草鱼胰岛素样生长因子1受体基因cDNA全序列的克隆及功能

本研究采用cDNA末端快速扩增法(RACE)克隆了草鱼(Ctenopharyngodon idellus)IGF-IR基因全长cDNA序列, 并对该基因在草鱼不同时期胚胎和成鱼不同组织中的表达进行了分析。序列分析表明, 草鱼IGF-IR基因cDNA序列全长5 741 bp, 包括5′端非翻译区822 bp, 3′端非翻译区581 bp, 开放阅读框4 338 bp, 共编码1 445个氨基酸。序列比对结果显示, 草鱼IGF-IR可能属于a型, 该基因编码的氨基酸序列与鲤(Cyprinus carpio)IGF-IRa、斑马鱼(Danio rerio)IGF-IRa和人类(Homo sapiens)IGF-IR的相似性分别为95%、93%和66%, 具有较高的同源性, 表明该基因在长期进化中具有较高的保守性。RT-PCR结果表明, 该基因从16 hpf(hours post fertilization)胚胎期到出苗期都有表达, 在成鱼大部分组织中均有表达。原位杂交结果显示, 草鱼IGF-IR mRNA在不同时期胚胎组织中广泛存在, 其中在脑部、脊索和尾部等生长旺盛组织的细胞中表达量较高。本研究为进一步探索草鱼IGF-IR基因在生长发育信号通路中的作用和育种提供了基础资料。

     

草鱼不同混养模式下围隔底泥反硝化、硝化和氨化速率

应用乙炔抑制法测定了草鱼(Ctenopharyngodon idellus)不同混养模式围隔底泥的硝化、反硝化和氨化速率,以探究草鱼不同混养模式对池塘底泥－水界面N元素动态变化的影响,并为草鱼养殖模式的优化提供必要的参考依据.混养组合分别为一元组（草鱼）、二元组（草鱼+鲢、草鱼+凡纳滨对虾）、三元组（草鱼+鲢+凡纳滨对虾,设2种放养比例）.结果显示:1）草鱼不同混养模式中底泥的反硝化速率范围为0～734.15 μmol/(m2·d),硝化速范围为0～1 209.20 mmol/(m2·d),化速率范围为0～41.25 mmol/(m2·d).2)草鱼不同混养模式底泥的反硝化速率较高,与很多河口和湖泊数值接近;在养殖中期,底泥的硝化速率很小甚至检测不到;底泥的氨化速率呈逐月递增趋势,以三元混养组最高.3）混养组中凡纳滨对虾(Litopenaeus vannamei)的放养密度越大,草鱼的放养密度越小,反硝化作用出现得越早,但反硝化速率很难保持开始的水平;反之,反硝化作用出现得较晚,并会随着养殖周期的推移迅速升高;放养种类越单一,底泥的硝化速率越低,且鲢(Hypophthalmichthys molitrix)放养密度越高,硝化速率越高.从养殖模式优化的角度来看,草鱼三元混养组要优于单养和二元混养组,但三元混养组中两种放养密度各有优劣,有待进一步优化以确定最佳放养比例.     

一个与大鲵蛙病毒 (ADRV) 感染相关基因 —6R的克隆、原核表达及其产物分离

在新近报道大鲵蛙病毒(Andriasda vidianus ranaviurs,ADRV)基因组的基础上,我们对ADRV 6R进行了克隆、原核表达及其产物的分离,这是一个经生物信息分析表明与病毒感染相关的功能基因。先经PCR扩增长到711 bp的ADRV6R,构建含扩增片段的克隆质粒p MD18-T-6R。再构建原核表达质粒p ET-32a-6R,进行测序,并与水生动物蛙病毒相关基因同源序列进行比对,该扩增片段与预期一致,其编码分子量约为27 k D、含236个氨基酸的蛋白,与水产动物病毒US22蛋白家族(US22 family protein)成员有很高的同源性。然后对p ET-32a-6R进行转化、诱导表达,并利用Ni-NTA His-Bind亲和层析及不同浓度咪唑洗脱液对表达产物进行分离、电泳分析,结果显示,获得与预测分子量相符的45 k D融合蛋白(27 k D目的蛋白与18 k D His标签)。这为后续制备抗体、并开展大鲵蛙病毒与宿主的相互作用研究提供了有用的实验材料。     

MEY = MSY

It has been generally accepted for more than half a century that the fishing sector stood to gain from managing fisheries at the effort level producing maximum economic yield (MEY) rather than at the higher effort level producing maximum sustainable yield (MSY). However, the acceptance is built on evaluating only the revenue and cost structure for the fishing fleet, not for the overall fishing sector including processing, distribution and marketing of fish products. Considering these links of the fish value chain moves the MEY-level closer to, but slightly below the MSY-level. For society as a whole, this means that MSY is the more appropriate target reference level.     

Dietary manganese requirement of juvenile hybrid grouper,Epinephelus lanceolatus × E. fuscoguttatus

An 8‐week feeding trial was conducted to estimate the optimum dietary manganese (Mn) requirement for juvenile hybrid grouper, Epinephelus lanceolatus × E. fuscoguttatus. The basal diet was formulated to contain 520 g/kg crude protein from casein and fishmeal. Manganese methionine was added to the basal diet at 0 (control group), 2.5, 5, 10, 20 and 40 mg Mn/kg diet providing 7.48, 10.34, 13.76, 19.72, 31.00 and 53.91 mg Mn/kg diet, respectively. Each diet was randomly fed to triplicate groups of juveniles, and each tank was stocked with 20 fish (initial weight, 60.06 ± 0.68 g). The manganese content in rearing water was monitored and kept below 0.01 mg/L. Results showed that the weight gain ratio (WGR), protein efficiency ratio (PER), specific growth rate (SGR), Mn contents in whole body, liver and vertebra, and activities of hepatic Mn superoxide dismutase (Mn‐SOD), total SOD (T‐SOD) and glutathione peroxidase (GSH‐PX) were significantly improved by dietary Mn supplementation (p < .05). However, dietary Mn did not affect arginase (DArg) activity. The highest feed conversion ratio (FCR) was observed in fish fed the basal diet (p < .05). No significant differences were found on the Cu and Zn contents in whole body by supplementing dietary Mn. Supplemented Mn in diets had significantly effect on liver and vertebral trace element deposition (p < .05). Fish fed the basal diet had the highest Fe and Zn contents in vertebra (p < .05). There were no significant differences on hepatic pyruvate decarboxylase (PDC) activity with supplemented Mn levels below 13.76 mg/kg. As biomarker of oxidative stress, malondialdehyde (MDA) content in liver was significantly higher in fish fed the basal diet (p < .05). Using the broken‐line models based on SGR, dietary Mn requirement of the juvenile hybrid grouper was estimated to be 12.70 mg/kg diet.     

秦皇岛海域野生牙鲆群体遗传多样性分析

用18对多态性微卫星标记对采自秦皇岛海域的90尾野生牙鲆(Paralichthys olivaceus)进行遗传分析。18个微卫星位点共检测出161个等位基因;各位点等位基因数为7~11个,平均为8.9;有效等位基因为3.7~8.0,平均有效等位基因为5.9;各个位点多态信息含量为0.69~0.86,平均值为0.80;Shannon多样性指数平均值为1.9;观测杂合度(Ho)值为0.33~0.87,平均值为0.64;期望杂合度为0.74~0.89,平均期望杂合度(He)为0.84。χ2检验表明,18个位点中有9个的等位基因分布偏离了哈迪温伯格平衡(P<0.05)。研究结果表明,秦皇岛海域野生牙鲆群体遗传信息含量丰富、等位基因分布均匀,遗传多样性较高,但是,存在Hardy-Weinberg不平衡现象。     

Apparent digestibility of Asian carp‐ and common carp‐derived fish meals in feeds for hybrid striped bass Morone saxatilis ♀ × M. chrysops ♂ and rainbow trout Oncorhynchus mykiss

Apparent digestibility coefficients (ADCs) of nutrients (crude protein, amino acids, crude lipid, fatty acids, and minerals) were determined for fish meals derived from menhaden, Asian carp (combination of silver and bighead carps), and common carp in feeds for hybrid striped bass and rainbow trout. Extruded test diets were formulated to contain a 70 : 30 mixture of reference diet and test ingredient with yttrium oxide (1 g kg^?1) serving as the inert marker. Diets were randomly assigned to triplicate tanks and fish were fed once per day at 2% body weight. Fecal samples were collected by manual stripping. The ADCs were calculated according to standard procedures. The composition and digestibility of Asian carp and common carp meals was broadly similar to menhaden meal. Protein digestibility ranged from 86.5% (Asian carp meal) to 93.1% (common carp meal). Lipid was highly digestible with ADCs >100% for all ingredients. Although the Asian carp meal was less digestible than the other two fish meals, it was still a highly digestible ingredient. Our data suggest that fish meals derived from Asian or common carp would be valuable feedstuffs in diets for hybrid striped bass, rainbow trout, and possibly other cultured fishes.     

Molecular cloning,characterization, tissue expression and nutritional regulation of O‐GlcNAc transferase gene in hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂)

O‐GlcNAc transferase gene (OGT) was considered as the sole rate‐limiting enzyme in the O‐GlcNAc modification. In the present study, the OGT gene of hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) was cloned and characterized, and its expression in response to dietary carbohydrate level and acute glucose treatment was investigated. The full‐length of OGT (GenBank accession no. KY656469 ) was 4,063 bp, including a 302 bp 5′untranslated terminal region (UTR), a 3,165 bp coding region that encoded 1,054 amino acids residues and a 596 bp 3′ UTR. The highly conservation of OGT gene between fish and mammals was also observed through multiple sequences alignment and phylogenetic analysis. O‐GlcNAc transferase gene was ubiquitously expressed in all detected tissues with highest expressions in brain and liver, to a lesser degree, in eye, heart, kidney and intestine. The increasing dietary carbohydrate from 8.02% to 16.08% had no significant effect on the mRNA expression of OGT. However, the expression of OGT was slightly elevated at 6 hr post‐glucose injection, and the elevation became significant at 24 hr time‐point. These data may enhance our understanding on the nutritional regulation of OGT and O‐GlcNAc modification in fish species.     

Dietary arginine requirement of juvenile hybrid sturgeon (Acipenser schrenckii♀ × Acipenser baerii♂)

An 8‐week feeding trial was conducted to evaluate the dietary arginine requirement of juvenile hybrid sturgeon. Seven isonitrogenous and isolipidic diets were formulated to contain graded levels of dietary arginine ranging from 1.74% to 3.54% (dry weight). The results indicated that the fish fed with 1.76% arginine diet had lower specific growth rate (SGR) and feed efficiency ratio (FER) than the fish fed the 2.64% to 3.24% arginine diets (p < .05), and the fish fed the 2.64% arginine diet presented the highest SGR. The fish fed with 1.76% arginine diet had lower whole‐body crude protein content than the fish fed the 2.64% or 2.93% arginine diets (p < .05). Compared with the 1.76% arginine diet, 2.36% to 2.93% arginine diets significantly increased the total amino acid (TAA) concentration and total nonessential amino acid (TNEAA) of carcasses. The fish fed with 1.76% arginine diet had lower activity of nitric oxide synthase (T‐NOS) and content of nitric oxide (NO) than the fish fed the 2.05% to 2.93% arginine diets in the liver. The fish fed with 1.76%, 2.05% or 2.36% arginine diets had lower gene expression of growth hormone (GH) and insulin‐like growth factor I (IGF‐I) in the liver than the fish fed the 2.64% to 3.53% arginine diets in the liver (p < .05). A broken‐line analysis between SGR against the dietary arginine levels provided estimates for the optimal dietary arginine requirement of 2.47% corresponding to 6.18% of the dietary protein on a dry weight basis.     

充气培养对半叶马尾藻生长﹑营养盐吸收和生化组成的影响

将半叶马尾藻(Sargassum hemiphyllum)置于实验室球形培养瓶内培养15 d,设置5个充气速率(50 m L/min、100 m L/min、200 m L/min、400 m L/min和800 m L/min)为实验组,静水培养(0)为对照组。每隔5 d测定藻体的生长速率和营养盐吸收速率,培养实验结束时,测定藻体的色素含量、可溶性蛋白和可溶性糖含量。结果表明:充气速率和培养时间均对半叶马尾藻的生长和PO34--P的吸收具有显著影响,而对NO3--N来说,仅培养时间对其吸收影响显著。培养第1天时,充气可显著促进营养盐吸收,最大充气速率(800 m L/min)培养的半叶马尾藻对NO3--N和PO34--P的吸收速率分别比静水培养的马尾藻高133.77%和89.51%,生长速率也比静水培养的马尾藻高95.04%。随着培养时间的延长,充气速率对半叶马尾藻的生长和营养盐吸收的促进作用逐渐减弱。在培养第15天时,充气速率为800 m L/min条件下的半叶马尾藻对NO3--N的吸收速率比静水培养的马尾藻低24.87%,对PO34--P的吸收速率仅高出静水培养下的46.03%,而生长比静水培养的马尾藻高出75.16%。同时,除了可溶性蛋白含量,高充气速率可抑制叶绿素a(Chla)、类胡萝卜素(Car)和可溶性糖合成。因此,水体运动是通过对营养盐吸收及其他生理过程产生的作用间接影响半叶马尾藻的生长,而不是直接影响半叶马尾藻生长和产量的最重要因素。     

Hypoxia affects performance traits and body composition of juvenile hybrid striped bass (Morone chrysops × M. saxatilis)

Performance traits and body composition of juvenile hybrid striped bass (Morone chrysops × M. saxatilis) in response to hypoxia were evaluated in replicate tanks maintained at constant dissolved oxygen concentrations that averaged 23.0 ± 2.3%, 39.7 ± 3.0% and 105.5 ± 9.5% dissolved oxygen saturation. Fish were fed a commercially formulated feed daily to apparent satiation. Total feed intake and fish growth and yield increased linearly in response to increased dissolved oxygen concentration. Nutrient utilization was reduced significantly only at the greatest level of hypoxia. With the exception of whole body protein content, whole body compositional indices and nutrient retention efficiencies were linearly related to dissolved oxygen concentration. Results demonstrate that as hypoxia becomes more severe, juvenile hybrid striped bass feed intake is reduced, which affects growth and nutrient retention.     

Fishmeal replacement by soybean,rapeseed and cottonseed meals in hybrid sturgeon Acipenser baerii ♀ × Acipenser schrenckii ♂

A 12‐week feeding trial was conducted to evaluate the effects of replacing fishmeal (FM) with soybean meal (SBM), rapeseed meal (RM) and cottonseed meal (CSM) on growth, feed utilization and body composition of juvenile hybrid sturgeon Acipenser baerii ♀ × Acipenser schrenckii ♂ (initial body weight, 8.63 ± 0.24 g). Five isonitrogenous and isoenergetic diets were formulated as follows: a control diet (FM60) containing 600 g/kg FM and four other diets (FM45, FM30, FM15 and FM0 containing 450, 300, 150 and 0 g/kg FM, respectively) where protein from FM was substituted by a mixture of SBM, RM and CSM. Fish fed FM0 and FM15 had poorer growth performance, feed utilization, apparent digestibility coefficients of dry matter, protein, lipid and gross energy, and fed FM0 had poorer hepatosomatic index and survival compared with the fish fed FM60. The whole body lipid in fish fed FM0 was significantly higher than that in fish fed FM60 and FM15. This study indicates that 300 g/kg of FM can be replaced with a mixture of SBM, RM and CSM in the diet of juvenile hybrid sturgeon without compromising growth performance, feed utilization and body composition.     

Characterization of 38 kDa and 42 kDa chitinase isozymes from the liver of Japanese common squid Todarodes pacificus

ABSTRACT: Characterization was investigated on the 38 kDa and 42 kDa chitinase (EC3.2.1.14) isozymes from the liver of Japanese common squid Todarodes pacificus . Optimum pH toward colloidal chitin was observed at pH 3.0 for the 38 kDa chitinase, and pH 3.0 and 9.0 for the 42 kDa chitinase. K _m and k _cat of the 38 kDa and 42 kDa chitinases toward a longer substrate, glycol chitin, were 0.071 mg/mL and 1.22/s, and 0.074 mg/mL and 0.196/s, respectively. Alternatively, strong substrate inhibition of both chitinases were observed toward a short substrate, N -acetylchitopentaose (GlcNAc₅). Both chitinases decomposed not only chitin but also chitosan (D. A. 95%). The cleavage pattern and reaction rate were investigated using N -acetylchitooligosaccharides (GlcNAc_n, n  = 2–6). Both chitinases hydrolyzed GlcNAc_n ( n  = 4,5, and 6). The release of GlcNAc was not observed. The speed of the reaction was observed to be in the following order: GlcNAc₄ > GlcNAc₅ > GlcNAc₆ for the 38 kDa chitinase, and GlcNAc₆ > GlcNAc₅ > GlcNAc₄ for the 42 kDa chitinase. Both the chitinases released p -nitrophenol from p -nitrophenyl GlcNAc_n ( n  = 2, 3, and 4). N-terminal amino acid sequences of the 38 kDa and 42 kDa chitinases were YLLSXYFTNWSQYRPGAGKYFPQNI and EYRKVXYYTNWSQYREVPAKFFPEN, respectively.     

Ploidy polymorphism and morphological variation among reciprocal hybrids of Pseudosciaena crocea (♀) × Miichthys miiuy (♂)

Diploid and triploid hybrids of large yellow croaker (Pseudosciaena crocea, Pc) and Miiuy croaker (Miichthys miiuy, Mm) were produced by interspecific hybridization. The result showed that Pc (♀) and Mm (♂) can be successfully crossed and hybrid offspring of normal hatch rate were produced. In contrast, a lower fertilize and hatch rate were produced for hybrids of Pc (♂) and Mm (♀), and the offspring died day 19 after hatching. To confirm the ploidy of hybrid offspring, we examined the karyotypes of offspring. The result of DNA relative content investigation and genetic relationship analysis demonstrated that the major hybrid offspring were hybrid diploid. In addition, a small number of hybrid were triploid and gynogenesis diploid. Morphological analysis based on the measurable traits data indicated the success of the interspecific crosses. The natural gynogenetic diploid large yellow croaker (PP), diploid hybrids (PM) and triploid hybrids (PPM) were closer to maternal parent (Pc) than paternal parent (Mm). This study provided genetic evidences of the chromosome, DNA content and morphological levels to support the successful establishment of the polyploidy hybrids of Pc (♀) and Mm (♂).     

Responses of hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii ♂) to oral administration of phosphorus

To better understand the phosphorus (P) utilization in hybrid sturgeon (Huso dauricus ♀ × Acipenser schrenckii ♂), a combined technique with oesophageal intubation, dorsal aorta cannulation and urinary catheterization was used in the present study. Groups of five hybrid sturgeon were orally administrated with graded doses of P (0, 50, 100 and 200 mg P/kg BW) in the form of monocalcium phosphate. The high dose of P (200 mg P/kg BW) caused a significant increase in plasma P concentration. Plasma calcium (Ca) concentrations were significantly increased in the fish treated with 0 and 100 mg P/kg BW. Both P and urea excretion showed increased changing pattern with increasing P intubation level and exhibited significant higher value in 200 mg P/kg BW group. A significant increase in Ca excretion was found in the group without P supplementation (0 mg P/kg BW). ALP and SOD activity in plasma was inhibited in fish at P‐deficient groups (0 and 50 mg P/kg BW). The data in the current study showed that hybrid sturgeon at 100 mg P/kg BW treatment exhibited better P and Ca utilizations with lower waste excretion, as well as favourable status of anti‐oxidation.     

Vaccination strategies with oral booster for surubim hybrid (Pseudoplatystoma corruscans × P. reticulatum) against haemorrhagic septicaemia

This study evaluated the efficiency of differently prepared vaccines against Aeromonas hydrophila in the hybrid surubim (Pseudoplatystoma corruscans × P. reticulatum). Survival and haemato‐immunological parameters were compared between the treatments: non‐vaccinated fish (C); bacterin‐vaccinated fish (B); bacterin plus oral booster vaccinated fish (B+O); bacterin and toxoid‐vaccinated fish (B+T) and bacterin, toxoid and oral booster‐vaccinated fish (B+T+O). Fourteen‐days vaccinated fish from B+O and B+T+O were fed with an oral booster for 4 days. After 1 week, the fish were intraperitoneally challenged with 2 × 10⁸ CFU mL^?1 of A. hydrophila. Fish from the treatment B+T+O showed the lowest cumulative mortality (11.36%) 96 h after challenge, compared with other treatments (22.72–44.04%), and a relative survival of 74%. Serum immunoglobulin in B+T+O fish was higher than in other treatments. All vaccinated fish showed an increased agglutination titre when compared with non‐vaccinated fish, both before and after challenge. Fish fed with oral booster showed an increase in phagocytic percentage before and after challenge. It can be inferred that the oral booster vaccination was efficient in reducing mortality in hybrid surubim by enhancing the response against haemorrhagic septicaemia due to A. hydrophila infection.     

Changes in the fish species composition of all Austrian lakes >50 ha during the last 150 years

Abstract  The fish communities of all Austrian natural lakes ( n  = 43) larger than 50 ha in surface area were assessed and the historical fish communities in c . 1850 were reconstructed. During the last 150 years, the fish communities of Austrian lakes have altered: in 49% of lakes at least one indigenous fish species, usually sensitive or small-bodied taxa, is now missing. Conversely, in all but one of the 43 lakes the number of fish species has increased. In particular, certain fish species of interest to angling and commercial fisheries now occur in more lakes. Generalised linear models were used to identify variables that would explain the loss of fish species using categories of lake use (ranking score). The category human population density around the lakes seemed to be one of the main causes for the loss of fish species.