Dietary protein effects on hen performance and nitrogen excretion.

Because dietary nitrogen intake affects nitrogen content in manure, diet management has been recognized as a means to reduce ammonia emissions from poultry operations. The objectives of the present research were 1) to determine the extent to which the CP content of laying diets can be reduced, based on performance criteria, and 2) to determine how ash:nitrogen ratios of manure, eggs, and hens are affected by dietary protein changes. Egg-type hens were fed equal daily amounts of essential amino acids in diets that provided 13, 15, or 17 g of protein/d. Each diet was fed to 20 hens, with 2 hens/cage. The planned digestible lysine intake was 0.71 g/hen per day. Ratios of other digestible amino acids to lysine were methionine plus cysteine, 0.83; threonine, 0.68; and isoleucine, 0.94. The experiment began when hens were 29 wk old and continued until they were 57 wk old. Egg production averaged approximately 90%, and daily protein intake caused no effects on egg production or grams of egg per hen per day. Feed intake was higher for hens fed 13 g of protein than for hens in the other 2 treatments (P < 0.01). Average feed intake for the experiment was approximately 95 g/d. Composition of the eggs was not affected by protein intake. Average values were DM, 30.5%; ash, 31.0% of DM; and nitrogen, 6.31% of DM. The average manure DM production was 25.9 g/hen per day, with an ash content of 25.5% of DM. Manure nitrogen content ranged from 3.98% of DM for hens fed 13 g of protein to 5.68% for those fed 17 g of protein (P < 0.01). A method is outlined that uses the analysis of fresh manure and manure leaving the poultry operation to estimate the loss of nitrogen as ammonia.

Technical note: a gene delivery system in the embryonic cells of avian species using a human adenoviral vector.

Adenovirus (Ad) has been used in vivo and in vitro as a vector to carry a foreign gene for efficient gene delivery into various cell types and tissues of animals. The aim of the current study was to evaluate the Ad delivery system in primary avian cells. Primary cells isolated from the embryonic pectoralis major muscles of the chicken and quail were cultured and incubated with human recombinant Ad serotype 5 (Ad5) containing sequences encoding either the green fluorescence protein (GFP) gene alone, as a tracking marker, or both GFP and murine 3-hydroxyisobutyryl-CoA hydrolase (mHIBCH) as a target gene. The fluorescent GFP images showed the successful delivery of a target gene using Ad5 in the primary avian cultured cells. In addition, immunostaining of the myosin heavy chain (MyHC) in these cells indicated that a large population of the cells was myogenic. Colocalization of GFP-positive cells with MyHC staining was mostly found in MyHC-negative cells, indicating successful delivery of Ad5 into a large population of mononucleated cells. Furthermore, the current fluorescence study detected the dual expression of GFP and mHIBCH protein in GFP-positive cells. Finally, Western blot analysis confirmed that the Ad-mediated expression of mHIBCH protein was specific in primary cultures of avian myogenic cells and that the mHIBCH protein expression was continued for 15 d after infection in chicken primary cells. These data demonstrate that Ad5 is a feasible tool to express foreign genes in primary cultured cells of avian species, providing a new approach to study the function of genes of interest in muscle development and metabolism.

The ontogeny of delta-like protein 1 messenger ribonucleic acid expression during muscle development and regeneration: comparison of broiler and Leghorn chickens.

Delta-like protein 1 (DLK1) has been implicated in the muscle hypertrophy observed in DLK1 transgenic mice, callipyge sheep, and mouse paternal uniparental disomy 12 and human paternal uniparental disomy 14 syndromes. The current study was aimed to determine chicken DLK1 (gDLK1) mRNA expression during primary muscle cell differentiation and during muscle regeneration after cold injury and to compare gDLK1 mRNA expression during skeletal muscle development in layers and broilers. In chicken primary muscle cell culture, gDLK1 mRNA expression was significantly increased from 12 to 48 h (P < or = 0.05) when the nascent myotubes were actively formed at d 2 to 3. Myogenin, a late myogenic marker gene, mRNA expression peaked at 36 to 48 h. Myogenic differentiation 1 (MyoD) and paired box gene 7 (Pax7), early myogenic marker genes, mRNA expression gradually decreased during myogenic differentiation. During muscle regeneration, the expression of MyoD and Pax7 peaked at d 2 (P < or = 0.05), and myogenin mRNA expression peaked at d 4 (P < or = 0.05). The induction of gDLK1 gene appeared between d 7 to 10 postinjury (P < or = 0.05) when myotubes were actively formed as also demonstrated in histological sections. The expression of gDLK1 was slowly downregulated to the control levels at d 14 when the damaged muscle appeared nearly healed. These data suggest that gDLK1 may be involved in the late myogenic stages of primary muscle cell differentiation and muscle regeneration. The gDLK1 mRNA in the muscle tissues was very abundant at embryonic ages but decreased after hatching in both broiler and layer chickens. Compared with layers, broiler muscle at embryonic d 13 had a 3-fold greater expression of DLK1 (P < or = 0.01). In addition, the gDLK1 mRNA expression at d 1, 11, and 33 post-hatch was significantly higher in broilers than layers (P < or = 0.05). Therefore, the relatively greater expression of the gDLK1 gene in muscles of broilers compared with layers suggests that gDLK1 may serve as a new selection marker for high muscle growth in chickens. These findings may provide new insight into chicken muscle development and regeneration.

Cloning of adipose triglyceride lipase complementary deoxyribonucleic acid in poultry and expression of adipose triglyceride lipase during development of adipose in chickens.

Increasing the breakdown of stored fat in adipose tissue leads to reducing fat content, enhancing feed efficiency and, consequently, decreasing the production cost of poultry. The processes of lipolysis are not completely understood, and the proteins involved in this process need to be identified. An adipose triglyceride lipase (ATGL), recently identified in several species, has not been studied in avian species. We have cloned the full-length coding sequences of ATGL cDNA for the chicken, turkey, and quail. Sequence comparisons among mammals and these avian species showed that the avian ATGL have 2 conserved domains, the patatin domain and the hydrophobic domain. The patatin domain contains lipase activity, and the hydrophobic domain exhibits lipid droplet binding. The high levels of chicken, turkey, and quail ATGL mRNA and protein are exclusively found in subcutaneous and abdominal adipose tissues. In addition, chicken ATGL (gATGL) is mainly expressed in the fractionated adipocytes compared with stromal-vascular cells that mostly contain preadipocytes (P < 0.001). Furthermore, ontogeny of gATGL mRNA and protein expression in adipose tissue showed induction of gATGL immediately after hatching before access to food (P < 0.05), suggesting that an energy deficit due to posthatching starvation may increase breakdown of stored fat via increasing gATGL expression in adipose tissue. Our studies showed that expression of the chicken ATGL is adipose specific and regulated developmentally, suggesting that a possible modulation of ATGL expression would regulate fat deposition in avian species.

The partitioning of metabolizable energy by broiler chickens.

Broiler chickens were used to alter the partitioning of ME between maintenance and production. They were fed amounts of feed that ranged between ad libitum and 75 to 80% ad libitum each day from a BW of 1.1 to 2.2 kg. The experiment was done with 2 strains of broilers, both sexes, and 2 forms of feed (mash or pellets). Data were collected to determine live performance, digestibility of the feed, and carcass composition. The daily amount of feed did not affect the ME of the feed, but broilers fed limited amounts of feed gained less BW per day and had a larger feed:gain ratio. For most measurements, strain of broiler, sex, and form of feed also were significant factors. Because broilers that gained BW more slowly required more days to gain 1.1 kg and more feed during that time, a larger proportion of the energy was used for maintenance. The net energy theory proposes that heat increment, net energy for maintenance, and net energy for production are constants for a feed ingredient or feed. Results from the present research did not support the net energy theory. A different model was proposed that used ME as the basis for energy partitioning. Amounts of feed or energy per day had no effect on the ME content of the feed; however, amounts of energy consumed per day had dramatic effects on the proportion of the ME from each gram of feed that was used for maintenance, product, or heat increment. A model was developed that showed these effects and the feed:gain ratio over a wide range of daily energy intake.

Cloning and expression of delta-like protein 1 messenger ribonucleic acid during development of adipose and muscle tissues in chickens.

Delta-like protein 1 (DLK1) is involved in adipose and muscle development as shown by the reduction of fat mass in DLK1 transgenic mice and in muscle hypertrophy of callipyge sheep. However, no study on DLK1 has been investigated in avian species. Cloning and sequencing of a full length of chicken DLK1 (gDLK1) complementary DNA revealed that gDLK1 contains a total of 1,161 bp, encoding 386 amino acids. The similarity of gDLK1 nucleotide and protein sequences was over 50% compared with other mammalian species. In addition, chickens only express one full length of gDLK1 in various tissues at different ages without the alternative splicing variants of DLK1 found in mammalian species. This suggests that the full-length form of gDLK1 may be sufficient for normal development in the chicken. In adipose tissue, the gDLK1 gene was highly expressed in preadipocytes as compared with adipocytes (P < 0.05), whereas expression levels of adipogenic marker genes such as stearoyl-coenzyme A desaturase 1 (SCD-1) and fatty acid binding protein 4 (FABP4) were higher in mature adipocytes than in preadipocytes (P < 0.05 and P < 0.01, respectively). Expression of gDLK1 in adipose tissue tends to decrease with age. The expression of gDLK1 gene in the pectoralis major muscle was significantly higher in 13- and 17-d-old embryos (P < 0.05), decreased in 1- and 5-d-old chicks (P < 0.05), and further decreased in 11- and 33-d-old chickens (P < 0.05). This expression pattern of gDLK1 was very similar to the expression patterns of myogenin and Pax7 genes, suggesting a close association with myogenic activities. In conclusion, the developmental regulation of gDLK1 expression might play an important role in the early stages of adipose and muscle tissue development.

Daily energy intake of broiler chickens is altered by proximate nutrient content and form of the diet.

An experiment was designed to test the ability of broiler chickens to equalize daily energy intake when proximate components of the diet were changed. A factorial arrangement was used to test effects of protein, fat, and fiber content in the diet. The simplest diet contained only corn and soybean meal to provide energy and protein. Protein contents were calculated to be 16.4, 18.2, and 20.0%, with added protein from a combination of corn gluten meal, fish meal, and peanut meal. Hydrolyzed fat was added at 0, 2.5, 5.0, and 7.5% of the diets. A combination of alfalfa meal, oats, and wheat middlings was used to increase the fiber of the corn soy diet by approximately 2 and 4%. The 36 combinations were fed as mash. In addition, 8 of the diets were fed as pellets. All diets were fed for 12 d from the time broilers reached approximately 1.2 kg. A total excreta collection was used to determine ME, and carcass analysis provided fat and energy content. When fed mash, only sex had a significant effect on grams of feed eaten per day. Sex and dietary fat content affected gain per day. Sex, fat, and fiber altered the kcal of ME eaten per day. Broilers fed 5% added fat ate approximately 10% more energy per day than those fed no added fat, and broilers fed 4% added fiber ate approximately 20% less ME than those fed no added fiber. A comparison of results from mash and pellets showed that only sex and form affected gain per day, feed per day, and kilocalories of ME eaten per day. For the mash and pellets, protein, fat, fiber, and several interactions affected the ME per gram; however, the ME per gram was similar for pellets and mash. The results suggest that the diet composition and form have a significant effect on the energy intake of broiler chickens.

Metabolizable energy values determined with intact and cecectomized roosters.

Mature White Leghorn roosters that were intact or cecectomized were used to determine the ME of a diet. Seven intact and 7 cecectomized roosters were fasted for 3 d, with total excreta collection on d 2 and 3. Approximately 1 wk after completion of the trial, the same roosters were force-fed 28 g of a diet that was 85% corn and 15% soybean meal after a 1-d fast. Excreta were collected from d 2 and 3 and dried. There was no difference in the energy and nitrogen excreted by intact and cecectomized roosters when they were fasted for 3 d. The energy and nitrogen excreted by cecectomized roosters were greater than by intact roosters when they were fed the test diet. As a result, the AME, TME, AME(n), and TME(n) were all larger from intact roosters than from cecectomized roosters. Our results suggest that intact roosters should be used to determine the energy content of ingredients that will be fed to commercial flocks.

Amino acid availability and true metabolizable energy content of corn distillers dried grains with solubles in adult cecectomized roosters.

Five sources of corn distillers dried grains with solubles (DDGS), which varied in darkness of color, were collected from several processing plants in the Midwestern United States. Sources of DDGS were analyzed for their amino acid and energy contents, measured for color score, and evaluated for TMEn, apparent amino acid digestibility, and true amino acid digestibility. A precision-fed rooster assay was used, in which each DDGS sample was tube fed (25 g) to adult cecectomized roosters, and the excreta were collected for 48 h. The experiment was conducted as a randomized complete block design with 8 replicates. Seven adult roosters (averaging 75 wk of age) were used in each period, with 5 fed the DDGS sources and 2 fasted to estimate basal endogenous amino acid losses. One source (no. 5) was the darkest, 2 sources (no. 2 and 4) were light, whereas 2 other sources (no. 1 and 3) were intermediate in color as measured by a colorimeter. Total lysine content of the DDGS sources ranged from 0.48 to 0.76%, with the lowest lysine content in the darkest DDGS source. Apparent and true lysine digestibility was approximately 30 and 15 percentage units lower (P < 0.05), respectively, in the dark-colored source (no. 5) than in the other 4 sources. Average apparent and true digestibility of the essential amino acids were 10 and 8 percentage units lower (P < 0.05), respectively, in source 5 than the other 4 sources. The TMEn content of the 5 DDGS sources was also lower (P < 0.05) in the darkest DDGS (no. 5). Our results suggest that when the color score of a DDGS source, as measured by a colorimeter, reached a certain threshold (lightness between 28 and 34), amino acid availability and true metabolizable energy content may be reduced. This reduction was particularly evident for lysine, which had the lowest digestibility in the darkest DDGS source. These results suggest that dark-colored DDGS may have been overheated during drying, causing Maillard reactions to be more extensive and resulting in a lowered total lysine content, lysine digestibility, and TMEn content.

Myosin heavy chain isoform expression is not altered in the pectoralis major muscle in selenium-deficient chickens recovering from exudative diathetic myopathy.

The recovery of broiler chickens experiencing skeletal muscle myopathy caused by a selenium deficiency was compared with control broiler chickens in an age matched study by ultrastructural analysis of the pectoralis major (PM) muscle and examination of the temporal expression of the developmental fast skeletal myosin heavy chain (MyHC) isoforms. Selenium-deficient chicks showing signs of exudative diathesis (ED) were injected subcutaneously with sodium selenite in water and allowed to recover. At 0, 2, 5, 10, 20, and 30 d after selenium injection, a sample of the PM muscle was removed from selenium-deficient and control chicks for analysis. Ultrastructural analysis revealed vacuolization in the PM of selenium-deficient chicks with little or no visible damage to the sarcomere. Relative amounts of chicken ventricular, embryonic, neonatal, and adult fast skeletal MyHC isoforms were determined using chicken fast skeletal MyHC isoform specific monoclonal antibodies. The temporal expression of the developmental MyHC isoforms was similar in all chickens (P > 0.05). There was no expression of chicken ventricular MyHC observed in the PM of either group. These results indicate that chicken fast muscle recovering from exudative diathetic myopathy does not use the same pathways as chicken skeletal fast muscle regenerating from physical or toxic injury in which temporal expression of the MyHC isoforms is initially predominantly ventricular, then predominantly embryonic, neonatal, and finally predominantly adult developmental MyHC isoform.

Selenium toxicity in breeding ring-necked pheasants (Phasianus colchicus).

A flock of breeding ring-necked pheasants received feed with a high selenium content. Within 4 days of eating the toxic feed, the rate of egg production began to decrease, and bird aggression increased. Approximately 12% of the hens died within a week. Necropsy of the hens revealed colorless fluid around the heart and a friable, but otherwise normal, liver. The rapid onset of the problem and signs noted at necropsy suggested toxicosis. Based on analysis, the feed contained 9.3 ppm of selenium. Selenium toxicity was consistent with the histologic diagnosis of degenerative cardiomyopathy, vacuolar degeneration of hepatocytes, and centrilobular hepatic necrosis. After 8 days, the toxic feed was removed and replaced with fresh feed. Egg production, which had dropped to 50%, returned to normal within 10 days of feed replacement. Hatchability of eggs laid from days 8 to 14 after delivery of the toxic feed was 35%. Approximately 10% of the chicks that hatched had deformed beaks and abnormal eyes. Many of the chicks that died in the shell had deformities, bringing the total to more than 50% of all embryos that developed. The selenium content of eggs that had no embryonic development was 2.05 ppm. Hatchability of eggs laid from days 21 to 28 after the toxic feed was delivered was almost 80%, which was slightly lower than normal. The selenium content of these eggs was 0.30 ppm. These results show the rapid onset and correction of selenium toxicity and suggest that specific embryologic defects are diagnostic for selenium toxicity.

Estimating body weight and body composition of chickens by using noninvasive measurements.

The major objective of this research was to develop equations to estimate BW and body composition using measurements taken with inexpensive instruments. We used five groups of chickens that were created with different genetic stocks and feeding programs. Four of the five groups were from broiler genetic stock, and one was from sex-linked heavy layers. The goal was to sample six males from each group when the group weight was 1.20, 1.75, and 2.30 kg. Each male was weighed and measured for back length, pelvis width, circumference, breast width, keel length, and abdominal skinfold thickness. A cloth tape measure, calipers, and skinfold calipers were used for measurement. Chickens were scanned for total body electrical conductivity (TOBEC) before being euthanized and frozen. Six females were selected at weights similar to those for males and were measured in the same way. Each whole chicken was ground, and a portion of ground material of each was used to measure water, fat, ash, and energy content. Multiple linear regression was used to estimate BW from body measurements. The best single measurement was pelvis width, with an R2 = 0.67. Inclusion of three body measurements in an equation resulted in R2 = 0.78 and the following equation: BW (g) = -930.0 + 68.5 (breast, cm) + 48.5 (circumference, cm) + 62.8 (pelvis, cm). The best single measurement to estimate body fat was abdominal skinfold thickness, expressed as a natural logarithm. Inclusion of weight and skinfold thickness resulted in R2 = 0.63 for body fat according to the following equation: fat (%) = 24.83 + 6.75 (skinfold, ln cm) - 3.87 (wt, kg). Inclusion of the result of TOBEC and the effect of sex improved the R2 to 0.78 for body fat. Regression analysis was used to develop additional equations, based on fat, to estimate water and energy contents of the body. The body water content (%) = 72.1 - 0.60 (body fat, %), and body energy (kcal/g) = 1.097 + 0.080 (body fat, %). The results of the present study indicated that the composition of a chicken's body could be estimated from the models that were developed.

Indicators of nutritional value of hydrolyzed feather meal.

Processing pressure and time were evaluated for their effects on feather meal protein quality. Feathers were collected from a commercial broiler plant and hydrolyzed with saturated steam in an experimental batch hydrolyzer. A constant time series (36 min) was completed to evaluate the effect of increasing pressure (207 to 517 kPa) on nutritional value. Feather meal processed at the lowest pressure had the highest nutritional value, and vice versa. True amino acid availability determined with force-fed White Leghorn cockerels demonstrated that increasing pressure decreased true available (TA) cystine (P < 0.05) more than any other amino acid. Sulfur content and acid detergent fiber were positively correlated with TA sulfur amino acid content; bulk density, 0.2% pepsin-digestible protein, and acid detergent soluble protein were negatively correlated with TA sulfur amino acid content. Increased steam pressure also resulted in decreased, undegraded intake protein. Various combinations of time (106 to 4.5 min) and pressure (207 to 724 kPa) were used to prepare a constant density series (483 kg/m3). In this series, feather meals were similar in nutritional value. There was no indication that high hydrolysis pressure was detrimental to feather meal quality, if the appropriate time was used. These results suggest that sulfur content and bulk density can be used to monitor feather meal quality.

Induction of vitamin A deficiency in turkeys.

The objective of this study was to determine the relationship between the hepatic vitamin A (VitA) level and the pathologic changes in the oropharynx and esophagus of VitA-deficient turkeys. Study turkeys were provided with a diet sufficient (11,000 IU/kg) or deficient (2750 IU/kg) in VitA from 4 to 17 wk of age. Body weight, bacterial culture, and tissues from internal organs were collected at weekly intervals. VitA deficiency causes epithelial tissue damage in poultry. This epithelial damage was seen grossly as white plaques in the oropharynx and esophagus and histologically as squamous metaplasia of mucosal glands and keratinization of epithelium. No significant difference in body weights was seen among the groups. Moreover, no pathogenic bacteria was isolated during sampling periods. Liver VitA levels declined significantly after consumption of low VitA diet for 3 wk and were depleted after 5 wk. Squamous metaplasia due to VitA deficiency developed in the esophagus after 3 wk and in the oropharynx after 4 wk of consuming a VitA-deficient diet.

Morphologic characteristics of oviducts from hens producing eggs of different Haugh units caused by genetics and by feeding vanadium as determined with computer software-integrated digitizing technology.

Magnum and isthmus morphologic characteristics (surface epithelium height, fold height and diameter, and periodic acid-Schiff-positive area and surface epithelium cells) of stained 6-micron tissue sections were measured by light microscopy, with data acquisition using a digitizing tablet interfaced to a microscope and to a personal computer with morphometric-dimensional software. Tissues were obtained from Leghorn layers in two separate experiments in which production of eggs with low and high Haugh unit (HU) values was induced by either genetic selection or by feeding V. Eggs produced by these hens had HU differences between the high and low groups of 11 to 14 units (both experiments, P = .0001) and had a greater volume of thick albumen fraction in high-HU groups (both experiments, P = .0001). The computer software-integrated digitizing system enabled rapid measurements of multiple characteristics. In the genetic lines, higher magnum fold height and magnum and isthmus surface epithelium height were detected at moderate significance (all at P < .05) in the tissues of the layers producing high-HU eggs than in tissues from the low-HU line. Other morphologic variables were not different between genetic lines. In response to feeding V, none of the morphological characteristics were affected, although magnum fold height approached difference at P < .07. Based on the observations in these two experiments, magnum fold height may be a further important factor related to egg albumen condition, in addition to surface epithelium height. It appears, however, that layers producing eggs of considerably different HU values, due in these experiments to genetics or V feeding, can have magnum or isthmus morphological characteristics that are indistinguishable, or only moderately different, as detected by integrated digitizing technology.

Evidence of multiple metabolic routes in vanadium's effects on layers. Ascorbic acid differential effects on prepeak egg production parameters following prolonged vanadium feeding.

The development of V toxicity was followed over a 28-d period in 25-wk-old Leghorn layers fed 20 mg ammonium metavanadate/kg diet (Days 1 to 14) followed by 30 mg/kg diet (Days 15 to 28). Then, over a second 28-d period, the responses to V and supplemental ascorbic acid (AA) fed at 500 or 1,000 mg/kg diet (Days 29 to 42) followed by 1,500 or 3,000 mg/kg diet (Days 43 to 56) were examined. Feed consumption, egg weight, Haugh units (HU), and BW measurements indicated that the response to V was multifactorial, but of differing intensities and time-frames for the variables. Haugh units were lowered rapidly (3 d, P < .05) in response to V feeding, but HU values decreased only slightly when dietary V was increased to 30 mg/kg. In contrast, egg production was decreased moderately by 20 mg V/kg and a considerable further reduction in egg production resulted from increasing the V to 30 mg/kg. Ascorbic acid supplementation differentially affected these responses: BW, egg production, and egg weight were improved significantly in the V-fed group receiving an AA supplement, as compared with those fed V only. Haugh unit values, however, were not improved by AA supplementation in groups receiving V. Foam functional properties, which also were changed by V feeding, were not corrected by AA feeding. The results suggest that the toxic effects of V are mediated through more than one physiological mechanism. One mechanism, which includes negative effects on BW, egg production, and egg weight, is responsive to the additional reducing equivalents provided by supplemental AA. Another mechanism, which is apparent from the effect of V on egg HU values, is not ameliorated by AA supplementation after toxicity developed.

Dietary lysine concentrations from deficient to excessive and the effects on broiler chicks.

1. An experiment was designed to test the response of broiler chicks (0-21 d) to dietary lysine concentration. Concentrations ranged from 9.9 to 14.4 g of lysine per kg diet when energy was 13.4 MJ ME/kg. 2. Estimates of the concentration of lysine needed for maximum body weights gain, food consumption and gain:food ratio were calculated using two statistical methods. An average of these estimates was 12.0 g lysine/kg diet to 21 d of age. 3. Chicks given 13.9 or 14.4 g lysine/kg diet were negatively affected by these concentrations. The decreases in average weight gain, food consumption and food efficiency were caused mainly by several chicks that developed severe leg problems and were much smaller than their pen mates. Chicks with no leg problems gained weight as rapidly as chicks receiving optimal amounts of lysine.

Lysine requirement of broiler chicks as affected by protein source and method of statistical evaluation.

1. An experiment was designed to test if the lysine requirement, expressed as g lysine/kg CP, was the same for several protein sources. 2. Groundnut meal, groundnut meal adjusted with indispensable amino acids or sesame meal supplied the dietary CP at 180 g/kg diet. Increments of lysine (1.5 g/kg diet) were added to each of these diets. 3. The gain, food intake and food efficiency responses of broiler chicks were analysed using a quadratic equation and a two-slope method. An estimate of lysine requirements was also obtained from a survey of college students. 4. The different methods produced widely different estimates of lysine requirement. 5. The average lysine requirement was estimated at 50.1 g lysine/kg CP for groundnut meal, 61.7 for adjusted groundnut meal and 54.9 for sesame meal. 6. Reasons for the effect of statistical analysis and protein source on lysine requirement are discussed.

Isoleucine requirement of broiler chicks as affected by the concentrations of leucine and valine in practical diets.

1. Chicks were fed on an isoleucine-deficient diet, with 6 added concentrations of isoleucine to determine their isoleucine requirement and an additional 6 treatments were devised to determine their isoleucine requirement when dietary leucine and valine contents were increased. 2. The diet deficient in isoleucine contained 5.6 g/kg isoleucine with leucine and valine contents of 20.1 and 10.3 g/kg, respectively. Supplementation with leucine and valine increased these to 24.7 and 12.6 g/kg, respectively. 3. The isoleucine requirement was not affected by dietary leucine and valine contents in a diet with 13.4 MJ of ME per kg. Analysis of variance and Least Significant Difference of means indicated an isoleucine requirement of 7.2 g/kg. Non-linear regression of the same data indicated an isoleucine requirement of 8.44 g/kg, based on weight gain, or 8.19 g/kg based on food efficiency. 4. Reasons for the failure to find an imbalancing effect of branched chain amino acids in practical diets are discussed.

Metabolic and economic efficiency of broiler chicks as affected by dietary protein levels.

1. An experiment was designed to determine if decreasing excess amino acids in the diets of chicks would improve metabolic efficiency, as indicated by growth rate and food efficiency. 2. Semi-purified diets were fed with crude protein contents of 180 or 230 g/kg. The sources of protein were maize gluten meal, groundnut meal, sesame meal or soya-bean meal. Crystalline amino acids were supplemented to meet all amino acid requirements. 3. The dietary crude protein content had no statistically significant effect on weight gain or food consumption. The probability that the higher protein improved gain per food was 0.06. Protein source had a significant effect on all the responses measured. 4. Decreased concentrations of excess amino acids in chick diets had no favourable effects on weight gain or gain per food. Lower protein diets were more expensive per unit of gain.