James L. Atkinson

Response of rainbow trout (Salmo gairdneri) to increased levels of available carbohydrate in practical trout diets.

1. The physiological response of rainbow trout (Salmo gairdneri) reared on different levels of available carbohydrate in practical trout diets having the same levels of energy and nitrogen for 16-24 weeks was determined. 2. Weight gain was significantly reduced in trout reared on the highest level of available carbohydrate, 210 g cerelose (alpha-glucose) kg, and there was a significant linear regression (R2 0.88 of dietary carbohydrate on weight gain. 3. Liver: body-weight values and liver glycogen levels increased in relation to increased dietary carbohydrate. 4. Liver glucose-6-phosphate dehydrogenase (EC 1.1.1.49) activity increased and liver phosphoenolpyruvate carboxykinase (EC 4.1.1.32) activity decreased per kg body-weight of fish with increasing dietary carbohydrate. However, no significant effect was noted on the activity of these liver enzymes above a dietary cerelose level of 140 g/kg. 5. Liver fructose diphosphatase (EC 3.1.3.11) activity increased with increasing dietary carbohydrate has been interpreted as meaning a recycling of triosephosphate to glucose-6-phosphate. 6. Dietary carbohydrate level had no significant effect on the liver pyruvate kinase (EC 2.7.1.40) activity, the rate of glucose utilization or the percentage conversion of [14C]alanine to glucose in the plasma of trout. 7. The results indicate that rainbow trout have a limited ability to adapt to increased dietary carbohydrate and a level in excess of 140 g/kg of the diet is not efficiently utilized.

Proline as an essential amino acid for the young pig

1. The catabolism of L-[1-14C]phenylalanine was used to indicate the effects of single amino acid supplementation of an inadequate protein diet (200 g crude protein (nitrogen x 6.25)/kg) on the utilization of dietary amino acids in pigs of 2.5 kg body-weight reared on an adequate protein diet (240 g crude protein/kg) containing skim milk and a mixture of free amino acids. 2. The oxidation of phenylalanine was decreased by the addition of proline or arginine to the inadequate protein diet but not by the addition of threonine, methionine, lysine or a mixture of essential amino acids, indicating that proline and arginine were limiting the utilization of dietary amino acids in the inadequate protein diet. 3. Dietary proline concentrations of 13.9 and 14.2 g/kg minimized phenylalanine oxidation in diets containing 200 or 260 g protein/kg. This indicates a dietary proline requirement of 14 g/kg. 4. Increasing the dietary arginine concentration in a diet containing 240 g protein/kg showed that an arginine concentration of 5.1 g/kg minimized phenylalanine oxidation. However, increasing the arginine concentration in a diet containing 200 g protein/kg increased phenylalanine oxidation, suggesting an amino acid imbalance involving arginine at this lower level of protein.

Effect of dietary tryptophan on plasma and brain tryptophan, brain serotonin, and brain 5-hydroxyindoleacetic acid in rainbow trout

In order to determine the effect of dietary tryptophan level on plasma and brain tryptophan, brain serotonin, and brain 5-hydroxyindoleacetic acid levels, juvenile rainbow trout (Salmo gairdneri) were raised for 16 weeks on semipurified diets containing 0.06%, 0.16%, 0.21%, 0.26%, 0.39%, or 0.59% tryptophan. After 14 weeks, feed intake was depressed in fish fed the diets containing 0.06% or 0.16% tryptophan. No further differences in feed intake were noted between the remaining treatments. In addition, body weight was lower in fish fed diets containing 0.06%, 0.16%, or 0.21% tryptophan compared with fish fed higher levels. After 16 weeks of feeding the test diets, plasma tryptophan levels were found to be directly related to dietary tryptophan levels. Similarly, increased dietary levels of tryptophan resulted in increased brain levels of tryptophan, serotonin, and 5-hydroxyindoleacetic acid. These results demonstrate that in rainbow trout, as in mammals, altered dietary levels of tryptophan result in alterations in plasma and brain tryptophan, brain serotonin, and brain 5-hydroxyindoleacetic acid.

Pollen substitutes increase honey bee haemolymph protein levels as much as or more than does pollen.

Summary Adequate substitutes for pollen are necessary for maintaining healthy bee colonies during periods of pollen dearth, but testing them objectively is both time consuming and expensive. We compared two commercial diets with bee collected pollen and acacia pod flour (used by beekeepers in some parts of Brazil) by measuring their effect on haemolymph protein contents of young bees exclusively fed on these diets, which is a fast and inexpensive assay. The commercial diets included a new, non-soy-based, pollen substitute diet (named Feed-Bee®) and a soy-based diet, named Bee-Pro®. The diets were each given in patty form to groups of 100 Africanized honey bees in hoarding cages, maintained and fed from emergence until six days of age. Sucrose, in the form of sugar syrup, was used as a protein free control. Feed-Bee®, Bee-Pro®, pollen and acacia pod flour diets increased protein titers in the haemolymph by factors of 2.65, 2.51, 1.76 and 1.69, respectively, over protein titers in bees fed only sucrose solution. The bees fed Feed-Bee® and Bee-Pro® had their haemolymph significantly enriched in protein compared to the controls and those fed acacia pod flour and to titers slightly higher than those fed pollen. All four proteinaceous diets were significantly superior to sucrose alone.

Evaluation of the net energy value of glucose (cerelose) and maize starch in diets for rainbow trout ( Salmo gairdneri )

1. Quadruplicate groups of rainbow trout (Salmo gairdneri) (mean body-weight 24.9 g) were reared on six dietary treatments (practical-type diets) in a modified paired-feeding experiment for 12 weeks at 15 degrees to determine the net energy (NE) value of starch and glucose to rainbow trout. 2. Three test diets were prepared to contain (g/kg): 0 supplemented carbohydrate (diet 1), 250 maize starch (diet 2) and 250 glucose (diet 3) and were given ad lib. to the trout with the feeding rate of the glucose- and starch-fed groups being monitored after each feeding. The remaining three treatments involved controlled feeding of the trout with diet 1 at 75% of the feed intake of trout reared on diets 2 and 3, so as to provide the same levels of protein and lipids without carbohydrate, and with diet 2 at 100% of the feed intake of trout reared on diet 3. 3. The difference in the final carcass energy of the ad lib.-fed group and the respective controlled-fed group divided by the amount of dietary glucose or starch energy consumed by the trout is the NE value for that carbohydrate. 4. The determined NE value of glucose was 3.99 kJ/g and starch 2.17 kJ/g, which is 24.6 and 12.6% respectively of the gross energy values of these carbohydrates in rainbow trout. 5. The results indicate that digestible energy and calculated metabolizable energy values for carbohydrates in rainbow trout overestimate the utilizable energy content of the diet.(ABSTRACT TRUNCATED AT 250 WORDS)

Development and validation of a behavioral acclimation protocol for cats to respiration chambers used for indirect calorimetry studies.

Cats exposed to novel environments initiate stress responses by behavioral and physiological changes that modify metabolism and lead to the collection of unreliable data. Fourteen cats (10 ± 2 months) were subjected to an 11-week acclimation procedure to adapt to restriction within chambers used for indirect calorimetry studies. Cats were acclimated to chambers in their home environments, to chambers in the study room, and to increasing periods of restriction within chambers. Ten additional cats (11 ± 1 month), used as controls, were subjected to a single 5-hr restriction without any prior exposure. Stress level, feed intake, fearfulness, and eliminations were recorded. Latencies to approach a novel object peaked on Weeks 4 and 8 (p < .05). Cat-Stress-Scores (CSS) declined with exposure and on Week 11, stress levels were low and consistent (p < .05). CSS was greater in unacclimated versus acclimated cats (p < .05). In conclusion, acclimation protocols prepare cats for repeated, temporary restriction within chambers, whereas short acclimations do not. A step-up acclimation procedure with behavioral indices of stress should be utilized to prepare cats for research that necessitates restriction.

Effect of PCPA or tryptophan on brain serotonin and on consumption of a high protein or high carbohydrate diet by rainbow trout, Oncorhynchus mykiss

Abstract The effect of increased or decreased brain serotonin on consumption of a high protein or a high carbohydrate diet by rainbow trout, Oncorhynchus mykiss , was evaluated. Groups of rainbow trout were acclimated to either a high protein or a high carbohydrate diet for 6 weeks. Fish then received a dietary pretreatment meal containing 5% p-chlorophenylalanine, which decreased brain 5-HT and 5-HIAA, or 7% crystalline tryptophan, which increased brain 5-HIAA and 5-HIAA/5-HT. Intake of a challenge meal (high protein or high carbohydrate, consistent with the diet to which they had been acclimated) was then determined. Food consumption by fish fed the high protein diet was unaffected by either dietary pretreatment meal. Conversely, both p-chlorophenylalanine and tryptophan pretreatment decreased food consumption by fish fed the high carbohydrate diet. These results suggest that changes in brain serotonin turnover in rainbow trout are not important in controlling intake of a high protein diet. Furthermore, because consumption of the high carbohydrate diet was decreased both by p-chlorophenylalanine, which decreased serotonin turnover, and by tryptophan, which increased serotonin turnover, this also suggests that serotonin turnover may not be an important signal controlling consumption of high carbohydrate diets by rainbow trout.

Oral and intravenous l-[1-13 C]phenylalanine delivery measure similar rates of elimination when gastric emptying and splanchnic extraction are accounted for in adult mixed hounds.

There are few reported estimates of amino acid (AA) kinetics in adult mammals and none exist in adult dogs. The study objectives were to evaluate the use of oral isotope delivery in contrast to the more commonly used intravenous (IV) delivery to estimate AA kinetics in adult dogs and to estimate splanchnic extraction and gastric emptying using a commonly accepted mathematical model. Dogs received 25 × 1/2-hourly meals (13 g/kg BW/day) and either an oral or IV bolus of l-[1-(13) C]Phe (12 mg/kg BW). Blood samples were taken immediately before each feeding. Concentrations of plasma Phe were measured using liquid chromatography-tandem mass spectrometry. There were no differences in baseline plasma Phe concentrations (34 μm ± 0.61), Phe distribution volume, Phe pool size and rate constants between dogs when the tracer was administered IV or orally (p > 0.25). Decay curve for plasma l-[1-(13) C]Phe differed between IV and oral dosing protocols with IV dosing fit best using a two-compartment model. Phe disappeared from plasma at a mean rate of 2.8%/min. Estimates of gastric emptying and splanchnic extraction did not differ based on oral or IV tracer dosing when the decay curves were fit with the two-compartment model (p > 0.40). The half-life for gastric emptying was 18 min, and first-pass Phe extraction by the splanchnic bed was 24% of the dietary Phe. These results suggest that oral isotope dosing can be used as an alternative to IV isotope dosing in studies that utilize a primed, constant dosing approach to measure protein and amino acid kinetics.

Whole-body retention of α-linolenic acid and its apparent conversion to other n -3 PUFA in growing pigs are reduced with the duration of feeding α-linolenic acid

In the present study, fifteen growing pigs were used to determine the whole-body oxidation, retention efficiency (RE) and apparent conversion (AC) of α-linolenic acid (18 : 3n-3) to n-3 highly unsaturated fatty acids (HUFA), including EPA (20 : 5n-3) and DHA (22 : 6n-3). The pigs were fed a diet containing 10% flaxseed for 30 d. Whole-body fatty acid composition was determined at initial (27.7 (SE 1.9) kg), intermediate (day 15; 39.2 (SE 1.4) kg) and final (45.7 (SE 2.2) kg) body weight. On day 12, four pigs were fed 10 mg/kg of uniformly labelled (13)C-18 : 3n-3 (single-bolus dose) to determine the oxidation of 18 : 3n-3. Expired CO2 samples were collected for 24 h thereafter. The whole-body content of n-3 PUFA increased linearly (P< 0.0001) with time; however, the content of 22 : 6n-3 exhibited a quadratic response (P< 0.01) with a peak occurring at 15 h. As a proportion of intake, the RE of 18 : 3n-3 tended to reduce with time (P = 0.098). The AC of ingested 18 : 3n-3 to the sum of n-3 HUFA was reduced with time (P< 0.05; 12.2 v. 7.53 % for days 0-15 and days 15-30, respectively). The AC of 18 : 3n-3 to 20 : 5n-3 or 22 : 6n-3 was lower than that to 20 : 3n-3, both for days 0-15 (P < 0.05; 1.14 or 1.07 v. 7.06 %) and for days 15-30 (P< 0.05; 1.51 or 0.33 v. 4.29 %). The direct oxidation of 18 : 3n-3 was 7.91 (SE 0.98) % and was similar to the calculated disappearance of 18 : 3n-3 between days 0 and 30 (8.81 (SE 5.24) %). The oxidation of 18 : 3n-3 was much lower than that reported in other species. The AC of 18 : 3n-3 to n-3 HUFA was reduced over time and that to 20 : 3n-3 in the present study was much higher than that reported in other species and should be explored further.

Calibration and validation of a carbon oxidation system and determination of the bicarbonate retention factor and the dietary phenylalanine requirement, in the presence of excess tyrosine, of adult, female, mixed-breed dogs1,2,3

Carbon oxidation methods have been used as rapid and sensitive methods to determine whole-body AA requirements in multiple species. The objectives of the current studies were to validate complete CO recovery, determine the bicarbonate retention factor, and estimate the Phe requirement, in the presence of excess Tyr, in adult dogs using the direct oxidation technique. In this series of studies, 2 oxidation chambers were constructed and calibrated to ensure accurate collection of breath CO. First, 104.6 ± 7.1% CO was recovered from chambers and suggests that the chambers were appropriately designed for complete and efficient CO recovery. Second, we determined bicarbonate retention in 5 dogs using repeated oral dosing of a bicarbonate tracer (NaHCO) with small meals. At isotopic and physiological steady state, 102.5 ± 2.6% of the delivered NaHCO was recovered in breath. Third, the Phe requirement, when Tyr was supplied in excess, was determined by the rate of appearance of CO in the breath (CO). Dogs ( = 5) were fed test diets with different concentrations of Phe ranging from deficient to excessive for 2 d prior to conducting the tracer studies. The mean Phe requirement (when Tyr was supplied in excess) was 0.535% of diet (upper 95% confidence interval = 0.645% diet) on an as-fed basis or 0.575% of diet (upper 95% confidence interval = 0.694% of diet) on a DM basis and was based on a calculated (modified Atwater calculation) dietary ME density of 3.73 Mcal/kg DM. These data support the use of carbon oxidation methods and oral dosing of isotope to measure whole-body requirements of indispensable AA in adult dogs and suggest the current recommendations may be low.