Welcome to the course Writing an Animal Protocol for Research on Cats.

This is the cat module in a course series on the preparation of an animal use protocol. Each course in this series refers to a different animal species. Every course offers information that is both pertinent to all research animals and specific to the one animal species presented.

• The first 7 lessons are similar in all courses of this series. Differences in these lessons relate mainly to the regulatory coverage, housing requirements, and the zoonotic hazards of each animal species.

• The remaining lessons present information that is more specific to each animal species, such as biological features, anesthetic doses, and biomethodologies.

The goal of this course is to cover important information about using cats in biomedical research settings. If you are responsible for handling cats or if you must write an animal use protocol, this course will be useful by providing you with::

• Information on key regulatory issues.
• Guidance on searches for alternatives in the care and use of animals.
• Highlights of unique biological features of these animals.
• Overviews of acceptable basic methodologies.
• Requirements for supportive care procedures.

Hypertext links in this course provide you with supporting information, such as regulatory sources, drug doses, practical tips, etc.

This course will not provide you with detailed information on how to conduct the methods and procedures described. For this, you should use other courses offering in-depth information and hands-on instruction from your institution's animal facility staff.

Click here to view the credits for this course.

To ensure the humane treatment of laboratory animals, research involving cats is regulated by two federal agencies:

• The United States Department of Agriculture (USDA) / Animal Care; and
• The Public Health Service (PHS) / Office of Laboratory Animal Welfare.

If your institution receives any funding from the PHS or is voluntarily accreditated by the Association for Assessment and Accreditation of Laboratory Animal Care, International (AAALAC), then your research must also comply with the National Research Council publication, the Guide for the Care and Use of Laboratory Animals. This document will simply be referred to as the Guide in this course.

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

It is the responsibility of principal investigators to assure that their laboratory staff are informed of and participate in their institution's occupational health and safety program.

Elements of an occupational health and safety program, including institutional responsibilities, are described in the guideline, Occupational Health and Safety in the Care and Use of Research Animals (shown at right), published by the National Research Council.

Working with cats is associated with the following hazards:

Injuries
Personnel handling cats can be injured by bites or scratches.

• Cats have long, needle-like canine teeth and sharp carnassial teeth.
• Wounds caused by deep penetration of the canine teeth may become infected with dangerous pathogens such as Pseudomonas spp. and Pasteurella spp.
• Their toenails can inflict painful scratches as when a cat is struggling against restraint.

Injuries are more likely when personnel lack the knowledge of how to handle, transport, and restrain a cat. Likewise, poor technique in handling, etc., can cause injury to the cat. Training staff to work effectively and humanely with cats is essential to prevent injuries to people and cats.

Allergies
People can develop an allergy to cats after having contact with them for some time. Persons who develop allergy symptoms should seek medical counseling and may have to discontinue working with this species.

Zoonoses
In general, transmission of zoonotic disease from naturally infected laboratory animals is uncommon because of the ongoing efforts to improve the health status of animals by vendors and by facility staff. Health surveillance programs, routine sanitation, and personal protective equipment have important roles in preventing zoonoses. The photo shows an early Bartonella henselae lesion on the thumb of a handler.

Cats can be a reservoir of the following infectious agents which are transmissable to people:

Viruses

Rabies virus

 

Ricksettia

Bartonella henselae

 

Bacteria

Campylobacter jejuni

Leptospira spp.

Salmonella spp.

 

Fungi

Microsporum spp., Trichophyton spp.

 

Protozoa

Cryptosporidium spp.

Giardia spp.

Toxoplasma gondii

For more information, refer to Occupational Health and Safety in the Care and Use of Research Animals, published by the National Research Council.

Your protocol form should ask you for an assurance that you have considered alternatives to the use of animals if painful or distressing procedures are proposed. This is to satisfy mandates by the Animal Welfare Act and PHS Policy to avoid or minimize discomfort, pain, and distress consistent with sound scientific practices. Alternative procedures are those which may replace animals with nonanimal methods, reduce the number of animals used, or refine the methodology to minimize animal pain or distress.

For more information on what is meant by alternatives to the use of animals, please refer to the course Working with the IACUC, which is part of this series.

The assurance must take the form of a written narrative that describes which sources were used to determine that alternatives were not available. Typically, you may be asked to provide the results of a database search including information on:

1. The databases searched.
2. The date the search was performed.
3. The years of citations covered by database searches.
4. The key words and/or search strategy used when searching a database.

It is strongly recommended that this information be sought during development of a protocol.

Organizations that can assist you in performing an alternatives search are:

• ALTWEB, Center for Alternatives to Animal Testing, Johns Hopkins University
• Animal Welfare Information Center, National Agricultural Library
• Center for Animal Alternatives, University of California – Davis

The following is a case study of alternatives searches that may guide you in the development of a search strategy that is pertinent to your own research.

Click on each database below for a sample search on key terms for this example.
You can follow the links to the database for a search in real time.

Example Search:
Study of auditory response in the cerebral cortex of cats.

PubMed

Agricola

BIOSIS

More Example Searches:
Additional search routines are available elsewhere in this course to provide you with guidance on performing searches on alternatives in the care and use of animals related to the lesson topic.

All of the federal laws, regulations, policies and guidelines applicable to animal research have as their core intent the humane treatment of the animals involved in a study. Accordingly, your IACUC will have requirements for the proper care of your animals prior to, during and after a research procedure.

What is a procedure? A procedure is any activity carried out on the animal, such as venipuncture or surgery. This peri-procedural care requirement includes:

• Properly preparing the animal to humanely undergo the procedure;
• Supporting the animal's physiological function during the procedure; and
• Providing additional supportive care to aid the animal in recovering from the procedure.

The images below and on the next screen shows a cat in a restraint method for a nonpainful procedure, such as venipuncture. Below, a cat wrapped in a towel. On the next screen, a cat in a zippered cat-bag.

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

When performing any procedure, such as a blood collection, you should think through the steps that are necessary to protect the animal's welfare. For example, for blood collection, you should limit the volume to the minimum that may be taken safely. That amount may vary depending upon whether a single sample is taken or multiple, chronic collections are done and, with any venipuncture, you should be prepared to care for the animal in the event of trauma to the vein area or excess hemorrhage.

Refer to your institution's IACUC and attending veterinarian for specific guidelines.

Cats may be housed singly or in pairs or groups, and in cages or pens. Housing cats together in the same enclosure is often preferred because cats, providing they are properly socialized, engage routinely in social activities to sleep, groom, and play.

When housed together, compatibility among cage mates is an important consideration for minimizing inter-animal aggression. Females in estrus should not be housed with males except for breeding.

Federal regulations provide standards for cat housing.

• Cages or pens should allow for normal bodily function and protection from injury.
• The animal should be comfortable and free to make normal postural adjustments and movements.
• Housing should be secure, durable and in good condition.
• Cages and pens should be constructed to facilitate cleaning and sanitation.
• Cages should have smooth surfaces that are impervious to moisture and have no crevices which accumulate soil.

Cats are intelligent animals that easily become bored when caged or housed indoors. It is common, therefore, to provide cats with enrichment devices, e.g., nontoxic toys that can be tossed or dangled to simulate the motion of prey.

Example Alternatives Search:

For additional information on types of cat housing, please refer to the examples of "alternatives" searches on cat housing.

Cats used in research must be obtained from sources authorized by the USDA. Laboratory cats are either bred specifically for research (purpose bred) or are obtained from various sources (random source).

 

Purpose bred cats, bred by USDA-licensed dealers specifically for research. Such animals have a clearly defined health, pedigree and vaccination status. Commercially available cats are of the breed Domestic Shorthair (DSH). The cost of purpose-bred cats may be higher because of the expense of raising healthy cats to adulthood. Their health status is generally better than that of unconditioned random source cats.

 

Random source cats, which are obtained either from another class of USDA-licensed dealers or from animal pounds and shelters. These animals usually have no vaccination history and have poorer health. Prior to being used in an experimental protocol, they should be physiologically stabilized during an acclimation period. This involves treating diseases and parasites, immunizing the cats and improving their plane of nutrition. In many areas, local ordinances limit the use of pound and shelter cats for research.

Many research facilities have a policy to use only purpose bred animals in order to provide a high quality of animals for research and to avoid the health and ownership issues associated with random source animals.

Upon arrival at your facility, your cats should have an acclimation period before they are used in research studies. This period of time allows animals to adapt to a new environment. Effects of transportation stress include alterations in various blood parameters, immune cell function and animal behavior. The period of time necessary for biological stabilization will depend on the parameters to be studied.

Refer to your institution's attending veterinarian for recommendations that are appropriate for your project. Typically, acclimation periods range from 4 days to 1 week.

Example Alternatives Search:
For citations on the impact of transportation stress on cats, please refer to the example of "alternatives" searches provided.

Routine quarantine procedures may prolong the holding of your animals in special facilities. Quarantine aims to prevent transmission of diseases between new animals and established colonies.

Acclimation and quarantine periods run concurrently, although they serve different purposes. Most institutions do not allow experiments on animals while quarantined.

Example Alternatives Search:
For citations on quarantining cats, please refer to the example of "alternatives" searches provided.

Cats arriving in poor health, such as is the case with many random source cats, undergo a period of conditioning for treatment of parasites and infections. Treatment regimens should attain an animal health status that is compatible with the experiments in which the animals will be used.

The conditioning period, which runs concurrently with the acclimation and quarantine periods, will vary in duration with the types of treatments needed. Conditioning periods may extend from 14 to 30 days.

If your proposed study involves a painful procedure, the protocol form may ask for a method of assessing if the cats are experiencing pain or distress.

The signs of acute pain and distress can vary according to the nature and location of the pain, the animal’s pain threshold, emotional state, and environment. You must know how cats appear and behave normally in order to recognize abnormalities that may signal pain and distress.

In the cat, signs of pain or distress may include changes in gait, activity, posture, attitude, and even facial expression.

• Cats will cease to groom themselves and their fur will become scruffy or greasy.
• Cats will continuously lick, and even mutilate, a body part if they feel pain or abnormal sensations.
• Cats may vocalize or become aggressive if a sore area is touched or moved.
• Cats often retreat and hide when in pain or distress; this behavior emphasizes the importance of checking cats that are withdrawn from the colony or from human contact.
• They may also reduce their intake of food and water.
• Cats may lose body weight and deteriorate in their overall condition.
• Purring is not always a sign of well-being in cats because they may purr when severely ill.

Behavioral signs of acute pain and distress in the cat are:

• Hyperactivity, restlessness, nervousness, bouts of frenzied activity, anxious facial expression, self-mutilation or continuous licking of a body part.
• Hypoactivity, listlessness, reluctance to move or get up, hiding and retreating.
• Altered gait, favoring body part.
• Self-mutilation or licking.
• Aggressiveness when approached or touched.
• Vocalization – howling or growling either when unprovoked or touched; also may be silent.
• Reduced intake of water and food.
• Loss of body weight.
• Cessation of grooming and changes in fur coat – dull, fluffed or greasy.

A chronic state of pain or distress may be more subtle and difficult to detect. A good knowledge of the animal’s normal appearance and behavior is especially important to recognize chronic pain or distress.

Example Alternatives Search:
For citations on detecting pain and distress, please refer to the example of "alternatives" searches provided.

A knowledge of the characteristics of cats is helpful to effectively manage these animals and to plan experimental procedures for their use. Researchers should be aware of the following practical features. Click on the following items for a brief description and some practical tips.

 

Behavior

Social behavior among cats

Socialization to humans

Communication cues - dominance and submissiveness

Assessing a cat’s response to you

Safely approaching a fearful cat

 

Gastrointestinal

Nutritional requirements

Digestive System

Teeth

Pre-surgical fasting

 

Pharmacological

Ketamine adverse effects

Drug metabolism and toxicity

Sensitivity to opioids

Below are volume recommendations for acute intravenous fluid administration and blood collection in cats.

^aRemoving greater quantities of blood (exceeding 10% of total blood volume) can produce hypovolemic shock. Repeated collections of smaller amounts of blood will have the same effect. In such procedures, animals should receive warmed, physiological fluids to replace the volume of blood collected. In addition, monitor the animal’s hematocrit for anemia.

From:

1. Hawk and Leary, Formulary for Laboratory Animals, 2nd Edn., Iowa State University Press, 1999.
2. Wolfensohn and Lloyd, Handbook of Laboratory Animal Management and Welfare, 2nd Edn., Blackwell Science. 1998.

Below are peripheral vessels that are commonly accessed for blood collection or fluid administration. Recommended needle sizes are 21 to 25 gauge. Of this gauge range, the larger needles may be necessary for injecting large volumes or viscous materials. The photo shows the location of the medial saphenous vein.

1. Jugular vein
2. Cephalic vein
3. Medial saphenous vein

Below are the nonvascular routes of injection that are commonly used in cats. Included are volume recommendations for the acute administration of fluids. Recommended needle sizes are 21 to 25 gauge; larger needles may be necessary for injecting viscous materials.

Subcutaneous (SQ or SC)

55-100 ml (scruff, back)

 

Oral (PO)

3 ml/kg

 

Intramuscular (IM)

1 ml per site

From:

1. Hawk and Leary, Formulary for Laboratory Animals, 2nd Edn., Iowa State University Press, 1999.
2. Wolfensohn and Lloyd, Handbook of Laboratory Animal Management and Welfare, 2nd Edn., Blackwell Science. 1998.

For citations of blood collection procedures in cats, please refer to the examples of "alternatives" searches provided.

This section includes cat dose rates for the common drugs and drug regimens. If you need to use other drug agents, check with your institution's veterinary staff for assistance in determining a dose rate appropriate for use in cats.

Below, a kitten becomes anesthetized by mask delivery of a gaseous agent.

Click on the drug types for doses of common agents and drug regimens:

Postoperative Analgesics:
Available in two classes of drugs – the opioids and the nonsteroidal anti-inflammatory drugs (NSAIDs).

Sedatives:
Sedatives may obtund consciousness but in normal doses do not do so sufficiently to ablate the perception of pain or other sensations. When combined with general anesthetics, they may be used to induce a "balanced" anesthesia where muscle relaxation, unconsciousness, and analgesia are enhanced.

Sedatives + Analgesia:
Some sedatives also have analgesic effects. When combined with general anesthetics, a balanced anesthesia is attained, and these sedatives enhance analgesia through specific effects.

Anesthetics: Gaseous and Injectable
An anesthetic regimen should be chosen to match the duration of drug effects with the length of the procedure. In particular, short acting agents (and regimens) should be not be used for long procedures because repeat drug administrations, necessary to prolong anesthesia, will produce uneven blood concentrations and therefore periodically inadequate anesthesia. For long procedures, gaseous anesthesia is the often the most practical method to sustain uniformly adequate levels of anesthesia.

For additional information on analgesia and anesthesia in cats, including the physiological impact of these agents on cats, please refer to the examples of "alternatives" searches provided.

Aseptic technique should be used when performing major survival surgery on cats. This is required by the Animal Welfare Act (federal law). The standards described here are consistent with the Guide for the Care and Use of Laboratory Animals.

 

Survival surgery means that the animal regains consciousness after anesthesia. In nonsurvival surgery, the animal is euthanized before recovery from anesthesia.

 

Major surgery means penetrating and exposing a body cavity such as the chest or abdomen; or producing substantial physical or physiological impairment.

If you will be performing major survival surgery on cats, federal requirements are that:

• The surgery be carried out in a dedicated surgical facility.
• The surgeon must wear cap, mask, and sterile gown and gloves.
• The incision site is appropriately clipped, scrubbed, disinfected, and draped.
• Instruments and surgical materials are sterile.
• The animals receive proper supportive care and monitoring for anesthesia and vital signs through the procedure and afterward.

If you will be performing minor survival surgery, requirements are less stringent than outlined above. According to the Guide, requirements still include: "aseptic technique and instruments and appropriate anesthesia. Although laparoscopic procedures are often performed on an 'outpatient' basis, appropriate aseptic technique is necessary if a body cavity is penetrated."

If you will be performing nonsurvival surgery, it may not be necessary to follow all the techniques outlined above for major survival surgery. According to the Guide, "at a minimum, the surgical site should be clipped, the surgeon should wear gloves, and the instruments and surrounding area should be clean."

Supportive care aims to:

• Maintain the animal's physiological status as nearly normal as possible.
• Minimize animal pain and distress.

Supportive care includes the monitoring of both physiological parameters and analgesia during anesthetic and surgical procedures. Monitoring of vital signs and pain should be conducted throughout the procedure and the recovery period.

Keep in mind that:

• General anesthesia causes disturbances of thermoregulation and other physiological functions.
• Warming devices (e.g., heated tables and pads) are recommended for routine use to maintain the animal's body temperature. Care should be taken to avoid use of devices that may cause thermal injury.
• During surgery, the animal may experience pain if anesthesia is inadequate at any time during the procedure.
• Postoperatively, the animal may experience pain, discomfort, and distress unless treated with analgesics and appropriate supportive care measures.

Due to the interaction of metabolic factors and drug effects that can cause animal mortality, cats should receive good supportive care and monitoring during anesthesia, whether or not the procedure involves surgery.

During anesthesia and surgery, the following procedures are recommended.

Supportive Care

• Provide a source of warmth to a cat from the onset of anesthesia to the end of anesthetic recovery.
• Infuse sterile physiological fluid (warmed to body temperature) to compensate for blood loss during a procedure and depressed fluid intake post-procedure.

 

Monitoring during Anesthesia:

• Analgesia - toe pinch.
• Respiration - gross changes in rate, character of breathing.
• Color of mucous membrane and skin – poor oxygenation (blue), poor blood perfusion (pale)

After anesthesia and surgery, the following procedures are recommended.

Monitoring post Anesthesia:

• Cats must be monitored until fully recovered from anesthesia as indicated by the ability to ambulate and maintain core body temperature.

 

Monitoring post Procedure:

• Assess appearance, activity, and behavior as indications of pain and discomfort (see screen Detecting Pain and Distress).
• Assess food and water intake.
• Assess wound repair. Routine use of antibiotics is not indicated after uncomplicated aseptic surgery.

At right, a cat receives nutritional support by infusion with a nasogastric tube.

The term euthanasia is derived from Greek and means "good death". Animals should be euthanatized when killed for any purpose, including research. To euthanatize a cat, you must be trained in the concepts of euthanasia, the method to be used, and the proper handling of cats.

Methods are classified as acceptable or conditionally acceptable, as set by the American Veterinary Medical Association. The inclusion of conditionally acceptable methods in your protocol may require scientific justification and IACUC approval.

Below, an injectable agent which is accetable for euthanasia of cats.

Click on each method of euthanasia below for recommendations on its use in euthanatizing cats.

 

Acceptable Methods:

• Barbiturates
• Inhalant anesthetics
• Carbon dioxide
• Carbon monoxide
• Potassium chloride in conjunction with general anesthesia

 

Conditionally Acceptable Methods:*

• Nitrogen
• Argon

* The inclusion of conditionally acceptable methods in your protocol may require scientific justification and IACUC approval.

Example Alternatives Search:

For additional information on euthanasia of cats, including the impact of euthanasia agents on tissues, please refer to the examples of "alternatives" searches provided.

Federal Laws, Regulations, Policies:

1. Animal Welfare Act, as Amended (7 USC, 2131-2156).
2. Animal Welfare Act Regulations and Standards, Code of Federal Regulations, Title 9, Part 2-Regulations, Sections 2.31-2.33.
3. Health Research Extension Act of 1985, Public Law 99-158, November 20, 1985, "Animals in Research".
4. Public Health Service Policy on Humane Care and Use of Laboratory Animals, Revised September, 1986, Reprinted March, 1996.
5. USDA Animal and Plant Health Inspection Animal Care Policy Manual.
   Policy #11 - Painful/Distressful Procedures.
   Policy #12 - Written Narrative for Alternatives to Painful Procedures.
6. U.S. Government Principles For The Utilization And Care Of Vertebrate Animals Used In Testing, Research, And Training, Interagency Research Animal Committee.

Guidelines:

1. Guide for the Care and Use of Laboratory Animals, National Research Council, 1998.
2. National Center for Infectious Diseases, Center for Disease Control and Prevention (CDC).
3. Occupational Health and Safety in the Care and Use of Research Animals, National Research Council, 1997.

Texts:

1. Flecknell, Laboratory Animal Anesthesia, 2nd Edn., Academic Press, 1996.
2. Fox, Cohen, and Lowe, Laboratory Animal Medicine, Academic Press, 1984.
3. Hawk and Leary, Formulary for Laboratory Animals. 2nd Edn., Iowa State University Press, 1999.
4. Lewis, Morris, Jr., Hand. Small Animal Clinical Nutrition III. Publ: Mark Morris Associates, Topeka, KS. 1987.
5. Martin. The Laboratory Cat. CRC Press, 1998.
6. Wolfensohn and Lloyd, Handbook of Laboratory Animal Management and Welfare. 2nd Edn., Blackwell Science. 1998.