Experience Work on Development Of Broodstock Management And

Sustainable Mass Seed Production Management System In Giant Grouper at FRI Tg Demong

Nik Haiha Nik Yusof, Ahmad Daud Om, Sufian Mustafa, Kho Li Young, Shaharah Md Idris and Nik Daud Nik Sin

Fisheries Research Institute Tanjong Demong22200, Besut, Terengganu, MALAYSIA

AQUACULTURE OF GIANT GROUPER

Giant grouper is one of the highly esteemed coral reef food fish in the East Asia region especially Hong Kong and China.

Currently, only Taiwan claimed successful in closed cycle captive breeding of giant grouper commercially.

  In Malaysia, export demand for giant grouper is

increasing and the fish priced at RM 40.00-60.00/kg ex-farm.

Price of an imported seed of 5.0-10.0 cm TL is ranging from RM 5.00 – 10.00 per tail inclusive transportation cost.

MAJOR ISSUES IN GIANT GROUPER CAPTIVE BREEDING

A giant size broodstock require bigger and deeper broodstock-cum-spawning tanks either on land or at the sea. Therefore will pose a great constraint to keep the broodstock under control environment for year round spawning.

Late maturation of about 5-7 year, therefore require extra holding facilities for keeping of stand-by young and matured broodstock.

Protogynus hermaphroditic species, meaning that most of the fish matured as females and later transformed into males. Thus create imbalance ratio between males and females.

Lack of males naturally will affect consistency production of good quality eggs through the year.

Lack of natural spawning will increase mortality risk on the broodstock due to excessive handling. Production cost will also be higher due to a huge requirement of hormones for induce sex reversal and spawning.

Low survival rate at various production stages due to mortalities related to initial feeding, shock died syndrome and other diseases.

High cannibalism activities especially during metamorphosis until the fish attained 10.0-12.0 cm in total length (TL). This will reduce final survival rate significantly.

CHRONOLOGY COLLABORATIVE R & D ON GG

• Joint Research and Development Project on Breeding Biology and Artificial Seed Production of Giant Grouper (Epinephelus lanceolatus was collectively agreed by Malaysia and China during The 2nd Meeting of Malaysia-China Joint Working Group on Agricultural Cooperation in Beijing, China on the 23rd – 25th May 2005. The project is planned concurrently for five years beginning 2005 to 2010 and will be jointly carried-out by Marine Finfish Fry Production and Research Centre (MAFPREC), Tanjung Demong, Besut, Terengganu, Malaysia and Guangdong Dayawan Fishery Development Centre (GDFDC), Yaqian, Aotou, Huizhaou City, Guangdong Province, People Republic of China. (does not take place)

• Research collaboration in development of seed production of grouper (Ephinephelus sp) between Marine Finfish Production And Research Centre (MAFPREC), Tanjung Demong, 22200 Besut, Terengganu, Malaysia and National Fisheries Research and Development Institute (NFRDI), Korea is proposed following decision made at meeting of Malaysia-Korea Collaborative Research Plan in Putrajaya, Kuala Lumpur on the 30 th June, 2011. (does not take place)

CHRONOLOGY COLLABORATIVE R & D• MoU – R & D collaborative with SW Tech Sdn was signed in 2011.– Title: Proposal For Research & Development On Grouper’s Fries Production

– Did not take place

• MoU - R & D collaborative with TADC was signed on Dec, 2012– Title: Projek Kerjasama Penyelidikan Pembangunan Induk Dan Pembenihan Ikan Kerapu (R & D Collaborative on Development of Broodstock and Seed production of Grouper)

– on-going

RESEARCH ON GIANT GROUPER2004 till present

PROGRESS IN RESEARCH ON GIANT GROUPER

• 2004 – 2007 : Acquisition of 120 pieces Giant grouper broodstock (BW 18 kg – 104 kg) from Penang and Langkawi for broodstock development program

• 2005 : Importation of 2.5 cm giant grouper fry & reared as potential broodstock

• 2008 - 2009 : Sex reversal project for 6 month with collaborator Winro Sdn Bhd (incubator) and assisted by an expert from Sun Yet Sen University, Taiwan; implantation technique using MT (Methyl Testosterone) hormone at 3 mg/kg & also using hormone androgen to produce male giant grouper ; ratio 1 male : 2 female

• 2009 : Sex reversal to produce female giant grouper using hormone Estradiol (E2)

PROGRESS IN RESEARCH ON GIANT GROUPER• 2006 - 2010: Induced spawning trial using various hormones; success in induction of ovulation & semen production using HCG but were unsuccessful in larval production s as the eggs did not fertilize.

• 2011 : Experiment on use of Vtg can be used as biomarker indicator for sex determination. Based on quantification of vitellogenin (Vtg) in blood, determining the reproductive status and degree of sexual maturation and can be assessed according to the levels of vitellogenin in plasma.

PROGRESS IN RESEARCH ON GIANT GROUPER

• 2011 & 2012: Hybridization of Tiger Grouper and Giant Grouper

• Mass production of hybrid grouper

• Study on growth performance of hybrid grouper (Epinephelus fuscoguttatus x ♀ E. lanceolatus ) and pure ♂strain tiger grouper (E. fuscoguttatus ) at nursery

EGG DEVELOPMENT OF HYBRID GROUPER (E. fuscoguttatus x E. lanceolatus )

FRI TANJUNG DEMONG,BESUT, TERENGGANU

Fertilized egg 00:00

1 cell stage 00:33

2 cell stage 00:16

4 cell stage 00:11

8 cell stage 00:14

16 cell stage 00:12

32 cell stage 00:19

64 cell stage 00:17

128 cell stage 00:27

Blastula 00:24

Gastrula 01:45

Neurola 02:09

Head & Tail bud 01 : 08

Hatched Larvae 10:09

Total : 18:33Hybrid & Pure

Strain

Pemilihan Induk•Matang dan tiada kecacatan•Saiz oocyte melebihi 400µm

Pengambilan Sperma•Sperma yang diambil terus dari induk•Disimpan dalam krioawetan

Lurutan Telur•Lurutan setelah suntikan hormon•Dibuat dalam bekas yang kering

Persenyawaan- Kacau percampuran sperma & telur- Masukan air laut bersihMembilas Lebihan Sperma- Mengunakan penapis yang sesuai- Masukkan dalam tangki penetasan

0 10 20 30 40 50 60 70 80 900

100

200Egg Yolk

Hour After Hatching (h)

Perc

enta

ge

(%)

0 10 20 30 40 50 60 70 80 900.00

1000.002000.003000.00 MG(µm)

Linear (MG(µm))

Hour After Hatching (h)

(µm)

HYBRID METHODOLOGY

EGG DEVELOPMENT

GRAPH : LARVAE AFTER FHATCHING

Abnormal Larva

Normal Larva

BREEDING TECHNOLOGY OF HYBRID GROUPER

(TIGER GROUPER X GIANT GROUPER )♀ ♂

HYBRID GROUPER LARVAE REARING PROTOCOL UNTIL 30-DAH

(2.0 – 2.5 cm)

2 6 10

14

18

22

26

30

Fish oil

Phytolankton ( Nanno chloropsis)

Rotifer size s/ss

Rotifer size L

Love Larva No 1

L/ Larva No 2

Artemia/kopepod

Water exchange 10-30%

Water exchange 30-

50%

Siphon tank

Day after hatchingFRI TANJUNG DEMONG,

22200 BESUT TERENGGANU096956778

PROGRESS IN RESEARCH ON GIANT GROUPER

• 2013:. Cryopreservation of giant grouper semen

SALINITYConcentrat

ion of NaCl, %

Observation

1 10 No movement

2 18 No movement

3 28 Moving

4 36Move

vigorously

5 45

Move fast

but stop

early

6 55 Slow vibrate

7 62 No movement

FS A B C D E F G H I J K L M0

5

10

15

20

25

17

2

17 1718 18 18

11

1

2122

1

18

6

Tittle: Days of Sperm Survival Extender

Extender

Numbers of Day

PROGRESS IN RESEARCH ON GIANT GROUPER

• 2013: Effect of Polychaete on Gonad Development of Giant Grouper

Size of fish: 25kgStocking rate : 3 fish/10 tonTreatment: Freeze-dried polychaete worm in capsuleTreatment dose: 1 , 2 & 3 capsule/day and controlFeeding regime: alternate days or 3x/weekData collected: Gonad status & Fatty acid composition in plasma.

RESULT:• Fish fed 2 & 3 capsule

produce egg faster than control.

• Fatty acid in polychaete contain high arachidonik acid that gave positive effect on the development of gonad

PROPOSED R & D BETWEEN FRI TD & ECO MARINE FARM

STRENGTH FRI TD STRENGTH ECO MARINE FARMFacilities availableMatured male broodstocks readily availableExperience researchers in breeding, larval rearing

Expert in live feed production:( rotifer & algae)Broodstock maturation dietMatured female broodstock readily available

Proposed Scope of Collaboration

• Broodstock Development Program for Giant Grouper– Cryopreservation of semen– Genetic and disease screening– Broodstock maturation diet

• Breeding Technology of Giant Grouper– Sex reversal– Hormone treatment for artificial spawning

– Spontaneous spawning in controlled conditions

– Study on improving egg quality of giant grouper, Epinephelus lanceolatus

THANK YOU