A study of the interaction between bromocresol green dye and bovine,ovine and equine serum globulins

In human medicine it has been shown that the bromocresol green (BCG) dye-binding method for the determination of serum albumin is not entirely specific, the dye reacting also with certain human serum globulins. This causes over-estimation of albumin when reaction times are prolonged beyond 30 seconds.In the present study, serum albumin values obtained from three animal species by the immediate, i.e. less than 30 seconds, BCG reaction were compared with those by the 10-minute BCG reaction. Albumin-depleted sera were prepared using an affinity chromatography technique and their reactions and those of purified gamma globulin preparations with the dye were studied.In cattle, sheep and horses, serum albumin values obtained by the 10-minute reaction were higher than those obtained by the immediate BCG reaction, the differences being statistically significant. Purified gamma globulin did not react with the BCG dye after 10 minutes, but other globulins did. There were differences between the species in the magnitude of the reaction of their globulins with BCG dye.     

Levels of selenium in blood and tissues associated with some selenium deficiency diseases in New Zealand sheep

Whole bloods from weaned lambs with severe selenium responsive unthriftiness usually contain <5 ng Se/ml. Mildly or moderately affected lambs have blood levels of 5-10 ng/ml. Selenium responsive infertility in ewes appears to be associated with whole blood selenium levels below 10 ng/ml.     

Enzyme-linked immunosorbent assay for detection of equine infectious anemia antibody to purified P26 viral protein

An indirect enzyme-linked immunosorbent assay (ELISA) was developed for the detection of equine infectious anemia (EIA) antibody in horse sera. Purified P26 viral protein was the antigen; alkaline phosphatase linked to rabbit anti-horse immunoglobulin G was the conjugate. The ELISA detected EIA antibodies in horse sera as early as 11 to 14 days after experimental inoculations. There was full agreement between the results of ELISA and the agar-gel immunodiffusion tests on EIA proficiency test sera. The ELISA readily detected EIA antibody in horse sera that had weak positive reactions on agar-gel immunodiffusion.     

Receptor-dependent and independent degradation of insulin by isolated swine adipocytes at 37 C

Insulin binding and degradation were measured at 37 C in isolated swine adipocytes. In preliminary experiments, binding decreased rapidly with increasing incubation time. This was associated with a marked increase in insulin degradation. Insulin binding was suppressed by some lots of bovine serum albumin (BSA), which suggests that some commercial preparations of BSA are contaminated with insulin-like molecules. In adipocyte suspensions, greater than 90% of the insulin degraded was due to a nonreceptor mediated process (i.e., insulin degrading activity present in the media). Despite the presence of insulin degrading activity in the media the cells were metabolically (as judged by lipogenic capacity and lactic dehydrogenase activity) and morphologically (greater than 98% excluded trypan blue) intact indicating that the cells were not leaking during the incubation. In subsequent experiments it was found that the specific step associated with transfer of cells during adipocyte isolation resulted in the release of insulin degrading activity. Implementation of a 30-min preincubation and washing sequence after adipocyte isolation removed the media insulin degrading activity, resulting in a marked reduction (approximately 70%) of insulin degradation by adipocyte suspensions. As a result of this modification, binding of tracer quantities of insulin attained steady-state binding conditions and maintained this for 2 h. These results demonstrate that techniques can be used to minimize nonreceptor mediated insulin degradation in adipocyte suspensions. As a result in vitro studies can be conducted that measure insulin binding and biological action in swine adipocytes at physiological temperatures.     

Seasonal transmission of Fasciola hepatica to cattle in northwestern United States

Sentinel steers were placed with 3 beef herds on irrigated pastures in southern Idaho for 1-month periods from May until November 1982 to determine the transmission pattern of Fasciola hepatica. Transmission was found to increase through the pasture season, reaching a peak during November. Overwintering of metacercariae or snail-borne stages was not found to contribute to infections in the year under study. A variety of species of Lymnaea were found to be available in southern Idaho as potential intermediate hosts. The enzyme-linked immunosorbent assay was found to be a good serologic indicator of light infections with F hepatica. The serum gamma-glutamyltransferase activity was not diagnostically significant when the degree of fluke infection was low.     

Visceral leishmaniasis in the German shepherd dog. II. Pathology

Three German shepherd dogs were inoculated with Leishmania chagasi and three with Leishmania donovani and the infection was followed for 82 days. All infected dogs developed splenomegaly and lymphadenomegaly. In lymph nodes there was a reduction in lymphocyte population in paracortical areas, extensive proliferation of macrophages in paracortical areas and medullary cords, follicular hyperplasia, and increased numbers of plasma cells. The spleen had decreased numbers of lymphocytes in periarteriolar lymphoid sheaths, proliferation of macrophages in these regions, follicular hyperplasia, and enlargement of the red pulp with clusters of macrophages and plasma cells. The morphology of the tonsil was similar to the lymph nodes. Clusters of macrophages, often containing Leishmania spp, were present in liver, bone marrow, lung, and the intestines. The morphologic changes in lymph nodes and spleen were suggestive of a suppressed cell-mediated immunity and an active humoral immunity. The German shepherd dog may be a useful laboratory model for the study of immunopathologic changes in visceral leishmaniasis.