Reactive oxygen species (ROS) may be involved in the toxicity of chlorpyrifos-ethyl (CE) [O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl)phosphorothioate]. We have, therefore, examined the in vivo effects of CE on the rat erythrocyte antioxidant system and evaluated the ameliorating effects of melatonin and a combination of vitamin E and vitamin C on the oxidative damage induced by CE. The experimental groups were: (1) control group, (2) CE-treated group (CE), (3) vitamin E plus vitamin C treatment group (Vit), (4) melatonin-treated group (Mel), (5) vitamin E plus vitamin C plus CE treatment group (Vit + CE), and (6) melatonin plus CE treatment group (Mel + CE). Vitamin E and vitamin C were administered intramuscularly once a day for 6 consecutive days at 150 and 200 mg/kg, respectively, in the Vit and Vit + CE groups. Melatonin was administered intramuscularly at 10 mg/kg per day for 6 consecutive days in the Mel and Mel + CE groups. At the end of the fifth day, the rats of CE, Vit + CE and Mel + CE groups were treated orally with the first of two equal doses of 41 mg/kg CE, the second oral dose being given 21 h later. Blood samples were taken 24 h after the first CE administration. Levels of thiobarbituric acid reactive substance (TBARS), antioxidant defence potential (AOP), and the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) were determined in erythrocytes. In comparison with the control group, oral administration of CE significantly (P < 0.05) stimulated TBARS activity while significantly (P < 0.05) inhibiting AOP and the activities of SOD and CAT. However, GSH-Px activity remained unchanged by CE treatment. Treatment with melatonin and vitamins E plus C significantly (P < 0.05) reduced the CE-induced increase of TBARS, and overcame the inhibitory effect of CE on SOD and CAT, but not on AOP. Melatonin treatment significantly (P < 0.05) increased only GSH-Px activity, irrespective of the effect of CE. These results suggest that CE treatment increases in vivo lipid peroxidation and decreases antioxidant defence by increasing oxidative stress in erythrocytes of rats, and melatonin and a combination of vitamin E and vitamin C can reduce this lipoperoxidative effect.

1. Two experiments were carried out to investigate the addition of 1 3 kg water per kg air-dry mash diets containing high proportions (600 to 700 g/kg) of ground cereal grains (wheat, barley or oats) on broiler performance and the structure and function of the gastro-intestinal tract. 2. Chicks at the age of 7 d were fed on the wheat-, barley- or oats-based diets in the dry or wet forms for 35 d. Food and water intakes were recorded daily while body weight was measured weekly. Two birds from each treatment were killed each week to measure gut size and the viscosity of gut contents. Tissue samples from various digestive segments were histo-morphologically examined to determine the thickness of tissue layers, size of tissue glands, villa heights, crypt depths and thickness of tunica muscularis. Crypt cell proliferation rate (CCPR) for each segment was also determined using a metaphase arrest technique. 3. The results from both experiments showed that wetting food significantly (P<0.05) increased food intake, total water intake and body weight gain of broiler chickens. The body weight gains of birds were proportional to their food intakes so that the efficiency of food utilisation was similar for all treatments. Dry matter retention of food tended to increase in birds given wet food from 7 to 21 d but not thereafter, compared to the dry-fed birds. Although water intake from the water bottle was significantly (P<0.05) reduced in birds given wet food, total water intakes from the water bottle plus that from food were significantly (P<0.05) higher in the wet-fed birds than in the dry-fed birds. The ratio of total water to dry food intake was, however, similar in both feeding regimens. 4. The fresh empty weight of the gut was increased by wet-feeding while its relative weight to body weight and the length of gut was not affected by dietary treatments. Significantly greater development of the tissue glands in the proventriculus and gizzard was observed in the birds given wet food; this was associated with the reduced thickness of the muscular layer of these segments. An increase in villus height was also observed in duodenum, small intestine, caeca and colon of birds given wet food, compared to those given dry food. CCPR was significantly (P<0.05) reduced by wet-feeding throughout the digestive tract. This was associated with a significant decrease in the mean viscosity of the gut contents and the concentration of volatile fatty acids (VFAs) in the caeca. 5. Wetting diets based on cereal grains caused a significant improvement in the performance of broiler chickens. The mechanism of the beneficial effects of wet feeding could be attributed to the decreased viscosity of gut contents; the greater development of the layer of villi in the digestive segments and the reduced CCPR in the crypts of the epithelium.

To test whether the improvements in digestive ef®ciency due to either wetting of the food or inclusion of enzymes are accompanied by the same changes in gut function, foods with a high content of wheat were fed to broiler chicks from 1±42 d old. Twenty-four birds were caged individually while a further sixty-four were in group pens in experiments of 2´2 factorial design with two levels of enzyme (0 or 1 g/kg, designed for wheat) and two levels of water addition (0 and 1300 g/kg). Food intake and live-weight gain were signi®cantly increased by wet feeding (from 89×3 to 153×4 g/d and from 39×7 to 65×4 g/d respectively), the differences increasing with age, while the enzyme had no signi®cant effect (120×5 and 122×2 g/d and 51×9 and 53×1g/week respectively). The viscosity of digesta was greatly reduced both by wetting (from 4×40 to 2×64 kPa × s) and enzyme (from 4×47 to 2×57 kPa × s) but there was a signi®cant interaction with age in which the viscosity was low throughout in the wet only, enzyme only and wet + enzyme treatments but declined with age from a very high level in the dry, no enzyme treatment (11×5 kPa × s at 14 d). While wetting increased weight and length of digestive tract and thickness of some parts of the gut, enzyme had no signi®cant effect, tending to reduce gut wall thickness. Crypt cell proliferation rate (CCPR) was signi®cantly reduced by wet feeding (from 39×4 to 28×7 cells/crypt per 2 h) and by enzyme supplementation (from 38×9 to 29×2 cells/crypt per 2 h). Therefore, while both wetting and enzyme addition to the food reduced digesta viscosity and CCPR to a similar extent, the former had marked stimulatory effects on food intake and weight gain while the latter had little effect. The mode of action of wet feeding is therefore deduced to be not primarily through its effects on viscosity and CCPR.

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