Frequently Asked Questions

• How often do you need to change the consumables and clean the system components?
• What water level should be maintained in the sump tank?
• Do you need to pre-filter the water supply before it enters the system?
• What is a Fluidized Bed Biofilter (FBB)?
• How do you acclimate the FBB?
• Do you need to clean or change the FBB?
• How do you know if the FBB is functioning properly?
• What is the capacity for zebrafish per liter? For Xenopus laevis?
• There is no water flow to the upper-shelf tanks. How do you adjust the water flow to the tanks (perform a flow test)?
• How much water should be exchanged in the system per day?
• How do you clean Aquaneering tanks? Can you use a cage wash?
• What is the approximate volume of each aquatic housing system?
• Why is algae growing in the fish tanks?
• What are the standard water quality parameters for zebrafish?
• What are the standard water quality parameters for Xenopus laevis and Xenopus tropicalis?
• Why has the rack pump shut down, and there is no water flow to the tanks?
• Why have ammonia levels increased in the system?
• I have low pH / high ammonia levels; how do I get my water quality back to normal?

How often do you need to change the consumables and clean the system components?

To request a customized maintenance placard for your system, please contact Aquaneering.

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What water level should be maintained in the sump tank?
The overflow (white PVC pipe) should be adjusted by gently twisting to maintain the water at a height of 12-14 inches. Make sure the float valve is set at a lower height than the overflow.

Do you need to pre-filter the water supply before it enters the system?
It is not necessary to pre-filter the water if the system is connected to a building reverse osmosis (RO) line. However, if you are using well or city water, you may need a carbon filter to remove chlorine in the water. Test your water supply for chlorine. If you find detectable levels of chlorine in the water, you will need a carbon filter. For assistance or other solutions, contact Aquaneering.
The water may also need to be particulate filtered. Many zebrafish researchers use an RO Water Maker, which has both carbon and particulate filters.

What is a Fluidized Bed Biofilter (FBB)?
A Fluidized Bed Biofilter is a biological filter in which the upwelling of a fine media in the water allows the growth of beneficial bacteria that aid in the nitrification process: the conversion of ammonia to nitrite to nitrate to free nitrogen. Two different bacteria are required in the nitrification process, Nitrosomonas, which convert ammonia to nitrite, and Nitrobacter, which convert nitrite to nitrate.
See our Fluidized Bed Biofilter section for more information and product specifications.

How do you acclimate the FBB?
Like all biofilters, the FBB requires an initial acclimation period for the bacteria to grow and convert the ammonia to nitrite to non-toxic nitrate.
The quickest method to establish a bacterial colony in an FBB is to siphon several buckets of media from an operating FBB and add it to the new FBB.

On a new system, it will take 6 to 8 weeks to establish the bacteria in the FBB.

1. Fill the system with water and allow it to run for a day. The water temperature should be between 26˚ to 28˚ C.
2. Add approximately 10% of the system’s capacity of animals. On a 5-Shelf Stand Alone System, 10% would equal about 100 adult zebrafish. Do not feed the animals for the first 1 - 2 days to slowly allow the ammonia to build up in the system.
3. After 1 - 2 days, begin feeding the animals a reduced diet, once per day. Feed ONLY what they will eat in a minute or less. Measure the ammonia levels daily.
4. In 3 - 4 weeks, the measured levels of ammonia will start to decline. At this time, add another 100 zebrafish to the system.
5. If ammonia levels begin to climb above 1.5-2 ppm:
   a. Perform a water exchange.  A 50% water change out will reduce the ammonia level by 50%.
   b. Stop feeding the animals for 2-3 days, and then resume feeding at a reduced rate. Less food equals less waste produced, which will lower the ammonia levels and allow the bacteria to multiply to meet the demand.
   c. Stop adding animals to the system, and let the system stabilize.
   d. Make sure the filter pads are being changed 2 to 3 hours after feeding.
6. The ammonia levels will start to drop to undetectable levels in approximately 6 - 8 weeks. Until the ammonia levels drop below 0.5 ppm, only feed the animals once per day.
7. Once the ammonia level is undetectable, the animals can now be fed 2-3 times per day. Only feed what the animals will eat in 1-2 minutes.
8. Make sure to maintain the pH above 7. If the pH is below 7, add sodium bicarbonate in increments of 2 grams per day (on a Stand Alone System).
9. The acclimation process is finished when the ammonia and nitrite levels are undetectable, and the nitrate level is less than 50 ppm.

Do you need to clean or change the FBB?
The FBB is self-cleaning and never requires changing.

How do you know if the FBB is functioning properly?
Test the system water for ammonia and pH. If there is less than 0.5 ppm of ammonia, and the pH is above 6.5, the FBB is filtering properly. In addition, the media should be evenly distributed and upwelling. If the media is compacted, the pump may have shut down or a FBB probe is clogged.

What is the capacity for zebrafish per liter? For Xenopus laevis?
Zebrafish: 15 fish/liter.
Xenopus laevis: 1 frog/3 liters

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There is no water flow to the upper-shelf tanks. How do you adjust the water flow to the tanks (perform a flow test)?
In order to regulate the water pressure, you will need to perform a "10 Minute Flow Test":

1. Adjust the tank supply valve on a tank on the bottom shelf so the tank will fill completely in 10 minutes.
   NOTE:  If any tank fills faster than 10 minutes, there is not enough water pressure to fill the upper tanks.
   
2. Proceed up the shelves, adjusting the valve on each tank, until every tank fills completely in 10 minutes.

How much water should be exchanged in the system per day?
Aquaneering recommends a water exchange rate of 5-10% per day. This may vary depending on species and population load. For a Stand Alone System, you can exchange about 10% of the total tank volume per week. For larger Central Filtration Systems, you should exchange about 10% of the total tank volume per day.
NOTE: To calculate the system volume for water exchanges, use the total tank volume on the racks only.

How do you clean Aquaneering tanks? Can you use a cage wash?
Because the standard Aquaneering tanks are polycarbonate, they can be cage washed up to 250°F (121°C).
If you do not have a cage wash, a general guideline for cleaning tanks is to hand wash the tank with a sponge in hot water. Never use detergents.
Many labs use a high temperature dishwasher to wash and disinfect the tanks.  It is important to use a dishwasher that has a reverse osmosis (RO) final rinse cycle to remove traces of chlorine.

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What is the approximate volume of each aquatic housing system?
Table Top System: 220 liters/58 gallons
Five Shelf System: 320 - 490 liters/85 - 130 gallons
Six Shelf System: 340 - 510 liters/90 - 135 gallons
Sumps:
Single-sided sump = 130 liters/35 gallons
Double-sided sump = 300 liters/80 gallons
Tanks:
0.8L = 0.21 gallons
1.4L = 0.37 gallons
1.8L = 0.48 gallons
2.8L = 0.74 gallons
6.0L = 1.59 gallons
9.5L = 2.51 gallons
NOTE: To calculate the system volume for water exchanges, use the total tank volume on the racks only.

Why is algae growing in the fish tanks?
There are several factors that influence algae growth, including nitrate buildup. To reduce the amount of nitrates, you should exchange 5-10% of the system water daily. For more information and additional solutions, contact Aquaneering.

What are the standard water quality parameters for zebrafish?

Reference
Harper, Claudia, and Christian Lawrence. The Laboratory Zebrafish. Boca Raton, FL: CRC Press, 2011. Print.
You can purchase The Laboratory Zebrafish through CRC Press.

What are the standard water quality parameters for Xenopus laevis and Xenopus tropicalis?

Reference
Green, Sherril L. The Laboratory Xenopus sp. Boca Raton, FL: CRC Press, 2010. Print.
You can purchase The Laboratory Xenopus sp. through CRC Press.

Why has the rack pump shut down, and there is no water flow to the tanks?
When the water level in the sump drops too low, the float switch triggers the control box to power off, which shuts down the system. To restart the system, add water to the sump and lift the float switch.

Why have ammonia levels increased in the system?

• The bacteria have not been adequately established during initial set-up. There may be too many animals on the system.
• The pH is lower than 6.5. When the pH is too low, the bacteria stop nitrifying ammonia.
• If you have introduced antibiotics into the system, this can suppress or kill the beneficial bacteria in the Fluidized Bed Biofilter which nitrify ammonia.

I have low pH / high ammonia levels; how do I get my water quality back to normal?

1. Check and document all water quality parameters, including ammonia, nitrite, nitrate, pH, and water temperature.
2. If no bacteria is present: add 1 tablespoon of sodium bicarbonate directly into biofilter every hour. Check water quality and record data.
3. If bacteria is present: pour 1 tablespoon of sodium bicarbonate onto the filter pad under running water every hour. Check water quality and record data.
4. Stop adding sodium bicarbonate when pH is at 7.2-7.5 and let the system run overnight.
5. In the morning, check water quality and record data.
6. Test and calibrate the water exchange for 10%.
7. Monitor the water quality and water exchange over the next few days.
8. After 2-3 days of consistent water quality data without ammonia or nitrite present, add some sentinel fish to the system.
9. Continue to monitor water quality, water exchange, and sentinel fish for 1 week.

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