Phyto culturing.Why,Which and How.

[Luis A M]

I enjoy culturing phyto and I find it straightforward. I have been doing this for years and I rarely lose an algal strain. I wanted to share my methods with people willing to give algal culture a try.

WHY
This used to be simple to answer;larval fish feed on zooplankton, and zooplankton feeds on phytoplankton. So anybody attempting to raise fish larvae needs to master the first two levels of the food chain, .i.e. to rear clownfish, you need to culture phyto to feed the rotifers that you offer to the clown larvae.
Microscopic algae are also the perfect food for growing Artemia to adulthood, and for filter feeding invertebrates.
Phyto is also needed for the "green water technique", a popular tool in marine culturists' hands, where the larval water is kept a light shade of green by the periodic addition of algae.
But with the recent advent of frozen algae pastes,it is not clear if there remains any actual reason why we should keep culturing live algae.
A typical clownfish breeder, for instance, can perfectly grow rotifers on paste and even use it as "green water" in the larval tanks. This is unexpected, as dead algal cells can not consume ammonia or CO2, one of the hypotheses on how green water works. It actually contributes to the build-up of both molecules thru the cells break-down.
IMO, the need of live algae remains for culturing more delicate larvae and zooplankters,such as calanoid copepods.

WHICH
The two most popular, easier to culture, and useful are: Nannochloropsis (NAN), aka Japanese Chlorella, not to be confused whith the similar looking and named (but unrelated) Nannochloris, and the Tahitian strain of Isochrysis (T-ISO). Third in the ranking would be Tetraselmis (TET).
NAN is the best alga for rotifers and high in EPA; T-ISO is high in DHA. So a blend of both in different proportions is usually advised for larval rearing. TET is also nutritionally sound and is said to have antibacterial functions.
I culture other species, such as Pavlova, Rhodomonas, and the diatom Chaetoceros for special applications, but they are more difficult to keep.

HOW
I culture my algae under natural light. In winter I keep them in a clear roof deck room, formerly used to grow orchids. Minimum temperatures are set at 13º or 18ºC, depending on the needs, and mantained with a gas heater.
In summer it gets too warm in there and I set the cultures outside, protected from the rain and direct sun.
I use freshly mixed ASW, at 1.010 SG, fertilized with f/2 from FAF. Silicate is added when diatoms are cultured. I keep stock cultures in 100 ml Ehrlenmayer flasks,and according to needs, I upgrade into 2L soda bottles or 10 L bags.
Flasks
Stock cultures are kept pure and possibly sterile in flasks. Each flask is filled with 60 ml of medium, cotton stopped and sterilized in the microwave oven. When cool, 10 ml of a thriving culture is seeded with a sterile pipette.
I keep three flasks for each algal species; old, medium and new. I seed new cultures 2-3 times per month, so this is not too much work. Every morning I agitate the cultures by swirling.
I need to keep stock cultures because it is very difficult to obtain them here. In places where they can be easily replaced when lost, one can do without them.
Bottles.
2L clear soda bottles are seeded with algae from another bottle or from one of the stock flasks.Then it is filled up with medium sterilized overnight with bleach, 1.5ml/6L and dechlorinated with thiosulphate. One small dedicated air pump is used for each bottle, with a rigid airline, so that bubbling is fairly strong.
A bottle reachs peak growth in 10-14 days and it is then harvested.
Bags.
When larger volumes are needed, I use 10L plastic bags,hanging from a PVC structure as shown in FAF catalog. Otherwise use of them is similar to the bottles.

[KathyL]

Very nice write-up.

I would like to add that i like to sterilize the bottle as well as the water. I prefill my bottles 2/3 the way up, add bleach, cap and shake, and let sit overnight. The next day or the next month, as needed, I can dechlorinate, fertilize, and seed with dark green or brown culture.

[Luis A M]

Thanks Kathy,please edit the grammar,we don´t have yet a speller!
Forgot to say that bottles and bags are used once and disposed,took it for granted!

[KathyL]

didn't find much to correct. Mostly add spaces after commas, 2 spaces after periods.

You do English well, much better than my Spanish!

[spk]

Folks,

Fantastic, any chance of getting some pictures of this lot.

I use 5l water containers for making water at 1.017 SG ahead of the time and they will sit for nearly a month before I use them. I have about 12 of these prepared in advance. I use RO water for these.

I use 2l pop bottles but do not throw away unless I have too. If there is a build up of algae on the sides they do a second run.

I have a large air pump running all the bottles that I keep, about 30 of them all going. They sit in the fish room with grolux tube running 24x7

I use approximately three per day. 2 bottles to feed and one to restart cultures. My tunraround time for a bottle is approximately 7-8 days.
I feed 1x5 gallon tub and 4 1 gallon bottles of rots, just started on the brine shrimp and experimenting.

I am looking at adding a carbon based airfilter post pump.

Pictures to follow.

[Amie]

Where is the best place to order T-ISO from?

[Greshamh]

Where is the best place to order T-ISO from?

The best, or cheapest? The best isn't the cheapest, that's for sure.

http://ccmp.bigelow.org/

The Provasoli-Guillard National Center for Culture of Marine Phytoplankton AKA "Bigelow" AKA "CCMP"

I hear good things about the starter cultures from http://www.algagen.com/ but I have no experience with them, or any other starter cultures

Some other places to look would be

http://www.florida-aqua-farms.com/

http://aquaticeco.com/

[Amie]

Great, that gives me some places to start. Thank you !

[Greshamh]

Oops, I shouldn't have forgotten to mention Dan who happens to be a member on this site

http://www.seahorsesource.com/cgi-bin/s ... ods-Algaes

[mpedersen]

I can vouch for Dan's stuff...just got 3 more starters from him this week. Another good source for starters, and I tried T-Iso discs from FAF 4 times, never got one to start, but T-Iso as a liquid from Dan started up and has been great ever since...

Of course, everything depends on what "grade" of purity you want from your T-Iso as well..that's why there's so many different sources for algae and why the prices vary so much.

FWIW,

Matt

[Luis A M]

I have received many of my stock cultures from a Uni collection,ran by a well known phycologyst.This way I can be sure I have the right ID and that a good hardy strain to start by was selected.I try to keep them sterile.

I think people selling algae like those mentioned,also acquired their first stock from scientific collections as well.

I don´t know now,but years ago,FAF discs were very unreliable,I got a TET
culture labeled as something else,and even two species discs,the allien being Chlorella.

[BaboonScience]

And Matt's stocks are clean as well. First tIso that I've had success with in awhile.
Thanks Matt.

[fin farm]

How about some reasons and solutions for cultures failing. For example, I often have a culture (nano) that seems to grow well for a few days and then start to yellow and clump on the bottom. Once it starts to yellow it seems like it is game over for that culture. Or sometimes it seems the nano just won't get a nice green. I don't seem to have the same problems with tetra. I use bleach sterilized water at 1.017 and f2. I'm not sure if it is a temp, lighting or contamination issue.

[DThom]

I have had the same problem with nano. It seems like a bacterial problem or something. Seems to clump, and get slimmy. However my T-Iso is doing great! Still working on it...

I have gotten starter cultures from AlgaGen and they have been great! They also have S strain rot's.

[Tomoko Schum]

I have to make one correction to your write up.

Japanese chlorella is nanochloropsus and not nannochloropsis.
Nannochloropsis is from brackish rock pool (according to Dr. Chuck Amsler, Professor, Department of Biology, University of Alabama Birmingham) and it does not live in fresh water while nanochloropsus does well both in fresh water and salt water.

Tomoko

[Bandeng]

Tomoko

I have inquired about your post on "Japanese chlorella" from National Research Institute of Fisheries Science, Fisheries Research Agency, Japan.

The answer was that the Japanese chlorella cultured in Japan is Nannnochloropsis oculata and there is no genus name nanochloropsus on microalgae at present.

Bandeng

[Luis A M]

Bandeng is right,Tomoko
Nanochloropsus is nothing but misspelled Nannochloropsis!

[mpedersen]

Tomoki, perhaps you meant to differentiate between Nannochloris and Nannochloropsis?

[JimWelsh]

Can anybody post a table showing optimum SG, Temperature, Lighting, Media strength, etc., for the more "finnicky" algaes, such as Pavlova, Rhodomonas, etc.? Any specific notes about the pecularities of the more difficult species would be very helpful. TIA!

[hawklover]

This is some great info! I would also appreciate a table with the optimum growth temps, sg's, all those picky little details.

[hawklover]

Also, would it be possible to get an equipment list together for a setup to be able to mass produce? That would be a great help!

[DaveA]

making water at 1.017 SG

I have two questions:

1. Luis put some nice detailed directions, but I am still left wondering
_ _ a Is this easy or not?
_ _ b How much time does it take for setup of all the equipment?
_ _ c How much time does it take for daily or routine maintenance?
2. Is 1.017 the optimum salinity?
3. You posted minimum temp. Is there a maximum temp?

Can anybody post a table showing optimum SG, Temperature, Lighting, Media strength, etc.

+1 for a table for ALL species. Perhaps we can just start making a table with anyone putting in any factors you personally know

Thanks again! Great info!

[Scottt]

There are tables like that in 'The Plankton Culture Manual' by Hoff and Snell www.florida-aqua-farms.com

Most of the time is in setup. Once you have a culture station, maintenance is minimal. I spend a coupe hours once a week restarting cultures (sterilization is an hour of it). The more rotifers you need to grow, the more time it would take though. IMHO its completely worth it. Live algae cleans your larval water, how can you beat that?

[Luis A M]

There are tables like that in 'The Plankton Culture Manual' by Hoff and Snell http://www.florida-aqua-farms.com

Right, books and algae sites show these data.
I culture at 1:010.Swirling the stock flasks takes 1 minute.
Mass producing requires some 20 minutes/day.
Except for CHAE,they grow well at 13°.About 40°will kill them.
And yes,it is easy.at least for me!

[JimWelsh]

There are tables like that in 'The Plankton Culture Manual' by Hoff and Snell http://www.florida-aqua-farms.com

Right, books and algae sites show these data.

I own and have read "The Plankton Culture Manual" by Hoff and Snell. The tables in it are incomplete, vague, and even contradictory (Temp for I. galbana stated as 25-30 C on page 30, then on page 31 it says 14-22 C, for example). My original question was about less commonly cultivated and "finnicky" species. Others on this site have suggested that T-Iso has more restrictive temperature and light requirements. The diatom R. lens is not mentioned in the tables found in Hoff & Snell at all.

As far as referring the reader to "algae sites", Luis, please throw us newbies a bone, and perhaps provide a link or two! I can't speak for others, but I have done what research I can on the internet already before coming to this site. My specific interest originally was to cultivate the diatom R. lens, so I could attempt the protocol I found in "A Guide to the Meso-Scale Production of the Copepod Acartia Tonsa". My first culture of R. lens immediately crashed -- and I've not ordered another because I still don't have any idea what went wrong. Perhaps there is no such thing as a "finnicky" algae species, and all can be cultivated equaly easily at a wide temperature range and light intensity levels. Maybe I'm making much ado about nothing.....

If there is one place on the internet one might expect to be able to amass the collective knowledge and experience of those who successfully cultivate various phyto species, this just might be it. Why else does this forum exist?!? Referring hawklover and me to "books and algae sites" comes across as a bit dismissive.

I still submit that it would be useful and helpful to attempt to build such a table, and would be willing to work on coordinating such a project. I'm also willing to be convinced that such a table would be pointless and unnecessary, if more experienced members will maintain that all algae and diatom species are equally easily maintained across a broad range of temperature, light, salinity, and nutrient levels.