Formulated diet alters gut microbiota compositions in marine fish Nibea coibor and Nibea diacanthus

Nibea, an economical marine fish, is generally fed on trash fish (the low‐value fish), which can cause high feed costs and waste pollutions in high‐density aquaculture. To assess the effect of formulated diet on the gut microbiota in Nibea coibor and Nibea diacanthus, denaturing gradient gel electrophoresis, clone libraries analysis and Illumina sequencing of the 16S ribosomal RNA gene were used in this study. Two Nibea fishes were both dominated by Gammaproteobacteria (especially Photobacterium) and shared a set of gut microbiota, including Bacilli, Mollicutes, Alphaproteobacteria and Fusobacteriia. Statistical analyses revealed that formulated diet led to lower feed conversion ratio (p < 0.001), lower abundance of Vibrio (p = 0.040) and infectious diseases pathways (p = 0.001), higher abundance of polysaccharide‐degrading bacterium Cellvibrio (p = 0.006) in two Nibea species, with higher weight gain rate (p = 0.023) and microbial diversity (Shannon, p = 0.049 and Simpson, p = 0.044) and more carbohydrate metabolism (p = 0.020) observed in N. coibor. The distribution and correlation network of 17 potential short‐chain fatty acid producing bacteria were obtained and visualized in all treatment groups. The results reveal that formulated diet has beneficial effects on the gut microbial ecology in two Nibea fishes, which suggests the possibility of replacing trash fish diet with formulated diet in Nibea aquaculture.     

Effect of hydroxyproline supplementation on growth performance,body composition,amino acid profiles,blood‐biochemistry and collagen synthesis of juvenile chu's croaker (Nibea coibor)

Hydroxyproline (Hyp) may play an important role in collagen metabolism, as it is almost exclusively found in collagen. To examine this possibility, a 56‐day feeding trial was conducted to evaluate the effect of graded supplementaries of Hyp (0, 2.5, 5, 7.5, 10 and 12.5 g/kg dry diet weight) on growth performance, feed utilization, biochemical composition and collagen synthesis of chu's croaker, Nibea coibor (13.6 ± 0.28 g). The growth performance and feed utilization were improved with increased levels of dietary Hyp, reaching a maximum at 5 g/kg (diet H2) followed by a slight decrease. Polynomial regression analysis showed that the maximum specific growth rate (SGR) was attained at 7.177 g/kg dietary Hyp. Increasing levels of Hyp did not affect body composition of juvenile Nibea coibor. However, dietary Hyp supplementation significantly affected the levels of Leu, Ile, Met, Lys, Ser, Glu and Pro in muscle and the levels of Met, Lys, Glu, Pro and Gly in swim bladder. The serum calcium (SC) level decreased significantly with increasing levels of dietary Hyp. However, the triglycerides (TG) and total cholesterol (T‐CH) levels in serum and liver were relatively stable. Importantly, the acid‐soluble collagen (ASC) and total collagen (TC) levels in swim bladder increased significantly initially and then decreased after reaching a maximum. Polynomial regression analysis revealed that the maximum TC level in swim bladder was attained at 9.736 g/kg of dietary Hyp. Collectively, these results suggest that the appropriate provision of hydroxyproline is essential for maximal collagen synthesis and growth.     

日本黄姑鱼血细胞发生的观察

用Wright’sGiemsa混合染料对日本黄姑鱼外周血涂片以及头肾、肾脏、肝脏和脾脏组织印片染色,观察其中各种血细胞的大小、形态特征,研究了日本黄姑鱼血细胞的发生发育模式。观察发现,在外周血涂片中,除红细胞外,还观察到多种白细胞,主要有单核细胞、嗜中性细胞、嗜酸性细胞和淋巴细胞,而嗜碱性细胞没有被发现。红细胞、淋巴细胞和单核细胞的发生主要在肾脏和头肾,其次是脾脏;粒细胞的发生主要在头肾和脾脏。红细胞的发育经历3个阶段:原红细胞、幼红细胞和成熟红细胞阶段。红细胞在成熟过程中,细胞核的体积逐渐变小,细胞与细胞核的体积比由大变小然后再变大。成熟红细胞除了由幼稚细胞发育而来以外,还可以进行一分为二的方式进行分裂;粒细胞的发育经历5个阶段:原粒细胞、早幼粒细胞、中幼粒细胞、晚幼粒细胞和成熟粒细胞;淋巴细胞经历了3个阶段:原淋巴细胞、幼淋巴细胞和成熟淋巴细胞;单核细胞的发育和淋巴细胞相似,也经历了3个阶段:原单核细胞、幼单核细胞和成熟单核细胞。     

浅色黄姑鱼鳃结构及其呼吸面积的研究

应用扫描电镜技术对体重范围在4.75～35.51g的浅色黄姑鱼（Nibea coibor）鳃结构及鳃小片进行了观察与呼吸面积计算,研究体重、体长与呼吸面积之间的相关关系。结果表明,浅色黄姑鱼的平均相对呼吸面积为4.01±1.62cm^2·g^-1。体重与一侧鳃丝总数的相关性N=28.81＋158.45lnW（R^2=0.9670,P〈0.01）较体长与一侧鳃丝总数的相关性N=17.31L^1.4542（R^2=0.8241,P〈0.01）更为显著。单位mm鳃小片的数目随着体重的不断增加显著减少,N=49.971W^-0.3909（P〈0.01）。单个鳃小片的面积随体重的增大而显著增加,a=0.3199W^0.2528（P〈0.01）。总呼吸面积随体重的增大而显著增加,A=11.559W^0.7109（P〈0.01）。相对呼吸面积随体重的增大而显著减小,RA=11.561W^-0.2904（P〈0.01）。     

日本黄姑鱼养殖试验初报

从国外首次引入日本黄姑鱼受精卵，育成苗种后进行海水池塘及网箱的养殖试验，并进行越冬试验及亲鱼的继续培育。结果表明：（1）经5.5 个月的池塘养殖，个体平均体长达31.1cm、体重515.0g，养殖成活率为82.85%。（2）经5.5个月的网箱养殖，个体平均体长达28.1cm、体重438.8g，养殖成活率为81.66%。（3）日本黄姑鱼养殖水温下限为5.5-6.0℃。（4）经25个月的池塘养殖、亲鱼平均体长达54.0cm、体重2700.0g。     

饥饿和再投喂对日本黄姑鱼代谢率和消化器官组织学的影响

采用封闭式呼吸仪测定日本黄姑鱼(8.14±1.07 g)在饥饿和恢复投喂过程中的耗氧率;用石蜡切片法观察和分析饥饿和再投喂对日本黄姑鱼消化器官形态结构和组织学的影响。结果表明:随着饥饿时间的延长,其耗氧率逐渐降低;恢复投喂之后,其耗氧率开始逐步上升。从组织结构看,食道无明显变化,而胃、肠和肝胰脏则随着饥饿时间的延长出现不同程度的损伤;饥饿5 d组无明显变化,而饥饿15 d组变化明显,表现为胃腺细胞收缩,皱壁和上皮细胞高度减少,分泌颗粒减少,肠胃绒毛退化,肝组织致密,肝细胞内脂滴减少,胰腺泡缩小,排列不规则;饥饿10 d组介于饥饿5 d组和饥饿15 d组之间,再投喂各主要结构均有恢复。     

舟山近海黄姑鱼胚胎发育及前期仔鱼形态的观察

通过对黄姑鱼胚胎发育全过程的连续观察,描述了黄姑鱼胚胎发育的特征、发育速度及前期仔鱼的形态特征等。观察结果表明,黄姑鱼的胚胎发育与其它硬骨鱼浮性卵鱼类基本相似,属端黄卵,盘状分裂。在水温22.0~22.5℃、盐度26.5的条件下,黄姑鱼受精卵经24h15m in孵出仔鱼。初孵仔鱼全长1.75±0.09mm,孵出后约50h,卵黄囊完全被吸收。     

浅色黄姑鱼线粒体16S rRNA基因片段序列特征分析

PCR扩增100个浅色黄姑鱼个体的线粒体16SrRNA基因片段序列,得到大约620bp的扩增产物。将其中5个个体的扩增产物进行测序和同源比对,得到468bp可供比对分析的片段。比对结果表明,5条序列包括两个单倍型,两个单倍型之间有1个碱基突变。PCR-RFLP分析结果显示,两个种群的100个样品中98%的个体为其中一种单倍型,只有2%的个体呈另一种单倍型,表明这两个种群的遗传多样性较低。两个单倍型平均碱基组成为:T22.0%,C26.3%,A29.8%,G21.9%,GC含量平均为48.2%。与GenBank中石首鱼科7属9种的11条同源序列比对,得到429个比对位点,其中包括69个简约信息位点、55个单突变子和16个插入/缺失位点。聚类分析显示,浅色黄姑鱼与黄姑鱼亲缘关系较近,与形态分类相符。     

Hg↑（2+）、Zn↑（2+）、Cr↑（6+）对黄姑鱼幼鱼的急性致毒效应

2004年8月，在浙江省舟山市定海区小沙镇浙江华兴海水种苗有限公司以日龄为56d的黄姑鱼（Nibeaalbiflora）幼鱼作为实验动物，开展Hg^2＋、Zn^2＋、Cr^6＋对黄姑鱼幼鱼的急性毒性和加和等毒性强度联合毒性试验。结果表明，Hg^2＋对黄姑鱼幼鱼24h、48h、72h、96h的半致死质量浓度分别为0．7523mg／L、0．6616mg／L、0．5618mg／L、0．4959mg／L；Zn^2＋对黄姑鱼幼鱼24h、48h、72h、96h的半致死质量浓度分别为22．9908mg／L、18．4576mg／L、14．5306mg／L、11．5833mg／L；Cr^6＋对黄姑鱼幼鱼24h、48h、72h、96h的半致死质量浓度分别为18．7654mg／L、16．7415mg／L、15．4521mg／L、13．1276mg／L。黄姑鱼幼鱼对Cr6’、Zn^2＋、Hg^2＋96h安全质量浓度分别为1．31mg／L、1．16mg／L和0．050mg／L。各重金属离子毒性由大到小依次为Hg^2＋、Zn^2＋、Cr^6＋,Hg^2＋-Zn^2＋、Hg^2＋,Cr^6＋对黄姑鱼幼鱼的96h联合急性毒性表现为协同作用。Zn^2＋-Cr^6＋对黄姑鱼幼鱼的96h联合急性毒性表现为，低毒性强度的Zn^2＋对Cr^6＋具拮抗作用Cr^6＋毒性强度略高于Zn^2＋时表现为相互独立作用，当Zn^2＋毒性强度高于Cr^6＋时则表现为协同作用。并就Hg^2＋、Zn^2＋、Cr^6＋对黄姑鱼幼鱼的急性致毒效应特征，黄姑鱼幼鱼对Hg^2＋、Zn^2＋、Cr^6＋的安全浓度以及Cr^6＋-Zn^2＋、Cr^6＋-Hg^2＋与Zn^2＋-Hg^2＋对黄姑鱼幼鱼的联合毒性效应机理进行了探讨。     

海洋暖化和酸化对黄姑鱼早期生长发育的影响

以黄姑鱼(Nibea albiflora)为研究对象,根据IPCC 2013典型浓度排放路径(RCP)对2100年海洋温度和p H的预测值,分别选择减缓温室气体排放情景(RCP 2.6)和高温室气体排放情景(RCP8.5),设置温度和p H两个因素,在实验室条件下模拟气候变化,探究海洋暖化和酸化对黄姑鱼早期生长、发育的影响。结果显示,在基础温度24.0℃条件下,黄姑鱼进入稚鱼期的时间为25 d,高温处理(26.0℃~28.0℃)生长加快,仅为22 d,同时,随着温度升高(26.0℃、26.6℃和28.0℃),生长率逐渐增加(0.73、0.74和0.78 mm/d),并且各处理组之间生长率存在显著差异(P0.05),但死亡率在26.6℃最高,26.0℃最低。黄姑鱼死亡率在p H为7.80、7.73和7.49时分别为3.9%、19.4%和21.7%,各处理组死亡率差异显著(P0.05),但p H对黄姑鱼早期生长率影响不显著(P0.05)。通过逻辑斯蒂生长方程拟合,结合IPCC对2100年温度和p H的预测,到2100年,在RCP 2.6情景下,黄姑鱼早期生长率为0.71 mm/d,死亡率为31.1%;在RCP 8.5情景下,黄姑鱼生长率最高达到0.76 mm/d,死亡率为23.9%。在海洋酸化和暖化的共同作用下,黄姑鱼的早期补充将会受到影响。