High yield and rapid growth of Neoparamoeba pemaquidensis in co-culture with a rainbow trout gill-derived cell line RTgill-W1

Neoparamoeba pemaquidensis is an ubiquitous amphizoic marine protozoan and has been implicated as the causative agent for several diseases in marine organisms, most notably amoebic gill disease (AGD) in Atlantic salmon. Despite several reports on the pathology of AGD, relatively little is known about the protozoan and its relationship to host cells. In this study, an in vitro approach using monolayers of a rainbow trout gill cell line (RTgill-W1, ATCC CRL-2523) was used to rapidly grow large numbers of N. pemaquidensis (ATCC 50172) and investigate cell-pathogen interactions. Established cell lines derived from other tissues of rainbow trout and other fish species were also evaluated for amoeba growth support. The amoebae showed preference and highest yield when grown with RTgill-W1 over nine other tested fish cell lines. Amoeba yields could reach as high as 5 x 10(5) cells mL(-1) within 3 days of growth on the gill cell monolayers. The amoebae caused visible focal lesions in RTgill-W1 monolayers within 24 h of exposure and rapidly proliferated and spread with cytopathic effects destroying the neighbouring pavement-like cells within 48-72 h after initial exposure in media above 700 mOsm kg(-1). Disruption of the integrity of the gill cell monolayers could be noted within 30 min of exposure to the amoeba suspensions by changes in transepithelial resistance (TER) compared with control cell monolayers maintained in the exposure media. This was significantly different by 2 h (P < 0.05) compared with control cells and remained significantly different (P < 0.01) for the remaining 72 h that the TER was monitored. The RTgill-W1 cell line is thus a convenient model for growing N. pemaquidensis and for studying host-pathogen interactions in AGD.     

The secretion of amylase, phospholipase and protease from Aeromonas salmonicida, and the correlation with membrane-associated ribosomes

Abstract. The following extracellular enzymes have been readily detected in the culture filtrates from Aeromonas salmonicida: amylase, phospholipase, lysophospholipase and ribonuclease. Amylase and phospholipase have been partially characterized. Evidence suggests that glycogen may be the natural substrate for amylase, and that the role of the enzyme in natural infection is to digest glycogen present in fish muscle. The secretion of amylase activity is suppressed by the addition of glucose to the growth medium. The amounts of amylase, phospholipase and protease that can be detected in culture filtrates decreases with increase in the growth temperature from 25 to 32°C. This marked decrease in secretion of hydrolytic enzymes occurs although the initial growth rates at 25 and 32°C are similar. Free and membrane associated ribosomes have been isolated from cultures grown at 25 and 32°C. At 32°C there is a smaller proportion of membrane-associated ribosomes and this is consistent with the hypothesis that extracellular enzymes from Aeromonas salmonicida are secreted on membrane-bound polysomes.     

牙鲆tyrp1a和tyrp1b的鉴定及tyrp1a与mmu-miR-143-5p_R+2的调控关系

为了研究牙鲆白化发生过程中的分子调控机制,本研究在获得正常和白化牙鲆转录组及micro RNA(mi RNA)深度测序数据的基础上,对tyrosinase related protein 1(tyrp1)和mmu-mi R-143-5p_R+2(mmu-143)进行了表达模式、靶基因预测及验证分析。首先通过RACE方法克隆得到白化相关基因tyrp1的2个转录本,进化树分析表明这2个转录本分别是tyrp1a和tyrp1b,利用RNAhybrid软件预测到mmu-143可能与tyrp1a基因存在互作关系,通过双荧光素酶实验初步验证了这一靶向关系。进一步的荧光定量结果显示,tyrp1a基因在正常牙鲆皮肤中的表达量显著高于白化牙鲆皮肤的表达量,正常牙鲆皮肤中mmu-143的表达量显著低于白化牙鲆皮肤中的表达量。本研究发现,牙鲆tyrp1存在2个转录本,分别是tyrp1a和tyrp1b。双荧光素酶实验和定量PCR分析初步证实,tyrp1a是mmu-143的靶基因,mmu-143是通过调控tyrp1a基因的表达来影响牙鲆白化的。此研究结果为深入揭示牙鲆白化发生的分子机制提供重要的基础资料。     

Median Lethal Concentration (LC50) of Piscicidal Plants and Their Utilization in Aquaculture

Aqueous and ethanol extracts of Acacia pennata (bark), Catunaregam uliginosa (fruit), Diospyros lenceofolia (fruit), and Sapindus mukorossi (fruit) were evaluated as piscicides on the freshwater fish Danio dangila, Danio rerio, and Puntius shalynius, and the air-breathing Heteropneustes fossilis. Catunaregam uliginosa (fruit) possessed relatively higher piscicidal potency followed by Sapindus mukorossi (fruit), Acacia pennata (bark), and lastly Diospyros lanceofolia (fruit). Catunaregam uliginosa can thus be used for eradicating unwanted wild fishes in a short time span at low concentrations so that the health of aquatic environment is maintained.     

The pelagic habitat analysis module for ecosystem‐based fisheries science and management

We have developed a set of tools that operate within an aquatic geographic information system to improve the accessibility, and usability of remote‐sensed satellite and computer‐modeled oceanographic data for marine science and ecosystem‐based management. The tools form the Pelagic Habitat Analysis Module (PHAM), which can be applied as a modeling platform, an investigative aid in scientific research, or utilized as a decision support system for marine ecological management. Applications include fisheries, marine biology, physical and biological oceanography, and marine spatial management. The GIS provides a home for diverse data types and automated tools for downloading remote sensed and global circulation model data. Within the GIS environment, PHAM provides a framework for seamless interactive four‐dimensional visualization, for matching between disparate data types, for flexible statistic or mechanistic model development, and for dynamic application of user developed models for habitat, density, and probability predictions. Here we describe PHAM in the context of ecosystem‐based fisheries management, and present results from case study projects which guided development. In the first, an analysis of the purse seine fishery for tropical tuna in the eastern Pacific Ocean revealed oceanographic drivers of the catch distribution and the influence of climate‐driven circulation patterns on the location of fishing grounds. To support management of the Common Thresher Shark (Alopias vulpinus) in the California Current Ecosystem, a simple empirical habitat utilization model was developed and used to dynamically predict the seasonal range expansion of common thresher shark based on oceanographic conditions.     

东莞生态园湿地鱼类群落结构

为了解东莞生态园湿地水域群落结构特征,探讨2种类型刺网(沉网和浮网)及不同网目网片渔获物组成的差异性,实验利用多网目(1.0、1.25、1.6、2.0、2.5、3.1、3.9、4.8、5.8、7.0、8.6和11.0 cm)单层刺网对该湿地的鱼类进行了调查。研究期间共采集到13种鱼类,相对重要性指数显示,海南似和莫桑比克罗非鱼为优势种,前者密度最高(占84.8%),后者生物量居多(40.2%)。渔获物物种组成沉浮网间差异显著,但单位努力捕捞数量(NPUE)、单位努力捕获重量(BPUE)及物种数与沉浮网间均无显著差异;海南似的NPUE在2种类型刺网间无显著差异,但莫桑比克罗非鱼和尼罗罗非鱼的NPUE浮网显著低于沉网。12个网目网片的渔获物物种聚成2类,1.0～2.0 cm网目网片渔获物聚为1类,海南似为指示种;2.5～11.0 cm片的聚为另1类,莫桑比克罗非鱼和尼罗罗非鱼为指示种。渔获物大小结构聚为2类,1.0～3.9 cm网目网片渔获物聚为1类,4.8～11.0 cm网目网片的聚为另1类。随着网目大小增加,渔获物平均体长线性递增,NPUE呈幂函数式下降。研究表明,为对内陆水域鱼类群落...     

Protein and energy nutrition of brook trout (Salvelinus fontinalis) at optimal and elevated temperatures

The digestible protein (DP) and digestible energy (DE) requirements for maintenance and growth of brook trout (Salvelinus fontinalis) were determined using a factorial model at either optimum (15 °C) or elevated temperature (19 °C). Several key parameters of the factorial model were measured using a series of inter‐related studies. The maintenance requirements for DP and DE were 0.10 gDP kg^?0.69 day^?1 (15 °C) and 0.31 gDP kg^?0.78 day^?1 (19 °C), and 34.86 kJDE kg^?0.84 day^?1 (15 °C) and 46.14 kJDE kg^?0.86 day^?1 (19 °C). The total requirements for DP were 0.10 gDP kg^?0.69 day^?1 + 2.14PG (protein gain) (15 °C) and 0.31 gDP kg^?0.78 day^?1 + 1.98PG (19 °C). The total requirements for DE were 36.86 kJDE kg^?0.84 day^?1 + 1.58EG (energy gain) (15 °C) and 46.14 kJDE kg^?0.86 day^?1 + 1.64EG (19 °C). The partial efficiencies for growth were 0.47 (15 °C) and 0.51 (19 °C) for protein, and 0.63 (15 °C) and 0.61 (19 °C) for energy. Nutrient gain was lower at the elevated temperature; however, feed formulation for brook trout should be adjusted to match changes in nutrient requirements at different culture temperatures. The protein and energy requirements model will be useful for developing commercial feeds and feeding charts for brook trout.     

Effects of various levels of dietary protein on survival,molting frequency and growth of juvenile blue crabs (Callinectes sapidus)

An 18-week study was conducted to examine growth characteristics of the juvenile blue crab (Callinectes sapidus Rathbun) fed various levels of dietary protein. Hatchery raised individuals (157) of identical age (160 days) from the same gravid female were assigned to one of four dietary treatments: adult brine shrimp (Artemia salina Leach) and artificial pelleted diets containing crude protein levels of 23, 37, and 49%.Growth rate, measured as a function of mean weight and mean carapace width, indicated that brine shrimp-fed crabs were larger (P < 0.05) than crabs fed artificial pelleted diets. Also, blue crabs fed either 37 or 49% crude protein were larger than crabs fed the 23% crude protein diet. Molting frequency was greater in the brine shrimp-fed crabs than in those fed the artificial diets. No differences in growth were detected when comparing males vs. females fed the artificial diets. However, females demonstrated greater growth than males within the brine shrimp treatment. Protein concentration of freeze-dried whole blue crab carcasses was greater in brine shrimp-fed crabs than in crabs fed the artificial pelleted diets (P < 0.05).     

Recruitment of sympatric vendace (Coregonus albula) and whitefish (C. lavaretus) is affected by different environmental factors

Vendace and whitefish in Lake Osensjøen (boreal south‐east Norway) were studied by means of gillnet test fishing (1976–2013) and hydro acoustic acquisition (1986–2011). Vendace increased in number between 1998 and 2009 while growth and size at maturity decreased. The relative density of whitefish decreased in the pelagic habitat, whereas growth and size remained the same. Both species exhibited varying year‐class strength. Whereas strong year‐classes of both species became less frequent after 1980 than before this, this seemed to change after 2000, especially for vendace. Generalised additive models suggested a strong positive relationship between July/August mean air temperature and year‐class strength of both vendace and whitefish. Whitefish recruitment was also negatively affected by the new regulation regime implemented since 1981, and positively correlated with water level after hatching and by late ice off. The reason for the diverging impacts of environmental factors on the two species may be that vendace spawn in deeper waters that whitefish do. The results indicate that increasing summer temperatures benefit recruitment of both species, whereas low water level and early ice off will harm whitefish recruitment. Both trends are caused by climate warming. It may be speculated that increased density of the specialised plankton feeding vendace may affect the algal community through increased predation on herbivorous zooplankton and potentially affect the trophic state of the lake.