补充CO2对光生物反应器培养新月菱形藻的影响

为了优化光生物反应器培养微藻的条件,研究了在充空气的基础补充CO2对光生物反应器培养新月菱形藻(Nitzschiaceae closterium)生长和光合作用的影响.实验表明,补充CO2(含1 000μL/L CO2的空气)促进新月菱形藻的生长,藻细胞密度和生物量显著高于对照组(CO2含量350μL/L)(P＜0.05).补充CO2也能够提高藻细胞叶绿素a和类胡萝卜素的含量(P＜0.05),但是对叶绿素b没有显著影响(P＞0.05).补充CO2能够显著提高指数生长期的最大光合速率(Pm)、光合作用效率(α)和光合作用饱和光强(Ik)(P＜0.05).结果表明,CO2是光生物反应器培养微藻的限制因子之一,补充CO2能够提高微藻的生物量.     

补充CO2对光生物反应器培养新月菱形藻的影响

为了优化光生物反应器培养微藻的条件, 研究了在充空气的基础补充CO2 对光生物反应器培养新月菱形藻(N itz schiaceae clos terium )生长和光合作用的影响。实验表明, 补充CO2 (含1 000LL /L CO2 的空气)促进新月菱形藻的生长, 藻细胞密度和生物量显著高于对照组( CO2含量350LL /L) (P < 0. 05)。补充CO2 也能够提高藻细胞叶绿素a和类胡萝卜素的含量(P < 0.05),但是对叶绿素b没有显著影响(P > 0. 05) 。补充CO2 能够显著提高指数生长期的最大光合速率( Pm )、光合作用效率( A) 和光合作用饱和光强( Ik ) (P < 0. 05)。结果表明, CO2是光生物反应器培养微藻的限制因子之一, 补充CO2 能够提高微藻的生物量。     

3种不同规模化培养模式对湛江等鞭金藻生产力及色素含量的影响

为进一步提高湛江等鞭金藻室外规模化培养效率,对管道式光生物反应器、聚乙烯桶和水泥池3种微藻室外规模化培养模式下湛江等鞭金藻的藻密度进行了评估比较.试验将接种藻密度控制为10.0×104 cells/mL,培养周期为16 d.结果显示:管道式光生物反应器培养模式下,生产力相对最高为0.0543 g/L/d、比生长速率为0...     

光照对细胞融合藻Tetraselmis sp·－1混合培养的影响

对细胞融合技术获得的1株新型融合微藻(Tetraselmis sp.-1)进行摇瓶混合培养,并分析光照强度对该培养条件下融合微藻的影响.结果表明1) 在培养基中添加葡萄糖对融合微藻的生长有强烈的促进作用,融合微藻生长速率为0. 576 g/(L*d),是自养培养的7.38倍;2)在光照强度为3 000 lx混合培养时,葡萄糖消耗速率达最大值3 g/(L*d);3)在光照强度为2 000 lx混合培养时,藻细胞密度最大,为2.60 g/L,以葡萄糖计算的基质得率随光照强度的增加而显著下降;4)融合微藻叶绿素a含量在混合培养条件下,随光照强度的增大而增加;5) 在混合培养条件下,光照对NH4+的同化为正影响,融合微藻氮含量和CC/CN分别为5.68和6.06,均介于自养培养与异养培养之间.     

Hg2+胁迫对三种微藻生长和叶绿素荧光特性的影响

研究了不同浓度的Hg2+胁迫不同时间(1～7 d)后,蛋白核小球藻(Chlorella pyrenoidosa)、斜生栅藻(Scenedesmus obliquus)和莱茵衣藻(Chlamydomonas reinhardtii)生长及叶绿素荧光特性的变化情况。测定的主要参数有光系统II(PSII)的最大光能转化效率(ΦM)、相对光合电子传递效率(rETR)、饱和光强(Ik)和细胞密度。试验结果表明,Hg2+胁迫下3种微藻的细胞密度和叶绿素荧光参数与Hg2+浓度、胁迫时间有一定的相关关系,细胞密度和叶绿素荧光各参数间有极显著的相关关系(P<0.01)。通过计算Hg2+对3种微藻不同胁迫时间的半抑制浓度(EC50),发现3种微藻对Hg2+的耐受性大小顺序为:斜生栅藻>莱茵衣藻>蛋白核小球藻。     

微藻高密度培养中的生长指标和适应机制

以雨生红球藻为材料，利用5cm光径平板式光生物反应器，在室内人工光照、通气条件下，建立高密度培养体系，探讨了微藻高密度培养中的生长指标问题。结果发现，表述微藻生长的传统指标出现失真现象。如在高密度培养生物量增加时，叶绿素含量或细胞数目指标都在一定程度上与干重指标反映的生长信息相背离，甚至出现相反的变化趋势。这主要是由于随着密度增高，细胞间的互相遮蔽作用加剧，单个藻细胞所能获得的光能逐渐减少。细胞在适应光能减弱的过程中，无论藻细胞体积大小还是单个藻细胞叶绿素含量都发生了明显变化。因此，建议评估高密度体系中微藻细胞的生长状况时，应当充分地考虑高密度效应对藻细胞生物学特性的影响，从对多种生长指标的综合分析中获得正确、客观的生长信息。同时，作者还发现微藻细胞群体和个体之间存在两种完全不同的高密度适应机制，协同适应机制和争夺机制。     

含油微藻的选育及其冷冻保藏研究

在东北地区水域采集含有浮游微藻的水样,用纱布过滤去掉水体中较大的沉淀物,取100 mL滤液转入500 mL的三角摇瓶中,加入100 L BG11培养基,在25～28℃、光照强度为3 000 Lux,光暗比为14∶10下富集培养,采用稀释涂布法分离纯化至明显的绿色。对筛出微藻在200 mL柱式光照反应器中进行光照通气培养与评价,其中生长最快的微藻生物量达到1.8305 g·L~(-1),最高的油脂含量达到34.69%,最高油脂产率达29 mg·L~(-1)·d~(-1)。取5株(一株小球藻属Chlorella、两株单针藻属Monoraphidium两株栅藻属Scenedesmus)生长快速、含油量高的藻种新鲜藻液分别离心(4 000 r·min~(-1),3 min),去离子水洗两次后,加入新鲜培养基重悬,取出部分微藻加入5种保护剂:甘油(GLY)、二甲基亚砜(DMSO)、蔗糖(SUC)、聚乙二醇(PEG)和羟二基淀粉(HES),分别置于-18℃和-80℃保藏。一个月后,取出保藏微藻,解冻,摇匀,离心,用生理盐水洗两次,去除保护剂后一部分检测细胞活性评价,一部分接种到新鲜培养基进行生长评价。选取确定保护剂后进一步考察保护剂浓度(10%、20%、30%、40%和50%)和保藏时间(3个月、6个月和9个月以及1年)。结果表明:采用二醋酸荧光素法能快速反应细胞冷冻后细胞活力,采用-18℃添加20%甘油或-80℃添加10%甘油冷冻保藏,适用于大多数含油微藻。该方法可保藏含油微藻至少1年且细胞生物量和油脂含量稳定。     

微绿球藻、隐藻、颤藻的种间竞争关系

采用陈海水配制的无机培养液,添加营养盐的无机培养液和对虾养殖池水3种培养液,分别对微绿球藻(Nannochloropsis oculata)、隐藻(Dyptomonas eyosa)和颤藻(Oscillatoria sp.)进行单培养和混合培养,探讨3种微藻的增殖规律和相互关系.观测各处理组微藻的生长状况,并以Lotka-Volterra的双种竞争模型为基础,计算3种微藻在生长拐点后各取样点的竞争抑制参数.结果显示,在各处理组中,实验前期微绿球藻和隐藻对颤藻的生长都具有一定的促进作用,颤藻在混合培养组中的生物量大于单培养组,而微绿球藻和隐藻的最大生物量均出现在其单培养组.在各组生长拐点后,微绿球藻对颤藻的影响较小,隐藻对颤藻的抑制作用明显,各取样点隐藻对颤藻的竞争参数远大于微绿球藻对颤藻的竞争参数(P＜0.05);颤藻对微绿球藻有较小的抑制作用,而对隐藻的抑制作用明显,各取样点颤藻对隐藻的竞争参数均为各组的最大值.实验结果表明,3种微藻按竞争力从大到小依次为颤藻,隐藻,微绿球藻.颤藻对隐藻和微绿球藻有较强的抑制作用,而微绿球藻和隐藻之间的竞争抑制作用较弱,能够达到"共存"状态.     

黄渤海夏季微藻调查

根据2011年夏季在黄渤海的采样调查分析了该海域网采浮游微藻的多样性,并从71个站位的采水样品与12个站位的拖网样品中分离了大量可培养的藻种。调查区拖网样品中共发现浮游微藻4门30属44种藻,以硅藻门(Bacillariophytas)为主,甲藻门(Dinophytas)次之。在实验室内利用毛细管法、平板法和稀释法分离纯化获得92株可培养微藻,经分子学鉴定为19种,包括9种硅藻、3种褐藻、3种不等鞭毛藻、2种绿藻、1种甲藻、1种定鞭藻。这些可培养微藻个体较小,多为微微型藻类和微型藻类,其中伪菱形藻(Pseudonitzschia sp.)和舟形藻(Navicula sp.)既能在固定样品中观察到,又能在实验室培养。圆筛藻(Coscinodiscus sp.)、梭角藻(Ceratium fusus)和夜光藻(Noctiluca scintillans)等小型藻类虽然在固定样品中所占比例较大,但是难以培养。此外,本次调查还首次在中国海域发现了Pseudobodo tremulans。黄渤海藻株的鉴定与培养不仅补充了中国微藻种质资源,还为促进微藻的研究和开发利用提供了重要材料。     

2种培养方式对小新月菱形藻生长及菌群结构的影响研究

文章以小新月菱形藻(Nitzschia closterium f. minutissima)为研究对象,分析比较了小新月菱形藻在负压光生物反应器与开放式桶培养下,藻密度、pH、溶解氧及菌落结构的变化情况。结果表明,在负压光生物反应器培养下的藻密度可达到1.33×10~7个·mL~(–1),明显高于开放式培养的藻密度(8.36×10~6个·mL~(–1))。藻液中pH随藻密度增加而升高,两者呈显著正相关(P0.01),在负压光生物反应器及开放式培养环境中pH最高值分别为10.3和9.3。溶解氧与pH变化趋势相反,在负压光生物反应器内溶解氧随藻密度增加而降低,最后稳定在6.5 mg·L~(–1),溶解氧的下降可能与玫瑰杆菌(Roseobacter)成为优势细菌有关。利用16S rDNA基因的高通量测序技术,分析在培养过程中藻际菌群的结构变化,发现菌落的多样性显著下降(P0.05),培养前期主要以变形杆菌(Proteobacteria)和拟杆菌(Bacteroidetes)为优势细菌,在负压光生物反应器内培养后期主要以蓝细菌(Cyanobacteria)与玫瑰杆菌为优势细菌,其菌落结构与开放式桶存在明显差异。     

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.     

美人鱼发光杆菌对凡纳滨对虾非特异性 免疫功能及抗病力的影响

在基础饲料中分别添加0.1%和1%美人鱼发光杆菌灭活菌、0.1%美人鱼发光杆菌活菌配制成3种免疫实验饲料,以基础饲料为空白对照组饲料,每组设3个平行样。对个体质量为(4.83±0.36)g的凡纳滨对虾进行为期20 d的饲养实验,分别在0、5、10、15和20d进行取样,以血清中的酚氧化酶(PO)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)、超氧化物歧化酶(SOD)和溶菌酶(UL)活性为免疫指标,探讨了美人鱼发光杆菌作为免疫制剂对凡纳滨对虾非特异性免疫效应的影响;在投喂免疫饲料后的第22天,按0.004 2 kg/kg体重的剂量,直接投喂对虾白斑综合征病毒(WSSV)病料,并记录累积死亡率。结果表明,美人鱼发光杆菌免疫实验组对凡纳滨对虾血清中PO、ACP、AKP、UL和SOD活性影响明显高于对照组,并且在饲料中添加美人鱼发光杆菌后,明显提高了对虾抵御WSSV感染的能力。其中0.1%美人鱼发光杆菌活菌实验组的抗病毒感染能力最强,WSSV感染14d内累计死亡率为63.3%±5.8%;而对照组为96.7%±3.3%。研究表明,美人鱼发光杆菌添加在对虾饲料中能提高凡纳滨对虾非特异性免疫水平,增强抵抗疾病的能力,将其作为对虾免疫增强剂具有良好的应用前景。     

Effects of Spirulina platensis as a feed additive on growth and coloration of blue dolphin cichlids (Cyrtocara moorii Boulunger, 1902)

This study was carried out to investigate the effects of replacing fish meal with dietary Spirulina as a feed supplement on the growth performance and coloration of blue dolphin cichlids (Cyrtocara moorii). Five isonitrogenous (47% crude protein) and isocaloric (17.36 kJ/g digestible energy) diets were for formulated to replace FM with 0, 5, 10, 15 and 20% Spirulina (designated as Control, SP5, SP10, SP15 and SP20 respectively) and fed to the fish (initial body weight, 3.15 ± 0.01 g). Fish were randomly distributed into fifteen 120 L aquariums (26.5 ± 1.00°C), 15 fish per aquarium. The diets were tested in triplicate for 12 weeks. Experimental groups were fed twice daily (09:00 and 17:00) by hand to satiation. At the end of the feeding trial, significantly (p < 0.05) higher weight gain (WG), specific growth rate (SGR), protein efficiency ratio (PER) and lower feed conversion ratio (FCR) were observed in fish fed the SP10 diet when compared to the SP20 diet. There was no significant difference in these parameters between the other groups. The skin coloration of blue dolphin cichlid fed a diet containing Spirulina meal was enhanced. The best coloration was observed in the SP15 group. These impressions were objectively validated by chemical determinations of carotenoids extracted from fish skins and passed statistical tests of significance. The study findings show that Spirulina meal does not diminish growth rates except at very high levels.     

Dietary thiamin and pyridoxine requirements of fingerling Indian major carp,Cirrhinus mrigala (Hamilton)

Two experiments were conducted to quantify the dietary thiamin (experiment I) and pyridoxine (experiment II) requirements of fingerling Cirrhinus mrigala for 16 weeks. In experiment I, dietary thiamin requirement was determined by feeding seven casein–gelatin‐based diets (400 g kg^?1 CP; 18.69 kJ g^?1 GE) with graded levels of thiamin (0, 0.5, 1, 2, 4, 8 and 16 mg kg^?1 diet) to triplicate groups of fish (6.15 ± 0.37 cm; 1.89 ± 0.12 g). Fish fed diet with 2 mg kg^?1 thiamin had highest specific growth rate (SGR), protein retention (PR), RNA/DNA ratio, haemoglobin (Hb), haematocrit (Hct), RBCs and best feed conversion ratio (FCR). However, highest liver thiamin concentration was recorded in fish fed 4 mg thiamin kg^?1 diet. Broken‐line analysis of SGR, PR and liver thiamin concentrations exhibited the thiamin requirement in the range of 1.79–3.34 mg kg^?1 diet (0.096–0.179 μg thiamin kJ^?1 gross energy). In experiment II, six casein–gelatin‐based diets (400 g kg^?1 CP; 18.69 kJ g^?1 GE) containing graded levels of pyridoxine (0, 2, 4, 6, 8 and 10 mg kg^?1 diet) were fed to triplicate groups of fish (6.35 ± 0.37 cm; 1.97 ± 0.12 g). Fish fed diet containing 6 mg kg^?1 pyridoxine showed best SGR, FCR, PR, RNA/DNA ratio, Hb, Hct and RBCs, whereas maximum liver pyridoxine concentration was recorded in fish fed 8 mg kg^?1 dietary pyridoxine. Broken‐line analysis of SGR, PR and liver pyridoxine concentrations reflected the pyridoxine requirement from 5.63 to 8.61 mg kg^?1 diet. Data generated during this study would be useful in formulating thiamin‐ and pyridoxine‐balanced feeds for the intensive culture of this fish.     

Sea bream culture in Egypt; status, constraints and potential

Marine fish farming in Egypt began in 1976 with the culture of gilthead sea bream, (Sparus aurata) as this fish was notably adaptable to brackish and marine pond conditions. Today, marine fish and shrimp farms amount to about 19,000 ha, out of which 42% is already in production while the rest, i.e., 58% is still under construction. In 1997, cultured gilthead sea bream production of 2,250 tons made up 3% of the 75,000 tons total aquaculture catch. In polyculture, usually with the grey mullet and sea bass, gilthead sea bream contributed 440 kg ha^–1 to the total yield of 1,700 kg ha^–1 (26%) over a period of 16 months. For the same period, the yield of monoculture ponds averaged 100 kg ha^–1, while in marine cages, yields ranged from 4–10 kg m³. In 1996–1997, fry of 0.25–1 g and fingerlings 1–10 g with a total of 3 million, were collected from the wild and 1 million fry were produced in the three marine hatcheries out of the four existing ones. The development of sea bream culture in Egypt is now severely inhibited by the shortage of seeds and adequate feeds. Exports of both sea bream and sea bass, during 1994–1996 averaged 1,300 tons per year.     

A Minchinia mercenariae‐like parasite infects cockles Cerastoderma edule in Galicia (NW Spain)

The cockle Cerastoderma edule fishery has traditionally been the most important shellfish species in terms of biomass in Galicia (NW Spain). In the course of a survey of the histopathological conditions affecting this species in the Ria of Arousa, a haplosporidan parasite that had not been observed in Galicia was detected in one of the most productive cockle beds of Galicia. Uni‐ and binucleate cells and multinucleate plasmodia were observed in the connective tissue mainly in the digestive area, gills and gonad. The parasite showed low prevalence, and it was not associated with abnormal cockle mortality. Molecular identification showed that this parasite was closely related to the haplosporidan Minchinia mercenariae that had been reported infecting hard clams Mercenaria mercenaria from the Atlantic coast of the United States. The molecular characterization of its SSU rDNA region allowed obtaining a fragment of 1,796 bp showing 98% homology with M. mercenariae parasite. Phylogenetic analysis supported this identification as this parasite was clustered in the same clade as M. mercenariae from the United States and other M. mercenariae‐like sequences from the UK, with bootstrap value of 99%. The occurrence of M. mercenariae‐like parasites infecting molluscs outside the United States is confirmed.     

A novel approach to denitrification processes in a zero-discharge recirculating system for small-scale urban aquaculture

This paper presents an innovative process to solve the nitrate build-up problem in recirculating aquaculture systems (RAS). The novel aspects of the process lie in a denitrification bioreactor system that uses solid cotton wool as the primary carbon source and a unique degassing chamber. In the latter, the water is physically stripped of dissolved gaseous O₂ (by means of a Venturi vacuum tube), and the subsequent denitrification becomes more efficient due to elimination of the problems of oxygen inhibition of denitrification and aerobic consumption of cotton wool. The cotton wool medium also serves as a physical barrier that traps organic particles, which, in turn, act as an additional carbon source for denitrification. Operation in the proposed system gives an extremely low C/N ratio of 0.82 g of cotton wool/g of nitrate N, which contributes to a significant reduction of biofilter volume. The additional advantage of using solid cotton wool as the carbon source is that it does not release organic residuals into the liquid to be recycled. Operation of the system over a long period consistently produced effluents with low nitrate levels (below 10 mg N/l), and there was only a very small need to replace system water. The overall treatment scheme, also incorporating an aerobic nitrification biofilter and a granular filtration device, produced water of excellent quality, i.e., with near-zero levels of nitrite and ammonia, a sufficiently high pH for aquaculture, and low turbidity. The proposed system thus provides a solution for sustainable small-scale, urban aquaculture operation with a very high recovery of water (over 99%) and minimal waste disposal.     

Fertilization by refrigerator stored sperm of the Indian major carp, Labeo calbasu (Hamilton, 1822)

This study reports on the spermatological properties, and on the development of a protocol for refrigerator storage (4°C) of Labeo calbasu (Hamilton, 1822) sperm for artificial breeding. Volume, motility, concentration and pH of the freshly collected sperm were 2.21 ± 0.53 (μL g^?1 of fish weight) (mean ± SD), 95 ± 1 (%), 1.93 ± 0.44 × 10⁹ (cells mL^?1) and 7.56 ± 0.17 respectively. Sperm activation was evaluated at different osmolalities of NaCl solution, and motility ceased completely when osmolality of the extender was ≥287 mOsmol kg^?1. Sperm retained motility for 24, 72 and 108 h, after refrigerator storage when sperm were undiluted, suspended in Alsever's solution and suspended in Alsever's solution containing 5% methanol respectively. Fertilization rate of fresh eggs with sperm stored at 4°C in Alsever's solution and Alsever's solution containing 5% methanol was 77% and 60% with a hatching rate of 60% and 43% respectively. The fertilization and hatching success of the stored sperm suggests potential to use refrigeration for transporting genetic material to hatcheries for artificial breeding of L. calbasu in Bangladesh.     

Effects of metomidate anaesthesia or transfer to pur sea water on plasma parameters in ammonia-exposed Atlantic salmon (Salmo salar L) in sea water

Atlantic salmon (Salmo salar L) postsmolts weighing 150 ± 53 g were exposed to 14–15 mg l^–1 TA-N (total ammonia-N) in sea water in 1 m³ tanks for 24h. Blood samples were then taken A) immediately after the fish were netted from the exposure tanks and stunned by a blow to the head; B) 2–20 min after the fish were transferred to 15 l of an anaesthetic solution of metomidate in ammonia-free sea water; or C) 2–20 min after the fish were transferred to 15 l of ammonia-free sea water. Plasma TA-N level was 18% lower in the anaesthetised fish compared to in the fish sampled directly from the exposure tanks (p 0.05), and accordingly 16% lower in the fish transferred to pure sea water although this difference was not significant (p = 0.07). Plasma glucose level was higher in the fish transferred to pure sea water than in the fish receiving the two other treatments (p 0.05), but plasma urea, osmolality, Na^{+, Cl–, Ca2+ or Mg2+} levels did not vary significantly between the different treatments. Plasma TA-N level increased with time in the fish in the metomidate solution (p 0.02).