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J Husbandry This chapter provides guidelines for the care of laboratory dogs. The first section, on housing, details design and construction considerations for facilities that house dogs, as well as for primary enclosures (here defined as cages and pensJ. The subsection on facilities contains information on build- ings, rooms, and outside areas for containment of dogs, and that on environ- ment and environmental control describes mechanisms for controlling the environment and gives the legislatively mandated ranges for temperature, humidity, and ventilation. The remaining information in this chapter is supplemented by discus- sions in other parts of this report. For example, Chapter 4 ("Management of Breeding Colonies") contains sections on food for puppies and gestational or lactating dams and on record-keeping for a breeding colony that amplify the sections on food and identification and records in this chapter. Social- ization of puppies is also discussed in Chapter 4. Modified primary enclo- sures and bedding for dogs with specific disorders are described in Chapter 6 ("Special Considerations". The 1985 amendment to the Animal Welfare Act required the U.S. Department of Agriculture (USDA) to establish standards for exercise for laboratory dogs, and they were established in 1991. A federal court has now found that the regulations concerning exercise for dogs are inadequate and ordered that new regulations be written. This committee has reviewed the available information relevant to exercise, space, and well-being of dogs, 11
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2 DOGS: LABORATORY ANIMAL MANAGEMENT and it has found that, as was the case in 1985, it is inadequate to formulate objective standards. Although knowledge of canine behavior is leading to a consensus that opportunities for social interaction with people, other dogs, or both are important for promoting canine well-being, no similar consensus is avail- able concerning fitness and exercise. Another issue is the notion that a single standard can provide optimal care for all dogs. It is generally recog- nized that such factors as breed, physical conformation, age, health status, past experiences, and general behavioral characteristics influence what con- stitutes adequate space and exercise. For example, a dog undergoing a surgical procedure might require a restricted space to limit its activity. Once the dog has recovered from the surgical procedure, a different space and exercise regimen can be implemented. Likewise, the space and type and duration of exercise required for Alaskan sled dogs in working condition is quite different from that required for Shih Tzu and other brachycephalic breeds. Finally, medical research benefits from the availability of dogs with inherited disorders similar to those of humans, and the presence of these disorders in dogs imposes the same types of restrictions that human patients must endure. Unsupervised exercise is often contraindicated in dogs with heart and metabolic diseases. Similarly, the construction and layout of primary enclosures for dogs with such conditions as muscular dystrophy, bleeding disorders, blindness, or Ehlers-Danlos syndrome must be carefully considered to avoid compromising their health and well-being. The most important objective for those responsible for housing dogs should be to achieve an overall high level of care, rather than to conform rigidly to specific standards. Animal well-being must be assessed case by case by those qualified to do so. The regular evaluation of animal well- being is an important aspect of any husbandry and animal-care program and serves as a measure of the appropriateness of animal-care procedures. Pro- cedures that are ultimately linked to the well-being of the individual are defined as performance standards. The committee strongly recommends that performance standards, coupled with sound professional judgment, be used to develop space requirements and exercise programs for dogs. This committee is firmly convinced that performance standards are ultimately better for each dog's physical and behavioral well-being than engineering standards, which might lack the flexiblity necessary to meet the needs of all dogs. HOUSING Facilities Housing facilities for dogs must be designed and constructed so that they are structurally sound, protect animals from injury, contain animals
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HUSBANDRY 13 securely, and prevent entry of other animals (9 CFR 3.1a). Dog facilities vary in size and complexity, depending on their purpose (e.g., holding or breeding), colony size and type (e.g., specific-pathogen-free or conventional), and breed. The design of breeding facilities should address the following: · The design should facilitate the conduct of research. · There should be sufficient space for expansion, both for adding an mats and for increasing ancillary operations. . Breeding facilities should have sufficient space to house dams with litters and the progeny. The design should promote effective sanitation and husbandry proce dures. · Operation of the facility should be efficient and cost-effective. · Construction should be economical. The physical facilities and equipment should be constructed and oper- ated to fulfill the following criteria: · Contamination from areas adjacent to, but not part of, the facility should be minimized. The locations of equipment washing and sterilizing, food and bedding storage, quarantine, treatment, receiving and shipping, shipping-crate storage, mechanical services, shops, offices, and laboratories should minimize crossovers from soiled or contaminated to clean areas. Clean material and equipment should not come into contact with soiled and contaminated material and equipment. . There should be sufficient control of temperature, humidity, ventila- tion, and lighting to provide the animals with appropriate conditions for their comfort and well-being. · Behavioral well-being should be considered by allowing for visual contact between dogs, social housing, exercise areas, and other appropriate areas. · The entry of vermin should be prevented. . Provisions should be made for lunchrooms, locker rooms, and toilets for animal-care personnel. . Caging equipment and feeding and watering devices should provide a safe environment, make food and water readily available, minimize the opportunity for transmission of diseases and parasites, and make sanitation and sterilization efficient. · Auxiliary equipment such as washing machines, cage racks, rolling equipment (e.g., dollies, tables, and carts), and fixed equipment (e.g., cabi- nets, sinks, and shelving) should be designed, fabricated, and used to pro- mote maximal sanitation and operating efficiency.
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4 DOGS: LABORATORY ANIMAL MANAGEMENT When a dog facility is designed to be part of a larger facility housing other species of animals or part of a multipurpose building with offices and research laboratories, the physical relationships between areas must be care- fully planned (NRC, 1985a). Those establishing operating procedures should use the best available information on physiology; nutrition; genetics; behav- ior; animal breeding, care, and maintenance; colony management (produc- tion and research); and disease control. Dogs can be housed in indoor facilities, outdoor facilities, or a combi- nation of the two (sheltered housing facilities). If the site is exclusively indoors, the only factors that influence site selection are local zoning regu- lations, the ability to control odors and noise, the availability of appropriate utilities (e.g., sewerage and water) (9 CFR 3.1d), and the proximity to other businesses. Indoor facilities should be constructed and maintained in com- pliance with CFR, Title 9, Part 3.2 and the Guide (NRC, 1985a), as summa- rized below. Indoor Facilities Walls. Exterior walls should be fire-resistant and impervious to ver- min. To facilitate cleaning, interior walls should be smooth, hard, and without pits or cracks, and they should be capable of withstanding the impact of water under high pressure and scrubbing with cleaning agents (e.g., detergents) and sanitizing agents (e.g., disinfectants). They should be protected from damage caused by movable equipment. Ceilings. Ceilings should be smooth, moistureproof, and free of imper- fect junctions. Surface materials should be capable of withstanding scrub- bing with detergents and disinfectants. undesirable. Exposed pipes and fixtures are Floors. Floors can be constructed of a variety of materials that are smooth, moistureproof, nonabsorbent, and skidproof; that are resistant to wear and the adverse effects of detergents, disinfectants, acid, and solvents; and that are able to support heavy equipment without being gouged, cracked, or pitted. They should also be easy to clean. Drainage. Drainage must be adequate to allow rapid removal of water (9 CFR 3 . 1 f). _ --So ~ 1 If floor drains are used, they should be constructed and maintained in accordance with the Guide (NRC, 1985a). Rim flush drains should be at least 6 in (15.2 cm) in diameter. Porous trap buckets installed in the drains aid in cleaning and screen out solid waste. Floor drains must contain traps that prevent backflow of sewage and gases (9 CFR 3.1f). If
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HUSBANDRY 15 unused floor drains are present, they should be closed with gaslight seals that are flush with the floor surface. Doors. All rooms should have doors. External doors should have adequate latches and locks and should be verminproof when closed. If they are left open during warm weather, adequate screening is essential. All door frames should be sealed to walls and partitions with caulking com- pound or a similar material. Ports in animal-room doors allow personnel to observe the dogs with- out entering the rooms, prevent injury to personnel while they are opening doors, and provide a way to verify that room lights are on at appropriate times. Experience has shown that doors at least 42 in (107 cm) wide and 84 in (213 cm) high allow free passage of cages and equipment. The doors should be equipped with locks and kickplates and should be self-closing. Outside windows. Outside windows and skylights might not be desir- able, because they can contribute to unacceptable variations in temperature and photoperiod. Other problems associated with outside windows and skylights include dust and bacteria buildup on frames; drafts, and increased ventilation costs. Washrooms and sinks. Washing facilities for personnel (e.g., basins, sinks, or showers) must be provided and must be readily accessible (9 CFR 3.1g). Sheltered Housing Facilities A sheltered housing facility, as defined by the Animal Welfare Regula- tions (AWRs), is a facility that provides shelter, protection from the ele- ments, and protection from temperature extremes at all times (9 CFR 1.13. It can consist of runs or pens in a totally enclosed building or indoor- outdoor runs with the indoor runs in a totally enclosed building. The re- quirements for the sheltered portion of such facilities are identical with those for indoor facilities, with the additional stipulation that the shelter structure must be large enough to permit each animal to sit, stand, and lie down in a normal manner and to turn around freely (9 CFR 3.3~. The outdoor portion of a sheltered housing facility should be constructed to prevent the introduction of vermin. Outdoor floor areas in contact with animals should be constructed of hard, moisture-resistant material and be properly drained. The use of compacted earth, sand, gravel, or grass is discouraged. The sides of runs can be constructed of chain-link fencing and steel posts or pipe frames or, when necessary to prevent fighting or injury, of solid concrete block coated with sealant. Fencing at the lower ends of
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6 DOGS: LABORATORY ANIMAL MANAGEMENT runs and pens should be high enough above the surface to permit adequate drainage but not high enough to allow young puppies to escape. Curbs at least 6 in (15.2 cm) high should be constructed between runs to help pre- vent the spread of microorganisms during washing. Curbs 24-30 in (61.0- 76.2 cm) high might be necessary in runs in which the dog population is constantly changing. Higher curbs might be beneficial in whelping-pen runs to reduce the anxiety of nursing bitches. Run doors or gates should have well-made latches that can be easily opened by animal-care personnel but not by the dogs. Special consideration must be given to removing animal wastes and controlling noise. Outdoor Facilities The AWRs, with some restrictions, permit facilities to house dogs solely outdoors, provided that each animal has access to a structure (consisting of a roof, four sides, and a floor) that furnishes adequate protection from cold, heat, the direct rays of the sun, and the direct effects of wind, rain, and snow (9 CFR 3.4~. In general, this type of housing is discouraged for dogs being used in an experimental protocol, because environmental factors, in- fectious agents, and vermin are difficult to control. In other instances (e.g., in protocols requiring acclimation or in breeding colonies maintained in temperate climates), outdoor facilities might be adequate. Environment and Environmental Control An important part of maintaining the health and well-being of labora- tory animals is control of the environment. In nature, animals respond to environmental changes both behaviorally and physiologically in a manner that will maintain homeostasis. In an animal room, a behavioral response might not be possible, and the animal must deal with an altered environ- ment physiologically. Therefore, it is necessary to control the environment to avoid physiologic changes. Besch (1985) has reviewed environmental factors that can effect the biologic responses of laboratory animals. Temperature and Humidity Temperature and humidity are important considerations in a dog facility (Beech, 1985~. Dogs can tolerate moderate ranges of temperature and weather, provided that they have appropriate amounts of food and water, have access to shelter, and are allowed sufficient time to acclimate to their environment. The Guide recommends that room temperature for dogs be maintained within a range of 18-29°C (64.4-84.2°F) and relative humidity within a range of 30-70 percent. The AWRs require that the ambient temperature in indoor
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HUSBANDRY 17 facilities not fall below 7.2°C (45°F) or rise above 29.4°C (85°F) for more than 4 consecutive hours when dogs are present (9 CFR 3.2a). Except as approved by the attending veterinarian, ambient temperature must not fall below 10°C (50°F) for dogs not acclimated to lower temperatures, breeds that cannot tolerate lower temperatures, and young, old, sick, or infirm dogs (9 CFR 3.2a). Dogs recovering from general anesthesia are frequently hypothermic. Every attempt should be made to maintain normal body temperature during surgery and recovery. This can be accomplished by using supplemental sources of heat (e.g., heating pads and heat lamps), by avoiding direct contact with heat-conducting surfaces (e.g., metal), and by maintaining the postoperative recovery cage at 27-29°C (80.6-84.2°F) (NRC, 1985a). New- born pups have poorly developed thermoregulatory mechanisms and might require supplemental sources of heat. Temperatures of 29.4-32.2°C (85- 90°F) have been suggested for the first week of life (Poffenbarger et al., 1990~. Each room should be provided with temperature controls and high- and low-temperature alarms. Graphic recorders are useful for monitoring sys- tem performance. Ideally, the temperature controls should allow individual adjustments in dry-bulb temperature of + 1°C (+ 2°F) within the range of 18.3-29.4°C (65-85°F). Relative humidity should be maintained at 30-70 percent throughout the year (NRC, 1985a). It is important to control sources of humidity, such as cage-cleaning equipment, transient loads from cleaning water (Gorton and Besch, 1974), and thermal and mass loads from animals (Beech, 1991~. Low humidity can contribute to respiratory distress; and coughs, pneumonitis, and other problems can follow. High humidity impairs efficient body- cooling (Beech, 1991). Ventilation Ventilation serves multiple functions. It supplies oxygen; removes heat generated by animals, lights, and equipment; dilutes gaseous contaminants; and helps to control the effects of infiltration and exfiltration (slough and Gamble, 1976; Edwards et al., 1983~. Gorton et al. (1976) have reported a method for estimating laboratory animal heat loads. Indoor facilities must be sufficiently ventilated when dogs are present to provide for their comfort and well-being and to minimize odors, ammo- nia concentrations, drafts, and moisture condensation. Auxiliary ventilation must be provided when the ambient temperature is 29.5°C (85°F) or higher (9 CFR 3.2b). It is commonly thought that 10-15 volumetric changes per hour with outside air must be provided to animal rooms and that air must not be recirculated. As a consequence, animal facilities are generally venti
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18 DOGS: LABORATORY ANIMAL MANAGEMENT lated with "one-pass" air, although the Guide (NRC, 1985a) includes provi- sions for alternative methods of providing equal or more effective ventila- tion. Besch (1992) has reviewed alternative methods of ventilation. Ventilation system design and construction considerations include the following: Diffusers and exhaust openings should be located and controlled to prevent drafts. · Outside openings and exhaust-ventilation grillework should be screened to prevent entry of vermin. Screening should be cleaned regularly. · Air pressure in clean areas and animal rooms should be greater than that in public and refuse areas. Where pathogenic organisms are present, a negative-pressure system is necessary. · Ventilating mechanisms should be equipped with suitable alarm sys- tems that will be activated if the temperature moves outside the desired range or if power fails. . . Supplemental exhaust fans or exhaust systems increase drying and reduce humidity when fixed equipment is being washed. If such systems are used, they should be permanently mounted in external windows or wall openings, their frames should be sealed to the building structure, and the systems should be screened. · Emergency power sources should be available in case of power fail ure. Power and Lighting Electric systems should be safe, furnish appropriate lighting, and pro- vide a sufficient number of outlets. Lighting systems should allow for either manual or timer-controlled changes in illumination levels or photope- riods, and timer performance should be checked regularly. Lighting fix- tures, switches, and outlets should be sealed to prevent entry or harboring of vermin. Moistureproof switches and outlets should be installed where water is used in cleaning. Emergency power should be available. Illumination must be adequate and uniformly diffuse throughout each animal room to allow proper cleaning and housekeeping, to permit inspec- tion of animals, and to maintain the animals' well-being (9 CFR 3.2c). Light levels of 323 lx (30 ft-candles), measured 1.0 m (3.3 It) above the floor, appear to provide sufficient illumination for routine animal care (Bellhorn, 1980; NRC, 1985a). A regular diurnal lighting cycle must be provided (9 CFR 3.2c).
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HUSBANDRY Noise Control 19 Barking dogs can be a nuisance both to personnel working in animal facilities and to the adjacent population. Self-generated noise of 80-110 dl3 (Peterson, 1980; Sierens, 1976) has been measured in dog rooms. The effects of noise on animals are reviewed in the Guide (NRC, 1985a). Noise-control measures should be implemented in both indoor and out- door environments. Sound transmission can be reduced by using concrete to build walls, covering concrete walls with sound-attenuating material, and eliminating windows (NRC, 1985a). Pekrul (1991) has discussed other means of decreasing noise in animal facilities. Sound-attenuating materials may be bonded to walls or ceilings only if they can be sanitized and will not harbor vermin. Outdoor runs must be designed and constructed to comply with local noise ordinances. Chemicals and Toxic Substances Many of the chemicals used in animal facilities for cleaning, sanitizing, pest control, and other purposes can be toxic to housed animals and person- nel. In addition, some materials used in construction for coating surfaces can react with certain cleaning and sanitizing agents to produce toxic gases, including chlorine. Where possible, the use of chemicals should be avoided. For example, adequate ventilation is more effective than chemicals in elimi- nating most animal-room odors, provided that air inlets are not placed near the building exhaust. Newberne and Fox (1978) and Besch (1990) have reviewed chemicals and other toxicants found in animal facilities. Where chemical agents must be employed, it is essential to be familiar with their potential toxicity and to develop procedures for using and dispos- ing of them properly. Noxious chemicals should not be used to clean animal facilities. Adequate rinsing is essential to prevent the skin irritation or allergic reactions that can be caused by some cleaning and sanitizing agents (e.g., pine oil). Primary Enclosures Primary enclosures should facilitate research while maintaining the health and well-being of the dogs. They must confine dogs securely, enable them to remain clean and dry, protect them from injury, and contain sufficient ;; ~o allow Rem lo sit, ~e, stand, rum around, and walk normally (9 CFR 3 6a). The design should allow inspection of cage or pen occupants without disturbing them and provide easy access to feeding and watering devices for filling, changing, cleaning, and servicing. ~ ~ ~ _ ~ _ ~ 1 1 _ _ _ .1 ~1 · . ~
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20 DOGS: LABORATORY ANIMAL MANAGEMENT Cages or pens should be fabricated of smooth, moisture-impervious, corrosion-resistant materials that can be easily sanitized and sterilized. Floors must be constructed to preclude entrapping toes, dew claws, or collars. Expanded metal or plastic-covered metal mesh is satisfactory for pens or runs, provided that the dogs' feet cannot pass through the openings (9 CFR 3.6a2x). Pen floors must have adequate drainage. Each cage and pen should have a hinged or sliding door that covers the opening sufficiently to prevent escape of the occupants. Each door should have a latch that holds the door securely closed. Space Recommendations The AWRs require that the floor space for each dog equal at least the "mathematical square of the sum of the length of the dog in inches (mea- sured from the tip of its nose to the base of its tail) plus 6 inches t15.24 cm]," expressed in square feet (9 CFR 3.6cli,. In addition, the interior height of each enclosure must be "at least 6 inches t15.24 cm] higher than the head of the tallest dog in the enclosure when it is in a normal standing position" (9 CFR 3.6cliii). Each bitch with nursing pups must be given additional floor space based on breed and behavioral characteristics and in accordance with generally accepted husbandry practices, as determined by the attending veterinarian (9 CFR 3.6clii). The additional space for each nursing pup must be at least 5 percent of the minimum required for the bitch, unless otherwise approved by the attending veterinarian (9 CFR 3.6clii). Minimal space recommendations for dogs are also given in the Guide (NRC, 1985a, p. 14~. These requirements and recommendations are based prima- rily on professional judgment and convention. The few scientific studies on this subject have focused on how enclo- sure size affects movement, activity patterns, and physical fitness. Clark et al. (1991) found no decreases in physical fitness, as measured by heart rate and muscle enzyme (succinate dehydrogenase) activity, when dogs were housed in cages or runs of various sizes that complied with federal stan ~ - 1 _ _ _1 _ _ ~ ~ ~ ~ ~ ~ ~ ~ ~ _ _ . ~ ~ ~ . ~ V1_ A _ clarets and gulclellnes; However, modest Decreases in llr~less Wt;1~ l(JUllU WllC:11 dogs were housed in cages smaller than mandated by the AWRs. It has been shown that, in general, dogs are more active in pens and runs than in cages; however, dogs housed in the largest enclosures are not always the most active (Hefts et al., 1992; Hits et al., 1977; Hughes and Campbell, 1990; Hughes et al., 1989; Neamand et al., 1975~. Enclosure size has not been demonstrated to affect the musculoskeletal system (Newton, 1972), cortisol concentrations (Campbell et al., 1988; Clark et al., 1991), or se- lected measures of immune function (Campbell et al., 1988~. Although they provide interesting and relevant information, the studies do not provide
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HUSBANDRY 21 sufficient objective, scientific data on which to base space requirements for dogs. To set standards based on scientific data, one must show a correlation between cage size and behavioral well-being. That poses two problems: it is not clear how to define and measure behavioral well-being, and the deter- mination of well-being depends on human interpretations of the data. Move- ment and activity patterns are unlikely to be sensitive behavioral measures, because a dog's activity can be increased without improving its well-being (e.g., if there is locomotor stereotypy or increased activity caused by social isolation or competition for space). Moreover, the definition of movement varies between studies, so it is difficult to compare and interpret results. It is generally accepted that a variety of perspectives are needed to assess well-being, including measures of physical health, of neuroendocrine and immunologic responses to stress, of the ability to respond effectively to social and nonsocial environments, and of behavior. Scientific data on dogs are inadequate to support any such assessment relative to enclosure size. EXERCISE AND ENVIRONMENTAL ENRICHMENT The requirements for providing opportunities for dogs to exercise are specified in the AWRs (9 CFR 3.8~. The following paragraph summarizes the AWRs now in effect. It is incumbent on the reader to keep abreast of changes that might occur as the result of further federal court or USDA actions. Dogs over 12 weeks old, except bitches with litters, must be given the opportunity for regular exercise if they are kept individually in cages, pens, or runs that are less than 2 times the AWR-required floor space. Dogs housed in groups do not require exercise periods, provided that the total floor space of the cages, pens, or runs equals the sum of-the AWR-required spaces for the dogs if housed individually. If a dog is housed without sensory contact with other dogs, it must receive positive physical contact with humans at least once a day. Forced-exercise programs (e.g., swim- ming or walking on treadmills or carousel devices) are not considered to comply with the AWRs. Each institution is responsible for developing a plan for providing exercise. The plan must be approved by the attending veterinarian and must be made available to USDA on request. Exceptions to the requirement for exercise can be made by the attending veterinarian case by case or, if exercise is inappropriate for a scat ntific protocol, by the institutional animal care and use committee (IACLJC) In the former in- stance, the exemption from exercise must be reviewed every 30 days, unless it was granted because of a permanent condition (9 CFR 3.8d). In the latter instance, exemptions must be reviewed at appropriate intervals, as deter- mined by the IACUC, but not less often than every 6 months (9 CFR 2.31)
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24 DOGS: LABORATORY ANIMAL MANAGEMENT consulted by those designing exercise and social interaction plans or when other questions arise concerning the behavioral well-being of dogs. FOOD Selecting Optimal Rations Many commercially available dog foods contain all essential nutrients in their required proportions, as outlined in Nutrient Requirements of Dogs (NRC, 1985b) and the Association of American Feed Control Officials' Official Publication 1993 (AAFCO, 1993~. These foods are manufactured in dry, semimoist, and canned forms. Dogs should be fed only complete and balanced diets. Specific procedures should be followed to ensure that stored foods do not become deficient in nutrients (NRC, 1985a). Diet quality can be evaluated by examining the label for a statement of nutritional adequacy, which must be present on all dog-food products sold across state lines. This statement informs the purchaser whether the prod- uct has been approved for use as a complete ration for specified life stages (i.e., growth, maintenance, or pregnancy and lactation). Approval is ob- tained by one of the following means: · Each of the diet's individual ingredients is analyzed for all essential nutrients; the sum of these nutrients in all ingredients must meet or exceed the nutritional requirements of the animal for specified life stages. . The product itself is chemically analyzed and shown to meet or ex ceed the essential-nutrient requirements for specified life stages. . The product passes a feeding trial as specified by the Association of American Feed Control Officials. If the product fails to be approved, it must be labeled for use as a dietary supplement only and is not appropriate for use as a dog food. Of the three means of approval, only the feeding trial evaluates the availability of the nutrients in the product. Dog foods approved by that method should be used whenever possible. If such a diet cannot be used, because it would interfere with the experimental design (e.g., nutritional studies with purified diets), the manufacturer of the diet to be used should be consulted about experience with the diet's performance under given conditions. Many commercially available dog foods, although designed for a speci- fied life stage, are approved and adequate for use during all life stages. Most growth formulations will meet the requirements for gestation, lacta- tion, and maintenance. Similarly, most gestation-lactation products also meet requirements for growth and maintenance. Some foods intended for maintenance will meet the criteria for more than one life stage. However,
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HUSBANDRY 25 no food should be used for growth, gestation, and lactation unless its label states that it meets or exceeds nutrient requirements for these life stages. Special therapeutic diets are available for dogs with specific nutrient requirements caused by the presence of disease (Kirk and Bonagura, 1992; Lewis et al., 1987~. Such diets should be fed only under the supervision of . . a veterinarian. Feeding Most commercial rations are formulated to meet all nutrient require- ments if a dog eats enough to fulfill its caloric requirements. Estimates of daily caloric requirements can be obtained from several sources, including the manufacturer of the specific food being used. These estimates can be used to initiate feeding programs, but they might need substantial modifica- tion because of variations in metabolic rates of individual dogs. Under most kennel conditions, meal feeding is preferable to free-choice feeding, and individual feeding is preferable to group feeding for the fol- lowing reasons: · Restricted feeding has been shown to decrease the incidence of metabolic bone disease in growing dogs that mature at greater than 30 lb (Kealy et al., 1992). . Restricted feeding has been shown to decrease the incidence of obe- sity in young beagles and Labrador retrievers (Kendall and Burger, 1980~. · The continual ingestion of small amounts of food observed in free- choice feeding programs stimulates oral bacterial growth and might pro- mote dental disease and gingivitis (Dr. John Saidla, Department of Clinical Sciences, New York State College of Veterinary Medicine, Cornell Univer- sity, Ithaca, N.Y., unpublished). · When dogs are fed in groups, dominant dogs might overeat and might prevent subordinate dogs from eating enough to fulfill their daily needs. · When dogs are fed individually, their food intake can be monitored. Some kennels have successfully used free-choice feeding to maintain dogs. This practice is most successful when the diet used is a food of relatively low energy density and palatability. Dogs must be fed at least once a day, except as required for adequate . .. . . . . ~. . veterinary care (9 CFR 3.9a). Each healthy adult dog should be fed enough to maintain its optimal body weight; this amount will vary with the environ- ment and with the dog's age, sex, breed, temperament, and activity. Within an individual breed, there is often a wide variety of normal sizes. It is better to evaluate a dog's size according to how it looks and how it feels than according to body weight alone. With the hands-on approach, a dog's .
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26 DOGS: LABORATORY ANIMAL MANAGEMENT rib cage, spinous processes, and ileal wings should be easily palpable. They should not protrude from under the skin, nor should they be buried under a layer of adipose tissue. Once an adult dog is being maintained at its ideal body size, its weight can be used as a reference for future evaluation of food requirement. However, the loss of muscle mass and gain of adipose tissue, such as are observed in several endocrine disorders, and shifts in - fluid balance might make body weight an inaccurate means of assessing nutritional status; therefore, body weight should not completely replace ap- pearance and feel as assessment methods. _ Contaminants Animal-colony managers should be judicious in purchasing, transport- ing, storing, and handling food to ensure that it does not introduce diseases, potential disease vectors, or parasites. Food must be stored in a manner that prevents spoilage, contamination, and vermin infestation. Open bags must be stored in leakproof containers with tightly fitting lids (9 CFR 3.1e; NRC, 1985a). Contaminants in food can have dramatic effects on biochemical and physiologic processes. In general, food for dogs should not be manufac- tured or stored in facilities used for farm foods or any products containing additives, such as rodenticides, insecticides, hormones, antibiotics, fumi- gants, or other potential toxicants. WATER Ordinarily, all dogs should receive fresh, clean, potable water ad libi- tum. If water is not continuously available, the AWRs require that it be made available at least twice a day for at least 1 hour each time, unless it is restricted by the attending veterinarian (9 CFR 3.101. Watering devices can be either portable or self-watering. Self-watering devices are convenient and reduce labors but they require scheduled obser- vations to ensure proper function. Portable watering devices should be easily removable for daily rinsing and periodic sanitizing. BEDDING AND RESTING APPARATUSES Bedding can be used in some husbandry situations. For example, if drains are not available, it can be used as an absorbent to help to keep dogs clean and dry. Kinds of bedding typically used for dogs are wood shavings and shredded paper. Bedding must be stored in a manner that protects it from contamination and vermin infestation (9 CFR 3.1e). Resting apparatuses, especially those made of high-density polyethyl
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HUSBANDRY 27 ene (Britz, 1990), are useful for minimizing loss of body heat from dogs in postoperative recovery, dogs in ill health, and young pups with poorly de- veloped heat-control mechanisms. SANITATION The schedule for cleaning and disinfecting dog facilities will vary ac- cording to the physical makeup of pens, cages, or runs and other factors. Generally, primary enclosures should be cleaned as needed and sanitized at least once every 2 weeks. Excrement pans and runs should be cleaned daily. If pens and runs composed of materials that cannot be sanitized (e.g., gravel, sand, or pea stone) are used, the contaminated materials should be replaced as often as necessary to prevent odors, diseases, and vermin infes- tation. Procedures outlined in the AWRs (9 CFR 3.11) should be followed. Dogs must be removed before the floors of primary enclosures are thor- oughly cleaned. Primary enclosures containing bitches near parturition, dams with litters, or dogs in quarantine require a cleaning schedule that disturbs them as little as possible. Equipment and peripheral areas should be cleaned according to the recommendations of the Guide (NRC, 1985a). Waste should be removed regularly and frequently, and safe, sanitary procedures should be used to collect and discard it (NRC, 1985a). IDENTIFICATION AND RECORDS Identification Each dog held in a research facility must be marked either with the official USDA tag or tattoo that was on the dog at the time it was acquired or with a tag, tattoo, or collar applied by the facility that individually iden- tifies the dog by number (9 CFR 2.38gl). Unweaned puppies need not be individually numbered as long as they are maintained in the same primary enclosure as their dam (9 CFR 2.38g3~. However, they can be marked for identification with a variety of methods. Colored yarns or spots made with such marking substances as nail polish or paint provide a quick visual reference. Subcutaneous dots can be made by injecting a small amount of tattoo ink beneath the abdominal skin with a tuberculin syringe and 25-gauge needle. Ink dots should be placed in a different location for each pup (e.g., left axilla and right side of abdomen). The location or pattern of the dots and the sex and markings of each pup provide individual identification until permanent tattoos can be applied. Tattooing of the inner surface of a dog's ear is common. Before the tattoo is applied, the ear should be cleaned thoroughly. Tattoos can be
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28 DOGS: LABORATORY ANIMAL MANAGEMENT applied with special pliers or an electrovibrator. A tattoo might have to be reapplied after several years. An ancillary method for individually identify- ing dogs uses a subcutaneously implanted, permanently encoded microchip (transponder) that, when activated by an electronic scanner, broadcasts the encoded number; the scanner transfers the broadcast to a processor that produces either a digital readout or a printed copy. This identification system can be useful during daily examination of dogs being used in stud- ies, but it has not been approved by USDA as the sole source of identifica- tion because there is no standard implantation site, no standardized scanner, and no definitive information on whether the microchip migrates from the implantation site. USDA has approved the trial use of the microchips for a few commercial organizations (Richard L. Crawford, Assistant Deputy Ad- ministrator for Animal Care, Regulatory Enforcement and Animal Care, APHIS, USDA, Beltsville, Md., personal communication, 1993~. Record-Keeping Record-Keeping for Scientists and Animal-Care Staff A life-long, day-to-day log of individual events and experimental pro- cedures experienced by each dog especially surgery, postsurgical analge- sia, and other veterinary interventions-should be carefully maintained. The log will assist animal-care personnel in providing appropriate care, investi- gators in interpreting research results, and the institution in preparing its annual report to USDA (9 CFR 2.36~. Computer programs for maintaining such logs are commercially available (Riley and Blackford, 19914. For small colonies, hand-kept records on each dog might be more appropriate. McKelvie and Shultz (1964) described a record system for long-term stud- ies that is still relevant; it covers clinical examination and includes a coded daily log entry of all events that the animal has experienced. Records Required by Federal Regulations Research facilities are obliged to maintain records on procurement, transport, and disposal of all dogs and an inventory of dogs in the facility. When dogs are procured, facilities are required to obtain detailed information on the seller including name, address, USDA license or registration number or vehicle license number and state and a description of each dog (9 CFR 2.35b). Likewise, when a dog is transferred to another owner, records must include the name and address of the purchaser, the date and method of transport, and a certificate of health (9 CFR 2.35c). Additional information is available in the section of this chapter entitled "Transportation." A variety of forms are available to assist institutions in keeping records.
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HUSBANDRY 29 Among them are USDA Interstate and International Certificate of Health Examination for Small Animals (VS Form 18-1), Record of Dogs and Cats on Hand (VS Form 18-5), and Record of Disposition of Dogs and Cats (VS Form 18-6~. These forms can be obtained from Regulatory Enforcement and Animal Care, APHIS, USDA, Federal Building, Room 565, 6505 Belcrest Road, Hyattsville, MD 20782 (telephone: 301-436-7833~. All records should be maintained for at least 3 years (9 CFR 2.35f). Records must also be maintained on all offspring born to dogs in the colony (9 CFR 2.35b) and on exceptions to the requirements for exercise (9 CFR 3.8d). Facilities conducting research on any vertebrate animal, includ- ing dogs, are obliged to maintain additional records that include the follow- ~ng: · minutes of meetings of the IACUC; · semiannual IACUC reports; · protocols involving animal use; · scientifically justified deviations from the AWRs; and · studies involving pain in which analgesics cannot be used. Some of the information must be reported annually to USDA (9 CFR 2.361; other information, such as approved protocols, must be maintained for 3 years after the study ends (9 CFR 2.35f). EMERGENCY, WEEKEND, AND HOLIDAY CARE Dogs should be observed and cared for by qualified personnel every day, including weekends and holidays, as outlined in the Guide (NRC, 1985a). Emergency veterinary care should be available after working hours and on weekends and holidays. For dogs undergoing particular experimental pro- cedures and dogs with conditions that might require emergency care, inves- tigators should develop written protocols and provide appropriate additional coverage. TRANSPORTATION Transportation over long distances is known to be a stressor for ani- mals. Proper attention to environmental conditions, cage design, and care in transit will minimize the stress. The AWRs specify the requirements for transporting dogs (9 CFR 3.13-3.19~. Before a dog is transported, special arrangements must be made between the shipper (consignor), the carriers) or intermediate handlers, and the recipient (consignee). The shipper must certify that the dog was offered food and water during the 4 hours before delivery to the carrier and must prepare a written certification, which must
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30 DOGS: LABORATORY ANIMAL MANAGEMENT be securely attached to the cage and must contain the shipper's name and address, the animal identification number, the time and date when the dog was last offered food and water, specific instructions for feeding and water- ing the dog for a 24-hour period, and the signature of the shipper with the date and time when the certification was signed. Primary Enclosures Carriers must not accept dogs for shipment if their primary enclosures do not meet the requirements of the AWRs (9 CFR 3.14~. The primary enclosure must be large enough to allow a dog to turn around while stand- ing, to stand and sit erect, and to lie in a natural position. Primary enclo- sures must be structurally sound, free of internal protrusions that could cause injury, constructed of nontoxic materials, and able to withstand the normal rigors of transportation. The container must secure the animal and all parts of its body inside the enclosure. Devices, such as handles, must be attached to the outside to allow the container to be lifted without tilting. The container must have a leakproof, solid floor or have a raised floor and a leakproof collection tray. If animals are housed directly on the floor, absor- bent bedding material must be provided. Primary enclosures must be cleaned and any litter replaced if dogs are in transit for more than 24 hours. Pri- mary enclosures should be well ventilated to minimize the potential for a thermal gradient during shipment. Additional specifications for transport cages are in the AWRs (9 CFR 3.14) and the IATA Live Animal Regulations (IATA, 1993 et send. Puppies 4 months old or younger must not be transported in the same primary enclosure with adult dogs other than their dams. For puppies shipped during sensitive periods of behavioral development (i.e., 8-14 weeks of age; see Scott and Fuller, 1965), shipping stress should be minimized. Dogs likely to display aggressive behavior must be shipped individually, and females in heat must not be transported in the same primary enclosures as males. No more than two live puppies 8 weeks to 6 months old, of compa- rable size, and weighing 9 kg (20 lb) or less each may be transported by air in the same primary enclosure. Older dogs and puppies weighing more than 9 kg (20 lb) should be individually housed. Weaned littermates that are less than 8 weeks old and are accompanied by their dam may be transported in the same enclosure to research facilities, either by air or surface transport. During transport by surface vehicle, no more than four dogs 8 weeks old or older and of comparable size may be transported in the same primary enclo sure. When viral-antibody-free (unvaccinated) dogs are transported between facilities, precautions must be taken to avoid contact with infectious agents. Some commercial suppliers have developed filtered shipping containers to
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HUSBANDRY 31 transport those dogs. IATA rules require that special measures be taken to ensure that ventilation rates are maintained within the container, that the container be appropriately labeled, that sufficient water be provided for the entire journey, and that food, if required, be provided at the point of origin (IATA, 1993~. Environmental Conditions At all times, containers holding dogs should be placed in climate-con- trolled areas that provide protection from the elements (9 CFR 3.13, 3.15, 3.18-3.19~. Trucks and planes must be ventilated and provide air that has adequate oxygen and is free of harmful gases and particulate contaminants. Airlines should always place dogs in pressurized compartments. Dogs may be shipped if temperatures will fall below 7.2°C (45°F) during any portion of their journey only if a veterinarian certifies in writing that they have been acclimated to lower temperatures and states the lowest temperature to which they have been acclimated. During transit, dogs must not be exposed to ambient temperatures exceeding 29.4°C (85°F) for a period of more than 4 hours. Food and Water All dogs must be offered food and water within 4 hours of delivery to the carrier (9 CFR 3.13c). Carriers must offer water to each dog at 12-hour intervals beginning 12 hours after the shipper last offered water. Adult dogs must be fed at least once every 24 hours, and puppies less than 16 weeks old must be fed every 12 hours throughout the trip. Feeding and watering utensils must be firmly secured to the inside of the container and placed so that they can be filled from outside the container. Written in- structions for feeding and watering in transit must be attached to the pri- mary enclosure in such a way that they are easily seen and read (9 CFR 3.16). Other Requirements There are special requirements for animal holding areas of terminal facilities, including rules for sanitization, pest control, ventilation, tempera- ture control, and shelter from direct sunlight, rain, snow, and extreme heat (9CFR3.18). Each dog must be accompanied by a health certificate, issued by a licensed veterinarian not more than 10 days before shipping, that states that the dog is free of any infectious disease or physical abnormality that would endanger it or other animals or pose a threat to public health. An exemp
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32 DOGS: LABORATORY ANIMAL MANAGEMENT lion can be made by the secretary of USDA for individual animals shipped to research facilities if the facilities require animals that are not elegible for certification (9 CFR 2.78b). Instructions for the administration of drugs or provision of other special care must be firmly attached to the outside of the container (9 CFR 3.14h). A pregnant bitch should be accompanied by a certificate, signed by a veterinarian, that states that there is no risk of birth during transit (IATA, 1993~. Carriers and intermediate handlers must not accept dogs more than 4 hours before the scheduled departure (6 hours by special arrangement). An attempt must be made to notify the recipient on arrival at the destination and at least once every 6 hours thereafter (9 CFR 3.13f). During shipment by surface transportation, the operator of the conveyance or someone ac- companying the operator must observe the dogs at least once every 4 hours to ascertain that they have sufficient air for normal breathing and are not in distress and that the rules for ambient temperature and all other AWR re- quirements are met. The same rules apply in air carriers if the animal cargo area is accessible during flight. If it is not accessible, the carrier must observe the dogs at loading and unloading. Dogs in physical distress must receive veterinary care as soon as possible (9 CFR 3.17~. REFERENCES AAFCO (Association of American Feed Control Officials), Canine Nutrition Expert Subcom- mittee, Pet Food Committee. 1993. AAFCO nutrient profiles for dog foods. Pp. 92-99 in Official Publication 1993. Atlanta: Association of American Feed Control Officials. Available from Charles P. Frank; AAFCO Treasurer; c/o Georgia Department of Agricul- ture; Plant Food, Feed, and Grain Division; Capitol Square, Atlanta, GA 30334. Bellhorn, R. W. 1980. Lighting in the animal environment. Lab. Anim. Sci. 30(2): 440-450. Besch, E. L. 1985. Definition of laboratory animal environmental conditions. Pp. 297-315 in Animal Stress, G. P. Moberg, ed. Bethesda, Md.: American Physiological Society. Besch, E. L. 1990. Environmental variables and animal needs. Pp. 113-131 in The Experi- mental Animal in Biomedical Research. Vol. I: A Survey of Scientific and Ethical Issues for Investigators, B. E. Rollin and M. L. Kesel, eds. Boca Raton, Fla.: CRC Press. Besch, E. L. 1991. Temperature and humidity control. Pp. 154-166 in Handbook of Facilities Planning. Vol. 2: Laboratory Animal Facilities, T. Ruys, ed. New York: Von Nostrand Reinhold. Besch, E. L. 1992. Animal facility ventilation air quality and quantity. ASHRAE Trans. 98(pt. 2):239-246. Britz, W. E., Jr. 1990. Caging systems for dogs under the new standards of the Animal Welfare Act. Pp. 48-50 in Canine Research Environment, J. A. Mench and L. Krulisch, eds. Bethesda, Md.: Scientists Center for Animal Welfare. Available from SCAW, 4805 St. Elmo Avenue, Bethesda, MD 20814. Campbell, S. A., H. C. Hughes, H. E. Griffin, M. S. Landi, and F. M. Mallon. 1988. Some effects of limited exercise on purpose-bred beagles. Am. J. Vet. Res. 49:1,298-1,301. Clark, J. D., J. P. Calpin, and R. B. Armstrong. 1991. Influence of type of enclosure on exercise fitness of dogs. Am. J. Vet. Res. 52:1,024-1,028. Clough, G., and M. R. Gamble. 1976. Laboratory Animal Houses. A Guide to the Design and
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HUSBANDRY 33 Planning of Animal Facilities. LAC Manual Series No. 4. Carshalton, Surrey, U.K.: Medical Research Council Laboratory Animals Centre. 44 pp. Edwards, R. G., M. F. Beeson, and J. M. Dewdney. 1983. Laboratory animal allergy: The measurement of airborne urinary allergens and the effect of different environmental con- ditions. Lab. Anim. (London) 17:235-239. Fuller, J. L. 1970. Genetic influences on socialization. Pp. 7-18 in Early Experiences and the Process of Socialization, R. A. Hoppe, G. A. Milton, and E. C. Simmel, eds. New York: Academic Press. Gorton, R. L., and E. L. Besch. 1974. Air temperature and humidity response to cleaning water loads in laboratory animal storage facilities. ASHRAE Trans. 80(pt. 1):37-52. Gorton, R. L., J. E. Woods, and E. L. Besch. 1976. System load characteristics and estimation of annual heat loads for laboratory animal facilities. ASHRAE Trans. 82(pt. 1):107-112. Hetts, S., J. D. Clark, J. P. Calpin, C. E. Arnold, and J. M. Mateo. 1992. Influence of housing conditions on beagle behaviour. Appl. Anim. Behav. Sci. 34:137-155. Hite, M., H. M. Hanson, N. R. Bohider, P. A. Conti, and P. A. Mattis. 1977. Effect of cage size on patterns of activity and health of beagle dogs. Lab. Anim. Sci. 27:60-64. Hubrecht, R. C. 1993. A comparison of social and environmental enrichment methods for laboratory housed dogs. Appl. Anim. Behav. Sci. 37:345-361. Hubrecht, R. C., J. A. Serpell, and T. B. Poole. 1992. Correlates of pen size and housing conditions on the behaviour of kennelled dogs. Appl. Anim. Behav. Sci. 34:365-383. Hughes, H. C., and S. Campbell. 1990. Effects of primary enclosure size and human contact. Pp. 66-73 in Canine Research Environment, J. A. Mench and L. Krulisch, eds. Bethesda, Md.: Scientists Center for Animal Welfare. Available from SCAW, 4805 St. Elmo Avenue, Bethesda, MD 20814. Hughes, H. C., S. Campbell, and C. Kenney. 1989. The effects of cage size and pair housing on exercise of beagle dogs. Lab. Anim. Sci. 39:302-305. IATA (International Air Transport Association). 1993. IATA Live Animal Regulations, 20th ed. Montreal, Quebec: International Air Transport Association. Available from IATA, Publications Department, 2000 Peel Street, Montreal, Quebec, Canada H3A 2R4. Kealy, R. D., S. E. Olsson, K. L. Monti, D. F. Lawler, D. N. Biery, R. W. Helms, G. Lust, and G. K. Smith. 1992. Effects of limited food consumption on the incidence of hip dyspla- sia in growing dogs. J. Am. Vet. Med. Assoc. 201:857-863. Kendall, P. T., and I. H. Burger. 1980. The effect of controlled and appetite feeding on growth and development in dogs. Pp. 60-63 in Proceedings of the Kal Kan Symposium for the Treatment of Dog and Cat Diseases (Sept. 29-30, 1979), R. L. Wyatt, ed. Vernon, Calif.: Kal Kan Foods, Inc. Available from Kal Kan Foods, Inc., 3250 E 44th Street, Vernon, CA 90058-0853. King, J. A. 1954. Closed social groups among domestic dogs. Proc. Am. Philos. Soc. 98:327- 336. Kirk, R. W., and J. D. Bonagura, eds. 1992. Current Veterinary Therapy. XI. Small Animal Practice. Philadelphia: W. B. Saunders. 1,346 pp. Lewis, L. D., M. L. Morris, Jr., and M. S. Hand. 1987. Small Animal Clinical Nutrition III. Topeka, Kans.: Mark Morris Associates. Available from Mark Morris Associates, 5500 SW 7th Street, Topeka, KS 66606. McKelvie, D. H., and F. T. Shultz. 1964. Methods of observing and recording data in long- term studies on beagles. Lab. Anim. Care 14:118-124. Neamand, J., W. T. Sweeney, A. A. Creamer, and P. A. Conti. 1975. Cage activity in the laboratory beagle: A preliminary study to evaluate a method of comparing cage size to physical activity. Lab. Anim. Sci. 25: 180- 183. Newberne, P. M., and J. G. Fox. 1978. Chemicals and toxins in the animal facility. Pp. 118- 141 in Laboratory Animal Housing. Proceedings of a symposium organized by the Insti
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34 DOGS: LABORATORY ANIMAL MANAGEMENT lute of Laboratory Animal Resources Committee on Laboratory Animal Housing. Wash- ington, D.C.: National Academy of Sciences. Newton, W. M. 1972. An evaluation of the effects of various degrees of long-term confine- ment on adult beagle dogs. Lab. Anim. Sci. 22:860-864. NRC (National Research Council), Institute of Laboratory Animal Resources, Committee on Care and Use of Laboratory Animals. 1985a. Guide for the Care and Use of Laboratory Animals. NIH Pub. No. 86-23. Washington, D.C.: U.S. Department of Health and Human Services. 83 pp. NRC (National Research Council), Board on Agriculture, Subcommittee on Dog Nutrition, Committee on Animal Nutrition. 1985b. Nutrient Requirements of Dogs, revised ed. Washington, D.C.: National Academy Press. 79 pp. Pekrul, D. 1991. Noise control. Pp. 166-173 in Handbook of Facilities Planning. Vol. 2: Laboratory Animal Facilities, T. Ruys, ed. New York: Von Nostrand Reinhold. Peterson, E. A. 1980. Noise and laboratory animals. Lab. Anim. Sci. 30:422-439. Poffenbarger, E. M., M. L. Chandler, S. L. Ralston, and P. N. Olson. 1990. Canine neonatol- ogy. Part 1. Physiologic differences between puppies and adults. Compend. Cont. Educ. Pract. Vet. 12: 1601-1609. Riley, R. D., and R. K. Blackford. 1991. ALACARTE-An animal in-life tracking system. AALAS Bull. 30(3):20-23. Available from the American Association for Laboratory Animal Science, 70 Timber Creek Drive, Suite 5, Cordova, TN 38018. Ruckenbusch, Y. 1975. The hypnogram as an index of adaptation of farm animals to changes in their environment. Appl. Anim. Ethol. 2:3-18. Scott, J. P., and J. L. Fuller. 1965. Genetics and the Social Behavior of the Dog. Chicago: University of Chicago Press. 468 pp. Sierens, S. E. 1976. The Design, Construction, and Calibration of an Acoustical Reverbera- tion Chamber for Measuring the Sound Power Levels of Laboratory Animals (thesis for M.S. degree). Gainesville: University of Florida. 127 pp. Available from Health Science Center Library, University of Florida, Box 100206, Gainesville, FL 32610-0206. Stanley, W. C. 1965. The passive person as a reinforcer in isolated beagle puppies. Psychon. Sci. 2:21-22. Stanley, W. C., and O. Elliot. 1962. Differential human handling as reinforcing events and as treatment influencing later social behavior in basenji puppies. Psychol. Rep. 10:775-788. Wolfle, T. L. 1990. Policy, program and people: The three P's to well-being. Pp. 41-47 in Canine Research Environment, J. A. Mench and L. Krulisch, eds. Bethesda, Md.: Scien- tists Center for Animal Welfare. Available from SCAW, 4805 St. Elmo Avenue, Bethesda, MD 20814.
Representative terms from entire chapter: