Post by MADMIKE on Mar 22, 2007 14:01:16 GMT -5
Brine Shrimp Grow Out Article
Reproducible method for brine shrimp grow out - written by Greg Hiller 3-97, revised 6-98
I have used this method over the past two years with very good results. The consumables usage is quite low, and the labor involved is also low, but a small amount of labor is required each day for the best results/highest yields.
What you need
1. As many clear two liter coke bottles as you can get your hands on. Remove the labels. Rinse with tap water. After use it is hard to clean these, but possible if you have to. I just use tap water, hand dishwashing soap, and a brush that can reach to the bottom of the bottle. You can of course use glass bottles, etc. if you want.
2. A pure culture of Nannochloris algae (the disks are the easiest to use,~$7. Revision: I actually think a liquid culture is easier to use now. The nice thing about a disk is that in some cases you may be able to save part of the disk for use in the future (months later) in case your algae culture becomes contaminated. I'm also not really sure which algae strain is the best to use. Most public aquaria use Isochrysis galbana, which I've been told from Carolina Biological is prone to crashing. Also commonly used is Monochrysis, and Carolina tells me this is easier to grow. Although I only grew this one for a short period of time once, it did seem to grow very quickly.
3. Micro Algae Grow
4. Silicone antifoam
5. Brine shrimp eggs (for you purists, Artemia salina cysts), I recommend OSI collected from the Great Salt Lake. I have found these to hatch many timesgreater amounts of shrimp per volume of eggs. The eggs also disperse better in the water, and seem to be much cleaner based on the cloudiness of the water after hatching. I've heard that eggs from collected from the San Francisco Bay are smaller in size at hatching, but if you're interested in growing them then this is not really a concern.
6. A strong air pump (this is most likely going to be the limiting factor for the amount of brine shrimp you can grow)
7. A pair of 40 watt cool-white fluorescent lights (I bought one for $10, the longer the bulbs, the more shrimp you can grow)
8. Flexible airline tubing
9. Rigid airline tubing
10 . Synthetic salthingyer mix
Items 2 through 4 above are available from Florida Aqua Farms (352-567-8540). You can also purchase the algae culture from Carolina Biological, and their cultures tend to be a bit more predictable (1-800-334-5551). To save money on salt water mix the shrimp and algae can be grown in water with a specific gravity of about 1.018. It is very important to maintain at least one micro algae culture under aseptic (sterile) conditions. This can be done by sterilizing the water used to culture the algae. The easiest way at home is to use the microwave. A one liter glass bottle of water can be sterilized in about 7-8 minutes in most microwaves.
Alternatively you could use membrane filtration for sterilization. If you use heat for sterilization, it is best to add the salt mix, but not the micro algae grow (fertilizer)before the sterilization. After the water has cooled, carefully (trying to keep things clean) add the micro algae grow (i.e. fertilizer) to the water in the concentration recommended on the bottle. I have found that 0.6 ml or about 1/8th teasthingy works well for 1 liter of water.
Follow the instructions on the micro algae pure culture that you received to get the algae into suspension. If it is a algae disk place an 1/8th inch layer of water over the surface of the disk and leave in a moderately lit area for a day or so. Rub the algae off the surface with a sterile Q-tip. Pour the suspension into asterile bottle with your fertilized, sterile salt water, place the bottle in front of your fluorescent lights (side as opposed to above lighting is best), and put your bubbler in place. The bubbler runs air from your air pump to the bottle. You can cut the rigid plastic tubing a few inches longer than the depth of the bottle, then using a cigarette lighter heat the tubing and put a 90 degree bend in the tubing so that the bubbling end of the tubing is just slightly off the bottom of the bottle when it is in place. For the bottle with your pure algae culture, try to use a tight fitting cap with a hole in it for the bubbler. You may need to loosen it a bit for the air to escape. The air that runs into your pure culture should probably run through a sterilizing filter before going into your culture.
You can prepare a homemade sterilizing filter by stuffing some filter floss or glass wool into a large diameter piece of tubing. Remember not to add too much floss, or you will not get enough air into your culture. You want the air to really turn the water over in your culture so that the algae stays in solution. The algae culture may take some time to adapt to your water conditions, etc., so at first it may sometimes fall out of solution a bit.
When the algae culture has grown to a point at which you can no longer see through the bottle, you can expand the culture. First prepare a new bottle that will hold your sterile, stock culture. You can typically expand your culture about 3 fold without any difficulty. If you try to expand further than about 7 fold you may risk having the culture be more difficult to keep in suspension, or suffer a very long lag time before the culture comes back up to a high density. After you start the new algae culture, you can use the rest of the algae for your brine shrimp growth.
I don't try to keep the bottles that I use for the brine shrimp growth under sterile conditions, although I do try to keep things clean as much as possible. As long as you carefully maintain a pure culture of the algae, and go back to it on a regular basis to start your expanded cultures for brine shrimp growth, you should not have a problem with contamination. The principal contamination problem that I have incurred is with what I believe was a blue-green algae. Over the course of a few weeks my algae culture seemed to grow faster than I had previously recalled, and grow to a very opaque consistency. Also, the brine shrimp I put into the algae seemed to grow extremely slowly, so much so that there was no point in continuing to attempt to culture them.
[Revision: I have had other types of contaminations: sometimes the algae will form stringy clumps, or diatoms will grow on the sides of the two liter bottles. Fortunately, these types of contamination do not kill or significantly slow the growth of the brine shrimp. The only limiting factor in this case is the problem that the brine shrimp can only eat small particles, and the smaller the brine shrimp the smaller the particles it can consume. FWIW recently I've heard that these stringy clumps are formed by a type of fungi that can grow in the cultures. I've also been told that it can be effectively removed by filtering the culture through a net, the micro algae that you want will flow through]
For the brine shrimp growth you can use several different methods depending upon how much time you want to spend, and how much brine shrimp you want to recover. The first method is to hatch the brine shrimp directly in the bottles you will use for grow out. This means you expand your left over algae culture (after starting a new culture) to a 2 liter bottle, and put a very small pinch of brine shrimp eggs in the bottle. You also need to fertilize the added salt water, and then add a drop, or dab of the silicone antifoam.
[Revision: Keep the antifoam in a sealed container that is opaque to keep out light, things will grow in it otherwise. ]
If you do not use the antifoam, when your cultures start to grow and you get some protein production, a large number of the shrimp will die as they are trapped in the foam layer that forms (this 'aquarium' is a bit small to use a protein skimmer!). Fill your 2 liter bottles only until you reach the area at the top of the bottle where the neck begins to narrow. The trick with this method of culture (as with all the rest I will mention) is that you need to keep the algae culture growing a bit faster than the brine shrimp. The very best results are obtained when you can nearly match the algae growth rate to the growth rate of the brine shrimp.
If the brine shrimp get ahead of the algae then the water will loose its green tint, and the brine shrimp will quickly starve to death. If the algae get ahead of the brine shrimp it is really not a problem, but the brine shrimp will likely grow a bit slower in the very dense culture, and may not reach the yield that would otherwise be possible. The amount of eggs you add to the medium will affect the size of the brine shrimp at harvest. If you start with a lot of shrimp you may exhaust the supply of algae before the shrimp get very large. If you start with a small amount of shrimp, they may grow to maturity and beyond (i.e. spawn and give birth to young) before the algae is fully consumed.
You should also keep in mind that the risk of contamination by blue-green algae or bacteria increases the longer the culture is growing. After the water in the cultures becomes slightly brown and possibly very opaque, you need to harvest the shrimp and toss all the water they were in.
[Revision:You can get a very good idea of when you are getting close to needing to harvest the shrimp by removing the bubbler and taking a close look at the bottle. If you see more than a few dead large shrimp swirling around in the bottle, then your culture is very close to crashing. Within 12 hours all shrimp may be dead (what a waste). Harvest before it is too late. ]
If you don't need the shrimp right away, you can transfer them to a new culture of micro algae, or just to new, clean salt water, and keep them growing. If your shrimp get ahead of the algae and you think the water is still clean enough, you can add food for the shrimp to extend the 'life' of the water a bit (you will also of course need to feed any shrimp you transfer to fresh salt water).
Part 2
Other Methods
The second method is essentially the same as the first, except you hatch the brine shrimp in a separate container, separate them from the spent eggs using the usual methods (settling or light), and them rinse them a bit in clean water (either salt or fresh) before adding them to the algae cultures. Some eggs have a lot of various contaminants on them, so this method will keep your initial inoculum cleaner.
The third method is to wait until the algae cultures become very dense and then add your shrimp. The advantage is that there is no chance the shrimp will get ahead of the algae, the disadvantage is that the shrimp may grow slower in extremely dense algae cultures.
With any of these methods you can add inert foods to extend the 'life' of the culture water. You can use dried yeast (best to inactivate or kill first by heating), egg yolk, or flour (I've heard soy flour is good). You can also use RotiRich from Florida Aqua Farms, or Selcon, or Selco (these last two can enhance the nutritional value of the shrimp, so I've been told). Any inert food should be finely divided so that it stays in suspension and the shrimp can get it. Also, after you add inert food, you will probably need to harvest the shrimp within 1 to 2 days as the water will foul quickly with bacteria, etc.
[Revision: This is not always true. If your algae partially falls out of solution, and starts to grow on the walls of the container it will for a miniature biofilter for the bottle. The water will typically become clear in this case, both due to the brine shrimp uptake of suspended particles, and due to the algae (there is likely more than just starting algae species growing now)taking up contaminants. Remember, however that you will always get a larger harvest per volume of water used if you can keep the algae in suspension from the beginning of the culture.
Some other notes about the procedures above
1. You can grow the shrimp on inert foods alone, but it is very difficult to get the shrimp to survive the first 5 days or so of growth. I don't recommend this method because you will likely use a lot of water and end up with only a few shrimp. Also, shrimp grown on micro algae are supposed to be more nutritious. They certainly swim a lot faster, and look a lot healthier than inert grown shrimp.
2. Temperature: Best temperature is probably about 80 F. You will get reasonable growth from 70-85 F. I've read that San Francisco shrimp tend to have a lower optimum temperature than shrimp from the Great Salt Lake.
3. How Long? Under near optimum conditions shrimp grow to full size and begin to mate and develop egg sacks within 2 weeks.
4. My algae doesn't stay in solution!! Don't worry, be happy. Just try to shake your flasks up by hand once a day or so. Cap them with the original caps and shake the hell out of them for 20 seconds or so. As the culture grows it may be more likely to stay in solution. Low micro algae inoculation densities can lead to this problem. In some cases the brine shrimp themselves will help with this problem. Larger, older brine shrimp can actually scrape the algae off the bottom of the bottle.
[Revision: this can be a recurring problem. One thing to keep in mind, if you can get a bottle that for whatever reason the algae is staying in solution, use this bottle to inoculate other bottles, and do not try to expand too fast. You may have to expand only 1 to 1, or 1 to 2 occasionally. ]
5. Why not use airstones to get more air (i.e. CO2) into the culture to make the algae grow faster? Not worth the trouble. Air coming through airstones will not agitate the water in the bottle as well. I believe one of the limiting factors is keeping the algae in solution. If the number of air pumps is not a problem, you might try using both an airstone and a regular bubbler (I've not done this experiment yet). I've also heard that fine bubbles may damage both micro algae cells and brine shrimp, but I've not experienced this first hand. Finally, air stones will plug, and are really a pain to try to clean.
6. My bottles are all set and the bubbler is going like mad, but the shrimp seem to really be getting tossed around a lot. Won't they die? NO. The shrimp do extremely well under these conditions. As long the bubbler is not bubbling so vigorously that the water is splashing out the the bottle, you are okay. Amazing as it may seem, the shrimp can grow to full size and latch onto each other and mate under these conditions. Don't ask me how, consult the Kama Sutra of Artemia salina.
[Revision: the previous statement is not completely accurate. When the shrimp are small you can pound them around all you want (the more circulation the better to keep the algae in suspension and growing quickly), but as the shrimp grow they slowly become more susseptible to the shear forces of the water bubbling so hard...So...slow it down a bit. Usually this is not a problem since by this time the culture is doing well (thealgae is staying in solution) so really vigerous bubbling is not as important. ]
7. I was growing up some shrimp that reached near full size. The water was starting to go bad, but I didn't need the shrimp just yet so I transferred them to a new algae culture, and now they seem to be dying. What's wrong? Good question, I'm not sure. It may be that larger, older shrimp do not adapt as well to a sudden change in their water conditions (i.e. pH etc). It may be helpful to transfer some of the water from the old bottle to the new bottle. Keep in mind that the younger the shrimp the more adaptable the shrimp.
8. If your shrimp start to grow slowly and/or your water seems unusually opaque you may have some sort of contaminant in your cultures (blue-green algae, or possibly a high level of bacteria). You may need to occasionally clean up all the equipment that contacts your cultures, and then sterilize it as best as possible with heat, or chemicals (bleach, sodium hydroxide, or hydrochloric acid).
[Revision: I now have tubing connectors in line just before the connections to my rigid airline tubing bubblers. On a regular basis I remove the bubblers and put them in a two liter bottle filled to the top with a dilute bleach solution. I leave them there for several days, and then rinse off quite a few at once with tap water. I now try to use a clean bubbler whenever I am starting a new culture. ]
9. What about the photoperiod? It doesn't seem to matter much. Brine shrimp will grow to maturity and reproduce even if the lights are NEVER turned off. I currently leave the lights on continuously to increase the rate of algae growth and therefore brine shrimp growth. Keep in mind that the brine shrimp can grow very quickly in optimal conditions, usually the amount of algae you can grow will be the rate limiter.
10. Can I store live brine shrimp? YES. Previously I'd had problems storing live brine shrimp that I would bring home from the pet shop. I'd put it in the frig., but the die off would be fast, and after 3 days they were nearly all dead. Fortunately, I can tell you that the brine shrimp you are growing are much healthier than the ones you get from the LFS. If you want to store brine shrimp it is best to use the healthiest shrimp you have (i.e. from a culture that clearly has lots of microalgae still available and therefore has well fed brine shrimp, and from a culture that is not too far along, i.e. not too opaque yet). You can just pour the water into a small container and put it in the frig, or you can concentrate the shrimp first with a net and then put them in a small container with some good microalgae in the water. You can also add Selcon to the water. Since the shrimp will metabolize so slowly at frig. temps, the small amount of space, and food will not be a problem. I've stored shrimp this way for more than a week and had them still look quite healthy when I fed them to my seahorses.
11. This method of production seems to work okay, but now I want to scale up by about a factor of 10. I'd like to use the extra aquariums that I have. Will it work? Maybe. I've tried many times to grow the brine shrimp in larger volumes of water, but I've never been able to raise a large number of shrimp. I have always ended up wasting large amounts of water and having to clean up big messes in the aquariums. Obviously some people are successful at this large scale, but they may have access to large volumes of sea water, or use sophisticated filtration systems, etc. I don't know their tricks. I've often thought about putting a large children's wading pool outside, inoculating it with algae, then a few days later with brine shrimp. The problem as I see it is to keep the algae in suspension. At first you could use a power head, but when you add the brine shrimp you will end up with a frothy brine shrimp milkshake as the brine shrimp are shredded by the power head. Perhaps some sort of gentle agitation like a paddle wheel would work....hmmm.
I hope this how to manual will help you grow the amount of brine shrimp you want.
Happy culturing.
Gregory W. Hiller
Revision:
I've done a few more experiments with larger scale culture, and somewhat less intensive methods of late. What I've found is that if you are doing the intensive, small scale work I wrote about above then strong aeration can only help. However, if you use inert foods and/or larger tanks you may be better off NOT trying as hard to keep things in suspension. I believe the problem is one of bacterial growth. If the bacterial growth is very fast as might be the case with a culture with large amounts of inert material floating about, then the bacterial may harm the brine shrimp either just by their presence, or by the fact that they are consuming all of the available oxygen. So paradoxically, you may have higher oxygen levels if you DON'T try so hard to keep things in suspension. The brine shrimp can also eat the bacteria and will clear the water on their own to a certain extent. In some cases the easiest (but less intensive) method to culture the brine shrimp is just to toss them into a big tub and throw some algae, or inert foods in occasionally. Then just net a few out occasionally and rinse them off. This also works well outside in the sun during the warmer weather. You may even end up with some rotifers also growing in the water.
Reproducible method for brine shrimp grow out - written by Greg Hiller 3-97, revised 6-98
I have used this method over the past two years with very good results. The consumables usage is quite low, and the labor involved is also low, but a small amount of labor is required each day for the best results/highest yields.
What you need
1. As many clear two liter coke bottles as you can get your hands on. Remove the labels. Rinse with tap water. After use it is hard to clean these, but possible if you have to. I just use tap water, hand dishwashing soap, and a brush that can reach to the bottom of the bottle. You can of course use glass bottles, etc. if you want.
2. A pure culture of Nannochloris algae (the disks are the easiest to use,~$7. Revision: I actually think a liquid culture is easier to use now. The nice thing about a disk is that in some cases you may be able to save part of the disk for use in the future (months later) in case your algae culture becomes contaminated. I'm also not really sure which algae strain is the best to use. Most public aquaria use Isochrysis galbana, which I've been told from Carolina Biological is prone to crashing. Also commonly used is Monochrysis, and Carolina tells me this is easier to grow. Although I only grew this one for a short period of time once, it did seem to grow very quickly.
3. Micro Algae Grow
4. Silicone antifoam
5. Brine shrimp eggs (for you purists, Artemia salina cysts), I recommend OSI collected from the Great Salt Lake. I have found these to hatch many timesgreater amounts of shrimp per volume of eggs. The eggs also disperse better in the water, and seem to be much cleaner based on the cloudiness of the water after hatching. I've heard that eggs from collected from the San Francisco Bay are smaller in size at hatching, but if you're interested in growing them then this is not really a concern.
6. A strong air pump (this is most likely going to be the limiting factor for the amount of brine shrimp you can grow)
7. A pair of 40 watt cool-white fluorescent lights (I bought one for $10, the longer the bulbs, the more shrimp you can grow)
8. Flexible airline tubing
9. Rigid airline tubing
10 . Synthetic salthingyer mix
Items 2 through 4 above are available from Florida Aqua Farms (352-567-8540). You can also purchase the algae culture from Carolina Biological, and their cultures tend to be a bit more predictable (1-800-334-5551). To save money on salt water mix the shrimp and algae can be grown in water with a specific gravity of about 1.018. It is very important to maintain at least one micro algae culture under aseptic (sterile) conditions. This can be done by sterilizing the water used to culture the algae. The easiest way at home is to use the microwave. A one liter glass bottle of water can be sterilized in about 7-8 minutes in most microwaves.
Alternatively you could use membrane filtration for sterilization. If you use heat for sterilization, it is best to add the salt mix, but not the micro algae grow (fertilizer)before the sterilization. After the water has cooled, carefully (trying to keep things clean) add the micro algae grow (i.e. fertilizer) to the water in the concentration recommended on the bottle. I have found that 0.6 ml or about 1/8th teasthingy works well for 1 liter of water.
Follow the instructions on the micro algae pure culture that you received to get the algae into suspension. If it is a algae disk place an 1/8th inch layer of water over the surface of the disk and leave in a moderately lit area for a day or so. Rub the algae off the surface with a sterile Q-tip. Pour the suspension into asterile bottle with your fertilized, sterile salt water, place the bottle in front of your fluorescent lights (side as opposed to above lighting is best), and put your bubbler in place. The bubbler runs air from your air pump to the bottle. You can cut the rigid plastic tubing a few inches longer than the depth of the bottle, then using a cigarette lighter heat the tubing and put a 90 degree bend in the tubing so that the bubbling end of the tubing is just slightly off the bottom of the bottle when it is in place. For the bottle with your pure algae culture, try to use a tight fitting cap with a hole in it for the bubbler. You may need to loosen it a bit for the air to escape. The air that runs into your pure culture should probably run through a sterilizing filter before going into your culture.
You can prepare a homemade sterilizing filter by stuffing some filter floss or glass wool into a large diameter piece of tubing. Remember not to add too much floss, or you will not get enough air into your culture. You want the air to really turn the water over in your culture so that the algae stays in solution. The algae culture may take some time to adapt to your water conditions, etc., so at first it may sometimes fall out of solution a bit.
When the algae culture has grown to a point at which you can no longer see through the bottle, you can expand the culture. First prepare a new bottle that will hold your sterile, stock culture. You can typically expand your culture about 3 fold without any difficulty. If you try to expand further than about 7 fold you may risk having the culture be more difficult to keep in suspension, or suffer a very long lag time before the culture comes back up to a high density. After you start the new algae culture, you can use the rest of the algae for your brine shrimp growth.
I don't try to keep the bottles that I use for the brine shrimp growth under sterile conditions, although I do try to keep things clean as much as possible. As long as you carefully maintain a pure culture of the algae, and go back to it on a regular basis to start your expanded cultures for brine shrimp growth, you should not have a problem with contamination. The principal contamination problem that I have incurred is with what I believe was a blue-green algae. Over the course of a few weeks my algae culture seemed to grow faster than I had previously recalled, and grow to a very opaque consistency. Also, the brine shrimp I put into the algae seemed to grow extremely slowly, so much so that there was no point in continuing to attempt to culture them.
[Revision: I have had other types of contaminations: sometimes the algae will form stringy clumps, or diatoms will grow on the sides of the two liter bottles. Fortunately, these types of contamination do not kill or significantly slow the growth of the brine shrimp. The only limiting factor in this case is the problem that the brine shrimp can only eat small particles, and the smaller the brine shrimp the smaller the particles it can consume. FWIW recently I've heard that these stringy clumps are formed by a type of fungi that can grow in the cultures. I've also been told that it can be effectively removed by filtering the culture through a net, the micro algae that you want will flow through]
For the brine shrimp growth you can use several different methods depending upon how much time you want to spend, and how much brine shrimp you want to recover. The first method is to hatch the brine shrimp directly in the bottles you will use for grow out. This means you expand your left over algae culture (after starting a new culture) to a 2 liter bottle, and put a very small pinch of brine shrimp eggs in the bottle. You also need to fertilize the added salt water, and then add a drop, or dab of the silicone antifoam.
[Revision: Keep the antifoam in a sealed container that is opaque to keep out light, things will grow in it otherwise. ]
If you do not use the antifoam, when your cultures start to grow and you get some protein production, a large number of the shrimp will die as they are trapped in the foam layer that forms (this 'aquarium' is a bit small to use a protein skimmer!). Fill your 2 liter bottles only until you reach the area at the top of the bottle where the neck begins to narrow. The trick with this method of culture (as with all the rest I will mention) is that you need to keep the algae culture growing a bit faster than the brine shrimp. The very best results are obtained when you can nearly match the algae growth rate to the growth rate of the brine shrimp.
If the brine shrimp get ahead of the algae then the water will loose its green tint, and the brine shrimp will quickly starve to death. If the algae get ahead of the brine shrimp it is really not a problem, but the brine shrimp will likely grow a bit slower in the very dense culture, and may not reach the yield that would otherwise be possible. The amount of eggs you add to the medium will affect the size of the brine shrimp at harvest. If you start with a lot of shrimp you may exhaust the supply of algae before the shrimp get very large. If you start with a small amount of shrimp, they may grow to maturity and beyond (i.e. spawn and give birth to young) before the algae is fully consumed.
You should also keep in mind that the risk of contamination by blue-green algae or bacteria increases the longer the culture is growing. After the water in the cultures becomes slightly brown and possibly very opaque, you need to harvest the shrimp and toss all the water they were in.
[Revision:You can get a very good idea of when you are getting close to needing to harvest the shrimp by removing the bubbler and taking a close look at the bottle. If you see more than a few dead large shrimp swirling around in the bottle, then your culture is very close to crashing. Within 12 hours all shrimp may be dead (what a waste). Harvest before it is too late. ]
If you don't need the shrimp right away, you can transfer them to a new culture of micro algae, or just to new, clean salt water, and keep them growing. If your shrimp get ahead of the algae and you think the water is still clean enough, you can add food for the shrimp to extend the 'life' of the water a bit (you will also of course need to feed any shrimp you transfer to fresh salt water).
Part 2
Other Methods
The second method is essentially the same as the first, except you hatch the brine shrimp in a separate container, separate them from the spent eggs using the usual methods (settling or light), and them rinse them a bit in clean water (either salt or fresh) before adding them to the algae cultures. Some eggs have a lot of various contaminants on them, so this method will keep your initial inoculum cleaner.
The third method is to wait until the algae cultures become very dense and then add your shrimp. The advantage is that there is no chance the shrimp will get ahead of the algae, the disadvantage is that the shrimp may grow slower in extremely dense algae cultures.
With any of these methods you can add inert foods to extend the 'life' of the culture water. You can use dried yeast (best to inactivate or kill first by heating), egg yolk, or flour (I've heard soy flour is good). You can also use RotiRich from Florida Aqua Farms, or Selcon, or Selco (these last two can enhance the nutritional value of the shrimp, so I've been told). Any inert food should be finely divided so that it stays in suspension and the shrimp can get it. Also, after you add inert food, you will probably need to harvest the shrimp within 1 to 2 days as the water will foul quickly with bacteria, etc.
[Revision: This is not always true. If your algae partially falls out of solution, and starts to grow on the walls of the container it will for a miniature biofilter for the bottle. The water will typically become clear in this case, both due to the brine shrimp uptake of suspended particles, and due to the algae (there is likely more than just starting algae species growing now)taking up contaminants. Remember, however that you will always get a larger harvest per volume of water used if you can keep the algae in suspension from the beginning of the culture.
Some other notes about the procedures above
1. You can grow the shrimp on inert foods alone, but it is very difficult to get the shrimp to survive the first 5 days or so of growth. I don't recommend this method because you will likely use a lot of water and end up with only a few shrimp. Also, shrimp grown on micro algae are supposed to be more nutritious. They certainly swim a lot faster, and look a lot healthier than inert grown shrimp.
2. Temperature: Best temperature is probably about 80 F. You will get reasonable growth from 70-85 F. I've read that San Francisco shrimp tend to have a lower optimum temperature than shrimp from the Great Salt Lake.
3. How Long? Under near optimum conditions shrimp grow to full size and begin to mate and develop egg sacks within 2 weeks.
4. My algae doesn't stay in solution!! Don't worry, be happy. Just try to shake your flasks up by hand once a day or so. Cap them with the original caps and shake the hell out of them for 20 seconds or so. As the culture grows it may be more likely to stay in solution. Low micro algae inoculation densities can lead to this problem. In some cases the brine shrimp themselves will help with this problem. Larger, older brine shrimp can actually scrape the algae off the bottom of the bottle.
[Revision: this can be a recurring problem. One thing to keep in mind, if you can get a bottle that for whatever reason the algae is staying in solution, use this bottle to inoculate other bottles, and do not try to expand too fast. You may have to expand only 1 to 1, or 1 to 2 occasionally. ]
5. Why not use airstones to get more air (i.e. CO2) into the culture to make the algae grow faster? Not worth the trouble. Air coming through airstones will not agitate the water in the bottle as well. I believe one of the limiting factors is keeping the algae in solution. If the number of air pumps is not a problem, you might try using both an airstone and a regular bubbler (I've not done this experiment yet). I've also heard that fine bubbles may damage both micro algae cells and brine shrimp, but I've not experienced this first hand. Finally, air stones will plug, and are really a pain to try to clean.
6. My bottles are all set and the bubbler is going like mad, but the shrimp seem to really be getting tossed around a lot. Won't they die? NO. The shrimp do extremely well under these conditions. As long the bubbler is not bubbling so vigorously that the water is splashing out the the bottle, you are okay. Amazing as it may seem, the shrimp can grow to full size and latch onto each other and mate under these conditions. Don't ask me how, consult the Kama Sutra of Artemia salina.
[Revision: the previous statement is not completely accurate. When the shrimp are small you can pound them around all you want (the more circulation the better to keep the algae in suspension and growing quickly), but as the shrimp grow they slowly become more susseptible to the shear forces of the water bubbling so hard...So...slow it down a bit. Usually this is not a problem since by this time the culture is doing well (thealgae is staying in solution) so really vigerous bubbling is not as important. ]
7. I was growing up some shrimp that reached near full size. The water was starting to go bad, but I didn't need the shrimp just yet so I transferred them to a new algae culture, and now they seem to be dying. What's wrong? Good question, I'm not sure. It may be that larger, older shrimp do not adapt as well to a sudden change in their water conditions (i.e. pH etc). It may be helpful to transfer some of the water from the old bottle to the new bottle. Keep in mind that the younger the shrimp the more adaptable the shrimp.
8. If your shrimp start to grow slowly and/or your water seems unusually opaque you may have some sort of contaminant in your cultures (blue-green algae, or possibly a high level of bacteria). You may need to occasionally clean up all the equipment that contacts your cultures, and then sterilize it as best as possible with heat, or chemicals (bleach, sodium hydroxide, or hydrochloric acid).
[Revision: I now have tubing connectors in line just before the connections to my rigid airline tubing bubblers. On a regular basis I remove the bubblers and put them in a two liter bottle filled to the top with a dilute bleach solution. I leave them there for several days, and then rinse off quite a few at once with tap water. I now try to use a clean bubbler whenever I am starting a new culture. ]
9. What about the photoperiod? It doesn't seem to matter much. Brine shrimp will grow to maturity and reproduce even if the lights are NEVER turned off. I currently leave the lights on continuously to increase the rate of algae growth and therefore brine shrimp growth. Keep in mind that the brine shrimp can grow very quickly in optimal conditions, usually the amount of algae you can grow will be the rate limiter.
10. Can I store live brine shrimp? YES. Previously I'd had problems storing live brine shrimp that I would bring home from the pet shop. I'd put it in the frig., but the die off would be fast, and after 3 days they were nearly all dead. Fortunately, I can tell you that the brine shrimp you are growing are much healthier than the ones you get from the LFS. If you want to store brine shrimp it is best to use the healthiest shrimp you have (i.e. from a culture that clearly has lots of microalgae still available and therefore has well fed brine shrimp, and from a culture that is not too far along, i.e. not too opaque yet). You can just pour the water into a small container and put it in the frig, or you can concentrate the shrimp first with a net and then put them in a small container with some good microalgae in the water. You can also add Selcon to the water. Since the shrimp will metabolize so slowly at frig. temps, the small amount of space, and food will not be a problem. I've stored shrimp this way for more than a week and had them still look quite healthy when I fed them to my seahorses.
11. This method of production seems to work okay, but now I want to scale up by about a factor of 10. I'd like to use the extra aquariums that I have. Will it work? Maybe. I've tried many times to grow the brine shrimp in larger volumes of water, but I've never been able to raise a large number of shrimp. I have always ended up wasting large amounts of water and having to clean up big messes in the aquariums. Obviously some people are successful at this large scale, but they may have access to large volumes of sea water, or use sophisticated filtration systems, etc. I don't know their tricks. I've often thought about putting a large children's wading pool outside, inoculating it with algae, then a few days later with brine shrimp. The problem as I see it is to keep the algae in suspension. At first you could use a power head, but when you add the brine shrimp you will end up with a frothy brine shrimp milkshake as the brine shrimp are shredded by the power head. Perhaps some sort of gentle agitation like a paddle wheel would work....hmmm.
I hope this how to manual will help you grow the amount of brine shrimp you want.
Happy culturing.
Gregory W. Hiller
Revision:
I've done a few more experiments with larger scale culture, and somewhat less intensive methods of late. What I've found is that if you are doing the intensive, small scale work I wrote about above then strong aeration can only help. However, if you use inert foods and/or larger tanks you may be better off NOT trying as hard to keep things in suspension. I believe the problem is one of bacterial growth. If the bacterial growth is very fast as might be the case with a culture with large amounts of inert material floating about, then the bacterial may harm the brine shrimp either just by their presence, or by the fact that they are consuming all of the available oxygen. So paradoxically, you may have higher oxygen levels if you DON'T try so hard to keep things in suspension. The brine shrimp can also eat the bacteria and will clear the water on their own to a certain extent. In some cases the easiest (but less intensive) method to culture the brine shrimp is just to toss them into a big tub and throw some algae, or inert foods in occasionally. Then just net a few out occasionally and rinse them off. This also works well outside in the sun during the warmer weather. You may even end up with some rotifers also growing in the water.