sgtrock101: If this experiment works I will repeat a third time. That should be confirmation that the algae/bacteria (from septic water) consumed the nutrients in the water, including visible particles of waste matter
Hi sgtrock,
I dont think the algae have anything to do with what you are seeing. Particles will disassociate into solution all by themselves. Bubbling air thru it will quicken that process up.
To test this hypothesis... could you not add algae to your nutrient solution in one flask and add algae to another?
flectere si nequeo superos, Achaeronta movebo! -Virgil
Yea, that works. I have a second sample of the mongrul just with soil water. The amber hue of the water lightens up as the algae grow in numbers. The algae does not consume solid particles, as I understand algae. So, I believe that the particles are being broken down by bacteria from the waste water and then the algae consumes the waste. The clarity of the water and the increased mass of algae in the waste water sample leads me to believe the particles just do not go into solution and stay there. The soil water nutrients could not support the increased mass of the newer algae. In soil water only the mongrul algae grows slowly. With the waste water the growth is much faster. The clarity is what gets me. It does not take long for the water to clear, aprox 1 week. I was expecting to see some of the particles form a sediment. Well, I used unfiltered waste water this time. After one day the water appears to be clearing, certainly not as cloudy as when waste water was first added yesterday morning, with no sediment forming. I'll be able to know in about six to nine days. If this does clear and the algae mass is noticable increased, a third confirming experiment should tie it up.
I reread your comment and have a better understanding of what you suggest. Right now that is not possible. I have a limited amount of aquarium pumps (I'll buy more latter this week). As I understand it, you suggest I use the air pumps with a solution of soil water and waste water only, and the same with algae added. This to determine if the particles dissolve into a clear solution, air pump aided, without the influence of algae. I'll try that within a week to 10 days when I have more aquarium pump capacity. It's a good idea and will either back up what I think is happening or turn my idea into ---WASTE WATER!!!.
Hey Sg. Spending all your $ on pumps? lol.
Where you live? Do you have a microscope?
Im quite sure that the algae/bacteria have almost nothing to do with the water getting clear and quite sure that it has everything to do with chemistry into solution.
Here is why this is important and why I think biowaste streams like from a dairy farm is a very poor choice for feedstocks. Because those organic microscopic and submicroscopic molecules of sugar and proteins and lipids and biomass pieces and parts from the wastestream turn into very small solution in a very short time. And because algae dont need them but bacteria and other nasties do, this is why a monoculture can likely never be achieved. The solution is to remove all organics from the instream water source of the system and you can keep virtually all other organism except other autotrophs at baye. We can work on autotrophs competition later...
Additionally, the system that takes biowaste and digests it in water is quite likely creating methane at a much higher rate than a normal system. CH4 is by far worse for the environment than CO2 by 40x. Thus a 2.5% shift in the C output to CH4 over CO2 is clearly more pollution.
IMO, manure and human wastes and other particularily bad organic materials that humans have made need to be gasified/pyroliside/plasmide/chemicalized and any other kinda ide you can show me you can do to reduce virtually 100% of the C to CO2 or something else sterile. No more e-coli, no more feeding raw blood to chickens, no more letting AD systems for MWS then landspread the rest. No more paperpulp sludge. It comes out of the cows butt, it goes to a conveyor belt and is turned into e' or other before the cow lets go of another load. You can then have the ash which will be most of the input nut's you will need because ash doesnt burn. Remember, autotrophs hate fixed carbon and love ash. Sprinkle in bit of makeup ash and some N products and your rich because you are getting a two-fer. They are;
You are feeding your cow on either 10-20x less acrage/present output or 20x more cows/existing infrastructure.
You are feeding the grid e', cleaning up CH4 and disease pollution and creating the feedstock the process needs to start all over.
Actually that is more like a ten-fer because you have a hard time growing autotrophs in sewage so one of the 'fer's' would be that you could ACTUALLY have a shot at mass producing something someone wants, food.
One more 'fer'. I would be afraid of the organisms that live on the backside of an organic sewage digester.
Froggy, I have a microscope, must be more than 30 years old, x200. It is not adequate for what I am doing. I want to see images like some of the internet sites have where the cells are large, translucent and the potential lipids visible. I also need to have the scope able to take digital pictures of what I am seeing (or allowing me to put a camera eyepiece up to the scope eyepiece and take a picture. Some on this forum have been able to do that). As a side bar I want to use one of the methods for aproximating lipids by red nile die and florescent light (and photograph that). Quite a set of needs for a man on a tight budget. I'll get there, it just takes time and patience.
I live in the New Orleans, La area, a suburb. There are lots of medical types in the city. But, they have an attitude problem (arrogance) I have difficulty dealing with. So, I stay away from them.
I examined the samples again today. The 1st 1750ml Bb sample continues to grow, with the white mist pulsing intense every couple of days. More cell growth occures and become visible. The total estimated growth is aprox 50% more tha last close observation. This sample continues to catch up with the growth of the second 1750ml sample of Bb.
The second sample of Bb is expanding rapidly. There is much more cell growth clinging to the sides of the plasdtic jar and air tube. I have repeatedly removed the cell growth from these attachments over the days, but the cells return and reattach. Some even travel up the atmosphere providing tube. They are a little bit of trouble to remove and restrict the airflow. A curiosity. The majority of the cells combined into a round mass aproximately 1 1/2 to 2 " diameter ball. This ball flows with the current , is picked up by the atmosphere outlet point, goes to the top of the fluid level, floats along then sinks into the current and repeats the travel. I had observed this before a few weeks ago. If it follows previous bwehavoir, it will disperse into clumps, the heavy misting will return in pulses, and much more cells will be visible than before, and the cycle startes again. It seems to me that the cells communicate with each other when gathering. They may do so prior to combining into a ball. This reminds me of insect colonies, like bees. When undisturbed, like bees, things hum along, groups form and do something, then disperse to reform into other groups later. This is the 'happiness' I characterized on an earlier post.
The mongrul samples are doing good. The smaller sample, without the waste water, is not growing much. However, it has returned to a vivid bright blue green color. Previously, when it was growing, it was a much duller color. The medium hue has almost cleared to no amber color. I suspect I will have to recharge the sample soon with more soil water. The second mongrul sample, the one with unfiltered waste water, is clearing the sample slowly. It is also growing. I will know for sure when the water is at its best clarity.
I will buy the aquarium pumps I need today for additional experiments. I want to perform the experiment froggy suggested regarding waste water.
As always, comments that are relevant are welcome at anytime.
The Bb sample that had rolled itself into a ball broke apart today, as it has done before. If it does as before the misting will increase and more cells will appear. I still think the mass of Bb has a communication mechanism, a system that causes it (the many cells and clumps of cells) to gather, do whatever, and then disperse. Has an yone observed this behavoir before.? Does anyone know how long the cells live? Right now there are no dead cells visible. That means the vast majority, more that 99% are more than 3 weeks old.
The mongrul sample with unfiltered waste water is becoming more clear and there is a dusting of waste particles settling out. I have shaken the sample ad caused toe sediment to disperse. The air agitation is not disrupting the settling. In a few more days I hope to see the sediment disappear as the previous waste water (filtered) had done. The algae seems to be multiplying.
Relevent comments and questions are invited.
My first time in this forum. Lot of exciting ventures and adventures of folks all over.
I am in PA. I am growing nannochloropsis sp. and Botrycoccus Braunii.
Vigorous and constant aeration produces geen but blue tinged dense culture of BB with a few clumps. Like mentioned elsewhere co2 can decrease ph and affect BB growth which likes alkaline environment. Also inhibition by its own metabolic products could be another reason for discoloration. Transfer to fresh solution often will produce consistent results.
I am able to supply living samples of these algae for a reasonable cost+shipping and it will be much denser than UTEX.
I am now looking for a low cost lab scale ultrasonic homogenizer...any leads will be welcome.
Jay,
What were you thinking of for BB costs? I am in N.C.
Hi Liberty1
I think I can ship 100 ml of concentrated BB to North Carolina for $50/- . This should be enough to innoculate b/w
1litre to 5 litres of culture solution (F/2 guillards)
compared to UTEX barely can see for $70/-
Kindly let me know if you think this is a reasonable price....This is my first time and I am not trying to make a lot of profit
but be able to provide to all seekers at reasonable costs...
Thanks
Glad to hear someone else is growing Bb. What strain is your Bb? When you refer to alkaline, what are you using to get this alkaline and what ph looks good? When refering to changing water frequently, what is your interval? What medium are you using? I use a soil water formula that Marc had posted on the boards.
I've been experimenting with a mongrul algae and a canine septic waste system I built. I'm hopping to use the septic waste water as a source of nutrients.
My samples are going on. The UTEX sample has multiplied greatly over the last six weeks. The 15 ml sample just barely filled the rounded bottom of the sample tube. Now I estimate I have 30 to 40 ml, more than enough to fill two 15 ml tubes completely with a wet mass. The growth and color change to a dark olive brown is not mentioned by anyone in research reports. But, the samples keep growing. So, I'm a little confused but, hey the samples are growing. Is it the nutrient medium?
Hi sgtrock101
I am a chemical engineer , I have asked our biology guy which strain it is and I will let you know when I find out.
We keep ph at 8 and do not bubble co2, only air. we use f/2 guillards medium acquired from aquaticco.com.
I am afraid brown is not a good color for photosynthesis but I will let the biology people fight over that one.
Look under the microscope to see if there are any foreign bodies or cell damage..
Bb is pretty slow growing (compared to nannochloropsis), it takes a 5-6 days for the medium to reach the growth plateau.
Do not quote me on this because there are way too many variables, use a dip disk to measure the algae density to find out your own plateaus.
This is a good time to change the culture.
I tried aquaticco.com but got an error message. Do you have a different version?
Sorry Bobby, a typo, here is the exact link to get F/2 guillards medium
http://www.aquaticeco.com/subcategories/1364/F-2-Algae-Food-by-ProLine/algae/1
Good luck
Thanks for the specific URL. I was unable to find it by searching around.
But I did find something someone asked for - a dry algae food:
http://www.aquaticeco.com/subcategories/1367/Microalgae-Grow-Mass-Packs
(Note: Most of it is dry, but there is some liquid included.)
This looks like a great deal. Has anyone checked it out?
Watch your shipping costs when ordering from Aquatic - I was shocked.
A new Bb experiment has been added. I bought some bone meal at the local hardware store, nursery section at Home Depot. I added just two gramuals to one of the 1750 ml samples. In just 48 hours - WHAT A DIFFERENCE!! At first everything was just lazy looking the first 24 hrs. Then today the white misting has increased and many many more cells are visible. There are many new clumps of cells and they are, for want of a better phrase, buzzing around the medium. It almost looks as though they propel themselves, some moving with the current and others crosswise to it.
The new experiment is a 500ml sample with a small clump of Bb, aquarium pumped normal atmosphere filtered thru a A/C filter, and four granuals of the bone meal. I started this six hours ago. The clump looks much more lively. The clump has expanded to resemble a snow flake, or a ragged sail catching the wind. I think the cells are increasing the clumps surface area to gain greater exposure to the bone meal (nitrogen and phosphates) now in solution. I am really surprised at how quickly this is occuring. To me this is very exciting. I started with such a small quantity of bone meal hoping not to over "feed" the algae.
So, the first 1750ml Bb sample has had two bone meal granuals added to it 48 hrs ago and already much activity is noted. This first sample was the one that had the lesser amount of cells from a prior experiment, it was slowly but visibly catching up to the 2nd Bb sample that is without bone meal granuals.
Up to this point I conclude that container shape, volume and ariation by aquarium pumps all have a visible impact on the Bb. Generally speaking a large container ( aproximately 2 leters, square shaped , 4"x4"x6") with aproximately 1500 ml, or more medium, that is areated with an aquarium pump ((Manf Aqua Culture mdl MK1501 - made in China, where else?- 5-15 gal max capacity 1200cc/min thru a single outlet aprox 1/8" dia) continuopusly operating creates a gentle yet forceful current that cause all the cells to be exposed to the medium. To me, they (the cells) seem to enjoy this. They wll mist, clump into smaller cell groups then roll into a much larger ball, then disengage into small clumps again and again. As nutrient is the soil water medium are consumed the amber hue of the medium becomes much lighter. As the medium nutrients are consumed the cells will act more lazy as the nutrient become less concentrated. The clumps look much more tight, as though the cells groups were protecting themselves from a predator.
The addition of the bone meal is what is so revealing. To see the cell clumps 'unfurl' to expose themselves to the medium is very dramatic. As yet I have seen no visible dead cells and the cell color remains a dark green/brown. I am still wondering how long does a Bb cell live? If it is long lived than a dairy farm system of extraction of cell lipids makes sense.
The mongrul samples are doing as I had predicted. The waste water has been consumed, or gone into solution, is no longer visible, there are much more algae cells present and there is only a film of sediment in the upper regions of the flask out of the body of the medium. The control sample is alive but not growing. It has rolled into a couple of tight balls and just floats in the current.
I will begin a 3rd experiment with the waste water and medium solution, areated, without algae, as suggested by froggy. I will observe what happens to the waste water, see if it might desolve into solution.
The dramatic change in the Bb sample with the added bone meal is the big event for me. It remains to be seen how much larger the sample size will become. Hopefully the cells will visibly multiply to a point where I will need a bigger container to hold them in.
As always, relevent comments are welcome and encouraged.
Long time since last post
There are three Bb colonies. There had been four. The forth died, I believe. The dead colony was over feed bonemeal and turned ocher brown. Everything became too heavy to flow in the current. All cells drifted to bottom of 1750 ml jug. No amount of shaking, stiring or turbulence created by aquarium pumps keeps any cell groups afloat. It had just ceased to do anything. The water clearede up and is easily viewed thru. I have ordered a new more powerful 400x microscope and will examine the cell groups to see what dead looks like. I have not disposed of the sample until I can confirm death. I will then filter out sample and store it in freezer. When I have the tools to use nile red stains I will crush it up and get an approximate oil density by flourescence.
The other three Bb colonies and two mongrul samples are doing well. The two 1750ml and 500 ml colonies have all stabalized with slight clouding, They all see to be in a state of equalibrium. I have bought f2 growth medium from aquatico.com and am slowly introducing it into medium. The only real change since introduction is the color. All three have grown dark green again. Previously they had been a dark brown green. The clumps within each colony appeard more dense and there are many clumps floating in the current. When the bone meal granuals were added the clumps appeared to unfurl like a sail but now have tightened up into denser masses. Not quite round but not totally unfurled either.
The mongrul sampls have shown the greateast increase. The canine waste water just caused an explosion in its two samples. So dense that it is a dense cloud when stirred and not able to see thru. When not in the current stream it has formed a dense carpet on the bottom of the jar. Very fast growth. Now, it too, is in a state of equilibrium, the same as the smaller control sample. Color is a very dark green. The cells are very heavy and quickly fall out of the current after being stirred. I wonder if there is any oil in the cells?
I am concluding that, for the mongrul sample, the canine waste water is very effective for growth. After I clear out the dead colony I will begin a sample colony of Bb in that container, 1750ml with f2 medium and see well and quickly it grows. Co incident I will buy another jar and use waste water with Bb. It should show a good comparison of how the different mediums affect growth.
I began the Bb in wastewater experiment today. I placed aprox 20ml of canine wastewater into a jug of 1750ml with a small Bb sample. I am hoping for the same rapid growth I saw with the mongrul algae sample experiment.
Everything seems in limbo right no. Noe visible growth but no die offs either. The fe mediums have added a lot of green color to the Bb and mongrul samples. It may be the temperature that is holding things back. The locale has experienced 95 deg F + days for several weeks.
As always, related comments are encouraged.
Hi! my name is Ailin I am Biothecnologist and I've been working with B. Braunii for more than one year. This observations that you says I noticed to. About its culture or ways to optimizaded the medium its depended what you need or for what you want to use Braunii. But if you have been watching a lot of diferences in its growth in the same culture you have problems, you must control the variables as much as posible.And then you can observed that they growth in conditions that you can reproduced easily. One part of my work I published in a congress this year, but maybe in a couple of months I am going to public all the results that can help you.
If you need something especialy just write.
Ailin Beznec