A Study to Explore the Minimum Dietary Fishmeal Content for Largemouth Bass (Micropterus salmoides)

NIU Jingyan; LIU Yu; HECTOR Victor; WANG Yan

doi:10.12418/CJAN2024.674

Chinese Journal of Animal Nutrition >

2024 , Vol. 36 >Issue 12: 7900 - 7914

DOI: https://doi.org/10.12418/CJAN2024.674

RESEARCH PAPER

A Study to Explore the Minimum Dietary Fishmeal Content for Largemouth Bass (Micropterus salmoides)

NIU Jingyan ^,¹^,² ,
LIU Yu ² ,
HECTOR Victor ² ,
WANG Yan ^,²^,^*

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¹ Institute of Technology, Jiaozuo Normal College, Jiaozuo 454000, China
² Ocean College, Zhejiang University, Zhoushan 316021, China

*professor, E-mail: ywang@zju.edu.cn

Received date: 2024-05-22

Online published: 2024-12-12

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Abstract

This study aimed to determine the minimum dietary fishmeal content for largemouth bass (Micropterus salmoides) with poultry by-product meal (PBM) as a single alternative ingredient, as well as the influences of salicylate supplementation on fishmeal replacement level. Two feeding experiments (1 and 2) were conducted. In experiment 1, a single-factor experimental design was used. The basal diet (F40) contained 40% fishmeal, and 80% and 90% of the fishmeal in F40 was substituted with PBM to formulate two low-fishmeal diets, F8 and F4, respectively. In experiment 2, a 2×2 double-factor experimental design was used. At first, two low-animal-protein diets, F8D and F4D, were formulated by replacing PBM in diets F8 and F4 partially with a blend of soy protein concentrate, cottonseed meal and dried porcine soluble (DPS), respectively. Thereafter, salicylate was added at 0.005% in diets F8D and F4D to formulate F8DS and F4DS. The proportion of animal protein to dietary protein was 63% in F40, F8 and F4, and it was reduced to 51% in F8D and F8DS, 49% in F4D and F4DS. Experiments 1 and 2 were carried out at the same time. Largemouth bass with initial body weight of (13.1±0.1) g were as experimental fish, and they were fed with experimental diets in a flowing-through rearing system for 10 weeks. The results showed as follows: in experiment 1, dietary fishmeal replacement level significantly affected feed conversion ratio (FCR) and retention efficiency of nitrogen (NRE), crude protein content of whole-fish, fish-in/fish-out ratio (FIFO) (P<0.05), but did not significantly affected final body weight (FBW), weight gain (WG), feed intake (FI), phosphorus retention efficiency (PRE), energy retention efficiency (ERE), condition factor (CF), hepatosomatic index (HSI), viscerosomatic index (VSI), contents of crude lipid, crude ash and gross energy, waste outputs of nitrogen (NW) and phosphorus (PW) (P>0.05). The WG declined by 1.5% and 7.9%, while the FBW declined by 1.8% and 9.4%, in fish fed F8 and F4 relative to fish fed F40. In experiment 2, salicylate added or not significantly affected FCR, NRE, HSI and NW (P<0.05), while dietary fishmeal content significantly affected crude lipid content of whole-fish and FIFO (P<0.05). FBW, WG, FI, PRE, ERE, CF and PW were not significantly affected by dietary fishmeal content, salicylate added or not and their interaction (P>0.05). The WG declined by 12.1%, 15.7%, 2.5% and 5.4%, while the FBW declined by 9.9%, 13.0%, 2.0% and 4.6%, in fish fed F8D, F4D, F8DS and F4DS relative to fish fed F40. This study reveals that reducing dietary fishmeal content for largemouth bass to 8% by using PBM as a single alternative protein source does not significantly affect growth, feed utilization efficiency and aquaculture waste outputs of largemouth bass, and reducing dietary fishmeal content to 4% results in apparently declined growth and feed utilization efficiency and significantly increases NW. Adding salicylate at 0.005% benefits to improve growth and NRE of largemouth bass fed the low-fishmeal diets. According to the results of this study, the application of fishmeal-free diets in commercial largemouth bass farming still face challenges.

Key words： poultry by-product meal; salicylate; minimum dietary fishmeal level; growth; feed utilization efficiency; aquaculture waste output

Cite this article

NIU Jingyan , LIU Yu , HECTOR Victor , WANG Yan . A Study to Explore the Minimum Dietary Fishmeal Content for Largemouth Bass (Micropterus salmoides)[J]. Chinese Journal of Animal Nutrition, 2024 , 36(12) : 7900 -7914 . DOI: 10.12418/CJAN2024.674

配合饲料应用是推动水产动物养殖产业快速发展的主要因素之一,而投饵养殖在水产动物养殖产量中所占的比例随着产业规模的扩大也逐年增加,2020年投饵养殖在世界水产动物养殖产量中所占的比例达到72%^[1],优质饲料蛋白质源短缺的问题也日益突出。鱼粉被公认为是优质的饲料蛋白质源。由于每年世界鱼粉产量的70%以上被用于生产水产饲料^[2],所以摆脱饲料生产对鱼粉的依赖性成为水产养殖,特别是鱼类和甲壳动物养殖产业能否实现可持续发展的关键^[3]。

大口黑鲈(Micropterus salmoides)为分布于北美内陆水域的凶猛肉食性鱼类,其在20世纪60年代后被进行人工养殖^[4],现已成为世界上重要的淡水鱼类养殖种类。2020年大口黑鲈养殖产量在淡水肉食性鱼类中和淡水鱼类中分别位居世界的第2位和第15位^[1],位居我国的第1位和第7位^[5]。大口黑鲈饲料最适蛋白质水平为48%~51%^[6],早期商业饲料鱼粉含量高达45%^[7]。2018年后围绕大口黑鲈饲料鱼粉替代问题开展了大量研究^{[8⇓⇓⇓⇓⇓⇓⇓⇓⇓⇓⇓⇓⇓-22]}。Wang等^[17]报道,利用鸡肉粉(PBM)、PBM与大豆浓缩蛋白(SPC)混合物或PBM与棉籽浓缩蛋白混合物为替代蛋白质源时,可将大口黑鲈饲料中鱼粉含量降低至8%;Mclean等^[21]报道,利用PBM与SPC混合物可完全替代大口黑鲈饲料中的鱼粉。由于饲料中鱼粉含量直接影响饲料生产成本以及养殖对海洋渔业资源的依赖性,故无鱼粉饲料能否在生产中应用对大口黑鲈养殖产业可持续性发展具有重要的意义,有必要进行进一步验证。

尽管过去40多年围绕水产动物饲料鱼粉替代问题开展了大量的研究,但在影响饲料中鱼粉替代水平的因素和机制方面仍存在许多知识空白点。一些研究表明饲料中的核苷酸和功能性氨基酸等可促进鱼类生长^{[23⇓⇓-26]},在低鱼粉饲料中添加这类具有促生长作用的成分作为功能性添加剂有益于改善因饲料鱼粉含量降低所引发的鱼类生长缓慢的问题^[27]。肠膜蛋白(DPS)是猪小肠黏膜酶解、干燥后的产物,其富含小肽、游离态氨基酸、酶、激素、细胞因子和生物胺等成分^[28],被广泛用于禽类和猪饲粮中^{[28⇓⇓⇓-32]}。在水产饲料中,DPS被作为鱼粉的替代蛋白质源,但不同研究所得出的结论^{[33⇓⇓⇓-37]}并不一致。水杨酸盐具有清除机体内自由基、抗氧化的作用^[38],其对鱼类生长的影响尚未见报道。本研究评价了利用PBM替代大口黑鲈饲料中不同水平的鱼粉以及在此基础上利用DPS与植物蛋白质的混合物替代低鱼粉饲料中PBM对鱼摄食、生长和饲料利用的影响,并初步探讨了在低鱼粉饲料中添加水杨酸盐的作用,旨在探索大口黑鲈饲料中可实现的最低鱼粉含量以及降低饲料鱼粉含量的途径。

1 材料与方法

1.1 伦理审查

动物试验已报浙江大学试验动物福利伦理审查委员会审查并获得批准(批准号:ZJU20240576)。

1.2 试验鱼和流水养殖系统

大口黑鲈饲养试验在浙江省杭州市余杭区恒泽生态农业科技有限公司进行。所用鱼种由当地一家淡水鱼苗种场孵化培育,用活水车运至试验地点。试验鱼抵达后先暂养在多个容积为300 L的聚乙烯水槽内,暂养期间投喂由浙江大学水域生态系统与水产养殖试验室提供配方,由德清县鸿利生物科技有限公司加工生产的配合饲料(饲料中粗蛋白质、粗脂肪和粗灰分含量分别为50%、9%和13%)。所用的流水养殖试验系统(以下简称流水系统)安装在一个可移动的帐篷(6 m×10 m)内,包括进水系统(由1台0.75 kW潜水泵、进水管道和对应每个水槽的进水阀门组成)、48个试验水槽(直径80 cm、高80 cm、容积300 L的聚乙烯水槽)、增氧系统(由1台0.40 kW充气泵、输气管道和对应每个水槽的气阀、气管与气石组成)和排水系统(由对应每个水槽的排水管道、放水阀门和总排水管组成)。每个试验水槽可独立进水、排水和充气。移动帐篷搭建在一口池塘[面积为20亩(1亩≈667 m²),放养少量鲢、鳙,不投饵,透明度>60 cm]边,从池塘表层下50 cm处抽水作为流水系统的水源。试验前2周,挑选1 050尾个体大小接近、外表无伤的试验鱼移入流水系统内驯养(驯养的试验鱼放入21个水槽中,密度为每个水槽50尾)。

1.3 试验设计和饲料

共进行2个饲养试验。试验1采用单因素试验设计,基础饲料(F40)含40%的鱼粉,利用PBM分别替代F40中鱼粉的80%和90%构建饲料F8和F4,F40、F8和F4中动物蛋白质占饲料蛋白质的比例均为63%。试验2采用2×2双因素试验设计,首先利用SPC、棉籽粕和DPS组成的混合物替代F8和F4中的部分PBM构建饲料F8D和F4D(F8D和F4D中动物蛋白质占饲料蛋白质的比例分别为51%和49%),再在F8D和F4D中分别添加0.005%的水杨酸盐构建饲料F8DS和F4DS。试验所用的水杨酸盐(纯度≥99.5%)购自上海国药集团化学试剂有限公司;鱼粉(超级蒸汽鱼粉,秘鲁)、PBM(宠物级,美国)、血球蛋白粉、豆粕、棉籽粕和面粉(高筋)、DPS(商业产品)等主要饲料原料的营养组成见表1。

表1 主要饲料原料的营养组成(风干基础)

Table 1 Nutritional composition of main feed ingredients (air-dry basis) %

项目 Items	干物质 Dry matter	粗蛋白质 Crude protein	粗脂肪 Crude lipid	粗灰分 Crude ash
鱼粉Fishmeal	89.4	64.6	9.7	14.9
鸡肉粉Poultry by-product meal	93.1	65.2	9.3	18.4
预混合蛋白Protein premix	90.7	60.0	1.0	6.8
血球蛋白粉Blood cell protein meal	91.3	92.3	0.2	3.9
豆粕Soybean meal	90.3	49.1	1.3	6.1
棉籽粕Cottonseed meal	90.9	61.1	0.6	7.8
面粉Wheat flour	85.9	17.7	2.1	1.1
肠膜蛋白Dried porcine soluble	92.2	52.1	8.8	21.0

	数据为2次测定结果的平均值。预混合蛋白为一种商业产品,其成分包括血球蛋白粉、豆粕和棉籽粕,表2同。
	Data are mean values of two measured results. The protein premix is a commercial product consisting of blood cell protein meal, soybean meal and cottonseed meal, the same as Table 2.

利用锤片式粉碎机将饲料原料粉碎,直至其全部通过40目筛。按试验饲料制作流程^[6,8]依次称量配方中的各个原料,逐级混合、加水搅拌,最后利用单螺杆膨化饲料机挤压制成直径约为3mm、长度约为5 mm的沉性颗粒。待试验饲料在室温下风干后,将其收集在聚乙烯自封袋内并保存在冰箱(-20 ℃)中^[11,17]。试验饲料组成及营养水平见表2。DPS和部分试验饲料(F40、F8和F4)的氨基酸组成见表3。

表2 试验饲料组成及营养水平(风干基础)

Table 2 Composition and nutrient levels of experimental diets (air-dry basis) %

项目 Items	饲料Diets
项目 Items	F40	F8	F4	F8D	F4D	F8DS	F4DS
原料Ingredients
鱼粉Fishmeal	40.0	8.0	4.0	8.0	4.0	8.0	4.0
鸡肉粉Poultry by-product meal		30.4	34.2	22.8	22.8	22.8	22.8
大豆浓缩蛋白Soy protein concentrate				6.0	8.5	6.0	8.5
棉籽粕Cottonseed meal	4.0	4.0	4.0	7.0	6.0	7.0	6.0
肠膜蛋白Dried porcine soluble				0.5	0.5	0.5	0.5
血球蛋白粉Blood cell protein meal	3.2	3.2	3.2	2.7	4.4	2.7	4.4
豆粕Soybean meal	8.0	8.0	8.0	8.0	8.0	8.0	8.0
预混合蛋白Protein premix	20.0	20.0	20.0	20.0	20.0	20.0	20.0
面粉Wheat flour	13.5	13.5	13.5	13.5	13.5	13.5	13.5
木薯淀粉Cassava starch	0.1	0.1	0.1	0.1	0.1	0.1	0.1
膨润土Bentonite		2.8	3.2	0.6	1.0	0.6	1.0
氯化胆碱Choline chloride	0.2	0.2	0.2	0.2	0.2	0.2	0.2
磷酸二氢钙Ca(H₂PO₄)₂	2.0	2.0	2.0	2.0	2.0	2.0	2.0
DL-蛋氨酸DL-methionine	0.2	0.2	0.2	0.2	0.2	0.2	0.2
L-赖氨酸L-lysine	0.5	0.5	0.5	0.5	0.5	0.5	0.5
维生素和矿物预混料 Vitamin and mineral premix¹⁾	1.5	1.5	1.5	1.5	1.5	1.5	1.5
鱼油Fish oil	6.8	5.6	5.4	6.4	6.8	6.4	6.8
合计Total	100.0	100.0	100.0	100.0	100.0	100.0	100.0
水杨酸盐Salicylate						0.005	0.005
营养水平Nutrient levels²⁾
干物质Dry matter	93.5	93.2	93.5	93.8	94.1	93.8	94.1
粗蛋白质Crude protein	51.9	51.1	50.5	51.3	50.5	51.3	50.5
粗脂肪Crude lipid	11.0	10.3	10.2	10.5	10.0	10.5	10.0
粗灰分Crude ash	12.7	11.0	11.2	11.0	11.1	11.0	11.1
总能Gross energy/(MJ/kg)	19.5	19.3	19.1	19.4	19.3	19.4	19.3
磷Phosphorus	1.23	1.07	1.14	1.12	1.09	1.12	1.09

1)维生素和矿物质预混料组成详见文献[20]。Composition of the vitamin and mineral premix is described in the literature[20].

2)营养水平数据为2次测定结果的平均值。The data of nutrient levels are mean values of two measured results.

表3 肠膜蛋白和部分试验饲料的氨基酸组成(风干基础)

Table 3 Amino acid composition of dried porcine soluble and some experimental diets (air-dry basis) %

氨基酸 Amino acids	肠膜蛋白 Dried porcine soluble	饲料Diets
氨基酸 Amino acids	肠膜蛋白 Dried porcine soluble	F40	F8	F4
蛋氨酸Methionine	0.95	0.91	0.79	0.74
赖氨酸Lysine	3.12	4.21	3.82	3.65
精氨酸Arginine	2.29	3.28	3.53	3.48
亮氨酸Leucine	3.75	4.39	4.34	4.22
异亮氨酸Isoleucine	2.15	2.21	2.12	2.06
缬氨酸Valine	2.63	3.00	2.95	2.87
组氨酸Histidine	1.09	2.01	1.63	1.53
酪氨酸Tyrosine	1.45	1.33	1.32	1.25
苏氨酸Threonine	2.43	2.18	2.09	2.04
丝氨酸Serine	2.21	2.33	2.38	2.35
甘氨酸Glycine	3.26	2.97	3.84	3.89
丙氨酸Alanine	1.85	3.22	3.29	3.22
天冬氨酸Aspartic acid	4.30	5.40	5.30	5.15
脯氨酸Proline	2.61	2.28	2.76	2.76
谷氨酸Glutamic acid	6.94	8.48	8.65	8.47
苯丙氨酸Phenylalanine	1.98	2.57	2.64	2.52

	数据为2次测定结果的平均值。
	Data are mean values of two measured results.

1.4 饲养试验

试验1和试验2同时进行,试验开始前将在流水系统中驯养的大口黑鲈停喂24 h。每次挑选30尾个体大小相近、体表无伤的大口黑鲈,群体称重后随机放入1个试验水槽内,每种饲料设3个重复(水槽),共放养21个水槽(n=21),试验鱼初始体质量为(13.1±0.1) g。放养完成后从剩余的鱼中随机捕捞15尾鱼(分为3组,每组5尾),测定每尾鱼的体长、体质量、肝脏质量和内脏团质量,将鱼放入聚乙烯自封袋内并保存在冰箱(-20 ℃)中作为分析试验开始时鱼体组成的样品。

饲养试验时间为10周。每天06:00和18:00按饱食量投喂试验饲料。每天14:00采用虹吸方法清除水槽底部的沉积物。试验水槽中连续流水(每个水槽水交换速率为2.0~3.0 L/min)和充气。采用自然光照,光∶暗时间为14 h∶10 h。每天早、晚分别测量试验水槽内的水温(试验期间变化范围为27.2~32.6 ℃);每周测定1次养殖水体的pH以及溶氧、氨态氮和亚硝酸态氮浓度(试验期间水体pH为7.20~8.41,溶氧浓度为3.88~8.72 mg/L,氨态氮浓度为0.014~0.033 mg/L,亚硝酸态氮浓度为0.002~0.079 mg/L)。

饲养试验结束前先将鱼停喂24 h,然后将每个水槽内的鱼依次捕出、麻醉(丁香酚∶无水乙醇=1∶9)并群体称重。从每个水槽取5尾鱼,单尾测量其体长、体质量、肝脏质量和内脏团质量,然后将其放入聚乙烯自封袋并保存在冰箱(-20 ℃)中作为分析鱼体组成的样品。

1.5 营养成分分析

将试验鱼样品从冰箱中取出,在室温下化冻、称重,在高压蒸汽灭菌锅(120 ℃)内将鱼蒸煮20 min,再用食品粉碎机将其打成鱼浆,放入烘箱(105 ℃)内烘干。用高速粉碎机将烘干的试验鱼、饲料和饲料原料粉碎直至通过80目筛。试验鱼和试验饲料的水分(或干物质)、粗蛋白质、粗脂肪、粗灰分、总能、磷和氨基酸含量以及饲料原料的干物质、粗蛋白质、粗脂肪和粗灰分含量参照AOAC(1995)^[39]的方法测定。其中,使用8400型全自动凯氏氮分析仪(Foss,瑞典)测定粗蛋白质含量;用SZF-06A型脂肪抽提仪(上海新嘉电子有限公司,中国)测定粗脂肪含量;使用6200型氧弹仪(Parr,美国)测定总能含量;使用433型氨基酸分析仪(Sykam,德国)测定氨基酸含量。

1.6 计算公式

存活率(SR)、鱼体增重(WG)、摄食率(FI)、饲料系数(FCR)、氮贮积效率(NRE)、磷贮积效率(PRE)、能量贮积效率(ERE)、氨基酸贮积效率(ARE)、肥满度(CF)、肝体指数(HSI)、脏体指数(VSI)、氮废物排放量(NW)和磷废物排放量(PW)以及单位鱼产量的鱼粉消耗量(FIFO)分别按下列公式计算:

SR(%)=100×N_t/N₀;

WG(g)=W_t/N_t-W₀/N₀;

FI(%/d)=100×I/[(W₀+W_t)/2×t];

FCR=I/(W_t-W₀+W_d);

NRE(%)=100×[(W_t/N_t×C_Nt)-(W₀/N₀×C_N0)]/[2×I/(N_t+N₀)×C_Nf];

PRE(%)=100×[(W_t/N_t×C_Pt)-(W₀/N₀×C_P0)]/[2×I/(N_t+N₀)×C_Pf];

ERE(%)=100×[(W_t/N_t×C_Et)-(W₀/N₀×C_E0)]/[2×I/(N_t+N₀)×C_Ef];

ARE(%)=100×[(W_t/N_t×C_At)-(W₀/N₀×C_A0)]/[2×I/(N_t+N₀)×C_Af];

CF(g/cm³)=100×W_s/

L s 3

;

HSI(%)=100×W_L/W_s;

VSI(%)=100×W_V/W_s;

NW(g N/kg鱼产量)=10×[I/(N_t+N₀)×2×C_Nf]×(1-NRE/100)/(W_t/N_t-W₀/N₀);

PW(g P/kg鱼产量)=10×[I/(N_t+N₀)×2×C_Pf]×(1-PRE/100)/(W_t/N_t-W₀/N₀);

FIFO(g鱼粉/g鱼产量)=I×FL/(W_t×DMF_t-W₀×DMF₀)。

式中:N₀(尾)和N_t(尾)分别为试验开始和结束时每个水槽内的试验鱼尾数;W₀(g)和W_t(g)分别为试验开始和结束时每个水槽内的试验鱼体质量;I(g)为试验期间每个水槽投喂的饲料量;t(d)为试验时间;W_d(g)为试验期间每个水槽内死鱼的体质量;C_N0(%)、C_P0(%)、C_E0(MJ/kg)和C_A0(%)分别为试验开始时鱼体的氮、磷、总能和氨基酸含量;C_Nt(%)、C_Pt(%)、C_Et(MJ/kg)和C_At(%)分别为试验结束时鱼体的氮、磷、总能和氨基酸含量;C_Nf(%)、C_Pf(%)、C_Ef(MJ/kg)和C_Af(%)分别为试验饲料的氮、磷、总能和氨基酸含量;W_s(g)、L_s(cm)、W_L(g)和W_V(g)分别为试验结束时采样鱼的体质量、体长、肝脏质量和内脏团质量;F_L(%)为试验饲料中鱼粉的干物质含量;DMF₀(%)和DMF_t(%)分别为试验开始和结束时鱼体的干物质含量。

1.7 数据统计分析

利用SPSS 25.0进行统计分析,当P<0.05时判定为差异显著。采用单因素方差分析检验饲料中鱼粉替代水平对试验鱼的终末体质量(FBW)、WG、FI、FCR、NRE、PRE、ERE、ARE、CF、HSI、VSI、NW、PW、FIFO和体组成的影响,若处理效应显著则进一步用邓肯氏检验(Duncan’s test)比较鱼粉含量不同的饲料(F40、F8和F4)间的差异显著性。采用双因素方差分析检验饲料鱼粉含量、添加水杨酸盐与否及二者交互作用对上述指标的影响(涉及F8D、F4D、F8DS和F4DS),若某一指标受主效应显著影响而交互作用不显著,则进一步用Duncan’s test比较鱼粉含量之间或水杨酸盐添加水平之间的差异。用邓尼特检验(Dunnett test)分别比较F40与F8D、F4D、F8DS和F4DS之间在FBW、WG、FI、FCR、NRE、PRE、ERE、CF、HSI、VSI、体组成、NW、PW以及FIFO方面的差异。

2 结果

2.1 大口黑鲈的存活、生长、摄食和饲料利用效率

试验1的测定结果(表4和表5)显示:试验过程中摄食不同饲料的大口黑鲈均未出现死亡现象;饲料中鱼粉替代水平显著影响FCR和NRE(P<0.05,但对WG和FBW无显著影响(P>0.05);摄食F4的大口黑鲈的FCR显著高于摄食F40的大口黑鲈(P<0.05),但NRE显著低于后者(P<0.05)。大口黑鲈的ARE表现为甘氨酸和蛋氨酸的贮积效率高于其他氨基酸,其次为丙氨酸和苏氨酸的贮积效率(仅摄食F40时脯氨酸的贮积效率高于苏氨酸的贮积效率),再次为异亮氨酸、脯氨酸、天冬氨酸、赖氨酸、精氨酸、酪氨酸、谷氨酸、丝氨酸、亮氨酸和缬氨酸的贮积效率,苯丙氨酸和组氨酸的贮积效率在所有氨基酸中最低;饲料鱼粉替代水平显著影响甘氨酸和脯氨酸的贮积效率(P<0.05),而对蛋氨酸、丙氨酸、苏氨酸、天冬氨酸、异亮氨酸、丝氨酸、酪氨酸、赖氨酸、亮氨酸、谷氨酸、缬氨酸、精氨酸、组氨酸、苯丙氨酸的贮积效率无显著影响(P>0.05);摄食F40的大口黑鲈的甘氨酸和脯氨酸贮积效率显著高于摄食F8和F4的大口黑鲈(P<0.05)。

表4 大口黑鲈的存活、生长、摄食和饲料利用效率

Table 4 Survival, growth, feed intake and feed utilization efficiency of largemouth bass

项目 Items	终末体质量 FBW/(g/尾)	增重 WG/g	摄食率 FI/(%/d)	饲料系数 FCR	氮贮积效率 NRE/%	磷贮积效率 PRE/%	能量贮积效率 ERE/%	存活率 SR/%
试验1 Experiment 1
F40	74.47±1.50	61.32±1.45	2.15±0.19	1.02±0.09^a	32.88±2.68^b	62.25±3.71	32.81±2.37	100
F8	73.42±2.22	60.22±2.27	2.32±0.08	1.12±0.04^ab	29.52±1.21^ab	64.29±5.13	30.82±0.33	100
F4	68.65±4.93	55.51±4.93	2.48±0.17	1.22±0.07^b	27.64±1.86^a	53.59±5.13	28.27±2.35	100
P值P-value	0.142	0.140	0.097	0.042	0.048	0.067	0.074
试验2 Experiment 2
F8D	67.08±2.36	53.95±2.36	2.44±0.17	1.22±0.09^*	27.71±1.94^*	57.04±4.54	28.07±3.98	100
F4D	64.77±1.39	51.71±1.60	2.48±0.03	1.24±0.02^*	26.84±0.19^*	53.39±2.70	27.52±0.36	99
F8DS	72.96±2.81	59.85±2.82	2.30±0.09	1.11±0.06	30.37±1.49	57.45±6.44	31.42±1.41	100
F4DS	71.12±4.42	58.02±4.40	2.34±0.10	1.15±0.06	30.61±1.26	60.93±6.05	30.28±1.77	99
饲料鱼粉含量Dietary fish meal content
8%鱼粉含量8% fish meal content	70.02±3.97	56.90±3.98	2.37±0.15	1.16±0.09	29.06±2.33	57.24±4.99	29.75±3.24	100
4%鱼粉含量4% fish meal content	69.55±5.17	56.47±5.14	2.41±0.10	1.19±0.06	28.62±2.34	57.16±5.88	28.90±1.89	99
水杨酸盐添加与否Salicylate added or not
不添加水杨酸盐Non-salicylate addition	67.53±4.30	54.44±4.28	2.46±0.11	1.23±0.06^β	27.28±1.32^α	55.21±3.89	27.80±2.54	99.5
添加水杨酸盐Salicylate addition	72.04±3.46	58.94±3.45	2.32±0.09	1.13±0.06^α	30.39±1.61^β	59.19±5.91	30.85±1.56	99.5
P值P-value
饲料鱼粉含量Dietary fish meal content	0.849	0.870	0.566	0.473	0.637	0.979	0.542
水杨酸盐添加与否Salicylate added or not	0.104	0.106	0.068	0.019	0.010	0.217	0.050
交互作用Interaction	0.589	0.581	0.855	0.980	0.671	0.264	0.829

除存活率表示为平均值外,其他数据均表示为平均值±标准差(n=3)。上标小写字母表示对试验1中F40、F8和F4进行单因素方差分析后的邓肯氏检验结果,大写字母和希腊字母表示对试验2中饲料鱼粉含量、水杨酸盐添加与否及其交互作用进行双因素方差分析后分别针对饲料鱼粉含量和水杨酸盐添加水平进行的邓肯氏检验结果,星号表示对F40与试验2中低鱼粉饲料(F8D、F4D、F8DS和F4DS)进行的邓尼特检验结果。同列数据上标字母不同者表示存在显著差异(P<0.05),上标星号者表示与F40存在显著差异(P<0.05)。表6和表7同。

Data are presented as mean±SD (n=3) except SR is presented as mean value. The small letters represent the results of Duncan’s test between F40, F8 and F4 following one-way ANOVA in experiment 1. The capital letters and Greek letters represent the results of Duncan's test either between the treatments with different fish meal contents or between the treatments with or without salicylate addition following two-way ANOVA in experiment 2. The asterisk represents the results of Dunnett test between F40 and low fish meal diets (F8D, F4D, F8DS, F4DS) in experiment 2. The data with different letters within the same column are significantly different (P<0.05), while the data with asterisk are significantly different with F40. The same as Table 6 and Table 7.

表5 大口黑鲈对饲料中氨基酸的贮积效率

Table 5 Retention efficiency of amino acids in diets for largemouth bass %

氨基酸 Amino acids	饲料Diets			P值 P-value
氨基酸 Amino acids	F40	F8	F4	P值 P-value
蛋氨酸Methionine	43.57±3.54	45.25±1.15	47.18±4.88	0.499
甘氨酸Glycine	53.68±4.37^b	37.96±0.97^a	36.03±3.73^a	0.001
丙氨酸Alanine	38.49±3.13	34.15±0.87	33.90±3.51	0.151
苏氨酸Threonine	33.77±2.74	31.93±0.81	31.93±3.30	0.612
天冬氨酸Aspartic acid	31.03±2.52	28.61±0.72	28.95±2.99	0.429
脯氨酸Proline	37.36±3.04^b	28.17±0.72^a	27.08±2.80^a	0.004
异亮氨酸Isoleucine	33.12±2.69	31.14±0.79	31.73±3.28	0.630
丝氨酸Serine	29.90±2.43	26.30±0.67	25.84±2.67	0.108
酪氨酸Tyrosine	29.97±2.43	27.93±0.71	28.82±2.98	0.571
赖氨酸Lysine	29.22±2.37	29.11±0.74	30.18±3.12	0.825
亮氨酸Leucine	28.26±2.29	25.76±0.65	26.03±2.69	0.337
谷氨酸Glutamic acid	29.57±2.40	26.12±0.66	26.07±2.70	0.146
缬氨酸Valine	27.75±2.25	25.32±0.64	25.86±2.67	0.375
精氨酸Arginine	31.56±2.57	26.67±0.68	26.56±2.75	0.052
苯丙氨酸Phenylalanine	26.86±2.18	23.36±0.59	24.24±2.51	0.152
组氨酸Histidine	18.36±1.49	20.24±0.51	21.65±2.24	0.110
氨基酸总量Total AA	31.62±2.57	26.75±0.68	26.63±2.81	0.050

	数据表示为平均值±标准差(n=3)。字母表示试验1中对F40、F8和F4进行单因素方差分析后的邓肯氏检验结果,同行数据上标不同字母者表示存在显著差异(P<0.05)。
	Data are presented as mean±SD (n=3). The superscripts represent the results of Duncan’s test on F40, F8 and F4 following one-way ANOVA in experiment 1, and the data with different letters within the same row is significantly different (P<0.05).

试验2的测定结果(表4)显示:试验结束时摄食不同饲料的大口黑鲈的存活率为99%~100%;水杨酸盐添加与否显著影响FCR和NRE(P<0.05),而饲料鱼粉含量、水杨酸盐添加与否及二者的交互作用对WG、FBW、FI、PRE和ERE无显著影响(P>0.05);摄食添加水杨酸盐饲料(F8DS和F4DS)的大口黑鲈的FCR显著低于摄食未添加水杨酸盐的饲料(F8D和F4D)的大口黑鲈(P<0.05),NRE显著高于后者(P<0.05);与摄食F40的大口黑鲈相比,摄食F8DS和F4DS的大口黑鲈的FBW、WG、FI、FCR、PRE和ERE与前者无显著差异(P>0.05),但摄食F8D和F4D的大口黑鲈的FCR显著高于前者(P<0.05),其NRE显著低于前者(P<0.05)。

2.2 大口黑鲈的形态和体组成

试验1中,饲料中鱼粉替代水平显著影响全鱼粗蛋白质含量(P<0.05),但对CF、HSI、VSI以及全鱼粗脂肪、粗灰分和总能含量无显著影响(P>0.05);摄食F40的大口黑鲈全鱼粗蛋白质含量显著高于摄食F8和F4的大口黑鲈(P<0.05)(表6)。

表6 大口黑鲈的肥满度、肝体指数、脏体指数和体组成

Table 6 CF, HSI, VSI and body composition of largemouth bass (n=3)

项目 Items	肥满度 CF/(g/cm³)	肝体指数 HSI/%	脏体指数 VSI/%	体组成Body composition
项目 Items	肥满度 CF/(g/cm³)	肝体指数 HSI/%	脏体指数 VSI/%	水分 Moisture/%	粗蛋白质 Crude protein/%	粗脂肪 Crude lipid/%	粗灰分 Crude ash/%	磷 Phosphorus/ %	总能 Gross energy/ (MJ/kg)
初始Initial	2.49±0.39	2.07±0.73	7.81±1.40	75.21±0.33	15.52±0.11	5.04±0.24	3.70±0.12	0.64±0.05	5.52±0.13
试验1 Experiment 1
F40	1.99±0.04	2.39±0.11	6.87±0.20	71.02±0.37	17.10±0.20^b	7.09±0.28	4.30±0.05	0.76±0.02	6.36±0.09
F8	2.00±0.04	2.40±0.40	6.88±0.48	71.68±0.27	16.62±0.10^a	6.92±0.18	4.21±0.12	0.76±0.04	6.46±0.04
F4	1.98±0.04	1.89±0.13	6.68±0.19	71.46±0.29	16.74±0.06^a	7.08±0.31	4.23±0.05	0.74±0.02	6.40±0.12
P值P-value	0.803	0.081	0.713	0.108	0.014	0.775	0.296	0.604	0.427
试验2 Experiment 2
F8D	1.96±0.03	2.38±0.14	6.75±0.26	71.23±0.18	16.92±0.09	7.00±0.08	4.33±0.12	0.75±0.04	6.31±0.38
F4D	2.02±0.02	1.93±0.17^*	6.17±0.28^*	71.74±0.24	16.62±0.09^*	6.95±0.22	4.13±0.04	0.72±0.03	6.49±0.10
F8DS	1.96±0.02	2.20±0.07	6.62±0.12	70.99±0.71	16.82±0.28	7.41±0.24	4.28±0.14	0.76±0.06	6.51±0.05
F4DS	1.95±0.06	2.62±0.32^*	7.08±0.45^*	71.69±0.19^*	16.76±0.04^*	6.70±0.21	4.28±0.10	0.75±0.08	6.51±0.11
饲料鱼粉含量Dietary fish meal content
8%鱼粉含量8% fish meal content	1.96±0.02	2.29±0.14	6.69±0.20	71.11±0.48^B	16.87±0.20	7.21±0.28^B	4.30±0.12	0.75±0.05	6.41±0.21
4%鱼粉含量4% fish meal content	1.98±0.05	2.27±0.41	6.62±0.60	71.72±0.20^A	16.69±0.10	6.82±0.24^A	4.20±0.10	0.74±0.05	6.50±0.09
水杨酸盐添加与否Salicylate added or not
不添加水杨酸盐Non-salicylate addition	1.99±0.04	2.15±0.28^α	6.46±0.40	71.48±0.34	16.77±0.18	6.97±0.15	4.23±0.14	0.74±0.03	6.40±0.21
添加水杨酸盐Salicylate addition	1.95±0.04	2.41±0.27^β	6.85±0.39	71.34±0.60	16.79±0.18	7.05±0.44	4.28±0.11	0.76±0.06	6.51±0.08
P值P-value
饲料鱼粉含量Dietary fish meal content	0.301	0.832	0.727	0.033	0.104	0.009	0.233	0.653	0.346
水杨酸盐添加与否Salicylate added or not	0.112	0.024	0.057	0.622	0.872	0.421	0.461	0.584	0.234
交互作用Interaction	0.086	0.002	0.018	0.723	0.429	0.022	0.108	0.726	0.346

试验2中,饲料鱼粉含量显著影响全鱼水分和粗脂肪含量(P<0.05),水杨酸盐添加与否显著影响HSI(P<0.05),饲料鱼粉含量与水杨酸盐添加与否对HSI、VSI和全鱼粗脂肪含量产生显著的交互影响(P<0.05);与摄食F40的大口黑鲈相比,摄食F4D的大口黑鲈HSI、VSI和全鱼粗蛋白质含量显著下降(P<0.05),摄食F4DS的大口黑鲈HSI、VSI和全鱼水分含量显著增加(P<0.05),但全鱼粗蛋白质含量显著下降(P<0.05)(表6)。

2.3 大口黑鲈养殖的鱼粉依赖性和废物排放量

试验1中,饲料中鱼粉替代水平显著影响FIFO(P<0.05),但对NW和PW无显著影响(P>0.05);随饲料中鱼粉替代水平的增加,大口黑鲈的FIFO呈现显著下降(P<0.05)(表7)。

表7 大口黑鲈养殖的鱼粉依赖性和废物排放量

Table 7 Fish meal reliance and waste outputs of largemouth bass farming (n=3)

项目 Items	单位鱼产量的鱼粉消耗量 FIFO/(g鱼粉/g鱼产量)	氮废物排放量 NW/(g N/kg鱼产量)	磷废物排放量 PW/(g P/kg鱼产量)
试验1 Experiment 1
F40	1.30±0.09^c	57.39±7.30	4.79±0.88
F8	0.30±0.01^b	64.43±1.90	4.30±0.67
F4	0.16±0.01^a	71.65±6.13	6.53±1.09
P值P-value	<0.001	0.063	0.057
试验2 Experiment 2
F8D	0.31±0.03^*	72.47±7.20^*	5.84±1.13
F4D	0.17±0.00^*	74.37±1.15^*	6.42±0.43
F8DS	0.26±0.02^*	62.84±5.21	5.77±1.08
F4DS	0.14±0.01^*	63.34±5.18	5.01±0.81
饲料鱼粉含量Dietary fish meal content
8%鱼粉含量8% fish meal content	0.27±0.02^B	67.42±7.86	5.84±0.93
4%鱼粉含量4% fish meal content	0.15±0.01^A	68.85±7.07	5.72±0.97
水杨酸盐添加与否Salicylate added or not
不添加水杨酸盐Non-salicylate addition	0.22±0.08	73.42±4.73^β	6.17±0.74
添加水杨酸盐Salicylate addition	0.20±0.07	63.09±4.65^α	5.39±0.95
P值P-value
饲料鱼粉含量Dietary fish meal content	<0.001	0.661	0.809
水杨酸盐添加与否Salicylate added or not	0.067	0.011	0.157
交互作用Interaction	0.320	0.720	0.244

试验2中,饲料鱼粉含量显著影响FIFO(P<0.05),而水杨酸盐添加与否显著影响NW(P<0.05);摄食含8%鱼粉的饲料(F8D和F8DS)的大口黑鲈的FIFO显著高于摄食含4%鱼粉饲料(F4D和F4DS)的大口黑鲈(P<0.05),摄食添加水杨酸盐饲料(F8DS和F4DS)的大口黑鲈的NW显著低于摄食不添加水杨酸盐饲料(F8D和F4D)的大口黑鲈(P<0.05);与摄食F40的大口黑鲈相比,摄食F8D、F4D、F8DS和F4DS的大口黑鲈的FIFO显著降低(P<0.05),摄食F8D和F4D的大口黑鲈的NW显著增加(P<0.05)(表7)。

3 讨论

3.1 以PBM为替代蛋白质源时大口黑鲈饲料的最低鱼粉含量

已发表的文献中,所采用的评价水产饲料鱼粉替代水平的标准尚不统一。绝大多数研究中以“不降低试验动物生长性能的前提下基础饲料中鱼粉可被替代的最大百分比”作为饲料鱼粉替代水平^[40-41]。这一指标反映的是能够从基础饲料中替代多少鱼粉,而不是能够将基础饲料中的鱼粉含量降低到何种程度,因此,当所用的基础饲料配方不合理(如鱼粉含量偏高)时得出的结果没有实际意义^[42]。Wang等^[42⇓-44]建议用“在不降低试验动物最大生长性能的前提下通过添加替代原料能够实现的饲料最低鱼粉含量”来评价饲料鱼粉替代水平。Naylor等^[3]用饲料最低鱼粉含量的变化评价了1997—2017年世界水产饲料鱼粉替代技术的进步。

有关大口黑鲈饲料鱼粉含量究竟可降低到何种程度仍无确切的答案。Tidwell等^[45]报道,利用PBM可完全替代大口黑鲈饲料中的鱼粉,但其试验饲料蛋白质水平(38%)明显低于大口黑鲈的饲料蛋白质需求量^[6],饲料蛋白质水平偏低掩盖了饲料鱼粉含量变化对试验鱼生长的影响。Ren等^[8]报道,在最适饲料蛋白质水平下以PBM和豆粕的混合物为替代原料时可将大口黑鲈饲料中鱼粉含量降低至16%;随后的研究表明以鸡血浆粉、发酵豆粕、γ射线辐照处理豆粕、γ射线辐照处理羽毛粉、去皮豆粕、虾水解物与植物蛋白质的混合物、酶处理大豆、PBM与豆粕的混合物等为替代原料时可将大口黑鲈饲料中鱼粉含量降低至41%~16%^{[9⇓⇓⇓⇓⇓⇓-16]}。Wang等^[17]报道,以PBM、PBM与棉籽浓缩蛋白的混合物或PBM与SPC和棉籽浓缩蛋白的混合物等为替代蛋白质源时可将大口黑鲈饲料中鱼粉含量降低至8%。Mclean等^[21]报道,利用PBM和SPC的混合物可完全替代大口黑鲈饲料中的鱼粉。本研究的试验1中,摄食F40、F8和F4的大口黑鲈WG和FBW无显著差异;与摄食F40的大口黑鲈相比,摄食F8的大口黑鲈WG和FBW分别下降1.5%和1.8%,摄食F4的大口黑鲈WG和FBW分别下降7.9%和9.4%。鉴于对鱼类生长差异进行统计学分析时同一处理的重复间(组内)变异程度会影响分析结果,王岩^[46]建议结合统计学分析结果和阈值方法判断鱼类生长差异是否显著。根据大口黑鲈养殖生产实际情况,设处理间相对生长差异的阈值为5%。摄食F4的大口黑鲈较摄食F40的大口黑鲈WG和FBW相对下降幅度超过7%,表明以PBM为替代蛋白质源时不能将大口黑鲈饲料中鱼粉含量降低至4%。这一结果与Wang等^[17]的结论一致,但不支持Mclean等^[21]关于大口黑鲈饲料中鱼粉可被完全替代的结论。由于PBM替代肉食性鱼类饲料中鱼粉的效果优于SPC^[17],初步认为利用SPC作为替代蛋白质源时大口黑鲈饲料中鱼粉最低含量不能低于8%。

本研究的试验1中,随饲料鱼粉替代水平升高,大口黑鲈的FI和FCR呈增加趋势,而NRE和ERE呈下降趋势(FI和ERE的变化趋势在统计学分析中不显著),表明利用PBM替代鱼粉会导致饲料利用效率降低,但不会使饲料适口性下降。FCR和NRE下降的原因可能与PBM的蛋白质消化率与鱼粉相比较低有关。与摄食F40的大口黑鲈相比,摄食F8的大口黑鲈FI、FCR、NRE和ERE无显著差异。早期研究表明,通过添加PBM将大口黑鲈和花鲈(Lateolabrax japonicus)饲料中鱼粉含量降低至8%不会导致NRE明显下降^[17,44]。本研究与此结论一致,摄食F4的大口黑鲈FCR显著高于摄食F40的大口黑鲈,前者NRE显著低于后者,表明将饲料中鱼粉含量降低至4%时不仅导致大口黑鲈生长缓慢,还会导致饲料利用效率明显下降。

必需氨基酸组成不平衡被认为是影响饲料营养和鱼粉替代水平的重要因素^[44]。近年来,动物饲料中功能性氨基酸对动物营养和生理的调控作用受到重视^[24,26]。本研究中,摄食F40、F8和F4的大口黑鲈ARE与NRE接近;蛋氨酸、甘氨酸和丙氨酸的贮积效率在单个氨基酸中相对较高;摄食F40的大口黑鲈甘氨酸和脯氨酸的贮积效率显著低于摄食F8和F4的大口黑鲈,其蛋氨酸的贮积效率也略低于后者(统计学差异不显著)。Wang等^[44]报道,花鲈的ARE与NRE接近,且赖氨酸、甘氨酸和丙氨酸的贮积效率与其他单个氨基酸的贮积效率相比较高(蛋氨酸的贮积效率因在试验饲料中补充了晶体蛋氨酸而下降),本研究结果与此结论一致。尽管动物体内可合成生长和代谢所需的氨基酸,氨基酸的贮积效率在一定程度上仍然可反映出饲料氨基酸供给是否充分^[44]。结合前人研究结果^[44]和本研究结果,初步认为进行低鱼粉饲料配方设计时应注意蛋氨酸、赖氨酸、甘氨酸和丙氨酸缺乏的问题。然而,摄食F8和F4的大口黑鲈之间各氨基酸的贮积效率无显著差异,说明蛋氨酸、甘氨酸和丙氨酸缺乏可能不是导致本研究中摄食F4的大口黑鲈生长减慢的主要原因。

3.2 降低饲料中动物蛋白质的比例对大口黑鲈生长和饲料利用效率的影响

Wang等^[47]报道,增加基础饲料中PBM含量可提高SPC对卵形鲳鲹(Trachinotus ovatus)饲料鱼粉的替代水平,表明饲料中动物蛋白质所占的比例可能是影响鱼粉替代水平的一个重要因素。本研究的试验2中,通过利用SPC、棉籽粕和DPS的混合物替代F8和F4中的PBM将饲料中动物蛋白质的比例从63%(F8和F4)分别降低到51%(F8D)和49%(F4D)。在相同饲料鱼粉含量下,摄食F8和F4的大口黑鲈WG和FBW分别大于摄食F8D和F4D的大口黑鲈(F8与F8D相比,F4与F4D相比),表明饲料中动物蛋白质所占比例是影响大口黑鲈生长的重要因素。

DPS中含有谷胱甘肽、精胺、精氨酸、腐胺、胍、肌酸、肌酸酐、肌肽、丝氨酸等多种成分^[25],其中小肽被认为可促进家畜和家禽的生长^[31]。研究表明,添加3% DPS替代鲤鱼(Cyprinus carpio)饲料中的鱼粉或添加5.6% DPS替代罗非鱼(Oreochromis niloticus)饲料中等量的鱼粉不会导致试验鱼生长速度发生明显变化^[33-34];添加5%的DPS替代黄鳝(Monopterus albus)饲料中的鱼粉可加快试验鱼的生长,而将DPS添加量降低至2.5%时导致试验鱼生长减慢^[35];在大黄鱼低鱼粉饲料中添加0.5% DPS可促进试验鱼的生长^[37]。在本研究的试验2中,摄食F8D和F4D(含0.5%的DPS)的大口黑鲈生长速度慢于摄食F8和F4的大口黑鲈,表明添加DPS难以弥补饲料中动物蛋白质比例下降对大口黑鲈生长所产生的负面影响。

3.3 添加水杨酸盐对提高大口黑鲈饲料鱼粉替代水平的作用

水杨酸盐具有清除机体内自由基、抗氧化的功能^[38],但尚未见其在水产饲料中应用的报道。本研究中,在F8D和F4D中添加0.005%水杨酸盐导致NRE增加,但未对大口黑鲈的WG、FBW、FI、FCR、PRE和ERE产生显著影响。与摄食F40的大口黑鲈相比,摄食F8D、F4D、F8DS和F4DS的大口黑鲈WG分别下降12.1%、15.7%、2.5%和5.4%,FBW分别下降9.9%、13.0%、2.0%和4.6%。摄食F4DS的大口黑鲈的生长下降幅度小于摄食F4的大口黑鲈(WG下降幅度:4.6% vs 7.9%;FBW下降幅度:5.4% vs 9.4%),这意味着添加水杨酸盐可改善摄食低鱼粉饲料的大口黑鲈的生长。根据本研究结果,推测通过添加水杨酸盐等功能性添加剂和改进配方的营养平衡有可能将大口黑鲈饲料鱼粉含量降低至4%,这一推测有待试验验证。

3.4 降低饲料鱼粉含量对大口黑鲈形态和体组成的影响

替代饲料中鱼粉可对鱼的CF、HSI、VSI和体组成产生一定的影响,但不同研究之间所得结果^[8,17,44]并不一致。例如,利用PBM与豆粕的混合物替代饲料中鱼粉导致大口黑鲈VSI显著降低,但未引起HSI发生明显变化^[8];利用PBM替代饲料中鱼粉导致大口黑鲈鱼体脂肪含量下降,但未使CF和HSI发生明显改变^[17];利用PBM替代饲料中鱼粉使花鲈CF和VSI显著增加^[44];利用PBM替代饲料中鱼粉未导致卵形鲳鲹CF、HSI、VSI以及鱼体粗蛋白质和粗脂肪含量产生显著影响^[48]。本研究中,与摄食F40的大口黑鲈相比,摄食F8、F4、F8D和F8DS的大口黑鲈CF、HSI和VSI无显著差异;摄食F4D的大口黑鲈HSI和VSI显著下降;摄食F4DS的大口黑鲈HSI和VSI显著增加;摄食F8、F4、F4D和F4DS的大口黑鲈鱼体粗蛋白质含量显著下降。这些结果表明,将饲料鱼粉含量从40%降低至8%时往往不会对大口黑鲈的CF、HSI和VSI产生显著影响,但当饲料鱼粉含量进一步降低至4%时则会导致鱼体形态和组成发生变化。有关大口黑鲈在饲料最低鱼粉含量下的鱼体形态和组成变化趋势还需要进一步研究。

3.5 降低饲料鱼粉含量对大口黑鲈养殖的自然资源与环境的影响

鱼类养殖对自然资源和环境的影响是决定养殖产业能否可持续发展的重要因素。对自然资源和环境的影响可分别用单位养殖产量的自然资源(土地、水、饲料蛋白质源等)消耗量和养殖废物排放量来定量评价。降低饲料鱼粉含量可降低FIFO。将大口黑鲈、花鲈和日本黄姑鱼(Nibea japonica)饲料鱼粉含量分别被降低至8%、8%和24%时,FIFO可分别降低至0.25~0.27^[17]、0.29^[44]和0.92 g鱼粉/g鱼产量^[49]。本研究中,投喂大口黑鲈F8、F8D和F8DS时,FIFO分别为0.30、0.31和0.26 g鱼粉/g鱼产量,这与Wang等^[17]的研究结果基本一致。替代饲料中鱼粉对养殖废物排放量的影响因鱼类种类、基础饲料组成、替代原料种类和饲料鱼粉替代水平不同而异。例如,利用PBM替代饲料中鱼粉增加了大口黑鲈养殖的PW,但对NW无显著影响^[17];相反,利用PBM替代饲料中鱼粉显著增加了卵形鲳鲹养殖的NW^[48]。本研究中,与投喂F40的大口黑鲈相比,投喂F8、F4、F8DS和F4DS的大口黑鲈NW、PW未发生显著变化,投喂F8D和F4D的大口黑鲈NW显著增加。这些结果表明,将饲料鱼粉含量降低至8%时不会加剧大口黑鲈养殖的氮、磷污染,而将饲料鱼粉含量降低至4%时有可能会使大口黑鲈养殖的氮污染加剧。

4 结论

以PBM为单一替代蛋白质源时,将饲料鱼粉含量降低至8%对大口黑鲈的生长、饲料利用效率、体组成和废物排放量无显著影响,将饲料鱼粉含量降低至4%则会导致大口黑鲈生长缓慢,饲料利用效率下降,NW增加。在低鱼粉饲料中添加0.005%水杨酸盐可改善大口黑鲈的生长和NRE,降低NW。根据本研究结果,在大口黑鲈养殖生产中应用无鱼粉饲料仍然面临挑战。

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