• Title/Summary/Keyword: Vitamin D$_2$

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Marine-derived Ca-Mg complex influences lipid and glucose metabolism, serum metabolites, colostrum profile, and stress hormone in sows over four-parity periods

  • Sungbo Cho;Santi Devi Upadhaya;Woo Jeong Seok;Seyoung Mun;Haeun Lee;Rudolf H. van der Veen;Kyudong Han;In Ho Kim
    • Journal of Animal Science and Technology
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    • v.65 no.6
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    • pp.1308-1322
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    • 2023
  • Minerals is required small amounts among various nutrients, but it has a significant impact on sow longevity and reproduction performance. This study was carried out to see the beneficial effects of marine-derived Ca-Mg complex on the reproductive performance of sows during four-parity periods. Seventy-two gilts ([Yorkshire × Landrace] × Duroc), with an average body weight of 181 kg, were randomly allocated to three groups; CON (basal diet), 0.3LC (CON - MgO - 0.3% limestone + 0.4% Ca-Mg complex), and 0.7LC (CON - MgO - 0.7% limestone + 0.4% Ca-Mg complex). During parity 3 and 4, the expression level of SCD gene was lower in the umbilical cord of piglets born to 0.3LC and 0.7LC sows compared with the CON sows. During parity 2, 3 and 4, SLC2A2 and FABP4 gene expressions were higher in the umbilical cord of piglets born to 0.7LC sows and the placenta of sows from 0.3LC groups, respectively. Ca-Mg complex increased (p < 0.05) Ca and Mg concentrations in sows and their piglets' serum as well as in colostrum regardless of parities. The serum vitamin D concentration was higher (p < 0.05) in their first parity, whereas serum prolactin and estrogen concentrations were higher (p < 0.05) during the fourth and third parity, respectively. The growth hormone concentrations were higher (p < 0.05) in the piglets born to sows during the first and second parity. The fat and immunoglobulin A (IgA) concentrations in colostrum were higher (p < 0.05) during the third and fourth parity, respectively. A reduction (p < 0.05) in salivary cortisol, epinephrine, and norepinephrine concentrations was observed in 0.3LC and 0.7LC sow groups compared with CON after farrowing regardless of parity, however before farrowing, a reduction in norepinephrine was observed. Before farrowing, the epinephrine and norepinephrine concentrations were higher (p < 0.05) during the first and second parity. After farrowing, the concentration of these hormones was higher during the second parity. Taken together, sows' parity and dietary Ca-Mg complex supplementation influenced serum metabolites, colostrum nutrients, stress hormones as well as the gene expressions related to lipid and glucose metabolism.

Nutrition of Calcium and Phosphorus in Poultry Diets (닭에 대한 칼슘과 인의 영양)

  • 한인규;오상집
    • Korean Journal of Poultry Science
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    • v.8 no.2
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    • pp.55-68
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    • 1981
  • Calcium and phosphorus are not only indispensable for the bone formation and body fluids equilibrium but also are major components of egg shell. It is nutritionally important, therefore, to investigate the metabolism of calcium and phosphorus and to search for optimum requirement of calcium and phosphorus and the availability of various sources of calcium an4 phosphorus by poultry. An attempt was made to review the nutrition of calcium and phosphorus in poultry diets. 1, Calcium and phosphorus have great interrelationship with vitamin D in their metabolisms. 2. Most of the plant-origin phosphorus are existing in phytic form and it leads to low availability when used in poultry rations, although calcium and phosphorus present in animal-origin or mineral supplements are highly available in general. 3. Calcium and phosphorus requirement from existing information indicated that 1.0% calcium and 0.7% phosphorus for broiler and egg-type chicks, and 3.5% calcium and 0.4% phosphorus for laying hen. 4. It has been recommended that calcium and phosphorus level should be increased when the feed intake was decreased or when the egg Production rate was higher or when the hens are old. 5. Mono-, ci-, tri-, calcium phosphate, calcium carbonate, bone meal, limestone and oyster shell u the most readily available among various sources of calcium phosphorus supplements. Soft rock phosphate, deflourinated phosphate and gypsum are somewhat inferior to the previous ones in bioavailability. 6. The effect of particle size of calcium supplements on egg shell quality and egg production rate is not yet clearly defined but recent works showed that oyster shell is more available when it was coarse and limestone is more available when it was fine in panicle. size. 7. Present data indicated that mixed feeding of oyster shell and limestone is superior to the single feeding of each on laying performance. 8. Significant interaction between phosphorus and sodium was observed, that is, excessive sodium decreased egg production in layer and body weight growth in broiler in the low phosphorus diets but increased them in the high phosphorus diets.

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Introduction and Expression of PAP gene using Agrobacterium in Scrophularia buergeriana Miquel (Agrobacterium을 이용한 PAP 유전자의 현삼으로 도입 및 형질발현)

  • Yu, Chang-Yeon;Seong, Eun-Soo;Lim, Jung-Dae;Huang, Shan-Ai;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.2
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    • pp.156-165
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    • 2001
  • Exogeneous application of pokeweed antiviral protein (PAP), a ribosomal-inacivating protein in the cell wall of Phytolacca americana (pokeweed) protects heterologous plants from viral and fungal infection. A cDNA clone of PAP introduced into Scrophularia buergeriana Miquel by thransformation with Agrobacterium tumefaciences. For plant transformation, explants were precultured on shoot induction medium without kanamycin for 2-5 day, and then they were cocultured with Agrobacterium for 10 minutes. The explants were placed on co culture medium in dark condition, $28^{\circ}C$ for 2days. After explants were washed in MS liquid medium, they were transferred into selection medium including kanamycin 50mg/L (MS salts+1mg/ l BAP+2mg/ l TDZ+0,2mg/ l NAA+MS vitamin+3% sucrose+0.8% agar, pH5.8). From PCR analysis, NPT II band was confirmed in transgenic plant genome and showed resistance against fungi in antifungal activity test. Micro assay to which protein extracted from transgenic line were added, revealed hyphae growth inhibition and no spore germination at high concentration. The characteristics of inhibited hyphae was represented transparent and thin. Expression of PAP in transgenic plants offers the possibility of developing resistance to viral and fungal infection.

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Biochemical Composition of Marine Microalgae and Their Potential Antimicrobial Activity

  • Kim Se-Kwon;Jeon You-Jin;Kim Won-Suk;Back Ho-Cheol;Park Pyo-Jam;Byun Hee-Guk;Bai Sungchul C.
    • Fisheries and Aquatic Sciences
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    • v.4 no.2
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    • pp.75-83
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    • 2001
  • This study is to investigate biochemical compositions of two species of marine microalgae, Chlorella ellipsoidea of Chlorophyta and Tetraselmis suecica of Prasinophyta, and to assess their potential antimicrobial activities. Crude protein, lipid and carbohydrate for C. ellipsoidea were $43.15\%$, $12.63\%$ and $13.09\%$, respectively, and those for T. suecica were $44.95\%$, $4.80\%$ and $24.05\%$, respectively. The major amino acids of the two micro algae were aspartic acid, glutamic acid, glycine, alanine, valine, leucine, lysine and proline, and no significant difference between the amino acid compositions of both micro algae was observed. The major sugars for both microalgae were glucose, galactose and mannose, and glucose contents showed the highest level, $58.70\%$ for C. ellipsoidea and$57.86\%$ for T. suecica. The major mineral contents of both micro algae for 100g were Ca (3,114mg in C. ellipoidea and 3,389mg in T. suecica) and followed by Na (2,881mg), K (548mg) and Mg (545mg) for C. ellipsoidea and Na (1,832 mg), Mg (1,510mg) and K (548mg) for T. suecica. In the content of ATP-related compound, hypoxanthine in C. ellipsoidea and IMP in T. suecica were absolutely dominant compounds. The highest content of fatty acid in C. ellipsoidea was 20:4, $27.15\%$ and that in T. suecica was 18:3 (w-6), $18.10\%$. In case of physiologically important polyunsaturated fatty acids like eicosapentaenoic acid (20: 5) and docosahexaenoic acid (22: 6), both microalgae possessed just trace amounts but was rich in arachidonic acid (20: 4). Vitamin content in both microalgae was significantly high in choline and inositol. In antimicrobial activity by water- and fat-soluble fraction of the micro algae, hexane extract in the fat-soluble fraction of C. elliposidea inhibited the growth of Bacillus subtilis by $96\%$ bactericidal activity and tetrachlorocarbon extract of T. suecica indicated relatively excellent antimicrobial activity $(81\%\;bactericidal\;activity)$ against Escherichia coli. Hot water extract among water-soluble fraction of both micro algae almost suppressed the growth of Staphylococcus aureus by $96\%$ bactericidal activity.

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Identification of genes involved in inbreeding depression of reproduction in Langshan chickens

  • Xue, Qian;Li, Guohui;Cao, Yuxia;Yin, Jianmei;Zhu, Yunfen;Zhang, Huiyong;Zhou, Chenghao;Shen, Haiyu;Dou, Xinhong;Su, Yijun;Wang, Kehua;Zou, Jianmin;Han, Wei
    • Animal Bioscience
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    • v.34 no.6
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    • pp.975-984
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    • 2021
  • Objective: Inbreeding depression of reproduction is a major concern in the conservation of native chicken genetic resources. Here, based on the successful development of strongly inbred (Sinb) and weakly inbred (Winb) Langshan chickens, we aimed to evaluate inbreeding effects on reproductive traits and identify candidate genes involved in inbreeding depression of reproduction in Langshan chickens. Methods: A two-sample t-test was performed to estimate the differences in phenotypic values of reproductive traits between Sinb and Winb chicken groups. Three healthy chickens with reproductive trait values around the group mean values were selected from each of the groups. Differences in ovarian and hypothalamus transcriptomes between the two groups of chickens were analyzed by RNA sequencing (RNA-Seq). Results: The Sinb chicken group showed an obvious inbreeding depression in reproduction, especially for traits of age at the first egg and egg number at 300 days (p<0.01). Furthermore, 68 and 618 differentially expressed genes (DEGs) were obtained in the hypothalamus and ovary between the two chicken groups, respectively. In the hypothalamus, DEGs were mainly enriched in the pathways related to vitamin metabolism, signal transduction and development of the reproductive system, such as the riboflavin metabolism, Wnt signaling pathway, extracellular matrix-receptor interaction and focal adhesion pathways, including stimulated by retinoic acid 6, serpin family F member 1, secreted frizzled related protein 2, Wnt family member 6, and frizzled class receptor 4 genes. In the ovary, DEGs were significantly enriched in pathways associated with basic metabolism, including amino acid metabolism, oxidative phosphorylation, and glycosaminoglycan degradation. A series of key DEGs involved in folate biosynthesis (gamma-glutamyl hydrolase, guanosine triphosphate cyclohydrolase 1), oocyte meiosis and ovarian function (cytoplasmic polyadenylation element binding protein 1, structural maintenance of chromosomes 1B, and speedy/RINGO cell cycle regulator family member A), spermatogenesis and male fertility (prostaglandin D2 synthase 21 kDa), Mov10 RISC complex RNA helicase like 1, and deuterosome assembly protein 1) were identified, and these may play important roles in inbreeding depression in reproduction. Conclusion: The results improve our understanding of the regulatory mechanisms underlying inbreeding depression in chicken reproduction and provide a theoretical basis for the conservation of species resources.

Effects of Fertilization Time and Culture Medium of Pig Oocytes Matured In Vitro by liquid Boar Sperm Stored at $4^{\circ}C$ (체외성숙된 돼지난포란을 $4^{\circ}C$ 보존 액상정액으로 체외수정시 수정시간과 배양배지의 영향)

  • Park, C. S.;Y. J. Yi;Kim, M. Y.;Y. J. Chang;Lee, S. H.;D. I. Jin
    • Korean Journal of Animal Reproduction
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    • v.27 no.3
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    • pp.215-223
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    • 2003
  • This study was to investigate the effects of fertilization time and culture medium of pig oocytes matured in-vitro by liquid boar sperm. The sperm rich fraction (30∼60 ml) was slowly cooled to room temperature (20∼23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min 800 ${\times}$ g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of the LEN diluent to provide 1.0${\times}$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$. The medium used for oocyte maturation was TCM-199 supplemented with 26.19 mM sodium bicarbonate, 0.9 mM sodium pyruvate, 10 $\mu\textrm{g}$/ml insulin, 2 $\mu\textrm{g}$/ml vitamin B$_{12}$ , 25 mM HEPES, 10 $\mu\textrm{g}$/ml bovine apotransferrin, 150 $\mu$M cysteamine, 10 IU/ml PMSG, 10 IU/ml hCG, 10 ng/ml EGF, 0.4% BSA, 75 $\mu\textrm{g}$/ml sodium penicillin G, 50 $\mu\textrm{g}$/ml streptomycin sulfate and 10% pFF. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9 h in 500 ${mu}ell$ mTBM fertilization media with 1.0${\times}$10$^{6}$ sperm/ml concentration, respectively. Thereafter, oocytes were transferred into 500 ${mu}ell$ NCSU-23, HEPES buffered NCSU-23, PZM-3 and PZM-4 culture media, respectively, for further culture of 6, 48 and 144 h. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times for 6 and 9 h than in those for 1 and 3 h. The rates of cleaved oocytes were higher in the fertilization times for 6 and 9 h (85.0 and 84.6%) than in those for 1 and 3 h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time for 6 h (33.6%) than in that for 1, 3 and 9 h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9, 27.6, 26.3 and 24.4 in the fertilization times for 6, 9, 3 and 1 h, respectively. The rate of blastocyst from the cleaved oocytes and the number of cells per blastocyst were higher in HEPES buffered NCSU-23 culture medium than in NCSU-23, PZM-3 and PZM-4 culture media. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend the coincubation time of 6 h in 500 ${mu}ell$ TBM fertilization medium with 1${\times}$10$^{6}$ sperm/ml concentration and the HEPES buffered NCSU-23 culture medium for in-vitro fertilization of pig oocytes matured in-vitro.

Effects of Dietary Quercetin on Growth Performance, Blood Biochemical Parameter, Immunoglobulin and Blood Antioxidant Activity in Broiler Chicks (사료 내 Quercetin의 첨가가 육계의 생산성, 혈액 생화학 특성, 혈액 내 면역글로불린과 혈액 내 항산화 인자에 미치는 영향)

  • Kim, Dong-Wook;Hong, Eui-Chul;Kim, Ji-Hyuk;Bang, Han-Tae;Choi, Ji-Young;Ji, Sang-Yoon;Lee, Wang-Shik;Kim, Sang-Ho
    • Korean Journal of Poultry Science
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    • v.42 no.1
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    • pp.33-40
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    • 2015
  • This study was conducted to investigate the effects of dietary quercetin on growth performance, blood biochemical parameters, immunoglobulin, and blood antioxidant activity in broiler chickens. Three hundred twenty one-day old Ross broilers were divided 8 treatments (C(-), basal diet; C(+), basal diet with antibiotics; vitamin E 20 IU; vitamin E 200 IU; quercetin 20 ppm; quercetin 200 ppm; methoxylated quercetin 20 ppm; methoxylated quercetin 200 ppm) with 4 replicates and 10 birds per replicate. Birds were reared for 35 days and their feed intake and weight gain were measured weekly. At 35d, eight birds of average weight from each replicate were selected for blood collection and analysis. Weight gain of birds in the groups fed quercetin was higher when compare to NC but there was no significant difference. In the serum, creatinine, BUN and AST in quercetin groups significantly decreased compared to those of control (NC and PC) (P<0.05). The contents of IgA and IgM were significantly lower in quercetin groups than those of NC (P<0.05). SOD like activity and MDA content tended to decrease in quercetin groups, however, there was no significant difference among treatments. In conclusion, supplemental quercetin to poultry diet could be positive aspect on performance and blood metabolites. Optimum adding levels was more than 20 ppm.

A Study on the Nutrients Intake and Physical Growth and Development of Junior High School Girls in Cheju Is. Area (제주지역(濟州地域) 여중학생(女中學生)의 영양실태(營養實態)와 성장발육(成長發育)에 관한 연구(硏究))

  • Hong, Yang-Ja
    • Journal of Nutrition and Health
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    • v.8 no.4
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    • pp.15-26
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    • 1975
  • The purpose of this paper is to investigate the nutrients intake situation and the physical growth and development of junior high school girls in cheju island, the peculiar area in Korea, and to study every possible correlation ketween daily food intake and growth rate of them. Further, it made an attempt to research nutrition problems for them in this area and assist in their growth and development through proper nutritional marragement. The survey was conducted in seven sample schods selected randomly from all three districts -this is Bukcheju-gun, Namcheju-gun and cheju city-of cheju Island. Four hundred eighteen girls, aged thirteen years ald, in second year grade of junior high schools, were provided with prepared questionnaires. For six days, July $16{\sim}21$ in 1975, the respondents filled out the questionnaires. They were asked to keep record of what they ate each meat. Their mitrients intake were analyzed in accordance with their daily food intake records during a six day period. On the other side, including physical growth index calculated by means of weight and hight of the respondents, their school records were also investigated. Critical Ratio as well as percentage calculation were included for statistical analysis. The findings and results abtained are summarized as follows: (1) Living circumstance a) Average numbers of brother and sister in sampled pupils' family were five. b) Niety percent of the respondents were brought up by mother's milk at their infants and 45 percent of them were weaned at the age of 13 to 24 months. c) Fourty-six percent of them revealed irregulity in their meal time. d) The most favorite main dish was boiled rice mixed other cereals (34%), next bread (31%) and boiled noodle (26%), and kimchi was the most papular side dish (64%). (2) Nutrition survey a) The average calorie intake of a pupil per day was 1,783 cal. and the total calorie intake was composed of carbohydrate 73%, pratein 15%, fat 12% respectively. b) Carolie and nutrients were lower than the recommended level, except vitamins A,$B_1$ and Niacin, c) Average protein intake per day was 68g, which was relatively near to the recommended level, but in fact, most of them came from grain which is lacking in essential amino acids. The ratio of animal protein intake to tatal protein intake was 36%. d) Their average animal food intake was proportionated in 14% in their total food intake and the cereals consumed in the largest quantity at the level of 63% out of whole food groups. (3) The results of statistical analysis a) The correlation between the calorie intake and the physical growth index revealed statistical significancy at the level of P<.005. b) The correlation between the protein intake and the physical growth index revealed statistical significancy at the level of .01

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Food & Nutrition Survey of Specific Areas in Seoul Kores (서울시내 계층별 아파트 주민의 영양실태조사)

  • Kim, S.H.;Chung, C.E.;Lee, H.K.;Cho, S.S.;Lee, Y.W.
    • Journal of Nutrition and Health
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    • v.7 no.2
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    • pp.53-63
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    • 1974
  • For the elucidation of the nutritional status of residences of apartments located in various areas in Seoul city, the food and nutrition survey was undertaken by the graduate students from Department of Food & Nutrition, Ewha Womans University in 1973. The socioeconomic stratification was mainly considered for the selection of sample apartments included in this study. Hangang Mansion, KIST, Munwhachon & Bukahyun apts were selected in this respect. A. Common living circumstances. Through the questionaries, author concluded nutritional concept on everyday food life as follows; 1. Higher income seems less effective in everyday food life on the nutritional view point than higher education does. 2. It needs to be urgently improved to be accustomed to use exact measuring concept of foods when they cook. 3. It needs to be improved to serve meals for individual portion at meal table, because the portion control at meal table will effect to national food policy. B. Nutrition Survey. Average daily calroie intake was the highest in Hangang Mansion and lowest in Bukahyun apt., in accordance with their economic living standard, but most of their calorie intakes were lower than the R.D.A. (3000ca1 for male, 2200 Cal for female). But the composition ratio of the total calorie intake was near to the ideal, carbohydrate 60%, protein 15%, fat 25%. Although there were differences in their economic living situations in urban area, most people ingested relatively good quality and large amounts of protein and fat. Vitamin A and Niacine intake exceeded the R.D.A., but the intake of Ca, Thiamine, Riboflavin, Ascorbic Acid were far less than that of R.D.A. C. food Intake. Amounts of total daily food intake were highest in Bukahyun apt. and lowest in Hangang Mansion. These showed adverse aspects to the total calorie intakes and their income levels. The ratio of cereals, $40%{\sim}50%$ of the total food intake, was higher than any other kinds of foods in all apartments. Among the other foods except cereals, there represented the highest level of meats in Hangang Mansion and KIST apt., while intake of Kimchies were highest in Munwhachon and Bukahyun apt. In the case of vegetables and fruits intake, the ratio of them in Hangang Mansion and KIST apt. were high, on the other hand the ratio of starchy roots appeared lowest in Hangang Mansion. For the main sources of protein, meats were ingested in Hangang Mansion and KIST apt., meats and fishery were used in Munwhachon apt., and beans were eaten highly in their meals in Bukahyun apt. These food contents and distributions showed the significant differences not only their economic classes but also regional characteristics.

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Different Sources and Levels of Copper Supplementation on Performance and Nutrient Utilization of Castrated Black Bengal (Capra hircus) Kids Diet

  • Mondal, M.K.;Biswas, P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.7
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    • pp.1067-1075
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    • 2007
  • Twenty eight 3-4 month old castrated Black Bengal kids (Capra hircus) were used to determine the effects of source and level of dietary copper (Cu) concentration on their performance and nutrient utilization. Cu was supplemented (0, 10, 20 and 30 mg/kg diet DM) as copper sulfate ($CuSO_4$, $5H_2O$) or copper proteinate (Cu-P). Kids were fed a basal diet containing maize (19.5%), soybean (17.0%), deoiled rice bran (56.5%), molasses (4.0%), di-calcium phosphate and salt (1.0% each) and mineral and vitamin mixture (0.5% each) supplements at 3.5% of body weight to meet NRC (1981) requirements for protein, energy, macro minerals and micro minerals, excluding Cu. The basal diet contained 5.7 mg Cu/kg, 122.5 mg Fe/kg, 110 mg Zn/kg, 0.26 mg Mo/kg and 0.32% S. $CuSO_4$ or Cu-P was added to the basal diet at the rate of 10, 20 and 30 mg/kg. Kids were housed in a well ventilated shed with facilities for individual feeding in aluminum plated metabolic cages. Blood samples were collected from the jugular vein on d 0, 30, 60 and 90 to determine hemoglobin (Hb), packed cell volume (PCV), total erythrocyte count (TEC), total leukocyte count (TLC) and serum enzymes (alkaline phosphatase, alanine transferase and aspertate transferase). A metabolism trial of 6 days duration was conducted after 90 days of experimental feeding. Statistical analysis revealed that source and level of Cu supplementation improved live weight gain (p<0.04) and average daily gain (p<0.01). No significant contribution of source and level of Cu to alter serum serum enzymes was evident. Goats fed Cu-P tended to have higher Hb, PCV and TEC than with $CuSO_4$ supplementation. Cu-P increased digestibility of ether extract (EE, p<0.02) and crude fiber (p<0.05) and showed an increasing trend (p<0.09) for digested crude protein (CP) and crude fiber (CF). Supplemental dose of Cu linearly improved (p<0.02) digestibilities of dry matter (DM), organic matter (OM), EE and nitrogen free extract (NFE). Though the absorption of nitrogen (N) was not affected (p>0.10) by both source and dose of Cu, N retention was affected (p<0.04) and there was a significant $Source{\times}Dose$ interaction (p<0.05). Final body weight (BW) was not influenced (p>0.10) by the source of Cu but increasing dose of Cu increased (p<0.04) the BW of kids. TDN intake (g/kg $W^{0.75}$) was higher (p<0.05) with the increased dose of Cu and there was a significant $Source{\times}Dose$ interaction. It was concluded that supplementation of Cu from different sources and varying dose level in a concentrate based diet may improve performance, nutrient utilization and plane of nutrition in castrated Black Bengal kids. The effects on performance and nutrient utilization are more pronounced with Cu-P than $CuSO_4$ supplementation. Higher dose of Cu showed better result than lower dose.