IAP Trocho Feed Thawing Procedures

IAP TrochoFeed Thawing Procedures

Important safety precautions

Always wear protective eye wear and gloves when handling cryopreserved Trochofeed. Cryogenic sample holders, including vials, tubes and straws, can shatter. Cryogenic storage vessels, including dewars and refrigerators, are supplied with manufacturer 's safety guidelines which should always be followed. Never seal a cryogenic container as this will cause pressure to build up from evaporating nitrogen.

1. Selecting material

Select a straw of cryopreserved trochophores and place it immediately into the plastic thawing bag provided. The bag is for convenience in thawing, and for your protection in the unlikely event of straw breakage. Holding the outside of the bag, immediately snap the straw into 2-3 inch segments. If you're not using the whole straw, replace the unused portion in the liquid nitrogen storage container immediately. DO NOT ALLOW UNUSED PORTION TO THAW.

2. Thawing

Add warm seawater (30° C) to the bag without delay, using about 300 mI per straw. Shake gently, squeezing the TrochoFeed out with your fingers to achieve a rapid thaw. The straw pieces must be submerged to get the right thaw rate. Remove empty straw pieces.

3. Recovery - warm water applications

Filter the thawed trochophores over a 20µ Nitex screen, and wash with a few squirts of25° C seawater. Rinse the washed trochophores directly into the larval rearing tanks for feeding.

4. Recovery - cold water applications

Filter the thawed trochophores over a 20µ Nitex screen, then rinse them into a container of 25° C seawater for an approximately one hour recovery period. Density during the recovery period should be roughly 100 ml water per million trochophores (10,000 trochophores/ ml). After one hour, harvest the trochophores on the 20µ screen as before, then rinse them directly into the rearing tanks for feeding.

5. Counts and feeding rate

Trochophores are counted by Coulter counter and the total per straw is stated on your invoice. You can thaw less than an entire straw, e.g. if one straw contains 30 million trochophores, thawing half would give you 15 million trochophores.

A suggested feeding rate is 10-20 trochophores per ml of tank water per feeding, depending on the species.

6. Thaw too many?

Unused trochophores will keep overnight in a refrigerator (4-5°C).

7. Nitrogen Level

It is best to transfer the TrochoFeed from the Dry shipper into a "Liquid Nitrogen Refrigerator" as soon as possible. Keep it "topped-up" with liquid nitrogen (once a week is sufficient).

IAP Trocho Feed Information

IAP Trocho Feed Frequently Asked Questions

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