STUDIES ON MIXED MASS CULTIVATION OF ANABAENA SPP (NITROGEN-FIXING BLUE-GREEN-ALGAE, CYANOBACTERIA) ON A LARGE-SCALE

Studies on mixed mass cultivation of Anabaena spp. on a large scale (5170 m2) were conducted continuously for 3 years. Under the continental monsoon climate in northern subtropics (30-degrees-N, 115-degrees-E), 7-11 g dry weight m-2 day-1 of microalgal biomass on average was harvested in simple plastic greenhouses in the effective growth days during the warmer seasons. The maximum productivity was 22 g m-2 day-1 in the middle of summer. Observations on the productive properties of strains of Anabaena spp. indicated that they were different from and could compensate for each other in their productivities and adaptations to the seasonal changes. With different lining materials (PVC sheets, concrete, sand and soil) in the culture ponds, no significant variation of productivity was found, but bubbling with biogas in the middle of the day and the application of some growth regulating substances (2,4-D, NaHSO3 and extracts of oyster mushroom spawn) was able to improve the production. The cost of microalgal biomass in this way was around 0.75-1.0 US dollar(s) per kilogram.

Tank cultivation of the red alga Palmaria palmata: Year-round induction of tetrasporangia, tetraspore release in darkness and mass cultivation of vegetative thalli

Field-collected tetrasporophytes of Palmaria palmata were tumbled in 300-L outdoor tanks from January to August at ambient daylength or in a constant short-day (SD) regime (8 h light per day), both at 10 or 15 degrees C. Tetrasporangia were massively induced after 2.5 months under SD conditions at 10 degrees C and completely lacking at 15 degrees C, both under SD or ambient daylength conditions, with a few tetrasporangia present at 10 degrees C and ambient daylength. Elongation rates of tagged tetrasporophytic thalli peaked from March to April in all four conditions, when the biomass densities in the outdoor tanks were close to 2.5 kg fresh weight m(-2). Under all four conditions, juvenile proliferations started to appear in June from the margins of the old fronds, and attained approximately 1 cm in length by the end of July. Approximately 80% of the tetraspores were released during the first three dark phases in a light/dark regime, and the remaining 20% during the light phases. A minimum of 10 min darkness was observed to trigger spore release. White light inhibited tetraspore release, while a similar number of spores were released in continuous red light or in the light/dark regime, although with no significant differences of spore release during subjective days and nights. Sporelings were successfully derived from the released tetraspores for mass propagation of the male gametophyte in 2000-L outdoor tanks in a greenhouse. Mass production of male gametophytic sporelings of P. palmata was completed two times by SD induction of tetrasporangia at 10 degrees C, release of spores in darkness and culturing the sporelings until they were ready to be propagated vegetatively in greenhouse tanks. One experiment lasted from January to October 2001, with spore release in June, and the second from September to April 2003, with spore release in January. These results may support the development of sustainable, year-round Palmaria farming. (c) 2005 Elsevier B.V. All rights reserved.

Man-made desert algal crusts as affected by environmental factors in Inner Mongolia, China

Man-made desert algal crusts were constructed on a large scale (3000m(2)) in Inner Mongolia, China. Microcoleus vaginatus was mass cultivated and inoculated directly onto unconsolidated sand dune and irrigated by automatic sprinkling micro-irrigation facilities. The crusts were formed in a short time and could resist the erosion of winds and rainfalls 22 days after inoculation. The maximum biomass in the man-made algal crusts could also reach 35 mu g Chl a/cm(2) of soil. Effects of environmental factors such as temperature, irrigation, rainfall and soil nutrients on algal biomass of man-made algal crusts were also studied. It was found that rainfalls and lower light intensity had significantly positive effects on the biomass of man-made algal crusts. The preliminary results suggested that man-made algal crusts could be formed rapidly, and thus it might be a new feasible alternative method for fixing unconsolidated sand. (c) 2006 Elsevier Ltd. All rights reserved.

A simple and low-cost airlift photobioreactor for microalgal mass culture

A simple, low-cost, and efficient airlift photobioreactor for microalgal mass culture was designed and developed. The reactor was made of Plexiglas, and composed of three major parts: outer tube, draft tube and air duct. The fluid-dynamic characteristics of the airlift reactor were studied. The system proved to be well suited to the mass cultivation of a marine microalga, Chlorella sp. In batch culture, the biomass volumetric output rate of 0.21 g l(-1) d(-1) was obtained at the superficial gas velocity of 4 mm s(-1) in the draft tube.

Colony development and physiological characterization of the edible blue-green alga, Nostoc sphaeroides (Nostocaceae, Cyanophyta)

The edible blue-green alga, Nostoc sphaeroides Kutzing, is able to form microcolonies and spherical macrocolonies. It has been used as a potent herbal medicine and dietary supplement for centuries because of its nutraceutical and pharmacological benefits. However, limited information is available on the development of the spherical macrocolonies and the environmental factors that affect their structure. This report described the morphogenesis of N. sphaeroides from single trichomes to macrocolonies. During the process, most structural features of macrocolonies of various sizes were dense maculas, rings, the compact core and the formation of liquid core; and the. laments within the macrocolonies showed different lengths and arrays depending on the sizes of macrocolonies. Meanwhile temperature and light intensity also strongly affected the internal structure of macrocolonies. As microcolonies further increased in size to form 30 mm macrocolonies, the colonies differentiated into distinct outer, middle and inner layers. The. laments of the outer layer showed higher maximum photosynthetic rates, higher light saturation point, and higher photosynthetic effciency than those of the inner layer; whereas the. laments of the inner layer had a higher content of chlorophyll a and phycobiliproteins than those of the outer layer. The results obtained in this study were important for the mass cultivation of N. sphaeroides as a nutraceutical product. (c) 2008 National Natural Science Foundation of China and Chinese Academy of Sciences. Published by Elsevier Limited and Science in China Press. All rights reserved.

The past, present and future of phycology in China

Algae have been part of Chinese life for thousands of years. They are widely used as food and have been cited in Chinese literature as early as 2500 years ago. However, formal taxonomic studies on Chinese algae were initiated by foreign scientists only about 200 years ago, and by Chinese phycologists only about 90 years ago. This paper summarizes the history of modern phycological studies on Chinese algae and provides an overview of the achievements of phycological studies by Chinese scientists, especially on algal taxonomy, morphology, genetics, ecology and environmental research, physiology, biotechnology, algal culture, applied phycology and space phycology, in the last century. Recent development in phycological research focuses on algal floristic and molecular systematics, algal molecular biotechnology, applied phycology including micro and macroalgal cultivation and algal product development, and the roles of algae in environmental pollution control. These areas will also be the main focuses of Chinese phycological research in the foreseeable future.

Effect of iron on growth and lipid accumulation in Chlorella vulgaris

The economic feasibility of algal mass culture for biodiesel production is enhanced by the increase in biomass productivity and storage lipids. Effect of iron on growth and lipid accumulation in marine microalgae Chlorella vulgaris were investigated. In experiment I, supplementing the growth media with chelated FeCl3 in the late growth phase increased the final cell density but did not induce lipid accumulation in cells. In experiment II, cells in the late-exponential growth phase were collected by centrifugation and re-inoculated into new media supplemented with five levels of Fe3+ concentration. Total lipid content in cultures supplemented with 1.2 x 10(-5) mol L-1 FeCl3 was up to 56.6% biomass by dry weight and was 3-7-fold that in other media supplemented with lower iron concentration. Moreover, a simple and rapid method determining the lipid accumulation in C. vulgaris with spectrofluorimetry was developed. (c) 2007 Elsevier Ltd. All rights reserved.

Cultivation of the intertidal brown alga Hizikia fusiformis (Harvey) Okamura: mass production of zygote-derived seedlings under commercial cultivation conditions, a case study experience

Mariculture of the brown alga Hizikia fusiformis (Harvey) Okamura as an export-oriented human food has been there more for than 20 years in China. It is now one of the five major farmed algal species along the Chinese coast. Stable and sufficient supply of young seedlings for scaling up the cultivation has been a problem throughout the farming history of this species due to the unique dioecious life cycle and relatively short time window of sexual reproduction in nature. These two factors led to a practical difficulty in obtaining zygotes at identical developmental stage in viable amounts for seedling production. A key solution to this problem is to control the synchronization of the receptacle development and to realize the simultaneous discharge of male and female gametes, such that the fertilization rate could be greatly enhanced. Focusing on one of the farmed populations in this report, we present our results on mass production of seedlings using the synchronization technique on a large scale performed in 2007. Totally 5.5 hundred million embryos were obtained from 100 kg female sporophytes. The seedlings were raised up to 3.5 mm in length in greenhouse tanks over a month and were further grown in open sea for over 3 months at two experimental sites. The success of mass production of seedlings in this alga helped to lay the basis for future trials in other species in the genus of Sargassum that have identical life cycle.

Control of lunar and martian dust - Experimental insights from artificial and natural cyanobacterial and algal crusts in the desert of Inner Mongolia, China

Studies on the colonization of environmentally extreme ground surfaces were conducted in a Mars-like desert area of Inner Mongolia, People's Republic of China, with microalgae and cyanobacteria. We collected and mass-cultured cyanobacterial strains from these regions and investigated their ability to form desert crusts artificially. These crusts had the capacity to resist sand wind erosion after just 15 days of growth. Similar to the surface of some Chinese deserts, the surface of Mars is characterized by a layer of fine dust, which will challenge future human exploration activities, particularly in confined spaces that will include greenhouses and habitats. We discuss the use of such crusts for the local control of desert sands in enclosed spaces on Mars. These experiments suggest innovative new directions in the applied use of microbe-mineral interactions to advance the human exploration and settlement of space.

Chaetopyranin, a benzaldehyde derivative, and other related metabolites from Chaetomium globosum, an endophytic fungus derived from the marine red alga Polysiphonia urceolata

Cultivation of the endophytic fungus Chaetomium globosum, which was isolated from the inner tissue of the marine red alga Polysiphonia urceolata, resulted in the isolation of chaetopyranin (1), a new benzaldehyde secondary metabolite. Ten known compounds were also isolated, including two benzaldehyde congeners, 2-(2 ',3-epoxy-1 ',3 '-heptadienyl)-6-hydroxy- 5-(3-methyl-2-butenyl) benzaldehyde (2) and isotetrahydroauroglaucin (3), two anthraquinone derivatives, erythroglaucin (4) and parietin (5), five asperentin derivatives including asperentin ( 6, also known as cladosporin), 5 '-hydroxy-asperentin-8-methylether (7), asperentin-8-methyl ether (8), 4 '-hydroxyasperentin (9), and 5 '-hydroxyasperentin (10), and the prenylated diketopiperazine congener neoechinulin A (11). The structures of these compounds were determined on the basis of their spectroscopic data analysis (H-1, C-13, H-1-H-1 COSY, HMQC, and HMBC NMR, as well as low- and high-resolution mass experiments). To our knowledge, compound 1 represents the first example of a 2H-benzopyran derivative of marine algal-derived fungi as well as of the fungal genus Chaetomium. Each isolate was tested for its DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging property. Compounds 1-4 were found to have moderate activity. Chaetopyranin (1) also exhibited moderate to weak cytotoxic activity toward several tumor cell lines.

Cultivation of the brown alga Sargassum horneri: sexual reproduction and seedling production in tank culture under reduced solar irradiance in ambient temperature

As a large conspicuous intertidal brown alga, individuals of Sargassum horneri can reach a length of more than 7 m with a fresh weight of 3 kg along the coasts of the Eastern China Sea. The biomass of this alga as a vital component in coastal water ecology has been well documented. In recent years, a steady disappearance of the algal biomass along the once densely populated coastal areas of the Eastern China Sea has drawn attention in China. Efforts have been made to reconstruct the subtidal algal flora or even to grow the alga by use of long-lines. As part of the efforts to establish an efficient technique for producing seedlings of S. horneri, in this investigation a series of culture experiments were carried out in indoor raceway and rectangular tanks under reduced solar irradiance at ambient temperature in 2007-2008. The investigation demonstrated that: (1) sexual reproduction of S. horneri could be accelerated in elevated temperature and light climates, at least 3 months earlier than in the wild; (2) eggs of S. horneri had the potential to be fertilized up to 48 h, much longer than that of known related species; (3) suspension and fixed culture methods were both effective in growing the seedlings to the long-line cultivation stage; and (4) the life cycle of S. horneri in culture could be shortened to 4.5 months, thus establishing this alga as an appropriate model for investigating sexual reproduction in dieocious species of this genus.

Cultivation of the brown alga Hizikia fusiformis (Harvey) Okamura: enhanced seedling production in tumbled culture

The reuse of holdfasts for regeneration of young seedlings or using wild juvenile plants as the seedling source has played the major role in commercial cultivation of the brown alga Hizikia fusiformis in East Asia over the past 20 years. The possibility of employing zygote-derived germlings for producing seedlings has been discussed in the literature, but has not yet become a reality. Three main obstacles have limited the use of zygotes as a main source of seedlings, (1) the dioecious nature of the algal life cycle which may lead to asynchronous male and female receptacle development and thus different timing of egg and spermatozoa expulsion, (2) the low attachment rate when using zygote-derived germlings with developed rhizoids from wild parental plants for seeding production, and (3) the problem of culturing young germlings in regions where water temperature is high in summer. In this investigation, shifting the timing of receptacle formation earlier than in nature was performed by tumbling the algae in a long-day tank (16-h light per day). Synchronization of egg and spermatozoa expulsion and thereafter fertilization were conducted in indoor tanks. Receptacle formation in constant long days could be shifted by 20 days earlier than in plants cultured on long lines in the open sea, or I month earlier than in plants growing on intertidal rocks. Synchronized expulsion of eggs and spermatozoon led to a high rate of fertilization. This was achieved by tumbling the male and female receptacle-bearing branchlets in the same tank at low density in high irradiance. In two independent trials, a total of 1,400,000 zygote-derived germlings were obtained from 620 g (fresh weight) female sporophytes. The germlings shed from the receptacles were at an identical developmental stage indicating high synchronization of expulsion of eggs and spermatozoon followed by fertilization. Approximately 63% ( +/-9.6%) of the germlings were shed from the receptacle between 16 and 24 It after fertilization and 20% ( +/-11.9%) remained on the receptacle for 3 days after fertilization. Germlings were seeded on string collectors before rhizoids started to elongate and the attachment efficiency was enhanced. Young seedlings reached 800 ( +/-50) mum in length in 25 days at 25 degreesC before they were transferred to open sea cultivation. These results provide the basis of a practical way of seedling production by use of zygote-derived germlings in the commercial cultivation of Hizikia fusiformis. (C) 2004 Elsevier B.V All rights reserved.

Tank cultivation of the red alga Palmaria palmata: Effects of intermittent light on growth rate, yield and growth kinetics

Tank cultivation of marine macroalgae involves air-agitation of the algal biomass and intermittent light conditions, i.e. periodic, short light exposure of the thalli in the range of 10 s at the water surface followed by plunging to low light or darkness at the tank bottom and recirculation back to the surface in the range of 1-2 min. Open questions relate to effects of surface irradiance on growth rate and yield in such tumble cultures and the possibility of chronic photoinhibition in full sunlight. A specially constructed shallow-depth tank combined with a dark tank allowed fast circulation times of approximately 5 s, at a density of 4.2 kg fresh weight (FW) m(-2) s(-1). Growth rate and yield of the red alga Palmaria palmata increased over a wide range of irradiances, with no signs of chronic photoinhibition, up to a growth-saturating irradiance of approximately 1600 mumol m(-2) s(-1) in yellowish light supplied by a sodium high pressure lamp at 16 h light per day. Maximum growth rate ranged at 12% FW d(-1), and maximum yield at 609 g FW m(-2) d(-1). This shows that high growth rates of individual thalli may be reached in a dense tumble culture, if high surface irradiances and short circulation times are supplied. Another aspect of intermittent light relates to possible changes of basic growth kinetics, as compared to continuous light. For this purpose on-line measurements of growth rate were performed with a daily light reduction by 50% in light-dark cycles of 1, 2 or 3 min duration during the daily light period. Growth rates at 10degreesC and 50 mumol photon m(-2) s- 1 dropped in all three intermittent light regimes during both the main light and dark periods and reached with all three periodicities approximately 50% of the control, with no apparent changes in basic growth kinetics, as compared to continuous light.

Mass culture of Undaria gametophyte clones and their use in sporeling culture

Undaria pinnatifida (Harv.) Sur. is one of the three main seaweed species under commercial cultivation in China. In the mid-1990s the annual production was about 20000 tons dry. The supply of healthy sporelings is key to the success of commercial cultivation of Undaria. Previous studies demonstrated that instead of the zoospore collection method, sporelings can be cultured through the use of gametophyte clones. This paper reports the experimental results on mass culture of clones and sporeling raising in commercial scale. Light had an obvious effect on growth of gametophyte clones. Under an irradiance of 80 mumol m(-2) s(-1) and favorable temperature of 22-25degreesC, mean daily growth rate may reach as high as 37%. Several celled gametophyte fragments were sprayed onto the palm rope frame. Gametogenesis occurred after 4-6 days. Juvenile sporeling growth experiments showed that nitrate and phosphate concentrations of 2.9 10(-4) mol 1(-1) and 1.7 10(-5) mol 1(-1) were sufficient to enable the sporelings to maintain a high daily growth rate. Sporelings can reach a length of 1 cm in a month. Since 1997, extension of the clone technique has been carried out in Shandong Province. Large-scale production of sporelings for commercial cultivation of 14 and 31 hectares in 1997 and 1998 had been conducted successfully.

Algal biotechnology industries and research activities in China

In old China there were very few people engaged in the study of the algae, but in new China, freshwater and marine algae are studied by over one hundred old and new phycologists. There is now an algal biotechnology industry consisting of an aquaculture industry, producing large amounts of the seaweeds Laminaria, Porphyra, Undaria, Gracilaria, eucheumoids, and the microalgae Dunaliella and Spirulina. There is also a phycocolloid industry, producing algin, agar and carrageenan; an industry producing chemicals and drugs, such as iodine, mannitol, phycocyanin, beta -carotene, PSS (propylene glycol alginate sulfate) and FPS (fucose-containing sulfated polysaccharides) and an industry producing food, feed and fertilizer. The Laminaria cultivation industry produces about 900,000 t dry Laminaria, probably the largest producer in the world and 13,000 t algin, undoubtedly one of the largest algin producer in the world.