In vitro expression and inhibition of procoagulant activity produced by bovine alveolar macrophages and peripheral blood cells

Local and systemic activation of coagulation is frequently associated with bacterial sepsis. The coagulopathy is due, at least in part, to expression of tissue factor (TF) by monocytes and macrophages. The purpose of this study was to evaluate the expression of procoagulant activity by bovine alveolar macrophages, leukocytes and platelets, and to determine the relative potency of three chemical inhibitors of TF expression (pentoxifylline, retinoic acid, and cyclosporin A). Bovine alveolar macrophages were stimulated with lipopolysaccharide (LPS) derived from Pasteurella haemolytica or recombinant bovine tumour nervous factor (TNF) and dose- and time-dependent effects on TF expression were studied. LPS and TNF induced TF expression in alveolar macrophages and LPS treatment of whole blood induced TF expression in mononuclear cells. Neutrophils and platelets also expressed procoagulant activity, but this activity was not inhibited by anti-bovine TF monoclonal antibody. Pentoxifylline (40 mol/L), retinoic acid (0.01 mmol/L) and cyclosporin A (0.08 mol/L) inhibited TF expression when added concurrently with LPS or TNF, but not when added 4 h after stimulation. TF mRNA was not detected in unstimulated alveolar macrophages by Northern blot analysis. In contrast, exposure to LPS or TNF for 6 h induced marked expression of TF mRNA, which was inhibited by treatment with pentoxifylline, retinoic acid and cyclosporin A. Expression of TNF by alveolar macrophages stimulated with LPS was also inhibited by these compounds. Our results indicate that procoagulant activity expressed by alveolar macrophages and monocytes is associated with expression of TF, whereas procoagulant activity expressed by neutrophils and platelets is not. The concentrations of pentoxifylline and retinoic acid necessary for inhibition of TF expression in vitro may not be achievable in vivo owing to their toxic effects. However, the in vitro concentration of cyclosporin A that inhibited TF expression did not exceed the plasma concentration observed in humans, and therefore may be useful for inhibition of TF expression in vivo.Abbreviations BAL bronchoalveolar lavage - LPS lipopolysaccharide - cDNA cloned deoxyribonucleic acid - cAMP cyclic adenosine monophosphate - GAPDH glyceraldehyde phosphate dehydrogenase - mRNA messenger ribonucleic acid - TF tissue factor - TNF tumour necrosis factor - DPBS Dulbecco's phosphate-buffered saline     

The use of cultured hepatocytes from goats and cattle to investigate xenobiotic oxidative metabolism

The oxidative metabolism of aldicarb (ALD), a carbamate pesticide, and fenbendazole (FBZ), an anthelmintic, was studied using cultured hepatocytes obtained from 4 goats and a bullock and incubated with ALD (50 mol/L) and FBZ (10 mol/L). The parent compounds and the metabolites were measured by HPLC. Both compounds are metabolized at the sulphur atom via two sequential oxidations, first to the sulphoxide (aldicarb sulphoxide and oxfendazole, respectively) and then to the sulphone. Oxfendazole and fenbendazole sulphone from FBZ, and aldicarb sulphoxide from ALD were found in both species. Aldicarb sulphone was not produced by the hepatocyte preparations from the bullock. The good correlation obtained comparing the in vitro results of FBZ metabolism with published in vivo dat obtained on FBZ kinetics in the same species confirmed the usefulness of in vitro models for predictive analysis of in vivo xenobiotic biotransformations.Abbreviations ALD aldicarb - ALDSON aldicarb sulphone - ALDSOX aldicarb sulphoxide - BSA bovine serum albumin - ID internal diameter - EGTA ethylene glycol bis(-aminoethyl ether) N,N,N,N-tetraacetic acid - FBZ fenbendazole - FBZSON fenbendazole sulphone - HBSS Hanks' balanced saline solution - HPLC high-pressure liquid chromatography - LDH lactate dehydrogenase - MFO mixed function oxidase - NCS newborn calf serum - OXF oxfendazole - WME Williams' Medium E     

Quantitative capillary reversed passive latex agglutination test for C-reactive protein (CRP) in the dog

A capillary reversed passive latex agglutination test (capillary RPLA) was developed which allows quantification of serum C-reactive protein (CRP) within approximately 15 min. The logarithmic regression line (calibration curve) obtained after measuring each CRP concentration three times in twofold dilutions of a standard canine serum containing 222 g/ml of CRP was y=6.394+0.030x (r=0.995). Capillary RPLA permitted quantification of CRP in the range 6.9–222 g/ml. The coefficients of variation ranged from 10.28% to 12.40%. The recovery rates (percentage recovery) of CRP by capillary RPLA were within the range 87% to 106%. On measuring the CRP concentrations in sera from 78 dogs by capillary RPLA, single radial immunodiffusion (SRID) and enzyme-linked immunosorbent assay (ELISA), close correlations were demonstrated between SRID and capillary RPLA (y=7.250+1.109x, r=0.978), between SRID and ELISA (y=3.042+1.059x, r=0.967), and between capillary RPLA and ELISA (y=1.778+0.929x, r=0.962). Capillary RPLA may be considered useful as a routine biochemical technique for measurement of serum CRP concentration in the dog.Abbreviations CRP C-reactive protein - ELISA enzyme-linked immunosorbent assay - RPLA reversed passive latex agglutination test - SRID single radial immunodiffusion     

The antimicrobial susceptibility of Staphylococcus species isolated from canine dermatitis

In a retrospective study, 1538 strains of -haemolysin-producing Staphylococcus species isolated from dermatitis in dogs at three veterinary clinical microbiology laboratories in Norway during 1986–87 and 1993–94 were investigated for their antimicrobial susceptibility. None of the strains was resistant to cloxacillin, cephalexin or the quinolones enrofloxacin and ciprofloxacin. More than 96% of the strains were susceptible to trimethoprim-sulphonamide, bacitracin and fucidic acid. Between 67% and 89% of the strains were susceptible to erythromycin, lincosamides, tetracycline, neomycin and chloramphenicol. Only 37.9% of the strains were susceptible to penicillin. The frequency of penicillin resistance increased significantly between the first and second periods, from 46.0% to 58.6%. The frequency of resistance to lincomycin, clindamycin and erythromycin also increased significantly between the first and second periods, from 3.0%, 2.1% and 3.3% to 25.5%, 19.5% and 24.8%, respectively. A moderate increase in resistance to tetracycline was also noted, from 20.4% in the first to 27.6% in the second period. On the other hand, the frequency of resistance to trimethoprim-sulphonamide decreased significantly from 4.1% in the first to 0.9% in the second period. Many different resistance patterns were observed in each period. However, the proportion of multiresistant strains increased from 2.1% in the first to 10.2% in the second period. There was a decrease in resistance to the combination of trimethoprim-sulphonamide and penicillin from the first to the second period. Resistance to the combination of lincosamides and pencillin increased. For the combinations penicillin-tetracycline-lincosamides, pencillin-lincosamides-erythromycin, and pencillin-tetracycline-lincosamides-erythromycin, there was a striking increase in resistance between the first and the second periods.Abbreviations CVL Central Veterinary Laboratory - NCVM Norwegian College of Veterinary Medicine - NVL Norwegian Veterinary Laboratory     

Book reviews

Manipulation of the Avian Genome. Edited by R. J. Etches & A. M. V. Gibbins, 1993. 322 pp., illustrated in colour. Boca Raton, FL, CRG Press. ISBN 0–8493–4216–3.

Proceedings of the 8th European Symposium on Poultry Nutrition. Edited by The Organizing Committee, 1992. 373 pp. Milan, Italy. World's Poultry Science Association.     

Effects of cultivar,pelleting and enzyme addition on nutritive value of barley in poultry diets

1. The effect of pelleting process and Trichoderma viride enzymes (TVE) addition on apparent metabolisable energy, corrected for nitrogen balance (AMEn) and on productive value of practical diets containing 40 and 45% of three different barley cultivars and one wheat were studied in poultry.

2. The effect of the pelleting process on AMEn was inconsistent and was dependent on the cereal included and the addition of enzyme.

3. The growth trial showed a significant effect of enzyme addition to pelleted diets over the whole growth period (0 to 42 d). Addition of TVE improved weight gain and food efficiency by 1.3% and 2.9%, respectively and decreased food intake by 1.6% between 0 and 22 d. In the finisher period (23 to 42 d) TVE improved efficiency by 2.8% and reduced food intake by 2.9%.

4. The incidence of sticky droppings was related to the viscosity of barley used, and enzyme supplementation reduced it. Both pelleting and enzyme addition increased dry matter content of excreta.

5. At the end of the experiment, 14 animals per treatment were slaughtered and carcass yield, viscera weight and abdominal fat were determined.     

Variation in plasma cholesterol concentration over time in the domestic fowl

1. Variations in the concentration of plasma cholesterol available to the developing oocyte over a three week period in Gallus domesticus are described.

2. There are small changes in concentration between weeks for individual birds, but no changes between consecutive days within weeks or times within days.

3. It is recommended that future attempts to assess the relationship between variation in blood and yolk cholesterol concentrations estimate blood cholesterol concentration from two samples taken a week apart.     

Comparative responses of genetically lean and fat chickens to lysine,arginine and non‐essential amino acid supply. I. growth and body composition

1. Three experiments were performed to study the effects of amino acid imbalance on the growth of genetically lean (LL) or fat (FL) male chickens from 28 to 42 d of age. In experiment 1, five concentrations of digestible lysine were compared (4.75, 6.75, 7.75, 8.75 and 9.75 g/kg). In experiment 2, four concentrations of digestible arginine were compared (6.53, 7.69, 8.84 and 10.0 g/kg). In experiment 3, three diets were compared: a high‐protein diet (189 g CP/kg), a low‐protein diet containing added essential amino acids (EAA) (144 g CP/kg) and this low‐protein diet supplemented with 40 g/kg of non‐essential amino acids (NEAA) (glutamic acid + aspartic acid).

2. LL birds exhibited a lower growth rate than the FL when the diet was deficient in either lysine or arginine. Plotting weight gain against lysine or arginine intake suggested that most of this effect was caused by variations in food intake.

3. When protein gains (body or total proteins) were plotted against lysine or arginine intake, LL chickens appeared more efficient than FL chickens.

4. Similar growth rates, although slightly lower in FL, were obtained with low‐ and high‐protein diets. However, NEAA supplementation of the low‐protein diet reduced adiposity of LL and did not modify that of FL. Increasing crude protein content (all amino acids) was more effective than NEAA supplementation in decreasing the adiposity of both lines.