CR MANGROVE ROOT—~ E©&XxP. 2

2s 4
= 54
a
a
te]
a
Y ie]
Ww
u
= |
Zz
Cc
we |
-s
SEELEEEE EEL EERE
é 8 5
Fy

DAYS

<cR THALASS!(A ROOTS— EXP. 2

sod
= 4
fy ue 4
S 4
“A se4
Y 4
Wo 2p 4
v
z 4
ft ied
Ww
= 4

o 4

T r t T r 1
. é g 5
CRYS

Figure 30. Mean net uptake of Cr between experimental and control,
Thalassia roots and mangrove roots - Experiment II.

50


vTt

HOLOTHUR BODY EXPERIMENT NO=2

DAY NUMBER o

CADM TUM NO EXPERMENT MEAN= 1.97 DEVIATICN®

CHROMIUM NU EXPERMENT MEAN= 1.70 DEVIATIONS

COPPER NO EXPERKENT NE@l 4.45 DEVIATIUN®

NICKLE WO EXPERMENT WEAN 6.80 DEVIATION=

LEAD NO EXPERMENT MEANS 11.23 DEVIATIGN=

IC NU EXPERHENT MEANS 5.27 OEVIATIUAS
DAY NUMBER 1

CADMIUM T= 1.51 NOT SIGNIFICANT EME AN-CNEAN®

CHROM DUM EMEAN=CME AN

COPPER EMEAN®CKFE AN

NICKLE Te 1.64 NOT SIGNIFICANT EME AN-CME On=

LEAD Ts 5.41 SIGNIFICANT EME Aw-CME AN =

TINC Te 2.80 SIGNIFICANT EME AN-CHE Aiv =

. DAY NUMEER 5

CADMIUM T= 3.02 SIGNIFICANT EME AN-CME ANE

CHROMIUM T= 2.20 NOT SIGNIFICANT EME AN-CME AN =

COPPER EMEAN=CKEAN

NICKLE T= 5.79 SIGNIFICANT ENEA

LEAD T= 239° NCT SIGNIFICANT EMEA

TING Ts «70 NOT SIGNIFICANT EMEAW-CME ANS
DAY NUMBER 25

CADHIUM T= +98 NOT SIGNIFICANT EMEAN-CMEAN =

CHROMIUM T= 1.03 NOT SIGNIFICANT EMEAN-CHE AN®

COPPER Ts 1.00 NGT SIGNIFICANT EME An-CME ANS

NICKLE Te +95 NOT SIGNIFICANT EHEAn-CMEAN®

LEAD Ts 1.50 NOT SIGNIFICANT EME AN-CME AN]

ZINC T= 1.11 WOT SIGNIFICANT EMEAN-CMEAN®
DAY NUMBER 50

CADMIUM T= +37 NOT SIGNIFICANT EMEAN-CHE AN=

CHROMIUH T- 3.54 SIGNIFICANT EME AN~CHE AN®

COPPER tT +00 NOT SIGNIFICANT EMEAN-CHME AN=

NICKLE Ts +70 NGT SIGNIFICANT EMEAN-CMEAN®

LEAD Ts 1.81 NOT SIGNIFICANT EMEAWN-CME AN=

LINC Ts 1.36 NOT SIGNIFICANT EMEAN-CMEAN=
DAY NUMBER 65

CADMIUM T= 1.00 NOT SIGNIFICANT ENEAN-CNEAN=

CHROMIUM T= 71 NOT SIGNIFICANT EME AN-CME AN =

COPPER EMEAN=CMEAN

NICKLE Te 4.91 SIGNIFICANT EME AN-CME AN=

LEAD Te +29 NGT SIGNIFICANT EME AN-CMEAN=

ZINC T= 207 NOT SIUNIFICANT EMEAN-CMEAN=
DAY NUMBER 125 .

CADMIUM T= 26214.40 SIGNIFICANT EME AN-CME ANS

CHROMIUM T= 214.41 SIGNIFICANT EMEAN-CME AN®

COPRER EMEAN =CHE AN

NICKLE Ts 14.48 SIGNIFICANT EME AN-CME AN

LEAD Ts 1.98 NOT SIGNIFICANT EMEAN-CME AN=

ZINC Te 3.00 SIGNIFICANT EMEAN-CME SN&

15
+00
200
235
1.31
023

ol7

a7
2.83
247

tT
1.60

1.27
043
223

aT

3.63
1.75
71.40
17.07
1.30

05
62
+00
232
7.95
1.55

203
~.27

043
+10
203

220
7.27

5.20
1.83
9.43

DEG=

GEG=
DEGe
DEG.

DEG
DEG=

vEGTM
VEG
DEG

Deus
DEGe
OEus
DEG=
DEG=
DEG=

vEGs
CEGs
DEG=
DEGs
DEG=
DEG=

DEG=
OkGe

DEG
DEGs
DEG

DEG:
DEGe

DEG=
DEG
DEGs

wewwww

4

BRS PR PES

BE Vsuowy Heeeee

PDS

EMEANS

EMEAN®
EMEAN®
EMEAN®
EMEANS
EMEAN®=
EMEAN®

EMEAN=
EMEAN=
EMEAN®
EMEAN=
EMEANS
EMEAN=

EMEAN®
EMEAN®
EMEAN=
EMEAN®
EMEAN®
EMEAN®

1.63
1.70
4045
6017
11.43
4.60

1.20
3.13
e045
6.00
11.23
4.33

+63
5.10
6620
3.67

21.47

9-10

09S
4.65
G45
5.65
9.95
6.05

+73
5.37
4.45
6017
13.57
6.70

+80
3.97
4.45
11.27
7.20
19.33

EDEV=
eDE¥:
EvEV.

EDEV=

EDEVe

EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EOEVs
EDEVe
E0EV=
€DEV=

EDEVe
EOEV=
EDEV=
€DEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EOEV=
eDEV=

lS
1S
«lS
235
025
olT

00
1.24
1.24
20
247
215

21
60
3.03
2.06
16.31
82

a2

OF

CHEAN=
CHEAN=
CMEANE
CHEAN=
CHEAN=
CMEAN=

CHEANE
CMEAN=
CHEAN=
CHEAN=
CHEAW=
CHEAN=

CHEAN=
CHEAN=
CHEAN=
CHEAN=
CMEAN=
€MEAN=

CMEAN=
CHEAN=
CMEAN=
CMEAN=
CMEANS
CMEAN=

CHEANS
CMEAN=
CHEANS
CHEANS
CHEANS
CMEAN=

CMEAN=
CMEAN=
CHEAN=
CMHEAN®
CMEAN®

2-O0-CHEAN=

2

1.47 COEV=
1. 70CDEV=
4 -45CDEV=
5.70 CDEV=
8.60 CDEVe
4.33 CDEV=

1.47 COEV=
4.73 CDEV=
4.45CDEV=

7.27 COEV=
11.67 COEV=
4.10 COEV=

247 COEV=
6.73 CDEV=
4.45 COEV=
5.07 CDEV=
4.40 CDEV=
7.80 CDEV=

290 COEV=
3.83 CDEV=
4.45 CDEV=
5.97 CDEV=

17.90 CDEV=
6.50 CDEV=

«70 CDEV=
5.63 CDEV=
4.45CDEV=
6.60 CDEVe

13.47 CDEV=
6.67 COEV=

«60 CDEV=
1.70 COEV=
4-45CDEV=
6.07 COEV=
5.37 COEV=
9.90 CDEV=

12
00
200

226

37

223

1d
221
00
032
1.86


in the control tank. Obviously, metals were released from the
sludge, as shown by the results in Figs. 6 through 15 and, also,
by the significant net uptake of metals by Thalassia leaves and
roots, urchin and sea cucumber internal parts, and fouling or-
ganisms. This net uptake in organisms was hard to explain when
no net change was seen in the water of the experimental tank.

The problem in interpretation of the water results was due to

the "point sampling" of the water versus the "integrated"
sampling for the organisms, sediment, and sludge. The "point"
concentration of the water was very low at any one time (Appendix
C). These trace concentrations, coupled with the high blanks

and matrix problems in trace metal analyses of sea water by

flame atomic absorption spectrophotometry, produced results with
very high within-sample variance. This high variance effectively
obscured any differences between the experimental and control
tanks in Experiment II. Schuster and Pringle (1969) demonstrated
that the concentration of metal in the water, at any one time,
may not be the best indicator of the total contamination by metals.

The net uptake of metal by "fouling organisms" was clearly
shown in both Experiment I and II. This net uptake was replicated
in Experiment I and II for Cd, Pb, and Zn. The uptake of copper
by fouling organisms was only significant in Experiment I and
for Ni and Cr in Experiment II. The fact that Ni and Cr didn't
show a net uptake in Experiment I was probably due to a change
in sample technique for fouling organisms initiated in Experi-
ment II. In Bxperiment I, when a settling plate was sampled for
fouling organisms, clean plates were added for the next sampling
period, whereas, in Experiment II, a plate which was sampled
at day 125 had remained in the system from day 0 to day 125. The
fouling organisms sampled in Experiment II are, therefore, inte-
grated samples over the sampling time. The net uptake by the
fouling organisms of Cd, Pb, and Zn closely follows the loss of
these same metals from the sewage sludge for both Experiment I
and II. This same correlation, between fouling organisms and
sludge, holds true for Cd and Ni uptake in Experiment II.

The net uptake by Thalassia leaves and urchin internal
organs closely parallel each other for Cu, Cr, Pb, Ni and Zn
in both Experiment I and II. Significant cadmium uptake by
Thalassia leaves and urchin internal organs was seen gnly in
Experiment II. Schroeder (1975), using radioactive 57co, C0,
54mMn and ®5zn i.. sea water, showed that, in Thalassia testudinum,
the rhizomes had the highest uptake followed by leaves, then roots.
In both Experiments our results for Thalassia, show significant
net uptake for Cu, Cr, Ni and Pb in roots. The net uptake was
higher in Experiment.I for Pb and Cu and higher in Experiment II
for Ni and Cr. Cadmium and zinc had a net uptake in Thalassia
roots only in Experiment II. This uptake of metals by the
urchin and the close correlation between the urchin's uptake
and the uptake in Thalassia leaves was not surprising as the
urchins were voracious foragers on Thalassia leaves and epiphytes.

17


RESEARCH REPORTING SERIES

Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are

Environmental Health Effects Research

Environmental Protection Technology

Ecological Research

Environmenta! Monitoring

Socioeconomic Environmental Studies

Scientific and Technical Assessment Reports (STAR)
Interagency Energy-Environment Research and Development
“Special” Reports

9. Miscellaneous Reports

This report has been assigned to the ECOLOGICAL RESEARCH series. This series
describes research on the effects of pollution on humans, plant and animal spe-
cies, and materials. Problems are assessed for their long- and short-term influ-
ences. Investigations include formation, transport, and pathway studies to deter-
mine the fate of pollutants and their effects. This work provides the technical basis
for setting standards to minimize undesirable changes in living organisms in the
aquatic, terrestrial, and atmospheric environments

ONONSBwWH =

This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.


OTT

URCHIN SHELL
DAY NUMBER °
CADMIUM T= 287 NOT SIGNIFICANT
CHROMIUM T= +78 NOT SIGNIFICANT
COPPER EAEAN =CHE AN
NICKLE Te 1.03 NGT SIGNIFICANT
LEAD Te 014 NGT SIGNIFICANT
ZIKC Te 1629 NOT SIGNIFICANT
DAY NUMBER 1
CADMIUM T= 2.06 NOT SIGNIFICANT
CHRUMIUM T= +86 NOT SIGNIFICANT
COPPER Ts 200 WOT SIGNIFICANT
NICKLE T= 1.74 NOT SIGNIFICANT
LEAD Ts 2.36 NOT SIGNIFICANT
ZINC Ts +51 NOT SIGNIFICANT
DAY NUMBER 5
CADMIUM Ts 1.92 NOT SIGNIFICANT
CHROMIUM T= 3.74 SIGNIFICANT
* COPPER Ts 200 NOT SIGNIFICANT
NICKLE T= 1.68 NOT SIGNIFICANT
LEAD Ts 3.53 SIGNIFICANT
ZINC T= 1.09 NOT SIGNIFICANT
DAY NUMBER 25
CADMIUM T= 033 NOT SIGNIFICANT
CHROMIUM T= -69 NOT SIGNIFICANT
COPPER EMEAN =CHE AN
NICKLE Te «55 NOT SIGNIFICANT
LEAC Ts 2.47 WOT SIGNIFICANT
ZINC T= 5.01 SIGNIFICANT
DAY NUMBER 5u
CADMIUM T= 2.83 NOT SIGNIFICANT
CHROMILM T= 1.00 NOT SIGNIFICANT
COPPER EMEAN =CNE AN
NICKLE T= of6 NOT SIGNIFICANT
LEAD Ts +23 NOT SIGNIFICANT
ZINC ENEAN®CME AN
DAY NUMBER 125
CADMIUM T= 1.26 NOT SIGNIFICANT
CHROMIUM T= 18022.40 SIGNIFICANT
COPPER EMEAN=CMEAN
NICKLE Te «90 NOT SIGNIFICANT
LEAD Ts 6-84 SIGNIFICANT
ZINC Te 13.50 SIGNIFICANT -

EXPERIMENT NO=1

EME AN-CME Avs
EME AN-CMEAN®

EME AN-CME AN =
EME AN~CME Aw
EME AN-CME AN=

EME AN-CMEAN®
EMEAN-CME AN
EME AN-CME AN =
EME Aiv-CME AN =
EME AW~CME AN®
EME AW-CME An =

ENE AN-CME Aly
EME AN-CME ANS
EME AN-CME ANS
EME ANW-CHE ANS
EMEAN-CME AN®
EME AN-CMEAN=

EME AN-CMEAN®
EME An-CidE ANS

EMEAN-CHE AN®
EME AN-CME AN®
EME AN-CKE AN=

EME AN=CME AN®
EME AN-CHE ANS

EME An-CME AN=
EMEAN-CME AN=

EME AW-CHE AN®
EME Au~CME AN =

EWE AN-CME AN=
EME ANCHE AN=
EME AN*CME AN®

1.47
7620

10.95
0
55

3.09

3.54

+20
1.10

“95
6.75
2.70

DEG=
OEG=

DEGs
VEG=
DEGe

DEGs
DEGs
VEGs
DEue
veGe
OEus

vEGs
OEGe
VEGe
DEGe
OEG=
vEGs

DEG=
DEGS

VEGe
DEG
DEG=

DEGs
DEGs

DEG=
DEG=

OEG=
DEG=

DEG=
OEG=
DEGs

RN ORR RRR ORR PEERS ES VUE AN ND

new

RN

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN®

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®=
EMEAN=

EMEAN®=
EMEAN=
EMEAN®
EMEAN=
EMEANS
EMEAN®

EMEAN=
EMEANS
EMEAN=
EMEAN®
EMEANE
EMEAN®

1.82
4.65
4045
31.65
43.70
11.45

3.85
3.00
445
10.10
44666
15.54

215
5.60
4645
4.60

56.35
11.55

3.90
3.50
44S
31.90
12.10
10.50

4025
6655
4645

15.40

4.75
6.50
4645
27.90
56.35
16.20

EDEVe
EDEV=
EDEVe
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV®
EDEV=
eDEV=
EDEV=

EDEVe
EDEVe
EDEV=
EDEVe
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

eDEVs
EDEV=
EDEVe
EDEVe
EDEV=
EDEV=

2.37
235
3s
15.06
12.45
o2l

2.37
224

200
5-28
1 7
8.11

“200

hs

26

CHEAN®
CMEAN=
CHEAN]
CMEAN®
CHEAN=
CHEAN=

CMEANE
CMEAN®
CMEAN=
CMEAN=
CHEAN=
CHEAN®

CHEAN=
CHEAN=
CHEAN=
CHEAN=
CMEAN=
CHEAN=

ChEAN=
CHEAN=
CKEAN=
CHEANS
CMEAN®
CMEAN=

CMEAN=
CNEAN=
CMEAN=
CMEAN=
CHEANS
CMEAN=

CMEAN=
CMEAN®
CMEAN=
CMEAN®
CMEAN®
CMEAN®

20

3.30 COEV=
4.85 CDEV=
445C0EVE

22.66 CDEV=
42.50 COEV=
12.00 CDEV=

015 COEV=
3.80 COEV=
4045 COEV=
3.20 COEVe

63.05 COEV=
12.45 CDEV=

3.69 COEV=
2.60 COEV=
4.45 CDEV=
18.95 CDEVe

42.80 CDEV=
9.80 COEV=

4.00 CDEV=
2.65 COEV=
4.45CDEV=
33.35 CDEV=
10.30 CDEV=
13.20 CDEV=

4.85 CDEV=
6-50 CDEV=
4.45CDEV=
2 5 CDEV=
56.35 CDEV=
15.40CDEV=

4.55 CDEV=
5.40 CDEV=
SCDEV=

26.95 COEV=
51.60 CDEV=
13.50 COEV=

1.48
200


CO HOLOTHURIAN GUT Exp 2

N
1

MEAN <E-C) ,PPM
5

=4
& -_
& Fy
CAYS
CR HOLOTHURIAN GUT- EXP 2
sa
= 4B
fom
‘ 3a
A
o oe
ly
‘ois
Zz
tal a
=
-1
a
e T
& z 5
cAYS

Mean net uptake of Cd and Cr between experimental and

Figure 39.
control, sea cucumber guts - Experiment ITI.

59


AANAMAAANANDADAAAAANAAADANANAADAAANAANDANANAANAANANANANANANABANANAA

AAD WD DYDD WWW DG WWW Ot et et et tet tet net tet tt tet tt ed tet aa tn Det feat Dat tent at end at Head Det ted teed fd tt tad Pd od tt
COP RP RP RP ODO OME RH OOOO RP RP RE HOBO O OR HH OOOrF HOO RP HOOrHMOOOr HE HHO

ee NN ol ol el el al

SORTED DATA 1

0 24 -999.9 31.6 -999.9 -999.9 81.0

0 2.7 -999.9 31.4 -999.9 -999.9 83.0

0 -999.9 -999.9 17.5 -999.9 -999.9 108.2
tt) 2.0 0 186.4 607 4.5 167.6

0 9 1.7 14.4 6.9 5.0 89.4

0 1.0 1.1 17.4 7.9 5.9 107.4

1 -999.9 -999.9 10.6 -999.9 -999.9 164.7

1 999.9 -999.9 10.0 -999.9 -999.9 164.3

1 9 =-999.9 7.0 7.5 6.5 191.2

1 -999.9 -999.9 21.9 -999.9 -999.9 270.1

i 4.2 -999.9 26.2 -999.9 -999.9 306.9

5 999.9 -999.9 11.2 -999.9 10.2 216.3

5 -999.9 -999.9 9.9 -999.9 14.1 241.9

5 3.7 -999.9 20.6 -999.9 -999.9 211.4

5 -999.9 -999.9 19.6 -999.9 -999.9 216.6
25 2.0 -999.9 9.8 -999.9 -999.9 187.2
25 2.0 -999.9 12.3 Gol -99909 184.6
25 -999.9 -999.9 27.4 -999.9 -999.9 85.2
25 2.0 8 15.2 10.4 626 101.8
50 2.4 3.7 9.7 9.4 5.6 187.1
50 3.5 5.8 8.5 5.0 10.5 194.0
50 1.2 -999.9 9.5 7.6 10.0 168.3
50 5.0 3.5 10.6 14.1 12.7 353.6
50 3.6 3.0 725 11.3 10.7 268.3
50 1.5 1.9 5.8 926 6.1 187.5
125 1.6 1.4 5.5 5.6 5.8 139.6
125 1.6 2.1 4.8 2.0 4.4 109.6
125 1.3 -999.9 4.7 248 5.3 9947
125 1.4 -999.9 6.0 6.8 5.9 119.0
125 1.4 -999.9 6.4 6.6 5.8 114.6
125 2.0 -999.9 1.6 2.0 602 69.8
125 2.0 2.8 52 5.0 602 119.5
125 1.6 2.5 5.1 7.9 4.0 120.5
125 1.5 3.1 5.3 726 4.7 117.1
0 4.0 -999.9 1.2 -999.9 20.8 1.7

0 4.0 -999.9 1.4 -999.9 20.0 2.6
i?) 3.3 6.7 6el 32.9 49.8 3.0
ie) 3.1 0 0 33.8 20 3.6

9 4.0 -999.9 1.4 -999.9 10.5 3.3

Q 3.0 -999.9 164 -999.9 13.4 1.5

0 3.4 1.4 5.6 33.7 49.6 4.6

0 344 3.1 54 34.0 54.5 52

1 -999.9 -999.9 1.4 -999.9 13.5 1.6

1 4.6 5.2 0 36.9 13.3 5.5

1 3.2 6.4 6.0 34.0 57.0 3.3

i 3.1 6.8 bel 34.0 5309 3.6

1 -999.9 -999.9 1.30 -999.9 11.4 05

1 -999.9 -999.9 1.2 -999.9 15.9 3

1 3.1 6.9 62 33.7 51.0 4.2

1 3.1 6.9 602 34.0 51.9 0

5 999.9 -999.9 2.4 -999.9 20.0 1.3

5 999.9 -999.-9 1.6 -999.9 11.5 9

70


taken: pore water for analysis of soluble trace metals and hydro-
carbons, overlying water for soluble and particulate trace metals
and hydrocarbons, and dissolved reactive phosphate (DRP). The
DRP was sampled at weekly intervals. The trace metals and hydro-
carbons in water and pore water were sampled at the same inter-
vals as the organisms. Pore water samples were not taken in
Experiment II.

Pore Water

Samples were taken using one-inch PVC pipes sealed at one
end with 0.8 mm holes drilled in the lower 38 cm of the pipe.
The pipes were placed 46 cm from the sides at 1.2, 3.5, 5.8, and
8.1 m from the intake end of the tanks. Two liter water samples
were withdrawn using 6.4 mm Tygon “~ tubing and 25 lbs of vacuum

pump.
Sediment

Sediment was collected using a 2.5 cm diameter x 25 cm
PVC core. The sediment core was homogenized and ground after
drying (105°C) and a 1 g sample taken for trace metals. After
day 25 in Experiment I and throughout Experiment II, a 5 g sample
was taken for trace metals.

Dissolved and particulate trace metals

All water samples were placed in 2.5 & glass bottles
(which formerly contained conc. HNO, or HC&) which were
equilibrated in reagent grade sea water for 12 hrs prior to
sampling (4 £ samples for Experiment II). The sample bottles
were drained, rinsed with the sample, filled, then filtered
within 4 hrs. The filtering was accomplished using an all
plastic filter system containing a .4 ym pore size, 142 mm di-
ameter, Gelman ® filters. The filters were previously washed
in 6 N HC and rinsed with copious amounts of R.0O./D.I. water.
The fraction retained on the filter was referred to as the
"particulate" portion. The filtrate was the "soluble" fraction.

Physical and Chemical Analyses
General

All reagents used are reagent ACS grade. All glassware
was cleaned with detergent, rinsed with water from a Millipore
"Milli-Q" system. All water referred to in this paper is R.0./
D.I. water from the Millipore "Milli-Q" system. This refers
to the technique of reverse osmosis followed by ion-exchange.
The final cleaning for glassware used to digest samples for
trace metal analysis involved refluxing the covered beakers
with concentrated HNO, followed by rinsing with R.0./D.I. water.


6TT

MANGROVE

ADM I1UK
CHROMIUM
COPPER
NICKLE
LEAD
TING

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHRGRIUM
COPPER
NICKLE
LEAD
ZING

ROOT EXPERI
DAY NUMBER 0
NO EXPERMENT MEANS +80
NO EXPERMENT HEAN= 24.37
NO EXPERMENT HEAN= 10.37
NO EXPERHENT MEAN= 32.57
NO EXPERMENT MEAN= 4.23
NO EXPERMENT KEAN= 22.97
DAY NUMBER = 5
Tz 10.84 SIGNIFICANT
Ts 1.53 NOT SIGNIFICANT
Ts 1.41 NGT SIGNIFICANT
Ts 18.59 SIGWIFICANT
Ts 2612 WOT SIGNIFICANT
= 14.76 SIGNIFICANT
DAY NUMBER 50
EME AN =CME&N
Te 10.13 SIGNIFICANT
Ts 19.63 SIGNIFICANT
Ts 11.23) SIGNIFICANT
Te +93 NOT SIGNIFICANT
Ts 5.76 SIGNIFICANT
DAY NUMBER 125
Ts +87 NOT SIGNIFICANT
Te 80.30 SIGWIFICANT
EMEAN=CMEAN
Te 2.35 NOT SIGNIFICANT
Ts 4.49 SIGNIFICANT
Ts 1.72 NOT SIGNIFICANT

MENT NOS2

DEVIATION=
DEVIAT Une
DEVIATION=
DEVIATIUN=
DEVIATIONS
DEVIATIUN=

EME AN-CME AN®
EME AN-CHE AN
EME AN-CME AN =
EME AN-CME AN=
EME AN-CME AN >
EME AN-CHE AN =

EMEAN-CMc AN®
EME AN-CME AN =
EHEAN-CHE AN=
EME AN-CME AN=
EME AN~CHME AN®

EME AN-CME AN =
EME AN-CME AN=

EME AN-CHE ANE
EME AN-CMEAN=
EME AN-CME AN =

+20
3.70
221
1.17
42
2.48

7647
1.12
2.76
9.73
2.99
85

9.13
5

6.00
72.68
3.50

7210
23.33

1.33
5.77
3.60

NUM=
KUK®
UKs
RUMS
NUM=
hUK=

DEG=
DEG=
OEG=
DEG=
DEG=
DEG=

DEG=
DEG=
DEG=
DEG
DEG=

DEG=
DEG=

DEGs
OEGs
DEG

wEe eww

weuuwe

Repes

BP

Pas

EMEAN®
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS

EMEAN=
EMEANS
EMEAN=
EMEAN?
EMEAN=
EMEAN®

EMEAN®
EMEAN®
EMEAN=
EMEAN®
EMEANS
EMEAN=

EDEV=

7 EDE¥=

EDEVe
EDEV=
EDEV=
EDEV=

EREV:
EDEVe
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

CHEAN®
CHEAN=
CMEAN=
CHEAN=
CMEAN=
CHEAN>

CMEANS
CMEAN=
CMEAN®
CMHEAN=
CMEAN=
CMEAN>

CHEANS®
CHEAN=
CHEAN=
CMEAN=
CMEAN=

~CHEANS

29

1.00
16.45
7.07
16.90
2.07
17.05

COEV=
COEV=
COEVe
COEV=
CDEV=
CDEV=

33CDEV
7.73
4.45
4.47
5.23
12.43

+80
7.20

+45CDE 00
12.77 CDEV= +96
+53 COEV= 2.22

18.73 COEV=


SEWAGE

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM LUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

DAY NUMBER

NO EXPERMENT
ND EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
DAY NUMBER

NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
DAY NUMBER

NG EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
DAY NUMBER

NO EXPERMENT
NO EXPERMENT
NO EXPERMENT

0

MEAN=
MEAN=
MEA
MEAN=
MEAN=
MEANS
1

MEAN=
MEAN=
MEANS
MEAN=
MEAN=
MEAN=
5

MEAN=
MEAN=
MEAN=
MEAN=
MEAN=
MEAN=

25

MEAN=
WEAN=
MEAN=

NG EXPERMENT
NG EXPERMENT
NO EXPERMENT
DAY NUMBER
NO EXPERMENT
NG EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NG EXPERMENT
DAY NUMBER
NG EXPERMENT
NG EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT MEAN=
NO EXPERMENT MEAN=
DAY NUMBER 125

NO EXPERMENT MEAN=
NOG EXPERMENT MEAN=
NO EXPERMENT MEAN=
NO EXPERMENT MEAN=
NO EXPERMENT MEANS
NO EXPERMENT MEAN=

65
MEAN=
MEAN=
MEAN=
MEAN=

EXPERIMENT NO=2

165.97
3666.87
2445.33

375.60

21.40
2507.60

58.33
1567.57
1374.40

279.40

21.03

1193.60

58.97
1979.43
1016.17
324.40
16.17
1261.57

51.57
1845.97
1306.80

245.43

34.03

1205.63

66.50
1979.07
1449.63

229.60

57.17

1444.57

31.13
1253.47
884.53
237.60
77.57
857.17

17.97
658.73
502.50
139.57

19.90
431-40

125

DEVIATION=
DEVIAT ION=
DEVIAT 10H=
DEVIATION=
DEVIATION=
DEVIATION=

OEVIATION=
OEVIATION=
DEVIATION=
OEVIAT ION=
DEVIAT 10N=
DEVIATION=

DEVIATION=
DEVIATION=
DEVIATIU
DEVIATIO
DEVIATIG
DEVIATION=

DEVIATION=
DEVIATION=
OEVIATION=
DEVIATIO
BEVIATION=
DEVIAT ION=

DEVIATION=
DEVIATION=
DEVIATION=
DEVIATION=
DEVIATION=
DEVIAT I0N=

DEVIAT ION=
DEVIATIONS
DEVIATION=
DEVIATION=
DEVIATION=
DEVIATION=

DEVIATION=
DEVIAT ION=
DEVIATION=
DEVIATIGAN=
DEVIATION=
DEVIATION=

85.45
336.46
266.43

12.67

3.29
122.13

6.07
272.14
183.97

36.28

4.87

154.57

5013
198.34
36.58
15.70
4024
278.32

5.55
307.45
304.00
18.41
14.56
238.87

8.32
141.86
37.69
12.66
5.46
14.36

8.56
32.51
9.31
17.57
16.23
70.63

1.04
8.11
57.23
931
00
22.85

Vw wwu

weEwWUWW

VWOWWWYW WHWEWH LBWHLWL WWLLWYW WHwuUDYD


Huggett, R.J., M.E. Bender, and H.D. Slone. 1973. Utilizing
metal concentration relationships in the eastern oyster
(Crassostrea virginica) to detect heavy metal pollution.
Water Research 1: 451-460.

Ikuta, K. 1968. Studies on the accumulation of heavy metals
in aquatic organisms. II. On accumulation of copper and
zine in oysters. Bull. Jap. Soc. Sci. Fish 34: 112-116.

Jackim, E. 1973, Influence of lead and other metals on fish 6-
Aminoleuilinate dehydrase activity. G. Fish. Res. Board
Can. 30: 560-562.

Jacobs, S.A. 1973. Some measurements of heavy metal content of
sewage effluent and sludge. In: The use of flowing biological
systems in aquaculture, sewage treatment, pollution assay,
and food chain studies. Woods Hole Oceanographic Inst.

Tech. Rept. WHOI-73-2. January.

Jones, J.A. 1968. Primary productivity by the tropical marine
turtlegrass, Thalassia testudinum Kénig, and its epiphytes.
Ph.D. thesis, University of Miami, Coral Gables, Florida
January 1968.

Kelly, J.A., C.M. Fuss Jr., J.R. Hall. 1971. The transplanting
and survival in Boca Ciega Bay, Florida. Fishery Bulletin
69(2): 273-280.

Kerfoot, W.B. and S.A. Jacobs. 1973a. Premeditated cadmium
contamination of a combined waste water treatment. Aqua-
culture system In: The use of flowing biological systems
in aquaculture, sewage treatment, pollution assay, and
food chain studies. Woods Hole Oceanographic Inst. Tech.
Rept. WHOI-73-2. January.

Kerfoot, W.B. and S.A. Jacobs. 1973b. The accumulation of metal
by organisms cultured in a combined tertiary treatment-
aquaculture system. In: The use of flowing biological systems
in aquaculture, sewage treatment, pollution assay, and food
chain studies. Woods Hole Oceanographic Inst. Tech. Rept.
WHOI-73-2. January.

Kerfoot, W.B. 1973. Cadmium accrual in a flowing marine microcosm.
In: The use of flowing biological systems in aquaculture,
sewage treatment, pollution assay, and food chain studies.
Woods Hole Oceanographic Inst. Tech. Rept. WHOI-~73-2. January.

Menzel, D.W. 1977. Summary of experimental results: Controlled
ecosystem pollution experiment. Bull. Mar. Sci. 27€1)
142-145.

Montgomery, J.R., S.E. Kolehmainen, M.D. Banus, B.J. Bendien,
J.L. Donaldson.and J.A, Ramirez. 1976. In: National Bureau
of Standards special publication #422 Accuracy in Trace Analysis:
Sampling, sample handling, and analyses. 7th IMR Symposium
Gaithersburg, MD. p. 163-172.
65


Addition of sewage
!

t
Collection of sediment/ Collection of water
sludge/organisms (0,1, samples for trace
5,25,50,85**, 125 days) metal and hydro-
+ carbon analysis
sample taken*
Frozen in plastic bags for analysis Filtered
of hydrocarbons
Defrost Frozen

sample for* hydro- R
Dissection carbon analysis Amberlite XAD-2
resin columns and

. R
Sample pooled by tissue Extracted with CCL, Chelex-100 resin

css Extract stored in column
Subsample* freezer
mercury
analysis Elution, dilution
wet weight |
Analysis, atomic
Dry 105°C absorption
Dry weight
Grind by
mortar/pestle
Digest/dilution
!
Analysis,

atomic absorption

Samples were collected at 0,1,5,25,50,85** and 125 days after addition of
sewage sludge.

*Samples for mercury, and hydrocarbons in both water, sediment, sludge and
organisms were stored frozen for future analyses.

**An additional sampling was made for Experiment II, Day 0 refers to the sam-
ple taken before addition of sewage sludge.

Flow chart of trace metal and hydrocarbon sampling
from collection processing into final sample form.
23

Figure 3.


v6

EXPERIMENT RO=1

HOLOTHUR BODY
DAY NUMBER )
CADMIUM T= 8.50 SIGNIFICANT
CHROMIUM EHEAN®CMEAN
COPPER EMEAN®CME AN
NICKLE EMEAN=CREAN
LEAD Ts 3.94 NOT SIGNIFICANT
Tine Ts 1.57 NOT SIGNIFICANT
DAY NUMBER 1
CADMIUM T= 205 NOT SIGNIFICANT
CHROMIUN EMEAK=CMEAN
COPPER EMEAN=CKMEAN
NICKLE Ts +89 NOT SIGNIFICANT
LEAD Te 233 NOT SIGNIFICANT
ZINC Ts 1.03 NGT SIGNIFICANT
DAY NUMBER 5
CADN LUM = 1-15 NOT SIGNIFICANT
CHROMIUM EMEAN =CME Aiv
CCPPER Te 200 NOT SIGNIFICANT
NICKLE Ts 3 NGT SIGNIFICANT
LEAD Ts 13.82 SIGNIFICANT
ZINC Te 2.99 SIGNIFICANT
DAY NUMBER 25
CADMIUM T= 3.05 NOT SIGNIFICANT
CHROMIUM EME AN =CMEAN
COPPER EMEANSCKE AN
NICKLE T 5.330 SIGNIFICANT
LEAD T= +38 NOT SIGNIFICANT
THC Ts 3.14 NOT SIGNIFICANT
DAY NUMBER 50
CADMIUM T= 1.46 NOT SIGNIFICANT
CHROMIUM EMEAN=CKEAN
COPPER Te 1.63 NOT SIGNIFICANT
NICKLE T= «62 NUT SIGNIFICANT
Leap Ts 1-06 NOT SIGNIFICANT
ZINC Ts +25 NOT SIGNIFICANT
DAY NUMBER 125
CADMIUA Te 2.65 NOT SIGNIFICANT
CHROMIUM EMEAN®CMEAN
COPPER Ts 200 NOT SIGNIFICANT
SICKLE Ts 200 NOT SIGNIFICANT
LEAD Ts «13 NOT SIGNIFICANT
ZINC Te 1.12 NOT SIGNIFICANT

EMEAN-CME AN =

EME Alv-CME AN =
EME AN-CME AN =

EME AN-CME AN =

EME AN-CME AN =
EME AN-CME AN=
EME AN-CME AN =

EME Aw-CME AN=

EME AK-CME AN =
EME Aw-CME AN =
EME AN-CME AN =
EME ANCHE AN=

EME Aw-CMt AN=

EME AN-CME AN=
EME AN-CME AN=
EME AN-CHE AN =

EME AW-CME AN =

EME AN-CME AN
EME AN-CME AN=
EME AN-CNE AN =
EME AN-CME AN =

EME AN~CHE ANS

EMEAN-CME AN
EME AN-CHE ANS
EME AN-CME AN =
EME AN=CME AN®

95

DEG=

nn

x

72.57

2
2
2

Pos

wwoe wo

EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
ENEAN=
EMEANS
ENEAN=
EMEAN >
EMEAN=

EMEAN=
EMEANS=
EMEAN=
EMEAN=
EMEAN=
EMEAN®

EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEANS

EMEANE
EMEAN®
EMEAN®
EMEANS
EMEAN=
EMEAN®

EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=

1.45
1.70
4645
11.95
7.00
18.05

+82
1.70
4045
10.40
7.05
10.65

1.90
1.70
4645
10.43
11.97
12.62

1645
1.70
445
411.95
7.60
9.95

2.40
1.70
4.45
9.23
7.77
15.37

+90
1.70
4045

EDEV=
EDEV=
eDEV=
EDEV=

EDEV=

“EDEV=

EDEV=
EDEV=
EDEV=

EDEV=

EDEV=

EDEY=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EvEV=

EDEV=

EDEV=

EDEV=

EOEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=

700-£DE¥=

11.25
7.00

EDEV=
EDEV=

207 CHEAN=
007) CMEAN=
07 CHEAN®
-07  CMEAN=
1.27 CMEAN=
2-47 CMEANS

295 CMEAN=
95
295
2.12
6.01
1.77

+64 CHEANS
264 CMEANS
200 CMEAN=
45 CMEAN=
+49 CMEAN=
1.96 CMEAN=

+21 CHEANS
«21 CMEAN=
e21) CHEAN=
1.34 CHEAN=
6.65 CMEAN=
+07 CHEANS

224 CMEAN=
024--CHEAN=
+00 CHEAN=
1.33 CMEAN=
S96 CHEAN=
9.75 CHEAN=

700 CHEAN=
00) CHEAN=
200 CMEANS
00. CHEAN=
1.06 CHEAN=
+71 CHEANS

3

2.40 CDEV=
1.70CDEV=
4.G5CDE Ve
11.95CDEV=
10.55 CDEV=
22.70 CDEV=

087 CDEV=
1.70CDEV=
445CDEV=
12.00 CDEV=
5.50 CDEV=
9.00 CDEV=

1.35 COEVe
1.70CDEV=
4.45 COEV=
1 O CDEV=
6.45 CDEV=
8.10 CDEV=

+90 COEV=
1. 70COEV=
4eGSCOEVE
6.55 CDOEV=
5.80 COEVs
6.65 CDEVe

5.90 COEV=
1.70CDEV=
7Tel2 CDEVe
10.00 CDEV=
3.05 COEV=
"13.55 CDEV=

2.10 CDEV=
1.70CDEV=
4.45 COEV=
7.65 CDEV=
411.47 CDEV=
9.57 COEV=



oe

Figure 10.

PB FOULING ORGANISMS— ©&xP. 2

sea 4
ue 4

3B 4

MEAN <E-C> » PPM

284
a214

DAYS

Mean net uptake of Pb between experimental and control, fouling organisms - Experiment II.


SS

Figure 35.

ZN MANGROVE ROOT- ©&xP 1

es,
= 78
. a.
Qa
ar
Y vel
ld
Y
<
4
c
y mo 4
-s TT 7 T Tr T T T 7
5 : : T
Ss i g
CRYS

Mean net uptake of Zn between experimental and control, Mangrove roots

~- Experiment I.


CD FOULING ORGANISMS— ©&XP. '

Pee |
= 4
a
a2]
‘
~ 4
y |
tJ
~ 4
Zz -2
c
Wa 4
=
Soe
T T T T T T r
= é z Fy
DRYS
<D SEWAGE—- EXP. i
2s 5
12@ 4
=
a7 4
a
r4
oc se 4
=
2 4
a T T T T T T 1
3
& g Fi
CAYS

Figure 6. Mean net uptake of Cd between experimental and
control, fouling organisms and sewage - Experiment I.

26


CR HOLOTHURIAN GUT—- EXP. 1

MEAN <E-C) / PPM

ig

CAYS

NI HOLOTHURIRAN GUT- EXP. 1

2 |
= 4
a
Qu 4
.

.
Yo |
ls
wo |
zd
Cc
ud 4
=
-= 1
— ——

CAYS

Figure 38. Mean net uptake of Cr and Ni between experimental and
control, sea cucumber guts - Experiment I.

58


S6

HOLOTHUR

CADMIUM
CHROMIUM
EGPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM IU
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
WECKLE
LEAD
ZINC

INTERNAL
DAY NUMBER 0
Ts 1.04 NOT SIGNIFICANT
Te 4.49 SIGNIFICANT
Ts +47 NOT SIGNIFICANT
Ts 1.75 NOT SIGNIFICANT
Te 2.96 SIGNIFICANT
Ts 2.82 SIGNIFICANT
DAY NUMBER 1
1 1.69 NOT SIGNIFICANT
Ts 205 NUT SIGNIFICANT
Ts 291 NOT SIGNIFICANT
Ts 3.33 SIGNIFICANT
T= 22.17 SIGNIFICANT
Ts 4.17) SIGNIFICANT
OAY NUMBER = 5.
Ts +22 NOT SIGNIFICANT
Ts 5.91 SIGNIFICANT
T= 86425.60 SIGNIFICANT
Te 15.39 SIGNIFICANT
Ts 4.35 SIGNIFICANT
Tz 27.66 SIGNIFICANT
DAY NUMBER 25
Ts 1.72 NOT SIGNIFICANT
Ts 15.47 SIGNIFICANT
Ts 16.31 SIGNIFICANT
Ts +12 NGT SIGNIFICANT
1: 18.8 SIGNIFICANT
Ts 11424 + SIGNIFICANT
DAY NUMBER 59
Ts 1.96 NOT SIGNIFICANT
1. 13.2 SIGNIFICANT
Ts 743 SIGNIFICANT
Ts 1.26 NOT SIGNIFICANT
T= 22.33. SIGNIFICANT
Te 9.03 SIGNIFICANT
DAY NUMBER 125
Te +71 NOT SIGNIFICANT
Tx 67637) SIGNIFICANT
Ts 4.48 SIGNIFICANT
Ts 11.30 SIGNIFICANT
Ts 14,89 SIGNIFICANT

Ts 9.62

SIGNIFICANT

EXPERIMENT NO=1

EME AN-CME AN=
EME AN-CME AN =
EME AN-CHE AN®
EAE AN-CME AN =
EMEAN-CME ON
EME AN-CAE AN

EME AN-CHE AN =
EME AN-CME AN =
EME ALI-CME AN =
EME AN-CME AN=
EE AN-
EME Ali

EME AN-CHE AN =
EME AN-CME EN =
EME AN-CME AN
EME AN-!
EME AN-CME @l
EME AN-CME AN=

EME AN-CME AN =
EME AN-CME AN
EME AN-CME AN
EME AN-CME AN=
EME AN-CME AN=
EME AN-CME AN=

EME Aiv-CME AN =
EME AU-CME ANS
EME Aw-CHE AN =
EME AN-CME AN
EME AN-CME ANS
EME AN-CME AN =

EME AN-CME AN=
EME AN~CHE AN =
EME AN-CME AN=
EME AN-CME AN
EME AN-CME AN=
EME AN-CME AN =

= 655
-11,02
71.66
“5.74
~b 41
10.40

1.17
-.10
2.26
6.82
13.35
26.05

025
7.75
10.55
8.50
18.56
28.00

1.45
14.38
19.90

+43
49,83
29.43

1.08
20.2

27.4.

1,28
65.72
50.10

--40
11.85
11.95
=4.55
45.6

18.47

PRUNE VEL EY AAKARKN UROUEW

pupess

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS

EMEANS
ENEAWS
EMEAN=
EMEANS
EMEANS
EME ANS

EWEAL
EMER
EMEANS
EMEANS
EMEAN=
EMEANS

EMEANS
EMEAKS
EMEANS
EMEANS
EMEAN=
EMEANS

EMEAW=
EMEAN

EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=

1.15
7.50
7.27
18.50
Geld
32.10

2.10
9.80
10.03
14.53
16.75
60.70

1.07
13.90
15.00
24.85
21.55
56.15

3.57
26.00
29.80
21.90
63.20
65.30

3.20
29.90
37.35
22.45
79.30
79.75

2.35
29.65
26.80
23.85
68.15
62.60

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
eDEV=

EDEV=
EOEV=
EVEV=
EDEVe
Ebev=
EDEV=

EDEV=
EOEV=
cDEVs
EDEV=
EDEV=
EDEV=

EbEV=
EDEV=
EDEV=
cOEV=
EDEV=
EDEV=

EDEV=
eDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEVs
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

22k
71
4.00
1.98
2.47
8.20

52
2.50
212
1642

7.88

1.3)
1.41
00
07
6.01
78

CHEAN=
CMEAN=
CHEANS
CMEANS
CMEAN®
CHEANE

CMEAN=
CMEAN=
CMEAN=
CMEAN=
CMEAN=
CMEAN=

CHEAN=
CMEAN=
CMEAN=
CHEANS
CHEAN=
CMEAN=

CHEAN=
CMEAN®
CNEAN=
CAEAN=
CHEAN®
CMEAN®

CHEAN®
CHEAN=
CHEAN®

JCHEAN®

CHEAN®
CHEANS

CHEANS
CMEAN®
CHEAN®
CHEANS
CMEAN®
CMEAN=

4

1.70
18.52
8.94
24.24
10.56
42.50

92
9.90
7.77

21.35
3.40
34.65

+82
6.15
4645
16.35
3.05
28.15

2.11
11.62
9.90
21.47
13.37
35.87

2.12

9.7

9.95
21.17
13.87
29.65

2.75
17.80
14.65
28.40
22.6

4G.12

COEV=
COEVe
COEV=
COEV=
COEV=
CDEV=

CDEV=
CDEV=
CDEV=
CDEV=
CDEV=
CDEV=

COEV=
CDEV=
COEVE
COEV=
CDEV=
CDEV=

COEV=
CDEV=
CDEVe
CDEV=
COEV=
CDEV=

COEV®
COEV®
COEV=
COEVe
COEV=
COEV=

COEV=
COEV=
COEVs

COEV=



aa}

MANGROVE STEM EXPERIMENT NG=2 27
DAY NUMBER 0

CADMNIUK «ONO EXPERMENT MEAN] 240 DEVIATIUN= +10 3
CHROMIUM NO EXPERMENT MEAN= 0 DEVIATION= 60 3
COPPER NO EXPERMENT HEAN= +45 DEVIATIONS +00 3
NICKLE NO EXPERMENT KEAN] 4.07 DEVIATIONS +32 3
LEAD NO EXPERMENT MEAN= 3.72 DEVIATION 3.04 3
ratte NO EXPERMENT MEAN= 4.67 DEVIATICNE 15 3
DAY NUMBER 5
CADMIUM T# 1.80 WOT SIGNIFICANT EMEAN-ChE AN= ~e15 DEG= 3 EMEAN= 255 EDEV= +07 CHEAN= 70 COEV= +10
CHROMIUM T= 1,80 NOT SIGNIFICANT EMEAN-CMt ON= 1.88 VEG= 3 EMEAN= 7.65 EDEV= 1.48 CKEAN= 9.73 COEV= 93
COPPER «Te 200 NOT SIGNIFICANT EMEAN-CME AN = 200 LEG= 3 EMEAN= 4645, +00 CHEANS 4.65 COEVE 200
NICKLE Ts 5.28 SIGKIFICANT EME Aw-CME FI 6.85 DEG= 3 ENEAN= 12.95 2.33 CMEAN= 6.10 COEV= +56
LEAD Te 13.83) SIGNIFICANT EME Ais-CME Ai 6.70 DEG= 3 ENEAN= 6.95 +92 CMEAN= «25 CDEVE 00
ZINC Te 2.55 WOT SIGNIFICANT EME AN-CME At 6.20 VEG= 3 EMEAN= 8.30 EDEV= +85 CMEANE 14.50 CDEV= 3.21
DAY NUMBER 50
CADMIUM EMEAN=CHEAN EMEAN= +15 EDEV= -85  CMEAN= +1SCOEV= 200
CHROMIUM EMEAN=CHE AN EMEANS 1.70 ivEV= +85) CHEANS 1.70CDEV= +00
COPPER EMEAN=CKE AN EMEAN= 4.45 EDEV= +85 CMEAN= 4eG5COEV= 00
NICKLE T= +73 NOT SIGNIFICANT EHEAN-CME AN = -.27 DEG= 4 EMEAN= 2.60 EDEV= ~53 CHEANS 2.87 CDEVe 35
LEAD EME AW SCME AN EMEAK= +25 EvEVs +53 CMEAN= +25CDEV= 00
ZINC Ts 1.86 NOT SIGNIFICANT EME AN-CMEAN= -290 VEG= 4 EMEAN= 4.30 EDEV= +70 CMEAN= 5.20 COEV= 346
DAY NUMBER 125
CAOHIUH Te 248 NOT SIGNIFICANT EMEAN-CME AN +05 DEG= 4 EMEAN= +40 +00 CMEAN= +35 COEVe +18
CHROMIUM T= 210 WOT SIUNITFICAKT EMEAGW-CME An= +10 DEG= 4 EMEANS 3.10 1.23 CMEAN= 3.00 CDEVe 1.14
COPPER Ts 1.00 NOT SIGNIFICANT EWE AN-CME AN= 1.55 DEG= 4 EMEAN= 6.00 2.68 CHEAN= 4.45 COEVE 200
WICKLE T= 223 NOT SIGKIFICANT EME AN-CME AN= -.07 DEG= 4 EMEAN= 4el3 +32 CHEANS 4.20 CDEV® 240
LEAD Ts 1.00 NOT SIGNIFICANT EMEAN-CME AN= 2.98 VEG= 4 EMEAN= 3423 EDEV= 5.17 CHEAN= +25 COEY= 10
Tine Ts 1.63 NOT SIGNIFICANT EMEAN-CMEAN= -1.43 LEG= 4 EMEAN= 7.77 EDEV= +65 CMEAN= 920 CDEV=



CU THALASSIA LERVES- ©&xP 1

MEAN ¢E-C)> PPM

DAYS

CU URCHIN GUT- EXP. 1

MEAN ¢E-C) /PPM

aq
@h4
28
aziy

CAYS

Figure 16. Mean net uptake of Cu between experimental and control,
Thalassia leaves and urchin guts - Experiment I.

36


Woh tat tah be Bet ttt tet ak ek fed he Dat bat et et tt bt ba he bet tet et tt bet bt tt Bet tat et tet Bet Bet bt bet dt
HOOP MOO HE HOO HH ODDO QOH HOODH EEE OO DOO OPP HE MOOR HE HODOOF

cececceccececccecccceeccecceccececececceccecececececcececece
YO UUNUHY WWD DUG AYD UY DADDY DW et et ne tt nt et tet ce nt tt att tt toe teat ttt mt et tt tt

SORTED DATA 9
5

4.5 10.8 9.5 23.6 16.7 40.3

25 3.0 27.9 22.0 39.9 37.9 91.5
25 3.4 29.9 25.9 45.9 37.1 92.1
25 6.3 23.5 20.2 36.6 39.6 85.9
25 4.8 18.6 19.9 35.8 39.8 9401
25 3.4 14.0 14.0 21.9 16.6 53.9
25 3.3 13.5 19.3 20 18.2 50.2
25 5.7 9.7 10.5 23.2 14.0 40.6
25 5-9 9.7 11.1 23.0 16.0 44.9
50 202 36.6 24.9 5348 29.5 92.7
50 264 3604 27.0 57.9 32.7 96.8
50 3.5 27.9 20.0 50.7 20.3 20
50 344 2965 19.0 51.0 20.4 20
50 4.6 34.7 17.3 44.7 19.9 7764
50 4.6 34.7 17.4 44.6 19.8 77.3
125 2.6 22.9 2769 18.9 55.9 166.4
125 2.3 18.9 22.9 17.0 46.6 207.0
125 4.6 19.3 19.0 26.1 50.0 172.9
125 4.6 22.6 19.3 25.4 49.8 187.8
125 4.5 18.6 18.7 24.8 49.6 179.1
125 4.3 19.6 19.4 24.8 49.7 171.2
125 3.4 7.6 Tel 18.9 21.0 89.9
125 3.7 6.5 8.0 19.0 19.6 114.6
125 6.1 8.2 9.0 16.5 20.4 123.2
125 6.2 6.7 8.8 16.3 20.4 118.2
0 -999.9 4.4 1.0 1.0 34.9 11.3

0 3.5 4.9 4.3 22.3 52.5 11.6

0 3.5 4.9 4.1 22.9 43.0 12.4
0) 3.1 4.8 4.1 22.3 42.0 11.6

1 -999.9 4.4 1.0 8.5 56.8 11.5

1 2.8 3.8 1.0 3.9 5561 94

1 544 2.9 4.5 7.0 37.9 8.4

1 5.8 3.3 4.2 16.4 35.6 26.2

1 5.1 3.4 3.9 14.7 37.9 2202

1 -999.9 3.8 1.0 3.9 64.1 12.4

1 -999.9 3.8 1.0 2.5 62.0 12.5

5 -999.9 5.3 a] 5.2 56.2 10.4

5 -999.9 5.9 1.0 4.0 56.5 12.7

5 3.9 3.4 4.6 8 44.8 12.4

5 -999.9 3.6 9 Te8 491 10.1

5 5.5 2.6 4.4 15.6 38.1 8.3

5 52 2.8 4.2 17.6 39.2 8.4
25 3.9 3.5 6.1 30.9 12.8 10.0
25 3.9 3.5 Sel 32.9 11.4 11.0
25 3.7 2.8 4.4 30.9 10.5 13.0
25 4.3 3.6 5.1 35.86 10.1 13.4
50 4.4 6.5 6.2 28.9 57.8 14.9
50 4.1 6.6 6.2 27.8 57.8 15.9
ET) 5-0 6.5 6.2 29.8 60.7 15.9
50 4.7 6.5 602 27.9 56.0 14.9
4.8 6.5 6.6 27.8 57.7 16.4

4.7 6.5 5.3 28.0 59.0 16.0

oo
we
uN
ww

78


ACKNOWLEDGEMENTS

The authors would like to express their appreciation to
the following organizations: the Puerto Rico Department of
Agriculture for wet laboratory facilities and Harbor Branch
Foundation and the Smithsonian Institution for financial support
and use of facilities for completion of this report. The
technical support of the following people was invaluable for
the completion of this report: Mr. Pedro Acosta, Mr. Jon Cole,
Mr. José Echevarria, Mr. Thomas Prinslow, Mr. José Ramirez,
Ms. Arlene Ramfrez, and Dr. Kenneth Watters. Special thanks
go to Mrs. Barbara Herman, Mrs. Jackie McKay, and Mrs. Rose
Neville for typing the manuscript, and Mr. Tom Smoyer for
photographic aid. Our sincere thanks to Dr. Frank Lowman for
the initial conception of this project.


FOREWORD

The Environmental Research Laboratory of the U.S.
Environmental Protection Agency is located on the shore of
Narragansett Bay, Rhode Island. In order to assure the
protection of marine resources, the laboratory is charged
with providing a scientifically sound basis for Agency
decisions on the environmental safety of various uses of
marine systems. To a great extent, this requires research
on the tolerance of marine organisms and their life stages as
well as of ecosystems to many forms of pollution stress. In
addition, a knowledge of pollutant transport and fate is
needed.

The waters of coastal marine environments, as well as
the inshore sediments, often become the ultimate sinks for
pollutants released into them from industrial sources.
Plants and animals in those areas are subjected to varying
concentrations of the contaminants which can be taken up,
transported, and made available to upper trophic levels
including man. Studies as the one reported herein help to
develop and test predictive models on the bioavailability,
uptake, and transport of toxic substances in marine tropical
systems.

This small scale experiment serves as an initial step
to assess the impact of pollutants in larger aquatic
environments.

Eric D. Schneider
Director
Environmental Research Lab. Narragansett , RI

iii


NI FOULING ORGANISMS— EXP. 2

MERN <E-C) » PPM

a
Bh)
Ls)

#21

DAYS

Ni SEWRGeE- Exe. 2

sae 5
aso |
3me 4
zse |
zee |

isa 4

MERN PPM

Mean net uptake of Ni between experimental and control,

Figure 13.
fouling organisms and sewage - Experiment Ii.

33


BIBLIOGRAPHY

Camp, D.K., S.P. Cobb, J.F. VanBreedveld. 1973. Overgrazing of

seagrasses by a regular urchin, Lytechinus variegatus.
Bioscience 23(1): 37-38.

Carmody , D.J., J.B. Pearce and W.E. Yasso. 1973. Trace metals
in sediments of New York Bight. Pollut. Bull. 4(9): 132-134.

Davey, E.W. and A.E. Soper. 1975. Apparatus for the sampling
and concentration of trace metals from sea water. Limnol.
Oceanog. 20(6): 1019-1023.

Eisler, R. 1971. Cadmium poisoning in Fundulus heteroclitus
(Pisces: Cyprinodontidae) and other marine organisms.
J. Fish. Res. Bd. Canada 28: 1225-1234.

Eisler, R. 1973. Annotated bibliography on biological effects
of metals in aquatic environments (no. 1-567). EPA report
no. EPA-R3-73-007, Corvallis, Oregon.

Eisler, R., G.E. Zaroogian and R.J. Hennekey. 1972. Cadmium

uptake by marine organisms. J. Fish. Res. Bd. Canada 29:
1367-1369.

Ferrell, R.E., T.E. Carville, and J.D. Martinez. 1973. Trace
metals in oyster shells. Environmental Letters 4(4): 311-316.

Frazier, J.M. 1976. The dynamics of metals in the American
oyster, Crassostrea virginica II. Environmental effects.
Chesapeake Sci. 17(3): 188-197.

Gardner, G.R. and G. LaRoche. 1973. Copper induced lesions in
estuarine teleosts. J. Fish. Res. Bd. Canada 30: 363-368.

Goldberg, E.D. 1965. Review of trace element concentrations in
marine organisms Vols. I & II. Puerto Rico Nuclear Center.

Golley, F., H.T. Odum, and R.F. Wilson. 1962. The structure
and metabolism of a Puerto Rican red mangrove forest in May.
Ecology 43(1): 9-19.

Hannan, P.J. and C. Patouillet. 1972. Effect of pollutants on
growth of algae. Report on NRL progress.

64


NOMA ADND ARR DEDNON ANMOMTNADONDANMNE ER NODODADETAMAMOMAMNIN
Sete ee te tm eee ewe cose eee ewer ewer ee eee eee seseseseserere
PEE NORE ONARAN DAFA ST TON FAN TAANNANNMONMNNOMANMH TOOTS TeOaRdaAra
TOODDOOMN THO SN SIA n - -
QOUVAVANGOMAAMN DAM TINARARAKRAHONHROONAKRHNNADORNGOCOOHRAROOS
Cre ae a ae ee a re ee rer area ae arrears
DASHA NAANAHOGAR  AAATRTAARAMAAAMFAANAAAROTFARHAT ARORA TE
Atettet tet AAR ARARHRATARHR BF DADHRAAA ao
BA STHSSTARAR FO RAKSAH ao LES
' trrrrarag ove treba wo rit
ANPONRDRNONORWROHRM ONY VOETOCAOCCOOVCOVMAMNT + ONMOOOTAOCOOMENOS
cee eee eee eae err ee ceed err et ieee are rer re ke
ASMANMARMORONODAGH AAA mao NAANNN ONAN OA TAM
i er)

CAM STM AVTORNAVOONNARRAGTOODNHODAN NE OTONADONMTHRODRINANENOOMS

cee re ee er eer ara ee eee eee eee eee era

AI EMNAN ORE SASK EMA MNNAANNAANANAM MO MONG OS ONES TE SANE

+
DRAMA NS VY GON DDN AAA HAH NAN MD NROMMN ANH D ANNA AAA KAA AKAD
ce ee pier rae ee a a a Cae ee
Facaam nogamnoen AHRAMAKA HF AAROANM SRPFARAAABRARAAAAHRAG
PADD a ANRTARAHPA TARAS AA BAGTARRAARADAAD
BARS HHAKPFAAKS AHRAK FR BEDASASAAKHAAGN

err rrrrrrertareta rt oprrrrrr rrr ernst
<«

=

ELIA Moonsrsarennenqnnyns sr ornnenannsv easseedsvanyon
cee. Ce ee i ee a ae er cee ce ee
APOAMBHANN AHA 7 a aa atte aoa

a AR O

LD An w

me trot

«

MMNNDNNHODOOHNHNNN O©OOOGSOHHMNDDIOONNESCOSONNODOONNHNHNNNHOOS
NNN ARK AANA NN WANNA NS DANA wa

ttt aa atest

ADOMADDDDOO OH MANHOOD OHANOOH HOOF NAMA O OMA HONOO NH AMOOHMHOORHOOR
EE NN EE NN Nt LL tt et tt et et tt et tt tt et ot
SEAABTTTST ATS TT TFMMADAGMADDADAMDMDAAMAMDVOOVOVOHVOYVYVYUYVOUSSAddH
BETTE TTETTCFLLC CLC LCF LS TTFSFLTLTLLTLEAATTTSETFLLKRTFTTLAEKLATTETZATEUISTE

73


United States Environm ental EPA-600/3-78-035
Environmental Protection Research Laboratory March 1978
Agency Narragansett Rl 02882

Research and Development _

SEPA iological Availability
of Pollutants
to Marine Organisms

Ecological Research Series

>



66

MANGROVE
DAY NUMBER
CAOMIUK Te 4062
CHROMIUM
COPPER Te +00
NICKLE Ts 00
LEAD
2Ine Ts 12.89
DAY NUMBER
CACHIUM T= 1.41
CHROMIUH
COPPER
NICKLE 1s 6.06
LEAD Ts 2.63
ZINC Te 10.11
. DAY NUMBER
CADHIUK Te 2.83
CHROMIUM
COPPER
NICKLE
LEAD Te 3.00
TING TTM 947

LEAF

NOT SIGNIFICANT
EMEAN =CHE AN

NOT SIGNIFICANF

NOT SIGNIFICANT
EMEAN=CMEAN
SEGNIFICANT
50
NOT SIGNIFICANT
EMEAN =CHE AN
EME AN SCHE AN
SIGNIFICANT
NOT SIGNIFICANE
SIGNIFICANT
125
NUT SIGNIFICANT
EMEAN=CNEAN
EMEAN =CMt AN
E WEAN =CKE AN
NOT SIGNIFICANT
SIGNIFICANT

EXPERIMENT WO=1

EME AN-CHEAN®

EME AN-CHE Ae
EME AN-CHE AN =

EMEAN-CHEAN=
EME AN-CME AN=
EME AN-CME AN=
EWE AN-CME AN =
EME AN-CME AN ®

EME AN-CME AN=

EMEAW-CME AN *
EME AN-CME AN®

-.80

A585
“4,50

73.35
=10
74.05
-2.00
3.95
~.20

1.50
2.55

OEG= 1
DEG= 1
OrG= -1
DEG= 1
OEG= 2
vEu=

DEv=

DEG=

DEGS 2
LtG= 2
bEG= 2

EMEAN=
EMEAN=
EMEAN®
EMEAN=
EMEAN=
EMEAN=

EMEAN=

EMEANE
EMEAn =
EMEANE
EMEANS
EMEGN=
ExEAN®

270
1.70
4.45
00
+25
8.15

1.05
1.70
4645
3645
4.50
7,60

95
1.70
4.45

+00
5.50

10.25

EDEV=
EDEV=
ebEVe
EDEV=
EDEVe
EDEV=

EDEV=
EDEV=
EDEV=

eDEV=

EDEV=

EDEV=

EDEV=
tDEV=
EDEV=
evEV=

EDEV=

EDEV=

214
014

CMEAN®
CMEAN=

200 “CHE ANS

200
00
o2)

07
«07
207

207

71

42

207
07
07
07

re)

35

CHEAN=
CMEAN®
CHEANS

CMEAN=
CMEAN®
CHEAN=
CMEAN=
CHEAN®
CHEAN®

CMEAN®
CMEAN=
CHEAN®
CHEAN®
CMEAN®
CHEAN®

1,50 COEVTM
1. 70CDEV=
9.00 CDEV=
4.50 COEVe

+25CDEV=
21.50 CDEV=

1.15 COEV=
1.70CDEV=
4 ASCDEVE
7.50 CDEV=
6.50 COEV=
11.75 COEVs

1.15 COEVs
1.70CDEV=
4 .G5COEVs
sOOCOEV=
4.00 COEVs
7.70 CDEV=

200
00
200

235

00
14


ican

OYSTER

CADN TUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADN UM
CHROMIUR
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM LUM
CHROMIUM
CUPPER
NICKLE
LEAD
ZIG

CADMIUM
CHRUMIUM
COPPER
NICKLE
LEAD
ZINC

CADH TUN
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUR
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

INTERNAL EXPERT
DAY NUMBER 0
NO EXPERMENT MEANS 1.77
NO EXPERMENT HEAN= 10.40

NO EXPERMENT
NO EXPERMENT
NO EXPERMENT
NO EXPERMENT

DAY NUMBER
Ts 11.50
Ts 2450
Ts 7.82
Ts 3.15
Ts 3.08
Ts 13.12
DAY NUMBER
T= 8.50
Ts 1.58
Ts 4.78
Ts 1.25
Te +50
T= 2.21
DAY NUMBER
Ts 3.77
Ts 2.39
Te 8.51
Te 2614
T= 2.92
Ts 2.70
DAY NUMBER
Ts 1.31
Te 2659
T= 10.30
Ts 6642
Ts 3.05
Te 2-44
DAY NUNBER
Ts 00

+00

+00

+00
= +00
T= 206
DAY NUMBER
T= 3.66
Te 1.63
Ts 1.46
Ts 1.64
Ts 2.74
Ts 5.46

HEAN® 60
MEAN= 19.23
MEAN® 7.23

MEANS 657.37

1
SIGNIFICANT
NOT SIGNIFICANT EMEAN-CHE ANS
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

5

SIGNIFICANT

NOT

SIGNIFICANT

SIGNIFICANT

NOT
NOT
NCT

25

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

SIGNIFICANT

NcT

SIGNIFICANT

SIGNIFICANT

NOT

SIGNIFICANT

SIGNIFICANT

NOT

59
aot
wot
NOT
NOT
wet
WoT

BS
net
NOT
wot
NOT
NOT
wot
125
WOT
Not
NOT
NOT
NOT
NOT

SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICAKT
STONIFICANT
SIGNIFICANT

MENT NOe2

DEVIATION®=
DEVIATION=
DEVIATFION=
OEVIATION=
DEVIAT luke
OEVIATION=

EME AN-CME AN®

EMEAN-CME AN®
EMEAN-CME AN&
EME AW-CHE AN®
EMEAN-CME AN®

EMEAN-CHE AN®
EME AN-CME AN®
EME AN~CME AN ®
EME AN-CME AN
EMEAN~CME AN®
EMEAN-CMEAN=

EME A-CME AN®
EME AN-CMEAN®
EME AN-CME AN®
EME AN-CMEAN=
EME AN-CHE AW
EME AN-CME AN®

EME AU-CME AN®
EME Aw-CME AN®
EME AN-CME AN =
EME AN-CME AN=
EME AN-CME AN =
EME AN-CME an =

EME AN-CHE AN®
EME AN-CME AN=
EMEAN-CME AN=
EME AW-CHE ANS
EME AN-CME AN®
EME AN-CME AN=

EME AN-CMEAN®
EME AN-CME AN®
EMEAN-CMEAN=
EME AN-ChE AN=
EME AN-CME AN®
EMEAN-CME AN=

206
53
3.00
1.53
3614
145.94

2.10
6.53
72.97

712.93

4.90

425.50

95
2065
47.90
3.00
260
112.50

0
172.17

1.25
13,00
90.05

9.45
5.55
132.96

4.60
11.40
421.50
14.00
16.70
1651.70

095
11.15
91.90

8.50
9025
637.10

NUK®
NU
KUM
NUM®
NUM=
NUM®

vEGs
DEGe
DEG=
VEG=
vEG=
DEG=

DEG
DEG=
DEGs
DEG=
DEGs
OEGs

DEG=
VEG=
DEGs
OEus
DEG=
OEG=

DEG
vEGs
DEG
DEG=
DEG=
GEG=

DEG=
DEGs
DEGs
GEGe
DEG
veoe

DEG=
DEG=
Deus
DEG=
DEG=
DEG=

weovenw

BREE ES NUNN NK PEE EEE

COCCOO Hee Eee

EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN=
EMEAN=

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN®

EMEAN®
EMEAN®
EMEANS
EMEAN=
EMEAN=
EMEAN®

EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN=
EMEAN=
EMEAN=
EMEAN]
EMEANS
EMEAN®

EMEAN®
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS

4.90
5290
158.77
12.30
12047
920.80

4.65
12.95

9.70
642.70

5.90
21.23
223.60
2 3
14.63
884.77
3.80
29.80
214.20
25.90

10.90
842.40

7.40
18.40
720.80
25.00
23.70
2386.90

3.45
28.35
336.40
25.40
14.35
1028.60

EDEVe
EDEV=
EDEV=
EDEVe
EDEV=
EDEVs

EDEVs
EDEVe
EDEV=
EDEVs
EDEV=
EDEV=

EDEV=
EDEVs
eDEV=
EDEV=
€DEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEVs
EDEV=

EDEV=
EDEVe
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEVe

CHEAN=
CHEAN®
CHEAN=

CHEAN®

CHEAN®
CHEAN=
CMEANS
CHEAN=
CHEAN=
CMEAN=

CMEAN=
CHEAN=
CHEAN®
CHEAN=
CMEAN=
98.48 CHEAN=

200 CHEAN=
200 CHEAN=
200 CHEAN=
+00 CMEAN=
+00 CHEANS
030 CHEAN=

+00 CHEAN=
+00 CnEAN=
+00 CHEAN=
200 CHEAN=
200 CMEAN=
+00 CHEAN=

+21 CHEAN=
5.59 CHEAN=
51.34 CHEANS
4.24 CMEAN®
2.76 CMEAN=
95.32 CMEAN=

32

2.60
12.43
85.80
25223

7.57

495230

3-70
10.30
99.20
19.00

9.10

530.20

4.00
15.30
146.40
1 7
8.33
712.60

2.55
16.80
124.15
16.45
5.35
709.50

2.80
7.00
299.30
11,00
7.00
735.20

2.50
17.20
244.50
16.90
5.10
391.50

CDEVe
CDEV=
COEVe
CDEV=
CDEV=
CDEV=

COEVe
COEV=
CDEVe
CDEV=
COEVe
CDEV=

CDEV=
COEV=
COEV=
CDEV=
COEV=
COEV=

CDEV=
COEV=
COEV=
CDEV=
COEV=
COEVe

COEV=
CDEV=
COEV=
COEV=
COEV=
CDEV=

cDEV=
CDEV=
COEV=
coevs
CDEV=
CDEVe

200


DISCLAIMER

This report has been reviewed by the Narragansett
Environmental Research Laboratory, U.S. Environmental
Protection Agency, and approved for publication. Approval
does not signify that the contents necessarily relfect the
views and policies of the Environmental Protection Agency,
nor does mention of trade names or commerical products
constitute endorsement or recommendation for use.

ii


oot

MANGROVE ROOT
DAY NUMBER = 5.
CADMIUM T= 3-00 NOT SIGNIFICANT
CHROKIUM EMEANSCKEAN
COPPER EMEAN =CKE AN
NICKLE = T® +00 NOT SIGNIFICANT
LEAD Ts 3.00 NOT SIGNIFICANT
ZINC Ts 3.77 NOT SIGNIFICAWT
DAY NUMBER 50
CACMIUM Ts 3.00 NOT SIGNIFICANT
CHROMIUM
COPPER EMEAN=CMEAN
NICKLE T= aan SIGNIFI CART
LEAD Ts 2.83 NOT SIGNIFICANT
ZINC Ts 12.63 SIGNIFICANT
DAY NUMBER 125
CADKIUK T= +28 NOT SIGNIFICANT
CHROMIUM T= 3.02 NOT SIGNIFICANT
COPPER = T= +00 NLT SIGNIFICANT
NICKLE | T= 5.28 SIGNIFICANT
Leap 1 3.06 MOT SIGNIFICANT
ZINC Ts 4.33) SIGNIFICANT

EXPERIMENT NO=1

FMF AR=CME AN=

EME AN-CME AN=
EME AN-CME GN =
EME AW-CHE AN =

EME AN-CME AN

EME AN-CME AN=
EME AN-CME AN=
EME AN-CME AN ©

EME AN-CHE hiv
EME AN-CME AN =
EMEAN-CHE ANS
EAE AN-CME ANS
EME AN-CHE AN =
EME AN-CME ANS

2.00
12.65

nmol2
3.77

DEG=

DEG=
DEG=
DEG=

GEG=
DEG=
DEG=
DEG=

DEG=
DEG

QO vtG=

5.75
3.00
19.05

DEG=
DEG=
DEG=

ny NNO

BUUWWe BAH

EMEAN=
EMEAN=
EMEAN®
ENEAN=
EMEAN=
EMEAN=

ERMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN®

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=

1.20
1.70
4645
00
4.00
11.55

1.80
1.78
4.45
9.00
7250
23.95

16035
7.00
4.45
16.45
10.50
49.35

EDEV=
EDEV=
BOEV=

EDEV=

EDEV=

EDEV=

EDEV=
EDEV=
EDEV=

EDEV=

EDEV=

EDEV=

EDEV=
EDEV:

EDEV=
eDEv=
EDEV=
EDEVe

-00 CHEAN®
200 CMEAN=
00 EHEANE
200 CHEAN=
«00 CMEAN=
v34—CHEAN®

+00 CNEAN®
100 CHEAN=
200 CHEAN=
CMEAN>
CHEAN=
CHEAN®

CMEANS
CMEAN®
00 CMEAN=
2005 CHEAN=
o71 CHEAN=
6.29 CMEAN=

9

1.05 CDEV=
1.70CDEV=
4 GSCOEV=
6.00 COtvs
5.50 CDEV=
7.65 COEV=

1.80 COEV=
1.70CDEV=
4.45CDEV=
5.50 COEVe
5-50 COEV=
11.30 CDEVe

1.17 COEV=
3.23 COEV=
4.45 CDEVe
10.70 CDEV=
7.50 COEV=
30.30 CDEV=

1.41

1.34
200
17

1.22

3.87


30KM
—s

MONA PASS

“18°N 67°W

Figure 1. Study site location.

21a


DONO TOO DLDNITQCANONNOMDOION TSA MNADOTTRRONNONNMONINO ATS

NOOMANNDAUVANOOONOAMENONNONOCOBARTOTVOIONTV ONO LON
cower ece ee eee eee eee wee me ee ee ow eee ee ee eee coe ee eee
OTM TNNAR STH TT OMOTRAOEPNHTOO.OR PNATRANGARAR TONS
AMFMAMOMMAAMAMM MNT THNT Sy tert dddaaa tte

TOONNDMONDON

cee eee ee eee

oat vt z * La

eee eee °
RAM ANNATNTANOMMHGNDBOATTONMOOMINO DO rPOUNNNDDO
AN ttt tt tt ot ot At ttt tet tet

Ce ee ee eS Oe ee

REQOOHMQOCOMAANQOAONANHALE YER SQTOVTANDOHRENMTONANOVNOR aan

eee ee eee ree eae re ee ee ee Pr ere ere rr ee ee ae ee ee ee

DQOAEKE EOF OME oO BARA RAR SS SSR GOR SHARAN ANNANA ANAM MORAN

«7
AMT OOMNANTOO# DOME TOM DDOWONMAANONOCOHRXDOMONN TO TOHROANE HAO
Pr re ee ee re ar re ee D Carer area are ae ar
OPER oornrr BONN COR SN AORANGOS GMOUNMANNNARANM OTE TATA
* a
2
2
< &
~
SOGANAOOARNDODMAOTONADQCOONOOR aR NHTADIORENHE TONE NHONT ORE
Qeeee Cee ere er er ere ee rs ee eS cee eee i eer ere re Se eed oe oe eee ee
ANNAN NNN A NAN NO NNN NNN ANNAN ttt tt ob tet
° * *
w a 2
ra 2 2
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Oo
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HM HNSDOO
ANNA NOAAA

85

aman am all
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125
125
125
125
125
125

NNN

FAABHODORAHODOH HHO OORH AH OOOR NA DOO MA aH HHOOO HA HOOONAMAMOOORM

NNNNNANNNANNAN NNN NNN NNN NNN NN NNN NNN NNN NNN NNN

DAMA HHNNNNMNNDDUNDMNADYNNUDYNNMNNNANNHNNNYNNHNDAAMaANHDHAMNHOOMNMRADNODHA

BSVVEVVVVVVEVYVYYYEYYVYYEYYVYEYYUYYYUYYUYUTIIITCICITITITCLTCrTrrrTzrrrrir.e

80


YUN TIRTOTOOON ONE Trek wer

wy 2

: eee ee er er ee er er ee)
PHADIOFAAAOKANNOMNATCHAKHOOANTAM
AA AN tt tit rN ENR. SOMMOON IN

BAnAnr oonr nr oon

ea ar eer are ee
MAMONMNDOODONMINAND
tana an ma tt

ARAVA TRARANV ARM ANTS KORSAMNOCCORAHNGDSCHONANR ME DCDLL

eee eee . . . ° a eer a a re ee

QANVTTOEMMON TT OSS a one momen Ar TNNOONH AA

ANd ae HM AMAMAMMAMMA MA mam MMODONQQORN
aw =

TINGS SHOAOAMARWOGIIIRHYN AIGA IOORNWAOHMAOMNTGS
Ce aa ere ret eee eee Oe ee ee err ee reat at ere eres
BRE NMOR ORR OMOR RR OP ORR OR DORR ONE RORR TT ONNOWD
QANANnOnvoed NHOOTONn OM
DONNA AM 6
.
ALM OAMNANAMONTFON NOOO TVONONONMNHRONNNOTANR AR
Cee ee ee ear re ar re er ee ere ee aD
AMMNANATM ATMS TST TT TOMT TOTS NOMENHEAN BANOO
Ror orteer
aa a

SORTED DATA

MN OOCOFANNNNOOCOO RAMAN NDKADNN NNN ANHKHHNHO
aw NN ANNAN NNN
aa

COCO HAHOHOHORHOOHAOOHHODDOHAOOOCO MHOC RH OH ORG
EE tet tt
DO eR Se ER NANYNNNNNHHNHNDHNHHNHNHNHNNHDDOOBAGOO

ZzZocgooccos,9coOGeoomTeCcCGCCOCCoOOGCFOOAFoOOoOaaaaaoa0a

75


SECTION 2
METHODS

Construction of Sea Water Tanks

The sea water system and tanks were located near Bramadero
Bay, approximately 8 km from Mayaguez, Puerto Rico (Fig. 1).
Two tanks (9.2 mx 0.9 m x 1.1 m) were constructed from 1.9 cm
exterior grade plywood with 5 cm x 10 cm external framing (see
Fig. 2). These tanks were sealed with two coats of fibreglass
epoxy resin and leached in running sea water for 2 weeks. The
tanks were located in an open structure. The roof of the structure
was fabricated of galvanized corrugated roofing. The underside
of the roof was covered with plastic sheeting to prevent zine
contamination.

Flowing Sea Water System

Each tank was filled with 4.2 m? of calcareous beach sand

(7% CaCO.) from Aguada, Puerto Rico. The mean depth of sand“was
0.5 m with an additional 0.1m of sand over the last 3 m of tank.
The tanks were then filled with sea water to a mean depth of 0.5 m
and flushed for an additional week.

The sea water for the system was pumped from the dock of the
Center for Energy and Environment Research at a depth of 3 meters.
The pump was a Sears Dynaglas 1 hp (230 volt). A plastic minnow
trap was used as a course prefilter for a Filterchem ® combination
filter and foot valve at the pump inlet. All piping used was
grey schedule 40 P.V.C. The water traversed 73 m to a charcoal,
sand and gravel filter (0.5 x 0.5 x 1.0 m), then to a 1000 liter
settling tank. The settling tank was constructed_of 1.9 cm

thick plywood lined with two coats of fibreglass R and epoxy resin.
The water was then gravity fed to the two sea water tanks with

the flow split using P.V.C. "T" joints. The flow rate,in each
tank was monitored and maintained at 8.2 to 8.7 4g min ~ by PVC
ballvalves. The total water volume was 3.1 x 10° liters. The
turnover time was 5.9 hrs per tank. The inflowing water was
diverted below the surface by submerging the inflow end using

a perforated inflow pipe (Fig. 2). The sea water was continuously
aerated with an air pump and air stones.


ZN THALASS (A LCLEAVES— EXP. 2

s |
= 4
xz
Qe 4
~
“A 4
Y a 4
rm)
u 4
zd
z=
WW 4
=
-e1
Lae
é g Fy
pRYS
ZN URCHIN GUT- EXP. 2
“ew 4
= 4
x
a a4
S
~ 4
Y ae 4
uw
wl 4
z m4
G 4
=
a 4
EEL nn
s 6 g rr
DAYS

Figure 25. Mean net uptake of Zn between experimental and control,
Thalassia leaves and urchin guts ~ Experiment II.

45


HARA AAAS FH A AHHH HUUHHHUHUDUHNHUHNHUHUH HUY

Cel al al ell ll lol ll ol ll

RN NNR NARRNRNRNNNNNURNNNNNRNNNNN NNN NNNNNNNYNNNNYNNNNNDND

OP RP HOC OR PH OOP HH OOOH HOO OMB EEE HOO OPK HOO OH RHR OOOKRHOOORRF

SORTED
5
5
25
25
25
25
25
25
50
50
50
50
50
50
85
85
85
85
85
85
125
125
125
125
125

ra
N
w

UNUM eee RR OOSo

3

ee eee

PAINT R ORM UNOTWRKWOVDONWOUURHOKN

rr ar or er ee ar er rr rs

WVNUNNOUUBWWEDNVVE NEN EH HEE NEE

A ek ee ee RO)

rer ere er ree Serer er eer re aes

.

SIU PTR WUYYUR VOUT TUR OY

FPP OL
Hrwo

+
w
~

54.
54.2
52.0
54.0
50.2
51.8
16.6
16.3
15.5
16.4
15.6
14.2
12.4
13.9
51.0
49.6
59.3
37.5
41.0
39.3
32.2
31.8
37.5
37.1
37.5
35.4
38.8
44.4
57.4
58.5
51.9
116.9

@

87

-

COOK ON NTT NMP ®MYVIVIV SIMPY NROMMOWOCMO@®OUA
ee ee

DUARHNUATLU LK DOENBOEAHAFWHROTOBBRONRNBRTOWONOROO

PRE E ee
©
ew arr ree ee ey

ee
ww
ee

15.0
15.3
15.4
19.7
20.6
22.4
22.8
22.5
26.9
30.4
34.5
24.8
24.9
25.0
29.3

rire arn ara ary

CORP KPOMRODMYM®YVINSPYNMDOMINRUSE SD

NUP NOH DBDON FRE WHE UTI O OOH YUE UO

Peer

wee
ooo

28.1
29.4
24.0
24.6
24.0
21.2
20.2
55.2
55.4
59.3
40.9
42.9
42.9
28.6
30.4
42.0
41.5
41.7
23.5
27.5
30.3
44.1
44.2
44.4
73.6

er rhe rr er ar rr ar ar ar ar er ar Or Or Or Or re rer a ne ary

WODOWNOADR ERE E UTES HEP WOR UE SOY DH OUWOHKUWORYNONY

RreeeE

Pree
AOMDOUENATNENOOONNANNOCMYURORIVFYVHPHRUNTNUNFE TRO UU

oer eee ee were

14.8
13.9
15.3
14.5
13.7
15.5
15.3
16.9
16.0
15.2
18.6
14.1
10.9
ll.é
15.4
15.1
14.9
16.3
15.5
14.6
18.2
17.4
17.0
16.1
15.0
15.2
28.3
26.6
28.0
25.2
26.9
2544
28.0
24.4
34.9
40.1
37.0
39.3
36.9
33.4
23.6
22.8
33.0
32.6
30.6
41.8
45.7
53.7
39.2
41.4
40.5
31.3


Montgomery, J.R. and J.E. Echevarria. 1975. Organically complexed
copper, zinc, and chelating agents in the rivers of Western
Puerto Rico. In: Mineral cycling in Southeastern Ecosystems.
E.G. Howell, J.B. Gentry and M.H. Smith (EDS.). ERDA
Symposium Series (Conf-740513) p. 423-434.

Nair, K.V., M.C. Balani, and S.G, Shringarpure. 1973. zn®° up-
take By laboratory cultures of Microcystis littoralis and
Chlorella vulgaris. J. Mar. Biol. Assoc. of India 15(1):
302-305. ~

Odum, H.T., P.R. Burkholder, and J. Rivero, 1959. Measurements
of productivity of turtle grass flats, reefs, and the Bahia
Fosforescente of Southern Puerto Rico. Publ. Instr. Mar.
Sci., Univ. Tex. 6: 169-170.

Phelps, D.K., G. Telek, R.L. Lapan Jr. 1975. Assessment of
heavy metal distribution within the food web. In: Marine
Pollution and marine waste disposal: International Congress,
2nd, San Remo Dec. 17-21, 1973. Pearson & Frangipane,
Pergamon Press, Oxford and New York.

Salotto, B.V. and J.B. Farrell. 1971. Preliminary report - The
impact of sludge incineration on air and land. Prepared by
Advanced Waste Treatment Research Laboratory, NERC,

Cincinnatti EPA task force on sludge incineration, July 12, 1971.

Schroeder, P.B. 1975. Thermal stress in Thalassia testudinum.
Ph.D. dissertation, University of Miami, Coral Gables, Florida.

Shuster, C.N. Jr., and B.H. Pringle. 1969. Trace metal ac-
cumulation by the American eastern oyster, Crassostrea
virginica. Proc. National Shellfisheries Assoc. 59: 91-103.

Silviera, D.J. and L.E. Sommers. 1977. Extractability of copper,
zinc, cadmium, and lead in soils incubated with sewage sludge.
J. Environ. Qual. 6(1): 47-52.

Street, J.J., W.L. Lindsay, and B.R. Sabey. 1977. Solubility
and plant uptake of cadmium in soils amended with cadmium
and sewage sludge. J. Environ. Qual. 6(1): 72-77.

Strickland, J.D.H. and T.R. Parsons. 1965. A manual of sea water
analysis. Fish. Res. Bd. Canada Bull. #125. 203 pp.

Turner, R.E., J.W. Day Jr., M. Meo, P.M. Payonk, J.H. Stone,
T.B. Ford and W.G. Smith. 1976. Aspects of land-treated
waste applications in Louisiana wetlands. In: Proc. of
the Natl. Symp. on Freshwater wetlands and Sewage Effluent
Disposal Ann Arbor, Michigan. D.L. Tilton, R.H. Kadlec,
C.J. Richardson (Eds.). p. 147-167.

66


CD FOULING ORGANI SMS— &xP 2

s
= 7 4
2 .s |
s 2 7
A
Co.
wont |
L J
-.2]
Zz 4
Cc -.s4
ui 4
Zz -.7]
-.s4
4 La T i —T T +r T 1
s Es a
Ey
CRYS
<p SEWRGE- ©xP 2
zen
zse
= 220
i
a
12
z
&
Y ee
=e
ea + =
—r r —+ 1
. & s x
8
cCRYS

Mean net uptake of Cd between experimental and

Figure 7.
control, fouling organisms and sewage - Experiment II.

27


CR THALASSJA CEAVES—- xP 2

=o |
oa
cs
x |
~
Yue |
la
¥
z |
Cc
we |
a
:
é g z
CRYS
<cR. URCHIN GUT—- EXP. 2
so 4
-
b we 4
t |
~ se]
Vv
yd
Wo2nd
y
zie]
yo |
2 |
= é g Fy
CRYS

Figure 19. Mean net uptake of Cr between experimental and control,
Thalassia leaves and urchin guts - Experiment II.

39


80T

NO EXPERMENT MEAN
NO EXPERMENT MEAN=
NO EXPERMENT MEAN®
NG EXPERMENT MEAN=
NG EXPERMENT MEAN=
NO EXPERMENT HEAN=

THALASSL
DAY NUMBER
ADH IUK
CHROMIUN
COPPER
NICKLE
LEAD
ZINC
OAY NUMBER
CADMIUH T= 67
CHROMIUM T= 1.30
COPPER «T= 6.57
NICKLE = T= 7.61
LEAD Te 11671
ZINC Ts 1.02
DAY NUMBER
CAOMTUK T= 1.75
CHROMIUM T= 0.04
COPPER Te +73
NICKLE | TR 1.81
LEAD Ts 4.80
ZINC Ts 1.21
DAY NUMBER
CADMIUM T= +33
CHROMIUM T= 3.18
COPPER = T# 2.79
NICKLE T. 4.64
LEAD Ts 206
ZINC Ts 1.43
DAY NUMBER
CADMIUM Ts 34
CHRON HUH T= 3.53
COPPER T= 5.68
NICKLE = T= 4.42
Lead Ts 8.36
INC Ts 1.69

RooT

0

1
NGT SIGNIFICANT
NGT SIGNIFICANT
SIGHIF ICANT
SIGNIFICANT
SIGWIF ICANT
NOT SIGNIFICANT
5s

NOT SIGNIFICANT
NOT SIGNIFICANT
wGT SIGNIFICART
NOT SIGNIFICANT
SIGNIFICANT
NCT SIGNIFICANT
25
wCT SIGNIFICAKT
SIGiWIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
50
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT

EXPERIMENT NOD=1

215 DEVIATIUAE
5.00 DEVIATION=
ih DEVIATION®

+45 DEVIATIUN=
15.50 DEVIATION=

25.40 DEVIATIUN=

EME AN-CME AN=
EME AW-CMEAN®
EME An-CME AN =
EME Aw-CME ANS
EME An-CHE all s
EME AN-CMi AN=

EME AN-CHE ANS
EMEAN-CME AN
EME Aiv-CHE AN =
EME AW-CMe Aly=
EME AN-CME AN =
EME AN~CME AN=

EME AW-CME AN=
EME AN-CME AN=
ENE Ad-Cité Als
EME AN-CMEAN=
EME AW-CME ANS
EME AN-CKE AN=

EME AN-CHE AN=
EMEAN@CMt AW=
EME AN-CME AN®=
EME AN-CME AN=
EME AN-CME ANS
EME AN-CHE AN =

+00 NUK
1.27 NUMe
+00 NUH
+00 NUH
+00 NUM=
2.55 NUM=

7284

DEG:
DEG=
DEG=
vEG=
DEG=
DEUS

DEGs
DEG
DEG=
OEG=
DEG=
DEG=

DEG=
DEG=
VEG=
DEG=
DEG=
DEG

DEG=
DEG=
OEG=
DEG=
DEG=
OEG=

NRNNNN

pepese

PREERD SeOUree caoore

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEANS
EMEANS
EMEAN=
EMEAN=
EMEANS
EMEANS

EMEANS
EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=

EMEANS
EMEAN=
EMEAN=
EMEANS
EMEANS
EMEANS

1.15
26.15
15.65
43.50
28.65
34.15

1.84
32.83
14.10
46645
27,30
34.62

1.67
40.02
19.12
45.95
22.10
34,37

2.40
54.32

EDEV=
EOEV=
tDEV=
EDEV=
EDEV=
EDEV=

EDEV=
cDEV=
tDEV=
EDEV=
EDEV=
EDEV=

50.37

44.45. EDEV=

37.15

EDEV=

221
1.77
221
3.66
35
1.63

1.29

CmMEAN=
CMEANS
CMEAN=
CMEAN=
CMEAN=
CMEAN=

CMEAN=
CMEANS
CMEAN=
CHEAN=
CHEAN=
CHEAN®

CMEAN=
CMEAN=
CMEAN=
CMEAN=
CMEAN=
CHEAN®

CMEAN®
CHEAN=
CHEAN=
CMEAN=
CHEAN=
CMEANS

18

1699
21.15
12.37
29.57
20.77
31.95

3.00
29.83
14.77
42.67
17.85
36.85

1.65
27.40
13.30
34.23
22.22
28.40

2.30
26.65
13.95
34.35
12.20
27.65

COEV=
COEV=
CDEV=
COEV=
COEV=
COEV=

COEVE
COEV=
COEV=
CDEV=
COEV=

COEV=
COEV=
COEV=
COEV=
CDEV=
CDEV=

COEV=
COEVe
COEV=
COEv=
COEV®
COEVE


ZN THALASS!A LERVES— EXP. 1

ae |
=
£ |
~ sp J
2
VV
2 |
ld
ww 1@ 4
Zz
Cc 4
lu
|
- a
8 ®
CAYS
ZN URCHIN GUT- EXP. i
ee
4
= ce |
oa
S 4
S +e 4
1
4 4
w
ae |
Zz
Cc
= |
=
@ 4
—— 4

Figure 24. Mean net uptake of Zn between experimental and control,
Thalassia leaves and urchin guts - Experiment I.

44


6eT

URCHIN

CADMIUM
CHROMIUN
COPPER
NICKLE
LEAD
mac

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
LINC

CADHIUN
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHRUMIUM
COPPER
NICKLE
LEAO
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
TING

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

INTERNAL EXPERIMENT NO#2
DAY NUMBER o
NO EXPERMENT MEAN= 2.17 DEVIATION=
NO EXPERMENT MEAN= 7.37 DEVIATION=
NO EXPERMENT HEAN= 4uo5 DEVEAT Hine

NO EXPERMENT MEAN= 16.03
NO EXPERMENT MEAN= 18.47
NO EXPERHENT MEAN= 27.33

DAY NUMBER
Te 1.41
Ts 7.77
Ts 3.17
T= 2.05
T= 1.51
DAY NUMBER
Ts 7.78
T= 5.00
Ts 8.46
T= 9.00
T= 5.06
Ts 4.01
DAY NUMBER
T= 4.95
Ts 6.79
Te Felt
Te 10.53
Te 4.49
Ts 8.05
DAY NUMBER
Te 1.00
Ts 2.40
Ts 2.71
Ts 5.13
Ts 229
T= 1.62
DAY NUMBER
Ts 5.00
Ts 53.67
tT: 9.60
Ts 25.94
Ts 7.06
Ts 1.863
DAY NUMBER
Ts 447
tT 12.05
Ts 26.77
tT 2.38
Ts 11.57
Ts 6.46

a
NOT SIGNIFICANT
SIGNIFICANT
EHEAN=CMEAN
SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
5
SIGNIFICANT
SIGWIF ICANT
SIGNIFICANT
SIGKIF ICANT
SIGNIFICANT
SIGNIFICANT
25

SEGRIF ICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
50

NGT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
85

SIGHIF ICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
125
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

DEVIATIUN=
OEVIATION=
DEVIATION=

EME AN-CMEAN=
EMEAN-CHEAN=

EMEAN-CME AN®
EMEAN-CME AN®
EME AN-CMEAN®

EME AN-CME AN®
EME AN-CHE AN®
EME AN-CME AN =
EMEAN-CME AN=
EME AN~CHE AN =
EMEAN-CHE AN®

EME AN-CME AN=
EME AN-CMEAN®
EME AN-CME AN =
EME AN-CHE AN®
EMEAN-CME AN®
EME AN-CHE AN =

EME AN-CME AN®
EME AN-CME SN&
EMEAN-CME AN =
EME AW-CHE AN®
EME AN-CME AN=
EME AN-CHE AN®

EME AN-CME ANS
EME AN-CME AN=
EME AN-CME AN =
EME AN-CMEAN®
EMEAN-CME AN®
EME AN-CME AN=

EME AN-CME AN=
EME AN-CMEAN®
EME AN@CHE
EME AN-CME AN®
EME AN-CME ANS
EME AN-CME AN®

206 NUM®

1 NUM®

200 NUM®
245 NUKE
1.51 NUM®
2.79 NUMe

-.07
93

1.23
2.40
2.37

-.37
3.80
2.03
7.46
4.73
7.90

7023
20.93
3.87
29.50
4040
10.20

03
71.36
--70
-3.20
7220
4.40

17
42.33
4.97
25.90
5.57
+73

«33
10.63
10.97
3.83
23.80
18.67

DEG=
DEG=

DEGs
OEGs
OEG=

DEG=
DEG=
DEG=
DEG=
DEG=
DEG=

DEG=
DEG=
DEG
DEG=
DEGe
DEG=

DEG
DEG=
DEG=
DEG=
OEG=
DEG=

DEG
DEG=
DEG=
DEG
DEG=
DEG=

DEG
DEG=
CEG=
DEG=
DEGs
DEG=

veowuw

PRPEDS FEE PD

PEDDPD PEPKDS SERRE

Spee ne

EMEAN®
EMEAN=
EMEAN®
EMEAN=
EMEAN®
EMEAN=

EMEAN®
EMEANS=
EMEAN®
EMEAN®
EMEANS
EMEAN®

EHEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN]

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEANS
EMEAN=
EMEANS
ENEANS
EMEAN=
EMEAN=

EMEANS
EMEANS
EMEANS
EMEAN=
EMEANS
EMEANS

1.97
6.93
445
20.10
20.07
24.43

1.77
18.70
411.33
39.17
18.17
43.20

1.43,
36.33
15.07
56.83
26.37
41.40

2.20
13.90
411.00
16.23
23.47
27.37

2.37
70.77
15.47
47.50
20.30
42.53

2.37
28.27
20.80
34.53
46.90
54.70

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
€DEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
€OEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
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EOEV=
EDEV=
EDEVe

EOEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEVE
EDEV=
EDEV=
EDEV=
EDEV=

206
206
06

1.80
2.62

06
1.31

1.19
225
3.21

206
2.95

4.84
84
2.05

00
260
44

1.00
3.13

06

CHEAN=
CMHEAN=
CHEAN=
CHEAN=
CHEAN=
CHEAN=

CHEAN=
CMEAN=
CHEAN=
CHEAN=
CHEAN®
CHEAN®

CHEAN®
CMEAN=
CHEAN=
CMEAN>
CMEAN=
CHEAN=

CMEAN=
CHEANE
CHEAN®
CNEAN=
CHEANS
CMEAN=

CHEAN=
CMEAN=
CMEAN=
CHEAN=
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CMEAN=

CMEAN=
eM EANS

“EAN
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CHEANS
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39

2.03
6.00

COEV=
COEV=

4.45C0EV=

18.67
17.67
26.80

2.13
14.90
9.30
31.77
13.43
35.30

1.67
17.40
11.20
29.33
21.97
31.20

2.17
15.20
11.70
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23.67
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2220
28.43
10.50
21.60
14.73
41.80

2.03
17.63
9.83
30.70
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COEVe
CDEV=
COEV=

CDEV=
COEV=
CDEVe
COEV=
CDEV=
CDEV=

CDEV=
CDEV=
COEV=
CDEV=
CDEV=
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COEV=
CDEV=
CDEV=
COEV=
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CDEV=

CDEV=
COEV=
COEV=
CDEV=
CDEV=
COEV=

COEV=
COEV=
COEVe
COEV=
COEV=
COEV=

+06
+20

267
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70

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lo
217
78
1.60
1.15

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445
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1.47
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10
275

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00
1.29
87
1.46
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12
206
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290
2.14
3.07


PB THALASS (A CEAVES—- EXP. i}

4
a |
= 4
i
sa
S
tO]
Y ue J
rit]
v
z =]
ca
ui 4
z=
LJ 4
T , ——+ t r 1
. s g Fa
DAYS
PB URCHIN GUT-— EXP 1
7
3e@ 4
=
a. 4
a
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2a J
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Y 4
i]
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Zz
Cc 4
wi
= a 4
T ——1—+ T a

az

DAYS

Figure 20. Mean net uptake of Pb between experimental and control,
Thalassia leaves and urchin guts - Experiment I.

40


6 = Concentration (ig/g dry weight) Cd
7 = " " cr
8 = " " cu
gz " " Ni
10 = " " Pb
11 = " " Zn

Multiple values for the same sample represent replicate
samples. The value -999.9 is an indication of a value that could
not be detected. All manipulation of the data used one half
minimum detectable concentration (MDC) when a value was less
than or equal to the MDC. The MDC values are listed in the
Results (Table 4).

69


ee ee ewe eee ar ee ar er ree rr

be See Bet

° ca

PS COHA SORBET FARHAD OORMAEMMAMMODOSANROHSHNHANEIAS
Att tte

BETBDONMGDDGCOTONATMO DVN TDVODDAAN AAA THHH TT ERO ANVOOR sear

Le Te Oe ST Oe

MMOMNN AAA AMTN AMT TNM TET TTT TIMER OF ONOMMNTTMMOAEPNHOEO

AN MRONONMANNNVOMOGTNADNOQOOOH OCONMOrHMTOOTEMNHOSNHONHNHETY

ee eee oe ee ee ee eee

SORTED DATA
464
4.7

25
25
50

DODOCO KAA MAMAN NNNHNNAMNNNNNNOCOCDOOOHNMNNNHH
wn NANNANNAN ADH AN AN ODD DOD

125
125

ARMOR O MMM MMOOCOHAHODCO HAA OOOHAHAHOOCO MAAN OOO NH ARNOOOH AMAA OOO
AMM MAMAN NNN NN NNN NN NNN NNN NNN
CO et ee eee er kkk)

PINNNNN VE VV VV EVEYVYE EE YYVYVYEYYVYVUYYUEVEYEYUYVYEYUYEYVYUUYUYUUUUYUUYY

79


<R SEWRGE- ©&xP. |

2s0e 5

zaee 4

1see 4

MERN PPM

sop 4

DAYS

NI SEWRGE—- EXP. 1

“ee 5
ase 4
aoe 4
aso 4
zew 4

MERN PPM

Figure 14. Mean net uptake of Cr and Ni between experimental and
control, sewage - Experiment I.

34


O@T

WERITA INTERNAL
DAY NUMBER 0
CADMIUM = NO -EXPERMENT HEAN= 3.80
CHROMIUM NO EXPERMENT MEAN= 6.23
EOPPER WO EXPEAMENT HEAN] 5S)
NICKLE NO EXPERMENT MEAN= 11.20
LEAD NO EXPERNENT MEAN® 15.90
ratte WO EXPERMENT MEAN= 147.97
DAY NUMBER 1
CADMIUM Ts 600 NOT SIGNIFICANT
CHROMIUM EMEAN=CKEAN
COPPER Ts 200 NOT SIGNIFICANT
NICKLE Ts 000 NOT SIGNIFICANT
LEAD Ts 200 NOT SIGNIFICANT
ZINC Ts 00 NOT SIGNIFICANT
DAY NUMBER 5
CADMIUM EMEAN*CHEAN
CHROMIUM EMEAN=CMEAN
COPPER Ts 200 NOT SIGNIFICANT
NICKLE tT 00 NOT SIGNIFICANT
LEAD Ts 200 NOT SIGNIFICANT
ZINC Ts 200 NOT SIGNIFICANT
DAY NUMBER 25 .
CADMIUM T= 00 NOT SIGNIFICANT
CHROMIUM EMEAN=CMEAN
COPPER Ts 200 NOT SIGNIFICANT
NICKLE tT 200 NOT SIGNIFICANT
LEAD Ts +00 NOT SIGNIFICANT
ZINC Ts 200 NOT SIGNIFICANT
DAY NUMBER 50
CAOMIUK T# 200 NOT SIGNIFICANT
CHROMIUK Te +00 NOT SIGNIFICANT
COPPER Ts 00 NOT SIGNIFICANT
NICKLE tT 290 NOT SIGNIFICANT
LEAD tT +00 NOT SIGNIFICANT
TING Ts 200 NOT SIGNIFICANT
DAY NUMBER 65
CADHIUM Ts +00 NCT SIGNIFICANT
CHROMIUM T= 200 NOT SIGNIFICANT
COPPER Ts 200 NOT SIGNIFICANT
WICKLE Ts 200 NOT SIGNIFICANT
LEAD T= 200 NOT SIGNIFICANT
ZINC Ts 00 NGT SIGNIFICANT
DAY NUMBER 125
CADMIUM = =NU CONTROL = MEAN® 3.80
CHROMIUM NO CONTROL MEAN® 6.30
COPPER Ku CUNTROL =MEAN= 57.70
NICKLE NO CONTROL MEAN® 10,30
LEAD NU CONTROL MEAN® 11.60
zIKC NO CONTROL §=HEAN® 123-20

EXPERIMENT NOs2

DEVIATION=
DEVIATION=
DEVIATIONS
OEVIATION=
DEVIATIGN=
DEVIATIONS

EME AN-CMEAN=

EME AN-CME AN=
EMEAN~CME AN=
EME AN-CHE AN=
EME AN-CME AN=

EME AN-CME AN=
EME AN-CHE AN=
EME AN-CME AN =
EME ACME AN=

EME AN-CME AN =

EME AN-CHE AN =
EME AN-CME AN=
EME AN-CMEAN=
EME AN-CME AN=

EME AN-CME AN =
EME AN-CME AN=
EMEAN-CME AN®
EME AN-CME AN=
EME AN-CME AN=
EME AN-CHME AN =

EME ANW-CME AN=
EME AN-CME AN =
EME AN-CME AN®
EME AN-CME AN=
EME AN-CHE AN=
EME AN-CME AN =

DEVIATIUN=
DEVIATION=
DEVIATION=
DEVIATION=
DEVIATIGN=
DEVIATIGN=

62

22.20

00
00
+00
200
200
+00

veowue

ererererers)

EMEANS
EMEAN®
EMEAN=
EMEAN=
EMEAN=
EMEAN®

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEANS

EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=
EMEANS

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

O EMEAN=

EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN®
EMEANS

120.60

EDEVs
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=

10 EDEV=

EOEV=
EDEV=

200

200
+00

+00
00
+00

CMEAN=
CMEAN=
CMEAN®
CMEAN=
CHEAN=
CHEAN®

CHEAN=
CMEAN=
CMEAN=
CHEAN=
CMEAN=
CHEAN®

CMEAN®
CKEAN=

-CMEAN®

CMEAN®
CHEAN®
CHEAN®

CHEAN=
CHEAN®
CHEANTM
CHEAN®
CMEAN=
CMEAN®

CHEAN=
CHEAN®
CHEAK=
CHEAN=
CHEAN®
CHEAN*

30

1.60

CDEV=

1.70CDEV=

9.60
8.70
9.30
126.10

COEV=
COEV=
CDEV=
CDEV=

1.80COEV=
1.TOCDEV=

61.90
6.50
8.80

176.30

1.70

COEV=
COEV=
CDEV=
COEV=

COEVe

1.70CDEV=

49.00
5.80

6.20
176.70

2.30
7.40
64.80
13.80
10.60
107.40

1.50
10.00
74.90
11.90

6.50

142.60

COEV=
CDEV=
CDEV=
CDEVe

CDEV=
COEV=
CDEVe
CDEV=
CDEV=
COEV=

COEVs
COEVe
COEV=
COEVe
COEV=
COEV=

200
200
200
200


OT

OYSTER SHELL
DAY NUMBER 0
CADMIUM Te 31.00 SIGNIFICANT
CHROMIUM T= +14 NOT SIGNIFICANT
COPPER EMEAN =CHE AN
NICKLE T= 7.00 SIGNIFICANT
LEAD Ts 1.86 NOT SIGNIFICANT
ZINC Ts 11.14 SIGNIFICANT ©
DAY NUMBER 1
CADMIUM T= +63 NOT SIGNIFICANT
CHROMIUM T= 3464 NOT SIGNIFICANT
COPPER EMEAN =CHEAN
NICKLE = T= «95 NOT SIGNIFICANT
LEAD Ts 245 NOT SIGNIFICANT
ZINC Ts 1.57 NCT SIGNIFICANT
DAY NUMBER 5
CADMIUK T= 291 NOT SIGNIFICANT.
CHROMIUM T= 25 NOT SIGNIFICANT
COPPER = T= +00 NOT SIGNIFICANT
NICKLE = Te +05 NOT SIGNIFICANT
LEAD Te 7.82 SIGNIFICANT
ZINC, Ts 6.09 SIGNIFICANT
DAY NUMBER 25
CADMIUM T= +67 NOT SIGNIFICANT
CHROMIUM T= 8.22 NOT SIGNIFICANT
COPPER = T= 1.05 NOT SIGNIFICANT
NICKLE = Tx 7-70 SIGNIFICANT
LEAD Ts 1.28 NOT SIGNIFICANT
ZINC Ts 741 SIGNIFICANT

EXPERIMENT NO=1

EMEAN-CME AN=
EME AN-CAE ANS

EME AN=CME AN =
EME AW-CME AN =
EME AN-CHE AN=

EME AN-CME AN=
EME AN-CME AN

EME AN~CME AN=
EME AN-CMc An =
EME AN-CME AN =

EME AN~CME AN=
CHE AN-CME AN =
ENE AN-CNEAN =
EME AN-ChE Ay =
EME AN-CME AN =
EME AN-CME AN=

EME AN-CME AN=
EME AN-CNE AN=
EME AN-CME AN =
EME AN-CME AN=
EME AN~CME Av =
EME AN-CME AN =

DEG=
DEG=

DEG
DEG=
GEG=

DEGs
DEG=

DEG=
LEG=
DEG=

OEG=
EUs
GEo=
DEG=
DEG

IS DEv=

DEG
DEG=
ODEs
DEG=
DEG=
DEG=

PRNEES RAN NN WINN ORK

NNN

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEANE
EMEAN=
EMEAN=
EMEAN=
EMEANS

EMEANS
EMEAN=
EMEAN®
EMEAN=
EMEAN=
EMEAN=

EMEAN =
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS

5.00

4.40
5.

+
34.90
42.30
13.60

4.35
4.55
4.45
36.95
12.65.
15.40

4.90
4.40
4.45
36.90
46.45
8.65

9.70
170.70
420.55
204.85

41.45
102.45

EDEV= 200 CHEANS
EDEV= 200 CHEAN=
+ EoEVS 80 CHEANS
EDEV= +00 CHEAN=
EDEV= +42 CHEANS
£DEV= 57 CHEANE
EDEV= +07 CMEANS
eDEV= +07 CHEAN=

EDEV= +07. CMEAN=
EDEV= CHEAN=
EDEV= CHEAN
EDEV= ChEAN=
£0EV= CHEAN=
EDEV= CMEANS
EDEV= CHEAN®
eDEV= 1641 CHEANE
EDEV= 5.66 CHEAN=
EDEV= +76 CMEAN=
EDev= +00 CHEAN=
EDEV= 12.73 CHEAN
EDEV= 514400 CHEANS
EDEV= +50 CMEAN=
EDEV= 39.39 CHEAN=
EDEV= wbO-CHEANS

13

3645 COEV=
4.35 COEVE
4 45C0EV=
36.65 CDEV=
40.45 CDEVe
19.90 COEV=

4.45 COEV=
3.80 CDEV=
4eG5CDEV=
37.90 CDEV=
13.00 CDEV=
13.80 COEV=

4.25 COEV=
4.25 CDEVE
445 COEVE
36.95 COEV=
13.15 CDEVe
12.20 CDEV=

3.95 COEVe
42.50 COEV=
40.50 CDEV=
41.50 CDEV=
5.85 CDEV=
77055 COEV=



e€TT

CLAM

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CAOM 1UM
CHKUMIUM
CLPPER
NICKLE
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TING

ADM LUN,
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHRUMIUM
COPPER
NICKLE
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ZINC

CADK TUM
CHROMIUM
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ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
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ZINC

ADM IUK
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

EXPERIMENT NO=2

3.87 OEVIATIUN=
7.17 DEVIAT lune
eeGS DEVEAT IGKE
13.03 DEVIAT IuN=
33.97 DEVIATIUN=
1.90 DEVIAT lune

SHELL
DAY NUMBER 0
NO EXPERMENT MEAN=
NO EXPERMENT MEAN=
NU EXPERHENT HEAN®
NC EXPERMENT MEANS
Nd EXPERMENT MEANS
NG EXPERMENT KEAN=
DAY NUMBER 1
Ts +66 NOT SIGNIFICANT
Ts +53 NOT SIGNIFICANT
Ts 1.00 NOT SIUNIFICART
Te +22 NOT SIGNIFICANT
1s +54 NOT SIGNIFICANT
Ts 1.54 WOT SIGNIFICANT
DAY NUMBER 5
Te 1.00 NOT SIGNIFICANT
Ts NOT SIGNIFICANT
Ts NGT STONIFICANT
Ts NOT SIGNIFICANT
Ts NOT SIGNIFICANT

DAY NUMBER
Te

Ts +40
Te 48
Ts 982
Ts 2.63
DAY NUMBER
Is +72
Ts +06
Ts +14
Ts 2.53
Te 1.28
DAY NUMBER
Tz 245
Ta 54
Ts 227
Ts 225
Ts 1.97
DAY NUMEER
Ts 932
Ts 57
T= 1.02
Te 1.16
Te 49

EMEAN =CME AN
25

0 WOT SIGNIFICANT

NGT SIGNIFICANT
EMEAN =CME AN
NOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT

50
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN=CMEAN
NCT SIGNIFICANT
NUT SIGNIFICANT
WOT SIGNIFICANT
85
NOT SIGNIFICANT
NGT SIGNIFICANT
EMEAN=CMEAN
WOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
125
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN=CHEAN
NOT SIGNIFICANT
NCT SIGNIFICANT
NUT SIGNIFICANT

EME AN-CME AN=
EME AN-CME AN=
EME ACME AN =
EME AW-CNE ON =
EME Aw-CME aN =
EME AN~CME AN=

EME AW-CME AN=
EME AN-CME AN=
ENE AN-CME AN =
ENE AN-CME ANE
EME AN-CME aN =

EME AW-CMEAN =
EHE Ad-Cate Ane

EME AN-CME AN=
EME AN-CME AN=
EME AN~CME AN=

EME AN-CME AN =
EME AW-CME AN =

EME AN-CME AN=
EME AN-CHE AN =
EME AN-CME AN=

EE AN-CME AN=
EME AN-CME AN =

EME AU-CME AN =
EME AN-CME AN =
EME AN-CME AN =

EME AN-CME AN =
EME AN-CME AN =

EME AN-CME AN=
EME AN-CME AN=
GME AN=CHE AN =

+55 NUK=
1.27 NUM
+00 NUKE
1.55 NUN
1.46 NUM=
+87 NUKE

www

+13 DEG=
-227 DEG=
71.48 DEG
~.23 DEG=
72.33 VEG=
71.37 DEG=

-+13 DEG=
=.27 DEG=
1.52 DEv=
-.77 DEG=
-.67 DEG=

+10 DEG=
+17 DEG=

+23 DEvs
1.33 DEG=
3.07 DEG=

-.13 DEG=
+03 DEG=

+07 DEG=
-2.43 DEG=
+93 VEG=

-.03 DEG=
+40 DEG=

+20 DEG=
-+43 DEG=
1.03 DEG=

+03 DEG=
+20 DEG=

+90 DEG=
71.13 VEG=
-.70 DEG=

4
4
4

4
4

4
4
4

4
4

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=

EMEAN=
EMEANS
EMEAN=
EMEANS
EMEAN=
EMEAN®

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEANS

EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=

EMEAN®
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EMEANS
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2.40
6.97
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15.13
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2.67
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(35.13
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23

COEVe
7.23 COEV=
COEV=
COEVe
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COEVe
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35.80 COEVe
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2.07 COEVe
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sTt

MANGROVE LEAF
DAY NUMBER O
CADNIUM = ==NO EXPERMENT MEAN® 37
CHROMIUM NO EXPERMENT MEAN= 1.70
COPPER NO EXPERMENT MEAN® 4645
NICKLE NO EXPERMENT MEANS 5.87
LEAD NO EXPERHENT MEANS 2.85
ZINC NO EXPERMENT MEAN= 15.80
DAY NUMBER 5
CADHIUM T= 036 NOT SIGNIFICANT
CHROMIUM Ts 4082 SIGNIFICANT
COPPER T# 6.53 SIGNIFICANT
NICKLE Ts «58 NOT SIGNIFICANT
LEAD Ts 3.30 SIGNIFICANT
ZINC Ts 1.95 NOT SIGNIFICANT.
DAY NUMBER 50
CADMIUM EMEAN®=CMEAN
CHROMIUM EMEAN SCMEAN
COPPER T= 20.64 SIGNIFICANT
NICKLE Ts Lelk NOT SIGNIFICANT
LEAD Ts 1.51 NOT SIGNIFICANT
ZINC Ts +03 NOT SIGNIFICANT
DAY NUMBER 125
CADMIUM Te 2.12 NOT SIGNIFICANT
CHROMIUM Te 32.91 SIGNIFICANT
COPPER = Tx 2.19 NOT SIGNIFICANT
NICKLE Ts 15.65 SIGNIFICANT
LEAD Ts 1.00 NOT SIGNIFICANT
ZINC Te 3.37) SIGNIFICANT

EXPERIMENT NO#2

DEVIATION=
DEVIATIONS
DEVIATIUN®
DEVIATIGN=
DEVIATIGNS
DEVIATIUN=

EME AN-CME AN
EME AN-CME AN =
EME AWN-CME AN=
EME AN-CME AN =
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EMEAN-CMEAN®

EME AN-CME AN=
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EME AN-CME ANS
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EME AN-CME AN =
EME AN-CME AN®=

+06 NUM&
+00 NUM=
+00 NUM®
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2.37 NUM
1.64 NUM=

~.07 DEG=
78.45 DEGs
7.38 DEG=
—265 DEG=
5.22 VEG=
2.10 DEG=

4.B2 DEG=

+20 VEGS
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1.57 DEG=

010 DEG=
1.90 DEG=
73.47 DEGs
72.33 DEG=
1.25 DEG
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VUVEow

SwUEOW

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Pppees

EMEAN=
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73
1.70
11.63
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EDEVs
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EDEV=

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015 CHEAN®
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1.18 CHEAN®
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28

280 COEV=
10.15 CDEV=
4.45 COEVe
5 COEV=
COEVe
12.20 COEVe

ol SCOEV®
1.70CDEV®
4.45 COEVE
2.47 COEVs
2.85 COEV=
9.50 CDEV=

047
3.60



EPA-600/3-78-035
March 1978

BIOLOGICAL AVAILABILITY OF
POLLUTANTS TO MARINE ORGANISMS

by

John R. Montgomery, Mary Price, John Thurston,
Gina Laite de Castro, Luz Leida Cruz and
Domenica DeCaro Zimmerman

Center for Energy and Environment Research
Mayaguez, Puerto Rico 00708

Contract No. IAG-D-40541

Project Officer

Gerald Pesch
Environmental Research Laboratory
Narragansett, Rhode Island 02882

ENVIRONMENTAL RESEARCH LABORATORY
OFFICE OF RESEARCH AND DEVELOPMENT
U.S. ENVIRONMENTAL PROTECTION AGENCY
NARRAGANSETT, RHODE ISLAND 02882


L6

MANGROVE BooY EXPERIMENT NO=1
DAY NUMBER O
CADMIUM T= +77 MOT SIGNIFICANT EMEAN-CME AN=
CHROMIUM T= 1.00 MOT SIGNIFICANT EMEAN-CMEAN=
COPPER EMEAN @CMEAN
NICKLE Ts 2.94 SIGWIFICANT EME AN-CME AN =
LEAD EMEAN =CME AN
Hac Te 207 NOT SIGNIFICANT EMEAN-CHEAN=
DAY NUMBER 5
CADMIUM T= 1.20 NOT SIGNIFICANT EME AN-CMEAN=
CHROMIUM EMEAN=CHEAN
COPPER EMEAN=CMEAN
NICKLE = T= 200 NOT SIGNIFICANT EMEAN-CME AN=
LEAD EMEAN =CHE AN
TING EMEAN=CMEAN
DAY NUMBER
CADMIUM T= 1.00 NOT SIGNIFICANT EME AN-CHE AN=
CHROMIUM EMEAN SCME AN
COPPER EMEAN=CME AN
NICKLE Ts 000 NLT SIGNIFICANT EME Aiv-CHE An =
LEAD Ts 1.00 NOT SIGNIFICANT EMEAN-CMEAN=
ZINC EMEAN=CMEAN
DA¥ NUMBER 125
CADMIUM NO EXPERMENT MEANS 985 DEVIATIUh=
CHROMIUM NO EXPERMENT KEAN= 2.70 DEVIATIONS
COPPER NO -EXPERMENT MEAN= Ges VEVIATION=
NICKLE NO -EXPERMENT MEAN= 200 DEVIATION=
LEAD NO EXPERMENT MEAN® 3.50 DEVIATIUN=
HNC NO EXPERMENT MEAN® 2.35 DEVIATIOW=

07
+73

37
-.03
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3.00
1.37

222
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200
271
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+50
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1.73
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43 CDEVe
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40 COEVE
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212

200


Ls

Figure 37.

<D THALASS |F ROOTS—- E€XP. 2

MEAN <E-C) -PPM

hy
aa 4
wet 4

DAYS

Mean net uptake of Cd between experimental and control, Thalassia roots - Experiment II.


NI THRALASS !|F LEAVES—- EXP. 2

SB 4
=
& 4
~ se J
a
vv
t q
Ws
- 1B
z 1
ia 4
= iy
- T T
& Bg z
DAYS
Nt URCHIN GUT- EXP 2
ue J
= 4
a
OQ 24
S
cS 1
Y ae |
w
- 1
zoel
fv
wi 4
=
|
e rr é T T T T 1
a Fy

Figure 23. Mean net uptake of Ni between experimental and control,
Thalassia leaves and urchin guts - Experiment II.

43


76

Ch

CADMIUM
CHROMIUM
COPPER:
NICKLE
LEAD
TING

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZIAC

CADM LUM
CHROMIUM
COPPER
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CADMIUM
CHROMIUM
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TINC

CADMIUM
CHROALUK
COPPER
NICKLE
LEAD
TINC

CADW TUN
CHROMIUM
COPPER
WICKLE
LEAD
ZINC

INTERNAL
DAY NUMBER 0
Ts 2466 NOT SIGNIFICANT
EMEAN=CNEAN
T= 12614 | SIGNIFICANT
Ts 1.98 NOT SIGNIFICANT
Ts 1.95 NOT SIGNIFICANT
Ts 1.41 NOT SIGNIFICANT
DAY NUMBER 1
Ts 1.15 NOT SIGNIFICANT
EMEAN=CHEAN
Te 5.25 SIGNIFICANT
Ts +77 NOT SIGNIFICANT
Ts +77 AGT SIGNIFICANT
Ts 6643 SIGNIFICANT
DAY NUMBER 5
Ts 1.00 NOT SIGNIFICANT
EME AN =CNEAN
Te 11.65 SIGNIFICANT
EMEAN=CMEAN
Ts 6610 SIGNIFICANT
Ts 1.18 NOT SIGNIFICANT
DAY NUMBER 25
Ts 1.00 NOT SIGNIFICANT
EMEAN =CME AN
T= 1465 NOT SIGNIFICANT
Ts +10 NOT SIGNIFICANT
Ts 1,00 NOT SIGNIFICANT
T= 11,00 SIGNIF IC AT
DAY NUMBER 59
Ts +82 NGT SIGNIFICANT
Ts 1.08 WOT SIGWIFICANT
Ts +52 NOT SIGWIFICANT
1 2.37 NOT SIGNIFICANT
1s +45 NOT SIGNIFICANT
Ts 1.78 NGT SIGNIFICANT
OAY NUMBER 125
Ts 2635 NOT SIGNIFICANT
Ts +00 NOT SIGNIFICANT
Ts 200 NOT SIGNIFICANT
Ts 257 NOT SIGNIFICANT
Ts +28 NUT SIGNIFICANT
tT +74 NOT SIGNIFICANT

EXPERIMENT NQ=1

EME AN-CME AN=

EME AN-CHEAN®
EME AN-CME AN=
EME AN-CME AN=
EME AN-CME AN=

EME AN-CME AN =

EME AN-CME ANS

EME AN-CME AN =
EME AN-CME aN =
EME AN-CME AN =

EME AN-CME AN =
EME AN-CME AN=

EME AN-CME AN=

EME AN-CHE AN =
EME AN-CME AN=
EME AN-CME AN =
EMEAN-CHE AN=

EMEAN-CME AN=
ENEAN-CME AN =
EME AN-CME AN =
EME AN-CME AN =
EMEAN-CHE AN=
EME AN-CNE AN =

EME AN~CME AN =
EME AN-CME AN=
EME AN-CME AN =
EME AN-CME AN=
EME AW-CME AN =
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1.54

14.57
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—.32

e

Prearearrenrs

iy

PREEDER NWN

assis

EMEAN®
EMEAN=
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EMEAN=
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1

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3.91 COEV=
118.15 CDEV=

2.17 COEV=
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24.05 CVEV=
045 CDEV=
225 COEV=
288.50 COEV=

1.92 COEV=
1.70COEV=
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213.70 COEV=

1.07 COEV=
1.70COEV=
21.30 CDEV=
5.42 COEV=
3.42 CDEV=
93.50 CDEV=

3.37 CDEV=
2.30 CDEV=
6.50 CDEV=
11.67 CDEV=
9.83 COEVs
269.860 CDEV=

1.77 CDEV=
1.70 CDEV=
4.45 COEV=
5.62 CDEV=
5.27 CDEVe
106.72 CDEV=



This was also shown in the field by Camp et al., (1973), The
net uptake of some metals in leaves was closely coupled to the
uptake in the roots of Thalassia. The results for Cd, Ni, and
Cr in Experiment II indicate a translocation of these two metals
from roots to leaves, The translocation of metal was also
indicated for Zn in Experiment II from day 85, where the net
uptake decreases in the roots with an increase in the leaves
and vice versa at day 125. In general, the maximum net uptake
in Thalassia leaves was slightly greater than the uptake in
voots. This is in apparent contradiction of Schroeder's results
(1975) where roots were found to have the greatest uptake. How-
ever, our root sample consisted of roots plus rhizomes and, as
rhizomes showed the highest net uptake in Schroeder's paper
(1975), the contradiction between our results and Schroeder's

is cleared up.

The interpretation of the significant net uptake for mangrove
roots indicates that only Ni and Zn in Experiment I show a near
linear increase over time. In Experiment II, only Cr showed a
linear response over time, whereas, Zn and Cu showed an increase
in net uptake followed by attainment of apparent equilibrium,
then loss of the.metal. Lead shows no net uptake until day 85.
These results appear to be loosely correlated with the fact that
the sediment never showed a net uptake until at day 85 in
Experiment II when Ni, Pb and Zn began to show a slight net up-
take (Appendix B) in the sediment. Since no other part of the
mangroves sampled indicated a significant uptake of metal (Appendix
B), then the only source for the metals in the mangrove roots
must be the sediments. If that is so, then only Ni, Pb and Zn
should show net uptake in mangrove roots and then only after day
85 in Experiment II. This lack of significant uptake of metals
by the sediments could be due to the pooling of whole sediment
samples from a 2.5 cm diam. x 25 cm core. If the gradient of
trace metal concentration in the sediment was very steep, then
the pooling of the core could have diluted the higher surface
metal concentrations with the much lower metal concentrations in
the bottom of the core.

There were similarities in net uptake by holothurian guts
for Experiments I and II. Although there are similar patterns
of uptake in the holothurian gut, the maximum values for net
uptake were always higher in Experiment I for Ni, Cu, Pb and Zn.
Chromium showed a higher maximum net uptake in Experiment II
for holothurian guts. In Experiment II, the sewage sludge showed
a two stage release. The first stage occurred at day 1 followed
by a slow constant release until day 50 when another rapid re-
lease occurred. A slow but steady release would more likely
result in a higher maximum uptake, whereas, a two stage release,
as shown by the sewage sludge in Experiment II would, due to
the estimated 6 hour residence time for the water in the tanks
and 8.5 2 min71 flow rate, produce a lower maximum net uptake.
The data for the net loss of metals from the sewage sludge was

18


Copper, Cr, and Pb show a steady and rapid increase in net
uptake for EXP I and II (Figs. 16, 17, 18, 19, 20, 21). The net
uptake for Cd was not significant in Experiment I (Appendix B),
however in Experiment II (Fig. 26) the net uptake increased after
day 50. The results for nickel and Zn uptake are very similar
in Experiment I and II (Figs. 22, 23, 24, 25, 26). Zinc shows
a steady increase with a decrease, in net uptake, at day 50
followed by a rapid increase in both Experiment I and II.

Sea cucumber, internal organs

In both Experiment I and II, Cr, Cu, Pb and Zn showed very
similar net uptakes over the 125 day period (Figs. 38, 39, 40,
41, 42, 43). Ni showed a net uptake spike at day 5 in Experiment
I, and in Experiment II a spike showing uptake of nickel occurred
at day 5, 50 and a smaller spike at day 125 (Figs. 38, 41).

Mangrove roots

Nickel, in Experiment I, shows an initial decrease in net
uptake before day 50 with a gradual increase to day 125 (Fig. 27).
The opposite occurs in Experiment II where the net uptake increases
rapidly up to day 5 (Appendix B). The net uptake for Cr was not
significant for Experiment I (Appendix B), but indicated a linear
net uptake from day 50 to day 125 for Experiment II (Fig. 30).
Lead uptake in Experiment I and II (Figs. 33 and 34) exhibits
similar trends. However, Pb uptake is much higher in Experi-
ment I (Fig. 33). Zinc, in Experiment I, also demonstrated a
rapid linear uptake curve (Fig. 35) which was very similar to the
uptake in Experiment II (Fig. 36). However, the net uptake was
greater for Zn in Experiment I. The net uptake for Cu was not
significant in Experiment I (Appendix B), but appeared to show
a gradual uptake after day 50 followed by a decline to origin-
al levels (Fig. 32).

Other

No significant net uptake was shown for the oysters, clams,
mangrove body, stem or leaves, snails, holothurian body and muscle
parts or sediment for Experiment I or II. The results shown are
in Appendix B. The significant results for net uptake of metals
are shown in Figs. 6-43 and are indicated by open rectangles.

Dark rectangles indicate non-significant results.

14


96

EXPERIMENT NO=1

HOLOTHUR MUSCLE
DAY NUMBER 1
CADMIUM Te +48 NOT SIGNIFICANT
CHROMIUM Te 1.97 NOT SIGNIFICANT
COPPER Ts 0 NUT SIGNIFICANT
NICKLE Te 2.46 SIGNIFICANT
LEAD Ts «15 NOT SIGNIFICANT
ZINC Ts 1.41 NOT SIGNIFICANT
DAY NUMBER 5
CADMIUM Te 287 NOT SIGNIFICANT
CHROMIUM T= 1.15 wOT SIGNIFICANT
COPPER Te 200 NOT SIGNIFICANT
NICKLE Ts 1.41 NOT SIGNIFICANT
LEAD Ts 47 NUT SIGNIFICANT
ZINC Te 2.09 NOT SIGNIFICANT
DAY NUMBER 25
CADMIUM T= 2.00 NOT SIGNIFICANT
CHROMIUM EMEAN=CMEAN
COPPER EMEAN=CMEAN
NICKLE EMEAN =CME AN
LEAD Ts +48 NOT SIGNIFICANT
ZINC Ts 1.48 NOT SIGNIFICANT
DAY NUMBER 50
CADMIUM NO EXPERMENT MEAN= 1.65
CHROMIUM NO EXPERMENT MEAN= 6.35
COPPER © -NO EXPERMENT NEAN= 4645
NICKLE NO EXPERMENT MEAN? 12.22
LEAD NO EXPERMENT MEANS 11.15
ZINC WO EXPERMENT MEAN= 41.87
DAY NUMBER 125
CADMIUM T= 1.24 NOT SIGNIFICANT
CHROMIUM T= 081 NOT SIGNIFICANT
COPPER Ts 200 NOT SIGNIFICANT
NICKLE Ts 1.85 NOT SIGNIFICANT
LEAD Ts 2611 NOT SIGNIFICANT
ZINC Ts +48 NOT SIGNIFICANT

EMEAN-CHEAN ©
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+00 CHEANS
+00 CHEAN=
+09 CHEAN=
1.84 CHEAN=
+28 CHEAN=
214 CHEAN=
w42 CHEANS
CHEAN=
CMEAN=
CHEAN®
CHEAN?

5

1.44
2.67
4.45
10.30
8.30
55642

17
1.70
4.45
8.55
9.70

45.55

292

COEV=
CDEV=
COEV=
CDEVe
CDEVe
COEV=

COEVe
COEV=
COEVe
COEV=
CDEVE
COEVe

CDEVe

1.70CDEV=
445 CDEV=
3.65CDEV=

13.55
64.35

16.35
11.67
45.95

CDEVe
CDEV=

CDEV=
CDEV=
COEV=
CDEVe
COEV=
CDEVe

3.18

1.27
2.41

4eb1
5655
10.38


CDRP, EXPERIMENTAL EXP 1

.
4.S 4
3.84
iw 4
“XN
Er 2.s4
L
5 4
2 1.s 4
= 4
Sot
= & g Fy
CDAYS

DRP- EXPERIMENTAL EXP.2

es]
y s.s4d
iE 4“.S 4
wh ae]
S 2c]
1.5 4
= 4
i
a xo o ivi
a s g
CDRYS
Figure 5. Dissolved reactive phosphate concentration

Experiment I and II, experimental. Data points
are mean values for 5 day period, excluding day 1.

25


90T

SEDIMENT
CAY NUMBER
CADH 1UH
CHROMIUM Te 141
COPPER Ts 2
NICKLE = T= 4.09
LEAD Ts 37
ZINC Ts 2.09
DAY NUMBER
CADM LUM
CHROMIUM
COPPER = T= 1.41
NICKLE Te 1,06
LEAD
ZINC +26
DAY NUMBER
CADMIUM
CHROMIUM T=
COPPER = T=
NICKLE T=
LEAD Ts
ZINC 1s
DAY NUMBER
CADMIUM T= 1.00
CHROMIUM T= 47
COPPER «T= +05
WICKLE = T= 3.71
LEAD Ts +43
ZINC Ts 59
DAY NUMBER
CADMIUK 12 +97
CHRQMIUM T= 1.79
COPPER
NICKLE T= +74
LEAD Te +00
ZINC Ts 1.41
DAY NUMBER
CADMIUM T= 2.61
CHROMIUM T= 81
COPPER = T= 1.56
WICKLE = Ts +04
LEAD T= +83
ZINC Ts 1.25

EXPERIMENT NO=1

0

EMCAN=CREAN
NOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
NGT SIGNIFICANT
WOT STGNIFICANT

NCT SIGNIFICANT

NOT SIGNIFICANT

NOT SIGNIFICANT

WOT SIGNIFICANT
5

EMEAN=CMEAN
NOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
NGT SIGNIFICANT

NOT SIGNIFICANT
NOT SIGNIFICANT
NGT STONIFICAKT
NOT SIGNIFICANT
WOT SIGNIFICANT
WOT SIGNIFICANT
50

NOT SIGNIFICANT
WOT SIGNIFICANT

EMEAN=CHEAN

NOT SIGNIFICANT
WOT SIGNIFICANT
NGT SIGNIFICANT
125

NOT SIGNIFICANT
NOT SIGNIFICANT
wCT SIGNIFICANT
NGT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT

EME AN-CMEAN=
EME AN~CME AN
EME AN-CME AN =
EME AN~CME ANS
EME AN-CME ANS

EME AN-CHE AN =
EME AN-CME AN=
EME AN-CME AN=
EME A-CMEAN =

EME AN-CME AN
EME Aw-CME AN=
EME AN-CME AN =

EME AN-CME AN =
ME AN-CME AN=
EME Aiv-CME AN =
EME AN-CME AN =
EME Aw-CME AN =
EME AN-CME AN=

EME AW-CME bw =
EME AwW-CKE At

EMEAN-CME AN=
EMEAN-CMEAN®
EAE AN-CME AN=

EME AN-CME ar

EME AN-CME AN=
EME AN-CME Al
EME AN-CME Al

1.00
045
4.00

3.15

-.95
71.00
205
225

~222
-.55
227
-3.90
-3.05
-1.50

245
1.70

73.50
00
— 185

72645
71.20
71.05
+05
1.40
73.50

DEG=
DEvs
OEG=
DEG=
DEG

EMEAN=
EMEAN=
EMEAN®
EMEAN=
EMEAN=
EMEAN=

RRA

RNA

EMEAN=
EMEAN=

EMEAN=
EMEA
EMEAN=
EMEAN=
EMEAN=
EMEANS

RNR RNR NANA

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

NN

EMEAN=
EMEAN=
EMEAN=
EMEAN=
2 EMEAN=
2 ENEAN=

21S
6.50
41.45
4.00
14.45
23.30

215
7.50
10.45
8.00

ry
20.95

215
6.00
045

5-50
12.95
16.15

1

215
6.95
10.45
6.95
13.90
19.65

365
7.20
4.45
9.40

10.00
18.40

1.95
10.70
10.45
16.90
11.95
16.25

200
71
78
00

3.61

éel2

2.12

2082
264
1.41
5.59
1.34

1.34
1.61
76
2.12
2.90
1.48

200
1.48
+64
07
1.41
1.34

07
42
82
228
+00
57

1.20
1.41

264
1.41
2.05
1.63

CMEAN=
CMEAN=
CHEAN®
CMEAN=
CHEAN=
CAEAN=

CMEAN=
CMEAN=
CMEAN=
CMEAN=
CHEANS
CMEAN=

CMHEANS
CMEAN=
CMEANS
CMEAN=
CMEAN]
CMEAN=

(MEANS
CMEAN=
CHEANS
CMEAN=
CMEAN=
CHEAN=

CMEAN=
MEAN=
CMEANS
CMEAN=
CMEAN=
CMEAN=

MEANS
CMEAN=
CMEAN=
CHEAN=
CMEAN=
CHEAN=

1é
o L SCDEV= 200

5.50 CDOEV= o7l
11.00 CDEV= 200
8.00 COEV= 1.41
13.50 COEV= 71
20.15 COEVE o2h

sl SCDEV= +00
7.50C0EV=
11.40 CDEV=
9.00 COEV=
14.90 COEV=
20.70 CDEV=

+ 1SCOEV=
6.50 CDEV=
11.45 CDEV=
9.05 CDEV=
17.90 CDEV=
20.90 COEV=

37 CDEV= +32
7.50 COEV= 71
10.17 COEV= 8.10

10.85 CDEV= 1.48
16.95 CDEV= 9.63
21.15 CDEV= 3.32

2.10 CDEV=
5.50 CDEV=
4.45CDEV=
12.90 COEV=
10.00 CDEV=
19.25 CDEV=

4.40 CDEV=
11.90 CDEV=
11.50 CDEV=
16.95 CDEV=
10.55 CDEV=
21.75 CDEV=



LET

THALASSL

CADMIUM
CHROMIUK
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM 1UK
CHRUMIUM
COPPER
NICKLE
LEAD
ZINC

ADH TUM
CHROMIUM
COPPER
NICKLE
LEAD
LINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM IU
CHROMIUM
COPPER
NICKLE
LEAD
ZIKC

LEAF EXPER
DAY NUMBER O
NO EXPERMENT MEAN® 1.1

NO EXPERMENT MEAN=

NO EXPERMENT HEAN= 464
NO EXPERMENT MEAN= 2964
NO EXPERHENT MEAN= 10.0
NO EXPERMENT KEAN= 27.6

1
NOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT

5
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
25
SIGNIFICANT
SIGNIFICANT
SIGWIFICANT
SIGNIFICANT
SIGHIF ICANT
SIGNIFICANT
50
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NGT SIGNIFICANT
85

SIGWIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT

SIGNIFICANT
SIGiWIF ICANT
SIGNIFICANT
SIGNIFICANT
EMEAN=CHEAN

DAY NUMBER
Te +89
1s 2.28
Ts 1.49
1s 1672
Te 10.42
Ts +24
DAY NUMBER
1s 80
1s 4.40
Te 8.59
Ts 9465
Ts 4.50
Ts 335
DAY NUMBER
Ts 4.16
T= 23.57
Ts 6.54
Ts 17.56
1 10.73
Ts 9.63
DAY NUMBER
T= 8.19
Ts 4.93
Ts 2.59
Ts 8.67
Ts 5.27
1 1.88
DAY NUMBER
Ts 0.84
Ts 17.41
Ts 11.36
Ts 53412
Ts 59
1s 3.13
DAY NUMBER 125
Txsesseeone
Ts 9.37
Tx 16.62
Ts 13,87
Ts 3.12

NOT SIGNIFICANT

IMENT NO#2
O DEVIATION=

16.13 DEVIATION=

5 DEVIATION=
© DEVIATION=
7 DEVIATION=
3 DEVIATION=

EME AN-CMEAN=
EME AN~CME AN®
EME AN~CNE AN
EME AN~CME AN=
EME AN~CMEAN®
EMEAN~CME AN®

EME AiN~CME AN =
EME AN~CHE AN =
EME AN-CME AN=
EME AN~CME AN=
CME AN-CME AN =
EME AN-CME AN=

EME AN-CME AN =
ENE AN-CME AN=
EME AN-CME AN =
EMEAN-CME AN=
EME AN-CMEAN=
EME AN-CME AN=

EME AN-CME ANS
EME AN-CHE AN=
EME AN-CME AN =
EMEAN-CME AN=
EME AN-CHE Al
EME AN-CME AN =

EME Aw—CME AN =
EME AN~CME AN =
EME AN-CME AN=
EME AN-CNE AN=
EME AN-CMEAN=
EME AN-CMEAN=

EME AN-CME AN=
EME AN-CHE AN=
EME AN-CME AN®
EMEAN-CME AN®

EME AN-CME AN

210
257
200
1.30
264
292

225
2.32
272
3.50
5.60
-.37

223
14.03
3.73
14.40
4.80
80

57
~5.37
72042

712.23
7260
~8.67

3.10
16.40
5.70
717.13
3.67
6.70

983
719.87
5.77
42.90
213
6.87

2.30
21.97
14.53

7.57

3.30

NUM®
NUM=
NUM
NUM=
NUM=
NUM=

DEG=
DEG=
OEG=
DEG=
DEG=
DEG=

DEG=
DEG=
DEG=
DEG=
O&G=
DEG=

OEG=
DEG=
DEG=
DEG=
DEG=
DEG=

DEGs
DEG=
DEG=
DEG=
DEG=
DEG=

DEG=
OEG=
DEG:
DEG=
DEG=
DEG=

DEG=
DEG=
DEG=
OEG=

DEG=

wwewww

WEOWUb SEESEE HHOWUOL

Presper peeees

Ny ONAN

EMEANS
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEANS
EMEANS
EMEANS
EMEAN=

EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN®

1.70
15.47
10.87
24.20
12.20
25.83

2630
53.30
18.97
56.63
14.27
37.33

4.20
32.00
20.15
29.50

7.85
23.20

5.80
39.53
30.60
27.10
13.77
47.07

3.47
124.87
29.17
13.67
6-60
31.80

3.50
73.20
29.60
36.60

225
15.00

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEVe

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEVe
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

235
1.14

CMEANE
CMEANS
CMEAN=
CMEAN=
CHEAN=
CMEAN®

CMEAN=
CMEAN=
CMEAN=
CHEAN=
CMEAN=
CMEAN=

CMEANS
CMEAN®
CMEANS
CMEAN=
CMEAN=
CMEANS

CMEAN=
CHEANS
CHEAN=
CMEAN=
CHEAN
CMEAN=

CHEANS
CMEAN=
CMEAN=
CHEANS
CMEAN®
CMEAN®

CMEAN=
CMEAN®
CHEANE
CHEAN=
CMEAN=
CHEAN®

37

o25CDEV=
11.70 CDEVs

22k
1.06
07
71
71
2.55

o2l



6p

Figure 29.

CR THALASS!|A ROOTS—- ©&xP. f

3a 4
=
a 4
a
S
22 4
a
vu
Y 4
im)
uv 124
z
r 4
ui
= a 4
| a ee ee ee
s é g a
DAYS

Mean net uptake of Cr between experimental and control, Thalassia roots - Experiment I.


€OL

ADM IUK
CHROMIUM
COPPER
NICKLE
LEAD
ZIKC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUh
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
KICKLE
LEAD
ZINC

OYSTER IATERN AL
DAY NUMBER 0
Ts Ll NOT
Ts 37 NOT
Ts 1.30 WOT
T= 245 NOT
Te 219 NOT
Ts 274 WOT
DAY NUMBER 1
Ts oO NOT
Ts NOT
Ts NOT
Te NOT
Ts not
Ts 200 NET
DAY NUMBER = 5
Te +00 NOT
Ts +00 NOT
Ts 00 NOT
Ts +00 NCT
Ts 200 NCT
Te 90 NOT
DAY NUMBER 25
Ts . inGT
Te +00 WOT
Ts 209 NGT
tT 200 wht
Ts 00 NCCT
Ts: 0 NOT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
DTGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGHIFICANT
SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
STONTFICANT
SIGNIFICANT
SIGNIFICARKT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
STONTFICAKT
SIGNIFICANT

EXPERIMENT ND=L

cMEAN-CME AN =
EME AN-CME AN =
EME AN-CiKE AN =
EME AN-CME AW =
EME AN-CHE AN
ENE Aa-CME AN =

EME AN-CME Ail =
EME Au=CME AN =
EME Aw-CAE ANS
EME AN-CME ZI
EME AN-CME aiv=
EME AN-CME aN =

EME An~CME An =
EME AN-CME aN =
EAE Aw-CME AN =
EME An-CME Ai
EME AN-CME ah
EME AN-CME ai

EME ANCE AN =
EWE AN-CME AN =
EME AN-CNE ANS
EME ACME All =
EWE AN-CME AN =
EME AN=CME ah =

1S
-1.55
204.55
2.15
1.15
416.05

~.10
-2.50
99.30
2.00
1.20
140.20

1.20
2.20
202.70
6.10
30.50
7622.40

3.00
+10
257.60
23.40
27.80
8.40

DEG=
DEG=
DEL
DEG=
DEG=
DEG

OEG=
OEG=
OEG=
LEG
beG=
LELs

DEG=
DEG=
OEG=
vos
DEG
vEGs

DEG=
GEGs
VEG=
DEGS
DEG=
DLG=

ecco oe NANA

©cec0e CoCece

EMEANS
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEANS
EMEAN =
EMEAN
EMEAN
EMEAN
EMEAN

EMEAN=
EMEANS
EMEAN=
EMEAK
EMEAN
EMEAN=

ENEANS
ENEAN=
EMEAN=
cHEANS
EMEAN=
EMEANS

2.65
3.75
573.05
11.80
10.75
1315.40

2.50
1.70
220.90
10.90
8.80
1057.00

4.00
3.90
87.90
13.20
36.20
527.60

6.99
4.00
363.90
30290
33.50
611.70

EOEV=
EDEV=
EDEV=
cDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDtV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EbDEV=
EDEV=
EDEV=
EDEV=
EDEV=

207

2.90
57.77

5

392.87

200
200
00
00
+00
+00

200
00
00
+00
00
200

200
200
00
+00
200
00

CMEANS
CMEANS
CHEAN®
CHEAN=
CAEAN=
CAHEANS

CHEAN=
CMEAN=
CMEAN=
CMEANS
CMEAN=
CHEAN=

CHEAN=
CMEAN=
CMEAN=
ChEAN=
CMEANS
CHEAN=

CMEAN=
CHEANS
CMEAN=
ChEAN=
CMEAN=
CHEAN=

12

2.50
5.30
368.50
9.65
9.60
899.35

2.60
4.20
121.60
8.90
10.00
916.80

2.80
1.70
290.60
7.10
5.70
1150.00

3.90
3.90
106.30
7.50
5.70
620.10

CDEVe
CDEV=
CoEV=
COEV=
COEV=
COEV=

COEVe
CDEVe
CDEV=
CDEVs
COéV=
COEV=

COEVE
COEV=
COEVs
CDEV=
COEV=
CvEV=

COEV=
COEV=
CDEV=
cDEVe
COEVe
Cbeve

1.98
5.09
214.25
6.72

+00
00
00
00
+09
+00

+00
200
+09
+00
200
200

209
200
+00
+00
209
200


Valiela, I., M.D. Banus, and J.M. Teal, 1974. Response of salt
marsh bivalves to enrichment with metal-containing sewage
sludge and retention of lead, zinc, and cadmium by marsh
sediments, Environ. Pollut, 7: 149-157.

Vallentyne, J.R. 1957. The molecular nature of organic matter
in lakes and oceans with lesser reference to sewage and
terrestrial soils. J. Fish. Res. Bd. Canada 14(1): 33-82.

Vinogradov, A.P. 1953, The elementary chemical composition of
marine organisms. Sears Foundation New Haven, Conn.

Zimmermann, C., M. Price, J. Montgomery. 1977. Operation, methods
and quality control cf Technicon AutoAnalyzer II systems for
nutrient determinations in seawater. Harbor Branch Found.
Tech. Rep. 11. 53 p.

67


SECTION 5
CONCLUSIONS AND RECOMMENDATIONS

Net uptake of trace metals by members of a Thalassia
testudinum/Rhizophora mangle community was demonstrated in two
125 day replicated experiments over the period from March 1975
to April 1976. The net uptake was especially significant in:
Thalassia testudinum leaves and roots/rhizomes, internal organs
of the urchin Lytechinus variegatus, "fouling organisms", internal
organs of Holothurea sp., and for some metals in Rhizophora mangle
roots. In general, the net uptake pathways followed the trophic
levels in the food webs. This pathway was especially noticeable
from the water to the fouling organisms and from the water to
the Thalassia testudinum leaves and thence to the urchin herbivore
Lytechinus variegatus. The net uptake in the Holothurea sp.
was closely related to the net uptake in the fouling organisms
and the trace metal loss rate of the sludge. The food web in
this artificial system was dominated by the fouling organisms as
would be expected. The uptake of trace metals, leached from the
sludge, was always greatest in this complex group.

The results of long term exposure, in a simulated tropical
ecosystem, to toxic trace metals more closely approximate the
situation in nature than do laboratory experiments. In this
respect we are in agreement with the statement by Phelps et al.,
(1975) "Chronic exposures to elevated metal levels through the
food web should be used as a more realistic test for possible
deleterious effects from such metals as Cr". No artificial
system can ever reproduce a natural ecosystem. However, it is
more important to closely approximate the natural system with a
system which can duplicate itself (Menzel, D.W. 1977). Our
results show that chronic exposure to toxic metals can lead to
uptake and concentration of these metals in marine organisms.
We were able to duplicate our results using this complex simu-
lation of a marine ecosystem.

More detailed work on the net uptake of toxic metals, using
this complex simulated ecosystem, and the roles of the "fouling
organisms" and plankton in this system is needed. This must
involve integrated sampling of the organisms as well as the water.
The advent of more precise methods of direct trace metal determi-
nation in sea water using. anodic stripping voltammetry and non-
flame atomic absorption spectrophotometry, in lieu of flame AAS,
should allow a reasonable sample size for both organisms and

water.

20


wvvvVVOOOOOOMOHDCOCOCFMOACOFFOOMOOFCeOGAoOeAaGcoCmMoOoOSTDAGGGe0CaDG

VOODOO STUHUVUEY ND YNHYY HYUN YU.WUH DDD UY WUD WWD WU ee et ee it te tet

NN NNDNNNNN NNN NANNY N NUNN NNNNNNURNNNNNNNNNNNNNNNONNNN RAND
PR ROC OP RP HOD ORR HOD ORE HOO ORE H OOO HH HOO OPH H OOO REE MOOR OME

SORTED DATA

50
50
65
65
125
125
125

VUUUU Ue ee eee OOO

3.1
2.0

°
>

errr ar wr weer er ar or ee er ae ee ey

eee

WAG OCSBOLWHRPWWODINNNDROOODNYNRDODONNDOHEE ROE NUROHNE MOUWOS @

eR RR EU N NN NRE ER RE ER ENR EEUU NNMUWWWWUNWYWWNWHWWWWNW WD

eee eeo eee eset

85

oe eer ee eee

OCRANOP SRS PLOY NABABROMONFON ORO

~
SVN SVP OWVMIMAOVCVC OO DNMPONNORATDOS

17.3
15.6
25.0
11.0
22.4
28.4
16.9
17.8
16.6
18.4
17.0
16.8
17.8
20.8
21.5
21.5
1@.0
15.9
19.0
18.2
18.3
21.2
14.4
14.3
16.4
20.1
16.0
18.1
12.8
13.9
14.1
14.0
12.7
13.4
14.6
14.3
15.2
13.0
14.7
15.8
25.0
23.6
23.8
20.8
20.8
21.7
338.0
318.8
354.5
262.5
288.6
296.7

37.7

32.1

me


SECTION I
INTRODUCTION

The deposition of sewage sludge in the marine environment
may adversely affect the resident organisms in three ways:
(1) Siltation with resultant death of sessile organisms. (2)
Production of an anoxic environment due to the oxidation of
the large organic load in the sludge. (3) By the release of
toxic materials into water in forms that may be incorporated
into the organisms. These materials usually include metals
(zinc, cadmium, chromium, nickel, lead and copper), pesticides
or organohalogens and hydrocarbons of petroleum origin. The
mass dumping of treated sewage sludge into oceans and shallow
inshore areas is occurring in a number of areas. The New York
Bight, is an example of an area receiving large quantities of
solid waste for almost a century (Carmody et al., 1973).
Carmody, Pearce, and Yasso (1973) determined that there was a
definite increase in the sediment of Cr, Cu, Pb, Ni and Zn due
to the dumping of sewage sludge. The concentration of Cr, Cu,
Cd, Pb, Ni, and Zn in sewage sludge is markedly elevated over
ambient marine water concentrations (Salotto, B.V. and Farrell,
J.B., 1971; Jacobs, S.A., 1973). Therefore, the deposits of
this sludge in the marine environment could result in the up-
take and concentration within the food web. The uptake of
metals by marine organisms and concentration within the food
web has been demonstrated numerous times (Jacobs, S.A., 19733
Kerfoot and Jacobs, 1973a, 1973b; Kerfoot, W.B., 1973; Shuster
and Pringle, 1969; Valiela et al., 1974).

The most extensive compilation of metal concentrations in
the tissues of marine organisms is found in Vinogradov (1953)
and Goldberg (1965). However, the results were usually based
on very small sample sizes so it is very difficult to assess
the background levels of metals in marine organisms. Eisler
(1973) compiled an annotated bibliography on biological effects
of metals on aquatic organisms. Montgomery et al. (1976) found
that in order to obtain a valid sample size for trace metal
determination in the thread fin herring (Opisthonema oglinum
Le Sueur)) a pooled sample size of from 50 to 124 fish was
necessary in order to detect a 15 to 25 percent difference in
two populations. Other researchers have shown the effect of
various metals on the marine food web or individual organisms
both in laboratory models and field experiments (Phelps, D.K.
et al., 1975; Nair, K.V. et al., 1973; Eisler, R. et al., 1972;


APPENDIX A

Sorted Raw Data Experiment I and II

There are eleven (11) columns shown which represent the
following:

Column
1 - Animal type

Clam

Holothurian
Mangrove
Thalassia
Fouling Organisms
Urchin

Sediment

NAVHEDO
wnnnm nn

2 - Tissue type

M = Muscle

I = Internal contents (guts)
S = Shell

B = Body/Bottom

L = Leaves

R = Roots and rhizomes

G = Upper Meristem

D = Solids

3 - Experiment number - I(1) or II(2)

4 - Sample type - whether from experimental tank (0) or
control tank (1)

5 - Represents sample time

0 = presample
1 = 1 day after sewage added
5 = 5 days " " "
25225" " " "
50 = 50 " " " w
85 = 85 " " " "
12 5 = 12 5 " m w t

68


SOKNDH MDH DAOOAMNADAOCH TE MOSOWH ANDAMAN ON SHOOMONRO TONE EVIIAOOS
ca Daa ea rk a er eee)
NOCHE NOONE ANODE BSMARR ALDOR BSANDARSENMANTOR ADAM N NOMS
aaa nea NANAANDFNMMANE HN OADOFO aa a aes

adda dade

SPOKOGOOCDDDD DADA OAARADMALRNENEOAKVOKACONB®OEHROWOCMROVBMLOYT
coe een eee ee epee reo reer er eee eer eee ereeeeeresranseepeereseeeeeneerd
OONF SFT NV®LPONAOCHOVDOOMNOODTTOOHNRT DONO TNAODAANADNANAL AMOS

ae FAUNANMM A ORE ETRE RR ON OUE RARER OTT NS

SLSSVSPASSHSCOHARSLA|HI LT HORAK IM AAA QOS AATBAAROTSATASHANO #0
seek eee eee e Pe Rr re RE re a et
«© CORA BERD SOR NAF HOR OMAREAATATAGAT THASHSANMAANAM
AA DAANNANKRABAH HAMOMDAKHFRAFHR KPASHRAMM MM my
an BAD Oo POESAARD LHAOAD
totad ta rerrrre ovprvrard
RE LOPAONE GOH LOBMNBONAANAVTMNOVON EH ANHCARLOOCSZOORROTOS CON
rere are cee eee eee eer eres ier ere ear era a ar re) : cree)
NOON TMMOMNTTSFTFOOTITMHONADOOMRMDARAGONHAARHMHSFOGDANTHAE HOODOO D
TFNONMAN SHANE oo ae ao
ao Oo nO Oa
ta i) vt 14
7.)
ANAHRHOKRHRRSCOCODORN HON AHAHRSLORONAQNHAMHAAATANAARAAHA TH AM AM
econ ee EC BS IE SS Sg
a aan aaa BEAFASMARAAPAFARAGGHARAARRTIAAARAHAAAN GS
BD ABR ARH BAAD ALAN DARRAAARH BAKRAKH
SB BAR AAD DA AD a AO PTRHRBAKRAR BAKRAAK
to opeet tee rede tot thrraree Preven
=a
&
FVODVAVN AOD DOF ONNVITARTAGEAVOTALHESARA SESE NARAAINN OAS
Qeeerscveces 2 PP i a Pa a ae Bee ir ee a ee Ee
BO AAA HAAN ARM ASATMMASAW GE FASTSTOEMARAGKTTEMNG
o z Seon naan AHFFFACHARAR ARBOR
w SOR oO FFARSAASFHR ROKOS
- ote tt rerrtrnres i ir
a
a
AONNKHNNNNNKHODDCONNNNNODOANNNHNOOCKHODSCCOCOCBMAAANNNHNNMNMNNOODSDOD
WANNA WMNnA NNN NN NNN NNN AHA
anaes Atte I

AQDOKR HOOD HA AQOABHOOHHAHOHHOTNOBOHOOODOHANOONAOOR NGO MN MOOM tte

EN EE EL DM Et LL dE NL A tt ed tt od tt tt td
St EEE EE NNNNNNNNYNNNNNNNHNNNNNNH
ercrarecexreexceerrrexrzz zx zezzzzzzzz2z2zzz2z2ezzzzzzzz2zzzz2zzz

74


14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

Mean net uptake of Cr and Ni between experimental
and control, sewage - Experiment I..............5-

Mean net uptake of Cr between experimental and
control, fouling organisms and sewage-

Experiment Il... .. eee eee eee eee eee eee eee eee

Mean net uptake of Cu between experimental and
control, Thalassia leaves and urchin guts-
Experiment Tisvii ec cc ccc cece reece eee e ccc eeeee sees

Mean net uptake of Cu between experimental and
control, Thalassia leaves and urchin guts-
Experiment IT... ............- see ee eee eee eee

Mean net uptake of Cr between experimental and
control, Thalassia leaves and urchin guts-
Experiment Lovee cece cee eee eee e teen eee

Mean net uptake of Cr between experimental and
control, Thalassia leaves and urchin guts-
Experiment TDs i.e ec cc cece ec eee e eee e eee

Mean net uptake of Pb between experimental and
control, Thalassia leaves and urchin guts-
Experiment Tov wee ccc ec cece eee eect eee tees

Mean net uptake of Pb between experimental and
control, Thalassia leaves and urchin guts-
Experiment IT........ wwe eccee wc meen eeeee see e cece

Mean net uptake of Ni between experimental and
control, Thalassia leaves and urchin guts-
Experiment I...........- sec eee eee eee eee e eee eee

Mean net uptake of Ni between experimental and
control, Thalassia leaves and urchin guts-
Experiment ITs... cece ec ce cee ee eee etree eee

Mean net uptake of Zn between experimental and
control, Thalassia leaves and urchin guts-

Experiment [ev cece cece cee cere reece eee tee eeeeneee

Mean net uptake of Zn between experimental and
control, Thalassia leaves and urchin guts-

Experiment IT... eee ccc cee eee eee eee eee eee

Mean net uptake of Cd between experimental and
control, Thalassia leaves and urchin guts-
Experiment IT.......... eee ee ee eee eee ee neces

Mean net uptake of Ni between experimental and
control, Thalassia roots and mangrove roots-
Experiment Tove ccc cece ee cece tee eee eee e eee eeee

Mean net uptake of Ni between experimental and
control, Thalassia roots and mangrove roots-
Experiment II.. cee reese err c ces esencse scone

Mean net uptake of Cr between experimental and
control, Thalassia roots - Experiment I...........

vii

+34

335

336

38

39

40

A2

a7

As

Ag


Zot

NERITA SHELL EXPERIMENT NO=1

DAY NUMBER oO

CADMIUM Ts 1.15 NOT SIGNIFICANT EMEAN-CME ANS

CHROMIUM Te 1.25 NOT SIGNIFICANT EMEAN-CMEAN=

GEPRER te 200 NOT SIGNIFICANT EHEAN-CME ANS

NICKLE Ts 1.15 NOT SIGNIFICANT EMEAN-CME GSI

LEAD Ts o91 NOT SIGNIFICANT EME ANCE ON

wine Ts 072 NGT SIGNIFICANT EME AN-CAE AN=
DAY NUMBER 1

ADM TUN EMEAN=CHE AN

CHROMIUM EMEAN=CMEAN

COPPER EMEAN=CNE AN

KICKLE EMEAWSCHE AN

AAD Te S21) SIGWIFICANT EME AN-CNe AN=

ZINC Te 4.67 SIGNIFICANT EMEAN-CME AN =
DAY NUMBER 5

CADMIUM T= 1-00 NOT SIGNIFICANT EMEAN-~CME AN=

CHROMIUM T= 1.00 NOT SIGNIFICANT EME AN-CMEAN=

COPPER EMEAN=CMEAN

NICKLE ENEAN =CHE AW

LEAD Ts 241 NOT SIGNIFICANT EME AN~CME AN=

LINC Ts 1.02 NOT SIGNIFICANT EMEAN~CMEAN=
DAY NUMBER 25

CADMIUM EMEAN=CHE AN

CHROMIUM ENEAN=CEEAN

CUPPER EXEAW =CKEAN

NICKLE EMEAN =CHEAN

LEAD Ts 1.33 NOT SIGNIFICANT EMEAN~CME AN=

ZINC Ts 1434 NGT SIGNIFICANT EME Aiv~CNEAN=
DAY NUMBER 50

CADMIUM Ts +42 NOT SIGNIFICANT EMEAN~CME AN

CHROMIUM Ts «26 NOT SIGNIFICAKT EME AW~CHE AN=

COPPER tT 200 NOT SIGNIFICANT EMEAN~CMEAN=

NICKLE Te +45 NOT SIGNIFICANT EMEAN~CMEANS

4EAD Ts +23 NOT SIGNIFICANT EME AWw-CME ANS

ZINC Te «52 NOT SIGNIFICANT EMEAN-CME AN]
DAY NUFBER 125

CADAIUN NO EXPERMENT HEAN= 4.35 DEVIATIONS

CHROMIUM NO EXPERMENT KEAN= 3.45 OEVIATIOK=

COPPER NO EXPERMENT MEAN= 4-45 DEVIATION®

NICKLE. NO EXPERMENT MEAN 33-90 DEVIATIONS

LEAD NG EXPERMENT MEAN= 42-45 OEVIATION=

ZINC NU EXPERMENT MEAN? 7.00 DEVIATIDN=

2.72 DEG=
2247 DEG=
-.00 DEG=
17.17 DEG=
-12.38 VEG=
7.90 VEG=

14.35 vEG=
~2.85 Deo

~1.67 DEG=
1.50 DEG=

6.10 DEG=
-1.05 DEG=

3.25 LEG=
-1.90 DEU=

-+23 DEG=
7255 vEv=
+00 DEG=
+20 DEG=
=.77 DEG
7.37 DELS

207 WUM=
207 NUM=
+00 NUM=
200 NUN
1.91 NUM=
+28 NUMs

RNA

EMEANS
EMEANS

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

45
72.70
10.60

4.25
3.60
4.45
33.15
43.20
7.10

EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

3.15
2.86
00
19.85
18.17
1.66

1.66
1.66
1.66
1.66
1,63
271

00
2.12

21.14
1.34

1.34
1.34
1.34
1.34
214
1.27

207
+28
200
064
085
+00

CMEAN=
CHEAN
CHEAN®
CHEANS
CHEAN=
CHEANS

CMEAN=
CMEAN=

CMEAN®

CHE ANS
CMEAN=
CHEAN=
CKEAN=
CMEAN®
CMEANS

CMEANS
CMEAN®
CMEANS
CMEAN=
CMEAN=
CAEANS

CMEAN=
CMEAN®
CMEAN®
CMEAN=
CHEANS
CMEAN=

il

215 COEVe
1,70 CDEV=
4.45 COEVE

045 CDEVE
75.85 COEVs
7.45 COEVe

el SCOEV=
1.70CDEV=
4GSCUEVS
+45C0EVE
59.30 CDEV=
11.95 COEV=

1.82 CDEV=
1.70 CDOEV=
4 645C0EVE

#GSCOEVE
72.85 COEV=
9.00 CDEVe

+ 15CDEV=
1.70C0EV=
4 GS5CDE VS

«G5CDEVE

69.45 COEVe
12.50 COEV=

4.47 COEVe
G15 COEV=
4.45 CDEV=
32.95 COEV=
43.97 COEV=
7.47 COEVE

200
+00
200

1.06
07

200

00

200
3.54
349

2.37
200
200
200
35
57

200
00
00
«00
3.46
1.56

o71
2.84
+00

4.52
297


Far ee en a el oh ed ae

EUKTTTTIAATITATTULITITALILIATITIATAT IAAT AITIAARATITIITITAITIAIT
Peer ee eee ee 2 ee Serene n een ener ee gene gor ek eked eieleleleteed ded

SORTED DATA 3

1 125 1.4 -999.9 1.6 20 9.0 6.1
1 125 265 1.7 3.0 705 12.6 11.8
1 125 204 164 2.6 7.8 12.8 10.8
0 oOo 1.0 8.0 8.6 17.1 5.9 37.9
Cn) 1.3 7.0 10.1 19.9 2.4 26.3
1 0 1.1 13.3 9.9 20.1 5.9 38.4
1 0 1 22.3 14.0 30.9 12.0 42.7
1 0 0 18.6 11.9 25.8 11.9 41.8
1 0 2.3 19.2 8.5 22.6 11.7 43.9
1 0 2.3 19.2 8.0 21.8 22.3 45.7
0 2 1.5 7.0 10.1 13.0 20 69.8
o 2 2.4 11.8 10.1 15.8 16.7 56.4
o 1 264 10.6 9.9 14.8 16.8 55.9
1 1 -999.9 9.0 8.7 19.0 2.8 31.8
1 ou 1.7 10.8 lle 23.7 4.0 37.5
o 865 2.0 14.9 15.0 24.9 1703 56.7
0 5 -999.9 12.9 15.0 24.8 25.8 55.6
1 5 -999.9 7.0 8.4 16.9 2.8 29.0
1 5 1.5 5.3 8.6 15.8 3.3 27.3
Oo 25 3.3 0 +0 20 A) 20
0 25 3.8 27.0 31.4 22.4 66.2 68.6
0 25 346 25.0 28.2 21.4 60.2 62.0
1 25 2.1 12.0 10.1 28.0 -0 37.0
1 25 -999.9 1064 0 17.4 13.4 20
1 25 364 11.5 9.5 21.6 14.8 34.6
1 25 2-8 12.6 10.1 18.9 11.9 36.0
0 50 2.7 27.9 33.9 21.0 715.8 719.8
0 50 367 31.9 40.8 23.9 82.8 19.7
1 50 1.6 0 0 0 12.0 27.9
1 50 1.6 8.0 20 19.9 10.6 19.9
1 50 2-6 10.5 9.5 21.7 15.8 35.2
1 50 2.7 10.6 10.4 21.9 15.9 35466
0 125 2.3 29.6 25.7 23.7 71.0 60.8
0 125 2.4 29.7 27.9 24.0 65.3 64.4
1 125 261 18.0 13.0 28.0 0 46.9
1 125 261 18.0 20.0 28.9 25.0 43.9
1 125 3.4 17.6 12.9 28.8 22.9 41.8
1125 3.4 17.6 13.5 27.9 19.9 43.9
0 1 -999.9 -999.9 2.7 7.9 1.6 54.3
o 1 1.4 2-0 362 8.6 10.5 99.5
0 1 1.4 2.3 2.7 0 11.9 20
o 2 2.0 2.3 4.7 8.9 9.9 66.8
oo 1.0 -999.9 3.5 7.0 9.8 6402
1 61 -999.9 8 2.4 8.0 1.6 40.5
1 #1 1.6 3.2 3.2 11.6 12.0 65.8
2 1 2.0 3.7 5.6 10.8 9.8 5706
201 2.0 326 4.8 10.8 9.8 57.8
0 5 0 364 566 11.9 3.0 69e1
0 5 99909 02 4.0 8.0 1.6 47.9
a) 2.8 3.6 5.5 10.8 12.8 70.2
06S 264 361 4.9 10.8 12.8 63.8
1 5 999.9 -999.9 2.6 8.0 9.9 43.6

72


86

MANGROVE

CADMIUM
CHROMIUK
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHRON IUN
COPPER
NICKLE
LEAD
ZINC

CADM LUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
TAIN

STEM
DAY NUMBER 0
Ts +77 NOT SIGNIFICANT

Ts 28.17
Ts +00
Ts: 81
Tz a9
DAY NUMBER
T +00
Ts -00
Ts +00
Ts 00
Ts +00
DAY NUMBER

T= 4540467

Te 4.92
Te 200
Ts 2.91

DAY NUMBER
Ts 1.00
Ts 3.46
Ts 045

Ts 1.92

SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT

EMEAN =CHKE AN

NOT SIGNIFICANT -

5
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN =CME AN
NOT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
50
SIGNIFICAWT
EMEAN =ChE AN
EME AN =CME AN
SIGiWIFICANT
NCT SIGNIFICANT
NOT SIGNIFICANT
125
NOT SIGNIFICANT

NOT SIGNIFICANT
NOT SIGNIFICANT
NGT SIGNIFICANT

EXPERIMENT NO=1

EMEAN-CME AN =
EriE AN~CME AN=
EHEAN-CHE ON=
EME AN-CME AN =

EME ANTCME aN =

EME AN-CHE AN =
ENEAN@CME AN =

EMEAN-CME AN=
EAE AN-CME ONS
EME AN-CME AN=

EWE AN=<CME ANS
EME AW-CHE AN=
EME AN-CME AN =
EME AN-CME AN=
EME AN-CME AN
EME AGd~CME AN=

EME AN-CME AN =
EWE AN-CME ANS

-.03

+00
220

232

+20
73.10

-7.20
715.75
27.80

-.60

1.35
50
1.45

DEG=
DEG=
DEG=
DEG=

DEG:

DEG=
DEG=

DEG=
DEG=
DEF

DEG:
DEG=
DEG=
DEG=
DEG=
DEG=

DEG=
DEG=

Ny ©8092 Cfo Bw LYULY

DP NAN

nN

EMEANS
EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEAN=

EMEANS
EMEAN=
EMEAN=
EMEAN=
ENEAN=
EMEAN=

ENEAN=
EMEAN=
EMEAN=
EMEAN=
EREANS
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEAN®
EMEAN=
EMEAN=

230
3.80
44S
2.00

225
5-95

1.20
1.70
445
2.40

225
3.80

~80
1.70
4645
2.25
4.00
2.95

1.00
1.70
4.45
3.70
4.00
5.65

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
tDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

200 CNEAN=
+14 CMEAN#
00 CHEAN=
228 CHEAN=
+28 = CHEAN=
235 -CMEANS

00 CMEAN=
200 CHEAN=
-00 CMEAN=
200 CMEAN=
«00 CHEAN=
200 CHEAN=

700 CMEAN=
CMEAN=
CMEAN=
CHEAN=
CMEAN=
CMEAN=

CHEAN=
CMEAN=
CHEAN=
CMEANS
CMEAN®
2.06 CHEAN=

7

+33 CDEV=
1.70 COEVe
4.65 COEV=
2.20 CDEVe
+25C0EV=
5.63 CDEV=

1.00 CDEV=
4.80 CDEV=
4.45CDEV=
9.60 COEV=
16.00 CDEV=
31.60 COEV=

1.40 CDEV=
1.70CDEV=
4eG5CDEV=
1.70 CDEV=
+25 COEV=
5.75 CDEV=

1.05 CDOEV=
1.70CDEV=
Go45CDEVe
2.35 COEV=
3.50 CDEV=
4.20 CDEV=

06
00

226
40
200

whe


SOIEANAAAEMNDODOANNOCONEMROARDOODNNMNNANN DORN ANME ROT DONMOOD
ee ee eee eee eee eee te toe eee e ere eer eee eerste eee eee eres e ete s
AMNOTBODBEER OAM NODAOR EP ON RANA COGRODAMNON®ARONOMRONMHNKH STS
atte att NNN NNR te ANN NN NAR ANNE OME DON SN

Nt kt ak ont mt ead Ot mt et

NUARONAA AA A®OOORAHROBDONDNNERAAOMANMOOENMORNOFODO
coe cere eee steerer reese Ce er ee

NOM OMLVOETTHHDOMNODIUNE TUNE TONE MA CONTOMNEONODRDOAMOOND
Le rer a Oe SD SO
OONANANNANNNE SHAAN OM DORADO ETSI FANMAONN OR OOM EN AO Mantes
AAONN Nt ot At dda tt tt tod

7

%

RISE VON tH OgvatvoMoT om Ms ERM ooan DCO NOVM QOAMNOBOAAKEH EAH
a er re ee) oe eee eee eee
DODAALKEKDDRRIRDODOD PARESASHOKSOHSSSDERAAMAMANTOTE OREM
~ Attn co TNA NOTTODVORN
«

a
ANODE AODAON ATOR MN TOAE ME ENMMONVNOLHVDOMNTNAIMN MOOR ANS DOMODAM
Ce ar ar ae SS DS Se
NRA AAA AMAM SCORE ERMAN ERMOOCOM ROSEMAN INANE EOORORE
NNN Aa et tt amo

<a
ra
ZlLITQ Isis ssNNnowowNNooMasamseRonomoNnomsoDaANMyMoNnanvane
OQeeerecrcee eres e eer eee oreo ee eeereerevreeeeee ee oot eee ee eee
- Oo - as AGAMA ARAN ENN AME OME
Q ao
w nn
- te
a
oa
ANNHODODOCOCOHNNNNHNODONHNNNHOCODDCONONNNNHOOORANNNNOONNHNHOOO

WWMNMAUANN ANA DAKAMAN aaAN NNADN DON

ee ekekekel A atte -

AADQOAHADOO MM MMM MOODH MOO OM MH HOO CH HH MM MHOMNORHOHOHOMHOM MAO
NUNN NNN NNN NNN NNN NNN NN NNN NNN
hh OO
RBRERLRERERTRET TERETE E TEES LT ET TELAT ATTA TA*SASASSSSSSSFFTEZTAZS

83


Ts

Figure 31.

CU THALASSIiA ROOTS— ©&xP. iy

= F584
a
QQ
C |
a
Y owed
WJ
re
~ |
Cc
Wie |
a
s é & au
Ss
CRYS

Mean net uptake of Cu between experimental and control, Thalassia roots - Experiment I.


NI MANGROVE ROOT—- ©&xXP 1

-s4

MEAN <E-C) / PPM

CDAYS

NI THALASS!|IA ROOTS—- ©&XxP 1

=

Qa

a

<

Y ied

luo

z |

we |
a
5 é B Fy
CAYS

Figure 27. Mean net uptake of Ni between experimental and control,
Thalassia roots and mangrove roots - Experiment I.

47


Snails, clams, oysters - shell and internal organs.

Fouling organisms - total mass scraped from glass collecting
plates (23 x 30 cm). In Experiment I, all plates were sampled
at each sampling time. In Experiment II, cumulative results
were obtained by sampling plates which had remained in the
tanks for 25, 50, 85, 125 days after addition of sludge.

Thalassia - Roots and leaves. The roots and rhizomes were
combined and are referred to as roots.

Mangroves - The lower 3.8 cm of the hypocotyl not including
the roots. This is referred to in this paper as the bottom.
Growing stem includes all of the primary meristamatic tissue
from the hypocotyl excluding the leaves and the roots/rhizomes.

After dissection into these various parts, the wet weight
was obtained. The samples were dried in covered, cleaned Pyrex
beakers at 105°C. The dried sample was weighed, then homogenized
using a ceramic mortar and pestle.

TABLE 3. RESULTS IN ug o! DRY WEIGHT FOR DETERMINATION OF METALS IN NBS
BOVINE LIVER AND ORCHARD LEAF STANDARDS. THE RESULTS FOR EXPERIMENT
I AND II ARE SHOWN WITH STATISTICS AND OUR COMPUTED MINIMUM DE-
TECTABLE CONCENTRATION (MDC) .

EXPERIMENT I
Bovine Liver

ca cu cr Pb Zn Ni
MDC 0.3 8.9 3.4 0.5 2.8 0.9
x 0.56 177 <MDC 2.5 132 -
sD 0.26 20.2 96 31.7
N 9 10 10 12

EXPERIMENT II

ca cu Cr Pb Zn Ni
x 0.55 187.4 <MDC <MDC 131.7 1.0
sD 0.45 15.50 17.59 0.68
N 49 41 53 48
NBS values 0.27 + .04 193 + 10 - 0.3440.08 130+ 10

EXPERIMENT I
Orchard Leaves

x <MDC 12.3 2.6 44.8 26.1 3.9
sD 3.9 0.63 2.82 5.4 0.6
N 11 6 6 11 3

EXPERIMENT II

Orchard Leaves
x 0.66 a) - 45.0 25.6 4.0
sD 0.34 1.5 3.6 7.64 1.3
N 17 20 23 26 2.

+ 0.2

NBS values 0.11 .02 1247 (2-30 45 43° 2543 T.3


CR THALASS: A LEARVES— EXP i

MEAN (E-C> /PPM
N
Q

i] c o —
Ss Ss uu
&
CRYS
cR URCHIN GUT- EXP '
= 2a 4
a
o
S 4
a
Yo ose
ir)
v
= 4
font
w a 4
; T t r 4
s a s u
&
CAYS

Figure 18. Mean net uptake of Cr between experimental and control,
Thalassia leaves and urchin guts - Experiment I.

38


Nt THALASS |! LEAVES—- EXP.

= we
x |
a
N
a |
y 2a 4
ta
-
z 4
Cc
i 2 4
cs T T T ——+—
& 5 Fe
CDRAYS
Ni URCHIN GUT- EXP. 1
3a 4
=
ao 4
a
Ne
2e 4
a
VV
Y 4
Ls)
- 1a 4
Zz
fom 4
jr
Zea J
T — T T T 1
s & a uy
s
DAYS

Figure 22. Mean net uptake of Ni between experimental and control,
Thalassia leaves and urchin guts - Experiment I.

42


MONNAKROLNVAKRNE OTE DONONNOBGONOMMALTNRODRONARNOTE OCHROMBAOMIN
cee eee Sn Se ere ee

TO aM orem MMOL ILM NTA MMAMADDOMAADOKRTENEOMOBMNARAOGAVOR

g aAANN Ht at At tt
'
TVATM OTR YVOCORSQNAAMNDMNOAHMOOAMODAANHOOMTOTFORONAAMANDHOM ANNO
CC a TS er ee
VIA RON Se gomMaATeNNANaHaAMMSSM SSN AME ANAS ONAN ONAN TAMOR
Trang nndagsgra tte trae e reese = a aad aa a
NARA DADOOHABRAKRADENDDRDODNDTMOROAHROOTOGCLOCOORANDOOhRO000
see eee we tee ee ee ee ee ee wee eee ee oo meee eee woo r ee eeoeseereeED
MAUDRHAMA FMHAAAMNADMNSOONMAMNGAMOBNDAMNMHDO DHOHNDDOO BAD
DAAAMAMA MOAAAMAM MAMMA MAMMA MO AM tte ett ttt tet ae «
To fF ad
’

MOON MN OYMANTABSVHEVAIMNHDPONNHNHBDOONE ET ROOALTNHVOQONNAROTVON A

ee ee ee er ee rr er ee ee . se eee ee
BA AAWM A CAG TADN GON OCHA GAARA NNO AMMAN NAAM ON

Nn
TAAAMMYADMAHAAANDSOAN SARL ORE AROMAHATAAMNNHANNOAAMMOOONA ST
CC ir ee D oe ee ee ee Se
HAARANTMANTRAANAMAANAMMMMANEMAT DAR AH no BRANNAN At

aa on DAD ano BRR AH aa no
Oa o DAA ao DAR 2D aa ao
ta 1 prs v4 vod tt tt vt

=

-

TOFARMNROFMAANMOORNOSTEMMAMFANMNANHOTMFSNETMNMNDODAMNR ST FOORR

Qeeeeseesseceesseveceer et eee eee e eset eoeeeeee eee eee eee ee ee
MMAAAMOMATOMNABAMNMNKHN MMT FNM MOM TN AANA DADAM ANNA ttt NNN ON

a no a no n on

w aa an no n ow

- tt ' v4 1 '

7

o

MMNNMMAM NN ANMNNNNNODCODONNHNNHNOOCOR AA ANNNHNNNMNNL NOCOBDOONNM

NNANNN NANI ANN NINN
Aaa’

COM AAHODVOOM HA ANODOOCOM HOODOO RN MHOORHOONAODOOHAOOHAHOOCOORNAOO
okoketeketakeahakakelaltinkelateaheaReKeKeReRelto ko Rake koko ReteRoRoRo Reto ko Rot Rok koto fok kotor torre)
MANNMNNNHNHNANNNMNNNNNNHNNNHNNHNHNADHADHHADGAMHAHAANMHOMOMMHANMHHHOOHO
VVEESEVUVVVEVUVU VY VYEYEUYYYLYUYULYUUYVITTFIIEFOCLTELTILC STL CLLTrTeErTIcTrTrrz.e

71


APPENDIX D
Wet/Dry Ratios
Wet/dry ratios are shown for each sample and tissue type
for Experiments I and II. These values may be multiplied by

the metal concentration (ug/g dry weight) values to obtain
concentrations in pg wet weight.

133


92T

SEDIMENT
DAY
CADRIUM
CHROMIUM T=
COPPER
NICKLE Ts
LEAD Ts
ZINC Ts
DAY
CADM LUM
CHROMIUM T=
COPPER
WICKLE Te
LEAD Te
ZINC Ts
DAY
CACMIUM T=
CHROMIUM T=
COPPER
NICKLE Ts
LEAD =
ZINC Ts
DAY
CADMIUM
CHRGMIUM T=
COPPER Te
NICKLE Te
LEAD Te
ZINC Te
DAY
CADMIUM T=
CHROMIUM Ts
COPPER Te
NICKLE Te
LEAD Te
ZINC Ts
DAY
CADMIUK
CHROMIUM T=
COPPER
NICKLE Ts
LEAD Te
ZINC Ts
DAY
CADMIUM
CHROMIUM T=
COPPER
NICKLE Te
LEAD Ts
ZINC Ts

NUMBER
1.46

9.50
8.00
240
NUMBER

7.18

18
1.57
1.38

NUMBER

238

1.36

2.54
1.85
+14
NUMBER

2.54
2.56
8.76
08
2.05
NUMEER
1.00
290
co
1.71
1.82
3.13
NUMBER

33

290
1.30
265
NUMBER

lll
2.96

5.30
4.30

EXPERIMENT NOS2

EMEAN =CMEAN

NOT

SIGNIFICANT

EMEAN ®CMEAN
SIGNIFICANT
SIGNIFICANT

NOT
1

SIGNIFICANT

EMEANSCMEAN
SIGNIFICANT
EMEAN=CMEAN

WOT
NOT
NOT

Ss
NOT
NoT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

SIGNIFICANT
SIGNIFICANT

ENEAN =CHE AN

NOT
NOT
NOT

25

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

EMEAN=CMEAN

NOT
NOT

SIGNIFICANT
SIGNIFICANT

SIGNIFICANT

NOT
NOT

50
NOT
WOT
NOT
NOT
NOT

SIGNIFICANT
85

SIGNIFICANT
SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

"EMEANSCKEAN

NOT

SIGNIFICANT

EMEAN =CKEAN

NOT
NOT
NOT
125

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

EMEAN=CMEAN

not

SIGNIFICANT

EMEAN =CMEAN
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

EME AN-CME AN®

EME AN-CME AN=
EME AN-CME AN®
EME AN-CHEAN®

EME AN-CHEAN®

EME AN-CME ANS
EAE AN-CAE AN®
EMEAN@CMEAN®

EME AN-CMEAN®
EMEAN-CMEAN®

EME An-CME AN =
EME AN~CME ANS
EME AN-CME ANS

ENE AN-CME AN
EME AW-CME AN=
EME AW-CHE AN=
EME AN-CMEAN®
EME AN-CME AN

EME Aiv-CMt AN
EME Aii~CME ANS
EME AN-CME AN®
EME AN-CHE AN®
EME AW-CME AN=
ENE AN-CME ANS

EME AN-CMEAN®
EME AN-CME AN®
EME AN-CME AN
EME AN-CME AN®
EME AN-CME AN®
EME AN-CME AN=

EME AN~CME AN
EME AN-CME AN®

-.50
+30
633

2610

DEG=

DEG=
DEGs
DEG=

DEG

DEGs
DEG=
DEG

DEGs
VEG

DEG
DEu=
DEG=

DEG=
DEGs
DEG=
DEG=
LEG=

DEGs
DEG
DEG=
DEG=
DEG=
DEG

OEGe
DEG=
DEG=
DEG=
LEG:
DEG=

DEG=
DEG=

Pep 5

Serene peer

bpp oe

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN=

EMEAN®
EMEANE
EMEAN®
EMEAN=
EMEAN=
EMEAN®

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEANS
EMEANS
EMEAN®
EMEANS
EMEANS
EMEAN®

EMEAN®
EMEANS
EMEAN®
EMEANS
EMEAN=
EMEAN®

EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

260

14.50

+00
oh2
2.92
064

7 6&5
1.78

1.78
2.40

2240
40
46
025

225
1.27

1.27
047
31
bh

CHEAN=
CHEAN®
CHEAN=
CMEAN=
CMEAN®
CHEAN®

CHEAN=
CMEAN®
CHEAN®
CMEAN=
CHEANS
CMHEAN=

CHEAN=
CMEAN=
CHEAN=
CHEAN=
CMEAN=
CMEAN=

CMEANS
CHEAN=
CHEAN=
CMEAN=
CMEAN=
CHEAN=

CMEAN=
CHEAN=
CHEAN®
CMEAN=
CMEAN=
CMEAN=

CMEAN=
CMEAN=
CHEAN=
CHEAN=
CMHEAN=
CMEAN®

CHEAN=
CMEAN®
CMEAN=
CMEAN=
CMEANS
CHEAN®

36

o6O0CDEV=
7.17 COEV=
4.G5CDEV=
9.37 CDEV=
7.73 CDEV=
16.20 COEV=

STCDEV=

7 CDEV®
oG5COEV=

6.30 CDEV=

6.67

COEVe

15.93 CDEV=

28
6.73

97
6.53
13.80

COEVe
COEV=

SCDEV=

COEV=
CDEV=
CDEV=

oGICDEV=

7.87
9.60
8.87
6.90
15.90

240
6.57
5.93
7.30
4.90

12.07

CDEVe
CDEV=
CDEV=
CDEV=
COEV=

CDEV=
COEV=
COEV=
COEV=
CDEV=
CDEV=

«20CDEV=

42.67

COEV=

4 4S5CDEVE

8.33
6677
15.47

COEV=
CDEVe
COEV=

23 TCOEV=

52.00

CDEV=

445 CDEV=

10.37
7el7
15.43

COEV=
COEV=
CDEV=

59


Sewage Sludge

Activated sludge was shipped from New York City in 208 & poly-
ethylene lined drums. The sludge was allowed to settle and the
overlying liquid decanted. In general, sludge is 5% solids. The
black, "oily" sludge slurry remaining was mixed in a large fiber-
glas tank. The sludge was added to a depth of 7.6 cm in an area
1m wide by 2.3 m long (0.1 m3) at the incurrent end of the
Experimental tank which had previously been sealed off from the
rest of the tank. A PVC frame with small mesh screen attached
was placed over the sludge to prevent direct contamination of the
organisms. A duplicate screen was placed in the control tank.

After 24 hrs, when the sludge had settled, the seal was removed

and Experiment I was begun. At days 1, 5, 25, 50, 85 (Experiment II
only) and 125, after the sludge was added to the Experimental tank,
samples of organisms, sediment, sludge (day 1, 125 EXPI) and water
were taken from both tanks. A presample was taken prior to addition
of any sludge, in this study it is referred to as day 0.

Sampling Organisms
The initial stocking quantity or density for the biological
organisms, used in both Experiment I and II, is shown in Table 1.

TABLE 1, PHYSICAL DIMENSIONS OF ORGANISMS WITH NUMBER OF TOTAL ORGANISMS
STOCKED PER TANK AND NUMBER OF ORGANISHS SAMPLED PER TANK PER DAY

EXPERIMENT I

Experimental Tank Control Tank
Low H___fistocked #/Sample* L w H #stocked #/Sample*
Clams 15.1 104 10, 39.3 36-8 17.2 104
3.2 20 7.4 25.7 17.2
Sea Cucumbers X 29.2 110 Q 35.9 29.4 101
. 1 8.0
Snails 124
Oysters
Urchins
- = ee H
Clams x 40 34.6 15.8 39.3 36.1 15.4
S.D_7.5 6.3 3.4 _ 2.5 7.4 3.2 ; ee
Sea Cucumbers x 44.6 35.8 $2 5 44.5 35.2 52 5
S.D, 17.8 10.4 13.4 10.4 7 _
Snails & V1.6 180 20 1S.5 11.5 180 207 *
9-7 — ae ee
Oysters a 132 10 38.6 131 10
Do 9.5 _.
Urchins & 45.8 9.2 57 5 47.4 29.7 37 S
s.D 5.2 4.0 _ $.7_ 3.5 _

*Upper values are for days 0 through 50, Experiment I, lower values for
day 125, kumbers refer to number of organisms in cach pooled sample.

**Snails net taken on day 125 control as none loft.

5


82T

THALASSL RGOT EXPERIMENT NO=2
DAY NUMBER 0
CADMIUM NO EXPERMENT MEAN= 1.20 DEVIATION=
CHROMIUM NO EXPERMENT MEAN= 17.80 DEVIATIGN=
COPPER NO EXPERMENT MEAN= 4.45 DEVIATION=
NICKLE NO EXPERMENT MEAN= 26.47 OEVIATIUN®
LEAD NO EXPERMENT NEAN= 10.13 DEVIATION
ZING NO EXPERMENT MEANS 34.93 DEVIATION=
DAY NUMBER 1
CADMIUM T= 1.77 NCT SIGNIFICANT EMEAN-CMEAN®
CHROMIUK T= 1.80 NOT SIGNIFICANT EMEAN-CHEAN®
COPPER Ts 244 NOT SIGNIFICANT EMEAN-CME AN®
NICKLE Ts 2.69 NOT SIGNIFICANT EMEAN-CMEAN=
LEAD Ts 14.63 SIGNIFICANT EME Aiv-CME AN =
ZINC Te 03 NOT SIGNIFICANT EME AN-CME AN®
DAY NUMBER 5
CACHIUM T= +50 NOT SIGNIFICANT EME AN-CMEAN=
CHROFIUM T= 29.57 SIGNIFICANT EME AN-CME AN=
CUPPER Ts 15.06 SIGNIFICANT EME AN-CMEAN®
NICKLE Ts 24.77) SIGNIFICANT EME AN-CME AN=
LEAD Te 8.57 SIGNIFICANT EME AN-CMEAN®
ZINC T= 8.69 SIGNIFICANT EME AN-CHE ANS
DAY NUMBER 25
CAOMNIUM T= 9.19 SIGNIFICANT EME AN-CME AN
CHROMIUM T= 1.71 NCT SIGNIFICANT EME AN-CMEAN®
CUPPER T= 2.44 WOT SIGNIFICANT EMEANW-CHt AN®
NICKLE T= 2-76 NCT SIGNIFICANT EMEAN-CME AN®
LEAD Ts 204 NOT SIGNIFICANT EME AN-CMEAN®
ZINC Ts 5.75 SIGNIFICANT EME AN-CME AN®
DAY NUMBER 50 .
CADMIUM T= 1.07 NOT SIGNIFICANT EMEAN-CMEAN®
CHROMIUM T= 6-71 SIGHIFICANT EME AN-CRE ANS
COPPER T= 9.41 SIGNIFICANT EME AN-CMEAN®
NICKLE T= 1.98 NOT SIGNIFICANT EMEAN=CME ANS
LEAD T= 7.87) SIGNIFICANT EME AN-CME AN®
ZINC T= 4.53 SIGNIFICANT EMEAN-CME AN®
DAY NUMBER 85
CADMIUK T= 3.13 SIGNIFICANT EWE Aw-CME AN®
CHROMIUM T= 7-43 SIGNIFICANT EME AN-CHE AN®
COPPER Te +53 NOT SIGNIFICANT EMEAN-CMEAN®
NICKLE Ts 1.72 NOT SIGNIFICANT EMEAN-CHE AN®
LEAD T= 1.72 whT SIGNIFICANT EME AN-CME AN=
ZIKC T= 2.31 NOT SIGNIFICANT EME AW-CMEAN®
DAY NUMBER 125 7
CADMIUK T= 2.24 NOT SIGNIFICANT EMEAN-CHE AN®
CHROMIUM T= 4.55 SIGNIFICANT EME AN-CME AN®
COPPER Te 278 NOT SIGNIFICANT EMEAN-CME AN®
NICKLE T= 1.92 NGT SIGNIFICANT ENEAN-CMEAN®
LEAD Ts 17605 SIGNIFICANT EME AN-CHE AN®

ZINC

Te 14.49

SIGNIFICANT EME AN-CME AN =

+10 NUM=
1.15 NUM=
+00 NUM®
+98 NUM®
1.53 NUM®
NUM®

weUvUe

DEG=
OEGs:
Oboe
DEG=
DEGs
DEGs

OEGs
DEGe
DEG=
DEG=
DEGe
DEG=

DEG=
OEG=
DEGs
OEG=
DEGs
DEGs

bEos
OEGe
DEGs
DeGe
DEG
OEG=

vEG=

CEG
7

DEGs
OEG=
DEG=

DEG=
DEG=
DEG=
vEG=
DEGs
GEGe

PREP ESR eEEess

Brees

appeee

PRUE SE SPEED

EMEAN®=
EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN=
EMEAN®
EMEAN®
EMEAN=
EMEAN®
EMEAN®

EMEANS
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEANS
EMEAN®
EMEAN®=
EMEAN®
EMEAN®
EMEAN®

EMEAN=
EMEAN=
EMEANS
EMEAN®
EMEAN®
EMEAN=

EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEVe
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
eDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
eDEV=

206
1.36
1.30
1.30

274
2.25

+00
2.20
1.23
2.61
1.52
290

+06
2.06

38

CMHEAN®
CHEAN=
CHEAN®
CHEAN®
CHEAN=
CMEAN=

CHEAN=
CHEAN=
CMEAN=
CHEAN=
CHEAN=
CHEAN=

CHEAN=
CHEAN=
CHEAN=
CHEAN=
CMEAN=
CHEAN=

CHEAN=
CHEAN=
CHEANS
CHEANS
CHEAN=
CHEAN=

CHEAN=
CHEAN=
CHEAN=
CMEAN=
CMEAN=
CMEAN=

CMEANS
CHEAN=
CHEAN®
CMEAN®
CHEAN®
CMEAN®

38

COEVe
CDEV=
COEV=
COEV=
COEVe
COEV=

CDEV=
COEV=
COEV=
COEV=
CDEVe
CDEV=

COEV=
COEV=
CDEV=
COEVe
CDEV=
CDEV=

COEV=
CDEV=
COEV=
CDEV=
CDEV=
COEV=

CDEV=
CDEV=
COEV=
COEV=
COEV=
COEV=

COEV=
CDEVe
COEV=
COEV=
CDEV=
COEV=

1.86
5.39

212
4.23
3.54
1.98

1.38


<b THALASS (IA LEAVES—- EXP. 2

MEAN (E-C) / PPM

ay
th
ea
ez!

DAYS

CD URCHIN GUT- EXP. 2

m4

MERN <E-C)> - PPM

DRYS

Figure 26. Mean net uptake of Cd between experimental and control,
Thalassia leaves and urchin huts - Experiment II.

46


ABSTRACT

This research was initiated to determine the rates of
uptake, by a Thalassia testudinum ecosystem, of Cd, Cr, Cu, Ni,
Pb, and Zn which were leached from sewage sludge by sea water.
The experimental design used aerated flowing sea water (8.42
min 1), which passed over a 0.1 m3 bed of sewage sludge before
traversing the model ecosystem. The tanks, both control and
experimental, were 9.2 m x 0.9 m x 1.1 m in size with a volume
of 3.1 x 10% liters. Each tank contained sand to a depth of
0.5m for a total volume of 4.2 m¥. The experiment ran for 125
days from March 1975 to July 1975 and was duplicated from
December 1975 to April 1976.

The largest net uptake of metals occurred in the "fouling
organisms" where Cd, Pb, and Zn uptake closely paralleled the
net loss of metals from the sewage sludge. Thalassia leaves
showed a net uptake for Cr, Pb, Ni, and Zn for both experiments.
The urchin (Lytechinus variegatus), a herbivore on Thalassia
leaves, also demonstrated a net uptake of Cu, Cr, Pb, Zn, and
Ni in both experiments. The sea cucumber (Holothuria mexicana)
in both experiments showed a net uptake of Cr, Cu, Pb, and Zn.,
Net uptake of metals in mangroves (Rhizophora mangle) was limited
to the roots. In the first experiment Ni and Zn showed a
significant uptake whereas in the second experiment only Cr
demonstrated a net uptake. The uptake in mangrove roots appeared
to be a direct function of metal concentration in the sediment.
The lack of apparent metal uptake in the sediment, except for
Ni, Pb, and Zn in the second experiment, was probably due to
the sampling technique rather than to a lack of uptake of metals
by the sediment.

No consistent or significant metal uptake was found for the
clam (Codokia orbicularis), oyster (Crassostrea rhizophora) or
the snail (Nerita tessplata).

The lack of sufficient sample mass, for these organisms,
probably obscured any net uptake that may have occurred.

The results indicate that the dumping of sewage sludge in

coastal tropical waters can lead to the uptake and concentration
of toxic trace metals by members of a turtle grass community.

iv


PB HOLOTHURIAN GUT— EXP. 2

ae 4
=
a 4
o
S
ze 4
“a
u
i 4
WW
vo oip4d
Zz
Cc 4
Wi
ze
T \ T r

a4
Bh
a8

azt

DAYS

ZN HOLOTHURIAN GUuUTt- xP =

4
as |
i
o 4
~
is 4
A
v 4
1
w
vs |
z
£ 4
i
= -S 4
ot

a
@h
[2]

a21 4

Figure 43. Mean net uptake of Pb and Zn between experimental and

control, sea cucumber guts - Experiment II.

63


only available for two sampling times in Experiment I, so it
is difficult to determine the true instantaneous net loss of
metals from the sludge.

The original objectives for this research, as stated in the
Introduction, have been obtained. A definite net uptake was
found in members of a tropical marine community. These results
were replicated at two different seasons and the results, con-
sidering the complexity of the system, agree quite well.

Originally, the fish, Diodon holacanthus (puffer) was to
be added to the system. However, after collecting over 360
specimens we found that they were too sensitive to environmental
factors other than sewage sludge and were easily killed. There-
fore, fish were deleted from the ecosystem in order to expedite
the completion of the experiment.

19


CU HOLOTHURIAN GUT- ©&xP 2

MEAN <E-C)> /PPM

@
Bh
me

az

DAYS

NI OHOLOTHURIAN BWT- EXP. 2

MEAN <E-C> PPM

4

a
Gh
Viz]

azi

Figure 41. Mean net uptake of Cu and Ni between experimental and.
control,. sea cucumber guts - Experiment II.

61


PB MANGROVE ROOT—- EXP. 2

=

i 1a4

S

“a

oO. |

WwW

v

Zz

fe

W

=

“= a ot ot
. & & z

CARYS

PSB THALRASS!IAR ROOTS— EXP. 2

za,
= |
x
x
~ 14
A
Ys
rt)
-
wp 4
Zz
Cc
y -= 4
—1a T T T T 1
ss —
6 g 2
DAYS

Figure 34. Mean net uptake of Pb between experimental and control,
Thalassia roots and mangrove roots - Experiment II.

54


SECTION 3
RESULTS

The raw data, expressed as ym metal/g dry weight for all
samples, are found in Appendix A. The computed net uptake for
each metal is located in Appendix B and is expressed as mean
experimental results (ug/g dry weight) minus the mean control
results for each sample, with their respective standard devi-
ations, and degree of freedom. The "t" value for the null
pypothesis, Ho: = 0 (P<0. 05) where M, is experimental and

is control is “1168 shown in Appendix B. The analyses for the
nas biological standards are shown in Table 3. The results for
all fractions of dissolved and particulate trace metals in the
water for both control and experimental tanks (Experiment II)
are in Appendix C. The wet/dry weight ratios with statistics
for all the samples in Experiment I and II are in Appendix D.

The experiments were replicated, Experiment I began on
11 March 1975 and ended 15 July 1975; Experiment II began on
December 12, 1975 and ended April 26, 1976. The experimental
tank refers to the tank with the sludge added and the control
tank was identical, in experimental design, to the experimental
tank except no sludge was added.

Temperature, salinity, and oxygen

In Experiment I the temperature ranged from 21 to 28°C
with a salinity range from 32 to 34°/,,. In Experiment II the
temperature ranged from 21 to 26.5°C with a salinity range from
24 to 35°/45(%, 30°/o9, S.D. 4.2°/4,, N = 18).

Dissolved reactive phosphate (DRP)

The results are shown in Figs. 4 and 5 for control,
Experiment I and II and experimental, Experiment I and II,
respectively. The plotted points are the mean values for a
five day period.

Fouling organisms

In Experiment I, Cd, Cu, Pb, and Zn showed a significant
net uptake after 125 days (Figs. 6, 8, 9, 11), whereas in
Experiment II, Pb (Fig. 10) showed a steady uptake to the end

12


cu FOULING ORGANISMS— EXP. 1

MEAN <E-€> PPM

a4
Gh 4
267)
e214

pDAYS

cu SEWAGE-—- EXP.

mome S
zs |
zeae 4
sae 4

MERN PPM

som 4

a
Bh
C2}

e214

Figure 8. Mean net uptake of Cu between experimental and control,
fouling organisms and sewage - Experiment I.

28


NI MANGROVE .ROOT—- EXP. =

is,
=
aa 1a 4
N
“A
Y = 4
W
vv
Zz
crea 4
ld
=
-= T T T + T T T
= & z 3
CDAYS
Ni THRALASSIA ROOTS- ©xXP. 2
eo,
= 4
a
a wd
S
na
y 4
Lo 22 4
v
Zz 4
c
prt
zap
r r T T T T 1
8 é 8 y
DAYS

Figure 28. Mean net uptake of Ni between experimental and control,
Thalassia roots and mangrove roots - Experiment II.

48


TABLE 4.

CONCENTRATION OF Cd, Cr, Cu, Ni, Pb AND Zn IN ug/g
DRY WEIGHT, IN SEWAGE SLUDGE USED IN THIS EXPERI-
MENT. THE RESULTS ARE FOR EXPERIMENT I AND II.
RESULTS FOR THE SAME METALS AS DETERMINED BY SALOTTO
AND FARRELL (1971) FOR ACTIVATED SLUDGE ARE INCLUDED.

EXPERIMENT I

ca Cr Cu Pb* Ni Zn
x 109 2120. 2560. 60 319 2926
sD 6.70 115. 124.0 55 11.3 388.4
EXPERIMENT IT
ca cr cu Pb* Ni Zn
x 166. 3667 2445. 21.0 376. 2508
sD 85.5 338.5 266.4 3.29 12.7 122.1

SALOTTO AND FARRELL (1971)

ca cr cu Pb* Ni an

350 4330 1100 1500 380 3300

* Lead values are low due to digestion technique.

15


TECHNICAL REPORT DATA

(Please read lustructions on the reverse before completing)
TALPORT NO. 2 3. RECIPIENT'S ACCESSION NO,
|. EPA-600/ ae _. 4
4. TITLe AND, 5. REPORT DATE
BIOLOGICAL AVAILABILITY OF POLLUTANTS TO MARINE March 1978 issuing date
ORGANISMS 6. PERFORMING ORGANIZATION CODE
7, AUTHOAIS) Is. PERFORMING ORGANIZATION REPORT NO

John R. Montgomery, Mary Price, John Thurston, Gina
Laite de Castro, Luz Leida Cruz and Domenica deCaro Zimmerman

19. PERFORMING ORG *NIZATION NAME AND ADDRESS 10. PROGRAM ELEMENT NO.
Center for Energy and Environment Research 1BA608
Mayaguez, Puerto Rico 00708 TT. CONTRACT/GRANT NO,

TAG-D-4-0541

12. SPONSORING AGENCY NAME AND ADDRESS 13. TYPE OF REPORT AND PERIOD COVERED
Environmental Research Laboratory Final
South Ferry Road 14, SPONSORING AGENCY CODE
Narragansett, R. I. 02882 EPA/600/5

16, SUPPLEMENTARY NOTES.

16. ABSTRACT

This research was initiated to determine the rates of uptake, by a Thalassia
testudinum ecosystem, of Cd, Cr, Cu, Ni, Pb, and Zn which were leached from sewage
sludge by seawater. The experimental design used aerated flowing seawater (8.4%
Min-!), which passes over a 0.13 bed of sewage sludge before traversing the model
ecosystem. The tanks, both control and experimental, were 9.2 mx 0.9 mx1.1 min
size with a volume of 3.1 x 103 liters. Each tank contained sand to a depth of 0.5
for a total volume of 4.2 m3. The experiment ran for 125 days from March, 1975
to July, 1975 and was duplicated from December 1975 to April 1976.

17. KEY WORDS AND DOCUMENT ANALYSIS

i SCRIPTORS D.IDENTIFIERS/OPEN ENDED TERMS |c. COSATI Ficli/Group
Water Pollution Metals Uptake from 57H
Underwater Environments sewage sludge

Aquatic Animals

18. DIS THIGUTION STATEMENT 12. SECURITY CLASS (This Report) 21, NO, OF PAGES
UNCLASSIFIED 145
RELEASE TO PUBLIC 20. SECURITY CLASS (This page) 22. PRICE _
UNCLASSIFIED
EPA Form 2220-1 (3-73) 135

U.S, GOVERNMENT PRINTING OFFICE:1973 260-880/37 1-3


BBTTITISTIALTIIGILATIAIIITIITLIAILIIIATIIITITrTITrTzAITIIAIAIAITIITITIEIIIT
Ee ee ee a ete te te et ee et ted Ped ed td tet et OOOO OM ODT POT MOTDTOWOD

RNNNNN NNN N NNN NNN NN NNNNNY NN RNR NTN NNR MNT er
OMe ROD OPH HOO OP RP HOO RE H ODOR RH OO OR EEE RH OOOH HH OOO MH HOON ES

SORTED DATA

25
25
50
50
50
50
50
85
85
85
85
65
85
125
125
125
125
125

ry
nN Ny
MUU Ee ee eR OOOM

RNNND
wun

woounuUuUUNN
VUNOOOCOS

wo %
wu

125

ee ee eee ny

ee pe

4
3
8

1.1

8

1.0

9

BERS SS

8
7
7
7
7

eee eo ee we

°

°

.
ODODDOOOYWE TE RUBOUUNNWOR

FOUWSHOse SEN
ee ee or er rr re rar ry

He ODOUE
ONINOFLOUVDON OP UNO O Ie

1.5

nN

Perera rer ee Se rer Or a ar ars

eee eee

WODMNUMUTTIN NN UUUYWNWUYWE FLUSHED
PONUNUFAABOUGHE EE ENINAOTVWYUWORY

eee eee

22.9
15.0
15.5
14.2
23.6
23.8
23.9
14.0
13.1
13.2
12.8

eee

Perera aera sr ari ar aren er ar rer er ar ary

RNYNeERN
BOSKUUNFOOCOHUUE HERE AORAAOTOUMUUE

cee

RUF OOBVHROUNSIH RK SSHSTOUNUNOL ODN

- 999.9
-999.9

8.2
11.7
13.8
24.3
15.6

enone
NYWwW

me

NNW

VwWvvwwowoeof Fass sw
NOEOUVUWNIWINN NNN TNOBHOY

errr wren wren ar ner ne wer ae Sy

eee
Von
cOnuwh

we
w
.

roy

12.5
15.8
25.2
23.6

99909

13.5
14.7
12.4
10.4
10.1
8.2
6.4
8.2
49.
48.3
46.4
15.8
16.6
17.0
15.5

een

e
SMVUYVHINFTOVUNSVITUNFONY

Per ere ee eee Or ae ee

Nu

wn

ao

.
WON NANWUNUNU HE OH OON ENTS RRO

ww
oo

39.3
43.8
48.2
48.9
56.1
51.9
65.6
45.8
48.9
47.0
47.5

0
40.0
41.5
44.1
35.1
40.0
40.3
29.8
27.9
28.6
67.7
64.1
62.2
40.5
45.2
39.6
50.0


CR FOULING ORGANISMS— ©&xP 2

2eo

tee 4

MERN (E-C> , PRM
8
8

2a

e217

CR SEWAGE- Exe. 2

“smo
4
aso |
=
a
a 2sea J
z 4
Cc
Bixee |
soo
+ T T 1 1
= & 8 Fy

cCAYS

Figure 15. Mean net uptake of Cr between experimental and control,
fouling organisms and sewage - Experiment II.

35


AANA AF HMA HRA HHH HUH HDUUH UHHH HHHDHHNUNOHY

PRP rrr rrr rrr rrr er rere rere eee

Re ee ee

KPH H ODO DDO OP EH ODO OH HEE HOODOO ORB HE ROO HH OOH MOON MOOR HOO ROO

SORTED DATA

VUUYN Ee EH OOOO

NUN
WUT

TUN N
e000

125
125
125
125

VU Pe eee EB OOO

NuND
Vann

NNN RN
WUT UU

999.9
999.9
999.9
999.9
999.9
999.9
-999 9
999.9
999.9
999.9
999.9
999.9
999.9
999.9
-999.9

6

wee ee ee th ew ow
POWwOwT ec owwoorkooyeuw

ae

Oo
NOOWN KEP WWN RK UDNNUWE ND

eee

Pair era are ie ee er ee

~

ore

'
.
ONNMOWOMK FOSTTVIBSOUTIIUVOBIOUIS

ee ee eer or

CMBBIOMONVIUTFWONCODOWVDDCOGC9Q0COCCO00

we

31.
23.6

23.9

76

eee ee eee

~
HU Oer INP OONOONES

~
NANMDODKODOODCDOGCSDOO

eee ee we wee

ray
IM OOO

12.9

42.7

tee ee ee
CONE SUH
eee eee eee

epwwoovvocrcoev

er
uw

15.9

CWO WOW
eeee

31.8

19.7

21.8
24.8
20.3
20.0
20.6
21.9
20.7
20.7
17.1
19.2
20.7
21.1
20.6
18.7
23.5
18.8
18.0
186.8
19.7
16.8
19.4
17.2
24.3
19.2
47.3
30.1
32.5
53.9
45.8
48.7
52.8
4364
39.4
37.8
39.3

-0
47.2
45.9
44.8
37.0
34.4

30.7

0
27.4
27.5
29.8
26.1
31.7

20
34.6


€s

Pigure 33.

PB THALASS!:A ROOTS—- &XxP 1

4
ue 4
= 4
a
a se
x
A 4
Y zed
uw
v 4
zie
jo
Wi 4
=
2 4
‘i a a a
s 6 8 3
DAYS

Mean net uptake of Pb between experimental and control, Thalassia roots - E

yperirent I.


SCCCECLSSCCECCCCE HAHAHA At ttt
et et eet tat tet tat oat Oot Ot ts ot oe IO FO WW WDD DDD DDD DDD DD HDD DDR HD RDDVWO CKO rR
POP a ke te teh tek ee et ee ee tek ee bt be Ded be ee tee Dba Bat tad ba a ba tt tat ba Pot tt att tat bt tt te bt tt
RH ROORP HOOP HOD OCOHP ROO OOP ERE H OOO ORE HH OOCOOP RE HOOH HEH HOOOF

SQRTED OATA

25
50
50
50
50
50
50

nN
MUNN ee ee OO

nn
ww

NNN
VM

wu
ooo

ww
LMU Ue eee OOOO OCOGCOO

eee eee eee oe

v
eo
o
NWEWOE NANNY NNNOUE HENNE EUW

se ee

ONWVOODRNUDHKKOOWRFOOWVOWN

eee

WODRITH EO

.
ree

-

ee cr on

ce ee eee

Ve ODDWNHPWONH

ee eee

SR oOR Rens
coe ee

a
nN
.
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eta
o

ecee

~o
.
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75.9
717.2
59.0
46.7
37.3
44.44

-99909
-999.9

46.1
40.9
29.9
2768
30.2
30.4
43.6
43.8
48.1
50.3
404
45.8
43.7
40.8
46.9
50.4
43.7
42.6
37.5
33.8

31.4
46.2
46.9
56.5
51.9
33.8
34.9
16.8
19.7
13.9
15.3
36.9
35.9
21.4
21.4
27.0
23.3
39.8
39.7
27.7
31.4
2329

94.7

100.0

91.3
16.7

0
19.8
15.5
15.5
28.4
28.9
20.0
21.8
20.1
21.2
31.7
27.9
24.0
25.6
15.8

0

19.9
22.0
2407
21.8
19.9
24.7
24.8
17.8
21.6
39.0
41.2
4906
48.0
12.3
12.1
13.1
15.1
12.9
13.4
19.9
16.0
16.1
19.46

20
18.7
29.0
27.4
20.9
16.6
16.2


aan

CLAM

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
LINC

CADM TUM
CHROMIUM
COPPER
NICKLE
LEAD
TINC

CADN TUN,
CHRUMNIUM
COPPER
NICKLE
LEAD
ZINC

CADN TUM
CHRGM UM
COPPER
NICKLE
LEAD
ZINC

CACM IUK
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM IUK
CHROMIUM
COPPER
NICKLE
LEAD
LIN

INTERNAL EXPER

DAY NUMBER 0

NO
NO
NU
No
NO
NO

EXPERMENT MEAN= 2.1
EXPERMENT NEAN= 3.7
EXPERMENT MEAN= 2.
EXPERMENT NEAN= 6
EXPERMENT MEANS 3.7
EXPERMENT HEAN= = 11947

DAY NUMBER 1

Te

Te

+35 wOT SIGNIFICANT
ENE AW SCKE AN
10.56 SIGNIFICANT
1.44 NOT SIGNIFICANT
263: NGT SIGNIFICANT

14.29) SIGNIFICANT
DAY NUMBER 5
Ts 1.26 NET STONIFICANT
T= ell NCT SIGNIFICANT
Ts 2.50 NCT SIGNIFICANT

1.80 NOT SIGNIFICANT
269 NGT SIGNIFICANT
1.74 NOT SIGNIFICANT

DAY NUMBER 25

Ts

214 WOT STGNTFI CANT
4.00  SIGHIFICANT
7.48 SIGNIFICAWT
3464  SIGWIFICANT
225 NCT SIGNIFICANT
244 NGT STONIFICANT

DAY NUMBER 50
Ts

Ts

4024 SIGwIF ICANT
1.32 NGT SIGNIFICANT
7.52 SIGwIFICANT
1.73 WOT SIGNIFICANT
1.91 NGT SIGNIFICANT
5.41 SIGHIFICANT

DAY NUMBER 85

Te
Ts
Ts
Ts
Te
Te

5.50 SIGNIFICANT
2.18 NOT SIGNIFICANT
+83 NOT SIGNIFICANT
3.62 SIGNIFICANT
«08 NOT SIGNIFICANT
+67 NOT SIGNIFICANT

DAY NUMBER 125

Ts
Ts
Te
Ts
Te
Te

11.02 SIGNIFICANT
1.25 NOT SIGNIFICANT
15.28 SIGWIFICANT
+59 NCT SIGNIFICANT
1.85 NOT SIGNIFICANT
7.15) SIGNIFICANT

IMENT NO=2

3 DEVIATIONS
3 DEVIATION=
DEVIAT HONK
O OEVIATIUNS
2 OEVIATIUNS
0 DEVIATIUN=

EME AN-CME AN =

EME AN-CME AN =
EME Aiv-CME AN =
EME AH-CHE AN=
EME AN-CMEAN=

EME AN~CMEAN®
EME Aiw-CAE An =
EME AN-CHE A=

EME An~Ce AN®
EME Aw-CME ANS
EME AN-CME AN=
EME AN-CME AN
EME AN-CME AN=
EME AN-CME AN=

EME AN~CMEAN=
EME AN~CME AN =
EME AN-CME AN
EME AN-CME EN=
EME AN-CME AN=
EME AN-CME ANS

ENE AW-ChE aN =
EME AN-CME An =
EME AN-~CME ANS
EME AN-CME AN =
EME AN-CME AN =
EME AN~CHE AN =

EME AN-CME AN=
EME AN-CME AN=
EME AN-CME AN=
EME AN-CME AN =
EME AN-CME AN =
EME ANRCME AN

212 NUM=
212 NUM®
1.10 NUM®
+36 NUH
3.04 WUM=
7.30 NuUMe

VeEwoue

-.03 DEG=

7-71.33 VEG*
~.30 DEG=

027 DEG=
42.87 DEG

+37 DEG
+13 LEGe
6.90 DEGs
+77 VEGe
1.13 DEGs
30.07 vEuw

+27 DEGe
80 DEG=
6.97 VEGe
77 OEGe
210 DEG
73.00 OELs

~.20 DEG
227 DEGs
6.77 DEG=
+10 DEG=
1.77 DEG=
39.23 DEG=

037 DEu=
1.03 LEGS
-1.60 DEG=
«57 DEG=
03 DEG=
2.93 DELS

«90 DEG=
1.30 DEG=
15.70 OEG=
+27 DEG=
+77 DEG=
"41.40 DEG=

peppers

PED EEE pesese

pepe as

popes

EMEAN®
EHEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN®=
EMEAN=
EMEAN=
EMEANE
EMEANS
EMEAN=

EMEAN=
EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEANE

EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=

EMEANS
EMEAN=
EMEANE
EMEAN=
EMEAN=
EMEANS

1.13
1.70
20.13
5.80
4.27
150.50

1.33
3.47
23.07
5-20
6.37
119.47

1.83
4.00
20.20
5.90
5-23
90.00

1.57
4ell
20.03
3.90
5.23
100.67

1.40
7.13
22.20
6.37
5.37
106.20

2.40
4.00
27.43
5.87
4.10
90077

EDEV=
EDEV=
EDEVe
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EOEV=
cOEV=
EDEV=
EOEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

206
206
+76
220
264
2.60

025
1.59
4645

53

98

12.23

21
35
1.25
20

40
9.36

206
«06
215
210
1.50
11.63

210
55
1.ll
1

55
4.23

+10
52
2246
59
210

CHEAN®
CMEAN=
CHEAN=
CHEAN=
CHEAN=
CMEAN=

CHEAN=
CHEAN=
CMEAN=
CHEAN=
CHEAN=
CHEAN=

CHEAN=
CHEAN=
CHEAN=
CHEAN=
CHEAN=
CHEAN=

CMEAN=
CHEAN=
CHEAN=
CMEANS
CMEAN=
CHEANS

CMEANS
CMEAN=
CHEAN=
CHEAN=
CHEAN=
CMEAN=

CHEAN=
CMEAN=
CMEAN=
CHEAN=
CMEAN=

212 CMEANS

22

217

COEV=

1.70C0EV=

27.47
6.10
4.00

107.63

1.70
3.33
16.17
4043
5-23
89.40

1.57
4.80
13.23
6.67
5.13
93.00

1.77
3.90
13.27
3.80
3.47
61.43

1.03
6410
23.80
5.80
5.33
103.27

1.50
2.70
11.73
5.60
3.33
132.17

COEV=
COEV=
CDEV=
COEV=

COEV=
CDEV=
CDEV=
COEV=
CDEV=
CDEV=

COEV=
CDEV=
CDEV=
CDEV=
COEV=
CDEV=

CDEV=
COEV=
COEV=
COEV=
COEV=
CDEV=

CDEV=
COEV=
CDEV=
COEV=
COEV=
COEV=

COEV=
COEV=
COEV=
COEV=
COEV=
COEV=

00
93
30
336
4.50

246
1.46
1.74

51
2.68

27.37

206

7.01

1.37
+90
258

4.15

06
61
3.14
217
45
6.30

210
1.73
1.01

252

7
7295


STT

HOLOTHUR

CADMIUM
CHROMIUH
COPPER
WICKLE
LEAD
TING

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
1IKC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
WICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE

CADMIUM
CHROMIUK
COPPER
NICKLE
LEAD
qin

DAY NUMBER

NO EXPERMENT MEANS
NO EXPERMENT MEAN=
WO EXPERMENT MEAN=
NO EXPERMENT HEAN=
NO EXPERMENT MEAN=
NO EXPERMENT HEAN=

DAY NUMBER
cy

Ts
Ts 2.00
Te 36.55
1s 22
Ts 4.64
DAY NUPBER
Ts 49
Ts 5643
7.61
16.07
1.76
2.56
DAY NUKBER
ia 1.55
t 9.35
Ts 14.97
Ts 292
Ts 2.48
Ts + 3.54
DAY NUMBER
1s 1.00
Te 17.59
Ts 3.92
Ta §.78
ts 3.59
Ts 547
DAY NUMBER
Ts 8.66
Ts 1.56
Te 34.66
Ts $5.80
29.62
Ts 9.66
DAY NUMBER
Ts 4.09
Ts 941
Ts 5.36
Te 22.98
Te 10.79
Te 0.42

INTERNAL

EXPERIMENT NO=2

0

3.13 DEVIATION=
14.33 DEVIATIONS
4e45 DEVIATION@
19.97 DEVIATION=
26.77 DEVIATION=
26.97 DEVIATION=

1
SIGNIFICANT EME AN-CME AN =
SIGNIFICANT EMEAN-CME ANS
NOT SIGNIFICANT EMEAN-CME AI
SIGNIFICANT EME AN-CME Ai

6 NOT SIGNIFICANT EMEAN-CME AN=

SIGNIFICANT EME AN~CHME AN=
5
NOT SIGNIFICANT EMEAN-CME AN=
SIGNIFICANT EMEAN-CME AN
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT EMEAN~CME AN=
26
NGT SIGNIFICANT EME AN-CME ANS
SIGNIFICANT EME AN-CME AN =
SIGNIFICANT EME ANCHE AN =
NOT SIGNIFICANT EMEAN-CME AN=
NOT SIGNIFICANT EME AN-CME AN =
SIGNIFICANT EME AN-CME AN =
50

WOT SIGNIFICANT EME AN-CMEAN=
SIGWIF ICANT EMEA MEAN®
SIGNIFICANT EME AN-CMEAN®

SIGNIFICANT EME AN-CME AN=
SIGWIFICANT EME AN-CMEAN=
SIGNIFICANT EME AN~CME AN=
85

SIGNIFICANT EME An-CME AN &
SIGNIFICANT EME AN-CHE AN=

SIGNIFICANT
SIGuTF ICANT
SIGNIFICANT

EME AN-CMEAN®
EME AN-CME AN=
EME AN-CME AN®

SIGNIFICANT — EMEAN-CMEAN®
125

SIGNIFICANT — EMEAN-CMEAN®
SIGNIFICANT EME AN-CHE AN =
SIGNIFICANT — EMEAN-CHEAN=
SIGNIFICANT EME AN-CMEAN =
SIGNIFICANT EME AN-CHEAN®
SIGWIFICANT — EHEAN-CHEAN®

215 NUM
70 NUM

AUM=

*NUME

NUME
WUMS

DEG=
DEvs
DEG=
DEG=
DEG=
DEG=

DEG=
DEG=
DEG=
DEG=
OEG=
DEG=

OEG=
DEG=
VEG=
DEG=
DEG=
DEG=

DEGe
DEG=
DEG:
DEG=
DEG=
DEG=

DEG=
DEG=
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DEGe
DEGs

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DEG=
DEGs

eevee

PE RSSS SEEDes

PRESSE wevvwvw

ppepss

prpess

EMEANS
EMEAN®

EMEANS
EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN®

1.50
7.20
4645,
25.77
9.33
39.50

1.53
38.37
14.00
75.33
15.93
57.87

+90
45.00
14.70
33.05
24.40
47635

+90
85.50
20.27
69.33
10.97
38.47

1.40.
92.30
23.77
40.23
48.20
64.67

80
51.17
13.00
38.63
14.20
50.40

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEVs
EDEV=

EDEVe
EDEV=
EDEV=
EDEV=
EDEVs
EDEV=

EDEV=
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EDEV=
EDEV=
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EDEV=

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053
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200
242
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96

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CMEAN®
CMEAN=
CMEAN®
CHEANS
CMEAN=

CHEANS
CMEAN®
CMEAN®
CMEAN®
CMEAN®
CMEAN=

CHEAN®
CMEAN®
-CMEAN®
CHEAN®
CMEAN®
CMEAN®

CNEAN®
CHEAN®
CMEAN®
CHEAN®
CHEAN®
CMEAN®

CHEAN®
CHEANE
CMEANS
CMEAN®
CH EAI
CMEAN®

CHEAN®
CMEAN®

- CHEAN®

CMEAN®
CHEAN®
CHEAN®

25



SCREENED INTAKE
PUMP hw FOOT VALVE

i<-5.0cm 1D PVC

SAND FILTER
HOLDING TANK
(2x12x Lam)
+
I< 2.5cm PVC
CONTROL TANK > RETURN
@ (0.9 x11 9.0m) To eay
EXPERIMENTAL TANK
{0.9 x1.1x9.0m)
Z CHARCOAL, SAND
® INDICATES PVC BALL VALVES AND GRAVEL FILTER
TOP VIEW
+
E
CONTROL OUTFLOW—+09 3
(NO SLUDGE) ; To BAY L
eH
EXPERIMENTAL| OUTFLOW—+5
SEA WATER FROM | (+ SLUDGE) TO FILTER
HOLDING TANK
k 9.0m 4
e SIDE VIEW
SEA WATER—>|
SEWAGE SLUDGE —>|
SAND—>}

Diagram of experimental design, tank configuration,

Figure 2.
and sea water system.

22


‘TABLES

Number Page

1 Physical dimensions, number stocked and sample
size of animals, Experiment I and II....... eee e eee ee S

2 Taxonomic list of experimental organisms.............. 6

3 Results of trace metal analyses of NBS biological
standards, with minimum detectable concentrations...10

4 Initial trace metal concentrations of sewage sludge,
Experiment I and IL... ccc cece cece cee cee eee eee ee IS

ix


O€T

URCHIN

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
WICKLE
LEAD
ZINC

CAOK TUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROKIUM
COPPER
NICKLE
LEAD
LIKC

SHELL EXPERI:
DAY NUMBER 0
NO EXPERMENT MEAN= 2.93
NG EXPERMENT MEAN= 5.03
WO EXPERMENT HEAN® 4.45

NO EXPERMENT MEAN 14.30
Nu EXPERMENT MEAN] 31.83

NO EXPERMENT MEANS 4.37
DAY NUMBER 1
Ts 1,11 NOT SIGNIFICANT
Ts 1.06 NOT SIGNIFICANT
EMEAN®CHEAN
T= +62 NOT SIGNIFICANT
Ts 1.74 NOT SIGNIFICANT
Ts 1.94 NOT SIGNIFICANT
DAY NUMBER 5
Ts +44 NOT SIGNIFICANT
Te +21 NGT SIGNIFICANT
EMEAN=CHE AN
Te 1.37 NGT SIGNIFICANT
Ts 1.56 NOT SIGNIFICANT
Te 11.62 SIGNIFICANT
DAY NUMBER 25
Ts +41 NOT SIGNIFICANT
Ts 1.31 NOT SIGNIFICANT
EMEAN=CME AN
Te 1.75 NOT SIGNIFICANT
Te +67 NOT SIGNIFICANT
Te 3 NOT SIGNIFICANT

4
DAY NUMBER

Ts 225
Ts 73
Ts +55
Ts 210
Ts 1.17
DAY NUMBER
T= «30
Ts 2.78
Ts 4.44
Te +40
Ts 298
DAY NUMBER
Ts 1.58

T= 1.40
T= 1.68
Ts 1.23

Ts 2.60

50
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN=CNEAN
NGT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
65
NOT SIGNIFICANT
SIGNIFICANT
EME AN =CNE AN
SIGNIFICANT
WOT: SIGNIFICANT
WOT SIGNIFICANT
125
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN=CHEAN
NGT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT

MENT NG =2

DEVIATION=
OEVIATION=
OEVIATION=
DEVIATION=
DEVIATIGN=
DEVIATION=

EME AN-CME AN®
EME AN-CHE AN =

EME AN-CME AN®
EME AW-CME AN =
EME AN-CME AN®

EME AN-CMEAN®
EME AN-CMEAN®

EME AW-CME AN®
EME Ali~CME AN =
EMEAN-CMEAN =

EMEAN-CME AN =
EME AN-CME AN =

EMEAN-CMEAN®
EMEAN-CME AN®
EMEAN-CMEAN®

EME AN-CME AN =
EME AN-CME AN=

EMEAN-CMEAN®
EME AN-CME AN®
EMEAN-CME AN=

EMEAN-CMEAN=
EME AN-CMEAN®

EMEAN-CME AN@
EME AN-CME AN=
EME AN-CME AN®

EME AN-CMEAN=
EMEAN-CMEAN®

EMEAN-CMEAN=
EME AN~CMEAN®
EMEAN-CME AN =

015 NUM®
276 NUM®
200 NUN=
2.03 NUM=
1.11 NUNS
206 NUM®

013 OEGs
+23 DEGs

030 DEG=
13 DEG=
047 DEGs

013 DEG=
-207 DEG=

053 DEG=
1.56 DEG=
4.37 DEG=

+10 GEG=
237 OEGe

71.23 LEGS
1.53 DEG=
213 DEG=

-.03 DEue
237 DEG=

7237 DEGs
227 DEG=
1.63 DEG=

03 DEG=
71.30 DEG=

053 DEG=
7297 DEG:
2.43 DEG=

welT DEG=
1.00 DEG=

71.63 DEG=
2.97 DEG=
3.03 DEG=

SYEwOY

Pe PEE PH PEE FS

PR PEP PR EES

BEE

pane

EMEAN®
EMEAN®=
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEAN®
EMEAN=
EMEAN=
EMEAN®=
EMEAN®
EMEAN®

EMEAN=
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN=

EMEAN=
EMEANS
EMEAN®
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN®
EMEAN®
EMEAN=
EMEANS
EMEAN=
EMEAN=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEv=
EDEV=

EDEV=
EDEV=
EDEVs
EOEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

020
23

2.20
1.56

+10

79
279
1.36
1.41

168

CHEAN®
CMEAN®
CMEAN=
CHEAN=
CHEAN=
CHEAN®

CMEAN=
CMEAN=
CMEAN=
CHEAN=
CHEAN=
CMEAN=

CMEAN=
CHEAN=
CHEAN=
CMEAN=
CHEAN=
CHEAN=

CMEAN=
CMEAN=
CHEAN=
CMEAN=
CMEAN=
CHEAN=

CHEAN=
CMEAN=
CHEAN=
CHEAN=
CMEAN®
CMEAN=

CMEAN®
CHEAN=
CHEAN=
CHEAN=
CMEAN=
CMEAN=

40

2.77 CDEV=
5-23 CDEV=
4e4SCDE V=
14.83 COEV=
26.43 COEVe
4.60 CDEV=

2.67 CDEV=
5.07 CDEV=

29.93 CDEV=
15.60 COEV=

2.03 CDEV=
7.23 COEV=
4.45CDEV=

9.33 CDEV=

2.70 CDEV=
5.60 CDEV=
4.4SCDEV=
10.97 COEV=
39.57 CDOEV=
15.07 CDEV=

2.73 COEV=
9.43 CDOEV=
4.45CDEV=
14.53 CDEV=
44.00 CDEV=
10.37 CDOEV=

2.67 CDEV=
5.80 CDEV=
4.45CDEV=
16.70 CDEV=
37.03 CDEV=
11.40 CDEV=



CU THALASS!RA LCERVES—- EXP. 2

7}
= 2B 4
oz
a
S 4
~
Y coed
Ww
~
z 4
ti
Be.
St to
s
& & z
DAYS
cu URCHIN GUT- EXP. 2
Sy
=
2 ia 4
~
a
Ys 4
uw
4
Zz
£2 4
ui
=
-= T T T T T T T 7
s é B z
cCAYS

Figure 17. Mean-net uptake of Cu between experimental and control,
Thalassia leaves and urchin guts - Experiment II.

37


30

34

32

33

34

35

36

37

38

39

40

4d

42

43

Mean net uptake of Cr between experimental and
control, Thalassia roots and mangrove roots-
Experiment IT... 7. a)

Mean net uptake of Cu between experimental and
control, Thalassia roots - Experiment I............51

Mean net uptake of Cu between experimental and
control, Thalassia roots and mangrove roots-
Experiment ID... ccc cece eee eee eee eee ne OS

Mean net uptake of Pb between experimental and
control, Thalassia roots - Experiment I............53

Mean net uptake of Pb between experimental and
control, Thalassia roots and mangrove roots-
Experiment IT... cece ccc cee eee ce ce teen ee OF

Mean net uptake of Zn between experimental and
control, mangrove roots - Experiment I..... wee ee OS

Mean net uptake of Zn between experimental and
control, Thalassia roots and mangrove roots-

Experiment IT. eee cece eee eee eee see eee 56
Mean net uptake of Cd between experimental and

control, Thalassia roots - Experiment II...........57
Mean net uptake of Cr and Ni between experimental

and control, sea cucumber guts - Experiment I......58
Mean net uptake of Cd and Cr between experimental

and control, sea cucumber guts - Experiment II.....59
Mean net uptake of Cu between experimental and

control, sea cucumber guts - Experiment I...........60

Mean net uptake of Cu and Ni between experimental and
control, sea cucumber guts - Experiment II.........61

Mean net uptake of Pb and Zn between experimental and
control, sea cucumber guts - Experiment I...........62

Mean net uptake of Pb and Zn between experimental and
control, sea urchin guts - Experiment II............63

viii


ZN FOULING ORGANISMS— EXP. 1

'ea

4

122

MERN ¢E-C) / PPM

N
8
pisiit

DAYS

ZN SEWAGE—- EXP. i)

asaa
amas 4
zsme2 4
zone 4

1sam J

MEAN PPM

imme 4

a4
@h
wa

Bz

Figure 11. Mean net uptake of Zn between experimental and control,
fouling organisms and sewage - Experiment I.

31


TIT

EXPERIMENT NO=1

SIGNIFICANT
SIGNIFICANT
SEGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

STONLFI CANT
SIGNIFICANT
SIGNIFICANT
SIGNIFI CART

SIGiHIF ICANT

2DO PLANK SOLIDS
DAY NUMBER 25
CADMIUM +00 NGT
CHRGMIUM +00 WGT
COPPER +00 WOT
NICKLE T= +06 NCT
LEAD +00 WOT
ZINC Ts +00 wCT
OAY NUMBER 59
CACH [Us +00 aGT
CHRGE UM 1.34 NOT
COPPER 1.37 NOT
NICKLE = T= 1.29 NUT
LEAD 32942.04
ZINC +00 wCT

SIGNIFECAKT

EME AN-CMEAN =
ENC An~CME an =
EME An-CHi ON
tT AN-CME ON
EME AN-CME AN =
EME AN-CME ANS

EME An-CME ANE
EME An-Ché dns
EME AN-CME ANS
EWE Aw-Citt Awe
EME ACME An =
EME AN=-CME ANS

-.80
~15 240
-15.10
46.30

40
59.70

-.60
7153.90
~40.95
-115.20
739.40
-16.40

DEGE
ObGs
GEGs
vEGs
DEG:
LEG=

DEGe
LEG
DE
DEU=
DEGs
DEG

Oreo eoocce

EMEANS
EMEANS
EMEan®
EMEANS
CHEANS
EREANS

EMEANS
EMEAN®
EMEAN=
chEAN=
EMEAI
EME AWS

4.90
185.40
74.60
201.60
15.70
126.70

2.30
10.00
19.20
49.50

7.80
30.40

EDEV=
EDEV=
EDEVe

EDEV=
EDEV=
EOEV=
ELEV=
EOtv=
EDEV=

+00 CMEAN=
00 CMEAN=
200 CHEAN=
200 CMEAN=
200 CHEAN=
200 CMEAN=

200
00
+00
00
200
+00

CMEAN=

CHEAN=
CMEAN=

2k

5.70 COE
260.80 CDEV=
69.70 CDEV=
248.10 COEVs
15-30 CDEV=
67.00 CDEV=

2.90
163.90
60.15

CDEV=
CDEV=
CDEV=
COEV=
COEV=
CDEV=

47.20
46.80

+00
+00
200
+00
+00
60

+00
93.90
24.40
72.97
+00
200


The ground sample, not used for analysis, was stored in plastic,
screw cap bottles. The dried samples were placed in cleaned

Pyrex beakers, at least lg where possible, and were digested using
the following procedure.

Twenty-five m2 of a concentrated HNO,: HC& mixture (3:1)
was added to the dried, weighed sample in the covered clean beaker.
The sample was slowly boiled and refluxed in the covered beaker
at 90 to 95°C until 1 m& of solution remained. The sample was
allowed to cool then 30 m& of 30% H,0 was added slowly and re-
heated to 90-95°C until 1 m& of the sdlution remained. A further
10 m2 of HNO,:HC2 mixture was added and reheated. The mixture was
allowed to e801 and 10 mg of 30% H,0, was added. This wa: allowed
to boil again (90-95°C). The 10 m& 4dditions were repeated, if
necessary, a maximum of three times, Six m& of 2N HC& were added,
boiled at 90-95°C, until 3 m& were left. The sample was allowed
to cool then centrifuged in a clean 50 m& plastic capped centrifuged
tube at 1700 rpm for 20 minutes. The liquid was carefully decanted
and brought to 10.0 m& with R.O./D.I. water. This sample was
placed in clean, 25 m& polyethylene vials with caps.

ll


The organisms, except for the clams, were all collected
using manual techniques at Pta. Ostiones, Puerto Rico. The
organisms were allowed to equilibrate for 2 weeks in the flowing
sea water system. The turtle grass was replanted manually at
400 plants m “, after soaking in a 10% NAPDH solution (Kelly
et al., 1971). The clams were collected at Pta. Viento on the
southeast coast of Puerto Rico.

Mangroves in Experiment I (230 seedlings) were planted as
ungerminated seedlings and as germinated seedlings in Experiment I
(120 seedlings).

Oysters in Experiment I, were placed in tanks and suspended
in one plastic mesh bag per tank. In Experiment II the oysters
were placed 13 per bag, for a total of 10 bags per tank. The
location of the bags was randomized except no bag was hung directly
over the sewage sludge deposit.

Table 2 is a species, common name list of plants and animals
used for this study.

TABLE 2
Common name Species
Clam Codokia orbicularis
Oyster Crassostrea rhizophora
Snail Nerita tessplata
Sea cucumber Holothurea sp. (predominently
— H. mexicana)
Urchin Lytechinus variegatus
Turtle grass Thalassia testudinum
Red mangrove Rhizophora mangle

After equilibration, organisms were sampled at the pre-
scribed intervals, the samples were removed, physical measure-
ments made, placed in double bags and frozen. Simultaneously,
samples were collected for hydrocarbon analyses. The samples
for hydrocarbon analyses were placed in glass bottles with
aluminum foil seals, capped and frozen. The bottles were previ-
ously rinsed with 2N HC2, deionized water, CC&,, and air dried.
The physical dimensions for the animals sampled and number of
animals sampled per day are shown in Table 1.

Overlying Water

Water samples were taken, for Experiment I and II, prior
to biological samples. There were three types of water samples

6


CONTENTS

Foreword. ......e.ee Fn ef
ADSTPACTL Cee cece eee eee eee tee eens ee eee ween e nes eeeee GUL
FIg“uresS.. cc. cece cece eee eee ee eee scenes beeen eee wv
Tables........ rare 2 is a
Acknowledgment... cc eee eee cece eee ee eee eee eee eee ener ee eeee eee e ix

2. Imtroduction.. cece ccc cece cee cee tence teen ne neee

2. Methods... cece cc ccc cece cece cece cent e ee eee teen tees

Construction of sea water tanks.....

Sewage sludge............. beeen eee eee ee ee ee eens

Sampling.........0+0- beeen eee ee ee ete eens
OPZANISMS.. eee cee eee eee eee eee ete eens
overlying water... .. cece eee ee ee eee scene eee
POPE WATER... eee ccc c cece ee neee see see eens
sediment............0. see e ee eerece ees
dissolved and particulate trace metals..........

Physical and chemical analyses............ wrens
PENEVAL. cee ee ce ccc cece eee eee
oxygen, temperature and salinity..
dissolved reactive phosphate.....
particulate trace metals

soluble trace metals..... sees ete rene oe

biological and sediment samples.......sc..e sees

3. RESULTS. . ere eee eec cere eee eee eee eee neeee

4. Discussion............... beeen een ete e eee

§. Conclusions and Recommendations.............- bce wees
Bibliography... cece cece eee eee ee eee eee eee ee eee e ee eee teen ees
Appendices

A. Sorted raw data, Experiment I and II................ see

B. Results for net uptake for Experiment I and II..........

C. Experiment II, water data..... eee eee eens sae e eee
D. Wet/dry ratios for all samples............ eden eee e eee


€eT

EXPERIMENT NO #2

3.07 DEVIATION=
7247 DEVIATION=
4.45 DEVIATIONS
18.27 DEVIATIGN=
36.17 DEVIATIONS
3.60 DEVIATIONS

NOT SIGNIFICANT EMEAN-CHEAN®

QYSTER SHELL
DAY NUMBER 0
CAON1UH NO _EXPERMENT MEAN®
CHROMIUK NO EXPERMENT HEAN®
COPPER =~ NO EXPERMENT MEAN=
NICKLE = -NQ EXPERMENT MEAN
LEAD NO EXPERHENT MEANS
LINC NU EXPERMENT NEAN®
DAY NUMBER 1
CADMIUM T= +29
CHROMIUM T= 3.25 SIGNIFICANT
COPPER «T= 1.00 NOT SIGNIFICANT
NICKLE = Te 10.58 SIGNIFICANT
LEAD Te +78 NOT SIGNIFICANT
ZINC Ts 6676 SIGNIFICANT
DAY NUMBER = 5
CADMIUM T= +94 NOT SIGNIFICANT
CHROMIUM T= +66 NOT SIGNIFICANT
COPPER =T# 1.51 NOT SIGNIFICANT
NICKLE Te 1.19 NOT SIGNIFICANT
LEAD Ts +74 NCT SIGNIFICAKT
ZINC Te 2.02 NOT SIGNIFICANT
DAY NUMBER 25
CADMIUH Ts 3446 SIGNIFICANT
CHROMIUM T= 4.07 SIGNIFICANT
COPPER «T= 2.00 NOT SIGNIFICANT
NICKLE = Te 2.22 NOT SIGNIFICANT
LEAD Ts +39 NOT SIGNIFICANT
ZINC Ts 3.61 SIGNIFICANT
DAY NUMBER 50
CADMIUM Te +25 NOT SIGNIFICANT
CHROKIUA T= 1.00 NOT SIGNIFICANT
COPPER EXEAN SCRE AN
NICKLE = T# +42 NOT SIGNIFICANT
LEAD Ts +62 WOT SIGNIFICANT
ZINC Ts 268 NOT SIGNIFICANT
DAY NUMBER 65
CADMIUA T= 1.89 NOT SIGNIFICANT
CHROMIUM T= 1.34 NOT SIGNIFICANT
COPPER T= 5.24 SIGNIFICANT
NICKLE = T= +23 NOT SIGNIFICANT
LEAD Te +77 NOT SIGNIFICANT
ZINC Ts 2.61 NGT SIGNIFICANT
DAY NUMBER 125
CADMIUH T= «71 HOT SIGNIFICANT
CHROMIUM Ts 4a1k  SIGWIFICANT
COPPER = T= 1437 NOT SIGNIFICANT
NICKLE | T# 5.72 SIGNIFICANT
LEAD 1 +30 NOT SIGNIFICANT
ZINC Te 13.46 SIGNIFICANT

EME AN=CME AN®
EMEAN-CMEAN®
EME AN-CME AN =
EME AN@CME AN®
EME AN-CMEAN®

EME ANCHE AN®
EME AN-CHE AN =
ENE AN@CME AN®
EME AN-CHE AN®
EME AN-CMEAN®
EME AN-CMEAN®

EMEAN-CMEAN®
EME AN-CME GN ®
EME AN~CME uN &
EME AN-CMEAN=
EME Aw-CME AN®
EME AN-CME ANS

EME AN-CME AN =
EME AN-CME Gh =

EME Aiv-CME AN®
EME AN-CME BN ®
EME AN-CHE BN®

EME ANCHE ANS
EME AN-CME AN ®
EME AN-CME AN®
EME Au-CHE AN =
EMEAN-CME AN=
EME AN=CME AN®

EME AN-CHE AN®
EMEAN-CME AN®
EME AN-CHE AN®
EME AN-CME ANS
EME AW-CME AN =
EME AN-CME AN®

+06 NUH®
215 NUM®
+00 NUNS
242 NUM®
645 NUNS
046 KUMe

07
72630
1.58
4.07
7663
~6.03

733
223
~.27
71.60

Swuwow

DEGs
OEGs
DEGs
OEG=
DEGs
DEG=

DEG=
DEGs
DEG=
DEG
DEG=
DEG=

DEG=
DEG=
DEG
vEGe=
DEG=
DEG=

Eos
vEGs

DEGs
DEG=
DEG

DEG=
LEG=
DEG
vEGe
DEGs
DEG=

DEG=
DEG=
LEG=
DEG=
DEG=
DEG=

PEER EH PSS EL PEE es

ne

Ree

PREESS PEEEEE

EMEAN®
EMEAN®
EMEANS
EMEAN®
EMEANS
EMEANS

EMEAN®
EMEAN®
EMEAN®
EMEANS
EMEAN®
EMEAN®

EMEAN
EMEAN®
EMEAN®
EMEANS
EREAN®
EMEAN=

EMEANS
EMEAN®
EMEAN®
EMEAN®
EMEAN®
EMEAN®

EMEANS
eMEANS
EMEAKs
EMEANS
EMEANS
EMEAN®

EMEAN®
EMEANS
EMEANS
EMEAN®
EMEAN®
EREAN=

CKEAN®
CHEAN=
CMEAN®
CHEAN®
CMHEANS
CHEAN®

CHEAN®
CHEANS
CHEAN=

8 CHEAN=

CMEAN®
CHEAN®

CMEAN=
CMEANS
CHEAN=
CHEAN=
CMEAN=
CAEANS

CHEAN®
CHEAN=
CMEAN®
CHEANS
CMEAN®
CHEAN>

CHEAN®
CHEAN=
CHEAN®
CMEAN=
CHEAN®=
CMEAN®

CHEAN®
CMEAN®
CHEAN®
CMEAN®
CHEAN®
CHEAN®

3

COEV=

O CDEV=

18.07
35.37
6.70

1.80
9.70

COEV=
CDEV=
CDEV=
COEV=

COEV=
CDEV=
COEVe
COEV=
COEV=
COEV=

COEV=
COEV=
CDEV=
CDEV®
COEV=
CDEV=

COEV=
COEV=

G.G5CDEV=

13.37
46.57
6.93

1.80
11.97
10.93
14.50
50.93
15.70

2.37
21.30
9.77
21.10
39.20
10.67

CDEV=
CDEV=
COEVe

COEVe
COEV=
COEV=

COEV=



TOT

EME AN-CME AN =
EME AW-CilE AN =
EME An-CME AN ®
EME AN-CME AN=
EME AN-CME AN =
EME AN-CME AN =

DEVIATION=
DEVIATION
DEVIATIUN
DEVIATION=
DEVIAT LuN=
DEVIATIUN=

NOT SIGNIFICANT EME AN-CMEAN=

EME AN-CME AN =
EME AN-CME AN=
EME AN-CME AN=
EME AN-CME AN=

EME AK~CME AN =
EME AN-CHE AN=

EME AN-CME AN=
EME AN-CME AN

EME AN-CME AN=
EME AN~CME AN=
EME ACME AN=
EME AN-CME ANE
EME AN-CME AN=
EME AN-CME AN =

OEVIATION=
DEVIATICN=

9.99 DEVIATION=

NERITA INTERNAL EXPERIMENT NO=2
DAY NUMBER 0
CADKIUM T= 200 NOT SIGNIFICANT
CHROMIUM T= 200 NUT SIGNIFICANT
COPPER Te +00 NOT SIGNIFICANT-
NICKLE Te +00 NOT SIGNIFICANT
LEAD Ts: -00 NOT SIGNIFICANT
ZINC Te +00 NOT SIGNIFICANT
DAY NUMBER 1
CADMIUM NO EXPERMENT MEAN= +15
CHROMIUM NO EXPERHENT MEAN= 1.70
COPPER  =ND EXPERMENT MEAN= = 62.25
NICKLE = -NO EXPERMENT KEAN 45
Lead NG EXPERMENT HEAN= 23.30
ZINC NO EXPERMENT HEAN= 173.60
OAY NUMBER 5S
CADMIUM Ts +00
CHROMIUM EMEAN=CHEAN
COPPER «T= +00 WUT SIGNIFICANT
WICKLE = T# +00 NOT SIGNIFICANT
LEAD iid 200 NCT SIGNIFICANT
ZINC Ts +60 NOT SIGNIFICANT
DAY NUMBER 25
CADH 1M EMEAN=CKEAN
CHROMIUM T= +00 NOT SIGNIFICANT
COPPER «T= 200 NUT SIGNIFICANT
NICKLE EMEAN=CHE Aly
LEAD TM +00 NOT SIGNIFICANT
ZINC 1 200 NGT SIGNIFICANT
DAY NUMBER 50
CADMIUM T= +00 NOT SIGNIFICANT
CHROKIUM T= 200 WGT SIGNIFICANT
COPPER «f= +00 WCT SIGNIFICANT
NICKLE = Te 200 NOT SIGNIFICAKT
LEAD Ts +00 NGT SIGNIFICANT
ZINC Ts 200 NOT SIGNIFICANT
DAY NUMBER 125
CADNIUK NG EXPERMENT MEAN 3.00 DEVIATIUAS
CHROMIUK NO EXPERMENT HEAN® = 41.60
COPPER NO EXPERMENT NEAN= 21.40
NICKLE «=—-NG EXPERMENT MEAN=
LEAD NO EXPERMENT MEAN® 3689

ZINC

NO

DEVIATIUN=

EXPERMENT MEAN® 62.30 DEVIATION]

1.90 DEG=
7.30 DEG=
78.30 DEG=
-+40 DEG=
20.10 DEG=
9.60 DEG=

+00 NUNS
+00 NUM=
290 MUMS
00 NUM=
200 NUN
+00 NUM=

2.95 DEG=

8.10 DEL=
23.55 vEvs
“10.60 DEG=
711.60 DEG=

5.70 DEG=
6.40 DEG=

220 DEG
770.60 VEG

+90 DEG=
3.90 DEG=
3-10 DEG=
=.90 DEG=
10.50 DEG=
26.70 DEG=

+00 NUM= 1
200 NUM= 1
+00 MUd= 1
+00 HUM= 1
+00 NUM= 1
+00 NUM= 1

ecco0e

©@00000 co oo

EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAW=
EMEANS
EMEAN=
ENEAN=
ENEAN=

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEANS

5.40
10.80
46.90
27.00
40-90
119.70

3.10
1.70
58.40
24.00
18.10
157.00

1S
1.70
56.00
45
34.20
116.50

4.10
5.60
53.80
16.40
6020
168.50

EDEV=
£DEV=
EDEVe
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=

00
00
00

00

+00

CMHEAN=
CMEANS
CMEANS
CMEAN=
CHEAN=
CHEAN=

(MEANS
ChEAN=
CMEAN=
CMEAN=
CMEAN=
CMEAN=

CMEANS
CMEAN=
CHEAN®
CMEAN=
CMEAN=
CHEANS

GMEAN=
CMEAN=
CMEAN®
CMEAN=
CMEAN=
CMEANS

10

3.50 CDEV=
3.50 COEV=
55.20 CDEVe
27.40 CDEVE
20.80 CDEV=
129.30 CDEV=

15 COEV=
1. 70COEV=
50.30 CDEV=
045 CObVe
28.70 COEVe
168.60 COEVe

el SCOEVE
7.40 COEV=
49.60 CDEV=
*G5CDEV=
34.40 CDEV=
189.10 CDEV=

3.20 COkVs
1.70 CDEV=
50.70 CDEV=
17.30 COEVE
16.70 CDEV=
141.80 CDEV=

+00
+00
+00
009
00



09

Figure 40.

CU HOLOTTHURIAN GUT- EXP. |

3@ 4
=
a 4
ou
~“
ze J
~
Vv
1 4
lJ
- wed
Zz
fund 4
lJ
= ia} 4
T T T T T ms T 1
=} <= o -
i=] i] nN
i=}
DAYS

Mean net uptake of Cu between experimental and control, sea cucumber guts - Experiment I.


SECTION 4
DISCUSSION

Oysters and clams have been extensively studied regarding
trace metal uptake because of their commercial significance,
hardiness and feeding method. Bivalves, in general, show uptake
and concentration of most trace metals (Kerfoot, W.B., 1973;
Kerfoot, W.B. and S.A. Jacobs, 1973 a and b; Shuster, C.N. and
B.H. Pringle, 1969; Huggett, R.J. et al., 1973; and Frazier, J.N.,
1976). However, oysters and clams in our research showed no
consistent nor significant net uptake of metal except possibly
zinc and copper. In Experiment I, all the oysters were placed
in a single net bag. This procedure was changed to ten net bags
per tank in Experiment II. This drastically reduced the crowding
and increased the survival of the oysters beyond day 50 in
Experiment II. The wet weight of the clam and oyster internal
organs was too small to allow an adequate sample size for the
trace metal analyses. This same problem was encountered in the
analyses for the snails, holothurian muscle parts in Experiment
I, and zooplankton samples for both Experiment I and II.

The results for lead in the sewage sludge were very low
compared to other investigators (Table 4) and are due to the
addition of H,S0 to aid in the digestion of the sludge organic
matter. When*the sample was heated, the lead was lost as volatile
PbSO,. The H,S0, was only used for the digestion of the sludge.

The results for the analyses on the NBS bovine liver standards
(Table 3) are in good agreement for Cu and Zn and high for Cd.
Our results for Cr and Pb in bovine liver were below our de-
tection limits. The results for NBS orchard leaves were in good
agreement for Cu, Pb and Zn. Cadmium, in NBS orchard leaves,
was too high as was nickel. The coefficient of variation (CV)
for Ni was lower than the CV for Cd. The results for Cr, in
orchard leaves, were in good agreement with the non-certified
value for Cr published by NBS.

Our results for the concentration of trace metals in water
for Experiment I were at or below the detection limits for
nearly all metals and were therefore not shown. This was due
to the 2.5 & sample size. When we used 4 2 sample in Experi-
ment II, the results for most of the metals were within our
detection limits. However, no significant difference was noted
between metal concentrations in the experimental tank and those


Mean Wet/Dry Ratio (x) with n (number of samples) and Standard
deviation(SD) for all organisms sampled in Experiment I and ITI

Clam
Gut
Shell
Sea
Cucumber

Body
Gut
Muscle

Urchin
Gut
Shell

Snail
Gut
Shell

Oysters
Gut
Shell

Thalassia

Leaves
root/
Rhizomes

"Fouling
organisms"

Sewage
Sediment

Mangroves

Bottom of
hypocotyl

Meristem
above
hypocotyl

Leaves

Roots

12

10
10

oo

10

+

BRE
oo

Ho
Ro

10.2
7.5

5.2
14.8
1.4

S.D.

1.80
0.02

£70
190

oR
oR

0.60
41.1
1.0

134

N x S.D.
13 10.7 1.20
12 1.4 0.00
12 4.5 0.40
13 10.6 3.20
Not determined, combined

with gut
12 10.7 2.30
13 2.0 0.20

7 3.9 1.0

7 1.41 0.0
12 11.6 2.10
13 1.3 0.20
12 12.8 2.90
13 9.9 2.2

7 5.0 1.4

7 8.7 ail
13 1.4 0.10

7 3.4 0.60

7 4.0 0.40

6 . 1.3

7 §.6 0.50


Eisler, R., 1971; Gardner, G.R. and G. LaRoche, 1973; Jackim,

E., 1973; Frazier, J.M., 1976; Ferrell, R.E. et al., 1973;
Huggett, R.J., 1973; Hannan, P.J. and C. Patouillet, 1972).
Schroeder (1975) demonstrated, using radioactive isotopes, that
Thalassia testudinum would concentrate cobalt and manganese in

the leaves rather than the root structure (excluding rhizomes).
The incorporation of cations in Thalassia testudinum was primarily
accomplished through the leaves (Schroeder, 1975).

The use of sewage sludge to amend agricultural soils has
been extensively studied (Street, J.J. et al., 1977; Silviera,
D.J. and L.E. Sommers, 1977; Turner, R.E. et al., 1976). However,
very little has been published regarding the rates of release
of potentially toxic trace metals into complex tropical eco-
systems. The necessity for this type of controlled field experi-
ments was explained by Menzel (1977) for the CEPEX experiment.
Inherently, it is impossible to duplicate exactly the marine
ecosystem due to its spatial and temporal dimensions. Also,
the isolation of organisms from their surrounding environment
produces results that are ambiguous. Therefore, it is unlikely
that any "artificial" system can duplicate conditions in the
natural environment.

It may not be necessary-to duplicate in a laboratory the
real system, but rather to produce a system that can duplicate
itself (Menzel, D.W., 1977), while at the same time attempting
to duplicate the gross physical marine parameters, such as light,
suspended sediment load, salinity and temperature. In order to
do this, a flow-through sea water system with replicate experi-
ments in different seasons is mandatory.

Our objectives in this research were to use a flow-through
system of sufficient size and complexity as to nearly duplicate
a tropical marine ecosystem and also allow repeatability over
varying seasons. This system would allow us to determine the
rates of uptake, by a tropical marine community, of Cu, Cd, Cr,
Zn, Ni, and Pb which were leached from sewage sludge by sea
water. This community was comprised of turtle grass (Thalassia
testudinum) with its associated sedimental infauna, spiny sea
urchin (Lytechinus variegatus), sea cucumber (Holothurea mexicana) ,
attached "fouling organisms", plankton and the red mangrove

(Rhizophora mangle).

This ecosystem was chosen for three reasons: (1) the system
comprised a tightly bound community with strong interaction
between the members, (2) the important role of the seagrass/
mangrove ecosystem in the overall tropical near-shore environment
(Jones, J.A., 1968; Odum, H.T. et al., 1959; Golley, F. et al.,
1962), (3) the possibility of the active concentration of these
trace metals in the higher trophic levels with passage of the
materials through the food webs.



SD soc MEAN SIC SD SIC

MEAN SOC

MEAN PART SD PART

METAL

E/C

nO

oo

oo
oo

+00
04

oo
oo

da

oo

dd
oo

Cd

no
oo

on

Ceres
oo

oO +
oo

dd
oO +

om
oO +

Cr

-50

ao

acd

a+
oo

ww

oo

No
aN

0 =
aN

Cu

AN

od

on
wot

Oa
oo

tn
oo

ao
oo

+o
tot

Ni

on
aa

mw
ON

oo
oo

eo
oo

N/A
N/A

oo

Pb

ost
oo

0.78
1.8

oo
dN

Zn

132


The authors would not recommend, based on the results
of this research, the dumping of sewage sludge in shallow,
tropical marine environments. The potential exists for rapid
concentration of toxic trace metals by members of the food web
high on the trophic scale. These toxic levels could then affect
organisms used as food by man.

The next logical step in future research would be to add
vertebrate organisms to this system to see if a herbivore, which
normally consumes turtle grass, would show a net uptake of toxic
trace metals.

21


eccceccccecccececececcececeecacecececcacececececccececaccccecca

PADDY DDG WWW WDD WU ee ee et ree tt tt tnt et a edt a pet et tt Ot pet nt tet tt tod tet tt re tt te
RYUNRNNNNNNNNNNNNEYNNNNNUN NANNY NNN NNN NN E NR NNN RN NUNN

COPPER OOOH RE HOO OHH EE RH OOO HH OOOH HER OOORP HEH OOOKF RH OOOH HOO

SQRTED DATA 20
1 1.9 8.9 7.5 20.0 21.8 22.0
1 2.0 9.0 7.3 20.2 20.2 27.2
1 2.0 8.2 764 19.6 18.7 26.8
1 2.0 8.0 7.2 16.3 17.4 2661
1 2.1 7.8 Tel 18.7 16.9 27.5
5 1.8 17.3 10.9 40.5 18.4 39.5
5 1.7 18.9 11.6 38.8 17.9 44.9
5 1.8 19.9 11.5 38.2 18.2 45.2
5 2.1 14.8 9.1 30.9 13.5 34.0
5 2.1 14.9 9.4 32.0 15.0 36.2
5 2.2 15.0 904 32.4 11.6 35.7
25 1.4 35.0 14.5 57.5 25.4 41.3
25 1.5 39.4 14.7 54.8 26.9 39.4
25 1.4 40.6 16.0 64.2 26.8 43.5
25 1.7 22.5 10.7 28.9 22.5 30.3
25 1.6 15.4 11.3 29.6 2361 31.7
25 1.7 14.3 11.6 29.5 20.3 31.6
50 262 13.9 11.5 17.1 24.5 24.5
50 262 13.3 10.7 15.6 23.4 30.7
50 202 14.5 10.8 16.0 22.5 26.9
50 2.1 15.8 11.8 19.4 23.7 35.1
50 202 15.4 11.6 20.2 23.0 32.1
50 262 14.4 11.7 18.7 24.3 28.1
85 2.3 71.2 15.6 48.6 19.3 42.5
85 2.4 70.3 15.6 46.9 20.9 42.6
85 204 70.8 15.2 47.0 20.7 42.5
65 202 27.0 9.5 20.4 13.7 41.0
85 2.2 28.8 11.0 21.2 14.7 42.2
85 262 29.5 11.0 23.2 15.8 42.2
125 2.3 2626 20.4 31.6 44.0 50.2
125 264 29.6 21.1 36.7 49.7 56.3
125 2.4 28.6 20.9 35.3 47.0 57.6
125 21 17.6 9.7 31.6 20.7 39.4
125 2.1 17.7 9.3 29.8 24.8 33.4
125 1.9 17.6 10.5 30.7 23.8 35.3
0 2.8 562 6.0 12.5 30.8 4.3
0 2.9 5.7 6.0 13.9 31.7 4.4
0) 3.1 4.2 4.8 16.5 33.0 404
1 3.1 542 543 13.9 29.7 5.2
1 2.9 526 51 15.4 31.7 5.0
1 2.7 56 5.8 14.3 30.3 5.0
1 2.8 5.5 5.8 15.2 30. 5.0
1 2.7 503 526 14.6 28.2 46
1 2.8 4.9 51 14.7 26.7 4.2
5 204 5.3 5.9 13.4 26.8 19.4
5 3.3 4.8 58 14.0 28.9 20.0
5 2.7 409 6.0 12.7 29.6 20.5
5 2.7 5.4 5.6 14.0 30.8 15.4
5 264 4.5 5.0 14.0 29.8 15.4
5 2.9 5.3 4.9 13.7 29.2 160.0
25 1.9 7.3 5.1 8.1 4726 9.0
ra 2.6 7.5 5.3 9-3 48.0 8.8

89


APPENDIX C

Experiment II, Water Data

The column headings are defined as follows:

1. E/C

2. METAL

3. MEAN PART

4, SD PART

5. MEAN SOC

6. SD soc

7. MEAN SIC

8. SD SIC

E is data from experimental tank
coum " control "

concentration (ug/k) shown are for the defined
metal, either Cd, Cr, Cu, Ni, Pb or Zn

values shown represent mean (yug/2) concentration
for particulate fraction

represents the standard deviation of the mean
particulate function

values shown represent mean concentration (yg/2)
for the soluble organic fraction (Retained on
Amberlite resin)

represents the standard deviation of the mean
soluble organic fraction

values shown represent the mean concentration
(yg/%) of the soluble ingrganic fraction
(Retained on Chelex-100 resin)

Represents the standard deviation of the soluble
inorganic fraction

131


13

SORTED DATA

r-) MMOIARVOVANTANNOR AE OR ATNOTON AR ADNNAON OR NAITONRENNQIAM
eee oo estes ot eee ere ena eee seeeeeo eee eee ee oe e ee ee ee ee eee
BAAR ONNNSE MTA ENS usw ANETTA TN TTR DODOOLTI MNS NE OEM AROOEFLTOON
ratas ade Ata

9.2

0
4.9
3.4
4.0
7.9
4.5

AAAHRAARANKROKRHHK oa
cose eee ee eo oe ee eee eee we Oe wee owe ooo eee Oe eee eee .
MOOR END AHASFURODOAARAHARABAAHRAARANAMOBRHARARAAABRHARH BA oa
anny RAD a DAARDRAKRARAAH a AARAARARAH BD AAK oa
DOD oe DHARFATABRARAT oa DARADAAAH BRB BHRG on
tre ' riettedtotrrerers f) peeve eer tet ot toe '
DOROVUHADARTAOMMOMN ON SOM OMAN QOMAMOCCOONNERNNHOANHOVAAARQOAM
coe cee eee ee cc eeeceveceeeer eee eee eceec ere eee er eeeeeere eee ee
TN ARHRRORO KKH AM AMATATONSTS IAG ONMNANANMNTHETEMINNNNEO
am Ne - aa
WOM DALAM ONVANAAAMOTT TOON AREMAKROMOONATONATIOONNER NAY
eee we ew eee ewes e rere s reese ee eres eeeeeeeeeeeeeeces eee
AMAORFOTFANDAMMTATERADAMOT ET FANON OM SCOUR RE AOL ES OOHMMANG
at ot awe
AONE TROANTEOFTOOALNHNHOAMOHNNNHROTDANMODOOAIHOOMAONE MAAN
Pee er ee er een en re rere a ee ee ee De re)
SNBTHOSSSMSOMHATTS ES OME SSSNANASEOSSNAN AAA AEM ADAM ANA AA
mA aa
DONOOMANM TT TOLER AANNANMHAMEMMM FINN ODOR OFAN AANE TITONST TOE OD
er ar ee i er a a ee a  D
aaa ae
MNNNNOCONNNANNNOOCCOOONNNHNNHNOCONNNNNODDODOOONNANNNNOOCONNN
NNN WNW AWIANANNN NA DANNANANA NAN
atte Adtdtdde|” Adadae

DOA MAA AHODCO AD ADOCOHANHOOCO HAMA MAO O MH MAHODOO HN AHOOCO NAMM AMAMOOO
NBN NNNNN NNN NN NNN NNN NN NNN NNN NNN NNN NNN
A DANN DMODNANDMNAAGBMDDAMAMMIGVVOOVYOVYVOVYVYYYVOYVYUYVUSII Is
BESTEL ETA SESEAAATEATAALAAATATEETETATLATLATEEREAATTEATLEAEATIEE

82


The results from this research will aid in forming a basis
for determining the potential effects of leached toxicants from
sewage sludge on a tropical marine ecosystem. These problems are
especially critical in Puerto Rico. The population density on
the island is nearly 2000 people per square kilometer; about
ten times that of the continental United States. Only a few
towns and cities have sewage treatment facilities and most of
the sewage is dumped, minimally treated, into the rivers and bays.

With the increased awareness of the potential detrimental
effects of raw sewage on the health of the inhabitants and the
marine ecosystem, the construction of sewage plants will increase.
This increase of modern plants will produce large amounts of
sewage sludge. The disposal of this sludge will pose an especially
critical problem in the densely populated, mountainous island of
Puerto Rico as the available land for sludge disposal is very
limited and disposal poses its own set of ecological problems
at least as serious as marine disposal of sludge. Therefore, the
judicious selection of ocean dump sites for sludge, if this
alternative is chosen, will require estimates on the leaching
rates of toxic substances from sewage sludge by sea water. These
substances can enter the food web and become concentrated relative
to the concentration in the water column. The toxicity to marine
organisms of Cd, Pb, Cr, Ni, Zn and Cu has been demonstrated
(Eisler, R., 1971; Eisler, R. et al., 1972; Frazier, J.M., 1976;
Gardner, G. and LaRoche 1973; Ikuta, K., 1968). It is also
possible that some of the leachable substances could promote
growth in portions of the ecosystem, i.e., vitamins, chelators,
organic growth substances (Vallentyne, J.R., 1957). However, the
potential possible benefits of sewage sludge leachate will not
be examined in this research.


CCCCHAH AANA AAA A tH ttt ttt
He WDD DDD DODO DDD DDD DDD DDD DD DD DDD DNDDDDDBOE CECE Oe
NNN NNN RNR DNDN DD RR ROR RO Pt
OPP ERE HOO OPH HOO OH HH OO OR HRM ODOR EH OO OP RH OOO HM em Ee HOR mE HOO

SORTED OATA 19

95 3.6 128.4 2961 73.4 6.8 31.6
85 3.4 129.3 29.1 74.0 607 32.5
85 2.5 43.1 22.4 29.3 6.3 20.6
65 2.7 49.5 23.8 32.0 603 27.0
85 2.7 42.4 24.0 31.0 6.8 27.2
125 3.5 73.2 2926 36.6 -999.9 15.0
125 1.2 52.6 15.4 2964 -99909 12.5
125 1.2 48.9 14.2 28.5 -999.9 10.7
125 1.2 52.2 15.6 29.2 -999.9 11.9
0 1.3 19.0 726 27.6 9.3 34.7
tt) 1.1 16.7 668 25.9 11.9 35.0
i) 1.2 17.7 72 25.9 9.2 35.1
1 1.2 24.8 10.6 33.6 15.4 38.1
1 1.3 25.4 11.8 33.5 16.8 42.3
1 1.2 22.8 92 31.3 15.7 41.6
1 1.2 23.6 10.9 29.7 8.9 42.5
1 oo 20.3 902 24.7 8.1 37.3
1 1.1 22.4 10.3 29.5 a) 42.4
5 1.1 7202 2262 77.7 16.6 50.4
5 1.1 72.0 21.8 76.1 15.5 49.6
5 1.1 68.3 19.9 72.6 18.5 48.6
5 1.0 33.0 10.3 34.0 8.2 44.2
5 1.0 32.2 10.0 34.3 7.8 43.2
5 1.2 32.8 10.6 31.8 9.3 42.5
25 1.4 42.0 14.2 38.9 7.8 41.1
25 1.5 38.2 13.4 35.2 7.6 40.4
25 1.5 38.7 13.0 35.9 10.2 37.9
25 1.1 41.7 14.5 40.8 8.7 28.8
25 1.0 41.0 14.2 40.0 8.6 32.5
25 1.0 43.0 15.0 44.9 8.4 32.0
59 1.5 5029 18.0 33.8 17.3 49.7
50 1.2 51.1 18.0 34.5 18.3 56.2
50 1.4 52.8 17.9 33.9 18.6 51.7
50 1.2 46.4 16.4 3504 9.9 42.4
50 1.4 44.4 16.0 36.0 10.8 43.
50 1.1 43.7 15.7 34.2 12.6 38.4
85 1.7 89.8 20.6 53.0 9.9 43.1
85 1.5 90.4 18.0 50.2 9.6 41.4
85 1.7 94.9 20.3 43.5 9.4 43.1
85 1.9 73.2 19.8 54.0 6.9 32.9
85 1.8 68.0 20.7 53.3 6.5 31.4
85 1.9 20 19.8 54.0 9.9 41.4
125 1.8 78.0 18.3 36.0 3.3 33.0
125 1.8 75.8 17.8 35.5 3.3 32.9
125 1.9 80.4 18.7 37.2 2.9 32.3
125 1.6 60.6 14.1 37.2 Tel 21.3
125 1.8 68.6 15.2 40.0 7.7 21.8
125 1.6 67.0 20.7 ot) 7.8 19.2
0 2.1 7.2 667 15.6 20.2 24.3
0 202 723 6.5 16.0 17.8 27.9
0 2.2 7.6 605 16.5 17.4 29.6
1 2.0 8.9 7.8 20.1 18.2 24.1

88


£6

CLAM
DAY NUMBER
CADMIUM Tx 348
CHROMIUM T= 1.20
COPPER
NICKLE
LEAD
ZINC
CADMIUM f= BB
CHROMIUM T= 38
COPPER
NICKLE = T= 71
LEAD Ts ll
ZINC Te 88
DAY NUMBER
CADMIUM Te +04
CHROMIUM
COPPER
NICKLE = T= 203
LEAD Te ae
ZINC Te 2
DAY NUMBER
CADHIUM T= +72
CHROMIUM T= 1,00
COPPER
NICKLE T= 39
LEAD Tz +43
ZINC Ts 1.28
NUMBER
ADM 1UM +33
CHRONIUN +96
COPPER +00
NICKLE = T= 3624
LEAD Ts 97
ZINC Te 1.01
DAY NUMBER
CACKIUM T= 82
CHROMIUM Te 2.47
COPPER = Te
NICKLE = Te
LEAD Ts
ZINC Te

SHELL

0
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN=ChEAN
NOT SIGNIFICANT
NOT SIGNIFICANT
NUT SIGNIFICANT
1
NCT SIGNIFICANT
NOT SIGNIFICANT
EMEAN =CMEAN
NDT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT

5
WOT SIGNIFICANT
EMEAN=CHE AN
EMEAN =CHE AN
NOT SIGNIFICANT
NGT SIGNIFICANT
NOT SIGNIFICANT
25
NOT SIGNIFICANT
NOT SIGNIFICANT
EMEAN SCRE AN
NET SIGNIFICANT
NOT SIGNIFICANT
NGT SIGNIFICANT
50
NOT SIGNIFICANT
WOT SIGNIFICANT
NOT ’STGNIFI CANT
SIGNIFICANT
NUT SIGNIFICANT
NOT SIGNIFICANT
125
NOT SIGNIFICAKT
NOT SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICAKT
NOT SIGNIFICANT

EXPERIMENT NO=1

EMEAN-CME AN=
EMEAN-CME Al

EME AN-CME Al
EME AN-CME Ar
EME AN-CME AN =

EME AN-CME AN=
EME AN-CHE AN=

EME AW-CME AN=
EWE AN-CME Biv =
EME AN-CME AN =

EME AN~CME AN=
EME AN=CME AN=

EME An-CME AW =
EME AN-CME Al

EME AN-CMt AN=
EME AN~CME An =

EME An~CME Al
EME AW-CME Al
EME AW-CME AN =

EME AN-CME Al
EMEAN-CME AN
EME AN-CME AN =
EME AN-CME AN =
EME AN-CME AN=
EME AN-CME AN =

EME Ane CME ANS
EME AN~CME AN=
EME AN-CME Al
EME Aw~CME Al
EME An~CME AN =
EME AW=CME AN =

215
1.67

~225
-1.80
“1.27

1ell
+73

9.19
1.68
1.12

+05

OEG=
DEG=

DEG=

DEG=

DEG=
DEG=

LEGS
DEG=
DEG=

DEG=

ee coe 2 woe oo

BE ESEES COW

pppens

EMEAN=

EMEAN=

3.60
3.37
G45
16.90
30.20
2.35

2.74
5.02
4645
26.34
34.42
3.45

1.82
1.70
4.45
16.75
28.70
2.67

2.86
2.22
445
22.92
32.47
4.07

4.30
2465
4.45
32.45
26.95
14.62

4.35
2.20
4.45
33.20
47.10
4.07

EDEV=
EDEV=
EDEV®
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=

EDEV=

EDEV=

EDEV=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
eDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EOEV=
EDEV=
EDEV=

47
2.89
2.89
19.00
16.98
Ael2

1.86

2.32
2.32
17.31
24.31

1.68

1.94
1.94
1.94
18.82
15.36

1.50

1.90

1.05
1.05
19.46
19.53

2.05

81
Aeld
200
1.75
17.93
10.77

1.al
1.00
00
043
2.80
2.31

CMEAN=
CMEAN=
CHEAN=
CMEAN=

CHEANS
CHEANE
CHEAN=
CMEAI
(MEAN
CMEAN=

CMEAN=
CMEANS
CMEAN=
CHEAN=
CMEAN=
CHEANS

CMEAN=
CMEAl
CMEAN=
CMEAN=
CMEAN=
CMEAN=

CMEAN=
CME AN=
CMEAN=
CNEAN=
CMEAN=
CMEAN=

CMEAN=

CHEAN=

2

3.45 CDEV=
1.70 CDOEV=
4.45C0EV=
17.15 CDEV=
32.00 COEV=
3.62 CDEV=

1.62 CDEV=
4.30 CDEV=
4.G5CDEV=
17.15 COEV=
32.55 CDEV=
2.33 COEV=

1.77 COEV=
1.70CDE
4.45CDEV=
17.10 CDEV=
30.15 CDEV=
2.65 CDEV=

1.67 COEV=
1.70 CDEV=
4.4SCDEV=
17,15 COEV=
26.52 COEV=
2.52 CDEV=

4.50
3.45
4.45
36.70
13,85
6.45

COEV=
CDEV=
CDEVe
CDEV=
COEV=
COEV=

5.10
4.20
HGS
30.95
44.95
3.10

COEVE
CDEVe
CDEV=
CDEV=
COEV=
COEVs

41
+00
200
19.28
23.27
1.68

1.70
3.00
00
19.28
21.90
1.62

1.88
200
00

19.23
21.54
1.46

1.99

00
19.28
19.43

1.30

“14
207
+00
2G
1.63
2.05

+28
71
200
1.34

014


60T

URCHIN
DAY NUMBER
CADMIUM T= 14
CHROMIUM T= 7.54
COPPER = T= 3.56
NICKLE T= 10.58
LEAD Ts 3.06
ZINC Ts 49
DAY NUMBER
CADMIUM Ts 1.00
CHROMIUM T= +16
COPPER = T= 1.80
NICKLE 2.03
LEAD Ts
ZINC Ts 8.64
DAY NUNBER
CAOMIUK T= 2.04
CHROMIUM T=
COPPER = T=
KICKLE f=
LEAD Ts
ZINC Ts
DAY NUMBER
CADMIUK T= Bt)
CHROMIUM T= 4.78
COPPER = T= 3.39
NICKLE = T= 6.16
LEAD Ts 16.72
ZINC T= 12.75
DAY NUMBER
CADMIUK T= 3.43
CHROMIUM T= 1.82
CGPPER = 6642
NICKLE T= 2.73
LEAD Ts 10.95
ZINC T= 7.51
DAY NUMBER
CADMIUM T= 1.28
CHROMIUM T= 11.99
COPPER = T= 7.59
NICKLE 2.51
LEAD 19.08
ZINC T= 7.24

INTERNAL

0
NOT

SIGNEFICANT

SIGNIFICANT
SIGNIFICANT
SIGiWIFICAT
SIGwIF ICANT

NOT
1

WOT
NOT
not
att
NOT

SIGNIFICANT

SIGNIFICANT
2TGNIFICANT
SIGNIFICANT
STONTFICANT
SIGNIFICANT

SIGNIFICANT

5
NOT

STGNIFIL CANT

SIGHKIF ICANT

NCT

SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

25
NOT

SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

50

SIGNIFICANT

Not

SIGNIFICANT

SIGNIFICANT

woT

SIGNIFICANT

SIGNIFICANT

125
NOT

SIGNIFICANT

SIGNIFICANT

SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGWIFICANT
SIGNIFICANT

EXPERIMENT NO=1

EME AN-CME ANS
EME AN-CME ANE
EME ANCHE AN ®
EME AN-CME Al
EME AN-CME AN
EME Au-CME aN =

EME AN-CNE AN=

EME Aw-CME AN =
EME AN-CME ANS
EME AN-CME ANS
EME AN-CME AN =

EME AN=-CME AN
EAE AM-CNE AN
EME AN-CME AN =

EME Ad-CMc Ane
EME Aid-Ci4E ANE
EME Aw-CME AW =
EME AW-CMc AN =
EWE AN-CME AN®
EME Aiv-CME AN=

EME AN-CME AN =
EME Aw-CME AN=
EME AN-CME Ai
EME ANW-CME AN =
EME AN-CHE N©
EME AN=CME AN =

EMEAN-CME AN®
EME AN=CNE AN =
EME AN=CHE AN=
EME AN-CME AN =
EME AN-CME AN =
EME AN-CME AN =

29.92
69.26

vDEG=
bEve
DEG=
vEGS
DEu=
DEG

LEGS
DEG=
DEGs
btGe
ves
DEG

bEus
DEG
bEG=
DEGs
DEG=
DEGs

DEGs
DEG:
LEG?
DEG=
DEGe
Deus

DEG
DEG=
DEG
DEG:

G DEG=

vEG=

DEG:
DEG=
vEG=
DEG=
DEG:
veGs

ANNAN RET EES

eppeses

SHUEOE

C@awwe NREEES

EMEAN=
EMEAW®
EMEANS
EMEANS
ENEANS
cMEANS

EMEbAE
EMEAN=
EMEAN=
EMELNS
cM AW=
EMEAN®

EMEA
EMEANE
EMEAN=
EmMEANS
EMEANS
EMEANS

EMEAN=
EMEGN=
EMEANS
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEANS
EMEAN=
EMEAN=
EMEANS
EMEAN=
EMEANS

2426
3.20
5.56
16.42
13.62
46240

«1S
9.85
11.65
21.46
17.75
61.20

1.07
12.90
13.90
39.75
28.20
72.06

4.37
24.97
22.00
39.55
38.66
90.90

2.30
36.50
25.95
55.85
31.10
94.715

3.82
20.32
21.20
22.83
50.27

160.73

EOEV=
eDEV=
EDEV=
EOEV=
tDEV=
EDEV=

tOEV=
EDtV=
Eveve
eDEVe
EDEV=
EVEV=

EDEV=
cDEV=
bocve
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDtV=
EDEV=
EDEV=

EDEV=
EvEV=
eDEV=
EDEV=
EOEV=
EDEV=

EDEV=
EUEV=
EDEV=
EDEV=
EDEV=
EDEV=

1298
1.76
2.22
2.48
1.00
481

00
064
207
00
2.33
2.69

1.31
1.41
2.83

207
1.13
3.39

1.50
5.02
2.76
459
1.31
3.51

14
214
1.46
2.90
2.26
2.90

1.07
1.92
3.63
3.86
3.04
14.83

CHEAN=
CHEAN=
C4EANS
CMEAN=
CAEANS
CHEAN=

CHEANS
CREAN=
CMEANS
CHEAN=
CMEAN=
CMEAN=

CMEANS
CMEAN=
CMEAN=
CMEAN=
CMEAN=
CMEAN=

CMEAN=
CMEAN=
CMHEANS
CMEANS
CHEAN=
CMEAN=

CN EAN=
CMEAN=
CMEAN=
CMEAN=
ChEANE
ChEAN=

CHEAN®
CMEAN=
CHEAN=
CHEANS
CMEAN=
CMEAN=

gy

2.05
15.60
11.60
36.40
17.95
48.15

1.92
9.75
7.02
25.15
19.55
44.710

3.45
11.07
9.16
26.65
17.60
35.72

4.57
11.72
13.72
22.70
16.70
47.40

4.02
31.70
18.92
47.75
20.10
77.35

4.85
7.75
5.59
17.67
20.35
411.467

CDEV=

e2l
2.26
71
7h
2.76
07

2.51
264
3.64
2.62
1.20
+28

1.35

31
3.50
3.68
2.21
3.28

1.42
2.35
4.02

+70
1.94
5.85

267
3.53
1.31
3.56

229
0.071

1.51
295
2.28
1648
57
14.61


ZN MANGROVE ROOT— E&xP 2

-2

MEAN <E-C> /PPM

—4

4

a
@h
28
az! 7

DAYS

ZN THALASS |R ROOTS— ©&XP. 2

=4
=
i if 4
S
a
Ye
w
y
z
ced
i
=
“= r T T T T T T 1
s & & z
pRAYS

Figure 36. Mean net uptake of Zn between experimental and control,
Thalassia roots and mangrove roots - Experiment II.

56


PB HOLOTHURIAN GUT- EXP. J

so J
=
Q 4
a
~
up J
“a
ng
li
~ 221
z
a 4
=o J
— T T , T —
ss = o -
5 5 u
a
CRYS
ZN HOLOTHURIAN GUT- EXKP. t
so J
= 4
£ up 4
nA se]
y
Wo ze
= 4
i id
lJ
= 4
a
T T T T t 7 ——
= F 8 5
CAYS

Pigure 42. Mean net uptake of Pb and Zn between experimental and
control, sea cucumber guts - Experiment I.

62


All other equipment was washed with detergent, rinsed with
R.0O./D.I. water, followed with 6N HC& and rinsed with R.0O./D.I.
water. Only non-metallic instruments and materials were used
and all equipment , after cleaning, was stored covered with
Parafilm or plastic bags.

Oxygen, temperature, and salinity

A Yellow Springs International YSI - 57 oxygen/temperature
meter was used. The oxygen output was both salinity and temper-
ature compensated. The oxygen calibration was checked using the
Winkler titration technique (Strickland and Parsons, 1965).

Dissolved reactive phosphate

One hundred m& samples were collected and immediately
filtered through a Millipore Swin-Loc adaptor containing an
acid washed (6N HCL) 27 mm Millipore, 0.45 m pore size filter
using an acid washed 100 m& polypropylene syringe. The sample
was frozen and later DRP was determined using the single
solution technique (Strickland and Parsons, 1965) with a 10 cm
cell in a Beckman DU spectrophotometer. A small number of samples
were run on a Technicon autoanalyzer using standard Technicon
procedures (Zimmermann, C. et al., 1977).

Particulate trace metals

The 142 mm diameter filters were placed in Pyrex R beakers,
6 m& of concentrated HNO,: HC& (3:1) were added and the beaker
covered with a ribbed watch glass. The solution was heated to
a slow boil (less than 95°C), when the filters dissolved, the
liquid was brought to a rapid boil and taken to near dryness.
Then, 4 m& concentrated HNO, were added, followed by dropwise
addition of 1 m& of H,0, (30% by volume). The sample was then
reboiled to near drynéss and 3 drops of concentrated H $0, were
added. If the sample turned black, more H,0, was added. The
samples were dried until SO, fumes disappearéd. The sample was
then brought to 10 m& volumé with R.O./D.I. water.

Soluble trace metals

After 2.52 of sample was filtered for determination of
"particulate" trace metals (41 in Experiment II), the effluent
was run through two 1.2 cm diameter x, 10 cp glass columns. The
flow rate was maintained at 1 mg min em“. The columns were
in series and the first column contained 10 m& of Chelex-100
in the ammonium_form (20 m& in Experiment II) with 20 m& of
Amberlite XAD-2® resin (Rohm and Haas) in the second column.
The Chelex resin was purified by 2 bed volumes (BV) of 1NHC2
(isothermally distilled) followed by 5 BV of D.I./R.O. water
and then 2 BV 1N NH,OH (isothermally distilled) with a final
wash of D.I./R.O. water. The Amberlite XAD-2 resin was cleaned

8


ATAM OH ORBNHOMNnH GT COOO1HO

Cs eee er ee oe eee eee

MUTNDOR OER HAtTTAMH Orr OOO

OEE NTNHOTDOMPADDOMOWOS

AAAIBNOM OBE EHAOOTMNNE®

Rr eee ere ar ee ee re
ANNAN ANNANN ANNAN NN

SORTED DATA
1.9

25

WWINOODWASO CAN inn in
NANNKNNNNNODDWODMAN

125
125
125
125
125
125

OF AH DOOMHAHOOO HAH ODO MM
NNN NNNN NANA ANNAN NNN
ANMNNNMMNNMNANMNMNMNNMNMNNNNM
P9273 PIV A273 3T9I972D99N9N 79399092

90


of the sampling period (85 days), For Experiment II, Cd and Ni
demonstrated a steady uptake until day 50 (Figs. 7 and 13) then
a decline in the net uptake. Zinc and chromium indicated a net
uptake after day 50 (Figs. 12 and.15) and Cu showed little sig-
nificant uptake for the sampling period (Appendix B).

Sewage sludge

The initial values for trace metals in sewage sludge are
shown in Table 4.

A rapid decline was noted in the concentration of Cd, Cr,
Cu, Ni, and Zn in the sewage sludge for Experiment I (Figs. 6, 15,
8, 14, 11). The Pb results are inconclusive for both experiments
(Fig. 9 and Appendix B) due to the loss of lead as the easily
volatilized PbSO, during the digestion of the sludge. Because
of the shorter s4mpling interval for sludge in Experiment II, a
two stage decline for Cd, Ni, and Zn was shown (Figs. 7, 13, 12).
The first decrease in metal concentration was nearly instantaneous
with a further, smaller decrease after day 50.

Thalassia
Leaves

The results of EXP I were biased due to the "die off" of the
plants by day 50. This was probably due to insufficient light.
The plants survived for the full 125 dav period in Experiment II
after the light level was increased.

Copper, Cr, Pb, Ni and Zn showed rapid increases in net
uptake in Experiment I (Figs. 16, 18, 20, 22, 14). In Experiment
II the same overall trend of increasing net uptake was shown for
Cu, Cr, Pb, and Cd for Experiment II (Figs. 17, 19, 21, 26).
Nickel and zinc, in Experiment II did not show a significant
uptake up to day 50 but the net uptake was significant at day 85
and 125 (Figs. 23, 25).

Root

The only significant net uptake of trace metals for Thalassia
roots and rhizomes in Experiment I was for nickel, Cr, Cu and Pb
(Figs. 27, 29, 31, 33).

The repeat experiment also showed a significant net uptake
for Cr and Zn (Figs. 30, 36) with Pb, Ni, and Cu showing peaks
of significant net uptake values at day 5 and/or 25 days (Figs. 34,
28, 32). Cadmium uptake was significant only at day 25 (Fig. 37).

Urchin, internal organs

To facilitate later discussion, the results of the net uptake
for urchins are shown with the results for Thalassia leaves.

13


PB THALASS (IA LERVES—- ©&xXP. 2

=e
Qa
a
ro
~ 5s 4
Vv
w
wi |
z
fia
Ww 2 4
=
a T TT T T T Te a
s & & uw
8
caAYS
PBS URCHIN GUT- EXP. 2
ue |
= 4
+ ae 4
a 4
Yo ae 4
ir)
Ww 4
r4 im
a
= 4
eo 4
SELES ee
= t T
a Fy
DAYS

Figure 21. Mean net uptake of Pb between experimental and control,
Thalassia leaves and urchin guts ~ Experiment II.

41


vov00ev00nD00N000

ANUNUHUUNUUNUUWUNUNUHNUDDRDDDOHDDDDDODDRDDHDDHMHVVUVVTVVVVVUVUVUUU
NNN NNNRN NNN NNR NNN NNNNNNNNNNNNNYNNNNNNNNN NN NNNNN NYDN
HKPOOOM MB ODOKP HE HM ODOODDDOOODODDDDDOQCDDO OOOO OPH Hr ERB OOOrHHOOO

SORTED DATA 17

50 2.0 498.0 55.9 327.7 44.0 88.9
50 2.0 532.3 55.4 357.1 45.6 86.1
50 2.3 539.0 58.8 361.6 20 75.0
50 . 164 429.6 50.8 284.0 10.6 75.3
50 1.6 437.0 64.1 293.1 10.7 90.1
50 1.8 425.1 60.2 300.0 13.8 73.8
8s 1.9 375.9 86.5 270.5 83.5 83.1
85 1.9 395.3 86.1 267.0 81.2 82.8
85 1.9 390.1 85.6 266.7 82.1 85.0
85 2.3 270.4 76.2 235.64 29.7 47.9
85 1.7 188.5 69.3 214.0 29.9 43.9
85 1.8 232.5 79.3 242.2 30.8 51.4
125 1.9 218.2 46.3 226.5 20.8 62.3
125 2.0 204.1 44.1 225.5 19.4 57.7
125 1.9 229.9 47.8 231.0 19.9 62.4

0 264.6 3267.6 2249.8 364.3 19.6 2446.7
te) 118.9 3774.8 2748.8 373.2 25.2 2648.2
0 114.4 3938.2 2337.4 389.3 19.4 2427.9
1 57-7 1449.5 1310.5 276.6 19.8 1151.2
1 50.6 1374.4 1230.9 244.6 26.4 1065.0
1 66.7 1878.8 1581.8 317.0 16.9 1365.2
5 63.3 2022.2 994.6 328.7 22.9 1463.1
5 53.3 1763.2 1058.4 307.0 16.9 944.0
5 60.3 2152.9 995.5 337.5 14.7 1377.6
25 56.4 1993.1 1185.6 254.0 49.3 1378.7
25 45.5 1492.6 1652.7 224.3 32.5 933.1
25 52.8 2052.2 1082.1 258.0 20.3 1305.1
50 62.0 2057.0 1445.3 233.7 61.7 1474.0
50 61.4 1815.3 1414.3 21564 58.7 1360.0
50 76.1 2064.9 1489.3 239.7 51.1 1499.7
85 40.2 1290.6 888.5 254.4 66.5 914.0
85 39.0 1239.7 873.9 239.6 70.0 879.4
85 23.2 1230.1 891.2 219.4 96.2 778.1

125 16.8 668.1 522.3 139.1 19.9 436.0
125 18.8 653.9 547.2 149.1 19.9 451.6
125 18.3 654.2 438.0 130.5 19.9 406.6
10) 6 6.6 6.7 7.2 6.6 16.2
te) 6 6.8 6.6 7.2 6.6 16.0
0 36 Tel Tel 7.5 6.7 16.2
0 6 7.5 5.7 906 7.6 15.9
0 6 7.1 6.0 9.5 7.6 16.3
0 26 6.9 5.8 9.0 8.0 16.4
1 6 6.0 604 bee Te2 15.0
1 6 5.5 6.3 6.1 6.7 15.0
1 5 5-6 : 664 6.1 6.9 15.6
1 06 7.0 6.7 Be.2 6.7 16.9
1 5 6.8 7.4 564 6.8 15.6
1 6 Tel 6.5 5.3 6.5 15.3
5 3 5.2 6.3 4.3 5.0 12.0
5 3 5.9 Tel 4.0 5.5 13.6
5 22 5.9 6.2 4.6 5.2 16.4
5 3 6.8 6.4 6.6 6.9 12.7

86


DR, CONTROL EXP. '

.S.5
44
ys
~ |
c
th 124
id
bs
5 & g Fi
CAYS
CORP, CONTROW E&XF. 2
.2 4
7 4
ae 4
E = 4
n
fia} 4
= .3 4
: 4
re
= o -
Ss i) y
DAYS

Figure 4. Dissolved reactive phosphate concentration of
Experiment I and II, control. Data points are
Mean values for 5 day period, excluding day 1.

24


vet

FOULING ORGANISMS

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
rai la

CADMIUK
CHROMIUM
COPPER

CADM 1UH
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM LUM
CHROMIUM
COPPER
WICKLE

DAY NUMBER
a 6.36
Ts 2019
tT 1.71
Ts 3.74
Ts 18.32
Ts 63

DAY NUMBER
Ts
Te
Ts
tT
1s
Ts 54

DAY NUMBER
Te 18
Ts 6.43
Te 3.75
Ts 4.37
Ts 69.53
Te 15.79

DAY NUMBER

NO EXPERMENT HEAN®

25
SIGHIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT

50
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
NOT SIGNIFICANT
65

NOT SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

125

EXPERIMENT NO=2

EME AN-CME AN
EME AN-CHE AN =
MEAN-CHEAN ©
EME AN-CHE AN®
EME AN-CHE AN =
EME AN-CME AN

EME AN-CME AN =
EME AN-CHE AN
EME AN-CME AN=
EMEAN-CME AN=
EMEAN-CHE AN®=
EME AN-CHE AN

EME AN-CME AN=
EME AN-CME AN®
EME AN-CMEAN®
EME AN~CHE AN=
EMEAN-CMEAN=
EMEAN-CHE AN®

1.93 DEVIATION=

NO EXPERMENT MEAN®= 217.40 DEVIATION=

WO EXPERMENT HEAN®

46027 DEVIATIONS

NO EXPERMENT MEAN= 227.67 DEVIATION=

NO EXPERMENT MEAN®
NO EXPERMENT HEANS

20.03 DEVIATION=
60.80 DEVIATION=

DEG=
DEG=
DEG=
DEG=

23.73 DEG=

DEG=
DEG=

92.53 DEG=

DEG

56.43 DEG=
33.10 DEG=
3.60 DEG=

DEG=
DEG:
OEG=
DEG=
DEG=
DEG=

NUM®
NUM®
NUKS
NUM=
NUM
NUN®

VELLOw

PERS ES

PoRSetr

REEDED

EMEANS
EMEAN=
EMEAN®
EMEAN=
EMEAN=
EMEANS

EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN=
EMEAN®

EMEAN=
EMEAN®
EMEAN=
EMEAN®
EMEAN=
EMEAN=

1.83 EDEV=
445.23 EDEV=
49.13 EDEV]
337.10 EDEV=
37.07 EDEV=
75.50 EDEV=

EDEV=
0 EDEVs
EDEV=
EDEV=
44.80 EDEV=
83.33 EDEV=

1.90 EDEV=
367.10 EDEV=
@6.07 EDEV=
268.07 €DEV=

206 CHEAN®
51.53

17.87
3.36

4
18.41
1el3
7235 CHEAN=

«00 CHEAN=
10.04 CHEANS
45 CHEAN=
old CHEAN®
1.16 CHEAN®
1.19 CHEAN=

206


by washing for 10 minutes with D.I./R.O. water. The water was
decanted and the procedure repeated 5 times. The resin was
washed four times with anhydrous methanol followed by four

washes with D.I./R.O. water. The Chelex resin was left covered
with water prior to use. The Amberlite was stored under methanol
until just prior to use.

In Experiment I and II, the metals were extracted from the
Chelex-100 ® resin after the resin column was first washed with
100 m& of water. The trace metals, in EXP 1, were eluted with
40 m& of boiling concentrated HC2£. The resin was rinsed with
20 mg of water and this water was combined with the acid eluate.
The eluate was boiled down (95°C) to 1 m& and then brought to
10 mg volume with water. This was referred to as the soluble
inorganically complexed fraction. In Experiment II, the Chelex
100 was eluted batchwise using 6 m& concentrated HNO, per 10 mg
resin (Davey, E.W. and A.E. Soper, 1975) rather than~eluting
with concentrated HC&. The Chelex resin was filtered and the
acid eluate dried to 1 m& followed by the addition of 6 mg
2NHC2 (isothermally distilled) and dried down to 1 m&. This was
repeated and the final product was diluted to 10 mg with distilled
water. The Amberlite vesin was first washed with 100 m& water
then the resin was placed in an open glass Petri dish, rinsed
with methanol, and the resin ignited. The ignited resin was
placed in a muffle oven and the temperature raised in 100°C
increments until 400°C. The resin was maintained at 400°C for
1 additional hour then the temperature was raised to 450°C for
24 hours. The cooled resin was dissolved in 5 m& iINHC&, then
boiled down to 1 m% and brought to the desired final volume.
This fraction was referred to as the soluble, organically complexed
trace metal (Montgomery, J.R. and J.E. Echevarria, 1975). The
trace metals were determined by atomic absorption spectrophotometry
(A.A.S.) on a Perkin-Elmer Model 303 A.A.S with a deuterium back-
ground corrector. ‘The output was recorded using a Model 56 Perkin-
Elmer recorder and DCR-1 digital readout. The flow chart for
sample collection and processing is shown in Fig. 3.

Biological and sediment samples

Analyses of National Bureau of Standards (NBS) bovine liver
and orchard leaves were performed along with the regular samples.
One gram samples were used, when available, for all standards
and samples (see Table 3). The flow chart for sample collection
is shown in Figure 3.

The number of organisms sampled per day per experiment is
shown in Table 1. The samples were defrosted after collection
and dissected into the following tissues using Pyrex glass shards
and plastic gloves.

Urchins - shell and internal tissues.

Sea cucumbers - body, muscle bands and internal tissues. In
Experiment II, muscle bands were combined with internal tissues.

9


cu MANGROVE ROOT—- ©&xP. 2

is.
=.
ft tad
S
A
oe |
ug
Vv
Zz
ce J
WwW
=

-s

~ " T z T 7
Ps
DAYS

cu THALASS!|A ROOTS—- ©&xXxP. 2

is,
=
a te
S
A
Gel
Ww
v
Zz
cme 4
ld
=
-= 71

Figure 32. Mean net uptake of Cu between experimental and control,
Thalassia roots and mangrove roots - Experiment II,

52


Tet

NERITA SHELL EXPERIMENT NOs2
DAY NUMBER 0
CADMIUM NO EXPERMENT MEAN= 3.77 DEVIATIUN=
CHROMIUM NO EXPERMENT MEAN® 7.67 DEVIATION=
COPPER NO EXPERMENT MEAN= 4.45 DEVIATIONS
NICKLE NO EXPERMENT MEAN= 12.10 DEVIATION®
LEAD NU EXPERMENT MEAN= 36.00 DEVIATIGN®
AINC NG. EXPERMENT MEAN= 4.97 DEVIATI UNS
DAY NUMBER 1
CADMIUM T= 1.79 NOT SIGNIFICANT EMEAN-CMEAN®
CHRDMIUM T= +71 NOT SIGNIFICANT EMEAN-CMEAN®
COPPER EMEAN =CME AN
NICKLE Ts 1.75 NOT SIGNIFICANT EMEAN-CMEAN=
LEAD Ts +51 NOT SIGNIFICANT EME AN-CME AK =
ZINC T= +23 NOT SIGNIFICANT EME AN-CHE AN®
DAY NUMBER 5
CADMIUM T= +36 NOT SIGNIFICANT EMEAN-CME AN?
CHROMIUM T= 1.21 NOT SIGNIFICANT EMEAN-CMEAN®
COPPER EME AN =CME AN
NICKLE Te +53 NOT SIGNIFICANT EMEAN-CME AN®
LEAD Ts 265 NOT SIGNIFICANT EMEAN-CME AN
ZINC Te 06 NOT SIGNIFICANT EME AN-CMEBN®
DAY NUMBER 25
ADM 1UM EMEAN =CME AN
CHROMIUM T= 2-04 NCT SIGNIFICANT EMEAN-CMEAN=
COPPER EMEAN=CKHEAN
NICKLE = T= +21 NOT SIGNIFICANT EMEAN-CME AN =
LEAD Te +63 NOT SIGNIFICANT EMEAN-CMEAN®
ZINC Te =26 NOT SIGNIFICANT EMEAN-CME AN®
DAY NUMBER 50
CADMIUM T= 246 NOT SIGNIFICANT EME AN-CHE AN=
CHROMIUM T= 262 NOT SIGNIFICANT EME AN-CME AN]
COPPER Te 280 NCT SIGNIFICANT EMEAN-CMEAN=
NICKLE Te 1.76 NOT SIGNIFICANT EMEAN-CMEAN@
LEAD Ts 5.23 SIGNIFICANT EME AN-CME AN =
ZINC Ts +38 NOT SIGNIFICANT EME AN-CMEAN=
DAY NUMBER 65
CADMIUM T= 2.12 NOT SIGNIFICANT EME AW-CNE AN®
CHROMIUM T= +76 WGT SIGNIFICANT EME AN-CMEAN®
COPPER T= +44 NOT SIGNIFICANT EMEAN-CMEAN®
NICKLE Te °81 NOT SIGNIFICANT EMEAN~CMEAN=
LEAD Te +80 NOT SIGNIFICANT EME An
ZINC Te 1.87 NOT SIGNIFICANT EME An
DAY NUMEER 125
CADMIUM NO CONTROL MEAN= 2645 DEVIATION=
CHROMIUM NG CONTROL MEANS 9.63 DEVIATIUN=
COPPER wO CONTROL MEANS 4.45 DEVIATIUN
NICKLE NG CONTROL MEAN= 12.67 DEVIATIU
LEAD NO CONTROL MEAN® 42.83 DEVIATIUN®
1INc NG CONTROL MEAN= 4.80 DEVIATION=

045

290 DEGe

7213 DEG=
-260 DEG=
210 DEG=

7.07 DEG
+33 DEG=
+33 DEG®

Deus

2.20 DEGs

OEGs

DEG
DEL
DEG
DEG=
DEG
DEvs

z
€
z
.
euvwow

NUh= 3

NUM=
NUMS
NUM
NUKa
NUM®

vueww

+ PDR PE SEE FE

ES

Prepare peapes

EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEaN®
EMEAN®

EMEAN®
EMEAN=
EMEAN=
EMEAN®
EMEAN®
EMEANS

EMEAN®
EMEAN®
EMEAN®
EMEAN=
EMEAN*
EMEAN®

EMEAN®
EMEAN=
EMEANS
EMEAN®
EMEAN=
EMEAN®

EMEAN=
EMEANE
EMEAN®
EMEANS
EMEANS
EMEAN=

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEVs

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=

EDEV=
EDEV=
EDEV=
EDEVe

EDEV=
EDEV=
EDEV=
EDEV=
EDEV>
EDEV=

EOEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

232

+06
+36
215
15
46
So

CHEANS
CMEAN=
CMEANS
CHEANE
CMEAN=
CHEAN=

CHEANS
CHEAN=
CMEAN=
CHEAN®
CMEAN=
CMEAN=

CHEAN=
CMEAN=
CHEAN
CMEAN=
CHE ANS
CHEANS

CHEAN=
CMEANS
CHEAN=
CHEAN=
CHEANS
CHEAN®

CHEAN=
CMEAN=
CHEANS
CHEANE
(MEANS
CHEAN®

2

3-47 CDEVe
6.50 CDEV=
4-45CDEVE
13.00 COEV=
36.50 CDEV=
5.47 COEVE

3-97 CDOEV=
7.07 COEVe
44S5CDEV=
12.20 COEV=
33.60 CDEV=
4.47 CDEV=

3.1 7CDEV=
8.53 COEV=
4.45CDEV=
11.67
33.23
4.43 COEV=

3.20 CDEV=
7.87 COEV=
9.90 CDEV=
15.37 CDEV=
46.50 CDEV=
4.57 CDOEV=

2.33 COEV=
8.93 CDEV=
9.40 CDEVe
13.73 CDEV=
48.93 CDEV=
4.40 CDEV=



SOT

FOULING ORGANISMS

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

DAY NUMBER
T= 00
Ts +00
Ts +00

. 200

= 209
Ts +00

DAY NUMBER
Ts 15.01
tT 82
Te 19.23
T= 1182.41
Ts 29.73
T= 22.96

50

NOT SIGNIFICANT
NGT SIGNIFICANT
NOT SIGNIFICANT
NOT SIGNIFICANT
NGT SIGNIFICANT
NOT SIGNIFICANT
125

SIGNIFICANT
NOT SIGNIFICANT
SIGHIF ICANT
SIGNIFICANT
SIGNIFICANT
SIGNIFICANT

EXPERIMENT NO=1

EME AN-CHE AN
EMEAN-CME AN =
EHE AN-CHEAN =
EME AN-CME AN =
EME AW-CME AW =
EME AN-CME AN =

EMEAN-CME AN =
EME AN-CHE AN =
EHEAN-CNE AN=
EME AN-CME EN=
EME AN-CHE AW =
EME Ay-CME AN=

71.90
129.90
27.30

7.70
27.10

2.60
+10
+30
-2.00
10630
161.05

DEu=
Devs
DEG=
vEu=
DEG=
dEG=

DEG=
DEus
DEG=
VEG
DEG=
DEG=

eooo°°

forererenene)

EMEAN=
EMEAN=
EMEANS
ENEANS
EMEA
EMEANS

EMEANS

1.70
209.10
82.90
306.70
13.60
109.60

4-30
42.70
O
0

27.60
224.70

EDEV=
EDEV=
EDev=
eDEV=
EDEV=
EDEVS

EDEV=
EDEV=
EOEV=
EDEV=
EOEV

EDEV=

CHEANE
CHEANS
CHEANS
CMEANS

CMEAN=
CMEAN=
CMEANS
ChEAN=
CMEANS
CHEAN

COEV=
COEV=
COEVe
CDEVE
CDEVe
COEVE

COEVe
CDEV=
COEV=
CDEV=
CDEV=
COEV=

+00
+00
+00
+00
+00
200

214
4.10
1.41

+00

28
5.73


LOT

THALASSL

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADMIUM
CHROMIUM
COPPER
NICKLE
LEAD
ZINC

CADM TUN
CHROMIUM
COPPER
KICKLE
LEAD
ZINC

ADM TUM
CHROMIUM
CLPPER
AICKLE
LEAD
ZINC

CaAbM TUM
CHRUMIUM
CUPPER
NICKLE
LEAD
ZINC

° EXPERIMENT NO=1

MEANS 1.65 DEVIATION
MEANS 1.70 DEVIAT Ons
MEAN® 14.03 OEVIAT IbWe
MEAN? 9.05 DEVIATIUN=
MEANS 11.50 DEVIATIONS

MEANE 36.63 DEVIATION=

1

233 NOT SIGNIFICANT EMEAN-CME AN =

LEAF
OAY NUMBER
NO EXPERMENT
NO EXPERMENT
NG EXPERMENT
ND EXPERMENT
NO EXPERMENT
NU EXPERMENT
DAY NUMDER
Ts
T 3.11
Te 10.76
Te 2.99
Ts 12.16
Te 4.65

DAY NUMBER

2619 NOT SIGNIFICANT

Ts +50 N
ie 2.51 N
Te G95
Ts 1.862 N
Te 3.86
Ts 7.78
DAY NUMBER
Te «BS Ni
Ts 4.55
Ts 3.2

Ts

Te 6.87
Ts 3.31
DAY NUMBER
Te 481
Te 15.48
Ts 7.01
Ts 4.29
Ts 22.24
Ts 9424

SIGNIFICANT
SEGNIFEChnT
SIGNIFICANT
SIGHWIF ICANT

SIGWIF ICANT
5

GT SIGNIFICANT

GT SIGNIFICANT

SIGHIFICANT

OT SIGNIFICANT

SIONTFICANT

SIGNIFICANT

25

GT SIGHIFI CANT

SIGNIFICANT
SIGNIFICANT

SIGNIFICANT
SIGWIFICANT
50

SIGNIFICANT
SIGWIF ICANT
SIGNIFICANT
SIGHIFICANT
SIGNIFICANT
SIGNIFICANT

EME AN-CKE Aa =
EME AN-CME AN =
EAE AN-CME AN =
EME AN-CMt AN =
EAE W-CAE BN =

ENE AN-CME AN =
EME AN-CME AN =
EME AN-CME AN=

EME AN-CME AN =
EME AN-CME AN =

EME Aw-CME AN=
CAE AN-CME aw =
EME AN-CME wid =
EME AN-CME BN =
ENE Aw-CME AN =
EME AN-CME AN =

EME AN-CNE GW =
EWE AV-CME AN
EME An-CME AN =
EME AN-CME AN =
EAE AN-CME AN =
EME ACME AN =

1.30 NUM= 3
+00 NUM= 3
1.53 NUM= 3
7-46 woM= 3
2.04 wUMs 3
9.32 NUM= 3

~+22 DEG=
8.18 LEGS
6.47 DELS
11.67 LEGS
17.15 DEG=
10.32 DEv=

+42 OE
7.48 VE
7-62 DEG
72.35 DEG=
10.61 DE
10.27 GE

=2.24 VEG=
33-15 vEb=
12.85 Deus
15.60 DEG?
18.80 DEG:
~4.45 DEG

5.43 DELS

27.90 VE
77.08 veGs
25.90 DEGs

WNL UNS BUFUUN KOOeee

RUbous

EMEANS
EMEANS
EMEANS
EMEAN®
EMEAN=
EMEAN=

ENEAN=
EMEANE
EMEAN=
EMEAN=
EMEAN=
EMEAN=

EMEAN=
EMran=
EMEAN=
ENEANS
EMEAN=
EMEANS

EMEANS
EMEGN=
EMEAN=
EMEANS
EMEAW=
EMEAN=

3.00
27.65
21.27
50.15
32.62
50.30

2462
25672
22.82
44625
31.07
45.97

4.04
05.60
27.05
47.50
35,60
28.70

4670
94.60
59.73
70.70
95.33
54.70

1.05
3.81

7.28
2.30
3.74

1.6
2.57
2.05
1.72

98
1.20

442
3.11
4.71
9.50
3.92
147

1.14
5466
8.73
£OEV= 10.15
EDEV= 438
EDEV= 2.26

CHEAN=
CMEANS
COEANS
CHEANS
MEANS
CKEANS

CMEANS
CMEAN=
CMEAN=
CMEANS
CHEAN=
CMEAN®

CMEANS
CHEANS
CHEANS
CMEAN=
CHEANS
CMEANS

CMEAN®
CHEANS
CHEAN®
CMEAN®
CHE AN=
CHEAN=

17

3.22
19.47
14.80
38.47
14.87
39.97

2.20
$3.20
15.20
46.60
20.47
35.70

6.27
32.45
14.2
31.90
16.80
33.15

10.13
35.87
22.50
42.80
16.25

CDEV=
CDEV=
CDEVE
CDEV=
COEV=
CDEV=

3.00
2.05

1.59
3.15
2.91
4.90

3.25


FIGURES

Number Page
Study site location....... se eeee eee er ones see e eee eee eee 2Ta
Diagram of experimental design, tank configuration,

and sea water system.........eeeeeee deem eee eee eens 22
3 Flow chart of trace metal and hydrocarbon sampling

from collection processing into final sample

form.......... beeen eee teen e eee eee eee eee eee 23
4 Dissolved reactive phosphate concentration

Experiment I and II, control. ...... ccc eee eee eee eee 24
5 Dissolved reactive phosphate concentration

Experiment I and II, experimental........ wee ee see eee 25
6 Mean net uptake of Cd between experimental and

control, fouling organisms and sewage-

Experiment I........ eee eee eee ete eee eee ee eens eee 26
7 Mean net uptake of Cd between experimental and

control, fouling organisms and sewage-

Experiment II........ eee ee bene rece e ee eee see eeeee 27
8 Mean net uptake of Cu between experimental and

control, fouling organisms and sewage-

Experiment LT... eee cece cece eee eee eee e eee eee eee eee 28
9 Mean net uptake of Pb between experimental and

control, fouling organisms and sewage-

Experiment I.......... eee ee eee bee e eee enone re.)

10 Mean net uptake of Pb between experimental and
control, fouling organisms - Experiment II......... ++ 30
14 Mean net uptake of Zn between experimental and

control, fouling organisms and sewage-
Experiment Te... e cece cece cece eee cece cee eect cece eee 31

12 Mean net uptake of Zn between experimental and

control, fouling organisms and sewage-

Experiment II....... Se
13 Mean net uptake of Ni between experimental and

control, fouling organisms and sewage-
Experiment ID... .. cece ee cece eee cece cece eee eee eee s 33

vi


eeoeceereoec eee eo oer eee eee eee ee eer eee eee enesreserioeereoe

EOMANEONNOAST ST OBATNOTAN MME ER TAMARTOBDNANN ODEON NAAR EP aANnrovoN
oeerteroee eee eeeereoen eer esrereoeeeseeer ee eee eee eee een eee eee
DODDD ODOM DVDDDDDDDODORHOKRAHADRTOK DDN AHNNMNN OEE AOLTE DONE ST
aad a DODKHONOAHONMAKAAMFN MS

aaa ae NUN AMA

wn
AD ODRODANNKRDDDOF ROR AME DODOOMNONE NER DDOMNAMNNDOMAMNAINE ON @
cwesceeeteeeece ee ere eeseeerereereceeterereoere ere se eee tee
POR ARR ONE DORR RAR DOR OF DODAHRDROBRAHBOHROOONHHOM AST HAADANMAED
coal - aan at AN tN

ear eet ie Sr See OT
MMMM OMTMMAMMTMOMMMMAMNNNNNKDNNNANNAN NAN AMADA TAUNMTTAMMNHOM TOM

SORTED DATA

ARAMA NNN DN NNN DNODDODOCONN NH NNN NNN OOO AA AAR ANNAN NANAK
NANNANNKHANANKHNDRAODMOOVAT NNN NNN

aeaet
COMA MADDOHHADOOHAMODO RA AOOO HAH OOOH AHOOO RAB OORHOOON AAO
NUNNNNNNAN NNN NNN NNN NNN NNN NN NNN NNN NNN
DAMNMNNYDHNNYNYAYNYNDYNNHNDNYDYDYNYDNYNDYNNDNYNYD YD YY Bt ee ee et ee ee et

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84


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Figure 9. Mean net uptake of Pb between experimental and control,
fouling organisms and sewage - Experiment I.

29


OTT

ND EXPERMCNT MEAN=
NU EXPERMENT KEAN=
NU EXPEKMENT MEAN=
NO EXPERMENT MEANS
NU EXPERKENT MEANS
NG EXPERMENT MEaN=

MANGROVE
DAY NUMBER
CADM TUM
CHROMIUM
COPPER
KICKLE
LEAD
LINC
DAY NUMBER
CADMIUM T= 2.54
CHROMIUM Te +53
COPPER
NICKLE Ts 3.67
LEAD Ts 1.14
TINC Ts 1.82
DAY NUMBEK
ADK 1UK
CHROMIUM T= 3.78
COPPER
WICKLE Ts 1.12
LEAD
TING Ta 224
DAY NUMBER
CACMIUP Ts 1.39
ChROFIUN Ts 2.98
COPPER
NICKLE Te 3.06
LEAD
TIN Ts 2.39

Body EXPERIMENT NO=2

0
225 DEVIAT uke
6.87 DEVIATIONS
4.45 DEVIATIUNG
9.70 DEVIATIG
287 DEVIAT Lun
2.37 DEVIATIU

5
WOT SIGNIFICANT EMG AN-CME AN =
NOT SIGNIFICANT EME AN-CMEAN =
EME Aix =CME AN,
STGWIFICANT HE AN-CME BN
NOT STGNIFICANT ERE AN-CME An =
NCT SIGNIFICANT EMEAN-CME An =
50
EAE AN eCME RN
SIGNIFICANT
EMEA ECE aw
NOT SIUNIFICANT EME AN-CMEAN=
EMEAN=CREAN
WOT SIGNIFICANT EMEAN-CME AN =
125
NOT SIGRIFICAKT EME AN-CME ANS
SIGNIFICANT EME AN-CME AN =
EMEAW ChE An
SIGNIFICANT
EMEAN=CFEAN
NGT SIGNIFICANT EME Ais-CMEAN=

EME AN-CME AN =

EME An-CME AN =

+09 KUM=
+06 NUN=
+00 NUH
+56 NUM
1.07 qUK=
+84 NUN=

-.83
1.60

2.73
-3.67
71.53

33

-el7

230

-.08
247

71.27

-1.17

wuewvew

DEG=
VEG

DEG=
OLG=
DEG=
DEG=
DEG=
OEG=

DEG=
DEG=

DEG=

vELs

4

4

EMEAN=
EMEANS
EMEAN=
EMEAN=
EMEANS
EMEAN=

EMEANS
EMEAN=
EMEAN®
EMEANS
EMEAN=
EMEAN=

EMEANS
EMEAN=
EmEANS
EMEAN=
EMEAN=
EMEAN=

240
6.27
4645
9613
2.52
4.07

21S
4.83
4645
3.47
225
3.27

+25
7.07
445
413
225
3.63

EDEV=
EDEV=
EDEV=
EDEV=
EDEV=
EDEV=

EDEV=
EDEV=

EDEV=
EDEV=

EDEV=
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EDEV=
EDEV=
EDEV=
EOEV=
EDEV=
EDEV=

00
275
75
+68
2.10
23

23
215
oS
215
015
1.62

209
225
225
229
+29
80

CMEAN=
CMEAN=
ChEAN=
CMEAN=
CHEAN=
CMEANS

CHEANS
CMEANS
CMEAN=
CMEAN=
CMEAN=
CMEAN=

CMEAN=
CMEAN=
ChEAN=
CMEAN®
CMEAN=
CHEAN=

26

1.23 COEV=
4.67 COEVE
4.45CDCV=
6.40 CDEV=
6.18 COEV=
5.60 COev=

el SCOEV=
4.50 CDEV=
4.45CDEV=
3.63 COEV=
25C0EV=
2.97 COEV=

33 COEVe
6.60 COEV=
4e45CDEV=
5.40 COEV=

+25C0EV=
4.80 COEV=

57
Sele


APPENDIX B

Results for net uptake for Experiments I and II

A Students t test (P<0.05) was calculated for the null
hypothesis: Ho: uw, = u, where uw, is experimental and Uo is
control for each metal for each combination of Experiment I or II,
at each sample time, with n-2 degrees of freedom shown (Deg).

Also shown are the values of E mean- C mean), the individual
means and standard deviations.

g1


ZN FOULING ORGAN ISMS— Exe. 2

MERN <E-C)> / PPM

soon

2sap

aoap

sop

MEAN PPM

ZN SEWRGE~- EXP. 2 .

Figure 12. Mean net uptake of Zn between experimental and control,
fouling organisms and sewage - Experiment II.

32