EnvironmentAsia

The international journal published by the Thai Society of Higher Education Institutes on Environment
Available online at www.tshe.org/ea/index.html
EnvironmentAsia
Available
DOI
9(2) (2016) 48-54
online at www.tshe.org/EA
EnvironmentAsia 2 (2009) 50-54

Genotoxicity Assessment of Mercuric Chloride in the Marine Fish Therapon jaruba

Qualitative Assessment and Management of Microplastics in
NagarajanAsian
Nagarani,
GreenArumugam
Mussels Kuppusamy Kumaraguru,
(Perna viridis) CulturedVelmurugan Janaki Devi
in Bacoor Bay,
and Chandrasekaran Archana Devi
Cavite, Phillipines
Center for Marine and Coastal
Cristian Studies,
Ryan School
Argamino andofJose
Energy, Environment
Isagani B. Janairoand Natural Resources,
Madurai Kamaraj University, Madurai-625021, India
Biology Department, De La Salle University, 2401 Taft Avenue, Manila 0922 Philippines
Abstract

The aim of the present study was to standardize and to assess the predictive value of the cytogenetic analysis
Abstract
by Micronucleus (MN) test in fish erythrocytes as a biomarker for marine environmental contamination. Micronucleus
frequency
baseline(>
Microplastics in5erythrocytes was evaluated
mm) have gained popularityininand genotoxic
research potential
and the public of
eyea common chemical
in recent years. Thiswas determined
is due to the fact
in
that they contain persistent organic pollutants (POPs) which pose potential risks to the environment and humanfor
fish experimentally exposed in aquarium under controlled conditions. Fish (Therapon jaruba) were exposed 96
health.
hrs to a single heavy metal (mercuric chloride). Chromosomal damage was determined as micronuclei frequency
Bivalves, which are filter feeders, are considered to be good indicators of marine pollution. In this preliminary study, Asian in
fish erythrocytes. Significant increase in MN frequency was observed in erythrocytes of fish exposed to
green mussel (Perna viridis), an example of edible bivalve, cultured in Bacoor Bay, Cavite, Philippines was subjected tomercuric
chloride.
qualitativeConcentration of 0.25 ppm
analysis to determine the induced
presencethe
ofhighest MN frequency
microplastics. Through(2.95 micronucleated
microscopic cells/1000
analysis, cells compared
microplastics were found
to
present in the acid-digested mussel soft tissue. A management program is suggested for policy makers andofstakeholders
1 MNcell/1000 cells in control animals). The study revealed that micronucleus test, as an index cumulativeto
exposure,
reduce the appears
negativeto be a sensitive
impact modelpollution
of microplastic to evaluate genotoxic
to both humanscompounds in fish
and the marine under controlled conditions.
environment.
Keywords:
Keywords: genotoxicity;
microplastics;mercuric chloride;
qualitative micronucleus
assessment; Perna viridis; marine pollution; environmental management

1. Introduction
1. Introduction laboratory
MMT (Million and field conditions.
Metric Tons) perIn 2006
year Soumendra
(Jambeck et al.,
2015). Marine plastic debris primarilybiomarkers
et al., made an attempt to detect genetic come from
In India,are
Plastics about
popular 200due tons of mercury
to several and such
properties its in two fish sources
land-based species,(80%) bata and
Labeothrough Oreochromis
leakage while only
compounds are introduced into the environment
as weight, strength and cost. The rise of consumption mossambica, by MN and binucleate
marine aquaculture and fisheries only contribute (BN)
20%
annually as effluents
over the past few decadesfromhas industries
been a (Saffi, 1981).
huge challenge erythrocytes in the gill and kidney
of these pollutants (Ocean Conservancy, 2015). In erythrocytes
Mercuric
in terms of chloride has been
controlling used in agriculture
environmental pollution.asThe a exposed
recent years, to the
thermal power
emergence of aplant discharge
form plastic at
has been
fungicide, in medicine as a topical
increase in the number of plastics amassed in the antiseptic and Titagarh Thermal Power Plant, Kolkata, India.
studied for its role in pollution - microplastics. Although
disinfectant,
environment and in chemistry
is mainly causedasby an their
intermediate
inert prop- in thereThe haspresent
been no study was conducted
universally accepted to definition
determineat
the
ertyproduction of other mercury
(slow degradation rate) andcompounds.
usage (improper The the
the acute
moment genotoxicity of the heavy
(Van Cauwenberghe et al.,metal compound
2015), individual
contamination of aquatic ecosystems
disposal of plastic waste) (Nor and Obbard, 2014). by heavy HgCl
and group
2
in static systems. Mercuric chloride
researches have contributed their is toxic,
own
metals
Despiteand pesticides
their benefits,hasthe gained increasing
contribution of attention
plastic to solvable in water hence it can penetrate
description of microplastics. In 2004, microplastic the aquatic
in recent decades.
environmental Chronic
degradation has exposure to andin
been significant animals.
size wasMutagenic
defined at studies
aroundwith native fish
20 microns species
(Thompson
accumulation of these chemicals in aquatic
causing damage to marine organisms. Previous studies biota represent
et al., 2004).anInimportant effort
2009, the size in determining
adapted was <5 mm. the This
can result in tissue burdens that produce
have shown that plastics (i.e. microplastics) are ingestedadverse potential effects of toxic agents.
characterization of microplastic size was also This study was
accepted
effects not only
at different trophicin the directly
levels whichexposed organisms,
can potentially cause carried out to
by the Joint evaluate
Group the use
of Experts onofthethe micronucleus
Scientific Aspects
but also in human beings.
bioaccumulation. This phenomenon can affect test (MN) for the estimation of aquatic
of Marine Environmental Protection (GESAMP) (Nor pollution
Fish provides
biological processesa of suitable
marinemodel for monitoring
organisms. In addition, using marine 2014).
and Obbard, edible Other
fish under lab conditions.
researchers have presented
aquatic
food safety and human health are also at risk,quality
genotoxicity and wastewater if these microplastics as particles with <1 mm size which
because of its ability to metabolize
affected organisms are consumed (Vandermeersch xenobiotics and 2. Materials
is argued to beand
moremethods
instinctive since micro refers
accumulated
et al., 2015). pollutants. A micronucleus assay has to the micrometer range (Van Cauwenberghe et al.,
been
used successfully in several species (De
It has been estimated that up to about 12.7 million Flora, 2.1.
2015).Sample Collection
As a marine pollutant, microplastics exist in two
et al.,of1993,
tons plasticAl-Sabti
still ended andup Metcalfe, 1995).
in the ocean in The
2010 forms - primary and secondary. Primary microplas-
micronucleus (MN) test has been
(Jambeck et al., 2015; Andrady and Neal, 2009; Van developed tics,The fish species
directly move intoselected
water for the through
bodies present study
ground
together
Cauwenberghe withet DNA-unwinding
al., 2015). The Philippines assays was as was collected from Pudhumadam coast
runoff. These are mainly composed of virgin of Gulf of
plastic
perspective methods for mass
ranked No. 3 by mass of mismanaged plastic wastemonitoring of Mannar, Southeast Coast of India. Therapon
pellets, scrubbers, and microbeads. These plastic forms
clastogenicity
following China and and
genotoxicity
Indonesia. in fish and mussels
In 2010, plastic jarbua belongs
are abundant to the order
in cosmetic Perciformes
products of theas
like exfoliants
(Dailianis et al., 2003).
marine debris in the Philippines ranged from 0.28-0.75 family Theraponidae. The fish species, Therapon
well as industrial abrasives (Andrady, 2011). On the
The MN tests have been successfully used as jarbua (6-6.3 cm in length and 4-4.25 g in weight)
a measure of genotoxic stress in fish, under both was selected for the detection of genotoxic effect
 C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54

other hand, secondary microplastics are degradation
products of larger plastics (mesoplastic/macroplastic)
which are already in the ocean or seashore. Microplastics
in this form are produced via several pathways such
as mechanical, photo (oxidative) and/ or biological
degradation (Masura et al., 2015). Majority of
microplastics in the marine environment are
secondary microplastics formed through the weathering
of plastic in the seashore. Plastics on beaches have a
faster degradation rate due to the higher temperature
(of sand) as compared to both plastics deep in the
ocean or floating on the water surface (Andrady, 2011).
Mussels, locally known as tahong, are generally
defined as mollusk bivalves which grow in the wild
or through aquaculture. These suspension feeders are
considered to be one of the best biological indicators
of marine pollution because of various characteristics
including their geographical distribution. Their tissues
accumulate pollutants due to their feeding mechanism
and their tendency to stay attached to surfaces which
make them relatively inactive in terms of mobility
thus rendering them as good pollution gauges. They
can also be easily sampled and thus, allow for frequent
experiments and monitoring (Vasanthi et al., 2012;
Figueiras et al., 2002; Chase et al., 2001). The species
of mussel used in this experiment is Perna viridis
(P.vidiridis). P. viridis is a large species of mussel
ranging from 8-16 cm (Rajagopal et al., 2006). Their
habitat includes hard surfaces (rocks, pilings and
floating docks, as well as sandy or muddy bottoms about
a foot below the low tide mark) (McGuire and Stevely,
2009). Their diet includes microscopic phytoplankton,
zooplankton and suspended organic detritus in the water
(Rajagopal et al., 2006). In the Philippines, P. viridis
or tahong is considered as one of the seven major
aquaculture species (Food and Agricultural
Organization, 2006). Aquaculture of tahong can
be traced back to 1955 which was initiated by the
Philippine Bureau of Fisheries and Aquatic
Resources in Binakayan, Cavite (Yap, 1999). Al-
though it is a source of food locally, discretion is
advised for consumption due to their tendency for
bioaccumulation of toxins substances detrimental to
human health (McGuire and Stevely, 2009). Thus,
determining the presence of microplastics in these
marine species will not only be instrumental to
profiling marine pollution but will also serve as a tool
in improving guidelines for food safety.
In this study, we present our findings regarding the
presence of microplastics in cultured bivalves available
for human consumption. These particles were positively
identified through microscopic analysis and other
qualitative criteria. In addition, management procedures
according to best practices were also discussed which
will aid policy makers in reducing the risk posed by
these pollutants to the environment and human health.

Figure 1. Map of Bacoor Bay (Google

Figure
Maps)
1. Map of Bacoor Bay (Google Maps)

3. Results and Discussion 3. Results and Discussion

Previous studies confirmed the presence

Previousof
studies
microplastics
confirmedfrom
themarine
presence
samples
of microplastics
using different
from marine samples
methods. Raman Spectroscopy (Vanmethods.
Cauwenberghe
Raman Spectroscopy
49 and Janssen,
(Van
2014),
Cauwenberghe
Fourier Transform
and Janssen,
Infrared
2014), Fourier Tran
Spectroscopy coupled with eitherSpectroscopy
a Microscopecoupled
or an with
Attenuated
either aTotal
Microscope
Reflectance
or anattachment
Attenuated(Nor
Total Reflectance at
and Obbard, 2014), as well as the
andFluorescence
Obbard, 2014),
Microscope
as well as(Noren,
the Fluorescence
2007) wereMicroscope
utilized by (Noren,
various 2007) were utiliz
 a) Noarecellular
a) No cellular or organic structures visibleorinorganic structures
the plastic are visible in the plastic particle/fibre.
particle/fibre.
b) Clear
b) Clear and homogeneously colored and homogeneously
particles (blue, red, black colored particles (blue, red, black and yellow)
and yellow)
c) If the particle transparent orc)whitish,
If the particle transparent
it shall be examinedorwith whitish,
extraitcare
shallinbe examined with extra care i
a microscope
under high magnification. under high magnification.
Figs. 2 and 3 show examples
C. Ryan ofFigs.
Argamino al. /2EnvironmentAsia
and 3 show
microplastics
et examples
observed9(2) in of48-54
each
(2016) microplastics observed in each of the five samp
of the five samples.

Figure 2. Semi-transparent microplastics

Figure 2. Semi-transparent microplastics

2. Materials and Methods 3. Results and Discussion

Bacoor Bay-cultured samples (P.viridis) were
obtained from Sineguelasan Seafood Terminal in
Bacoor, Cavite, Philippines. The mussel samples were
harvested on January 2016. Fig. 1 shows a map of
Metro Manila and its adjacent provinces. The drop pin
marks the location of Bacoor Bay where the samples
were cultured and harvested. All glassware were
thoroughly cleaned and rinsed with filtered deionized
water (Elga PURELAB Flex) to avoid contamination
dilution (Van Cauwenberghe and Janssen, 2014).
Wet digestion using acid was used to extract
the microplastics for the samples (Vandermeersch
et al., 2015). Twenty milliliters (20 mL) of 70% HNO3
(UNIVAR) were added to three mussels in an
Erlenmeyer flask. Five replicates were prepared.
Mussel tissues in acid were left in in the hood for 40
hours to achieve optimum digestion. The samples were
then heated until boiling for 15-20 minutes using a hot
plate to evaporate the acid and dry the sample. Twenty
milliliters of warm deionized water (~80 degrees
Celsius) were added to each vessel for dilution (Van
Cauwenberghe and Janssen, 2014). Digested samples
were then subjected to vacuum filtration using a
Buchner funnel and Whatman Filter Paper Grade 1
(11-micron pore size). Filters were dried for 2 hours
in an oven at 40 degrees Celsius. Dried filters were
analyzed for the presence of microplastics using a Nikon
SMZ 745T Stereomicroscope (Van Cauwenberghe and
Janssen, 2014; Song et al., 2015).
Previous studies confirmed the presence of
microplastics from marine samples using different
methods. Raman Spectroscopy (Van Cauwenberghe
and Janssen, 2014), Fourier Transform Infrared
Spectroscopy coupled with either a Microscope or
an Attenuated Total Reflectance attachment (Nor and
Obbard, 2014), as well as the Fluorescence Microscope
(Noren, 2007) were utilized by various studies. The
absence of these instruments should not be a hindrance
in the analysis of this important class of pollutants.
Thus, the following criteria were used in determining
the presence of microplastics (Noren, 2007):
a) No cellular or organic structures are visible in
the plastic particle/fibre.
b) Clear and homogeneously colored particles
(blue, red, black and yellow)
c) If the particle transparent or whitish, it shall be
examined with extra care in a microscope under high
magnification.
Figs. 2 and 3 show examples of microplastics
observed in each of the five samples.
Microplastics observed in the samples were all
found to be < 1mm. Transparent whitish and reddish
particles measuring at around 10 to 30 microns (0.1
to 0.3 mm) were observed in Samples 1, 2, and 5.
On the other hand, blue fibers (~0.5 mm in length)
were observed in Sample 3 and 4. These observations
qualify in the abovementioned criteria (Noren, 2007)

Figure 3. Fiber micropplastics Figure 3. Fiber micropplastics

Microplasticcs observed ini the sampllesMicroplastic

were all found
f cs observed
to be <ini 1mm.
the sampl
Traansparent
les were whall
hitish
found
f andto be < 1mm. Traan
reddish particles meeasuring at arroundreddish
10 to 30particles
micronsme(0.1
(easuring
to 0.3atmm)
marroundwere
10ob to
bserved
30 microns
in Saamples
(0.1
( to1,0.3 mm)
m were obbse
2, and 5. On the other hand, bluee fibers
2, (~0.5
and 55.mm 50
On in
theleng
oth
gth)
er hand,
were blue
obbserved
e fibersin(~0.5
Saample
5 mm3 in and
leng4.gth)
These
were obbserved in Saam
observattions qualifyy in the abovvementioned
observatcriteria
tions qualify
(Norren,
y in 2007)
the abov
thherefore,
vementionedconnfirming
criteriathe
(Norpresence
ren, 2007) thherefore, connfi
 preliminnary study, further
f prelimin
reseaarch nary
on micr study, further
roplastics fcaan nowresea
bearch on micr
foocused onroplastics caan ation
thheir quantita now and
be foocused on thheir
characteerization usiing analyticcal tools suuch as the Fourier Trransform Innfrared SpecctroscopyTrransform Innfrare
characte
erization usiing analyticcal tools su
uch as the Fourier
Microsccope and Ram Microsc
man Spectrosscopy. cope
Micro and Ram
oplastic man
pres Spectros
sence needs scopy.
t be Micro
to oplastic
address sed bypressence
policy needs to
y makers t be addresssed b
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and stakkeholders. A suggested management p
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management p ESAMP,
program is 201
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5). in Fig. 4 (GE
ESAM
C. Ryan Argamino et al. / EnvironmentAsia 9(2) (2016) 48-54

Figure 4. Microplastic Managementt Figure 4. Microplastic Managementt

The first di imension off of The

themicroplastics
program m is first diimension
a Com mprehensive off the program m eration
is a Com mprehensive Waste Generati
therefore, confirming the presence particular shouldWaste Gen
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which M includes
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and the poopular 3R m
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Furtheer studies of pacckaging
either by mateerials
pollutant. In this case, microplastics ingested by significantly reducing the amount plastic used or
bivalves can be attributed to sources such as urban redesigning and formulating alternative materials which
inputs, fisheries, and aquaculture. Another aspect can be easily and safely degraded to non-threating
of CWGM is life cycle assessment which includes products (for both human and marine life) will decrease
on alterrnative packkaging materrials either by significaantly reduciing the amoount plastic used
value-chain
on alter models
rnative andpack the popular
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either byaltmarine litter output. Physical screening and waste
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h can amo ount
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roplastics. reducing Fiig. 5.Fig.shows 5. shows
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Figure 5. Comprehen
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Figure 5. Comprehen
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g maanagement plan
p
Another dim mension of the propossed management schem me is obtainning feedbaack f
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ks 51
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Fig. 6). It sttarts with proomoting awaareness on thhe impactsPeerception
oof these polllutant
and Mannagement)humans
(F
Fig. 6).
andIt the
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ng stakeholdders andoofmak
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psycholoogical aspecct of microplastic mannagement invplan for this soccial, behavio oral,
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9(2) (2016) 48-54

Figure 6. Risk Percepttion and Manaagement of Miicroplastics

Figure 6. Risk Percepttion and Manaagement of Miicroplastics
Another dimension on proposed
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to ensure Resources
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implemeented using the develo oped logies. Regu ulatory bodiesare a such being ass the Enviro onm
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impleme entedAnother
usingkey theaspect
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auditing bodentity
ies such to ensure
ass the that Enviro effluents
onmentaland
the risks involved is to convert complex terms marine bodies are checked in terms of microplastic
to a language that can be fully understood by the levels.
stakeholders. Developing a management plan for
this social, behavioral, and psychological aspect of 4. Conclusions
microplastic management involves surveys, psychological
studies, and acknowledging demographic differences. Microplastics were found present in Bacoor
These include regional, cultural, economic, educational, bay-cultured tahong (P. viridis). This preliminary
and other variances in behaviour and perceptions study confirms a microplastic pollution problem in
(GESAMP, 2015). Figure 5. Comprehen nsive wastethe g Bacoor ma
generation Bayanagement
and Manila plan
p Bay area and thus,
Figure 5. Comprehennsive waste generation g maanagement plan p
A comprehensive management framework is the need for further research on quantitating and
shown in Fig.Another
7. To mitigate the effectsAnother dim mension characterizing
of microplastic of the propos sed manage
these ment is ofschem me importance.
is obtainning feedbaack
dimmension of the propossed manage ment schem me pollutants
is obtainning utmost
feedbaack from
contamination to marine stakehol
and lders
human on
life, thei ir
policies perception n of
Management the risk ks
and posed by
mitigation microplastic
procedures c contaminat
to prevent ion (Risk Peerc
stakehollders on theiir perceptionn of the riskks posed by microplasticc contamination (Risk Peerception
should and Man nagement) (FFig. 6). It sttarts with pro omoting awa areness on thhe impacts o
of these pollluta
andbeMan
putnagement)
into place(F to strengthen
Fig. 6). It sttarts thewith
foundation
proomotingthe entry of
awaareness on microplastics
thhe impacts oof these (bothpoll primary
lutants toand
humans and the envvironment aliike. Educatinng stakehold ders and makking them unnderstand thee ha
of thehumans
measures andtothebe env
employed.
vironmentThe illustration
aliike. Educatinng also secondary)
stakehold ders and requiremakking them policy makers, the
unnderstand stakeholders,
e hazards
of microoplastic contaamination is an effectivee tool for maanagement (D Dippo, 2012)). Another keey
shows ofthe pillars
micro of mitigation
oplastic contaaminationactionisand an management
effectivee tool forgovernment maanagement agencies,
Dippo,non-government
(D 2012)). Another organizations,
keey aspect
of comm municating the t risks invvolved is to convert com mplex terms to a languaage that can be
whichofinclude
comm municating t risks invvolved
the
research, implementation, is to convertand
and regulation. com mplex
academic terms to a langua
institutions to age
workthat can be
in sync fullyup a
to put
understoood by the stakeholderss. Developinng a manageement plan for this soccial, behavio ora
understo ood by the
Research entities from the psycholostakeholders
government s. Developin ng a manage ement plan for this soc cial, behavio oral, and
ogical(Department
aspecct of micro strong
plastic andman effective
nagement management
invvolves surv framework
eys, psycho with
ological stud dies
psycholo
of Science ogical
and aspecct ofAcademic
Technology, microplastic mannagementresearch, invvolves surveys, psychoological stud dies, and
acknowlledgingInstitutions,
demographic diffferences. implementation,
Thhese include and r regulation
regional, as its
culltural, pillars
econom mic, educatio ona
acknowl
National ledging dem
Fisheries ographicand
Research diffferences.
Development Thhese include and regional,
r
policy as culltural,
its foundation. econom mic, educatio onal, and
other vaariances in beehaviour andd perceptions (GESAMP, 2015).
otherare
Institute,) vaariances
tasked toindevelop
beehaviour andd perceptions
infrastructure, methods, (GESAMP, 2015).

Figure 6. Risk Percepttion and Manaagement of Miicroplastics

Figure 6. Risk Percepttion and Manaagement of Miicroplastics
A compreheensive manaagement fram mework is shown
s in Fig. 7. To miitigate the effe
e
A compreheensive manaagement fram mework is shown
s in Fig. 7. To miitigate the effects
e of
microplaastic contam
mination to marine
m and huuman life, po
olicies shoulld be put into place to sttren
microplaastic contam
mination to marine
m and huuman52life, po
olicies shoulld be put into place to sttrengthen
the founndation of thhe measures to be emplooyed. The illu ustration also shows the pillars of miti
m
the founndation of thhe measures to be emplooyed. The illu ustration also shows the pillars of mitigation
m
action and
a managem ment which include reseearch, implementation, and a regulatioon. Research h e
action and
a managem ment which include reseearch, implementation, and a regulatioon. Research h entities
from thhe governmeent (Departm ment of Scieence and Technology, Academic IInstitutions, Na
l, and
s, and
al, and
Management Bureauu and Bureau u of Fisherie
Manage es and
ment Bureau Aquat
u and tic Resource
Bureau s will
u of Fisherie es act
andasAquat
an auditing entity
tic Resource to act as an audit
s will
C. Ryan
ensure thhat effluents and marine Argamino
bodies
ensure et al.
thhatare / EnvironmentAsia
chhecked
effluents andinmarine 9(2)
terrms of (2016)
micro
bodies are48-54
oplastic levells.
chhecked in terrms of microoplastic levells.

sed
cts ofor
o non-
ngthen
hysical
gation
s from
ntities
restrial
ational
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ntation
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Figure 7. Microplastic Management
t Framework
Figure 7. Microplastic Managementt Framework

4. Conclusions 4. Conclusions

Microplasticcs were founnd present

Acknowledgements in Bacoor
Microplastic cs werebay--cultured
Food foun tahhong
ndAgricultural
and present in (P. viriddis).
Bacoor This(FAO).
preeliminary
bay--cultured
Organization tahhong (P. viriddis). This
National
study coonfirms a miccroplasticstudy poollution probalem
coonfirms in the
miccroplastic
Bacoor
B Bay a
and Manila B
Bay area and
Aquaculture Sector Overview (NASO). Viale delle B
poollution prob lem in the Bacoor
B thus,
Bay a
andthe
Manila Bay area a
need for
r further rese
earch on
The authors would like to thankneed quaantitating and d
forr furtherGov-
the Provincial characteriz
reseearch zing
onTerme these
quaantitatingpo
ollutants
di Caracallaandd00153 is o
of
characteriz utmost imp
zing these
[homepage portance.
on thepoollutants
Internet]. is o
of utmost
Manage
ernment mentforand
of Cavite m
mitigation
assisting in the prrocedures
Manage ment to
procurement of prevent
and m the
themitigation e entry of microplastics
prrocedures
Rome, Italy: m2006;
to prevent
1: para s 28
(both prim
thee[cited
entry mary and19].
of microplastics
2016 m
Jan s (both
seconda
samples ry)Biology
and the requireLaboratory
p
policy maker rs, stakeholde
ofseconda
DLSU ers, policy
ry) providing
for require pgovernm ment
maker agencies
rs,
Available from:s,http://www.fao.org/fishery/countrysector/
stakeholde non-gover rnment
ers, governm mentorgan nizations,
agencies s, non-goverrnment or
and acad
the necessary demic
tools andinstitu
utions
reagents for to woork
and indemic
acad
conducting sync
the t put
to
institu
analysis. up a tostrong
s woorkand
naso_philippines/en.
utions e
effective
in sync t putman
to nagement
up a strong
s fra
amework
and e
effective mannagemen
with research, implementation, a resregulatio
and
with n as mentation,
earch, imple its GESAMP.
pillarrs and policy
a Sources,
and y as
fateits
regulatio found
nand
as dation.
effects
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