牙鲆头肾巨噬细胞的分离培养与鉴定

本研究使用密度梯度离心法从牙鲆(Paralichthys olivaceus)头肾中分离得到了巨噬细胞,通过差速贴壁法对获得的细胞进行纯化,后续经过优化培养条件和培养过程,采用L-15培养基(Gibco)、5％胎牛血清、1％青–链霉素、1％非必需氨基酸、30％L929细胞培养基在24℃、无CO2的条件下进行培养.显微镜...     

Salmonid colonization of new streams in Glacier Bay National Park, Alaska

Abstract. Following the rapid recession of a neoglacial ice sheet within the last 250 years, colonization of recently deglaciated streams by salmonid fishes was investigated in Glacier Bay National Park, south-eastern Alaska. The primary factors governing the establishment, species diversity composition and abundance of salmonids in Glacier Bay streams were water temperature, sediment loading and stream discharge. No salmonids were found in the turbid meltwater streams emerging from retreating ice. Coho, Oncorhynchus kisutch (Walbaum), and sockeye, Oncorhynchus nerka (Walbaum). salmon and Dolly Vardcn, Salvelinus malma (Walbaum), charr were the first salmonids to colonize the youngest clearwater stream. Juvenile Dolly Varden were more abundant than juvenile coho salmon in the most recently formed clearwater stream because of the characteristic absence of pool habital. Densities of juvenile coho salmon were six times greater in a stream with a series of lakes compared with a stream of similar age without lakes. Future advancement of salmonid stocks will probably depend upon the rate and extent of the development of riparian vegetation and inputs of large woody debris from the developing forest to provide further instream cover, habitat variation and channel stabilization.     

Salmonid whirling disease: myxosporean and actinosporean stages cross-react in direct fluorescent antibody test

Abstract. Serologic relatedness of the two life stages of the salmonid whirling disease parasite Myxosoma cerebralis Hofer, 1903 — myxosporean spores from fish cartilage and actinosporean triactinomyxon spores from aquatic tubificids — were investigated. When the direct fluorescent antibody technique was used, anti-triactinomyxon and anti- M. cerebralis rabbit sera conjugated with fluorescein isothiocyanate cross-reacted with the respective heterologous life stage. Both stages showed similar locations of specific fluorescence with conjugates of either homologous or heterologous serum. Thus, serology supports the relatedness of the myxosporean M. cerebralis and the actinosporean triactinomyxon stages.     

Isolation, cultivation and characterization of head kidney macrophages from Atlantic cod, Gadus morhua L.

Head kidney macrophages from Atlantic cod, Gadus morhua L., were isolated by density sedimentation and maintained under serum-free conditions for up to one week. The cells adhered and spread well on glass and plastic, were highly phagocytic, and had typical macrophage morphology as shown by phase contrast and scanning and transmission electron microscopy. Histochemical studies with the light microscope showed that the macrophages were acid phosphatase and non-specific esterase positive, and alkaline phosphatase and peroxidase negative. Compared with unstimulated control cells, LPS-treated cells showed enhanced superoxide anion formation, as measured by the reduction of nitroblue tetrazolium, and increased levels of acid phosphatase activity.     

A Review of Di-n-Butylphthalate in the Aquatic Environment: Concerns Regarding Its Use in Salmonid Aquaculture

The presence and effects of di-n-butylphthalate (DBP) in aquatic environments are reviewed. Particular attention is paid to the use of DBP as a solvent in the pesticide formulation, Aquagard®, used to treat salmonids against external parasites at aquaculture sites in Europe. DBP is sparingly soluble in water, is readily metabolized by fish, and has a high lethal threshold for aquatic organisms. It is, however, lipophilic, persistent in aquatic sediments, and is listed as a priority pollutant by the governments of Canada and United States. The dissemination of this compound directly to water raises several concerns which should be addressed by further research and increased monitoring.     

Optimising of culture conditions and stimulation of head kidney macrophages from Atlantic cod, Gadus morhua L.

The cultivation of Atlantic cod, Gadus morhua L., head kidney macrophages was optimal at 4–6 °C. The viability of cells in culture was examined by the trypan blue exclusion test 2, 5 and 7 days after seeding. Simultaneously, the number of adherent cells at the various times was checked. The morphology of the cells was also evaluated during the cultivation period. Enhanced respiratory burst activities were demonstrated in macrophages after stimulation with known immunostimulants like lipopolysaccharide, tuftsin and an acid peptide fraction previously shown to stimulate Atlantic salmon leucocytes efficiently. Generally, the Atlantic cod macrophages seemed to respond to a lesser extent compared with macrophages obtained from both mammalian and other fish species.     

Performance of a closed recirculating system with foam separation, nitrification and denitrification units for intensive culture of eel: towards zero emission

The development of a closed recirculating aquaculture system that does not discharge effluents would reduce a large amount of pollutant load on aquatic bodies. In this study, eel were reared in a closed recirculating system, which consisted of a rearing tank, a foam separation unit, a nitrification unit and a denitrification unit. The foam separation unit has an inhalation-type aerator and supplies air bubbles to the rearing water. The growth of eel, which were fed a commercial diet, was satisfactory, with gross weight increases of up three times in 3 months. The survival rate under the congested experimental conditions was 91%. The foam separation unit maintained oxygen saturation in the rearing water at about 80%. Furthermore, fine colloidal substances were absorbed on the stable foam formed from eel mucus and were removed from the rearing water by foam separation. Ammonia oxidation and the removal of suspended solids were accomplished rapidly and simultaneously in the nitrification unit. The ammonia concentration and turbidity were kept at less than 1.2 mg of N per litre and 2.5 units, respectively. When the denitrification process was operated, nitrate that accumulated in the rearing water (151 mg of N per litre) was reduced to 40 mg of N per litre. The sludge was easily recovered from the nitrification and denitrification tanks, and the components were found suitable as compost. Based on these results, the intensive aquaculture of freshwater fish such as eel can be achieved using a closed recirculating system without emission.     

Barnacle culture: background,potential and challenges

There are approximately a dozen species of commercially interesting barnacles worldwide, some of which have been cultured on a semi‐industrial scale. These species are listed and information is provided with regard to geographical distribution, landings and prices. Traditionally, ‘goose’ barnacles (four species) are considered to be the most important for consumption. World production already stands at 500 tonnes year^?1, but this species has not been cultured to date. Some ‘acorn’ barnacles are also consumed (seven species), with harvest levels per species that do not exceed 200 tonnes year^?1 and selling prices that can reach US$17/kg. ‘Acorn’ barnacle culture on a world scale is still developing. Nevertheless, production has occurred on a semi‐industrial scale; specifically, spat have been collected from the wild and grown in suspended systems. Farming trials have focused on two species of acorn barnacles: Austromegabalanus psittacus (Molina 1782) ‘picoroco’ in Chile and Megabalanus azoricus (Pilsbry 1916) ‘craca’ in Portugal. The large‐scale production of these crustaceans will depend on the optimization of spat collection from the wild and/or the parallel development of mass production technologies for larvae (hatcheries). In addition, further development will be achieved by opening up new markets for commercialization.     

In vivo and in vitro assessment of Lactobacillus acidophilus as probiotic for tilapia (Oreochromis niloticus,Perciformes:Cichlidae) culture improvement

In this study, the use of Lactobacillus acidophilus as a probiotic for Nile tilapia (Oreochromis niloticus) culture was studied. Fish survival and the expression of some genes involved in the immune response were assessed. Diet supplementation with L. acidophilus for 15 days caused a significant increase in fish survival during a challenge with Aeromonas hydrophila and variations in immune response related to IL‐1β and transferrin expression in Nile tilapia spleen and kidney. Moreover, extracellular products (ECPs) of L. acidophilus showed high antibacterial activity against fish pathogens such as A. hydrophila and Streptococcus agalactiae in vitro. It was also observed that viable L. acidophilus was able to disrupt quorum sensing activity in Chromobacterium violaceum.     

黄芪对鲤头肾中巨噬细胞的增殖和一氧化氮产量的影响：离体研究

用密度梯度离心分离鲤头肾中的巨噬细胞和嗜中性粒细胞。用RPMI细胞培养液对分离的细胞进行原代培养,在细胞培养液中加入不同浓度的黄芪水提取液来研究黄芪对鲤头肾中免疫细胞非特异性免疫应答的影响。黄芪水提取液单独使用时能刺激鲤头肾中巨噬细胞的增殖,但不能刺激巨噬细胞的氮暴发活性。用黄芪和脂多糖(LPS)混合刺激巨噬细胞和中性粒细胞时,黄芪水提取液能显著提高脂多糖刺激所产生的一氧化氮的产量。研究结果表明,黄芪不仅能刺激巨噬细胞数量的增加,而且能协同脂多糖增强头肾中免疫细胞的功能,从而对机体的非特异性免疫起调节作用。     

Great scallop, Pecten maximus, research and culture strategies in Norway: a review

The great scallop, Pecten maximus is a potentialaquaculture species in Norway, but production to date has been low. Greatscallop landings in 2000 were 571 tonnes, exclusively from harvesting wildstocks, landed by divers. Approximately two million juveniles of 15mm shell height have been produced annually in a hatchery since1998, and distributed to farmers. Research projects on larvae and juvenilesinclude studies of various antibacterial treatments, improved culture systemdesigns and application of probiotics. Results from hanging cultures underdifferent environmental conditions indicate a strong correlation between lowtemperature and poor survival, and point out the importance of careful selectionof cultivation sites. Good growth potential of scallops in Norwegian waters hasbeen shown, and it is possible to reach market size of 10 mm shellheight in three to four years. Experiments with fences and other strategiesprotecting cultured scallops on the seabed from predation by crabs are inprogress.     

海水池塘养殖模式优化：概念、原理与方法

随着海洋渔业资源衰退和人类社会对水产品需求的持续增长,海水养殖规模逐年扩大,养殖生产对环境的影响也日益严重[1-6]。现已确认海水鱼虾类养殖是造成海洋沿岸带水域富营养化的重要污染源之一[5-8],这一严峻现实要求今后在发展海水养殖产业时不仅需要宏观上对养殖区域和养     

Morphometric characterization of a small-sized strain of Brachionus plicatilis in culture

The evolution of size frequency distribution in populations of three strains (Bs, S-1 and UK) of Brachionus plicatilis growing in cultures fed with Nannochloris oculata has been studied. Also the size frequency distribution of Bs has been determined in a natural population sample and in the exponential growth phase of two cultures of this strain fed with Tetraselmis suecica and Nannochloris maculata, respectively. For morphometric characterization of these three rotifer strains the anterior medial spine length to body length relationship was examined.The results shows a clear difference between the Bs strain and the two other strains in frequency distribution and range of size. At the end of the exponential phase S-1 and UK strains when cultured with Nannochloris oculata, could supply only 1 or 2% of individuals smaller than 150 μm while Bs strain could supply 80%.The Bs strain covers a range below 150 μm and may be appropriate as early food in the large-scale rearing of some marine species.     

In vitro effect of levamisole on the cell viability,phagocytosis and respiratory burst of Barbel chub (Squaliobarbus curriculus) macrophages

The present study was to determine in vitro effect of levamisole on the immune functions of Barbel chub (Squaliobarbus curriculus), head–kidney‐derived (HKM), spleen‐derived (SM) and peripheral blood monocyte‐derived macrophage (PBM). Macrophages were incubated with levamisole 10³, 10¹, 10⁻¹ and 10⁻³ ng mL⁻¹ to assay the cell viability, respiratory burst and phagocytosis. The results showed that macrophages treated with levamisole 10⁻³ ng mL⁻¹ gave a maximum respiratory burst response, whereas levamisole 10³ ng mL⁻¹ had no effect. Phagocytosis activity of the macrophages treated with levamisole enhanced significantly when compared to control, maximum response being at 10⁻³ ng mL⁻¹. While using the methylthiazoletetrazolium method to measure the cell activity for 12, 24, 36, 48, 60 h, there was no significant role on proliferation and the cell viability began to decline after 24 h. In conclusion, levamisole is a potent enhancer of Barbel chub macrophage activity with low concentration.     

Gluconeogenesis in trout (Oncorhynchus mykiss) white muscle: purification and characterization of fructose-1,6-bisphosphatase activity in vitro

Studies of the enzyme fructose-1,6-bisphosphatase (FBPase) of rainbow trout (Oncorhynchus mykiss) have been undertaken in order to illuminate aspects of skeletal muscle gluconeogenesis in these animals. Maximal activities in crude homogenates of several organs suggest that the liver possesses the greatest FBPase activity on a unit g^–1 tissue basis but that the white muscle, owing to its bulk, contributes substantially to whole body FBPase activity. Studies of fructose-6-phosphate-1-kinase (PFK) and FBPase in crude homogenates of several organs suggests an important role for intracellular pH in regulating the relative carbon flux through the FBPase/PFK locus in vivo. Furthermore, a three-step purification scheme is described for trout white muscle FBPase by which a stable and homogeneous (by SDS PAGE) enzyme preparation (isoelectric point = 7.2; molecular weight = 37.6 kd) was obtained. Kinetic studies of the purified enzyme were undertaken at 20°C under conditions reflective of "rest" and "exercise/recovery" intramuscular pH in vivo. Affinity for substrate (F-1,6-P₂) was increased (Km = 6.88 versus 2.44 mol 1-^–1 as was enzyme activity when pH was lowered from 7.0 to 6.5. Various inhibitor metabolites are identified including F-2,6-P₂ (mixed-type inhibitor, K_i = 0.201 mol 1^–1, pH 7.0) and AMP (non-competitive inhibitor, K_i = 0.438 mol 1^–1, pH 7.0). Inhibition by F-2,6-P₂ was strongly alleviated by a reduction in pH from 7.0 to 6.5 (I₅₀ increased from 0.14 to 0.32 mol 1^–1). AMP on the other hand was a more potent inhibitor at pH 6.5 but this inhibition was totally reversed under conditions of citrate, NH₄ ⁺ and AMP typical of muscle during recovery from exercise in vivo. In purified white muscle enzyme preparations, FBPase demonstrated maximal activity at pH 6.5 whereas the optimal pH of PFK was 7.0 or greater. Indeed, it appears from these in vitro data that regulation by metabolite levels as well as pH are required for net FBPase flux in vivo. It is concluded, therefore that trout white muscle FBPase demonstrates the potential to play an important enzymatic role in the control of intramuscular gluconeogenesis in these animals. The results are discussed in relation to present knowledge regarding the metabolic responses of trout white muscle to, and its subsequent recovery from, exhaustive exercise.     

White tail disease of the giant freshwater prawn, Macrobrachium rosenbergii: separation of the associated virions and characterization of MrNV as a new type of nodavirus

White tail disease of the farmed freshwater prawn, Macrobrachium rosenbergii, is the cause of mortalities in the French West Indies, China and India. Two different sized particles, both developing in the cytoplasm of target cells, are found associated with diseased animals. These two viruses were separated, purified and subsequently characterized. The larger one, called MrNV, is icosahedral in shape and 27 nm in diameter. Its genome is composed of two fragments of linear single-stranded RNA (ss-RNA), of 2.9 and 1.3 kb, respectively and its capsids exhibited a single polypeptide of 43 kDa. These characteristics and the partial sequence of a cloned fragment of RNA-1 suggest this agent is a member of the family Nodaviridae, but with differences from both the genera Alphanodavirus and Betanodavirus. The smaller virus, named XSV, is icosahedral in shape, 15 nm in diameter, possesses a linear ss-RNA genome of about 0.9 kb, and its capsid exhibits two polypeptides of 16 and 17 kDa, respectively. The relationships between these two viruses remain unknown.