An experiment was conducted to elucidate the effect of graded levels of vitamin $D_3$ in White Leghorn (WL) layer breeders on egg production, shell quality, hatchability of eggs and juvenile performance of offspring during their late laying period (72-88 wk). White Leghorn breeder females were randomly divided into 5 groups of 50 each and were housed in individual California cages in an open-side housing system. Considering birds in five cages as a replicate, 10 such replicates were randomly allotted to each treatment. A basal diet was formulated containing all the nutrients as recommended for WL layers except vitamin $D_3$, which served as control. Another, four diets were formulated by supplementing graded levels of feed grade crystalline cholecalciferol to the basal diet that contained 300, 600, 1,200 and 2,400 ICU of vitamin $D_3$ per kg. Each diet was offered ad libitum to one of the above five treatment groups. The egg production, egg weight, daily feed consumption and the feed intake per dozen eggs or kg egg mass of the birds fed diet without any supplemental vitamin $D_3$ was comparable with those of supplemental groups. Similarly, the level of vitamin in the diet did not have any effect on any of the above parameters. However, the specific gravity of eggs laid by the birds fed the diet without supplemental vitamin $D_3$ was comparable with either 600 or 2,400 ICU supplemental groups but significantly higher when compared to the 300 and 1,200 ICU groups. The egg -shell breaking strength was significantly lowered in the 600 ICU supplemental groups as compared to the strength of other dietary groups. The Haugh unit, egg shell weight, shell thickness, tibia breaking strength, bone ash and calcium content were not influenced by vitamin $D_3$ concentration in the diet. Serum Ca concentration was influenced by vitamin $D_3$ level in the diet. The serum Ca concentration of birds fed either control or the vitamin supplemented diet up to 1200 ICU/kg diet was comparable. However, increasing the concentration of vitamin $D_3$ to 2,400 ICU/kg diet significantly enhanced the concentration of Ca in the serum, which was significantly higher compared to other dietary groups. The serum concentration of P and protein, however, was not influenced by level of vitamin $D_3$ in the diet. Neither fertility nor hatchability was influenced by vitamin $D_3$ concentration in the diet. Feeding a vitamin $D_3$ deficient diet or supplementation of vitamin to hens did not have any influence on their progeny chicks. It can be concluded that dietary supplementation of vitamin $D_3$ may not be essential for optimum production, shell quality, hatchability, and juvenile performance of WL breeders during 72 to 88 weeks of age.
An experiment was conducted to study the production performance of broiler breeder females (25 to 40 weeks of age) fed either reference diet or low non-phytate phosphorus (NPP) diet with or without microbial phytase (500 FYT/kg) supplementation. A weighed (160 g/b/d) quantity of feed from each diet was offered daily to 40 replicates of one bird each housed in California type cage having individual feeders. Each cage was considered as a replicate. A continuous 16-h light per day was provided using incandescent bulbs. Body weight, egg production, egg weight, feed per egg mass, egg specific gravity, egg breaking strength, shell thickness, tibia ash and serum Ca and protein concentrations were not affected by reducing the NPP level from 0.30 to 0.18% in the broiler breeder diet. Supplementation of phytase (500 FYT/kg) enzyme to the diet containing 0.18% NPP had no added advantage on any of the above production parameters. The serum inorganic P was increased significantly (p<0.05) by either enhancing the NPP content from 0.18 to 0.30% or supplementing phytase @500 FYT/kg to the diet containing low P which were found comparable. Retention of Ca and P was positive on all the diets. P retention decreased significantly (p<0.05) with either increase in NPP content or phytase supplementation in the diet. Neither NPP nor phytase supplementation influenced bone mineralization in terms of tibia ash and strength. The hatchability was not influenced by either increasing the NPP content or supplementing the enzyme phytase. Similarly, the P concentration in the egg yolk and day old chick, day old and 14th day body weight and leg score was not altered by increasing the level of NPP or supplementing phytase enzyme. The mortality was within the normal limits in all the three dietary groups. Thus, it can be concluded that 0.18% NPP (288 mg NPP intake/b/d) in the broiler breeder' diet is adequate in sustaining the optimum performance from 25 to 40 wks of age. Enhancing the NPP content or supplementation of phytase (500 FYT/kg diet) to diet containing 0.18% NPP had no added advantage on performance.
Current influenza vaccines are produced in embryonated chicken eggs. However, egg-based vaccines have various problems. To address these problems, recombinant protein vaccines have been developed as new vaccine candidates. Unfortunately, recombinant proteins frequently encounter aggregation and low stability during their biogenesis. It has been previously demonstrated that recombinantly expressed proteins can be greatly stabilized with high solubility by fusing stabilizing peptide (SP) derived from the C-terminal acidic tail of human synuclein (ATS). To investigate whether SP fusion proteins can induce protective immunity in mice, we produced influenza HA and SP fusion protein using a baculovirus expression system. In in vitro tests, SP-fused recombinant HA1 (SP-rHA1) was shown to be more stable than recombinant HA1 (rHA1). Mice were immunized intramuscularly with baculovirus-expressed rHA1 protein or SP-rHA1 protein ($2{\mu}g/mouse$) formulated with aluminum hydroxide. Antibody responses were determined by ELISA and hemagglutination inhibition assay. We observed that SP-rHA1 immunization elicited HA-specific antibody responses that were comparable to rHA1 immunization. These results indicate that fusion of SP to rHA1 does not negatively affect the immunogenicity of the vaccine candidate. Therefore, it is possible to apply SP fusion technology to develop stable recombinant protein vaccines with high solubility.
Identification of salmonellosis-infected commercial poultry flocks has become a pivotal component of efforts to reduce incidence of egg-associated transmission of S. enteritidis and S. typhimurium to humans. As a basic study for sanitary control of S. enteritidis and S. typhimurium, main food-borne pathogenic bacteria in eggs produced by domestic hens, commercial egg samples were tested for specific antibodies to whole cells of S. enteritidis and S. typhimurium and outer membrane protein(OMP) of S. typhimurium by ELISA to detect infection of S. enteritidis and S. typhimurium in various groups of hens. When the antibody titers of yolks from three commercial brand eggs were tested after diluting in the ratio from 1:100 to 1:1,600 with double dilution method, ELISA values of the specific antibodies could be shown as differences in dilution patterns by comparing with negative control egg. When the antibody titers of the yolks from two commercial brand eggs were tested after diluting in the ratio of 1:200 and 1:1,000, ELISA values of specific antibodies were different among same brand eggs. When the antibody titers of yolks from five eggs sampled randomly from twenty one commercial brand eggs were tested after diluting in the ratio of 1:1,000, ELISA value of the specific antibodies were shown generally high. ELISA values of 28.5, 30, and 28.5% of yolks from 21 brand eggs were shown low and similar to negative control egg in antibody titers to whole cells of S. enteritidis and S. typhimurium and OMP of S. typhimurium, respectively. The results demonstrated that ELISA test of egg yolk antibody could provide a highly sensitive indicator to detect contamination of S. typhimurium and S. enteritidis in poultry, and could be used effectively to reduce incidence of S. typhimurium and S. enteritidis infection in poultry.
Antheraea yamamai vitellin was purified from matured eggs by polyacryamide gel electrophoresis, also stage dependent appearance, immunological comparison and relative content of the protein were investigated. 1. Vitellogenin, the precursor of vitellin, was first detected in the larval hemolymph at the late spinning stage by polyacrylamide gel electrophoresis and immunoelectrophoresis. 2. The electrophoretic mobility of the vitellin was identical with that of Bombyx mori and of Bombyx mandarina. However, the specific antiserum against A. uamamai vitellin did not react with either that of Bombyx mori or Bombyx mandarina in immumo-diffusion test. 3. Relative content of A. yamamai vitellin to the total soluble egg protein was 46.0 percent and did not change till eight days after oviposition. But the content started to decline from ten days after oviposition and was negligible in the five or serventeen month old eggs.
Antigen delivery systems play critical roles in determining the quality and quantity of Ab responses in vivo. Induction of protective antibodies by B cells is essential in the development of vaccines against infectious pathogens, whereas production of IgE antibodies is prerequisite for investigation of allergic responses, or type 1 hypersensitivity reactions. Virus-like particles (VLPs) are efficient platforms for expression of proteins of interest in highly repetitive manners, which grants strong Ab responses to target antigens. Here, we report that delivery of hen egg lysozyme (HEL), a model allergen, through VLP could provoke strong HEL specific IgE Ab responses in mice. Moreover, acute allergic responses were robustly induced in the mice sensitized with VLPs that express HEL, when challenged with recombinant HEL protein. Our data show that antigen delivery in the context of VLPs could function as a platform for sensitization of mice and for subsequent examination of allergic reactions to molecules of interest.
A laboratory study was made to develop a simple and economic model method for the systematic determination of functional properties of 'Soy Protein Isolates (SPI)' prepared from defatted soybean meal. These are required to evaluate and to predict how SPI may behave in specific systems and such proteins can be used to simulate or replace conventional proteins. Data concerning the effects of pH, salt concentration, temperature, and protein concentration on the functional properties which include solubility, heat denaturation, gel forming capacity, emulsifying capacity, and foaming capacity are presented. The results are as follows: 1) The yield of SPI from defatted soybean meal increased to 83.9 % as the soybean meal was extracted with 0.02 N NaOH. 2) The suitable viscocity of a dope solution for spinning fiber was found to be 60 Poises by using syringe needle (0.3 mm) with 15 % SPI in 0.6 % NaOH. 3) Heat caused thickening and gelation in concentration of 8 % with a temperature threshold of $70^{\circ}C$. At $8{\sim}12\;%$ protein concentration, gel was formed within $10{\sim}30\;min$ at $70{\sim}100\;^{\circ}C$. It was, however, disrupted rapidly at $125\;^{\circ}C$ of overheat treatment. The gel was firm, resilient and self-supporting at protein concentration of 14 % and less susceptible to disruption of overheating. 4) The emulsifying capacity (EC) of SPI was correlated positively to the solubility of protein at ${\mu}=0$. At pH of the isoelectric point of SPI (pH 4.6), EC increased as concentration of sodium chloride increased. Using model system$(mixing\;speed:\;12,000\;r.p.m.,\;oil\;addition\;rate:\;0.9\;ml/sec,\;and\;temperature\;:\;20{\pm}1\;^{\circ}C)$, the maximum EC of SPI was found to be 47.2 ml of oil/100 mg protein, at the condition of pH 8.7 and ${\mu}=0.6$. The milk casein had greater EC than SPI at lower ionic strength while the EC of SPI was the same as milk casein at higher ionic strength. 5) The shaking test was used in determining the foam-ability of proteins. Progressively increasing SPI concentration up to 5 % indicated that the maximum protein concentration for foaming capacity was 2 %. Sucrose reduced foam expansion slightly but enhanced foam stability. The results of comparing milk casein and egg albumin were that foaming properties of SPI were the same as egg albumin, and better than milk casein, particularly in foam stability.
An experiment was conducted (28 to 44 weeks) to study the laying performance, shell quality, and nutrient retention of White Leghorn layers fed different levels of non-phytate phosphorus (NPP). Six levels of NPP (0.15, 0.18, 0.21, 0.24, 0.27 and 0.30%) at a constant calcium (Ca) level (3.5%) in maize-soya-deoiled rice bran based diets were formulated, and each experimental diet was offered ad libitum for 16 weeks to five replicates with five birds in each replicate. The body weight of WL layers fed diet containing 0.15% NPP was significantly (p<0.05) lower than those fed diet with 0.30% NPP, at 44 weeks of age. However, the hen day egg production, egg weight, daily feed intake and feed consumed per dozen eggs were not influenced by the variation in the NPP levels in the diet. The bone ash content was significantly (p<0.05) higher in the birds fed 0.30% NPP as compared with those fed diets up to 0.24% NPP. Bone ash content was intermediate in the birds fed diet containing 0.27% NPP. The tibia strength followed the same trend as that of bone ash. Dietary NPP content had no influence on serum Ca and protein concentration and activity of alkaline phosphatase. However, serum inorganic P concentration increased linearly with NPP content in the diet. The concentration of P was significantly (p<0.05) higher in the birds fed 0.27% NPP or higher as compared with those fed 0.15% NPP. Levels of dietary NPP had no influence on egg quality parameters like shell wt, shell thickness, shell strength and specific gravity. The retention of nutrients such as DM, N and Ca were comparable among the WL layers fed different levels of NPP. However, the retention of P decreased linearly with increase in the level of NPP in the diet. The retention of P in the birds fed diets up to 0.24% NPP in the diet was comparable, however further increasing the content of NPP (either 0.27% or 0.30%) reduced the retention of P. Based on the results of the present study, 0.15% NPP (180 mg/b/d) in the diets of WL layers is adequate for optimum production performance during 28 to 44 weeks of age, however, WL layers require 0.27% NPP (324 mg /b/d) in the diet for optimum production with better bone mineralization.
Metabolizable energy (ME) required for basal metabolism, activity and growth was considered as the criterion for targeting specific increases in body weight (100 g/week) of broiler chicks during the grower phase (5-20 weeks) and its impact was evaluated on breeder performance. Broiler female chicks (460) from a synthetic dam line were randomly distributed to 4 test groups with 23 replicates of 5 birds each and housed in cages. The first group (ME-100) was offered a calculated amount of ME by providing a measured quantity of grower diet (160 g protein and 2,600 kcal ME/kg) which increased with age and weight gain (133-294 kcal/bird/day). The other three groups were offered 10 or 20% less ME (ME-90 and ME-80, respectively) and 10% excess ME (ME-110) over the control group (ME-100). From 21 weeks of age, a single breeder diet (170 g protein and 2,600 kcal ME/kg) was uniformly fed to all groups and the impact of grower ME restriction on breeder performance evaluated up to 58 weeks. The targeted body weight gain of 1,600 g in a 16-week period was achieved by pullets of the ME-100 group almost one week earlier by gaining 8.7 g more weight per week. However, pullets in the ME-90 group gained 1,571 g during the same period, which was closer to the targeted weight. At 20 weeks of age, the conversion efficiency of feed (5.21-5.37), ME (13.9-14.1 kcal/g weight gain) and protein (0.847-0.871 g/g weight gain), eviscerated meat yield, giblet and tibia weights were not influenced by ME restriction, but the weights of abdominal fat and liver were higher with increased ME intake. Reduction of ME by 10% in the grower period significantly delayed sexual maturity (169.3 d), but increased egg production (152.5 /bird) with better persistency. Improved conversion efficiency of feed, ME and protein per g egg content were also observed in this group up to 56 weeks. The fertility and hatchability at 58 weeks of age were higher in the ME-90 group compared to the control and 10% excess ME feeding. In conclusion, the present study revealed the possibility of achieving targeted weight gain in broiler growers by feeding measured quantities of ME during the rearing period with consequential benefits in breeder performance.
Many immunostimulating substances have been developed to improve immunity of domestic animals, although their exact mode of action and effects are not clearly defined, and they are now widely used in feed industry. Bacterial lipopolysaccharides, called endotoxin, in particular may have a profound effect not only on the immune system but also on macrophages of the reticuloendothelial system. Glucans from a variety of yeast cell wall have been shown to stimulate both specific and non-specific immune responses and to increase growth performance in pigs. Recently, there has been great interest in the role of complex carbohydrates in disease prevention and treatment. Mannanoligosaccharide is a glucomannoprotein complex derived from the cell wall of yeast. Generally, it was also known that the deficiencies of some major vitamins (vitamin A, E and C) and minerals (chromium and selenium) lead to impaired immune system and, as a result, immune function is depressed and recovery delayed. On the other hand, many researchers suggested that one possible reason for the superior performance observed in pigs fed plasma protein may be because of the presence of biologically active plasma proteins (e.g., immunoglobulins) which are known to contribute to the health of the starter pig. And, immunoglobulins present in plasma protein have been implicated as contributing to the overall immunocompetence of the newborn pig. Other immunostimulants, lactoferrin and lysozyme, mainly found in milk and egg white, have been known as having bacteriocidal and bacteriolytic effect. When considering practical use of immunostimulants, the concept of using immunostimulants is new to many people and, in most cases, it is poorly understood how and why such compounds act, and how they should be used in practice. Therefore, in order to clarify the reason for discrepancies in results, special attention should be paid to the dose/response relationship of immunostimulants and the duration of the effect.
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