Flow cytometric evaluation of bound IgG on erythrocytes of anaemic dogs

The detection of red blood cell (RBC)-bound immunoglobulins in case of anaemia with the direct agglutination test (DAT or Coombs test) has been reported to be of low sensitivity. We therefore tested the applicability of flow cytometry for the detection of canine IgG on RBC using two different IgG-specific secondary reagents: goat-anti-dog IgG (GalphaD-IgG) and rabbit-anti-dog IgG (RalphaD-IgG). Membrane staining RBC samples were performed at 4 degrees C. Comparisons of agglutination test at 37 degrees C and 4 degrees C showed, that binding of the secondary antibodies at 4 degrees C was more sensitive compared to agglutination at 37 degrees C and the two antisera differed considerable in their agglutination activity. Binding of GalphaD-IgG and RalphaD-IgG to RBC of healthy dogs (n=15) was low and mean fluorescence intensities were taken to calculate thresholds above which RBC of patients were judged positive. As in agglutination tests, both secondary antisera displayed considerable differences (concentration-dependent binding and histogram profiles) after flow cytometric analysis. Using flow cytometry, with GalphaD-IgG 8 of 17 agglutination-negative patients were positive and RalphaD-IgG was positive with 3 of 3 agglutination-negative RBC samples. Thus, flow cytometric analysis of proved to be a sensitive technique, detecting RBC-bound canine IgG of DAT-negative patients. The results of both techniques, however, are significantly influenced by the used IgG-specific polyclonal reagents.     

Dissipation and transport of clopyralid in soil: effect of application strategies

At present there are no herbicide uses registered for broadleaf weed control in buckwheat. Clopyralid, mixed with desmedipham, was anticipated to provide early-season broadleaf weed suppression with minimal crop injury. However, field trials resulted in limited success, which brought into question the fate and availability of clopyralid for weed control. A 4-year field study was conducted in Lithuania to evaluate the dissipation of clopyralid in Haplic Luvisol sandy loam soil and the influence of application rate, application timing, and coapplication with desmedipham on its degradation and transport. Clopyralid dissipation was rapid; 50% dissipation times, in the surface 5 cm, averaged over the 4 years of the study, were <7 days. Application rate (90 versus 180 g ai ha(-1)), timing (pre-emergence versus postemergence), and coapplication with desmedipham (360 g ai ha(-1)) did not significantly influence clopyralid dissipation. Clopyralid dissipation by leaching was not a significant factor; at 7-21 days after application, <8 μg kg(-1) was found at the 10-20 cm depth. Understanding the dissipation of herbicides and the influence of application strategies on herbicide fate will allow for informed decisions and improved efficacy of weed control. On the basis of the results of this research, weed scientists can now determine whether increased rates of clopyralid would provide sufficient residual chemical for adequate weed control without crop injury.     

Reactivity of cross-reacting monoclonal antibodies with canine leukocytes, platelets and erythrocytes

A panel of 380 commercially available monoclonal antibodies (mAbs) against human CD molecules from various sources was tested during the 8th Human Leukocyte Differentiation Antigen Workshop (HLDA8) for cross-reactivity on canine peripheral blood leukocytes by flow cytometry. In addition, all mAbs were used to label a 50:50 mixture of platelets and erythrocytes of the same dogs. This testing resulted in 51 cross-reacting mAbs. mAbs with specificity for CD9, CD29, CD42a, CD61, and CD41/CD61 showed cross-reactivity with canine platelets in a non-polymorphic and one mAb with the erythrocyte antigen CD235a in a polymorphic reaction pattern. Canine leukocyte-reactive mAbs included those with specificity for CD11a, CD11b, CD14, CD18, CD21, CD22, CD47, CD49d, CD49e, CD56, CD62L, CD91, CD94, and CD172a. In addition, several mAbs resulted in a staining pattern of canine cells which suggest that the canine epitope equivalents have an alternate expression pattern from that expected for humans (CD1a, CD35, CD44, CD45, CD75s, CD81). In summary, this study confirmed the reactivity of previously described cross-reactive mAbs with canine cells and resulted in the characterization of mAbs recognizing so far undetectable canine CD molecules.