Natural Product Antifoulants (NPAs)Natural Product Antifoulants (NPAs)– Present Scenario & Future Prospects

Raveendran TV, VP Limna Mol & PS ParameswaranNational Institute of Oceanography, INDIA

What are NPAs?What are NPAs?

Globally accepted and one of the most promisingGlobally accepted and one of the most promisingenvironmentally benign option to control marinebiofouling

They are secondary metabolites secreted by themarine organisms to control fouling on their surfacesmarine organisms to control fouling on their surfaces

Secondary metabolites: compounds that are noty pessential for growth and reproduction, but rather,through interaction with their environment, enhanceprospects of survival of the organismprospects of survival of the organism

Wh organisms prod ce NPAs?Why organisms produce NPAs?

Sessile soft-bodied marine organisms are havinginherent antifouling property, i.e. they producesecondary metabolites as a chemical means ofsecondary metabolites as a chemical means ofwarfare against:

Protection from predationCompetition for spacePrevention of overgrowth/biofoulingCombat poisoningFight infectionsFight infections

1625 – 1st coatingPatent by william

2003 – IMO ban on TBT i t li tiate t by a

Beale

1600 1700 1800 1900 2000 2005

paint application

2008 – IMO ban on TBT based paints

Copper sheathing and heavy metal based coatings – copper coatings with booster biocides

TBT SPC, 1976 patent

Foul release coatings, 1977 patent

Novel environmentallyyAcceptable alternatives

f fTimeline for key antifouling generationsAdopted from Chambers et al.,2006.

Key environmental issues of marine antifouling Coatings containing biocides

Effect on non-target organismsg gThe length of the time the biocides remain in theenvironmentBuild up in the marine food chain – Bioaccumulationand biomagnification

WHY NPAs?WHY NPAs?

The commonly employed, most economic and longlasting antifouling paint ingredient, TBT, is presentlyfacing a total global ban imposed by IMO

Therefore there is an urgent need for finding

g g p y

Therefore, there is an urgent need for finding environmentally benign and long-lasting

alternative for TBT.

Novel antifouling strategies

Self polishing co-polymer coatingsFoul Release Silicone Coatings Ozone and Hydrogen peroxide release curtainsOzone and Hydrogen peroxide release curtainsBacterial antagonistic product coatingsNPAs based coatingsNPAs based coatings

A l till i t f t ikiAmple scope still exists for striking potential NPAs

With more than70% f th th70% of the earthcovered by oceans,marine naturalmarine naturalproducts representonly 10% of naturalonly 10% of naturalproducts describedto date (2007).to date (2007).

Potential sources for bioactive antifouling compounds

BRYOZOANS

SOFT CORALS

SPONGESWith our Vast EEZ Blessed i h i h bi di i h ASCIDIANS

MANGROVES

with rich biodiversity much needs to be explored

SEA WEEDS

BACTERIA (1 5%BACTERIA (1-5% marine microbes so far isolated)

NPA b d A/F F l tiNPAs based A/F Formulations

SEA NINESEA NINE-- 211 PAINT211 PAINT–– California, U.S.A.California, U.S.A.EuniceaEunicea sp. (soft coral)sp. (soft coral)–– IsothiozolonIsothiozolonI hibitI hibit E t hE t h

SEA NINESEA NINE-- 211 PAINT211 PAINT–– California, U.S.A.California, U.S.A.EuniceaEunicea sp. (soft coral)sp. (soft coral)–– IsothiozolonIsothiozolonI hibitI hibit E t hE t hInhibit Inhibit EnteromorphaEnteromorpha

NETSAFE & PEARLSAFE –A/F products from Australian red seaweed Delisea pulchra Furanone Uvsty of New South Wales

Inhibit Inhibit EnteromorphaEnteromorpha

NETSAFE & PEARLSAFE –A/F products from Australian red seaweed Delisea pulchra Furanone Uvsty of New South Walesseaweed Delisea pulchra – Furanone - Uvsty of New South Wales, Australia – tested on nets in aquaculture farms – effective against marine bacteria, epiphytic seaweeds, barnacles, bryozoans and tube worms

seaweed Delisea pulchra – Furanone - Uvsty of New South Wales, Australia – tested on nets in aquaculture farms – effective against marine bacteria, epiphytic seaweeds, barnacles, bryozoans and tube wormstube worms

SHMRC, Tuticorin – Bryozoan, ZoobotryonZoobotryon pellucidumpellucidum ––TBG (TBG (TribromomethylTribromomethyl graminegramine) High activity ) High activity –– against barnacles against barnacles --

tube worms

SHMRC, Tuticorin – Bryozoan, ZoobotryonZoobotryon pellucidumpellucidum ––TBG (TBG (TribromomethylTribromomethyl graminegramine) High activity ) High activity –– against barnacles against barnacles --(( yy gg ) g y) g y ggPractical test Practical test –– Oil storage vesselOil storage vessel-- ongoingongoing

(( yy gg ) g y) g y ggPractical test Practical test –– Oil storage vesselOil storage vessel-- ongoingongoing

Major Bioactive Antifouling Natural ProductsProducts

TerpenoidsSteroidsCarotenoidsPhenolicsFuranonesAlkaloidsAlkaloidsPeptidesLactonesLactones

SPONGES Indian Scenario

Year Place Source A/F Compound

Activity against

Commercial Status

1989 Vishakapatnam

Dysidea herbacea

Herbacin Bryozoans Hydroids Serpulids

--

1998 NIO, Goa Haliclona sp. Crude Green mussel

--

1999 RRL, Bhuvane

-swar

Azorica pfeifferae

Crude Balanus amphitrite

cyprids--

2002 NIO, Goa Psammaplysilla purpurea

Crude Fouling Bacteria

--

SPONGES World Scenario

Year Place Source A/F Compound

Activity against

Commercial Statusp g

1981 California, U.S.A

Haliclona rubens, H.

viridis

Isothiozolone Seaweeds Sea Nine 211 paint

viridis

1996 Fusetani Biofouling

P j t

Acanthella cavernosa

Terpenoids Balanus amphitrite

id--

Project (FBP), Japan

cyprids

1996 FBP J P d P d ti B l1996 FBP, Japan Pseudoceratina

purpurea

Pseudoceratidine

Balanus amphitrite

cyprids

--

1998 FBP, Japan Acanthella cavernosa

Steroid peroxidase

Mussel, B.a.cyprid

--

SPONGES World Scenario

Year Place Source A/F Compound

Activity against

Commercial Status

1998 FBP, Japan Axinyssa. sp.

Sesquiterpene

B.a. cyprid

--

2001 France Ircinia oros Phenolic Fouling --derivatives

gBacteria

& Diatoms

2002 U.S.A. Verongid sponge sp.

Bromotyrosi-n derivative

B.a. cyprid

--

2003 Italy Reniera APS (Alkyl B a --2003 Italy Reniera sarai

APS (Alkyl Pyridinium

salts)

B.a. cyprid

2004 FBP Japan Acanthella Isocyanide B a --2004 FBP, Japan Acanthella cavernosa

Isocyanide B.a. cyprid

--

SOFT CORALS Indian Scenario

Year Place Source A/F Compound

Activity against

Commercial StatusCompound against

1989 Tuticorin, India

Juncellajuncea,

Crude B. amphitrite

id

--

cyprid1998 NIO, Goa Cladiella

pachyclados, Crude Mussels --

Sinularianumerosa,

1999 NIO, Goa Sinularia Crude Fouling --compressa,

S. leptocladosBacteria

2001 RRL Echinogorgia Crude Fouling2001 RRL, Bhuvane-swar, India

Echinogorgiacomplexa

Crude Fouling diatoms

--

SOFT CORALS World Scenario

Year Place Source A/F Compound

Activity against

Commercial Status

1983 California Leptogorgia virgulata, L.

setacea

Homarine Fouling diatom

1985 U.S.A. Muricea californica, M.

fruticosa

Muricin Fouling diatom

1986 U.S.A. Renilla reniformis Renillafoulins

B.a. cyprid

1987 North Leptogorgia Diterpenoid B a cypri1987 North Calorina

Leptogorgia virgulata

Diterpenoid hydrocarbo

ns

B.a.cyprid, fouling diatoms

1988 G i P d t F F li1988 Georgia Pseudopterogorgia americana

Furanogermacrene

Fouling diatom

SOFT CORALS World Scenario

Y Pl S A/F C d A ti it CommercialYear Place Source A/F Compound Activity against

Commercial Status

1988 U.S.A. Leptogorgia Pukalide, B. a. --virgulata Epoxypukalide cyprid

1993 Japan Sinularia sp. β-bisabolene Mussel --

S1994 Japan Sinularia sp. 13 α-acetoxypukalide

B.a. cyprid

--

2002 Brazil Phyllogorgia Crude Barnacle --dilatata cyprid

Indian Scenario – no work on NPAs from Indian Seaweeds ?

SEAWEEDS World ScenarioYear Place Source A/F Compound Activity against CommercialYear Place Source A/F Compound Activity against Commercial

Status

1994 Brazil Dictyota menstrualis

Dictyols (Diterpenes)

Bryozoan --

1995 Australia Delisea pulchra

Furanones B. a. cyprids, seaweed

--

2002 Brazil Laurencia bt

Elatol Mussel --abtusa

2002 Brazil L. rigida Sesquiterpenes, elatol,

Dischloroelatol

Invertebrate larvae, Algal

spores

--

2002 UK Zostera marina

Zosteric acid Barnacle cyprid, bacteria, algae,

tubeworms2004 F Bif i Li dit B l id2004 France Bifurcaria

bifurcataLinear diterpenes Barnacle cyprids,

fouling bacteria--

ASCIDIANS Indian Scenario

Year Place Source A/F Compound

Activity against

Commercial Status

1989 Tuticorin Styela Crude Artemia1989 Tuticorin, India

Styelapigmentata,Pyura pallida

Crude Artemiasalina &

microalgae

--

ASCIDIANS World Scenario

Year Place Source A/F Compound

Activity against

Commercial Status

1991 France Polysyncraton lacazei

Fractions Sea Urchin larvae

--

1994 UK Clavelina lepadiformis

Crude Invertebrates --

1999 Morocco Cynthia savignyi

Crude Artemia salina

--

2003 UK Amaroucium stellatum

Crude Balanusamphitrite

--stellatum, Botryllus

planus

amphitrite cyprids

Natural Products Antifouling LaboratoryNatural Products Antifouling Laboratory National Institute of Oceanography,

Regional Centre, Kochi

Initiated research on NPAs from 2004Initiated research on NPAs from 2004

Set up Basic infrastructure for conducting A/Ft titesting

Developed manpower & expertise in the variousDeveloped manpower & expertise in the variousantifouling assay techniques and the isolation &characterization of NPAs

Field trips undertakenField trips undertaken

Conducted a few shallow coastal water as well as opensea SCUBA diving operations for sample collection

Sampling locations:Rameswaram &L k h d I l dLakshadweep Islands

Screened 36 species of sponges29 species of soft corals

Test organisms for antifouling assayg g y

Diatoms Amphora coffeaeformisCyprids of barnacles Balanus amphitrite

Bacteria Fouling bacterial strains

NPAs detected during the current study

6- Hydroxy polyanthellin, a novel tetracyclic diterpenoid

(Cladiella sp.)

Araguspongine C(Acanthella cavernosa) F i A id(Acanthella cavernosa) Furoic Acid,

sesquiterpene (Sinularia kavarattiensis)

A possible research management schedule for the development of a novelantifouling system based on natural antifouling compounds, Yebra et al 2004

YEAR

Hypothesis 0 12 3 4 5 6 7 8 9Collection and extraction of organisms

YEAR

Collection and extraction of organisms

Antifouling screening test/fractionation

Identification of antifouling compounds

Patent registration

Small scale synthesis

Antifouling tests: bioassays, field testsg y

Ecotoxicological testing

Commercial assessmentIndustrial synthesis

The effective concentrations of various natural products to inhibit 50% of the tested population of the barnacle Balanus amphitrite

10.275

0.25

0.3µg

ml -

1

0.175

0.15

0.2

EC50

/ µ

0.075

0.02

0.07

0

0.05

0.1

E 0

Sponge (Acanthella cavernosa) Sponge (Reneira sarai)Algae (Delisea pulchra) Mollusca (Phyllidia pustulosa)

Natural product source

Algae (Delisea pulchra) Mollusca (Phyllidia pustulosa)Tunicate (Eudistoma sp.)

Natural product sourceThe halogenated furanones from the red seaweed Delisea pulchra has proved the most successful, with EC50 being as low as 0.02 µg ml -1

Phyletic distribution of 160 reviewed marine species from which

Of the 160 potential NPAs, 76% are from sponges, algae & cnidarians

Phyletic distribution of 160 reviewed marine species from whichpotential antifouling natural products have been extracted(Adopted from Chambers et al., 2006)

NPAs

Advantages

s

AdvantagesVery specific in actionBiodegradable compoundE f i dl i tEco-friendly in nature

DisadvantagesDisadvantagesVery poor yieldStructure elucidation

Requirements for an optimal antifouling coatingAdopted from Chambers, 2006

Must be Must not be

AnticorrosiveAntifoulingEnvironmentally acceptableEconomically viable

Toxic to the environmentPersistent in the environmentExpensiveChemically unstableEconomically viable

Long lifeCompatible with the underlying systemResistant to abrasion/biodegradation/erosionCapable of protecting regardless of operational profile

Chemically unstableA target for non-specific species

Capable of protecting regardless of operational profileSmooth

“FOR EVERY NATURAL CAUSEFOR EVERY NATURAL CAUSE THERE IS A NATURAL SOLUTION”

LET US HAVE THE PATIENCE

TO IDENTIFY AND IMPLEMENT IT

TO PROTECT & PRESERVE MOTHER NATURETO PROTECT & PRESERVE MOTHER NATURE

FOR THE FUTURE GENERATIONS

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