Phytoplankton Lighting

[Andrew17030]

I know the obvious answer to what light spectrum should be used but this is too simple and likely under thought. If anyone culturing different species could simply provide the species, light source, intensity, spectrum, and approximate growth rate, it might help others. If you have tried different lighting and found one that works better that would be helpful also. I have been looking for research that comparing the effects of different light spectrums or proportions of red/blue and found nothing.

 

[billsreef]

Cool white florescent bulbs will work decently, though I find better results using high CRI full spectrum bulbs. Bulbs set up to light up the full length of the culture vessel do well, this can be a multiple bulbs set up along a bench with multiple culture vessels...aka a two bulb shop light will work well for bench of 2L bottles and a 4 bulb set up will work well with 5 gallon carboys. Larger commercial size vessels will utilize a strip of lights lengthwise for each vessel.

 

[sunny d polyp]

Lighting for Phyto...

Lighting for Phyto...

hey Andrew... I use full spectrum T-8's 4 (4 ft) 24-7 and culture nano in 5/gal card boys and use to feed 10/gal's of roti's. I've had the same rotifer culture for 4 years and usually restart my phyto cultures every 6 months, so I've had good luck with this set up. My card boys last about 10 to 12 weeks and that's taking out 2 1/2 gals every 8 or so days and replacing with sterilized culture water.

Hope this helps.

 

[radiata]

hey Andrew... I use full spectrum T-8's 4 (4 ft) 24-7 and culture nano in 5/gal card boys and use to feed 10/gal's of roti's. I've had the same rotifer culture for 4 years and usually restart my phyto cultures every 6 months, so I've had good luck with this set up. My card boys last about 10 to 12 weeks and that's taking out 2 1/2 gals every 8 or so days and replacing with sterilized culture water.

Hope this helps.

Sunny D,

Are you culturing Nannochloropsis, or something more exotic?

Thanks,
Bob

 

[sunny d polyp]

Phyto culture...

Phyto culture...

hey Bob... nope just nano, I use to culture tet, nano and T Iso G but found that I have less undesired contaminates with just nano and the cultures last much longer. I know some people use special rotifer feeds that have all these extra added nutrients but nano is actually very nutritious on it's own and I'm not feeding larval fish with it. I consider these endeavors to be just another aspect of the hobby and it helps with my OCD.

Jerry S.

 

[Andrew17030]

Awesome, sounds like there are people here that could help me out. Has anyone used containers that buffer PH? They are mostly used by home brewers.

 

[Andrew17030]

http://s938.beta.photobucket.com/user/Andrew_57/media/IMG_1227_zps887a9dad.mp4.html

 

[billsreef]

Awesome, sounds like there are people here that could help me out. Has anyone used containers that buffer PH? They are mostly used by home brewers.

There are specific buffers akin the buffers on the market for aquariums that are sold for adding to the water for homebrewing, but no containers that buffer pH that I've ever heard of.


For my phtyo cultures I just use either natural SW or a well buffered SW mix. More than adequate for typical culturing.

 

[sunny d polyp]

phyto...

phyto...

There are specific buffers akin the buffers on the market for aquariums that are sold for adding to the water for homebrewing, but no containers that buffer pH that I've ever heard of.


For my phtyo cultures I just use either natural SW or a well buffered SW mix. More than adequate for typical culturing.

Same here... I use water from partials that are diluted to 1.020 with r/o and run them thru a 10 micron sieve and then inoculate with .4 ml of sodium hypo.

 

[Andrew17030]

Ignore post #7, wrong thread and doesn't seem to be connecting anyway.

Yea, I found some containers that were alleged to buffer pH themselves accidentally and thought they could be helpful. I haven't found them again at this point. It's possible, not sure how it well would work when not air tight though. I'll keep looking.

Billsreef, what shape bottles do you use? I was thinking about using boston rounds, wide mouth, sloped shoulder, flat bottom, 4-32oz., and about 1.5 dollars glass or PET.

Have you used air filters inline with your air supply? I'm having a hard time finding anything smaller that 5 microns or anything used specifically for air at that size. I found an ice machine water filter with a simple design that could work at 5 microns.

 

[billsreef]

Sure those containers weren't the "Better Bottles", which are simply lightweight oxygen impervious PET carboys used for fermentation instead of the heavy glass carboys? With brewing beer, it's the mash where pH counts, and I don't see any type of container having an influence on the mash pH...which is also open to air, so air exposure would be irrelevant.

For culture vessels, I like Erlenmeyer flasks for small subcultures and step up cultures. Using a foam stopper in the flask, you can sterilize the culture media and the flask all in one shot using a microwave or autoclave (if you got one of those ). For larger working cultures, 2L soda bottles works for small scale home based work, 5 gallon carboys for larger scale work...and the sizes go up from there for larger scale commercial and research work.

By way of air filters, Florida Aqua Farms sells a nifty little inline airfilter that you can stuff with filter floss and change out periodically. I find it works well.

 

[Andrew17030]

Nope they were Borosilicate with some forgotten coating and they were only one L.

No autoclave, microwave is doable though. What about chemical sterilization? I have read lots of info where muriatic acid was diluted and used to sterilize vessels and equipment, why acid and not chlorine?

I see Erlenmeyers used a lot. Just thinking it through but the shape of the Erlenmeyer would not be as efficiently lit, aerated, and mixed as a more narrow cylindrical container.

I want to culture T-Iso (Isochrysis Galbana), TW (Thalassiosira Wessfloggi) , and RL (Rhodomonas Lens). Have any experience with TW or RL?

 

[Andrew17030]

Bad memory, not used for brewing, here they are:
http://www.freundcontainer.com/type-graduated-media-glass-bottles/p/V1010B01/

 

[billsreef]

Ahh, those bottles don't buffer, they just don't react with what is in them or allow any sort of atmospheric gas exchange

The muriatic acid isn't used for sterilization, but rather as a final rinse to remove any deposits from things like diatoms that might get in the culture. Keeps the culture vessel squeaky clean. Bleach is still the desired option for chemical sterilization

The Thalassiosira is going to be the easiest of that lot to culture. Maintaining a really clean set of subcultures is the best way to maintain and be constantly starting new batches in order to keep those strains going well. I think you'll find doing batch cultures (with very good sanitary technique) will give the best long term results. If you haven't already, get yourself a copy of Frank Hoff's Plankton Culture Manual. It is an excellent source of culturing info, pretty much considered the bible for culturing phyto, rots, etc.

 

[Andrew17030]

I wanted to do progressive batch, 32oz. to 64oz. to 3g. to 6g. ending up at a 6 gallon carboy. I am thinking about starting with small containers so that I will not need to thin small sensitive cultures and add media as needed. How do pros move a culture from one bottle the the next size? I can't think of anything that would gentle and relatively sterile except maybe a metering pump. Seems like a big waste of time though.

 

[billsreef]

The way it typically gets done (this is also how I've done it at the lab) is to maintain 3 sub cultures for every large batch you want to start. This allows for contamination issues or crashes with enough redundancy that you don't loose the culture line. The subcultures are typically small, around 50 to 75mls...I like 100 or 150ml flasks for this, with foam stoppers. These subcultures are small enough that simple given them a swirl once a day is adequate. One set of subcultures is ready to step up, use one to inoculate a 2L flask (generally about 1.5L worth of culture). The 2L flask is put on air, and generally ready to step in a week. Once that 2L culture has gotten to peak density, transfer it to the Carboy....never found any real need for an intermediary step up between the 2L and Carboy. Do all your transfers in a clean area, with clean equipment, and that will be sterile enough for the step ups. Also, when you do that first inoculation from the subcultures, use one of the 3 subcultures to start the next set of subcultures...if things have gone well you have a spare subculture that you can use to start an extra box, or toss if not needed. It's the subculture part that is the most critical for sanitation, as this is your clean culture source While some people will often simply start new batches from an existing batch, it typically only works well for robust species like nano...the ones your looking at crash to easily for that approach to work well, though they are excellent nutritionally and worth the effort IMO.

 

[sunny d polyp]

smaller bigger biggest...Hoipe this works?

smaller bigger biggest...Hoipe this works?

smaller;
http://s1104.beta.photobucket.com/user/sunnydpolyp/media/Picture820_zps197e2ac7.jpg.html?sort=3&o=5

bigger;
http://s1104.beta.photobucket.com/user/sunnydpolyp/media/Picture121_zps35ec53c0.jpg.html?sort=3&o=3

biggest;
http://s1104.beta.photobucket.com/user/sunnydpolyp/media/Picture005_zpsab2c85f2.jpg.html?sort=3&o=2
http://s1104.beta.photobucket.com/user/sunnydpolyp/media/Picture006_zpsfffd2637.jpg.html?sort=3&o=1

rotifer station
http://s1104.beta.photobucket.com/user/sunnydpolyp/media/Picture_zps8a2d8c71.jpg.html?sort=3&o=4

 

[Andrew17030]

Billsreef, thanks. I have a tendency to over-complicate simple things that I think I can improve upon. What I am finding out is that I suck at keeping things simple.

Okay, I'm gonna reiterate what I think you are saying.

Use three container sizes approximately (small:50-150ml, medium:1500-2000ml, large:5 gallon carboy) and manage subcultures and cultures/batches independently.

I want to clarify what I think batch means. A batch is a given amount/container(s) of cultured algae progressing from one vessel to the next.

Maintain three subcultures in small containers without aeration per each medium aerated culture that I wish to inoculate. Additionally, use only subcultures to start subcultures. And then, one medium culture is enough to start one large culture when the cell density in the medium sized vessel reaches the end of logarithmic growth. Then grow out the large culture and use when desired density is reached. I have a question. Would it be a good idea to begin that 1.5 L culture in a total of one gallon of water and add medium and buffer as needed to maintain logarithmic growth as opposed to a different amount?

Are there simple ways to test pH and density that are not overly complicated and worth the risk if done right? Is this overly complicated to even consider? I'm not doing research. I just want to feed this stuff. Maybe handoff to someone that needs a start up, which would be done from my subcultures, right?

I've been in chemistry labs for school but it was hard to mess the stuff we did up because of microbe contamination. What would be an appropriate setup to perform inoculation, sanitization, and transfers? I am assuming relatively clean air, I have a ozone air purifier in my fish room which is a small guesthouse about 1000 square feet.

Here is a picture of my culture station. So far I have six dual T12s and a few CFLs up top for my smaller cultures that I don't want to grow very much. I was thinking that turing these smaller lights on first and stagger the rest would help with potential photo-shock. The rack is a metro shelf that is often used in restaurant kitchens. It's been cleaned and sanitized. The shelves are adjustable but its kind of a hassle and putting one together in a closet is not recommended. Yes it's a closet. I know that temp may become a problem especially with RL. I may put a small ac unit in there run off my aquacontroller. I wanted to use a closet to help prevent contamination. Where I am stumped with this idea is heat control, exhaust fans vs. small AC unit vs. simple fans. I am not sure fans will do anything except move the heat more evenly and exhaust fans will create negative pressure and air will enter from the surrounding room anyway limiting my control over temp and contaminants.

<a href="http://s938.beta.photobucket.com/user/Andrew_57/media/IMG_1697_zps15c4d83a.jpg.html" target="_blank"><img src="http://i938.photobucket.com/albums/ad225/Andrew_57/IMG_1697_zps15c4d83a.jpg" border="0" alt="Photobucket"/></a>


Any ideas?

 

[Andrew17030]

Sunny d, are those large run continuously? I would think that keeping those spiqots from causing fouling would be tricky.

Safari could not keep them open if anyone is trying to link to those files firefox worked for me.

 

[sunny d polyp]

cultures...

cultures...

Yes sir, I run them 24/7 and have the card boys labeled A B C D and a whiteboard to keep up with the Algro additions and the partials (2 1/2/gal every 4 or 5 days). I use filtered air (FAF) and inoculate with .4 ml's per 1/gal tank partials. I have had "A" running since 10-28 and it is still getting very dense. My cultures naturally stay at a higher than normal PH and I don't worry about about it, it's never caused a problem.

I think about what I'm about to do before I make a move and ALWAYS keep my hands sterile with hand sanitizer. When I remove the 2 1/2 gal partial I wipe the inside (dry part) of the card boy clean with a paper towel dipped in the sterilized culture water before I use the sodium thio sulfate to neutralize the chlorine and remove the lid and give it a bleach water bath. I keep the outside of the container wiped down with the sanitizer as well. You can't be too careful with the cleanliness because if you do everything else just a bit wrong you can get away with it (reasonably) but bacteria/fungus are your enemy.

The bags were a pain and every 8 days (exponential phase) I would dump, sieve, inoculate and pour the culture into new bags, got very old (17 liter bags are appx 40 lbs full of phyto).

This is the simplest solution I've came up with and so far is great.

If you are interested in this set up I can get more detailed information and send better pics also explain in greater detail.