SEMINAR TRANDISIPLINARI DALAM PENYELIDIKAN

KLINIKAL

16. 12. 2008

Associate Prof. Dr. Farid Che GhazaliSenior LecturerSchool of Health Sciences, USM, 16150 Kubang Kerian, Kelantan.Tel: 09 766 3833 H/P: 019 910 9642 Fax: 09 764 7884Email: farid@kb.usm.my

En Kamarruddin IbrahimTimbalan Ketua PengarahJabatan Taman laut MalaysiaPUTRAJAYA

Mr Panner SSenior Lecturer in Forensic SciencesSchool of Health Sciences, USM, 16150 Kubang Kerian, Kelantan.Tel: 09 766 3833

Associate Professor. Dr. K.N.S.SirajudeenDepartment of Chemical Pathology, School of Medical Sciences, USM, USM, 16150 Kubang Kerian, Kelantan

Crown-of-thorns (COT) biology and measures of sustaining its outbreak and elucidation of potential

therapeutic - potential neutraceuticals.

Crown-of-thorns starfish

                                     

Scientific Classification

• Kingdom: Animalia • Phylum: Echinodermata • Class: Astoridea • Order: Valvatida • Family: Acanthasteridae • Genus: Acanthaster • Species: A. planci

Binomial name

Acanthaster planci

The crown-of-thorns starfish (Acanthaster planci) is a member of the Phylum Echinodermata, which also includes the Sea Urchin, Sea Cucumber, and Sand Dollar. The crown-of-thorns can have up to 20 arms extending from the central disk and has thorn-like spines all over its body. Acanthasters are carnivorous. They everts their stomach outside the body over "poor, defenseless" living coral polyps. This is one of the few sea animals that feed off on living coral.

Cots' should be handled carefully because they have pointy spines (which release a neurotoxin) and are venomous. If poked the wound will eventually turn blue and will sting and swells that may last for a couple of days.

• Outbreaks of crown-of-thorns starfish have been a major issue on the Great Barrier Reef and other Indo-Pacific reefs for nearly 40 years. The outbreaks have generated great concern among the community and considerable debate among scientists. Outbreaks generally occur at regular intervals with coral cover returning to pre-outbreak levels in the intervening years.

• Although the issue is globally acknowledged and is purported to be affecting the marine coral areas of Malaysia there is lack of documentation and reports available. Action and guidelines should be established before disastrous outbreak as reported in 2003 that affects reefs between Cairns and the Whitsundays which costed tourism operators, and the Queensland and Commonwealth Governments about $3 million a year for control measures till date.

Problem Statement

There is no comprehensive documentation on the causes of outbreaks of these star fish and their effect on coral reef in Malaysian marine ecosystem. There is a ministerial question whether the neurotoxin produced is of therapeutic potential ? Hence a comprehensive multifaceted research study is proposed on crown of thorn starfish.

This includes:

Electron microscopy and morphological studiesSubstrate utilization and biochemical characteristicsChemotaxonomic characterizationGene sequencing and phylogenetic analysisToxin productionCloning and sequencing of amplified product

1. To understand the distribution of crown-of-thorns starfish in Malaysia marine, coastal and reef areas.

2. To achieve a national database that characterize better understanding of the biology of the starfish involving microscopical and molecular works especially COT that is identified within the Malaysian marine and coastal areas.

3. To elucidate and features the starfish inert chemical characterization of therapeutic potential.

4. To record the history and nature of the outbreaks and the pattern of recovery pertaining to Malaysian marine and coastal reefs.

What is the general objective?

Objectives of this study:

1. To analyze the total lipid content, lipid class composition and fatty acid composition of starfish Acanthus Planci.

2. To determine the effect of starfish Acanthus Planci Phosphatidyl choline (PC) on the cultured colon cancer cell line.

3. To determine the antioxidant capacity of Starfish by FRAP assay.

Analysis of Lipids on the Star fish (Acanthus Planci)

Molecular characterizations of Acanthus Planci

Objectives of this study:

1. To identify the mtDNA based molecular genetic marker for Acanthus planci in Malaysian marine waters by amplifying and sequencing COI and 16SRNA

2. To compare the sequence with the sequence reported in the literature.

3. This will help to identify the Acanthus planci larvae so that effective control measure can be taken before the maturity of the coral destroyer

Antioxidant activity of starfish homogenates by FRAP assay

Starfish collecti

on, storage

Total lipid extraction from the starfish homogenates

Separation of polar and non polar lipids, lipid composition analysis and

fatty acid analysis

Separation of Phosphatidyl choline (PC)

Caco-2 cell culture and effect of PC• Cell viability (assay) and

microscopic assay

DATA ANALYSIS and REPORT SUBMISSION

Expected outcome:

Human Resource Development - one research officer (RA) will be trained.

Publication/Presentation - one or two

Multi-disciplinary Knowledge development and applications:

1. To better understand the lipid composition of starfish Acanthus Planci and its effect on cancer cells.

2. If it shows the promising results, then it could be utilized for the commercial production of EPA from this starfish and helpful in the development of anticancer agent.

Analysis of Lipids on the Star fish (Acanthus Planci)

Expected outcome:

Human Resource Development - one research officer (RA) will be trained.

Publication/Presentation - one or two

Knowledge development and application:

Development of molecular marker to identify Acanthus planci and their larvae to effectively monitor the outbreak and take preventive steps

Duration of the research project 3 years

Total Expected expenditure : RM 150,000-175,000

Equipments required for the study:Gas ChromatographELISA readerFluorescence MicroscopeSpectrophotometer

Total Expected expenditure RM 190,000

Dedicated PCR machine - RM 30,000Liquid nitrogen chamber small and big - RM 10,000Kits and disposables - RM 60,000Sequencing using external facility - RM30,000Salary for RA - RM 60,000

The outcome of the projects:

• Guideline and actionline (of control) of COT outbreak and preservation of marine park ecology.

• Therapeutic nutriceuticals usefulness from COT

Expected outcome:

Human Resource Development - one research officer (RA) will be trained.

Publication/Presentation - one or two

Knowledge development and application:

Development of molecular marker to identify Acanthus planci and their larvae to effectively monitor the outbreak and take preventive steps.

SEKIAN TERIMA KASIH