Reptiles used for research and instruction may require surgical procedures, including biopsy, coelomic device implantation, ovariectomy, orchidectomy, and esophogostomy tube placement, to accomplish research goals. Providing veterinary care for unanticipated clinical problems may require
surgical techniques such as amputation, bone or shell fracture repair, and coeliotomy. Although many principles of surgery are common between mammals and reptiles, important differences in anatomy and physiology exist. Veterinarians who provide care for these species should be aware of these differences. Most reptiles undergoing surgery are small and require specific instrumentation and positioning. In addition, because of the wide variety of unique physiologic and anatomic characteristics among snakes, chelonians, and lizards, different techniques may be necessary for different reptiles. This overview describes many common reptile surgery techniques and their application for research purposes or to provide medical care to research subjects.
Little information is available on normal ruminal pH values for domestic sheep (Ovis aries) housed in a research setting and fed a complete pelleted ration. Sheep maintained on pelleted diets undergoing surgical procedures often present with postoperative anorexia and rumen atony. To determine the relationship between diet and postoperative rumen acidosis and associated atony, we studied dietary effects on ruminal pH in an ovine surgical model. Sheep undergoing orthopedic surgical procedures were randomized into 2 diet groups. Group 1 (n = 6) was fed complete pelleted diet during the pre- and postoperative period, and group 2 (n = 6) was fed timothy grass hay exclusively throughout the study. Measures included ruminal pH, ruminal motility, and rate of feed refusal, which was monitored throughout the pre- and postoperative periods. The 2 groups did not differ significantly before surgery, and the ruminal parameters remained largely within normal limits. However, a downward trend in the strength and frequency of rumen contractions was observed in pellet-fed sheep. After surgery, the pellet-fed group showed clinical signs consistent with ruminal acidosis, supported by decreased ruminal motility, anorexia, putrid-smelling ruminal material, and death of ruminal protozoa. Intervention by transfaunation in clinically affected sheep resulted in resolution of signs. Our findings suggest that sheep fed grass hay appear to have a more stable ruminal pH, are less likely to experience anorexia and rumen atony, and thereby exhibit fewer postoperative gastrointestinal complications than do sheep on a pellet diet.
The C57BL/6 mouse strain is used widely for producing transgenic and knockout strains. Sperm motility is extremely low after a freeze–thaw process. Although intracytoplasmic sperm injection (ICSI) can be used to produce embryos from sperm with low or even no motility, its success rate is poor in the C57BL/6 strain. In particular, the survival of C57BL/6 oocytes after ICSI is extremely low compared with that of hybrid strains. We found that the survival percentages of C57BL/6J oocytes (63% and 64%) were lower than those of B6D2F1 oocytes (80% and 80%) when B6D2F1 and C57BL/6J sperm were injected, respectively. For C57BL/6J mice, 87%, 72%, 64%, 56%, and 59% of oocytes survived after ICSI in media containing 61.62, 71.62, 81.62, 91.62, and 101.62 mM NaCl, respectively. In addition, 64%, 81%, and 79% of oocytes survived after ICSI in media with 4.83, 14.83, and 24.83 mM KCl, respectively. Our results suggest that the survival of C57BL/6J oocytes after ICSI is improved by using Na+-deficient and K+-rich media.
Facility planners, IACUCs, veterinary staff, and researchers make choices on water delivery systems for rodents on the basis of cost effectiveness, water quality, risk of malfunction, and potential effect on animal health and welfare. Here we compare biometrics, including weight trends, of newly arrived mice unfamiliar with automated watering; weight trends of weanlings; fecundity of mice; and risk of malfunction among 3 water delivery techniques: water bottle only, combination of automated delivery and water bottle, and automated system only. There was no statistically significant difference among the 3 experimental groups with respect to fecundity, mortality, and delivery malfunction. On the basis of body weight trends, the health and wellbeing of the mice used in these studies were not affected by the water delivery system or housing density after the first week; however, there was a significant difference in the growth rate at 21 to 28 d of age among the 3 groups of pups. The mice receiving both automated delivery and water bottles experienced higher growth rates from 21 to 28 d of age than did the other experimental groups. However, after 35 d of age, weight trends did not differ among the groups. Our results suggest that mice weaned into the same method of water delivery as their respective dams thrive equally well among the 3 tested water delivery systems.
Routine laboratory procedures can be stressful for laboratory animals. We wanted to determine whether human handling of adult rabbits could induce a degree of habituation, reducing stress and facilitating research-related manipulation. To this end, adult New Zealand white rabbits were handled either frequently or minimally. After being handled over 3 wk, these rabbits were evaluated by novel personnel and compared with minimally handled controls. Evaluators subjectively scored the rabbits for their relative compliance or resistance to being scruffed and removed from their cages, being transported to a treatment room, and their behavior at all stages of the exercise. Upon evaluation, handled rabbits scored significantly more compliant than nontreated controls. During evaluation, behaviors that the rabbits displayed when they were approached in their cages and while being handled outside their cages were recorded and compared between study groups. Handled rabbits displayed behavior consistent with a reduction in human-directed fear. This study illustrates the potential for handling to improve compliance in laboratory procedures and reduce fear-related behavior in laboratory rabbits. Such handling could be used to improve rabbit welfare through the reduction of stress and exposure to novel stimuli.
To assess the effect of Xenopus husbandry on oocyte quality for membrane transport physiology experiments, we compared a recirculating-water housing system with a static-water system in a 23-mo study. Two groups of frogs (n = 8) were maintained separately for the entire study: one group was housed in a multiinvestigator centrally managed Xenopus facility, which consists of 33 tanks placed on a shared and recirculating water system; the other group was housed in a satellite facility used by a single investigator and consisting of static tanks placed in a dedicated cold-room. The activity of a heterologously expressed membrane transporter was assessed every 4 to 5 wk for a total of 23 mo. Activity of the mouse cotransporter NKCC1 was assessed through isotopic 86 Rb influx measurements under 2 experimental conditions: stimulation of cotransporter by coinjection of regulatory kinases and by exposure to a hypertonic solution. The results showed a significant difference in the level of ion fluxes under these 2 experimental conditions between the 2 groups of oocytes. During the entire period, oocytes isolated from frogs maintained in the static facility demonstrated consistently robust NKCC1 function, whereas oocytes isolated from frogs maintained in the recirculating facility showed inconsistent and weaker cotransporter function. Furthermore, the oocytes isolated from frogs maintained in the recirculating facility showed significant deterioration during the summer months (April to August), a seasonal variation that was muted in frog oocytes maintained in the static facility.
Identification and eradication of murine fur mite infestations are ongoing challenges faced by many research institutions. Infestations with Myobia musculi and Myocoptes musculinus can lead to animal health problems and may impose unwanted research variables by affecting the immune and physiologic functions of mice. The purpose of this study was to evaluate the utility and efficacy of soiled bedding sentinels in the detection of fur mite infestations in colony mice. Female young-adult CRL:CD1(ICR) mice (n = 140) were exposed over a 12-wk period to various volume percentages of soiled bedding (11%, 20%, 50%, and 100%) from fur-mite–infested animals. Mice were tested every 2 wk with the cellophane tape test to identify the presence of fur mite adults and eggs. At the end of 12 wk, all mice exposed to 11%, 20%, and 50% soiled bedding tested negative for fur mites. One of the 35 mice (3%) receiving 100% soiled bedding tested positive for fur mites at the end of the 12-wk follow-up period. These findings suggest that the use of soiled bedding sentinels for the detection of fur mite infestations in colony mice is unreliable.
This report describes a simple design of noninvasive carbon tube electrodes that facilitates electrocardiography (ECG) in mice during cardiac multimodality preclinical imaging. Both forepaws and the left hindpaw, covered by conductive gel, of mice were placed into the openings of small carbon tubes. Cardiac ECG-gated single-photon emission CT, X-ray CT, and MRI were tested (n = 60) in 20 mice. For all applications, electrodes were used in a warmed multimodality imaging cell. A heart rate of 563 ± 48 bpm was recorded from anesthetized mice regardless of the imaging technique used, with acquisition times ranging from 1 to 2 h.
Threats of nuclear and other radiologic exposures have been increasing, but no countermeasure for acute radiation syndrome has been approved by regulatory authorities. Because of their similarity to humans in regard to physiology and anatomy, we are characterizing Gottingen minipigs as a model to aid the development of radiation countermeasures. Irradiated minipigs exhibit immunosuppression, severe thrombocytopenia, vascular leakage, and acute inflammation. These complications render serial acquisition of blood samples problematic. Vascular access ports (VAP) facilitate serial sampling, but their use often is complicated by infections and fibrin deposition. We demonstrate here the successful use of VAP for multiple blood samplings in irradiated minipigs. Device design and limited postoperative prophylactic antimicrobial therapy before irradiation were key to obtaining serial sampling, reducing swelling, and eliminating infection and skin necrosis at the implantation site. Modifications of previous protocols included the use of polydioxanone sutures instead of silk; eliminating chronic port access; single-use, sterile, antireflux prefilled syringes for flushing; strict aseptic weekly maintenance of the device, and acclimating animals to reduce stress. VAP remained functional in 19 of 20 irradiated animals for as long as 3 mo. The remaining VAP failed due to a small leak in the catheter, leading to clot formation. VAP-related sepsis occurred in 2 minipigs. Blood sampling did not cause detectable stress in nonanesthetized sham-irradiated animals, according to leukograms and clinical signs.
Ischemia of the myocardium can lead to reversible or irreversible injury depending on the severity and duration of the preceding ischemia. Here we compared sevoflurane and isoflurane with particular reference to their hemodynamic effects and ability to modify the effects of acute severe myocardial ischemia and reperfusion on ventricular arrhythmias and mortality in a porcine model of myocardial infarction. Female Large White pigs were premedicated with ketamine, midazolam, and atropine. Propofol was given intravenously for the anesthetic induction, and anesthesia was maintained with isoflurane or sevoflurane. Endovascular, fluoroscopy-guided, coronary procedures were performed to occlude the midleft anterior descending artery by using a coronary angioplasty balloon. After 75 min, the balloon catheter system was withdrawn and the presence of adequate reperfusion flow was verified. The pigs were followed for 2 mo, and overall mortality rate was calculated. The isoflurane group showed lower arterial pressure throughout the procedure, with the difference reaching statistical significance after induction of myocardial ischemia. The ventricular fibrillation rate was higher in isoflurane group (81.3%) than the sevoflurane group (51.7%; relative risk, 1.57 [1.03 to 2.4]). Overall survival was lower in the isoflurane group (75%) than the sevoflurane group (96.4%). In conclusion, in this porcine model of myocardial ischemia and reperfusion, sevoflurane was associated with higher hemodynamic stability and fewer ventricular arrhythmias and mortality than was isoflurane.
To characterize primate arterial waveforms, we prospectively studied 38 bonnet macaques (Macaca radiata; 25 female, 13 ± 4 y). Brachial artery waveforms were recorded from these animals by applanation tonometry and were decomposed into harmonics by using Fourier analysis. The ratio of individual to total harmonic amplitude (H:T) was derived from frequency spectra. Left ventricular (LV) mass, ejection fraction, fractional shortening, septal wall thickness, posterior wall thickness, LV end-diastolic diameter, and LV end-systolic diameter were obtained by echocardiography in all 38 monkeys. Blood pressure was obtained by sphygmomanometry. The fundamental frequency was 2.76 cycles/s. Harmonics ranged from 5 to 14. Indexed LV mass was inversely correlated with third H:T and second H:T but not with systolic or diastolic blood pressure. In addition, the third H:T was inversely correlated with septal wall thickness, posterior wall thickness, and LV end-diastolic diameter, whereas second H:T was inversely correlated with LV end-diastolic diameter. Heart rate was inversely correlated with eighth H:T. On multivariate analysis adjusting for age, gender, weight, and length, only third H:T was an independent predictor of LV mass. Harmonic analysis of arterial waveforms may provide information pertaining to LV mass. Lower H:T ratios (second and third) are related to LV mass, whereas higher H:T (eighth) is related to heart rate.
Our animal care facility has always relied on an animal health team consisting of veterinarians, veterinary care technicians, and husbandry staff to provide a high level of animal care. As our rodent population increased, it became necessary to modify the roles and responsibilities of these staff members to accommodate the program's expansion. To accomplish that modification, we developed a training program that focused primarily on technicians by using a case-management algorithm. To support our technicians, we provided additional training to animal husbandry staff as they assumed the primary role in the initial assessment of the animals' health. After completing the training, technicians made the transition from simply identifying health issues to actually making decisions for treating and euthanizing rodents. This training program empowered all team members and resulted in a staff that could provide consistent, high-quality veterinary care more efficiently.
A 26.6-kg, intact, 9-mo-old female Göttingen minipig was presented for a coronary stent study. Angiography revealed a sinus of Valsalva aneurysm (SVA) in the aortic root that involved both the left and noncoronary sinuses of the heart. Gross histologic examination of the heart revealed 2 regions of aneurysmal formation: one at the ostium to the left main coronary artery, with aortic sinus involvement, and the other at the dorsal aspect of the aortic root involving the noncoronary aortic sinus. With no history of any infectious diseases, and the microscopic findings showing no evidence of necrosis, degeneration, or infection, confirmed that the aneurysmal-like dilation of the sinuses was most likely a congenital anomaly. This case illustrates the diagnosis and comparative findings of a rare cardiac anomaly found in only a few species to date. To our knowledge, antemortem diagnosis of unruptured SVA involving both the left and noncoronary aortic sinuses of the minipig heart has not been reported previously.
Literature on spontaneous primary bone tumors in nonhuman primates is sparse. This case report describes 2 different neoplastic bone lesions in 2 adult cynomolgus monkeys (Macaca fascicularis), including macroscopic, radiographic, histologic, and immunohistochemical findings. In one monkey, a firm mass located at the palatogingival junction of the left rostral maxilla was confirmed to be a peripheral ossifying fibroma in light of its histologic and immunohistochemical characteristics. In another monkey, a lobulated tumor at the right distal femur that radiographically showed moderate radiopacity with splotchy areas of mineralization was confirmed to be a juxtacortical chondrosarcoma on histologic examination. The 2 neoplastic bone lesions revealed rare histologic and immunohistochemical characteristics and contribute to the known tumor spectrum of cynomolgus monkeys.
Mycobacterium infections in nonhuman primates can devastate the colonies and place human handlers at risk. Despite conservative measures to prevent exposure, infections occur even in closed colonies. Here we describe a recent case of M. tuberculosis within a closed colony of rhesus macaques at our Thailand facility and the procedures instituted to prevent subsequent infections. Investigation of the outbreak did not confirm the source of the infection, but even with intensive occupational safety measures in place, human contact remains the most likely possibility.
Simian varicella virus was diagnosed in 2 geriatric rhesus macaques (Macaca mulatta). The macaques presented with typical skin lesions as well as severe thrombocytopenia as a result of infection. Idiopathic thrombocytopenic purpura is a known complication of varicella zoster virus infection in humans; however, this condition has not been reported previously as a complication of SVV infection. This case report discusses the clinical presentation, pathology, and thrombocytopenia of the affected macaques.