Purification and characterisation of chicken liver monomeric glutathione peroxidase

1. A novel glutathione peroxidase, which is distinct from tetrameric glutathione peroxidase, was purified to homogeneity from a broiler chick liver cytosolic fraction using 5 different column chromatographic methods.

2. The enzyme in cytosol was separated from ‘classic’ tetrameric glutathione peroxidase and glutathione S‐transferases by DEAE‐Sephacel and Sephadex G‐100 chromatographies and further purified by Mono Q, hydroxylapatite and sulphobro‐mophthalein‐S‐glutathione‐agarose chromatographies.

3. The molecular weight of the purified enzyme determined by sodium dodecyl sulphate‐polyacrylamide gel electrophoresis was 19,500 and that found by gel filtration chromatography was comparable. This indicates that the enzyme protein is a single polypeptide. The isoelectric point of the enzyme was determined as 7.0 by polyacrylamide gel isoelectric focusing.

4. The purified enzyme catalysed the reduction of hydrogen peroxide, cumene hydroperoxide, tert‐butyl hydroperoxide and linoleic acid hydroperoxide. Furthermore, it reduced phosphatidylcholine hydroperoxide in the absence of phospholi‐pase A2. The optimum pH for the enzyme reaction was 7.0. The antiserum against the purified enzyme reacted with the 19.5 kDa polypeptide in the liver cytosol of duck and quail.     

Performance of broiler chicks fed on alkali‐treated neem (Azadirachta indica) kernel cake as a protein supplement

1. An experiment was conducted to evaluate growth and nutrient efficiency of broiler chicks from 3 to 42 d fed on diets containing alkali‐treated neem (Azadirachta indica A. juss) kernel cake (NKC) as a protein supplement in place of peanut meal (PNM).

2. NKC was treated with sodium hydroxide at 10 (ANKC 1) or 20 g (ANKC 2)/kg and incorporated into the test diets at 135 or 300 g/kg to replace 50 (low—L) or 100 (High—H) % of the PNM protein of the reference diet.

3. Despite comparable retentions of dry matter and total carbohydrate on L‐ANKC 1 and 2, fibre on L‐and H‐ANKC 2 and nitrogen, calcium and acid detergent fibre on all experimental diets compared to the retentions of chicks on the reference diet, only the chicks fed L‐ANKG 2 were found to grow and utilise food as well as those on the reference diet.

4. The activities of serum alkaline phosphatase on H‐ANKC 1 and alanine amino transferase on all test diets were depressed (P < 0.05), but the activity of serum aspartate amino transferase, total erythrocyte count and concentration of blood haemoglobin and urea were similar in all chicks.

5. No significant differences were noticed in the qualitative and quantitative characteristics of the meat of chicks fed on the reference diet and on diets incorporating ANKC at the lower concentrations. Feeding ANKC protein did not impart any untoward taste as evaluated in pressure cooked meat by a semitrained panel on a 7 point Hedonic scale.

6. Except for duodenal and jejunal inflammation in chicks on both reference and test diets, all the vital organs were normal, ruling out any adverse affects caused by residual neem bitters.

7. Comparable performance and cost of chicks fed on the reference and L‐ANKC 2 diets, warrants the utilisation of hitherto wasted protein‐rich NKC after alkali treatment in broiler chick diets to spare peanut meal for human consumption in developing countries.     

Changes in blood and spleen lymphocyte populations following antigen challenge in immature male chickens

1. The effects of antigen (Ag) injection on the distribution of lymphocyte populations of Cornell K‐strain male chickens were studied.

2. Two experiments were conducted. In the first, chickens were injected with Brucella abortus (BA), a purported T‐independent antigen. In the second, chickens were injected with sheep red blood cells (SRBC), a T‐dependent antigen. Peripheral blood lymphocytes (PBL) and spleen lymphocytes isolated at 0, 3, 6, 9, 12 and 24 h following Ag injection were stained with monoclonal antibodies (mAb) detecting B‐lymphocytes, CD4⁺ and CD8⁺ cells.

3. B‐lymphocytes in the blood or spleen showed no significant changes following either BA or SRBC injection. In contrast, CD4⁺ cells were decreased in the blood and increased in the spleen following BA and SRBC injections. CD8⁺ cells were decreased in both blood and spleen following BA injection but were unchanged in either blood or the spleen following SR8C injection.

4. These results indicate that there is a change in both spleen and circulating lymphocyte populations, especially T‐helper cells, following Ag injection. T‐helper cells are apparently the primary population involved in the initiation of humoral immunity.