsibility to see that staff The investigator has the responsibility to see that staff working with the animals are properly trained to not only working with the animals are properly trained to not only perform the procedure humanely but also to provide the perform the procedure humanely but also to provide the necessary supportive care to the animals.necessary supportive care to the animals.

• When performing any procedure, such as a blood collection, When performing any procedure, such as a blood collection, you should think through the steps that are necessary to you should think through the steps that are necessary to protect the animal’s welfare.protect the animal’s welfare.

• For example, for blood collection, you should limit the For example, for blood collection, you should limit the volume taken acutely or chronically. And, with any volume taken acutely or chronically. And, with any venipuncture, you should be prepared to care for the venipuncture, you should be prepared to care for the animal in the event of trauma to the vein area or excess animal in the event of trauma to the vein area or excess hemorrhage. Refer to your institution’s IACUC and hemorrhage. Refer to your institution’s IACUC and attending veterinarian for specific guidelines.attending veterinarian for specific guidelines.

Acclimation Acclimation

• Upon arrival to your facility, your mice should have an Upon arrival to your facility, your mice should have an acclimation periodacclimation period before they are used in research before they are used in research studies. studies.

• This period of time allows animals to adapt to a new This period of time allows animals to adapt to a new environmentenvironment. .

• Effects of transportation stress include alterations in various Effects of transportation stress include alterations in various blood parameters, immune cell function, food intake, and blood parameters, immune cell function, food intake, and animal behavior. animal behavior.

• The period of time necessary for biological stabilization will The period of time necessary for biological stabilization will depend on the parameters to be studied. depend on the parameters to be studied.

• Refer to your institution's attending veterinarian for Refer to your institution's attending veterinarian for recommendations that are appropriate for your project. recommendations that are appropriate for your project. Typically, acclimation periods can range from days to over Typically, acclimation periods can range from days to over a week, depending on the studies involved.a week, depending on the studies involved.