A modified differential stain for cartilage and bone in whole mount preparations of mammalian fetuses and small vertebrates

Fixation in formol-acetic-alcohol as a prelude to the staining of whole mount vertebrate skeletons with alcian blue and alizarin red S has greatly facilitated the enzyme clearing step of the method outlined by Dingerkus and Uhler. The modified method has been tested on fetal and neonatal mice, and on a variety of vertebrates including bony fish, reptiles, amphibia and birds, and shown to be rapid, reproducible and permanent. The method is not so rapid as that reported by Kimmel and Trammell but is superior at least in certain circumstances. In the present study, optimal results were obtained by fixing in formol-acetic-alcohol for 40 minutes, staining cartilage with alcian blue 8GX, then clearing with trypsin. The time taken to complete the latter step was reduced significantly by incubation at 37 C. The next step was to stain bone using alizarin red S in a weak solution of potassium hydroxide, followed by clearing in a potassium hydroxide-glycerol series

Ileal digestibility of amino acids in feed ingredients for broilers.

To more precisely formulate feed and predict animal performance, it is important to base both the recommendations and feed formulations on digestible rather than total amino acid contents. Most published data on the digestibility of amino acids in feed ingredients for poultry are based on excreta digestibility. Ileal digestibility is an alternative and preferred approach to estimate amino acid availability in feed ingredients. Both methodologies are described and assessed. In addition, the differences between apparent and standardised (in which corrections are made for basal endogenous losses) digestible amino acid systems are discussed. The concept of a standardised digestibility system as a mean of overcoming the limitations of apparent digestibility estimates is proposed. In this context, different methodologies for the determination of basal endogenous amino acid losses are discussed. Although each methodology suffers from some limitations and published data on endogenous losses at the ileal level in growing poultry are limited, averaged data from repeated experiments using the 'enzymatically hydrolysed casein' method are considered as the best measure of basal losses. Standardised ileal amino acid digestibility values of 17 feed ingredients commonly used in broiler nutrition are presented including grains (barley, corn, sorghum, triticale, wheat), grain by-products (wheat middlings, rice pollard), plant protein sources (soybean meal, canola meal, corn gluten meal, cottonseed meal, lupins, peas/beans, sunflower meal), and animal by-products (feather meal, fish meal, meat and bone meal). This comprehensive set of the ileal amino acid digestibility of feed ingredients in broiler nutrition may serve as a basis for the establishment of the system in broiler feeding and for further research

Influence of age on the apparent ileal amino acid digestibility of feed ingredients for broiler chickens

1. The apparent ileal digestibility of amino acids in 8 feed ingredients was determined using 14-, 28- and 42-d-old male broiler chickens. The ingredients included three cereals (wheat, sorghum and maize), one cereal by-product (mill run), three oilseed meals (canola, cottonseed and soybean meals) and one animal protein meal (meat and bone meal). 2. Dietary crude protein in the assay diets was supplied solely by the test ingredient. All diets contained 20 g/kg acid-insoluble ash as an indigestible marker, and each diet was offered ad libitum in mash form to 5 replicate pens from 11 to 14 d, 25 to 28 d and 39 to 42 d post-hatching. There were 12, 6 and 6 birds per pen for the 14, 28 and 42 d samplings, respectively. 3. The results suggest that the age of broilers significantly influenced the apparent ileal digestibility of amino acids. The effects, however, varied among amino acids and ingredients. 4. In wheat, the digestibility of most amino acids was higher in 14- than in 28- and 42-d-old broilers. In maize, the digestibility coefficients of amino acids were higher at 28 and 42 d than at 14 d. The digestibility coefficients in maize and wheat at 28 and 42 d were similar. The digestibility of amino acids in sorghum at 42 d was higher than those at 28 d, but similar to those at 14 d except for histidine, lysine, serine and glycine, which were significantly higher at 42 d. Digestibility of amino acids in sorghum was similar between 14 and 28 d except for isoleucine, leucine, phenylalanine, glutamic acid and alanine. The digestibility of amino acids in mill run at 42 d was significantly higher than those at 14 and 28 d. There were no differences in digestibility between 14 and 28 d. 5. In general, the digestibility of amino acids in canola meal, soybean meal and, meat and bone meal was higher at 28 and 42 d compared to those at 14 d, and similar between 28 and 42 d of age. In cottonseed meal, age had no effect on the digestibility coefficient of amino acids, except for lysine and arginine, which increased with age. 6. Analysis of the combined results for the 8 feed ingredients showed that, in general, the digestibility coefficients of amino acids increased with advancing age of broiler chickens

Nutrient utilisation by the hindlimb of Thoroughbred horses at rest

Nutrient uptake by the hindlimb was investigated utilising the arteriovenous difference technique in 5 Thoroughbred horses fed to maintenance a diet of 100% roughage or 52% oat grain and 48% roughage. Arterial blood was obtained from a catheter inserted into the carotid artery while venous blood was simultaneously collected from a catheter placed into the iliac vein via the medial saphenous vein. The arteriovenous difference for glucose was significant and represented a mean extraction of 10 ± 1% with no effect of diet. If fully oxidised, glucose uptake (corrected for lactate and pyruvate arteriovenous difference) was sufficient to account for 78 ± 13% or 107 ± 15% of the oxygen consumed by the hindlimb in horses fed a roughage or 52% oat grain diet respectively. Acetate was also a major metabolite of the hindlimb, showing a 39 ± 5% extraction with no effect of diet. However, the 52% oat grain diet did induce a significant decline in the concentration of acetate in arterial blood. The potential contribution to oxidation in the hindlimb was significantly reduced from 32 ± 4% in horses fed roughage to 21 ± 3% when fed 52% oat grain. D-3-Hydroxybutyrate uptake could account for 9 ± 1% of the oxidation by the hindlimb with no effect of diet. The technique for measuring nutrient uptake across the hindlimb using the arteriovenous difference is relatively simple and would be valuable in investigating fuel use by muscle during exercise