Karen V. Jackson

Comparison of five blood-typing methods for the feline AB blood group system

Objective-To compare the ease of use and accuracy of 5 feline AB blood-typing methods: card agglutination (CARD), immunochromatographic cartridge (CHROM), gel-based (GEL), and conventional slide (SLIDE) and tube (TUBE) agglutination assays. Sample Population-490 anticoagulated blood samples from sick and healthy cats submitted to the Transfusion or Clinical Laboratory at the Veterinary Hospital of the University of Pennsylvania. Procedures-Sample selection was purposely biased toward those from anemic, type B, or type AB cats or those with autoagglutination. All blood samples were tested by use of GEL, SLIDE, and TUBE methods. Fifty-eight samples were also tested by use of CARD and CHROM methods. The presence of alloantibodies in all cats expressing the B antigen as detected by use of any method was also assessed. Results-Compared with the historical gold-standard TUBE method, good to excellent agreement was achieved with the other typing tests: CARD, 53 of 58 (91% agreement); CHROM, 55 of 58 (95%); GEL, 487 of 490 (99%); and SLIDE, 482 of 487 (99%; 3 samples were excluded because of autoagglutination). Four of the samples with discordant test results originated from cats with FeLV-related anemia. Conclusions and Clinical Relevance-Current laboratory and in-clinic methods provide simple and accurate typing for the feline AB blood group system with few discrepancies. Retyping after in-clinic typing with the GEL or TUBE laboratory methods is recommended to confirm any type B or AB cats.

Comparison of gel column, card, and cartridge techniques for dog erythrocyte antigen 1.1 blood typing

OBJECTIVE To compare accuracy and ease of use of a card agglutination assay, an immunochromatographic cartridge method, and a gel-based method for canine blood typing. SAMPLE Blood samples from 52 healthy blood donor dogs, 10 dogs with immune-mediated hemolytic anemia (IMHA), and 29 dogs with other diseases. PROCEDURES Blood samples were tested in accordance with manufacturer guidelines. Samples with low PCVs were created by the addition of autologous plasma to separately assess the effects of anemia on test results. RESULTS Compared with a composite reference standard of agreement between 2 methods, the gel-based method was found to be 100% accurate. The card agglutination assay was 89% to 91% accurate, depending on test interpretation, and the immunochromatographic cartridge method was 93% accurate but 100% specific. Errors were observed more frequently in samples from diseased dogs, particularly those with IMHA. In the presence of persistent autoagglutination, dog erythrocyte antigen (DEA) 1.1 typing was not possible, except with the immunochromatographic cartridge method. CONCLUSIONS AND CLINICAL RELEVANCE The card agglutination assay and immunochromatographic cartridge method, performed by trained personnel, were suitable for in-clinic emergency DEA 1.1 blood typing. There may be errors, particularly for samples from dogs with IMHA, and the immunochromatographic cartridge method may have an advantage of allowing typing of samples with persistent autoagglutination. The laboratory gel-based method would be preferred for routine DEA 1.1 typing of donors and patients if it is available and time permits. Current DEA 1.1 typing techniques appear to be appropriately standardized and easy to use.

Evaluation of equine peripheral blood apheresis product, bone marrow, and adipose tissue as sources of mesenchymal stem cells and their differentation potential

OBJECTIVE To evaluate effects of apheresis on mesenchymal stem cells (MSCs) and compare those MSCs with MSCs obtained from adipose tissue or bone marrow (BM). SAMPLE POPULATION Samples obtained from 6 adult horses. PROCEDURES Samples of blood from a peripheral vein, adipose tissue, and BM aspirate were obtained from each horse. Samples were processed via apheresis of blood and techniques reported elsewhere for adipose tissue and BM. Cultures were maintained until adherence and subsequently were subjected to differentiation protocols to evaluate adipogenic, osteoblastogenic, and chondrogenic potential. RESULTS Apheresis product had a significantly higher mononuclear percentage, higher platelet count, and lower RBC count, compared with values for peripheral blood. No cell adherence to the tissue culture plates was detected for the apheresis product. Adherence was detected for 6 of 6 adipose-derived and 4 of 6 BM-derived samples. Variations in efficiency were detected for differentiation of adipose- and BM-derived cells into adipocytes, chondrocytes, and osteoblasts. CONCLUSIONS AND CLINICAL RELEVANCE Apheresis was able to concentrate mononuclear cells and reduce RBC contamination. However, the apheresis product was unable to adhere to the tissue culture plates. In matched horses, adipose- and BM-derived MSCs were capable of producing lipids, glycosaminoglycan, and mineral. The BM was vastly superior to adipose tissue as a source of MSCs with osteoblastogenic potential in matched horses. Additional studies will be necessary to optimize apheresis techniques for horses before peripheral blood can be considered a suitable source for multipotential cells for use in cell-based treatments.

Lymphoma in cats treated with a weekly cyclophosphamide-, vincristine-, and prednisone- based protocol: 114 cases (1998-2008)

OBJECTIVE To evaluate the clinical response rate, progression-free survival time, overall survival time, and possible prognostic factors associated with a cyclophosphamide-, vincristine-, and prednisone (COP)-based chemotherapy protocol in cats with lymphoma. DESIGN Retrospective case series. ANIMALS 114 cats with lymphoma. PROCEDURES Medical records of cats receiving a weekly COP-based chemotherapy protocol from 1998 to 2008 at the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania were evaluated for information regarding signalment, anatomic site of involvement, cell morphology, treatment, and outcome. Retroviral status, baseline weight, substage, anatomic location, dose delays, dose reductions, and response to treatment were evaluated for prognostic importance. RESULTS The majority of cases (94 [82.4%]) were substage b, and the most common anatomic site was the gastrointestinal tract (57 [50%]). Clinical response rate after the first chemotherapy cycle was 47.4%. Response to treatment was significantly associated with progression-free survival time and overall survival time, whereas substage was significantly associated with progression-free survival time. The median progression-free survival time and overall survival time were 65.5 and 108 days, respectively. Compared with nonresponders, responders had significantly longer median progression-free survival time (364 vs 31 days) and median overall survival time (591 vs 73 days). CONCLUSIONS AND CLINICAL RELEVANCE Clinical response after 1 cycle of COP-based chemotherapy was predictive for progression-free survival time and overall survival time in cats with lymphoma; therefore, response after 1 cycle of chemotherapy could be used to guide decisions about further treatment. No new prognostic factors were identified.

Effect of sample storage on blood crossmatching in horses

BACKGROUND Blood samples banked for up to 1 month are typically used to perform pretransfusion testing in humans and small animals, but this has not been validated using blood from horses. HYPOTHESIS Compatibility of equine blood samples is repeatable using fresh samples, and reproducible using donor blood samples stored for up to 4 weeks. ANIMALS Six healthy adult horses. METHODS Randomized, blinded experimental study. Immunologic compatibility of the blood of all horses was assessed using a major and minor saline agglutination and hemolysin crossmatch using blood samples refrigerated for 0-4 weeks and fresh blood from the same horses. Crossmatch results were scored and then compared to identify changes of compatibility in each of the 4 tests. In addition, repeatability of the crossmatch technique itself was assessed by performing 6 iterations of this procedure in immediate succession with fresh blood from 3 horses. RESULTS No significant difference in crossmatch results was found using fresh blood (P = .39-1.00). Reproducibility was poor using blood stored for 1-4 weeks, especially in tests using stored erythrocytes (major antigen crossmatches), with significant differences from baseline at all weeks (P < .05); 13 of these differences were positive, indicating poorer compatibility. CONCLUSIONS AND CLINICAL IMPORTANCE Equine blood crossmatching is repeatable using fresh blood, although decreased apparent compatibility after storage makes exclusion of compatible donors more likely than mistaken administration of incompatible blood. These data suggest that fresh samples should be collected from potential donors before crossmatching equine blood.

Effect of clenbuterol on tracheal mucociliary transport in horses undergoing simulated long-distance transportation

BACKGROUND Pneumonia is observed in horses after long-distance transportation in association with confinement of head position leading to reduction in tracheal mucociliary clearance rate (TMCR). HYPOTHESIS/OBJECTIVES Clenbuterol, a beta-2 agonist shown to increase TMCR in the horse, will ameliorate the effects of a fixed elevated head position on large airway contamination and inflammation in a model of long-distance transportation model. ANIMALS Six adult horses. METHODS A cross-over designed prospective study. Horses were maintained with a fixed elevated head position for 48 hours to simulate long-distance transport, and treated with clenbuterol (0.8 μg/kg PO q12h) or a placebo starting 12 hours before simulated transportation. TMCR was measured using a charcoal clearance technique. Data were collected at baseline and 48 hours, and included TMCR, tracheal wash cytology and quantitative culture, rectal temperature, CBC, fibrinogen, and serum TNFα, IL-10, and IL-2 levels. There was a 18-21 day washout between study arms, and data were analyzed using regression analysis and Wilcoxon rank-sum tests. RESULTS Tracheal mucociliary clearance rate was significantly decreased after transportation in both treatment (P = .002) and placebo (P = .03) groups. There was a significant effect of treatment on TMCR, with the treatment group showing half the reduction in TMCR compared with the placebo group (P = .002). Other significant differences between before- and after-transportation samples occurred for serum fibrinogen, peripheral eosinophil count, quantitative culture, tracheal bacteria, and degenerate neutrophils, though no treatment effect was found. CONCLUSIONS AND CLINICAL IMPORTANCE Treatment with clenbuterol modestly attenuates the deleterious effects of this long-distance transportation model on tracheal mucociliary clearance.

Lycoperdonosis in Two Dogs

Lycoperdonosis is a rare respiratory disease that results from the inhalation of spores released from the Lycoperdon (puffball) mushroom. In the present study, 2 cases of confirmed canine lycoperdonosis are described. The first case presented to the Matthew J. Ryan Veterinary Hospital of the University of Pennsylvania, and the second case was submitted for postmortem examination to the University of Tennessee Veterinary Teaching Hospital. Both dogs presented in respiratory distress, and owners reported that the dogs had been playing or digging in areas with puffball mushrooms prior to the onset of clinical signs. In the initial case, thoracic radiographs revealed a diffuse interstitial and multifocal alveolar pulmonary pattern. Despite aggressive medical treatment and mechanical ventilation, the dog continued to worsen and was euthanized. Postmortem examination revealed firm lung lobes and enlarged tracheobronchial lymph nodes. Histologically, there was a severe diffuse histiocytic and pyogranulomatous bronchointerstitial pneumonia. Throughout the lung and lymph nodes, most commonly within macrophages, were round, 3–5μm in diameter, Gomori methenamine silver—positive structures, consistent with Lycoperdon spores. An approximately 750–base pair DNA fragment was amplified from lung of both cases by polymerase chain reaction using primers specific to yeast ribosomal DNA, and the sequence of the fragment was determined to be most closely related to Lycoperdon pyriforme. Importantly, reexamination of an endotracheal wash from the initial case revealed intrahistiocytic spores, suggesti