Use of monoclonal antibodies against human HLA II antigens for the detection of bovine B lymphocytes and macrophages]

The crossreactivity of mouse monoclonal antibodies (MoAbs) (Tab. I) prepared against human HLA-DR and HLA-DP antigens was studied in various bovine cells: lymphocytes from lymph nodes and peripheral blood, adherent (B) and nonadherent (T) lymphocytes, monocytes, granulocytes and platelets. In the immunofluorescence test, MoAbs Bra13, Bra14, Bra20, Bra22, Bra30, Bra70, HL-38 reacted with bovine B lymphocytes and monocytes, but not with other tested cells (Tab. III, IV). These antibodies, except Bra22, were positive with B lymphocytes in the complement dependent cytotoxic test (Tab. II). The similarity of the bovine antigens and HLA-DR antigens determined by used MoAbs was also proved by immunoblotting. Monoclonal antibodies Bra38 and BraFB6 did not react with the bovine cells and separated antigens. The epitope (HLA-DR) recognized by the antibody Bra38 is probably absent in cattle. The presence of HLA-DP analogue determined by the antibody BraFB6 has not been confirmed. The crossreactive MoAbs could be used for the detection of B lymphocytes and macrophages in veterinary immunology.     

Bovine respiratory syncytial virus-specific immune responses in cattle following immunization with modified-live and inactivated vaccines. Analysis of the specificity and activity of serum antibodies.

Cattle were immunized with vaccines containing modified-live or inactivated bovine respiratory syncytial virus (BRSV) and serum antibody responses were analyzed. Compared with preinculation values, at Day 14 after two biweekly immunizations with modified-live or inactivated vaccines there were significant increases in BRSV-specific titers in the sera of cattle that received both types of vaccines, as determined by a whole cell ELISA. Using a blocking ELISA and radioimmune precipitation it was determined that there was recognition of the fusion (F) protein by antibodies from cattle that received both types of BRSV antigens: however, virus neutralization assays revealed that only cattle that received modified live virus, either in monovalent or polyvalent vaccines, developed neutralizing antibodies to BRSV after two immunizations. These results indicate that inactivation of BRSV can lead to a dissociation between serological recognition of the F protein and virus neutralization in vaccinated cattle.     

Evaluation of a monoclonal blocking ELISA and IHA for antibodies to Mycoplasma hyopneumoniae in SPF-pig herds.

A monoclonal blocking enzyme-linked immunosorbent assay (ELISA) and an indirect haemagglutination assay (IHA) were applied to serum samples from 124 specific pathogen-free (SPF) breeding and multiplying herds, which participate in the routine serological surveillance of the Danish SPF programme. Clinical and pathological observations of the herds and microbiological culturing of Mycoplasma hyopneumoniae were used to calculate herd sensitivity, herd specificity and herd predictive values for the two serological assays. The ELISA was superior to the IHA in herd sensitivity and herd specificity, with values of 93 per cent and 96 per cent, respectively, for the ELISA, and 61 per cent and 92 per cent for the IHA. During the six month period of evaluation 2.5 per cent of the herds were infected with M hyopneumoniae each month. At this level the IHA was found to have a positive herd predictive value of 16 per cent, compared with 39 per cent for the ELISA. The negative herd-predictive value on the same level was 99.8 per cent for the ELISA and 98.9 per cent for the IHA. If the assays were applied to a group of herds with a herd prevalence of M hyopneumoniae infection of 30 per cent (as is the case with the production herds in the Danish SPF programme) the predictive value of a positive herd diagnosis would be 91 per cent for the ELISA and 76 per cent for the IHA, and the predictive value of a negative herd diagnosis would be 97 per cent with the ELISA and 85 per cent with the IHA.(ABSTRACT TRUNCATED AT 250 WORDS)     

Use of progesterone in microspheres for maintenance of pregnancy in mares.

Administration of progesterone in poly(d-,l-lactide) microspheres was used to maintain pregnancy in mares after luteolysis was induced by treatment with prostaglandin F2 alpha at day 14 of pregnancy. Mares were given vehicle only (control, n = 6) or 0.75 g (n = 7), 1.5 g (n = 8), or 2.25 g (n = 5) of microencapsulated progesterone at days 12 and 22 of pregnancy. Serum progesterone concentrations were determined daily, and pregnancy was evaluated by transrectal ultrasonography on alternate days. Significantly (P less than 0.05) more mares given 1.5 or 2.25 g of progesterone (6 of 8 and 4 of 5 mares, respectively), but not those given 0.75 g (3 of 7 mares), maintained pregnancy through day 32, compared with control mares (0 of 6). Progesterone concentrations decreased significantly (P less than 0.025) in all groups after administration of prostaglandin F2 alpha at day 14, and significant (P less than 0.05) effects of time and treatment on progesterone concentrations were found between days 12 and 22, and 22 and 32. Although treatment with 1.5-g and 2.25-g doses of microencapsulated progesterone improved maintenance of pregnancy, compared with that of vehicle-treated controls, administration of 2.25 g of microencapsulated progesterone appeared to be most efficacious in maintenance of pregnancy during the study interval.     

Ovarian follicular characteristics, embryo recovery, and embryo viability in heifers fed high-fat diets and treated with follicle-stimulating hormone.

Postpubertal beef heifers (n = 55) were used to examine the effects of high-fat diets, independently of energy intake, on nonesterified fatty acid and lipoprotein metabolic patterns, ovarian follicular dynamics, and embryo recovery/viability after FSH superstimulation. High-lipid (HL) diets (5.4% added fat) increased (P < .01) serum concentrations of cholesterol, but not of nonesterified fatty acids, during the 35-d period before FSH treatment. Development of medium-sized (5 to 9.9 mm) follicles was enhanced (P < .05) during this period in heifers fed the HL diet. The HL diet increased total cholesterol (P < .05) and progesterone (P = .14) concentrations in follicular fluid obtained at ovariectomy (n = 10) 60 h after the onset of FSH treatment, but neither estradiol-17 beta nor androstenedione was affected. Granulosa cells recovered from FSH-induced, estrogen-active follicles in heifers fed the HL diet produced greater quantities of progesterone (P = .06) and less estradiol-17 beta (P < .05) in vitro than did granulosa cells from heifers fed the normal lipid diet. Dietary treatment did not influence FSH-stimulated recruitment of medium and large follicles, number of ovulations, embryo recovery, or embryo viability. Data suggest that increments in dietary fat intake can alter specific aspects of ovarian steroidogenic potential and can increase the population of medium-sized follicles theoretically available for maturation and harvest during the estrous cycle. However, conditions that limited the latter process in the current experiment are not understood and require further investigation.     

Kinetics of hydration and effect of liquid uptake on specific gravity of small hay and silage particles.

Kinetics of hydration of ground hay and silage particles (2-mm screen), determined by a pycnometric technique, was best described by a two- and one-pool exponential model, respectively. Fractional rates of hydration of the large pool, detected in hay particles only, and of the small pool present in both hay and silage particles averaged .135 and .021 min-1, respectively. When hydration was complete, liquid associated with particles averaged 1.16, 1.90, and .83 g/g of insoluble DM for bromegrass hay, alfalfa hay, and alfalfa silage, respectively. Functional specific gravity, which accounts for the effect of associated gas volume, averaged 1.54, 1.46, and 1.54, but unit specific gravity, calculated to include the effect of gases and liquid of hydration, averaged 1.22, 1.14, and 1.26 for bromegrass hay, alfalfa hay, and alfalfa silage, respectively. Preservation of forage as silage not only lowered gas volume, but also reduced water-holding capacity, both of which contribute to greater unit specific gravity and faster rate of escape from the rumen. In addition, estimates of unit specific gravity of approximately 1.2 indicate that even in the absence of associated gas, hydrated forage particles would tend to escape the rumen at a slower rate than that achieved by more dense particles.     

Function of neutrophils and chemoattractant properties of fetal placental tissue during the last month of pregnancy in cows.

Neutrophils were isolated from the blood of pregnant cows on days 255, 265, and 275 of pregnancy, and on the day of parturition (n = 5/group), and in addition, simultaneously from 4 ovariectomized healthy cows (control animals). Neutrophils were subjected to neutrophil function assays (chemotaxis against zymosan-activated serum, random migration, ingestion of 125I-iododeoxyuridine [IdUR]-labeled Staphylococcus aureus, iodination of proteins, cytochrome C reduction, antibody-independent and antibody-dependent cell-mediated cytotoxicity). Results were expressed as percentage of control animals. Fetal placental tissue (cotyledon), uterine wall tissue, and skeletal muscle were obtained from the principal animals on the aforementioned days via laparotomy, and tissue suspensions were prepared. Chemotaxis of neutrophils was tested against tissue supernatants. Compared with day 255, there was an increase in ingestion of 125I-IdUR-S aureus at parturition, whereas iodination of proteins and cytochrome C reduction were reduced on the day of calving. The other neutrophil functions tested did not change over time of gestation. Fetal placental and uterine wall tissue attracted neutrophils with uterine wall tissue having a tendency to be more potent than cotyledonary tissue. Skeletal muscle tissue did not attract neutrophils. There was no change in chemotaxis response of neutrophils evoked by intrauterine and uterine tissues over time of gestation. It was concluded that at parturition, neutrophil function is impaired with respect to their bactericidal effects, which may render the animal more susceptible to bacterial infections, and that the chemoattractant properties of fetal placental and uterine wall tissues are tissue-specific, at least when compared with skeletal muscle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Surgical treatment of dorsal cortical fractures of the third metacarpal bone in thoroughbred racehorses: 53 cases (1985-1989).

Between January 1985 and May 1989, 53 Thoroughbred horses (mean age 3.2 years) were surgically treated for dorsal cortical fractures of the third metacarpal bone (MC III). All horses were treated with cortical drilling through the fracture line (osteostixis). Diagnosis of the fractures was confirmed by xeroradiography. Lifetime racing records were obtained for all horses. Forty-seven horses returned to racing after surgery (89%). The mean time between surgery and the first race was 6.8 months. Horses had a mean of 10.9 starts before surgery and 16.1 starts after surgery. The mean earnings per start before surgery was $6,459 and after surgery was $5,685. Of the 47 horses that raced after surgery, 70% raced at the same class or improved. Complications related to surgery were seen in 10 horses. Two horses had a second fracture of MC III at the same site, and were again treated by osteostixis, after which both horses returned to competition. Fractured drill bits were left in the MC III of 4 horses. One of these horses had catastrophic failure of MC III. Two horses developed subcutaneous infections and 2 horses had catastrophic failure of MC III in the surgically treated limb. Osteostixis appears to be an effective treatment for returning horses affected with dorsal cortical fractures to racing.     

Analysis of glycoprotein I (gI) negative and aberrant pseudorabies viral diagnostic isolates.

Glycoprotein I (gI) phenotypes and genotypes of 4 pseudorabies viral diagnostic isolates were evaluated by use of in vitro DNA amplification, monoclonal antibody binding, gI-specific serodiagnostic responses, and in vivo virulence approaches. Three viruses were avirulent and did not elicit gI-specific serologic responses, react with gI-specific monoclonal antibodies, or contain gI epitope-encoding DNA sequences. The fourth virus was virulent and did elicit a gI-specific serodiagnostic response. Compared with reference virulent pseudorabies viruses, however, the fourth isolate had reduced reactivity with a group of gI monoclonal antibodies and had a single nucleotide sequence substitution with a corresponding putative amino acid change in the epitopically dominant portion of the gI molecule. Presumably, the first 3 isolates represented diagnostic recoveries of viruses derived from gI-deleted modified-live pseudorabies viral vaccines, whereas the fourth isolate was a virulent but gI-aberrant wild-type virus. Thoroughly assessing the gI status of pseudorabies viral diagnostic isolates was considered to be essential in evaluating the epidemiologic importance of these viruses and in monitoring the validity of gI-based vaccine companion tests now used worldwide in pseudorabies control and eradication programs.     

Pulmonary artery and aortic pressure changes during high intensity treadmill exercise in the horse: effect of frusemide and phentolamine.

Intravenous frusemide (1.0 mg/kg bwt) or phentolamine (0.33 mg/kg bwt) was given to 7 horses 1 h before exercise and their effects on pulmonary artery and aortic pressure changes during strenuous exercise were examined. Short-term near-maximal treadmill exercise (10 m/sec, 3 degrees incline) produced increases in heart rate, mean pulmonary artery pressure (PAP), mean aortic pressure (AP), and packed cell volume (PCV). Frusemide did not affect heart rate, PAP or PCV during exercise. Frusemide significantly decreased mean AP by 10 to 15 mmHg during exercise. Phentolamine produced an increase in heart rate relative to control only early in exercise but not during later, more strenuous, exercise. Phentolamine had no statistically significant effect on AP, PAP, or PCV, but a significant reduction was observed between 180 and 230 sec of exercise when PAP and AP were standardised against heart rate. Frusemide did not prevent horses from haemorrhaging during exercise in this study. Treatment with phentolamine did not sufficiently reduce the PAP and AP to test our hypothesis that a reduction in PAP and AP would eliminate EIPH.