Growth performance, duodenal morphology and the caecal microbial population in female broiler chickens fed glycine-fortified low protein diets under heat stress conditions.

1. This study was undertaken to examine the effect of feeding glycine (Gly)-fortified low protein (LP) diets on the growth performance, duodenal morphology and caecal microbial populations of broiler chickens raised under unheated, cyclic or constant heat stress environmental conditions. 2. From d 1 to 21 (starter phase), an equivalent number of birds were fed either a normal protein (NP) diet or a LP diet fortified with Gly. From d 22 to 42 (grower phase), an equivalent number of birds from each starter diet were distributed to one of the following dietary groups: (i) an NP diet during the starter and grower phases (NPNP), (ii) an NP diet during the starter phase and a LP diet during the grower phase (NPLP), (iii) an LP diet during the starter phase and an NP diet during the grower phase (LPNP) or (iv) LP diets during both phases (LPLP). 3. Commencing from d 22, an equivalent number of birds from each dietary group were exposed to (i) 23 ± 1°C throughout (unheated), (ii) 34 ± 1°C for 7 h each day from 10:00 to 17:00 (cyclic heat) or (iii) 34 ± 1°C throughout (constant heat). 4. Feeding the LP diet during the starter phase resulted in feed intake (FI), weight gain (WG), feed conversion ratios (FCR) and energy efficiency ratios (EER) similar to those for the NP diet. The birds fed the LP diet had a significantly higher protein efficiency ratio (PER) compared with the birds fed the NP diet. 5. During the grower phase, there were significant diet × temperature interactions for F, WG, FCR, PER, EER, villus height, crypt depth and caecal Clostridia. The birds fed the NPLP and LPLP diets had lower FI, WG and EER, higher FCR, shorter villus height and crypt depth and higher caecal Clostridia compared with the birds fed LPNP and NPNP diets under constant heat stress. However, feeding birds the NPLP and LPLP diets resulted in FI, WG, EER, FCR, morphology parameters and caecal Clostridia equivalent to the birds fed LPNP and NPNP diets, as well as improved PER, under unheated and cyclic heat stress conditions. 6. In conclusion, our results indicate that Gly-fortified LP diets can be fed to broilers under normal and acute heat stress environmental conditions without any adverse effects on performance. However, the use of such LP diets can be detrimental to broilers under chronic heat stress conditions.

Determination of aflatoxin M1 using a dialysis-based immunoaffinity sample pretreatment system coupled on-line to liquid chromatography. Reusable immunoaffinity columns.

A liquid chromatographic column-switching system containing a dialysis unit and an anti-aflatoxin immunoaffinity precolumn (immuno precolumn) is described for the automated determination of aflatoxin M1 in milk samples. Both a flat membrane dialysis unit working according to the flowing donor-flowing acceptor principle and a laboratory made hollow-fibre dialysis unit working according to the stagnant donor-flowing acceptor principle were evaluated. The hollow-fibre unit is superior with respect to repeatability (3% relative standard deviation) and detection limit (10 ng/l for aflatoxin M1 in milk), in spite of the fact that the overall recovery is only 6%. Interfering compounds, which would destroy the activity of the immuno precolumn, are efficiently removed from the system by the dialysis step; a single immuno precolumn can then be used for over 70 milk analyses. No decrease in the performance of either the immuno precolumn or the hollow-fibre dialysis unit is observed.

Direct introduction of large-volume urine samples into an on-line immunoaffinity sample pretreatment-capillary gas chromatography system.

An immunoaffinity precolumn containing immobilized antibodies raised against the synthetic steroid hormone beta-19-nortestosterone, has been used for the automated sample pretreatment of urine samples containing beta-19-nortestosterone or the related steroids norethindrone and norgestrel. The sample pretreatment system was coupled on-line to a capillary GC. The on-line connection between the immunoaffinity precolumn and the capillary GC was realized with an interface that consisted of a 10 mm X 2 mm i.d. reversed-phase pre-column and a diphenyltetramethyldisilazane-deactivated GC retention gap. After preconcentration on the immunoaffinity precolumn the analytes were eluted and reconcentrated on the reversed-phase precolumn. Subsequently, this precolumn was desorbed with 75 microL of ethyl acetate, which was directly introduced into the retention gap by using partially concurrent solvent evaporation. The system allows the automated pretreatment and GC analysis of 5-25-mL urine samples for the ppt-level determination of 19-norsteroids. The general applicability and potential of on-line immunoaffinity-capillary GC systems are discussed.

On-line immunoaffinity sample pre-treatment for column liquid chromatography: evaluation of desorption techniques and operating conditions using an anti-estrogen immuno-precolumn as a model system.

An immunoaffinity precolumn (immuno-precolumn) containing an immobilized antibody directed against estrogen steroids, was used as a model system for the evaluation of different desorbing techniques, suitable for on-line coupling to column liquid chromatography (LC). Desorption of estrogen analytes from the immuno-precolumn proved to be impossible with the buffers and chaotropic solutions frequently used in affinity desorption. Micellar solutions are effective in obstructing the antibody-antigen reaction, but their use as desorbing solutions was not found to be practical because of the large interferences introduced into the chromatograms. Desorption with aqueous solutions at elevated temperature, created by microwave action or conventional heating, although effective is not practical in this instance, because the agarose used in this study as the stationary phase for the immuno-precolumn is prone to heat decomposition. The most effective and practical approach is desorption with a methanol-water mixture (95 + 5 v/v). On-line dilution of the eluate allows the concentration of the desorbed analytes using a reversed-phase LC system with subsequent separation and ultraviolet detection. The performance of the system with spiked urine and plasma samples, which were introduced directly into the system, was evaluated and the results were compared with immunoselective desorption.

Determination of beta-19-nortestosterone and its metabolite alpha-19-nortestosterone in biological samples at the sub parts per billion level by high-performance liquid chromatography with on-line immunoaffinity sample pretreatment.

An immunoaffinity precolumn (immuno precolumn) packed with Sepharose-immobilized polyclonal antibodies against the anabolic hormone 17 beta-19-nortestosterone (beta-19-NT) was used for the selective on-line pretreatment of raw extracts of urine, bile and tissue samples by high-performance liquid chromatography. Using UV detection (247 nm), beta-19-NT and its metabolite 17 alpha-19-nortestosterone (alpha-19-NT) can be determined in biological samples with a detection limit of 0.05 microgram/kg. Owing to the high clean-up efficiency of the immuno precolumn and the large sample volumes used, confirmation by gas chromatography-mass spectrometry is possible at this level. In urine samples from a calf treated with 19-nortestosterone 17 beta-laurate, the maximum concentrations of beta-19-NT (1.3 micrograms/l) and alpha-19-NT (3.1 micrograms/l) were found seven days after intramuscular administration. In a bile sample from this calf only alpha-19-NT (55 microgram/l) was detected. In meat samples from three treated calves, the concentration of beta-19-NT varied from 0.1 to 1.6 micrograms/kg and no alpha-19-NT could be detected. In liver samples from these calves, the concentrations of beta-19-NT and alpha-19-NT were less than 0.05-0.1 and 0.5-0.9 micrograms/kg, respectively. In the corresponding kidney samples, the concentrations of beta-19-NT and alpha-19-NT were 0.4-0.5 and 0.5-1.6 micrograms/kg, respectively. The application of the same immuno precolumn to the determination of 17 beta- and 17 alpha-trenbolone, two structurally related steroids, is also demonstrated.

Effects of the alpha 2-adrenoceptor antagonists yohimbine and idazoxan on kidney function in intact and diabetes insipidus rats.

The purpose of the present study was to examine the effect of the alpha 2-adrenoceptor antagonists yohimbine and idazoxan on kidney function in intact Sprague-Dawley rats and in Brattleboro rats with familial hypothalamic diabetes insipidus. Both drugs, injected intravenously at 0.5 mg/kg, led to a marked decrease of urine flow, sodium, potassium and chloride excretion in both strains of rats. Glomerular filtration rate and effective renal plasma flow did not change significantly in either strain. The study demonstrates that the alpha 2-adrenoceptor antagonists induce an antidiuretic effect without changes in renal hemodynamics. Antidiuretic hormone does not play a role in yohimbine- or idazoxan-induced antidiuresis. The data suggest a role for alpha 2-adrenoceptors in the renal tubular handling of water and salt.

Immunoaffinity pre-column for selective on-line sample pre-treatment in high-performance liquid chromatography determination of 19-nortestosterone.

A liquid chromatographic column-switching system for automated sample pretreatment and determination of the anabolic hormone beta-19-nortestosterone (beta-19-NT) and its metabolite alpha-19-nortestosterone (19-norepitestosterone) in calf urine is described. The system consists of an immunoaffinity pre-column (immuno pre-column) packed with Sepharose-immobilized polyclonal antibodies against beta-19-NT, a second pre-column packed with C18 bonded silica and an analytical C18 column. Urine (25 ml) is directly loaded on the immuno pre-column, where the analytes of interest are trapped by the immobilized antibodies. Next the analytes are desorbed selectively with a solution containing an excess of the cross-reacting steroid hormone norgestrel and transferred, via the second pre-column, to the analytical column. The recovery of beta-19-NT in spiked urine samples was over 95%. The detection limit was 50 ng/l for a 25-ml urine injection. The system showed no loss of analytical performance over a 6-month period, during which about 100 samples were analysed with the same immuno pre-column. The general applicability of this sample pretreatment method is discussed.

Effect of inhibitors of prostaglandin synthesis on the furosemide action in the loop Henle of rat kidney.

Inhibitors of prostaglandin synthesis such as indomethacin and meclofenamate may attenuate the diuretic response to furosemide. The purpose of the present study was to clarify whether an inhibition of furosemide's action in the loop of Henle is involved in this interaction. In a first set of experiments, the effect of indomethacin and meclofenamate on the diuretic response to furosemide was re-evaluated in anaesthetized rats. Single loops of Henle of rat kidneys were perfused in vivo in a second group of rats. Furosemide was added to the perfusion fluid and the effect of indomethacin (5 mg/kg i.v.) and meclofenamate (5 mg/kg i.v.) on furosemide's action in the loops was tested. In the absence of furosemide, indomethacin and meclofenamate did not significantly affect urine flow and sodium chloride excretion, or the loop's sodium and chloride reabsorption. Both prostaglandin inhibitors, however, significantly attenuated the diuretic effect of furosemide (1 mg/kg i.v.) and the inhibitory action of the diuretic, at a concentration of 10(-4) M, on the loop's sodium and chloride transport. The results identify the loop of Henle as a tubular site of interaction between furosemide and prostaglandin inhibitors. The interaction may be related to a furosemide-induced stimulation of renal prostaglandins.