Citrate inhibition of snake venom proteases.

Thirty snake venoms had a citrate content of 2.3 to 12.9%, dry basis, by an aconitase isocitric dehydrogenase coupled enzyme assay. This is a venom concentration range of approximately 30 to 150 mM citrate assuming 25% venom solids content. Inhibition of snake venom protease activity by the addition of exogenous citrate was obtained using azure blue hide powder and azocasein as substrates. Protease inhibitions of 7.5% for Crotalus atrox venom to 78% for Bothrops picadoi venom were observed with citrate. Complete inhibition of snake venom protease activity by citrate was not observed. Bothrops asper (Pacifico) venom showed a 41% protease inhibition by citrate with azocasein as the substrate and 46% inhibition of Bothrops asper (Alantico) venom protease with azure blue hide power as a substrate. Trypsin was not inhibited in this system. Citrate may inhibit some venom protease activity by forming a complex with the zinc of zinc-dependent enzymes. reserved.

Failure to transmit Ehrlichia canis (Rickettsiales: Ehrlichieae) with Otobius megnini (Acari: Argasidae).

An ear tick, Otobius megnini (Dugès) recovered from a child who had serologic evidence of ehrlichiosis, was examined for Ehrlichia species microscopically and by inoculation into a susceptible dog; no evidence of infection was found in the tick. Experimental transmission of E. canis by laboratory-reared O. megnini was attempted; neither transstadial nor transovarial transmission occurred.

The acute and subchronic toxicity of BRB-I-28, a novel class Ib antiarrhythmic agent, in CD-1 mice.

The acute and subchronic toxic effects of BRB-I-28 (7-benzyl-3-thia-7-azabicyclo[3.3.1]nonane HCl), a novel class Ib antiarrhythmic agent, were investigated in male and female mice. The estimated oral LD(50) for BRB-I-28 was 128 mg/kg (male mice) and 131 mg/kg (female mice). In subchronic oral studies, four groups of mice (15/sex/group/dose) were fed daily with diets containing BRB-I-28 for 90 consecutive days. The equivalent daily doses were approximately 0, 16, 32, 76 (male) and 0, 18, 37, 89 mg/kg (female). All mice survived. Food consumption per day was decreased, but water consumption per day was increased (in a non-dose-dependent manner). However, both mean body weight and mean body weight gain were not significantly changed as were true for hematological and clinical chemistry profiles, except for serum Na(+) concentration (male) and serum K(+) concentration in male and female mice (high dose levels). Hepatocellular necrosis occurred in male and female mice (in a dose-dependent fashion). Renal cortical vacuoles and myocardial necrosis with low numbers of lymphocytic infiltrations were present in female mice (middle and high doses). Lesions in the liver, kidney and heart were mild with (very small) changes in serum biochemical values. These data suggest that BRB-I-28 has limited toxic potential, and coupled with low proarrhythmic and other desirable cardiovascular effects, makes BRB-I-28 worthy of further development.

Detection of anticytauxzoon antibodies in cats infected with a Cytauxzoon organism from bobcats.

A microfluorometric immunoassay was developed for the detection of antibodies to a Cytauxzoon organism of bobcat origin. A homogenated antigen of RBC-parasite lysate was used to measure antibody levels in sera from cats experimentally infected with the non-pathogenic erythrocyte form of the bobcat Cytauxzoon organism. Antibody levels in sera collected at weekly intervals, expressed as FIAX values, were correlated with parasitemia levels and erythrocyte packed cell volume (PCV). The antibody levels tended to be highest and the PCV lowest at or near the peak parasitemias. Splenectomized cats developed higher antibody and parasitemia levels than non-splenectomized cats. Single serum samples collected from 31 cats were used to establish a baseline response level for normal domestic cat populations.

An overview of serological tests currently available for laboratory diagnosis of parasitic infections.

Current methods and commercial test systems for the diagnosis of parasitic infections in both animals and humans are reviewed. Lists of test kits and their manufacturers are provided along with ordering information: the only commercially available test kits are for the diagnosis of toxoplasmosis in humans or animals and dirofilariasis (heartworm) in dogs. A partial list of diagnostic laboratories and the parasite tests they perform is also provided. Complete lists of diagnostic tests that could be obtained in the private sector are not available but would be useful. Two microfluorometric solid-phase assay systems are reviewed, and adaptations to custom assays for several kinds of parasites are briefly described. User problems in performing tests and interpreting results are stressed with emphasis placed on diagnosis of dirofilariasis in dogs. False-positive serology in dogs without heartworms and negative antibody responses in micro-filariae-positive animals are discussed with respect to proper interpretation of results.

X-ray powder diffraction and microscopic analysis of crystalluria in dogs with ethylene glycol poisoning.

Most of the urinary crystals in 2 dogs intoxicated with ethylene glycol had prismatic (rod-like, hippurate-like), and to a less extent, hemp seed or spindle shapes. Using X-ray powder diffraction and optical analysis, these crystals were identified as calcium oxalate monohydrate. A few crystals of calcium oxalate dihydrate (envelope shape) were also observed in the urine sediment. Their presence was confirmed by the appearance of weak X-ray peaks having interplanar spacings ranging from 0.617 to 0.620 nm, which are thought to correspond to the 200 X-ray reflection of calcium oxalate dihydrate. Thus, the 6-sided hippurate-like crystals in the urine of ethylene glycol-poisoned dogs are actually calcium oxalate monohydrate.

Familial canine dermatomyositis: clinicopathologic, immunologic, and serologic studies.

Laboratory studies were performed on 3 Collies with familial canine dermatomyositis, 6 progeny from a breeding of 2 of the Collies (incross litter), and 4 progeny from the breeding of an affected Collie male and a normal Labrador Retriever (outcross litter). Hematologic abnormalities were leukogram changes consistent with inflammation, and, in 2 severely affected incross dogs, anemia of inflammatory disease. Serum muscle enzyme activities were not markedly increased. High concentrations of immune complexes and proportionally increased total immunoglobulin G were present in the sera of moderately and severely affected incross dogs. The same dogs had weakly positive direct Coombs' tests, and 1 affected outcross dog had a strongly positive direct Coombs' test. Rheumatoid factor was present in a severely affected incross dog. A few dogs had polyclonal globulin increases that were attributed to inflammation. Low antibody titers to canine calicivirus were present in 3 dogs and a low titer to canine coronavirus was present in 1 dog.

Survival of a domestic cat with naturally acquired cytauxzoonosis.

A cat with acute onset of febrile systemic illness was determined to be infected with Cytauxzoon felis. The diagnosis was made on the basis of cytologic morphology of erythroparasites seen in blood smears and results of microfluorometric immunoassay for serum antibody directed against C felis-parasitized RBC. Treatment consisted of parenteral administration of fluids and antibiotics. The cat recovered within 2 weeks. Circulating erythroparasites were not detected on blood smears from samples collected during follow-up examinations. However, high serum antibody titer persisted for at least 15 weeks after infection. The cat continued to be free of clinical disease 2.5 years after the initial diagnosis. Whether C felis infection persists in this cat has yet to be determined. This case indicates that some domestic cats can recover from naturally acquired cytauxzoonosis.

Disseminated histoplasmosis in cats: 12 cases (1981-1986).

Anemia, weight loss, lethargy, fever, anorexia, and interstitial lung disease were the predominant clinical findings in 12 cats with disseminated histoplasmosis. Some cats were examined because of dysfunction or lesions of bone, eyes, or skin. In most cases, the clinical signs were observed by the owner for 4 weeks or less before seeking veterinary care. Young cats were most commonly affected, with 7 of the 12 cats less than or equal to 1 year old. Identification of Histoplasma organisms in bone marrow aspirates was used to confirm the diagnosis of histoplasmosis in 11 of the 12 cats. Histoplasma infection of multiple organs was found at necropsy. In this study, disseminated histoplasmosis had a higher prevalence in cats than in dogs at the same veterinary medical teaching hospital. Feline disseminated histoplasmosis was not associated with FeLV infection. Treatment was attempted in 7 of the 12 cats.

Disseminated histoplasmosis in dogs: 12 cases (1981-1986).

Diarrhea, intestinal blood loss, anemia, and lethargy were predominant clinical findings in 12 dogs with disseminated histoplasmosis. Young dogs were affected most commonly, with 6 dogs being 1 to 3 years old. A diagnosis of disseminated histoplasmosis was established on the basis of histologic or cytologic detection of Histoplasma organisms in intestinal or rectal mucosa in 7 dogs, in circulating leukocytes in 5 dogs, in bone marrow in 3 dogs, and in multiple tissues at necropsy in 1 dog (4 dogs had Histoplasma organisms detected in greater than 1 site). Anemia was detected in 10 dogs (PCV less than 20% in 3 dogs), and the anemia was inadequately regenerative or nonregenerative in 7. Hypoalbuminemia was detected in 9 dogs, and serum albumin concentrations were low (less than 1.0 g/dl) in 4 of the 9 dogs. Of 5 dogs treated with ketoconazole, 2 were in remission for greater than or equal to 1 year. Corticosteroid therapy may have exacerbated the disease in 4 dogs. Histoplasma infection of multiple organs was detected in 5 necropsied dogs.

Identification of Histoplasma organisms in circulating eosinophils of a dog.

Disseminated histoplasmosis was diagnosed in a 10-year-old dog that had chronic diarrhea, weight loss, fever, and anemia. The diagnosis was based on detection of Histoplasma organisms in circulating neutrophils, monocytes, and eosinophils. The dog had severe histoplasmal fungemia, which may have been caused by treatment with prednisolone.

Lymphocytic-plasmacytic enteritis in two horses.

The primary hematologic abnormalities in 2 adult horses with chronic weight loss were hypoalbuminemia and hyperglobulinemia. One horse was anemic, had subclinical disseminated intravascular coagulation, and prolonged plasma sulfobromophthalein half-life. Small-intestinal dysfunction with malabsorption was indicated by abnormal D-xylose absorption test results. Clinicopathologic and pathologic findings were consistent with a diagnosis of malabsorption and protein-losing enteropathy, attributable to lymphocytic and plasmacytic infiltration of the intestine.

Severe systemic reactions to Hymenoptera stings in three dogs.

Three dogs were treated for acute severe systemic reactions following Hymenoptera stings. The reactions were characterized clinically by CNS depression, shock, and hemorrhage, and clinicopathologically by inflammation, liver injury, renal disease, hypoproteinemia, and possible disseminated intravascular coagulation. The severe systemic reaction may have resulted from allergic mechanisms, toxic, nonimmunologic mechanisms, or both. Rapid correction of hypovolemia and prevention of vascular stasis are the most important aspects of treatment.

Myxosarcoma in a Sinaloan milksnake.

A 2-year-old Sinaloan milksnake was examined because of a 1-cm mass attached to the lateral wall of the coelom. A diagnosis of myxosarcoma was made on the basis of histologic features and special staining characteristics.

Renal concentrating ability in dehydrated hyponatremic dogs.

Eleven hyponatremic dogs were unable to concentrate their urine during periods of severe dehydration and azotemia. When normonatremia was reestablished in eight of the dogs, their renal concentrating ability returned. Six dogs, including the 3 dogs in which normonatremia was not reestablished, died or were euthanatized; renal lesions were not found during postmortem examination. Two dogs had hypoadrenocorticism, which has been documented as a cause of hyponatremia and impaired renal concentrating ability. Two dogs had gastrointestinal disease, which has been documented as a cause of hyponatremia, but not of impairment of renal concentrating ability. All dogs without hypoadrenocorticism had clinical and clinicopathologic indications of blood loss, which has not been documented as a cause of hyponatremia or impairment of renal concentrating ability. Hyponatremia (less than 120 mEq/L) was induced by chronic blood removal in a dog maintained on a low-sodium diet. During the period of hyponatremia, the dog became azotemic, hypotensive, and severely dehydrated; renal concentrating ability was impaired. We conclude that hyponatremia may be caused by hemorrhage, but irrespective of the cause, hyponatremia impairs renal concentrating ability.

Ehrlichiosis and polyarthritis in three dogs.

Three dogs admitted for evaluation of lameness were determined to be infected with a neutrophilic strain of Ehrlichia. Ehrlichia morulae were detected in low numbers in both synovial fluid and blood neutrophils. The dogs had rapid clinical improvement after appropriate antibiotic therapy. A diagnosis of canine ehrlichiosis easily could have been missed if morulae had not been identified. Ehrlichiosis should be considered in acutely lame dogs residing in areas enzootic for ehrlichiosis. A careful search for Ehrlichia morulae within synovial fluid and blood neutrophils should be performed. An E canis titer determination also may be helpful.

Ehrlichiosis in a dog with seizures and nonregenerative anemia.

Ehrlichia canis infection was diagnosed in a dog with a history of seizures and nonregenerative anemia. Serologic titer to E canis was greater than 1:100. Evaluation of CSF revealed a high cell count, high protein concentration, and a positive Pandy test result. Several mononuclear leukocytes in the CSF contained E canis morulae. Central nervous system lesions are commonly found on postmortem examination of animals with ehrlichiosis, although clinical reports of neurologic signs attributable to this disease are less common. Ehrlichiosis should be included in the differential diagnosis of CNS disease in dogs from enzootic areas.

Pneumocystis carinii pneumonia in foals.

Pneumocystis carinii pneumonia was diagnosed in 3 foals. In 2 foals (No. 1 and 2), diagnosis was by histologic evaluation of pulmonary tissue. On retrospective evaluation, P carinii cysts were found on sediment smears of bronchoalveolar lavage fluid in 1 foal (No. 1). A different foal (No. 3) was diagnosed as having pneumocytosis by finding P carinii cysts in bronchoalveolar lavage fluid, and was treated successfully. Definitive diagnosis of pneumocytosis in animals is usually made at necropsy. However, careful cytologic evaluation in bronchoalveolar lavage fluid sediment can provide a diagnosis in some cases, allowing for initiation of appropriate treatment.

Cytology of cutaneous lesions.

Cytology is a quick, easy, and inexpensive way to gain useful information concerning the process(es) or etiology of cutaneous lesions. Cutaneous lesions can be separated quickly into inflammatory or noninflammatory conditions and organisms and neoplastic cells can be identified, allowing for a definitive diagnosis. Although not all lesions can be diagnosed cytologically, recognition of the process(es) (for example, purulent inflammation, eosinophilic inflammation, cysts) allows for better therapeutic management and a more accurate prognosis.

Evaluation of pleural and peritoneal effusions.

Certain diseases cause an increase in the amount of fluid present in the pleural and/or peritoneal cavity (an effusion). Uroperitoneum subsequent to kidney, ureter, bladder, or urethra rupture also can cause an increased amount of fluid in the abdomen. Evaluation of fluid samples often is helpful in identifying the mechanism causing the effusion and, occasionally, results in a specific diagnosis. The TP, TNCC, and general cytologic examination can be performed easily, quickly, and inexpensively in-house. The TP and TNCC are used to classify effusions as transudates, modified transudates, or exudates. Transudates usually are caused by hypoalbuminemia, but also can be caused by leakage of fluid from efferent intestinal lymphatics. Cytology and culture usually are not rewarding in the evaluation of transudates. Modified transudates usually are caused by increased vascular permeability or increased intrahepatic hydrostatic pressure. Cytologic and radiographic examinations often are helpful in evaluating patients with modified transudates, while cultures usually are unrewarding. The exudate class encompasses the inflammatory exudates (septic or nonseptic), neoplastic exudates, and chylous effusions. Inflammatory exudates have a high TP and predominantly contain inflammatory cells. They may be septic or nonseptic. When septic, degeneration neutrophils often, but not always, are found. Cultures often are needed to determine whether sepsis is present, to identify the specific organism, and to determine the best therapy. Neoplastic exudates may contain numerous neoplastic cells. If there is concern that the cells are dysplastic instead of neoplastic, the cytology preparation should be referred to a consultant. Chylous effusions usually contain many small lymphocytes with a variable number of neutrophils and macrophages. In chronic chylous effusions, however, neutrophils and/or macrophages may predominate. Chylous effusions usually are differentiated easily from pseudochylous effusions by cytology. Comparison of fluid and serum triglyceride and cholesterol concentrations can be used to differentiate chylous and pseudochylous effusions when differentiation cannot be accomplished by cytology.