Effects of lysine, methionine, threonine and tryptophan on growth performance and serum antibody titers to Gumboro disease of color - Feathered chickens

Nong Lam University, Ho Chi Minh City 29 Effects of lysine, methionine, threonine and tryptophan on growth performance and serum antibody titers to Gumboro disease of color-feathered chickens Mai C. Duong1∗, Dong D. Duong2, & Huong T. N. Dang1 1Department of Veterinary Biosciences, Nong Lam University, Ho Chi Minh City, Vietnam 2Department of Animal Nutrition, Nong Lam University, Ho Chi Minh City, Vietnam ARTICLE INFO Research paper Received: April 07, 2018 Revised: May 20, 2018 Accepted: June 10, 2018 Keywords Antibody titer Broiler chickens Lysine Methionine Threonine ∗Corresponding author Duong Chi Mai Email: mai.duongchi@hcmuaf.edu.vn ABSTRACT The experiment was conducted to determine the effects of four amino acids (lysine, methionine, threonine and tryptophan) sup- plementation on growth performances and serum antibody titers to Gumboro disease (IBD, infectious bursal disease) in broiler chickens. Chicks were randomly assigned to 5 dietary groups (5 chicks/group as 12 replicates of 5 chicks) in a complete ran- domised experimental design. Group I served as control group, was fed a diet without any supplementation. Groups II and III were supplemented 10% of four amino acids (lysine, methionine, threonine and tryptophan) of the recommended requirements for 45 and 42 days, respectively. Meanwhile, Groups IV and V were supplemented 20% of four amino acids (lysine, methionine, threonine and tryptophan) of the recommended requirements for 45 and 42 days, respectively. The supplementation was started at 3 days of age in chicks. The chickens were vaccinated against Gumboro disease at day 12 and day 19. The antibody titer of the chickens in each group was assayed using IBD anti-body ELISA. Besides, the live body weight, average feed intake and feed con- version (FCR) were determined at 0, 21, 42 and 84 days of age. The results showed no significant different between groups in performance parameters such as body weight and FCR. How- ever, the present work showed highest dose of lysine, methion- ine, threonine and tryptophan supplementation for 42 days may be improved feed intake and immune response of chicken against Gumboro disease vaccination. Cited as: Duong, M. C., Duong, D. D., & Dang, H. T. N. (2018). Effects of lysine, methionine, threonine and tryptophan on growth performance and serum antibody titers to Gumboro disease of color-feathered chickens. The Journal of Agriculture and Development 17(3), 29-34. 1. Introduction In order to be effective in chicken production, preventive medicine, genetic selection and im- proved nutrition and management should be con- cerned carefully. Nutritional supplements (carbo- hydrate, protein and fats) should be provided to ensure the growth, repair of damaged tissues as well as daily maintenance. However, deficiency or excess of dietary protein or amino acids alters im- mune responses (Payne et al., 1990). Infectious bursal disease (or Gumboro disease) is one of the economically most important diseases that af- fects commercially produced chickens worldwide (Eterradossi & Saif, 2008). Chickens infected with IBDV between 3 and 6 weeks of age mostly show clinical signs and mortality accompanied with bursal atrophy. According to Muller et al. (2003), strain and the amount of the virus, age and the breed of chickens, the route of inoculation, the presence or absence of neutralizing antibodies, intercurrent primary and secondary pathogens www.jad.hcmuaf.edu.vn The Journal of Agriculture and Development 17(3) 30 Nong Lam University, Ho Chi Minh City and environmental and management factors af- fect the level of serious clinical signs in chicken. In chicken, infected with IBDV can cause immuno- suppression, which makes the birds vulnerable to a variety of secondary infections chickens also develop a poor immune response to vaccination against other pathogens (Mazariegos et al., 1990). Therefore, strict hygiene management and vac- cination programmes have been used to prevent IBD. For optimal growth and immune response, lysine and methionine are required for protein synthesis by mammals and avian species (Rubin et al., 2007). Tryptophan can be considered as a third limiting amino acid for poultry, followed by methionine and lysine (Peganova et al., 2003). According to Kidd & Hackenhaar (2006), tryp- tophan deficiency not only affects carcass quality but it also impairs the synthesis of important neu- rotransmitters such as serotonin and melatonin. Besides, threonine is a major component of in- testinal mucin and plasma gamma-globulin in an- imals (Kim et al., 1999). Thus, the aims of this study was to evaluate the effects lysine, methio- nine, threonine and tryptophan requirements on growth performances and serum antibody titer to Gumboro disease in broiler chickens. 2. Materials and Methods A total of 300 day-old chicks (DOC) of color feather breed named “Huynh De” were obtained from Binh Minh breeder farm to use in all exper- iments. The chicks were weighed on arrival, and randomly allocated into 5 equal groups with 12 replicates each (5 chicks per replicate). Feed and water were provided ad libitum. The basal diet (Table 1) was formulated to contain all essential amino acids at recommended levels (NRC, 1984), modified by Duong Duy Dong (unpublished ma- terials). In addition, chemical composition of the basal diet used in chicken feeding was shown in Table 2. Group I served as control group, was fed a basal diet without any supplementation. Groups II and III were supplemented 10% of four amino acids (lysine, methionine, threonine and tryptophan) of the recommended require- ments for 45 and 42 days, respectively. Mean- while, Groups IV and V were supplemented 20% of four amino acids of the recommended require- ments for 45 and 42 days, respectively. The sup- plementation was started at 3 days of age in chicks. The chicks were vaccinated against New- castle disease on 4, 20 and 42 days of age; and Gumboro diseases on 12 and 19 days of age. Re- sponse variables measured during the experiment included body weight, body weight gain; feed in- take, feed conversion ratio on day 0, 21 42 and 84 days of age. On 11, 18, 27, 34, 41, 49, 56, 63, 70, 77 and 84 days of age, five birds from each group were chosen at random and blood samples were collected from the brachial vein. Serum was separated by centrifugation (3000 g, 15 min) and antibody titre against IBD were performed using commercially available ELISA kits (IDEXX, Labs Inc., Westbrook, Maine, USA) according to man- ufacturer’s instructions. The data obtained were analyzed by Tukey’s test and one-way analysis of variance (ANOVA) using Minitab 16.0.A P value < 0.05 was considered statiscally significant. 3. Results and Discussion 3.1. Growth performance As shown in Table 3, the body weight and aver- age daily gain of Group III was higher than those at 84 days of age. However, no significant differ- ence about the body weight and average weight gain was found among treatment Groups. The body weights of this study reached the standards of Binh Minh company in which chickens at 100 days of age were gained from 1.7 to 1.9 kg (Vu, 2015). The highest food consumption in Group I (control group) and the lowest of this found in Group IV with the increment level of 20% for 45 days of 4-amino acid mixture supplementation were also found. Compared with Control Group, Groups I and II, it was observed that dietary treatment Groups IV and V had significant effects on the feed intake (P < 0.001) and feed conver- sion ratio (P < 0.05). Bouyeh (2012) also con- firmed that the increment levels of 10%, 20% and 30% lysine and methionine would increase body weight, cardiac and liver weight as well as de- crease feed intake of chicken. On the contrary, the increment level of 40% of these two amino acids would decrease body weight of chicken. Rogers & Pesti (1990) also reported that tryptophan- deficient diets caused a reduction in weight gain; but, the excess tryptophan in the diet caused a numerical decrease in bird weight gain (Koelke- beck et al., 1991). Therefore, the standard and the amount of amino acid consumed by the birds have a great influence on weight gain and feed intake (Teeter et al., 1993). The Journal of Agriculture and Development 17(3) www.jad.hcmuaf.edu.vn Nong Lam University, Ho Chi Minh City 31 Table 1. Ingredient composition of the experiment diets (basal diet) Ingredients (%) 0 - 21 days of age 22 – 42 days of age 43 – 84 days of age Corn 63.219 63.513 62.930 Rice bran I 5.791 2.387 28.796 Soybean meal 46 23.175 26.129 2.500 Fish oil 2.460 2.500 0.353 L-Lysin-HCl 98% 0.212 0.135 0.790 DL-Methionin 0.210 0.249 0.249 L-Threonin 98.5 0.100 0.210 0.117 L-Tryptophan 98 0.029 0,097 0.035 NaHCO3 0.817 0.820 0.805 Choline chloride 60 0.120 0.120 0.120 Antioxidants 0.020 0.020 0.020 Limestone powder 1.186 1.170 1.157 Premix BA112 0.250 0.250 0.250 Poison absorption 0.100 0.100 0.100 Herb extracts 0.015 0.015 0.015 Precursor creatine 0.060 0.060 0.060 DCP 18 2.086 2.077 2.065 Organic acids 0.100 0.100 0.100 Probiotics 0.050 0.050 0.050 Table 2. Chemical composition of the basal diet used in chicken feeding Composition Unit 0-21 days of age 22 – 42 days of age 43 – 84 days of age Dry matter % 87.699 87.628 87.581 Metabolisable energy Kcal/kg 2900.000 2900.000 2900.000 Crude protein % 19.000 18.000 17.000 Crude lipid % 5.614 5.651 5.811 Linoleic acid % 1.367 1.259 1.179 Gross fiber % 2.911 2.975 3.086 Total mineral % 5.805 5.899 6.052 Calcium % 0.950 0.950 0.950 Total phosphorus % 0.703 0.719 0.743 Available phosphorus % 0.400 0.400 0.400 Sodium % 0.226 0.230 0.230 Chlorides % 0.140 0.134 0.128 dEB meq 240.000 240.000 240.000 Total lysine % 1.174 1.090 0.996 Total methionine % 0.538 0.489 0.477 Total Met. + Cys. % 0.857 0.797 0.773 Total threonine % 0.813 0.758 0.724 Total tryptophan % 0.235 0.189 0.205 Digestible lysine % 1.080 1.000 0.910 Digestible methionine % 0.512 0.464 0.452 Digestible Met.+ Cys. % 0.778 0.720 0.697 Digestible threonine % 0.686 0.635 0.604 Digestible tryptophan % 0.210 0.166 0.182 Digestible leucine % 1.480 1.416 1.340 Digestible isoleucine % 0.707 0.665 0.619 Digestible valine % 0.777 0.738 0.695 www.jad.hcmuaf.edu.vn The Journal of Agriculture and Development 17(3) 32 Nong Lam University, Ho Chi Minh City Table 3. Body weight, average daily gain, feed intake and feed conversion ratio Group 0 day of age 21 days of age 42 days of age 84 days of age Body weight (g) I 35.25 ± 0.96 221.33 ± 15.10 602.50ab ± 35.20 1489.17 ± 96.90 II 35.75 ± 1.21 233.67 ± 12.35 617.50ab ± 35.50 1502.50 ± 99.30 III 35.00 ± 1.04 218.17 ± 19.99 621.67a ± 49.70 1520.83 ± 122.10 IV 35.66 ± 1.23 224.00 ± 15.63 575.00b ± 34.25 1510.00 ± 112.20 V 35.83 ± 0.83 228.00 ± 16.88 570.83b ± 56.00 1516.67 ± 91.10 P 0.263 0.172 0.011 0.952 Group 0 - 21days of age 22 – 42 days of age 43 – 84 days of age 0 – 84 days of age Average daily gain (g) I 8.45 ± 0.68 18.15ab ± 1.46 21.11 ± 2.27 17.10 ± 1.14 II 8.99 ± 0.53 18.27ab ± 1.60 21.07 ± 2.01 17.25 ± 1.16 III 8.33 ± 0.88 19.21a ± 2.35 21.40 ± 3.02 17.47 ± 1.43 IV 8.56 ± 0.68 16.71b ± 1.19 22.26 ± 2.57 17.34 ± 1.31 V 8.73 ± 0.76 16.32b ± 2.07 22.52 ± 2.13 17.42 ± 1.07 P 0.195 0.001 0.459 0.953 Feed intake (g/day) I 19.03b ± 2.42 41.72a ± 2.64 69.20 ± 0.37 49.43a ± 0.98 II 21.67a ± 1.72 39.92b ± 3.06 67.84 ± 2.55 48.99b ± 1.88 III 18.01b ± 1.52 39.07ab ± 3.37 68.87 ± 1.71 48.34cb ± 1.64 IV 18.97b ± 2.43 34.64bc ± 3.63 66.62 ± 1.54 46.38cb ± 1.34 V 19.08b ± 1.93 35.60c ± 4.38 68.07 ± 5.23 46.68c ± 1.49 P 0.001 0.000 0.199 0.000 FCR (kg feed/kg weight gain) I 2.25 ± 0.27 2.30 ± 0.17 3.31 ± 0.36 2.90 ± 0.21 II 2.41 ± 0.21 2.19 ± 0.19 3.24 ± 0.29 2.85 ± 0.18 III 2.18 ± 0.28 2.07 ± 0.36 3.28 ± 0.52 2.78 ± 0.26 IV 2.23 ± 0.34 2.08 ± 0.29 3.02 ± 0.33 2.69 ± 0.19 V 2.19 ± 0.18 2.20 ± 0.29 3.04 ± 0.29 2.67 ± 0.17 P 0.216 0.207 0.217 0.049 a-cMean values for control and amino-acid-supplement groups within a column not sharing a common superscript letter were significantly different at P < 0.05. 3.2. Serum antiboy titres against Gumboro disease in broiler chickens According to IDEXX laboratories (2010), the antibody titres against Gumboro disease of around 1000-4000 would be sufficient to protect chickens from this disease. As shown in Table 4, the antibody titre against Gumboro disease af- ter the first vaccination was lower than the min- imum protective of 1000 in treatment Groups, except for the high antibody titre against Gum- boro disease was also found in 18-day-old chicken of Group V (1099 ± 1144). The lower antibody titres following the primary vaccination could be due to the young age of chicken when the immune function of young animal is not well developed (Rubin et al., 2007). The higher antibody titre in Group IV was significantly different than those of the remaining Groups at 63 days of age. In ad- dition, the antibody titre in Group IV was still higher than those of the other Groups at the end of this experiment; although, no significant dif- ference was found. This observation was consent with the study conducted by Lidiya et al. (2015), the higher dose (140% of the recommended dose) of lysine and methionine improved immune re- sponse of chicken against infectious bursal dis- ease vaccination. Furthermore, the increasing to- tal methionine levels from 0.35 to 1.2% in the diet for chickens will enhance the aspects of the im- mune responses including T-cell proliferation in response to mitogen stimulation, plasma levels of immunoglobulin G; leucocyte migration and an- tibody titre (Swain & Johri, 2000). On the con- The Journal of Agriculture and Development 17(3) www.jad.hcmuaf.edu.vn Nong Lam University, Ho Chi Minh City 33 Table 4. Serum antiboy titres against Gumboro disease in broiler chickens Days of age Item Group I Group II Group III Group IV Group V P 11 (X ± SD) 1277 ± 869 939 ± 310 563 ± 470 713 ± 365 854 ± 523 0.122 CV (%) 68.02 33.05 83.38 51.19 61.28 Min 332 632 91 215 144 Max 2688 1543 1505 1334 1750 18 (X ± SD) 400ab ± 398 231.8b ± 198.6 212.9b ± 229.1 228.6b ± 267.1 1099a ± 1114 0.013 CV (%) 99.56 85.71 107.61 116.84 101.35 Min 0 0.0 0.0 0.0 260 Max 1095 486.0 559.0 828.0 3200 27 (X ± SD) 1900 ± 969 1764 ± 747 1726 ± 1171 2222 ± 1486 2181 ± 1985 0.915 CV (%) 51.00 42.35 67.83 66.87 91.00 Min 574 626 130 639 0 Max 3134 2940 3339 4869 4946 34 (X ± SD) 4019 ± 1354 3165 ± 1121 2889 ± 357 3177 ± 1621 4190 ± 1289 0.155 CV (%) 33.69 35.40 12.36 51.02 30.77 Min 2466 1416 2489 1341 2003 Max 6323 5106 3569 5290 5919 41 (X ± SD) 5163a ± 2770 3559ab ± 1463 2250b ± 1529 3718ab ± 919 2362b ± 1271 0.110 CV (%) 53.65 41.12 67.93 24.71 53.79 Min 1730 2030 144 2161 358 Max 9752 5459 4901 4994 4248 49 (X ± SD) 4470 ± 2283 4874 ± 1733 3184 ± 1440 3694 ± 830 4611 ± 1201 0.198 CV (%) 51.06 35.56 45.23 22.46 26.05 Min 2191 1760 307 2161 2891 Max 9003 6952 4625 4917 6565 56 (X ± SD) 5146 ± 1187 3951 ± 1970 4240 ± 1721 3992 ± 1182 4567 ± 1698 0.576 CV (%) 23.06 49.85 40.59 29.62 37.19 Min 2883 1083 1604 2298 2815 Max 6119 6494 6439 5941 7780 63 (X ± SD) 4328ab ± 1679 3008ab ± 1565 2819b ± 2021 5542a ± 2281 3501ab ± 1526 0.033 CV (%) 38.80 52.02 71.71 41.17 43.58 Min 1899 435 59 3131 1279 Max 7347 5534 5413 8607 6073 70 (X ± SD) 3494 ± 1770 3598 ± 1152 3134 ± 1639 5298 ± 2741 3547 ± 1599 0.178 CV (%) 50.65 32.02 52.31 51.74 45.09 Min 1706 2124 1418 3278 231 Max 6775 5229 6736 11526 5472 77 (X ± SD) 3030 ± 1742 3111 ± 1445 2753 ± 2473 4982 ± 2719 3719 ± 1008 0.192 CV (%) 57.50 46.43 89.86 54.59 27.11 Min 0 971 324 1655 1529 Max 5336 4715 7515 10121 4702 84 (X ± SD) 2983 ± 1482 2950 ± 1487 3256 ± 2399 3864 ± 2469 3191 ± 1416 0.878 CV (%) 49.69 50.42 73.69 63.89 44.38 Min 1464 693 379 1548 242 Max 6404 5652 7270 8802 4869 a-cMean values for control and amino-acid-supplement groups within a row not sharing a common superscript letter were significantly different at P < 0.05. trary, the high supplemental levels of methionine or cysteine (1.45% in the diet) were detrimental to the growth and immune responses of chickens (Tsiagbe et al., 1987), probably due to the excess production of highly toxic substances (e.g. homo- cysteine and sulphuric acid) (Wu & Meininger, 2002). 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