Culturing and feeding Phytoplankton to Coral, Pods and Fish

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Hi Brad

Been following for a while, looking great. Enjoyed the evolution

2 things, though

1) unfortunately, I think you have tetraselmis, and not Nanno. The Tet is a better strain of phyto (sifter fell wall compared to nanno and better for pods) but it is harder to culture. Your success is better than initially appreciated

2) what’s the purpose of the 51 micron filter? Is it just to filter out clumps and ‘mulm’ (broken down cells that make that brown sediment in the bottom)? I only ask because I use one to separate out adult from juvenile pods in my T. californicus cultures

Thanks man, looking great [emoji106]
 

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As a comparison, here is a sample of my newly split, 2 month old (so, 6 splits at this point from inoculation) nannochloropsis, taken at 400x (but cropped, so actual magnification unknown)

e0e204961aee2a9406ad64167f659831.jpg
 
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Brad Miller

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Hi Neil,
Your right...my culture should have been Nanno, but I was corrected just a bit ago on a microscope thread.
I used the 53 sieve, expecting to separate the sludge, but am not really getting anything so, I’m not using it anymore.
I’ll post a cool video showing the differences between a few strains below;
 
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Brad Miller

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I sent the videos and pics I took of my culture to Mercer Of Montana.
They just emailed me back and confirmed that it is definitely Tetra !
This is kinda like raising a chicken for months, then finding out that it’s really a rabbit :)
They said their culture rooms are separated and they don’t know how this could have happened...I’m so proud of my microscope :)
 

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Hi Brad...I recently purchased the Phyto and Copepod farms from Poseidon reef systems. They have fantastic customer service and the products work as advertised. Ive done a lot of DIY on my tank but this time I decided to just get a kit and I'm glad I did. I had phyto in a week and have a strong colony of pods several weeks later. I know you can do this all DIY but every now and again I like to just get it done. This system comes with everything you need including cultures, fertilizers, and all the accessories. Very nice product and no cleanup.

20181208_144530.jpg
How much would a kit like that set ye back? Or would I be better served setting it up myself?
 
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This is the kind of information I have been looking for and thought that it would be great to post this here for reference in case it gets harder to find out there.

I found this article from 2002 and even though it is an older one, it spells out the nutritional benefits of each of the Phytoplankton we usually deal with in our reef tanks.
It’s a bit long but well worth it, as it is in favor of feeding Phytoplankton to our systems and explains why.

I am now culturing both Nanno and Tetra for my tank.

If you want to read the complete article with noted references, here it is;
https://www.advancedaquarist.com/2002/8/breeder


Article is copied from the Advanced Aquarist, August 2002, The Breeders Net, Culturing larval fish foods, Part 1;

By Frank Marini, Ph.D.,


When it comes to raising larval fish, nutrition is one of the most critical factors to success. For fish fry, the principal step in the food chain is phytoplankton, and while it is rare that we feed phytoplankton directly to larval fish, we utilize phytoplankton to enrich a food items -- rotifers, copepods, brine shrimp nauplii, etc. These enriched food items are fed directly to larvae. Rotifers, Artemia nauplii, and copepods that are depleted of essential nutrients, have little, if any food valve, and therefore it is critical that we don't ignore the role of providing these nutrients via phytoplankton. Due to the fact that food items take on a similar nutritional value as the phytoplankton cells that they consume, the nutritional value of the phytoplankton is of paramount importance to success with our larval fish.

Phytoplankton are simple, unicellular organisms capable of photosynthesis. Unlike higher plants that are composed of multiple cells and differentiated tissues, phytoplankton lack a stem, any type of roots, or leaves. Because photosynthetic organisms manufacture their own food, they form the basic energy source that sustains many natural food chains. These plants are the starting point.

So What Makes Phytoplankton So Nutritious?
The focal point of nutrients in these microalgaes is the concentrations of omega-3 fatty unsaturated fatty acids (HUFAs). Numerous studies have shown that marine fish are unable to synthesize sufficient quantities of two essential HUFAs; Eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) [Kanazawa, 1979.] These two fatty acids are essential in the growth and development of fish. In general terms, the higher the level of HUFAs, the more nutritious the phytoplankton are to fish.

The Microalgae
Appropriate microalgae that are readily cultivable at home, and are suitable food for prey items which will be fed to fish fry are critical to our home breeding success. There are approximately 7000 species of microalgae, although many of which are not adaptable for home culture. Currently a few species are readily available and readily adaptable to home culture: Nannochloropsis oculata, Chaetoceros gracilis, Isochyrsis galbana, and Tetraselmis sp_._ The nutritional value of each of these microalgae vary, which makes some species more appropriate for our use than others. (Table 1). In the following paragraphs I will describe several useful phytoplankton species.

Table 1 Comparison of phytoplankton discussed in the text
Algae Total HUFA EPA DHA
N. Oculata 16-43% High Low
C. Gracilis 5-11.5%. 0.3-2.5%
I. Galbana 2-4% 3-4.2%
T. Iso 0.2-0.7% 8.3-11%
Tetraselmis ~5% ~6%

Nannochloropsis oculata is a 2-4 micron (μm) green flagellate. This is a fast growing species that is easy to maintain. This phytoplankton is the one most commonly thought of when the term green-water is used. This is a dark green alga with a thick tough cell wall that interestingly is readily consumed by rotifers. N. Oculata is high in overall omega-3 HUFAs (ranging from 16-42%), and while most of the HUFAs are composed of EPA, there is little DHA present. A growth study performed by Okauchi et al [Okauchi 1990] determined that the highest level of EPA was attained at 7 days after batch cultures were inoculated. N. oculata has been shown to contain very high levels of vitamin B12, which is critical for larval fish survival, and it has also been suggested that vitamin B12 is important for developing diseases resistance in larval fish as well.

Chaetocerous gracilis is a 6-9 μm solitary diatom with four large spines. It is frequently used in large quantities in commercial shrimp culturing. Because of its protruding spines it has been suggested that this phytoplankton can be problematic in rearing food items; however, this problem has never borne out in commercial cultures. C gracilis has an EPA range from 5-11% EPA and DHA from 0.4-2.5%.

Isochrysis galbana is a 4-7 μm golden-brown flagellate. This species is commonly used in bivalve culture (clams, oysters, etc). While it has been occasionally used as a single rotifer food, it is usually mixed with other phytoplankton such as chlorella or N. ocultus. The EPA levels range from 2-3.5% and DHA is 3.5-4%. Different strains of this species have varying levels of HUFAs, and one isolate found off Tahiti (commonly known as T-Iso) contains high DHA (8-11%) and low EPA (0.2-0.7%). This EPA level is much lower than found in a standard reference strain of I.galbana. An important note for home culture is that this strain requires consistent temperatures, vitamin additives to the nutrient broths and a silicate additive to reach maximum density. According to Wilkerson [Wilkerson, 1998] this algae is too temperamental, fragile, and fastidious to be used regularly.

Tetraselmis sp. is a 9-14 μm motile green flagellate, which has been successfully used in outdoor ponds because it is extremely temperature tolerant. There are several species of Tetraselmis sp. that are available and one such T. tetrathele is frequently used in aquaculture. Studies have shown that while EPA (~ 5%) and DHA (~7%) levels in this phyto are theoretically sufficient, several authors has suggested that rotifers feed diets exclusively on T. tetrahele were not capable of sustaining fish larvae [Fukusho 1985, Wilkerson 1998]. To combat this deficit, aqua-culturists have fed mixtures of T. tetrahele with other phytoplankton species and discovered that these combinations were significantly more nutritious than those cultured alone. Of interest to hobbyists, Tetraselmissp, produces two antibiotic- like compounds which have been documented to increase survival in larval fish feed on prey items enriched with this phytoplankton.

Given the above information on the available phytoplankton, which one is the best to use? Considering primarily ease of growth and sufficient HUFA profiles, N. oculata is my first choice, followed by C.gracilis and I. galbana, and lastly T-iso. While C. gracilsis and I. galbana has similar nutritional profiles, C. gracilis grows more rapidly and more consistently in culture. Another important consideration is which of these phytoplankton will grow under home water conditions. Each phytoplankton species and strain has an optimum pH and salinity range in which it grows best. It is only through experimentation that you will discover which one grows best for you.
A chart of "optimal parameters" is provided to allow you to make some comparison {Table 2} [Wilkerson 1998 pg157].

Table 2: Optimal conditions for phytoplankton discussed in the text
Algae Optimal pH Temp Range Minimum Illumination (LUX) Salinity
N. Oculata 7.0-8.4 60-86 4,000-5,000 22-25
I. Galbana 7.8-8.5 77-86 1,000-6,000 28
Tetraselmis 6.9 68-82 1,000-20,000 30-40

According to commercial experts, rarely is the use of a single phytoplankton suitable for aquaculture of fish larvae. Nutritional deficiencies found in one phytoplankton species can be compensated for by adding another phytoplankton species superior in that missing HUFA. As an example; N. oculata which is high in EPA, but low in DHA can be paired with T-Iso, which is high in DHA. Some hobbyists even add a small portion of Tetraselmis to this co-culture just to add an antibiotic effect. Studies performed in commercial fisheries have shown that fish larvae fed prey items enriched on diets composed of multiple phytoplankton species have higher survival rates and quicker growth rates than those larvae fed food items enriched with a single type of phytoplankton.

The "take home" message here is that the use of multiple phytoplankton species is advantageous.
If you must only use a single culture of phytoplankton use the one with the highest HUFA concentrations.
 
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Brad Miller

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You would culture the Phyto and feed it to your Rotifers, pods, etc.. by dumping it into their tanks or culture containers or buckets, keeping the color of the container a light green so you know they have food in there.
 

Reefthedayaway

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D
You would culture the Phyto and feed it to your Rotifers, pods, etc.. by dumping it into their tanks or culture containers or buckets, keeping the color of the container a light green so you know they have food in there.
Do you have a set up like that?
 

sde1500

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This is the kind of information I have been looking for and thought that it would be great to post this here for reference in case it gets harder to find out there.

I found this article from 2002 and even though it is an older one, it spells out the nutritional benefits of each of the Phytoplankton we usually deal with in our reef tanks.
It’s a bit long but well worth it, as it is in favor of feeding Phytoplankton to outer systems and explains why.

I am now culturing both Nanno and Tetra for my tank.

If you want to read the complete article with noted references, here it is;
https://www.advancedaquarist.com/2002/8/breeder


Article is copied from the Advanced Aquarist, August 2002, The Breeders Net, Culturing larval fish foods, Part 1;

By Frank Marini, Ph.D.,


When it comes to raising larval fish, nutrition is one of the most critical factors to success. For fish fry, the principal step in the food chain is phytoplankton, and while it is rare that we feed phytoplankton directly to larval fish, we utilize phytoplankton to enrich a food items -- rotifers, copepods, brine shrimp nauplii, etc. These enriched food items are fed directly to larvae. Rotifers, Artemia nauplii, and copepods that are depleted of essential nutrients, have little, if any food valve, and therefore it is critical that we don't ignore the role of providing these nutrients via phytoplankton. Due to the fact that food items take on a similar nutritional value as the phytoplankton cells that they consume, the nutritional value of the phytoplankton is of paramount importance to success with our larval fish.

Phytoplankton are simple, unicellular organisms capable of photosynthesis. Unlike higher plants that are composed of multiple cells and differentiated tissues, phytoplankton lack a stem, any type of roots, or leaves. Because photosynthetic organisms manufacture their own food, they form the basic energy source that sustains many natural food chains. These plants are the starting point.

So What Makes Phytoplankton So Nutritious?
The focal point of nutrients in these microalgaes is the concentrations of omega-3 fatty unsaturated fatty acids (HUFAs). Numerous studies have shown that marine fish are unable to synthesize sufficient quantities of two essential HUFAs; Eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) [Kanazawa, 1979.] These two fatty acids are essential in the growth and development of fish. In general terms, the higher the level of HUFAs, the more nutritious the phytoplankton are to fish.

The Microalgae
Appropriate microalgae that are readily cultivable at home, and are suitable food for prey items which will be fed to fish fry are critical to our home breeding success. There are approximately 7000 species of microalgae, although many of which are not adaptable for home culture. Currently a few species are readily available and readily adaptable to home culture: Nannochloropsis oculata, Chaetoceros gracilis, Isochyrsis galbana, and Tetraselmis sp_._ The nutritional value of each of these microalgae vary, which makes some species more appropriate for our use than others. (Table 1). In the following paragraphs I will describe several useful phytoplankton species.

Table 1 Comparison of phytoplankton discussed in the text
Algae Total HUFA EPA DHA
N. Oculata 16-43% High Low
C. Gracilis 5-11.5%. 0.3-2.5%
I. Galbana 2-4% 3-4.2%
T. Iso 0.2-0.7% 8.3-11%
Tetraselmis ~5% ~6%

Nannochloropsis oculata is a 2-4 micron (μm) green flagellate. This is a fast growing species that is easy to maintain. This phytoplankton is the one most commonly thought of when the term green-water is used. This is a dark green alga with a thick tough cell wall that interestingly is readily consumed by rotifers. N. Oculata is high in overall omega-3 HUFAs (ranging from 16-42%), and while most of the HUFAs are composed of EPA, there is little DHA present. A growth study performed by Okauchi et al [Okauchi 1990] determined that the highest level of EPA was attained at 7 days after batch cultures were inoculated. N. oculata has been shown to contain very high levels of vitamin B12, which is critical for larval fish survival, and it has also been suggested that vitamin B12 is important for developing diseases resistance in larval fish as well.

Chaetocerous gracilis is a 6-9 μm solitary diatom with four large spines. It is frequently used in large quantities in commercial shrimp culturing. Because of its protruding spines it has been suggested that this phytoplankton can be problematic in rearing food items; however, this problem has never borne out in commercial cultures. C gracilis has an EPA range from 5-11% EPA and DHA from 0.4-2.5%.

Isochrysis galbana is a 4-7 μm golden-brown flagellate. This species is commonly used in bivalve culture (clams, oysters, etc). While it has been occasionally used as a single rotifer food, it is usually mixed with other phytoplankton such as chlorella or N. ocultus. The EPA levels range from 2-3.5% and DHA is 3.5-4%. Different strains of this species have varying levels of HUFAs, and one isolate found off Tahiti (commonly known as T-Iso) contains high DHA (8-11%) and low EPA (0.2-0.7%). This EPA level is much lower than found in a standard reference strain of I.galbana. An important note for home culture is that this strain requires consistent temperatures, vitamin additives to the nutrient broths and a silicate additive to reach maximum density. According to Wilkerson [Wilkerson, 1998] this algae is too temperamental, fragile, and fastidious to be used regularly.

Tetraselmis sp. is a 9-14 μm motile green flagellate, which has been successfully used in outdoor ponds because it is extremely temperature tolerant. There are several species of Tetraselmis sp. that are available and one such T. tetrathele is frequently used in aquaculture. Studies have shown that while EPA (~ 5%) and DHA (~7%) levels in this phyto are theoretically sufficient, several authors has suggested that rotifers feed diets exclusively on T. tetrahele were not capable of sustaining fish larvae [Fukusho 1985, Wilkerson 1998]. To combat this deficit, aqua-culturists have fed mixtures of T. tetrahele with other phytoplankton species and discovered that these combinations were significantly more nutritious than those cultured alone. Of interest to hobbyists, Tetraselmissp, produces two antibiotic- like compounds which have been documented to increase survival in larval fish feed on prey items enriched with this phytoplankton.

Given the above information on the available phytoplankton, which one is the best to use? Considering primarily ease of growth and sufficient HUFA profiles, N. oculata is my first choice, followed by C.gracilis and I. galbana, and lastly T-iso. While C. gracilsis and I. galbana has similar nutritional profiles, C. gracilis grows more rapidly and more consistently in culture. Another important consideration is which of these phytoplankton will grow under home water conditions. Each phytoplankton species and strain has an optimum pH and salinity range in which it grows best. It is only through experimentation that you will discover which one grows best for you.
A chart of "optimal parameters" is provided to allow you to make some comparison {Table 2} [Wilkerson 1998 pg157].

Table 2: Optimal conditions for phytoplankton discussed in the text
Algae Optimal pH Temp Range Minimum Illumination (LUX) Salinity
N. Oculata 7.0-8.4 60-86 4,000-5,000 22-25
I. Galbana 7.8-8.5 77-86 1,000-6,000 28
Tetraselmis 6.9 68-82 1,000-20,000 30-40

According to commercial experts, rarely is the use of a single phytoplankton suitable for aquaculture of fish larvae. Nutritional deficiencies found in one phytoplankton species can be compensated for by adding another phytoplankton species superior in that missing HUFA. As an example; N. oculata which is high in EPA, but low in DHA can be paired with T-Iso, which is high in DHA. Some hobbyists even add a small portion of Tetraselmis to this co-culture just to add an antibiotic effect. Studies performed in commercial fisheries have shown that fish larvae fed prey items enriched on diets composed of multiple phytoplankton species have higher survival rates and quicker growth rates than those larvae fed food items enriched with a single type of phytoplankton.

The "take home" message here is that the use of multiple phytoplankton species is advantageous.
If you must only use a single culture of phytoplankton use the one with the highest HUFA concentrations.
Their breeders net series is quite a good informative read.

Do we grow phyto then add rotis & copepods? Or do we grow them separately in a different setup?

Rotis will 100% wipe out any phyto culture if kept too close. Be sure to culture them separately.
 

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Would like to see someone's set up. For keeping phyto and rotis/copepods.

Nannochloropsis in the bottle, Tigriopus californicus in the square plastic tub

46a9ab0c4f744318e8759a728bb3205e.jpg
cc08620f1407b42fbf9fdd47d105033d.jpg
8519339a03ab2f0d8893eea1cb6c7a8b.jpg


I’m ramping up production for this new format. I expect I’ll be getting 4-5k adult pods/ week once I get a few more rounds of replication in there. I filter with a 300micron sieve to catch adults and leave everything else.
 

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Water changes are done by siphoning through a 53 micron sieve- which captures everything- dumping the waste water into a bucket of bleach, then flipping the sieve and washing the pods back into the culture vessel with a bottle of nannochloropsis water and then topped up with 1.019 sg saltwater. This is done twice a month currently
 

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Hey everyone.I've been reading through the threads. I started my first Phyto culture seven days ago. Mercer Of Montana kit. I also started a Tisbe pod culture about a month ago. Just did my first big harvest and added them to my sump and some in the tank. I'm going to be following this thread. Great info.
 

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I got the PoseidonReefSystems phyto and pod culture kit a few weeks back and it has already paid for itself. It's plug and play really. I'm only doing 1 strain of phyto and pods. I'd recommend it to anyone wanting to start up culturing. I have more info in my build thread about it if you get curious...
:) same here. I though it was expensive but it has definitely paid for itself, I have also noticed improvement. And like you say, plug and play, even a noob like me can figure it out, definitely recommend their system.
 

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@Dana Riddle
Would you mind posting in this thread on how you do yours ?
I think you would have some great insight for someone looking to get into this.
My approach to microalgae culture is definitely hit or miss. I bought a pint of greenwater from a good LFS, where they have huge colonies of red and green Goniopora specimens. They attribute their success with these to additions of greenwater. I didn't want to spend $40 monthly buying their cultures so I ordered f/2 formula, spiked exchange aquarium water with it and a few milliliters of greenwater, and voila! - 100 gallons of microalgae. My three Gonioporas are growing and the water swarms with zooplankton at night. I also have a Lamarck's angel that constantly picks the live rock for pods. Interestingly, the nitrate concentration has fallen since I started greenwater additions (the skimmer is off during the day and strips out the uneaten microalgae at night.) I've got to get serious in examining the dynamics of greenwater additions. I've received a block heater and total nitrogen/phosphorus reagents from Hach in order to examine import/export. Need to count cells using my microscope and hemocytometer. And use my PAR meter to looking at light intensities over the culture vessels. Wish I had more to present, but I can say I am definitely a greenwater fan.
 

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Do we have anyone who cultures Tisbe Pods? I've been trying for just over a month. The adult size ones I started with seem to be way larger than I'm able to raise. Mine are tiny. So tiny they seem to still make it through a 20 micron filter/sieve. So makes it almost impossible to harvest them and do a water change. One day it seems like I have a ton of them. The next day it's like nothing is in the vessel. Any good tips?
 
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