凡纳滨对虾对蛋白质饲料原料消化率的研究及饲料配方实践
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摘要
本文以凡纳滨对虾(Litopeaneus vannamei)为试验动物,分别采用离体消化法和体内消化法研究其对不同蛋白原料的消化率。根据蛋白原料消化率的测定结果配制鱼粉添加比例不同但可消化粗蛋白和可消化氨基酸相同的5种饲料,并进行饲养试验,通过测定生长性能、饲料表观消化率、消化酶活性和血液生化指标,评价这5种饲料的饲喂效果,探讨基于可消化蛋白质和可消化氨基酸设计饲料配方的可行性。此外,试验还探讨了花生粕替代鱼粉对凡纳滨对虾的饲喂效果。
试验一:采用离体消化法研究了凡纳滨对虾的胃、中肠腺及肠道粗酶液对鱼粉、豆粕、菜粕和花生粕的离体消化率和酶解动力学。结果表明:凡纳滨对虾不同消化部位对干物质消化率的大小顺序为:胃>中肠腺>肠道,且鱼粉在胃部的消化率显著高于中肠腺和肠道(P<0.05),花生粕在肠道的消化率显著低于胃和中肠腺(P<0.05);对蛋白质消化率的大小顺序为:中肠腺>胃>肠道,且菜粕和花生粕在肠道的消化率显著低于胃和中肠腺(P<0.05),鱼粉在肠道和胃的消化率显著低于中肠腺(P<0.05);粗酶液对4种原料干物质的消化率高低依次为:豆粕50.78%、菜粕42.02%、花生粕39.67%、鱼粉36.50%;对粗蛋白的消化率高低依次为:花生粕60.40%、豆粕56.45%、鱼粉46.28%、菜粕43.28%。在0~4 h内,粗酶液酶解4种蛋白原料时氨基酸的生成量呈增加趋势;不同消化部位酶解产生的氨基酸总量不同,高低顺序依次为中肠腺(96.72 mg)>胃(31.28 mg)>肠道(27.58 mg);不同蛋白原料酶解时的氨基酸生成速率大小顺序为花生粕(3.9154 mg/h)>鱼粉(3.4774 mg/h)>豆粕(2.8316 mg/h)>菜粕(2.7404 mg/h)。
试验二:选取12种常见蛋白质饲料原料,按照70%基础饲料+30%试验原料(血粉按85%+15%)的原则配制饲料,以三氧化二铬(Cr2O3)作为外源指示剂,测定了凡纳滨对虾(4.45±0.21 g)对蛋白原料干物质、粗蛋白、粗脂肪、灰分、能量和氨基酸的表观消化率。结果表明:各种蛋白原料干物质表观消化率在48.61%~86.98%之间;粗蛋白表观消化率在55.74%~92.35%之间;粗脂肪的表观消化率在-0.23%~92.64%之间;灰分表观消化率在60.83%~96.30%之间;总能表观消化率在51.11%~97.27%之间。鱼粉、豆粕和花生粕的总氨基酸表观消化率分别为90.56%、92.67%和90.61%,显著高于其他蛋白原料(P<0.05);玉米蛋白粉的总氨基酸表观消化率最低,仅为59.15%,显著低于其他蛋白原料(P<0.05)。12种蛋白原料总氨基酸表观消化率高低顺序为:豆粕>花生粕>鱼粉>鸡肉粉>虾头粉>饲料酵母>菜粕>肉骨粉>乌贼内脏粉>血粉>棉粕>玉米蛋白粉。
试验三:以可消化粗蛋白和氨基酸为基础,设计可消化蛋白水平和可消化赖氨酸、蛋氨酸和精氨酸水平相同、但鱼粉添加比例不同的5种饲料配方(分别为30%、25%、20%、15%和10%),配制试验饲料,并比较这些饲料对凡纳滨对虾的饲喂效果。结果表明:随着饲料中鱼粉比例下降,饲料系数和摄食率逐渐升高,蛋白质效率、干物质、粗蛋白和粗脂肪表观消化率逐渐降低。饲料中鱼粉比例为10%时,对虾增重率、存活率、特定生长率和胃蛋白酶活性显著低于其他试验组(P<0.05),肝体指数、血清尿素氮含量和溶菌酶活性显著升高(P<0.05)。利用可消化蛋白及主要限制性氨基酸配制饲料,可使饲料中鱼粉添加比例降低至15%而不影响凡纳滨对虾的生长性能、饲料利用及营养状况。
试验四:以花生粕作为鱼粉蛋白替代原料,配制6种等氮等脂的试验饲料,其中花生粕的添加比例分别为0、7、14、21、28和35%,分别替代0、15、30、45、60和75%的鱼粉蛋白,同时以一种市售凡纳滨对虾饲料做参考饲料(RD)。饲料中添加Cr2O3作为外源指示剂用于消化率测定。结果表明:花生粕在饲料中比例增加到21%以上时,增重率显著低于对照组(P<0.05);花生粕比例增加到14%以上时,饲料效率、饲料蛋白质效率和存活率显著低于对照组(P<0.05);花生粕比例增加至35%时对虾摄食率显著低于对照组(P<0.05)。随花生粕添加比例增加,虾体水分含量呈升高趋势,粗蛋白和灰分含量则呈下降趋势,但粗脂肪含量变化不明显。饲料干物质、粗蛋白、粗脂肪和灰分的消化率随花生粕添加比例增加而降低,替代比例越高,消化率下降程度则越大。对虾胃蛋白酶和肝胰蛋白酶活性,血清中的超氧化物歧化酶、过氧化物酶、酸性磷酸酶和碱性磷酸酶活性变化趋势与消化率的变化趋势一致,铜蓝蛋白含量变化趋势则相反。花生粕不同替代比例对对虾血清溶菌酶活性没有显著影响(P>0.05)。
In this paper, the nutrient digestibility of different protein feed ingredients were evaluated using in vitro and in vivo methods for Pacific white shrimp, Litopenaeus vannamei. Based on digestibility of these protein feed ingredients, five practical diets were formulated to contain the same level of digestible crude protein and digestible amino acids for the shrimp, and their effect on growth performance, body compositions, feed utilization, digestive enzyme activity and biochemical parameters were assessed. In addition, the effect of replacement fish meal by peanut meal in practical diets on growth performance for the shrimp was also investigated. The results were as follows:
Experiment 1: The digestibility of four protein feed ingredients [fish meal (FM), soybean meal (SM), rapeseed meal (RM) and peanut meal (PM)] were determined using the in vitro method and kinetics of hydrolysis of digestive enzymes in stomach, hepatopancreas and intestine of Pacific white shrimp were studied. The results showed that the order of digestion of different digestive organs for dry matter was stomach > hepatopancreas > intestine, and stomach had significantly higher dry matter digestibility of FM than hepatopancreas and intestine (P<0.05), but intestine got significantly lower dry matter digestibility of PM than stomach and hepatopancreas (P<0.05). The order of digestion of different digestive organs for crude protein was hepatopancreas > stomach > intestine, and the digestibility of PM and RM in intestin were significantly lower than in stomach and hepatopancreas (P<0.05), but the digestibility of FM in hepatopancreas was significantly higher than in stomach and intestine (P<0.05). The order of proteins for digestibility of dry matter was SM (50.78%) > RM (42.02%) > PM (39.67%) > FM (36.50%). The order proteins for digestibility of crude protein was PM (60.40%) > SM (56.45%) > FM (46.28%) > RM (43.28%). The amino acids production of enzymolysis reaction was hepatopancreas (96.72 mg) > stomach (31.28 mg) > intestine (27.58 mg); the order of enzymolysis rate for amino acid was PM (3.9154 mg/h) > FM (3.4774 mg/h) > SM (2.8316 mg/h) > RM (2.7404 mg/h).
Experiment 2: A feeding trial was conducted to determined apparent digestibility coefficients (ADC) of dry matter, crude protein, crude lipid, ash, energy and amino acids for twelve protein feed ingredients (i.e. fish meal, blood meal, meat and bone meal, poultry by-products meal, shrimp head meal, squid visceral meal, soybean meal, rapeseed meal, cottonseed meal, peanut meal, corn gluten meal and brewers yeast) for Pacific white shrimp (4.45±0.21 g). A reference diet and test diets (consisted of 70% reference diet and 30% of the raw feedstuff, except for blood meal with a ratio of 85%:15%) were used with Cr2O3 as external indicator. The results showed that the ADC of dry matter ranged from 48.61% to 86.98%, the ADC of energy ranged from 51.11% to 97.27%, the ADC of crude protein ranged from 55.74% to 92.35%, the ADC of crude lipid ranged from -0.23% to 92.64%, and the ADC of ash between 60.83% and 96.30%. The ADCs of total amino acids for fish meal, soybean meal and peanut meal were significantly higher than that for other meals (P<0.05), while corn gluten meal was significantly lower than that for other meals (P<0.05), and those feed ingredients were: soybean meal (92.67%) > peanut meal (90.61%) > fish meal (90.56%)> poultry by-products (89.81%) > shrimp head meal (88.82%) > brewers yeast (86.23%) > rapeseed meal (84.84%) > meat-bone meal (83.87%) > squid visceral meal (83.72%) > blood meal (77.25%) > cottonseed meal (72.82%) > corn gluten meal (59.15%).
Experiment 3: A feeding trial was conducted to evaluate the effect of five practical diets formulated on the same level of digestible crude protein, lysine, methionine and arginine on growth performance, body composition, feed utilization, digestive enzyme activity and values of biochemical parameters for the shrimp. The results showed that weight gain rate, survival, specific growth rate and protease activity in stomach in shrimp fed the diet at the fish meal inclusion level of 10% were significantly lower than that of shrimp fed the diets at the level of 15% or higher (P<0.05), while hepatosomatic indexes, blood urea nitrogen and lysozyme activities increased significantly (P<0.05). There were no differences in growth performance, body composition, digestive enzyme activity, blood urea nitrogen level, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase and lysozyme activities between the fish meal diets at the inclusion level of 15% or higher (P>0.05). Feed conversion rate and feeding rate increased, while protein efficiency ratio and apparent digestibility coefficients of dry matter, crude protein and crude lipid decreased with the reduction in dietary fish meal inclusion level.
Experiment 4: Six isonitrogenous (crude protein 41%) and isolipidic (crude lipid 8.5%) diets were formulated to contain 0%, 7%, 14%, 21%, 28% and 35% of peanut meal (PM), replacing 0%, 15%, 30%, 45%, 60% and 75% of fish meal (FM) protein respectively. Chromic oxide was used as the inert indicator for digestibility. The results showed that weight gain rate decreased significantly when dietary PM inclusion was raised above 21% (P<0.05). Protein efficiency ratio and survival rate decreased significantly but feed conversion rate increased significantly as PM inclusion level above 14% (P<0.05). Feeding rate in 21% PM inclusion group was significantly higher than that in 28% and 35% PM inclusion groups (P<0.05). Body crude protein and ash content significantly decreased, but body moisture significantly increased when dietary PM inclusion was higher than 14% (P<0.05). The apparent digestibility for dry matter, crude protein, crude lipids and ash decreased gradually as the level of PM replacement increased. Protease activities in stomach and hepatopancreas of 0~14% PM inclusion groups were significantly higher than in other groups (P<0.05). There was a similar trend as digestibility for the activities of superoxide dismutase (SOD), peroxidase (POD), acid phosphatase (ACP) and alkaline phosphatase (AKP) in serum, but ceruloplasmin (CP) content displayed adverse tendency. Body lipid and lysozyme activity was not affected by all treatments (P>0.05).
试验一:采用离体消化法研究了凡纳滨对虾的胃、中肠腺及肠道粗酶液对鱼粉、豆粕、菜粕和花生粕的离体消化率和酶解动力学。结果表明:凡纳滨对虾不同消化部位对干物质消化率的大小顺序为:胃>中肠腺>肠道,且鱼粉在胃部的消化率显著高于中肠腺和肠道(P<0.05),花生粕在肠道的消化率显著低于胃和中肠腺(P<0.05);对蛋白质消化率的大小顺序为:中肠腺>胃>肠道,且菜粕和花生粕在肠道的消化率显著低于胃和中肠腺(P<0.05),鱼粉在肠道和胃的消化率显著低于中肠腺(P<0.05);粗酶液对4种原料干物质的消化率高低依次为:豆粕50.78%、菜粕42.02%、花生粕39.67%、鱼粉36.50%;对粗蛋白的消化率高低依次为:花生粕60.40%、豆粕56.45%、鱼粉46.28%、菜粕43.28%。在0~4 h内,粗酶液酶解4种蛋白原料时氨基酸的生成量呈增加趋势;不同消化部位酶解产生的氨基酸总量不同,高低顺序依次为中肠腺(96.72 mg)>胃(31.28 mg)>肠道(27.58 mg);不同蛋白原料酶解时的氨基酸生成速率大小顺序为花生粕(3.9154 mg/h)>鱼粉(3.4774 mg/h)>豆粕(2.8316 mg/h)>菜粕(2.7404 mg/h)。
试验二:选取12种常见蛋白质饲料原料,按照70%基础饲料+30%试验原料(血粉按85%+15%)的原则配制饲料,以三氧化二铬(Cr2O3)作为外源指示剂,测定了凡纳滨对虾(4.45±0.21 g)对蛋白原料干物质、粗蛋白、粗脂肪、灰分、能量和氨基酸的表观消化率。结果表明:各种蛋白原料干物质表观消化率在48.61%~86.98%之间;粗蛋白表观消化率在55.74%~92.35%之间;粗脂肪的表观消化率在-0.23%~92.64%之间;灰分表观消化率在60.83%~96.30%之间;总能表观消化率在51.11%~97.27%之间。鱼粉、豆粕和花生粕的总氨基酸表观消化率分别为90.56%、92.67%和90.61%,显著高于其他蛋白原料(P<0.05);玉米蛋白粉的总氨基酸表观消化率最低,仅为59.15%,显著低于其他蛋白原料(P<0.05)。12种蛋白原料总氨基酸表观消化率高低顺序为:豆粕>花生粕>鱼粉>鸡肉粉>虾头粉>饲料酵母>菜粕>肉骨粉>乌贼内脏粉>血粉>棉粕>玉米蛋白粉。
试验三:以可消化粗蛋白和氨基酸为基础,设计可消化蛋白水平和可消化赖氨酸、蛋氨酸和精氨酸水平相同、但鱼粉添加比例不同的5种饲料配方(分别为30%、25%、20%、15%和10%),配制试验饲料,并比较这些饲料对凡纳滨对虾的饲喂效果。结果表明:随着饲料中鱼粉比例下降,饲料系数和摄食率逐渐升高,蛋白质效率、干物质、粗蛋白和粗脂肪表观消化率逐渐降低。饲料中鱼粉比例为10%时,对虾增重率、存活率、特定生长率和胃蛋白酶活性显著低于其他试验组(P<0.05),肝体指数、血清尿素氮含量和溶菌酶活性显著升高(P<0.05)。利用可消化蛋白及主要限制性氨基酸配制饲料,可使饲料中鱼粉添加比例降低至15%而不影响凡纳滨对虾的生长性能、饲料利用及营养状况。
试验四:以花生粕作为鱼粉蛋白替代原料,配制6种等氮等脂的试验饲料,其中花生粕的添加比例分别为0、7、14、21、28和35%,分别替代0、15、30、45、60和75%的鱼粉蛋白,同时以一种市售凡纳滨对虾饲料做参考饲料(RD)。饲料中添加Cr2O3作为外源指示剂用于消化率测定。结果表明:花生粕在饲料中比例增加到21%以上时,增重率显著低于对照组(P<0.05);花生粕比例增加到14%以上时,饲料效率、饲料蛋白质效率和存活率显著低于对照组(P<0.05);花生粕比例增加至35%时对虾摄食率显著低于对照组(P<0.05)。随花生粕添加比例增加,虾体水分含量呈升高趋势,粗蛋白和灰分含量则呈下降趋势,但粗脂肪含量变化不明显。饲料干物质、粗蛋白、粗脂肪和灰分的消化率随花生粕添加比例增加而降低,替代比例越高,消化率下降程度则越大。对虾胃蛋白酶和肝胰蛋白酶活性,血清中的超氧化物歧化酶、过氧化物酶、酸性磷酸酶和碱性磷酸酶活性变化趋势与消化率的变化趋势一致,铜蓝蛋白含量变化趋势则相反。花生粕不同替代比例对对虾血清溶菌酶活性没有显著影响(P>0.05)。
In this paper, the nutrient digestibility of different protein feed ingredients were evaluated using in vitro and in vivo methods for Pacific white shrimp, Litopenaeus vannamei. Based on digestibility of these protein feed ingredients, five practical diets were formulated to contain the same level of digestible crude protein and digestible amino acids for the shrimp, and their effect on growth performance, body compositions, feed utilization, digestive enzyme activity and biochemical parameters were assessed. In addition, the effect of replacement fish meal by peanut meal in practical diets on growth performance for the shrimp was also investigated. The results were as follows:
Experiment 1: The digestibility of four protein feed ingredients [fish meal (FM), soybean meal (SM), rapeseed meal (RM) and peanut meal (PM)] were determined using the in vitro method and kinetics of hydrolysis of digestive enzymes in stomach, hepatopancreas and intestine of Pacific white shrimp were studied. The results showed that the order of digestion of different digestive organs for dry matter was stomach > hepatopancreas > intestine, and stomach had significantly higher dry matter digestibility of FM than hepatopancreas and intestine (P<0.05), but intestine got significantly lower dry matter digestibility of PM than stomach and hepatopancreas (P<0.05). The order of digestion of different digestive organs for crude protein was hepatopancreas > stomach > intestine, and the digestibility of PM and RM in intestin were significantly lower than in stomach and hepatopancreas (P<0.05), but the digestibility of FM in hepatopancreas was significantly higher than in stomach and intestine (P<0.05). The order of proteins for digestibility of dry matter was SM (50.78%) > RM (42.02%) > PM (39.67%) > FM (36.50%). The order proteins for digestibility of crude protein was PM (60.40%) > SM (56.45%) > FM (46.28%) > RM (43.28%). The amino acids production of enzymolysis reaction was hepatopancreas (96.72 mg) > stomach (31.28 mg) > intestine (27.58 mg); the order of enzymolysis rate for amino acid was PM (3.9154 mg/h) > FM (3.4774 mg/h) > SM (2.8316 mg/h) > RM (2.7404 mg/h).
Experiment 2: A feeding trial was conducted to determined apparent digestibility coefficients (ADC) of dry matter, crude protein, crude lipid, ash, energy and amino acids for twelve protein feed ingredients (i.e. fish meal, blood meal, meat and bone meal, poultry by-products meal, shrimp head meal, squid visceral meal, soybean meal, rapeseed meal, cottonseed meal, peanut meal, corn gluten meal and brewers yeast) for Pacific white shrimp (4.45±0.21 g). A reference diet and test diets (consisted of 70% reference diet and 30% of the raw feedstuff, except for blood meal with a ratio of 85%:15%) were used with Cr2O3 as external indicator. The results showed that the ADC of dry matter ranged from 48.61% to 86.98%, the ADC of energy ranged from 51.11% to 97.27%, the ADC of crude protein ranged from 55.74% to 92.35%, the ADC of crude lipid ranged from -0.23% to 92.64%, and the ADC of ash between 60.83% and 96.30%. The ADCs of total amino acids for fish meal, soybean meal and peanut meal were significantly higher than that for other meals (P<0.05), while corn gluten meal was significantly lower than that for other meals (P<0.05), and those feed ingredients were: soybean meal (92.67%) > peanut meal (90.61%) > fish meal (90.56%)> poultry by-products (89.81%) > shrimp head meal (88.82%) > brewers yeast (86.23%) > rapeseed meal (84.84%) > meat-bone meal (83.87%) > squid visceral meal (83.72%) > blood meal (77.25%) > cottonseed meal (72.82%) > corn gluten meal (59.15%).
Experiment 3: A feeding trial was conducted to evaluate the effect of five practical diets formulated on the same level of digestible crude protein, lysine, methionine and arginine on growth performance, body composition, feed utilization, digestive enzyme activity and values of biochemical parameters for the shrimp. The results showed that weight gain rate, survival, specific growth rate and protease activity in stomach in shrimp fed the diet at the fish meal inclusion level of 10% were significantly lower than that of shrimp fed the diets at the level of 15% or higher (P<0.05), while hepatosomatic indexes, blood urea nitrogen and lysozyme activities increased significantly (P<0.05). There were no differences in growth performance, body composition, digestive enzyme activity, blood urea nitrogen level, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase and lysozyme activities between the fish meal diets at the inclusion level of 15% or higher (P>0.05). Feed conversion rate and feeding rate increased, while protein efficiency ratio and apparent digestibility coefficients of dry matter, crude protein and crude lipid decreased with the reduction in dietary fish meal inclusion level.
Experiment 4: Six isonitrogenous (crude protein 41%) and isolipidic (crude lipid 8.5%) diets were formulated to contain 0%, 7%, 14%, 21%, 28% and 35% of peanut meal (PM), replacing 0%, 15%, 30%, 45%, 60% and 75% of fish meal (FM) protein respectively. Chromic oxide was used as the inert indicator for digestibility. The results showed that weight gain rate decreased significantly when dietary PM inclusion was raised above 21% (P<0.05). Protein efficiency ratio and survival rate decreased significantly but feed conversion rate increased significantly as PM inclusion level above 14% (P<0.05). Feeding rate in 21% PM inclusion group was significantly higher than that in 28% and 35% PM inclusion groups (P<0.05). Body crude protein and ash content significantly decreased, but body moisture significantly increased when dietary PM inclusion was higher than 14% (P<0.05). The apparent digestibility for dry matter, crude protein, crude lipids and ash decreased gradually as the level of PM replacement increased. Protease activities in stomach and hepatopancreas of 0~14% PM inclusion groups were significantly higher than in other groups (P<0.05). There was a similar trend as digestibility for the activities of superoxide dismutase (SOD), peroxidase (POD), acid phosphatase (ACP) and alkaline phosphatase (AKP) in serum, but ceruloplasmin (CP) content displayed adverse tendency. Body lipid and lysozyme activity was not affected by all treatments (P>0.05).
引文
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