Isotype-specific antibodies in horses and dogs with immune-mediated hemolytic anemia

Classes of antibody bound to erythrocytes were determined using direct immunofluorescence (DIF) flow cytometry in 3 horses and 12 dogs with immune-mediated hemolytic anemia (IMHA). Background levels of antibody binding were determined in samples from 12 horses and 12 dogs that were free of clinical disease. The range of nonspecific binding of a fluorescein isothiocyanate (FITC)-conjugated goat anti-equine immunoglobulin G (IgG) was 19.9–36.7%, but was eliminated by the use of the F(ab)₂ fragment of FITC-conjugated goat anti-equine IgG. Background binding by other class-specific antibodies to equine and canine erythrocytes was negligible. The DIF results were compared to the direct antiglobulin (Coombs) test in 5 horses and 20 dogs with anemia. The former assay was more sensitive in dogs with IMHA than was the Coombs' test (100% versus 58%). In contrast, the Coombs' test had better specificity than the DIF assay (100% versus 87.5%, respectively). Using clinical parameters or response to therapy as the comparison, the positive and negative predictive values for the DIF test were 92% and 100% compared to the values of the Coombs' test of 100% and 62%. The DIF assay detected low levels of cells bound with antibody (<30%) in 5 dogs that were Coombs' test-negative. For both species, performance of the DIF test was independent of the prozone effect. Five dogs with IMHA had IgG and IgM on erythrocytes, 5 had IgG, and 2 had IgM. Three horses had surface-bound IgG, including a horse with suspected penicillin-induced IMHA, a foal with neonatal isoerythrolysis, and a foal with clostridial septicemia. The DIF method was valuable in monitoring the response to therapy in the foal with neonatal isoerythrolysis.     

Pharmacokinetics and safety of firocoxib after oral administration of repeated consecutive doses to neonatal foals

The purpose of this study was to determine the pharmacokinetics and safety profile of firocoxib in neonatal foals. Seven healthy foals were administered 0.1 mg/kg firocoxib orally q24 h for nine consecutive days, commencing at 36 h of age. Blood was collected for firocoxib analysis using high‐pressure liquid chromatography with fluorescence detection at 0 (dose #1 only), 0.25, 0.5, 1, 2, 4, 8, 16, and 24 h after doses 1, 5, and 9. For all other doses (2, 3, 4, 6, 7, and 8), blood was collected immediately prior to the next dose (24 h trough). Elimination samples (36, 48, 72, 96, 120, and 144 h) were collected after dose 9. Safety was assessed via physical examinations, body weight measurements, gastroscopy, complete blood count, plasma biochemistry and urinalysis. Firocoxib was rapidly absorbed following oral administration with minimal accumulation after repeat dosing. After the final dose, the terminal half‐life was approximately 11 h. Firocoxib was below the limit of detection (<2.5 ng/mL) in plasma 72 h after the final dose. No significant abnormalities were found on blood analyses, urinalysis, or gastroscopy. This study demonstrated that firocoxib is absorbed in neonatal foals with no demonstrable adverse effects after repeated doses of 0.1 mg/kg.     

Ionophore taste preferences of dairy heifers

Two taste preference studies were conducted using six Holstein heifers in each experiment to determine preferences for no ionophore, lasalocid, or monensin in the diet. In Exp. 1, individually penned (approx. 5 mo old; 220 +/- 14 kg BW) heifers were fed a basal total mixed ration containing 46% corn silage, 46% grass haylage, and 8% soybean meal (DM basis). There were five treatments (mg/kg BW(-1)*d(-1)): 0 ionophore (control), 1 lasalocid (1L), 2 lasalocid (2L), 1 monensin (1M), or 2 monensin (2M). Ionophores were provided as part of the mineral mix that had been added to the control diet and through an ionophore-grain by-product mix to make the 2L and 2M treatments. All five diets were offered for 7 d, with the first 2 d for adaptation and the last 5 d for measurement of feed intake. The most preferred diet was then removed and the study continued with the four remaining diets. The most preferred diets were again eliminated sequentially, so that only two diets remained on d 13 and 14. Each feeding segment ranking of treatment preferences was determined based on the weight of feed refused at the end of each feeding segment. In Exp. 2, six 6-wk-old heifers (75 +/- 5 kg of BW) were individually fed either 0, 1L, or 1M in a study similar to Exp. 1, except that the most preferred diet was removed after 4 d, with the first day for adaptation and the last 3 d for measurement of feed intake. In Exp. 1, orthogonal contrasts indicated that heifers preferred the 1L and 2L diets over the 1M and 2M diets. Preferences between diet concentrations of ionophores (1 and 2 mg/kg of BW; Exp. 1) and the control and ionophore treatments did not differ, nor was there an interaction between ionophores and their concentration. Dairy heifers previously fed lasalocid prefer lasalocid over monensin when given a choice; however, heifers without previous exposure to an ionophore did not indicate a preference (Exp. 2).     

Early growth and environmental implications of dietary zinc and copper concentrations and sources of broiler chicks

1. Environmental accumulation of zinc and copper is becoming a concern in areas having intensive animal production. This study examined performance and excretion of broiler chicks given diets supplemented with graded concentrations of zinc and copper from three different sources. 2. Two experiments were conducted, each utilising 570 1-d-old chicks. In Experiment 1, chicks were given diets containing supplemental zinc concentrations from 40 to 120 mg/kg supplied as zinc sulphate, a zinc amino acid complex (Availa Zn), or a combination of zinc sulphate and Availa Zn with each contributing one-half of the total supplemental zinc. In Experiment 2, broiler chicks were given diets with graded concentrations of supplemental copper ranging from 4 to 12 mg/kg from copper sulphate, a copper amino acid complex (Availa Cu), or a combination of copper sulphate and Availa Cu with each contributing one-half of the total supplemental copper. 3. During the 17-d experimental period, mineral concentration or source did not influence body weight, feed conversion, or the incidence of mortality from 1 to 17 d. 4. Decreasing dietary zinc concentration from 120 to 40 mg/kg reduced zinc excretion by 50%. 5. Copper excretion was decreased by 35% as supplemental copper was reduced from 12 to 4 mg/kg. 6. Reducing dietary zinc and copper concentrations can potentially decrease the accumulation of heavy metals in the environment without compromising bird performance.     

A Hierarchical Perspective to Woody Plant Encroachment for Conservation of Prairie-Chickens

Encroachment of Great Plains grasslands by fire-sensitive woody plants is a large-scale, regional process that fragments grassland landscapes. Using prairie grouse (Tympanuchus spp.) of conservation concern, we apply hierarchy theory to demonstrate how regional processes constrain lower-level processes and reduce the success of local management. For example, fire and grazing management may be locally important to conservation, but the application of fire and grazing disturbances rarely cause irreversible fragmentation of grasslands in the Great Plains. These disturbance processes cause short-term alterations in vegetation conditions that can be positive or negative, but from a long-term perspective fire maintains large tracts of continuous rangelands by limiting woody plant encroachment. Conservation efforts for prairie grouse should be focused on landscape processes that contribute to landscape fragmentation, such as increased dominance of trees or conversion to other land uses. In fact, reliance on local management (e.g., maintaining vegetation structure) to alter prairie grouse vital rates is less important to grouse population persistence given contemporary landscape level changes. Changing grass height, litter depth, or increasing the cover of forbs may impact a few remaining prairie-chickens, but it will not create useable space at a scale relevant to the historic conditions that existed before land conversion and fire suppression.     

Efficacy of Omeprazole versus High‐Dose Famotidine for Prevention of Exercise‐Induced Gastritis in Racing Alaskan Sled Dogs

Background: Omeprazole and famotidine both reduce severity of exercise‐induced gastritis, but administering famotidine is easier than administering omeprazole during racing competition. Hypothesis: Famotidine is more efficacious than no treatment in reducing severity of exercise‐induced gastritis; and high‐dose famotidine is more efficacious than omeprazole in reducing severity of exercise‐induced gastritis. Animals: Experiment 1: Randomized placebo‐controlled study, 36 sled dogs (3–8 years); Experiment 2: Randomized positive‐control study, 52 sled dogs (2–8 years). Methods: Experiment 1: Equal numbers of dogs randomly assigned to famotidine (20 mg q24h) or no treatment groups. Gastroscopy was performed 24 hours after the dogs ran 330 miles. Mucosal appearance was blindly scored by previously described scoring system. Experiment 2: Equal numbers of dogs randomly assigned to omeprazole (20 mg q24h) or high‐dose famotidine (40 mg q12h) groups. Gastroscopy was performed 48 hours before and 24 hours after the dogs ran 300 miles. Mucosal appearance was blindly scored by previously described scoring system. Results: Famotidine reduced the prevalence of clinically relevant, exercise‐induced gastric lesions compared with no treatment (7/16 versus 11/16, P= .031). Compared with high‐dose famotidine, omeprazole significantly decreased the severity (0.4 versus 1.2, P= .0002) and prevalence (2/23 versus 7/21, P= .049) of gastric lesions. Conclusions and Clinical Relevance: Although famotidine provides some benefit in the prevention of exercise‐induced gastric lesions, omeprazole is superior to famotidine in preventing gastritis in dogs running 300 miles. Routine administration of omeprazole is recommended to prevent stress‐associated gastric disease in exercising and racing Alaskan sled dogs.     

Cytology and the cell block method in diagnostic characterization of canine lymphadenopathy and in the immunophenotyping of nodal lymphoma

Minimally invasive techniques used to evaluate canine peripheral lymphadenopathy (PLN), including fine needle aspiration biopsy with cytological evaluation (FNAB‐C) and flow cytometry (FC), have benefits and limitations. The cell block (CB) method is an alternate processing technique in which fine needle aspirate biopsy samples are concentrated, fixed, and embedded in paraffin for routine histological processing/staining. Utilizing three observers, we determined the diagnostic value of the CB in evaluating canine PLN across six categories (non‐diagnostic, reactive, inflammatory/infectious, probable lymphoma and lymphoma, metastatic neoplasia) and correlated findings to immunophenotypic and clonal antigen receptor rearrangement results in canine nodal lymphoma. Eighty‐five paired FNAB‐C and CB samples were evaluated from canine patients presenting to the University of Minnesota Veterinary Oncology or Internal Medicine services. Diagnostic quality samples were obtained in 55/85 (65%) CB and 81/85 (95%) FNAB‐C samples, respectively, and nodal pathology impacted CB diagnostic yield. Overall percent agreement between diagnostic‐quality FNAB‐C and CB samples was 86%, but increased to 95% if the categories of lymphoma and probable lymphoma were combined. There was 100% agreement for both the diagnoses of metastatic neoplasia and reactive lymph nodes and 92% agreement for the diagnosis of lymphoma/probable lymphoma. Using immunohistochemistry (IHC), CB samples correctly immunophenotyped 22/23 (96%) cases of B‐cell lymphoma, but only 1/6 (17%) cases of T‐cell lymphoma. IHC was not completed on nine cases of lymphoproliferative disease because of insufficient cellularity. When the CB method (CBM) yielded diagnostic quality samples there was good to excellent agreement with FNAB‐C samples and CB samples were suitable for some IHC tests.     

Somatic cell mapping and restriction fragment length polymorphism analysis of bovine insulin-like growth factor I.

The DNA isolated from cow-hamster hybrid somatic cells segregating bovine chromosomes was analyzed by Southern blotting and hybridization with a heterologous [32P]-labeled porcine cDNA probe encoding insulin-like growth factor I (IGF-I). Thirteen of 25 cow-hamster hybrid cell lines exhibited the bovine-specific IGF-I fragment. Analysis for the retention or loss of bovine IGF-I with markers previously screened against the same panel of hybrid cells revealed a 100% concordance with lactate dehydrogenase B of bovine syntenic group U3 located on bovine chromosome 5. Restriction fragment length analyses of genomic DNA from animals representing five breeds (Angus, Polled Hereford, Simmental, Gelbvieh, and Belgian Blue) and from seven half-sib Angus calves indicated that polymorphisms for the genomic composition of the bovine IGF-I gene may exist in cattle populations.