RESUMEN
The immunodiagnostic assessment of dogs suspected of having immune-mediated hemolytic anemia (IMHA) is based on persistent autoagglutination of erythrocytes (after three saline washes), marked spherocytosis, and a positive direct antiglobulin (Coombs') test (DAT). However, the value of using the indirect antiglobulin test (IAT) for the detection of anti-erythrocytic autoantibodies in serum from dogs suspected of having IMHA is unclear. To evaluate the IAT, leftover serum samples from a large cohort of 94 dogs suspected of having IMHA and for which DAT results were known were incubated with DAT- erythrocytes, and five IAT techniques were performed (in part with different reagents and temperatures): microtiter plate (MICRO), microcapillary, laboratory gel column, gel minitube kit (GEL KIT), and immunochromatographic strip kit. Two IAT techniques (MICRO at 37 °C and GEL KIT with rabbit anti-dog polyvalent reagent) detected autoantibodies against erythrocytes in serum from 53% and 57% of DAT+ dogs, respectively, while other IATs performed less well. Moreover, while the analytic specificity of the IAT methods compared to the DAT ranged from 96-100%, the sensitivity range was only 9-57%. Thus, we still recommend DAT for diagnosis and monitoring of IMHA in dogs but conclude that a positive IAT result may aid diagnostically when serum is available, but fresh red blood cells are not.
RESUMEN
BACKGROUND: A 2019 ACVIM consensus statement on diagnostics for immune-mediated hemolytic anemia (IMHA) in dogs made testing recommendations. As data on the performance of immunohematological tests was lacking, we undertook a comparative analysis. MATERIAL AND METHODS: Anticoagulated blood samples from 126 dogs suspected of having IMHA submitted to a diagnostic veterinary laboratory for a routine direct antiglobulin test (DAT) and from 28 healthy control dogs were evaluated for spherocytosis and autoagglutination before and after three saline washes. Samples were also subjected to different DATs: a gel minitube and an immunochromatographic strip kit used in clinics; neutral gel column cards, microtiter plates (at 4°, 22°, and 37°C), capillary tubes, and flow cytometry used in laboratories. RESULTS: Samples from healthy dogs yielded negative results with all immunodiagnostic tests. Among the 126 samples submitted for DAT 67 were positive by a DAT utilizing microtiter plates with goat anti-dog antiglobulin DAT at 22°C. Notably, DAT results were comparable and consistent across all evaluated methods regardless of antiglobulin and temperature used. DAT+ dogs were more severely anemic and more likely to have erythroid regeneration compared to DAT- dogs. Macroscopic agglutination in tubes or on slides was observed in 48 samples after 1:1 and 1:4 blood to saline dilution, but only persisted in four samples after washing. Among the DAT+ samples, 57% had agglutination, 87% had spherocytosis, and 45% had both. There was good correlation between spherocytosis and DAT results from the six DAT techniques, but the correlation with autoagglutination was only fair. Clinical follow-up was available for 42 dogs. Of the sample from 12 DAT+ dogs collected during treatment, 10 remained DAT+ when tested 1-24 weeks after initial assessment. CONCLUSIONS: Based upon this comparative prospective survey, all in-clinic and laboratory DAT techniques produced similar results when performed by trained personnel and can therefore be recommended for detection of antibody-coated erythrocytes and immunohematological diagnosis. In addition, use of these tests for monitoring response of IMHA dogs to treatment might be valuable.
RESUMEN
Changes in digesta dry matter (DM) and mean digesta particle size (MPS) along the gastrointestinal tract are well known in ruminants, but not in camelids. We collected digesta from the dorsal (d) and ventral (v) first forestomach compartment (C1), the second forestomach compartment (C2), three proximal segments and the subsequent glandular part of the third compartment (C3A-D), the caecum and the faeces twelve llamas (Lama glama). DM analysis indicates the presence of digesta stratification in the C1, the presence of fluid in the C2 to facilitate the sorting function of this compartment, the fluid-absorbing function of the proximal parts of the C3, the secretion of enzymes and digestive acids in the C3D, and the water-resorbing function of the lower intestinal tract. These findings illustrate the functional resemblance between the gastrointestinal tract of camelids and cattle-like ruminants (C1 equivalent to the rumen with stratified contents, C2 to the reticulum, C3A/B/C to the omasum and C3D to the abomasum). MPS analysis revealed a progressive reduction in MPS from the C1 to the distal C3. This gradual transition is different from the clear-cut threshold in ruminants between the reticulum and the omasum and had so far only been described in dromedaries (Camelus dromedarius). These findings indicate that regardless of the convergent property of rumination and resemblance of general mechanisms involved in contents stratification and particle sorting, differences between ruminants and camelids exist that could be interpreted as a more efficient functionality of the ruminant forestomach.