Blood oxygen transport and hemoglobin function in three tropical fish species from northern Australian freshwater billabongs

The oxygen binding properties of whole blood and hemoglobin were studied in three phylogenetically distant tropical teleosts inhabiting freshwater billabongs that exhibit marked seasonal oxygen stratification. The water breathing saratoga (Scleropages jardinii, Family Osteoglossidae), derived from an ancient divergence of the stem line leading to all other extant teleosts, has a low blood oxygen carrying capacity, Bohr and Root effect, and a high blood oxygen affinity. Erythrocytes contain a single major hemoglobin component, and equimolar concentrations of ATP and GTP. The tarpon (Megalops cyprinoides, Family Megalopidae), a facultative air-breather, has the highest blood oxygen carrying capacity and Bohr effect, and a low blood oxygen affinity and Root effect. Erythrocytes contain a single major hemoglobin and ATP as a putative regulator of hemoglobin oxygen affinity. Barramundi (Lates calcarifer, Family Centropomidae), is an obligate water-breather with intermediate blood oxygen binding properties, and the smallest Root effect. Erythrocytes contain at least 7 hemoglobins, and equimolar concentrations of ATP and GTP. Functional properties of these three blood oxygen transport systems are considered in terms of the respiratory environment and demand for oxygen. Our interpretation supports the hypothesis that the process of speciation can lead to divergence in physiological mechanisms, irrespective of past or present selection pressures.     

Cell titration assay for measuring blastogenesis of bovine lymphocytes

The blastogenic response of bovine peripheral blood lymphocytes to phytohemagglutinin (PHA) and to microbial antigens was measured using a lymphocyte titration assay. Culture conditions, including lymphocyte concentrations, incubation periods and medium formulation, were established which produced linear or nearly linear responses over a range of cell concentrations. These conditions were established by testing lymphocytes from unimmunized cattle and from heifers infected with Brucella abortus with PHA and a B. abortus extract. Four cell concentrations in 2-fold increments were selected for measuring responses to PHA (3.125 X 10(3) to 2.5 X 10(4) cells/well) and to antigens (5.0 X 10(4) to 4.0 X 10(5) cells/well). The strength of response varied among animals and also over time for individual animals, but the titration assay allowed exponential proliferation to be distinguished from decline, which may have been due to overcrowding of microtiter wells, exhaustion of nutrients or induction of regulatory events. This assay provided a more reliable and discriminating method of evaluating lymphocyte proliferation responses than that achieved by single point assays. The displacement of the titration curves could be used to estimate the relative frequency of lymphocytes responding to antigens or mitogens.     

Technical note: isolation and characterization of sheep ruminal epithelial cells

A system for the isolation and characterization of sheep ruminal epithelial cells has been developed. Ruminal papillae from the ventral cranial sac of 12- to 24-wk-old Dorset ram lambs were subjected to serial tryptic digestions. Initial digestions (two 15-min periods) in the series were used to remove keratinized cells of the stratum corneum, which were able to produce only small amounts of beta-hydroxybutyrate from butyrate (5.37 +/- .72 nmol/120 min/per milligram of dry cell weight). Later digest fractions, containing primarily cells from the stratum basale, exhibited high viabilities (75 to 95%) and proved capable of converting butyrate to beta-hydroxybutyrate at relatively high rates (33.6 +/- 6.7 nmol/120 min per milligram of dry cell weight). Neither acetate nor propionate underwent significant conversion to beta-hydroxybutyrate. However, addition of acetate inhibited (77.4% of control) and addition of propionate stimulated (200% of control) beta-hydroxybutyrate production. Acetate addition reduced the propionate-induced stimulation of beta-hydroxybutyrate production from butyrate (136% of control). These results are similar to those obtained from in vitro incubations of ruminal papillae and suggest that an isolated cell system may prove useful in the further study of ruminal epithelial metabolism.     

Seroprevalence and comparison of isolates of endometriotropic bovine herpesvirus-4.

Sixty-eight cases of suppurative, ulcerative endometritis associated with Bovine Herpesvirus-4 (BHV-4) in postparturient dairy cows (62 Holsteins and 6 Jerseys, mean age 4.2 years) were confirmed by a combination of histopathology, fluorescent antibody assays, electron microscopic evaluation of uterus, and polymerase chain reaction (PCR). All cases occurred in the 3- to 28-day postpartum period, and histologic lesions among various cows were consistent when compared with postpartum interval. The endometrial lining epithelium was necrotic and ulcerated from 3 to 7 days postpartum, with only mild inflammation in the lamina propria and submucosa. From 1 to 4 weeks postpartum, the ulcers were confluent to diffuse. Epithelium was replaced by fibrinonecrotic, suppurative mats, resulting in severe bacterial pyometra by day 24. Seroprevalence to BHV-4 in one dairy with a history of 18 mortality cases was 36% (107 of 296). In a random sample of 8 cows from this herd, none had serologic titers in blood sampled 2 weeks prepartum, but 3 of 8 seroconverted with significant titers of 1:8 to 1:16 at 2 weeks postpartum. By 10 weeks postpartum, all 8 cows returned to negative serologic status. Two of 6 cats from the premises also had positive titers. Random serum samples taken from 480 dairy cattle at sale barns indicated 76 (16%) were positive by serum neutralization. Clinical signs, postparturient timing, and histologic lesions were very similar to those previously reported in Belgium with BHV-4. But sequence analysis of PCR products of the glycoprotein B region of 4 separate field isolates of endometriotropic BHV-4 suggests these field isolates were more closely related to the North American nonvirulent strain DN-599 than to the endometriotropic European strain V.     

Animal performance, plasma hormones and metabolites in Holstein and Belgian Blue growing-fattening bulls

Six Holstein (light-muscled type) and six Belgian Blue bulls (double-muscled type) were fed a finishing diet. Average daily gain was 1.36 kg for the Holstein bulls vs 1.24 kg for the Belgian Blue bulls (P less than .05). Holstein bulls consumed more feed (2.3 vs 1.8 kg/100 kg body weight, P less than .001) than the Belgian Blue bulls. The dressing percentage (55.4 vs 65.8%, P less than .001) and the proportion of muscle (56.1 vs 71.3%, P less than .001) in the carcass were less, whereas the proportions of adipose tissue (28.3 vs 15.4%, P less than .001) and bone (15.7 vs 13.4%, P less than .05) were higher in the Holstein bulls. Plasma creatinine determined in samples obtained once a week was lower (11.0 vs 20.3 mg/liter, P less than .001) in the Holstein bulls. In contrast, Holstein bulls tended to produce more triiodothyronine (2.3 vs 1.8 nM, P less than .10), tetraiodothyronine (71.9 vs 54.7 nM, P less than .10) and insulin-like growth factor I (IGF-I; 340 vs 205 ng/ml, P less than .20) than the Belgian Blue bulls. Growth hormone, insulin, IGF-I and testosterone were measured at 20-min intervals during two 24-h periods. In wk 6, Holstein bulls tended to produce more growth hormone than the Belgian Blues, as indicated by higher total peak area (3,185 vs 2,431 ng), peak amplitude (34.1 vs 22.6 ng/ml, P less than .10) and baseline (4.6 vs 3.3 ng/ml, P less than .20). In wk 27, the trends were opposite.(ABSTRACT TRUNCATED AT 250 WORDS)