An ELISA was used to detect IgG, IgM, and complement (C3) on the surface of canine erythrocytes. Erythrocytes were placed in wells of a microtitration plate and incubated with affinity purified, alkaline phosphatase-conjugated anti-canine IgG, IgM, or C3. Results of the ELISA were compared with the direct antiglobulin test (DAT) by preparing standard reference curves from canine blood type A erythrocytes that had been incubated with serial dilutions (1:2 to 1:8,192) of canine anti-A serum. The ELISA detected increased erythrocyte-bound immunoglobulin and complement at two- to fourfold dilutions greater than thoe required for positive results with the DAT. The ELISA required small sample and reagent volumes and detected lower concentrations of immune components than did the DAT.

When incubated with solutions of hydrogen peroxide, erythrocytes of stress-susceptible pigs produced more by-products of lipid peroxidation (as measured as thiobarbituric acid-reactive substances [TBARS]) than did erythrocytes from stress-resistant pigs. Using this technique, discrimination between the 2 pig types was absolute at hydrogen peroxide concentrations of 0.9 and 1.5%. This was in contrast to other methods of identifying stress-susceptible pigs, such as osmotically induced erythrocyte lysis and the determination of plasma pyruvate kinase and creatine kinase activities, for which considerable overlap of data was observed between pig types. The increased TBARS production by erythrocytes was further evidence for the existence of an antioxidant abnormality in stress-susceptible pigs. However, because there were no discernible differences in the major blood antioxidant-related values between stress-susceptible and stress-resistant pigs, the nature of the defect remains unclear. The production of TBARS by erythrocytes when incubated with hydrogen peroxide provides an improved method for identifying stress-susceptible pigs.

The ability of Mycoplasma hyopneumoniae to agglutinate RBC was evaluated to develop an in vitro cytadsorption assay. Using swine RBC in a microtitration hemagglutination test, no agglutination or partial agglutination was detected. Comparison of RBC from various other species indicated that improved hemagglutination was obtained with RBC from turkeys. This hemagglutination was detected only when mycoplasma cells used in the assay had been frozen and thawed, heated at 50 C for 30 minutes, or treated with trypsin. Treatment of RBC with trypsin or neuraminidase enhanced hemagglutination. Possible surface lectin activity in M hyopneumoniae was evaluated by use of carbohydrates in a blocking assay; hemagglutination was not inhibited by any of 13 carbohydrates evaluated. Mycoplasma hyopneumoniae convalescent porcine serum and monoclonal antibodies against 2 M hyopneumoniae immunogens of molecular weights of 64,000 and 41,000 inhibited hemagglutination.

The proteins of the bovine erythrocyte membrane were analyzed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow sedimentation rate of bovine erythrocytes were investigated by treating the erythrocytes with trypsin. The erythrocyte sedimentation rates of bovine erythrocytes from Holstein and Korean native cattle were very slow compared with the human one (1/7 as slow as the human one) as reported previously. However, when human and Holstein erythrocytes were treated with trypsin (0.2 and 0.5 mg/ml) for 1 hour at 37deg C, their sedimentation rates were markedly accelerated while the sedimentation rate of Korean native cattle's erythrocytes were not affected. Although the general protein profiles of the bovine erythrocyte membranes were almost similar to that of human, bovine erythrocyte membranes showed one additional protein band called band Q in this study, which migrated electrophoretically to the mid-position between band 2 and band 3 in human erythrocyte membranes. Treatment of Holstein and human erythrocytes with trypsin caused a decrease or disappearance of the band Q from the erythrocyte membrane. Although the band Q in Korean native cattle's erythrocyte membrane was decreased by trypsin treatment of the erythrocytes, the magnitude of the decrement was not so pronounced as in the case of human and Holstein erythrocytes. The glycoprotein profiles of the bovine erythrocyte membranes revealed by periodic acid-Schiff stain showed a marked difference from that of human. The PAS-1 (glycophorin) and PAS-2 (sialoglycoprotein) present in human erythrocyte membrane were almost absent from the bovine erythrocyte membranes. Instead, the bovine erythrocyte membranes showed a strong PAS-positive band near the origin of the electrophorograms, which is named as PAS-B in this study

The frequency of appearance of HI antibody against Japanese encephalitis virus was investigated over the three-year period 1982 through 1984. The sample population consisted of 1306 cattle in Saitama prefecture (located in central Japan), and 536 cattle in Kagoshima prefecture (located in southern Japan). During the study period, there was no significant difference in the average antibody-positive rate between cattle in Kagoshima (68.8 %) and those in Saitama (65.5 %). Variations in the monthly antibody-positive rate and mean monthly antibody titer in the Saitama cattle followed a mountain-like pattern with the peak observed during summer. In the Kagoshima cattle, both parameters also increased in summer and these increases persisted even in winter. The distribution of antibody-positive rate as a function of age ranged from 64.0 % to 82.8 % in Saitama, though no correlation was observed between these two factors. In the Kagoshima cattle, however, the antibody-positive rate increased with age as follows; the values averaged 29.4 % in the one-year-old cattle group, 50.0 % in the two-year-old group, 47.4 % in the three-year-old group and 74.5 % in the four-year-old group. There was no significant correlation between the mean antibody titer and age in either Saitama cattle (15.3-22.5) or Kagoshima cattle (20.0-32.3)

In order to enumerate the T-lymphocytes in bovine peripheral blood lymphocytes (PBL) by E rosette assay, KGRBC were treated with various concentrations of 2aminoethyl-isothiouronium bromide (AET) and dextran (Dex), singly or in combination. To further standardize the assay, optimum concentration of AET-and/or Dex-treatment and incubation time for rosette forming cell (RFC) counts were determined. The levels of B-lymphocytes in the PBL were evaluated by erythrocyte-antibody (EAfc)- and erythrocyte-antibody-complement (EAC)- rosetting techniques. The PBL from 20 clinically normal Korean cattle were formed as low percentage of spontaneous E-rosette (6.7 +- 2.4 %) in control group, whereas in KGRBC treated with 0.1M AET for 20 minutes and 8 % Dex were formed as 37.3 +- 2.7 % and 45.1 +- 2.1 %, respectively. And the synergistic effects were noted no less than 66.5 +- 5.6 % when the KGRBC treated with 0.1M AET and 8 % Dex subsequently and rate of RFR did not change significantly between 3-24 hours incubation time at 4deg C, EA-and EAC-RFR were 23.3 +- 9.1 % and 23.1 +- 7.9 %, respectively. These results suggest that the KGRBC would be a useful agent for the enumeration of T-lymphocytes by E rosette assay and B-lymphocytes by EA-or EAC-rosette assay in cattle-PBL

The study was undertaken to obtain indirect values of volume of packed red cell (VPRC) without centrifugation, using the erythrocyte sedimentation rate (ESR) of diluted blood in dogs. ESRs of diluted blood using the diluent of autologous plasma in which formed numerous RBC-rouleau clumps, autologous serum in which formed a few RBC-rouleau clumps, and 5 % dextrose or 6 % sucrose solutions in which formed numerous RBC-aggregation clumps, were accelerated. But ESR of diluted blood using the 0.9 % saline, D-S, ACD-B, CDP or D-PBS solutions were sluggish, because erythrocytes were dispersed in these diluents. Reliable values of VPRC on the basis of the correlating regressive equation to the ESR could be derived from values of 60deg C-angled-micro-ESR/40 min in the mixture, four parts of 5 % dextrose solution and one part of whole blood. In the ESR values of diluted blood with low ratio, 1 : 1-3 : 1, 60deg C-micro-ESR was higher than 60deg C-Wintrobe-ESR. But, in the diluted blood with high ratio, 4 : 1-5 : 1, there was no different ESR values. For an aid of practical use, authors suggested a list of the 60deg C-micro-ESR/40 min in the diluted blood with equivalent VPRC of whole blood

The proteins of the ruminant erythrocyte membranes were analysed by polyacrylamide gel electrophoresis in sodium dodecyl sulfate, and their relations to the slow erythrocyte sedimentation rate (ESR) of the ruminants were investigated by treating the erythrocytes with proteinases such as trypsin, chymotrypsin and pronase, and glycosidases such as neuraminidase and galactosidase. Protein content in the erythrocyte membrane was 2.85 +- 0.28 in human, 3.60 +-0.41 in Korean cattle, 3.71 +- 0.36 in Holstein, 4.13 +-0.83 in Korean native goat and 3.94 +- 0.56 mg/ml in sheep, showing higher in ruminant animals than in human (p0.01). Although the general protein profiles of the ruminant erythrocyte membranes were almost similar to that of human, all the ruminant erythrocyte membranes showed one additional protein band, called band- Q in the previous report on proteins of bovine erythrocyte membrane, which migrated electrophoretically to the mid position between band-2 and band-3 in human erythrocyte membranes. The glycoprotein profiles of ruminant erythrocyte membranes revealed by periodic acid Schiff (PAS) stain showed a marked difference from that of human. The PAS-l (glycophorin) and PAS-2 (sialoglycoprotein) present in human erythrocyte membranes were almost absent from the ruminant animals. Instead, a strong PAS-positive band near the origin of the electrophorograms, which was named as PAS-B in the previous report on proteins of bovine erythrocyte membranes, was shown in the ruminant animals except sheep. In addition, the erythrocyte membranes of Korean native goat and sheep showed a moderate PAS-negative band near the tracking dye of the electrophorograms, which was named as PAS-G in this study. In the erythrocyte treated with the enzymes, the migration of each protein fracture of erythrocyte membranes in response to each enzyme was diverse according to different species or breed of ruminant animals. Among others, band-Q present in ruminants was slightly or moderately decreased by trypsin-, chymotrypsin-, and pronase- treatments of the erythrocytes, but not only in sheep