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Ashland NFWCO
 Midwest Region
 mtanlogo.gif (9794 bytes) Dedicated To The Tribal Aquaculture Program March 1993 - Volume 3
 http://www.fws.gov/midwest/ashland/mtanhome.html **Administrative
        Coordinator:**
        **Frank G. Stone
        (715-682-6185) Ext.****12
        U.S. Fish and Wildlife Service****Email:
        [email protected]**
      **Edited
        By:****Elizabeth W. Greiff
        (715-349-2195)
        St. Croix Tribal Nat. Res. Depart.****Email:****[email protected]**
  Topics Of Interest: The Twilight Zone Comes To
    Your Fish Hatchery Walleye Spawn Collection and Big Redd Operating Procedures Brown Trout Spawning Protocol Fish Hatchery Supplies Hatchery Tips  The Twilight Zone Comes To Your
Fish Hatchery By: MTAN I submit for your approval, the following program manager's nightmare.
Imagine you are concluding final plans to begin another spawning season. Naturally,
because of your past experience, you have learned to prepare early for this busy,
sometimes frustrating period. All the necessary steps have been attended to. You and the
hatchery staff feel your ahead of the game and ready for the season to begin. Your sitting
comfortably in your office congratulating yourself on your efficiency, when the phone
rings. The call is from your hatchery biologist who normally coordinates your entire
hatchery program. MTAN would never wish ill will on anyone, so lets just assume that
this person has informed you they have accepted another job and will not be able to direct
your spawning and fry/fingerling program.

As is often the case with many businesses, one individual may have specific
experience/training which allows only them to carry out particular tasks. What are you
going to do? The spawning season will begin soon and you just lost your most qualified
worker. The only person who really feels comfortable in knowing which valves to turn,
where everything is located and all about those new fish culture procedures you wanted to
try. How are you going to meet the demands of your program?

Elizabeth Greiff, Tribal Biologist for the St.
Croix Natural Resources Program, must have watched this show before. Because she has had
the foresight to prepare for this type of dilemma. What Ms. Greiff has done was to prepare
    a set of protocols for specific aspects of the hatchery operation. In this way,
should any one individual not be able to continue with their assigned duties, someone else
can step in. Another benefit for establishing protocols is that they help remind you of
the things that work and perhaps even those that didn't. They enable us to stay
consistent, so we are able to repeat those fish culture practices which work the best for
your specific program.

MTAN choose to reprint these protocols in the exact detail which was submitted by Ms.
Greiff. We feel even though your procedures and equipment may be different at your
facility, the important lesson we wish to illustrate, is to be as specific as possible
when writing operational protocols. When reading through these steps, we hope you will
maintain the point of view that someone else may not always have the "BEST" way
to accomplish a task. It may just be a different way. Although, by looking at alternate
ways of doing something, you may stumble onto what may be the "BEST" procedure
for your specific operation. MTAN sincerely hopes every Tribal hatchery program will
adopted a set of protocols similar to what Ms. Greiff has developed. But then again maybe
you enjoy living on the edge, you know, living in the

T w i l i g h t   Z
o n e

ST.
CROIX TRIBAL NATURAL RESOURCES DEPARTMENT WALLEYE SPAWN COLLECTION PROCEDUREBy: Elizabeth W. Greiff
WALLEYE SPAWN COLLECTION PROCEDURE

1.  Separate ripe males and females into two basins. Keep water fresh.
2.  Prepare the following supplies: a. Two small basins such as oil pans. Rinse with lake water, shake one dry, half fill
  second with lake water.

b. One small bucket. Rinse and half fill with lake water.

c. One large tub (about 18 gallons). Rinse and fill no more than half way with lake
  water. Add clay to make a thin clayey solution. Water should feel slippery. All clay
  should be in suspension. None should be settled on bottom although, TOO MUCH CLAY IS
  BETTER THAN NOT ENOUGH.
3.  Handling fish:
Manually stripping spawn is extremely stressful to fish. BE GENTLE, but also work
  quickly. If either eggs or milt do not flow freely, the fish is not ripe. If eggs are
  clumped, the female is not ripe, never force spawn. If the fish begins to pass blood, stop
spawning immediately.
{A NOTE FROM MTAN} - If eggs are in short supply, you can use "Bloody
  Eggs". The fertility rates may be lower than normal eggs. We suggest that you
  fertilize each pan of eggs using the "Dry" method, then incubate these eggs
  separately from the other eggs you have spawned. Also, if you collect a speared fish that
is ready to spawn, take the eggs!
Hold fish according to photographs (not shown). Grasp the fish near the head between
  your body and arm. With the other hand, grasp the body just above the tail. Hold the fish
  belly down with its back arched over the basin. Press the fish's abdomen firmly beginning
  forward of the vent and working back toward it. Avoid putting pressure too far forward on
the body to prevent damaging the heart or other organs.
{A NOTE FROM MTAN} - we would also suggest that you not apply pressure at the vent
  area as the eggs are expelled. Excessive pressure may tend to break eggs as they leave
  the vent. The protein released from the broken eggs will coagulate and plug the micropyle
of the other eggs, thus reducing the chances of successful egg fertilization. 4.  Spawn one male into the dry small basin. Take care not to allow fish
  slime, blood, or foreign matter to enter the basin. Also try not to get water into the
  basin. EGGS ARE RECEPTIVE TO FERTILIZATION FOR ONLY ABOUT 2 MINUTES AFTER EITHER EGGS
OR MILT BECOME WET.
5.  Spawn females into the dry basin with milt. Spawn 2-3 females per batch.
One batch of fertilized eggs measures about one quart.
6.  Spawn 1-2 more males into the mixture.
7.  Mix spawn by placing the fingers firmly against the bottom of the basin.
Stir without lifting the fingers or touching the sides of the basin.
{A NOTE FROM MTAN} - Again it is important to stress, do not allow your fingers to
  break eggs. Some procedures require the mixing of eggs and milt to be accomplished with a
stiff feather.
8.  Add water from the second small basin. Pour water and spawn back and forth
between the basins 3-4 times.
9.  Pour the spawn and water into the small bucket half full of fresh lake
water. Swirl then let eggs settle.
10.  Pour out half of the water in the small bucket. Add fresh lake water to
the small bucket swirling the eggs in the process. Let the eggs settle.
11.  Pour out 2/3 of the water in the small bucket. Add fresh lake water while
swirling the eggs. Let the eggs settle.
12.  Pour out most of the water. Leave about 1/2" of water covering the
eggs. The water should be clear. If not, continue rinsing.
13.  When the water covering the eggs is clear, pour eggs and water into the
  clay solution. Mix the eggs with the clay solution thoroughly. The clay sticks to the eggs
  and prevents the eggs from sticking to each other while they water harden. Periodically,
remix the egg and clay solution.
14.  Rinse the small basins and bucket with clean lake water.
15.  Let the eggs water harden in the clay solution for at least 5 minutes.
16.  Pour the clay/egg mix through window screen. Rinse the eggs in clean lake
water. Pick out any debris.
17.  Transfer the water hardened eggs to 3/16" mesh screen. Gently rub
  any clumped eggs through the screen. Clumped eggs will eventually die in the incubator
from oxygen deprivation.
18.  Transfer eggs to clean water for transport. The eggs should be no more
than 4-6" deep for transport.
19.  Count the number of eggs per quart. Count the number of quarts of eggs.
  Place eggs in the incubators.  Place water hardened eggs in a 6" trough in a
  single row and count the eggs. Find that number on the D.N.R. Egg Chart that lists the
corresponding number of eggs/ounce and the number of eggs/quart. BIG REDD OPERATING PROCEDURES **Loading eggs into the Big Redd** 1.  Piezometer tube reading = 12 mm. Adjust with piezometer tube air valve.
Record value on "Big Redd Daily Operation Record" forms. 2.  Water exchange rate = 500 ml/minute. Adjust with water valve. Record value
on "Big Redd Daily Operation Record" forms. 3.  Standpipe = High level
4.  Water quality parameters = Record all values on "Big Redd Daily
Operation Record" forms.
5.  Egg volumes: With 1 liter flask, accurately measure the amount placed in
  each tube. Record volume for each tube for each incubator on "Big Redd Production
Record" forms.
**Incubation**
1.  Piezometer tube reading = 25 mm. Egg mass should be at or slightly below
  the lower ends of the tube screens. Record value on "Big Redd Daily Operation
Record" forms.
2.  Water exchange rate = 500 ml/minute or as required to reduce metabolites
to acceptable levels. Record value on "Big Redd Daily Operation Record" forms.
3.  Standpipe = High level.
4.  Water quality parameters - Record all values on "Big Redd Daily
Operation Record" forms.
5.  Airlift screen - Inspect metal screen in airlift assembly when water
  quality parameters are measured. Shut off air valves when removing air lift screen. Eggs
  may rise. Stir eggs GENTLY with metal rod in tank if eggs do not settle when air is
  turned on. Clean airlift screen with water if necessary. Record screen condition on
"Big Redd Daily Operation Record" forms.
6.  Dead egg removal - Siphon off dead eggs (milky white eggs) from tops of
  egg masses. Be careful not to remove living (golden translucent) eggs. Record volume of
  eggs removed from each tube from each incubator on the appropriate "Big Redd
Production Record" form.
7.  Fungicide treatment - Record treatment time on "Big Redd Daily
Operation Record" forms.
8.  Air locks - If an egg mass is lower than that of equal volumes in adjacent
  tubes, then air bubbles have formed beneath the perforated plate of the tube. Plunge tube
  up and down to release bubbles. Record in comments section on "Big Redd Daily
Operation Record" forms.  Water Quality Measure the following parameters EVERY 4 HOURS during Big Redd operation. Follow
instructions specified in each kit or on each meter where applicable. Record results on
"Big Redd Daily Operation Record" forms. Maintain separate "Big Redd"
forms for each incubator. 1.  Dissolved oxygen (DO): With air valves maintain at least 5 ppm (parts per
  million). Measure DO with the Y.S.I. Model 57 - DO meter. Always measure the DO from Tube
No. 1 in each incubator. 2.  Water temperature: Maintain 53 degrees (F). Measure with thermometers
attached to inside wall of each incubator.
3.  pH: Optimal pH is from 6 to 9. Control with rate of water exchange.
  Measure with Cole-Parmer Digi-Sense pH meter model no. 5938-10. Always measure the pH from
Tube No. 1 in each incubator.
4.  Ammonia: Acceptable ammonia levels are unknown. Maintain 5 ppm or lower.
  Control with rate of water exchange. Measure with Hach kit model Ni-8. Always measure the
ammonia content from Tube No. 1 in each incubator.
{A NOTE FROM MTAN} - The book Fish Hatchery Management, published by the U.S.
  Fish and Wildlife Service, suggest that un-ionized ammonia levels should not exceed 0.0125
  ppm. This limit was established for live fish (trout) not eggs. The fraction of total
    ammonia that is toxic ammonia (NH3), can vary greatly
  as changes occur in water temperature, hardness and pH. If water temperatures fall to 41
  degrees (F) and pH levels reach 8.5 then severe ammonia concentrations can develop. Water
  flows through a Big Redd unit are usually high enough so eggs are normally not affected by
ammonia concentrations.
5.  Carbon dioxide: Acceptable CO2 levels are
  unknown. Maintain 5-10 ppm. Control with rate of water exchange. Measure with Hach kit
  model CA-23. Always measure the CO2 content from Tube No.
1 in each incubator.
6.  Water Exchange Rate: Measure rate of flow of water through the Big Redds
  at the outlet with 1 liter graduated flask. Record the flow in milliliters/minute.
  Maintain flow of 500 ml/min except as required to maintain acceptable water quality
parameters. If flow is increased record the changes made.
7.  Piezometer tube: The head of water in the piezometer tube forced above
  tank water level measures air flow. Measure from the tank water level to the bottom of the
  meniscus in the piezometer tube with a metric ruler in millimeters. See sections on each
  operating phase for desired reading. Although, maintaining a DO of 5 ppm takes precedence
over the desired piezometer tube readings.  **Hatching** 1.  Piezometer tube reading = 43 mm unless mass of eggs and/or fry rises above
bottom edge of tube screen. Turn on both air valves. Record on "Big Redd Daily
Operation Record" forms. 2.  Water exchange rate = 500 ml/min or as required to reduce metabolites to
acceptable levels. Record on "Big Redd Daily Operation Record" forms. 3.  Standpipe = High level.
4.  Water quality parameters: Record on "Big Redd Daily Operation
Record" forms.
5.  Egg shell collectors: Put in place (rubber seal down) at hatching. Bottom
  of collector should be just below the bottom edge of the incubator tube screen. As egg
  shells accumulate, the water level in the tubes will rise. When the water level in a tube
  rises above the water level in the tank, remove the collector and rinse with water in the
  white dish pan. Remove and rinse one collector at a time so that any living eggs or fry
  removed with the collector can be replaced into the tube they came from. Replace egg shell
  collectors as quickly as possible. Record time cleaned on "Big Redd Daily Operation
Record" forms.
6.  Foaming: When foaming occurs, attach the J-shaped flexible pipe to the top
of the air lift assembly. The foam will then be directed out of the tank.  **Fry Care** 1.  Piezometer tube reading = 43 mm unless mass of fry rises above tube
screens. Turn on both air valves. Record on "Big Redd Daily Operation Record"
forms. 2.  Water exchange rate = As required to reduce metabolites to acceptable
levels. Record on "Big Redd Daily Operation Record" forms. 3.  Standpipe = High level.
4.  Water quality parameters: Record all values on "Big Redd Daily
Operation Record" forms.
5.  Temperature: Gradually raise water temperature to that of the rearing
ponds. Record temperature on "Big Redd Daily Operation Record" forms.  **Fungicide Treatment** 1.  First treatment is 2-3 days after eggs are placed in the incubator. 2.  Treat every 48 hours thereafter. 3.  Treatment is with 37% formaldehyde (formalin).
4.  If a white precipitate or cloudy suspension is observed in the formalin, DO
NOT USE. The precipitate is paraformaldehyde and is much more toxic than formaldehyde.
5.  Handling Formaldehyde: a.  Wear rubber gloves.

b.  Use formalin only in a ventilated area.

c.  Avoid contact with skin and do not breathe fumes.  If contact is made,
  rinse skin and/or eyes with cold water for at least 15 minutes.

{A NOTE FROM MTAN} - We would strongly urge you to also use eye protection
  (goggles or a full face mask) when mixing formalin, or when moving formalin containers. A
  eye wash station and alarm system should also be set up at the mixing site.
6.a.  Piezometer tube reading = 25 mm

b.  Standpipe = High level

c.  Set formalin dispenser by trial and error to dispense 100 ml water in 3
  minutes.

d. Measure 52 ml water into graduated cylinder.

e. Add 48 ml formaldehyde 37%. This dilution is called formalin.

f. Add formalin to dispenser.

g. Insert dispenser into open top of the operating airlift and dispense fungicide.

h. After 15 minutes remove upper standpipe, drain tank to lower level.

i. Fully open both valves on water manifold.

j. Flush tank for 10 minutes.

k. Replace top section of standpipe. Set for high water level.

l. Fill tank to high water level.

m. Close auxiliary water valve.

n. Record time of fungicide treatment. **Optional Procedures** 1.  Induced Hatching a.  Three days after observing the first fry, increase water temperature 5-6
  degrees (F) for 20 minutes by slowly pouring heated water into the open top of the air
  lift assembly. b. After 20 minutes, allow temperature to return to the former level. 2.  Delayed Hatching a.  Lower water temperature 3 degrees (F) for 1-2 days until hatching is desired.

b.  Return temperature to original level and rapid hatching should occur.
3.  Transporting Fry in the Big Redds***** a.  Operate air pump with deep cycle marine battery and transformer.

b.  Operate standpipe at low level.

c.  Carry ice to maintain desired water temperature. Add as needed to top of
  tubes.

d.  Carry water for metabolite control. Add 2-3 gallons to open top of air
  assembly every 15 minutes.

* Ms. Greiff expressed to MTAN that the above procedure did not work very well
    for her operation. The best way she found to transport fry was in double bagged square
    bottomed plastic bags, which are then placed in cardboard boxes. This procedure involves
  the following steps: a. Add three gallons water to bag.

b. Add two liters of fry.

c. Blow oxygen into the water and partially fill bag with oxygen.

d. Close bag securely with twist tags or rubber bands.

e. Fry will survive for several hours.
Genoa
National Fish Hatchery - Spawning Protocol for Brown TroutBy: Todd TurnerTodd Turner (Assistant Project Leader at the Genoa National Fish Hatchery) has prepared
for MTAN readers, a brown trout spawning protocol which he has found to be very
successful.During the last week of September, the fish are sexed and sorted. Female
fish are anesthetized in 100 ppm of MS 222 and checked for ripeness, beginning in the
first week of October. Once 5% are found to be ripe, spawning commences. The ripe females
are put in a separate holding pen. Two females are stripped into a pan. The males are
anesthetized in 100 ppm of MS 222 and two males are stripped in sequence onto the eggs.
The milt is gently stirred into the eggs. Water from the raceway in added and the mixture
is again gently stirred. The pans are allowed to sit for 90 to 120 seconds. The eggs are
then rinsed, excess water is drained off, and they are placed in 100 ppm betadyne solution
for 30 minutes. The eggs are then placed in a bucket and carried to the broodstock
hatchery building. The eggs are allowed to water harden and are placed in the Heath Trays
at a rate of 3 to 4 quarts per tray. Water flow through each stack of incubators is set at
5 gpm. Eggs are treated daily with formalin at 1:600 concentration for 15 minutes.

Once the eggs have eyed up, they are shocked by dropping them one to two feet into
water. They are then returned to the incubator for 2 days. Three trays from each days egg
take are randomly selected and hand picked. Good and bad eggs are measured and an estimate
of the eye-up for that day is determined. The eggs are shipped within 3 days following
eye-up determination.
Changes/tests 1. Test electronarcosis vs. MS 222 for anesthetizing the fish. 2. Pre-collect milt for same day use to avoid sequencing. 3. Use mechanical egg picker to reduce the number of shipping boxes needed. List Of Fish Hatchery Related
SuppliersBy Mike DonofrioAT the request of MTAN, Mike Donofrio (Tribal Biologist for the Keweenaw Bay Indian
Community) was able to compile a new listing of manufacturers who supply fish hatchery
related products. Mr. Donofrio is presently going through the delightful experience of
dealing with contractors and sub-contractors for the Keweenaw Bay Tribal Fish Hatchery
project. Such an undertaking is a real learning experience. If you find yourself in the
need for some up-to-date "Words of Wisdom", give Mike a call (906-353-6623).
Perhaps you will be able to avoid some of the problems and frustrations Mike has gone
through. Biokyowa, Inc., 930 Roosevelt Parkway, Suite 200, Chesterfield, MO 63017,
    1-800-325-1460, Fry fish food. Industrial Thermoplastics Solutions, P.O. Box 5029, Evansville, IN 47716,  
    1-800-262-4300, PVC fittings and valves. Aquatic Eco-Systems, 2056 Apopka Blvd., Apopka, FL 32703, 1-800-422-3939 (new), General
    Aquaculture. Techno Corporation, P.O. Box 1416,  Erie, PA 16512,  1-814-838-4561,  
    Industrial Check Valves. Nudo Products, 2508 S Grand Ave. E, Springfield, IL 62703, 1-800-826-4132, Fiberglass
    interior boards. Glas-Tek Corp., N6471 Hwy V,  Deerbrook, WI 54424,  1-715-627-4181,  
    Fiberglass tanks. Northstar Company, Route 1, Box 229, Gaston, OR 97119, 1-503-357-4414, Automatic
    feeders. Zeigler Company,  P.O. Box 95,  Gardners, PA 17324,  1-800-841-6800
    (new),  General aquaculture. Northern Lake Services, 400 North Lake Ave., Crandon, WI 54520, 1-715-478-2777, Water
    testing lab. Bioproducts, Inc, P.O. Box 429,  Warrenton, OR 97146, 1-503-861-2256, Fish food. Hatchery TipsBy: MTAN 

Another way to keep yourself out of the Twilight
    Zone is to label, number and perhaps color code your valves and water lines.
    Then write up a protocol which explains the function of each valve and water line system.
    Each protocol should explain the sequence to follow in order to achieve a specific task.Handle all broodfish with care. This will help increase the chance of spawning the same
    fish next year.Once salmonid eggs are placed in the incubators, do not disturb them until they are eyed
    up. Excessive jarring of the incubation tray/jar could result in a lowered survival rates.
    When the eggs are eyed up, proceed with the egg shocking system that works best for your
    operation. Continue to pick dead eggs each day to reduce the chance of fungus (Saprolegnia)
    spreading onto the live eggs.Product and company names mentioned in this publication are for informational
      purposes only. It does not imply endorsement by the MTAN or the U.S. Government. Return to MTAN selection
page Return to Ashland NFWCO home
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