Individual variation in immunosuppressive activity of bovine seminal plasma on concanavalin A-stimulated bovine T lymphocytes in vitro.

Individual dialyzed seminal plasma samples from 21 bulls in routine artificial breeding service were tested for their immunosuppressive activity on in vitro induced blastogenesis of bovine lymphocytes from 3 nonpregnant cows. Concanavalin A was used to induce blastogenesis and thymidine uptake was monitored. Dialyzed seminal plasma from 15 of 21 bulls inhibited deoxyribonucleic acid synthesis response to concanavalin with cow A lymphocytes, and 21 of 21 were immunosuppressive to lymphocytes from cows B and C. Degree of immunosuppression varied according to bull and cow, ranging from 0 to 100%. The lower the percentage response of the lymphocytes to concanavalin stimulation, the less their inhibition by dialyzed seminal plasma. The degree of immunosuppression produced by a bull's dialyzed seminal plasma was not highly related either to amount of protein in the seminal plasma or to the bull's 60 to 90-d nonreturn rate.

Immunosuppressive activity of bovine follicular fluid on bovine T lymphocytes in vitro.

Follicular fluid from bovine follicles at various stages of the estrous cycle and cysts were tested in vitro for immunosuppressive activity. Mitogen-stimulated lymphocyte assays and the mixed lymphocyte reaction were used as test procedures and thymidine uptake was monitored. Follicular fluid and cyst fluid inhibited the deoxyribonucleic acid synthesis response of bovine lymphocytes to mitogenic and antigenic stimulation. The nature of the suppression varied with the mitogen or antigen assay used, with the estrual stage, with size of follicle within the diestrual stage, and with the type of ovarian cyst. Blood sera from estrual or diestrual cows were not immunosuppressive. Heating (56 degrees C for 30 min) follicular fluid from normal follicles and cysts destroyed most of its suppressive activity toward Concanavalin A-stimulated lymphocytes but did not suppress that in luteal cyst fluid. The suppressive activity in follicular fluid toward phytohemagglutinin-stimulated lymphocytes was more stable to heat than that against Concanavalin A-stimulated lymphocytes. Because the immunosuppressive activity in follicular fluid was heat labile, estrogen, progesterone, and other steroids were not the active suppressive molecules. Proposed roles for these immunosuppressive substances are hypothesized in preventing autoantibody formation to the zona pellucida and in the inflammatory reaction leading to ovulation.

Effect of estrual stage on complement activity in bovine follicular fluid.

Ovaries were excised from 112 cows during specific stages of the estrous cycle and from cows with follicular cysts. Follicular fluid was collected from the largest follicle at estrus (d 1), metestrus (d 2 to 4), early diestrus (d 5 to 8), mid diestrus (d 9 to 14), late diestrus (d 15 to 18), proestrus (d 19 to 21), and from follicular cysts. Complement was measured with a standard hemolytic assay. Complement in follicular fluid varied with stage of cycle and was two to five times higher at estrus than at diestrus. Complement in follicular fluid was 5 to 22-fold higher than in blood serum. Blood complement was not influenced by stage of cycle. Cystic follicular fluid had complement amounts similar to that of metestrus follicles. The relationships of complement to ovulation, sperm maturation, fertilization, and sperm removal from the tract are discussed.

Immunosuppressive activity of bovine seminal plasma on bovine lymphocytes in vitro.

Pooled seminal plasma obtained from six bulls was tested for its immunosuppressive activities in vitro on blastogenesis of bovine lymphocytes. Lymphocyte stimulation assay and mixed lymphocyte reaction were used as test procedures. Concanavalin A and phytohemagglutinin were utilized in the lymphocyte stimulation assays as nonspecific stimuli. Tritiated thymidine uptake by the lymphocytes was measured after incubation for 5 d with seminal plasma. Undiluted seminal plasma was cytotoxic to lymphocytes up to and at 1/100 dilution but not at 1/400. Undiluted seminal plasma was immunosuppressive at a 1/400 dilution. Lyophilization of seminal plasma without prior dialysis resulted in loss of immunosuppressive activity. After dialysis and lyophilization, the reconstituted seminal plasma suppressed thymidine uptake by Concanavalin A and phytohemagglutinin-stimulated bovine lymphocytes while being noncytotoxic. This paper documents that bovine seminal plasma is immunosuppressive and also cytotoxic to the bovine cells that generate an immune response.

Identification of an immunosuppressive protein in bovine seminal plasma with activity against bovine lymphocytes.

Seminal plasma was determined to have immunosuppressive activities in vitro on Concanavalin A-induced blastogenesis of bovine lymphocytes. This immunosuppressive activity was isolated and characterized using a pool of seminal plasma from five bulls. The immunosuppressive activity was associated with seminal plasma proteins of less than 50,000 daltons and also with those of approximately 150,000 daltons. The 150,000-dalton proteins were precipitable at 33% saturation with ammonium sulfate and with an antibody to a Sephadex G-200 chromatographic fraction of seminal plasma. The immunosuppressive activity of the less than 50,000 dalton proteins was lost upon chromatography on Sephadex G-25. This suggests that interaction of several components of low molecular weight might be necessary for this inhibitory activity to be expressed.

The influence of some sample handling factors on progesterone and testosterone analysis in goats.

This study validated the use of commercially available radioimmunoassay kits for measuring the circulating progesterone and testosterone levels of goats. Progesterone and testosterone levels were then assayed in plasma which was collected from 23 does and 8 bucks. Collections from each animal were divided into three sodium fluoride-potassium oxalate (F/OX), one heparin, and one EDTA tubes and also into a tube without anticoagulant. Plasma from an F/OX tube was separated immediately from the blood cells by centrifugation. Serum or plasma was also separated after storage for 24 hours with F/OX, heparin or EDTA anticoagulant at 22 degrees C or with F/OX at 5 degrees C. A significant decline in assayable progesterone occurred in samples stored at 22 degrees C with each anticoagulant used and in the serum sample. Samples stored at 5 degrees C for 24 hours with F/OX anticoagulant contained concentrations of progesterone which did not differ significantly from those in samples where plasma was removed immediately. Assayable testosterone did not change with the anticoagulant used or vary with the storage temperature when F/OX tubes were stored at 5 degrees C and 22 degrees C for 24 hours. Results indicate that sample storage does influence levels of measured progesterone but not testosterone in goats. Progesterone assay is best done on plasma which is immediately separated from blood cells or on samples which are stored at 5 degrees C.

Assessment of parameters of physical growth of the newborn in relation to maternal supplementation with iron alone or with some biological additives.

Intrauterine growth charts were used for the assessment of physical growth of newborn infants in relation to maternal supplementation. Iron alone or with other additives (Copper, folic acid, ascorbic acid) were given during different trimesters of pregnancy. The study revealed that iron supplementation with or without other additives has an effective role in improving the haematological pattern in the neonates who also achieved better physical growth particularly with early supplementation with iron and ascorbic acid. Therefore, such supplementation is recommended.

Calcium and phosphorus metabolic changes in children with hepatic bilharziasis.

Investigations were done to study the calcium and phosphorus metabolic changes in 35 children with hepatic bilharziasis, of whom 12 were ascitic. Thirteen normal children served as controls. Despite a normal serum level of both calcium and inorganic phosphate in the Bilharzial group, yet both were increased in urine. The mean values obtained from the ascitic fluid were 70% and 90.2% of their serum levels, respectively. The corrected creatinine clearance was significantly diminished, while PEI was significantly increased, and the % TRP significantly decreased, with still a more marked decrease in the ascitic cases. Following a phosphate lead the serum inorganic phosphate level showed a significantly more rapid lowering in both bilharzial groups, as compared to the control group. On the other hand bilharzial cases responded to the intravenous calcium load by a significant retention of calcium in the serum.