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MATURATION MANUAL
Litopenaeus vannamei
 INDEX

page 3 Introduction

page 4 Biology

page 5 The maturation process

page 6 Maturation building & Equipment

page 8 Biosecurity

page 9 Beginning your new maturation system

page 10 Ablation

page 11 Feed the Vitalis way

page 13 System maintenance

page 14 Sourcing mated females

page 16 Cycling of breeders

page 17 Preparation & daily activities of spawning tanks

page 18 Harvesting nauplii

page 19 Counting nauplii & packing animals for transfer

page 20 Egg & nauplii treatments

page 21 Evaluating nauplii

page 22 Troubleshooting: spawning

page 23 Troubleshooting: maturation

2
INTRODUCTION
DISTINGUISHING FEATURES
The rostrum is armed with dorsal and usually, 2-4 (occasional-
ly, 5-8) ventral teeth, which are moderately long, and in young
distinctly surpassing antennular peduncle. They are shorter in
adults, sometimes reaching only to the mid-length of second an-
Scientific name
tennular segment. Carapace has pronounced antennal and he-
Litopenaeus vannamei patic spines, and lacks orbital and pterygostomian spines. The
Other scientific names
post ocular sulcus is absent. The postrostral carina is of variable
appearing in the length, sometimes almost reaching posterior margin of cara-
literature of this species pace. The adrostral carina and sulcus short, extending to, or only
Penaeus vannamei slightly beyond epigastric tooth. Gastrofrontal carina are absent,
whereas the gastro-orbital carina is relatively short, usually ex-
Common names tending (at most) anteriorly about two-thirds of distance between
West Coast white shrimp hepatic spine and orbital margin. The orbito-antennal sulcus is
Pacific white shrimp well marked, with sharp cervical and hepatic carinae, and deep
King prawn accompanying sulci. Branchiocardiac carina are lacking and lon-
Camaron blanco gitudinal and transverse sutures absent. The sixth abdominal so-
Langoustine mite bears three cicatrices, dorsolateral sulcus extremely narrow
or absent. The telson is unarmed. Antennules lack a parapenaeid
FAO names spine and antennular flagella are much shorter than the carapace.
Whiteleg shrimp The palp of first maxilla is elongate, consisting of 3 or 4 articles,
Crevette pattes blanches with distal ones together flagelliform. The basal article is produced
Camaron patiblanco into setose proximal lobes on the lateral and mesial margins,
which bear 1 or 2 long distomesial spines, and distolateral row of
spinules. Basial and ischial spines are present on first pereopod,
and a basial on second (Perez Farfante and Kensley, 1997).
In mature males, the petasma is symmetrical, semiopen, not hood-
ed, lacking distomedian projections, and has short ventral costae,
not nearly reaching distal margin and distinctly gaping (Perez Far-
fante, 1975; Perez Farfante and Kensley, 1997). The spermato-
phores are extremely complex, consisting of a sperm mass en-
capsulated by a sheath and bearing various attachment structures
(anterior wing, lateral flap, caudal flange, dorsal plate), as well as
adhesive and glutinous materials (Chow et al., 1991).
The mature female has an open thelycum and sternite XIV bear-
ing ridges, prominences, depressions, or grooves (Perez Farfante,
1975; Perez Farfante and Kensley, 1997).
Larval stages: This species has six nauplii stages, three protozeal
stages, and three mysis stages in its life history (Kitani, 1986). The
CL of L. vannamei post larvae range from 0.88 to 3.00 mm (Kitani,
1993). The larval stages (1.95 - 2.73 mm CL) can be recognised by
the lack of a thoracic spine on the 7th sternite, and relative rostral
length against the length of eye plus eye stalk ranges from 2/5 -
3/5, rarely 4/5 (Kitani, 1994). The most distinguishable morphological
character is the development of supraorbital spines in the second
and third protozoea (Kitani, 1986). Coloration: Translucent white,
thus it is most commonly known as the “white shrimp”. The body of
the species often has a bluish hue that is due to a predominance of
blue chromatophores which are concentrated near the margins of
the telson and uropods (Eldred and Hutton, 1960). Size: It grows to
about 230 mm [9 inches] (Dore and Frimodt, 1987).
SIMILAR SPECIES
L. vannamei can be distinguished from L. schmitti, L. setiferus, L.
occidentalis, and L. stylirostris on the basis of the external gen-
italia. In these species, both the thelycum and the petasma, are
more primitive that in species of other genera (Perez Farfante,
1969). The five species of the genus Litopenaeus are restricted
to American waters.
3
 BIOLOGY
Rostrum Adrostral carina

Antennular
flagellum Abdominal segment
Eyestalk

Hepatic spine
Antennal scale

Sixth abdominal
segment
Third maxilliped

Pereiopods
Telson
Antenna Pleopods
Uropod

External anatomy HABITAT

of a shrimp
This marine shrimp likes muddy bottoms at depths from the shore-
Illustration © State
line down to about 72 metres [235 feet] (Dore and Frimodt, 1987).
of Queensland,
Physio-Ecology, Food and Feeding Habits, Reproduction: In
Department of
L. vannamei the carapace is translucent, permitting the colour of the
Primary Industries
ovaries to be seen. In females, the gonad which is first whitish, turns
and Fisheries, 2007
golden brown or greenish brown on the day of spawning (Brown and
Patlan, 1974). The males deposit the spermatophores only on hard-
shell females which will spawn a few hours later.
The cortical reaction is very rapid and first segmentation occurs in
a few minutes (Ogle, 1992). The number of eggs varies according
to individual size. For L. vannamei of 30g to 45g size; 100,000 to
250,000. Eggs are approximately 0.22 mm in diameter. Cleav-
age to the first nauplii stage occurs approximately 14 hours after
spawning (Aquacop, 1979). Toxicity: None. Timing and Meth-
od of Introduction: The Pacific white shrimp was imported in
1985, as post-larvae, by many shrimp farms in South Carolina
(Sandifer et al., 1988).

The courtship and mating behaviour begins in the afternoon in re-
lation to light intensity. Regression of developing ovaries is very rare
and development of the ovaries leads almost every time to spawning.
The spawning process, begins by sudden jumps and active swim-
ming of the female and the whole process lasts about one minute.
DISTRIBUTION
Panama [Golfo de Panama] (Perez Farfante and Kensley, 1997). Dis-
tributed in the eastern Pacific from Sonora, Mexico to Tumbes in
northern Peru (Perez Farfante and Kensley, 1997).

4
THE MATURATION
PROCESS
BRIEF HISTORY
Definition of
maturation The first experimental movements of Penaeid shrimp began in the
early 1970s when French researchers in Tahiti developed tech-
The process niques for intensive breeding and rearing of various alien Penaeid
of conditioning species including P. japonicus, P. monodon and later P. vannamei
broodstock to and P. stylirostris. In the late 1970s and 1980s, P. vannamei and P.
stimulate gonadal stylirostris were transferred from their natural range on the Pacific
development, and coast of Latin America – from Mexico to Peru. From here, they
induce mating, were introduced to the north-western Pacific coast of the Amer-
spawning and icas in the USA and Hawaii, and to the eastern Atlantic coast –
hatching of eggs to from Carolina and Texas in the north through Mexico, Belize, Nic-
produce viable larvae aragua, Colombia, Venezuela and on to Brazil in the south. Most
of these countries have now established the farming of these spe-
cies. Penaeus monodon and P. japonicus were also introduced in
the 1980s and 1990s from Asia to various Latin American coun-
tries and the USA, including Hawaii, (where SPF populations have
been established), Ecuador and Brazil, where introductions were
not successful.
The introduction of P. vannamei to Asia began in 1978/79, when
it was introduced to the Philippines (FAO correspondent), and in
1988 to mainland China (FAO correspondent). Of these first trials,
only mainland China maintained production and started an indus-
try. In 1988, a batch of P. vannamei PL was introduced into main-
land China from the Marine Science Institute of Texas University.
By 1994, Chinese aquaculturists were producing their own PL and
commercial shrimp culture began in the late 1990s. A similar early
introduction of less than 100,000 PL P. vannamei into the Philip-
pines in 1987 from “Agromarina” in Panama was not successful
(Fred Yap, per. com.) and the culture of this species was suspend-
ed for another 10 years.
SPF P. stylirostris have also been experimentally introduced to many
Asian countries (including Brunei, Taiwan Province of China, Myanmar,
Indonesia and Singapore) from secure breeding facilities in Mexico and
the USA. These introductions began in 2000, but have yet to make a
major impact on the culture industries in those countries (with the ex-
ception of a small industry in Brunei), but without notable problems so
far. Penaeus stylirostris was also introduced into Thailand and mainland
China in 2000, but has yet to make much impact in these countries.
(Extract from FAO manual “Shrimp Hatchery Design & Management”).
5
 MATURATION BUILDING & EQUIPMENT
MATURATION BUILDING MATURATION EQUIPMENT

Maturation buildings can be built in many sizes and shapes. The im-
portant characteristic of a maturation building is that it can insulate
the heat from the exterior, making it possible to maintain a stable
water temperature in the maturation tanks.
Buildings are typically constructed of concrete block walls as well as
greenhouses.
MATURATION TANKS
Maturation production tanks can be round, square or rectangular.
The tanks must have a drainage system that allows the water in the
tanks to be drained continuously.
The tanks can be made of concrete, canvas, HDPE etc.

1. Round fibreglass tanks

2. Round plywood tanks
3. Oval tanks made out of block
and HDPE liner
4. Raceways tanks made out of
concrete, block and HDPE liner

2 3 4
6
1 2
1. Recirculating system,
consisting of biofilter,
mechanical filter and
degassing chamber
2. Sand filters
3. Bag filters
RECIRCULATION SYSTEM
A maturation building can be equipped with a recirculation system. This
system usually consists of a biological filter, mechanical filters, bag fil-
ters, a UV filter and a de-gassing chamber to be able to reuse the water
from the maturation tanks. This will decrease the necessity of new water
entering the system as well as increasing the mating efficiency.
SAND FILTERS
Sand filters are rapid sand filters, filled with #20 silica sand, and me-
chanically filter out particulate waste, such as faecal material and un-
eaten feed from the recirculated water.
3
BAG FILTERS
Bag filters are contained in PVC housings. These contain bag filters
down to 25 microns in filtering capacity.
ULTRAVIOLET STERILISER
Ultraviolet sterilisers are used in the recirculating system to disin-
fect the water returning to the maturation tanks.
LIGHTS
Another important criteria for proper shrimp maturation is the photope-
riod. In order for the shrimp to be in full production, you’ll need the days
to be longer than the nights, i.e. 13 hours light and 11 hours dark.
7
 BIOSECURITY
Biosecurity is the implementation of measures to avoid the entrance The following is a summary of the biosecurity risks that should be
of infection into a unit, control the dissemination of an infection within reviewed within all production units to identify and minimise the risk of
the unit, and avoid the spread of infection to other units. diseases entering the unit:

SITE LOCATION PEOPLE WATER INTAKE SUPPLIES INTRODUCTION DISPATCH A QUARANTINE CARRIERS
MOVEMENT AND OF GENES AND LOADING AREA IS
BEHAVIOUR · Fresh/frozen PROCEDURES RECOMMENDED, · Wild and
feed sources Broodstock where the animals domestic animals
· Staff are held and
· Other supplies · Pests
· Visitors analysed before
entering the
production area.

8
BEGINNING YOUR NEW MATURATION SYSTEM
INTRODUCING NEW BROODSTOCK

NUMBER OF SHRIMP YOU NEED IN You want shrimp that are in the 40g range. This depends on the
YOUR MATURATION SYSTEM quality and performance history of the animals. Some broodstock
producers have been systematically breeding for maturation of the
DENSITIES AND POPULATIONS animals to begin at as small as 30g. As a general rule, 35g males and
40g females are the optimal size for maturation, as the majority of the
Maturation tanks should be stocked at 8-10 shrimp per square me- male and female sexual organs will be fully developed by this stage
tre. Production tanks are stocked with a 1:1 ratio of males to females. of growth. Also look at the spermatophores in the males – make sure
that they are well developed and that there is no evidence of melani-
Note that the recirculation system must be able to handle this density sation on either the spermatophore or petasma.
of animals.
Maturation STOCKING MATURATION TANKS
parameters If you do not use a mixed gender system, stock your female tanks
and male tanks at a density of 8-10 broodstock per square metre. Upon the arrival of new animals for maturation, calculate the number of an-
The water parameters imals needed to stock in each maturation tank to achieve the desired den-
of your maturation MAKING SURE YOUR MATURATION sity. Stock an additional 10% to compensate for post-stocking mortalities.
tanks should be: SYSTEM IS CLEAN
· 30-35 ppt. But it must Divide shrimp into maturation tanks as evenly as possible, separating by
be constant. Broodstock must be replaced every four months, after ablation of sex as much as possible. This will allow you to ablate females without
the animals. having to sort through males. After the populations have stabilised, fe-
· Temperature of males ablated, and mortalities made up, the sexes can be mixed. Leftover
the water must be The tank and filter system must be periodically cleaned and disin- shrimp should be kept in a tank and used as replacements for mortalities.
28°C +/- 1°C fected to avoid sponges, bacteria, protozoa bryozoans and other
organisms to foul the tank wall and the PVC pipes. ACCLIMATION
· Photo period of
13 hours light and If a cleaning protocol is not followed this will cause the water quality Acclimation is dependent on the parameters of the water when the
11 hours dark to decline and the efficiency of the maturation will also decrease. broodstock arrives. A basic guideline to use is not to raise tempera-
ture more than 4°C per hour and no greater than a 10% change in
· Light intensity
PREPARING MATURATION TANKS salinity per hour. The purpose is to reduce the amount of stress that
subdued 12
new animals experience when they are placed in a new environment.
micro watt/cm2
Set up large 15cm standpipes over the 5cm standpipes to keep
weak shrimp from being sucked into the pump. These can be re- However, even though parameters match there should still be an ac-
moved after one week. climation process, as you do not know the exact chemistry of the
seawater and acclimating strictly for temperature and salinity may not
Test the system for residual chlorine and make sure all pumps and be enough. Adding water from our system to the bags helps tremen-
filters are functioning properly. Salinity should be 32ppt and tempera- dously in reducing the stress on the new animals. Transfer the bags of
tures should be 28°C. animals directly into the tanks that will be used for the production run.

9
 ABLATION
Generally it is done no sooner than 10 days after the shipment has
arrived. Guidelines: (1) most of the animals have molted; (2) the exo-
skeletons are hard; and (3) shrimp are not exhibiting signs of stress.
The process of ablation is the removal of one of the glands in the eye-
stalk of the female shrimp. It is a process called unilateral nucleation.
Hold the shrimp, tail folded to meet the head, and isolate one of the
eyes while restraining the female. Using red-hot scissors, remove the
right eye of the females at the base of the eye. Make sure that you
remove the entire eyestalk and the eye because the eyestalk contains
an organ near its base that regulates ovarian development.
It is best to give the females a few days to recuperate from the stress
of ablation before sourcing the animals. After about a week the ani-
10
2
1. Preparing the scissors
for unilateral nucleation
mals should be strong enough to be handled and sourcing can be- 2. Ablating the right eye of
the female
gin. Production will not be that strong for a couple of weeks but it
should pick up by about week three.
At present, unablated females are being used in some matura-
tions around the world. This has contributed to increased animal
welfare and the production results are comparative with ablated
females.
If you use a mixed tank system, place the males and females in the
tanks at a ratio of 1:1.
If you do not use a mixed system, place the females in the tanks as
well as the males at a density of 8-10 broodstock per square metre.
FEED THE
VITALIS WAY VITALIS

One of the most important factors in maturation is feed. High-qual-

ity feed is important for maturation. The feed types and freshness Vitalis 2.5 is a
can mean the difference between 50,000 nauplii and 300,000 nau- cold-extruded
plii per female. Seasonality is a challenge in receiving the same semi-soft pellet
quality feeds year round. Vitalis 2.5 replaces part of these fresh
It is formulated and
feeds, and as Vitalis 2.5 does not depend on seasonality, produc-
produced at the
tion is standardised year round. It is recommended that you start
Skretting centre
incorporating Vitalis 2.5 to your traditional protocol at 3%. Feed
of excellence for
your maturation for two weeks for the animals to get used to the
hatchery diets in
new feed. After which, start decreasing one of the fresh feeds until
France. The formula
you have the production numbers you are looking for. You can fol-
is based on a high
low this process with several of the fresh feeds.
degree of marine
protein components,
algae, omega-3 fatty
acids (DHA/EPA),
vitamins and minerals.
The diet is formulated
with a high protein
content to support
the high fecundity &
spawning frequency
and is 2.5mm
in diameter and
approximately 5mm in
length.

1. Weighing bloodworms
1 2 2. Vitalis 2.5

11
 FEEDING RATES
DETERMINING FEEDING RATES
As a guideline, the shrimp broodstock should be fed about 26% to
30% of their biomass per day on a wet weight basis. Feed consump-
tion will vary with the mating frequency and growth rates of the an-
imals. Begin feeding the animals at 26% of the total biomass and
increase or decrease the feed amounts according to actual feed con-
sumption and nauplii production.
% wet bodyweight
Bloodworms 3-10%
Squid 5-10%
Artemia biomass 3-6%
Mussels or clams 3-6%
Vitalis 2.5 3%
Feed 6-8 times over 24 hours
Feed what is available per region
Vitalis 2.5 must be divided in 6 feeding times.
12
PREPARING FEED SHEETS
The feed sheet is generated using a spreadsheet that should be print-
ed in colour to make it easier to view the different rations. It should
show the individual rations and when the animals should be fed. Note:
Look at Excel sheet for AquaSim.
FEED ORDERING
Order feed for a minimum of two months and up to four months at a
time. You do not want to keep too much feed to hand because it can
still spoil, but you also do not want to run out.
FEED STORAGE
All feeds should be stored in a freezer located in the maturation lab.
The inventory must be rotated when a new shipment arrives so that
the oldest feed is used before the newer feeds.
FEED PREPARATION
Look at the feed sheet to determine how much feed is going to be Squid should allow
needed, and remove the appropriate amount from the freezer. to sit out for a few
minutes, but must
Bloodworms should be allowed to defrost, then the ration is weighed not be defrosted.
Once it has warmed
out and distributed into individual feed cups.
up slightly and is no
The Artemia is fed frozen. Using a sharp knife, it can be scored and longer rock solid
broken like a tile. The blocks of frozen Artemia are added to the tanks it can be cut into
and allowed to melt as they circulate around the tanks. The pumps 5cm square pieces.
should be shut off during Artemia feeding to allow the Artemia to thaw After that, weigh out
and drop to the bottom of the tank. If pumps are left on, the floating the ration for each
block will move towards the centre of the tank and pass down the individual tank.
centre standpipe.
 SYSTEM MAINTENANCE
FEED STATION
The feed station is where all of the foods are prepared and need to
be kept clean at all times. Cutting implements, surfaces, and feed
containers need to be disinfected with bleach and left to dry. Keeping
things clean ensures the contamination levels are low.
CLEAN UP
All equipment needs to be cleaned and disinfected using bleach, then
set out to dry. A chlorine bath should be set up at the feed station for
sterilising equipment and feed cups. The chlorine bath should be pre-
pared at a concentration of 540ppm of active ingredient.
FEEDING PROTOCOL
Using Vitalis 2.5, you will be able to replace a big portion of the fresh
feeds. Depending on the shrimp genetic strain you will be able to re-
place from 20% up to 70% of the fresh feeds with Vitalis 2.5.
Cutting Artemia
biomass
The importance of keeping the water and filters clean cannot be
stressed enough. Failure to keep the systems clean can lead to bac-
terial problems, algae blooms, high levels of particulate in the water
column, fungus and a host of other organisms that could develop into
a serious problem for production.
SIPHONING
To construct a siphon, use 2.54cm PVC and connect this to rigid
1.095cm ID-reinforced vinyl tubing. Maturation tanks should be si-
phoned when there is a build-up of detritus or feed particles that are
too small for the molt nets to remove from the maturation tanks. Si-
phoning is also required when there is a build-up of sand in the tanks
from broken laterals in the sand filter.
Another method is to daily use a hand net to remove any excess feed.
MOLT REMOVAL
Uneaten food, dead shrimp and molts should be removed from the
maturation tanks on a daily basis. This is an important step in keeping
the maturation tanks and the water as clean as possible.
Incoming air should be shut off to reduce turbulence in the water. A
mesh net is pushed through the tank slowly with constant forward
momentum to keep debris from coming out of the net and removing
waste from the bottom of the tank. Once all of the waste has been re-
moved from the tank it is transferred to a collection bucket for removal
after all of the tanks have been cleaned.
13
 SOURCING MATED FEMALES
Each day at 18:30-19:00 (1 hour after the lights are switched off),
mated females should be removed from production tanks and placed
into spawning tanks. The tip of a fishing rod is used to help turn fe-
males enough to see if they have a spermatophore attached. Mated
females are netted out of the tank and lifted out of the net and held
with their tails tucked under the abdomen. This position prevents
14
them from flapping their tails and possibly dislodging the spermato-
phore. The mated females are quickly carried over to the spawning
tank where they are gently released. A tally should be kept to keep 1. Mating behaviour (male
track of how many females came out of each production tank so that chasing a female)
the same number of females will be returned to that tank after they 2. More females ready to
have spawned. Sourcing generally takes one hour per system. be sourced and mated
2
RETURNING FEMALES

At 03:00, spawning tanks should be checked to see if females have spawned. If all the females have spawned, then you can begin returning
females to production tanks. If all females have not spawned, they are left alone and checked later. Once all females have spawned they are 3. Sourcing mated females
netted and released into production tanks based on the tally (taken earlier while sourcing). No more than two shrimp are carried in the net 4. Female already sourced
and once the shrimp are in the net, they are flipped over to prevent them from flopping. with a spermatophore
attached

3 4

15
 Spawning tank with egg
collectors on the bottom for
easy harvest

CYCLING OF BREEDERS Use of Vitalis 2.5

The use of Vitalis 2.5

has also proven to
Animals are maintained in the system for approximately five months. The broodstock will continue to produce acceptable numbers of keep the broodstock
Subtracting the two weeks spent in the conditioning and post-abla- nauplii for longer periods of time if the animals are well-nourished, healthier for a longer
tion, then another two weeks for culling the animals and preparing the systems are kept clean, and if the breeding populations are main- period of time.
the system for new animals, and your populations are in production tained at the right densities.
for approximately four months.

16
PREPARATION AND DAILY ACTIVITIES
OF SPAWNING TANKS
One spawning tank is used for all the mated females from each maturation system. The ratio of spawners in a spawning tank is two spawners
for every 100 litres of water. This method of spawning is called mass spawning and is preferred over having females in individual spawning
tanks because of the labour involved.

Spawning tanks

Spawning tanks are

filled with 1 metre of PREPARATION AND FILLING OF SPAWNING TANKS
sea water at 30-34ppt
salinity and maintained CLEANING TANKS
at a temperature of
29-30°C. Before each use, the spawning tanks must be carefully cleaned and
disinfected. Cleaning the tanks consists of a series of steps that are
important for proper disinfection:
1. Rinse tanks to remove eggs that have not hatched and other
organic debris left behind after the harvest.
2. Mix bleach and water mixture, making sure that it is well mixed
to assure proper disinfection.
3. Scrub the bottom of the tank and sides using the bleach solution
in the tanks.
4. Drain the tanks and squeegee out excess water and eggs from
the tank.
5. Using a bucket filled with straight bleach, scrub the bottom and
the walls of the spawning tank using the broom.
6, Rinse the tanks out twice using fresh water.
7. Rinse the tank a third time using salt water and squeegee out
the excess water.
8. Clean the airstones.
9. The spawning tanks are now ready to be filled.
FILTRATION OF SPAWNING TANK WATER
Normal seawater could be used for hatching, but for optimum hatch-
ing rates we polish the water and adjust the salinity. Before being col-
lected in a 50-tonne reservoir, the water used for spawning passes
through filter bags of 1 micron.
FILLING THE SPAWNING TANKS
The spawning tanks are filled the morning after they have been
cleaned. Place the graduated standpipe into the tank and begin
filling the spawning tanks with filtered seawater from the spawning
tank reservoir. The spawning tanks should be filled to a depth of
approximately 1 metre with seawater (approximately 33ppt).
SPAWNING TANK TREATMENTS
Once all females have been returned and the heater is placed in
the spawning tank, the eggs can be treated with 5ppb Treflan. After
treatments, eggs are stirred using a squeegee. The stirring pattern
is three strokes to the right, three strokes to the left and then three
strokes in the middle.
After the eggs are stirred, the airstones are placed in the tank
and adjusted to provide moderate aeration. Before leaving for the
night, all tanks are checked to make sure heater is in place and
if needed, treatments have been added and airstones are in and
properly adjusted.

17
 HARVESTING NAUPLII

1 2

1. Harvesting the nauplii

MIXING THE NAUPLII FROM THE EGG CLUSTERS
The first job of the morning for the spawning person is to tend to the
row of spawning tanks and stir each to break up the eggs and help
free the nauplii from the bottom of the tank.
from the spawning
tanks using the nauplii
SETTING UP THE NAUPLII COLLECTORS collectors
2. Nauplii ready for harvest
Next, they should go through the spawning tanks and set up the nau-
plii collectors for harvesting. Place harvest barrels inside outer jackets
and attach the hose from spawning tank to the harvest box. Add about
20cm of water to harvest barrel so that when the nauplii harvest be-
gins, the nauplii are not slamming into the bottom of the harvest box.

18

The nauplii are
harvested using light.
The strongest nauplii will
be attracted to the light
as they have a positive
phototactic response.
Only the strong nauplii
are harvested and
transferred to the larval
production areas.
TRANSFERRING NAUPLII INTO
THE NAUPLII TANKS
After nauplii are harvested from the spawning tanks they are concen-
trated in the nauplii collectors and transferred to nauplii holding tanks.
The nauplii remain in these tanks before being harvested and trans-
ferred to larval rearing.
COUNTING NAUPLII &
PACKING ANIMALS
FOR TRANSFER
To get a count of the daily production you need to sample the nauplii
tanks and extrapolate to figure out the number of nauplii in the tank.
The tanks are approximately 500 litres.
1. Take three samples using a 1ml pipette.
2. Place each sample in a labelled 35ml plastic beaker.
3. Take samples to the lab for counting. Place the sample to be
counted into a petri dish, fill a beaker with freshwater and pour
into the petri dish to kill the nauplii. Nauplii are counted as they
are sucked up into a pipette.
4. Samples are counted individually and if the standard deviation be-
tween the three samples is less than or equal to 10%, these num-
bers can be used for your calculations. If there is too much varia-
tion between the counts then two more samples are counted. Out
of the five samples counted, the high and low are eliminated and
the other three counts are used for population calculations.
5. Average the three numbers and multiply the average by 14,400.
Since the samples are 5ml, multiplying by 200 yields the num-
ber of nauplii per litre and since there are 72 litres in a harvest
barrel multiplying by 72 provides the number of nauplii in a har-
vest barrel.
6. Record the data on the “daily nauplii count” sheet and “transfer log”.
7. Take the completed transfer log sheet to the hatchery and have one
of the hatchery managers to fill it out. They will dictate how many
and which nauplii will be stocked into which larval rearing tanks.
Once the sheet has been filled out by the appropriate hatchery per-
sonnel, nauplii are consolidated in harvest barrels as per their request.
19
 EGG AND NAUPLII TREATMENTS
There are many different treatments used to disinfect eggs and nau-
plii. Treatments vary with respect to the types of chemicals that are
used, dosages and treatment times.

The following are some treatments used to disinfect eggs and nauplii:

EGG TREATMENTS

50ppm Argentyne for 60 seconds

20ppm Argentyne for 5 minutes

N-3 TREATMENT

25ppm Argentyne for 3 minutes

N-5 TREATMENTS

50ppm Argentyne for 60 seconds

25ppm Argentyne for 3 minutes

2ppm Chloramine-T for 5 minutes

Nauplii

20
EVALUATING NAUPLII
This can be a valuable quality control tool. By looking at your animals on a daily basis you will be able to verify how changes in the environment
and management can affect the quality of your animals. A sample of nauplii from each system is checked daily. A grade is given to each system
from microscopic observations based on melanisation, necrosis, nauplii stage and deformities.

Nauplii

21
 TROUBLESHOOTING: SPAWNING
The following highlights some of the more common problems that have been observed in the spawning area, their cause and recommended solutions.
AREA CAUSE
Plugged concentrator screens Dirty screens. Water circulation pattern into
concentrator not set properly. Too many nauplii in a
concentrator. Eggs accidentally harvested.
Eggs settling out in a pile in the centre of the spawning Tank was stirred improperly, pattern was not followed.
tank
Nauplii outside the concentrator during harvest Nauplii were spilled while setting up for harvest. The
concentrator screens were not checked before harvest
was started. A screen patch came undone or a new
hole developed during harvest.
Nauplii not coming to the surface Drop in atmospheric pressure due to storms or other
weather conditions.
Nauplii with broken appendages Animals are getting caught in the screens of the
concentrators.
Severe deformities Animals naturally deformed. Make sure EDTA Versene levels are correct, increase
22
SOLUTION
Clean screen with sponge. Set 2.54cm hose in the
airline placement provided for each concentrator.
Do not harvest more than 4 million nauplii into a
concentrator. Take care that eggs have settled out
after you stir them.
Follow correct stirring pattern, use the tickle stick to
stir the pile into the water column.
Be aware not to spill nauplii outside the concentrator.
Be conscious and check every concentrator screen
before starting the harvest. If a small hole is found,
repair with a dab of silicone. Large holes should
be repaired by replacing the entire screen to the
concentrator.
Check all water parameters as well as temperature
parameters.
Check that the screens are clean, make sure that the
harvest hose is set up correctly, and that the person
handling the nauplii is being careful and deliberate
about his or her job.
the level by 5ppm, change the diet, double check all
parameters of the spawning tanks.
TROUBLESHOOTING: MATURATION

Nauplii

Vitalis 2.5

Problems in production will arise from time to time and are not always systems are maintained at 28°C +/- 1ºC. Drops in the temperature The use of Vitalis 2.5
easy to solve. Potential problems that may be experienced are listed will slow mating and ovarian development. Increases in tank tem- delivers a consistent
below, along with possible causes and solutions. Remember every peratures may cause algae blooms or bacterial problems. Prolonged production to the
case is different and needs to be treated as such. high or low temperatures will acclimate the animals to a new tem- maturation department,
perature, so switching the animals back to the prescribed tempera- as the ingredients/
POOR QUALITY BROODSTOCK tures will cause erratic production until the animals acclimate back to nutrients delivered
the proper temperature. through Vitalis 2.5
Once you are sure the animals will not perform to your standards and are always constant,
cannot effectively be worked with, the group should be eliminated as FEED thereby allowing the
soon as it can be replaced. Current production standards are at least maturation department
10% of the females spawning and production of at least 150,000 nau- Feed types, quality, ration (amount of feed) and freshness are all import- to produce a consistent
plii per female. ant in maturation. Poor quality food due to improper storage, handling or amount and quality of
preparation can cause problems for your animals and reduce production. nauplii year round.
TANK TEMPERATURE No more than a three-month supply of feed should be ordered at a time.

Tank temperatures are known to affect mating frequency. For consis- If the ovaries are not developing, look to increase the amount of poly-
tent production, the tank temperatures need to be stable. Production chaetes. If the males are not responding, increase the squid.

23
 Maturation tanks

Hatching
SOURCING ANIMALS TOO SOON OR LATE the heat exchanger, aeration, EDTA levels, nitrites and whether or not temperatures
the egg count looks low. It must be determined whether it is a hatch-
The idea of the controlled photoperiod is not to get all of the shrimp ing problem or low egg production in the females. If the water in the
that will mate and spawn per day. Instead, the objective is to allow spawning tanks
enough time before the tank is sourced so that the majority of the grav- MISMATES falls outside of the
id females will have mated but not spawned. Although you want all the optimum hatching
gravid females to be mated by the time you source, it is likely that some Mismates are females that have a spermatophore attached but not temperatures you will
of them will still not have mated by that time. Still, look to maximise. placed properly. Mismates may be caused by not having enough see deformities, low
water depth in the maturation tanks or by turbulence in the water. nauplii yields or no
Make notes concerning mating while sourcing. If you are finding Increase the depth of water in the maturation tanks to give the males hatches at all. Maintain
mated females that have no development, it is most likely they have more room to chase and mate with females. Another possible rem- the spawning tanks
already spawned and you need to adjust the sourcing schedule. If, edy is to shut the pumps off earlier to reduce the current and turbu- at 28°C-30ºC and
when you arrive to source, there is still courtship activity going on lence in the water. If all else fails, look for new males. make sure that tanks
and a high percentage of gravid females have still not mated, then temperatures are kept
you may be sourcing too early. In this case, you may want to give the ABORTS between 28°C and
animals more time to mate. 30°C or you will run
If the water in the spawning tanks is too cold or the females are dis- into problems.
LOW NAUPLII YIELDS turbed while spawning they will abort. Excessive handling or rough
handling of the females during sourcing is another major factor leading
Low nauplii yield can usually be attributed to nutrition (as mentioned to aborted spawns. Cold temperatures, stress and new production
previously) or to problems in the spawning tanks. You need to check animals are the major causes of aborts. Adjust the thermostats on the
all of the parameters in the spawning tanks prior to stocking and just heat exchanger to make sure the water temperatures are maintained
before harvest to make sure that nothing is out of the ordinary. Check at 29ºC-30ºC. Also, try not to disturb the spawning females.

24
 BROODSTOCK MORTALITIES

Problems with mortalities are not always within your control, but there are
a few things that you can make sure of is that management is not contrib-
uting to the problem. The normal mortality rate once the populations have
stabilised is between 0.1 and 1.0% of the population per day.

The following are some of the common causes of broodstock mortal-

ity, and some potential remedies:
1. Acclimation problems

If the acclimation procedure is not done correctly, you can lose all of the
animals. Make sure to match all of the water parameters both physical
and chemical as closely as possible. Any differences in these parameters
need to be changed gradually to minimise the stress on the animals.
2. Water quality

Good water quality is essential for having a good productive system.

Ammonia and nitrite levels are checked weekly to make sure biofilters
are functioning properly. Tank temperatures for production systems
and spawning tanks are taken daily to ensure that they are within ac-
ceptable levels. Dissolved oxygen and pH levels are checked if a prob-
lem is suspected. Bacteria levels are checked before and after UVs
on a weekly basis. Given the large volume of the maturation system,
it takes a couple of days to completely flush the system out. Since it
takes a couple of days to replace the water you need to be careful
with bacterial levels and blooms of toxic algae.
3. Diseases

At the first sign of possible diseases (lesions, expanded chromato-

phores, black legs) have health assurance check broodstock for bac-
teria. If excessive mortalities are seen, consider reducing densities and
increase your exchange rates. If necessary, tissue samples can be
sent out for further testing for viruses.
4. Handling

Always remind personnel that the shrimp are fragile and need to be
handled gently. Shrimp should be held in a jackknife position to keep
them from kicking and injuring themselves. Never catch more than
one animal at a time when sourcing, and do not transport more than
Nauplii tanks
two animals in a net at a time. When transporting these many animals
you need to immobilise them to keep them from injuring one another.

25
 KEYWORD EFFECTIVENESS INDEX

Ablation .......................................................................................... pages 9, 10, 16 Feed Preparation ......................................................... page 12 Preparing the Maturation Tanks ........ page 9
Aborts .................................................................................................. page 24 Feed Sheets ........................................................................... page 12 Recirculation System .......................................... page 7, 9
Acclimation ............................................................................ pages 9, 25 Feed Station ........................................................................... page 13 Returning Females ................................................... page 15
Acclimation Problems ....................................... page 25 Feed Storage ....................................................................... page 12 Sand Filters .............................................................................. page 7
Anatomy ......................................................................................... page 4 Feed Types ............................................................................. pages 11, 23 Scientific Name ............................................................... page 3
Bag Filters ................................................................................... page 7 Feeding Rates .................................................................... page 12 Setting up the nauplii collectors ........... page 18
Beginning ..................................................................................... page 9 Filling the Spawning Tanks ..................... page 17 Similar Species ................................................................. page 3
Biology ................................................................................................ page 4 Filtration of Spawning ........................................ page 17 Siphoning ..................................................................................... page 13
Biosecurity ................................................................................. page 8 Habitat ................................................................................................ page 4 Sourcing Animals
Broodstock Mortalities ..................................... page 25 Handling ....................................................................................... pages 22, 23, 24, 25 Too Soon or Late ......................................................... page 24
Buildings ......................................................................................... page 6 Harvesting Nauplii ...................................................... page 18 Sourcing Mated Females ........................ pages 14, 15
Cleaning Tanks ................................................................. page 17 Hatching Temperatures ................................. page 24 Spawning Tank Treatments ................... page 17
Common Name ............................................................. page 3 History .............................................................................................. pages 3, 5, 9 Spawning Tanks ......................................................... pages 14, 15, 17, 19,
Counting Nauplii ............................................................ page 19 Larval Stages ....................................................................... page 3 22, 24, 25
Cycling of Breeders ................................................ page 16 Lights.................................................................................................. pages 7, 14, 22 Stocking Maturation Tanks .................... page 9
Daily Activities of Spawning .................. page 17 Low Nauplii Yields ..................................................... page 24 System Maintenance .......................................... page 13
Definition of Maturation .................................. page 5 Maturation Building .............................................. pages 6, 7 Tank Temperature .................................................... pages 23, 25
Densities and Populations ....................... page 9 Maturation Process ................................................ page 5 Transfer ............................................................................................. page 19
Diseases ...................................................................................... pages 8, 25 Mismates ....................................................................................... page 24 Troubleshooting: Maturation ....................... page 23
Distinguishing Features .................................. page 3 Mixing Nauplii and Egg Clusters ............ page 18 Troubleshooting: Spawning ....................... page 22
Distribution ................................................................................. page 4 Molt Removal ....................................................................... page 13 Ultraviolet Steriliser ................................................... page 7
Egg and Nauplii Treatment ..................... page 20 Number of Shrimp .................................................... page 9 Vannamei ...................................................................................... page 1, 3, 4, 5
Equipment ............................................................................... pages 6, 13 Packing Animals for Transfer ............. page 19 Water Quality ........................................................................ page 9, 25
Evaluating Nauplii ........................................................ page 20 Parameters ............................................................................ pages 9, 22, 24, 25
Feed ......................................................................................................... page 7, 11, 12, 13, 23 Poor Quality Broodstock ............................ page 23
Feed Ordering .................................................................... page 12 Preparation of Spawning Tanks ........ page 17

26
OUR SHRIMP HATCHERY DIETS ARE PRODUCED AT OUR CENTER OF EXCELLENCE IN FRANCE
AND SOLD IN MANY COUNTRIES IN ASIA AND SOUTH AMERICA

For more information, please contact Aedrian Ortiz, Skretting Marine Hatchery Feeds Technical Support Manager
Tel: +52 812 351 2286 - E-mail: aedrian.ortiz@skretting.com

© All rights reserved. This document has been prepared by and belongs to Skretting. No partial or complete copy or use of this document is allowed without the authorisation of the author.
Whereas the latter ensures that the information is correct, he cannot be responsible for potential incorrect information.
Information provided in this manual is a guidance therefore Skretting cannot be responsible for results arising from the use of it.
Farming practices in the hatcheries should comply with all local farming regulations and standards.