金乌贼同工酶分析

采用水平淀粉凝胶电泳技术对金乌贼（Sepia esculenta）同工酶的组织特异性及其山东日照近海群体的遗传结构进行研究。对金乌贼眼、鳃、肌肉、口球、肝脏、鳃心6种组织的19种同工酶进行分析，检测出PGDH、GPI、MPI、IDHP、SOD、ME、AAT、DIA、MDH、LDH、G3PDH、PGM共12种同工酶在几种组织中有较稳定而清晰的表达。结果表明，金乌贼同工酶的表达有明显的组织特异性。选择金乌贼3种组织（眼、口球、鳃心）进行同工酶分析，共记录了18个基因座位，其中3个基因座位LDH-2^＊、G3PDH-1^＊、PGM^＊呈多态，其多态座位比例为0．1667（P0．99）和0．0556（P0.95），平均观测杂合度和预期杂合度分别为0．0159和0．0143，平均有效等位基因数为1．0201，表明日照近海金乌贼群体的遗传多样性较低。     

两种壳色虾夷扇贝的同工酶分析

采用不连续聚丙烯酰胺凝胶垂直电泳技术对两种壳色虾夷扇贝的闭壳肌进行了同工酶检测分析与比较。8种同工酶（MDH、ADH、ME、SOD、EST、GDH、SDH和α-AMY）共检测出17个位点,其中白色贝有7个多态位点,褐色贝有6个多态位点,多态位点（P0.99）的比例分别为41．18％和35．29％,白色贝和褐色贝平均每个位点的等位基因有效数目（Ac）分别为1．1421和1．1040,预期杂合度分别（Hc）为0．0919和0．0674,实际杂合度（Ho）分别为0．1275和0．0907,多态位点的遗传偏离指数表明,白色贝具有更高的遗传变异水平和多样性水平。住点Est-3可以作为鉴定白色贝和褐色贝的特异性蛋白质分子标记。     

红鳍东方鲀6种同工酶的组织特异性及基因位点分析

采用水平淀粉凝胶和聚丙烯酰胺凝胶垂直电泳法,对2龄野生红鳍东方鲀的肌肉、肝脏、肾脏、心脏、眼、鳃6种组织中的苹果酸酶(ME),乳酸脱氢酶(LDH),异柠檬酸脱氢酶(IDH),葡萄糖脱氢酶(GLCDH),磷酸葡萄糖变位酶(PGM)和酯酶(EST)共6种同工酶进行了研究。结果表明,以上6种同工酶存在明显的组织特异性。LDH在除肝脏外的5种组织中活性很强,GLCDH只在肝脏中表达,鳃组织中的GLCDH和肌肉组织中的EST表达活性都很弱。6种同工酶共检测到12个基因位点,其中ME和PGM有两个基因位点编码,EST有5个基因位点编码。本试验还检测到Ldh-1、Est-1、Est-2、Est-3和Est-4共5个基因位点具有多态现象(P0·99),多态位点比例为41·67%,在12个基因位点共观察到17个等位基因表达,平均每个位点表达的等位基因数为1·417个。同时在红鳍东方鲀眼组织中发现,AB3型和B4型LDH间有一条酶带弱表达。     

红鳍东方纯养殖群体生化遗传变异初步研究

采用水平凝胶电泳技术对红鳍东方纯养殖群体进行了同工酶分析.本实验采用肌肉和肝脏两种组织,分析了8种酶,检测到9个位点,在0.99水平上,PGM*和IDHP*表现为多态,MPI*、LDH*、AAT*、SOD*、MDH-1*、MDH-2*和G3PDH*均为单态,多态位点比例为0.222 2,平均预期杂合度为0.099 8,实际观测杂合度为0.046 7,平均有效等位基因数目为1.184 0,PGM*位点的遗传偏离指数为-0.070 7,杂合子缺失,IDHP*位点无杂合子,遗传偏离指数为-1.     

红鳍东方鲀养殖群体生化遗传变异初步研究

采用水平凝胶电泳技术对红鳍东方鲀养殖群体进行了同工酶分析。本实验采用肌肉和肝脏两种组织,分析了8种酶,检测到9个位点,在0.99水平上,PGM*和IDHP*表现为多态,MPI*、LDH*、AAT*、SOD*、MDH-1*、MDH-2*和G3PDH*均为单态,多态位点比例为0.2222,平均预期杂合度为0.0998,实际观测杂合度为0.0467,平均有效等位基因数目为1.1840,PGM*位点的遗传偏离指数为-0.0707,杂合子缺失,IDHP*位点无杂合子,遗传偏离指数为-1。     

黄、渤海半滑舌鳎种群遗传结构的同工酶分析

采用水平淀粉凝胶电泳技术检测黄海、渤海半滑舌鳎两个群体的同工酶组织特异性并分析和研究其群体遗传结构。结果表明,半滑舌鳎的眼、鳃、肌肉、心脏、肝脏、肾脏、脾脏、鳍条、脑等9种组织和器官对18种同工酶的表达呈现明显的组织特异性。对肌肉和肝脏两种组织的PGDH、MPI、IDHP、SOD、AAT、MDH、LDH、G3PDH、PGM、LAP、ADH、SDH和ALP共13种同工酶进行了遗传学分析,共记录16个基因座位。其中,黄海群体在4个基因位点(IDHP、PGM、G3PDH、SDH)、渤海群体在5个基因位点(IDHP、PGM、MDH2、SDH、ALP)表现为多态,黄海和渤海群体的多态位点比例分别为0.2500和0.3125;两个群体的平均观测杂合度分别为0.0143、0.0104,平均预期杂合度分别为0.0140、0.0102;平均有效等位基因数则分别为1.0151和1.0108。两群体间的遗传距离为0.00012。半滑舌鳎的两个群体遗传多样性水平较低,两群体间无明显的遗传分化。     

中国沿海5个自然西施舌群体等位酶的遗传变异研究

采用聚丙烯酰胺凝胶垂直平板电泳技术,研究了中国沿海的山东胶南(JN)、江苏南通(NT)、浙江台州(TZ)、福建福州(FZ)、广西北海(BH)西施舌(Coelomactra antiquata)自然群体的等位基因酶遗传变异,对其斧足、外套膜和肝脏组织进行了13种同工酶的比较实验,最后选择肝脏作为西施舌的等位酶研究组织.选取谱带清晰的8种等位酶进行检测,共检测到17个位点.JN、NT、TZ、FZ、BH群体的多态位点比例分别为58.82%、70 59%、50.00%、56.25%、41.17%,平均有效等位基因数(Ae)分别为1.550、1.555、1.434、1.602、1.828,平均杂合度观察值(Ho)分别为0.240、0.270、0.239、0.243、0.288.结果表明,群体的遗传变异水平BH>NT>FZ>JN>TZ,中国沿海西施舌遗传变异水平较高、种质资源较好;同时发现除浙江台州(TZ)群体外,普遍存在杂合子缺失现象;并根据等位酶资料对5个群体进行了UPGMA聚类分析.     

中国沿海5个自然西施舌群体等位酶的遗传变异研究

采用聚丙烯酰胺凝胶垂直平板电泳技术,研究了中国沿海的山东胶南(JN)、江苏南通(NT)、浙江台州(TZ)、福建福州(FZ)、广西北海(BH)西施舌(Coelomactra antiquata)自然群体的等位基因酶遗传变异,对其斧足、外套膜和肝脏组织进行了13种同工酶的比较实验,最后选择肝脏作为西施舌的等位酶研究组织。选取谱带清晰的8种等位酶进行检测,共检测到17个位点。JN、NT、TZ、FZ、BH群体的多态位点比例分别为58.82%、70.59%、50.00%、56.25%、41.17%,平均有效等位基因数(Ae)分别为1.550、1.555、1.434、1.602、1.828,平均杂合度观察值(Ho)分别为0.240、0.270、0.239、0.243、0.288。结果表明,群体的遗传变异水平BH>NT>FZ>JN>TZ,中国沿海西施舌遗传变异水平较高、种质资源较好;同时发现除浙江台州(TZ)群体外,普遍存在杂合子缺失现象;并根据等位酶资料对5个群体进行了UPGMA聚类分析。     

蓝色鳞鲤(Cyprinus carpio blue var)同工酶分析

采用聚丙烯酰胺凝胶垂直板电泳法对蓝色鳞鲤(Cyprinus carpio blue var)进行同工酶分析。结果表明:在蓝色鳞鲤的眼睛、肌肉、肝脏、心脏、肾脏5种组织中,14种酶(LDH、ADH、GDH、MDH、G-6-PDH、EST、POD、SDH、FDH、SOD、α-AMY、CAT、COX、ME)的同工酶谱均存在明显的组织特异性。14种酶共记录出33个基因位点,其中α-Amy-2、Cox-2、Est-1、Ldh-1、Mdh-1、Mdh-2和Sod-1为多态位点。蓝色鳞鲤群体的多态位点百分数为21.21%(P0.99),平均预期杂合度和平均实际杂合度分别为0.1079和0.2121,遗传偏离指数(d)值为正。平均有效等位基因数(Ne)为1.24。实验表明,目前蓝色鳞鲤群体的种质资源状况尚好,表现出明显的杂交优势。     

日本蟳同工酶组织特异性及生化遗传分析

以日本虫寻为研究材料,采用聚丙烯酰胺凝胶垂直平板电泳方法,分析了肌肉、眼球、鳃、心脏和肝胰腺5种器官或组织中LDH、ADH、MDH、ACP、AKP、GDH、R-AMY、α-AMY、MEP、EST和SOD共11种同工酶的表达情况。日本虫寻体内同工酶的表达具有明显的组织特异性。在实验中共检测了38个位点,其中Adh、Est-3、M ep-5、α-Amy-1、sMdh-3和Akp-4共6个多态位点,多态位点比例为15.79%,平均杂合度为0.09。     

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.     

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).     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.     

Effects of cortisol on hepatic carbohydrate metabolism and responsiveness to hormones in the sea raven,Hemitripterus americanus

The sea raven, Hemitripterus americanus, is a sit-and-wait, low metabolic rate, marine teleost. The objective of this study was to determine i) whether cortisol implantation (50 mg. kg^-1) for 7 days altered hepatocyte metabolism, and hepatocyte responsiveness to epinephrine, glucagon and insulin, and ii) whether 8 weeks of food-deprivation modified the above response. Cortisol implantation significantly increased hepatocyte total glucose production and oxidation from alanine compared to the sham group. There was no cortisol effect on glycogen breakdown, suggesting that the activation of other pathways, including gluconeogenesis, are required to account for the increased glucose production. Epinephrine-mediated (10^-5M) glycogen breakdown and insulin-mediated (10^-8M) total glucose production were enhanced in hepatocytes of cortisol implanted sea ravens, but there were no change in any glucagon (10^-7M) effects. The enhanced glycogen breakdown in the absence of similar increases in total glucose production with epinephrine indicates mobilization of carbohydrate reserves for endogenous use by the liver. Food-deprivation for 8 weeks significantly decreased condition factor, plasma cortisol concentration and liver glycogen content in the sea raven, but had no effect on plasma glucose concentration. Hepatocyte total glucose production and flux rates from alanine increased significantly with food-deprivation. Moreover, food-deprivation increased responsiveness of total hepatocyte glucose production to the actions of glucagon and insulin, but not to epinephrine; none of these effects were modified by cortisol implantation. Our results indicate that cortisol in the sea raven exerts both a direct and an indirect or permissive effect on hepatocyte metabolism by modifying hepatocyte responsiveness to epinephrine and insulin stimulation. Cortisol implantation did not modify the effects of glucagon or food-deprivation in this species.     

Simulation and quantification of enrichment and retention processes in the southern Benguela upwelling ecosystem

Important environmental processes for the survival and recruitment of early life stages of pelagic fishes have been synthesized through Bakun's fundamental triad as enrichment, concentration and retention processes (A. Bakun, 1996, Patterns in the Ocean. Ocean Processes and Marine Population Dynamics. San Diego, CA, USA: University of California Sea Grant). This conceptual framework states that from favourable spawning habitats, eggs and larvae would be transported to and/or retained in places where food originating from enrichment areas would be concentrated. We propose a method for quantifying two of the triad processes, enrichment and retention, based on the Lagrangian tracking of particles transported within water velocity fields generated by a three‐dimensional hydrodynamic model. We apply this method to the southern Benguela upwelling ecosystem, constructing putative maps of enrichment and retention. We comment on these maps regarding main features of the circulation in the region, and investigate seasonal variability of the processes. We finally discuss the results in relation to available knowledge on the reproductive strategies of two pelagic clupeoid species abundant in the southern Benguela, anchovy (Engraulis encrasicolus) and sardine (Sardinops sagax). Our approach is intended to be sufficiently generic so as to allow its application to other upwelling systems.     

Effects of capture,loading density and transport stress on the mortality,physiological responses,bacterial density and growth of rohu <Emphasis Type="Italic">Labeo</Emphasis><Emphasis Type="Italic"> rohita</Emphasis> fingerlings

This study evaluated the effects of three levels of loading density (200, 300 and 400 g L⁻¹) and four levels of duration (1, 3, 6 and 9 h) on the mortality, plasma cortisol and chloride levels and growth of rohu Labeo rohita fingerlings in a transport simulation. While total immediate mortality was low (4–12%), delayed mortality was high (27–49%). Initial capture and handling were found to constitute the most stressful phase as indicated by elevated plasma cortisol. The levels of cortisol continued to rise at 6 and 9 h after transport, with the highest level observed at 9 h after transport. The level of plasma chloride fell after transport across all density levels and treatment durations. Specific growth rate (SGR, %/day) and total bacterial density (cfu/g) were not different between and within treatments, and between treatments and the controls. This study demonstrates that transport has a profound effect on the stress responses of rohu fingerlings. Although the level of immediate mortality was low, the higher level of delayed mortality has serious implications in terms of production inefficiency for grow-out farmers.     

The effects of introduced tilapias on native biodiversity

• 1. The common name ‘tilapia’ refers to a group of tropical freshwater fish in the family Cichlidae (Oreochromis, Tilapia, and Sarotherodon spp.) that are indigenous to Africa and the southwestern Middle East. Since the 1930s, tilapias have been intentionally dispersed worldwide for the biological control of aquatic weeds and insects, as baitfish for certain capture fisheries, for aquaria, and as a food fish. They have most recently been promoted as an important source of protein that could provide food security for developing countries without the environmental problems associated with terrestrial agriculture. In addition, market demand for tilapia in developed countries such as the United States is growing rapidly.
• 2. Tilapias are well‐suited to aquaculture because they are highly prolific and tolerant to a range of environmental conditions. They have come to be known as the ‘aquatic chicken’ because of their potential as an affordable, high‐yield source of protein that can be easily raised in a range of environments — from subsistence or ‘backyard’ units to intensive fish hatcheries. In some countries, particularly in Asia, nearly all of the introduced tilapias produced are consumed domestically; tilapias have contributed to basic food security for such societies.
• 3. This review indicates that tilapia species are highly invasive and exist under feral conditions in every nation in which they have been cultured or introduced. Thus, the authors have concluded that, despite potential or observed benefits to human society, tilapia aquaculture and open‐water introductions cannot continue unchecked without further exacerbating damage to native fish species and biodiversity. Recommendations include restricting tilapia culture to carefully managed, contained ponds, although exclusion is preferred when it is feasible. Research into culture of indigenous species is also recommended.

Copyright © 2005 John Wiley & Sons, Ltd.