蟋蟀粉替代鱼粉对黄颡鱼幼鱼生长性能、肌肉氨基酸组成和血清生化指标的影响

引用本文

刘苑艺, Paul B. Brown, 裴卓, 刘慧, 樊可, 黄勇. 蟋蟀粉替代鱼粉对黄颡鱼幼鱼生长性能、肌肉氨基酸组成和血清生化指标的影响[J]. 渔业科学进展, 2023, 44(5): 69-79. DOI: 10.19663/j.issn2095-9869.20230213001.

LIU Yuanyi, Paul B. BROWN, PEI Zhuo, LIU Hui, FAN Ke, HUANG Yong. Effects of the Replacement of Fish Meal with Cricket Meal on Growth, Muscle Amino Acid Composition, and Serum Biochemical Indices of Juvenile Yellow Catfish (Pelteobagrus fulvidraco)[J]. Progress in Fishery Sciences, 2023, 44(5): 69-79. DOI: 10.19663/j.issn2095-9869.20230213001.

基金项目

河南省高等学校重点科研项目(23A240003)资助

作者简介

刘苑艺，E-mail: liuyuanyi0504@163.com

通讯作者

黄勇，副教授，E-mail: huangyong1979111@126.com

文章历史

收稿日期：2023-02-13
收修改稿日期：2023-03-31

Contents Abstract Full text Figures/Tables PDF

蟋蟀粉替代鱼粉对黄颡鱼幼鱼生长性能、肌肉氨基酸组成和血清生化指标的影响

刘苑艺 ¹, Paul B. Brown ², 裴卓 ¹, 刘慧 ¹, 樊可 ¹, 黄勇 ¹

1. 河南科技大学动物科技学院河南洛阳 471023;
2. 普渡大学森林与自然资源系美国西拉法叶市 47907

收稿日期：2023-02-13；收修改稿日期：2023-03-31

基金项目：河南省高等学校重点科研项目(23A240003)资助

作者简介：刘苑艺，E-mail: liuyuanyi0504@163.com.

通讯作者：黄勇，副教授，E-mail: huangyong1979111@126.com.

摘要：为研究蟋蟀(Gryllus bimaculatus)粉替代鱼粉对黄颡鱼(Pelteobagrus fulvidraco)幼鱼生长性能、肌肉成分和血清生化指标的影响，以黄颡鱼幼鱼[(2.0±0.13) g]为研究对象，分别使用蟋蟀粉替代0 (对照组)、15%、30%、45%和60%的鱼粉配制成5组等氮等能的实验饲料，分别记为T0、T15、T30、T45和T60组。实验幼鱼在室内循环水系统进行为期10周的养殖实验。结果显示，随着蟋蟀粉替代量的增加，黄颡鱼幼鱼生长呈先增加后降低的趋势；T30组的终末均重(FBW)、增重率(WGR)和特定生长率(SGR)最高，且均显著高于T0组；T30组的饲料系数(FCR)显著低于T0和T15组；T30的肝体比(HSI)显著高于T0和T15，而与T45和T60组无显著差异。与T0组相比，各替代组的脏体比(VSI)、摄食量(FI)、肥满度(CF)和成活率(SR)均无显著差异。黄颡鱼幼鱼肌肉必需氨基酸(EAA)含量中，T60组肌肉中精氨酸(Arg)和缬氨酸(Val)含量显著高于T0组；且蟋蟀粉替代不同比例的鱼粉对黄颡鱼幼鱼肌肉中的总呈味氨基酸(TFAA)含量均未产生显著影响。T30、T45和T60组血清中的葡萄糖(GLU)含量显著高于T0组；相反，其总胆固醇(TCHO)含量显著低于T0组。在本研究条件下，蟋蟀粉替代不同比例的鱼粉不影响黄颡鱼幼鱼的生长性能和肌肉氨基酸含量，且能增加血清GLU和降低TCHO含量，以30%的蟋蟀粉替代鱼粉比例能生长最佳，且优于对照组。本研究对揭示蟋蟀粉在黄颡鱼饲料中替代鱼粉应用的可行性、为今后昆虫蛋白源在水产饲料中的开发提供科学依据。

关键词：黄颡鱼蟋蟀粉生长性能肌肉成分血清生化指标

Effects of the Replacement of Fish Meal with Cricket Meal on Growth, Muscle Amino Acid Composition, and Serum Biochemical Indices of Juvenile Yellow Catfish (Pelteobagrus fulvidraco)

LIU Yuanyi ¹, Paul B. BROWN ², PEI Zhuo ¹, LIU Hui ¹, FAN Ke ¹, HUANG Yong ¹

1. College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471023, China;
2. Department of Forestry and Natural Resources, Purdue University, West Lafayette 47907, USA

Corresponding author: HUANG Yong, E-mail: huangyong1979111@126.com.

Abstract: In recent years, with the increase of varieties and the expansion of scale in aquaculture, as well as the rapid development of the intensive and industrial aquaculture industry, the demand for fish meal has increased significantly. On the other hand, due to global warming and environmental pollution, marine resources have reduced and the production of high-quality fish meal is gradually decreasing. The soaring prices of fish meal increase the feed cost in the process of aquaculture, severely decrease the economic benefits of aquaculture farmers, greatly limit the use of fish meal in aquatic feed, and hinder the sustainable development of the aquaculture industry. Therefore, it has become an important research subject in the aquatic feed industry to find new fish meal substitutes and reasonably reduce the amount of fish meal in feed. Crickets have high nutrient concentrations (55%–73% crude protein, high unsaturated fatty acid levels, and sufficient essential amino acid (EAA) profiles). For cricket meal, as one of the new high-quality insect protein sources, the crude protein concentration is comparable to that of fish meal. Recent studies have shown that cricket meal can replace part of fish meal, and have achieved good results in Clarias gariepinus, Micropterus salmoides, and other aquatic animals. However, the application of cricket meal as a substitute for fish meal in the diets of yellow catfish has not been reported. In the present study, we investigated the effects on the growth performance, muscle composition, and serum biochemical indexes of yellow catfish by replacing different proportions of fish meal in the diets with cricket meal. The aim was to explore the feasibility of replacing fish meal in the diets of yellow catfish, and to provide a scientific reference for the future development and application of insect protein sources in aquatic feed. The cricket meal used in this study was a brown powder containing dry matter crude protein content of 63.40%, crude fat content of 15.50%, and crude ash content of 7.36%. Healthy juvenile yellow catfish with an average body weight of (2.0±0.13) g were randomly divided into five groups with three replicates and 30 fish per replicate. Five isonitrogenous and isoenergetic experimental diets were formulated by replacing 0%, 15%, 30%, 45%, and 60% of fish meal protein with cricket meal, named T0, T15, T30, T45, and T60 groups, respectively. The experimental fish were reared in an indoor recirculating aquaculture system for 10 weeks. By measuring growth performance, muscle amino acid content, and serum biochemical parameters, the appropriate replacement level of cricket meal in yellow catfish diets was investigated.The results showed that with increasing cricket meal content, the final body weight (FBW), weight gain rate (WGR), and specific growth rate (SGR) of juvenile yellow catfish increased first and then decreased. The growth performance of FBW, WGR, and SGR in the T30 group was the highest and significantly higher than that of FBW, WGR, and SGR in the T0 group (P < 0.05), whereas the feed conversion rate (FCR) was significantly lower than that of T0 and T15 groups (P<0.05). The hepatosomatic index in the T30 group was higher than that in the T0 and T15 groups (P<0.05), and there was no difference between the T45 and T60 groups. There were no significant differences in the viscerosomatic index, feed intake, FCR, and survival rate among all groups (P>0.05). The EAA contents of the muscle arginine and valine in the T60 group were significantly higher than those in the T0 group (P<0.05). There were no significant differences in the contents of total flavor amino acid in muscle among all groups with different proportions of replaced cricket meal (P>0.05). Compared with the T0 group, the content of glucose (GLU) in the serum of the T30, T45, and T60 groups significantly increased (P<0.05), whereas the content of total cholesterol (TCHO) was significantly decreased (P<0.05). The activities of the serum superoxide dismutase and catalase in the T30 and T60 groups were significantly higher than those of the T0 group (P<0.05). In conclusion, under our experimental conditions, the growth performance and muscle amino acid composition of juvenile yellow catfish were not affected by replacing fish meal with cricket meal, and serum biochemical parameters and TCHO contents were increased. The optimal growth rate was achieved by replacing fish meal with 30% cricket meal. The results of indicate that cricket meal is an excellent substitute for fish meal and provides a theoretical reference for the application of cricket meal as a partial substitute for fish meal in aquatic animal diets.

Key words: Pelteobagrus fulvidraco Cricket meal Growth performance Muscle composition Serum biochemical indices

鱼粉因其营养丰富、氨基酸配比均衡、适口性好、维生素和矿物质丰富和易于被消化吸收等优点，长期以来一直是水产饲料中的重要蛋白源。近年来，随着养殖品种的增加和养殖规模扩大以及工厂化水产养殖业的快速发展，对鱼粉的需求量增加；另一方面，由于全球变暖和环境污染加剧，导致海洋资源减少，鱼粉的产量波动幅度较大，使鱼粉价格不断飙升，不但增加了养殖户的饲料成本，而且极大地限制了鱼粉在水产饲料中的使用量，直接影响了水产养殖业的可持续发展(钱妤等, 2020; 周歧存等, 2005; Henry et al, 2015; 王成强等, 2021)。因此，寻求廉价、安全、来源广泛且环境友好型的鱼粉替代蛋白源已成为当前水产养殖业亟待解决的问题(Hardy, 2010; Tacon et al, 2008)。

蟋蟀(Gryllulus)属无脊椎动物(Invertebrate)、昆虫纲(Insecta)、直翅目(Orthoptera)、蟋蟀科(Gryllidae)。世界上已定名的蟋蟀约有1 400种以上，我国有30种以上且已被定名，在全国都有分布(马俊等, 2019)。蟋蟀具有食性杂，环境适应性强、繁殖快且容易饲养等特点。在种类繁多的蟋蟀中，双斑蟋蟀(Gryllus bimaculatus)是在亚洲、非洲和欧洲的热带和亚热带地区发现的数量最多的蟋蟀物种之一。目前，一些国家已经进行了蟋蟀的大规模人工养殖，并形成了一定的产业链(Magara et al, 2020; Verneau et al, 2021)。如泰国从1997年开始人工饲养蟋蟀，在2010年全国的年产量已达7 500 t (Halloran et al, 2016)。研究表明，双斑蟋蟀水分含量为71.0%，干重时的粗蛋白、粗脂肪、粗纤维和灰分含量分别为58.60%、28.90%、7.23%和4.93%，远高出其他昆虫粗蛋白的含量，且含有维生素B₁₂、铁元素、丰富的钙、高不饱和脂肪酸等(何钊等, 2021; Moreki et al, 2012)，被认为是新型优质昆虫蛋白源之一。Taufek等(2016)研究表明，用蟋蟀粉替代饲料中(含35%粗蛋白) 75%和100%的鱼粉能改善非洲鲶(Clarias gariepinus)幼鱼的生长性能，表明蟋蟀粉能作为鱼类养殖中可替代鱼粉的蛋白源。Kilburn等(2020)研究表明，蟋蟀粉亦能作为一种高蛋白源添加在宠物饲料中，而且24%的添加量对成年宠物狗的生长无不利影响。

黄颡鱼(Pelteobagrus fulvidraco)俗称“黄骨鱼、黄嘎牙和黄辣丁”等，是我国江河水域中分布较广的一种广温性淡水经济鱼类。因其肉质细嫩、味道鲜美、无肌间刺和营养价值高等特点，深受消费者的喜爱，现已成为主要的名特优鱼类品种之一(王秀娟等, 2019)。黄颡鱼是一种杂食偏肉食性鱼类，且幼鱼饲料对鱼粉需求量较大，所占比例超过35% (Ye et al, 2009)。目前，蟋蟀粉替代鱼粉在黄颡鱼饲料中的应用研究未见报道。本研究以黄颡鱼幼鱼为研究对象，通过蟋蟀粉替代饲料中不同比例的鱼粉，研究对其生长性能、肌肉成分以及血清生化指标的影响，旨在探讨蟋蟀粉在黄颡鱼饲料中替代鱼粉的可行性，为今后昆虫蛋白源在水产饲料中的开发应用提供科学依据。

1 材料与方法 1.1 实验鱼

实验用黄颡鱼为河南洛阳吉利区某渔场的同一批鱼苗，体质量约为(2.0±0.13) g。选择体质健壮的实验幼鱼450尾，用杀菌盐水(盐度为3)消毒后，放入暂养池中养殖。

1.2 实验蟋蟀粉和饲料

实验用双斑蟋蟀购自安徽亳州中药材批发市场，烘干并粉碎后于低温保存备用。经测定，蟋蟀粉的粗蛋白、粗脂肪和粗灰分含量分别为63.40%、15.50%和7.36%；以鱼粉、血粉和豆粕为主要蛋白源，鱼油和豆油混合油为主要脂肪源，高筋面粉为糖源配制基础饲料(含40.00%的鱼粉)。在此基础上，用蟋蟀粉分别代替基础饲料中0、15%、30%、45%和60%的鱼粉，配制成5组等氮(粗蛋白含量为44.0%)和等能(19.00 MJ/kg)的实验饲料，命名依次为T0组(0替代)、T15组(15%替代)、T30组(30%替代)、T45组(45%替代)和T60组(60%替代)。实验饲料配方及营养组成见表 1，鱼粉、蟋蟀粉和各组饲料中氨基酸组成见表 2。饲料原料粉碎后80目过筛，按比例称量鱼粉和蟋蟀粉，混匀后加入一定比例的蒸馏水，使用QRLS-400型电动绞肉机制成粒径为2.5 mm颗粒饲料，烘干并在阴凉处冷却后再装入白色透明密封袋，4 ℃冰箱中存储备用。

表 1 实验饲料配方及营养组成(%风干基础) Tab.1 Formulation and approximate composition of experimental diets (% air-dry basis)

项目 Items	实验组 Groups
项目 Items	T0	T15	T30	T45	T60
原料 Ingredients
鱼粉 Fish meal	40.00	34.00	28.00	22.00	16.00
蟋蟀粉 Cricket meal	0.00	6.00	12.00	18.00	24.00
豆粕 Soybean meal	10.00	10.00	10.00	10.00	10.00
玉米蛋白粉 Corn gluten meal	10.00	10.00	10.00	10.00	10.00
血粉 Blood meal	4.50	4.50	4.50	4.50	4.50
小麦粉 Wheat flour	25.90	25.90	25.90	25.90	25.90
混合油 Mixed oils¹	5.00	5.00	5.00	5.00	5.00
乌贼膏 Squid ointment	1.00	1.00	1.00	1.00	1.00
预混料 Premix²	1.00	1.00	1.00	1.00	1.00
氯化胆碱 Choline chloride	0.50	0.50	0.50	0.50	0.50
磷酸二氢钙 Ca(H₂PO4)₂	1.10	1.10	1.10	1.00	1.00
羧甲基纤维素钠 Sodium carboxymethyl cellulose	1.00	1.00	1.00	1.00	1.00
总计 Total	100	100	100	100	100
营养水平 Nutrient levels³
水分 Moisture	9.45	9.50	9.51	9.64	9.75
粗蛋白 Crude protein	44.28	44.29	44.17	43.09	44.03
粗脂肪 Crude lipid	12.31	12.25	12.21	12.18	12.15
总能 Gross energy (MJ/kg)	19.43	19.39	19.28	19.17	19.15
注：1. 混合油为1∶1的豆油和鱼油；2. 预混料为每千克饲料提供的微量元素和维生素：钾30 mg，钙300 mg，镁 10 mg，锌3 mg，铁22 mg，铜1.7 mg，碘0.29 mg，硒0.02 mg，维生素A 8 000 IU，维生素C 230 mg，维生素D 800 IU，维生素E 90 mg，维生素K 8 mg，维生素B₁ 10 mg，尼克酸100 mg，泛酸钙50 mg，维生素B₆ 15 mg，维生素B₁₂ 10 mg，肌醇150 mg；3. 营养成分为实测值。 Note: 1. Mixed oils: 1:1 mixture of soybean oil and fish oil; 2. The premix provides mineral and vitamin for a kilogram of diets: K 30 mg, Ca 300 mg, Mg 10 mg, Zn 3 mg, Fe 22 mg, Cu 1.7 mg, I 0.29 mg, Se 0.02 mg, V_A 8 000 IU, V_C 230 mg, V_D 800 IU, V_E 90 mg, V_K 8 mg, V_B1 10 mg, nicotinic acid 100 mg, pantothenate acid 50 mg, V_B6 15 mg, V_B12 10 mg, inositol 150 mg; 3. The feed nutrition level were measured values.

表 1 实验饲料配方及营养组成(%风干基础) Tab.1 Formulation and approximate composition of experimental diets (% air-dry basis)

表 2 蟋蟀粉、鱼粉和实验饲料的氨基酸组成(%风干基础) Tab.2 Amino acid composition of fish meal, cricket meal and the experimental diets (% air-dry basis)

1.3 实验设计与实验鱼饲养管理

养殖实验在河南科技大学动物科技学院水生生物实验室循环水养殖系统中进行。养殖系统由15个120 cm×35 cm×40 cm的长方体玻璃纤维鱼缸组成，养殖实际水容量为150~170 L，养殖用水为已曝气的自来水。黄颡鱼幼鱼用基础饲料(粤海饲料)驯养2周后再分组，正式实验开始前，禁食24 h。挑选状态良好，体表无伤的黄颡鱼幼鱼450尾，随机分为5个处理组，每个处理组3个重复，每个重复30尾鱼，测定初始体质量与体长；分别投喂对照组T0和蟋蟀粉替代鱼粉组T15、T30、T45和T60饲料，投食观察直至幼鱼饱食，每天投喂2次(08:00和17:30)。养殖周期为10周，为保持水质良好，每天换水1次，换水量为总体积的1/3。实验期间保持水温为25.0~30.0 ℃，pH为7.8~8.0，溶解氧>5.0 mg/L，氨氮<0.05 mg/L，亚硝酸盐<0.1 mg/L。

1.4 样品采集

养殖实验结束后，禁食24 h，用丁香酚(1∶10 000)麻醉后，称量鱼体总重，测量体长并计数，计算生长性能。每个缸随机抽取6尾幼鱼，采用1 mL无菌注射器在尾静脉取血，随后，在冰上解剖分离内脏团和肝脏并称重；抽取的血液静置3 h后，离心(4 000 r/min) 15 min，取上清液，于−80 ℃超低温冰箱中保存，用于后续血清生化指标的测定；取幼鱼同侧背肌，于−80 ℃超低温冰箱中保存，用于后续氨基酸组成测定。

1.5 常规成分和氨基酸组成测定

蟋蟀粉和饲料的水分、粗灰分、粗蛋白质和粗脂肪分别采用恒温常压干燥法(GT/T 6435-2014)、550 ℃马弗炉灼烧法(GB/T 6438-2007)、凯氏定氮法(GB/T 6432- 2018)和索氏抽提法(GB/T 6433-2006)进行测定。蟋蟀粉、鱼粉、饲料和肌肉氨基酸组成用盐酸水解法前处理后用氨基酸自动分析仪(日立L-8900型, 日本)测定。

1.6 生长性能指标与计算公式

成活率(survival rate, SR, %)=N_t/N₀×100%；

增重率(weight gain, WG, %)=(W_t−W₀)/W₀×100%；

特定生长率(specific growth rate, SGR, %/d)=(lnW_t−ln W₀)/t×100%；

饲料系数(feed conversion ratio, FCR)=F/(W_t−W₀)；

肝体比(hepatopancreas somatic index, HSI, %)=W_h/W×100%；

脏体比(viscerosomatic index, VSI, %)=W_v/W×100%；肥满度(condition factor, CF)=W_t/L_t³×100；

摄食量(feed intake, FI, g/尾)=投饲总量/[(N₀+N_t)/2］。

式中，N₀为实验初始各组鱼的尾数；N_t为实验终末各组鱼的尾数；W₀为实验初始各组鱼的体质量(g)；W_t为实验终末各组鱼的体质量(g)；t为养殖实验天数(d)；F为饲料摄入量(g)；L_t为实验结束时鱼的体长(cm)；W为鱼体质量(g)；W_h为鱼体肝脏质量(g)；W_v为鱼体内脏团质量(g)。

1.7 血清生化指标测定

血清生化指标采用日立7100全自动生化分析仪测定。测定指标有谷丙转氨酶(alanine aminotransferase, ALT)、谷草转氨酶(aspartate transaminase, AST)、总蛋白(total protein, TP)、尿氮(urea nitrogen, UN)、葡萄糖(glucose, GLU)、甘油三酯(triglyceride, TG)、总胆固醇(total cholesterol, TCHO)和丙二醛(malondialdehyde, MDA)含量(由河南科技大学第一附属医院新区医院检验中心测定完成)。

1.8 数据统计分析

实验数据首先使用Excel 2016软件进行整理，然后采用SPSS 20.0软件进行单因素方差分析(one-way ANOVA)，若有组间差异，再采用Duncan´s检验法进行多重比较，数据用平均值±标准差(Mean±SD)表示，P<0.05为差异显著。

2 结果与分析 2.1 蟋蟀粉替代鱼粉对黄颡鱼幼鱼生长性能的影响

由表 3可知，与T0相比较，T15~T60组的脏体比、摄食量、肥满度和成活率在黄颡鱼幼鱼间均无显著差异(P>0.05)。T30组的终末体质量、增重率和特定生长率与T0组相比增加显著(P<0.05)，与之相反，饲料系数显著降低(P<0.05)；T15、T45和T60组的终末体质量、增重率和特定生长率与T0组差异不显著(P>0.05)。与T0和T15组相比，T30组的肝体比显著增加(P<0.05)，而与T45和T60组差异不显著，但呈降低趋势(P>0.05)。

表 3 蟋蟀粉替代鱼粉对黄颡鱼幼鱼生长性能的影响 Tab.3 Effects of fish meal replacement by cricket meal on the growth performance of juvenile yellow catfish

2.2 蟋蟀粉替代鱼粉对黄颡鱼幼鱼肌肉氨基酸组成的影响

蟋蟀粉的营养成分见表 4。由表 4可知，必需氨基酸中的赖氨酸和蛋氨酸含量略低于鱼粉；而精氨酸含量较鱼粉高。对黄颡鱼幼鱼肌肉进行氨基酸成分测定(表 4)，共检测出9种必需氨基酸和8种非必需氨基酸。蟋蟀粉替代不同比例的鱼粉对黄颡鱼幼鱼肌肉中的必需氨基酸：组氨酸、苏氨酸、蛋氨酸、苯丙氨酸、异亮氨酸、亮氨酸和赖氨酸以及必需氨基酸的总量和总非必需氨基酸的含量均无显著影响(P>0.05)；而T60组中的精氨酸和缬氨酸含量显著高于T0和T15组(P<0.05)。在非必需氨基酸中，各实验组肌肉中谷氨酸含量与T0组相比差异不显著(P>0.05)，除T60组含量显著低于T0组外(P<0.05)，其他非必需氨基酸含量在各组间差异不显著(P>0.05)。此外，蟋蟀粉替代不同比例鱼粉对黄颡鱼幼鱼肌肉中的天冬氨酸、甘氨酸和丙氨酸以及总氨基酸的含量也均未产生显著影响(P>0.05)。

表 4 蟋蟀粉替代鱼粉对黄颡鱼幼鱼的肌肉氨基酸组成的影响(%风干基础) Tab.4 Effects of fish meal replacement by cricket meal on muscle amino acid composition in juvenile yellow catfish (% air-dry basis)

项目 Items	实验组 Groups
项目 Items	T0	T15	T30	T45	T60
必需氨基酸 EAA
组氨酸 His	1.59±0.10	1.55±0.13	1.51±0.14	1.50±0.17	1.48±0.19
苏氨酸 Thr	1.47±0.44	1.45±0.06	1.43±0.04	1.41±0.01	1.39±0.36
精氨酸 Arg	1.45±0.65^a	1.48±0.03^a	1.62±0.02^ab	1.61±0.01^ab	1.79±0.22^b
缬氨酸 Val	3.33±0.88^a	3.39±0.03^a	3.40±0.01^a	3.47±0.03^ab	3.60±0.35^b
蛋氨酸 Met	1.55±0.01	1.53±0.01	1.52±0.03	1.51±0.02	1.46±0.06
苯丙氨酸 Phe	0.86±0.44	0.88±0.07	0.86±0.08^a	0.84±0.11	0.80±0.12
异亮氨酸 Ile	1.09±0.02	1.09±0.04	1.10±0.03	1.11±0.05	1.06±0.20
亮氨酸 Leu	4.68±1.14	4.70±0.04	4.69±0.03	4.71±0.02	4.58±0.32
赖氨酸 Lys	2.07±0.05	2.01±0.05	2.01±0.03	1.93±0.07	1.92±0.04
非必需氨基酸 NEAA
天冬氨酸 Asp^★	4.75±0.90	4.77±0.05	4.73±0.05	4.76±0.02	4.77±0.39
谷氨酸 Glu^★	7.12±1.61^b	7.10±0.07^b	7.01±0.07^ab	6.83±0.04^ab	6.55±0.47^a
丝氨酸 Ser	1.73±0.36	1.74±0.02	1.71±0.02	1.68±0.01	1.79±0.13
甘氨酸 Gly^★	2.79±0.03	2.74±0.02	2.71±0.03	2.70±0.03	2.69±0.05
丙氨酸 Ala^★	3.40±0.02	3.43±0.02	3.45±0.02	3.46±0.01	3.47±0.03
酪氨酸 Try	1.57±0.02	1.62±0.01	1.59±0.02	1.59±0.03	1.57±0.05
胱氨酸 Cys	0.12±0.02	0.12±0.03	0.15±0.01	0.15±0.01^b	0.14±0.01
脯氨酸 Pro	2.78±0.08	2.73±0.09	2.81±0.03	2.79±0.05	2.85±0.02
总呈味氨基酸 TFAA	18.06±0.17	18.14±0.10	17.90±0.19	17.75±0.18	17.48±0.25
总必需氨基酸 TEAA	18.10±0.21	18.08±0.08	18.16±0.06	18.09±0.07	18.08±0.04
总非必需氨基酸 TNEAA	24.26±0.11	24.25±0.13	24.16±0.16	23.96±0.14	23.83±0.15
注：★表示呈味氨基酸。 Note: ★denotes flavor amino acid (FAA).

2.3 蟋蟀粉替代鱼粉对黄颡鱼幼鱼血清生化指标的影响

由表 5可知，与T0组相比，随着蟋蟀粉替代比例的升高，各替代组的谷丙转氨酶、谷草转氨酶、总蛋白、尿氮和甘油三酯含量无显著差异(P>0.05)。T30组的葡萄糖含量与T0组相比显著增加(P<0.05)，但与T15、T45和T60差异不显著(P>0.05)。此外，T30组的总胆固醇含量显著低于对照组T0(P<0.05)，但其含量在各替代组间均无显著差异(P>0.05)；丙二醛含量在所有组间无显著差异(P>0.05)。

表 5 蟋蟀粉替代鱼粉对黄颡鱼幼鱼血清生化指标的影响 Tab.5 Effects of fish meal replacement by cricket meal on the serum biochemical indices of juvenile yellow catfish

3 讨论 3.1 蟋蟀粉替代鱼粉对黄颡鱼幼鱼生长性能的影响

当前，其他蛋白源作为鱼粉的替代品在黄颡鱼或其他养殖鱼的应用研究已有相关报道，如黄粉、蝇蛆、家蚕、磷虾粉和黑水虻(Hermetia illucens)的替代(贲玲芝等, 2022; 陈晓瑛等, 2019; 吉红等, 2016; 饶远等, 2019; 文远红等, 2013; 谢凯等, 2022; Su et al, 2017)。然而，蟋蟀粉作为昆虫蛋白源替代鱼粉在黄颡鱼幼鱼中的研究鲜有报道。蟋蟀粉是一种营养价值较高的昆虫蛋白源，具备成为功能食品和饲料蛋白源的潜力(Barroso et al, 2014; Maiyo et al, 2022; Makkar et al, 2014)。增重率和特定生长率是养殖鱼类生产中重要的参考指标，在不影响生长性能的前提下，本研究养殖10周结束时，蟋蟀粉替代基础饲料中鱼粉的30%时，黄颡鱼幼鱼的终末体质量、增重率、特定生长率、饲料系数和肝体比均显著增加；而当替代比例为45%和60%时，终末体质量、增重率、特定生长率、饲料系数和肝体比与对照组相比有下降趋势，但均无显著影响。蟋蟀粉替代鱼粉后也并不影响幼鱼的成活率，表明实验黄颡鱼能很好的适应蟋蟀粉替代鱼粉的饲料。值得注意的是，随着蟋蟀粉替代比例的增加，黄颡鱼幼鱼的终末体质量、增重率和特定生长率均呈现先上升后下降趋势，这可能与蟋蟀粉中含有几丁质而影响饲料的消化吸收有关。研究结果与Taufek等(2016)的研究不一致。Taufek等(2016)研究发现，利用蟋蟀粉分别替代非洲鲶饲料中75%和100%的鱼粉，饲养7周后，75%和100%替代的特定生长率较对照组全鱼粉组差异显著(P<0.05)，且特定生长率随着蟋蟀粉替代量的增加呈显著上升趋势，而饲料系数无显著差异。产生这种结果的原因可能有3种：第一，研究所用的蟋蟀粗蛋白、粗脂肪、粗灰分以及粗纤维含量不同。如家蟋蟀(Acheta domesticus)的粗蛋白和粗纤维要低于双斑蟋蟀(Gryllus bimaculatus)；而粗脂肪和粗灰分则要高(Champika et al, 2022)；第二，不同种鱼类的营养代谢机理不同，因而对蟋蟀粉的耐受能力也不同，导致蟋蟀粉在不同鱼类饲料中添加的适宜比例不同；第三，蟋蟀粉中含有的几丁质以及未知的抗营养因子，替代后饲料的适口性可能发生改变，影响机体对饲料的消化吸收(Alegbeleye et al, 2012; Finke et al, 1987; Wang et al, 2005)。针对这一现象，具体机理有待后续深入研究。本实验室前期研究表明，在蟋蟀粉替代大口黑鲈(Micropterus salmoides)幼鱼饲料中鱼粉(基础饲料中鱼粉含量为60%)的比例≤45%时，对终末体质量、增重率、特定生长率和饲料系数并无显著影响(Wang et al, 2022)，该研究也表明蟋蟀粉替代鱼粉饲喂后能影响不同种类鱼的生长性能。肝体比随蟋蟀粉替代鱼粉比例的升高而逐渐升高，替代比例为30%组的肝体比显著高于对照组，推测这可能与蟋蟀脂肪含量较高有关。研究表明，饲料中脂肪含量的高低显著影响鱼类的肝体比(Lee et al, 2002; Rueda-Jasso et al, 2004)，具体研究仍需要进一步开展。

3.2 蟋蟀粉替代鱼粉对黄颡鱼幼鱼肌肉氨基酸组成的影响

动物肌肉品质是和其本身的营养成分含量与组成密切相关，氨基酸是蛋白质的基本组成单位，他们的组成、种类和所占比例共同决定了肌肉蛋白质的营养价值。Jahan等(2021)研究表明，动物组织氨基酸含量主要受饲料中粗蛋白和氨基酸含量的影响。本研究对黄颡鱼幼鱼肌肉氨基酸进行了测定，鉴定出9种必需氨基酸和8种非必需氨基酸。进一步研究发现，蟋蟀粉替代不同比例的鱼粉对黄颡鱼幼鱼肌肉中大部分必需氨基酸和非必需氨基酸以及总呈味氨基酸、总必需氨基酸和总非必需氨基酸含量无显著影响。就必需氨基酸而言，在蟋蟀粉替代60%鱼粉的时候，精氨酸和缬氨酸含量均显著高于对照组，结果表明，高比例的蟋蟀粉替代鱼粉能影响部分幼鱼肌肉的氨基酸的组成。值得注意的是，第一限制性氨基酸的蛋氨酸在蟋蟀粉中的含量较鱼粉低，是否会对机体的氨基酸平衡产生影响(韩星星等, 2020)，仍有待于深入研究。同样在非必需氨基酸中，替代比例≤45%时，谷氨酸含量无明显变化。分析产生差异的原因可能是与饲料中鱼粉和蟋蟀粉含量呈正相关。谷氨酸为鲜味氨基酸，本研究中鱼粉和蟋蟀粉中谷氨酸的含量都显著高于其他氨基酸。此外，推测还可能与黄颡鱼幼鱼个体的增长、对饲料消化吸收的利用能力有关。本研究中，各实验组间总呈味氨基酸的含量无显著差异，表明蟋蟀粉替代鱼粉对黄颡鱼幼鱼的肌肉总氨基酸组成未产生显著影响；而且饲料中蟋蟀粉高比例替代不影响黄颡鱼幼鱼生长性能的原因可能是蟋蟀粉中含有精氨酸、蛋氨酸和赖氨酸等必需氨基酸。研究表明，这些必需氨基酸缺乏，则会降低水产动物的生长性能(Rossi et al, 2012)，因为精氨酸、蛋氨酸和赖氨酸等参与众多机体代谢过程，亦是合成胆碱和胱氨酸的前体物质(Goff et al, 2004; Kasper et al, 2000)。当前有关昆虫粉在水产动物中的氨基酸吸收与代谢机制尚不清楚。

3.3 蟋蟀粉替代鱼粉对黄颡鱼幼鱼血清生化指标的影响

血清生化指标可直观地反映鱼类生理机能和营养代谢状态。本研究中，蟋蟀粉替代鱼粉后，与对照组相比，其他各组血清中的谷丙转氨酶、谷草转氨酶、总蛋白、尿氮和甘油三酯含量均无显著变化。血清中谷丙转氨酶和谷草转氨酶含量可间接反映肝脏损伤状况。本研究中，所有组的谷丙转氨酶和谷草转氨酶含量都无显著变化，但略呈降低趋势，表明蟋蟀粉替代鱼粉后对鱼体肝脏并无负面的应激作用或者产生组织损伤现象。血清总蛋白是在肝中合成，其含量是反映机体内蛋白质代谢和营养健康的重要指标(孙宏等, 2014)。本研究中，黄颡鱼幼鱼血液总蛋白浓度在各组间无显著变化，说明蟋蟀粉替代不同比例鱼粉对鱼体的蛋白质营养与代谢无不利影响。血清甘油三酯含量在各组间也无显著变化，说明蟋蟀粉替代部分鱼粉不影响黄颡鱼幼鱼机体的脂质代谢。在蟋蟀粉替代30%~60%鱼粉时，葡萄糖含量无显著变化，但显著高于对照组，在替代30%鱼粉时达最大值，推测蟋蟀粉适量比例的替代鱼粉可能会提高黄颡鱼幼鱼肝脏的糖原合成能力，进而增加血清中葡萄糖的含量。随着蟋蟀粉比例的增加，总胆固醇呈降低趋势，与对照组相比，替代30%、40%和60%鱼粉时，总胆固醇显著降低。这与Magalhães等(2017)在黑水虻虫粉替代鱼粉饲喂欧洲鲈(Dicentrarchus labrax)能降低血浆中胆固醇含量的研究结果相一致。血清中总胆固醇主要源于肝脏，其含量的升高说明动物机体肝脏细胞功能发生障碍或损伤，机体脂质代谢发生紊乱。推测可能是蟋蟀粉胆固醇含量低于鱼粉，造成替代饲料中的胆固醇含量降低，从而引起黄颡鱼幼鱼血清胆固醇水平降低(王淑雯等, 2015)。

4 结论

在本研究条件下，蟋蟀粉替代不同比例的鱼粉不影响黄颡鱼幼鱼的生长性能和肌肉氨基酸含量，且能增加血清中葡萄糖和降低总胆固醇含量，以30%的蟋蟀粉替代鱼粉比例黄颡鱼幼鱼生长最佳。

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