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The Second Algae Strain Report from AESI Published.

Hello friends,


   After a tremendous success from the first Algae strain report,AESI has published the second Algae Strain report.Our Institute and our sub-contractors Universities are getting success.


    Please read the report and mail me your view at scientistvivek01@gmail.com


You can get report from


  http://www.instituteofinnovations.com/projects.html


 

Tue May 24 2011 05:52:40 PM by Vivek 1

Algae Strain Report

Hello Friends,


      How are you?


Participating in the conferences in Italy,Japan & London, I am back here with report of some species which are being tested at Albert Einstein Science Institute,both in India & Germany..


  


    Description of the research performed at AESI in the area of microalgal collection and


screening. In addition to performing actual research in this area, AESI personnel were


responsible for coordinating the efforts of the many subcontractors performing similar activities,


and for standardizing certain procedures and analyses. These efforts ultimately resulted in the


development of the AESI Microalgal Culture Collection, which is more fully described in


Chapter II.A.3.


Brief review of algal taxonomic groups and characteristics:


For the purposes of this report, microalgae are defined as microscopic organisms that can grow


via photosynthesis. Many microalgae grow quite rapidly, and are considerably more productive


than land plants and macroalgae (seaweed). Microalgae reproduction occurs primarily by


vegetative (asexual) cell division, although sexual reproduction can occur in many species under


appropriate growth conditions.


There are several main groups of microalgae, which differ primarily in pigment composition,


biochemical constituents, ultrastructure, and life cycle. Five groups were of primary importance


to the ABP: diatoms (Class Bacillariophyceae), green algae (Class Chlorophyceae), goldenbrown


algae (Class Chrysophyceae), prymnesiophytes (Class Prymnesiophyceae), and the


eustigmatophytes (Class Eustigmatophyceae). The blue-green algae, or cyanobacteria (Class


Cyanophyceae), were also represented in some of the collections. A brief description of these


algal groups follows.


? Diatoms. -Diatoms are among the most common and widely distributed groups


of algae in existence; about 100,000 species are known. This group tends to


dominate the phytoplankton of the oceans, but is commonly found in fresh- and


brackish-water habitats as well. The cells are golden-brown because of the


presence of high levels of fucoxanthin, a photosynthetic accessory pigment.


Several other xanthophylls are present at lower levels, as well as β-carotene,


chlorophyll a and chlorophyll c. The main storage compounds of diatoms are


lipids (TAGs) and a β-1,3-linked carbohydrate known as chrysolaminarin. A


distinguishing feature of diatoms is the presence of a cell wall that contains


substantial quantities of polymerized Si. This has implications for media costs


in a commercial production facility, because silicate is a relatively expensive


chemical. On the other hand, Si deficiency is known to promote storage lipid


accumulation in diatoms, and thus could provide a controllable means to induce


lipid synthesis in a two-stage production process. Another characteristic of


diatoms that distinguishes them from most other algal groups is that they are


diploid (having two copies of each chromosome) during vegetative growth;


most algae are haploid (with one copy of each chromosome) except for brief


periods when the cells are reproducing sexually. The main ramification of this


from a strain development perspective is that it makes producing improved


strains via classical mutagenesis and selection/screening substantially more


difficult. As a consequence, diatom strain development programs must rely


heavily on genetic engineering approaches.


? Green Algae-. Green algae, often referred to as chlorophytes, are also abundant;


component. However, N-deficiency promotes the accumulation of lipids in


certain species. Green algae are the evolutionary progenitors of higher plants,


and, as such, have received more attention than other groups of algae. A


member of this group, Chlamydomonas reinhardtii (and closely related species)


has been studied very extensively, in part because of its ability to control sexual


reproduction, thus allowing detailed genetic analysis. Indeed, Chlamydomonas


was the first alga to be genetically transformed. However, it does not


accumulate lipids, and thus was not considered for use in the ABP. Another


common genus that has been studied fairly extensively is Chlorella.


? Golden-Brown Algae-. This group of algae, commonly referred to as


chrysophytes, is similar to diatoms with respect to pigments and biochemical


composition. Approximately 1,000 species are known, which are found


primarily in freshwater habitats. Lipids and chrysolaminarin are considered to


be the major carbon storage form in this group. Some chysophytes have lightly


silicified cell walls.


? Prymnesiophytes. -This group of algae, also known as the haptophytes, consists


of approximately 500 species. They are primarily marine organisms, and can


account for a substantial proportion of the primary productivity of tropical


oceans. As with the diatoms and chrysophytes, fucoxanthin imparts a brown


color to the cells, and lipids and chrysolaminarin are the major storage products.


This group includes the coccolithophorids, which are distinguished by


calcareous scales surrounding the cell wall.


? Eustigmatophytes-. This group represents an important component of the


?picoplankton?, which are very small cells (2-4 μm in diameter). The genus


Nannochloropsis is one of the few marine species in this class, and is common


in the world?s oceans. Chlorophyll a is the only chlorophyll present in the cells,


although several xanthophylls serve as accessory photosynthetic pigments.           


 


*Cyanobacteria-. This group is prokaryotic, and therefore very different from all


other groups of microalgae. They contain no nucleus, no chloroplasts, and have


a different gene structure. There are approximately 2,000 species of


cyanobacteria, which occur in many habitats. Although this group is


distinguished by having members that can assimilate atmospheric N (thus


eliminating the need to provide fixed N to the cells), no member of this class


produces significant quantities of storage lipid; therefore, this group was not


deemed useful to the ABP.



Growth rates.


Six promising strains were analyzed in AESI Type I, Type II, and ASW (Rila) using the


temperature-salinity gradient described previously. These included the diatoms Chaetoceros


muelleri , Navicula , Cyclotella , Amphora 1 & 2, and the chlorophyte Monoraphidium minutum . Navicula and


Cyclotella were actually collected from the Florida keys; the remaining strains were collected in


Colorado and Utah.) All strains exhibited rapid growth over a wide range of conductivities in at


least two media types. Furthermore, all strains exhibited temperature optima of 30?C or higher.


Maximal growth rates of these strains, along with the optimal temperature, conductivity, and


media type determined in these experiments are shown in Table II.A.1. (Higher growth rates     


were determined for some of these strains in subsequent experiments.


       Experiments were also conducted in an attempt to identify the chemical components of AESI


Type I and Type II media most important for controlling the growth of the various algal strains.


Bicarbonate and divalent cation concentrations were found to be important determinants in


controlling the growth of Boekelovia sp. and Monoraphidium . The


growth rate of Monoraphidium minutum 2 .  increased by more than five-fold as the bicarbonate concentration of


Type II/25 medium was increased from 2 to 30 mM, and the growth of BOEKE1 by


approximately 60% over this range. These results make sense, since media enriched in


bicarbonate would have more dissolved carbon available for photosynthesis. An unexpected


finding was that there was a decrease of nearly 50% in the growth rate of BOEKE1 as the


divalent cation concentration increased from 5 mM to 95 mM (in Type I/10 medium containing


altered amounts of calcium and magnesium). The effects of magnesium and calcium


concentration on the growth of Monoraphidium minutum .  were less pronounced. These results indicate that


matching the chosen strain for a particular production site to the type of water available for mass


cultivation will be important.


Lipid content.


The lipid contents of several strains were determined for cultures in exponential growth phase


and for cultures that were N-limited for 7 days or Si-limited for 2 days. In general, nutrient


deficiency led to an increase in the lipid content of the cells, but this was not always the case.


The highest lipid content occurred with Navicula 1, which increased from 22% in exponential


phase cells to 49% in Si-deficient cells and to 58% in N-deficient cells. For the green alga


Monoraphidium minutum ., the lipid content increased from 22% in exponentially growing cells to 52% for cells


that had been N-starved for 7 days. Chaetoceros


muelleri also exhibited a large increase in lipid content in


response to Si and N deficiency, increasing from 19% to 39% and 38%, respectively. A more


modest increase occurred for nutrient-deficient AMPHO1 cells, whereas the lipid content of


Cyclotella was similar in exponential phase and nutrient-deficient cells, and actually decreased in


Amphora 2  as a result of nutrient deficiency.


    These results suggested that high lipid content was indeed achievable in many strains by


manipulating the nutrient levels in the growth media. However, these experiments did not


provide information on actual lipid productivity in the cultures, which is the more important


factor for developing a commercially viable biodiesel production process. This lack of lipid


productivity data also occurred with most of the ABP subcontractors involved in strain screening


and characterization, but was understandable because the process for maximizing lipid yields


from microalgae grown in mass culture never was optimized. Therefore, there was no basis for


designing experiments to estimate lipid productivity potential.


 


Six algal  Strains


 


 (B. braunii, Dunaliella primolecta, Isochrysis sp., Monallanthus salina, Phaeodactylum


tricornutum, and Tetraselmis sueica) were obtained from existing culture collections and


analyzed with respect to lipid, protein, and carbohydrate content under various growth


conditions. For these experiments, all cultures except for B. braunii were grown in natural


seawater that was enriched with N, P, and trace metals. B. braunii was grown in an artificial


seawater medium. Initial experiments to determine productivities of these species were


performed using batch cultures in 9-L serum bottles. Of the strains tested, the highest growth


rates were observed with P. tricornutum (Thomas strain) and M. salina.


Additional experiments were performed in plexiglas vessels that were 5 cm thick, 39 cm deep,


2,000-watt tungsten-halide lamp, which was placed behind a water/CuSO4 thermal filter. In


these experiments, the cultures were typically maintained for 40 to 90 days. In the early stages of


an experiment, the cultures were maintained in a batch mode, and then converted to a continuous


or semi-continuous dilution mode. Various culture parameters (including light intensity, dilution


rate, and N status) were manipulated during the course of these experiments to determine their


effects on the productivities and proximate chemical composition of the strains. The results of


these experiments with each species tested are discussed below. These experiments are difficult


to compare because the experiments were all carried out slightly differently (i.e., different light


intensities, different culturing methods [batch, semi-continuous, and continuous], different means


of obtaining N-deficient cultures, and inconsistent use of a CuSO4 heat filter, which resulted in


differences in light quality and culture temperature). Nonetheless, the general conclusions of this


study are of interest.


P. tricornutum):


This strain has been used for several past studies, and was concomitantly being tested in outdoor


mass culture by another subcontractor (the University of Hawaii; principal investigator Dr.


Edward Laws; discussed in Section III). Therefore, this strain was subjected to more extensive


testing than the other strains in this subcontract. In one experiment reported for this strain, the


effects of light intensity on productivity were determined in batch cultures (i.e., in the Plexiglas


culture apparatus described earlier without culture replacement and dilution). The maximum


productivity observed for this strain (21 to 22 g dry weight?m-2?d-1)3 was observed at a total daily


illumination of 63-95 kcal (representing approximately 40%-60% of full sunlight in southern


California during the summer). This value was slightly higher than the productivity observed


with a total daily illumination of 70% full sunlight (17.1 g dry weight?m-2?d-1). Productivities


under N-limiting, continuous growth mode conditions were between 7 and 11 g dry


weight?m-2?d-1. Likewise, productivities under N-sufficient, continuous growth mode conditions


were reduced relative to batch cultures.


In addition to measuring overall productivities, the levels of protein, carbohydrate, lipid, and ash


were determined for cells grown under the various conditions described earlier. Illumination of


the cultures from 40% to 70% of full sunlight did not have a large impact on the cellular


composition. Growth of P. tricornutum cells under N-deficient conditions resulted in a reduction


of the protein content from 55% (in N-sufficient cells) to 25% of the cellular dry weight.


Carbohydrate content increased from 10.5% to 15.1%, and the mean lipid content increased from


19.8% to 22.2%, although these differences in carbohydrate and lipid contents did not appear to


be statistically significant. At one stage of the experiment, however, a time course of N


deficiency led to a consistent rise in lipid content from 19.9% to 30.8% over the course of 7 days.


The actual rate of lipid production did not increase, however, because the overall productivity of


the cultures was reduced under N-deficient growth.


D. primolecta:


The maximum productivity observed for this species (12.0 g dry weight?m-2?d-1 ) occurred during


continuous culture at 60% full sunlight under N-sufficient conditions. Doubling the light


intensity lowered the productivity to 6.1 g dry weight?m-2?d-1. The chemical composition of Nsufficient


cells (as an average percentage of total cell dry weight) was 64.2% protein, 12.6%


carbohydrate, and 23.1% lipid. After 7 days of growth under N-deficient conditions, the


composition was 26.8% protein, 59.7% carbohydrate, and 13.7% lipid. Therefore, this alga


accumulates carbohydrates rather than lipids in response to nutrient deficiency, limiting its


usefulness as a lipid production strain.


M. salina:


This alga reportedly contained high levels of lipids when grown under N-deficient conditions.


The highest productivity (13.9 g dry weight?m-2?d-1) was observed under N-sufficient conditions


at a light intensity of 50% full sunlight, although detailed experiments with regards to the effects


of light intensity on productivity were not conducted. There was little difference in the lipid


3Reporting of productivities in g dry weight?m-2 ?d-1 derives from the goal of mass culturing the algae in


shallow open ponds. The objective would be to maximize biomass produced per area of pond. However, it


is often difficult to compare results between experiments when the data are reported in this manner, as


factors such as culture depth and vessel design would significantly affect productivity of the cultures.


content of cells grown under N-sufficient and N-deficient conditions (20.7% and 22.1%,


respectively).


 


 


T. sueica:


The highest productivity observed for this strain was 19.1 g dry weight?m-2?d-1, which occurred


in N-sufficient batch cultures grown under a light intensity of 60% full sunlight. N deficiency


resulted in a large increase in carbohydrate content (from a mean value of 10.7% to a mean value


of 47.1%). On the other hand, protein content was reduced substantially (from 67.6% to 28.3%),


and the lipid content decreased from 23.1% to 14.6% in response to N deficiency.


Isochrysis sp.(Tahitian strain T-ISO):


This strain is commonly used as a feed organism in aquaculture production systems. A


productivity of 11.5 g dry weight?m-2?d-1 was typical for batch cultures of this species, which was


approximately 33% higher than the value recorded during semi-continuous growth (dilution of


0.15 L/d). Productivity was lowered during N-deficient growth to 5.5-7.6 g dry weight?m-2?d-1.


This strain accumulated carbohydrate in response to N deficiency (from a mean value of 23.1%


to 56.9%). Lipid content also increased slightly (from 28.5% to 33.4%), whereas protein content


was reduced from 44.9% to 27.3%. The higher lipid content of N-deficient cells did not translate


to higher lipid productivities, however, because of the lower overall productivity of the stressed


cultures.


B. braunii:


Some very limited experiments were conducted with this species, which is known to accumulate


hydrocarbons. A culture grown under a light intensity of 60% full sunlight had a productivity of


only 3.4 g dry weight?m-2?d-1. The lipid content of these cells was 29% of the cellular dry


weight; the N status of the cells was not reported, but it is assumed that the cells were grown


under N-sufficient conditions.


Overall Conclusions


Of the species examined, P. tricornutum and T. sueica had the highest overall productivities.


These species also had the highest lipid productivities, which were 4.34 and 4.47 g lipid?m-2?d-1,


respectively. For both species, the maximal productivities were obtained in batch cultures, as


opposed to semi-continuous or continuous cultures. Although the lipid contents of cells were


often higher in response to N deficiency, the lipid productivities of all species tested were


invariably lower under N deficiency because of an overall reduction in the culture growth rates.


For the species tested under continuous or semi-continuous growth conditions, lipid


productivities were reduced from 14% to 45% of the values measured for N-sufficient cultures.


The results also pointed to the importance of identifying strains that are not photoinhibited at


light intensities that would occur in outdoor ponds. Finally, this work highlighted the fact that


some microalgae accumulate carbohydrates during nutrient-deficient growth; such strains are


clearly not acceptable for use as a feedstock for lipid-based fuel production.


    The work is going on....


Will share you more when the reports will come..


 Please give your views..


 



 

Thu November 04 2010 09:50:50 AM by Vivek 9

Greatest Biodiesel Extraction Can Be Done From Chlorophyceae Algaes

The Chlorophyceae are a large and important group of freshwater green algae. They include some of the most common species, as well as many members that are important both ecologically and scientifically.
There are approximately 350 genera and 2650 living species of chlorophyceans. They come in a wide variety of shapes and forms, including free-swimming unicellular species, colonies, non-flagellate unicells, filaments, and more. They also reproduce in a variety of ways, though all have a haploid life-cycle, in which only the zygote cell is diploid. The zygote will often serve as a resting spore, able to lie dormant though potentially damaging environmental changes such as dessication.
The Chlorophyceae includes three major groups distinguished primarily by basic differences in the arrangement of their flagellae:
? Volvocales, Chaetophorales, & Chlorococcales - together make up more than half of all chlorophyceans. Members of these orders have an offset flagellar arrangement (1 o'clock-7 o'clock).
? Chlorellales - Members of this order have opposed flagellae (12 o'clock-6 o'clock), though some have only vestigial flagellae and so have not been definitively associated with this group. Similarities with members of the Chlorococcales make distinctions difficult.
? Oedogoniales - Members of this smallest group have a complex multiflagellate crown on their swimming spores. All are filamentous, oogamous, and have net-like chloroplasts.
Chlorophycean Diversity : Above are shown representatives of the three major groups of Chlorophyceae. From left to right, these are Chlamydomonas (Volvocales); Pediastrum (Chlorellales); and Oedogonium (Oedogoniales). The pictures also serve to show some of the structural diversity of chlorophyceans; these are (again, from left to right): unicells, colonies, and filaments.

Perhaps the most famous and important chlorophyceans are Chlamydomonas and Volvox. Both are important research organisms in biological laboratories. Volvox, pictured at the top of this page, is a model organism for understanding cell death and cell interactions. Chlamydomonas, pictured above at left, though tiny and single-celled has been useful in countless studies, including freshwater ecology, cell division, physiology, genetic structure and function, and genomic evolution.
No less important are the Chlorellales, especially Chlorella. Though only about ten species are known in this genus, it plays an important role as endosymbionts inside the tissues of other organisms. Sponges, polyps, ciliates, and forams all may house Chlorella internally, providing a home for the alga in exchange for its photosynthates. More recently, this tiny alga has become popular as a "health food" for its concentrated proteins and chlorophyll, and is now raised commercially in large numbers, especially in Asia.
Many other species of Chlorophyceae are common. Oedogonium grows in freshwater worldwide, usually attached to other plants or algae; it has been a subject of intense study for its unusual cell division method. Species of Chlorococcales are frequently found in soil or mud, and some, such as Apatococcus may be found growing in bright green patches on walls and the sides of trees where moist conditions prevail.

The oldest fossils resembling Chlorophyceans come from the Proterozoic Bitter Springs Formation of central Australia. There is insufficient preservation to make a firm identification of these fossils, but other multicellular algae are known from this time (Butterfield et al. 1988), so the identification is not unreasonable.
Additionally fossils of this group have been found in Middle Devonian rocks from New York State (Baschnagel 1966). Among these is Paleooedogonium, a fossil with striking resemblance to modern Oedogonium. The presence of this fossil and those of other freshwater green algae in the Devonian suggests that the group was quite diverse by this time.
Tertiary fossils of modern chlorophycean genera are also known, such as Scenedesmus in the Chlorellales (Fleming 1989), Pediastrum and Botryococcus (Gray 1960), and Tetraedon (Goth et al. 1988). The last of these is believed to be an important component in the formation of oil shales of the Tertiary, such as those in Messel, Germany.

In my view the greatest oil extraction can be done from this algae.
Please give your views.
Sun August 15 2010 10:47:21 AM by Vivek 8