This protocol describes the regular care and maintenance of a zebra fish laboratory. Due to the spare of interest in zebra fish research, the need to establish and maintain a zebra fish housing facility is also increasing. Although literature is available for the maintenance of a zebra fish laboratory, a concise video protocol is lacking.
This video explains system maintenance, feeding, breeding, and raising of zebrafish larvae. Hi, I am ABH from School of Medical Sciences at University. Hi, I'm Manchi from Thee Institution.
Today we are going to show you regular care and maintenance of a zebrafish laboratory. Zebrafish are becoming popular in genetics, pharmacological, and behavioral research. Due to their high F ease of maintenance and rapid development, They are highly amenable to genetic modulation and suitable for high stroke drug screen.
This part code provide ideas of zebrafish husbandry to those fish users and the researchers who are considering of establishing new fish facility. So let's get started. Several companies provide fish systems, but we use systems from aquatic habitats.
In our laboratory, zebra fish are kept at 26 to 28.5 degrees Celsius and a 14 to 10 light to dark cycle. Zebra fish are kept in a circulating system, which continuously filters and aerates the system water. To maintain the water quality required for a healthy aquatic environment, the system utilizes a set of filters of different types within our system.
Water from the tanks passes through a filter pad, canister filter, biological filter, active carbon filter, and UV filter before being circulated back into the tank. Dechlorinated or aged water is used in the zebrafish system. Water can be dechlorinated by aging for at least 48 hours.
Check system water pH daily and maintain between 6.8 and 7.5. Using sodium bicarbonate, fish tanks should be cleaned regularly. To clean a fish tank, close the water flow to this tank, drain excess water by tilting the tank backwards and remove the tank carefully from the system.
Set up a clean tank and fill it with system water. Carefully transfer the fish into this tank with a fish net. Close the lid and replace the tank name tag carefully.
Place the tank in the system. Place the tube back into the tank and switch on the water supply. Decontaminate the fish net and tank with 70%ethanol.
Rinse in water and let it dry. Before reusing. Filters of the circulating system have to be checked and changed regularly to ensure their proper function.
The 120 micron filter pad is checked and repositioned towards the water flow every day before being replaced. The canister filter should be changed weekly. To change the canister filter, remove it with a wrench or hands.
Place a plastic sheet or towel below to prevent water spillage. Insert a new canister, filter and fit the unit back into the system. Carefully tighten with hands and with a wrench.
If necessary, the carbon filter should be changed fortnightly. To change the carbon filter, remove it carefully. With a wrench in hands, discard the activated carbon from the filter and replace it with new activated carbon.
Refit the carbon holder and place it back into the filter unit. Fit the filter unit back into the system. Switch on the system and check if the water is flowing to the filter.
The biological filter should be cleaned every six months. To change the biological filter, remove the filter from the system. Release the pressure from the filter by pressing the pressure release button.
Unscrew the filter and remove the lid. Empty the contents into a sift container to separate the Cy Oex from the water. If the Cy Oex is very dirty, replace it.
Fill the filter with the new cy ax. Fill the unit to system water. Close the lid of the filter and refit the unit and switch on.
UV filters are used to control biological contaminants and should be replaced every nine to 10 months. Zebra fish can be fed with dry food or live foods such as brine shrimps, brine shrimp, eggs are available from local pet shops and can be hatched in the laboratory by following the simple steps described here. Dissolve red sea salt in system water to make a solution of 30 to 35 grams per liter.
Fill up the brine shrimp patcher with the salt water and add the shrimp eggs at a concentration of 1.2 tablespoons per liter. Aerate the hatcher vigorously with an air pump and leave the brine shrimp eggs to fertilize for about 48 hours. Remove the air pipe and let the culture settle for four to five minutes.
The fertilized brine shrimp gather at the bottom of the hatcher using the tap at the bottom of the hatcher, discard the first flow, which contains unfertilized eggs. Then collect the fertilized brine shrimp. Separate the brine shrimp from the salty water.
Using a brine shrimp collection net. Rinse the brine shrimp into a container Using system water, the brine shrimp are present in high concentration at the bottom of the container and are now ready for zebra fish feeding. Feed the brine shrimp to the fish using a pipee or a dropper.
The amount of food dispensed depends on the population size of individual tanks. After injecting food into the water hungry fish swim to catch the brine shrimp, which is important for their regular exercise. Dry feeding can be performed either using an aquatic habitats, fish food dispenser, or using a simple spoon.
Zebra fish initiate breeding at the onset of light. Eggs can be obtained through inta or pairwise breeding. While inta breeding is more labor efficient and is used in regular embryo collection, pairwise breathing is preferred when genes or mutations are to be screened for in tank breeding, assemble the in tank breeder and drop slowly into a tank.
After the onset of light, leave the breeding set up for around 15 minutes in the tank to let the fish mate before removing the breeder from the tank and collecting the eggs. Pear wise breeding is set up late in the afternoon by assembling the breeding tank and filling it with system water. Transfer one female and one male into opposite sides of the tank.
Females can be distinguished from males because of their bigger belly. Remove the divider next morning. Shortly after the onset of light, leave the fish to mate underst disturbed for 20 minutes or until sufficient numbers of embryos are laid at the bottom of the tank.
After breeding, return the fish to their tanks. Collect the eggs with a strainer. Wash the embryos thoroughly.
With system water. Transfer the embryos to a petri dish by rinsing the strainer with embryo. Medium embryos can be observed under a microscope.
Unfertilized embryos should be separated from fertilized embryos using a needle and pipet. After the fertilized eggs are separated from the unfertilized eggs, they're kept in an incubator at around 28.5 degrees Celsius for 72 hours until the larva are hatched. Now the larvae are outta the CORs and swimming freely.
Larvae need to be fed from five days post fertilization and kept in stagnant water for a few days before transferring them to the system. Transfer the larvae gently into a tank containing a small size baffle of around 300 to 400 microns. After 14 days, larvae tanks can be shelved into the system and supplied with a small stream of cycling water.
Different sized baffles should be used depending on the size of the larvae and the adult fish. It usually takes three months for the embryos to develop into sexually mature adults. Thanks for watching the menu, and good luck for your experiments.
