The Comparative Medicine (CM) area of the National Institutes of Health (NIH) is a major source of support for research on laboratory animals, training of laboratory animal specialists, and support of shared, regional animal resources. We present a brief history of CM at NIH and the major mechanisms by which it accomplishes its goals in programs located across the United States.

Background and Purpose: Although valuable information has been gained using a rodent partial hepatectomy model to assess liver regeneration, the ability to apply this research to humans remains uncertain. Thus, liver regeneration was assessed in a non-human primate, the rhesus macaque (Macaca mulatta). Methods: One animal underwent 60% hepatectomy, a second animal underwent 30% hepatectomy, and control surgery (cholecystectomy) was performed on two separate animals. Laparoscopic-guided liver biopsy was performed on days 1, 2, 7, 14, and 30 after surgery. Changes in hemoglobin concentration and alanine transaminase activity were assessed, and liver regeneration was evaluated by measuring the expression of Ki-67. Results: All animals survived surgery and laparoscopy. Substantial liver regeneration was induced in the animal that underwent 60% hepatectomy. Excellent tissue specimens were obtained via laparoscopic-assisted liver biopsy. Conclusions: Sixty percent partial hepatectomy in rhesus macaques appears to be an excellent model for the study of hepatocellular regeneration. The procedure was safe, and effectively induced liver regeneration. In addition, laparoscopic-guided liver biopsy allows observation of changes in the liver remnant as regeneration develops, and provides excellent tissue specimens for analysis. Thus, this rhesus macaque partial hepatectomy model will allow further characterization of liver regeneration in a species closer to humans.

Background and Purpose: Rat chromosome 20 is one of special interest because it contains some diabetogenic genes, such as a major histocompatibilitiy complex (MHC)-linked genetic components and quantitative trait loci. We studied rat chromosome 20, using the backcross progeny between BB/Wor and PVG. R23 rats, and confirmed the genetic linkage map by use of another backcross panel. Methods: Backcross panels were done between BB/Wor and PVG. R23 rats, and BN and KZC rats. Length variations of simple sequence length polymorphism markers were analyzed by use of polymerase chain reaction (PCR) analysis. Alleles of RT1-Bb and RT1- Db were analyzed by use of the PCR-restriction fragment length polymorphism method. Genetic maps of rat chromosome 20 were constructed, using the Map Manager computer program. Results: Fifty-two loci were mapped on rat chromosome 20. Genetic length was 57.9 cM, with average spanning of 1.11 cM between markers. The positions of RT1-N1, Tnf, and RT1-Bb into the MHC region were separated and confirmed by results of two backcross panels in our linkage studies. Conclusions: The genetic linkage map of rat chromosome 20 was improved, and was a useful tool for genetic analysis of a diabetogenic gene(s) and for producing MHC congenic strains.

Background and Purpose: The Coxiella burnetii phase-I cellular vaccine is efficacious in humans, imparting nearly complete protection against Q fever. However, this vaccine can also induce sterile abscesses and granulomas at the inoculation site in humans previously sensitized by natural infection or vaccination. To decrease the possibility of vaccinating immune persons, vaccinees are currently screened by skin testing to detect pre-existing Q fever immunity. We developed a model of abscess hypersensitivity in Hartley guinea pigs to assess the likelihood that Q fever vaccines would induce adverse vaccination reactions in previously sensitized individuals. Methods: Guinea pigs (4 to 6/group) were sensitized to C. burnetii by immunization and aerosol challenge, or by intraperitoneal inoculation. Eight weeks later, animals were then vaccinated SC with a Q fever cellular (WCI) or chloroform:methanol residue (CMR) vaccine. Development of adverse reactions at the vaccination site was assessed histologically and by observation of increases in erythema and/or induration. Results: The WCI vaccine caused greater magnitude and duration of erythema and induration at the vaccination sites than did the CMR vaccine. In addition, non-immune guinea pigs developed induration when given WCI, but not CMR vaccine. Conclusions: The CMR vaccine may prove a safe alternative to WCI vaccines for use in individuals unscreened for prior immunity to C. burnetii.

Background and Purpose: Measurement of the biparietal diameter (BPD) by use of B-mode ultrasound provides a useful means for assessment of gestational age and brain growth in humans during pregnancy. Recording of flow velocity waveforms from the umbilical artery, using Doppler ultrasound, is used to assess development of the fetal placental circulation. We sought to measure these ultrasound parameters during normal pregnancy in the guinea pig and develop normative data. Methods: Measurements of BPD were made on 205 fetuses of various gestational ages; 114 fetuses had 2 or more serial studies performed (total n = 474). Results: BPD increased from 0.806cm at 22 to 26 days, to 1.922cm at term (69 days), (y = –0.00043x² + 0.06881x – 0.75941, with an r value of 0.995, where x = days' gestation, y = biparietal diameter [cm]). Umbilical artery flow velocity waveform resistance index (RI) decreased as gestation advanced (y = –0.012x + 1.294 with an r value of 0.887, where x = days gestation, y = RI) reflecting expansion of the placental vascular bed. Conclusions: It is possible to use ultrasound to study pregnancy in the guinea pig. The BPD may be used to estimate gestational age. Resistance to blood flow in the placenta may be assessed using the RI derived from the umbilical artery flow velocity waveform.

Background and Purpose: Left ventricular dysfunction following myocardial infarction is the most important predictor of adverse prognosis. Novel treatment options in infarction require an appropriate experimental model with a standardized, hemodynamically relevant myocardial injury. We evaluated a cryoinjury model in rodents that allows quantitative analysis of systolic and diastolic dysfunction. Methods: Anesthetized, orally intubated, and ventilated Lewis rats (n = 12) underwent sternotomy. Myocardial necrosis was induced by use of a standardized cryolesion to the obtuse margin of the left ventricle, freezing for 3 minutes to –160°C. Left ventricular performance was analyzed at day 120 after cryoinjury. Sham-operated animals (n = 10) served as controls. Results: Cryoinjured animals behaved normally and gained weight up to day 120. Average heart weight of cryoinjured animals significantly exceeded that of controls. Left ventricular systolic pressure and systolic, diastolic, and mean aortic pressures were lower 4 months after cryoinjury, whereas left ventricular end-diastolic pressure was significantly increased. Cryoinjured animals had reduced aortic blood flow, as well as impaired maximal left ventricular dP/dt during aortic occlusion and aortic occlusion-provoked peak systolic pressure. Analysis of maximal rates of isovolumic pressure decrease revealed significant reduction in peak negative dP/dt in cryoinjured animals. Finally, time constants of isovolumic pressure decline were significantly prolonged in cryoinjured animals. Conclusion: Standardized cryothermia induces a myocardial lesion that results in highly reproducible impairment of left ventricular performance 120 days after cryothermia. The model is ideally suited to test novel therapeutic strategies for myocardial dysfunction.

Background and Purpose: The current safety standards for lasers operating in the 1,400- to 2,000-nanometer (nm) wavelength region are based on only a few observations at specific wavelengths. On the basis of experimental results conducted with Yorkshire pigs (Sus scrofa domestica), these standards may not accurately reflect the potential for laser injury when humans are exposed to these wavelengths. It is our belief that one of the damage mechanisms involved in these laser injuries results from energy absorption by skin pigmentation (melanin), and a more highly pigmented animal model, the Yucatan hairless minipig, may be a more suitable subject for laser exposure studies. Methods: Skin specimens were collected from Yorkshire pigs and Yucatan minipigs for histologic examination, and the thickness of the epidermis was measured. Epidermal thickness of human skin also was determined, and a qualitative assessment of the melanin content in the epidermal layers was conducted. Results: Mean ± SD thicknesses of the Yucatan minipig flank and dorsal neck epidermis were 68 ± μ34 and 68 ± μ25 m, respectively. Thicknesses of the Yucatan minipig skin were closely comparable to the thicknesses of human epidermis from the face (68 ± 26 μm), neck (65 ± 24 μm) and arms (68 ± 21 μm). The Yorkshire pig lacks substantial melanin in the epidermis, whereas the skin of the Yucatan minipig is more similar to that of humans. Conclusion: On the basis of epidermal skin thickness measurements and melanin assessment, the flank and dorsal neck of the Yucatan minipig are better suited to laser injury studies than are the Yorkshire pig models of human skin.

The effects of long-term (3-day) infusion of nonphysiologic solutions into brain parenchyma were inves- tigated in male Fischer (F344) 344 rats. Two weeks prior to infusion, a guide cannula was placed into the striatum, substantia nigra, or hippocampus. Solutions were infused continually for 3 days at flow rates of 0.03 (129.6 μl total) or 0.10 (432 μl total) l/min. Four days after infusion, rats were euthanized and the brain was removed and processed for histologic evaluation. Rats that received cannula implants alone had the usual mechanical damage induced by implantation of the cannula. The brain regions that received 0.9% saline, pH 5.0 or pH 9.0 buffer at the two aforementioned flow rates had only minor evidence of tissue damage adjacent to the infusion site that was similar to that attributable to mechanical damage from the cannula implants. Brain tissue infused with distilled water or 1.8% saline also had modest effects of the solutions similar to the usual mechanical damage induced by the infusion cannulae. In contrast, contamination of the infusion sites was seen to induce inflammation. Data from these studies support the hypothesis that nonphysiologic solutions can be used to deliver compounds into brain parenchyma, without the infusion solutions themselves causing excess damage to brain tissue.

Objective: The aim of the study reported here was to set up a method for echocardiography (EC) and abdominal sonography and to obtain EC reference values for left ventricular (LV) morphology and function and sonographic abdominal aortic morphology, function, and flow values in conscious, unsedated Gottingen minipigs. Methods: Applying a standardized investigation procedure, the following parameters were measured by use of Mmode EC, color-coded Doppler imaging, and B-mode sonography, or were calculated, in 58 female minipigs: LV enddiastolic and end-systolic diameter, interventricular septum thickness, LV caudal wall thickness, LV end-diastolic volume and end-systolic volume, fractional shortening, ejection fraction, and percentage of thickening of interventricular septum and LV caudal wall. In addition, morphology, pulsatility, flow values, and flow patterns in the abdominal aorta were recorded or calculated during abdominal sonography and color-coded Doppler imaging. Results: Variable EC values were obtained due to individual variations of motor activity. Variation could be reduced by accustoming the animals to a standardized investigation procedure. Reference values could be obtained for EC, partially indicating clear correlation with body weight. Color-coded Doppler and Doppler spectra did not indicate flow disturbances in large arterial abdominal vessels. Conclusions: Results indicate that handling during EC and sonography can cause discomfort in unsedated minipigs that may interfere with recording of valid reference values for functional cardiac parameters in young animals. Accustoming the animals to a standardized investigation procedure reduces stress to a satisfactory level and enables data recording. Thus the minipig is considered suitable for assessment of cardiovascular parameters in experimental or toxicologic studies.

Background and Purpose: We characterized abnormalities of carbohydrate and lipid metabolism and determined whether those metabolic abnormalities are associated with extremity lesions in California mice (Peromyscus californicus). Methods: Blood samples were evaluated for glucose, cholesterol, triglyceride, and insulin concentrations. Necropsy and histologic evaluation were done on selected mice, including staining pancreatic sections for insulin. Physical examinations also were performed. Results: California mice were found to have Type 2 diabetes mellitus (T2DM). Sections of pancreas from diabetic and prediabetic mice had pathologic changes consistent with T2DM. After six months of feeding a low-fat diet, mice were normoglycemic, normotriglyceridemic, and normocholesterolemic. Some mice remained hyperinsulinemic. Traumatic lesions were not associated with T2DM. Conclusions: California mice develop diet-related T2DM when fed a diet containing 25.8% kcal from fat. California mice may be a useful animal model of human T2DM, and traumatic lesions result from housing California mice in multiple male groups.

Background and purpose: Self-injurious behavior (SIB) affects 0.8 to 10% of individually housed non-human primates, and is a substantial threat to their health and well being. The potential for SIB to involve multiple neurotransmitters and the complex variations in response to external stressors complicate case management. Modulation of the adrenergic system by use of guanfacine, an α_2A -adrenergic receptor agonist, was assessed as a novel therapeutic strategy for SIB. Methods: The efficacy of guanfacine against SIB was evaluated in 11 self-biting episodes among two rhesus macaques (Macaca mulatta) and one baboon (Papio cynocephalus anubis). Affected animals were given guanfacine IM or PO at 0.5 mg/kg of body weight twice daily (rhesus) or 0.3 mg/kg (baboon) for 5 to 10 days, followed by gradual reduction of the dose to 0.25 mg/kg (rhesus) or 0.15 mg/kg (baboon) once daily over an average of 33 days. Results: The 0.5 mg/kg twice daily regimen of guanfacine halted all self-biting, whereas reducing the dose to 0.25 mg/kg given twice daily or 0.5 mg/kg given once daily resulted in reversion to self-biting in four of the 11 episodes. Recurrence was controlled by returning to twice daily 0.5 mg/kg dosing for one aggressive episode, and resolved in the three milder episodes without dose or frequency being increased. Self-biting after discontinuation of therapy recurred six times over five years in case 1, three times over 1.5 years in case 2, and three times over one year in case 3. Clinical assessment suggested that guanfacine therapy decreased agitation without overt side effects associated with α₂ -agonists, such as profound sedation. Conclusion: The mechanism for guanfacine inhibition of self-biting is unclear, but could result from strengthening of prefrontal cortex inhibitory functions. Guanfacine therapy provides an effective psychological stabilizing tool that alleviates self-biting, and provides time to assess and address external stressors and triggers.

Mousepox was identified in a single mouse-holding room in early 1999 after a group of 20 CAF1/Hsd mice were inoculated SC with a killed murine spindle cell tumor line, S1509A. The cell line had been used without complications multiple times and was determined to be free of viral contamination on the basis of results of mouse antibody production testing. Of the 20 mice inoculated, 12 mice died by postinoculation day 8. Severe lymphoid and hepatic necrosis was observed in select mice subjected to histologic examination. Ballooning degeneration of epithelial cells with intracytoplasmic eosinophilic inclusion bodies was observed in the skin overlying the inoculation site of the single mouse from which this tissue site was evaluated. Presence of ectromelia virus was confirmed by use of immunohistochemical and polymerase chain reaction analyses, and the virus was isolated after serum, pooled from 5 of the index cases, was inoculated into an immune-naive mouse. Investigation into the source of virus contamination included inoculating mice with aliquots of various S1509A freeze dates; chemically defined media and supplements, including fetal bovine serum; and two lots of pooled commercial mouse sera, after heat inactivation at 56°C for 30 minutes used as a medium supplement. One lot of pooled commercial mouse serum was identified as the source of ectromelia virus. This lot of serum was inadvertently used to feed S1509A cells that were subsequently inoculated into mice. We determined that the contaminated serum, which was purchased in late 1998, originated from China. The serum was imported into the United States as a batch of 43 L in early 1995. The serum was blended into a single lot and filtered (0.μ2 m) before distribution to major suppliers throughout the country. The serum was sold or further processed to obtain a variety of serum-derived products. Because murine serum is generally sold in small aliquots (10 to 50 ml), we speculate that several thousand aliquots may have been derived from this batch of serum and, if inoculated into mice, would likely result in additional mousepox outbreaks.

Background and Purpose: Natural infection of research mice with enterohepatic Helicobacter spp. is common and may confound experimental studies from intercurrent disease. We evaluated a protocol of dirty bedding exposure for transmission of Helicobacter infection from colony mice to female Tac:(SW)fBR sentinel mice over 6 months. Methods: Cecal scrapings from culled colony mice and associated sentinel mice were tested for H. hepaticus, H. rodentium, and H. bilis using polymerase chain reaction analysis (PCR). These results were correlated with the results of sentinel serum IgG responses measured by ELISA. Results: In 9 colony rooms, 43 of 45 mice were infected with H. hepaticus ; in 14 rooms, 58 of 70 mice were infected with H. rodentium ; and in 2 rooms, 2 of 10 mice were infected with H. bilis. Concurrence of Helicobacter infection between colony and sentinel mice was 82% for H. hepaticus, 88% for H. rodentium, and 94% for H. bilis. Concurrence of Helicobacter infection status of sentinel cagemates was 98% for H. hepaticus, 86% for H. rodentium, and 95% for H. bilis. Fecal samples pooled by sentinel cage had positive PCR results for H. hepaticus and H. rodentium at 1 month in 60 and 44%, respectively, of the cages that contained test-positive mice at necropsy (6 months). By 3 months, detection rates were 100 and 81% for H. hepaticus and H. rodentium, respectively, and H. bilis was not detected until 4 months. Newly acquired infections with H. rodentium and H. bilis were evident throughout the 6-month study period. Seroconversion was coincident with positive PCR results in sentinel mice, and serum IgG values continued to increase until necropsy. The serum IgG ELISA was 98 to 100% sensitive, but was low in specificity (34 to 44%), most likely attributable to common coinfection with H. hepaticus and H. rodentium. Conclusion: Sentinel mice acquire infection with Helicobacter spp. through dirty bedding exposure. Combined use of PCR analysis and serologic testing of sentinel mice was predictive of Helicobacter infection status of mouse colonies used for biomedical research.

Background and Purpose: Efficient methods for detection and elimination of Helicobacter spp. infections are needed to facilitate the development of Helicobacter -free mouse colonies. We developed an inexpensive, high-throughput method for preparation of fecal DNA for Helicobacter polymerase chain reaction (PCR) assays. Methods: Fecal DNA was prepared by heating fecal pellets to 95°C for 10 minutes in an alkaline solution, then adjusting the pH by addition of Tris buffer. This solution is used for PCR assays without purification of DNA. We then tested fostering as a method of generating Helicobacter-free mice. Litters born to Helicobacter-positive dams were transferred to Helicobacter -negative foster dams on the first day of life. Results: Fostered pups tested Helicobacter negative up to 89 days of age, whereas pups raised by Helicobacter positive dams were all test positive by 19 days of age. Conclusion: These simple methods provide an efficient system for the development of Helicobacter-free mouse colonies.

A 30-year-old male Sumatran orangutan ( Pongo pygmaeus ) presented with signs of depression, leth- argy, anorexia, and diarrhea that progressed to acute colic. Exploratory laparotomy revealed fibrinopurulent peri- tonitis and 50 cm of devitalized small intestine. The surgically resected small intestine contained several mucosal diverticula along the mesenteric attachment; one had ruptured, resulting in peritonitis. Fifteen days after surgery, the orangutan's abdominal incision dehisced. Repeated laparotomy revealed dehiscence of the distal intestinal anastomosis site, as well as extensive adhesions and purulent exudate. The defect was repaired, and the abdomen was extensively irrigated and closed, but the animal died within 24 hours. To our knowledge, this is the first report of diverticulitis in a great ape. Diverticulosis should be considered in the differential diagnosis for great apes that present with signs of depression, lethargy, anorexia, and/or diarrhea.