Blood collection & injections in rodents and rabbit
Blood collection techniques in swine
Copyright 2006, University of Minnesota Board of Regents.
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Restraint and Handling of Animals
The use of proper restraint and handling techniques reduces stress to animals and also to the researcher. Handling stress represents an experimental variable and should be minimized whenever possible. Animals can inflict serious injuries to humans and to themselves as a result of improper handling.
- Animals experience stress as a result of shipping. All large animals must be allowed to acclimate to the facility for three days. During this time they may not be experimentally manipulated. Acclimation periods of up to one week are recommended for all animals.
- If a study will involve significant handling of animals it is recommended that the animals be acclimated to the handling. Prior to experimental manipulation, handle the animal on a regular basis in a non-threatening situation, e.g. weighing, petting, giving food treats. Most animals, even rodents will respond positively to handling and will learn to recognize individuals.
- Handle animals gently. Do not make loud noises or sudden movements that may startle them.
- Handle animals firmly. The animal will struggle more if it sees a chance to escape.
- Use an assistant whenever possible.
- Use restraint devices to assist when appropriate.
- Chemical restraint should be considered for any prolonged or potentially painful procedure.
The methods described below will assist with performing basic manipulations. Alternate techniques may be needed for special procedures. Most of these methods are also demonstrated in video tapes available to investigator. For other information on animal handling or for individual training, contact RAR at 624-9100. An excellent website containing laboratory biomethodology for rodents and rabbits is also available with descriptions and pictures of drug administration, blood collection and sex determination.
Needle Re-Use Policy
The use of a new sterile needle and syringe for each animal when giving parenteral injections (intraperitoneal, subcutaneous, intravenous, intramuscular, etc.) is the recommended best practice to prevent the horizontal transfer of contamination between animals. However, the IACUC recognizes that there are some instances where it may be justified to use the same needle and syringe for multiple animals, usually in rodents. In those instances the Principal Investigator must provide justification to the IACUC and must adhere to the following guidelines. Use of the same needle and syringe may be permitted with justification on animals housed in the same cage. The needle must be assessed for continued sharpness and the presence of barbing or burring of the tip between animals. If dullness or needle deterioration is found, a new needle must be used.
Tail restraint, as described below is adequate for examining animals and transfering them to another cage.
These methods may be used to perform minor, non-painful procedures such as injections or ear tagging.
RATS may be handled by the tail, with precautions similar to those used for mice, with emphasis on only grasping the tail base. Holding the tail distal to the base can result in a de-gloving injury to the tail that will require surgical repair or euthanasia.
This method should be used to restrain a rat for injections and other minor procedures.
Because hamsters do not have tails, they must be grasped firmly by the loose skin of its back, or handled in a manner similar to the rat.
GUINEA PIGS rarely bite, but are very easily frightened and will vocalize and squirm to avoid restraint. The hind limbs must be supported at all times to prevent the animal from injuring its back.
RABBITS are very susceptible to lumbar spinal luxation, resulting in paralysis. It is necessary to support the animal's hindquarter at all times. Although rabbits seldom bite, they can inflict painful scratches with their hind legs. One way of lifting a rabbit is by grasping the skin over the shoulder with one hand and gently lifting it with the other arm cradling the body, the head nestled in the crook of your arm. Rabbits must never by lifted by the ears.
CATS are often cooperative enough to be restrained on a table by the loose skin at the back of the neck and hips, or with one hand restraining the body and the other restraining the head. A fractious cat may have to be wrapped in a heavy towel for restraint with any needed limbs carefully withdrawn for treatment.
A slip lead is highly recommended for working with dogs. A dog should always be carried with proper support. The dog can be restrained in lateral recumbancy or in a sitting position for injections and minor procedures. For venipuncture, the handler can restrain the dog on a table with one arm around its neck. The other hand is then free to restrain the body if necessary or to occlude the vein for the person with the syringe. A shy or fearful dog may need extra time spent with it to make it more comfortable. Moving slowly and speaking quietly will help to prevent alarming the animal.
An intractable dog may need to be muzzled. A commercial muzzle may be purchased, or a gauze muzzle may used as described below.
Pills are easily administered to most dogs if the proper technique is used.
NONHUMAN PRIMATES, no matter how small, can be dangerous. Chemical immobilization with ketamine is nolrmally used. Injections can be given to a confined animal with the help of a squeeze cage.
Physical restraint of a conscious animal should only be attempted by trained, experienced personnel. Animals may be pole and collar trained if they will be handled frequently. Tether systems are recommended if animals must be administered drugs or if blood must be collected frequently.
Absolute requirements for handling of nonhuman primates include attending a training module given by RAR (contact 624-9100 to schedule), and wearing appropriate protective clothing. In addition, nonhuman primate users should be familiar with procedures to follow in case of a bite or scratch and the location of bite kits.
If a nonhuman primate has escaped, close all doors and contact RAR at 624-9100. The animal may be recaptured using a net or a dart gun.
GOATS, SHEEP and CALVES
- Restrain against a wall or in a corner by placing a knee firmly in the flank.
- Restrain for blood collection by backing the animal into a corner and straddling them at the shoulder and firmly restraining the head and neck.
- Use a halter over their head and face.
- A sheep can be held for bleeding, shearing or hoof trimming by sitting the animal up on its hind end, leaning back against the restrainer.
- For long term restraint of sheep in the laboratory, a canvas sling and rack is available from several commercial suppliers. Animals are easily acclimated to such slings, and can be comfortable and relaxed enough to fall asleep in them.
- Additional references on handling of agricultural animals is available from the USDA.
- Temple Grandin's Website on Low-Stress Handling of Farm Animals
RESTRAINT AND HANDLING OF SWINE
By Dr. Jack Risdahl
Additional information on restraint of, and blood collection from, swine may be found on the UCDavis website. http://ehs.ucdavis.edu/animal/vet_care/training/Pig.cfm
Pigs in general are friendly and docile but will react severely to poor handling or a stressful environment. Pigs can be very vocal. If pigs are chronically stressed they will become skittish and fearful. Handling and restraint in pigs relies greatly on treating the pigs in a humane manner. The benefits of treating pigs well include reducing apprehension, fear and stress in the pigs. There are several levels of restraint and handling, from touching and coaxing a pig to restraining a pig for chronic procedures.
Touch is a very important aid to good husbandry.
When approaching a pig be sure it is made aware of your presence. If pigs are startled they may cause injury to themselves or others in the pen. The best way to make pigs aware of your presence is to use your voice. It is important to use a soft soothing voice and not angry, loud, high pitched tone of voice which might startle or stress the animal. Pigs quickly learn to recognize voices, especially if they are associated with food. As pigs become familiar with handlers, the sound of a familiar voice is often calming to the animal. It is important to use touch when developing a rapport with pigs. This applies especially to the researcher who must collect frequent samples or data from pigs. As with voice, gentle petting and hand contact should be associated with feeding time or treats and the pig will become aware of the person in the vicinity and become adjusted to that persons presence. Probably one of the best forms of restraint in pigs is the use of food. Pigs are highly oriented to food and if they are comfortable with the handler will most often stand and eat while minor procedures and examinations are being performed on them. One can often flush catheters, give injections, treat minor wounds and take temperatures while pigs eat. The use of all three procedures - voice, touch, and food, will be the best investment in reducing stress among research swine and will ultimately reward the researcher with a happy stress free subject.
The giving of food is one of the most effective forms of basic restraint in the pig.
Picking Up Pigs
Pigs best tolerate being picked up in a "horizontal" fashion oriented to the ground. Pigs should not be picked up by the legs or held upside down as this will stress the animal and you will loose their trust. Usually only smaller animals may be picked up while larger animals (>35-40 kg) must be moved by alternative means. Smaller pigs may be easily picked up with their body supported while their legs hang. To perform the procedure in larger pigs place one arm under the chest cranial to the thoracic limbs and the other arm cranial to the pelvic limbs under the abdomen picking up the pig in a "scooping" fashion. Alternatively the arm may be placed caudally just above the pig's hock, hence supporting the animal by the pelvis rather than the abdomen. All handlers must beware to lift with legs and not back as injury can easily result - pigs are usually heavier than they appear! Always avoid picking pigs up by one leg or by the ears as injury may result!
The small board used to apply pressure to the side of a pig.
Pigs are best moved in a metal (box style) transport designed for use with large animals. At times this is not possible and pigs must be walked to their destination. When moving a pig always remember pigs will move away from walls toward openings. This is an advantage since one can use a "hog board" to simulate walls. The board is fashioned with a handle so that one can place it to the side, rear or front of the pig to direct them. Excessive force should not be needed to move a pig and is mostly counterproductive as pigs will become excited and belligerent. It should be remembered pigs will refuse to move if the place you wish them to go is dark (e.g. from daylight into a dark room). Sometimes pigs may be coaxed with food along with the use of the board. When pigs are unruly and where control is needed, pigs may be tethered in a harness and controlled by "holder" so that the pig does not run away. Often the use of the hog board may be used to stop pig and slow them down if they are moving too rapidly. The board may also be used to restrain a pig in a corner while minor procedures are performed. The size of the board varies depending on the size of pigs used and application. In general if the board is at least as tall as the pig and 2/3 to about as long as the pig it will usually suffice.
Several designs for slings to restrain pigs have been described. The most commonly used is that described by Panepinto et al 1983. Here the pig is placed in a hammock with four holes for the limbs. The hammock is supported by a metal frame. These are available in free standing or winch styles (so larger pigs may be raised by winch). The pig is placed in ventral recumbency in the sling with its limbs tied loosely to the frame. It has been our experience that this form of restraint requires some degree of training for pigs to acclimate to. In general most pigs will become stressed the first several times they are placed in the sling. Positive reinforcement (treats, petting) and repetition usually calms them down so that they may be restrained for extended periods in the sling. We have generally used a training period of two weeks prior to experimental procedures with a minimum of 30 min./day in the sling. In our experience one or two hours is about the most a pig will tolerate.
Acclimation and Socialization
It should be remembered that pigs are social animals and have a rigid dominance hierarchy. If animals are group housed they will generally fight to establish dominance for the first 24-48 hours. Dominance in pigs is almost directly related to size. The largest animals are dominant and smallest are submissive. Be sure to match weights as close as possible when introducing new pigs to each other. Smaller pigs may be injured by larger pigs. Be sure to monitor pigs for the introduction period so that they do not cause major injury to each other - they will fight. Always remember that newly arrived pigs are stressed from transport. Do not initiate experimental procedures in the first few days of arrival. This is just common sense as immune function and physiologic parameters are often altered by stress. We like to see an acclimation period of two weeks so that pigs may adapt to their new environment and establish rapport with handlers.
- A butterfly needle can be attached to a syringe for administering injections to swine, allowing them to move during the injection without displacing the needle.
- Intravenous injections may be given in the ear veins.
- Oral drugs may be administered ground or whole mixed with a food treat.
- Pills can be administered orally as with dogs. However, the handler must be sure to get the pill all the way behind the tongue and must avoid being bitten.
- Some drugs may be administered rectally. A literature review should be performed for the drug in question prior to attempting this.
- Indwelling central venous catheters are recommended if animals will be receiving drugs on a regular basis.
[Link to blood collection techniques in swine]
Restraint devices such as rabbit or rodent restrainers, swine slings or monkey chairs are useful for certain non-painful procedures. However, certain guidelines should be followed when using these devices.
- Animals should be adapted to the restraint devices. This means that for long-term restraint (i.e. more than an hour), it is advisable to "train" the animal to the device by placing it into the device for successively longer intervals until the maximum time of restraint can be achieved without causing distress to the animal.
- Animals in a restraint device be regularly monitored. This means not leaving the area for long intervals unless someone else is available to monitor the animal. Animals have an uncanny ability to attempt escape from devices, if they don't succeed completely, they may end up with a limb or their head entrapped. This could result in ischemia or hypoxia.
- Animals should have access to food or water at appropriate intervals, even when restrained, unless doing so would interfere with the goals of the experiment. Food or water should be offered twice daily. For rabbits and rodents, water should be offered more frequently.
- Animals should be released from restraint devices at least daily and allowed unrestrained activity to prevent muscle atrophy and skin necrosis, unless this interferes with achieving the experimental goals and is documented in an approved IACUC protocol.
Typical rodent restrainers
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The information contained in this site is intended as a reference for University of Minnesota investigators, and animal husbandry and veterinary staff. Drug information and dosages are derived from a variety of sources and do not necessarily guarantee safety or efficacy. Information obtained through this site should not be relied upon as professional veterinary advice. Any medications administered or procedures performed on animals should only be performed by or under order of a qualified, licensed veterinarian.