Name : Muhammad Farouk bin Harman Matric No. : UK20532 S.M.S.G.

(Fishery) Supervisor : Dr Ikhwanuddin

Fertilization procedure. Sperms are directly placed into the reproductive tract of a female by using means other than sexual intercourse or natural insemination (Batul 2011)

Mud spiny lobster, Panulirus polyphagus Belongs to Family Palinuroidae Live on muddy substrates and sometimes on rocky bottoms. Depth range from 3 to 90 meter, usually far less than 40 m.
(Herbst, 1793)

Panulirus polyphagus

Collected widely by human High commercial value. Big deacreased in landings.

1103tonnes in 2000 and 274tonnes in 2009 (FAO 2000-2009)

Farmers taking wild seeds

Preventing recruitment for juveniles of P.polyphagus .
4

The female can be distinguish from the male by its swimming legs where it is bigger and have double pereiopods for holding the fertilized eggs.

Double pereiopods

Figure 1a: Female mud spiny lobster

The male can be distinguish from the female by its swimming legs where it is smaller and have single pereipod.

Single pereiopod

Figure 1b: Male mud spiny lobster.

Lobsters are invertebrates, which belong to the Crustacean class of the Phylum Arthropoda.(Velantino, 2010) In many parts of the world, lobsters support commercially valuable fisheries, in regions the most economically important one.
(FAO, 2004)

Landing(Tonnes)
2500

Total Landing of Mud Spiny Lobster Through 2000-2009 in Malaysia

2059 2000 1700

1612 1500

1103 1000

1143

Total Landings Each Year

500 247

433 200 255 274

0 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009

Year

The lobster has always been a favorite of the general public and in early 1970s this popularity was utilized to promote the idea of the lobster aquaculture. (Aiken and Waddy, 1995) The culture of spiny lobsters in Malaysia is still in its infancy, with activities based in the State of Sabah. (Tuan and Mao, 2004)

At present, lobster seed is sourced from the wild.

(Tuan and Mao, 2004)

Pueruli (juveniles of Panulirus polyphagus) are not yet commercially from hatcheries and so must come from the wild. (Booth and Phillips, 1994) Spiny lobster has a complex larval development, with many developmental stages making it extremely difficult to raise seeds for aquaculture in a hatchery.(Booth and Phillips, 1994)

1)

The seed production technology of lobsters, Panulirus polyphagus has not yet been develop.
A hatchery production of spiny lobster seed is not yet developed. Even if successful, seed production will be very Pitcher, 1993) difficult technically and prolonged, making it high risk.
(Adams

and Dalzell, 1993; Linton, 1998; Munro, 2000;).

2) Difficulty for getting the mature male of mud spiny

lobster, Panulirus polyphagus.

Size at onset of maturity is more difficult to assess in male than in female lobsters. Male physiological maturity has been determined by the presence of mature spermatozoa in vasa deferentia of several spiny lobsters.
(Heydorn, 1969a; Berry, 1970; MacDiarmid, 1989a; Turner et al., 2002)

Artificial insemination could improve the efficiency of fertilization rate and thus predictability of seed production.
(Bart et al., 2006)

Artificial insemination remains as one of the most important assisted reproductive technologies. (Vishwanath, 2003)

The remarkable progress made in other assisted reproductive technologies does have the potential to rapidly generate offspring.(Vishwanath, 2003)

There are no availability of a perfected hatchery technology and limited availability of juveniles and subadults from the wild. (Radhakrishnan, 1996)

To develop artificial insemination technique through induced molting on mud spiny lobster Panulirus polyphagus. To determine the success of fecundity and hatching rate using artificial insemination technique on mud spiny lobster, Panulirus polyphagus.

The study will be carry out at the Marine Hatchery of the Universiti Malaysia Terengganu (UMT) and scheduled to be conducted from January 2012 until April 2012. 3,000 Liter capacity tank is prepared for 5 female, 5 male and a pair of female and male as a control. Forceps for taking out the spermatophore from male and inserting it into the soft thelycum of female.

Sampling and rearing the mud spiny lobster, Panulirus polyphagus

Induced molting

Extracting spermatophore

Artificial insemination technique

Data analysis
Figure 3: Steps in the experiment.

METHODOLOGY
Experimental design

The species in this study is mud spiny lobster, Panulirus polyphagus.

For the control, a pair of female and male of mud spiny lobsters, Panulirus polyphagus will be rear as pairs (1 male and 1 female) in a tank filled with 80% filtered seawater maintained with constant gentle aeration. For the project purposes, one tank will be rear with 5 female and one tank will be rear with 5 male. Female tank salinity will be manipulated in order to induced the molting of the female mud spiny lobster. The mud spiny lobsters will be fed on squid flesh.

12 spawners Induced Molting (10 spawners)

Molted spawners (10 spawaners) Artificial Insemination

Natural spawners (2 spawners)

Fecundity calculation

Success of hatching (survival rate)

Compare of fecundity and hatching rate from one artificial insemination female and one control female(natural spawner).
Figure 4: Experimental Design

Induced Molting
Salinity manipulating technique

Salinity of 2ppt will be decrease from 30 ppt each day until 24 ppt and increase 2ppt until 30 ppt.(repeated until molt)

Female that ready to molt will be seperate from the others.

Once the female molt, it is prepared for artificial insemination technique.

Artificial Insemination Steps

Spermatophore from the mature male lobster are taken between the 4th and 5th walking legs using a forcep.

The spermatophore will be insert into the soft thelycum of female that recently went through moulting.

Temperature of 24C-26C. Salinity of 24-30ppt. Ammonia level: <0.025 ppm. Dissolved oxygen: > 5 ppm pH : 6.5-8.5

YSI meter model 85. Ammonia test will be run at water quality lab.

1)Fecundity = no. of eggs/female = no. of larvae hatched + no. of none hatched eggs 2) Hatching Rate = Number of hatched larvae X 100 Fecundity

These data will then be process using the Excel and SPSS software using the One-way ANOVA test.

Artificial insemination become valuable in aquaculture industry of mud spiny lobster. Increase the hatching and survival rate of the larvae.

Year Activities Preparation, proposal writing and presentation. Sampling and rearing the mud spiny lobster, Panulirus polyphagus Induced molting. Artificial insemination technique Statistical analysis Excel & SPSS. Thesis writing, final presentation and submission.

2011 September October November December January February

2012 March April May

Booth, J. D. and Phillips, B. F. 1994. "Early Life History of Spiny Lobster." Crustaceana 66(3): 271-294. Tuan, L.A. and Mao, N.D., 2004. Present status of lobster cage culture in Vietnam. Proceeding of the ACIAR lobster ecology workshop, p24-30. Institute of Oceanography, Nha Trang, Vietnam. Vishwanath, R. 2003. Artificial insemination: the state of the art. Theriogenology 59, 571-584. Radhakrishnan, E. V. 1996. Lobster farming in India. 48 : 96 98. Batul N.B. Artificial insemination (http://www.buzzle.com/articles/how-is-artificial-insemination-done.html) [Access on 17/11/2011] Velantino, S. 2010. Lobster biology. (http://ezinearticles.com/?LobsterBiology&id=408797) [Access on 16/11/2011]
Balss, H. 1914. Marine lobsters Of The World (http://speciesidentification.org/species.php?species_group=lobsters&menuentry=soorten&id=162&tab=ref s) [Access on 17/11/2011]

5155 mm Carapace Length for males. 5160 mm Carapace Length for females. (Kizhakudan and Patel)

Area of spermatophore will be taken.

Adult

Juvenile

Early phyllosoma

Puerulus

Late phyllosoma

Eggs and fertilized eggs is left with its mother until hatching.

Booth, J. D. and Phillips, B. F. 1994. "Early Life History of Spiny Lobster." Crustaceana 66(3): 271-294. Synthesize current information on the lobster early life history Discovery of the habitat young juveniles lead to examination of relationship peruli abundance at sea, level of settlement, abundance of juveniles and recruitment. Tuan, L.A. and Mao, N.D., 2004. Present status of lobster cage culture in Vietnam. Proceeding of the ACIAR lobster ecology workshop, p24-30. Institute of Oceanography, Nha Trang, Vietnam. Reviews current status of lobster cage culture, identify major technical and sosio-economic constraint to further development. Vishwanath, R. 2003. Artificial insemination: the state of the art. Theriogenology 59, 571-584. Radhakrishnan, E. V. 1996. Lobster farming in India. 48 : 96 98. The potential for increasing the yield from the wild fisheries is limited, as the stock is subjected to extreme fishing pressure and is nearing their long term equilibrium yield. Hence, the most imaginative management strategy seems to be increasing the production through population enhancement and aquaculture

Batul N.B. Artificial insemination (http://www.buzzle.com/articles/how-is-artificial-insemination-done.html) Explaining on Artificial Insemination [Access on 17/11/2011] Velantino, S. 2010. Lobster biology. () [Access on 16/11/2011] http://ezinearticles.com/?Lobster-Biology&id=408797 Explain on lobster morphology
Balss, H. 1914. Marine lobsters Of The World (http://speciesidentification.org/species.php?species_group=lobsters&menuentry=soorten&id=162&tab=refs) [Access on 17/11/2011] Describe on general things on Panulirus polyphagus

Booth, J. D. and Phillips, B. F. 1994. "Early Life History of Spiny Lobster." Crustaceana 66(3): 271-294. FAO, 2004. Fishery statistics. Capture production 2002. FAO Yearbook. 94/1. FAO, Rome. Aiken, D. E and Waddy, S. L. 1995. Chapter 8 : Aquaculture. Journal of Biology.153-175 Pratoomchat, B. , Piyatiratitivorakul, S. and Menasveta, P. 1993. Sperm quality of pond reared and wild-caught Penaeus monodon in Thailand. Journal of the World Mariculture Society 24: 530-540. Velantino, S. 2010. Lobster biology. (http://ezinearticles.com/?LobsterBiology&id=408797) [Access on 16/11/2011]

Balss, H. 1914. Marine lobsters Of The World (http://speciesidentification.org/species.php?species_group=lobsters&menuentry=soo rten&id=162&tab=refs) [Access on 17/11/2011]

Tuan, L.A. and Mao, N.D., 2004. Present status of lobster cage culture in Vietnam. Proceeding of the ACIAR lobster ecology workshop, p24-30. Institute of Oceanography, Nha Trang, Vietnam. Petersen, E. H. and Phuong, T. H. 2010. "Tropical spiny lobster (Panulirus ornatus) farming in Vietnam bioeconomics and perceived constraints to development." Aquaculture Research Adams, T. and Dalzell, P. 1993. Pacific Island lobster fisheries: Bonanza or bankruptcy? Fisheries Newsletter #67, South Pacific Commission, New Caledonia. 9. Vishwanath, R. 2003. Artificial insemination: the state of the art. Theriogenology 59, 571-584. Gwo, J.C. 2000. Cryopreservation of aquatic invertebrate semen: a review. Aquaculture Research, 31, 259-271 Radhakrishnan, E. V. 1996. Lobster farming in India. 48 : 96 98. Bart, A. N., Choosuk, S. and Thakur, D. P. 2006. Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fibricius). Aquaculture Research, 37,523-528. Batul N.B. Artificial insemination (http://www.buzzle.com/articles/how-is-artificial-insemination-done.html) [Access on 17/11/2011]