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Summary

A simple cryogenic technique for preserving bovine buffy coat leukocytes was developed. This was coupled with a variation of the standard discontinuous gradient technique to purify mononuclear cells that retained immunologic function. The total number of mononuclear cells recovered from cyropreserved samples were only 87 to 42% of those recovered from freshly obtained blood samples. However, the functional capabilities of mononuclear cells from cyopreserved buffy coat preparations were retained. Polyclonal proliferative responses to 3 mitogens were measured, using a titration of mitogen concentrations, and were found to be normal, compared with those of cells isolated from fresh blood. Blood samples collected after vaccination with Brucella abortus contained leukocytes that responded to irradiated B abortus. These antigen-specific responses were also retained through cyopreservation. Cell surface expression of T-lymphocyte antigens, CD2, CD4, and CD8, and cell-surface IgM on B lymphocytes was also evaluated. Flow cytometric analysis of fresh and cryopreserved mononuclear cell preparations indicated that the relative proportions of different subpopulations were not altered. The technical simplicity of our cryopreservation system will allow processing of large numbers of samples with the ability to assay various immune functions at a later time.

Free access
in American Journal of Veterinary Research

Abstract

Objective—To determine prevalence of anthelmintic resistance in cyathostome nematodes of horses in the southern United States.

Design—Cross-sectional study.

Animals—786 horses on 44 farms and stables in Georgia, South Carolina, Florida, Kentucky, and Louisiana.

Procedure—Fecal egg count (FEC) reduction tests were performed on 44 large farms and stables. Horses on each farm were treated with an oral paste formulation of fenbendazole, oxibendazole, pyrantel pamoate, or ivermectin at recommended label dosages. A mixed linear model was fitted to the percentage reduction in FEC, accounting for differences among farms, states, ages, treatments, and treatment by state interactions.

Results—By use of a conservative measure of resistance (< 80% reduction), the percentage of farms with anthelmintic-resistant cyathostomes was 97.7%, 0%, 53.5%, and 40.5% for fenbendazole, ivermectin, oxibendazole, and pyrantel pamoate, respectively. Mean percentage reductions in FEC for all farms were 24.8%, 99.9%, 73.8%, and 78.6% for fenbendazole, ivermectin, oxibendazole, and pyrantel pamoate, respectively. Pairwise contrasts between states for each treatment revealed that in almost all instances, there were no significant differences in results between states.

Conclusions and Clinical Relevance—The prevalence of resistance found in this study was higher than that reported previously, suggesting that anthelmintic resistance in equine cyathostomes is becoming a major problem. Furthermore, data from these 5 southern states, which are geographically and physiographically distinct, were remarkably similar. This suggests that drug resistance in cyathostomes is highly prevalent throughout the entire southern United States and probably nationwide. (J Am Vet Med Assoc 2004;225:903–910)

Full access
in Journal of the American Veterinary Medical Association

Abstract

Jejunal hemorrhage syndrome (JHS) is an acute, highly fatal enterotoxemic disorder in dairy cattle that has been reported during the last few decades. No specific cause of this syndrome has been identified; however, several studies have revealed a strong association between JHS and infection with Clostridium perfringens type A. A common mold, Aspergillus fumigatus, has also been implicated as a potential causative agent in this disease syndrome. Clinical signs of JHS (including sudden decreases in feed intake and milk production, rapid loss of condition, a right-sided ping audible during simultaneous auscultation and percussion of the abdomen, abdominal distension, and melena or bloody feces) usually develop early during lactation when cattle receive rations that are high in energy and low in fiber. Appropriate preventive strategies have not yet been determined, and intensive medical management with or without surgical intervention is rarely successful. The use of commercially available vaccines that are directed against C perfringens types C and D is of questionable efficacy and not likely to be helpful as a preventative measure. This article highlights the potential etiologic and risk factors, describes common clinical signs, outlines relevant diagnostic testing, and summarizes treatment options and their outcomes.

Full access
in Journal of the American Veterinary Medical Association