DS FishMetal

MARINDUQUE DEEP SEA SURVEY
JUNE 1998
FISH AND PRAWN
TISSUE CHEMISTRY
.
February 26, 1999
By: Joseph Mifsud

For: Placer Dome Technical Services (Philippines) Inc.
MARINDUQUE DEEP SEA SURVEY, JUNE 1998, FISH & PRAWN TISSUE CHEMISTRY 26/FEB/99
1.0INTRODUCTION
The Marinduque Deep Sea Survey was instigated by Placer Dome Technical Services Philippines
(PDTS) in June 1998, in response to public concerns that the dredging and submarine placement
of tailing trapped in the Boac River Channel will damage the deep sea fishing resource and
environment. A ten-day expedition collected baseline data on a previously unknown, deep sea
marine environment. This study looks at the distribution of metals in fish and prawn tissue
collected during this expedition.
To assess the deep sea fish resources five Z-traps were set in ten stations (from 400 to 750 m
deep) to capture fish and crustaceans off Marinduque Island, Philippines (WWC, 1999). Nine
stations were located in the potential submarine impact area, on the West Coast, and one
reference station in an area of no potential impact, on the South Coast, Figure 1.
Trawling was undertaken to assess the fishing resource in the potential impact area in the 60 to
80m depth range. Three trawls were attempted but only one was completed as planned, on two
other occasions the nets were destroyed during the trawl with reduced trawl time, Figure 1.
Fish and crustacean tissue sampling from four fish and one prawn species. I total; sixty-two tissue
samples consisting of flesh (caudal muscle) liver and whole prawns were collected. The species
used for tissue metal analysis were Synaphobranchus affinis (conger eel), Priacanthus tayenus
and P. macracanthus (big-eyes), Upeneus molluceusis (goatfish) and Heterocarpus gibbosus
(deep sea prawns).
The fish were dissected on board the research vessel TRV Sardinella, frozen and sent to Curtin
University in Western Australia, for chemical analysis. Fish caudal muscle and whole prawns were
used to simulate local eating customs and fish livers were used to assess the bioaccumulation
potential of heavy metals in these species. The results presented in this report provide new data
that is indicative of the metal body-burden in deep sea fish and prawns off Marinduque Island.
All fish flesh samples met the maximum residue level (MRL), a food guideline set by the
Australian National Health & Medical Research Council (NHMRC, 1989). Whole H. gibbosus
did not meet NHMRC guidelines, due to elevated median copper and cadmium concentrations.
However, the Marinduque prawns had lower copper and cadmium concentrations than Penaeus
esculentus (brown tiger prawn) caught in the Torres Strait and sold in Australia (GBRMPA,
1997).
Some individual liver samples from U. molluceusis did not meet the MRL due to elevated zinc
copper and selenium. However, the mean concentration of tissue metals did not exceed the MRL.
Fish liver is not a common source of human protein and is not normally marketed in Australia or
the Philippines.
The safe weekly fish and prawn consumption by an average 60 kg adult over a lifetime was
calculated using the permissible total weekly intake (PTWI) standards set by the World Health
Organisation (WHO 1972, 1973, 1986 and 1989). These calculations assume that the person is
not exposed to other sources of heavy metals. Our analysis shows that up to 26 kg of S. affinis,
43 kg U. molluceusis, 63 kg P. tayenus or 52 kg P. macracanthus flesh or 2 kg of whole H.
gibbosus can be consumed in one week, over a lifetime without any risk to health. Fish liver is
not normally consumed in local diets except for liver in very small dried fish. However, 1 kg of
U. molluceusis or 4 kg of P. tayenus or 8 kg of either P. macracanthus or S. affinis liver can be
/conversion/tmp/scratch/382652649.doc 1
consumed per week without any risk to health based on the WHO PTWI. These quantities of fish
tissue and whole prawns are not likely to be consumed by one individual every week, for a life
time.
2.0METHODS
2.1 Deep Sea Survey Site Selection
Fifteen stations were selected along the West Coast of Marinduque to represent; three depth
zones (78 to 230m, 340 to 443m and 537 to 748m), including a reference site on the south coast,
Figure 1. Fish where caught from the near-shore stations with a deep sea trawl net and from the
offshore stations with a Z-trap, (WWC, 1999).
Fish length and masses were recorded for each specimen. Scales were removed with a new
plastic knife on a stainless steel table. The cleaned fish was then transferred to a plastic foil were
the caudal muscle was exposed and dissected. The muscle tissue was washed using a squirt bottle
containing distilled water, excess water was then shaken off and the tissue placed in a plastic
ziplock bag. The abdomen was then cut open with a scalpel to expose the liver. This was
removed with clean surgical forceps, washed with distilled water, shaken dry and the liver placed
in a new plastic ziplock bag. Ten of the smaller goatfish livers were bulked into one sample.
Eel lengths and masses were recorded. The skin was removed with a sharp stainless steel knife. A
3 to 5 cm segment of caudal muscle was removed with a scalpel, washed with distilled water
shaken dry and the tissue placed in a labelled plastic ziplock bag. The liver was removed and
handled in the same way as fish liver.
Prawn lengths and masses were recorded before washing with distilled water, drip-dried and
placed in a labelled plastic ziplock bag.
All gross dissections were undertaken on board the research vessel and individually bagged
samples were placed in larger bags. All samples were stored in a freezer for one week before
shipping, to meet Australian Quarantine requirements. The frozen samples were transferred to ice
coolers with icepacks to keep the samples cold during shipment between Marinduque and Curtin
University in Perth, Australia. The Laboratory received the samples within 24 hours and in good
condition.
Sub-samples of tissue were dissected from the gross samples in laminar flow cabinets. These ultra
clean laboratory conditions ensured that contamination of samples was kept to an absolute
minimum. The tissue was homogenised and digested.
The digestion procedure:
1 Prior to the digestion stage the sample was reacted with redistilled nitric acid for 24h.
This facilitates the partial destruction of easily oxidisable material, to avoid the formation
of a “mousse” in the digestion vessel and the possible loss of sample due to frothing.
2 Next the sample was taken to dryness at 120oC to remove all easily oxidisable material. A
further 20mL of redistilled nitric acid was added and a watch glass placed over the mouth
of the digestion vessel.
3 The mixture was heated at 150oC under reflux for 48h with an additional 10mL redistilled
nitric acid added after the first 24h.
4 Next the sample was taken to dryness and a mixture of 20mL, 4:1 redistilled
nitic:perchloric acid was added. The mixture was refluxed at 170oC until clear or slightly
yellow.
Extreme care is exercised in the use of perchloric acid mixtures for the oxidation of
organic material, as there is considerable potential for the formation of explosive
perchlorate esters. The above procedures ensure that all oxidisable material is essentially
removed and the solution is capable of being heated to dryness without a risk of
explosion.
5 Finally the mixture was taken to dryness at 120oC taking care not to char the residue.
When dry, the final residue was taken up in 20mL of 5% redistilled nitric acid to await
further dilution prior to analysis. All samples were analysed within 48h of final
dissolution.
Sample preparation for mercury determinations was different from the total acid digest as this
element is volatile and its potential for loss under the previously detailed dissolution method is
high.
1 Approximately 100mg of sample were accurately weighed and mixed with 2g of mercury
free silica fume. The silica fume had previously been heated to 1,000oC for 24h to remove
all mercury.
2 The mixture was placed in a porcelain combustion boat in a tube furnace. The combustion
tube protruded from the furnace by 15cm before being restricted and angled at 90o. The
exit from the combustion tube was placed under the surface of 20mL of a mixture of 5%
sulphuric acid and 1% low-mercury potassium permanganate.
3 The sample was combusted in a stream of air (300ml min-1) at 700oC for five minutes after
which time total oxidation had been achieved. All exhaust gasses were bubbled through
the acid collecting solution, where the mercury was removed as mercuric sulfate.
4 5mL of this solution was then taken and reduced with hydroxylamine hydrochloride prior
to conventional cold vapour atomic absorption quantitation of the mercury concentrations.
The extract solutions from the digest were analysed using a Fisons PlasmaQuad3 Inductively-
Coupled-Plasma Mass-Spectrometer. All samples were diluted to give a final total dissolved
solids content of below 1000 ppm to avoid blocking the separation cones during long analytical
runs. Curtin University Laboratory conducts extensive quality control procedures and inter-
laboratory trials to ensure high quality analysis.
During the analysis of these samples, the laboratory effected drift corrections using repeat
standards every 12 samples; each analyte element was individually corrected for time/drift effects.
2.2 Data Management
Results from the chemical analysis were received in an Excel 97 format. Laboratory codes were
translated to site location, species and tissue sample and added to the field data, consisting of
length, weight and if the prawns were bearing eggs. The on-board weighing scales were
unrestrained and therefore affected by vibrations from the ship motors, all weight data is
presented to the nearest "stable" decimal unit.
Metal concentrations below the analytical method detection limits, were converted to the
detection limit for statistical purposes. The average (mean) was used as the measure of central
tendency unless the number of values below the detection limit were above 20% of the sample
size, in these cases the median was used. All statistical analysis was undertaken with Excel 97 or
Systat 7.0 software.
2.3 Quality Control
The following data quality objectives were adopted for this project.
1. The confidence interval (P=0.05) for through the run replicates must be less than 30% from
the mean, this allows for variability in results by samples that are close to the method
detection levels.
2. Relative percent difference for laboratory duplicate sample analysis was set at less than 35%
for results of more than 10 times the detection limit, or within one detection limit of each
other for results less than 10 times the detection limit, Gradient Corporation (1996)
3. Percent recovery of matrix spikes was set to within  25%.
4. Method blanks must be less than the detection limit for all analytes.
5. Analyses for standard reference material to be within statistically derived internal control
limits, set at  30%.
6. The detection limit for the laboratory methods used was 0.001 for all metals.
Certified dogfish liver tissue (DOLT) and reference muscle (DORM) were used to assess the
accuracy of laboratory methods, by measuring a known (certified) metal concentration. The
precision of laboratory analysis was assessed by incorporating a through the run single sample
replicate and a spot replicate every 10 samples.
3.0RESULTS
Table 1 presents a summary of the average metal concentrations in fish tissue and whole prawns
from the Deep Sea Survey. This table also shows the Maximum Residue Limit (MRL) set by the
Australian National Health & Medical Research Council for general food, fish, liver and
crustacean tissue, (NHMRC, 1989). The Philippines Bureau of Fisheries and Aquatic Resources
currently uses Tolerances or Guidelines from other countries such as Italy, Netherlands and the
U.K. The Australian standards selected for this study are generally more conservative and
comprehensive than the European guidelines used in the Philippines.
The complete tissue-metal concentrations are presented in Appendix A and summary statistics in
Appendix B.
TABLE 1: Average Metal Concentrations in Deep Sea Fish and Prawn Tissue off
Marinduque Island
DRY MASS, ppm Zn Cu As* Se Cd Pb Hg Fe Mn Cr Ni
Priacanthus tayenus, flesh (6) 8.608 0.654 1.424 0.040 0.001 0.018 0.000 43.519 11.319 0.153 0.109
Priacanthus macracanthus, flesh (1) 7.595 0.154 1.736 0.047 0.001 0.004 0.000 4.138 3.003 0.0365 0.024
Upeneus molluceusis, flesh (14) 7.419 0.735 2.088 0.097 0.001 0.025 0.001 58.805 3.754 0.127 0.103
Synaphobranchus affinis, flesh (1) 27.398 1.169 1.777 0.045 0.001 0.059 0.001 348.075 8.694 0.0631 0.042
Priacanthus tayenus, liver (6) 22.494 3.788 3.259 0.594 0.115 0.073 0.018 916.279 4.504 0.012 0.184
Priacanthus macracanthus, liver (1) 17.185 2.191 1.837 0.285 0.054 0.008 0.005 114.404 0.515 0.0003 0.007
Upeneus molluceusis, liver (4)+ 81.241 9.387 4.616 1.532 0.675 0.124 0.055 1453.208 4.683 0.001 0.958
Synaphobranchus affinis, liver (1) 16.399 2.025 7.987 0.042 0.018 0.007 0.000 173.584 0.727 0.0184 0.009
Heterocarpus gibbosus, whole (28) 9.827 21.162 12.099 0.056 0.221 0.067 0.001 352.650 37.875 0.152 0.250
NHMRC 1989, MRL - food [fish] 150 10 100* 1 [0.2] 1.5 [0.5] - - - -
NHMRC 1989, MRL - liver - - - 2 1.25 - - - - - -
NHMRC 1989, MRL - crustaceans - - - 1 0.2 - 0.5 - - - -
Penaeus esculentus, Torres Strait 32.41 22.94 14.46 0.98 2.3 0.06 0.02 31.34 0.77 0.13 0.28
Comparison to Torres Strait Study # -70% -8% -16% -94% -90% 12% -97% 1025% 4819% 17% -11%
(n) Numbers in brackets after the species name are the number of samples tested.
* MRL values are quoted for inorganic As, in this table we assume that 1% of total As is in an inorganic form.
+ A composite of ten small livers was treated as one sample, to be on the conservative side.
# P. Esculentus minus H. gibbosus metal concentration, divided by P. Esculentus.
The tissue metal concentration of H. gibbosus caught during this expedition were compared to
those of the Penaeus esculentus caught in the Torres Strait in a survey conducted by the Great
Barrier Reef Marine Park Authority (GBRMPA, 1997). Torres Strait P. esculentus are sold in
Australia. The last row on Table 1 shows the percentage difference in metal concentrations
between the H. gibbosus and the P. esculentus, a negative number indicates that the Marinduque
prawn has lower mean metal concentration than the Torres Strait prawn. Iron and manganese
were significantly higher in the Marinduque prawns but the potentially toxic metals selenium,
cadmium and mercury were lower in the Marinduque prawns. Copper, arsenic, lead, chromium
and nickel tissue concentrations were similar, that is within 20% of each other.
3.1 Quality Control
The analytical data met the project quality control specifications, established in section 2.3 as
follows:
1 The actual level of statistical variation of the throughout run single sample replicates
increases as the metal concentration approaches the detection limit for the analytical method.
For most metals within the normal range (ie. around 10 times the detection limit) Curtin
University was able to achieve around  10% variation, Table 2A.
2 Replicate sample analysis showed that the study's quality control standards were met for Ni,
Cu, Zn, Se and Cd. Two of the Hg and one of the Pb replicates did not meet the standards
because they were less than ten times the detection limit and more than one detection limit
apart, Table 2B. However, this should not significantly effect the interpretation of the results
for Pb and Hg.
3 Percent recovery of spike matrix was checked by internal laboratory controls and not
available for presentation. However, this statement is supported by the analysis of CRM’s
that show good recoveries of metal, see point 5 below.
4 Internal quality controls by Curtin University indicate that all method blanks were acceptable.
In practice our stated quality objective, of achieving less than the detection limit for all
analytes is difficult to achieve for tissue analysis because there is always some contamination
during digestion phase. However, all method blanks were very close to or below the
detection limit for all analytes (personal communication, Dr. J. Watling).
5 Analyses for standard reference material was within  30% or better, of the certified value,
Table 2C.
6 The detection limit for the laboratory methods used was 0.001 for all metals.
Table 2: Quality Control Test Results
DRY MASS Ni Cu Zn As Se Cd Hg Pb Fe Mn
Table 2A: Through The Run Single Sample Replicates
Average 0.359 7.899 18.783 0.115 0.064 0.049 0.008 0.020 19.045 104.906
Std dev 0.035 1.106 1.402 0.015 0.010 0.008 0.001 0.003 0.420 2.600
Confidence Interval 0.022 0.685 0.869 0.009 0.007 0.005 0.001 0.002 0.253 1.611
% from average 6% 9% 5% 8% 10% 10% 10% 10% 1% 2%
REPLICATE T1R1 0.383 7.263 16.556 0.114 0.049 0.046 0.005 0.020
T1R1 0.362 7.196 16.653 0.113 0.052 0.046 0.008 0.022
Test* OK OK OK OK OK OK Not 2 OK - -
REPLICATE T7R3 0.141 2.321 17.346 0.006 0.080 0.025 0.008 0.007
T7R3 0.184 2.539 17.158 0.007 0.094 0.028 0.011 0.008
Test* OK OK OK OK OK OK Not 2 Not 2 - -
REPLICATE 0F1R3 0.174 2.026 19.399 0.082 0.172 0.055 0.010 0.008
0F1R3 0.171 2.027 20.189 0.077 0.194 0.044 0.009 0.007
Test* OK OK OK OK OK OK OK OK - -
REPLICATE 20F0R3 0.148 4.634 16.937 0.052 0.063 0.058 0.007 0.020
20F0R3 0.145 4.759 17.524 0.065 0.069 0.045 0.008 0.020
Test* OK OK OK OK OK OK OK OK - -
DORM 1 1.242 5.186 21.146 17.076 1.895 0.076 0.801 0.396 64.5 1.24
1.235 5.282 21.283 17.453 1.742 0.071 0.837 0.366 64.7 1.30
1.198 5.219 21.528 17.240 1.733 0.070 0.778 0.334 65.2 1.30
CRM VALUES 1.200 5.220 21.300 17.700 1.620 0.086 0.798 0.400 63.6 1.32
Difference is < 20% OK OK OK OK OK OK OK OK OK OK
DOLT 1 0.266 21.256 89.255 11.186 8.266 4.624 0.236 1.856 718.0 8.51
0.243 20.943 90.043 10.696 7.738 4.862 0.235 1.788 709.2 8.49
0.252 20.499 90.243 10.425 7.853 4.590 0.213 1.564 710.3 8.84
DOLT 0.260 20.800 92.500 10.100 7.340 4.180 0.225 1.360 712.0 8.72
Difference is < 30% OK OK OK OK OK OK OK OK OK OK
* Item 2 in the quality objectives list Methods Section. OK means that the data quality is acceptable. Not 2
means the data quality is not acceptable, because the data is less than 10 times detection limit and the difference
between replicates is more than 1 detection limit.
4.0Discussion
The main concern for the submarine placement of dredge spoil with tailings, voiced by people of
Marinduque, is that the fish may become contaminated. This study establishes an indicative
baseline for metal concentrations in fish and prawns tissue found in deep waters near Marinduque.
The fish and prawns were caught at a considerable distance and depth from the Boac Dredge
Channel and are not likely to be contaminated by tailing.
Most tissue metal analysis met this study's quality control criteria, with the exception of two
replicates where mercury and lead concentrations did not meet condition 2 because they were
very close to the detection limit. However, these results have no significant effect on the
interpretation of the data discussed in this section.
Figures 2 to 8 compare the tissue concentration found in fish flesh, liver and whole prawns, for
seven selected metals with their respective MRL. All the fish flesh metal concentrations were
below the Australian MRL. Some individual U. molluceusis liver values, but not mean values,
exceeded the Se, Zn and Cu MRL.
The box plots show the median(middle line in the box) the first and third quartiles (bottom and
top box borders). The whiskers show a spread of 1.5 times the interquartile range and the values
falling outside this range are shown as stars or circles if more than 3 times the interquartile range.
The MRL values for each food category is show in red or if appropriate as a line across the page.
Figure 2: Total Tissue Arsenic

MRL food = 100 ppm
25
20
ppm (wet)
15
10
Species & Tissue
Figure 3: Total Tissue Cadmium

MRL liver = 1.25
1.2
1.0
0.8
ppm (wet)
0.6
0.4
MRL prawns = 0.2 ppm

0.2 MRL fish = 0.2 ppm
0.0
F L tF L L F L
W
Hg Pm Pm P Pt SaF Sa Um Um
Species & Tissue
Figure 4: Total Tissue Copper
40
30
ppm (wet)
20
10 MRL food = 10ppm
0
F L tF L L F L
W
Species & Tissue
Figure 5: Total Tissue Mercury

MRL fish and prawns = 0.5
0.150
0.125
0.100
ppm (wet)
0.075
0.050
0.025
0.0
W mF mL PtF PtL aF SaL mF mL
Hg P P S U U
Species & Tissue
Figure 6: Total Tissue Lead

MRL food = 1.5ppm
0.5
0.4
ppm (wet)
0.3
0.2
0.1
0.0
F L tF L L F L
W
Species & Tissue
Figure 7: Total Tissue Selenium
2 MRL liver = 2 ppm

ppm (wet)
1 MRL food and prawns

= 1 ppm
0
F L tF L L F L
W
Species & Tissue
Figure 8: Total Tissue Zinc
200
MRL food = 150ppm

150
ppm (wet)
100
50
0
F L tF L F L F L
W
Hg Pm Pm P Pt Sa Sa Um Um
Species & Tissue
The median copper and third quartile cadmium concentrations of whole H. gibbosus exceeded the
MRL for cadmium and copper. But both median concentrations were lower than those found in
prawns from the Torres Strait, Table 1. Copper is important for oxygen transfer in prawns and
tends to concentrate in the cephalothorax (GBRMPA 1995). MRLs are based on clean prawn
tails, which tend to have lower metal concentrations than whole prawns.
The MRLs essentially provide a guideline for food marketing in Australia, as the Torres Strait
study shows that exceedance of these MRLs is not necessarily a barrier to marketing of brown
tiger prawns in Australia. The percentage catch that would have technically met the Australian
MRL is presented in Table 3. The percentages in Table 3 are simply calculated by dividing the
number of individuals that exceed the MRL by the number of that total species analysed and
multiplied by 100.
Table 3 Percentage of the Marinduque Deepsea Catch that would have met NHMRC
1989 Maximum Residue Limit.
% of catch that was below MRL Zn Cu As* Se Cd Pb Hg

Priacanthus tayenus, flesh (6) 100 100 100 100 100 100 100
Priacanthus macracanthus, flesh (1) 100 100 100 100 100 100 100
Upeneus molluceusis, flesh (14) 100 100 100 100 100 100 100
Synaphobranchus affinis, flesh (1) 100 100 100 100 100 100 100
Priacanthus tayenus, liver (6) 100 100 100 100 100 100 100
Priacanthus macracanthus, liver (1) 100 100 100 100 100 100 100
Upeneus molluceusis, liver (4)+ 75 50 100 50 100 100 100
Synaphobranchus affinis, liver (1) 100 100 100 100 100 100 100
Heterocarpus gibbosus, whole (28) 100 14 100 100 54 100 100
NHMRC 85, MRL - food (fish) 150 10 10* 1 0.05 1.5 0.03
NHMRC 85, MRL - crustaceans 150 10 100* 1 0.2 1.5 0.5
(n) Numbers in brackets after the species name are the number of samples tested. * NHMRC value is for inorganic
As, this has been converted to total As by assuming that 1% of total As is in an inorganic form. + A composite of
ten small livers was treated as one sample.
Table 3 shows that all fish flesh metal concentrations meet the NHMRC criteria and are therefore
suitable for marketing in Australia. Technically only fourteen percent of the prawns met the
NHMRC Cu criteria for crustaceans and 54% meet the Cd criteria. However, the current metal
concentrations of whole prawns sold on the Australian market can be higher than those found in
Marinduque deep sea prawns, as demonstrated in Table 1 by the Torres Strait tissue metal study.
Cu, Zn and Se are essential dietary metals for humans. Cu toxicity is very rare in humans.
However, Cd may accumulate in human tissue.
Based on the 1989 NHMRC criteria, most fish livers, except U. molluceusis, were suitable for
marketing in Australia, if a market for fish liver existed, Table 3.
From a human toxicological perspective the most important standards are the Permissible Total
Weekly Intake (PTWI) proposed by the World Health Organisation (WHO). The PTWI levels
include all exposure sources but do not take metal interactions in a mixed metal matrix, into
consideration. However, they present the best guideline to assess how much an individual can
safely consume of a particular food in a week, based over a lifetime of exposure.
The "safe" total weekly intake of fish tissue and prawns were calculated from the WHO, PTWI
guidelines for a 60kg person. The calculations in Table 4 assume that there is no further exposure
from other sources of metal.
TABLE 4: Permissible Total Weekly Intake of Fish and Prawn Tissue from the Deep Sea
off Marinduque Island.
Kg of tissue safely consumed Zn Cu As* Cd Pb Hg

Priacanthus tayenus, flesh (6) 1626 321 63 1114 83 648
Priacanthus macracanthus, flesh (1) 1843 1361 52 402 418 620
Upeneus molluceusis, flesh (14) 1887 286 43 1502 60 331
Synaphobranchus affinis, flesh (1) 511 180 51 288 26 510
Priacanthus tayenus, liver (6) 622 55 28 4 21 16
Priacanthus macracanthus, liver (1) 815 96 49 8 179 55
Upeneus molluceusis liver (4)+ 172 22 19 1 12 5
Synaphobranchus affinis, liver (1) 815 96 49 8 179 55
Heterocarpus gibbosus, whole (28) 1425 10 7 2 22 580
WHO PTWI mg/60 kg/week 1,400 210 90 0.42 1.5 0.3
(n) Numbers in brackets after the species name are the number of samples tested. * WHO values are quoted for
inorganic As, this has been converted to total As by assuming that 1% of total As is in an inorganic form. + A
composite of ten small livers was treated as one sample, to be on the conservative side.
Fish are very seasonal food source and usually do not constitute the main component by weight of
a Filipino diet. The actual "safe" weight of tissue that can be consumed by an individual would be
lower, because in reality people are exposed to these metals from many other sources. In the
Philippines lead, cadmium and mercury exposures may be high due to vehicle emissions and high
tobacco consumption.
The calculations in Table 4 show that up to 63kg P. tayenus, or 52kg P. macracanthus, or 43kg
U. molluceusis, or 26kg of S. affinis flesh, or 2kg of whole H. gibbosus can be consumed in one
week without any risk to health. Up to 4kg of P. tayenus, or 8kg of either P. macracanthus or S.
affinis, or 1kg of U. molluceusis liver can be consumed based on the PTWI.
Cadmium was the most consumption quantity limiting metal in all the liver and prawn samples,
while arsenic was the most sensitive metal in finfish flesh metal and lead in eel flesh. It is unlikely
that more than 7 kg of fish flesh or prawns will be consumed in a week and even if the livers were
consumed with the fish no more than 700g of liver will be consumed in a week (that is 100g of
fish liver per day). Based on these assumptions we conclude that the deep sea fish and prawns
tested are suitable for human consumption.
4.1 Metal Distribution in Fish Tissue
Analysis of variance in the fish tissue metal concentration was undertaken for a number of factors
as summarised in Tables 5 and 6 below.
TABLE 5: ANOVA of Tissue Metal Concentrations in Deep Sea Fish and Prawns
Grouped by Species and Tissue.
Metal Species & Tissue

As HGW UML PTL UMF PTF
Cd UML HGW PTL PTF UMF
Cu HGW UML PTL UMF PTF
Hg UML PTL UMF HGW PTF
Pb NS
Se UML PTL UMF HGW PTF
Zn UML PTL HGW PTF UMF
Code: Factors are listed from highest concentration to lowest, reading left to right. Statistical significance for the
ANOVA analysis was set at P < 0.05; if P < 0.001 the factors are shown in blue. NS = not statistically significant.
Statistical significance for the factorial analysis using the Tuckey HSD test was set at P= 0.05. Factors joined by a
line indicate that they are not statistically different from each other. HG = H. gibbosus; PT = P. tayenus; UM = U.
molluceusis; W = whole prawn body; F = fish flesh (caudal muscle); L = fish liver.
This study found that prawns had significantly higher (P <0.0001) As and Cu concentrations than
fish tissue. High copper in prawns can be attributed to their use of haemocyanin (a protein
containing copper) for oxygen transfer in the blood compared to fish that use haemoglobin (a
protein containing iron). Most of the As in prawns may be attributable to sediment content in the
gut, which was not removed for prawn metal analysis.
U. molluceusis had higher Cd, Hg, Se and Zn than both fish flesh and prawns. P. tayenus liver
had significantly higher Se than fish flesh or whole prawns. These heavy metals usually
accumulate or are stored in liver tissue. Whilst there was no tissue isolation in prawns during this
study, GBRMPA (1997) found that heavy metals tend to accumulate in the hepatopancreas.
There was generally no significant difference in metal concentrations in fish flesh and liver
between trawl locations, Table 6, except for the following:
1. U. molluceusis flesh had significantly (P <0.05) higher As and Se and lower Pb

concentrations in trawl D3 than D1.
2. P. tayenus flesh had significantly (P <0.05) higher Cd in trawl D3 than D1.
The prawns caught in the deep sea Z-trap had similar tissue metal concentrations between sites,
with the following exceptions:
1. Significantly (P <0.05) higher Hg and lower As concentrations were found in T8 than the
other stations.
2. Significantly (P <0.05) higher Se tissue concentrations were found in T5 than in the other
stations.
Table 6 ANOVA of Tissue Metal Concentrations in Deep Sea Fish and Prawns
Grouped by Location.
Species P. tayenus U. molluceusis P. tayenus U. molluceusis H. gibbosus

Tissue Flesh Flesh Liver Liver Whole
Station T2, T5, T8, T11, T14
As NS D3 D1 NS NS T11 T5 T4 T2 T8
Cd D3 D1 NS NS NS NS
Cu NS NS NS NS T5 T11 T2 T14 T8
Hg NS NS NS NS T8 T14 T5 T11 T2
Pb NS D1 D3 NS NS NS
Se NS D3 D1 NS NS T5 T14 T2 T11 T8
Zn NS NS NS NS NS
Code: Factors are listed from highest concentration to lowest, reading left to right. Statistical significance for the
ANOVA analysis was set at P < 0.05; if P < 0.001 the factors are shown in blue. NS = not statistically significant.
Statistical significance for the factorial analysis using the Tuckey HSD test was set at P= 0.05. Factors joined by a
line indicate that they are not statistically different from each other. D1 = trawl TST12, D3 = trawl TST7; Tn =
trap followed by station number.
There were no statistically significant correlations between metal concentrations in fish tissue or
prawn with either length or weight.
5.0 Conclusion
An indicative baseline for tissue-metal concentration has been established, for five deep sea fish
and prawn species in the Marinduque deep sea habitat, the predicted final settlement area of the
dredge spoil. All fish flesh samples met the MRL guidelines for food in Australia.
Some individual fish liver samples from U. molluceusis and whole prawn samples did not meet the
Australian MRL guidelines. This study noted that fish liver is not normally consumed in the
Philippines except in very small dried fish, therefore the elevated Zn, Cu and Se found in a few
specimens will not present any health risks to humans. Whilst, the Marinduque prawns had lower
heavy metal concentrations than Torres Strait prawns, which are freely sold on the Australian
market, it would be advisable to exercise caution in consuming excessive quantities of whole deep
sea prawns from the Marinduque.
6.0References
GBRMPA, 1997, Heavy Metals in Commercial Prawn and Crayfish Species in the Torres Strait.
Great Barrier Reef Marine Park Authority, by Elizabeth Evans-Illidge, 1997.
Gradient Corporation, 1996, Snail Sampling and Analysis Plan. Marinduque Island,
Philippines. Gradient Corporation, Cambridge USA.
NHMRC, 1985, Metal Concentration Limits for Food. The 100th. Session National Health and
Medical Research Council, 1985.
NHMRC, 1989, MRL Standard. Standard for maximum residue limits of pesticides, agricultural
chemicals, feed additives, veterinary medicines and noxious substances in food.
Commonwealth of Australia.
NHMRC, 1990, The 1990 Australian market basket survey, National Food Authority, (Fay
Stenhouse), AGPS, Canberra.
WHO, 1972, Evaluation of certain food additives and the contaminants mercury, lead and
cadmium, WHO Technical Report Series No. 505 & FAO Nutrition Meetings Report
Series No. 51.
WHO, 1973, Trace Elements in Human Nutrition, WHO Technical Report Series No. 532.
WHO, 1986, Toxicological Evaluation of Certain Food Additives & Contaminants, WHO Food
Additives Series 21, Cambridge University Press.
WHO, 1989, Toxicological Evaluation of Certain Food Additives & Contaminants, Cambridge
University Press.
WWC, 1999, Deep Sea Survey, Marinduque Philippines, June 1998. A Baseline Survey of the
Marine Ecology in the Potential Impact Area of Dredge Spoil. Prepared by Woodward
Clyde (Philippines) for Placer Dome Technical Services (Philippines). {Draft}
Acknowledgments
Dr. J. Watling of Curtin University, Perth, for the metal analysis and technical support. Mr. R
Estudillo of Woodward-Clyde, Manila, for logistical support. The Crew of the Sardinella,
University of the Philippines Visayas, for their efforts in collecting samples under difficult
conditions. The PDTS Environmental Office Boac for sample handling, storage and shipment.
Appendix A
Tissue Metal Concentrations

Dry Mass (ppm)
Codes:
D1 Trawl TST 12, Figure 1.

D3 Trawl TST 6, Figure 1.
Tx Z-trap Station number x, Figure 1.
Hg: H. Gibbosus
Pm Priacanthus macracanthus,
Pt Priacanthus tayenus
Um Upeneus molluceusis
Sa Synaphobranchus affinis
F flesh
L liver
W whole
NB: Metal symbols are followed with their atomic weight.

Appendix B
Basic Statistical Analysis of Tissue Metal
Abbreviations:
#BDL, number of values below detection limit.

%BDL, percent of values below the detection limit.
Min, Minimum value.
Max, maximum value.
Median, a measure of central tendency.
Average, a measure of central tendency.

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MARINDUQUE DEEP SEA SURVEY

FISH AND PRAWN

February 26, 1999

By: Joseph Mifsud

2.1 Deep Sea Survey Site Selection

The digestion procedure:

2.2 Data Management

2.3 Quality Control

3. Percent recovery of matrix spikes was set to within  25%.

3.1 Quality Control

Table 2: Quality Control Test Results

Figure 2: Total Tissue Arsenic

Species & Tissue

Figure 3: Total Tissue Cadmium

MRL prawns = 0.2 ppm

Species & Tissue

Figure 4: Total Tissue Copper

10 MRL food = 10ppm

Species & Tissue

Figure 5: Total Tissue Mercury

Figure 6: Total Tissue Lead

Species & Tissue

Figure 7: Total Tissue Selenium

2 MRL liver = 2 ppm

1 MRL food and prawns

Species & Tissue

Figure 8: Total Tissue Zinc

MRL food = 150ppm

Species & Tissue

% of catch that was below MRL Zn Cu As* Se Cd Pb Hg

Kg of tissue safely consumed Zn Cu As* Cd Pb Hg

4.1 Metal Distribution in Fish Tissue

Metal Species & Tissue

1. U. molluceusis flesh had significantly (P <0.05) higher As and Se and lower Pb

2. P. tayenus flesh had significantly (P <0.05) higher Cd in trawl D3 than D1.

Species P. tayenus U. molluceusis P. tayenus U. molluceusis H. gibbosus

Tissue Metal Concentrations

D1 Trawl TST 12, Figure 1.

NB: Metal symbols are followed with their atomic weight.

Basic Statistical Analysis of Tissue Metal

#BDL, number of values below detection limit.

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