1.

1 INTRODUCTION

The Albino rat (officially known as the Pink-Eyed White or PEW) is most
likely the very first mutation to be discovered and purposely bred. Albinism is a
very common mutation even in wild populations, showing up in many animals
including dogs, horses, ferrets, rabbits, cavies, and many others including rats
and mice.

Albino rats have impaired vision as compared to normally pigmented rats.

Albino rats have fewer light sensitive cells in their eyes, an abnormal eye-to-
brain connection, and a translucent iris which leads to dazzling and retinal
degeneration. Albino rats also take much longer than pigmented rats to adapt to
low-light conditions. As a consequence of all these differences, albino rats are
severely visually impaired or blind.

Albino rat, Rattus norvegicus, belongs to the order Rodentia and family
Muridae. Albino rats were first used for experimental purposes in the mid-
1800s. Strains were developed to study neuroanatomy, nutrition, endocrinology,
genetics and behavior.

The rat has short hair, a long naked tail, rounded erect ears, protruding eyes, a
pointed snout with long whisker (vibrissae) and five toes on each foot. Albino
rats have poor eyesight and depend on facial vibrissae and olfaction for sensory
input. Albino rats have no gall bladder. Rattus norvegicus (brown rat) has a
short tail compared to Rattus rattus (black rat), which has a much longer tail.

Albino rats have a pair of incisors and three pairs upper and lower of molars.
Molars are permanently rooted while the incisors have an open root and grow
continuously. Due to this continuous growth of the incisors rats can have

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problems with incisor overgrowth when the upper and lower incisors do not
meet properly (malocclusion). Malocclusion can be hereditary or follow trauma,
disease or inappropriate diet and/or soft food. There is no permanent cure for
overgrown teeth. The only treatment is to trim the teeth every 2-3 weeks, if
malocclusion persists.

Rats have a large horseshoe-shaped Harderian gland deep within the orbit.
Secretions from the gland contain varying amount a reddish-brown porphyrin
pigment depending on the physiologic state, age, strain and sex of the rat. The
amount of secretions increases during stress and appears as 'red crusts' around
the eyes and nostrils.

1.2 BASIC REQUIREMENTS FOR HOUSING AND HUSBANDRY

Basic requirements for good rodent housing and husbandry are given in the
table below.

Housing in stable, compatible groups - it is important to take into account sex,

age, reproductive condition, familiarity, prior group housing experience when
grouping the animals.

Enclosures designed to cause minimum disturbance to the animals.

Enough space for exercise, normal social behaviour (e.g. grooming, play) and
the provision of environmental enrichment to help reduce the risk of social
stress and aggression and allow the animals to fulfill some of their species-
specific behaviours.

Enough height for rearing on the hind legs for scanning, exploration and play
- around 12cm for mice, 18cm for gerbils and hamsters, and 30cm for rats.

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Solid floors with an adequate depth of an appropriate substrate (e.g. 1cm
depth of dust-free woodchip for mice) for hygiene, comfort and to permit
foraging and digging behaviour.

Material to gnaw (e.g. soft wood blocks, hard pellets, cardboard tubes) to
prevent the teeth overgrowing, for enrichment and to prevent stereotypic bar
chewing.

Refuges (e.g. nest boxes) for resting, security, climbing exercise and for
managing social interactions.

Vertical barriers or tubes (e.g. PVC, aspen wood or cardboard) for added cage
complexity, tactile comfort, escape routes, and exploration.

Nesting material (e.g. soft paper or soft wood) for comfort, to help regulate
temperature and light levels, and to hide and retreat from cage mates or
threatening stimuli. Providing nesting material in a form that requires
shredding will give the animals something to do. Nesting material is essential
for parturient females. Nest boxes should be provided if insufficient nesting
material is provided for the animals to build a complete, covered nest.

Appropriate lighting levels and regimes. Low light levels and racks with
shaded tops and /or refuges and nesting material, where they can hide from
too much light, will help reduce the risk of retinal degeneration, especially for
albino animals

A varied diet and the ability to forage - scattering food (e.g. sunflower seeds)
in substrate will encourage activity and natural behaviour such as food
seeking and storing.

Minimisation of extraneous noise and ultrasound.

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 Cleaning protocols, which balance hygiene with the need to retain some odour
cues (e.g. scent-marked nesting material) to avoid stress and aggression.

Gentle and frequent handling from early in life.

Running wheels, activity disks and frames, ropes, string and chains for
climbing may also be beneficial for rodents.

Whenever enrichments are provided, these should be in sufficient number and

at a sufficient distance so that aggressive competition is not triggered.

Individually ventilated cages

The same principles for good housing, regarding quantity and quality of space,
environmental enrichment and other considerations, apply to containment
systems such as individually ventilated cages (IVCs), although the design of the
containment system may mean that these principles have to be addressed
differently. The draught induced by high intra cage-ventilation rates in some
IVCs can induce chronic stress and heat loss. The location of the air supply to
the cage (e.g. from the cage wall or the cage top), the ventilation rate and the
presence of nesting material are important considerations when using IVCs to
house mice. Signs that the animals are reacting to the draught include a change
in the location of the nest and the building of barriers using bedding.

Reproduction

Albino rats have two distinct cervices and uterine bodies. There are separate
urethral and vaginal openings. There is a vaginal closure membrane, which is
lost at puberty. The inguinal canal remains patent throughout life. Albino rats
have an os penis or os clitoridis associated with external genitalia. Mammary

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tissue is widely distributed and may extend to the lateral and dorsal areas of the
abdomen.

Albino rats have a four to five-day estrous cycle, divided into characteristic
phases: proestrus, estrus, metestrus and diestrus. The stage of the estrous cycle
can be determined by vaginal cytology. Ovulation occurs at the end of
metestrus. Receptive females exhibit lordosis when a downward pressure is
applied to the pelvis. Such females will adopt a rigid posture with the
hindquarters raised. Mating leads to formation of a vaginal plug. Plugs persist
for 16-24 hours and may last as long as 48 hours.

Albino rat vaginal smear

Samples for making a vaginal smear can be collected by inserted a cotton tipped
swab moistened with phosphate buffered saline into the vaginal cavity of a rat.
The swab should be applied gently against the vaginal wall and rolled slightly
before withdrawing. The moist swab is then rolled onto a clean glass
microscope slide. The specimen is spray fixed using 95% ethanol. Air drying or
dipping the slides in 100% alcohol does not produce but the very good cytologic
preparation of rat vaginal smears. Samples for making a vaginal smear can also
be collected by introducing 1.0 ml of saline into the vaginal cavity using a blunt
tipped disposable pipette, and recovering .25 ml of vaginal fluid a short time
later. This material is then applied onto a microscope slide and processed as
above. Fixed specimens are then stained using Papanicolaou stain for best
results and examined using a light microscope under low power.

Pregnancy

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Pregnancy lasts 21-23 days. Females will build a nest prior to parturition if
opportunity is provided. Birth usually occurs at night with 10-12 pups being
born. Stretching and hindleg extension are usually signs of impeding birth.
Babies are born either head or tail first (breech). The female usually eats the
placenta. Delivery lasts 1-4 hours, if labor persists call a veterinarian (5-3713).
There is a fertile postpartum estrus. Maternal antibody is transferred to the fetus
in utero and to the newborn via colostrum.

Husbandry

Rats are generally fed a diet containing low fiber (5%), protein (20%) and fat
(5-10%). Feed may be pelleted or powdered. The pelleted feed is supplied as
regular, breeder, certified, irradiated or autoclavable. Albino rats are usually
supplied feed free choice and they eat 10-30 g a day (5 g/100 g body
weight/day). Water is supplied free choice and they usually drink 20-50 ml a
day (10 ml/100 g body weight/day). Water may be supplied using a bottle or
automatic waterers, and may be further treated by reverse osmosis, ozone,
ultraviolet radiation, hyperchlorination or acidification.

Albino rat rooms are usually maintained at 30-70% relative humidity and a
temperature of 18-26ºC with at least 10 room air changes per hour. Albino rats
are usually housed in standard shoebox cages with or without filter tops. Filter
tops prevent cross contamination of rats limiting the spread of disease and keep
facilities clean. Cages with filter tops may have a slightly higher temperature,
relative humidity, carbon dioxide and ammonia than the room air. Microisolator
tops provide even a higher level of protection than bonnet type filter tops, since
they seal better. Static cages as described above are usually changed one to two
times a week depending on cage density and housing style. In ventilated cages
air is forced into the cage at up to 60 air changes per hour. This keeps the cage

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dry and reduces build up of ammonia and carbon dioxide. In such situations
cages are changed once every 1-2 weeks. Ventilated cages may be kept positive
or negative to room air depending on the study being performed.

Albino rats are usually provided with bedding in the shoebox cages. Bedding
can be paper, wood shaving, wood chips or corncob. In very rare instances
Albino rats are housed on wire floors. Housing on wire floors must be justified
and approved by the animal care and use committee.

Identification

Albino rats should always be clearly identified on cage cards indicating protocol
number, strain (using standard nomenclature), sex, age, supplier, investigator
and contact person. A combination of marks made by an ear punch can be used
to identify rodents as indicated in the figure.

Procedures performed on the animal should be clearly indicated. Individual rats

can be identified using ear punches, ear tags, tattoos, fur dyes, indelible mark on
tail or microchips.

Sexing

Sex is determined using the anogenital distance. Males have a greater (1.5-2
times) anogenital distance than females as well as a larger genital papilla. In
neonatal males the testis may be visible through the abdominal wall.
Conspicuous bilateral rows of nipples are visible in females at about 9 days of
age. Absence of testicles is not a useful criterion for sexing since the testis is
retractable throughout life into the open inguinal canal.

Handling

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Albino rats should be acclimatized to handling (gentling) to reduce stress.
Always talk quietly, move hands slowly and handle them frequently. Albino
rats should be handled at the base of the tail using your fingers. Pick up rats by
placing the hand firmly over the back and the rib cage and restraining the head
with thumb and forefinger immediately behind the mandibles. Holding the rat
upside down keeps it distracted and reduces the chances of biting. A variety of
restraint devices are available to assist in handling rats.

Blood collection

An adult Albino rat has a circulating blood volume of about 15-35 ml (5-7% of
the body weight), however in older and obese animals this may be lower. Up to
10% of the circulating blood volume may be taken on a single occasion from a
normal healthy animal on an adequate plane of nutrition with minimal adverse
effect. Always make sure the animal has recovered safely from the procedure
and give warm isotonic fluids. This volume may be repeated after 3-4 weeks.
For repeat bleeds at shorter intervals, a maximum of 1% of an animal's
circulating blood volume can be removed every 24 hours.

Blood can be collected from several sites in the rat including tail vein,
saphenous vein, retro-orbital sinus, brachial vessels, vena cava or cardiac
puncture. Always ensure complete hemostasis before returning the rat to its
home cage.

Tail

It may be necessary to warm the tail by exposing it briefly to a heat lamp or

placing it in a bowl of warm water. The Albino rat should be restrained in a
device for the collection. Blood can be collected from the tail vein (and artery)

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by making a snip in terminal =5 mm of the tail with a scalpel or sharp scissors.
Stroke the tail gently with thumb and finger to enhance flow of blood into the
collection vial. Because of the thermoregulatory function of the tail no more
than the distal 3 mm should be taken at a time. At the end of the collection
apply pressure to the cut end with a gauze bandage and ensure that blood has
completely stopped flowing before returning the rat to the cage. A small nick
can also be made at side of the tail 0.5 -2cm from the tail base to collect blood.
A fine gauge needle introduced through the skin at a shallow angle can be used
to withdraw blood from the tail vein. Apply a tourniquet around the base of the
tail to aid in the collection. A butterfly catheter with only about 5 mm of tubing
attached to it (rest cut off) may be used instead of a needle and syringe.

DISEASES OF ALBINO RATS

Diseases of Albino rats are usually handled as a herd (colony) health problem
rather than on an individual animal basis. In most cases the goal is to prevent
introduction of a disease into a colony than to treat individual animals. Disease
prevent is practiced by institution of a health monitoring (sentinel) program
based on serological and microscopic diagnosis of problems in a representative
sample of animals. Due to the widespread movement of animals all over the
world with advent of genetic manipulation of animals, the possibility of
introducing disease agents in a colony has markedly increased. The expanded
use of genetically modified and immunocompromised animals greatly
exacerbates the problem. Furthermore the practice of transplanting tumor
material into animals provides a portal where these agents can be introduced
into animal, especially if the tumors are not screened for adventitious infectious
agents. Some important rat diseases are discussed below to draw attention to the

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need to adhere to practices recommended by the veterinary staff to avoid these
diseases.

Pinworms

Pinworms (Syphacia and Aspicularis) inhabit the intestine (cecum, rectum,

colon) and have a direct lifecycle. The eggs are particularly resistant and
survive for a long time in the environment. The disease is usually subclinical
being marked in weanlings and immunocompromised animals. Symptoms
include poor body condition, rough hair coat, reduced growth rate and rectal
prolapse. Infection with pinworms has a negative impact on gastrointestinal,
growth, behavioral and immunology studies.

Mites

Mites affect the skin of rats and up to 100% of the animals may be affected.
Affected animals are scruffy, pruritic (itchy), loose hair and have scratch
wounds, which can become infected with bacteria. There are changes in the
immune responses of affected animals.

Sialodacryoadenitis virus

Sialodacryoadenitis virus (rat corona virus) affects the salivary and lacrimal
glands and the respiratory tract of rats. The virus spreads extremely rapidly in a
susceptible population resulting in conjunctivis, nasal and ocular discharge,
porphyrin (red) staining around the eyes and nostrils, corneal ulceration,
swelling of the neck and protrusion of the eyes (exopthalmus) resulting in
drying of the cornea. There are changes in the respiratory passages (squamous
metaplasia) during recovery from the disease. The disease may persist for at
least 6 months in immunocompromised animals. The disease interferes with

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growth and reproduction in rats, and adversely impacts on studies of respiratory,
ocular, olfactory and immune systems. It adversely affects studies involving
infectious agent especially in the respiratory system as well as tumor studies.

Pneumocystis

This is a fungal disease affecting a wide range of laboratory animals and

humans. The organs are primarily localized in the lungs but may also involve
other organs including the eyes, skin etc. It causes a slowly progressive chronic
pneumonia with weight loss and eventually death in a large number of
immunocompromised animals. The disease has a severe negative impact in
research involving immunocompromised animals, pulmonary function and
immunology.

REFERENCES

Baumans V (2005) Environmental enrichment for laboratory rodents and

rabbits: requirements of rodents, rabbits and research. In: Enrichment Strategies
for Laboratory Animals. ILAR Journal 46: 162-170.

Burn CC (2008) What is it like to be a rat? Rat sensory perception and its
implications for experimental design and rat welfare. Applied Animal
Behaviour Science 112: 1-32.

Gaskill BN, Gordon CJ, Pajor EA et al. (2012) Heat or insulation: Behavioral
titration of mouse preference for warmth or access to a nest. PLoS ONE 7(3):
e32799.

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Harris A, D'Eath RB, Healy SD et al. (2009) Environmental enrichment
enhances spatial cognition in rats by reducing thigmotaxis (wall hugging)
during testing. Animal Behaviour 77: 1459-1464.

Reinhardt V, Reinhardt A, Eds. (2002) Comfortable Quarters for Laboratory

Animals, 9th Edition, Animal Welfare Institute.

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