The effect of cell density on biomass and fatty acid productivity during cultivation of Rhodomonas salina in a tubular photobioreactor

The microalga Rhodomonas salina is widely used in aquaculture. There is a need for optimization of the growth of the microalgae and its content of essential fatty acids. Here, the fatty acid profile of Rhodomonas in relation to cell density during cultivation in a tubular PBR is investigated. It is expected that cell density is an important factor in controlling productivity and fatty acid content of the microalgae because cell density is important in determining light availability due to the self‐shading of the algae. The carbon productivity as a function of cell density is described by a saturation curve. The carbon productivity and the productivity of total fatty acids are lowest at the lowest cell density, and independent of cell density at higher cell densities. The relative contribution of the two poly‐unsaturated fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) increases with increasing cell density and saturates at 1 × 10⁶ cells/ml. We conclude that large‐scale production of Rhodomonas in this tubular PBR should take place at cell densities of 1 × 10⁶ cells/ml, while there are indications for increasing difficulties in maintaining steady‐state production in this PBR at higher cell densities.     

MALDI-TOF MS identification of Prototheca algae associated with bovine mastitis

We evaluated the use of MALDI-TOF MS for the identification of 3 major, dairy-associated Prototheca species, namely, Prototheca bovis (formerly P. zopfii genotype 2), P. blaschkeae, and P. ciferrii (formerly P. zopfii genotype 1). The MALDI-TOF MS spectra established for those species were introduced into the reference spectra library of the Bruker Biotyper MALDI-TOF MS analysis software. Next, 31 Prototheca isolates from Holstein cows with mastitis, from herds located in the midwestern area of São Paulo State, Brazil, were subjected to MALDI-TOF MS profiling. MALDI-TOF MS allowed identification of 22 of 27 P. bovis and 3 of 4 P. blaschkeae isolates with scores >2.0, with 5 of 27 P. bovis and 1 of 4 P. blaschkeae isolates identified only to the genus level. With our extended algae database, MALDI-TOF MS can contribute to quick and effective speciation of Prototheca from mastitis cases.     

Laminarin,a Major Polysaccharide in Stramenopiles

During the processes of primary and secondary endosymbiosis, different microalgae evolved to synthesis different storage polysaccharides. In stramenopiles, the main storage polysaccharides are β-1,3-glucan, or laminarin, in vacuoles. Currently, laminarin is gaining considerable attention due to its application in the food, cosmetic and pharmaceuticals industries, and also its importance in global biogeochemical cycles (especially in the ocean carbon cycle). In this review, the structures, composition, contents, and bioactivity of laminarin were summarized in different algae. It was shown that the general features of laminarin are species-dependence. Furthermore, the proposed biosynthesis and catabolism pathways of laminarin, functions of key genes, and diel regulation of laminarin were also depicted and comprehensively discussed for the first time. However, the complete pathways, functions of genes, and diel regulatory mechanisms of laminarin require more biomolecular studies. This review provides more useful information and identifies the knowledge gap regarding the future studies of laminarin and its applications.     

Digestibility of the defatted microalgae Nannochloropsis sp. and Desmodesmus sp. when fed to Atlantic salmon,Salmo salar

The aim of the study was to investigate the apparent digestibility coefficient (ADC) of defatted biomass derived from microalgae Nannochloropsis sp. and Desmodesmus sp. when fed to Atlantic salmon postsmolts in seawater. Two experiments, one employing cold‐pelleted and the other employing extruded pellets, were carried out to determine the ADC of dry matter (DM), protein, ash and energy. The test feeds consisted of a fishmeal‐based reference feed and the algal biomass, at a ratio of 70:30, and yttrium oxide as the inert marker. The ADC of DM and protein in fish fed microalgae‐incorporated cold‐pelleted feeds were significantly higher for Nannochloropsis sp. compared with Desmodesmus sp. The Nannochloropsis sp. inclusion in extruded feeds also led to higher ADCs of DM and energy compared with Desmodesmus sp. The extrusion processing significantly increased the ADC of ash in both defatted microalgae biomass, as well as ADCs of DM in Nannochloropsis and protein in Desmodesmus sp. In conclusion, the microalga Nannochloropsis sp. was more digestible than Desmodesmus sp., and extrusion processing can be used as a means to improve digestibility of certain nutrients.     

Effects of dietary docosahexaenoic acid sources,microalgae meal and oil,on growth,fatty acid composition and docosahexaenoic acid retention of orange‐spotted grouper,Epinephelus coioides

This study evaluated the effects of Aurantiochytrium spp. microalgae meal and oil as dietary docosahexaenoic acid (DHA) sources on the growth, fatty acid composition and DHA retention of orange‐spotted grouper, Epinephelus coioides. Dietary fish oil was replaced with microalgae meal or oil to provide an equal amount of DHA as a fish oil‐containing basal diet. In total, three experimental diets were fed to triplicate groups of fish (initial wt: 8.48 ± 0.06 g) in a recirculating system for 8 weeks. The weight gain and feed efficiency of the fish did not differ significantly among the experimental diets. The fatty acid composition of the whole body of the fish generally reflected the composition of their diet. The concentration of eicosapentaenoic acid in the whole body was higher in the fish fed the fish meal control diet than in those fed the two experimental diets The fish fed the control diet and those fed the diet containing microalgae oil exhibited higher DHA concentrations than did the fish fed the diet containing microalgae meal. The whole‐body DHA retention was the highest in the fish fed the diet with microalgae oil, followed by the fish fed the control diet. The lowest whole‐body DHA retention was observed in the fish fed the diet containing microalgae meal. The results suggested that the oil from Aurantiochytrium spp. microalgae can be used as DHA source for the grouper. DHA utilization by the fish was higher when the diet was supplemented with microalgae oil than with dry microalgae meal.     

A Saponification Method for Chlorophyll Removal from Microalgae Biomass as Oil Feedstock

Microalgae oil is an optimal feedstock for nutraceutical, pharmaceutical and biodiesel production, but its high levels of chlorophyll limit its large-scale application. To date, few effective approaches have been developed to remove chlorophyll from microalgae oil. The main purpose of this study was to present a preprocessing method of algae oil feedstock (Scenedesmus) to remove chlorophyll by saponification. The results showed that 96% of chlorophyll in biomass was removed. High quality orange transparent oil could be extracted from the chlorophyll reduced biomass. Specifically, the proportion of neutral lipids and saturation levels of fatty acids increased, and the pigments composition became carotenoids-based. The critical parameters of chlorophyll reduced biodiesel conformed to the standards of the USA, China and EU. Sodium copper chlorophyllin could be prepared from the bleaching effluent. The results presented herein offer a useful pathway to improve the quality of microalgae oil and reduce the cost of microalgae biodiesel.     

High-EPA Biomass from Nannochloropsis salina Cultivated in a Flat-Panel Photo-Bioreactor on a Process Water-Enriched Growth Medium

Nannochloropsis salina was grown on a mixture of standard growth media and pre-gasified industrial process water representing effluent from a local biogas plant. The study aimed to investigate the effects of enriched growth media and cultivation time on nutritional composition of Nannochloropsis salina biomass, with a focus on eicosapentaenoic acid (EPA). Variations in fatty acid composition, lipids, protein, amino acids, tocopherols and pigments were studied and results compared to algae cultivated on F/2 media as reference. Mixed growth media and process water enhanced the nutritional quality of Nannochloropsis salina in laboratory scale when compared to algae cultivated in standard F/2 medium. Data from laboratory scale translated to the large scale using a 4000 L flat panel photo-bioreactor system. The algae growth rate in winter conditions in Denmark was slow, but results revealed that large-scale cultivation of Nannochloropsis salina at these conditions could improve the nutritional properties such as EPA, tocopherol, protein and carotenoids compared to laboratory-scale cultivated microalgae. EPA reached 44.2% ± 2.30% of total fatty acids, and α-tocopherol reached 431 ± 28 µg/g of biomass dry weight after 21 days of cultivation. Variations in chemical compositions of Nannochloropsis salina were studied during the course of cultivation. Nannochloropsis salina can be presented as a good candidate for winter time cultivation in Denmark. The resulting biomass is a rich source of EPA and also a good source of protein (amino acids), tocopherols and carotenoids for potential use in aquaculture feed industry.     

piggyBac转座酶的瞬时表达对莱茵衣藻体内piggyBac转座子的作用

piggyBac转座系统作为一种遗传修饰的工具,在很多生物体如哺乳动物、昆虫、酵母中已经得到了广泛应用.然而,目前piggyBac转座系统在绿藻一莱茵衣藻(Chlamydomonas reinhardti)中是否有活性很少有入研究.本研究构建含有一个拷贝或多个拷贝并且能够稳定表达piggyBac转座元件的莱茵衣藻藻株.然后在含有转座元件的藻株中瞬时表达普通的piggyBac转座酶和密码子优化后的piggyBac转座酶,通过限制性片段长度多态性(restriction fragment length polymorphism,RFLP)分析piggyBac转座元件的转座现象.转座酶转入莱茵衣藻细胞内一周和两周之后与转入之前的RFLP图谱对比,有很明显的差异.转入转座酶一周后的TR1藻株原有的1.5kb大小的片段消失,同时出现了3条新的片段:2.8、4.2和6.5 kb.两周之后,这3条片段又发生了变化,表明转座现象存在并且持续发生.转入密码子优化后转座酶一周后的TR3藻株中,1.5、3和4kb大小的片段没有发生改变,但是6.5 kb大小的片段消失了,与此同时新出现了一个2.8 kb大小的片段.两周之后,TR3的限制性片段多态性图谱并没有发生变化.为了检测转座酶是否成功转入细胞中,我们在新的接头序列两侧进行侧翼PCR,同时对PCR产物进行序列比对分析,结果显示,piggyBac转座子偶尔会在非常规性位点5’-TTT-3’和5’-ACGCAG-3’处发生剪切,而常规性剪切位点发生在5’-TTAA-3’处.研究结果表明,piggyBac转座酶对莱茵衣藻体内piggyBac转座子具有转座作用,piggyBac转座系统可以用于莱茵衣藻的遗传学修饰.     

3种微藻兼养培养及营养成分的比较

对三角褐指藻Phaeodactylum tricormman、亚心形扁藻Platymonas subcordiformis、等鞭金藻Isochrysis galbana 3011品系分别进行自养培养和兼养培养试验,结果表明：三角褐指藻、等鞭金藻在兼养培养条件下,其生长速度都不同程度地增加,EPA和DHA含量也显著地提高;而亚心形扁藻在兼养培养条件下却表现出很大的差别,生长速度没有改变,EPA的含量极低,并且不含DHA。     

Replacing fish oil and astaxanthin by microalgal sources produced different metabolic responses in juvenile rainbow trout fed 2 types of practical diets

Dietary fish oil supplementation provides n-3 long-chained polyunsaturated fatty acids for supporting fish growth and metabolism and enriching fillet with eicosapentaenoic acid (EPA; 20:5n-3) and docosahexaenoic acid (DHA; c22:6n-3). Two experiments were performed as a 3 × 2 factorial arrangement of dietary treatments for 16 wk to determine effects and mechanisms of replacing 0%, 50%, and 100% fish oil with DHA-rich microalgae in combination with synthetic vs. microalgal source of astaxanthin in plant protein meal (PM)- or fishmeal (FM)- based diets for juvenile rainbow trout (Oncorhynchus mykiss). Fish (22 ± 0.26 g) were stocked at 17/tank and 3 tanks/diet. The 100% fish oil replacement impaired (P < 0.0001) growth performance, dietary protein and energy utilization, body indices, and tissue accumulation of DHA and EPA in both diet series. The impairments were associated (P < 0.05) with upregulation of hepatic gene expression related to growth (ghr1and igf1) and biosynthesis of DHA and EPA (fads6 and evol5) that was more dramatic in the FM than PM diet-fed fish, and more pronounced on tissue EPA than DHA concentrations. The source of astaxanthin exerted interaction effects with the fish oil replacement on several measures including muscle total cholesterol concentrations. In conclusion, replacing fish oil by the DHA-rich microalgae produced more negative metabolic responses than the substitution of synthetic astaxanthin by the microalgal source in juvenile rainbow trout fed 2 types of practical diets.