phytoplankton culture help
 

ok i got a setup from a member on here, its the whole setup, 3 gallon set up my ? is do i fill it up all the way with saltwater leaving enough to put 500ml of phytoplankton in and the f2 fert, and when i split do i leave half in and put new salt water in and the fert to keep the culture going, and do i have to put the split cultures in the fridge, sorry guys a lot of ?? i just don't wanna mess things up
thanks

I'm assuming that you are talking of the mamaroo phytoplankton kit.

and the air line and light.

I wouldn't fill it to the brim.
Leave a few inches for the bubbles to pop against other wise it will leak unless you fully tape the lid on to it. If you do that, you will end up with a sludge that forms behind the tape and has to be cleaned everytime you open it up.

yeah pretty much
read up on the amount of f2 required. It's not a lot.
http://www.melevsreef.com/phytoplankton.html

I always have, not too sure if you must.

is it going to be ok if i have it in my fish room or does it need complete darkness, cause my sump light is on 18hrs a day

yes it is the mamaroo kit

To maximize growth rate, light the phytoplankton 24/7. It does not need a period with the lights off. The phytoplankton gets its "dark" time as the cell circulates in and out of the inner parts of the culture. I'm not sure what species you have, but typically 100 umol (PAR) is a conservative light intensity for microalgae.

Also, this website details the F/2 medium and gives some information about general algae culture.

https://ncma.bigelow.org/node/79

F/2 can be purchased in bulk from aquatic ecosystems. The 1 gallon jugs of part A/B would last most hobbyists a very long time. To make it easy measuring-wise, you can make pre-mixed "stock-solutions" and keep them in your fridge. This would allow you to add (for example) 1 mL of stock solution per L of culture to achieve the appropriate concentration.

http://www.aquaticeco.com/subcategor...F-2-Algae-Feed

so i have to keep my f2 in the fridge , and why does everybody say 16hrs light and 8 h darkness iam confuzed

I always get best results 16 hours on, 8 hours off.

myka do you keep yours in the fridge too

The f/2? Yes. I haven't had any luck with f/2 though, I think I got a bad batch. I've been using Miracle Grow and Kent Essential Elements, but I worry that f/2 might make the phyto more nutritious, so I'm going to order new f/2 and try it...eventually.

I don't put any phyto in the fridge.

Yes you fill the reactor with fresh saltwater mix
Then you add the 500ml included and fertilizer
:)
When is time to harvest you can split the half
The rest you can keep it in fridge or sell it ;)

You are likely using too intense of light then. From a physiological standpoint there is no reason to give microalgae 8 hours of darkness. The dark reactions occur on the order of milliseconds and the light reactions occur pretty much instantaneously. The cells get their "dark" period on the inside of the reactor, when they are not illuminated. If you give the algae 8 hours of darkness, they are likely switching to respiration, which will lower growth rates. Try reducing light intensity to 30 umol on a 24/7 schedule, and then slowly ramping up the light intensity over time.

As a plant to grow properly you need to provided the right habitat, which depends of the species they need usually a light intensity of 2500-5000 lux (250-500 footcandles) Conversion of foot candles to lux or lumens is: one ft. c = 10.8 lumens or lux, for nannochloropsis oculta (the mos common phyto to culture) you need 2,500-8,000 lux...You would get that intensity in a led warm white light on a 16 hours on that will allow the micro algae to breething co2 and you need 8 hours in full darkness for breething oxygene...like this system you will have a balance with your PH level inside the reactor chamber. Light shading by algae cells would become limiting as density increase, at high densities incoming light is shade from all but the cells currently at the outer surface of the reactor, that is mean is time to harvest the culture.

Microalgae do not require a prolonged period of darkness to "breath oxygen". Microalgae require ATP to fix CO2. They produce ATP only when illuminated, and therefore will not fix CO2 in darkness. As soon as CO2 fixation ceases, growth will also cease. I'm not saying you can't grow microalgae if you give them a dark period, I'm saying growth is maximized if they are illuminated 24/7.

Here are a few references that explain the reasoning, which is firmly based on the well established physiology of plant cells and photosynthesis.

"..long dark periods (i.e., of the order of magnitude of several hours) generally result in biomass loss, as well as decline in growth rates, because microalgae switch to respiration processes; several authors have indeed suggested that a suitable dark period should be of the order of miliseconds (Kommareddy and Anderson 2004)—which would be more appropriately seen as an intermittent cycle"

From - Carvalho et al. 2011. Light requirements in microalgae photobioreactors: an overview of biophotonic aspects. Applied Microbiology and Biotechnology, 89, 1275 - 1288.

"This internal shading (clearly visible in that light does not pass through the culture’s optical path, being essentially fully absorbed in the outer surfaces), results in cells receiving light intermit- tently, a phenomenon augmented by the fact that light energy attenuates exponentially in passing through the culture column. The higher the cell density, the shorter the depth light penetrates into the culture. Two light zones are thereby established in the culture: the outer illuminated volume, in which light is sufficient to support photosynthesis (i.e. the photic zone); and the dark volume, in which net photosynthetic productivity cannot take place, since light intensity is below the compensation point (Fig. 8.1). The higher the population density (and the longer the optical path), the more complex it becomes to address the basic requirements for efficient utilization of strong light, i.e. an even distribution of the available light to all cells in the culture, at an optimal dose per cell (to be elucidated somewhat later).

Clearly then, when mutual shading prevails, cells are not exposed to continuous illumination but rather to cycles of light and darkness (L–D cycle), which may take scores of milliseconds to a few seconds to complete, depending on the optical path and the extent of turbulence in the culture. The endless combinations of light intermittency expressed in L–D cycles to which the individual cells are exposed at a given instant, relate to two basic para- meters: first, the ratio between the light and the dark period in the cycle and second, the frequency of the cycle. As shall be elucidated, the higher the frequency of the L–D cycle, the more efficient strong light may be used for photosynthesis."

From Richmond (2005) - page 127/128. "Handbook of Microalgae Culture".

Jakegr,
Thanks for posting references.
It's helpful to see that.

I am not say that you can't grow microalgae if they are illuminated 24/7. I just said that it would be unhealthy, the phyto required the most nature habitat you can provided. Lets start with a natural law "everything that is over used is bad from a carrot to exced of exercise" I am analizing what you said and the books part that you used, so if phytoplankton grow good and normally being under lights 24/7 of the 12 months of the year how is possibly that phytoplankton have declined in the world's oceans over the past century in response to ocean warming. When the phytoplankton have abundant sunlight, carbon dioxide and disolved nutrients an undergo explosive growth forming blooms, algal blooms, during these blooms most of the phyto die and sink to the bottom, where descomposes. This process deplets the dissolved oxygen at the water bottom which demaged the culture in no time, in the ocean time this occurs during the springtime.
For confirm what I just wrote you can check on wikipedia: algal bloom; earthobservatory.nasa.gov/NaturalHazards/
There is many reason you don't want to have a wrong amount of light for your phyto the cells will get dense and they would don't feed certains organism like tisbes or rotifers.
In bade the hyphotesis you used, you also can have a look at the following paragraphs on the same book, I just will writte a part!
Appl Microbiol Biotechnol (2011) page 1279; Kimmareddy and Anderson 2004:- In view of the relatives magnitude of aforementioned time scales, the turnover time of the photo-synthetic unit (PSU) or photosyntetic reaction center, is given by the dark reaction time for practical purposes. The light-dark cycle period, which is determined by the travel time of cells between the dark and lit portions along PBR, should accordingly be made AS SHORT AS POSSIBLE, this usually means an optical path of 0.5 to 1.0 cm (to make it clear if the dark time was not necessary then they would mentioned)

Appl Microbiol Biotechnol (2011) page 1279 "Effects of excessively low and high light supply"
"Althought light is required for photosynthesis, too low or high levels thereof will entertain serious disaventantages!! ..."
To who is interesting this pages could be read at http://www.hcmv.vn.refer.org/moodle/...ic-aspects.pdf

You can have more information on Plakton Culture Manual by Frank H. Hoff & Terry W. Snell.

http://en.m.wikipedia.org/wiki/Phytoplankton
Also you can read it all, but if you just want to be more secure about the light process, look on the last paragraph of Aquaculture.

Just to make it clear, it is always good to try or do different things, for some reason what you suggest didn't work for me on one of the Golden Brown Species of phyto which one required a strong illumination. I am just try to pass on all the information I learned and help any one who is interesting, I have six species of phyto and other live foods and I learn to don't expouse your culture to something that may don't work or you are not 100% sure, but I still respect all the opinion of other people. 16 hours on and 8 hours off it works for me since I start the cultired.

thanks all for the input, iam learning as i go

My method works very well for me. I use less electricity and get the same results as 24 hour lighting. Phyto doesn't need a dark period, but it also doesn't need 24 hour lighting. Thanks for the advice though. :)

You are misinterpreting what that first reference is saying. It is saying that the light-dark cycle should be repeated all day by circulating the culture vigorously, not that a long dark period is helpful. I never did say the dark time is not necessary. I said it should be on very, very short and that they get ample dark periods in a dense culture by moving in and out of the "photic zone" of the culture.

I do not doubt you have been culturing phytoplankton for a long time with good results. So have I. You obviously have a solid practical knowledge about how to do it, but based on your explanations you do not understand the physiology. If I'm wrong, just answer this simple question:

As you said, from a physiological standpoint, why do you think microalgae need a prolonged dark period to "breath oxygen"?

That just doesn't make sense. Plant cells produce oxygen during photosynthesis by splitting water. Outside of the chloroplast, cellular respiration is occurring and uses up some of that oxygen (although there is a net production of O2 if light intensity is high enough). This is a major advantage of having a chloroplast - it provides oxygen to the rest of the cell. So why would they need a dark period to "breath oxygen" if they actually are able to obtain plenty of oxygen internally when illuminated?

We are not talking about multi-cellular plants here, which have many reasons to need a dark periods. Microalgae can be seen as the most basic photosynthetic unit and there is no physiological reason microalgae need a dark period. It is not "unhealthy" - I'm actually not sure what you mean by that, as it is a pretty vague statement. What process is being disrupted by illuminating with 24/7 light?

Again, I'm not saying you should switch methods as your method works for you. But I do feel if incorrect information is being spread, which is a problem in our hobby, that it should be corrected.

I do not think global warming has anything to do with our conversation.

Jakegr I am not taking my information basing just on one book, you keep saying illumination should be 24/7, I don't understand on which time you get the dark time then! You don't explained the intensity of the illumination either...I understand all the reading, I just highlighter the point that you didn't mentioned where clear said "Effects of excessively low and high light suppy" Algal bloom, as I said read those pages plus Plankton Culture Manual and the last part of Aquaculture on Phytoplankton from wikipedia were it said "Light must be provided for the growth of phytoplankton. The colour temperature of illumination should be approximately 6,500 K, but values from 4,000 K to upwards of 20,000 K have been used successfully. ¡The duration of light exposure should be approximately 16 hours daily; this is the most efficient artificial day length!!"
I am sorry I put breath oxygene instead of consuming oxygene, I just see that know, I am going to fix it "Algae produce oxygen during the daylight hours and then consume it at night. During the day algae use energy from the sun to drive a chemical reaction between carbon dioxide (CO2) and water (H2O) to produce sugar and oxygen (O2). The second respiration process in plants is called dark phase respiration. It occurs primarily during the night and, to a much lesser extent, during daylight hours. During the dark phase process the plant consumes sugar and oxygen to produce energy, carbon dioxide, and water. The critical point is that at night, aquatic plants shut down the oxygen- producing photosynthetic reaction and initiate a process where they consume oxygen. I feel 24/7 is unhealthy because we try to provided for our aquarium a live food that have to be proper culture to give the right nutrition. My goal is to describe the technical aspects of culturing phytoplankton, I am try to have a simple description that should be comprehensible to those with minimal experience in aquaculture, how you can see I am not just have a succes experience with one type of algae, but to 6 and I am not just try to make points but also to promote more species and the proper care. If you feel the information what I provided have been incorrect, then you are free to contact Marine Biologyst in Chile and Argentina to correct them, also you can contct Florida Aquar Farms, Inc to they stop sell the Plankton Culture Manual, you can also leave a disagreed feedback on wikipedia on phytoplakton Aquaculture information. Just analized how the phyto works in the ocean.

Donkey77 I year and a half ago in the city where I live, Edmonton, just 2 stores would provided phytoplankton one species "nannochloropsis" I have the pleasure to see mamero works he not just culture Nannochloropsis, he also culture other species, rotifers, brine shrimp and pods. He don't used the coca cola bottle system, he use a container and when you look at it you can see his deditation in the culture. This guy is a genius, beside any time I being on his home I saw how he take time to explained and listen to the people, I walked more then a time with extra free stuff and I do have wild corals that seems to love his phyto. The bad things in pages like this one is that every one can post copy texts and try to make points destroying the work of others. I will take any opinion or words from Mou as I used and see his live food culture system.

Jake I will like to know which specie or especies of phyto do you culture, I also will like to know if you have another type of life food culture. I will also like if you can post pictures of your system. I think actions talks more then words, I don't think any one in this Country have import and done the job that Mou did, when we try to help out comunity we don't need just talk we also action, I didn't see any store in the Country that provided many species of algaes as Mou that is why I keep saying he is a genius! He informed himself on how to proper care of the live stock and come up with something that not ones have in an entire country, now I don't think so he brought something and he wouldn't know how to take care of it.

OK, I will start at the beginning. Hopefully I do a better job describing it below.

1) The dark reactions are a misleading term that really is not used much anymore, although many scientists still say it out of habit. It leads people like you to believe these reactions can only occur in the dark, when that is not true. These reactions occur only while the light-dependent reactions are producing ATP, since ATP is required to fix carbon. They are more accurately called the "Calvin-Benson Cycle" or "Light independent reactions". There are examples of carbon fixation only occurring at night, but they are in multi-cellular plants often called "CAM Plants" (this is a mechanism is prevent water loss in hot terrestrial environments).

2) Inside a dense algae culture, there is a "photic zone" (light) and a "non-photic zone" (no light). Algae cultures are constantly being circulated, usually using an air pump. This transports microalgae cells between these two zones. Since the light-dependent reactions occur on a nearly instantaneous timescale, the rate limiting part of photosynthesis are the light-independent reactions/ Calvin cycle. All photosystems in a microalgae cell can become saturated instantaneously at high enough light intensities, meaning there are not enough electron carriers to process any more incident photons. These electron carriers are re-generated during the Calvin cycle. This is the physiological reason why the Calvin cycle limits photosynthetic productivity.

3) Therefore, if the goal is to maximize growth rates, ideally the algae culture will be circulated such that each cell spends milliseconds in the non-photic parts of the culture and nanoseconds in the photic zone. This ensures a constant supply of electron acceptors to process incoming photons. So, finally, to answer your question: the dark time occurs when the algae cells are in the non-photic parts of the culture. If you want to maximize productivity, this dark time should not be hours, it should be milliseconds. This is achieved by transporting the cells in and out of the photic zone really quickly via circulating the culture with air bubbles, NOT by turning the lights off for several hours.

4) If you do turn the lights off for several hours, you are only regenerating the electron carriers ONCE! Once the lights are off, those electron carriers are no longer needed for photosynthesis, and will not be useful until the lights are turned back on again. Once the lights are off, the algae cell will no longer be producing sugars from photosynthesis. Instead, it will consume them via cellular respiration in order to keep the cell alive. This is obviously not a productive time for the algae culture. It does not keep the culture "healthy".

You seem to be confusing photo period and light intensity. Light intensity is the number of photons hitting a unit area per unit time (PAR = umol photons per m^2 per s). Photo period is the length of time a culture is illuminated.

Excessive light intensity can cause damage to the cell. This is due to a couple of reasons. 1) the photosystems cannot process more than a certain number of photons due to a limited supply of electron carriers. 2) Excess energy can form reactive oxygen species, which cause all sorts of damage. It should be stated that cells are capable of adapting to a variety of intensities in time. Extending the photoperiod under excessive light intensity will cause more harm than good.

I am not saying you should illuminate the cultures with excessive light intensity. That doesn't make sense. I am saying to maximize productivity, cultures should be illuminated at a non-damaging light intensity 24/7.

I have not read this book you are referring to. For my everyday reference, I use "Handbook of microalgae culture" as well as "Algal Culturing Techniques". I would want to read the full passage you are referring to before commenting. You have severely misunderstood another reference you have cited, so I am honestly highly sceptical of your interpretation of this book as well. The concept of 24/7 light for algae cultures is not a debated topic in the field of microalgae biotechnology, so I honestly have no idea why the authors would say that, and I assume there is more to the story. The fact that they refer to "color temperature" shows me this is not a highly scientific publication to begin with. In this science, wavelength and spectrum are referred to, not "color temperature".

I had assumed you were talking about consuming oxygen when you said that, so that doesn't change anything.

This is all completely correct.

There are some serious problems with this information.

"Dark phase respiration" - Are these your words or the authors? It seems to be some combination of "Dark reactions" and "Cellular respiration" which are completely different processes. The dark reactions occur during daylight! They cannot occur at night. Cellular respiration occurs all the time. Plants consume oxygen all the time, but fortunately they produce more than they use while illuminated. At night, they only consume it. It sounds like you are referring to cellular respiration in your description, so remember this occurs all the time (not just at night).

Much of the information you are giving is factually incorrect. I have no desire to contact the people you recommend me to as I am confident, if they were well versed in plant physiology, that they would agree. I believe you are mis-interpreting what they are saying.

I too have cultured many different species of micro algae, but that doesn't teach you about their physiology. It teaches you how to inoculate cultures, aerate them, light them, and keep them going, and if you are observant, a bit about their life cycle. It does not teach you that, for example, they have a limited supply of electron carriers, nor when the Calvin cycle reactions occur.

There are a few practical reasons to light algae at 16/8, as Myka has said. It consumes less electricity and it prevents high pH's if you do not aerate with CO2 to keep it down. But there are no physiological reasons for it.

Jake The problem of all your scientific reasearch is that we are not talking about how to culture phyto in a coca cola bottle, we are talking about culture phyto in a container with the light build inside, as the guy who ask for help said mameroo kit! That is why I think Mou talked about algae bloom and the risk of high temperatures. The only thing I see on you is that you just want to get approval from every thing, if you didn't have culture many species of phyto all your information to me is just a repeating book. As every one have the right to make opinion I guess I would not toletared to you try to destoyef the job of some else when you never done it yourself!

so should my culture be pretty dark after 7 10 days

I really don't know why you seem to be taking this personally. Mameroo is obviously an accomplished hobbyist when it comes to culturing phytoplankton. However, that does not mean nobody can correct him and that he is above others recommendations.

I have honestly tried to keep my prior experience out of this so that mameroo and I could debate on equal ground. It isn't fair to say "well I'm far more experienced, therefore I am right". I want to explain to people why 24/7 lighting is optimal for growth at non-damaging light intensities. If you want to know my credentials, send me a PM. It is irrelevant to this conversation.

I will post a picture of my set up on Tuesday, no problem with that (no pictures at home). I have cultured Isochrysis, Schizochytrium, Tetraselmis, cyanobacteria (spirulina), Dunaliella and Chaetoceros. Right now I focus on Pavlova, Chlorella (fw) and Haematococcus (fw) as they are "hot species" in microalgae biotechnology right now. I think I have almost all of them going right now too just to keep stock cultures in case I need them.

You are clearly taking this personally for some reason. You are not contributing anything to this conversation and are randomly (and falsely) accusing me of trying to "destroy" mameroo and of having never cultured phytoplankton before. So please excuse me, I am going to save my time to reply to people who wish to discuss phytoplankton culture.

Mr. I am not accusing you of anything, I just like to take information and facts. The reason why I do take this personality is because I see the work that this guy have done, now every one comment and criticism...but a year and a half ago not one got the idea to study more about species and bring it in Canada, sorry to take it on you, but I am sick to people saying that they want to help others and turn conversation in to competition for more knowlodge or best answer...Doesn't seems fear to me, as some people like to correct I like to do the stuff right, I respect people and I respect people work. I also think if everyone like the same and do the same we wolud lost our diversity, that i why I love people making opinion but always respecting the other people views. I don't think to make my points I have to step on other people ideologies or sources, we are not in a dictatorship system where everyone have to do what other says. You and him come up with information of different sources, but I keep reading how you try to make all his information wrong and this and that, for me and most of the people in here is very simply for you it works in a 24/7 illumination and for him in 16 hrs. on and 8 hrs. off, if what you suggested or he suggested works for you guys or for the people who wants to follow it and get they results is every one decision. I support what Mou says as I saw his system and I see how well it does, but from with all respect I just see you take information from a book and does it all, I don't see your system with all respect, I think if I walk in a store and they try to sell me something that looks bad, I will buy what I see it looks good, how you see his information is wrong I see it write and I don't see nothing wrong on that. I hope you could share picture and more informations of your system.

I will love if you could provided me some of your cultures, I will like to test the nutritions of your source and mameroo, maybe if I saw you providing live phytoplankton a year and a half ago I will have a different ideology of your suggestion. At this point I support the person who have been making my live stock happy who is Mou.

:popcorn:

From my experienced and what I have learned from Mou it takes about those days for first time, you will be able to harvest between 7 to 10 days. Usually Mou is not a canreef person, you get information from here and if you want information from him I guess the best is to call him during the weekends, e-mail him or I can send you a pm with a web page where you can see his systems and ask questions about the phyto, most people in there cultured they own, some just buy it ready...I am very sure you can get more help and less confusing debating ;)

lol, this is the best comment I saw today hahaha

Canada - phytoplankton free until 2011.

Mr. Jake you like to debating, my question are: are you looking for approval?Do you like to educated people and informed them? Are you a person with open mind to accept other ideologies then yours? I don't like to debating just to accumulated scores ;)...Now the problem with open topics is that they open a thousand conversation, I meant Phytoplankton species to culture, no the phyto what we see on the ocean, lakes or ponds...No the comcentrated phyto what the stores sells...Are we talking about the same phyto, if it is you could give me some sources where they sell it in Canada since 2011, maybe you are right and the stores didn't make enough advertising on selling phyto toculture in Canada, maybe my computer browser don't look for what I type in.

whats wrong with people i thought my post was kinda to the point didn't want anybody getting all up in arms about

I apologize Donkey. I thought I'd answer your question and provide some background on the scientific reasoning behind my answer, but some took offense to the manner in which I did it. If you have any other questions, I'd love to help. I find phytoplankton culture and plant physiology fascinating, and truly enjoy talking about it and helping others learn.

I doubt Mameroo and I would disagree about much other than photoperiod anyhow. Phytoplankton culture on a hobbyist level is very simple once you get the hang of it.

Sorry donkey77, I guess every one have different temperaments, some people try to provided help, other ones make a humility opinion and the rest just try to make all the opinions wrong. I guess I ask ones for help in this page and I add a message to make it clear I just was looking for opinions. I am off now from here, I hope you can get the help you need. Jake I am off, I have not interesting to know if you are right or wrong lol, but I am very interesting to look at your culture system and info on Tuesday, I also hope you can provided the places where people can get phyto species to culture. Thank you, bye!

Well I must I say I have learned a ton from this thread. I have never cultured phytoplankton, but now have absorbed information up to the capacity of my brain. Jake, thanks for the primer on bio-chem. :)

- Ian

all is good my friends
I've learned a lot from this thread so thank you all again and remember the bigger picture
we love this hobby

Thanks! That makes it worth the effort that went into it.

Agreed.
Both Jakegr's and mameroo2000's input is appreciated.