光棘球海胆体腔液调理素样分子的研究

为研究光棘球海胆的体液免疫机理,首先测定在不同反应基质中等量光棘球海胆吞噬细胞对不同处理的酵母细胞的吞噬率,其次比较分析从受调理酵母细胞和非调理酵母细胞以及光棘球海胆无细胞体腔液分离出成分的样品的电泳结果,随后将光棘球海胆无细胞体腔液进行提取分离后得到调理素样分子,最后于等渗缓冲溶液中测定等量光棘球海胆吞噬细胞在不同浓度调理素样分子和不同时间点作用下的吞噬率的变化。试验结果显示,光棘球海胆吞噬细胞分别在等渗缓冲液、经酵母细胞吸附过无细胞体腔液、光棘球海胆无细胞体腔液对非调理的酵母细胞和在等渗缓冲液中对受调理的酵母细胞的吞噬率之比为1.00∶1.17∶2.00∶1.71。在3个样品里,仅有受调理酵母细胞分离出来的成分的电泳结果显示为1个明显条带。等渗缓冲溶液里,分别加入0.5、1.0、1.5 mL经纯化的调理素样分子后,4个时间点吞噬率均高于对照组,当调理素样分子浓度升高,吞噬率亦增加。试验结果表明,在光棘球海胆体腔液里含有某种调理素样分子,分子质量约156 ku,能促进光棘球海胆体腔液中吞噬细胞的吞噬作用。     

不同种海胆体腔细胞类型及体液中的酶活力

以中国主要经济海胆马粪海胆(Hemicentrotus Pulcherrimus)、虾夷马粪海胆(Strongylocentrotus intermedius)、光棘球海胆(Strongylocentrotus nudus)、紫海胆(Anthocidaris crassispina)以及杂交种海胆-马粪海胆(♀)×虾夷马粪海胆(♂)、虾夷马粪海胆(♀)×光棘球海胆(♂)、马粪海胆(♀)×紫海胆(♂)和紫海胆(♀)×光棘球海胆(♂)为对象,进行海胆的血细胞及体液中酶活力的研究。结果表明,海胆血细胞可分为4类,分别为色素细胞、纤毛游走细胞、变形吞噬细胞和无色球形细胞,比例分别为4.56%~10.96%、9.69%~12.18%、76.40%~85.36%和0.35%~1.03%;海胆血细胞体外有凝聚和吞噬现象。不同种海胆体液中酸性磷酸酶(ACP)、碱性磷酸酶(ALP)、超氧化物歧化酶(SOD)和溶菌酶(LSZ)的酶活力测定结果显示,紫海胆血清中各种酶的活性都较高,其中碱性磷酸酶、超氧化物歧化酶活性在6种海胆中都是最高的,分别为2.871 U/mg和182.149 U/mL。马粪海胆(♀)×虾夷马粪海胆(♂)的溶菌酶活性最高,为0.085 U/mL;虾夷马粪海胆(♀)×光棘球海胆(♂)的酸性磷酸酶活性最高,为1.070 U/mg,但差异均不显著(P>0.05)。[中国水产科学,2006,13(1):33-38]     

虾夷马粪海胆体腔液中调理素样分子的研究

为研究虾夷马粪海胆体液免疫机理,测定其体腔液吞噬细胞在不同介质中对不同处理的酵母细胞的吞噬作用。对其无细胞体腔液、从调理及非调理酵母细胞分离出的成分进行SDS-PAGE分析,同时对无细胞体腔液进行透析、SephadexG-200凝胶层析和SDS-PAGE分析,在等渗缓冲液和酵母细胞吸附的无细胞体腔液中,测定不同调理素样分子浓度和不同反应时间时吞噬率的变化。结果显示,吞噬细胞在3种介质:等渗缓冲液、酵母细胞吸附的无细胞体腔液和无细胞体腔液中对非调理酵母细胞吞噬率之比为1.00∶1.11∶1.94,在等渗溶液中对调理酵母细胞的吞噬率是非调理酵母细胞的1.61倍;从调理酵母细胞分离出的成分在SDSPAGE中显示一条调理素样分子带,分子量为250.62 ku。经透析和SephadexG-200凝胶层析后,测得纯化的调理素样分子的分子量为250.62 ku;在所测试的浓度范围内,调理素样分子浓度越大,吞噬活性越高,且5.0和10.0μg/mL组在各个测试时间均比对照组高。在等渗缓冲液中,反应30 min时,10.0和5.0μg/mL组与对照组均有极显著差异(P0.01),在酵母细胞吸附的无细胞体腔液中,反应30 min时,10.0μg/mL组与对照组有显著差异(P0.05)。结果表明,在虾夷马粪海胆体腔液中发现一种调理素样分子,分子量约为250.62 ku,一定浓度的调理素样分子能显著提高虾夷马粪海胆体腔液吞噬细胞的吞噬活性。     

鱼类体色成因及其调控技术研究进展(下)

2 鱼类体色变化的成因及调控 2.1 鱼类体色变化 鱼类的体色和斑纹是由鱼体色素细胞的多少、分布的区域、色素细胞内色素颗粒的状态以及虹彩细胞中反光体的反光能力强弱等决定的[9].     

养殖鱼类体色改良研究进展

鱼类在不同的生态及生理条件下会呈现不同的颜色,并且在自然界中都会呈现自己特定的体色。鱼体真皮层中含有4种色素细胞：黑色素细胞、黄色素细胞、红色素细胞及鸟粪素细胞。前3种色素细胞内的色素颗粒在神经及体液调节下,产生位移或数量的增减,及色素细胞数量的变化,加上鸟粪素细胞的特殊反光作用,使鱼体呈现不同的颜色。     

不同体色血鹦鹉鱼的色素细胞种类、数量及色素含量

用显微镜观察了体长7.0~10.0cm的黑、黑黄、黄、红4种体色血鹦鹉鱼色素细胞的形态结构,用紫外分光光度计法测定了其总类胡萝卜素含量。结果显示,(1)黑色血鹦鹉鱼通体分布树突状和圆点状两种形态的黑色素细胞,内含大量黑色素颗粒,黄色素细胞较少,颜色较浅;(2)黑黄色血鹦鹉鱼体上的树突状黑色素细胞的中心胞体扩散,分枝细长,黑色素颗粒为棕色,聚集能力弱,黄色素细胞大颜色深且数量较多;(3)黄色血鹦鹉鱼体上的黑色素细胞数量很少,黄色素细胞遍体分布,颜色深,有少量圆点状的红色素细胞,形状小,颜色浅;(4)红色血鹦鹉鱼体的黑色素细胞稀少,有少量的黄色素细胞,红色素细胞遍体分布。(5)红色血鹦鹉鱼的肉、鳞片、鳍条和皮肤的类胡萝卜素含量与其他3种体色的鱼相比差异均极显著(P0.01);黑、黑黄、黄3种体色的血鹦鹉鱼的上述4个部位的类胡萝卜素含量差异均不显著(P0.05)。本试验结果为深入探究色素细胞的发育机制提供了参考。     

南方紫海胆人工育苗试验

南方天然紫海胆经过暂养后，注射KCl溶液催产，受精卵在25．4℃海水中受精、发育，经过60天后海阳莆长到5mm以上．共剥离稚海胆30万枚，再经过90天左右培育，获得2cm左右的海胆苗16万。     

虾蟹类动物色素细胞的研究进展

综述了虾蟹类动物体色形成的色素细胞种类和调控机制。虾蟹类动物体色主要由色素细胞构成,具有躲避天敌等重要功能。虾蟹类动物的体色具有一定的遗传能力,与虾蟹类动物特有的甲壳蓝蛋白基因有直接联系。甲壳蓝蛋白主要分布在外骨骼中,与虾青素结合形成不同类型的色素细胞。虾蟹类动物的载色体转运色素颗粒,受激素调控,可通过细胞内外钙离子的运动以及cGMP信号的级联来诱导,两类肌球蛋白参与了色素转运。     

硬骨鱼类黄色素细胞的研究进展

色素细胞是动物体内产生色素的一类特化细胞,其色素颗粒可选择性吸收或反射特定波长的光产生多种颜色,从而形成动物的体色。色素细胞由胚胎神经嵴细胞分化形成,在动物警戒求偶、伪装、拟态、隐蔽以及环境适应性等一系列生物学过程中起着关键作用。黄色素细胞是仅次于黑色素细胞最受关注的研究对象。黄色素细胞合成含有类胡萝卜素和蝶啶成分的红/黄色素小体,可以形成从白色到红色等一系列的颜色,是研究色素细胞互作、色素沉着以及生物进化机制的理想模型。本综述介绍了鱼类色素细胞的来源和分布、黄色素细胞的发育和分化以及蝶啶色素合成代谢机制三个方向,并提出了现阶段黄色素细胞研究中存在的缺陷,为鱼类体色形成、色素细胞分化及体色性状分子选育提供参考依据。     

不同颜色品系暹罗斗鱼色素细胞的观察

为了解暹罗斗鱼多彩体色的成因,为斗鱼的体色选育提供科学依据,对红色系、蓝色系和金属色系暹罗斗鱼背部鳞片的色素细胞进行了研究。通过显微观察,确定了组成斗鱼鳞片的色素细胞有黑色素细胞、红色素细胞、黄色素细胞、鸟粪素细胞等4种,分布于鳞片的上下两层。研究结果表明,不同颜色品系间鳞片色素细胞的组成、分布、形状不同,是形成暹罗斗鱼多彩体色的重要原因。红色系斗鱼没有鸟粪素细胞,而蓝色系和金属色系斗鱼均有鸟粪素细胞,只分布于鳞片的下层。除了蓝色系铁锈蓝色斗鱼外,蓝色系其它颜色斗鱼和金属色斗鱼鳞片的上下层均有黑色素细胞、红色素细胞、黄色素细胞这3种色素细胞。红色系不同颜色斗鱼的色素细胞组成不同,超红色斗鱼有黑色素细胞、红色素细胞,柬红色斗鱼只有红色素细胞,橘红色斗鱼有红色素细胞和黄色素细胞,黄色斗鱼有黄色素细胞和黑色素细胞,分布于鳞片的上下层。     

Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean

The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna.     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China.     

Growth performance,digestive enzyme activity and immune response of Japanese sea bass,Lateolabrax japonicus fed with fructooligosaccharide

In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL^?1 (1.25 ng well^?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg^?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality).     

Involvement of gonadal steroids in final oocyte maturation of white perch (Morone americana) and white bass (M. chrysops): in vivo and in vitro studies

Plasma estradiol-17 (E₂), testosterone (T), 17,20-dihydroxy-4-pregnen-3-one (DHP) and 17,20,21-tri-hydroxy-4-pregnen-3-one (20-S) levels were measured by radioimmunoassay (RIA) in white perch (Morone americana) and white bass (M. chrysops) that were induced to undergo final oocyte maturation (FOM) with human chorionic gonadotropin (hCG). Plasma DHP levels increased in females of both species in association with oocyte germinal vesicle migration (GVM) and germinal vesicle breakdown (GVBD) and decreased thereafter. Plasma 20-S levels also increased with oocyte GVM in white bass, but were several-fold lower than DHP levels. Circulating E₂ and T levels were greatest during GVM and GVBD in both species and decreased to low levels during oocyte hydration and ovulation. Follicles from white perch and white bass which received a priming injection of hCG in vivo, produced both DHP and 20-S in vitro after exposure to hCG and their oocytes underwent GVBD. Ovarian incubates from unprimed fish of either species produced only E₂ and T and their oocytes did not complete GVBD. Oocytes from unprimed bass, but not perch, matured when follicles were exposed to hCG in vitro. Both trilostane and cycloheximide blocked in vitro production of DHP and 20-S and oocyte GVBD by white perch follices. DHP and 20-S were equipotent inducers of FOM in the GVBD bioassay. None of several other structurally-related steroids tested were effective within a physiological range of concentrations. These results indicate a role for DHP and 20-S in the control of FOM in white perch and white bass.     

Cardiac,ventilatory and metabolic responses of two ecologically dissimilar species of fish to waterborne cyanide

Changes in heart rate, ventilatory activity and oxygen consumption were determined in trout (Salmo gairdneri) and brown bullhead catfish (Ictalurus nebulosus) during exposure to a steadily increasing concentration of waterborne cyanide selected to produce death in 8–9 hours for each species. The lethal cyanide concentration for the bullheads was an order of magnitude higher than for trout. Trout developed an immediate and gradually increasing bradycardia throughout the exposure period. Cyanide produced tachycardia in the bullhead followed by a gradual onset of bradycardia as the concentration of cyanide was raised. Pericardial injection of atropine (a muscarinic cholinergic antagonist) indicated that bradycardia in the trout was due initially to increased vagal tone but later due to the direct effect of cyanide on the heart. Hyperventilation in the trout persisted throughout the exposure period, although the rate and amplitude fluctuated and was variable between individual fish. During the last hour of exposure (highest cyanide concentration), ventilation was characterized by rapid, shallow breaths followed by a sudden respiratory arrest. The bullheads exhibited hyperventilation during the first 3 hours of exposure followed by a gradual, linear drop in ventilation rate and amplitude until death occurred. Cardiac and ventilatory responses in both species were attributed to stimulation of central and peripheral chemoreceptors by cyanide. Evidence is presented which suggests the initial response in the bullheads was due, at least in part, to gustatory stimulation by the cyanide. Oxygen consumption of the trout remained above pre-exposure levels for the majority of the test period. Oxygen consumption in the bullhead paralleled the changes in heart and ventilatory rates. Whole-body lactate levels of fingerlings of both species during cyanide exposure were measured to estimate the extent of anaerobiosis. Whole-body lactate levels were much greater in the bullheads than the trout, indicating a higher capacity for anaerobiosis, possibly due to a greater fuel supply. Overall, the trout responded to cyanide in a manner similar to that produced by environmental hypoxia whereas the bullheads experienced a gustatory stimulus which masked the hypoxia-like response.     

Protein and amino acid composition from the mantle of juvenile O ctopus vulgaris exposed to prolonged starvation

Protein and amino acid composition of the mantle of juvenile O ctopus vulgaris (Cuvier, 1797) during fasting for 27 days were determined. Average protein content of octopus mantle was of 711.19 ± 46.80 g kg^?1 DW, and it decreased with increasing fasting days. The non‐essential amino acids content was higher (486.18 ± 11.08 g kg^?1 protein) than essential amino acids (425.82 ± 9.15 g kg^?1 protein) at the start of the experiment (unstarved animals). The results suggest that the amino acid profile of the mantle where the most abundant amino acids are Arg, His, Lys, Gly, Leu and Pro could indicate a prolonged fasting condition (>20 days) or poor nutrition of O . vulgaris. This study supports the idea of using mantle for metabolic needs of starved O . vulgaris suggesting that the degradation pathway of amino acids to pyruvate and tricarboxylic acid cycle intermediates was favoured contrary to the degradation pathway of ketogenic amino acids. Special considerations should be taken concerning Thr, Ile, Ser, Ala, Asx (Asp, Asn), Glx (Glu, Gln) (because of their fast intake) and Lys and His (due to their stable contents) during a prolonged period of fasting.     

An overview of stress physiology of fish transport: changes in water quality as a function of transport duration

This study brings an integrated analysis about the relationship between water deterioration and its physiological consequences in live fish transport. The analysis was focused on the transport water and its deterioration, and physiological challenges imposed on the fish. Usual commercial handling procedures employed to mitigate fish stress during transport were discussed. Future topics of research for the establishment of safer fish transport protocols were proposed. Transport was classified into short (≤8 h) or long transport (>8 h). The main issue in short transports should be the prevention of water pH reduction, while in long transports it is the increase in ammonia. Plasma cortisol is the most employed marker for stress and is acutely elevated upon short episodes of transport, but remains elevated even in long‐transport events. Plasma glucose is perhaps a better marker for handling stress. Plasma lactate, pH, osmolality CO₂ and ions should be more often evaluated. Plasma Na⁺ and Cl^‐ are very useful markers of acidosis, due to their respective exchange for H⁺ and , for acid–base regulation. The establishment of species‐specific transport protocols should be preceded by such combined analyses of water and physiological parameters.     

Are hatchery‐reared abalone naïve of predators? Comparing the behaviours of wild and hatchery‐reared northern abalone,Haliotis kamtschatkana (Jonas, 1845)

Abalone populations have declined worldwide, generating interest in enhancement using hatchery‐reared individuals. In many cases, such restoration efforts have met with limited success due to high predator‐induced mortality rates. Furthermore, the mortality rates of outplanted hatchery abalone are often considerably higher than for wild individuals. This study uses northern abalone (Haliotis kamtschatkana) as a case study to determine whether hatchery‐reared abalone behave differently than their wild counterparts. In the field, outplanted hatchery‐reared abalone were significantly less responsive than wild abalone, in terms of number of abalone responding and intensity of response, to nearby movement and to physical contact with an inert probe. Also, when encountering a cue to which all abalone responded (a seastar predator), hatchery‐reared individuals remained subdued. Anti‐predator behavioural deficits in hatchery‐reared abalone were more pronounced in 4‐year‐old individuals than in 1‐year‐old individuals, suggesting an influence of either age or amount of time spent in the hatchery environment. These behavioural differences are expected to increase the vulnerability of hatchery‐reared abalone to predators, and are likely a major cause of their elevated predator‐induced mortality when outplanted.