生物絮团养殖模式下益生菌添加对异育银鲫生长、消化酶活性及肠道组织结构的影响

前期研究表明,生物絮团技术(biofloc technology,BFT)适于异育银鲫(Carassius auratus gibelio)养殖。为进一步优化BFT养殖模式,本研究设置3个实验组:BFT模式下EM菌添加组(BB组)、枯草芽孢杆菌(Bacillus subtilis)添加组(BI组)和BFT对照组(B组),以均体重(1.60±0.50)g的异育银鲫为研究对象,探讨BFT模式下外源添加益生菌对养殖动物生长、消化酶活性及肠道组织结构的影响。结果表明:(1)益生菌添加组异育银鲫增重率和特定生长率显著高于对照组(P0.05),BB和BI组的增重率分别提高了216.70%和184.04%,特定生长率分别提高了141.18%和125.49%,BB和BI组间差异不显著(P0.05);(2)益生菌添加组(BB组和BI组)的消化酶(淀粉酶、脂肪酶和胃蛋白酶)活性均显著高于对照组(B组)(P0.05)。益生菌添加组间,BB组淀粉酶活性显著高于BI组(P0.05),脂肪酶和胃蛋白酶活性亦高于BI组,但差异不显著(P0.05);(3)益生菌添加组肠道肌层厚度和黏膜下层厚度显著高于对照组(B组)(P0.05),BB组异育银鲫肠道黏膜皱襞高度和皱襞间质宽度与BI和对照组相比,均无显著差异(P0.05)。研究表明,BFT养殖模式下外源添加益生菌可以更好地促进异育银鲫生长。     

有机养殖模式下中华绒螯蟹抗氧化防护和溶菌酶的活性

比较了有机和常规养殖模式下,体质量165~175g中华绒螯蟹鳃、肝胰脏和肌肉中溶菌酶活性、超氧化物歧化酶、过氧化氢酶和谷胱甘肽过氧化物酶活性以及丙二醛含量。结果显示,有机养殖模式下,中华绒螯蟹肝胰脏和肌肉中超氧化物歧化酶和过氧化氢酶活性显著高于常规养殖模式(P0.05),但鳃中超氧化物歧化酶和过氧化氢酶活性与常规养殖组差异不显著(P0.05);鳃、肝胰脏和肌肉中谷胱甘肽过氧化物酶活性均显著高于常规养殖模式(P0.05);鳃、肝胰脏和肌肉中丙二醛含量显著低于常规养殖模式(P0.05)。有机养殖模式下中华绒螯蟹肝胰脏中的溶菌酶活性显著高于常规养殖组(P0.05)。试验结果表明,有机养殖模式下中华绒螯蟹具有较高的抗氧化防护能力和抗菌能力。     

循环水及生物絮团系统对大规格罗非鱼种生长及水质的影响

为比较越冬期循环水系统(RAS)与生物絮团系统(BFT)两种模式下大规格罗非鱼(Oreochromis niloticus)鱼种的生长性能与养殖水质的差异。选择RAS组与BFT组,进行为期67 d的养殖,测定养殖过程中罗非鱼种的生长情况以及水质的变化情况。结果显示,RAS组与BFT组增重率和特定增长率分别为(870.69±33.25)%、(3.39±0.05)%/d和(659.47±62.84)%、(3.02±0.13)%/d,RAS组显著高于BFT组;在水质控制方面,RAS组氨氮和亚硝酸盐从养殖初期到实验结束均维持在较低水平,而BFT组在初期氨氮和亚硝酸盐有升高的趋势,峰值分别达到了(4.53±0.72)mg/L和(6.68±1.8)mg/L,分别在第3天和第6天下降到较低水平,硝酸盐两组均呈现不断积累的趋势。结果表明,RAS系统养殖罗非鱼生长速度要高于BFT系统,RAS系统在水质控制方面略优于BFT系统。     

生物絮团对团头鲂(Megalobrama amblycephala)生长、消化酶和免疫相关酶活性的影响

为了研究零换水条件下团头鲂(Megalobrama amblycephala)养殖水体生物絮团形成所需的适合的碳氮比(C/N),以及不同C/N形成的生物絮团对团头鲂生长、消化酶活性和非特异性免疫力的影响,本实验设计4个不同C/N实验组,包括投喂基础饲料(C/N=8∶1)的对照组,在基础饲料上添加葡萄糖的处理组,其中将处理组的C/N分别调整为12∶1(C/N12)、16∶1(C/N16)和20∶1(C/N20).结果显示,C/N16和C/N20处理组中团头鲂的增重率和特定生长率显著高于对照组(P＜0.05),而饲料系数显著低于对照组(P＜0.05);C/N16和C/N20处理组中团头鲂肠道的蛋白酶活性和淀粉酶活性显著高于对照组(P＜0.05);而各实验组中团头鲂肠道的脂肪酶活性没有显著性差异;C/N16和C/N20处理组中团头鲂肝脏超氧化物歧化酶、碱性磷酸酶和溶菌酶活性显著高于对照组(P＜0.05).研究表明,生物絮团技术应用于团头鲂养殖适宜的C/N应不低于16,该条件下形成的生物絮团能有效提高团头鲂生长、消化酶和免疫相关酶活性.     

生物絮团对团头鲂(Megalobrama amblycephala)生长、消化酶和免疫相关酶活性的影响

为了研究零换水条件下团头鲂(Megalobrama amblycephala)养殖水体生物絮团形成所需的适合的碳氮比(C/N),以及不同C/N形成的生物絮团对团头鲂生长、消化酶活性和非特异性免疫力的影响,本实验设计4个不同C/N实验组,包括投喂基础饲料(C/N=8∶1)的对照组,在基础饲料上添加葡萄糖的处理组,其中将处理组的C/N分别调整为12∶1(C/N12)、16∶1(C/N16)和20∶1(C/N20)。结果显示,C/N16和C/N20处理组中团头鲂的增重率和特定生长率显著高于对照组(P0.05),而饲料系数显著低于对照组(P0.05);C/N16和C/N20处理组中团头鲂肠道的蛋白酶活性和淀粉酶活性显著高于对照组(P0.05);而各实验组中团头鲂肠道的脂肪酶活性没有显著性差异;C/N16和C/N20处理组中团头鲂肝脏超氧化物歧化酶、碱性磷酸酶和溶菌酶活性显著高于对照组(P0.05)。研究表明,生物絮团技术应用于团头鲂养殖适宜的C/N应不低于16,该条件下形成的生物絮团能有效提高团头鲂生长、消化酶和免疫相关酶活性。     

零换水条件下饲料蛋白水平对团头鲂幼鱼生长、消化酶活力和血清生化指标的影响

为探究零换水养殖水体中饲料蛋白水平对团头鲂(Megalobrama amblycephala)幼鱼生长、消化酶活力和血清生化指标的影响,实验设计投喂4个不同蛋白水平的生物絮团试验组(BF-20%、BF-25%、BF-30%、BF-35%)以及1个投喂35%蛋白的循环水对照组,每个处理组设置3重复,每个养殖桶放养初始体重为(6.06±0.01)g团头鲂幼鱼25尾,养殖周期为6周。结果表明:(1)形成的生物絮团可以有效地调节水质,降低水体中的氨氮、硝酸盐氮和亚硝酸盐氮水平;(2)BF-30%和BF-35%组和对照组的终末体重、增重率和饲料系数差异不显著(P0.05),但其显著高于BF-20%组(P0.05),各处理组间的存活率没有显著性差异(P0.05);(3)BF-25%组、BF-30%组和BF-35%组的肠道、肝脏超氧化物歧化酶和谷胱甘肽过氧化物酶活性显著高于对照组和BF-20%组(P0.05),而BF-20%组的肝脏丙二醛含量显著高于其他处理组(P0.05);BF-25%组、BF-30%组和BF-35%组的血清碱性磷酸酶和溶菌酶活性显著高于对照组和BF-20%组(P0.05),而其血清中总蛋白含量、白蛋白含量、谷丙转氨酶和谷草转氨酶活力差异不显著(P0.05)。研究表明,零换水条件下饲料中35%蛋白水平减少至25%并不影响团头鲂幼鱼的生长和非特异性免疫力。     

不同养殖模式对虎龙杂交斑生长、消化酶活性及非特异性免疫的影响

为研究不同养殖模式对虎龙杂交斑(E.fuscoguttatus♀×E.lanceolatus♂)生长、消化酶活性及非特异性免疫的影响,选取初始体质量为(143.22±12.48)g的健康虎龙杂交斑鱼种360尾,随机分为3个组:工厂化循环水养殖模式(A组)、池塘网箱养殖模式(B组)、流水养殖模式(C组),进行为期56 d的养殖试验。结果显示:B组的存活率、体质量增长率和特定生长率显著高于A组和C组(P0.05),生长激素(GH)、胰岛素样生长因子-Ⅰ(IGF-Ⅰ)水平也显著高于A组和C组(P0.05),其饲料系数(FCR)显著低于A组和C组(P0.05);B组肠道的淀粉酶和脂肪酶的活性显著高于A组和C组(P0.05),B组和C组的蛋白酶活性显著高于A组(P0.05);B组血清丙二醛含量显著高于A组和C组(P0.05),A组血清溶菌酶含量和超氧化岐化酶活性显著高于B组和C组(P0.05)。研究表明:池塘网箱养殖模式能更好地促进虎龙杂交斑的生长和消化,工厂化养殖模式则更有利于提高鱼的免疫力。     

不同蛋氨酸剂型对中华鳖生长性能和肝脏基因表达的影响

为了研究在低鱼粉含量饲料中分别添加不同蛋氨酸剂型对中华鳖生长性能、消化酶活性、肝脏相关基因表达的影响,以初始体质量为(3.48±0.03)g的中华鳖(Pelodiscus sinensis)稚鳖为试验对象,配制5组饲料,其中正对照组(T1)饲料蛋白的质量分数为46%,添加鱼粉的质量分数为46%,负对照组(T2)饲料蛋白质量分数为43%,添加鱼粉的质量分数为23%,在T2饲料的基础上分别添加0.1%DL-蛋氨酸(T3)、0.2%包膜蛋氨酸(T4)和0.125%羟基蛋氨酸钙(T5),开展持续8周的养殖试验。结果显示,T2组的特定生长率显著低于T1组(P0.05),摄食添加不同蛋氨酸剂型饲料的试验组,中华鳖的特定生长率均显著高于T2组,其中T3、T5组与T1组无显著差异(P0.05)。添加蛋氨酸可显著提高试验鳖的胃淀粉酶活性(P0.05),T3和T5组的胃蛋白酶活性以及T3组的脂肪酶活性相对T2组均显著提高(P0.05)。添加蛋氨酸显著提高了中华鳖肝脏GHR和IGF-I基因mRNA的表达量(P0.05),T3和T5组的IGF-ⅡmRNA相对表达量也显著高于T2组(P0.05)。T5组的HSP70 mRNA相对表达量显著高于T2和T4组(P0.05),T2组的HSP90 mRNA相对表达量显著低于除T4组以外的其它各组(P0.05)。结果表明,在低鱼粉含量的饲料中补充不同剂型蛋氨酸对中华鳖幼鳖的生长性能有不同程度的促进作用,其中补充0.1%的DL-蛋氨酸和0.125%的羟基蛋氨酸钙可达到T1组的养殖效果。     

乳化剂对异育银鲫消化酶及血清指标的影响

选取8000尾异育银鲫,随机分为2个处理(对照组和试验组),每处理2重复,每重复2000尾,分别饲喂含乳化剂0%和0.05%的基础日粮,试验期为62 d。试验结果表明,饲养31 d后,添加乳化剂使异育银鲫的肝胰脏胰蛋白酶的活性显著提高(P<0.01),肠道胰蛋白酶活性也表现出升高的趋势(P=0.08);血清中总蛋白、球蛋白、葡萄糖和甘油三酯的含量也显著提高(P<0.01)。饲养62 d后,添加乳化剂使异育银鲫肠道淀粉酶活性显著高于对照组(P<0.05);血清中葡萄糖和甘油三酯的含量显著高于对照组(P<0.01)。试验结果表明,饲料中添加乳化剂能够提高异育银鲫的肠道和肝胰脏消化酶的活性,促进脂肪代谢和蛋白质沉积。     

不同养殖模式对尼罗罗非鱼生长、免疫性能及水质的影响

为探讨循环水养殖模式(RAS组)和池塘养殖模式(池塘组)对尼罗罗非鱼(Oreochromis niloticus)生长、免疫性能及水质影响的变化规律,开展了为期8周的养殖试验。结果表明:(1)RAS组中鱼体增重率(WGR)[(597.36±169.79)%]显著高于池塘组[(470.98±142.99)%](P0.05),饲料转化率(FCR)[(1.17±0.02)%]却显著低于池塘养殖系统[(1.47±0.03)%](P0.05)。特定生长率(SGR)和肥满度(CF)也高于池塘养殖系统,但差异不显著(P0.05);(2)在第4周和第8周分别测定了鱼体胃和肠中消化酶活性,第4周时,RAS组中肠蛋白酶活性值[(1 387.56±278.43)μg/(g·min)]显著高于池塘组[(1 129.99±382.67)μg/(g·min)](P0.05);第8周时,肠脂肪酶活性[(18.11±4.28)μg/(g·min)]显著高于池塘组[(12.89±8.00)μg/(g·min)](P0.05);(3)在第4周和第8周分别测定了鱼体肝胰脏、头肾和血清中碱性磷酸酶(ALP)、溶菌酶(LSZ)和总超氧化物歧化酶(T-SOD)活性,整体表现为RAS组免疫性能低于池塘组;(4)每周测定水体氨态氮和亚硝态氮含量显示,RAS组氨态氮维持在0.0067~0.0212 mg/L,亚硝态氮含量为0.0027~0.0087 mg/L,均低于池塘组。     

Effect of substitution of dietary fish meal by soybean meal on different sizes of gibel carp (Carassius auratus gibelio): digestive enzyme gene expressions and activities,and intestinal and hepatic histology

A 7‐week growth trial was conducted to evaluate the effects of dietary soybean meal (SBM) on digestive enzyme activity of intestinal mucosa, mRNA levels of digestive enzymes in hepatopancreas, and the mid‐intestinal and hepatopancreas histology of gibel carp CAS III (Carassius auratus gibelio). Four different growth phases of gibel carp (initial body weight: fry, 0.8 g; juvenile, 5.0 g; 1‐year‐old, 62.7 g; and broodstock, 135.6 g) were tested. Seven isonitrogenous and iso‐energetic diets were formulated to contain different SBM replacement levels (0%, 20%, 40%, 60%, 80% and 100% of dietary fish meal protein), and another diet (SBMAA) contained all SBM protein and supplied crystalline amino acids. The results showed that the activities of mid‐intestine trypsin, α‐amylase and gamma glutamyl transpeptidase reduced with increased dietary SBM, while the chymotrypsin activity increased first and then decreased. The ultrastructures of intestinal epithelial cells and hepatopancreas cells in fry and broodstock fish were distinctly affected by 200 g kg^‐1 dietary SBM. Supplementation of dietary amino acid to the highest replacement groups was not sufficient to improve digestive and absorptive capacities and growth performance. Gibel carp may be adapted to dietary SBM through increase in gene expression of hepatopancreas digestive enzymes and has potential to utilize proceeded SBM as feedstuffs.     

Distribution of the introduced cyprinid herpesvirus 3 in a wild population of common carp, Cyprinus carpio L.

Cyprinid herpesvirus 3 (CyHV-3), which causes a lethal disease in common carp, Cyprinus carpio L., and koi, C. carpio koi , first occurred in Lake Biwa, Japan in 2004. To elucidate distribution of CyHV-3 in a wild common carp population, we conducted a PCR survey of CyHV-3 among such fish in Lake Biwa in 2006. Only 6% (1/18) of the common carp smaller than 300 mm were positive with PCR, whereas 31% (18/58) of fish larger than 300 mm were positive. To evaluate their past exposure to CyHV-3 infection based on the presence of antibodies, we also measured the levels of serum anti-CyHV-3 antibodies in the carp, using an enzyme-linked immunosorbent assay. None (0/26) of the fish smaller than 300 mm was positive for the antibodies, whereas 54% (33/61) of fish larger than 300 mm were positive. Of the antibody-positive individuals, 44% (14/32) were also positive by PCR strongly suggesting that wild common carp that survived infection become CyHV-3 carriers. Five individuals were positive by PCR but negative for antibodies indicating that their infection with CyHV-3 had occurred recently. These results suggest that transmission of CyHV-3 from carriers to naïve common carp is still occurring in Lake Biwa.     

鲤疱疹病毒Ⅱ型的理化及生物学特性和超微形态发生

为了查明鲤疱疹病毒Ⅱ型(Cyprinid herpesvirus 2,Cy HV-2)的理化与生物学特性以及病毒在细胞内的超微形态发生过程,利用新建立的对Cy HV-2敏感的异育银鲫脑组织细胞系(Gi CB),对Cy HV-2的理化及生物学特性进行了详细研究,比较了不同来源鱼类细胞系对Cy HV-2感染的敏感性,并对体外培养细胞中Cy HV-2病毒粒子及其超微形态发生过程进行了电镜观察。结果显示,Cy HV-2对热、酸、碱、有机溶剂和冻融敏感;常用鱼类细胞系EPC、RTG-2、Koi-Fin、CIK、CCK、PF-Fin对Cy HV-2的感染不敏感,特异性巢式PCR检测盲传至第7代Cy HV-2细胞培养物,结果均为阴性;Cy HV-2在Gi CB细胞中的增殖动态研究结果表明:病毒感染细胞经过12 h的隐晦期,24 h开始进入对数生长期,96 h病毒滴度达到最高值(107.52±0.26 TCID50/m L),然后进入平台期;透射电子显微镜观察结果显示,Cy HV-2感染细胞可分为吸附与侵入、复制与装配、成熟与释放3个主要过程,病毒进入对数生长期后,被感染细胞内可见形态典型的疱疹病毒颗粒。     

Effect of different water biofloc contents on the growth and immune response of gibel carp cultured in zero water exchange and no feed addition system

This study evaluated the growth and immune response of gibel carp (Carassius auratus gibelio) cultured under no feed addition biofloc technology (BFT) system at different total suspended solid (TSS) concentrations (10, 300, 600, 800 and 1,000 mg/L for group BF0‐NF, BF300‐NF, BF600‐NF, BF800‐NF and BF1000‐NF) for 30 days. The results demonstrated that bioflocs contained rich nutrients, and gibel carp eaten bioflocs showed higher weight gain, specific growth and survival. Digestive enzyme activities such as pepsin and amylase increased significantly in BF300/600/800/1000‐NF groups than those in BF0‐NF group. Antioxidant response including superoxide dismutase and total antioxidant capacity in serum and skin mucus was also enhanced significantly (p < .05). In addition, six immune‐related genes were examined by RT‐qPCR. Compared with BF0‐NF group, expression levels of immune genes intelectin (ITLN), dual specificity phosphatase 1 (DUSP 1), keratin 8 (KRT 8), myeloid‐specific‐peroxidase (MPO), c‐type lysozyme (c‐lys) and interleukin‐11 (IL‐11) were up‐regulated by 78.1‐, 23.9‐, 13.8‐, 138.8‐, 401.8‐ and 91.1‐fold, respectively. The highest expression values were observed at TSS of 600–800 mg/L. This study suggested that bioflocs can be uptaken by gibel carp as a food source, and have a potential to be used as a supplemental food for aquaculture.     

Microsatellite marker isolation and cultured strain identification in <Emphasis Type="Italic">Carassius auratus gibelio</Emphasis>

Microsatellites have become the preferred molecular markers for strain selection and genetic breeding in fish. In this study a total of 105 microsatellites were isolated and identified in gibel carp (Carassius auratus gibelio) by microsatellite sequence searches in GenBank and other databases and by screening and sequencing of positive clones from the genomic library enriched for AG and GATA repeats. Moreover, nineteen microsatellites were randomly selected to design locus-specific primer pairs, and these were successfully used to identify and discriminate different cultured strains of gibel carp including strains A, D, L, and F. Three different types of microsatellite pattern were distinguished by the number and length of fragments amplified from the 19 primer pairs, and some microsatellite primer pairs were found to produce different microsatellite patterns among strains and strain-specific fragments. In addition, some duplicated alleles were also detected in two microsatellite patterns. Therefore, the current study provides direct molecular markers to discriminate among different cultured strains for selective breeding and aquaculture practice of gibel carp.     

Production performance of largemouth bass Micropterus salmoides and water quality variation in monoculture,polyculture and integrated culture

A 74‐day experiment was conducted to evaluate the production performance and water quality variation in three types of farming system for largemouth bass Micropterus salmoides. The tested aquaculture models included monoculture of largemouth bass (MC), polyculture of largemouth bass, gibel carp Carassius auratus gibelio and silver carp Hypophthalmichthys molitrix (PC), and integrated culture of largemouth bass, gibel carp, silver carp and freshwater pearl mussel Hyriopsis cumingii (IC). The ratio of largemouth bass, gibel carp and silver carp was 30:2:1 in the PC model, and the ratio of largemouth bass, gibel carp, silver carp and mussel was 30:2:1:5 in the IC model. The largemouth bass were fed with formulated feed twice daily. No significant differences were found in weight gain and yield of largemouth bass, total fish yield, nitrogen (N) and phosphorus (P) utilization efficiencies, N and P wastes, pH, nitrite, nitrate, reactive phosphate, total nitrogen, total phosphorus, total organic carbon, chemical oxygen demand, 5‐day biochemical oxygen demand, chlorophyll a, primary productivity among the MC, PC and IC models. The ammonia was lower, while the dissolved oxygen was higher in the PC tanks than in the MC tanks. These results suggest that the environment situation was better in the PC tanks relative to that in the MC tanks. The present study reveals that the PC model should be a way to optimize the aquaculture model for commercial largemouth bass farming.     

鲤疱疹病毒Ⅱ型ORF66基因DNA疫苗载体的构建及其免疫效果

鲤疱疹病毒Ⅱ型(CyHV-2)能够引起鲫大量死亡,严重威胁我国水产养殖业的健康发展,目前,针对CyHV-2尚无有效的商业化疫苗或治疗措施。为构建CyHV-2DNA疫苗,本研究将其衣壳蛋白ORF66编码基因克隆至pVAX1真核表达载体上,构建重组质粒pVAX-ORF66。此外,在BL21(DE3)pLysS中对ORF66蛋白进行了原核表达,表达蛋白经纯化后免疫新西兰白兔制备了ORF66特异性抗体。酶联免疫吸附(ELISA)检测显示,制备抗体效价达1∶20000以上。将pVAX-ORF66质粒转染金鱼脑细胞系(GFB),利用制备的ORF66抗体进行间接免疫荧光(IFA)检测,结果显示,ORF66蛋白可以在细胞中大量表达,且主要定位于细胞质中。将pVAX-ORF66质粒肌肉注射鲫后进行CyHV-2免疫保护实验,结果表明,其相对免疫保护率达55.6%。本研究针对CyHV-2构建了一种制备简单、成本低廉的DNA疫苗,为鲫造血器官坏死病的免疫预防及感染分子机制研究奠定了前期实验基础。     

Optimization of fish to mussel stocking ratio: Development of a state-of-art pearl production mode through fish–mussel integration

Integrated aquaculture has been widely used for pearl production in the freshwater pearl mussel Hyriopsis cumingii farming in China, but the production technology has not reached the state of the art. This study explored the optimal stocking ratio of fish to mussel (fish–mussel) through a 90-day experiment conducted in land-based enclosures. The integrated system included pearl mussel, grass carp, gibel carp, silver carp and bighead carp, with four fish–mussel stocking ratios by number: 1:1 (R1), 2:1 (R2), 3:1 (R3) and 4:1 (R4). The pearl yield was higher in the R2 enclosures than in the R1 and R4 enclosures, whereas the fish yield was higher in the R3 and R4 enclosures than in the R1 and R2 enclosures. The phosphorus (P) utilization efficiency was higher in the R2, R3 and R4 enclosures than in the R1 enclosures. The wastes of nitrogen (N) and P enhanced with the increase of fish–mussel ratio. Regression analyses indicated that the fish–mussel ratio was 2.3:1 for the maximal pearl yield, and 3.6:1 for the maximal fish yield, and 1.6–2.3:1 for the minimal N waste, and 1.9–2.9:1 for the minimal P waste. This study indicated that the suitable fish–mussel stocking ratio was 2:1 in the integrated culture of H. cumingii, grass carp, gibel carp, silver carp and bighead carp.