Method for determination of pentobarbital in dry dog food by gas chromatography/mass spectrometry.

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.

Short-and long-term evaluation of surgical treatment of strangulating obstructions of the small intestine in horses: a review of 224 cases.

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.

Causes of death or reasons for euthanasia in military working dogs: 927 cases (1993-1996).

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.

Subcutaneous fibrosarcoma in an aged guinea pig.

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.

Cutaneous metastases of a mammary carcinoma in a llama.

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.

The effects of different methods of accounting for observations from euthanized animals in survival analysis.

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.

Use of sevoflurane for anesthetic management of horses during thoracotomy.

OBJECTIVE: To evaluate sevoflurane as an inhalation anesthetic for thoracotomy in horses. ANIMALS: 18 horses between 2 and 15 years old. PROCEDURE: 4 horses were used to develop surgical techniques and were euthanatized at the end of the procedure. The remaining 14 horses were selected, because they had an episode of bleeding from their lungs during strenuous exercise. General anesthesia was induced with xylazine (1.0 mg/kg of body weight, IV) followed by ketamine (2.0 mg/kg, IV). Anesthesia was maintained with sevoflurane in oxygen delivered via a circle anesthetic breathing circuit. Ventilation was controlled to maintain PaCO2 at approximately 45 mm Hg. Neuromuscular blocking drugs (succinylcholine or atracurium) were administered to eliminate spontaneous breathing efforts and to facilitate surgery. Cardiovascular performance was monitored and supported as indicated. RESULTS: 2 of the 14 horses not euthanatized died as a result of ventricular fibrillation. Mean (+/- SD) duration of anesthesia was 304.9 +/- 64.1 minutes for horses that survived and 216.7 +/- 85.5 minutes for horses that were euthanatized or died. Our subjective opinion was that sevoflurane afforded good control of anesthetic depth during induction, maintenance, and recovery. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of sevoflurane together with neuromuscular blocking drugs provides stable and easily controllable anesthetic management of horses for elective thoracotomy and cardiac manipulation.

Meningeal osteosarcoma in a dog.

A meningeal osteosarcoma was diagnosed in a dog displaying neurologic signs compatible with a space-occupying cerebellar lesion. Gross lesions, restricted to the brain, consisted of a solitary, compressive mass attached to the dura mater overlying the left cerebellum. The mass was composed of single and multinucleated, atypical polygonal cells that lined or rested within lacuna surrounded by eosinophilic, mineralized matrix. The matrical component stained dark green-yellow to blue with Movat's pentachrome stain, deep blue to red with Heidenhain aniline blue stain, and brown-black with Von Kossa stain. Results of these stains were interpreted as tumor osteoid. Foci of dural mineralization and osseous metaplasia were present at the point of tumor attachment. The microscopic observations were interpreted as an osteosarcoma of extraskeletal origin. To our knowledge, these findings represent the first documented case of a meningeal osteosarcoma in a domestic animal species.

Anaemia, diarrhoea and opportunistic infections in Fell ponies.

This report summarises clinical and pathological observations on Fell pony foals with a range of signs that included ill thrift, anaemia, respiratory infection, glossal hyperkeratosis and diarrhoea. Some of the foals had normochromic, normocytic anaemia and some had low levels of plasma proteins, including immunoglobulin G. Antibiotic and supportive treatment was ineffective and all affected foals died or were killed on humane grounds. Postmortem examination of 12 foals and tissues from 2 other foals revealed a range of lesions that included glossal hyperkeratosis, typhlocolitis, intestinal cryptosporidiosis, granulomatous enteritis, proliferative and necrotising bronchiolitis consistent with adenovirus infection; lesions similar to those in the respiratory tract were present in the salivary gland and pancreas of individual foals. Lymphoid tissue was judged to be smaller than expected. These observations suggest the possibility of opportunistic infections secondary to some form of undefined immunocompromised state.

Surgical treatment of 45 horses affected by squamous cell carcinoma of the penis and prepuce.

The medical records of 45 horses treated for suspected squamous cell carcinoma of the penis and/or prepuce were reviewed. The age of 40 horses was known, and these had a mean age of 17.4 years. The duration of neoplasia was known for only 3 of the 45 horses. The results of histological evaluation of lesions, available for 35 horses, confirmed that the diseased tissue was squamous cell carcinoma. The location of gross neoplastic lesions was recorded for 43 horses; the glans penis was involved in 24 horses, the body of the penis or the inner lamina of the preputial fold in 27 horses, and the external fold of the prepuce in 10 horses. Surgical treatments of these horses included phallectomy (penile amputation) in 35 horses, segmental posthectomy in 4 horses, phallectomy plus segmental posthectomy in 2 horses, and en bloc resection of the penis, prepuce and superficial inguinal lymph nodes with penile retroversion in 4 cases. Short-term complications in the immediate postoperative period included preputial oedema and haemorrhage at the end of urination. One horse developed acute urinary retention because of severe urethral oedema. Long-term (>1 year) follow-up information was obtained for 31 horses. Neoplasia of the penis and/or prepuce recurred in 6 of these 31 horses (19%), and in 5 of these the recurrence necessitated euthanasia of the horse.

Isolation and characterization of an antigenically distinct 68-kd protein from nonviral intracytoplasmic inclusions in Boa constrictors chronically infected with the inclusion body disease virus (IBDV: Retroviridae).

The relationship between a retroviral infection and the development of nonviral intracytoplasmic inclusion bodies was studied in a Boa constrictor model. Twelve juvenile age- and size-matched inclusion body disease (IBD)-negative boas were randomly divided into three groups. Each group was inoculated intraperitoneally with 1 ml of an IBD virus (IBDV)-infected liver homogenate or 1 ml of normal boa liver homogenate (sham-inoculated control) or was left untreated. All boas were monitored for development of IBD by daily examination and serial liver biopsy over 1 year. The 4 IBDV-inoculated boas became IBDV and inclusion positive by 10 weeks postinoculation. The average size and density of inclusion bodies increased with the duration of infection. Ultrastructurally, inclusion bodies <2 microm in diameter consisted of intracytoplasmic aggregates of granular electron-dense material that were not membrane limited. Larger inclusions (3-6 microm in diameter) were characterized as membrane-bound aggregates of amorphous to granular electron-dense material admixed with membranelike fragments. The sham-inoculated and untreated control snakes did not become inclusion or IBDV positive. Direct comparison of the protein electrophoretograms of IBDV-infected and normal boa tissues demonstrated a prominent 68-kd protein band unique to infected inclusion-positive tissues. Monoclonal antibodies directed against the 68-kd protein band specifically labeled inclusion bodies. The results of this study demonstrate that IBD inclusions represent an intracytoplasmic accumulation of an antigenically distinct IBDV-associated protein.

Progression of an orbital T-cell rich B-cell lymphoma to a B-cell lymphoma in a dog.

An 11-year-old Shetland Sheepdog was presented for exophthalmos caused by a locally extensive, poorly defined mass located behind the right eye. The primary orbital mass was identified by light microscopy and immunohistochemistry as a T-cell rich B-cell lymphoma (TCRBCL) composed predominantly of BLA.36-positive large neoplastic lymphoid cells admixed with fewer CD3- and CD79a-positive small lymphocytes. The dog was treated for lymphoma, but 6 months after presentation it was euthanatized for suspected hepatic and gastrointestinal metastasis. Gross findings revealed an enlarged liver with multiple well-demarcated, randomly distributed 0.1-1.5-cm white nodules, five firm white submucosal jejunal nodules, and ileocecal, mediastinal, and hilar lymphadenopathy. Metastatic liver lesions consisted of sheets of monomorphic large neoplastic lymphoid cells that effaced and expanded portal and centrilobular zones. These cells were morphologically similar to the large neoplastic cells of the original orbital tumor and were CD3-negative and variably BLA.36-positive, consistent with B-cell lineage. Similar cells comprised the jejunal nodules and effaced the lymph nodes. The progression of TCRBCL to a diffuse B-cell lymphoma in this case is consistent with reported human cases and has not been previously reported in the dog.

Leukoencephalitis and vasculitis with perivascular demyelination in a Weimaraner dog.

Aseptic and noninfectious diseases of the central nervous system are being recognized with increasing frequency. After multiple episodes of neurologic illness, this 7-year-old Weimaraner dog was euthanatized and submitted for postmortem examination. Lesions in the central nervous system were found mainly in the white matter of the cerebral cortex and cervical spinal cord and represented acute and more chronic injury. Necrotizing vasculitis with fibrinoid change and a marked neutrophilic infiltrate dominated the acute lesions. More chronic changes consisted of perivascular demyelination and accumulation of foamy macrophages with positive staining for myelin. An immune-complex (Arthus-type) vasculitis is suspected.

Mediastinal plasma cell tumor in a sheep.

An extramedullary plasmacytoma was found in a 10-year-old sheep. The tumor involved the mediastinum, where a 25 x 15 x 10-cm encapsulated mass was found. The lungs had multiple metastases ranging from 0.5 to 2 cm in diameter, and the portal vein contained a 10-cm-long mass. The cytologic and histopathologic analyses were consistent with a moderately differentiated plasmacytoma. The immunophenotype of the tumor cells was lambda light chain IgG+, CD79a-, and CD3-. Occasional granulomas were observed at the periphery of the mediastinal and pulmonary tumors. Microbiologic culture yielded growth of Corynebacterium from these granulomas. This is the first report of plasmacytoma in sheep. The tumor most likely arose from mediastinal lymph nodes and metastasized to the lungs and portal vein.

Marginal siderosis and degenerative myelopathy: a manifestation of chronic subarachnoid hemorrhage in a horse with a myxopapillary ependymoma.

Marginal siderosis is recognized in humans as an uncommon clinicopathologic entity characterized by degeneration of neural tissue at the surface of the brain and spinal cord, in association with the accumulation of hemosiderin, and resulting from chronic subarachnoid hemorrhage. The sources of hemorrhage are various and include neoplasms, malformations, cysts, and vasculopathy. Marginal siderosis of the spinal cord due to a myxopapillary ependymoma was diagnosed in a 19-year-old Dutch Warm Blood horse with clinical signs of myelopathy. There is only one previous report of marginal siderosis in the veterinary literature, also in a horse with clinical myelopathy.

Anti-plakin and desmoglein autoantibodies in a dog with pemphigus vulgaris.

Paraneoplastic pemphigus was suspected in a 14-year-old Labrador retriever because of mucocutaneous erosions, microscopic suprabasal acantholysis, and keratinocyte apoptosis. In this patient, circulating IgG autoantibodies recognized plakin (envoplakin, periplakin) and desmoglein (desmoglein-1 and -3) antigens. Necropsy, however, failed to confirm the concurrent existence of hematopoietic or solid neoplasia. The diagnosis of pemphigus vulgaris therefore was proposed. This study illustrates that such a combination of clinicopathologic lesions and plakin/desmoglein-specific autoantibodies is not restricted to canine paraneoplastic pemphigus but can also be detected in another form of suprabasal pemphigus.

Mammary adenocarcinoma in a male squirrel monkey (Saimiri sciureus).

A nodule was identified within the right mammary gland of a 16-year-old male squirrel monkey (Saimiri sciureus). The mass was excised and diagnosed as a mammary adenocarcinoma. The monkey developed congestive heart failure 1.5 years later and was euthanatized. At necropsy, a subcutaneous mass was found in the right axillary region. Histologically, the mass was identified as a lymph node whose architecture was effaced by neoplastic cells resembling those of the mammary tumor. Metastasis to internal organs was not observed. This is the first reported case of a mammary tumor in a New World primate and the only known case of mammary cancer in a male nonhuman primate.

Squamous cell carcinoma in an American bison (Bison bison bison).

The following case report is, to the best of our knowledge, the first documentation of a squamous cell carcinoma in a full-blooded American bison (Bison bison bison). This female bison developed an ulcerated lesion on the left flank, at the site of a brand, in the summer of her eighth year. In the winter of her ninth year, as the lesion continued to grow, a biopsy was performed, and a diagnosis of squamous cell carcinoma was made. Ultimately, there was metastasis to the liver, a lung, and a prefemoral lymph node.

Multiple myeloma in cats: variable presentation with different immunoglobulin isotypes in two cats.

Multiple myeloma was diagnosed in two cats with monoclonal hyperglobulinemia, proteinuria, and plasma cell proliferations in bone marrow. An immunoglobulin G-producing myeloma occurred in the vertebral bone marrow of one cat, and twice responded to surgical reduction followed by a combination of local irradiation and chemotherapy. The cat's survival time was approximately 2 years. The other myeloma in a cat that presented with hypercalcemia and renal insufficiency involved visceral organs and produced a biclonal peak due to immunoglobulin A dimer formation on serum electrophoresis. This cat's tumor did not respond to chemotherapy.