Effects of egg product on growth performance, muscle nutrients, and intestinal microflora of Procambarus clarkii
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摘要: 为评估蛋粉在克氏原螯虾 (Procambarus clarkii) 饲料中的应用效果，使用喷雾干燥蛋粉等蛋白替代0% (CON)、25% (R1)、50% (R2)、75% (R3) 和100% (R4) 鱼粉，饲喂幼虾 [初始平均体质量 (9.02±0.12) g] 6周。结果显示，各组间特定生长率、饲料系数、摄食率、成活率和肝体比均无显著性差异 (P>0.05)，R4组体质量增长率较CON组显著降低 (P<0.05)。R4组肌肉粗蛋白、必需氨基酸和总氨基酸含量显著升高 (P<0.05)，R2、R3和R4组肌肉粗脂肪和灰分含量显著升高 (P<0.05)，各组肌肉中水解氨基酸含量无显著性差异 (P>0.05)。蛋粉添加组肠道蛋白酶活性显著升高，淀粉酶活性显著降低，R3和R4组肠道脂肪酶活性显著升高 (P<0.05)。R2、R3和R4组血清碱性磷酸酶活性以及总蛋白含量显著升高 (P<0.05)。R3组肠道梭状芽孢杆菌属 (Clostridium) 和溶杆菌属 (Lysobacter) 细菌丰度显著升高，R4组链球菌属 (Streptococcus) 丰度显著升高 (P<0.05)。综上，当克氏原螯虾饲料含6%鱼粉时，蛋粉可替代75%鱼粉。Abstract: To evaluate the application effect of spray-dried egg product in the diet of Procambarus clarkii, we formulated five isonitrogenous and isoenergetic diets containing 0% (Control group), 2.0%, 4.0%, 6.0% and 8.0% spray-dried egg product, which replaced 0% (CON), 25% (R1), 50% (R2), 75% (R3) and 100% (R4) fishmeal, respectively, to feed the shrimps [Initial body mass (9.02±0.12) g] for six weeks. The results show that there were no significant differences in the specific growth rate, feed conversion ratio, feeding rate, survival rate and hepatosomatic index among all the groups (P>0.05). The weight gain rate in R4 group decreased significantly compared with the CON group (P<0.05). The contents of crude protein, essential amino acid and total amino acid in abdominal muscle in R4 group increased significantly compared with the CON group (P<0.05). The contents of crude fat and ash in muscle in R2, R3 and R4 groups were significantly higher than those in the CON group (P<0.05). There were no significant differences in the contents of 17 hydrolyzed amino acids in muscle among all the groups (P>0.05). Compared with the CON group, the intestinal protease activities increased significantly but the amylase activities decreased significantly in egg product supplementation groups, and the intestinal lipase activity increased significantly in R3 and R4 groups (P<0.05). The serum alkaline phosphatase activity and total protein content in R2, R3 and R4 groups increased significantly than those in the CON group (P<0.05). The bacterial abundances of Clostridium sensu stricto and Lysobacter in R3 group were significantly higher than those of the other groups, while the bacterial abundance of Streptococcus in R4 group was significantly higher than that of the other groups (P<0.05). In conclusion, when the diet of P. clarkii contains 6% fishmeal, the spray-dried egg product can replace 75% fishmeal.
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Key words:
- Procambarus clarkii /
- Spray-dried egg product /
- Intestinal microbiome /
- Immunity /
- Growth
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图 1 克氏原螯虾肠道微生物样品测序稀释曲线
Figure 1. Sequencing dilution curve of gut microbial samples from P. clarkii
图 3 克氏原螯虾肠道微生物在门水平 (a、b) 和属水平 (c、d) 上的群落组成
Figure 3. Community composition of gut microbes of P. clarkii at phylum level (a, b) and genus level (c, d)
图 4 克氏原螯虾肠道微生物在门水平 (a) 和属水平 (b) 上差异菌群分析
Figure 4. Differential flora analysis of gut microbes of P. clarkii at phylum level (a) and genus level (b)
表 1 蛋粉和鱼粉的营养组成成分 (干物质基础)
Table 1. Nutritional components of egg product and fishmeal (Dry matter basis)
% 营养组成 Nutritional composition 蛋粉Egg product m (氨基酸)∶m (总蛋白)Amino acid : Total protein 鱼粉Fishmeal m (氨基酸)∶m (总蛋白)Amino acid : Total protein 干物质 Dry matter 95.32 91.46 粗蛋白质 Crude protein 50.23 67.04 粗脂肪 Crude lipid 34.98 7.54 灰分 Ash 4.88 15.37 无氮浸出物 Carbohydrates 6.34 0.46 可溶性寡肽① Soluble peptides 5.25 7.34 精氨酸 Arginine 3.01 5.99 3.76 5.61 组氨酸 Histidine 1.09 2.17 1.44 2.15 异亮氨酸 Isoleucine 2.43 4.84 2.50 3.73 亮氨酸 Leucine 4.34 8.64 4.39 6.55 赖氨酸 Lysine 3.37 6.71 4.75 7.09 蛋氨酸 Methionine 1.72 3.42 1.63 2.43 胱氨酸 Cystine 1.36 2.71 0.49 0.73 苯丙氨酸 Phenylalanine 2.59 5.16 2.55 3.80 苏氨酸 Threonine 2.26 4.50 2.39 3.57 色氨酸 Tryptophan 0.50 1.00 0.65 0.97 缬氨酸 Valine 3.21 6.39 2.94 4.39 酪氨酸 Tyrosine 1.84 3.66 1.97 2.94 总必需氨基酸 ΣEAA 24.52 48.82 27.00 40.27 注：①. 可溶性寡肽指可溶于水的易被吸收的小肽。 Note: ①. Soluble oligopeptides are small peptides which are soluble in water and can be directly absorbed. 
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表 2 实验饲料基础配方 (干物质基础)
Table 2. Ingredients of experimental diets (Dry matter basis)
% 原料 Ingredient 替代鱼粉水平 Substitution levels of fishmeal 0% (CON) 25% (R1) 50% (R2) 75% (R3) 100% (R4) 鱼粉 Fishmeal 6.00 4.50 3.00 1.50 0.00 蛋粉 Spray-dried egg product 0.00 2.00 4.00 6.00 8.00 小麦蛋白粉 Wheat gluten 8.00 8.00 8.00 8.00 8.00 豆粕 Soybean meal 20.00 20.00 20.00 20.00 20.00 菜粕 Rapeseed meal 6.00 6.00 6.00 6.00 6.00 棉粕 Cotton meal 11.00 11.00 11.00 11.00 11.00 面粉 Flour 25.00 25.00 25.00 25.00 25.00 鱼油 Fish oil 2.00 2.00 2.00 2.00 2.00 大豆油 Soybean oil 2.00 1.60 1.20 0.80 0.40 大豆磷脂油 Soybean phospholipid oil 1.00 0.90 0.80 0.70 0.60 胆固醇 Cholesterol 0.50 0.40 0.30 0.20 0.10 酵母膏 Yeast extract 2.00 2.00 2.00 2.00 2.00 磷酸二氢钙 Ca(H2PO4)2 2.30 2.30 2.30 2.30 2.30 维生素预混料① Vitamin premix 1.00 1.00 1.00 1.00 1.00 无机盐预混料① Mineral premix 1.00 1.00 1.00 1.00 1.00 褐藻酸钠 Sodium alginate 2.00 2.00 2.00 2.00 2.00 氯化胆碱 Choline chloride 0.20 0.20 0.20 0.20 0.20 虾青素 Astaxanthin 0.04 0.04 0.04 0.04 0.04 壳聚糖 Chitosan 0.10 0.10 0.10 0.10 0.10 赖氨酸 Lysine 0.40 0.40 0.40 0.40 0.40 苏氨酸 Threonine 0.38 0.38 0.38 0.38 0.38 微晶纤维素 Cellulose 6.78 6.88 6.98 7.08 7.18 膨润土 Amargosite 2.30 2.30 2.30 2.30 2.30 总计 Total 100 100 100 100 100 注：①. 维生素和矿物元素预混料的配制参照本研究室杨文秀等[12]。 Note: ①. The preparation of the vitamin and mineral element premix is referred to the previous report of this laboratory[12].
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表 3 实验饲料营养组成 (干物质基础)
Table 3. Nutritional composition of experimental diets (Dry matter basis)
% 营养组成 Nutritional composition 替代鱼粉水平 Substitution level of fishmeal 0% (CON) 25% (R1) 50% (R2) 75% (R3) 100% (R4) 干物质 Dry matter 88.74 90.10 89.83 89.97 89.70 粗蛋白质 Crude protein 35.27 35.34 35.22 34.94 35.27 粗脂肪 Crude fate 5.92 5.68 6.03 6.08 6.14 灰分 Ash 8.21 8.31 8.35 8.19 8.29 总能 Gross energy/(kJ·g−1) 19.87 19.56 19.65 19.77 19.78 必需氨基酸 EAA 精氨酸 Arginine 1.38 1.48 1.49 1.43 1.43 组氨酸 Histidine 0.55 0.54 0.56 0.53 0.54 异亮氨酸 Isoleucine 0.87 0.82 0.85 0.81 0.81 亮氨酸 Leucine 0.61 0.62 0.63 0.61 0.62 赖氨酸 Lysine 2.62 2.51 2.55 2.58 2.59 蛋氨酸 Methionine 0.26 0.29 0.29 0.26 0.25 苯丙氨酸 Phenylalanine 1.19 1.20 1.25 1.28 1.19 苏氨酸 Threonine 1.77 1.75 1.79 1.77 1.74 缬氨酸 Valine 1.36 1.38 1.49 1.46 1.48 非必需氨基酸 NEAA 丙氨酸 Alanine 1.02 0.98 0.99 0.94 0.93 天冬氨酸 Aspartic acid 3.27 3.34 3.36 3.33 3.32 胱氨酸 Cysteine 0.25 0.29 0.29 0.28 0.30 甘氨酸 Glycine 1.53 1.40 1.30 1.17 1.11 谷氨酸 Glutamate 7.67 7.50 7.52 7.49 7.49 脯氨酸 Proline 3.51 2.75 2.90 2.82 2.81 丝氨酸 Serine 1.85 1.88 1.97 1.98 1.96 酪氨酸 Tyrosine 1.62 1.59 1.63 1.54 1.64 总必需氨基酸 ΣEAA 10.61 10.61 10.89 10.73 10.44 总非必需氨基酸 ΣNEAA 20.70 19.72 19.96 19.55 19.57 总氨基酸 ΣTAA 31.31 30.33 30.85 30.28 30.21
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表 4 饲料蛋粉替代鱼粉对克氏原螯虾生长性能的影响
Table 4. Effect of fishmeal replaced with spray-dried egg product on growth performance of P. clarkii
项目 Item 替代鱼粉水平 Substitution level of fishmeal 0% (CON) 25% (R1) 50% (R2) 75% (R3) 100% (R4) 初始体质量 IBW/g 9.31±0.22 9.34±0.20 9.34±0.43 9.12±0.20 9.25±0.37 终末体质量 FBW/g 23.70±0.56b 23.76±0.78b 23.40±0.99b 21.93±0.49ab 21.63±0.63a 体质量增长率 WGR/% 154.39±2.35b 154.26±6.24b 150.51±4.61b 137.35±8.59ab 137.13±8.57a 成活率 SR/% 77.78±5.56 77.79±5.55 83.33±5.56 85.18±3.20 79.63±3.20 饲料系数 FCR 1.35±0.05 1.37±0.08 1.39±0.04 1.31±0.04 1.35±0.04 摄食率 FR/% 3.36±0.14 3.39±0.26 3.46±0.16 3.23±0.29 3.30±0.04 肝体比 HSI/% 5.44±0.56 5.82±0.23 5.77±0.63 5.36±0.91 5.74±1.39 腹部含肉率 FC/% 11.78±1.76c 11.40±1.32bc 10.44±1.91abc 9.85±1.47ab 9.28±0.39a 注：同行数据中上标字母不同表示差异显著 (P<0.05)，下同。 Note: The values with different superscript letters within the same line are significantly different (P<0.05). The same below.
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表 5 饲料蛋粉替代鱼粉对克氏原螯虾腹部肌肉基本成分及水解氨基酸含量的影响
Table 5. Effect of fishmeal replaced with spray-dried egg product on abdominal muscle composition of P. clarkii
% 项目Item 替代鱼粉水平 Substitution level of fishmeal 0% (CON) 25% (R1) 50% (R2) 75% (R3) 100% (R4) 基本成分 Basic ingredient 水分 Moisture 77.67±0.82 77.38±1.44 78.21±2.55 76.97±0.81 76.14±0.62 粗蛋白质 Crude protein 18.60±0.15a 18.83±0.11a 18.71±0.12a 18.69±0.13a 19.06±0.21b 粗脂肪 Crude fat 0.20±0.006a 0.24±0.007ab 0.31±0.004c 0.40±0.006d 0.44±0.005d 灰分 Ash 1.21±0.07a 1.24±0.05ab 1.28±0.02bc 1.30±0.03c 1.32±0.06c 水解氨基酸 Hydrolyzed amino acid 必需氨基酸 EAA 精氨酸 Arginine 1.83±0.04 1.83±0.02 1.78±0.16 1.77±0.13 1.79±0.10 组氨酸 Histidine 0.38±0.04 0.38±0.02 0.35±0.05 0.33±0.05 0.38±0.05 异亮氨酸 Isoleucine 0.73±0.05 0.76±0.01 0.75±0.08 0.70±0.10 0.77±0.11 亮氨酸 Leucine 1.26±0.04 1.30±0.05 1.26±0.04 1.24±0.08 1.32±0.08 赖氨酸 Lysine 1.32±0.03 1.32±0.07 1.28±0.04 1.30±0.05 1.34±0.05 蛋氨酸 Methionine 0.28±0.02 0.29±0.02 0.30±0.03 0.32±0.03 0.32±0.03 苯丙氨酸 Phenylalanine 0.80±0.02 0.83±0.04 0.80±0.04 0.82±0.05 0.83±0.03 苏氨酸 Threonine 0.71±0.03 0.74±0.02 0.70±0.07 0.69±0.07 0.74±0.06 缬氨酸 Valine 0.77±0.04 0.79±0.03 0.77±0.05 0.75±0.06 0.79±0.06 非必需氨基酸 NEAA 丙氨酸 Alanine 0.86±0.02 0.90±0.04 0.86±0.06 0.85±0.05 0.91±0.04 天冬氨酸 Aspartic acid 1.43±0.04 1.44±0.06 1.39±0.06 1.40±0.10 1.46±0.08 半胱氨酸 Cysteine 0.10±0.006 0.11±0.002 0.11±0.008 0.11±0.003 0.10±0.004 谷氨酸 Glutamic acid 2.06±0.06 2.03±0.06 1.94±0.20 1.96±0.13 1.93±0.10 甘氨酸 Glycine 0.74±0.01 0.73±0.03 0.75±0.07 0.71±0.05 0.73±0.06 脯氨酸 Proline 0.60±0.04 0.55±0.03 0.55±0.06 0.52±0.03 0.56±0.03 丝氨酸 Serine 0.74±0.02 0.74±0.03 0.71±0.03 0.70±0.04 0.74±0.05 酪氨酸 Tyrosine 0.74±0.04 0.77±0.04 0.74±0.05 0.74±0.06 0.77±0.06 总必需氨基酸 ΣEAA 5.87±0.21a 6.03±0.23a 5.84±0.24a 5.87±0.28a 6.60±0.23b 总非必需氨基酸 ΣNEAA 9.48±0.24 9.48±0.29 9.54±0.62 9.57±0.67 9.48±0.53 总氨基酸 ΣTAA 15.35±0.44a 15.51±0.52a 15.38±0.56a 15.44±0.65a 16.08±0.46b
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表 6 饲料蛋粉替代鱼粉对克氏原螯虾肠道消化酶活性的影响
Table 6. Effect of fishmeal replaced with spray-dried egg product on intestine digestive enzymes activities of P. clarkii
项目 Item 替代鱼粉水平 Substitution level of fishmeal 0% (CON) 25% (R1) 50% (R2) 75% (R3) 100% (R4) 蛋白酶活性 Protease/(U·g−1) 4.61±0.28a 5.26±0.14bc 5.41±0.28c 5.01±0.27b 5.23±0.16bc 淀粉酶活性 Amylase/(U·mg−1) 2.36±0.14d 2.01±0.06c 1.89±0.03b 1.83±0.07ab 1.79±0.03a 脂肪酶活性 Lipase/(U·g−1) 14.77±0.96a 15.49±0.87a 16.80±0.47ab 17.81±0.87b 18.63±0.71c
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表 7 饲料蛋粉替代鱼粉对克氏原螯虾血清生化指标的影响
Table 7. Effect of fishmeal replaced with spray-dried egg product on hemolymph biochemical indexes of P. clarkii
项目 Item 替代鱼粉水平 Substitution level of fishmeal 0% (CON) 25% (R1) 50% (R2) 75% (R3) 100% (R4) 谷草转氨酶活性 GOT/(U·L−1) 17.75±1.02 18.25±1.67 17.75±1.95 20.00±1.83 18.50±1.45 谷丙转氨酶活性 GPT/(U·L−1) 24.75±1.93 23.00±1.49 26.75±1.97 24.25±2.90 24.75±1.18 碱性磷酸酶活性 ALP/(U·L−1) 19.00±1.35a 21.25±1.59a 22.00±1.29b 27.00±2.02b 28.50±1.11b 总蛋白质量浓度 TP/(g·L−1) 54.27±1.94a 60.31±2.62a 63.54±2.13b 81.07±4.49b 69.89±2.83b 白蛋白质量浓度 ALB/(g·L−1) 0.42±0.02 0.46±0.02 0.42±0.03 0.51±0.04 0.46±0.02 总胆固醇浓度 T-CHO/(mmol·L−1) 0.12±0.01 0.12±0.03 0.10±0.02 0.11±0.03 0.13±0.01 甘油三酯浓度 TG/(mmol·L−1) 0.45±0.05b 0.30±0.07a 0.30±0.03a 0.39±0.09ab 0.45±0.06b 葡萄糖浓度 GLU/(nmol·mL−1) 0.34±0.04 0.43±0.08 0.39±0.05 0.39±0.05 0.39±0.06
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