斜带石斑鱼肝细胞分离及原代培养方法的建立

本研究以斜带石斑鱼肝细胞为实验对象,在不同培养条件下进行原代培养,旨在探讨稳定可靠的斜带石斑鱼肝细胞分离及原代培养方法。采用组织块分离法和胰蛋白酶(含EDTA)消化法分离肝细胞,并通过密度梯度离心法分离纯化肝细胞,细胞悬液于DMEM/F-12、M199和L-15培养液中培养;细胞活力及数量采用血球计数板计数,并通过MTT法测定细胞增殖率;同时,测定不同时间培养上清液中乳酸脱氢酶(LDH)活性、白蛋白(ALB)和尿素氮(BUN)的含量,以分析肝细胞生长状态。结果表明,组织块方法不适于斜带石斑鱼肝细胞的培养,未见细胞从组织块中迁出,而胰蛋白酶消化法获得良好稳定的培养效果,细胞产量达到1.6×108个/g肝重,活细胞数达到95%;L-15培养基细胞生长明显优于DMEM/F-12和M199培养基;启动原代培养的48~72 h阶段肝细胞生长代谢旺盛,培养上清液中LDH活性显著降低,ALB和BUN含量显著升高。结果显示,0.25%的胰蛋白酶常温消化法适合斜带石斑鱼肝细胞的分离,斜带石斑鱼肝细胞原代培养的最适培养基为L-15培养基,肝细胞在启动原代培养的48~72 h生长代谢旺盛。     

厦门贝类养殖区沉积物中重金属的污染水平及其潜在生态危害

根据2005年4月和10月厦门贝类养殖区环境调查资料,着重对贝类养殖区表层沉积物中重金属的含量水平进行分析,并对贝类养殖区的表层沉积物进行质量评价及潜在生态危害评价.结果表明:厦门贝类养殖区沉积物中Cu、Pb、Cd、Zn、Hg、As的平均含量分别为20.0mg.kg-1,39.5mg.kg-1,0.07mg.kg-1,97.3mg.kg-1,0.05mg.kg-1,7.20mg.kg-1.其中同安湾北部和湾顶养殖区及西海域的杏林、东山养殖区,6种元素的含量均较高;位于同安湾的刘五店、澳头养殖区,含量均较低.沉积物质量评价结果表明,贝类养殖区沉积物中重金属的含量符合第一类海洋沉积物质量标准,属于较清洁水平.Hg、As和Cd的潜在生态危害系数大于Cu、Pb、Zn,整个养殖区沉积物中重金属为轻微生态危害,养殖环境良好.     

草鱼3个6-磷酸葡萄糖酶催化亚基的基因表达分析及高糖饲料对其表达的影响

为了探讨6-磷酸葡萄糖酶催化亚基(glucose-6-phosphatase catalytic subunit,G6PC)在草鱼(Ctenopharyngodon idellus)糖代谢中的作用,采用同源序列比对的方式,在草鱼基因组中获取了3个g6pc基因的序列,通过序列比对和进化树分析,将其分别命名为g6pca、g6pcb1和g6pcb2,其编码的氨基酸序列与斑马鱼(Danio rerio)、虹鳟(Oncorhynchus mykiss)和人(Homo sapiens)具有较高的同源性,相似度分别为85%～94%、64%～83%和54%～66%。同线性分析表明g6pc在草鱼染色体上的分布与斑马鱼等高度相似,表明草鱼g6pc基因在进化中具有较高的保守性。利用RT-PCR检测3个基因在鳃、脂肪、脑、心脏、肝、肾、前肠、中肠、后肠和肌肉10个组织中的表达,结果显示, g6pca在脑和肝中表达量较高,脂肪组织次之; g6pcb1在肝中表达量最高,中肠次之; g6pcb2在心脏表达量最高,其次为脂肪组织。同时探讨了高糖饲料对草鱼不同g6pc亚型转录水平的影响,以及不同浓度葡萄糖和胰岛素刺激草鱼肝细胞(L8824)后对g6pca转录水平的影响。结果显示,饲喂高糖饲料(7周)后,与对照组相比,草鱼肝脏g6pca mRNA水平显著升高, g6pcb1和g6pcb2 mRNA水平无显著变化。离体情况,与5 mmol/L葡萄糖组相比,15 mmol/L葡萄糖显著增加了L8824的g6pca mRNA水平,且1 mol/L胰岛素可以抑制这种作用; 30 mmol/L葡萄糖对L8824 g6pca mRNA水平无显著性影响。本研究表明,草鱼g6pc发生加倍后存在功能分化,高糖可以诱导g6pca mRNA的表达,而对g6pcb1和g6pcb2的转录水平无影响,其具体功能还需进一步研究。     

洪湖低湖田改造池塘河蟹养殖技术研究

在洪湖湖滨地区筛选了4种当地主要的河蟹(Eriocheir sinensis)养殖方式,对池塘与水源的环境因子进行了监测比较,分析各因子对河蟹收获参数的影响。结果显示:7—10月中,8月水温和氨氮含量最高,各池塘水体理化指标月平均值无显著差异;各池塘水草生物量逐月急剧下降,至10月底基本消失,其月平均值与成蟹体重、回捕率及特定生长率显著负相关。高温季节是防止河蟹氨氮中毒死亡的关键时期;水草调节水体营养、稳定底质、吸附水体悬浮质的功能对河蟹存活及生长有重要作用。当地优势池塘养殖方式为:放养规格约5 g/ind.,密度16875 ind./ha,水草移栽以伊乐藻为主,搭配其它种类,覆盖率约为60%,动物性饵料投喂比例约为66%,螺蛳投喂量约3638 kg/ha。     

From small-scale habitat loopholes to decadal cycles: a habitat-based hypothesis explaining fluctuation in pelagic fish populations off Peru

The Peru‐Humboldt Current system (HCS) supports the world's largest pelagic fisheries. Among the world's eastern boundary current systems, it is the most exposed to high climatic stress and is directly affected by El Niño and La Niña events. In this volatile ecosystem, fish have been led to develop adaptive strategies in space and time. In this paper, we attempt to understand the mechanisms underlying such strategies, focusing on the El Niño 1997–98 in Peru from which an extensive set of hydrographic, capture and acoustic survey data are available. An integrated analysis of the data is crucial, as each has substantial shortcomings individually; for example, both catch data and acoustic surveys may easily lead to wrong conclusions. Existing hypotheses on anchovy and sardine alternations lead us to a ‘habitat‐based’ synthetic hypothesis. Using our data, an integrated approach evaluated how fish responded to habitat variation, and determined the consequences in terms of fish‐population variability. Various factors occurring at a range of different spatio‐temporal scales were considered: interdecadal regime (warm ‘El Viejo’/cool ‘La Vieja’ decadal scale); strength and the duration of the El Niño Southern Oscillation event (interannual scale); population condition before the event (interannual scale); fishing pressure and other predation (annual scale); changes in reproductive behaviour (intra‐annual scale); presence of local upwelling (local scale). During El Niño 1997–98, anchovy was able to exploit a small‐scale temporal and spatial ‘loophole’ inside the general unfavourable conditions. Moreover, sardine did not do better than anchovy during this El Niño and was not able to take advantage of the ‘loophole’ opened by this short‐term event. Our results question the traditional view that El Niño is bad for anchovy and good for sardine.     

南海区驯养条石鲷亲鱼的初次性成熟和产卵

2008年4月初在广东省饶平县对2006年5月人工孵化和培育的2龄条石鲷（ Oplegnathus fasciatus）成熟亲鱼进行激素诱导,研究和观察条石鲷亲鱼初次性成熟的繁殖生物学。结果显示,在南海区全人工养殖的条石鲷亲鱼初次性成熟年龄为2龄,成熟亲鱼的最小型为全长245 mm、体质量610 g,最大个体为全长300 mm、体质量1450 g;产卵季节为4月10日~7月15日,产卵盛期为4月中旬至6月下旬;为升温产卵型鱼类,产卵温度为20.0~28.8℃,适宜的产卵温度为20.7~27.6℃;雌、雄亲鱼发育同步,个体大小和成熟年龄差别不大;雌鱼属于一年一次分批产卵类型,产卵期超过3个月;24尾初次性成熟雌性亲鱼的总产卵量为3180.0×104粒,日最高产卵量为341.5×104粒;其产卵量和受精卵质量与水温的变化关系密切,受精卵的浮卵率随温度的变化而变化,在水温相对衡定时,浮卵率相对稳定并维持在80%~95%的较高水平;受精卵平均卵径为（0.860±0.023）mm,油球径为（0.191±0.009）mm。     

绿原酸诱导凡纳滨对虾抗氧化功能及抵御低盐度胁迫的响应

选择健康、均匀, 体质量为(6.74 ± 0.08) g的凡纳滨对虾(Litopenaeus vannamei),  将360尾随机分为4组, 分别投喂含有0 mg/kg、100 mg/kg、200 mg/kg和400 mg/kg绿原酸的饲料28 d, 随后将对虾从盐度为32的天然海水直接转移至盐度为10的海水中暴露72 h, 测定对虾肝胰腺TAS、GPx、CAT活力和GPx、CAT基因表达水平。结果显示, 在天然海水养殖条件下, 绿原酸对凡纳滨对虾肝胰腺TAS、GPx活性无明显影响, 但投喂含有绿原酸的饲料28 d, 对虾肝胰腺GPx、CAT和GPx、CAT基因表达水平显著高于D0组(P<0.05), 以D2组GPx活性和GPx、CAT基因表达水平最高, 分别为164.29 U/mgprot和1.61、2.14倍。低盐度胁迫24 h, D0组对虾抗氧化反应表现在TAS、GPx活性和GPx基因表达水平较28 d显著升高(P<0.05), 分别增加了31.30%、27.96%和170%, 而绿原酸各组对虾肝胰腺TAS、GPx活性和GPx基因表达水平显著低于D0组(P<0.05), 说明绿原酸可有效缓解低盐度胁迫下对虾抗氧化系统酶活性的剧烈波动。低盐度胁迫72 h, 绿原酸各组对虾肝胰腺TAS、GPx、CAT活性及CAT基因表达水平均明显高于D0组。上述结果表明绿原酸能够诱导凡纳滨对虾的抗氧化系统功能, 在凡纳滨对虾抵抗低盐度胁迫中发挥着重要的作用。本研究旨为开发绿色植物提取物在对虾养殖中的应用, 解决盐度胁迫对机体造成的氧化损伤提供理论依据。

     

Soybean molasses as an organic carbon source in the farming of Litopenaeus vannamei (Boone, 1931) in a biofloc system

Soybean molasses was evaluated as a partial replacement for sugarcane molasses as a carbon source for biofloc development in the superintensive culture of Pacific white shrimp (Litopenaeus vannamei). A 50‐day study was conducted with juvenile (3.2 g) shrimp stocked in 16 800 L tanks at a stocking density of 250 shrimp m^?3. Control of total ammonia concentration was performed by the addition of combined mixtures of soybean and sugarcane molasses to the culture water. Three different molasses treatments were evaluated using different soybean‐to‐sugarcane molasses ratios: 15–85%, 38–62% and 60–40% respectively. The control group was treated only with sugarcane molasses. Water quality, chlorophyll a concentration, heterotrophic bacterial load, Vibrio spp. concentration and zootechnical indexes were all evaluated. Total ammonia concentration was controlled by heterotrophic and chemotrophic pathways. Biofloc formation, as quantified by measuring the total suspended solids, was not altered. The Vibrio spp. concentration showed a significant reduction in treatments with soybean‐to‐sugarcane molasses ratios of 38–62% and 60–40%. All combined mixtures of soybean and sugarcane molasses could maintain water quality and productivity in the superintensive culture of L. vannamei using the biofloc system. Thus, the potential use of a residue from agroindustry as a carbon source in a biofloc culture is demonstrated.     

Relationship Between Dietary Protein Source and Feeding Frequency During Feeding Nile Tilapia,Oreochromis niloticus (L.) Cultured in Concrete Tanks

ABSTRACT

A 24-week feeding trial was conducted in concrete tanks with Nile tilapia, Oreochromis niloticus (L.) with an average initial weight and length of 50.87 ± 6.03 g and 14.4 ± 0.45 cm, respectively, to examine the effect of two protein sources and two feeding frequencies on growth performance, production traits, and body composition. Twelve 4 m³ concrete tanks (2 × 2 × 1.25 m, long, width, and height) were each stocked with 100 fish and fed diets containing either fish meal protein or soybean meal protein at two feeding frequency of two times daily or four times daily (2 × 2 × 3 factorial experiment). The results revealed that there was no significant effect of dietary protein sources on growth rate, whereas there was a significant increase in growth rate with increasing feeding frequency (P < 0.05). The same trend was also observed for mean body weight (g), production rate (kg/m3), specific growth rate (SGR % day^?1). The best final mean body weight (g), specific growth rate (SGR % day^?1), and production rate (kg/m3) were recorded in groups of fish fed with feeding frequency four times daily. Whole fish moisture, protein, fat, and ash contents were significantly influenced (P ≤ 0.05) by protein sources and feeding frequency, except protein not influenced with feeding frequency. Energy was significantly (P < 0.05) influenced by feeding frequency, but not by dietary protein sources. Economic evaluation indicated that soybean meal (Diet B) at four times feeding daily was the most cost-effective and affordable feed strategy for farmers. We conclude that a soybean diet fed four times daily is recommended for adult Nile tilapia reared in concrete tanks.     

变频调速装置在给水系统中的应用分析

分析讨论了工业企业给水系统的特点和当前常用的给水系统调节方式和优缺点及变频调速装置的节能原理,指出变频调速装置应用于给水系统是最为节能的运行方式.