Oil recovery from wet Euglena gracilis by shaking with liquefied dimethyl ether

https://www.researchgate.net/publication/297890244_Oil_recovery_from_wet_Euglena_gracilis_by_shaking_with_liquefied_dimethyl_ether

Algal biomass gasification efficiencies

Suffice to say i have had cross contaminations of euglena species throughout all of my samples so far.

I suspect that is has somehow travelled through my single air/co2 pump system and into the isolated cultures.

At this point it would be easy to say that euglena is an enemy/parasite of my algaculture endeavors.

But sometimes it is necessary to take a step back and ask. IS IT AND ENEMY or is it just more prolific?

It would be easy to just state it us an enemy or parasite and 'attempt' to take additional measures to eliminate euglena from the samples. But that is MORE TIME AND MORE EXPENSE to try to eliminate a USEFULL SPECIES.

I have been doing extensive studies ans a myriad of thought experiments on how to keep euglena out of the samples. Unless on has a clean roon type of laboratory at hand and extensive protocols in place and of which can be continuously inplemented. ITNOS VERY EXPENSIVE.

However, on the other hand. After aeeing the success of my fathers development of a gasification of wood chips ( a different for of biomatter ) to produce hydrogen rich syngas capable of turning a 10,000 watt electric generator. I have been researching the feasibility of using dry algae for gasification syngas generation and have found that it is not only feasible but VERY Efficient.

This conclusion is that using natural PROLIFIC INVASIVE species of algae or euglena in an open pond /  raceway environment to WORK WITH NATURE to produce euglena/ algal biomass to more efficiently produce biomass for gasification instead of focusing on producing algae OIL for biodiesel production using unnatural, sterile an expensive laboratory environment is more efficient and more natural.

So my focus then ia to focus on producing as much algal biomass as possible and not focus so much on OIL for BIODIESEL production which, from my research,  is inefficient and in exorbitantly costly.

AND ALSO IN CREATING A SMALL SCALE ALGAE GASIFICATION SYNGAS GASIFIER.

Gasification of algae biomass

http://www.sciencedirect.com/science/article/pii/S092058611400409X

Intriguing article- Production of Biorenewable Hydrogen and Syngas via Algae Gasification: A Sensitivity Analysis

http://www.sciencedirect.com/science/article/pii/S1876610214033311

Enhancing Algal Growth by Stimulation with LED Lighting and Ultrasound

Enhancing Algal Growth by Stimulation with LED Lighting and Ultrasound

https://www.hindawi.com/journals/jnm/2015/531352/

Very cool Instructables on an algae photobioreactor called the BIOMONSTAAR

PRETTY COOL SETUP


http://www.instructables.com/id/Biomonstaaar/

THIS INSTRUCTABLE GOES INTO HOW TO CREATE THE SENSORS WITH ARDUINO.

http://www.instructables.com/id/Arduino-system-for-Biomonstaaar-bioreactor/

Three Dollar EC - PPM Meter [Arduino]

https://hackaday.io/project/7008-fly-wars-a-hackers-solution-to-world-hunger/log/24646-three-dollar-ec-ppm-meter-arduino

Interesting information on the vapor point of water under vacuum.

Waters boiling point in a vacuum

http://www.engineersedge.com/h2o_boil_pressure.htm

intresting 5 gallon water bottle - photobioreactor design

I happen to have one of these laying around and am thinking of creating a 5 gallon pbr to grow my euglena in???

Free pdf from http://off-grid-living.com/- http://off-grid-living.com/wp-content/uploads/2016/05/Algae-Biofuels-sample2.pdf

will research this more..

More research on Euglena - can it be used for Biofuel production??

Since my Euglena seem to be super invasive across most of my samples ( most likely coming from the air supply tubing ) is it possible to grow euglena as a biofuel?

here is an informative  article on Encyclopedia Britannica - https://www.britannica.com/science/Euglena


Euglena are characterized by an elongated cell (15–500 micrometres [1 micrometre = 10−6 metre], or 0.0006–0.02 inch) with one nucleus, numerous chlorophyll-containing chloroplasts (cell organelles that are the site of photosynthesis), a contractile vacuole (organelle that regulates the cytoplasm), an eyespot, and one or two flagella. Certain species (e.g., E. rubra) appear red in sunlight because they contain a large amount of carotenoid pigments. Unlike plant cells, Euglena lack a rigid cellulose wall and have a flexible pellicle (envelope) that allows them to change shape. Though they are photosynthetic, most species can also feed heterotrophically (on other organisms) and absorb food directly through the cell surface via phagocytosis (in which the cell membrane entraps food particles in a vacuole for digestion). Food is often stored as a specialized complex carbohydrate known as paramylon, which enables the organisms to survive in low-light conditions. Euglena reproduce asexually by means of longitudinal cell division, in which they divide down their length, and several species produce dormant cysts that can withstand drying.

http://link.springer.com/article/10.1007/s10811-013-9979-5

Euglenoids achieved a maximum lipid content of 24.6 % (w/w) with a biomass density of 1.24 g  L−1 (dry wt.).


The japanese company - apply named EUGLENA - states - 

Euglena's oil formation is suited for jet fuels

A jet engine uses kerosene, which is lighter than diesel but heavier than gasoline. The oil extracted and refined from euglena happens to be well suited to be refined in to kerosene compared to other algal oils.

High productivity and efficient land use

Since euglena uses special equipment for cultivation, it does not compete over farmland with food unlike biofuels derived from corn and sugarcane.
Productivity per acre is also significantly higher compared to many other feedstocks.
In our laboratory in Tokyo University, we have achieved 15 times higher productivity compared to palm oil.

Alternative fuel

Fossil fuels releases CO2 captured underground and releases it into the atmosphere, causing global warming.
On the other hand euglena absorbs CO2 from the atmosphere during production so there is a chance of reducing CO2 emission
if there are further technical developments.

http://biofuels-news.com/ has another article on euglena - DECEMBER 2, 2015 - Japanese Euglena to establish an algae-based jet fuel demo plant

Euglena, a Japanese microalgae developer and producer, has partnered with Japan’s largest airline ANA Holdings to develop an algae-based jet fuel.

A 3 billion yen (appr. €23m) demonstration plant, using technology from Chevron, is scheduled to come online in early 2018 with plans to 125,000 litres of renewable jet fuel.

Euglena hopes to be able to commercialise the fuel, derived from its namesake euglena algae, by 2020 and set up more facilities with output more than 400 times that of the demonstration plant.

The company also want to set up large-scale cultivation facilities outside of Japan.

The algae fuel, refined in the US from oil extracted from euglena, has a chemical makeup similar to the of jet kerosene.

‘We'll use the fuel from the demonstration plant on real flights, mixing it with standard oil-based fuel,’ says Kiyoshi Tonomoto, executive VP at ANA Holdings.

The International Civil Aviation Organisation aims to stabilise the aerospace industry’s CO₂emissions at 2020 levels.

Biofuels are attracting attention as a carbon-neutral option to help achieve this goal, since plants absorb carbon dioxide as they grow and release it when burned.

The biggest hurdle to commercializing the algae-derived fuel will be reducing the price, which is reportedly ten times higher than petroleum-derived kerosene.

To me it would seems that one would chose the most prolific species of algae, in my samples it is totally invasive and takes over all my samples, and use it instead of trying to control all of the environmental parameters to try too keep euglena out.

One just needs to research and find the correct parameters to make FAT EUGLENA??

Another article - The Potential in Your Euglena Pond - 

http://advancedbiofuelsusa.info/the-potential-in-your-euglena-pond/

(Algae Industry Magazine)  Scientists at the John Innes Centre, in Norwich, England, have discovered that Euglena gracilis, the single cell algae that inhabits most garden ponds, has a whole host of new, unclassified genes that can make new forms of carbohydrates and natural products.

Even with the latest technologies, sequencing the DNA in Euglena remains a complex and longwinded undertaking. Dr. Ellis O’Neill and Professor Rob Field from the John Innes Centre have therefore sequenced the transcriptome of Euglena gracilis, which provides information about all of the genes that the organism is actively using.

The team also found that different sets of genes become active when Euglena is grown in the dark to when it is grown in the light. This indicates that Euglena can dramatically shift its metabolism depending on its environment, which reflects its ability to live successfully in varied environments.


Euglena creates many well-known, valuable natural products including vitamins, essential amino acids and a sugar polymer, which is reported to have anti-HIV effects. Given the usefulness of the compounds we know about, these findings have the potential, with further research, to lead to the discovery of new medicines including new antibiotics, nutrients and new forms of biofuel, among other products.

Biofuel from Euglena

09 March 2010

JAPAN - Japanese oil company, Nippon Oil and Hitachi subsidiary Hitachi Plant Technologies have joined forces to develop jet fuel from a single celled pond and lake organism, Euglena.

The companies have acquired shared in Euglena Inc, which is developing systems to extract oil from the algae to produce fuel.

Euglena, meaning “beautiful eye”, is a single-celled algae that can be found in lakes, ponds, and even puddles. It was named after Leeuwenhoek, a microscope inventor in the 17th century, who incidentally found Euglena through the lens. Euglena has the characteristics of both animal and plant, where it moves around like an animal, and photosynthesises like a plant.

The joint venture has a culturing system in place that can be used to grow Euglena efficiently and the yield is better than crops such as corn and sugar cane usually used to produce biofuel, a report in Crunch Gear says.

The company is trying to lower the cost of production to 80 cents a litre to make biofuel production competitive.

The company says mass-producing Euglena-derived biofuel should be possible by 2015.

SC11192016 Stoners creek growth - 2 weeks - more euglena!!!

It took this sample a while to produce - EUGLENA!!! and a small amount of chlorella.

this sample not producing very well - especially since I believe my current air supply setup is contaminating my samples.

2ND FROM LEFT

FY11012016 Front Yard Algae Culture Upscale - results 12-03-2016

Almost completely all euglena!!! definately the dominate species here.

this sample is from the upscale done on 11/21/2016 - 12 days and ( what I am calling euglena )  has taken over completely!!

moody ramp ( runnoff of Dale hollow lake dam ) - filamentous algae and euglena and ameobic invasive patterns

The Moody Ramp sample taken 11-11-2016 - showing signs of euglena invasion and long chains of filamentous algae species     - also what appears to be amoebic activity

Euglena invading algae sample - Percy priest Lake sample - PPL11102016 video

Euglena invading sample - AGAIN. This seems to be across the board on all my samples - starting to suspect unfiltered air supply...

These videos show a continuation of an invasion of euglena??
This sample clearly shows euglena invasion and almost wiped out other species....

reference -
http://www.landcareresearch.co.nz/resources/identification/algae/identification-guide/identify/guide/unicellular/euglena

in this video you can clearly see the cells changing shape -

Chlorella mononculture!!! LCL11112016

After a couple of days of just letting the system set and grow I have significant grow in all samples.

On today's first sample microscopic inspection I decided to test the LCL11112016 sample taken from a local city lake and was surprised at what I found!!

Almost complete dominance of Chlorella type species with a few super fast swimming parasites feeding on the sample. Also saw several different stages of growth from full grown to dividing to colony structures.

A "Good Enough" Stage Micrometer

http://www.microscopy-uk.org.uk/mag/indexmag.html?http://www.microscopy-uk.org.uk/mag/artoct02/hwmicrometer.html

Life cycles algae & plants

Copied from - http://www.vcbio.science.ru.nl/en/virtuallessons/lifecycle/

Life cycles algae & plants

Organisms, from little moss to man, go during their life through a number of biological phases, generation after generation. The following "biological phases" are encountered during the life cycle: the formation of gametes (the sexual reproductive cells), the fusion of male and female gametes (~ fertilization) to a zygote, and a period of growth and development (cell differentiation and morphogenesis) that occurs at various time sequences, depending on the phylogenetic group to which the organism belongs. Within the life cycle, meiotic divisions (more) take place, in which cell proceed from a diploid to a haploid stage; in the diploid stage two homologous copies of each chromosome are present in the cells and in the haploid stage only one sample of each chromosome in present in each cell. Gametes are always haploid, whereas zygotes are diploid. Furthermore, during the life cycle mitotic divisions occur (more about mitosis). Depending on the phylum to which the organism belongs, these mitotic divisions occur in the haploid, or diploid phase, or in both the haploid and diploid stages. More here about here below.  In these web modules one finds descriptions of the life cycles of brown, red and green algae and that of various phyla of land plants. The characteristics are shown according to typical representants of each group, the same as those that are discussed in the course "Evolution and Development of the Plant" (Radboud University Nijmegen). The classification and terminology in these webpages follows the handbook that is used during lectures and practical courses: "Biology, Campbell, Reece, Urry, Cain, Wasserman, Minorsky en Jackson, 2008. Pearson International Edition (eds).San Francisco. 1267 pp".    Classification of brown, red, green algae and land plants within the Eukaryotes Phyla that are discussed in this module are indicated by a red dot Three types of life cycles can be distinguished on account of the timing of the mitoses, in the haploid and/or diploid phase: Diplontic: In diplonts mitotic divisions only occur in diploid cells. Gametes (arisen through meiosis) are the only occurance of the haploid phase. The diploid zygote formed after fertilization can however divide mitotically. This process results in the production of a multicellular diploid organism or in the production of many diploid single cells. Animals, for example, belong to the diplonts.   Haplontic: In haplonts the mitoses only occur in haploid cells. This can result in the formation of single haploid cells or a multicellular haploid organism. The haplontic life forms produce the gametes through mitosis. After fertilization a zygote is formed: this cell is the only diploid cell in the entire life cycle. It is thus that same zygotic cell that later undergoes meiosis. A haploid life cycle is found in most fungi and in some green algae like Chlamydomonas.   Haplo-diplontic:  In haplo-diplonts the mitoses occur in both diploid and haploid cells. Such organims go during their life cycle through a phase in which they are multicellular and haploid (the gametophyte), and a phase in which they are multicellular and diploid (the sporophyte). The phenomenon is called "alternation of generation". This haplo-diplontic type of cycle is found in all land plants and in many algae.   Colofon This series webpages on the life cycle in plants is based on course notes and other teaching material of Mieke Wolters-Arts, Celestina Mariani, Jan Derksen, the late G. van den Ende and Luud J.W. Gilissen. Dr Gerard van den Ende had developed an extended manual enriched with numerous drawings of life cycles, which went along with the practical courses that he taught. The microscopical work and the layout are by Liesbeth Pierson (contact), Wilbert Janssen and Ines Schulten. The web structure was developed by Remco Aalbers.

Inoculation of 50ml growth culture take from petri dish cultures into 500ml growth media.

Inoculation of 50 ml growth culture take from petri dish cultures into 500 ml growth media.

After 5 days growth of media taken from the front yard growth media petri dish sample and inoculate a 500 ml growth media.

Before this I examined the sample under microscope and found nothing but what I am labeling Characium pringsheimii based upon visual comparison.

very distinctive structure

More Characium pringsheimii from FY11-01-2016

I was able to get very good detail on what I am thinking is Characium pringsheimii.

I also found a couple of cells that had apparently ruptured and captured a small video of parasites feeding on the expelled inards.

Ankistrodesmus acicularis for Percy Priest Lake

I now have significant growth in the Percy Priest  Lake algae sample sample taken at the dam.

I examined the specimen under microscope and found several species of algae in the sample, including Ankistrodesmus acicularis.

Here are some videos and images taken 11-22-2016
Immobile Ankistrodesmus

Taken 11-25-2016 - showing more Characium and possible Euglena - doesn't look like Ankistrodesmus

FY11012016 Front Yard Algae Culture Upscale

Today I went ahead and upscale the FY11012016 Front Yard Algae Culture from 100ml to 2 liter using my standard MiracleGro media to see how well it upscales.

11/21/2016

12-03-2016 - ready to harvest??

We have algae culture growth in petri dish!!!

I went on a 3 day job and came back and found that I had algae culture growth in my petri dish simply placed in my newly created bioreactor.

AGAIN, the algae from the front yard pond grew successfully on the miraclegro algae agar from the recipe in my previous post from Doane College Algae Lab YouTube videos.

Here is a great link to all of their pertinent video series. Awesome resource!!
http://doanealgae.wixsite.com/algaegrowingresource/videos

And now the images of my petri dishes!! MESSY MESSY.

Being overly excited to cultivate a single species I have created 3 50 ml glass culture bottles made from Cracker Barrel maple syrup bottles!! I took small 'single colony' scrapping from 3 seperate areas and added to the bottles.

Here is day one images -

I placed the corresponding petri dishes behind the corresponding culture. 11-20-2016

Here is an image from 5 days ago - 11-15-2016. (significant difference in growth!!

I have Characium pringsheimii - i think

After observing my algae from the front pond, I dont think I have Euglena. I may have observed one or two but the large majority of the sample is more characteristic of Characium pringsheimii - http://protist.i.hosei.ac.jp/pdb/images/Chlorophyta/Characium/pringsheimii_2.html

Genus: Unicellular; cell body ellipsoidal or spindle-shaped, attached on a substrate at one end; a single chloroplast plate-like in shape, with a pyrenoid (Guide book to the "Photomicrographs of the Freshwater Algae", 1998).
Their image -

Secondary resource - http://www.plingfactory.de/Science/Atlas/Kennkarten%20Algen/01_e-algae/Chlorophyta/e-source/Characium%20sp.1.html

Their images - 

My microscope video

OMG Euglena eat algae!!! OH HAYUL NAW!!!

http://www2.fcps.edu/islandcreekes/ecology/euglena.htm

KINGDOM	Protist
PHYLUM	Euglenophyta
CLASS	Euglenophyceae
ORDER	Euglenales
FAMILY	Euglenidae
GENUS	Euglena
SPECIES	Euglena gracilis

Euglena gracilis and other euglena are green because they eat green algae. They keep the algae inside their bodies and use it to make their own food. These green parts inside the Euglena's body are called chloroplasts.

SO I HAVE IDENTIFIED AN ALGAE EATER IN MY ALGAE!!!

OH HELL NAW!!!

Using Microscope Attempt to identify algae species

Attempt to identify algae species using - - http://www.landcareresearch.co.nz/resources/identification/algae/identification-guide/identify/guide

Here are reference images of my summer algae sample that shows no sign of dying after 5 months.






And some videos

Show the dominate algae species is definately spindle shaped like this -

Single cellular with non distorting walls and no visible flagella.

My observation to answers to the identification guide 

1. Algae are unicellular - not joined to neighbouring cells
2. Cell wall is smooth, cell outline round to ellipsoidal, and featureless
3. Cells containing membrane-bound organelles, such as chloroplasts and nuclei.
4. Cell with rigid cell wall, lacking shallow groove at equator, and motile, propelled by one or more waving hair-like structures (flagella) - EXCEPT I CAN NOT SEE FLAGELLA, my microscope may just not be good enough to resolve - but had to say this to get to spindle shape..
5. Unicellular chlorophyte with an elongated, usually spindle-shaped cell with two short flagella emerging from one end. There is a single chloroplast that may contain a visible eyespot. If flagella are not visible, the cells could easily be confused with the spindle-shaped cells of other genera.

Final results says -

Chlorogonium (Chlamydomonadaceae)

looks almost identical - 

But NO FLAGELLA and the shape isn't exactly like mine - 

nothing on youtube on Chlorogonium.

sample is closer to - Euglena (Euglenidae) - http://www.landcareresearch.co.nz/resources/identification/algae/identification-guide/identify/guide/unicellular/euglena

You tube videos on Euglena viridis - much higher resolution than I can get. AT THIS TIME.

I wish I could afford a microscope as good as these!! DAMN!!

This video was recorded 400x!! I have to goto 650x just to see them. Meaning these appear to be alot larger!!

These videos show this species changing shape from circular to spindle and are not algae but PROTISTS!! - After observing with the 25x eyepiece instead of the usb camera - I noticed that there is definitely a split end that I can seen vibrating - meaning it HAS FLAGELLA.

Wikipedia says - https://en.wikipedia.org/wiki/Euglenid this about Euglena.

So at this stage I am waiting to see if the two samples I have in petri dishes come to fruition ...........then isolate and culture...JUST ALGAE, NO PROTISTS ALOWED!!!!

My Simple Square Box PhotoBioReactor PBR Design using recycled soda bottles

In an earlier post I showed my first PBR design from 2X4x and a leaning vertical tube design. While this design worked well. It was a real pain to detach and reattach the hoses at the bottom of the tube to clean or empty their contents.



Then I designed a semi-circular PBR using an instructable and alot of guess work to build a semicircular shaped pbr  and while it made removing and installing the tubes a little bit easier it was still too cumbersome and gangly to work with. Setting the lighting and air pump timers was a bitch. Also I dont think the tubes were close enough to the grow light.

Here are a couple of images for reference.

THE main thing I was having trouble with was using the vertical tubes, each time.
The last straw with the tubes was when I noticed that despite the air stones at the bottom of the tubes I was getting SIGNIFICANT setting below the air stones at the bottom of the tubes.

 

Again not very practical. It was fun to build but just sucked.

I found that every time I took a sample it was in a 20 oz soda bottle and for some reason just adding a 100 ml of miracle gro algae media I created earlier the algae grew better than in the tubes.

So I decided to eliminate the tubes and just use recycled soda bottles and design a simple easy to change and clean and move things around in pbr. I know a run on sentence.

Here is the first square design made from nothing more than 2x4s - oh I had to buy the plexiglass but the 2x4s I alread had, from the previous leanto design.

This design was WAY easier to use and more practical. I decided to try putting the light underneath.
This didnt provide enough light so I redesigned it to be enclosed ( slightly more aesthetically pleasing - and lighting from the side of the bottles for better light penetration.

Here is what I came up with - definitely not my final design - designing is the fun part!!

All from a 4'X4' sheet of osb, I had it laying around.



screwed together with 2x2s


Placed the air pump and timers on top for easy access.






Two 4 gang controls in back.
Video of prettier design in operation - I used mirror tiles to try reflect light back up towards samples.

So the dark green sample, Second from the left is the sample I have been keeping going every since summer. I am still yet to get anything to grow inside my incubator so I have also placed 2 more samples of my summer sample to the far left.

TIME TO USE MY MICROSCOPE TO IDENTIFY WHICH ALGAE SPECIES ARE IN THE SAMPLE, AND WHICH IS THE DOMINANT.  BUT I REALLY WANT MY COLONIES TO GROW SO I CAN ISOLATE ...........