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Laboratory Animal Management: Dogs (1994)

Chapter: 6 SPECIAL CONSIDERATIONS

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Suggested Citation:"6 SPECIAL CONSIDERATIONS." National Research Council. 1994. Laboratory Animal Management: Dogs. Washington, DC: The National Academies Press. doi: 10.17226/2120.
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6 Special Considerations PROTOCOL REVIEW One of the many important responsibilities of an institutional animal care and use committee (IACUC) is to review the protocols of research projects in which dogs will be used (9 CFR 2.31; PHS, 1986~. The proto- col-review mechanism is designed to ensure that investigators consider the care and use of their animals and that protocols comply with federal, state, and institutional regulations and policies. In addition, the review mecha- nism enables an IACUC to become an important institutional resource, as- sisting investigators in all matters involving the use of animals. Although the discussion below is directed to the use of dogs in research, the review requirements apply to all vertebrate species. Each research protocol must completely (but concisely) delineate the proposed study, including a description of each of the following: · the purpose of the study; · the rationale for selecting dogs as the research subjects; · the breed, age, and sex of the dogs to be used; · the numbers of dogs in various groups of the protocol and the total number to be used; · experimental methods and manipulations; · experimental manipulations that will be performed repeatedly on an individual dog; 76

SPECIAL CONSIDERATIONS · preprocedural and postprocedural care and medications; · procedures that will be used to minimize discomfort, pain, and dis- tress, including, where appropriate, the use of anesthetics, analgesics, tran- quilizers, and comfortable restraining devices; · the euthanasia method, including the reasons why it was selected and whether it is consistent with the recommendations of the American Veteri- nary Medical Association Panel on Euthanasia (AVMA, 1993, et seq.~; · the process undertaken to ensure that there are no appropriate in vitro alternatives, that there are no alternative methods that would decrease the number of animals to be used, and that the protocol does not unneces- sarily duplicate previous work; and · the qualifications of the investigators who will perform the proce- dures outlined. One approach used by IACUCs is to have a scientifically knowledge- able member thoroughly review the protocol. The reviewer contacts the investigator directly to clarify issues in question. Later, at an IACUC meet- ing, the reviewer presents and discusses the protocol and relates discussions with the investigator. Changes or clarifications in the protocol that have resulted from the reviewer's discussions with the investigator are submitted to the IACUC in writing. After presentation of the protocol, the reviewer recommends a course of action, which is then voted on by the IACUC. Another kind of protocol review (which is especially effective in small institutions with few grants) is initial review by the entire IACUC; results are generally available to the investigator within a short period. Several outcomes of protocol review are possible: approval, approval contingent on receipt of additional information (to respond to minor prob- lems with the protocol), deferral and rereview after receipt of additional information (to respond to major problems with the protocol), and with- holding of approval. If approval of a protocol is withheld, an investigator should be accorded due process and be given the opportunity to rebut the IACUC's critique in writing, to appear in person at an IACUC meeting to present his or her viewpoint, or both. It is also important that provision be made for expedited review, in which a decision is reached within 24-48 hours. Expedited reviews should be used only for emergency or extenuat- ing circumstances. When a protocol is submitted for expedited review, each member of the IACUC must have an opportunity to review it and may call for a full committee review before approval is given and before animal work begins (McCarthy and Miller, 1990~. The question of protocol review for scientific merit has been handled in a variety of ways by IACUCs. Many protocols are subjected to extensive, external scientific review as part of the funding process; in such instances, the IACUC can be relatively assured of appropriate scientific review. In 77

78 DOGS: LABORATORY ANIMAL MANAGEMENT the case of studies that will not undergo outside review for scientific merit, many IACUCs require signoff by the investigators, department chairmen, or internal review committees; this makes the signer responsible for providing assurance that the proposed studies have been designed and will be per- formed "with due consideration of their relevance to human or animal health, the advancement of knowledge, or the good of society" (NRC, 1985, p. 82; PHS, 1986, p. 27~. Occasionally, IACUC members and investigators differ as to the relevance of proposed studies to human and animal health and the advancement of knowledge. Each institution should develop guidelines for dealing with this potential conflict. RESTRAINT Some form of restraint is generally necessary to control a dog during a procedure (see guidelines in NRC, 1985, p. 9~. The method used should provide the least restraint required to allow the specific procedure to be performed properly, should protect both the dogs and personnel from harm, and should avoid causing distress, physical harm, or unnecessary discom- fort. In handling and restraining dogs, it is helpful to understand species- typical behavior patterns and communication systems. A small or medium-size dog can be picked up by placing one hand under the chest and abdomen while restraining the head with a leash. Lift- ing a large dog might require two people. It is important to remember that males are sensitive to touch near their genitalia. Minor procedures, such as taking a rectal temperature or administering a subcutaneous injection, can usually be accomplished by one person using minimal restraint. During venipuncture, sufficient restraint should be used to avoid repeated needle insertions and to prevent the development of painful hematomas. Kesel and Neil (1990) detail methods for handling and restraining animals. If dogs are to be restrained frequently or for long periods or if the restraint method used is especially rigorous, it might be necessary to train them to tolerate the restraint. Training sessions should use positive-rein- forcement techniques; negative-reinforcement techniques are not desirable. Physical abuse (9 CFR 2.38f2i) and food or water deprivation (9 CFR 2.38f2ii) must not be used to train, work, or handle dogs, although food and water may be withheld for short periods when specified in an IACUC-approved protocol (9 CFR 2.38f2ii). SPECIAL CARE FOR ANIMAL MODELS The remainder of this chapter deals with some common uses of labora- tory dogs in which aspects of care vary from the general guidelines pro- vided in previous chapters. It is not intended to present an exhaustive list

SPECIAL CONSIDERATIONS 79 of canine models that require special housing and husbandry, but rather to provide the reader with different types of canine models that can serve as examples of how housing and husbandry can be modified to achieve animal well-being. The suggestions offered here are not to be construed as the only ones possible. The committee recognizes that not every research pro- cedure and circumstance can be anticipated, and it assumes that sound pro- fessional judgment, good veterinary practices, and adherence to the spirit of this guide will prevail in unusual situations. The final subsection of this chapter introduces the reader to the tech- nique of somatic cell gene therapy. Many disorders of dogs, like those of humans, are caused by single-gene mutations. Scientists are working to develop techniques to cure these disorders permanently by replacing mutant genes with normal ones. For many reasons (see Chapter 2), the dog is an ideal model for evaluating the safety and efficacy of gene therapy. Aging Clinical Features Life expectancy and disease incidences vary among breeds of dogs; therefore, it is not possible to state a specific age at which dogs become old. Common laboratory dogs, such as beagles, begin some aging changes when they are 8-10 years old. Such physical features as graying of the haircoat, especially around the face, are often apparent as aging begins. As dogs age, they tend to become less active and to exhibit such signs of mental deterioration as poor recognition of caretakers, excessive sleep- ing, and changes in personality. Senile plaques, similar to those found in humans with senile dementias, have been reported in the brains of old dogs (Wisniewski et al., 1970~. Various forms of arthritis, spondylosis, and degenerative joint disease are common and contribute to problems in mobil- ity and to the apparent diminution of mental alertness. Older dogs might decrease their daily food intake, become slow eaters, or become irregular in their eating habits. Dental problems including periodontal disease, tooth abscesses, and oral-nasal fistulas increase; the importance of these prob- lems is probably underestimated (Tholen and Hoyt, 1983~. Dogs more than 6 years old develop lenticular sclerosis, which results in a bluish appear- ance within the pupil. Visual acuity decreases with age and is often associ- ated with cataracts, secondary glaucoma, and other diseases (Fischer, 1989~. There is also apparent hearing loss. Atrophy of the thyroid gland and an increased number of thyroid tu- mors have been reported, and signs of hypothyroidism are common (Haley et al., 1989; Milne and Hayes, 19811. Thyroid atrophy and the propensity of older dogs to develop hypothermia might be related (B. A. Muggenburg, l

80 DOGS: LABORATORY ANIMAL MANAGEMENT Inhalation Toxicology Research Institute, Lovelace Biomedical and Envi- ronmental Research Institute, Albuquerque, N.M., unpublished). A decreased response to antigens and changes in lymphocyte function might indicate that the older dog is less able to resist infectious diseases (Bice and Muggenburg, 1985~. Some changes in common blood-cell measures and serum chemistry become important when these are used for diagnosis (Lowseth et al., 1990a). The incidence of neoplasia increases strikingly (MacVean et al., 1978~; for example, lung tumors, nearly unknown in young dogs, can reach an inci- dence as high as 10 percent in dogs over 10 years old (Ogilive et al., 1989~. Pulmonary function decreases with age because of reduced lung volumes and decreased elasticity (Mauderly and Hahn, 1982~. Chronic renal dis- eases often occur and require frequent monitoring. Chronic heart disease is also fairly common, and clinical signs can appear suddenly in old dogs. Husbandry and Veterinary Care Housing and environment. Accommodation should be made for dogs that have problems moving comfortably on floor grates or through guillo- tine-like doors in kennel buildings. Because of their decreased mobility and impaired thermoregulatory function, aging dogs with access to outdoor ar- eas should be checked frequently to be certain that they are able to get inside to escape the cold or heat. Automatic watering devices might be- come difficult to use; for some old dogs, it might be necessary to switch to water pans placed on the floor. Nutrition. Differentiation between age-related and disease-caused changes in eating habits might be difficult. It is important that animal-care person- nel become familiar with and closely monitor daily eating habits of older dogs. Frequent checking and recording of body weights can help in assess- ing whether food intake is adequate. Changes in diet are sometimes dic- tated by the clinical diagnosis of disease (e.g., a low-protein diet for chronic, progressive renal disease and a low-sodium diet for chronic heart failure). Physical characteristics of food can affect dental hygiene. Soft and wet food fed over many years can contribute to dental disease. Feeding dry dog food and providing hard objects for chewing can be helpful in the long-term management of dental problems. Routine dental care, including the removal of calculus and polishing, is essential. Veterinary care. The extent of chronic disease problems in older dogs requires more intensive veterinary care, extensive diagnostic investigations, and good nursing. Dosages of some medications might have to be reduced, because drugs are commonly metabolized more slowly in old than in young adult dogs. Such drugs as digoxin should be monitored by measuring blood

SPECIAL CONSIDERATIONS 81 concentrations to decrease the risk of overdosing (De Rick et al., 1978~. A useful reference on geriatric veterinary medicine is Geriatrics and Geron- tology (Goldston, 19899. Reproduction Bitches. Andersen and Simpson (1973) have described reproductive senescence in beagle bitches. Intact bitches exhibit irregular estrous cycles, accompanied by decreased fertility, and prolonged periods of anestrus. The mortality rate is higher among puppies born to older bitches than among puppies born to bitches less than 3 years old. The most common pathologic condition of the uterus of aged bitches is pyometra (Andersen and Simpson, 1973; Jarvinen, 1981; Whitney, 1967~. Vaginal fibromuscular polyps are also common (Andersen and Simpson, 1973~. The age-specific incidence of mammary gland neoplasms in intact beagle bitches continues to increase throughout life (Taylor et al., 1976~. Dogs. Aging dogs have testicular atrophy and often develop prostatic hypertrophy and hyperplasia and have episodes of prostatitis (Lowseth et al., l990b). There are also metaplastic changes in the bladder (Lage et al., 1989~. Cardiovascular Diseases Congenital Heart Defects Clinical Features Dogs with hereditary cardiovascular malformations have been used to investigate the role of genetic and embryologic factors in the cause and pathogenesis of congenital heart defects, including hereditary patent ductus arteriosus, conotruncal defects (e.g., ventricular septal defect, tetralogy of Fallot, and persistent truncus arteriosus), discrete subaortic stenosis, and pulmonary valve dysplasia. Congenital heart defects in dogs have been summarized by Buchanan (1992) and Eyster (19923. Table 6.1 describes and lists the clinical signs of selected heart defects. Each of those defects is transmitted as a lesion-specific genetic defect in one or more breeds. A model for each defect has been developed at the University of Pennsylvania School of Veterinary Medicine by selective breeding of affected dogs (Patterson, 1968), as follows: patent ductus arteriosus, toy and miniature poodles (Acker- man et al., 1978; De Reeder et al., 1988; Gittenberger-de Groot et al., 1985; Knight et al., 1973; Patterson et al., 1971~; conotruncal defects, keeshonden (Patterson et al., 1974, 1993; Van Mierop et al., 1977~; discrete subaortic

82 DOGS: LABORATORY ANIMAL MANAGEMENT TABLE 6.1 Selected Congenital Cardiac Defects in Dogs Defect Description Clinical Signs Patent ductus Failure of ductus arteriosus to Vary with size of duct and arteriosus close after birth. If pulmo- pulmonary vascular resistance nary vascular resistance is from subclinical to heart low, blood flows through failure. Early signs include ductus from left to right. Pul- poor growth, coughing, and monary hypertension and left dyspnea. Aneurysm can occur ventricular hypertrophy result at site of ductus arteriosus. unless ductus opening is small. Polycythemia occurs in If ductus is large and pulmonary cyanotic dogs with a large vascular resistance is high, put- patent ductus arteriosus lmonary arterial pressure can (PDA), pulmonary hyper exceed aortic pressure, and blood tension, and right to left will flow from right to left, sending blood flow through the PDA. venous blood into ascending aorta. Conotruncal defects Ventricular Failure to complete formation of Vary with size of defect from septal the conotruncal septum results in subclinical to signs of defect ventricular septal defects (VSDs) respiratory and right-side of varied size, involving the lower heart failure, including and middle portions of the crista cyanosis, dyspnea, weakness, supraventricularis (Type I, sub- and anorexia. arterial VSD). Pups with large VSDs usually die from pulmonary edema in the neonatal period. Smaller VSDs are compatible with long life unless complicated by pulmonary hypertension and congestive heart failure. Tetralogy of Consists of pulmonic stenosis Fallot (valvular, infundibular, or both), conal ventricular septal defects, dextroposition of aorta with overriding of ventricular septum, and right ventricular hypertrophy. Some dogs have pulmonary valve atresia (pseudo-truncus arteriosus). Persistent truncus arteriosus Depend on severity of pul . . . mon1c stenosis anc ventrlc- cular septal defect. Can include decreased body size, fatigue, cyanosis, and secondary polycythemia. Severe but rare anomaly. Complete Cyanosis and dyspnea. Dogs failure of septation of conus and rarely survive neonatal period. truncus regions, producing large conal ventricular septal defect and single arterial outlet vessel.

SPECIAL CONSIDERATIONS TABLE 6.1 Continued 83 Defect Description Clinical Signs Discrete subaortic Narrowing of left ventricular out- Vary with degree of stenosis stenosis flow tract, most commonly by from asymptomatic to poor fibrous ring just below aortic growth, exercise intolerance, semilunar valves, with concomi- syncope, ventricular tent obstruction of blood flow, arrhythmias, pulmonary left ventricular hypertrophy, edema, and sudden death. and increased left ventricular pressure. Pulmonary valve Varies from mild thickening of Vary from asymptomatic to dysplasia leaflets surrounding narrowed dyspnea, fatigability, and pulmonary orifice to complete right-side heart failure. fusion of leaflets and doming of valve. Interferes with emptying of right ventricle. stenosis, Newfoundlands (Patterson, 1984; Pyle et al., 1976), and pulmo- nary valve dysplasia, beagles (Patterson, 1984; Patterson et al., 1981~. Conotruncal defects in the keeshond breed are determined by the effect of a single major gene defect (Patterson et al., 19931. Subaortic stenosis in Newfoundlands also appears to be monogenic with variable expression (Patterson, 1984~. Patent ductus arteriosus and pulmonary valve dysplasia are inherited in a non-Mendelian pattern. Husbandry and Veterinary Care Animals with cardiac defects often require exercise restriction to avoid cyanosis and congestive heart failure. The need for restriction must be decided for each dog on the basis of cardiac status. If the clinical manifes- tations of severe defects (e.g., respiratory distress, severe cyanosis, and congestive heart failure) cannot be relieved with appropriate surgical meth- ods or cardiovascular drugs (e.g., cardiac glycosides and diuretics), the dog should be humanely killed (see Chapter 5~. Reproduction Only dogs with mild to moderate cardiac defects or those in which the defects have been surgically corrected should be selected for breeding. Se- verely affected dogs do not survive to breeding age, or they develop clinical manifestations that preclude their use for reproduction (e.g., marked cyanosis

84 DOGS: LABORATORY ANIMAL MANAGEMENT and congestive heart failure). Methods of modern clinical cardiology- including auscultation, radiography, echocardiography, cardiac catheterization, and angiocardiography are necessary for accurate diagnosis and evalua- tion of the severity of defects in candidates for breeding. Therefore, appro- priate facilities and equipment and personnel qualified to use such equip- ment must be available before a breeding colony is established. Once it is established, the health status of breeding stock and their offspring must be carefully monitored. Induced Heart Defects Clinical Features Many animal models of cardiac disease are surgically induced in physi- ologically normal animals. Aims of the research protocol and humane con- siderations must often be carefully balanced to ensure that the maximal amount of information is derived from each animal. Surgically induced models can be broadly divided into models of vol- ume or pressure overload produced by creating valvular or interchamber defects, models of ischemic injury, and models of arrhythmia (Gardner and Johnson, 1988~. Long-term management of these models Carl be difficult because they are frequently on the verge of physiologic decompensation and at risk of sudden death. Table 6.2 lists the signs of cardiac failure. TABLE 6.2 Clinical Signs of Heart Failure in Dogs Type of Heart Failure Clinical Signs Left-side Right-side Generalized Exercise tolerance decreases. Inappropriate dyspnea follows exercise. Pulmonary venous pressure increases, initially causing pulmonary and bronchial congestion and reflexogenic bronchoconstriction. Repetitive coughing follows exercise. Orthopnea, with a reluctance to lie down; restlessness at night; and paroxysmal dyspnea are common. In severe failure, pulmonary edema, severe dyspnea at rest, and rates on auscultation become evident. Systemic venous congestion occurs with engorgement of jugular veins. Liver and spleen are enlarged and often palpable. Fluid retention is usually first manifested as ascites; subcutantous edema, hydrothorax, or hydropericardium can follow. Disturbances of gastrointestinal function, with diarrhea, can occur. Signs of both left- and right-side failure occur.

SPECIAL CONSIDERATIONS Husbandry and Veterinary Care 85 The management of chronic dog models of induced heart failure is most successful if the approach used is interdisciplinary, involving cardiologists, surgeons, and veterinary-care staff. Goals of long-term management in- clude identifying potential complications, selecting therapeutic regimens, and developing long-term monitoring protocols. The following general guidelines should be tailored to the type of disorder induced, the dogs' well-being, and the goals of the research protocol. Postoperative care. Postoperative care depends on the type of heart disease induced. Medical management should continue after successful recovery from surgery because a specific surgical protocol does not always produce a physiologically consistent model. Some dogs achieve a stable, compensated postoperative condition; others undergoing the same proce- dure develop signs of acute heart failure immediately after surgery. Careful monitoring on the days after surgery is critical. Meticulous physical examinations should be performed on physiologically stable dogs at least once a day until they have recovered from surgery. Physiologically unstable dogs should be examined more often. Vital signs should be moni- tored, and particular attention should be given to physical findings related to the cardiovascular system. Mucous membrane color, capillary-refill time, and temperature of extremities can be abnormal if peripheral perfusion is seriously impaired. The pulse quality of the femoral artery can be used to assess systemic perfusion. Auscultation should be used to detect abnormal cardiac sounds, and electrocardiography should be performed to diagnose arrhythmias. Assessment of respiratory rate and depth should be combined with careful auscultation of all lung fields to detect early signs of pulmo- nary complications. Echocardiography, if available, can be used to evaluate cardiac function and contractility. Good nursing care is important. Special diets, such as canned dog food or dry food mixed with chicken broth, can be offered to encourage food intake. Ideally, dogs should be housed in a dedicated recovery room and returned to the regular housing area only when they are physiologically stable and have recovered fully from surgery. Decreased exercise tolerance secondary to diminished cardiac reserve might affect the extent of activity that a dog can withstand. Complications. Potential complications associated with surgical and catheterization procedures should be anticipated, including infection of the operative sites bacteremia, and endocarditis. Dogs at high risk for compli- cations are the ones that undergo serial catheterization procedures and those with bioimplants, such as prosthetic valves and pacemakers (Dougherty,

86 DOGS: LABORATORY ANIMAL MANAGEMENT 1986~. Baseline monitoring should include scheduled physical examina- tions and complete blood counts (CBCs). A blood culture should be sub- mitted to the laboratory for any animal with a persistent fever or an inter- mittently increased temperature. If infection is suspected, a broad-spectrum antibiotic, such as one of the cephalosporins, should be administered pend- ing receipt of culture and sensitivity results. Banding of the great vessels with various materials is a standard proce- dure for producing volume- and pressure-overload models of ventricular hypertrophy, coarctation of the aorta, and obstruction of right ventricular outflow. Vessel erosion caused by the material used (Gardner and Johnson, 1988) and hemorrhage secondary to banding procedures are common com- plications that should be included in the differential diagnosis of any banded animal that suffers an acute onset of lethargy, paleness of the mucous mem- branes, or dyspnea. Those are also clinical signs of heart failure, so it is important to perform auscultation of the chest and suitable diagnostic tests, such as radiography or thoracentesis, to make an accurate diagnosis. A dog that is hemorrhaging should be euthanatized. Surgical procedures used to induce cardiac disease invariably cause disruption of the endothelium and put the dogs at risk for thrombosis and embolism. Dogs undergoing cardiac catheterization or surgery of the car- diac valves are at greatest risk. Clinical signs reflect the organs involved. tong-term monitoring. In a study of extended duration, assessment of each dog's general health and cardiovascular system should be continuous. The type and frequency of examinations will depend on whether the model is physiologically stable or unstable. For example, a dog with induced mitral regurgitation, which is defined as a 50 percent reduction in forward stroke volume and a pulmonary capillary wedge pressure of 20 mm Hg, can develop life-threatening pulmonary edema (Nakano et al., 1991; Swindle et al., 1991~. Frequent monitoring and auscultation are required to detect early signs of respiratory compromise so that the dog will not die before therapy can be initiated or the dog can be studied. Similarly, a dog with induced right ventricular pressure overload requires frequent monitoring because decreased coronary blood flow can lead to acute right-side heart failure (Fixler et al., 1973; Vlahakes et al., 1981~. Conversely, a stable model of left ventricular hypertrophy can be produced in 8-week-old pups by aortic banding, which causes a systolic pressure gradient of 15-20 mm Hg (O'Kane et al., 1973~. Dogs with induced tricuspid valve insufficiency can tolerate increased venous pressure and a slight reduction in cardiac output for years, although some develop ascites and reduced serum albumin (Arbulu et al., 1975~. These models require less frequent monitoring. Equipment. Follow-up care and monitoring require appropriate equip

SPECIAL CONSIDERATIONS 87 ment and laboratory support for obtaining CBCs, blood cultures, serum chemistry profiles, and blood-gas analyses. Electrocardiography and echocardiography should be available for assessing cardiac rhythm and function, respectively. Echocardiography is also a useful noninvasive method for monitoring changes in cardiac wall thickness, cardiac motion, and chamber size as cardiac disease progresses. A cardiac catheterization laboratory should be available for performing hemodynamic and angiographic studies. Pharmacologic therapy. Pharmacologic management of dogs that de- velop complications or clinical signs of heart failure must be coordinated between the veterinary unit and the investigator to prevent the administra- tion of medications that could compromise the scientific aims of the study. Diuretics can be used to treat pulmonary edema and reduce plasma volume, but their effects on serum electrolytes and the reduction of venous return and cardiac output should be considered. Vasodilators, calcium antago- nists, ,8-blocking drugs, and positive inotropic agents should be available for managing acute clinical events; however, long-term use of these drugs is usually contraindicated because of their effects on the disease process being studied (Bonagura, 1986; Swindle et al., 1991~. ~ . . . . ~ . . . Hypertension Clinical Features To provide proper care for hypertensive dogs and to avoid inappropri- ate treatment that can be detrimental to the dogs and compromise the study, it is necessary to have a full understanding of the pathophysiology of hyper- tension and of the specific method that is used to induce it. Generally, hypertension in dogs is induced by constricting the renal artery. The result- ing reduction in renal perfusion causes systemic arterial pressure and re- nal arterial pressure distal to the constriction to rise enough to maintain renal function. A discussion of the relationship between renal function and the long-term control of blood Dressure can be found in anv .standarr1 nhv~i- ology textbook (e.g., Guyton, 1991~. ~. . . --, ~r~~~~~ Two methods are most commonly used to induce renal vascular hyper- tension: partial constriction of one renal artery (the 2-kidney, 1-clip method) and unilateral nephrectomy and partial constriction of the remaining renal artery (the 1-kidney, 1-clip method). Both those methods produce what is called Goldblatt hypertension, but the mechanisms responsible for the hy- pertension are different. The 2-kidney, 1-clip model depends more heavily on the renin-angiotensin system than the 1-kidney, 1-clip model and re- sponds to acute treatment with angiotensin-converting enzyme (ACE) in- hibitors, which block the conversion of angiotensin I to angiotensin II. The . . . .

88 DOGS: LABORATORY ANIMAL MANAGEMENT 1-kidney, 1-clip model requires chronic treatment with ACE inhibitors to lower blood pressure. The reason for that difference is described in detail by Guyton (1991~. The greatest success in producing hypertension while reducing the inci- dence of malignant hypertension and renal failure is achieved by reducing renal arterial flow by exactly 50 percent. Renal blood flow is usually measured when the arterial clamp (Goldblatt clamp) is adjusted during sur- gery, this obviates later surgery to readjust the degree of constriction. Methods have been developed for measuring renal blood flow chronically and adjust- ing the renal artery clamp (Ferrario et al., 1971), and more recently a tech- nique has been described for producing hypertension reliably by gradually constricting the renal artery with constrictors fabricated of ameroid, a hydroscopic material made of compressed casein cured in formalin (Ben et al., 1984; Brooks and Fredrickson, 1992~. Other methods that have been used for inducing hypertension include a 2-kidney, 2-clip model in which Goldblatt clamps or ameroid constrictors are applied to both renal arteries; wrapping of one or both kidneys with silk or cellophane; a combination of unilateral nephrectomy and wrapping of one kidney; and placing sutures in a figure 8 configuration on the surface of one or both kidneys (the Grollman model). The creation of hypertension with deoxycorticosterone acetate (DOCA) and common salt has not been as successful in dogs as it has in rats, because dogs are reluctant to eat a high- salt diet or drink a saline solution. However, moderate hypertension in dogs can be achieved with DOCA administration alone. A colony of spon- taneously hypertensive dogs has been described (Bovee et al., 1986~. Husbandry and Veterinary Care Proper care of hypertensive dogs involves the following: · careful design and establishment of the hypertensive model to pro- duce stable hypertension; routine evaluation of renal function; · regular and frequent monitoring of blood pressure · regular monitoring of the retinas; · appropriate treatment with antihypertensives when required; and . careful husbandry. Evaluation of renal function. Routine evaluation of renal function is essential because renal failure is a common complication in dogs with ex- perimental hypertension. Renal failure can be caused by too much constric- tion of the renal artery, a rapid increase in both systolic and diastolic pres- sures (malignant hypertension), or the inappropriate use of antihypertensives.

SPECIAL CONSIDERATIONS 89 Evaluation of renal function is especially important with use of the 1-kid- ney, 1-clip and 2-kidney, 2-clip models (which cause the most severe hy- pertension) and during antihypertensive therapy. In hypertensive dogs, re- nal function is compromised to such an extent that blood pressure must be raised to maintain sodium balance. If antihypertensive therapy lowers blood pressure too much, acute renal failure will ensue. The most reliable and easily measured indicators of renal function are serum creatinine concentration and blood urea nitrogen (BUN). Although they depend somewhat on the type of assay, normal serum creatinine for the dog ranges between 0.4 and 1.3 mg/dL and BUN between 10 and 25 mg/dL. Serum creatinine and BUN should be determined in each dog before hyper- tension is induced to avoid using dogs with already-compromised renal function. In Goldblatt hypertensive models, serum creatinine should be determined daily for the first 5 days after surgery and twice a week thereaf- ter. If ameroid constrictors are used, daily evaluations should continue through the second week after surgery because it takes 4-5 days for ameroid constrictors to reach maximal constriction. In models in which hyperten- sion is not as severe, such as the 2-kidney, 1-clip and 1-kidney, 1-wrapped hypertensive models, renal function is less likely to be impaired, and serum creatinine concentration and BUN might not be increased, but they should be evaluated at least once during the 10-day postoperative period. If renal-function tests show signs of renal failure, corrective action should be taken. Too-severe constriction of the renal artery can be cor- rected surgically, or the study can be terminated by euthanatizing the ani- mal. Renal failure caused by lowering blood pressure to below the renal autoregulatory range should be corrected by reducing the dose of the antihypertensive drug to a point that allows blood pressure to remain high enough to maintain renal function. Malignant hypertension can be treated with antihypertensives and reduced salt intake (Ross, 1989; see below). Measurement of arterial blood pressure. Blood pressure should be determined routinely after surgery. It can be done with indirect methods, such as placing a pressure cuff at the base of the tail (Petersen et al., 1988) or above the hock, or with direct methods, such as chronic implantation of arterial catheters or acute femoral arterial catheterization. To avoid compli- cations associated with exteriorized catheters, some investigators now use methods that do not require exteriorized components, such as a Vascular Access Port (Access Technologies, Skokie, Ill.) (Mann et al., 1987), or chronic instrumentation, such as constriction of the carotid loop (Brooks et al., 1991~. In addition, improved telemetric monitoring (Lange et al., 1991) has the potential to allow continuous monitoring of blood pressure over a number of days or weeks. It is important to establish a baseline blood pressure before inducing

go DOGS: LABORATORY ANIMAL MANAGEMENT hypertension. Measuring blood pressure several times permits the dog to become accustomed to the monitoring technique and thereby avoids in- creases in blood pressure caused by stress. Some investigators measure blood pressure indirectly (e.g., with the tailcuff method) before surgery and use more direct methods later. That is done in recognition that indirect methods can lead to a deviation of up to 10 mm EIg from true arterial pressure. Normal systolic blood pressure ranges from 112 to 142 mm Hg; normal diastolic pressure from 56 to 110 mg Hg. Measurements greater than 160/95 indicate hypertension. Treatment for hypertension. When induced correctly, surgically cre- ated hypertension is sustained and has few complications. If necessary, hypertensive dogs can be maintained on special diets (see below) and given diuretics or other antihypertensive drugs when needed. Some drugs readily available for treatment of hypertensive dogs are listed in Table 6.3. Malignant hypertension must be diagnosed quickly and treated aggres- sively. The most striking clinical sign of malignant hypertension can be blindness caused by retinal detachment, which is usually preceded by reti- nal hemorrhage, dilation of retinal vessels, and subretinal edema. The dogs do not appear to be in pain but often bump into walls and might become disoriented or sit quietly in their pens or cages. Diagnosis can easily be confirmed with an ophthalmologic examination. If blood pressure can be controlled and retinal disinsertion (detachment from the ore ciliaris retinae) _ . TABLE 6.3 Drugs Available for the Oral Treatment of Hypertension in DOgsa Dosage, Frequency of Generic Name mg/kg Administration Class Chlorothiazide 20-40 Every 12 hr or daily Diuretic Hydrochlorothiazide 2-4 Every 12 hr or daily Diuretic Furosemideb 2-4 Every 8-12 hr Diuretic Propranolol 0.25-0.5 Every 8 hr ,6-Adrenergic antagonist Hydralazine 1-3 Every 12 hr Vasodilator Prazosin 0.25-2 Every 8 hr Vasodilator VerapamilC 1-2 Every 8 hr Vasodilator; calcium channel blocker Captopril 0.5-1 Every 8-12 hr Angiotensin-converting enzyme inhibitor aAdapted from Ross, 1989; printed with permission of the author and W. B. Saunders, Philadelphia, Pennsylvania. bCan also be given intramuscularly or intravenously at 2-4 mg/kg. CCan also be given intravenously at 0.05-0.15 mg/kg.

SPECIAL CONSIDERATIONS 91 does not occur, some vision might be restored in 2-3 weeks. Malignant hypertension often responds well to treatment with ACE inhibitors. Diuret- ics can also be administered if care is taken to avoid a precipitous drop in renal blood flow. Vasodilators can be used with caution. If the cause of the malignant hypertension is overconstriction of the renal artery, ACE inhibi- tors can be used to stabilize the dog while the stricture is surgically cor- rected. Husbandry. Routine care of hypertensive dogs must include a consid- eration of diet because both salt intake and protein intake will affect blood pressure and renal function. A high salt intake will exacerbate hyperten- sion, and a high protein intake might accelerate the loss of renal function. To avoid unintended changes in diet that could compromise their dogs and studies, investigators, veterinarians, and others caring for hypertensive dogs should establish dietary requirements before beginning studies. For dogs with hypertension and renal failure, the diet should contain 0.1-0.3 percent sodium on a dry-weight basis or 10-40 mg/kg per day (5-20 mg/lb per day) (Ross, 19899. Low-protein diets (less than 15 percent) that are also low in sodium (e.g., K/D, Hill's Pet Products, Inc., Topeka, Kan- sas) are available and should be fed in adequate amounts-generally 1 can or 2 cups of dry food for each 10 kg (20 lb) of presurgical body weight. The protein content of some commercially available diets might be too low to maintain ideal body weight, but diets that combine a higher protein con- tent with a lower sodium content are available (e.g., R/D, Hill's Pet Prod- ucts, Inc., Topeka, Kansas). As in any dog model, following the body weight of an animal regularly is a good way to monitor the animal's overall health. There is usually no reason to restrict primary enclosure size for hypertensive dogs. Whether they should be exempted from an exercise program depends on their postoperative course. If the hypertensive condition stabilizes and there are no complications, exemption from exercise should not be neces- sary. Blood pressure is known to increase in stressful conditions; therefore, it is important that such conditions be avoided (e.g., dogs that are housed or exercised in pairs or groups should be monitored to ensure that they are compatible). Ehlers-Danlos Syndrome Clinical Features Ehlers-Danlos syndrome type 1 is an autosomal dominant condition of humans for which there are analogues in dogs and other mammals (Hegreberg et al., 1969, 19701. The disease is caused by a defect in metabolism of

92 DOGS: LABORATORY ANIMAL MANAGEMENT dermal collagen that results in a skin tensile strength less than 10 percent of normal. Fibrous tissue and bone are subclinically affected in some cases (Minor et al., 1987~. Multiple lacerations are often observed. The hyperextensible skin can cause superior entropion, inferior ectropion, or both. Husbandry and Veterinary Care The extreme fragility of the skin must be considered in managing dogs with this syndrome. Affected dogs should be housed singly in smooth- surfaced pens of glass, concrete, or sheet steel. Automatic watering valves and other projections should be avoided. The dogs' nails should be kept trimmed. Some dogs might have to wear Elizabethan collars for extended periods to prevent self-inflicted wounds. Dogs should be given opportuni- ties for exercise, either singly or in small groups, by being released under supervision into an exercise pen or room that is free of sharp projections. Leash-walking should be avoided. Wound management is relatively simple. Wounds tend to heal well, possibly because hyperextensible skin places little tension on wounds. Cut- ting suture needles and single-stranded nylon suture material can tear through the skin, but tapered needles and braided sutures, such as those of polygalactin 910 (Vicryl), are well tolerated. It is important to avoid placing too much tension along a single suture line. Hygromas and hematomas, which can become large under loose skin, can be encountered, either as sequelae to lacerations or as primary events. Adhesive tape should never be applied directly to the skin or fur during bandaging because it can tear the skin when the bandage is removed. Entropion and ectropion can be corrected surgically; however, repeated correction might be necessary. Reproduction All affected dogs appear to be heterozygotes; affected homozygotes probably die in utero. To increase fertility, to avoid injury of affected animals, and to prevent conception of homozygotes, it is preferable to select normal bitches and affected males for breeding and to use artificial insemi- nation. Heterozygous affected pups can be identified at birth by the fragil- ity and hyperextensibility of their skin, as can heterozygous fetuses in late gestation.

SPECIAL CONSIDERATIONS Clinical Features 93 Endocrinologic Diseases Endocrinopathies in the dog pose diagnostic and therapeutic challenges because they are complicated physiologic derangements that often involve multiple organ systems. An endocrinopathy might be a desired element of an experimental design or simply a spontaneous random occurrence that would be expected in any canine population. Table 6.4 lists the major endocrinopathies that have been documented in dogs. Discussions in this section are limited to endocrinopathies that either are induced in experimen- tal animals or are undesired results of management procedures or investiga- tional protocols. Hypothyroidism and hyperadrenocorticism (Cushing's disease), two major endocrinopathies often seen in clinical veterinary practice, are not discussed here but are well described in the veterinary medical literature (e.g., Capen and Martin, 1989; Chester, 1987, Drazner, 1987a; Feldman, 1989; Hsu and Crump, 1989; Peterson and Ferguson, 1989~. A brief review of disorders of calcium metabolism is included because hypocalcemia caused by iatrogenic hypoparathyroidism occasionally occurs in a research setting, and hypercalcemia is often mistakenly attributed to parathyroid dysfunc- tion. TABLE 6.4 Selected Endocrine Disorders in Dogs Affected Organ Diseases Adrenal cortex Adrenal medulla Pancreas Parathyroid Pituitary Thyroid Multiple glands Hyperadrenocorticism H. . . ypoaarenocort~c~sm Pheochromocytoma Diabetes mellitus Gastrinoma Hyperparathyroidism Hypoparathyroidism Acromegaly Diabetes insipidus Hypopituitarism Hyperthyroidism Hypothyroidism Hyperlipidemia Hypoglycemia

94 DOGS: LABORATORY ANIMAL MANAGEMENT TABLE 6.5 Common Clinical Signs of Selected Canine Endocrinopathies Endocrinopathy Common Clinical Signs Diabetes mellitus Hyperglycemia, polydipsia, polyuria, glycosuria, increased food consumption but loss of weight, bilateral cataract develop ment, weakness Hypoadrenocorticism Weakness, vomiting, diarrhea, bradycardia, acute collapse Acromegaly Respiratory strider, increased interdental spaces, prominent skin folds, abdominal enlargement, fatigue Hypercalcemia Mental dullness; muscular weakness; tachycardia; upper gastrointestinal signs, including anorexia, nausea, and vomiting; signs of renal disease, including nephrocalcinosis, renal calculi, and secondary renal failure Hypocalcemia Muscle tremors, tetany, seizures Common clinical signs of the endocrinopathies to be discussed are listed in Table 6.5. They range from very subtle changes to acute crises. Most are nonspecific and can also be seen in various nonendocrine disorders. Detailed discussions of endocrinopathies can be found in the veterinary medical literature (e.g., Drazner, 1987b; Ettinger, 1989; Feldman and Nelson, 1987; McDonald and Pineda, 1989; Morgan, 1992~. Husbandry and Veterinary Care Procedures for managing dogs with endocrinopathies are dictated by both the experimental design and the animals' welfare. Diabetes mellitus. Diabetes mellitus in the dog is a recognized sponta- neously occurring model (Kramer, 1981), and the disease is readily induced either by chemical ablation of the pancreatic p-cells or by total pancreatectomy (Mordes and Rossini, 1985~. Frequent monitoring is mandatory for the successful management of dogs with diabetes mellitus. Daily measure- ments, before the first meal of the day and 6-12 hours later, are required to stabilize and control blood glucose in diabetic dogs. The second glucose measurement can be eliminated only when the afternoon blood glucose of an individual dog is consistent from day to day and the insulin requirement for that dog is well established. Blood glucose monitoring should begin after initial administration of diabetogenic chemicals or during the first 24 hours after pancreatectomy. Fasting blood glucose, as measured by the plasma or serum glucose oxidase method, ranges from 65-118 mg/dL (3.6- 6.5 mmol/L) in normal adult dogs (Kaneko, 1989~. A number of insulin preparations can be used either singly or in combi- nation in dogs: regular, NPH, lente, and ultralente. Unit doses and prepara

SPECIAL CONSIDERATIONS 95 lion types must be determined for and adjusted to the response of each dog. Insulin should be started at a dose of 1 U/kg per day injected subcutane- ously at the time of feeding the first meal of the day. Daily proportions of each preparation included in a therapeutic regimen are determined by trial and error as guided by the results of serial blood glucose measurements. Detailed information on dosage and characteristics of various insulin prepa- rations is available (Nelson, 1989; Schaer, 19921. In addition to insulin administration, stresses from environmental and experimental manipulation, exercise, concurrent disease, estrus, and changes in food and water intake can cause profound fluctuations in blood glucose concentrations. Blood glucose can be manipulated by adjusting insulin types and dosages. As a general rule, it is preferable to have a slightly hyperglycemic dog rather than a hypoglycemic one because of the potentially disastrous results of a hypoglycemic crisis. If such a crisis occurs, it should be treated with intravenous dextrose and supportive care (Kirk and Bistner, 1985~. Supplemental glucose can be given orally if the dog is able to swallow. Obviously, a necessary follow-up includes reviewing and adjusting the in- sulin dosage and the ratio of short- to long-acting insulin preparations given. The amount of food fed to each diabetic dog should be standardized at what is necessary to maintain its optimal body weight. The same amount should be fed each day. Once an eating pattern (amount of food eaten and time required for meal consumption) is established for a given dog, its appetite can be used as an indicator of general well-being. In pancreatectomized dogs, it is necessary to compensate for lost pan- creatic exocrine function. That can be accomplished by adding a commer- cially available digestive enzyme to the food. Some dogs find the product unpalatable, but it is generally accepted if it is mixed with canned food. Diabetic dogs can be maintained for long periods, but sequelae of dia- betes mellitus including neuropathy, immune system compromise, and de- layed healing do occur, and a shorter than normal life span should be expected. Hypoadrenocorticism. The canine model of hypoadrenocorticism (Addison's disease) is a classic model in biomedical research (Brown-Sequard, 1856~. Hypoadrenocorticism can be induced in dogs by administering the drug mitotane,1 which chemically ablates the adrenal cortex (Nelson and Woodard, 1949~. During induction, a presumptive diagnosis can be made by monitor- ing changes in serum electrolytes, specifically sodium and potassium. The normal ranges of sodium and potassium concentrations in dog serum are 1 Chemical name, 1,1-dichloro-2-(o-chlorophenyl)-2-(p-chlorophenyl) ethane; trivial name, o,p'-DDD; brand name, Lysodren.

96 DOGS: LABORATORY ANIMAL MANAGEMENT 140-155 mEq/L and 3.7-5.8 mEq/L, respectively (Carlson, 1989~. In dogs with hypoadrenocorticism, the sodium-to-potassium ratio is decreased to less than 27:1 (Schrader, 1988), although this hyperkalemia is not pathognomonic. The adrenal corticotropic hormone stimulation test is required for definitive diagnosis (Nichols and Peterson, 1992~. In a crisis, resuscitation requires recognizing the problem, intravenously administering 0.9 percent saline so- lution, replacing glucocorticoids and mineralocorticoids, and possibly pro- viding therapy for hyperkalemia. Long-term maintenance entails glucocor- ticoid (cortisone) administration, mineralocorticoid supplementation with 9-fluorohydrocortisone acetate,2 and the addition of sodium chloride to the diet. Electrolytes should be monitored at least weekly once stabilization is achieved. Environmental and experimental stresses and alterations in water and food availability can have substantial effects on electrolyte balance and homeostasis. Additional glucocorticoid (increased by a factor of 2-1OJ should be administered during periods of stress. Acromegaly. Acromegaly can be iatrogenically induced in bitches when progesterone is given to prevent estrous cycling (Eigenmann, 1985, 19891. It can also be secondary to increased production of progesterone during diestrus. Progesterone induces acromegaly by increasing the production of growth hormone in the anterior pituitary gland. The excessive release of growth hormone can also induce a "pituitary diabetes" that can be difficult to control with insulin. Cessation of progesterone administration or spaying will reverse acromegalic changes. Calcium derangements. Although disorders of the parathyroid glands are usually suspected when hypercalcemia or hypocalcemia is present, the calcium abnormality is more often associated with other conditions, includ- ing pseudohyperparathyroidism, the most common cause of hypercalcemia (Feldman and Nelson, 1987~; hypoadrenocorticism; renal failure; bone le- sions; and hypervitaminosis D. Primary hyperparathyroidism in the dog is rare. Pseudohyperparathyroidism (hypercalcemia of malignancy) is a paraneoplastic syndrome that has been recognized in dogs with lymphosar- coma, adenocarcinoma of the anal apocrine glands, multiple myeloma, osteosarcoma, and other neoplasms (Meuten et al., 1982, 1986~. Signs of hypercalcemia are not always overt, and treatment should be directed to- ward the underlying cause. Causes of hypocalcemia include calcium imbalance during lactation, renal disease, acute pancreatitis, intestinal malabsorption, hypoalbuminemia, and primary hypoparathyroidism (idiopathic or iatrogenic). Iatrogenic 2 Brand name, Florinef.

SPECIAL CONSIDERATIONS 97 hypoparathyroidism is associated with inadvertent damage or removal of the parathyroid glands and is an important consideration in research set- tings. Surgery involving the ventral neck area or the laryngeal-tracheal area or removal of the thyroid glands carries an increased risk of complications related to parathyroid function. Treatment includes calcium replacement and appropriate management of the precipitating disorder. Hematologic Disorders Clinical Features Canine models of human hematologic disorders have been reviewed (Dodds, 1988, 1989, 1992; Hall and Giger, 1992; Harvey, 1989; Kaneko, 1987; Knoll, 1992~. Clinical signs of some of these disorders are listed in Table 6.6. TABLE 6.6 Inheritance and Signs of Selected Hematologic Disorders in Dogs Disorder Inheritance Clinical signs Hemophilia A X-linked Hemophilia B X-linked (Christmas disease) Low factor VIII coagulant activity but normal or increased von Willebrand factor antigen concentrations; spontaneous bleeding diathesis of varied severity, depending on factor VIII activity; severely affected dogs often exhibit spontaneous hemarthroses and large joints. The most common severe inherited bleeding disease. Recognized in most purebreds and in mongrels. Deficiency of factor IX activity; signs similar to those of hemophilia A. Recognized in 17 breeds. von Willebrand's Autosomal Variable deficiency of von Willebrand factor; disease type I incompletely factor VIII activity might be reduced; and dominant prolonged bleeding time; moderately severe bleeding diathesis of mucosal surfaces. Signs are exacerbated by stress, hypothyroidism, intercurrent disease, trauma, and surgery. Recognized in more than 50 breeds. continued on next page

98 TABLE 6.6 Continued DOGS: LABORATORY ANIMAL MANAGEMENT DisorderInheritance Clinical signs von Willebrand'sAutosomal disease type IIIrecessive Factor X deficiency Autosomal incompletely dominant Thrombopathia Autosomal Cyclic hematopoiesis Autosomal Pyruvate kinase Autosomal deficiency Erythrocyte recessive phosphofructokinase recessive deficiency Severe deficiency of von Willebrand factor; factor VIII activity is usually low; indefinitely prolonged bleeding time; mucosal surface bleeding diathesis, which can be severe and is exacerbated by stress, hypothyroidism, trauma, surgery, and intercurrent disease. Recognized in Chesapeake Bay retrievers, Scottish terriers, and Shetland sheepdogs. Homozygotes are stillborn or die shortly after birth; affected pups might live for up to 2 weeks and then die of massive internal bleeding; young adults can also exhibit life- threatening hemorrhage, but signs in mature adults are usually mild and confined to mucosal surfaces. Found only in one large family of cocker spaniels. Affected dogs can have no clinical signs or show increased bleeding tendency that can be exacerbated by trauma or surgery. Found in basset hounds and otterhounds. Regularly occurring interruptions of bone marrow hematopoiesis with loss of neutrophils from peripheral blood; during these periods, dogs exhibit fever, enteritis, keratitis, pneumonia, and skin infections; infections can become life-threatening if not treated. Found in gray collies. Affected dogs exhibit severe anemia with reticulocytosis, macrocytosis, and p o ly c hro m as i a; hyperb ilirubinemi a ; splenomegaly with extramedullary hematopoiesis; and decreased red cell survival. Found in basenjis, beagles, and cairn terriers. Autosomal Persistent compensated hemolytic anemia with episodes of intravascular hemolysis, hemoglobinuria, and fever associated with stress or exercise; hemolytic crises follow hyperventilation-induced alkalemia; red cells of affected dogs are extremely alkaline and fragile in vitro. Found in English springer spaniels.

SPECIAL CONSIDERATIONS Husbandry and Veterinary Care 99 Bleeding disorders. Dogs with congenital and acquired bleeding disor- ders require special housing to minimize the risk of spontaneous or injury- induced bleeding. This is important not only for the animals' welfare, but also for experimental reasons. The basal state of animals that experience repetitive bleeding can be altered by the physiologic stress that such bleed- ing causes and, if bleeding is severe enough to require transfusions, by repeated exposure to homologous plasma proteins and blood cells. That is of particular concern for dogs with severe disorders, such as hemophilia. Dogs with bleeding disorders should be housed in enclosures that have smooth sides and fronts with smooth vertical or cross-hatched bars. It is not advisable to use materials that can be climbed (e.g., chain-link fencing) because dogs with bleeding disorders can suffer foot injuries caused by weight-bearing pressure between the toes. Enclosure size is also important. To prevent injury, affected animals should have sufficient space to move about freely but not enough to permit vigorous exercise if they become excited. Enclosures should be square or oblong; injury is more likely to occur in a long, narrow run, especially in dogs with long tails, which during wagging can be traumatized by hitting against the sides. Experience has shown that for dogs weighing from 13.6-36.3 kg (30-80 lb), primary hous- ing measuring about 4 x 6 ft (1.22 x 1.83 m) or 5 x 5 It (1.52 x 1.52 m) minimizes the risk of injury. Severely affected dogs should be housed individually because the risk of injury in playing with other dogs is substantial. To provide socialization, it is advisable to construct pens that allow visual contact between dogs; this can be achieved by building pens across an aisle from or perpendicular to each other. Partitions between the runs should be solid for the first 4 It (1.22 m) in height to prevent injury caused by dogs in adjacent pens playing or fighting through the partition, and the seam with the floor should be smooth. To avoid foot-pad abrasions, nonslip flooring should not be too rough. A poured rubberized flooring with a small amount of sand added to the last coat should create enough friction to prevent sliding. Nontoxic bedding (e.g., shredded newspaper or shavings) can be used to minimize injuries if sliding does occur. English rubber colts or tennis balls can be used to provide environmental enrichment. Special arrangements are required for feeding and watering. Automatic watering devices are generally not recommended because the spigots can cause mouth injuries, and bleeding from such injuries is usually difficult to control. It is better to use large water buckets anchored to the sides or fronts of the pens. Dry food should be softened before feeding and supple

100 DOGS. LABORATORY ANIMAL MANAGEMENT mented with good-quality canned or cooked meat. A hematinic can be added to the food for conditioning. Hard biscuits should not be fed. Bleeding from small surface injuries to the gums or nose or from toe- nails that are cut too short can be stopped by using sealant materials, such as Nexaband glue (Tri-Point Medical, LP, Raleigh, N.C.~. Bleeding toe- nails can also be packed with styptic powder, and the soft rubber end cap from an intravenous set or catheter can be wedged tightly over the nail. If necessary, the foot can be bandaged; this supplies enough local pressure to control the bleeding. For animals that experience severe bleeding episodes, transfusion is the treatment of choice. Fresh-frozen plasma, plasma concen- trates, platelet concentrates, or packed red cells should be given as required for the specific disorder. Details of management and treatment are summa- rized elsewhere (Dodds, 1989, 1992~. Another management procedure to keep animals healthy and reduce bleeding risk is prophylactic dentistry, which must be performed very care- fully to avoid injury to the gums. Booster vaccinations should not be given during bleeding episodes because they create a transient platelet deficit (Dodds, 1992~. In addition, dogs are at increased risk for bleeding episodes for 10-14 days after vaccinations. Affected females sometimes bleed exces- sively both during estrus and during the 30-40 days beforehand when estro- gen concentrations are elevated. Cyclic hematopoiesis. Colonies of grey collies with cyclic hematopoie- sis (formerly called cyclic neutropenia) have special requirements because they are susceptible to recurring infections and anemia (Knoll, 1992~. They have a cyclic, profound drop in all their blood-cell classes, although the numbers of each cell type rise and fall at different times. Affected animals rarely live beyond the age of 3 years and experience frequent bleeding episodes from cyclic thrombocytopenia. Respiratory tract and enteric infec- tions are the most debilitating. Affected animals can often be housed together, but they need scrupu- lously clean facilities to minimize infection, close clinical monitoring, and supportive therapy. They should be monitored for neutropenia, and prophy- lactic antibiotics should be administered as neutrophil counts begin to de- cline. Other hematologic disorders. Dogs with various other inherited and acquired hematologic diseases also require special care. For example, basenjis with pyruvate kinase deficiency and recurring anemia must be closely monitored because of their increased susceptibility to infection or stress (Hall and Giger, 1992; Harvey, 19891; beagles with hereditary nonspherocytic hemolytic anemia must be closely monitored for episodes of hemolytic crisis (Maggio- Price et al., 1988~; and English springer spaniels with erythrocyte phospho

SPECIAL CONSIDERATIONS 101 fructokinase deficiency require special care during episodes of hemoglobinuria or myoglobinuria (Hall and Giger, 1992; Harvey, 1989~. Reproduction For dogs with severe inherited bleeding disorders such as hemophilia, von Willebrand's disease, factor X deficiency, and platelet dysfunction (thrombopathia)- special care is needed for breeding, whelping, and rear- ing of the offspring. Immediately after birth, each pup should be carefully examined for signs of bleeding, its umbilical cord should be ligated, and the potential for trauma from the dam should be minimized. It might be neces- sary to tranquilize first-time dams slightly to protect the young. When the pups are weaned and start to become more active, blood samples should be taken to determine which pups are affected. In hemophilia, the affected pups from a carrier (heterozygous) dam will be males, unless the sire is a hemophiliac (hemizygote), in which case both affected hemizygote males and homozygote females can be produced. Generally, male pups should be watched more closely, and the affected ones should be removed and housed separately if the litter is too rambunctious. Cages should be relatively small; a floor area of about 30 x 36 in (76 x 91 cm) is recommended for the average hemophilic pup. Affected pups should be watched carefully after vaccinations. Modi- fied live-virus vaccines might induce a relative thrombocytopenia and plate- let dysfunction during the period of viremia (i.e., 3-10 days after vaccina- tion) (Dodds, 19923. The pups are at substantial risk for spontaneous or traumatic bleeding at this time because the vaccine effect on platelet func- tion superimposes another hemostatic burden. All vaccinations should be given subcutaneously with a small-gauge needle, preferably 23 or 25 gauge, in the loose skin folds of the neck. Intramuscular injections in affected animals should be avoided. Affected pups should be housed initially in cages and eventually in small pens. At teething, affected puppies often bleed excessively from the gums; this necessitates use of a topically applied sealant and, on occasion, transfusion therapy. Immunologic Diseases Primary Immunodeficiency and Autoimmune Diseases Clinical Features Immunodeficiency is characterized by failure to manifest a normal im- mune response when challenged by infectious agents or other substances

102 DOGS: LABORATORY ANIMAL MANAGEMENT that are foreign to the body. The subnormal response can result from a defect in the afferent, central, or efferent limb of the immune system (see review in NRC, 1989~. Immunodeficiency disorders can be primary (i.e., inherited) or secondary (i.e., acquired). Primary immune deficiency can result from an inherited defect in immunocompetent cells or effecter mechan- isms (e.g., complement or phagocytes) or can be associated with autoim- mune disease or a deficiency in growth factors necessary for the optimal function of immunocompetent cells (WHO Scientific Group, 1986~. Sec- ondary immune deficiency can be caused by various environmental factors, including x rays, viral agents, toxic chemicals, and dietary deficiencies. Several primary immunodeficiency diseases have been described in dogs, including selective IgA deficiency (Campbell, 1991; Felsburg et al., 1985; Moroff et al., 1986), IgM deficiency (Mill and Campbell, 1992; Plechner, 1979), common variable immunodeficiency (A. Rivas, New York State Col- lege of Veterinary Medicine, Cornell University, Ithaca, N.Y., unpublished), and severe combined immunodeficiency disease (Jezyk et al., 1989; Patterson et al., 1982~. Dogs with particular autoimmune diseases also suffer from immunodeficiency. A high incidence of septicemia has been observed in dogs that were bred to develop systemic lupus erythematosus (SLE) (Quimby et al., 1979~. Autoimmune hemolytic anemia (AlIA) (Bull et al., 1971; Dodds, 1983; Klag et al., 1993), immune thrombocytopenic purpura (ITP) (Dodds, 1983, 1992; Waye, 1960), SLE (Grindem and Johnson, 1983; Monier et al., 1988; Quimby, 1981), rheumatoid arthritis (RA) (Bell et al., 1991; Carter et al., 1989; Quimby et al., 1978), Sjogren's syndrome (Kaswan et al., 1985; Quimby et al., 1979), autoimmune thyroiditis (Gosselin et al., 1982; Quimby et al., 1979; Rajatanavin et al., 1989; Thacker et al., 1992), and thyrogastric disease (Quimby et al., 1978) have been found in research dogs. Primary immunodeficiencies in dogs have also been associated with the absence of the third component of complement (Winkelstein et al., 1981~; deficits in neutrophil function, including cyclic hematopoiesis (see page 100) (Knoll, 1992; Lund et al., 1967) and granulocytopathy (Knoll, 1992; Renshaw and Davis, 1979~; dysregulation of interleukin-6 (DiBartola et al., 1990; Rivas et al., 1992~; and deficiency of growth hormone (Roth et al., 1980~. Clinical signs of these diseases are presented in Table 6.7. All dogs with primary immunodeficiencies are predisposed to infection. Dogs with disorders associated primarily with hypogammmaglobulinemia, complement, or phagocytic function are predisposed to bacterial infection (Blum et al., 1985; Lund et al., 1967; Moroff et al., 1986; Renshaw and Davis, 1979~. Those with disorders of cell-mediated immunity have in- creased susceptibility to fungi and viruses (Jezyk et al., 1989~.

SPECIAL CONSIDERATIONS TABLE 6.7 Clinical Signs of Selected Primary Immunodeficiency and Autoimmune Diseases in Dogs 103 Immunologic Disease Clinical Signs Common variable immunodeficiency IgM deficiency Selective IgA deficiency Severe combined immunodeficiency Autoimmune hemolytic anemia Immune thrombocytopenic purpura Systemic lupus erythematosus Rheumatoid arthritis Sjogren's syndrome Autoimmune (lymphocytic) thyroiditis Thyrogastric disease Granulocytopathy Dysregulation of interleukin-6 Deficiency of growth hormone Increased susceptibility to infectious diseases; clinical presentation after the age of 6 months Increased susceptibility to bacterial diseases Increased susceptibility of some dogs to infectious diseases of mucosal surfaces, such as those of gastrointestinal, respiratory, and urogenital tracts Extreme susceptibility to bacterial, viral, and fungal infections; clinical presentation in first few weeks of life; death before reaching maturity Pallor, slight jaundice, splenomegaly, lymphadenopathy, weakness, and shortness of breath; profound anemia and recurrent episodes of hemolytic disease in approximately 50% of affected dogs Bruise easily, prolonged bleeding after trauma Rash, hemolytic anemia, immunothrombo- cytopenic purpura, polyarthritis, and proteinuria; females affected more frequently than males Swollen painful joints generally multiple small . . . art~cu. ar Joints Keratoconjunctivitis sicca (dry eyes); corneal ulcers associated with dry eyes; excessive dental caries; inflamed gums; signs associated with hypothyroidism, including tendency to obesity, tendency to seek warm places, bilaterally symmetrical hair loss, and changes in skin thickness Signs associated with hypothyroidism, including tendency to obesity, tendency to seek warm places, bilaterally symmetrical hair loss, and changes in skin thickness Signs associated with hypothyroidism, inappe- tence, megaloblastic anemia, and atrophic gastritis Increased susceptibility to bacterial infections Familial Mediterranean fever, characterized by fever, synovitis, and renal failure Small body stature; generalized increase in susceptibility to infectious diseases

104 Husbandry and Veterinary Care DOGS: LABORATORY ANIMAL MANAGEMENT Immunodeficient dogs pose special management problems. Immune diseases must be diagnosed, their prognosis determined, and their therapy monitored. A number of tests have been developed for those purposes, including tests that assay T- and B-cell function (Ladiges et al., 1988, 1989), identify serologic markers of autoimmune diseases (Kaplan and Quimby, 1983; Quimby et al., 1980), identify circulating immune complexes in rheu- matic and neoplastic diseases (Carter et al., 1989; Terman et al., 1979), and assay phagocyte function (Smith and Lumsden, 1983~. The susceptibility of immunodeficient dogs to infectious diseases is handled in various ways. All immunodeficient dogs can benefit from an environment that minimizes contact with canine pathogens; however, for some of these conditions (e.g., severe combined immunodeficiency), cesar- ean derivation and maintenance in a gnotobiotic chamber are required to ensure survival. Pups with humoral deficiencies born to normal dams profit from receiving maternal antibodies in colostrum, and their dams should be immunized before being bred to ensure that high concentrations of antibod- ies will be present. Adult dogs with humoral deficiencies can be helped by transfusions of normal or hyperimmune serum or plasma or by administra- tion of purified gamma globulin. Some dogs that are genetically predis- posed to autoimmune diseases can be spared clinical illness for years by housing them in gnotobiotic chambers; however, if they are moved to a conventional environment, they quickly develop autoimmune disease (Schwartz et al., 1978~. Dogs with autoimmune diseases should be carefully monitored and ap- propriately treated. Treatment might involve immunosuppressive therapy (e.g., for SLE, AHA, ITP, and RA), transfusions of red cells and platelets (for AHA and ITP), splenectomy (for AHA and ITP), renal dialysis (for SLE), administration of thyroxine (for autoimmune thyroiditis), administra- tion of thyroxine and vitamin B~2 (for thyrogastric disease), and administra- tion of artificial tears (see the section on ophthalmologic disorders) and special dental care (for Sjogren's syndrome). Some dogs with growth hor- mone deficiency benefit from injections of thymosin (Roth et al., 1980~. Bone marrow transplantation and systemic antibiotics are effective in treat- ing dogs with neutrophil defects. Dogs with thrombocytopenia (as in SLE, ITP, or Evan's syndrome) are predisposed to bleeding and bruising and should be housed and maintained as described in the section on hematologic disorders. Preliminary studies suggest that oral levamisole therapy is effi- cacious in treating one type of canine common variable immunodeficiency that is associated with ulcerative colitis and a predisposition to adenocarcinoma of the intestine (A. Rivas, New York State College of Veterinary Medicine, Cornell University, Ithaca, N.Y., unpublished). Trials involving the use of colchicine to delay the onset of amyloidosis in dogs with interleukin-6

SPECIAL CONSIDERATIONS 105 dysregulation are in progress (L. Tintle, Wurtsboro Veterinary Hospital, Wurtsboro, N.Y., unpublished). The care of dogs with C3 deficiency and dogs that have been exposed to total body irradiation and immunosuppres- sive drugs associated with organ transplantation is described below. Dogs should be immunized against known canine pathogens before being exposed to agents that will induce immunodeficiency. Reproduction In colonies where the objective is to reproduce dogs with SLE by se- lecting breeders with serologic evidence of the disorder (i.e., by using anti- nuclear antibody and LE-cell tests), many progeny develop autoimmune diseases not apparent in the parents (Monier et al., 1988; Quimby et al., 19793. That observation has led to the hypothesis that multiple genes con- trol the susceptibility and specificity of autoimmune diseases (Monier et al., 1988; Quimby and Schwartz, 1978~. In some cases, an unanticipated result is compromised fertility (e.g., immune-mediated aspermatogenesis), which necessitates the use of littermates or repeat breeding of the parents to con- tinue the lineage (Quimby et al., 1978~. Hypothyroidism caused by lym- phocytic thyroiditis (Beierwaltes and Nishiyama, 1968; Gosselin et al., 1982; Mizejewski et al., 1971; Rajatanavin et al., 1989; Thacker et al., 1992) can lead to poor reproductive performance that can be corrected with thyroxine- replacement therapy. Details on monitoring blood thyroxine and oral supple- mentation have been published (DePaolo and Masoro, 1989; Ferguson, 1986~. For some autoimmune diseases, such as immune-mediated aspermatogenesis, no therapy has been found. Complemerlt Deficiency Clin~cal Features Dogs deficient in the third component of complement (C3) are particu- larly susceptible to bacterial infections (Blum et al., 1985~. They also develop a membranoproliferative glomerulonephritis, which can be detected histologically by the age of 1 year (Cork et al., 19911. Affected dogs are normally active and appear well; the only clinical sign of this renal disease is proteinuria. Renal disease progresses inexorably and culminates in a nephrotic syndrome with azotemia when the dogs are 6-8 years old. Husbandry and Veterinary Care Dogs deficient in C3 can be reared and housed in standard laboratory dog facilities. Because the dogs are susceptible to bacterial infections (Chick et al., 1984), animal technicians should be alert to any deviations from

106 DOGS: LABORATORY ANIMAL MANAGEMENT normal behavior that might indicate illness (e.g., inappetence and lethargy). C3-deficient dogs that show these clinical signs must immediately be evalu- ated for increased body temperature and leukocytosis. Blood samples should be taken and submitted for culturing to identify and determine the antibiotic sensitivity of the microorganisms; however, treatment with intravenous bac- tericidal antibiotics should not await diagnosis but should begin as soon as clinical signs are detected and a blood sample has been drawn. Although that protocol undoubtedly results in overtreating and might preclude a de- finitive diagnosis, it will in most cases ensure the recovery and survival of the affected dog. If an invasive procedure (e.g., renal biopsy or placement of an indwelling catheter) is required, antibiotic prophylaxis should begin 24 hours beforehand, and it is essential to follow strict aseptic technique while performing the procedure. The presence of proteinuria can be detected by testing for total-protein excretion in the urine over a 24-hour period, and renal biopsies can be used to evaluate the progression of renal disease. As dogs age, periodic mea- surements of total serum protein, albumin, and serum urea nitrogen can be used to identify dogs whose renal disease is becoming severe or those in which a nephrotic syndrome might lead to fluid accumulation in body cavi- ties. Repeated blood transfusions or infusions of canine plasma are contraindi- cated because they exacerbate renal disease. Reproduction C3 deficiency is inherited as an autosomal recessive trait (Johnson et al., 1986; Winkelstein et al., 19821. Affected pups are produced by breed- ing heterozygous females with homozygous males. Homozygous females are fertile but have rarely produced viable young. Pups should be tested at birth, and the ones that are C3-deficient should be placed on antibiotic therapy for the first 4 days after birth. C3-deficient dogs do not respond normally to immunization; therefore, it is recommended that immunizations against the common canine pathogens be given at 2-week intervals until the pups are 18 weeks old (Krakowka et al., 1987; O 'Neil et al., 1988; Winkelstein et al., 1986~. Organ Transplantation Clinical Features Dogs that are used in organ-transplantation studies must first be made immunodeficient. Immunosuppressive methods include total-body irradia- tion and administration of cytologic chemicals (Ladiges et al., 1989~. Im

SPECIAL CONSIDERATIONS 107 munosuppressed dogs are very susceptible to infectious diseases and might have gastrointestinal tract problems. Husbandry and Veterinary Care Dogs that undergo experimental organ transplantation generally require intensive postoperative supportive care, the level of which depends on the transplantation procedure used and the degree of immunosuppression re- quired to overcome graft rejection. Supportive care includes fluid therapy, blood and platelet transfusions, preoperative and postoperative administra- tion of appropriate antibiotics, and intensive husbandry practices. Regular monitoring of white cells is critical for ascertaining health status and deter- mining the necessity for treatment. Blood should be cultured if clinical signs suggest septicemia. Nutritional needs are critical for dogs undergoing bowel transplantation or for those suffering from gastrointestinal tract prob- lems caused by the immunosuppressive procedures. Dogs might need to be housed individually in intensive-care facilities during early convalescence. Dogs undergoing bone marrow transplantation are profoundly immuno- deficient for 200-300 days after lethal total-body irradiation and successful marrow engraftment, and they require intensive supportive care (Ladiges et al., 1990~. Recovery of granulocyte count and function is complete by the twenty-fifth day after engraftment; blood lymphocyte count does not return to normal until day 200. Antibody response to bacteriophage and sheep and chicken red cells is lower than normal during the first 200 days, with IgM being the primary isotype. Lymphocyte stimulation by phytohemaggluti- nin, the mixed-leukocyte reaction, and the response to first- and second-set skin grafts are impaired. Long-term survivors (dogs that survive more than 200 days' generally regain their health and are no longer more susceptible than normal to infectious diseases. The development of graft-versus-host disease and its treatment drastically affect recovery of the immune system and place the dogs at increased risk for contracting infections. Lysosomal Storage Diseases Clinical Features Clinical manifestations of canine lysosomal storage diseases (LSDs) generally fall into three categories: severe necrologic signs, mainly skeletal signs, and a mixture of visceral, skeletal, and necrologic signs. The follow- ing discussion addresses techniques for managing dogs in each category, using a single LSD as an example. The techniques can be extended to manage dogs with other LSDs.

108 DOGS: LABORATORY ANIMAL MANAGEMENT Fucosidosis. Fucosidosis is caused by a deficiency of oc-~-fucosidase (Healy et al., 1984~. Affected dogs exhibit mainly necrologic signs. By the age of 12 months, affected dogs show subtle behavioral changes and might have an overextended posture. From 12 to 18 months, they develop mild ataxia and hypermetria. Signs progress rapidly between the ages of 18 and 24 months to more severe deficits in gait, proprioceptive defects, hyperclonus, nystagmus, kyphosis, and a loss of learned behavior. The dogs become dull and unresponsive. Hearing and vision might be impaired. Signs in severely affected, 24- to 36-month-old dogs include severe incoordination, opisthotonos, muscle spasms, muscle wasting, circling, head tilt, abnormal papillary light reflexes, dysphasia, and cranial nerve deficits. The dogs become severely obtunded and suffer from self-inflicted injury. If not euthanatized, they usually die by the age of 3 years. Mucopolysaccharidosis VII. The majority of clinical signs in canine mucopolysaccharidosis VII (MPS VII), a condition caused by a deficiency of p-glucuronidase, are related to skeletal and joint abnormalities (Haskins et al., 1984~. Progressive noninflammatory arthrosis develops, and joints become lax and deformed. By the age of 3-6 months, affected dogs are unable to stand, and the muscles of locomotion atrophy. Corneal clouding can lead to decreased vision in dogs with MPS VII, but the impairment is generally less severe than in dogs with MPS I. At the age of 15-22 months, MPS VII-affected dogs often become dull and lethargic and lose interest in their environment and in animal-care personnel. Those signs might be asso- ciated with progressive hydrocephalus. Mucopolysaccharidosis I. Canine mucopolysaccharidosis I (MPS A, a condition caused by a deficiency of oc-~-iduronidase, is most similar to the human MPS I phenotype of intermediate severity (Hurler's syndrome and Scheie's syndrome) (Shull et al., 1982~. Clinical signs refer to visceral, skeletal, and mild necrologic injury. Dogs with MPS I appear normal at birth, although there is a higher than normal incidence of umbilical hernias. Affected pups remain generally healthy for 4-6 months and then show stunted growth, corneal clouding, and progressive, degenerative, noninflammatory joint disease caused by mucopolysaccharide deposition in synovial and periarticular tissues. Joint laxity caused by abnormalities in ligaments and tendons is also common and, in combination with the arthroses, causes decreased ambulation. Degeneration of intervertebral disks, collapse of disk spaces, vertebral and long-bone osteopenia, and spondylosis also de- velop. Mucopolysaccharide accumulation in heart valves and coronary ar- teries can cause rapidly progressing heart failure. Affected dogs remain alert and responsive until their death by natural causes or euthanasia, often between the ages of 2 and 3 years.

SPECIAL CONSIDERATIONS Husbandry and Veterinary Care 109 Dogs with LSD present unique and serious medical and husbandry problems. Proper care of these valuable, critically ill animal models requires compas- sion, diligence, hard work, and specialized knowledge of the diseases in- volved. Technicians must be well trained and observant. Fucosidosis. As the clinical signs progress, affected dogs should be handled carefully to prevent injury. They should be fed, exercised, and housed separately from normal dogs. Severely ill dogs should be moved by carrying. Affected dogs should always be housed on a raised trampoline bed and kept dry during cage cleaning to prevent self-soiling and pressure sores. Particular attention should be given to dogs with long hair; they should be bathed weekly and clipped several times a year. Ears should be checked daily for signs of infection. Dogs with moderate to advanced disease should be fed more frequently, and canned or moistened dry food should be used to aid prehension. Dogs with advanced disease often have a poor appetite, and the addition of highly palatable foods assists in maintain- ing body weight. Excess dental tartar must be removed regularly. At the age of 2-3 years, motor and mental impairment will have progressed to the point that euthanasia will be indicated. Mucopolysacchar~dosis VII. MPS VII-affected dogs should be housed in cages with floors of coated wire mesh; this aids sanitation and helps to prevent decubital sores. Once the dogs are unable to walk, food and water intake must be carefully managed. Recumbent animals will usually eat and drink if pans of food and water are placed on the cage floor; however, hand feeding might become necessary. Euthanasia should be considered when a dog's response to human attention begins to diminish. Mucopolysaccharidosis I. Except for surgical correction of umbilical hernias, special care is not usually required for dogs with MPS I that are less than 1 year old. However, as the disease progresses and the vertebral column deteriorates, the dogs become extremely fragile, and especially gentle handling is necessary when working with them or moving them between cages. Acute disk herniation can occur with even very minor trauma or inappropriate handling. Once skeletal disease has developed, exercise must be limited, and affected dogs must be protected from more rambunctious colony members. Decubital sores are a frequent consequence of the in- crease in time spent lying down. Housing affected dogs on shredded news- paper or elevated wire mesh provides both comfort and better sanitation. Appetite generally remains normal, although hand feeding or varying the diet might become necessary, especially in dogs with pronounced cor

110 DOGS: LABORATORY ANIMAL MANAGEMENT neat clouding, impaired hearing, or the rare decrease in cerebral sensorium. Some dogs have enlarged tongues; however, prehension of food is generally not a problem. The teeth of dogs that are fed a diet composed mainly of canned food require periodic scaling of tartar. MPS I-affected dogs are rarely maintained until they die naturally. By the age of 24-36 months, the symptoms of skeletal disease are generally so marked that euthanasia is indicated before debilitation becomes unaccept- able. Reproduction Most LSDs can impair fertility in dogs. MPS I- and VII-affected males have sired litters by artificial insemination. Males with fucosidosis show copulatory behavior before they become severely uncoordinated, but they are infertile because of epididymal lesions, which probably impair sperma- tozoan capacitation. Females with fucosidosis are fertile but are very poor mothers; their pups usually must be fostered or hand-reared. Pups with LSDs are generally produced by breeding heterozygous carriers that are clinically normal. Muscular Dystrophy Clinical Features A genetic disorder homologous to Duchenne's muscular dystrophy of humans a devastating, fatal disorder predominantly of boys- occurs in various breeds of dogs. The disorder in dogs, which is inherited as a simple sex-linked recessive gene with full penetrance, is known as canine X-linked muscular dystrophy, and dogs with the condition are called xmd dogs. The mutation has been found in golden retrievers and rottweilers, and a similar mutation is suspected to have occurred in Samoyeds, malamutes, and Irish terriers. The golden retriever is the best studied of the affected breeds, and the following discussion is based on data on this breed. Both Duchenne's muscular dystrophy and canine X-linked muscular dystrophy are caused by a defect in the production of dystrophin, a skeletal muscle cytoskeletal protein. The mutation in the dystrophin gene results in massive continuing skeletal muscle degeneration that occurs from birth on- ward. In dogs, progressive cardiac muscle degeneration begins in hemizy- gous males at the age of about 6 months. Carrier bitches appear clinically normal but have subtle lesions in their cardiac muscles. Because of the homology to Duchenne's muscular dystrophy, the xmd dog can serve as an animal model for studies leading to better understanding of the pathogen

SPECIAL CONSIDERATIONS 111 esis of Duchenne's muscular dystrophy, as well as for studies designed to assess therapeutic approaches (Valentine et al., 1992~. Clinical signs of obvious weakness, muscle wasting, and abnormal gait appear in xmd dogs at the age of about 8 weeks. After that time, clinical signs progress, and they are most severe at the age of about 6 months, at which time the dogs have a markedly stiff, shuffling gait. There is fre- quently a severely abnormal posture, with carpal overextension, tarsal over- flexion, and splaying of the limbs. The dogs are unable to open their jaws fully, their tongues are thickened and cannot be fully extended, and they frequently drool excessively. After the age of 6 months, the clinical disease appears to stabilize, and many dogs seem to gain strength as they age. However, there is still a progressive degeneration and fibrosis of cardiac muscle that results in the characteristic Duchenne-type cardiomyopathy. ~ . ~ Husbandry and Veterinary Care Dystrophic dogs do not require special caging. Shavings provide a soft, warm surface, but the shavings must be free of dust so that the dogs do not inhale particles and develop granulomatous pneumonia. Temperature and Older dystrophic dogs should be monitored carefully for signs of cardiac failure. Treatment for heart failure has been described (Fraser et al., 19911. Euthanasia should be considered when treatment fails to alleviate clinical signs (e.g., when the dog has diffi- culty breathing and when fluid accumulates in the abdomen). humidity must be carefully controlled. , ~ Dystrophic dogs require high-calorie food that is easy to prehend and swallow because of the weakness of their tongue, jaw, and esophageal muscles. Canned food mixed with moistened dry food seems to constitute an ad- equate diet, but careful monitoring of food intake and weight is necessary. Regurgitation of food is common because of the esophageal skeletal muscle dysfunction. Severely disabled dogs might not be able to use automatic watering devices and might have to be Liven water in bowls or buckets. _ _ _ '['heir water might need frequent chancing because of a buildup of saliva. Adequate exercise IS crucial during the period of rapid growth. Al- though dystrophic dogs might prefer to lie down, restricted exercise will result in more severe joint contractures. The presence of a slightly more active dystrophic cagemate is ideal, provided that competition for food does not impair food intake. The kennel must have a nonslippery surface to provide traction, and daily release for exercise is advised. These dogs should not be forced to exercise, however, because it might lead to in- creased muscle damage. Dystrophic dogs cannot groom themselves adequately. Regular brush- ing of their haircoat and clipping of overgrown toenails is required. To

2 DOGS: LABORATORY ANIMAL MANAGEMENT prevent skin irritation, the mouth and jaw should be kept free of the saliva and food that accumulate. Reproduction Many dystrophic dogs survive to breeding age, and breeding colonies can be established. Some affected males are able to breed naturally; others are hampered by their physical disability and require artificial insemination techniques. An xmd male that breeds naturally might need assistance to remain upright once he has "tied" with the female. Breeding dystrophic bitches, which are produced by mating dystrophic males to carrier bitches, is possible but not advised. Pregnant dystrophic bitches require constant monitoring, are likely to have respiratory and cardiac complications, will require cesarean section, and might not be able to care for their pups ade- quately. At whelping, a safe, warm environment and proper maternal care are essential for the survival of dystrophic pups. If dystrophic pups are stressed by cold, separation from the litter, or inability to compete with normal pups in a large litter, some of them will develop massive skeletal necrosis within the first few days of life. Once signs of severe weakness have developed in a pup, it is virtually impossible to save it. Severe diaphragmatic necrosis resulting in respiratory failure appears to be the cause of death. Dystrophic pups can be identified in the first week of life by their markedly increased serum concentrations of creatine kinase released from degenerating muscles. Dystrophic pups that survive the first week grow more slowly than their littermates. Euthanasia should be considered for pups that are too weak to nurse during the first week of life; tube feeding has not been successful in keeping such pups alive (B. A. Valentine, Department of Pathology, New York State College of Veterinary Medicine, Cornell University, Ithaca, N.Y., unpublished). Neurologic Disorders Clinical Features Dogs with hereditary or induced necrologic disorders are often used to study equivalent human disorders. Clinical signs in these dogs include abnormal gait, hyperactivity, nervousness, tremors, convulsions, visual im- pairment, blindness, deafness, quadriplegic, and tetraplegia. Obviously, these dogs commonly require extra care to ensure that they are as comfort- able as possible. Inherited canine necrologic diseases and their clinical signs have been reviewed by Cummings (1979) and Oliver and Lorenz

SPECIAL CONSIDERATIONS 113 (1-9931; the pattern of inheritance of specific diseases has been discussed by Willis (1989). Husbandry and Veterinary Care Food and water must be placed where a neurologically impaired dog can find and reach them easily, and, if the dog is blind, placement should be consistent. That might require using water bowls instead of automatic wa- terers or, for dogs with severe impairment, intravenous or subcutaneous administration of fluids. Food might have to be placed in flat dishes, soft- ened, or made into a gruel so that it can more easily be reached, masticated, and swallowed. Food and water intake should be monitored. Dogs should be weighed regularly to ensure that body weight is maintained. Nasogastric, ravage, pharyngotomy, or intragastric feeding might be required in some circumstances to provide adequate nutrition. Dogs with sensory deficits can experience dysesthesias and might re- spond by chewing the affected limb or body part or another, more acces- sible body part. Several strategies can be used to deal with such behavior. Dogs should be closely monitored to detect the beginning of self-directed behaviors. A dog can sometimes be distracted by housing it where it has more external visual and social stimulation. If a nonaggressive cagemate can be identified, social housing might be sufficiently distracting" provided that the cagemate does not harass the affected dog or prevent it from eating and drinking. Toys, such as rawhide bones, might also be useful. If ban- dages must be used, they should not be too tight and should be checked regularly. Elizabethan collars or muzzles can be used to limit access to the body. Light tranquilization, if it does not interfere with the experimental protocol, might be helpful. Dogs with sensory deficits might require extra or different bedding to prevent unintentional self-injury. The dogs' primary housing must be free of rough or sharp edges and projections. Dogs with motor deficits might have difficulty in positioning their bodies for urination and defecation. Some- times all that is necessary is to provide flooring with better traction (e.g., plastic-coated grids or rubber mats). If necessary, research or animal-care personnel should assist the dog to position itself. Catheterization or manu- ally expressing the bladder might be required to prevent urinary retention. Careful husbandry and nursing will avoid decubitus ulcers. In dogs with respiratory deficits, the normal ability to thermoregulate by panting has been compromised. For these animals, exertion must be avoided and comfortable temperatures maintained.

114 Reproduction DOGS: LABORATORY ANIMAL MANAGEMENT Dogs with some necrologic disorders can reproduce, even though they are severely impaired. Such dogs usually need assistance for mating or require artificial insemination. Bitches with marked sensory or motor defi- cits or ataxia should be closely attended at parturition and while nursing to protect the pups from accidental injury. If the necrologic deficits of the dam interfere with her ability to care for her offspring, hand rearing or foster rearing will be required. Ophthalmologic Disorders Clinical Features Dogs are affected by various ophthalmologic problems, either as inher- ent aspects of the research in which they are being used, as complications, or as acquired conditions unrelated to the research. Descriptions of canine eye diseases can be found in any standard text on veterinary ophthalmology (e.g., Gelatt, 1991; Helper, 1989~. In the research setting, ocular problems that require special management techniques are visual impairment, painful ocular conditions, untoward sequelae of interfering with the eye's external protective mechanisms, and combinations of these conditions. Blindness. Visual impairment in dogs usually cannot be measured pre- cisely. For purposes of this discussion, blindness is used, in a loosely defined manner, to refer to any condition in which visual impairment is sufficient to interfere with a dog's ability to perform visually guided tasks or to exhibit normal visually guided behavior. In general, dogs maintained in a familiar environment adapt well to visual deficits that are congenital, are gradual in onset, or have been present for an extended time (weeks to months). A dog that has adapted to its blindness, that is maintained in a familiar environment, and that is not subjected to stressful experiences will move about actively and engage in all normal canine behavior. Its adapta- tion, or compensation, might be so successful that a naive observer will not recognize that it is blind. Ocular pain. Painful ocular conditions fit broadly into three categories. External ocular pain is usually associated with corneal irritation and com- monly causes obvious signs, such as blinking, excessive tearing, and red- ness. Uveal pain is caused by intraocular inflammation, which might not be evident without careful examination of the eye; uveal pain is usually more painful than corneal irritation. Glaucomatous pain is often the most insidi- ous and most severe ocular pain. All these conditions are not only painful, but can threaten a dog's vision and the integrity of the affected eye.

SPECIAL CONSIDERATIONS 115 Conditions associated with failure of the eye's external protective mecha- nisms. Untoward sequelae can arise from any condition that interferes with the eye's external defense mechanisms. These mechanisms depend on such functions as corneal sensitivity, lid movement, and tear production. Any- thing that reduces corneal sensation, interferes with lid movement, or low- ers tear production can lead rapidly to painful ocular inflammation, impair- ment of vision, and loss of the affected eye. Common causes include anesthesia, radiation, surgical procedures, and drugs. Husbandry and Veterinary Care It is recommended that all experimental protocols involving dogs with ophthalmologic problems whether the problems are "natural" (i.e., genetic), acquired, or induced be reviewed by a veterinarian or a physician with training in ophthalmology (e.g., a veterinarian certified by the American College of Veterinary Ophthalmologists). Such protocols should include an adequate pro cram for monitoring the dogs' ophthalmologic problems and written procedures for dealing with ocular emergencies. Blindness. In spite of the ease with which dogs can adapt to blindness, they require special protection from a variety of environmental dangers, the more obvious of which are protruding objects, sharp edges, openings through which a dog might fall, and sources of electric or thermal injury. More insidious risks can arise because blind dogs lack the menace reflex, which normally protects the cornea from damage by causing the eyelids to blink in response to seen objects approaching the eye. Personnel responsible for the care and handling of blind dogs must be aware of these risks and keep them to a minimum and must watch for signs of acute or chronic corneal injury. Dogs that have adapted to their blindness can become decompensated in response to rapid changes in their environment or other stressful experi- ences, such as anesthesia (e.g., for diagnostic, surgical, or experimental procedures), illness, and alterations in their daily routine. A decompensated chronically blind dog might look as though it has suddenly become blind and might exhibit behaviors compatible with a general stress reaction- from stiff-limbed hesitancy in walking and an apparent fear of its surround- ings to anorexia or polydipsia, polyphagia, and polyuria. Similar signs can be observed in some dogs that have recently and rapidly lost their sight. Given time and a restricted, safe, and consistent environment, the blind dog will readapt and once again exhibit compensated normal behavior. Person- nel responsible for the care and handling of blind dogs must be aware that these dogs need consistent familiar surroundings and that they might react adversely to stressful experiences. When approaching a blind dog, animal technicians should talk to it so that the dog will be more likely to perceive the approach as friendly. .

116 DOGS: LABORATORY ANIMAL MANAGEMENT Ocular pain. Ocular pain can vary from moderate to excruciating. Dogs in ophthalmologic research colonies are often at risk of developing ocular pain, either as a direct result of a study or as an unpredictable occa- sional side effect. In some cases, particularly if the pain is chronic or develops gradually, it will not be readily apparent without special examina- tion procedures, especially if the observer is inexperienced. Personnel re- sponsible for the care and handling of dogs used in ophthalmologic research should suspect that ocular pain is present when there is periocular soiling or when there are behavioral changes, such as decreased activity, decreased appetite, increased yawning, and changes in vocalization patterns. Conditions associated with failure of the eye's external protective mecha- nisms. All protocols should be reviewed for potential adverse effects on external ocular defense mechanisms, and dogs subject to such risks should be monitored carefully for evidence of adverse effects. Reproduction Most dogs with ophthalmologic disorders can breed normally. Orthopedic Disorders Clinical Features Dogs serve as models for both canine and human orthopedic diseases. Spontaneous bone and joint diseases of dogs have been reviewed (Lipowitz et al., 1993; Newton and Nunamaker, 1985; Whittick, 1990; Young, 19791. Orthopedic diseases can also be induced in dogs. Husbandry and Veterinary Care When inducing an orthopedic disease in dogs, one must first evaluate the dogs to be certain that natural bone and joint diseases are absent. Radi- ography is used to diagnose hip dysplasia, osteochondrosis, osteoarthritis, elbow dysplasia, and patellar luxation. These are considered heritable dis- orders because offspring of affected parents often have them and they occur in siblings. Ideally, the surgical suite, the radiographic diagnostic facility, and an anesthesia recovery box lined with foam-rubber padding should be located near the primary housing facility. The floors of both the orthopedic re- search facility and the primary housing should be kept dry and have a nonslippery surface to provide good, steady footing.

SPECIAL CONSIDERATIONS 117 The amount of food consumed should be monitored because excess body weight will exacerbate orthopedic conditions. Limiting food con- sumption during the growth period has been shown to reduce signs of ortho- pedic disease in dogs that mature at greater than 30 lb (Kealy et al., 1992~. Dogs that refuse to eat because of pain might require a palatable high- energy food to maintain body weight. Human socialization is desirable to allow caregivers to detect abnormalities more readily and to facilitate han- dling and, when necessary, treatment. Mild exercise, such as walking, is beneficial to keep muscles limber, promote bone formation, and increase lubrication and nutrition of joints. However, excessive exercise aggravates pain and causes further bone or joint damage. Anti-inflammatory drugs, given with food, can be used to relieve pain. Glucocorticosteroids, although potent anti-inflammatory agents that relieve pain, can also accelerate disease progression and should be used only in advanced cases of joint disease. Warm packs can ease the pain of chronic osteoarthritis. Dogs affected with skeletal diseases should be kept warm and dry, although pain associated with a recent injury can be eased by applying crushed ice in a plastic bag to the affected region. Reproduction Dogs with joint and bone diseases can generally be bred, although it might be necessary to guide and hold a male affected with moderate or severe hip dysplasia. If the orthopedic problem is so severe that mating is not possible, artificial insemination can be used. Radiation Injury Clinical Features Radiation is commonly used in experimental protocols involving dogs. Total-body irradiation (TBI) is generally delivered by a cobalt-6O source or medical x-ray therapy machine. Doses of radiation up to 2 Gy can result in signs of illness related to mild gastrointestinal toxicity and decreased white- cell counts. At doses of 2-4 Gy, signs become progressively more severe. Doses greater than 4 Gy cause destruction of bone marrow, loss of circulat- ing blood cells, immunosuppression, increased tendency to bleed, and mod- erate to severe gastrointestinal toxicity. Bone-marrow transplantation can prevent severe clinical signs and death in dogs. The high radiation doses are similar to the doses that human transplantation patients receive. Several side effects occur in dogs that survive for long periods after TBI and bone-marrow rescue (Ladiges et al., 1989~: pancreatic fibrosis,

118 DOGS: LABORATORY ANIMAL MANAGEMENT malabsorption and malnutrition, radiation-induced cataracts, and malignan- cies. A consistent finding is graying of the hair. Radionuclides that are ingested, inhaled, or injected rarely produce signs of illness. However, knowledge of the chemical form and metabolism of the radionuclide is necessary to determine possible side effects. For ex- ample, inhaled particles of oxides of cesium-144 are relatively insoluble in the lungs and potentially remain there for some time. Signs of radiation pneumonitis might then be expected (Mauderly et al., 19804. Conversely, strontium-90 as a chloride is relatively soluble in the lungs. When inhaled, it is translocated to the bones, where it can cause prolonged thrombocytopenia and neutropenia (Gillett et al., 1987~. Types of radiation. Radiation emissions can be alpha particles, beta particles, gamma rays, and x rays. The distinctions between those emis- sions are important for providing care for laboratory animals. Alpha emissions from radionuclides, such as plutonium or americium, are generally high-energy emissions, but they travel very short distances in tissue. These radionuclides are rarely used in animals unless the study is specifically intended to assess metabolic or biologic effects of alpha emis- sions. No special precautions are needed for direct contact with animals contaminated with alpha-particle-emitting radionuclides because the radia- tion energy is absorbed within the animals' tissues. However, personnel should wear disposable clothing, shoe covers, gloves, eye protection, and respiratory protection to prevent inadvertent ingestion of, inhalation of, or wound contamination with alpha particles from contaminated feces, urine, bedding, cleaning water, or surfaces. Beta-emitting radionuclides, such as cesium-144 and strontium-90, pen- etrate farther into animal tissues than alpha particles but still only a short distance. The same precautions should be taken as are taken for alpha particles. Dogs can usually be handled without taking further precautions 10-12 days after administration of radionuclides. Gamma rays and x rays from internally deposited radionuclides pen- etrate tissues for considerable distances. These emissions can cause some radiation exposure of personnel, and it is important to know the potential exposure levels. These are generally low-energy kinds of radiation with short half-lives. Procedures for monitoring radiation must be in place to be certain that exposures of personnel are within accepted standards. The facility radiation-protection officer should participate in planning of ani- mal-care procedures. Disposal of radioactive wastes is regulated by both federal and state governments. It is important to have procedures in place for collecting, packaging, and labeling radioactive wastes before studies are initiated.

SPECIAL CONSIDERATIONS 119 Biohazards associated with radioactivity. Dogs exposed to external radiation sources do not pose a hazard to personnel once exposure is com- plete; the concern is for the effects on the health of the exposed animals. However, dogs that are administered radionuclides by ingestion, injection, or inhalation might present a continuing hazard to personnel because the radionuclide will be excreted in feces, in urine, and in some instances in exhaled air for some period after exposure. Standard operating procedures must be developed and followed for collecting and disposing of all contami- nated materials to protect animals and personnel. Animal health is of im- mediate concern only when large quantities of radionuclides are given. Husbandry and Veterinary Care Dogs exposed to external radiation can be housed in the usual manner (see Chapter 3~; however, it is critical that immunosuppressed dogs be pro- tected from other dogs that might harbor pathogens. Dogs given internally deposited radionuclides should be housed individually. To facilitate collec- tion of contaminated excrete and cage-cleaning water, the cages should be designed for collection of urine and feces and should be easy to clean. Dog rooms must have adequate ventilation, and ventilated air should not be recirculated. It might also be necessary to filter exhaust air. To prevent cross-contamination and simplify monitoring, it is recommended that dogs exposed to the same radionuclide be housed in the same room. Clinical observations and frequent peripheral-blood-cell counts are use- ful for monitoring dogs exposed to large doses of radiation. Treatment for reduced numbers of blood cells is supportive, and euthanasia should be considered if illness becomes too severe. Marrow "rescue" can prevent severe illness. Supportive care should consist of aggressive antibiotic and fluid therapy, and a semiliquid diet is necessary during the immediate post- irradiation period. Euthanasia should be considered in long-term survivors experiencing pancreatic fibrosis, malignancies, or pneumonitis. Reproduction Dogs that have received TBI are usually sterile. Lower doses of radia- tion have variable effects on reproduction. Gene Therapy Gene therapy can be used to correct inborn errors of metabolism, hemoglobinopathies, and blood factor A deficiencies; to insert genes into normal cells of the host (e.g., marrow stem cells) to increase their resistance to the toxic effects of chemotherapy; to introduce genes into cancer cells

120 DOGS: LABORATORY ANIMAL MANAGEMENT that will restore suppressor-gene function or neutralize the function of acti- vated oncogenes; and to induce tolerance to transplantation antigens by transferring genes that code for such antigens (Anderson, 1984~. The use of the dog as a preclinical, large, random-bred animal model has set the stage for clinical gene therapy. A number of target tissues for gene therapy have been used; this section will cover three of them. Hematopoietic Stem Cells In preparation for gene transfer, marrow is aspirated while the dog is under general anesthesia. The hair over the shoulder and hip joints is clipped. The skin is cleaned with povidone iodine, washed with 70 percent ethyl alcohol, and cleansed with sterile Ringer's solution. Under sterile conditions, a needle 20 cm long and 2.5 mm in internal diameter is inserted into the marrow cavity through the proximal intertubercular groove of the humerus or trochanteric fossa of the femur. The needle is then connected with polyvinyl tubing to a suction flask, and marrow is aspirated by placing a suction flask, which contains tissue-culture medium and preservative-free heparin, under negative pressure with a pump. The procedure can be com- pleted on all four limbs in approximately 20 minutes, during which 70-80 ml of a mixture of blood and bone marrow is collected. The marrow sus- pension is then passed through stainless-steel screens with 0.307- and 0.201- mm mesh diameters. A 1 ml sample is taken for marrow cell counts, and the remainder of the marrow is placed in plastic containers. The aspiration procedure is well tolerated without any sequelae. walking unimpaired after recovery from anesthesia. Nucleated marrow cells are then cocultivated with virus-producing packaging cells at a ratio of 2:1 for 24 hours in 850-ml roller bottles. The gene- containing vector is replication-defective. Retrovirus-producing packaging cells are seeded in roller bottles 48 hours before the addition of marrow and are cultured in vitro with established techniques. After cocultivation, mar- row cells are used to boost long-term cultures established 1 week earlier. The cultures are harvested after 6 days of incubation, and marrow cells are carefully removed without dislodging the virus-producing packaging cells, washed, resuspended in serum-free medium, and infused intravenously into the dog from which the marrow was taken. In preparation for the infusion, the dog is exposed to total-body irradia- tion to create room for the infused marrow to seed. Total-body irradiation is administered at doses of 4-10 Gy and is usually delivered at a rate of 7 cGy/minute from two opposing cobalt-60 sources. For that purpose, an unanesthetized dog is housed in a polyurethane cage that is midway be- tween the two cobalt-60 sources. The long axis of the cage is perpendicular to a line between the sources. After irradiation, the dog is returned to the Dogs are capable of

SPECIAL CONSIDERATIONS 121 animal-care facility for supportive care. Total-body irradiation can cause nausea, vomiting, and diarrhea. Its destruction of normal marrow leads to a disappearance of red cells, white cells, and platelets. The temporary ab- sence of those blood components produces a risk of anemia, infection, and bleeding that persists unless the dog receives a marrow graft and the graft begins to function. Dogs are monitored daily and receive parenteral fluids and electrolytes as required. Appropriate preoperative and postoperative antibiotics are routinely used to prevent and treat infections. Platelet and red-cell transfusions are given as needed. Marrow-graft function is moni- tored by evaluating daily blood counts. The success of gene transfer can be assessed by repeated aspiration of marrow under general anesthesia and examination of the samples for the appropriate marker gene with culture techniques, the polymerase chain re- action, or other appropriate methods (Stead et al., 1988~. Peripheral blood cells can be tested in a similar manner, as can lymph node lymphocytes and pulmonary macrophages (Stead et al., 1988~. Skin Keratinocytes Skin keratinocytes provide another good target for gene insertion. For some gene products, such as adenosine deaminase, gene transfer can take place in any replicating tissue. A 2 x 1.5-cm skin biopsy is obtained from the recipient under general anesthesia. Keratinocytes are derived from the biopsy material and cocultivated in vitro with replication-deficient retroviral vectors that contain the gene of interest. Keratinocytes are then cultured in a liquid-air interface culture, which gives rise to the various layers of skin in an in vitro system. After some time in culture, the skin grown in vitro is transplanted into a prepared bed on the flank of the dog under general anesthesia. The transplant site is treated with topical antibiotic powder, protected by nonadhering dressing, and inspected daily by the investigators. Generally, the skin grows in and is functional in 3-4 weeks. Punch biopsies of 2-3 mm allow assessment of gene transfer (Flowers et al., 1990~. Smooth Muscle Transplantation Because of their location, genetically modified vascular smooth muscle cells can be particularly useful for the treatment of some diseases (e.g., hemophilia). Studies have demonstrated that vascular smooth muscle cells are easily obtained, cultured, and genetically modified and replaced and provide a good target tissue for gene therapy that involves both secreted and nonsecreted proteins (Lim et al., 1991~. A segment of femoral artery or vein is surgically removed from a dog for preparation of smooth muscle cell cultures. The procedure of removing femoral artery and vein segments will

22 DOGS: LABORATORY ANIMAL MANAGEMENT not compromise the dog, because there is extensive collateral circulation in this region. With the dog under general anesthesia, as long a segment of vessel as possible (at least 2 cm) is isolated from the circulation with liga- tures. Any side branches in the two ends are permanently ligated before the vessel is removed. The smooth muscle cells are isolated, cultured, and infected with replication-defective amphotropic retroviruses that carry the genes of interest, in accordance with National Institutes of Health recombi- nant-DNA guidelines. The genetically modified smooth muscle cells are returned to the animal from which they were obtained. With the dog once again under general anesthesia, the transduced cells are seeded into the left and right carotid arteries and into the remaining femoral arteries (Lim et al., 1991~. REFERENCES Ackerman, N., R. Burk, A. W. Hahn, and H. M. Hayes, Jr. 1978. Patent ductus arteriosus in the dog: A retrospective study of radiographic, epidemiologic, and clinical findings. Am. J. Vet. Res. 39:1805-1810. Andersen, A. C., and M. E. Simpson. 1973. The Ovary and Reproductive Cycle of the Dog (Beagle). Los Altos, Calif.: Geron-X, Inc. 290 pp. Anderson, W. F. 1984. Prospects for human gene therapy. Science 226:401-409. Arbulu, A., S. N. Ganguly, and E. Robin. 1975. Tricuspid valvulectomy without prosthetic replacement: Five years later. Surg. Forum 26:244-245. AVMA (American Veterinary Medical Association). 1993. 1993 Report of the AVMA Panel on Euthanasia. J. Am. Vet. Med. Assoc. 202:229-249. Beierwaltes, W. H., and R. H. Nishiyama. 1968. Dog thyroiditis: Occurrence and similarity to Hashimoto's struma. Endocrinology 83:501 -508. Bell, S. C., S. D. Carter, and D. Bennet. 1991. Canine distemper viral antigens and antibodies in dogs with rheumatoid arthritis. Res. Vet. Sci. 50:64-68. Ben, L. K., J. Maselli, L. C. Keil, and I. A. Reid. 1984. Role of the renin-angiotensin system in the control of vasopressin and ACTH secretion during the development of renal hyper- tension in dogs. Hypertension 6:35-41. Bice, D. E., and B. A. Muggenburg. 1985. Effect of age on antibody responses after lung immunization. Am. Rev. Respir. Dis. 132:661-665. Blum, J. R., L. C. Cork, J. M. Morris, J. L. Olson, and J. A. Winkelstein. 1985. The clinical manifestations of a genetically determined deficiency of the third component of comple- ment in the dog. Clin. Immunol. Immunopathol. 34:304-315. Bonagura, J. D., ed. 1986. Section 4: Cardiovascular diseases. Pp. 319-424 in Current Veterinary Therapy. IX. Small Animal Practice, R. W. Kirk, ed. Philadelphia: W. B. Saunders. Bovee, K. C., M. P. Littman, F. Saleh, R. Beeuwkes, W. Mann, P. Koster, and L. B. Kinter. 1986. Essential hereditary hypertension in dogs: A new animal model. J. Hypertens. 4(Suppl. 5):S172-S173. Brooks, D. P., and T. A. Fredrickson. 1992. Use of ameroid constrictors in the development of repin-dependent hypertension in dogs. Lab. Anim. Sci. 42:67-69. Brooks, D. P., T. A. Fredrickson, P. F. Koster, and R. R. Ruffolo, Jr. 1991. Effect of the dopamine p-hydroxylase inhibitor, SK&F 102698, on blood pressure in the 1-kidney, 1- clip hypertensive dog. Pharmacology 43:90-95.

SPECIAL CONSIDERATIONS 123 Brown-Sequard, E. 1856. Recherches experimentales sur la physiologic et la pathologic des capsules surrenales. Arch. Gen. Med. (Ser. 5)8(II):385-401. Buchanan, J. W. 1992. Causes and prevalence of cardiovascular disease. Pp. 647-654 in Current Veterinary Therapy XI, R. W. Kirk and J. D. Bonagura, eds. Philadelphia: W. B. Saunders. Bull, R. W., R. Schirmer, and A. J. Bowdler. 1971. Autoimmune hemolytic disease in the dog. J. Am. Vet. Med. Assoc. 159:880-884. Campbell, K. L. 1991. Immunoglobulin A deficiency in the dog: A retrospective study of 155 cases (1983-1990). Canine Pract. 16(4):7-11. Capen, C. C., and S. L. Martin. 1989. The thyroid gland. Pp. 58-91 in Veterinary Endocrinol- ogy and Reproduction, 4th ea., L. E. McDonald and M. H. Pineda, eds. Philadelphia: Lea & Febiger. Carlson, G. P. 1989. Fluid, electrolyte, and acid-base balance. Pp. 543-575 in Clinical Biochemistry of Domestic Animals, 4th ea., J. J. Kaneko, ed. San Diego: Academic Press. Carter, S. D., S. C. Bell, A. S. M. Bari, and D. Bennett. 1989. Immune complexes and rheumatoid factors in canine arthritides. Ann. Rheum. Dis. 48:986-991. Chester, D. K. 1987. The thyroid gland and thyroid diseases. Pp. 83-120 in Small Animal Endocrinology, F. H. Drazner, ed. New York: Churchill Livingstone. Chick, T. W., S. E. Goldblum, N. D. Smith, C. Butler, B. J. Skipper, J. A. Winkelstein, L. C. Cork, and W. P. Reed. 1984. Pneumococcal-induced pulmonary leukostasis and hemo- dynamic changes: Role of complement and granulocytes. J. Lab. Clin. Med. 103:180- 192. Cork, L. C., J. M. Morris, J. L. Olson, S. Krakowka, A. J. Swift, and J. A. Winkelstein. 1991. Membranoproliferative glomerulonephritis in dogs with a genetically determined defi- ciency of the third component of complement. Clin. Immunol. Immunopathol. 60:455- 470. Cummings, J. F., ed. 1979. Part XIII: Nervous system. Pp. 107-178 in Spontaneous Animal Models of Human Disease, vol. II, E. J. Andrews, B. C. Ward, and N. H. Altman, eds. New York: Academic Press. DePaolo, L. V., and E. J. Masoro. 1989. Endocrine hormones in laboratory animals. Pp. 279- 308 in The Clinical Chemistry of Laboratory Animals, W. F. Loeb and F. W. Quimby, eds. New York: Pergamon Press. De Reeder, E. G., N. Girard, R. E. Poelmann, J. C. Van Munsteren, D. F. Patterson, and A. C. Gittenberger-de Groot. 1988. Hyaluronic acid accumulation and endothelial cell detach- ment in intimal thickening of the vessel wall: The normal and genetically defective ductus arteriosus. Am. J. Pathol. 132:574-585. De Rick, A., F. M. Belpaire, M. G. Bogaert, and D. Mattheeuws. 1978. Plasma concentrations of digoxin and digitoxin during digitalization of healthy dogs and dogs with cardiac failure. Am. J. Vet. Res. 39:811 -815. DiBartola, S. P., M. J. Tarr, D. M. Webb, and U. Giger. 1990. Familial renal amyloidosis in Chinese Shar Pei dogs. J. Am. Vet. Med. Assoc. 197:483-487. Dodds, W. J. 1983. Immune-mediated diseases of the blood. Adv. Vet. Sci. Comp. Med. 27: 163-196. Dodds, W. J. 1988. Third international registry of animal models of thrombosis and hemor- rhagic diseases. ILAR News 30:R1-R32. Dodds, W. J. 1989. Hemostasis. Pp. 274-315 in Clinical Biochemistry of Domestic Animals, 4th ea., J. J. Kaneko, ed. San Diego: Academic Press. Dodds, W. J. 1992. Bleeding disorders. Pp. 765-777 in Handbook of Small Animal Practice, 2d ea., R. V. Morgan, ed. New York: Churchill Livingstone. Dougherty, S. H. 1986. Implant infections. Pp. 276-289 in Handbook of Biomaterials Evalu- ation, A. F. von Recum, ed. New York: Macmillan.

24 DOGS: LABORATORY ANIMAL MANAGEMENT Drazner, F. H. 1987a. The adrenal cortex. Pp. 201-277 in Small Animal Endocrinology, F. H. Drazner, ed. New York: Churchill Livingstone. Drazner, F. H., ed. 1987b. Small Animal Endocrinology. New York: Churchill Livingston. 508 pp. Eigenmann, J. E. 1985. Acromegaly. Model no. 311 in A Handbook: Animal Models of Human Disease, fascicle 14, C. C. Capen, T. C. Jones, and G. Migaki, eds. Washington, D.C.: Registry of Comparative Pathology, Armed Forces Institute of Pathology. Eigenmann, J. E. 1989. Pituitary-hypothalamic diseases. Pp. 1579-1609 in Textbook of Veterinary Internal Medicine, vol. 2, 3rd ea., S. J. Ettinger, ed. Philadelphia: W. B. Saunders. Ettinger, S. J., ed. 1989. Textbook of Veterinary Internal Medicine, vol. 2, 3rd. ed. Philadel- phia: W.B. Saunders. 1,237 pp. Eyster, G. E. 1992. Congenital diseases. Pp. 63-69 in Handbook of Small Animal Practice, 2d ea., R. V. Morgan, ed. New York: Churchill Livingstone. Feldman, E. C. 1989. Adrenal gland disease. Pp. 1721-1774 in Textbook of Veterinary Internal Medicine, vol. 2, 3rd ea., S. J. Ettinger, ed. Philadelphia: W. B. Saunders. Feldman, E. C., and R. W. Nelson. 1987. Canine and Feline Endocrinology and Reproduc tion. Philadelphia: W. B. Saunders. 564 pp. Felsburg, P. J., L. T. Glickman, and P. F. Jezyk. 1985. Selective IgA deficiency in the dog. Clin. Immunol. Immunopathol. 36:297-305. Ferguson, D. C. 1986. Thyroid hormone replacement therapy. Pp. 1018- 1019 in Current Veterinary Therapy IX, R. W. Kirk, ed. Philadelphia: W. B. Saunders. Ferrario, C. M., C. Blumle, G. R. Nadzam, and J. W. McCubbin. 1971. An externally adjustable renal artery clamp. J. Appl. Physiol. 31:635-637. Fischer, C. A. 1989. Geriatric ophthalmology. Vet. Clinics N. Am. 19(1):103-123. Fixler, D. E., J. P. Archie, D. J. Ullyot, G. D. Buckberg, and J. I. E. Hoffman. 1973. Effects of acute right ventricular systolic hypertension on regional myocardial blood flow in anesthetized dogs. Am. Heart J. 85:491 -500. Flowers, M. E. D., M. A. R. Stockschlaeder, F. G. Schuening, D. Niederwieser, R. Hackman, A. D. Miller, and R. Storb. 1990. Long-term transplantation of canine keratinocytes made resistant to G418 through retrovirus-mediated gene transfer. Proc. Natl. Acad. Sci. USA 87:2349-2353. Fraser, C. M., J. A. Bergeron, A. Mays, and S. E. Aiello, eds. 1991. Heart disease. Pp. 40-52 in The Merck Veterinary Manual: A Handbook of Diagnosis, Therapy, and Disease Prevention for the Veterinarian, 7th ed. Rahway, N.J.: Merck & Co. Gardner, T. J., and D. L. Johnson. 1988. Cardiovascular system. Pp. 74-113 in Experimental Surgery and Physiology: Induced Animal Models of Human Disease, M. M. Swindle and R. J. Adams, eds. Baltimore: Williams & Wilkins. Gelatt, K. N., ed. 1991. Veterinary Ophthalmology, 2d ed. Philadelphia: Lea & Febiger. 765 pp. Gillett, N. A., B. A. Muggenburg, B. B. Boecker, F. F. Hahn, F. A. Seiler, A. H. Rebar, R. K. Jones, and R. O. McClellan. 1987. Single inhalation exposure to 9°SrC12 in the beagle dog: Hematological effects. Radiat. Res. 110:267-288. Gittenberger-de Groot, M. D., J. L. M. Strengers, M. Mentink, R. E. Poelmann, and D. F. Patterson. 1985. Histologic studies on normal and persistent ductus arteriosus in the dog. J. Am. Coll. Cardiol. 6:394-404. Goldston, R. T., ed. 1989. Geriatrics and gerontology. Vet. Clin. N. Am. 19(1):1-202. Gosselin, S. J., C. C. Capen, S. L. Martin, and S. Krakowka. 1982. Autoimmune lymphocytic thyroiditis in dogs. Vet. Immunol. Immunopathol. 3:185-201. Grindem, C. B., and K. H. Johnson. 1983. Systemic lupus erythematosus: Literature review and report of 42 new canine cases. J. Am. Anim. Hosp. Assoc. 19:489-503. Guyton, A. C. 1991. Dominant role of the kidneys in long-term regulation of arterial pressure

SPECIAL CONSIDERATIONS 125 and in hypertension: The integrated system for pressure control. Pp. 205-220 in Text- book of Medical Physiology, 8th ed. Philadelphia: W. B. Saunders. Haley, P. J., F. F. Hahn, B. A. Muggenburg, and W. C. Griffith. 1989. Thyroid neoplasms in a colony of beagle dogs. Vet. Pathol. 26:438-441. Hall, R. L., and U. Giger. 1992. Disorders of red blood cells. Pp. 715-733 in Handbook of Small Animal Practice, 2d ea., R. V. Morgan, ed. New York: Churchill Livingstone. Harvey, J. W. 1989. Erythrocyte metabolism. Pp. 186-234 in Clinical Biochemistry of Domestic Animals, 4th ea., J. J. Kaneko, ed. San Diego: Academic Press. Haskins, M. E., R. J. Desnick, N. DiFerrante, P. F. Jezyk, and D. F. Patterson. 1984. 13- glucuronidase deficiency in a dog: A model of human mucopolysaccharidosis VII. Pediatr. Res. 18:980-984. Healy, P. J., B. R. H. Farrow, F. W. Nicholas, K. Hedberg, and R. Ratcliffe. 1984. Canine fucosidosis: A biochemical and genetic investigation. Res. Vet. Sci. 36:354-359. Hegreberg, G. A., G. A. Padgett, J. R. Gorham, and J. B. Henson. 1969. A connective tissue disease of dogs and mink resembling the Ehlers-Danlos syndrome of man. II. Mode of inheritance. J. Hered. 60:249-254. Hegreberg, G. A., G. A. Padgett, R. L. Ott, and J. B. Henson. 1970. A heritable connective tissue disease of dogs and mink resembling the Ehlers-Danlos syndrome of man. I. Skin tensile strength properties. J. Invest. Dermatol. 54:377-380. Helper, L. C. 1989. Magrane's Canine Ophthalmology, 4th ed. Philadelphia: Lea & Febiger. 297 pp. Hsu, W. H., and M. H. Crump. 1989. The adrenal gland. Pp. 202-230 in Veterinary Endocri- nology and Reproduction, 4th ea., L. E. McDonald and M. H. Pineda, eds. Philadelphia: Lea & Febiger. Jarvinen, A.-K. 1981. Urogenital tract infection in the bitch. Vet. Res. Commun. 4:253-269. Jezyk, P. F., P. J. Felsburg, M. E. Haskins, and D. F. Patterson. 1989. X-linked severe combined immunodeficiency in the dog. Clin. Immunol. Immunopathol. 52:173-189. Johnson, J. P., R. H. McLean, L. C. Cork, and J. A. Winkelstein. 1986. Animal model: Genetic analysis of an inherited deficiency of the third component of complement in Brittany spaniel dogs. Am. J. Med. Genet. 25:557-562. Kaneko, J. J. 1987. Critical review. Animal models of inherited hematologic disease. Clin. Chim. Acta 165: 1 - 19. Kaneko, J. J. 1989. Carbohydrate metabolism and its diseases. Pp. 44-85 in Clinical Bio- chemistry of Domestic Animals, 4th ea., J. J. Kaneko, ed. San Diego: Academic Press. Kaplan, A. V., and F. W. Quimby. 1983. A radiolabeled staphylococcal protein A assay for detection of anti-erythrocyte IgG in warm agglutinin autoimmune hemolytic anemia of dogs and man. Vet. Immunol. Immunopathol. 4:307-317. Kaswan, R. L., C. L. Martin, and D. L. Dawe. 1985. Keratoconjunctivitis sicca: Immunologi- cal evaluation of 62 canine cases. Am. J. Vet. Res. 46: 376-383. 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 hipdysplasia in growing dogs. J. Am. Vet. Med. Assoc. 201:857-863. Kesel, M. L., and D. H. Neil. 1990. Restraint and handling of animals. Pp. 1-30 in Clinical Textbook for Veterinary Technicians, 2d ea., D. M. McCurnin, ed. Philadelphia: W. B. Saunders. Kirk, R. W., and S. I. Bistner. 1985. Metabolic emergencies. Pp. 138-149 in Handbook of Veterinary Procedures and Emergency Treatment, 4th ed. Philadelphia: W. B. Saunders. Klag, A. R., U. Giger, and F. S. Shofer. 1993. Idiopathic immune-mediated hemolytic anemia in dogs: 42 cases (1986-1990). J. Am. Vet. Med. Assoc. 202:783-788. Knight, D. H., D. F. Patterson, and J. Melbin. 1973. Constriction of the fetal ductus arteriosus induced by oxygen, acetylcholine, and norepinephrine in normal dogs and those geneti- cally predisposed to persistent patency. Circulation 47:127-132.

26 DOGS: LABORATORY ANIMAL MANAGEMENT Knoll, J. S. 1992. Disorders of white blood cells. Pp. 735-749 in Handbook of Small Animal Practice, 2d ea., R. V. Morgan, ed. New York: Churchill Livingstone. Krakowka, S., L. C. Cork, J. A. Winklestein, and M. K. Axthelm. 1987. Establishment of central nervous system infection by canine distemper virus: Breach of the blood-brain barrier and facilitation by antiviral antibody. Vet. Immunol. Immunopathol. 17:471-482. Kramer, J. W. 1981. Inherited early-onset, insulin-requiring diabetes mellitus in keeshond dogs. Am. J. Pathol. 105: 194- 196. Ladiges, W. C., H. J. Deeg, J. A. Aprile, R. F. Raff, F. Schuening, and R. Storb. 1988. Differentiation and function of lymphohemopoietic cells in the dog. Pp. 307-335 in Differentiation Antigens in Lymphohemopoietic Tissues, M. Miyasaka and Z. Trnka, eds. New York: Marcel Dekker. Ladiges, W. C., R. Storb, T. Graham, and E. D. Thomas. 1989. Experimental techniques used to study the immune system of dogs and other large animals. Pp. 103-133 in Methods of Animal Experimentation, vol. VII, part C, W. I. Gay and J. E. Heavner, eds. New York: Academic Press. Ladiges, W. C., R. Storb, and E. D. Thomas. 1990. Canine models of bone marrow transplan- tation. Lab. Anim. Sci. 40: 11 - 15. Lage, A. L., N. A. Gillett, R. F. Gerlach, and E. N. Allred. 1989. The prevalence and distribution of proliferative and metaplastic changes in normal appearing canine bladders. J. Urol. 141:993-997. Lange, J., B. Brockway, and S. Azar. 1991. Telemetric monitoring of laboratory animals: An advanced technique that has come of age. Lab Anim. 20(7):28-33. Lim, C. S., G. D. Chapman, R. S. Gammon, J. B. Muhlestein, R. P. Bauman, R. S. Stack, and J. L. Swain. 1991. Direct in vivo gene transfer into the coronary and peripheral vascula- tures of the intact dog. Circulation 83 :2007-2011. Lipowitz, A. J., D. D. Caywood, C. D. Newton, and M. E. Finch. 1993. Small Animal Orthopedics Illustrated: Surgical Approaches and Procedures. St. Louis: Mosby. 336 pp. Lowseth, L. A., N. A. Gillett, R. F. Gerlach, and B. A. Muggenburg. 1990a. The effects of aging on hematology and serum chemistry values in the beagle dog. Vet. Clin. Pathol. 19(1):13-19. Lowseth, L. A., R. F. Gerlach, N. A. Gillett, and B. A. Muggenburg. 1990b. Age-related changes in the prostate and testes of the beagle dog. Vet. Pathol. 27:347-353. Lund, J. E., G. A. Padgett, and R. L. Ott. 1967. Cyclic neutropenia in grey collie dogs. Blood 29:452-461. MacVean, D. W., A. W. Monlux, P. S. Anderson, Jr., S. L. Silberg, and J. F. Rozel. 1978. Frequency of canine and feline tumors in a defined population. Vet. Pathol. 15:700-715. Maggio-Price, L., C. L. Emerson, T. R. Hinds, F. F. Vincenzi, and W. R. Hammond. 1988. Hereditary nonspherocytic hemolytic anemia in beagles. Am. J. Vet. Res. 49:1020-1025. Mann, W. A., M. S. Landi, E. Homer, P. Woodward, S. Campbell, and L. B. Kinter. 1987. A simple procedure for direct blood pressure measurements in conscious dogs. Lab. Anim. Sci. 37:105-108. Mauderly, J. L., and F. F. Hahn. 1982. The effects of age on lung function and structure of adult animals. Adv. Vet. Sci. Comp. Med. 26:35-77. Mauderly, J. L., B. A. Muggenburg, F. F. Hahn, and B. B. Boecker. 1980. The effects of inhaled 144Ce on cardiopulmonary function and histopathology of the dog. Radiat. Res. 84:307-324. McCarthy, C. R., and J. G. Miller. 1990. OPRR Reports, May 21, 1990. Available from Office for Protection from Research Risks (OPRR), Building 31, Room 5B59, National Institutes of Health, Bethesda, MD 20892. McDonald, L. E., and M. H. Pineda, eds. 1989. Veterinary Endocrinology and Reproduction, 4th ed. 571 pp.

SPECIAL CONSIDERATIONS 127 Meuten, D. J., C. C. Capen, G. J. Kociba, and B. J. Cooper. 1982. Hypercalcemia of malig- nancy. Hypercalcemia associated with an adenocarcinoma of the apocrine glands of the anal sac. Am. J. Pathol. 108:366-370. Meuten, D. J., C. C. Capen, and G. J. Kociba. 1986. Hypercalcemia of malignancy. Supple- mental update, 1986: Model no. 143 in A Handbook: Animal Models of Human Disease, fascicle 15, C. C. Capen, T. C. Jones, and G. Migaki, eds. Washington, D.C.: Registry of Comparative Pathology, Armed Forces Institute of Pathology. Mill, A. B., and K. L. Campbell. 1992. Concurrent hypothyroidism, IgM deficiency, impaired T-cell mitogen response, and multifocal cutaneous squamous papillomas in a dog. Ca- nine Pract. 17(2): 15-21. Milne, K. L., and H. M. Hayes, Jr. 1981. Epidemiologic features of canine hypothyroidism. Cornell Vet. 73:3-14. Minor, R. R., J. A. M. Wootton, D. J. Prockop, and D. F. Patterson. 1987. Genetic diseases of connective tissues in animals. Curr. Probl. Dermatol. 17:199-215. Mizejewski, G. J., J. Baron, and G. Poissant. 1971. Immunologic investigations of naturally occurring canine thyroiditis. J. Immunol. 107: 1152- 1160. Monier, J. C., C. Fournel, M. Lapras, M. Dardenne, T. Randle, and C.M. Fontaine. 1988. Systemic lupus erythematosus in a colony of dogs. Am. J. Vet. Res. 49:46-51. Mordes, J. P., and A. A. Rossini. 1985. Animal models of diabetes mellitus. Pp. 110-137 in Joslin's Diabetes Mellitus, 12th ea., A. Marble, L. P. Krall, R. F. Bradley, A. R. Christlieb, and J. S. Soeldner, eds. Philadelphia: Lea & Febiger. Morgan, R. V, ed. 1992. Handbook of Small Animal Practice, 2d ed. New York: Churchill Livingstone. 1,513 pp. Moroff, S. D., A. I. Hurvitz, M. E. Peterson, L. Saunders, and K. E. Noone. 1986. IgA deficiency in Shar-Pei dogs. Vet. Immunol. Immunopathol. 13:181-188. Nakano, K., M. M. Swindle, F. G. Spinale, K. Ishihara, S. Kanazawa, A. Smith, R. W. W. Biederman, L. Clamp, Y. Hamada, M. R. Zile, and B. A. Carabello. 1991. Depressed contractile function due to canine mitral regurgitation improves after correction of the volume overload. J. Clin. Invest. 87:2077-2086. Nelson, A. A., and G. Woodard. 1949. Severe adrenal cortical atrophy (cytotoxic) and hepatic damage produced in dogs by feeding 2,2-bis(parachlorophenyl)-1,1-dichloroethane (000 or TDE). Arch. Pathol. 48:387-394. Nelson, R. W. 1989. Disorders of the endocrine pancreas. Pp. 1676-1720 in Textbook of Veterinary Internal Medicine, vol. 2, 3rd ea., S. J. Ettinger, ed. Philadelphia: W. B. Saunders. Newton, C. D., and D. M. Nunamaker. 1985. Textbook of Small Animal Orthopaedics. Philadelphia: J. B. Lippincott. 1,140 pp. Nichols, R., and M. E. Peterson. 1992. Hypoadenocorticism. Pp. 531-534 in Handbook of Small Animal Practice, 2d ea., R. V. Morgan, ed. New York: Churchill Livingstone. NRC (National Research Council), Institute of Laboratory Animal Resources, Committee on Care and Use of Laboratory Animals. 1985. 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), Institute of Laboratory Animal Resources, Committee on Immunologically Compromised Rodents. 1989. Introduction. Pp. 1-35 in Immunodefi- cient Rodents: A Guide to Their Immunobiology, Husbandry, and Use. Washington, D.C.: National Academy Press. Ogilive, G. K., W. M. Haschek, S. J. Withrow, R. C. Richardson, H. J. Harvey, R. A. Henderson, J. D. Fowler, A. M. Norris, J. Tomlinson, D. McCaw, J. S. Klausner, R. W. Reschke, and B. C. McKiernan. 1989. Classification of primary lung tumors in dogs: 210 cases (1975-1985). J. Am. Vet. Med. Assoc. 195:106-108. !

28 DOGS: LABORATORY ANIMAL MANAGEMENT O'Kane, H. O., A. S. Geha, R. E. Kleiger, T. Abe, M. T. Salaymeh, and A. B. Malik. 1973. Stable left ventricular hypertrophy in the dog. Experimental production, time course, and natural history. J. Thorac. Cardiovasc. Surg. 65:264-271. Oliver, J. E. Jr., and M. D. Lorenz. 1993. Appendix. Pp. 374-393 in Handbook of Veterinary Neurology, 2d ed. Philadelphia: W. B. Saunders. O'Neil, K. M., H. D. Ochs, S. R. Heller, L. C. Cork, J. M. Morris, and J. A. Winkelstein. 1988. Role of C3 in Immoral immunity. Defective antibody production in C3-deficient dogs. J. Immunol. 140:1939-1945. Patterson, D. F. 1968. Epidemiologic and genetic studies of congenital heart disease in the dog. Circ. Res. 23:171-202. Patterson, D. F. 1984. Two hereditary forms of ventricular outflow obstruction in the dog: Pulmonary valve dysplasia and discrete subaortic stenosis. Pp. 43-63 in Congenital Heart Disease: Causes and Processes, J. J. Nora and A. Takao, eds. Mt. Kisco, N.Y.: Future Publishing Co. Patterson, D. F., R. L. Pyle, J. W. Buchanan, E. Trautvetter, and D. A. Abt. 1971. Hereditary patent ductus arteriosus and its sequelae in the dog. Circ. Res. 29:1-13. Patterson, D. F., R. L. Pyle, L. Van Mierop, J. Melbin, and M. Olson. 1974. Hereditary defects of the conotruncal septum in keeshond dogs: Pathologic and genetic studies. Am. J. Cardiol. 34:187-205. Patterson, D. F., M. E. Haskins, and W. R. Schnarr. 1981. Hereditary dysplasia of the pulmonary valve in beagle dogs: Pathologic and genetic studies. Am. J. Cardiol. 47:631- 641. Patterson, D. F., M. E. Haskins, and P. F. Jezyk. 1982. Models of human genetic disease in domestic animals. Adv. Hum. Genet. 12:263-339. Patterson, D. F., T. Pexieder, W. R. Schnarr, T. Navratil, and R. Alaili. 1993. A single major- gene defect underlying cardiac conotruncal malformations interferes with myocardial growth during embryonic development: Studies in the CTD line of keeshond dogs. Am. J. Hum. Genet. 52:388-397. Petersen, J. C., R. R. Linartz, R. L. Hamlin, and R. E. Stoll. 1988. Noninvasive measurement of systemic arterial blood pressure in the conscious beagle dog. Fundam. Appl. Toxicol. 10:89-97. Peterson, M. E., and D. C. Ferguson. 1989. Thyroid disease. Pp. 1632-1675 in Textbook of Veterinary Internal Medicine, vol. 2, 3rd ea., S. J. Ettinger, ed. Philadelphia: W. B. Saunders. PHS (Public Health Service). 1986. Public Health Service Policy on Humane Care and Use of Laboratory Animals. Washington, D.C.: U.S. Department of Health and Human Ser- vices. 28 pp. Available from the Office for Protection from Research Risks, Building 31, Room 4B09, NIH, Bethesda, MD 20892. Plechner, A. J. 1979. IgM deficiency in 2 doberman pinschers. Mod. Vet. Pract. 60:150. Pyle, R. L., D. F. Patterson, and S. Chacko. 1976. The genetics and pathology of discrete subaortic stenosis in the Newfoundland dog. Am. Heart J. 92:324-334. Quimby, F. W. 1981. Canine systemic lupus erythematosus. Pp. 175-184 in Immunologic Defects in Laboratory Animals, vol. 2, M. E. Gershwin and B. Merchant, eds. New York: Plenum Press. Quimby, F. W., and R. S. Schwartz. 1978. The etiopathogenesis of systemic lupus erythematosus. Pathobiol. Annul 8:35-59. Quimby, F. W., C. Jensen, D. Nawrocki, and P. Scollin. 1978. Selected autoimmune diseases in the dog. Vet. Clin. N. Am. 8(4):665-682. Quimby, F. W., R. S. Schwartz, T. Poskitt, and R. M. Lewis. 1979. A disorder of dogs resembling Sjogren's syndrome. Clin. Imm-unol. Immunopathol. 12:471-476. Quimby, F. W., C. Smith, M. Brushwein, and R.W. Lewis. 1980. Efficacy of immunoserodiagnostic

SPECIAL CONSIDERATIONS 129 procedures in the recognition of canine immunologic diseases. Am. J. Vet. Res. 41:1662- 1666. Rajatanavin, R., S.-L. Fang, S. Pino, P. Laurberg, L. Braverman, M. Smith, and L. P. Bullock. 1989. Thyroid hormone antibodies and Hashimoto's thyroiditis in mongrel dogs. Endo- crinology 124:2535-2540. Renshaw, H. W., and W. C. Davis. 1979. Canine granulocytopathy syndrome. An inherited disorder of leukocyte function. Am. J. Pathol. 95:731-744. Rivas, A. L., L. Tintle, E. S. Kimball, J. Scarlett, and F. W. Quimby. 1992. A canine febrile disorder associated with elevated interleukin-6. Clin. Immunol. Immunopathol. 64:36-45. Ross, L. A. 1989. Hypertensive disease. Pp. 2047-2056 in Textbook of Veterinary Internal Medicine, vol. 2, 3rd ea., S. J. Ettinger, ed. Philadelphia: W. B. Saunders. Roth, J. A., L. G. Lomax, N. Altszuler, J. Hampshire, M. I. Kaeberle, M. Shelton, D. D. Draper, and A. E. Ledet. 1980. Thymic abnormalities and growth hormone deficiency in dogs. Am. J. Vet. Res. 41:1256-1262. Schrader, L. A. 1988. Hypoadrenocorticism. Pp. 543-546 in Handbook of Small Animal Practice, 2d ea., R. V. Morgan, eds. New York: Churchill Livingstone. Schwartz, R. S., F. W. Quimby, and J. Andre-Schwartz. 1978. Canine systemic lupus erythematosus: Phenotypic expression of autoimmunity in a closed colony. Pp. 287-294 in Genetic Control of Autoimmune Disease, N. R. Rose, P. Bigazzi, and N. Warner, eds. New York: Elsevier-North Holland. Shull, R. M., R. J. Munger, E. Spellacy, C. W. Hall, G. Constantopoulos, E. F. Neufeld. 1982. Canine a-~-iduronidase deficiency: A model of mucopolysaccharidosis I. Am. J. Pathol. 109:244-248. Smith, G. S., and J. H. Lumsden. 1983. Review of neutrophil adherence, chemotaxis, phago- cytosis and killing. Adv. Vet. Immunol. 1982 12:177-236. Stead, R. B., W. W. Kwok, R. Storb, and A. D. Miller. 1988. Canine model for gene therapy: Inefficient gene expression in dogs reconstituted with autologous marrow infected with retroviral vectors. Blood 71:742-747. Swindle, M. M., F. G. Spinale, A. C. Smith, R. E. Schumann, C. T. Green, K. Nakano, S. Kanasawa, K. Ishihara, M. R. Zile, and B. A. Carabello. 1991. Anesthetic and postop- erative protocols for a canine model of reversible left ventricular volume overload. J. Invest. Surg. 4:339-346. Taylor, G. N., L. Shabestari, J. Williams, C. W. Mays, W. Angus, and S. McFarland. 1976. Mammary neoplasia in a closed beagle colony. Cancer Res. 36:2740-2743. Terman, D. S., D. Moore, J. Collins, B. Johnston, D. Person, J. Templeton, R. Poser, and F. Quimby. 1979. Detection of immune complexes in sera of dogs with rheumatic and neoplastic diseases by 125I-Clq binding test. ~ Cnmn P~th~] S?O 991_~ ~^^r . ~^A~. ~ . ~ ~ ~ ~ I . Thacker, E. L., K. R. Refsal, and R. W. Bull. 1992. Prevalence of autoantibodies to thyroglo- bulin, thyroxine, or triiodothyronine and relationship of autoantibodies and serum con- centrations of iodothyronines in dogs. Am. J. Vet. Res. 53:449-453. Tholen, M. A., and R. F. Hoyt, Jr. 1983. Oral pathology. Pp. 39-67 in Concepts in Veterinary Dentistry. Edwardsville, Kansas: Veterinary Medicine Publishing Co. Valentine, B. A., N. J. Winand, D. Pradhan, N. S. Moise, A. de Lahunta, J. N. Kornegay, and B. J. Cooper. 1992. Canine X-linked muscular dystrophy as an animal model of Duchenne muscular dystrophy: A review. Am. J. Med. Genetics 42:352-356. Van Mierop, L. H. S., D. F. Patterson, and W. R. Schnarr. 1977. Hereditary conotruncal septal defects in keeshond dogs: Embryologic studies. Am. J. Cardiol. 40:936-950. Vlahakes, G. J., K. Turley, and J. I. E. Hoffman. 1981. The pathophysiology of failure in acute right ventricular hypertension: Hemodynamic and biochemical correlations. Cir- culation 63:87-95. Waye, J. W. 1960. Idiopathic thrombocytopenic purpura in a dog. Can. Vet. J. 1:569-571

130 DOGS: LABORATORY ANIMAL MANAGEMENT Whitney, J. C. 1967. The pathology of the canine genital tract in false pregnancy. J. Small Anim. Pract. 8:247-263. Whittick, W. G., ed. 1990. Canine Orthopedics, 2d ed. Philadelphia: Lea & Febiger. 936 pp. WHO (World Health Organization) Scientific Group. 1986. Primary immunodeficiency dis- eases. Clin. Immunol. Immunopathol. 40:166-196. Willis, M. B. 1989. Genetics of the Dog. London: H. F. & G. Witherby. 417 pp. Winkelstein, J. A., L. C. Cork, D. E. Griffin, J. W. Griffin, R. J. Adams, and D. L. Price. 1981. Genetically determined deficiency of the third component of complement in the dog. Science 212: 1169- 1170. Winkelstein, J. A., J. P. Johnson, A. J. Swift, F. Ferry, R. Yolken, and L. C. Cork. 1982. Genetically determined deficiency of the third component of complement in the dog: In vitro studies on the complement system and complement-mediated serum activities. J. Immunol. 129:2598-2602. Winkelstein, J. A., J. P. Johnson, K. M. O'Neil, and L. C. Cork. 1986. Dogs deficient in C3. Progr. Allergy 39:159-168. Wi'sniewski, H., A. B. Johnson, C. S. Raine, W. J. Kay, and R. D. Terry. 1970. Senile plaques and cerebral amyloidosis in aged dogs: A histochemical and ultrastructural study. Lab. Invest. 23:287-296. Young, D. M., ed. 1979. Part XV: Skeletal system. Pp. 197-264 in Spontaneous Animal Models of Human Disease, vol. II, E. J. Andrews, B. C. Ward, and N. H. Altman, eds. New York: Academic Press.

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This newly revised edition incorporates the regulatory requirements and improved practices for laboratory animal care that have developed over the past two decades.

The volume covers:

  • Selection of dogs as research models.
  • Design, construction, and maintenance of facilities.
  • Temperature, humidity, food, water, bedding, sanitation, animal identification, record keeping, and transportation.
  • General veterinary care, as well as special care of breeding animals and random-source animals.

Laboratory Animal Management: Dogs examines controversies over proper cage sizes and interpretation of federal requirements for exercise and offers recommendations for researchers. Guidelines are provided on how to recognize and alleviate pain and distress in research dogs and on the sensitive topic of euthanasia.

Laboratory Animal Management: Dogs discusses how to assemble a proper research protocol and how to handle conflicts. Outlined are procedures for institutional animal care and use and committee review. The volume also presents guidelines for handling aging dogs, use of radiation in experiments, and a wide range of other special circumstances.

Thoroughly referenced, this guide will be indispensable to researchers, research administrators, review committees, and others concerned about laboratory dogs.

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