A Potential Screening System for Identifying Sorghum Ecotypes with Increased Lysine in the Seeds

Classical “high-lysine” sorghum lines are characterized by smaller seeds than average, due to a decrease in prolamin synthesis and a subsequent decrease in yield. To exploit the natural variation in lysine content and to identify ecotypes with a seed lysine content higher than average, characterized by plump seeds, a method was developed based on root-growth inhibition of seeds growing on a medium containing aminoethylcysteine (AEC), a lysine analogue. By using a collection of sorghum mutants and ecotypes a correlation coefficient of 0.926 between root length and lysine content was established. This method, which uses the root length of plants growing on aec to indicate which lines have a potential elevated lysine content, can be applied for the screening of sorghum germplasm. Since this is a non-destructive method it can also be used at the individual seed level, for example for screening progenies of regenerated plants from in vitro culture to exploit the somaclonal variation.     

The diagnosis of nephrogenic diabetes insipidus in the dog]

Nephrogenic diabetes insipidus is characterized by polyuria, hyposthenuria and compensatory polydipsia. With conventional clinical and laboratory examinations the diagnosis can be established and the differential diagnostic diseases excluded. In order to verify the diagnosis, differentiate the condition from central and nephrogenic diabetes insipidus and rule out psychogenic polydipsia, additional laboratory studies are required: of these, the concentration test, modified Carter-Robbins test and AVP stimulation test are described.     

Intradermal testing of horses with chronic obstructive pulmonary disease and recurrent urticaria.

Six horses with chronic obstructive pulmonary disease (COPD) and 8 horses with recurrent urticaria were skin tested with 67 extracts from 58 allergens, including pollens, epidermals, cultivated farm plants, dusts, molds, and insects. Reactions were evaluated 3 times over a 24-hour period immediately after the injections. Results were compared with those obtained from 11 clinically normal horses. All horses had positive skin test reactions. Significant differences was evident between horses with COPD and clinically normal horses for only 3.0% of the possible extract reactions, and between horses with urticaria and clinically normal horses for only 4.5% of the possible extract reactions. Horses with COPD or urticaria had greater total percentage of allergen extract reactions than did clinically normal horses. Positive reactions were observed at all 3 evaluation periods, and late-onset reactions were not always preceded by positive reaction at earlier periods. All horses with COPD or urticaria had at least 1 skin test reaction that exceeded the mean +/- 2 SD, as calculated for each of the 67 extracts for the group of clinically normal horses.     

Analysis and pharmacokinetics of cimaterol in growing Holstein steers.

Pharmacokinetic parameters for the beta 2-adrenergic agonist, cimaterol (CIM), were determined in growing Holstein steers. Compartmental analysis was used after measurement of CIM in body fluids by affinity chromatography and HPLC using UV detection. Recoveries from spiked plasma and urine standards were 70 +/- 1.2% and 68 +/- 1.1%, respectively. The minimum detection level in plasma was 1 ng/mL and the average CV was 5.1% for concentrations that ranged from 1 to 30 ng/mL. Four steers (276 +/- 24 kg) received 15 mg of CIM by bolus intravenous injection. Plasma CIM levels declined in a biphasic manner with half-lives of 2.5 min for the distribution phase and 54 min for the elimination phase. A two-compartment open model was used to describe the disappearance of CIM and the following pharmacokinetic parameters were obtained: central compartment volume (Vc) = .76 L/kg, apparent volume of distribution (Vd) = 4.1 L/kg, and transfer rate constants from the central to peripheral compartment (k12) = .177/min, from the peripheral to central compartment (k21) = .054/min and elimination from the central compartment (kel) = .074/min. After 8 h, total urinary CIM accounted for only 18.3% of the administered dose. Results suggest that circulating concentrations of CIM in growing steers are influenced by its accumulation in an unidentified peripheral pool and its conversion into unknown metabolite(s) before elimination.     

Release of tumor necrosis factor-alpha from bovine alveolar macrophages stimulated with bovine respiratory viruses and bacterial endotoxins

The release of tumor necrosis factor-alpha (TNF-) from cultured bovine alveolar macrophages (BAM) was evaluated following stimulation of BAM with bovine herpesvirus-1 (BHV-1), parainfluenza-3 (PI-3) virus, bovine respiratory syncytial virus (BRSV), Escherichia coli 0111:B4 endotoxin, Pasteurella haemolytica type 1 endotoxin, Pasteurella multocida endotoxin, and virus/endotoxin combinations. A cytotoxic assay system using Georgia bovine kidney cells as targets was used to measure TNF- activity. The cytotoxic activity was neutralized by an anti-human TNF- monoclonal antibody.

Stimulation of BAM with 1 median tissue culture infectious dose (TCID₅₀) of live or ultraviolet (UV)-inactivated PI-3 virus/cell resulted in release of TNF- in significantly (P<0.05) higher amounts than sham-induced BAM. The quantities of TNF- released after live or UV-inactivated BHV-1 or BRSV induction were not significantly higher than sham-induced BAM. E. coli 0111:B4, P. haemolytica type 1 and P. multocida endotoxins stimulated TNF- release in a dose-dependent manner. Sequential exposure of BAM to 1 TCID₅₀ per cell of either live BHV-1, PI-3 virus or BRSV and then 5 μg ml⁻¹ of either E. coli 0111:B4, P. haemolytica type 1 or P. multocida endotoxin caused a significant (P<0.05) reduction in detectable TNF- in seven of nine virus/endotoxin combinations tested, when compared with 5 μg ml⁻¹ of endotoxin alone. Parainfluenza-3 virus/endotoxin combinations stimulated higher TNF- release when compared with other virus/endotoxin combinations. Five out of six test animals had serum-neutralizing antibodies to PI-3 virus, one out of six had serum-neutralizing antibodies to BHV-1, and two out of six had serum-neutralizing antibodies to BRSV, suggesting a possible relationship between serum neutralizing antibodies and TNF- release from in vitro cultivated BAM.     

Potential of a recombinant antigen as a prophylactic vaccine for day-old broiler chickens against Eimeria acervulina and Eimeria tenella infections.

A genetically engineered Eimeria tenella antigen (GX3262), produced as a fusion protein with beta-galactosidase and identified with a monoclonal antibody, induced partial but significant protection in young broiler chickens against experimental E. tenella and Eimeria acervulina infections. The antigen appears to share a T-helper cell epitope with the parasite as evidenced by (a) booster inoculation with either the recombinant antigen or with a small number of live oocysts enhanced the protective immunity in GX3262 primed chickens, and (b) ability of the antigen to induce in vitro stimulation of T-cells from chickens immunized with antigen or parasite. These observations suggest the feasibility of a single vaccination of 1 or 2-day-old broilers with GX3262 to induce an acceptable degree of protective immunity. The implications of the observations reported here are far reaching in terms of a practical coccidiosis vaccine for poultry, and show for the first time that 1-day-old broiler chickens can be efficiently vaccinated with a recombinant antigen against one or more species of Eimeria.     

Plasma triglyceride concentration during intravenous infusions of propofol and Intralipid in sheep

Plasma triglyceride concentrations were measured in sheep given Intralipid or propofol, which is carried in a vehicle very similar to 10% Intralipid. A bolus dose was administered followed immediately by an infusion of the same agent for 2 h. In the animals that received propofol, the measured concentration increased by a mean amount of 3.39 mmol/l when the infusion rate was l ml/min (Group Pl) and by 7.13 mmol/l when it was 2 ml/min (Group P2). When 10% Intralipid was administered and infused at 1 ml/min (Group I10), the measured concentration increased only by 0.95 mmol/l. One hour after stopping the infusion, the excess of measured concentration over baseline had decreased in the Pl and I 10 groups to 0.52 and 0.13, respectively, of the corresponding maximum excess. The method adopted for measuring plasma triglycerides is widely used in hospitals; however, an incidental observation revealed that it is inappropriate in the presence of injections of propofol or Intralipid. Despite this, evidence and argument are presented to support the conclusion that, with propofol, plasma triglyceride concentrations increased more rapidly during the infusions and returned to baseline more slowly than with a corresponding amount of Intralipid.     

Studies on efficacy and stability of a vaccine bait containing ERA strain of rabies virus propagated in a BHK-21 cell line.

In a dose response study in foxes, the median protective dose of ERA BHK21 vaccine in a blister pack bait was 10(6.0) tissue culture infective doses (TCID)/mL, while artificially aged baits with titers of 10(6.3) TCID/mL induced seroconversion in 78% of foxes. There was no significant difference in the development of antibodies in foxes receiving 1, 2 or 3 mL volumes of vaccine in the bait. When baits were exposed to the elements and fed to foxes over a 21 day period, 85% of the animals seroconverted. Age, sex and the way in which the vaccine container was contacted did not appear to be factors in the responses of these animals. Juvenile foxes, approximately six months of age, were marked more readily with the tetracycline bait marker than older animals. Approximately 25% of foxes did not appear to respond well to vaccination and the titer of the vaccine was a critical factor in producing seroconversion in these animals.