LABORATORY ANIMAL MANAGEMENT AND 3.

Serve on Animal Care and Use Committees

PRODUCTION (IACUC)
4. Provide diagnostic services
5. Provide advice on regulatory issues
6. Conduct research
LECTURE 1: INTRODUCTION

Animal Experimentation - aka animal testing; Animal Welfare - the physical and mental state of
procedures performed on live animals (e.g., mice, an animal in relation to the conditions in which it
rats, rabbits, guinea pig, etc.) for research lives and dies (Definition by OIE)
purposes.
FIVE FREEDOMS
HISTORICAL OVERVIEW 1. Freedom from hunger or thirst
384-322 B.C - studies made by Aristotle which 2. Freedom from discomfort
involved animals. 3. Freedom from pain, injury, or disease
129-199 A.D. - Galen was the first one to conduct 4. Freedom to express (most) normal
demonstration with live animal experimentation; behavior
vivisection 5. Freedom from fear and distress
Greece - records of animal experimentation 2000
years ago THE HARMS TO ANIMALS IN RESEARCH
12th century lbn Zuhr (Avenzoar) - experimented 1. Stress - refers to body's adaptive and
on animals to test surgical procedures before coping - responding process when the
applying to human patients individuals face or perceive threats or
1628 - William Harvey published his work on heart challenges to the body caused by
and the movement of blood in animals environmental charges.
1800s - France as one of the leading centers of Stress Input - aka stressors; refers to
experimental biology and medicine environmental or stimuli that may
Francois Magendie - experimental physiology cause physiological reactions after the
Claude Bernard - experimental medicine body performs cognition evaluation
Louis Pasteur - microbiology and immunology - Internal environment - production
Laboratory Animals - any vertebrate animal of all the necessary materials and
produced or used for research, testing, and balance disorders within the body
academic purposes - External environment -
environmental conditions of the
The THREE R's body
 Replacement - Psycho-social environment - living
 Refinement conditions
 Reduction 2. Pain - unpleasant feeling caused by
emotional perceptions along with existing
IACUC - Institutional Animal Care and Use potential body injury
Committee 3. Distress and Discomfort - occurs when an
animal is unable to adapt completely to a
ADVANTAGES OF USING LABORATORY ANIMALS stressor
 Easy handling
 Ease of obtaining samples MONITORING THE CARE AND USE OF ANIMALS
 Adequate number of animals
 Short breeding period IACUC - Institutional Animal Care and Use
 High number of pups Committee (IACUC) – oversee and evaluate the
 Easy colony maintenance institution’s animal program, procedures, and
 Low cost for operation and maintenance facilitates to ensure that they are consistent with
 Published and established protocols the recommendations in this guide.

Animal care and use protocols - various topics

LECTURE 2: WELFARE OF LABORATORY ANIMALS such as rationale or purpose of the proposed use
of animals are considered in the preparation and
Biomedical Research - broad area of science that
review of animal care and use protocols.
looks for ways to prevent and treat diseases that
cause illness and death in people and in animals
Physical restraint - the use of manual or
mechanical means to limit some or all of an
DUTIES OF LABORATORY ANIMAL
animal’s normal movement for several purposes
VETERINARIANS
such as veterinary examination
1. Provide veterinary care and expertise
relating to husbandry and welfare
2. Monitor disease control programs
Multiple major surgical procedures - penetrates housed in tightly controlled and monitored
and exposes a body cavity or produces substantial in isolation to prevent contamination
impairment of physical or physiologic function. 2. Axenic animals - also germ-free mice but
Food or fluid restriction - if required, at least are purposively induced later with 1 or
minimal quantities of food and fluid should be more known non-pathogenic microflora
available to provide for development of young 3. Defined microbially associated animals -
animals and to maintain long-term well-being of all limited known non-pathogenic
animals microorganism
4. Barrier-maintained animals - mice
Veterinary care - adequate veterinary care must maintained in a barrier unit or barrier
be provided, including access to all animals for cabinets
evaluation of their health and well-being 5. Monitored animals - mice bred and raised
in conventional room and filter-capped
Personnel qualifications & training - the policy cages
requires institutions to ensure that people caring 6. Conventional animals - mice that are
for or using animals are qualified to do. handled with unknown microbes

Occupational health & safety to personnel - a GENERAL BIOLOGICAL CHARACTERISTICS

safety program for this must be consistent with the  Body is covered with hair
state and local regulations and should focus on  Face is cuspate with whiskers on both
maintaining a safe and healthy workplace sides
 Ears are semicircular
 Bid bright red eyes
LECTURE 3: MICE  Body length - 10-15 cm (adult mouse);
Mice almost same length with the tail
 Genu: Mus  Tail is covered with short hairs and horny
 Subfamily: Murinae scales
 Family: Muridae  Different hair colors (e.g., white, gray,
 Order: Rodentia black, brown, yellow, chocolate,
 Laboratory strains are derived from: cinnamon)
Mus musculus musculus (from Eastern  Absence of sweat glands
Europe) and Mus musculus domesticus  Poor adaptation the environment
(Western Europe)  Docile and easy to handle
 474 inbred strains, over 200 outbred  Likes to live in dimly lit place
strains and more than 2000 mutant strains  Nocturnal
 Most commonly used laboratory mammal  Feeding, mating and parturition occurs at
night
 Most active during the predawn hours and
Example of Laboratory MICE 1-2 hrs after evening
 Social animals; grow faster when fed in
groups (overcrowding might inhibit
reproductive capacity)
 Barbering – hierarchal system where
dominant mice chew off the hair of mice
on lower ranks
 Early sexual maturity
 Male mice will frequently bite others that
they have not fathered
 Whiskers are retained in dominant mice;
subordinate mice lack hair and whiskers
 Extremely sensitive to stimuli (e.g. light,
noise, and odors); overexposure can lead
COMMON LABORATORY MICE IN THE
to cannibalism
PHILIPPINES

1. ICR mice (outbred strain)

2. BALB/c mice (inbred strain)

ECOLOGICAL CLASSES OF MICE

1. Gnotobiotic animals - mice that lack all
microorganisms; microbiologically sterile;
 Male: paired testes, urethra, penis and
associated duct and glands (e.g. seminal
vesicles, coagulation glands)
Testes – stored in abdominal cavity before
maturation and ascends into scrotum after
sexual maturity
Prostate – consists of dorsal and abdominal
lobes

Reproduction
ANATOMY Sexing of pups - checking of the ano-genital
Skeletal System distance (female - shorter distance; male -
Two parts: axial and appendicular longer distance)
Dentition: I 1/1 C 0/0 P 0/0 M 3/3  

Digestive System
Esophagus is ~2 cm long; found at the back
of trachea; lined by a thick cornified
squamous epithelium
Stomach – proximal and distal portions are
keratinized and glandular, respectively;
capacity is 1.0-1.5 mL
Intestine – relatively shorter than herbivores
(e.g. rabbits, guinea pigs)  
Cecum - undeveloped Estrous cycle: proestrus, estrus, metestrus;
can be identified through vaginal cytology
Respiratory System 4-5 day estrous cycles
Right lung – divided into four lobes Ovulation – end of metestrus
(superior, middle, inferior and post-caval) Vaginal plug – indication that a female mice
Left lung – single lobe has been mated; typically lasts for 16 -24 hrs
Trachea – composed of 15 rings of cartilage but may reach 48 hrs
Trachea and bronchi – not robust; not Sexual maturity – 6-7 weeks of age; at 36
recommended for experiments on chronic days, sperm can be present in epididymis of
bronchitis or asthma drugs males; at 37 days, ovulation can occur in
females
Cardiovascular System Breeding – may start at 65-90 days
Heart – 4 chambers (left atrium, right atrium, Gestation – 19-21 days
left ventricle & right ventricle) Lactation – 20-22 days
Apex cordis – located near the end of Litter size – 6-15 pups (6-9 litters per year)
sternum and fourth intercostal rib; puncture
site for exsanguination MATING
1. Put the male and female mice (should be
Lymphoreticular System in proestrus stage) in same cage
Lymphatic system is highly developed 2. Allow to mate for 1 day
Lymph vessels, thymus, lymph nodes, 3. Check for the presence of vaginal plug
spleen, solitary peripheral nodes, Peyer’s
patches SIGNS OF PREGNANCY
No palatine tonsils 1. Increased food intake
Spleen contains megakaryocytes and 2. Increase in size of abdomen
primary hematopoietic cells 3. Prominent mammary gland

Reproductive System SIGNS OF PARTURITION

Female: paired ovaries and oviducts, uterus, 1. Nesting behavior
cervix, vagina, clitoris and paired clitoral 2. Enlarged and prominent mammary glands
glands 3. Mammary secretions (1-2 days before
Ovary is surrounded by a membrane and not parturition)
connected to the abdominal cavity (prevents  
ectopic pregnancy) DISEASES OF MICE
Typically 5 pairs of mammary gland (3 in 1. Pinworms – Syphacia and Aspicularis;
cervicothoracic region and 2 in inguino- inhabits the intestine
abdominal Region) 2. Mites – affected animals show pruritus,
  alopecia and scratch wounds (may be
 complicated by secondary bacterial Two divisions: forestomach or cardiac
infection) portion (non-glandular) and corpus or
3. Mouse hepatitis virus pyloric portion (glandular)
4. Ectromelia Unable to vomit
5. Pneumocystis Liver is composed of four lobes: median,
right lateral, left lateral & caudate
Regenerative ability of liver; Kupffer cells
have phagocytic ability
Gall bladder is absent
Bile ducts from each lobe form the common
bile duct which enters the duodenum

Respiratory System
LECTURE 4: RATS Left lung – single lobe
Right lung – cranial, middle, accessory and
RATS caudal lobes
 Rattus norvegicus No adrenergic supply to the bronchial
 Order: Rodentia musculature
 Family: Muridae Bronchoconstriction – controlled by vagal
 Second most common mammal used in tone
the laboratory Trachea – located in esophagus ventrally;
 More than 130 species (Norway rat or R. consists of 24 U-shaped rings of cartilage
norvegivus & black rat or R. rattus arere Trachea and bronchus are underdeveloped;
the most common) not suitable for studying chronic bronchitis
and asthma
EXAMPLE
Cardiovascular System
Blood supply to the atria of the heart – extra
coronary from branches of internal
mammary and subclavian arteries
Heart – located on a midline in the thorax;
apex is near the diaphragm with its lateral
aspects bounded mainly by the lungs

Urinary System
Right kidney is more cranial than the left
(cranial and caudal edge at the level of L1
and L3 vertebra, respectively)
A rat’s kidney has only one calyx – makes it
  suitable in efficiently studying cannulation of
GENERAL BIOLOGICAL CHARACTERISTICS the kidney
 Similar in appearance with mice but larger
in size and body weight Reproductive System
 Adult body length – 18-20 cm Male – developed accessory sex glands
 Tail – covered with short hair and circular (seminal vesicles, bulbourethral gland,
horny scales; used as main mode of coagulation gland and prostate gland)
cooling Inguinal canal remains open throughout the
 Absence of sweat glands on the skin; life of the rat
present on claws Female – bicornuate uterus
 Good adaptability for new environment Chest and abdomen have two rows of three
 Foraging and mating occurs during early pairs of nipples
morning and nighttime Presence of os penis (male) or os clitoridis
 Docile and easy to capture; pregnant and (female)
lactating rats are more aggressive
Endocrinology System
ANATOMY Pituitary gland – adheres to lower part of
Skeletal System the funnel; can be siphoned off with a
Components: skull, vertebrae, sternum, ribs, pipette to provide a pituitary extraction
forelimbs and hindlimbs model
Dentition: I 1/1 C 0/0 P 0/0 M 3/3 = 16

Digestive System
 Loud noises might cause cannibalism or
convulsion
Light – proper timing should be used to have
correct diurnal variations in the light cycle in
closed breeding cages
Omnivores; feed at any time
More active at night

GROWTH AND DEVELOPMENT

LECTURE 5: HAMSTER
Newborns – hairless, closed ears and short
limbs HAMSTER
Weaning – 20-21 days Order: Rodentia
Factors affecting growth rate: Suborder: Myomorpha
❖ Strain Family: Cricetidae
❖ Nutritional status  There are 24 species of hamster, and these
❖ Environmental conditions animals come in a wide range of sizes. Fifth
❖ Lactation capacity most commonly used lab animal in US
❖ Reproductive parity of female behind mice, rats, rabbits and guinea pigs.
 A small rodent that is widely disturbed in
REPRODUCTION many places of the Eurasian continent
Testes – descend into scrotum 30 -35 days  Two types/Common Laboratory Hamster:
after birth Syrian or golden hamster (Mesocricetus
Sperm production – 45 -60 days auratus) and the Chinese hamster or black
Mating begins at 60 days; optimal age is 90 lines hamsters (Cricetulus griseus)
days Other: Djungarian (or dwarf)
Vagina – opens 70-75 days (coincides with hamster - Phodopus
first ovulation) sungorus
Optimum breeding – 80 days
  Europen hamster - Cricetus
4-5 day estrus cycle (proestrus, estrus,
cricetus
metestrus, diestrus) – diagnosis thru vaginal
cytology  More than 80% of biomedical research
Ovulation – end of metestrus uses the golden hamster.
Females in group can inhibit estrus
Estrus – induced by male or its excreta
Gestation – 19-23 days
Litter size – 6-12 pups

NUTRITION  Nocturnal (foraging and mating occurs at

 Good animal model for nutrition night)
 Vitamin A, vitamin E, vitamin K, riboflavin  Sensitive to temperature changes
and thiamine deficiency – can cause ❖ Hibernation occurs when T = 8-9°C
infertility, skin disease or bleeding
❖ Pups can freeze to death at T below
 Effective in storing fat-soluble vitamin B12
13°C
 Able to produce vitamin C
❖ Room temperature should be 22-
 Able to supply most vitamin B – through
25°C
coprophagy
 Humidity should be 40-60%
 Commonly used for studying Ca and P
 Plant-based diet
metabolism
 Well-developed cheek pouches are
present on both sides of mouth
LABORATORY MANAGEMENT AND HUSBANDRY
 Ferocious, aggressive and attack each
Stress due to improper management and
other often
husbandry – promotes occurrences of
infectious diseases caused by opportunistic
ANATOMY
pathogens
Skeletal System
Ring necrosis – narrowing of skin of the tail
Dental formula: (incisors 1/1, canines 0/0,
or toes due to dry air or relative humidity
premolars 0/0, molars 3/3) = 16
less than 40%
Incisors of golden hamsters grow throughout
Respiratory infections – dirty litter,
their life
overcrowding, poor ventilation leading to
Spine – composed of 43 or 44 vertebrae,
excessive ammonia; most common is
including 7 cervical vertebrae, 13 thoracic
Mycoplasma
 vertebrae, 6 lumbar vertebrae, 4 sacral reproductive cycle of the golden hamster is
vertebrae, and 13–14 caudal vertebrae approximately 15 (14–17) days, which is the
Hamsters have a skeletal system with 124 shortest in rodents.
bones.

Digestive System
Stomach – forestomach and glandular LECTURE 5: GERBILS
stomach
Liver has 6 lobes – two left lobes, three right Gerbils
lobes, and a small middle lobe Order: Rodentia
Small intestine – 3–4 times as long as the Family: Muridae
body, 0.6 times the cecum, and 2.5 times the Sub-family: Gerbillinae
large intestine Scientific name: Meriones
unguiculatus
Respiratory System  There are over 30 different species
Lungs have five lobes - one left and four right  Excellent subjects for laboratory animal
lobes research
 small herbivores with ave. body weight of
Lymphatic System 77.9 (30 –113) g in adults
15 lymph organs and 35 –44 lymph nodes.  Mongolian Gerbil (Meriones unguiculatus)
 Males are bigger than females (body
length of 11.3 (9.7 –13.2) cm and ear
length of 1.5 (1.2 –1.7) mm )
Urogenital System  The front fur of the ear is gray, and the top
 Renal papillary is very long and extends inside fur is short and small (the rest of the
into the ureter ear is hairless)
 Testes (golden hamster) – large and  The hair is gray and brown on the back,
mulberry-like in shape; weighs 1.6–2 g lighter on the sides and cheeks, and gray
 Testes lie in the abdominal cavity on the on the abdomen
left side of the navel and under the  The tail is soft, long, and thick, and the
stomach. length is almost equal to the trunk,
 Uterus – “Y” shaped with a pair of round approximately 10.2 (9.7 –10.5) cm.
ovaries on each side.  Periods of activity alternate with resting
 Approximately 20 ova are produced from periods both during the day as well as the
one ovulation. night, but the highest activity takes place
 Female hamsters have 6–7 pairs of nipples. in the afternoon and night
 Male hamsters have a hip gland that  Their temperament is docile and fighting
produces secretions when in a state of does not usually occur, but mixed groups
sexual arousal of adult gerbils often lead to fierce fighting
accompanied by injury and death.
GROWTH AND DEVELOPMENT  
Sexual maturity (golden hamsters) – occurs COMON LABORATORY
between 30 and 32 days of age  Mongolian Gerbil (Meriones unguiculatus)
Sexual cycle lasts for 4–5 days  Fat-tailed Gerbil (Pachyuromys duprasi)
Growth and development of hamsters – 1–5  Pale (or Pallid) Gerbil (Gerbillus
years perpallidus)
Average life span – 2.5–3 years  Shaw's Jird (Meriones shawi)
Females are stronger than males and sexual
maturation is early ANATOMY
Skeletal System
REPRODUCTION Dental formula: 2 (incisors 1/1, canines 0/0,
Estrus cycle – proestrus, estrus, post-estrus, premolars 0/0, molars 3/3) = 16
and resting stage Spine – consists of 7 cervical, 12-14 thoracic,
Ovulation – occurs on the evening of estrus 5-6 lumbar, 4 sacral and 27-30 caudal
day 1 and can last into the night. vertebrae
Perennial estrus – animals with
characteristics of estrus after giving birth. Cardiovascular System
5–7 litters annually with 4–12 pups each.  Posterior communicating artery of Willis’
Newborn pups are hairless with closed eyes circle is deficient
and ears. The ears open after 5 days, while  There is no posterior communicating
the eyes open after 15 days, and the artery to connect the carotid artery system
weaning time is 21 days after birth. The
 and vertebral artery system, which cannot  Gerbils are generally housed in cages with
constitute a complete Willis’ circle a layer of sawdust. The cage should be
 Highly susceptible to cerebral infract shut tightly to prevent escape. Regular
following unilateral ligation of one cleaning of the cage needs to be carried
common carotid artery (can be an ideal out.
model to study the pathophysiology and  Separate feeding, replacing of bedding,
treatment of cerebrovascular disease such hybridization, and handling of gerbils
as ischemic stroke in humans) should be gentle. The use of solid-
bottomed cages is preferable for the
Reproductive System housing of gerbils. Gerbils often stand
 Large, ventral, sebaceous gland that is erect on their hind limbs, thus the height
androgen-dependent – secretes a special of the cage must be at least 15 cm. It is
smell important that cages have a solid bottom
 The male gerbil abdominal marking gland and the floor-to-lid height be tall enough
is larger and appears earlier than the to allow for this behavior.
female, -forming a hairless zone around  Feed protein content should be adequate,
the gland in adults. usually at least 16%, and can be
 The female abdominal marking gland is supplemented with fresh vegetables. The
small (difficult to find if not sheared), and breeding environment should be quiet and
its activity is enhanced during pregnancy ventilated because sudden noises may
and early lactation elicit epileptic seizures. Long-time
 In adult males, scrotal protrusions are cohabitation of males and females is
obvious, and have dark pigmentation preferred for breeding.
around the anus and scrotum LECTURE 5: GUINEA PIGS

Physiology Guinea Pigs

 Monogamous Order - Rodentia
 If the lighting regime is kept constant with Suborder - Hystricomporpha
12 h light, they can breed all year round Family - Cavidae
 Reproduction rates fall slightly in winter Genus - Cavia
 Sexual maturity: females - 9–12 weeks; Species - Cavia procellus
males - 10–12 weeks  Cavia porcellus from the family Caviidae
 Mean litter size: 3-7 pups  It resembles other cavies in having a
 Young gerbils suckle for approximately 21 robust body with short limbs, large head
days and begin to eat solid food at 16 days and eyes, and short ears
 Lifespan: 2-4 years (max of 14 births in  The feet have hairless soles and short
females under lab condition) sharp claws
 Newborn pups have no fur and weigh  There are four toes on the forefeet and
between 1.5 and 2.0 g with closed ears three on the hind feet
and eyes.  Weighs 500 g to 1500 g; body length is 20 -
 Ears prick up at day 3–4 and fur begins to 40 cm (8 -16 inches)
grow at day 6.  Tail is not visible externally
 At day 8–9, the incisors appear, and eyes  There is a crest of longer hairs at the neck,
open at day 16–18. but length and texture of the fur vary from
 Suitable mating age: 3–6 months smooth (short or long) to coarse and short
or long and silky
Reproductive Physiology  Coloration is extremely variable: the coat
 Sexual cycle: 4-6 days (vagina opens at 40- may be white, cream, tan, reddish or
60 days) chocolate brown, black, or a combined
 Sexual maturity occurs before 30 days pattern
 Gestation period: lactating females – 27  
days & non-lactating females -24-26 days COMMON LABORATORY GUINEA PIG
 Lactation – 14-29 days Hartley Guinea Pig
 Pups are weaned at 21 days after birth Coat Color: White (acromelanic albino)
(ideal weaning weight is 12 g) Ideal For: General multipurpose
 Behavior patterns of the two sexes are a model, surgical model
better indicator and a copulation plug is
generally not noticeable as it is small and The Dunkin Hartley is the most
retained deep inside the vagina common guinea pig strain used in
biomedical research, particularly for
Management and Husbandry studies of asthma, allergy, infectious
 disease, reproduction, and Lymphoreticular System
osteoarthritis.  Well developed lymphoreticular system;
extremely sensitive to pathogenic
Skinny Pig microorganisms
Coat: Hairless  There are many mononuclear leucocytes
Lifespan: 5–7 years carrying a granular structure (Kurloff body)
Fur type: Mostly hairless in the blood of guinea pigs.
 The Foa–Kurloff or Kurloff cell is an
The modern strain of skinny pig estradiol-dependent mononuclear
originated from a cross between leukocyte with natural killer activity that is
haired guinea pigs and a hairless lab found unique in the guinea pigs.
strain. In 1982 they were sent to
Charles River Laboratories to be bred Nervous System
for laboratory use and are commonly  Brain matures in embryos between 42 and
used in dermatology studies today. 45 days
   The cerebral hemispheres have no obvious
curve, with only a deep groove belonging
ANATOMY to smooth brain tissue, and is more
Skeletal System developed compared with other similar
 Dental formula: 2 (incisors 1/1, canines animals.
0/0, premolars 1/1, molars 3/3) = 20
 They have 32 to 36 vertebrae; the Reproductive System
vertebral formula is C7, T13(14), L6, S2(3),  Males have one pair of nipples in the
Cd4(6). inguinal region, same as females, but the
 Transversely inclined molar nipples of the female are slender and
 The incisors are arcuated and extend to located above the glands
the jaw  Accessory sex glands in the male guinea
 There is diastema or gap between the pig include large transparent, smooth
incisors and premolars seminal vesicles, prostate, coagulating,
 All teeth are open rooted and grow bulbourethral, and rudimentary preputial
continuously glands
 Testes remain in inguinal pouches with
Gastrointestinal System inguinal canals open for life
 The oral cavity is small and narrow, making  Adult males have large testes in obvious
endotracheal intubation difficult and the scrotal pouches.
stomach wall is very thin, with a capacity  Females have a U-shaped depression in
of 20–30 ml the perineal tissue.
 Monogastric stomach – lined entirely with  The anus is located at the base of the U,
glandular endothelium and the branches are covered by a
 Intestine is 10x longer the body length membrane.
 Cecal fermenter; cecum can hold up to  The uterus is bicornuate in females, which
65% of the total gastrointestinal contents. terminates into a single cervix. The vagina
is closed during estrus
Respiratory System
 The trachea and bronchi are
underdeveloped
 Tracheal glands are only located in the
throat
 The guinea pig can be used as a model for
lung-function impairment and bronchial
reactions including airway hyper-
responsiveness and reactions that
resemble asthma in humans
 The guinea pig has the most prominent
smooth muscle in the distal bronchi and
muscle is spirally arranged
 The lung is divided into seven lobes (the
larger right lung has four lobes (upper,
middle, lower, and side lobe), while the
left lung has three lobes (upper, middle,
and lower lobe))