Natural Product Antifoulants (NPAs) Natural Product Antifoulants (NPAs) – Present Scenario & Future Prospects Raveendran TV, VP Limna Mol & PS Parameswaran National Institute of Oceanography, INDIA
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