Comparison of dot blot hybridization, polymerase chain reaction, and virus isolation for detection of bovine herpesvirus-1 (BHV-1) in artificially infected bovine semen.

Bovine semen samples spiked with bovine herpesvirus 1 (BHV-1) were used to compare dot blot hybridization, polymerase chain reaction (PCR), and virus isolation for detection of BHV-1 in bovine semen. The PCR amplification used primers targeting the BHV-1 thymidine kinase gene and a nucleic acid releasing cocktail (GeneReleaser); the PCR product was used as the DNA probe in dot blot hybridization; virus isolation was done in primary bovine fetal testis (BFT) cell cultures. Semen diluted 1:20 in tissue culture medium had the least cytotoxicity and inhibition of viral cytopathic effects in BFT cells, allowing detection of 1 TCID50/100 microL of BHV-1 suspension by virus isolation. The presence of foreign DNA such as bovine sperm DNA or salmon sperm DNA increased the sensitivity of dot blot hybridization in detecting BHV-1, allowing detection of 20,000 TCID50/100 microL of neat semen. The inhibition of PCR amplification of BHV-1 DNA in bovine semen was eliminated by diluting the samples 1:20 in tissue culture medium. The best PCR amplification was obtained when semen was diluted 1:20 and when a reaction buffer of pH 9.0, with 1.0 mM MgCl2 was used. Under these conditions, the PCR followed by ethidium bromide staining of agarose gels could detect 1 TCID20/100 microL of sample, whereas PCR followed by Southern blot hybridization could detect 0.01 TCID50/100 microL of sample.(ABSTRACT TRUNCATED AT 250 WORDS)     

Temporary Bile Diversion in Cats with Experimental Extrahepatic Bile Duct Obstruction

The use of a cholecystostomy catheter for temporary bile diversion was investigated in four cats with experimentally induced extrahepatic bile duct obstruction. Eighteen days after ligation of the common bile duct, a 6.5 F accordion catheter was placed in the gallbladder with a 22 g Hawkins needle-guide system through a paracostal incision. Biochemical parameters and fasting serum bile acids were monitored for 16 days. There were significant decreases in mean total bilirubin, aspartate aminotransferase, and fasting serum bile acids within 72 hours of bile diversion, and in mean alanine aminotransferase within 96 hours. Attitude and appetite improved, and the catheter was tolerated well. Positive bile cultures developed in three cats. Histologic changes in the gallbladder included mucosal ulcerations, a mixed inflammatory cellular infiltration, and fibrosis of the submucosa.     

Effect of inoculum density of Macrophomina phaseolina on seedling susceptibility of six conifer species

The severity of charcoal root disease of conifer seedlings (Abies concolor, A. magnified, Pinus jeffreyi, P. lambertiana, and Pseudotsuga menziesii) was a power function of the sclerotial density of the fungus (Macrophomina phaseolina) in nursery soil.     

Quantitative Aspects of Thyroid Scintigraphy With Pertechnetate (99mTcO4) in Cats

Thyroidal ^99mTcO₄(pertechnetate) uptake percentages were determined in unanesthetized euthyroid (n = 13) and hyperthyroid (n = 18) cats. Maximal uptakes were observed 60 minutes after IV injection of the radionuclide and ranged from 0.3 to 3.9% of the dose in euthyroid cats (median 2.23%) and from 5.2% to 23.9% of the dose in hyperthyroid cats (median 14.8%) ( P < .05). There were no overlaps in pertechnetate uptake percentages during any of the intervals evaluated. It is concluded that the optimal time for visualization of the thyroid by ^99mTcO₄-scanning is 60 minutes after IV injection of the radionuclide. Calculation of the percentage uptake is of additional diagnostic value.     

Necrotizing typhlocolitis associated with a spirochete in rheas (Rhea americana).

Necrotizing typhlocolitis was diagnosed in 13 juvenile common rheas (Rhea americana) from three separate of geographically isolated Ohio flocks, with mortality ranging from 25% to 80%. At postmortem examination, a diphtheritic membrane covered ulcerated cecal mucosa. Histologically, cecal sections showed necrosis and granulomatous-to-suppurative inflammation that extended into the submucosa and often surrounded large eosinophilic colonies of bacteria. Warthin-Starry staining showed these colonies to be composed of entangled spirochetes that invaded the submucosa and frequently were present transmurally. Similar organisms were identified by Warthin-Starry staining in the cecum of a juvenile rhea from a fourth flock that histologically had mild lymphocytic typhlitis. Scanning and transmission electron microscopy demonstrated the presence of a spirochete in the ceca. Anaerobic culture yielded a gram-negative, beta-hemolytic spirochete. Coccidia, histomonads, and Salmonella spp. were consistently absent.     

Effects of recombinant canine granulocyte colony-stimulating factor on white blood cell production in clinically normal and neutropenic dogs.

Recombinant canine granulocyte colony-stimulating factor (rcG-CSF) was administered to clinically normal dogs, cyclic-hematopoietic dogs, and dogs undergoing autologous bone marrow transplantation, to determine whether rcG-CSF could be used to stimulate WBC production and function in normal and neutropenic dogs. To the normal dogs, rcG-CSF was administered by SC injection at rates of 1 microgram/kg of body weight, q 12 h; 2 micrograms/kg, q 12 h; or 5 micrograms/kg, q 12 h. A significant dose-dependent increase in the WBC count resulted from the stimulation of bone marrow progenitor cells. The increased WBC count was characterized by mature neutrophilia and monocytosis. Neutrophil myeloperoxidase and phagocytic activity were normal in rcG-CSF-treated normal dogs, demonstrating the production of normal functional neutrophils in response to rcG-CSF treatment. Recombinant canine G-CSF prevented neutropenia and associated clinical signs but did not completely eliminate the cycling of neutrophils in cyclic-hematopoietic dogs when it was administered at rates of 1 microgram/kg, q 12 h, and 2.5 micrograms/kg, q 12 h. The time to bone marrow reconstitution was not decreased in dogs treated with rcG-CSF at a rate of 2.5 micrograms/kg, q 12 h, for 13 days following autologous bone marrow transplantation. On the basis of our findings, we suggest that treatment with rcG-CSF is an effective way to stimulate myelopoiesis in dogs, but that the dose of rcG-CSF required to stimulate WBC production will vary depending on the cause of neutropenia. Recombinant canine G-CSF should be useful in stimulating production and maintaining function of WBC for treatment of clinical diseases seen commonly in veterinary practice.     

Coccidioidomycosis in 48 Cats: A Retrospective Study (1984–1993)

Coccidioidomycosis was diagnosed in 48 cats. Forty-one cases were identified within a period of 3 years. Coccidioides immitis was revealed by cytological or histopathological examinations, or culture in 70% of cats. The remaining 30% of cases were diagnosed by appropriate clinical signs, radiographic lesions, and serological test results. The average age of affected cats was 6.2 years with a median age of 5.0 years. Fifty-four percent (n = 26) were female and 46% (n = 22) were male. Domestic shorthaired and longhaired breeds comprised 89% (n = 41) of affected cats. Sixty-seven percent of cases were diagnosed during the 6-month period of December through May. Cats infected with C immitis were presented for evaluation of dermatologic (56%), respiratory (25%), musculoskeletal (19%), and neurological or ophthalmologic signs (19%). Fever, inappetence, and weight loss were present in 44% of the cats. Duration of clinical signs before diagnosis was less than 4 weeks in 85% (n = 42) of cats, with an average of 3.8 weeks and a median of 2 weeks. Agar gel immunodiffusion tests were positive in all 39 cats tested at sometime during the course of their disease. Hyperproteinemia (greater than 7.9 g/dL) was present in 52% (10/23) of cases. The majority of cats (n = 39) were negative for feline leukemia virus. Antibodies to feline immunodeficiency virus were absent in the 19 cats tested. Ketoconazole was the most common antifungal agent used to treat cats with Coccidioidomycosis. Duration of treatment ranged from less than 1 week to 43 months. Thirty-two cats are currently asymptomatic, with or without treatment. Eleven cats died or were euthanized. Five cats were lost to follow-up. Ketoconazole likely is more suppressive than curative because relapses were common after discontinuing therapy.     

Effect of age on absorption and immune responses to weaning or introduction of novel dietary antigens in pigs

Pigs weaned at three weeks old absorb food protein antigens from the intestine. The amount of antigen absorbed declines over the next three weeks, and this decline is associated with an increasing level of serum antibody to the fed proteins. There was no difference in the rate of immune elimination of intravenously injected antigen in fed and unfed controls. The reduction of serum antigen is thus likely to reflect reduced absorption, possibly mediated by locally produced antibody. Pigs weaned at 10 weeks old also absorbed antigens and produced an antibody response when introduced to soya; and after three weeks of feeding soya the absorption of antigen was substantially reduced. This latter exclusion was specific for soya as a second novel protein (ovalbumin) was absorbed when introduced to the diet at this time. At six months, pigs no longer absorbed soya proteins when they were introduced to the diet. Furthermore, pigs of this age had serum 'antibody' to soya and other proteins such as keyhole limpet haemocyanin to which they had never been exposed.