least restriction necessary to achieve the scientific objective while maintaining animal well-being.

The development of animal protocols that involve the use of food or fluid regulation requires the evaluation of three factors: the necessary level of regulation, potential adverse consequences of regulation, and methods for assessing the health and well-being of the animals (NRC 2003b). In addition, the following factors influence the amount of food or fluid restriction that can be safely used in a specific protocol: the species, strain, or stock, gender, and age of the animals; thermoregulatory demand; type of housing; time of feeding, nutritive value, and fiber content of the diet (Heiderstadt et al. 2000; Rowland 2007); and prior experimental manipulation. The degree of food or fluid restriction necessary for consistent behavioral performance is influenced by the difficulty of the task, the individual animal, the motivation required of the animal, and the effectiveness of animal training for a specific protocol-related task.

The animals should be closely monitored to ensure that food and fluid intake meets their nutritional needs (Toth and Gardiner 2000). Body weights should be recorded at least weekly and more often for animals requiring greater restrictions (NRC 2003b). Written records should be maintained for each animal to document daily food and fluid consumption, hydration status, and any behavioral and clinical changes used as criteria for temporary or permanent removal of an animal from a protocol (Morton 2000; NRC 2003b). In the case of conditioned-response research protocols, use of a highly preferred food or fluid as positive reinforcement, instead of restriction, is recommended. Caloric restriction, as a husbandry technique and means of weight control, is discussed in Chapter 3.


Use of Non-Pharmaceutical-Grade Chemicals and Other Substances The use of pharmaceutical-grade chemicals and other substances ensures that toxic or unwanted side effects are not introduced into studies conducted with experimental animals. They should therefore be used, when available, for all animal-related procedures (USDA 1997b). The use of non-pharmaceutical-grade chemicals or substances should be described and justified in the animal use protocol and be approved by the IACUC (Wolff et al. 2003); for example, the use of a non-pharmaceutical-grade chemical or substance may be necessary to meet the scientific goals of a project or when a veterinary or human pharmaceutical-grade product is unavailable. In such instances, consideration should be given to the grade, purity, sterility, pH, pyrogenicity, osmolality, stability, site and route of administration, formulation, compatibility, and pharmacokinetics of the chemical or substance to be administered, as well as animal welfare and scientific issues relating to its use (NIH 2008).



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