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Hatchery

Technology of
GROUPER

Kingdom: Animalia
Taxonomic
Classification
Phylum: Chordata
Class: Actinopterygii
Order: Perciformes
Family: Serranidae

Overview
Most popular species in the reef food fish industry in AsiaPacific region
Fast growing, hardy ,suitable for intensive culture
Excellent characteristics for processing due to unique culinary
attributes and scarcity
In 1997, the Asia-Pacific Region contributed about 90% to the
world Aquaculture prdxn w/ Grouper at 15000 ton w/ China as
the biggest producer contributing 8000 tons followed closely by
Indonesia. Other countries in the region commonly produces
1000-2000tons annually in 1990-1997.
Although grouper culture is widespread in Asia and Pacific, its
continued dvlp. is constrained by the limited availability of
fingerlings. Most economies w/recent exception of Chinese
Taipei, rely almost totally on wild-caught fry and fingerlings
for stocking.

The word "grouper" comes from the word for the fish,
most widely believed to be from the Portuguese name,
garoupa. The origin of this name in Portuguese is
believed to be from an indigenous South American
language.
In Australia, the name "groper" is used instead of
"grouper" for several species, such as the
Queensland grouper (Epinephelus lanceolatus).

In the Philippines, it is named lapu-lapu in Luzon,


while in the Visayas and Mindanao it goes by the name
pugapo.
In New Zealand, "groper" refers to a type of wreckfish,
Polyprion oxygeneios, which goes by the Mori name
hpuku.

In the Middle East, the fish is known as hammour, and


is widely eaten, especially in the Persian Gulf region.

Groupers are teleosts, typically having a stout body


and a large mouth. They are not built for longdistance, fast swimming. They can be quite large,
and lengths over a meter and weights up to 100kg
They habitually eat fish, octopuses, and crustaceans.
Some species prefer to ambush their prey, while
other species are active predators.
Their mouths and gills form a powerful sucking
system that sucks their prey in from a distance.
They also use their mouths to dig into sand to form
their shelters under big rocks, jetting it out through
their gills. Their gill muscles are so powerful, it is
nearly impossible to pull them out of a cave if they
feel attacked and extend those muscles to lock
themselves in.

Many species move to particular areas at the same time each


year to reproduce in spawning aggregations. In these
aggregations, females release eggs (some larger individuals
produce over 1 million eggs) and these are fertilized by sperm
released by males. The fertilized eggs hatch to very small forms
(larval stages) which drift in ocean currents for 1 to 2 months.
Less than one in every thousand of the small floating forms
survives to settle as a juvenile in shallow water near reefs. As
they grow, they move onto coral reefs and less than one in every
hundred of the young
fish (juveniles) survives to become an adult.

LIFE CYCLE

1-2 months
drifting larval stages

juveniles

Spawning
aggregations in
many species

3-7 years
reef

REPRODUCTIVE
BIOLOGY

Groupers are mostly Protogynous H


ermaphrodites, they mature only as females and
have the ability to change sex after sexual
maturity
Some species of groupers grow about a kilogram
per year and are generally adolescent until they
reach three kilograms, when they become female.
The largest males often control harems
containing three to 15 females.
Groupers often pair spawn, which enables large
males to competitively exclude smaller males
from reproducing.
However, some groupers are gonochoristic.

Gonochorism, or a reproductive strategy with two


distinct sexes, has evolved independently in
groupers at least five times. The evolution of
gonochorism is linked to group spawning high
amounts of habitat cover.
Both group spawning and habitat cover increase
the likelihood of a smaller male to reproduce in
the presence of large males.
Gonochoristic groupers have larger testes than
protogynous groupers (10% of body mass
compared to 1% of body mass), indicating the
evolution of gonochorism increased male grouper
fitness in environments where large males were
unable to competitively exclude small males from
reproducing

PROXIMAL CAUSES
OF SEX CHANGE IN
PROTOGYNOUS
HERMAPHRODITE

CAUSES

Removal of males from the population leads to


sex change in the dominant(largest) females.

The sex ratio would depend upon the current


spawning sex ratio within the aggregation,
the proportion of adults surviving to spawn
in subsequent years, and the proportion of
inshore juveniles surviving to mature and
enter successive aggregations.

Some grouper population, males are more or


less absent till age 10
In case of Red grouper( Epinephalus morio),
the sex ratio is not equal until age 15.
Some Grouper can live

20 years or more!

Fishing

Huntsman and Schaaf (1994) point out that


both protogynous and gonochoristic (separate
sexes) population tends to lose reproductive
capacity as fishing pressure increases.
In protogynous stocks, however, increased fishing
mortality might reduce the relative abudance of
males by reducing the abundance of older-classes
and diminish the probability that eggs will be
successfully fertilized

Loss of behavioural interactions between male and


female

Combined with continued interaction between


females, (Koenig et al. 1999) suggested that
the loss of male-female interaction in Gag
grouper (Mycteroperca microlepis) occur at the
endof aggregation period, when females
purportedly return to swallower inshore
waters.

To date:
Only specie of grouper investigated
Red hind(Epinephalus guttatus) and coney (Epinephalus
fulva) have been histologically proven to be
protogynous hermaphrodites but;
Collin(1992) groupers that aggregate are not necessarily
protogynous.
Ex: Nassau grouper(Epinephalus stiatus) and

Yellowfin grouper( Mycteroperca venenosa),


neither of them shown to be protogynous but
both known to aggregiate.

Nassau grouper (Epinephalus stiatus) spawns in


very large aggregations( thousands to ten
thousands of fish) for only a few weeks of the
year.
In contrast, male and female Gag grouper
(Mycteroperca microlepis) and Red hind
(Epinephalus guttatus) co-occur only during
spawning season.

ARE GROUPERS AVAILABLE ANY TIME OF THE


YEAR?

COLLECTION AREA

MONTH

GROUPER SIZE

Peninsula Malaysia, east


cost

November to December

Average 2cm

Indonesia(e.g., Banten
Bay)

February to April

Fry

Thailand, coastal waters


near mangrove areas

January to March

7.5-10 cm fingerlings

Philippines, variable
peak season in diff.
Province

Peaks mostly in June to


December or October to
November and April to
May

Tinies, fry and


fingerlings

COMMONLY
CULTURED
SPECIES

Brown Marbled Grouper, carpet cod,


flowery cod, blotchy rock cod
Epinephelus fuscoguttatus (Forsskal)
Lapu-lapu

Orange spotted grouper


Epinephelus coloides
LAPU-LAPU

Dusky tail grouper


Epinephelus bleekeri (Vaillant)
Lapu-lapu

HATCHERY
DESIGN

HATCHERY DESIGN

ENTRANCE OF HATCHERY FACILITY

Key

component:
Regardless of
scale is the
implementati
on of
biosecurity to
reduce the
incidence of
disease,
(VNN)viral
nervous
necrosis.

VNN (VIRAL NERVOUS NECROSIS)


A common disease problem in marine finfish
hatcheries affecting most cultured marine finfish
species, including grouper .
Caused by a nodavirus also known as viral
encephalopathy and retinopathy.
Symptoms are disorientation of fish ,change in
color, becoming more dark.
Can be reduced by strict biosecurity. .

KEY FEATURES FOR BIOSECURITY


Separation

of various functional areas;


Access to hatchery limited only to
essential personnel;
Disinfection and thorough cleaning of all
equipments;
Routine monitoring for pathogens and
disease;
Optimisation of water quality and
nutrition to improve overall health and
resistance of larvae.

HATCHERY
OPERATION

BROODSTOCK ACQUISITION
1. Collecting and purchasing of wild fish in a wide
range of sizes.(Epinephelinae, protogynous
hermaphrodites).
In Philippines size of mature female(2.2 kg),and male
(3.5 kg).

2. Grow fish produced in the hatchery (in cage,


pond, or tanks) however, takes 4years to grow
juvenile Tiger Grouper up to broodstock size .

CRITERIA FOR BROODSTOCK


SELECTION
Normal body shape and colour
Absence of skeletal deformities
Overall healthy status i.e., absence of large
wounds haemorrhages, infections, and parasites
Normal behavior, i.e., good response to food
distribution
Best growth and food conversion rate within this
age group.

TRANSPORT REQUIREMENTS FOR


BROODFISH
Transported in dark-coloured, covered tanks
containing aerated or oxygenated water, to
reduce stress.
Dissolved maintained at >75% saturation at all
times.
Mild sedation(using approved sedatives for fish)
to reduce stress and for easier and safer
handling.
Fish should not be fed at least 24 hours before
hand to prevent faeces and regurgitated feed
from fouling transport water.

TREATMENT BEFORE STOCKING

Quarantine the fish


To reduce the opportunity for new fish to transmit parasites
or diseases to the established fish. Quarantine process-takes
between 1and 4 weeks in small tanks(0.5-2 cubic meter)to
facilitate water exchange and fish handling.
This period focuses on reducing parasite load of fish regularly
by placing them in freshwater bath for 5 min. to eliminate
common parasites (skin flukes, protozoans etc.)
If water quality in broodstock tanks markedly different
form previous holding environment, fish acclimation by
placing them in a tank filled with original water, and slowly
add new tank water for up to 1 hr. before releasing them into
the tank.

BROODSTOCK TANKS

Tank size. When fish is(>10kg.) size ranges bet.50-100cubic meters.


Preferred tank color. Medium-range blue, brown or grey not very light or darkshades.
Depth. 2m preferably >2.5 to allow sufficient room for spawning behavior.
Broodstock tanks are roofed in order to reduce the growth of algae on tank walls
making egg collection difficult and increasing egg parasite infestation.
Dirty tanks must be cleaned frequently w/c may stress broodstock and cause
spawning failure or lower quality spawned eggs.
Continously supplied with fresh seawater at daily exchange rate of 200-300%.
Seawater should be filtered and clear with stable salinity (33-35ppt) and water
temperature (27-30.5 degrees celsius).
Tanks located outside are subject to the natural photoperiod, while indoor may be
provided with artificial lightning to simulate different photoperiod regimes.

TANK CLEANING
Siphoning of excessive feed that accumulate on
bottom to prevent water quality degradation.
After spawning remove excess dead eggs w/c
decay and pollute the water .
Broodstock should be bathed in freshwater for 57 min. During tank cleaning to reduce incidence
of parasite infestation.

BROODSTOCK MANAGEMENT
Feeding
feed to satiation (6x a week),w/fish (4x a week) ,w/
squid(2x a week)
Feeding may vary, depending on availability of fish and
squid.{herrings and mackerels are commonly used as feed}
Feed is supplemented with a vitamin mix included at 1% of
feed. Commercial or custom-formulated vitamin mixes can
be used.

COMPONENTS OF FEED MIX


Ingredients

Amount

Minced fish, squid, shrimp, etc.

793g

Rice flour or other finely ground


starch product

195g

Transglutaminase B

10g

Vitamin mix

1-2g(depending on recommended
inclusion rate)

Total

1kg

PROPAGATION

NATURAL SPAWNING
o

Occurs at night 9pm-3am for 3-6 nights


each month during the new moon phase.
However, studies shown that they spawn
throughout the year.
0.8-6.0 M eggs each night.
cease to spawn when temperature drops
around 25 degrees celsius.

IMPORTANT ENVIRONMENTAL
PARAMETERS
ph

7.5-8.3

temperature

25-32 degrees celsius

salinity

20-32 ppt

Dissolved oxygen

4-8 ppm

NO2-N(Nitrogen Nitrite)

0-0.05 ppm

Unionized ammonia

< 0.02 ppm

Australian Center for


International Agricultural
Research ,2012

INDUCED SPAWNING

Prior to Induction: after acquisition of broodfish


is stocked at concrete tanks at 6-12 months;
Stocking density:1/2 cubic meters;
Feed: fed daily with fresh trashfish;
Feeding rate: 5% body weight;
Tank water:50-70% is changed daily.
And condition simulating the natural
environment.

INDUCED SPAWNING CONTINUE


o

o
o
o

Species selected for induction:


female-having mean egg diameter of 400ug.
male- with running milt.
Ratio: female to male is 10:4
Weight range: female(3.6-6.5 kg),male(10-16 kg).
Hormones used: Human Chorionic
Gonadotropin(HCG), Pituitary
Gland(PG),Luteinizing Releasing Hormone a
(LRH-a).
Hormone administration type: Multiple
dosages.

INDUCTION PROCEDURE
First injection, at dosages of 500 IU HCG + 3mg
of PG per kg of fish, and at final injection at 1000
IU HCG + 3mg of PG per kg of fish at an interval
of 24 hours, results showed that treated fish
spawned naturally in spawning tank 12 hours
after final injection.
o However, at lower dosages of 500 IU HCG + 3mg
of PG at 24-hour intervals or using 10 mg LRH-a
at 12-hour intervals, eggs can be artificially
fertilized.
In cases where ovulation did not occur after the
second injection,a third injection was given.

FERTILIZATION
After

injection the fertilized egg that


naturally spawned in tanks were
collected with a fine dip net(100
micron mesh size). Planktonic
organisms and detritus were
removed by screening, unfertilized
eggs settled down on bottom of tanks
were removed by siphoning.

HATCHING
oThe

eggs were placed in


hatching containers and hatched
out in about 15-20 hours.

TABLE 1.TYPE OF HORMONES, DOSAGES AND


TIME INTERVALS USED FOR INDUCED
SPAWNING
Treatment

Hormones

Dosage

Time

used

interval(hr)

HCG + pituitary 500 IU + 3 mg


gland of Chinese PG / kg
carp

12

HCG + pituitary 500 IU + 3 mg


gland of Chinese PG / kg
carp

24

LRH-a

12

HCG + pituitary (1st) 500 IU +


gland of Chinese 3mg /kg
carp
(2nd) 1000 IU +
3mg PG/kg

10ug/kg

24

TABLE 2. INDUCED
SPAWNING OF
GROUPER USING
DIFFERENT
HORMONES

Treatment Fish

Body

Number

Hormone Remarks

number weight

interval of
(hr)

injections

4.5

12

Partial
ovulation, 12
hr after final
injection. No
fertilization.

3.6

12

Partial
ovulation. No
fertilization.

5.2

12

(kg)

Time

used

HCG500
IU+3 mg
PG/kg of
fish

Partial
ovulation, 12
hr after final
injection;
fertilization
rate 40%;
hatching rate
20%. Larvae

Treatment Fish

Body

Time

Number

Hormone Remarks

number weight

interval of
(hr)

injections

4.8

12

Partial
ovulation, 15
hr after final
injection.
Fertilization
rate 30% but
no hatching.

4.2

24

Ovulation 12
hr after final
injection.
Fertilization
rate 60%;
hatching
rate 30%;
larvae

(kg)

used

Treatment Fish

Body

Time

number

weight

interval injections

used

6.0

24

HCG 500
IU + 3 mg
of fish

5.8

24

(kg)

Number of

(hr)

Hormone Remarks

Partial
ovulation.
No
fertilizatio
n.
Ovulation,
15 hr after
final
injection.
Fertilizatio
n rate
30%;
hatching
rate

4.1

12

4.5

12

Partial
ovulation,15
hr after
final
injection.
Fertilization
rate 40%;
hatching
rate 50%;
larvae
healthy.
LRH-a
10u/kg of
fish

Ovulation
12 hr after
final
injection.
Fertilization
rate 80%;
hatching
rate

10

5.2

24

11

5.5

24

12

6.6

24

1st
injectionHCG 500
IU +
3mg /kg of
fish . 2nd
injection
HCG 1000
IU + 3mg
PG/kg of
fish.

All fish
spawned
naturally
12 hr
after final
injection.
Fertilizati
on rate
30%;
hatching
rate 70%,
larvae
very
healthy.

RESULTS FOR INDUCED SPAWNING


The fish under treatment D (Table 1),spawned
naturally in tank 12 hours after the final
injection(Table 2). Fish under treatment B
(Table1) ovulated 12-15 hours after the final
injection, but fertilization was completed only by
artificial stripping.
Fish in treatments A and C required a third
injection for ovulation, eggs were artificially
fertilized 9-15 hours after the final injection.
Ovulation rate of the females treated with
hormonal injections at 24-hour intervals was
higher than those at 12-hour intervals(Table
2),this is due to handling that cause fish stress.

LARVAL AND FRY REARING


(Epinephelus salmoides)
The larvae from hatching containers were collected and
stocked in nine 250-1 fiberglass larval rearing tanks at a
stocking rate of 2500 larvae per tank.
3 types of feed were tested in 3 replicates on a completely
randomized design.
o Type of feed fed to fry:
1st type- sea urchin eggs and Isochrysis;
2nd type-Isochrysis and Brachionus;
3rd type- Tetraselmis and Brachionus
The larvae were reared indoor under intensive care
conditions.
Beginning on day 3 feed was introduced.
20-30% of water was changed daily.

RESULTS FOR LARVAL AND FRY


REARING
sea urchin eggs are suitable feed for grouper
larvae, diameter of sea urchin egg was about
50ug.
Survival rate of fry fed with:
sea urchin eggs and Isochrysis from hatching to
20 days old was 9%;
2% for those fed with Isochrysis and Brachionus;
none of the fry fed with Tetraselmis and
Brachionus survived; all the larvae died after a
culture period of 6 days.

FINGERLING PRODUCTION
At 45 DAH-larvae have metamorphosed into
juveniles ranging from 2.0-2.8cm.
5-40%-survival rates at 45 DAH.
15-25%-survival rates at density of 10larvae/L.
20,000 fingerlings-expected harvest at density of
10 larvae/L, at 10 cubic meter tank.
batch basis in hatchery mgt. that is each batch
of larvae is treated as separate production cycle,
and hatchery is shut down between each
production cycle.

PROPER HANDLING
PRACTICES IN
HATCHERY OF
GROUPER

EGG HANDLING PROCEDURES


Collection
Disinfection
Incubation

Characteristics of Fertilized
eggs

Non-adhesive
Pelagic
Ranges from 0.8-0.9mm diameter

COLLECTION
Fertilized eggs are collected from the overflow of
egg collector tank using fine net.
Removal from collection net once the embryo has
developed optic vesicles.(i.e., at the eyed stage.
However, handling eggs before this stage
increases mortality and deformity.

DISINFECTION
Treated with ozone to minimize the chances of
vertical transmission of VNN (Viral Nervous
Nacrosis) from parent to offspring.
Ozone at concentration of 1 mg/L for 1 min.
Equivalent 0.8mg/L for 1.25 minutes.

Precautionary measure should be taken when


treating Ozone.

INCUBATION
After treatment, eggs are rinsed with clean and
disinfected seawater.
Eggs are transferred into 0.5-1.0 cubic meters
with aerated seawater.

LARVAL REARING PRACTICES AND


REQUIREMENTS
Larval Tanks-volume of 10cubic meter, depth of 1.2m, color
of bright yellow or pale blue to allow grouper larvae to
discriminate prey more easily and tank cleaning easier,
roofed, and enclosed to maintain water temp., and
facilitate biosecurity. Quarantined area with entry and exit
of authorized person only, disinfection of all equipments.
Aeration-provided in a grid pattern to ensure even water
mixture, maintained DO levels, placed in corner to prevent
stagnation, should be in the light in the early stage to
prevent physical damage and could be increased if the
larvae become robust.
Water-should be filtered [filtered through ultraviolet or
ozone disinfection],help maintain biosecurity.

LARVAL REARING PRACTICES AND


REQUIREMENTS
Sea water used must be per-treated using sand
filter to remove particulates, and then sterilized
to reduce potential pathogen intro.
in water.
10 larvae/L-recommended initial stocking density
for grouper.
Oil-added to form thin film on water
surface(around 0.2/m L-meter square)at 1-5 DAH
to prevent surface aggregation mortality in early
stage.

PROBLEMS IN LARVAL REARING


Surface aggregation mortality.
Larvae are attracted to patches of sunlight in the
tank hence, where that patch is present they may
swim to surface of tank. Results in becoming
stock in water and entangled in each other
spines.
Larval mortality at first feeding.

LIVE FOODS
NUTRITIONAL ENHANCEMENT
Larvae of grouper require high levels of highly
unsaturated fatty acids, EPA, ARA, DHA. Provision
via incorporation in live foods.
Recommended for larvae:
Artemia
Brachionus
Isochrysis
Tetraselmis

STATUS AND TRENDS


OF GROUPER IN THE
PHILIPPINES

PHILIPPINES
Grouper aquaculture in the Philippines is based on
the grow-out of wild-caught fry and fingerlings.
Grouper fry and fingerlings are caught using a
variety of methods including hook and line, scoop
or dip nets, traps, gango or fish nest, fish corral, and
several types of nets. In the Philippines, the major
sources of grouper fry include the provinces of
Pangasinan, Cavite, Mindoro, Quezon, Masbate,
Bulacan, Cagayan, Dadiangas, Zamboanga del Sur
and Negros Oriental. The Philippines is one of the
largest suppliers of wild-caught grouper fry, fingerlings
and juveniles in Southeast Asia.

REFERENCES

Australian Center for International


Agricultural Research
SEAFDEC,AQD Iloilo, Philippines
Full- Cycle Aquaculture Status and Trends

NOMU
KAMSAHAMNIDA!!
DOMO
ARIGATO
GOZAIMASU!!