Biocontrol of Vibrio vulnificus strains challenged with Isochrysis galbana cultures

Journal of Applied Phycology - The pathogenic bacterium Vibrio vulnificus is related to human infections by direct contact with the bacteria or by consuming raw aquacultural products, like oysters...     

A study of the assimilation of fluorescent pigments of microalgae Isochrysis galbana by the early larval stages of turbot and herring

The appearance of large supranuclear vacuoles in the enterocytes of 1- and 4-days post-hatch larvae of turbot and herring, respectively, revealed by the pinocytotic uptake of a fluorescent marker (FITCdextran), indicates a potential for the absorption of dissolved nutrients by the endotrophic stages of marine fish larvae. Ingestion of algal cells by turbot larvae was observed soon after hatching, but low level pinocytotic absorption of algal material was first seen during the second day. More extensive lysis of algal cells and pinocytotic absorption occurred 24 h later. Although lysis of I. galbana was shown to occur at low external osmolarities, it is unlikely that sufficiently low osmolarities present in the hind gut of turbot larvae explained the observed rupture of algae. Other mechanisms for digestion are discussed. In newly hatched herring larvae, algal cells were unable to pass beyond the constriction in the mid-gut caused by the yolk sac. When algal cells were eventually seen to pass into the hind gut, there was no evidence of algal digestion or absorption throughout the remaining endotrophic and early exotrophic stages of herring larvae, although pinocytosis was observed to occur mid-way through the endotrophic stage.     

Growth inhibition of the eight species of microalgae by growth inhibitor from the culture of Isochrysis galbana and its isolation and identification

A type of growth inhibitor was successfully isolated and purified from cell-free filtrates of cultural medium at the death phase of Isochrysis galbana, and its chemical structure was confirmed by the methods of FABMS, UV, ¹H-NMR, ¹³C-NMR and 2D NMR, which was 1-[hydroxyl-diethyl malonate]-isopropyl dodecenoic acid, C₂₂H₃₈O₇. The results showed that the growth-inhibitor strongly inhibited the growth of Isochrysis galbana, and the growth of the eight species of microalgae (Dunaliella salina, Platymonas elliptica, Chlorella vugralis, Nitzschia closterium, Chaetoceros muelleri, Chaetoceros gracilis, Nitzschia closterium minutissima, Phaeodactylum tricornutum) also could be regulated by the growth-inhibitor in a concentration-dependent manner. The further investigation found that the synthesis process of chlorophyll and protein in the cells of all test microalgae could be inhibited by the growth inhibitor, and the content of chlorophyll and protein significantly decreased.     

Docosahexaenoic acid ethyl esters from Isochrysis galbana

In order to produce docosahexaenoic acid (DHA), a culture of the microalgal strain Isochrysis galbana was implemented. In Erlenmeyer flasks, a natural seawater medium, the Provasoli 1/3 medium, was compared to the classical Jones medium for DHA production. The Provasoli 1/3 medium stimulated growth (0.44 d(-1)), but influenced DHA accumulation negatively (0.240 pg cell(-1)). However, DHA production per liter of culture medium were of the same order of magnitude with both media (0.961 mg l(-1)). In a 2-l bioreactor, DHA production per liter of culture medium did not increase significantly between 4 and 8 days of culture. With a view to optimize DHA productivity, cells should be harvested at the end of exponential phase i.e. after 4 days of culture. Two strategies were then attempted to produce DHA ethyl esters. First, lipids from I. galbana were submitted to lipase-catalyzed transesterification with ethanol. Secondly, fatty acids from I. galbana were submitted to lipase catalyzed esterification with ethanol. In both cases, lipase from Candida antarctica was shown to be the best candidate, among the five tested, with conversion yields of 20 and 60% after 24 h of transesterification and esterification respectively.     

Feeding kinetics of Brachionus plicatilis fed Isochrysis galbana

Clearance and ingestion rates of Brachionus plicatilis were measured using ¹⁴C-labeled Isochrysis galbana Tahiti. Experiments were conducted at 20–22 °C, 20 ppt salinity, and algal concentrations ranging from 0.13–64 mg C 1^–1. Clearance rates were constant and maximal at concentrations <2 mg C 1^–1, with maximum rates ranging from 3.4–6.9 µl ind.^–1 hr^–1. The ingestion rate varied with food concentration, and was described by a rectilinear model. The maximum ingestion rate varied considerably, and was dependent on the growth rate of the rotifers. Depending on the pre-conditions, B. plicatilis ingested about 0.5 to 2 times its body carbon per day at saturating food concentrations.     

Larval growth and survival of Echinometra lucunter (Echinoidea: Echinometridae) fed with microalgae Chaetoceros gracilis and Isochrysis galbana

Thirty sexually mature sea urchins (Echinometra lucunter; diameter 45.8 +/- 17.5 mm) were collected at Macanao, Margarita Island, Venezuela (11 degrees 48'29" N / 64 degrees 13'10" W). They were injected potassium chloride (50 M) directly into the celomic cavity. After two minutes 90% spawned (17 females and 10 males), the others never spawned. Fertilization was 87.0 +/- 12.6% (1:100 oocytes/sperm) at 29 +/- 2 degrees C. The fertile eggs were placed in three treatment gropsu with nine containers (18 liters; 2 eggs/ml) each, all with bottom aeration. Treatments were: Chaetoceros gracilis; Isochrysis galbana, and a mixture of both microalgae (respectively: 20 000 and 60,000 cell/ml for each microalgae, 1:1 for the mixture). Salinity, pH, temperature and larval survival were determinated daily. The study ended when the post-metamorphic phase was completed. The embryonic development time was 16.3 +/- 0.2 h until the prism stage at pH 8.4 +/- 0.1; 38 +/- 1 psu and 28 +/- 1.4 degrees C. The two-arms larval stage was reached at 24 h: 33 min, with a total length of 190 +/- 16.3 microm fed on C. gracilis, 152 +/- 19.0 microm with I. galbana and 182.4 +/- 14.1 microm with the mixture. The larvae next to metamorphosis reabsorbed the arms and had the characteristic shape of juvenile urchins at 12 days with 670.2 +/- 22.2 microm fed on C. gracilis, 665 +/- 12.1 microm fed on I. galbana and 670 +/- 14.1 microm fed on the mixture. The accumulated survival to the juvenile stage was 14.7 +/- 3.8% when fed on C. gracilis, higher than the other treatments (5.4 +/- 1.2; 14.0 +/- 2.6). E. lucunter is an excellent prospect to be commercially cultured because of its short embryonic (16 hours) and larval development time (12 days) and good survival rate when fed on monoculture (C. gracilis and I. galbana) or mixed diet (we recommend C. gracilis).     

球等鞭金藻生长抑制物对藻细胞的抑制效应及抗氧化剂对抑制效应的抵制作用

以球等鞭金藻为材料,研究生长抑制物GI对藻细胞生长、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化物酶(peroxidase,POD)活性和丙二醛(malondialdehyde,MDA)含量的影响;同时研究4种抗氧化剂(抗坏血酸、柠檬酸、乙二胺四乙酸二钠和3-叔丁基-4-羟基-苯甲醚)对GI抑制效应的抵制作用.结果表明,0.10mg/LGI处理组藻细胞密度、SOD和POD活性明显低于对照组,而MDA含量明显高于对照组.且随着GI浓度的继续增大,细胞密度、SOD和POD活性急剧降低,而MDA含量进一步升高.GI浓度为0.30mg/L时,处理组藻细胞密度、SOD和POD活性以及MDA含量分别为对照组藻细胞密度的0.05倍、SOD活性的0.56倍、POD活性的0.59倍和MDA含量的2.2倍.4种抗氧化剂均能有效地抵制GI对藻细胞的抑制效应,使细胞密度、SOD活性和POD活性提高,MDA含量降低.添加抗氧化剂处理组的藻细胞密度为对照组细胞密度的1.38～1.90倍、SOD和POD活性分别为对照组活性的1.49～2.12倍和1.55～2.13倍,而MDA含量比对照组含量降低57.7%～87.9%.生长抑制物的胁迫使球等鞭金藻细胞体内积累了过量的活性氧,而抗氧化剂通过清除藻细胞体内积累的活性氧,减轻了膜脂过氧化伤害,从而抵制了生长抑制物对藻细胞的抑制效应.     

三种絮凝剂对球等鞭金藻絮凝作用

以3种絮凝剂对等鞭金藻的采收效果及其对藻体的影响为研究目标,以分光光度法、重量法以及显微镜观测为主要研究方法,测定了絮凝剂对藻细胞的絮凝效率、藻体总脂含量的影响以及藻细胞形态变化。结果表明:氯化铁和明矾的絮凝速率最快(<4h);氯化铁浓度≥20mg.L-1,明矾浓度≥80mg.L-1时,可以絮凝沉淀90%以上藻体;比较藻体的损伤程度和总脂产率发现,明矾浓度为80mg.L-1时,藻体总脂产率的最高,达29.9%,并且对细胞伤害最小;可采用80mg.L-1明矾作为絮凝剂对球等鞭金藻进行采收,为生物柴油制备生产提供基础原料。     

Growth activation of the microalga Isochrysis galbana by the aqueous extract of the seaweed Monostroma nitidum

Cell growth of the marine microalga Isochrysis galbana was regulated by the addition of seaweed extracts in the culture medium. Methanol-soluble extracts from 27 species of seaweed showed growth activation only from Enteromorpha linza, and growth inhibition from Ishige foliacea and Sargassum sagamianum. Water-soluble extracts from Grateloupia turuturu and Monostroma nitidum showed growth activation, while none of the seaweed showed growth inhibition. From results of growth activation of extracts on I. galbana, the water extract of M. nitidum was the most effective up to two-fold increase in cell density with the addition of 1 mg mL^-1 of extract to the medium. The cell growth rate was increased from 0.52 to 0.65 d^-1. Cell size, gross biochemical compositions, fatty acid compositions, and digestion efficiency by shellfish differed marginally between cultures of I. galbana grown with and without the M. nitidum aqueous extract. This extract has also enhanced the growth of other feed microalgae tested, including Dunaliella salina. This revised version was published online in August 2006 with corrections to the Cover Date.     

Growth and pigment accumulation in nutrient-depleted Isochrysis aff. galbana T-ISO

The effect of three different nutrient depletions (nitrogen, sulphur and magnesium) on the growth and pigment accumulation of the haptophyte Isochrysis aff. galbana (clone T-ISO) has been studied. Pigments were quantified based on RP-UHPLC-PDA-MSⁿ analysis. All nutrient depletions led to reduced maximal biomass concentrations. Besides, all nutrient-depleted cultures accumulated 3-hydroxyechinenone. To our knowledge, this is the first time that 3-hydroxyechinenone has been found in I. aff. galbana T-ISO. Most 3-hydroxyechinenone, as well as the most echinenone and diatoxanthin, were found in the nitrogen-limited culture in which a more severe limitation resulted in higher cellular contents. Similar to accumulation of diatoxanthin, accumulation of 3-hydroxyechinenone and echinenone may be part of a global (stress) response mechanism to oversaturating light conditions.     

智舌快速鉴定水产品多种致病性弧菌

【目的】探索研究能很好显示被检测液态食品样品综合信息的电子舌是否也能很好的显示致病性弧菌液体培养物的综合信息,籍此探讨一类致病性弧菌的快速鉴定鉴别新技术。【方法】基于多频脉冲伏安法的智舌,结合主成分分析,对源自水产品的11种致病性弧菌进行区分鉴定研究,得到最佳电极阵列和频率段组合。【结果】结果显示,区分致病性弧菌效果好的电极和频率段分别是:钛电极的100Hz、银电极的100Hz和钨电极的1Hz、10Hz。钨电极的1Hz频率段能够独立的把11种致病性弧菌在同一张主成分得分图上区分开;钛电极的100Hz、银电极的100Hz和钨电极的10Hz两两组合也能把11种致病性弧菌主成分得分图上区分开。【结论】研究结果表明,智舌伏安法结合主成分分析法区分致病性弧菌是可行的,具有很大的持续研究价值,有望发展成一类很有发展优势的快速鉴定致病性弧菌乃至延伸至其他致病菌的新技术。     

NaNO3浓度对球等鞭金藻生长及所含脂肪酸的影响

用NaNO3作氮源,分别作了氮浓度的5个水平对球等鞭金藻H29的相对生长速率、总脂含量及脂肪酸组成的影响实验。实验结果显示,H29的相对生长率和总脂含量随N浓度的增加而增加,在897.6 mg/L时总脂含量达到最大值(23.4%)。而DHA(22:6n-3)和PUFAs(polyunsaturated fatty acids)含量随氮浓度的改变有较大变化,DHA含量总体随N浓度增加而下降,在较低氮浓度(74.8mg/L)时含量达到最大值12.11%。     

Predator-prey interactions between Isochrysis galbana and Oxyrrhis marina. II. Release of non-protein amines and faeces during predation of Isochrysis

During the growth of Isochrysis galbana, several non-proteinamines may be detected in the growth medium. Of these, one (termed‘TTl’) accumulates in proportion to the numbersof cells present. The concentrations of ‘TTl’, andof another (termed ‘TA’), are 3–5 times higherin cultures in which Isochrysis is predated by Oxyrrhis marina.The lowest estimates of the concentration of extracellular ‘TTl’are an order of magnitude higher than those of any protein aminoacid. Of the protein amino acids, some like glycine are utilizedduring predation while others, like histidine, accumulate inthe medium Because of the unknown N-content and reactivity ofthe non-protein amines during HPLC, it is not possible to sayif these compounds (together with other components of dissolvedorganic N) form a significant proportion of the unaccountedfor N in the system after predatory activity. During predationin the absence of detectable free ammonium (when Isochrysismay be expected to be N-deprived), particles accumulate in themedium. Most of these are <2.5 µ.m in diameter andare suggested to be remains of digested prey. There is evidenceof a reassimilation of these particles by prey-deplete Oxyrrhis.     

Biomass production and biochemical variability of the marine microalga Isochrysis galbana in relation to culture medium

We have studied the autotrophic growth of the marine microalga, Isochrysis galbana Parke, in a batch photobioreactor, comparing five different culture media and analysing the influence of each on growth kinetics as well as on the fatty-acid composition and protein content of the biomass. All the experiments were performed at 15 degrees C, with the culture medium at pH 8.0, a specific rate of air supply of 1vv(-1)min(-1) and a continuous illumination of 40-43Wm(-2). The results show no parallel between good nutritional characteristics and high values of the kinetic parameters. Nevertheless, a compromise between the nutritional factors and growth kinetics could be provided by Ukeles medium, which provided a biomass with a good composition in polyunsaturated fatty acids (quotient n3/n6=3.2), an adequate protein content (25.3%) and relatively high values, although not the highest registered, for maximum specific growth rate (μ(m)=0.018h(-1)) and biomass productivity (1.9x10(-3)kgm(-3)h(-1)).