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Polyculture Revisited

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  • Ron Brown
    Hello, All,        Yesterday, I took a gallon of warm water, three gallons of algae culture, and the 8 ounces of nitrifying bacteria, and put them in a
    Message 1 of 24 , Dec 28, 2012
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      Hello, All,
       
           Yesterday, I took a gallon of warm water, three gallons of algae culture,
      and the 8 ounces of nitrifying bacteria, and put them in a 5-gallon carboy.  I
      set up an air pump with a hose and bubbler-stone and plugged it in.  I started feeding
      it by adding one scoop of brown sugar (about a tablespoonful.).  I also added 8
      ounces of urine, so it would have some nitrogen to work on.  Today, I continued
      with this program.  I have been using some 5-in-1 aquarium test strips to make
      the daily tests.  The tests include:  General Hardness, Carbonate Hardness, pH,
      Nitrite, and Nitrate.  So far, the only change has been a rise in pH from 6.0 to 7.5. 
      The  culture looks good.
       
           As it turns out, nitrifying bacteria are not the same thing as nitrogen-fixing
      bacteria.  Nitrifying bacteria transform ammonia into nitrites, and turn nitrites
      into nitrates.  This is good for aquaria, but not so good as a bio-fertilizer.  What
      I really needed was Azotobnacter., which converts nitrogen from the air into forms
      that are usable by living organisms.  When used as a bio-fertilizer, Azotobacter
      increases yields by at least 6 %, but perhaps by as much as 60%.  They are aerobic
      and heterotrophic, so my current incubation strategy ought to work.
       
           I heard from the supplier in China, and the shipping charges for one kilo of
      Azotobacter is $98.00.  I e-mailed them with a big "Never mind."  (I can get a
      slant of Azotobacter from Ward's Scientific for ten dollars and change.)  So, my Quick
      Start is a _false start_.  It happens.  The ultimate solution will involve Azotobacter. 
      But, at least, I have gained some worthwhile experience in working with the Quick Start.
        
           So, the adventure continues.
       

      ---rsb
      Ron Brown
      28DEC2012
    • Ron Brown
      Hello, All,        I tried to order a slant of Azotobacter from Ward s Natural Science, but they have restricted sales of Azotobacter to schools and
      Message 2 of 24 , Dec 30, 2012
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      Hello, All,
       
           I tried to order a slant of Azotobacter from Ward's Natural Science, but they have restricted sales of Azotobacter to schools and research organizations.  I tried to call them on the phone, but they were closed for the weekend.  I went to Home Depot, and found their biofertilizer section, and bought some Kellog's biofertilizer.  It had about 15 different kinds of microbes listed, but I didn't recognoize any of them.  When I got home, I looked them up, and they all turn out to be fungi.  This was not good news...  I don't want fungi. 
       
           So, I did a little Internet Research, and found a site that said:  "You can buy  Azotobacter from any hydroponic store."  So, I went to Steve's Hydroponics to check.  Much to my surprise, they were open, and were glad to sell me 2 ounces of nitrogen fixing microbes for $15.99.  The microorganism is Azospirillium brasilense, which is not Azotobacter, but should be cloose enough.  The name of the product is Azos.  The attached "Azos.jpg" shows what the package looks like.
       
           I brought a gallon of the Quick Start culture into the house, and warmed it to about 70 degrees Fahrenheit, then added all the package of Azos to it.   I have decided to keep the Quick Start bacteria ( probably Nitrosococcus and Nitrospira.)  My nitrogen fixing bactgeria will convert elemental nitrogen into ammonia.  The nitrifying bacteria will cpnvert the ammonia into nitrates.  This seems like a good thing, because ammonia and nitrates can be poisonous, while nitrates are much less so.
       
       
      ---rsb
      Ron Brown
      30DEC2012
    • Ron Brown
      Hello All, I was able to grow Euglena, Chlorella and Spirillium together successfully. The symbiosis seems to make the combination unbeatable, but I have been
      Message 3 of 24 , Aug 15, 2013
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        Hello All,

        I was able to grow Euglena, Chlorella and Spirillium together successfully.
        The symbiosis seems to make the combination unbeatable, but I have been
        looking for something to do with the 2 1/2 acres that I have on the Salton Sea.
        The waters there are so nutrient-rich that it is causing ecological problems. In
        effect, the location is offering me free fertilizer for the algae, but the previous
        polyculture probably wouldn't be too comfortable in the salt water. So, I
        started looking for candidates for a new polyculture. So far, what I have come
        up with is this: Use Pleurochrysis carterae with Arthrospira platensis, because
        they both like pH of 11, and are both salt-tolerant. I would like to find a third
        'partner' that is aerobic (to get rid of the oxygen that ccmp647 generates), and
        preferably nitrogen-fixing. Apparently, Spirulina is not able to fix nitrogen from
        the air. A combination of elevated pH and salt-tolerance with nitrogen-fixing
        seems like it would be a bulletproof combination, especially with free fertilizer..


        ---rsb
        Ron Brown
        15AUG2013
      • dsieg58
        Ron, Is the end result to be fuel, or food, or both? Since Spirulina and Euglena have very little oil, it prompts me to ask. I hope this helps, David Sieg
        Message 4 of 24 , Aug 15, 2013
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          Ron,

           

          Is the end result to be fuel, or food, or both? Since Spirulina and Euglena have very little oil, it prompts me to ask.

           

          I hope this helps,

           

          David Sieg

           

          From: oil_from_algae@yahoogroups.com [mailto:oil_from_algae@yahoogroups.com] On Behalf Of Ron Brown
          Sent: Thursday, August 15, 2013 5:49 PM
          To: oil_from_algae@yahoogroups.com
          Subject: [oil_from_algae] Polyculture Revisited

           

           

          Hello All,

          I was able to grow Euglena, Chlorella and Spirillium together successfully.
          The symbiosis seems to make the combination unbeatable, but I have been
          looking for something to do with the 2 1/2 acres that I have on the Salton Sea.
          The waters there are so nutrient-rich that it is causing ecological problems. In
          effect, the location is offering me free fertilizer for the algae, but the previous
          polyculture probably wouldn't be too comfortable in the salt water. So, I
          started looking for candidates for a new polyculture. So far, what I have come
          up with is this: Use Pleurochrysis carterae with Arthrospira platensis, because
          they both like pH of 11, and are both salt-tolerant. I would like to find a third
          'partner' that is aerobic (to get rid of the oxygen that ccmp647 generates), and
          preferably nitrogen-fixing. Apparently, Spirulina is not able to fix nitrogen from
          the air. A combination of elevated pH and salt-tolerance with nitrogen-fixing
          seems like it would be a bulletproof combination, especially with free fertilizer..

          ---rsb
          Ron Brown
          15AUG2013

        • Ron Brown
          Hello David, ... This one would be for fuel, not food. I am kind of curious how close we can get to profitability in the real world for biofuel. The
          Message 5 of 24 , Aug 15, 2013
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            Hello David,

            >> Is the end result to be fuel, or food, or both?

            This one would be for fuel, not food. I am kind of curious how
            close we can get to profitability in the real world for biofuel. The
            ferrocement ponds can reduce capital expenditure a bit, nut free
            nutrients are even better than manure. Initial tests would be
            done in my small pond. Besides, CCMP647's use in human nutrition
            is questionable, although it DOES produce quite a bit of vitamin B-12.

            Since Spirulina and Euglena have very little oil, it prompts me to ask.

            This is less about oil yields and more about contamination prevention.
            Hardly _anything_ else thrives at a pH of 11, and both Pleurochrysis and
            Spirulina love it there. Pleurochysis is no slouch when it comes to oil yield: 30%-50% of dry weight. This is a step up from Chlorella, which only has about 30%. The effect of growing Pleurochrysis together with a filamentous alga could possibly facilitate harvesting, as Harshith Serigeri suggested. It seems like part of the puzzle is missing: CCMP647 generates oxygen
            during the day, and this depresses its growth. I think I need something that can deplete this oxygen. I have been looking at alkalipilic bacteria, but there is still no joy.



            ---rsb
            Ron Brown
            15AUG2013  








            .
          • dsieg58
            Interesting. You re right, PC is a great strain. If anyone can figure out the polyculture, it is you. I don t know if this will help or not:
            Message 6 of 24 , Aug 15, 2013
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              Interesting. You’re right, PC is a great strain. If anyone can figure out the polyculture, it is you.

               

              I don’t know if this will help or not: http://en.wikipedia.org/wiki/Oxygen_absorber

              In most formulations the active ingredient is powdered iron or iron salts

               

              Common oxidizing agents: http://en.wikipedia.org/wiki/Oxidizing_agent#Common_oxidising_agents

              ·  Hydrogen peroxide (H2O2) and other inorganic peroxides

              ·  Nitric acid (HNO3) and nitrate compounds

              ·  Permanganate compounds such as potassium permanganate

               

               

              David Sieg

               

               

              From: oil_from_algae@yahoogroups.com [mailto:oil_from_algae@yahoogroups.com] On Behalf Of Ron Brown
              Sent: Thursday, August 15, 2013 11:08 PM
              To: oil_from_algae@yahoogroups.com
              Subject: RE: [oil_from_algae] Polyculture Revisited

               

               

              Hello David,

              >> Is the end result to be fuel, or food, or both?

              This one would be for fuel, not food. I am kind of curious how
              close we can get to profitability in the real world for biofuel. The
              ferrocement ponds can reduce capital expenditure a bit, nut free
              nutrients are even better than manure. Initial tests would be
              done in my small pond. Besides, CCMP647's use in human nutrition
              is questionable, although it DOES produce quite a bit of vitamin B-12.

              Since Spirulina and Euglena have very little oil, it prompts me to ask.

              This is less about oil yields and more about contamination prevention.
              Hardly _anything_ else thrives at a pH of 11, and both Pleurochrysis and
              Spirulina love it there. Pleurochysis is no slouch when it comes to oil yield: 30%-50% of dry weight. This is a step up from Chlorella, which only has about 30%. The effect of growing Pleurochrysis together with a filamentous alga could possibly facilitate harvesting, as Harshith Serigeri suggested. It seems like part of the puzzle is missing: CCMP647 generates oxygen
              during the day, and this depresses its growth. I think I need something that can deplete this oxygen. I have been looking at alkalipilic bacteria, but there is still no joy.


              ---rsb
              Ron Brown
              15AUG2013  








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            • Ron Brown
              On Thu, 8/15/13, dsieg58@gmail.com wrote: Subject: RE: [oil_from_algae] Polyculture Revisited David, ... Thank you for the kind words. I
              Message 7 of 24 , Aug 16, 2013
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                On Thu, 8/15/13, dsieg58@... <dsieg58@...> wrote:

                Subject: RE: [oil_from_algae] Polyculture Revisited

                David,
                >>Interesting. You’re right, PC is a great strain. If anyone can
                >>figure out the polyculture, it is you.  

                Thank you for the kind words.

                I think I've got it! Botryococcus braunii could serve as the
                oxygen scavenger. It likes high pH, and is salt-tolerant. The
                fact that its oil. is not the kind we want is probably not a
                problem. Its slow growth shouldn't be a problem, either.
                Since this will be in a raceway, shear stress won't be an
                issue. BB can also do heterotrophy when it puts its mind to
                it. The only thing lacking is some kind of nitrogen-fixer,
                but that may not even be necessary.


                ---rsb
                Ron Brown
                16AUG2013




















                .
              • Bobby Yates Emory
                Ron, Pc is a great choice (I m sure no one is surprised I said that.) Since you are using a raceway, if you have a paddlewheel (or substitute), it should
                Message 8 of 24 , Aug 16, 2013
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                  Ron,

                  Pc is a great choice (I'm sure no one is surprised I said that.)

                  Since you are using a raceway, if you have a paddlewheel (or substitute), it should "splash" the O2 into the atmosphere.

                  Bobby


                  On Fri, Aug 16, 2013 at 5:09 PM, Ron Brown <potrzebie44@...> wrote:

                  On Thu, 8/15/13, dsieg58@... <dsieg58@...> wrote:

                   Subject: RE: [oil_from_algae] Polyculture Revisited

                       David,
                  >>Interesting.  You’re right, PC is a great strain. If anyone can
                   >>figure out the polyculture, it is you.  

                       Thank you for the kind words.

                       I think I've got it!  Botryococcus braunii could serve as the
                  oxygen scavenger.  It likes high pH, and is salt-tolerant.  The
                  fact that its oil. is not the kind we want is probably not a
                  problem.  Its slow growth shouldn't be a problem, either.
                  Since this will be in a raceway, shear stress won't be an
                  issue.  BB can also do heterotrophy when it puts its mind to
                  it.  The only thing lacking is some kind of nitrogen-fixer,
                  but that may not even be necessary.


                  ---rsb
                  Ron Brown
                  16AUG2013




















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                  --
                  Toward freedom,

                  Bobby Yates Emory
                • Ron Brown
                  On Fri, 8/16/13, Bobby Yates Emory wrote: Bobby, ... Yes, it is. Strange that it should require free food to bring it about. ... Well,
                  Message 9 of 24 , Aug 16, 2013
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                    On Fri, 8/16/13, Bobby Yates Emory <liberty1@...> wrote:


                    Bobby,
                    >>Pc is a great choice (I'm sure no one is surprised I said that.)

                    Yes, it is. Strange that it should require free food to bring it
                    about.

                    >>Since you are using a raceway, if you have a paddlewheel (or
                    >>substitute), it should "splash" the O2 into the atmosphere.

                    Well, some of theO2 may leave, but More O2 and CO2 will enter,
                    so I don't thin k the paddlewheel is a complete solution to the problem.
                    If memory serves, Navid's research involved a paddlewheel, but he
                    found fluctuaating O2 levewls in the culture.


                    ---rsb
                    Ron Brown
                    16AUG2013






























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                  • Ron Brown
                    Hello All, In my last post on this thread, I said: The only thing lacking is some kind of nitrogen-fixer, Well, I think I have found a suitable candidate:
                    Message 10 of 24 , Aug 21, 2013
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                      Hello All,

                      In my last post on this thread, I said: " The only thing lacking is some
                      kind of nitrogen-fixer,"

                      Well, I think I have found a suitable candidate: Synechococcus. This
                      is an aerobic marine alkaliphilic nitrogen-fixing cyanobacterium that can
                      tolerate pH of 11.


                      ---rsb
                      Ron Brown
                      21AUG2013
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