Construction of an Affordable and Easy-to-Build Zebrafish Facility

The overall goal of this procedure is to build an affordable zebra fish facility that requires minimal maintenance. This is accomplished by first laying out the PVC pieces, reservoir, pump and filters and assembling them using PVC primer and glue before attaching them to the shelf. The second step is to assemble the supply lines to each of the three shelves, making sure to include a pressure relief line.

Next, the fish tank exhaust lines are constructed and connected to a main exhaust line. The final step is to prepare the system for the fish by performing a 72 hour test run and optimizing water quality. Ultimately, this facility is optimized for a variety of zebra fish research techniques at a small institution.

The main advantage of this facility construction of our existing methods is that the construction is simple and takes only a few hours to complete. The system is covered, which makes water quality control much easier. Though this facility is simple to build, it can also be modified to fit your research needs by making small changes such as making your tank larger to accommodate bigger colonies or different species of fish.

We first had the idea for this method when we found existing construction plans unclear, complicated to build and open to contamination. I was a primary builder of the zebrafish module we will be looking at today. This zebrafish module is fully operational, so I will cover the basic techniques and assembly overview.

Keep in mind your assembly may vary based on the size and components used in your facility. To begin construction, lay out the 18 gallon bucket pump filters and UV lamp to determine the final installation location and size. Once the parts are laid out, cut the three fourths inch PVC pipe to the length needed for assembly.

Given that the facility is already built, we will show you how to cut the PVC fittings. Keep the pipes and fittings in their position and sand the cut edges of the pipe until they are smooth to the touch and there is no debris. Next, obtain the purchased PVC primer and use the included swab to prime the orifice of each fitting.

That will be cemented together. Apply PVC cement to the outside of the pipe to join the pipe and fitting together. When applying PVC primer or PVC cement, make sure to perform the procedure in a well ventilated area.

Then place the 18 gallon bucket to one side of the shelf and install a bulk head connector in the side of the reservoir tank. Attach a three fourths inch PVC pipe to the bulk head and attach the other end of the three fourths inch pipe to the pump inlet. Using a three fourths inch threaded make fitting.

Next, connect the filters to the pump using two 90 degree elbows and three fourths inch PVC pipe. Use one paper filter followed by one activated carbon filter to remove debris and reduce chlorine and chemical levels in the water. Connect the UV lamp to the filters according to the manufacturer's instructions.

Each installation will depend on the type of pump purchased. It is important to note that each pump will be different. Then attach the pump and filter system to the top of the first shelf of the rack.

Using zip ties vary the order of the assembly and attachment according to the size and layout of the shelf or rack chosen. Once the pump and filter system are assembled and attach to the shelf, add one bio ball per gallon of water to the water reservoir measure and cut a piece of three fourths inch PVC pipe and connect the T to the three fourths inch PVC exhaust coming from the UV filter. To start the main supply line, add an on off valve to the bottom of the PVCT to serve as a drain to maintain system flushing.

Make sure to measure the PVC pipe according to the dimensions of the specific shelf in use. After adding the valve measure and cut the three fourths inch PVC pipe to provide feed pipes for each of the shelves on the rack. To start the main supply line, repeat for each of the shelves in the system.

Then install three fourths inch PV cts for the horizontal lines that will enter each shelf on the rack and add a three fourths inch on off valve for each shelf supply line to allow the isolation of each shelf and a modular design of the system repeat for each of the shelves in the system. Next, attach a three fourths inch PVC pipe on the top shelf as a pressure reducing line. End the pressure reducing line in the main exhaust line with a three fourths inch on off valve.

After the main supply line has been set up, prime and cement the PVC pipe together. Run the vertical length of the main exhaust line to the other side of the rack and use a two inch PVC pipe to receive the pressure reducing line at the top and the drain lines from the fish tanks from each shelf and this line in the stock tank to expel the recycled water over a sponge filter. After marking where each fish tank will empty into the horizontal drain line, drill holes in your exhaust line for each individual tank exhaust after holes have been cut into the line, apply two inch PVC caps to the horizontal drain lines.

To finish the main exhaust line, prime and cement the pieces using connectors, nipples, or elbows with the drill. Cut a one and one eight inch hole one inch below the upper edge of the back of the fish tank. Then slide a threaded three fourths inch PVC male adapter with a number 18 O ring attached through the hole.

Next, attach and tighten a 90 degree three fourths inch threaded PVC elbow to the male adapter to create the fish tank exhaust pipe. Add a piece of fine mesh before adding another number 18 O ring to secure the mesh to prevent the zebra fish from passing through the exhaust system. To avoid potential leaks, glue the external part of the hole where the elbow is placed.

Follow manufacturer's instructions for drying time. Then place the exhaust pipe side of the fish tank into the open space of the draining line behind the tanks from the stop on off valve measure and cut a piece of PVC that is half of the length of the shelf and place a 90 degree elbow on the end with an adapter to attach the supply. Then attach a three fourths inch to one half inch male adapter to the supply line measure and cut the vinyl tubing for each individual tank and attach to the irrigation diverter.

Next thread, the one half inch male end wrapped in Teflon tape onto the nine way irrigation. Diverter a affix the irrigation supply lines to the top shelves using zip ties. Once the beginning of each line is secure, add an irrigation on off stop valve to each tube.

Then secure the end of each line with binder clips and more zip ties. Beginning with the horizontal supply stop valve. Slowly open the valve and attenuate the flow to the desired level.

Measure out three times the amount of water contained within the system and allow it to stand at room temperature for 48 hours. It is important to note that water should not have been in the system until this point. After 48 hours, add the conditioned water to the system and let it circulate for three days vigorously change the water in the system throughout the initial three days of circulation.

In order to rid the system of contaminants that reside within the pipes. Add seven starter zebra fish per 1.8 liters and allow the nitrogen cycle to establish itself. Normalize the pH to a consistent seven.

Feed the zebra fish twice daily during weekdays and once daily during weekends. Finally, monitor the balance of nitrate nitrite and ammonium daily during the first week after adding the fish and twice weekly. Once these three photos are examples of individual components of the system, the two filter and pump system is located on the bottom rack.

The pump is connected to two mechanical filters and a UV filter to eliminate contaminants from the water. The horizontal supply line ends in a nine port irrigation manifold, which supplies fresh water and oxygen to each tank. The supply line has an on off valve that can be used to stop water to each shelf for cleaning and quarantine.

The fish tank exhaust line and pressure relief line empty into the main exhaust line to reenter circulation. The completed system is comprised of a pump and filter system, a main vertical supply line, a horizontal supply line for each shelf, a tank exhaust line, a pressure relief line, and a main exhaust line for a contained circulation system. This facility can be constructed in less than four hours if all the preparations are made.

Accordingly, While attempting this procedure, it's important to remember to be precise in your measurements and layout. After watching this video, you should have a good understanding of how to build a zebrafish facility, which can be used to perform your chosen experiments.