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Carbon dioxide euthanasia is an established method for the termination of small laboratory animals. It has also been employed by the authors in neurobiological research on the post-mortem glutamate concentration in the structures of rat brains. The following investigations were aimed at optimizing the termination procedure based on the CO2 saturation rate of the inhaled air. Two rates of CO2 flow were applied, and the higher one significantly augmented the glutamate level in the hippocampus and cerebellum. The relationship between this finding and signs of central fear reaction is discussed. The authors conclude that lower rather than higher CO2 flow in euthanasia procedures is gentler and is therefore preferable for use with laboratory animals.

Objective: To determine the severity and distribution of structural changes in the brains of adult sheep stunned by penetrating captive bolt., Procedure: The unconstrained heads of ten, anaesthetised, unhorned, 2-year-old Merino sheep were impacted at the summit of the head with a penetrating captive bolt pistol. Six sheep were ventilated and four received no respiratory support. Two hours after impact, brains from the six ventilated sheep were perfusion-fixed with 4% paraformaldehyde. Sixteen whole, serial coronal sections from each brain were stained with haematoxylin and eosin and immunohistochemically for amyloid precursor protein, a sensitive marker of axonal and neuronal reaction in the brain after trauma. Pathological changes in these brains were then quantified by morphometric analysis., Results: Structural change in all impacted brains was a mixture of focal injury around the wound track and more widely distributed damage in the cerebral hemispheres, cerebellum and brainstem, but varied considerably in severity between individual sheep. All nonventilated sheep died rapidly following respiratory arrest., Conclusions: After penetrating captive bolt stunning, damage to the central reticular formation, axonal connections, and the cortical mantle is the likely reason for failure of respiratory control and traumatic loss of consciousness.

A procedure has been developed and validated for measuring the concentration of pentobarbital residues in dry, extruded animal feed in the range of 3-200 ng/g (ppb) with an estimated limit of quantitation of 2 ppb. The method was developed for surveillance purposes: to measure the concentration of euthanizing agent which might be present in feeds incorporating rendered products which themselves might include some fraction of euthanized animals. A previously published qualitative procedure was modified by adding isotopically labelled pentobarbital as an internal standard. Dry feed was ground and extracted with methanol. The extract was loaded on a mixed-mode (C-18, anion exchange) solid-phase extraction cartridge designed for barbiturate residues. Pentobarbital was eluted and derivatized for gas chromatography/mass spectrometry in positive ion chemical ionization mode. Quantitation was based on the ratio of dimethyl-pentobarbital MH+ (m/z 255) vs dimethyl-pentobarbital-d(5) (m/z 260) in standards and extracts. Accuracy ranged from 112% at 3 ppb to 96% at 200 ppb, with relative standard deviations ranging from 4% at 3 ppb to 2% at 200 ppb.

Objective: To determine causes of death or reasons for euthanasia in a population of military working dogs., Design: Retrospective study., Animals: 927 military working dogs., Procedure: Records of all military working dogs that died during the period from 1993 to 1996 were evaluated for cause of death or reason for euthanasia by review of necropsy and histopathology reports, death certificates, and daily clinical treatment sheets. A single primary cause of death or euthanasia was determined., Results: Although sexually intact male dogs were more numerous in the study population, castrated male dogs typically lived longer than spayed females or sexually intact males. Leading causes of death or euthanasia (76.3% of all dogs) were appendicular degenerative joint disease, neoplasia, spinal cord disease, nonspecific geriatric decline, and gastric dilatation-volvulus. Compared with German Shepherd Dogs, Belgian Shepherd Dogs were at increased risk for death attributable to neoplasia, behavior, and respiratory tract disease. German Shepherd Dogs had nearly twice the risk for death associated with spinal cord diseases, compared with Belgian Shepherd Dogs., Conclusions and Clinical Relevance: For most military working dogs, death or euthanasia results from a few diseases commonly associated with advanced age. Some breed differences in risk for these diseases may exist, which clinicians should consider in the procurement and long-term management of these dogs.

A retrospective study was carried out of 224 horses operated for strangulating small intestine obstructions. Fifty-four horses were euthanized and 5 horses died during surgery which means that 165 (73%) were allowed to recover. Of these, 53 horses were euthanized or died in the clinic and 112 (50%) were discharged from the hospital. Of 90 horses available for follow-up 1 year postoperatively, 76 (84%) were still alive. The most important causes of death or reasons for euthanasia in the direct post-operative period were post-operative paralytic ileus, (adhesive) peritonitis and intra-abdominal haemorrhage. After discharge from the hospital the reasons were (adhesive) peritonitis and (recurrent) colic. Of the horses which survived for at least 1 year, 16% sometimes suffered from colic, 12% experienced problems with incisional woundhealing and 4% suffered from jugular vein thrombosis. All were in good or reasonable condition and 88% performed at (approximately) the same level as before the operation. The type of surgical intervention (i.e. enterotomy, enterectomy) did not significantly influence the outcome of surgery, whereas the type of anastomosis did. End-to-end jejunojejunostomy had a better prognosis than side-to-side jejunocaecostomy. It was concluded that strangulating obstructions of the small intestine still carry a poor to guarded prognosis. Mortality was highest in the direct peri-operative period. Once discharged from hospital, prognosis can be considered to be fair to good. Attempts to improve outcome should be directed at a better handling of the ileal stump during surgery and at the prevention of post-operative ileus and the formation of adhesions.

A 9-year-old, female guinea pig was diagnosed with a subcutaneous fibrosarcoma overlying the ventral thorax. The mass was invasive within the subcutis without extension into surrounding structures or organ systems. Other findings included cystic ovaries, renal cortical cysts, nodular hyperplasia of the liver, and myocardial fibrosis with pulmonary hypertension.

An 8-year-old, female llama was evaluated for nonhealing, ulcerative, cutaneous lesions, which also involved the mammary gland. Biopsies of the lesions distant from and within the mammary gland area revealed an aggressive carcinoma. The tumor was confirmed at necropsy to be a mammary gland adenocarcinoma with cutaneous metastasis.

Equine protozoal myeloencephalitis is a common neurologic disease of horses in the Americas usually caused by Sarcocystis neurona. To date, the disease has not been induced in horses using characterized sporocysts from Didelphis virginiana, the definitive host. S. neurona sporocysts from 15 naturally infected opossums were fed to horses seronegative for antibodies against S. neurona. Eight horses were given 5x10(5) sporocysts daily for 7 days. Horses were examined for abnormal clinical signs, and blood and cerebrospinal fluid were harvested at intervals for 90 days after the first day of challenge and analyzed both qualitatively (western blot) and quantitatively (anti-17kDa) for anti-S. neurona IgG. Four of the challenged horses were given dexamethasone (0.1mg/kg orally once daily) for the duration of the experiment. All challenged horses immunoconverted against S. neurona in blood within 32 days of challenge and in CSF within 61 days. There was a trend (P = 0.057) for horses given dexamethasone to immunoconvert earlier than horses that were not immunosuppressed. Anti-17kDa was detected in the CSF of all challenged horses by day 61. This response was statistically greater at day 32 in horses given dexamethasone. Control horses remained seronegative throughout the period in which all challenged horses converted. One control horse immunoconverted in blood at day 75 and in CSF at day 89. Signs of neurologic disease were mild to equivocal in challenged horses. Horses given dexamethasone had more severe signs of limb weakness than did horses not given dexamethasone; however, we could not determine whether these signs were due to spinal cord disease or to effects of systemic illness. At necropsy, mild-moderate multifocal gliosis and neurophagia were found histologically in the spinal cords of 7/8 challenged horses. No organisms were seen either in routinely processed sections or by immunohistochemistry. Although neurologic disease comparable to naturally occurring equine protozoal myeloencephalitis (EPM) was not produced, we had clear evidence of an immune response to challenge both systemically and in the CNS. Broad immunosuppression with dexamethasone did not increase the severity of histologic changes in the CNS of challenged horses. Future work must focus on defining the factors that govern progression of inapparent S. neurona infection to EPM.

A 4-year-old castrated male mixed-breed dog with a history of coccidioidomycosis was referred for evaluation of abdominal and pleural effusion. Results of radiography, ultrasonography, cytologic evaluation of thoracic fluid, and serologic testing supported a diagnosis of constrictive pericarditis secondary to infection with Coccidioides immitis. Aggressive treatment for presumptive coccidioidomycosis was begun, but the dog's condition continued to deteriorate, and the dog was euthanatized. At necropsy, the pericardium was thicker than normal and fibrotic and adhered to the epicardium. Microscopically, the pericardium and 1 section of epicardium contained lymphoplasmacytic infiltrates with a few macrophages and neutrophils. Coccidioides immitis was cultured from pericardial fluid. A search of records from the Arizona Veterinary Diagnostic Laboratory for 1988 through 1998 revealed that of 46 dogs in which a diagnosis of coccidioidomycosis was confirmed at necropsy, 13 had involvement of the heart or pericardium.

The issue of euthanasia is unique to veterinary clinical studies evaluating survival time. The decision to euthanize an animal is based on several factors including the health of the animal but also age and cost of treatment. The literature shows inconsistent methods used to account for observations from euthanized animals. Also, over 50% and up to 100% of animals in many studies have been euthanized. Our study illustrates the effects of different methods of accounting for observations from euthanized animals in survival analysis. Three data sets with different proportions of outcomes (alive, lost-to-follow-up, dead due to disease of interest, dead due to other disease, euthanized due to disease of interest, euthanized due to other disease) were used. Each data set was stratified according to treatment or a group characteristic (e.g. tumor type). Our methods for accounting for observations from euthanized animals were established from methods used in the literature and included right-censoring. Kaplan-Meier product-limit survival-function estimation was performed on each data set. Different methods resulted in inconsistent conclusions of significant differences between strata. At times, the ranking of the estimates of median survival time for strata was reversed. Right-censoring and use of Kaplan-Meier methods is inappropriate to evaluate observations from euthanized animals because censoring of such observations is informative. The current methods used by clinical investigators are inadequate to measure survival time reliably.

Transmissible spongiform encephalopathies affect a number of mammalian species. The most common spongiform encephalopathies are scrapie in sheep and Bovine Spongiform Encephalopathy (BSE) in cattle. Feline Spongiform Encephalopathy (FSE) is a related disorder in domestic cats. Because of the link between BSE and FSE, cats are put on a par with cattle, in terms of politics and regulations. In the Netherlands, when a case of BSE is found on a farm, not only the ruminants, but also the cats are taken away for post-mortem examination. So far, the cats examined have always been negative for FSE. There are no scientific reasons for destroying the cats on farms where BSE has been found.

An adult, wild-caught, female prairie falcon (Falco mexicanus) was presented with the chief complaint of anorexia. Radiographic findings included increased densities within the air sacs, and coelomic endoscopy revealed numerous slender worms within the air sacs and on the serosal surfaces of the ovary, oviduct, liver, proventriculus, and ventriculus. The bird seemed to improve for a short period of time with antiparasitic therapy (ivermectin and fenbendazole) and supportive care. Twenty-one days after initial presentation, the bird became recumbent with increasing pelvic limb neurologic deficits and was euthanized. On histopathologic examination, mature nematodes and larvated eggs identified as Serratospiculoides amaculata were found within the subdural space of the distal thoracolumbar and synsacral spinal cord and within the coelomic cavity. This case suggests that S. amaculata can cause clinically significant lesions in its falconiform host with potentially fatal results.