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be used for carrying or for lifting dogs from the oor. Dogs in trolleys, carts or containers should be made as comfortable as possible in order to minimize stress. Suitable substrate should always be provided.

15.4 Effects of transport There are few published data on the effects of transport on dogs (but see Leadon & Mullins 1991). Probably the greatest effect of transport on animals is psychological, in terms of fear and anxiety as a result of novel sensations and situations, and these are dif cult to quantify. More information is required in this area. Transporting animals leads to a temporary disturbance of normal physiology (Knowles & Warriss 2000). Much depends on the type of animal, the degree of adaptation prior to transport, and the circumstances of the journey. Of course, there will also be individual variation in response to transport. A suitable acclimatization period is essential, not only for animals to overcome any stress imposed by transport, but, more importantly, to allow suf cient time and opportunity for them to become accustomed to a new physical and social environment, and to allow a period of adjustment for physiological variables to return to normal before scienti c investigations are carried out. It is recommended that dogs are given an acclimatization period of at least 7 days following transport involving a journey in a vehicle and between sites, and at least 3 days when moved in individual containers between buildings on-site. Data on the effects of transport of dogs or other species moved within a building are scant. However, at least 24 h should be permitted for acclimatization where there is a permanent change of pen location, assuming a similar husbandry and care regime exists. At least 14 days of acclimatization may be needed if dogs are to be trained prior to the start of procedures (see Section 16).
Recommendations: Dogs should be given an acclimatization period of at least 7 days following transport involving a journey in a vehicle and
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between sites, and at least 3 days when moved in individual containers between buildings on-site. At least 24 h should be permitted for acclimatization following a permanent change of pen location, assuming a similar husbandry and care regime exists. At least 14 days should be allowed for acclimatization where dogs are to be trained prior to the start of procedures.

16

Handling and restraint

One of the most important ways to minimize stress for dogs in the laboratory is to ensure that they react well to handling. The dog should feel at ease when being approached, picked up, carried and restrained. All dogs should, therefore, experience adequate socialization with humans during the primary socialization period, combined with appropriate habituation and training related to their activities during studies (see Section 13). Positive interactions with humans, through regular and appropriate handling, should continue throughout the dogs lives. In handling and restraining dogs, it is essential to understand species-typical behaviour patterns and communication systems and to be able to accurately interpret signals and respond appropriately (see Section 4.3). It is important that all members of staff adopt a similar approach to handling and restraint and that interaction with the dogs is as consistent as possible. It is now becoming accepted that there should also be close liaison between breeders and users of dogs to ensure that handling methods are standardized between facilities. Breeds of dog vary in size, and techniques for handling, therefore, need adaptation. The con dence of the dog should initially be assessed by interacting with him/her in a passive and non-threatening manner. Squatting down to approach from the dogs level can help keep the dog calm and talking to him/her in a quiet but con dent manner can also be bene cial. Moments taken to speak to and stroke the dog may be repaid through reduction in the dogs anxiety and physiological variability (Wol e 1990).
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BVAAWF/FRAME/RSPCA/UFAW Joint Working Group on Re nement

Fig 14 When carried, dogs should be safely restrained and have their body weight supported

Fig 15 Dogs should be trained to tolerate restraint with positive reinforcement techniques

Direct eye contact and potentially threatening body postures must be avoided. When dogs are carried, they should be safely restrained and their body weight should be supported (Fig 14: Animal Welfare Institute); scruf ng should not be used for carrying or for lifting dogs from the oor or tables. In the procedure room, staff demeanour should be calm, con dent and quiet. Dogs can be stroked while on the examination table but care should be taken not to unintentionally reinforce fearful responses in this way. Temporary restraint is known to be potentially stressful to dogs (Knol 1989) but habituation and training can reduce the associated stress. If restraint is necessary to control a dog during a scienti c procedure, then the method used should provide the least restraint required to allow the procedure to be performed properly (Fig 15: Animal Welfare Institute). It should protect both dogs and personnel from harm and should avoid causing distress or unnecessary discomfort (National Research Council 1994). The duration of restraint should always be minimized. Minor procedures, such as taking a rectal temperature or administering a subcutaneous injection can usually be accomplished by using minimal restraint. During venepuncture, suf cient restraint and accuracy should be used to avoid repeated needle insertions and to prevent the development of painful haematomas.
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Where restraint or con nement is required as part of a scienti c programme in the UK, speci c justi cation must be included in the project licence, and details must be provided of the measures to be adopted to maintain high welfare standards, for example details of additional humananimal socialization and training for procedures (Animal Procedures Committee 1999). It is always preferable to train laboratory animals to cooperate during procedures and husbandry, rather than to automatically physically restrain them, unless there is likelihood that they will injure themselves or staff. Such training helps reduce the stress associated with procedures and is an essential part of everyday husbandry. Dogs should be trained to tolerate restraint, especially if they are to be restrained frequently or for prolonged periods. Positive reinforcement training should be used in the form of attention or praise, providing a favourite toy or morsel of highly palatable food (see Section 13). Positive reinforcement techniques can also be used to train laboratory dogs to accept procedures such as intramuscular injection, intravenous injection and oral dosing. Acclimatizing animals to the facility before procedures begin is likely to reduce distress and help build a positive relationship with animal care staff. Allow extra time for habituation and training animals to accept procedure rooms, restraint devices
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such as slings, metabolism cages (see Section 17.3), and stressful dosing procedures such as inhalation (by habituation to the mask) and oral gavage (by sham dosing for a xed period).

Dogs restrained in slings should be attended to at all times.

17

Procedures

Slings: Body slings are used to support dogs when it is deemed necessary to restrain them for prolonged periods (e.g. in inhalation studies). Introduction to body slings can be usefully incorporated into the initial habituation process at the breeding establishments, but speci c habituation to the sensation of being suspended by these devices should also be provided prior to individual studies. Quiet, calm dogs should be selected for use in procedures using slings. Dogs restrained in slings should be attended to at all times.
Recommendations: All staff with responsibility for handling dogs should be trained and competent in the appropriate handling techniques. There should be close liaison between breeders and users of dogs to ensure that handling methods are standardized. If restraint is necessary to control a dog during a scienti c procedure, then the method used should provide the least restraint required to allow the procedure to be performed properly. The duration of restraint should always be minimized. Positive reinforcement techniques should be used to train dogs to tolerate restraint. Socializing, habituating and training dogs so that they become used to humans and experimental and clinical procedures should be considered an essential element of everyday routines. Extra time should be allocated before procedures begin for training animals to accept procedure rooms, metabolism cages, restraint devices and stressful procedures. Where necessary, all dogs should be habituated and trained to stand quietly in slings. Any animal that does not settle quickly with initial training should not be used.
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Experimental procedures conducted in dogs can be divided into various categories: administration of substances; removal of body uids; metabolism cages; telemetry; anaesthesia, analgesia and perioperative care; and euthanasia. Detailed reports on most of these issues are available, and so only brief attention is given here.

17.1 Administration of substances It is a common requirement to administer substances to dogs as part of an experimental procedure, particularly for the purpose of testing the substance to be administered. Advice on best practice and re nement of administration of substances for various species, including dogs, can be found in the fourth report of the BVAAWF/FRAME/ RSPCA/UFAW Joint Working Group on Re nement (Morton et al. 2001). In addition, a joint European Federation of Pharmaceutical Industries and Associations (EFPIA)/European Centre for the Validation of Alternative Methods (ECVAM) working group has also recently published a paper addressing good practice on administration of substances by various routes for laboratory animal species, including dogs (Diehl et al. 2001). Commonly the route of administration is dictated by the expected clinical route of administration of the substance in question. Where there is an element of choice, the least invasive route with the least potentially adverse effects should be chosen. Such a choice may involve a degree of compromise between the potential for short-term and long-term effects. Therefore the sum of all of the procedures to be performed should be assessed in the development of the least aversive study design.
Recommendation: The administration of substances should be by the least invasive route appropriate,
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