50%
!!RH SEDIMENT
FIELD VERIFICATION PROGRAM
(AQUATIC DISPOSAL)
TECHNICAL REPORT D-85-7
USE OF BIOENERGETICS TO INVESTIGATE THE
IMPACT OF DREDGED MATERIAL ON BENTHIC
SPECIES: A LABORATORY STUDY WITH
POLYCHAETES AND BLACK ROCK
HARBOR MATERIAL
by
D. Michael Johns, Ruth Gutjahr-Gobell
Edgerton Research Laboratory
New England Aquarium
Boston, Massachusetts 02110
and
Paul Schauer
Environmental Research Laboratory
US Environmental Protection Agency
Narragansett, Rhode Island 02882
September 1985
Final Report
Approved For Public Release; Distribution Unlimited
Prepared for DEPARTMENT OF THE ARMY
US Army Corps of Engineers
Washington, DC 20314-1000
and US Environmental Protection Agency
Washington, DC 20460
Monitored by Environmental Laboratory
US Army Engineer Waterways Experiment Station
PO Box 631, Vicksburg, Mississippi 39180-0631
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Destroy this report when no longer needed. Do not return
it to the originator.
The findings in this report are not to be construed as an official
Department of the Army position unless so designated
by other authorized documents.
The contents of this report are not to be used for
advertising, publication, or promotional purposes.
Citation of trade names does not constitute an
official endorsement or approval of the use of
such commercial products.
The D-series of reports includes publications of the
Environmental Effects of Dredging Programs:
Dredging Operations Technical Support
Long-Term Effects of Dredging Operations
Interagency Field Verification of Methodologies for
Evaluating Dredged Material Disposal Alternatives
(Field Verification Program)
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SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"Use of Bioenergetics to Investigate the Impact of Dredged Material
on Benthic Species: A Laboratory Study with Polychaetes and Black
Rock Harbor Material"
TO: All Report Recipients
1. This is one in a series of scientific reports documenting the findings of
studies conducted under the Interagency Field Verification of Testing and
Predictive Methodologies for Dredged Material Disposal Alternatives (referred
to as the Field Verification Program or FVP). This program is a comprehensive
evaluation of environmental effects of dredged material disposal under condi-
tions of upland and aquatic disposal and wetland creation.
2. The FVP originated out of the mutual need of both the Corps of Engineers
(Corps) and the Environmental Protection Agency (EPA) to continually improve
the technical basis for carrying out their shared regulatory missions. The
program is an expansion of studies proposed by EPA to the US Army Engineer
Division, New England (NED), in support of its regulatory and dredging mis-
sions related to dredged material disposal into Long Island Sound. Discus-
sions among the Corps' Waterways Experiment Station (WES), NED, and the EPA
Environmental Research Laboratory (ERLN) in Narragansett, RI, made it clear
that a dredging project at Black Rock Harbor in Bridgeport, CT, presented a
unique opportunity for simultaneous evaluation of aquatic disposal, upland
disposal, and wetland creation using the same dredged material. Evaluations
were to be based on technology existing within the two agencies or developed
during the six-year life of the program.
3. The program is generic in nature and will provide techniques and inter-
pretive approaches applicable to evaluation of many dredging and disposal
operations. Consequently, while the studies will provide detailed site-
specific information on disposal of material dredged from Black Rock Harbor,
they will also have great national significance for the Corps and EPA.
4. The FVP is designed to meet both Agencies' needs to document the effects
of disposal under various conditions, provide verification of the predictive
accuracy of evaluative techniques now in use, and provide a basis for deter-
mining the degree to which biological response is correlated with bioaccumula-
tion of key contaminants in the species under study. The latter is an
important aid in interpreting potential biological consequences of bioaccumu-
lation. The program also meets EPA mission needs by providing an opportunity
to document the application of a generic predictive hazard-assessment research
strategy applicable to all wastes disposed in the aquatic environment. There-
fore, the ERLN initiated exposure-assessment studies at the aquatic disposal
site. The Corps-sponsored studies on environmental consequences of aquatic
disposal will provide the effects assessment necessary to complement the EPA-
sponsored exposure assessment, thereby allowing ERLN to develop and apply a
hazard-assessment strategy. While not part of the Corps-funded FVP, the EPA
exposure assessment studies will complement the Corps' work, and together the
Corps and the EPA studies will satisfy the needs of both agencies.
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SUBJECT: Transmittal of Field Verification Program Technical Report Entitled
"Use of Bioenergetics to Investigate the Impact of Dredged Material
on Benthic Species: A Laboratory Study with Polychaetes and Black
Rock Harbor Material"
5. In recognition of the potential national significance, the Office, Chief
of Engineers, approved and funded the studies in January 1982. The work is
managed through the Environmental Laboratory's Environmental Effects of
Dredging Programs at WES. Studies of the effects of upland disposal and
wetland creation are being conducted by WES and studies of aquatic disposal
are being carried out by the ERLN, applying techniques worked out at the
laboratory for evaluating sublethal effects of contaminants on aquatic organ-
isms. These studies are funded by the Corps while salary, support facilities,
etc., are provided by EPA. The EPA funding to support the exposure-assessment
studies followed in 1983; the exposure-assessment studies are managed and
conducted by ERLN.
6. The Corps and EPA are pleased at the opportunity to conduct cooperative
research and believe that the value in practical implementation and improve-
ment of environmental regulations of dredged material disposal will be con-
siderable. The studies conducted under this program are scientific in nature
and will be published in the scientific literature as appropriate and in a
series of Corps technical reports. The EPA will publish findings of the
exposure-assessment studies in the scientific literature and in EPA report
series. The FVP will provide the scientific basis upon which regulatory
recommendations will be made and upon which changes in regulatory implementa-
tion, and perhaps regulations themselves, will be based. However, the docu-
ments produced by the program do not in themselves constitute regulatory
guidance from either agency. Regulatory guidance will be provided under
separate authority after appropriate technical and administrative assessment
of the overall findings of the entire program.
ChoromokosT Jr., Pti.D., P.E.
Director, Research and Development
U. S. Army Corps of Engineers
Bernard D. Goldstein, M.D.
Assistant Administrator for
Research and Development
U. S. Environmental Protection
Agency
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Unclassified
SECURITY CLASSIFICATION OF THIS PAGE (When OMB Entered)
REPORT DOCUMENTATION PAGE
READ INSTRUCTIONS
BEFORE COMPLETING FORM
1. REPORT NUMBER
2. GOVT ACCESSION NO
Technical Report D-85-7
3. RECIPIENT'S CATALOG NUMBER
4. TITLE (and Subtitle)
USE OF BIOENERGETICS TO INVESTIGATE THE IMPACT OF
DREDGED MATERIAL ON BENTHIC SPECIES: A LABORATORY
STUDY WITH POLYCHAETES AND BLACK ROCK HARBOR
MATERIAL
S. TYPE OF REPORT 4 PERIOD COVERED
Final Report
S. PERFORMING ORG. REPORT NUMBER
7. AJTHORf.J
D. Michael Johns, Ruth Gutjahr-Gobell,
Paul Schauer
8. CONTRACT OR GRANT NUMBERf.)
9. PERFORMING ORGANIZATION NAME AND ADDRESS
New England Aquarium Edgerton Research Laboratory,
Central Wharf, Boston, Massachusetts 02110 and
US Environmental Protection Agency, Environmental
Research Laboratory, South Ferry Road,
Narragansett. Rhode Island 02882
\0. PROGRAM ELEMENT, PROJECT, TASK
AREA i WORK UNIT NUMBERS
Field Verification Program
(Aquatic Disposal)
U. CONTROLLING OFFICE NAME AND ADDRESS
DEPARTMENT OF THE ARMY, US Army Corps of Engineers
Washington, DC 20314-1000 and US Environmental
Protection Agency, Washington. DC 20460
t2. REPORT DATE
September 1985
13. NUMBER OF PAGES
14. MONITORING AGENCY NAME & ADDRESS^/ different /ran Controlling Otllce)
US Army Engineer Waterways Experiment Station
Environmental Laboratory
PO Box 631, Vicksburg, Mississippi 39180-0631
IS. SECURITY CLASS, (at thlt report)
Unclassified
IS*. O ECL ASSI FICATtON/DOWNGRADING
SCHEDULE
IS. DISTRIBUTION STATEMENT (ol title Report)
Approved for public release; distribution unlimited.
17. DISTRIBUTION STATEMENT (at the ebeueet entered In Stock 30, II different Inrrt Report)
IS. SUPPLEMENTARY NOTES
Available from National Technical Information Service, 5285 Port Royal Road,
Springfield, Virginia 22161.
19. KEY WORDS (Confirm* on rtreret elde II neceeeery end Identity by block number)
Bioenergetics—Technique (LC)
Dredging—Environmental aspects (LC)
Dredged material (WES)
Polychaeta (LC)
Benthos (LC)
Marine pollution (LC)
Dredging—Connecticut—Black Rock
Harbor (LC)
20. ABSTRACT (Continue ea rente* elite ft mcMMqr end Identify by Mock number)
Both solid phase and particulate phase assays were conducted with two
species of polychaetes to determine the accuracy and reproducibility of con-
ducting bioenergetic studies on poLychaetes exposed to highly contaminated
dredged sediment. The two species tested were Nephtys incisa, an errant
burrowing sediment ingestor, and Neanthes arenaceodentata, a tube-building sur-
face feeder. Exposure to various treatments was for 10 days.
(Continued)
DO
FORM
t JAN 73
1473
EDITION OF I NOV 65 IS OBSOLETE
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SECURITY CLASSIFICATION1 OF THIS PAriC fWi.fi Dele Entered)
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Unclassified
SECURITY CLASSIFICATION OF THIS PA.GEfWh«B D«<« Enl.r.rf)
20. ABSTRACT (Continued).
Results with both species of polychaetes indicate that, with few exceptions,
all of the physiological parameters measured (rates of feeding, growth, repro-
duction, and ammonia excretion) can be made with accuracy. Changes in growth
(determined as dry weight) between treatments, for example, can be measured
following a 10-day exposure period providing that care is taken to adequately
size the individual polychaetes prior to initiation of the experiment.
The bioenergetic endpoints measured in this study were found to be
repeatable. In addition, physiological responses were found to be dose-
dependent. Dosage was based on the relative proportion of reference and Black
Rock Harbor sediment in a particular treatment.
This investigation is the first phase in developing field verified
bioassessment evaluations for the Corps of Engineers and EPA regulatory program
for dredged material disposal. This report is not suitable for regulatory pur-
poses; however, appropriate assessment protocols that are field verified will be
available at the conclusion of this program.
Unclassified
SECURITY CLASSIFICATION OF THIS PAGEfHTiwt D«(» Entered)
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PREFACE
This report describes work performed by the US Environmental Protection
Agency (EPA) Environmental Research Laboratory, Narragansett, R. I. (ERLN),
as part of the Interagency Field Verification of Testing and Predictive
Methodologies for Dredged Material Disposal Alternatives Program (Field
Verification Program (FVP)). This program is sponsored by the Office, Chief
of Engineers, and assigned to the US Army Engineer Waterways Experiment
Station (WES), under the purview of the Environmental Laboratory's (EL)
Environmental Effects of Dredging Programs (EEDP). The OCE Technical Monitors
were Drs. John Hall and William L. Klesch. The objective of this program
agreement Is to verify existing predictive techniques for evaluating the
environmental consequences of dredged material disposal under aquatic, wet-
land, and upland conditions. The aquatic portion of the FVP study is being
conducted by ERLN, with the wetland and upland portion done by WES.
Although not totally inclusive, we would like to thank the following
researchers: Drs. John Gentile, Gerald Pesch, John Scott, Mr. William Nelson,
and Ms. Carole Pesch for their many discussions and criticisms; Mr. Michael
Balboni for the use of and help with their benthic exposure systems;
Ms. Cornelia Meuller for supplying the _N. arenaceodentata juveniles;
Dr. Wayne Davis for use of the 'ant farm'; and Dr. James Heltshe for advice
on statistical procedures. We would especially like to thank Dr. Anthony
Calabrese and Capt. Robert Alix of the National Marine Fisheries Service in
Milford, Conn., for boat time on the R/V Shang Wheeler. This research was
supported by Cooperative Agreement CR809956 between the US Environmental
Protection Agency and the New England Aquarium to Dr. D. Michael Johns.
The EPA Technical Director for the FVP was Dr. John H. Gentile; Technical
-------
Coordinator was Mr. Walter Galloway; and the Project Manager was Mr. Allan
Beck.
The study was conducted under the direct management of Drs. Thomas M.
Dillon and Richard K. Peddicord and under the general management of Dr. C.
Richard Lee, Chief, Contaminant Mobility and Criteria Group; Mr. Donald L.
Robey, Chief, Ecosystem Research and Simulation Division; and Dr. John
Harrison, Chief, EL. The FVP Coordinator was Mr. Robert L. Lazor, and the
EEDP Managers were Mr. Charles C. Calhoun, Jr., and Dr. Robert M. Engler.
COL Tilford C. Creel, CE, and COL Robert C. Lee, CE, were Commanders
and Directors of WES during the conduct of the study. COL Allen F. Grum, USA,
was Director of WES during the preparation and publication of this report.
Mr. Fred R. Brown and Dr. Robert W. Whalin were Technical Directors.
This report should be cited as follows:
Johns, D. M., Gutjahr-Gobell, R., and Schauer, P. 1985.
"Use of Bioenergetics to Investigate the Impact of
Dredged Material on Benthic Species: A Laboratory
Study with Polychaetes and Black Rock Harbor Material,"
Technical Report D-85-7, prepared by US Environmental
Protection Agency, Environmental Research Laboratory,
Narragansett, R. I., for the US Army Engineer Waterways
Experiment Station, Vicksburg, Miss.
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CONTENTS
Page
PREFACE 1
LIST OF FIGURES 4
LIST OF TABLES 5
PART I: INTRODUCTION 7
Background 7
Purpose 8
Scope 8
PART II: MATERIALS AND METHODS 10
Sediment Sources and Exposure Systems 10
Experimental Organisms 17
Physiological Measurements 18
Burrowing Activity 25
Statistical Analysis 27
PART III: RESULTS 30
Sources of Variation 30
Treatment of Data 31
Consumption 32
Production 34
Maintenance Costs 38
Partitioning of Energy Resources 44
Scope for Growth 52
Burrowing Activity 54
PART IV: DISCUSSION 60
PART V: CONCLUSIONS 71
REFERENCES 72
APPENDIX A: SOLID PHASE AND PARTICULATE PHASE DATA SHEETS Al
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LIST OF FIGURES
No.
1 Central Long Island Sound disposal site and South
reference site 11
2 Black Rock Harbor, Connecticut, source of dredged
material 11
3 Sediment dosing system with chilled water bath and
argon gas supply 14
4 Schematic of the suspended distribution and dosing
system used to expose juvenile Nj^ incisa to reference
and BRH sediment 77 16
5 Schematic of respirometers (syringe and stender dish)
used to determine oxygen consumption rates of
N. incisa juveniles 24
6 Schematic of the narrow, glass-walled aquarium used
to determine burrowing activity and maximum burrow
depth 27
7 Net growth efficiency of N_._ incisa juveniles exposed
to various mixtures of reference and BRH sediment for
10 days 45
8 Net growth efficiency of N. incisa juveniles exposed
to various combinations of sediment and suspended
particles for 10 days 48
9 Drawing of burrows created by IJ^ incisa juveniles
exposed to reference and BRH sediments for 10 days.... 54
10 Drawing of burrows created by N. incisa juveniles
exposed to various combinations of reference and
BRH sediment for 10 days 56
11 Drawing of burrows created by N._ arenaceodentata
juveniles exposed to various combinations of
reference and BRH sediment for 10 days 58
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LIST OF TABLES
No. Page
1 Sampling protocol for physiological measurements at the
end of the 10-day exposure period 20
2 Sources of variation in respiration and ammonia
excretion rate of Nj^ incisa juveniles determined during
initial experiments 31
3 Food consumption rates of 11. arenaceodentata juveniles
exposed to various sediment conditions 33
4 Changes in dry weight of N_._ incisa juveniles exposed
to various sediment conditions 35
5 Changes in dry weight of !*._ arenaceodentata juveniles
exposed to various sediment conditions 37
6 Weight-specific respiration rate of N^ incisa juveniles
exposed to various sediment conditions 39
7 Weight-specific respiration rate of N. arenaceodentata
juveniles exposed to various sediment conditions. 40
8 Weight-specific ammonia excretion rate and 0:N ratios of
N. incisa juveniles exposed to various sediment
conditions 42
9 Weight-specific ammonia excretion rate and 0:N ratios
of N. arenaceodentata juveniles exposed to various
sediment conditions 43
10 Cummulative energy values for production and
maintenance costs of 11. incisa juveniles 45
11 Net growth efficiency of N. incisa juveniles exposed to
various sediment conditions 47
12 Cummulative energy values for production and
maintenance costs of IS. arenaceodentata juveniles
exposed to various sediment conditions 50
13 Net growth efficiency of N. arenaceodentata juveniles
exposed to various sediment conditions 51
14 Scope for growth values for Nj^ arenaceodentata
juveniles exposed to various sediments 53
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No. Page
15 Burrow length and maximum burrow depth of N. incisa
juveniles exposed to various sediment conditions 55
16 Burrow length and maximum burrow depth of
N. arenaceodentata juveniles exposed to various
sediment conditions 59
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USE OF BIOENERGETICS TO INVESTIGATE THE IMPACT
OF DREDGED MATERIAL ON BENTHIC SPECIES; A LABORATORY
STUDY WITH POLYCHAETES AND BLACK ROCK HARBOR MATERIAL
PART I: INTRODUCTION
Background
1. Effective short-term biological response measurements
which can adequately detect the effects of environmental concentra-
tions of contaminants have been called for by several international
commissions (International Council for the Exploration of the Sea
1978; Mclntyre and Pearce 1980). In order to be of value the measure-
ments must have some relevance to ecological fitness. In addition,
these relatively short-term laboratory effects tests (usually less
than a month) must have a predictive capability which allows estima-
tion of the degree of ecological change which will take place.
2. An effects measurement technique which may satisfy the
preceding criteria is the determination of biological energy balances
(Edwards 1978; Capuzzo and Lancaster 1981; Johns and Pechenik
1980; Johns and Miller 1982; McKinney 1982) along with its corol-
laries, including scope for growth (Warren and Davis 1967; Bayne
1975). Previous studies using these principles have found a reason-
able correlation between changes in energy balances or scope for
growth and changes in population fitness (Bayne et al. 1979;
Gilfillan 1980). In a series of detailed field studies, for example,
Gilfillan and his co-workers (Gilfillan and Hanson 1975; Gilfillan
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et al. 1976; Gilfillan and Vandermeulen 1978) found a reduced scope
for growth in the bivalve Mya arenaria collected from oil-impacted
sites when compared to individuals from nearby, relatively clean
reference populations. These data were related to and predictive
of eventual changes in population structure observed in the impacted
sites. Changes in population structure that Gilfillan could relate
to the reduced scope for growth included reductions in yearly growth
rate and population density.
Purpose
3. The biological effects portion of the Field Verification
Program (FVP) is being implemented in two phases. The first phase
is to identify, biological test procedures that are responsive to
highly contaminated dredged material. In this phase, the applica-
bility, reproducibility, and repeatability of the biological measure-
ment in the laboratory are to be demonstrated. The second phase will
be to field verify the biological responses observed in the labora-
tory to determine the predictability of the laboratory-derived data.
The purpose of this report is to describe the results of the first
phase in our efforts to apply bioenergetics techniques to two species
of polychaetes.
Scope
4. This paper describes efforts to evaluate the utility of
bioenergetics techniques with the polychaetes Nephtys incisa and
Neanthes arenaceodentata and to determine what effects highly
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contaminated dredged Black Rock Harbor sediment have on these energy
budgets. Nephtys incisa is a dominant infaunal macroinvertebrate
in Long Island Sound benthic communities (Sanders 1956, 1958; Carey
1962). Impact from contaminated dredged material disposal on this
species may change the present community structure. Neanthes
arenaceodentata, on the other hand, is not indigenous to the Long
Island Sound study area. Rather it is being tested as a possible
surrogate for those infaunal polychaete species that may occur at
any disposal site. In disposal sites where indigenous species may be
difficult to collect, or maintain, or study in the laboratory, a
surrogate species offers the opportunity to still determine potential
impacts on that general group of organisms. In proposed disposal
sites, where little scientific information exists on the indigenous
species, the use of a suitable surrogate allows for the assessment
of potential impacts without the time-consuming learning curve
(i.e. methods development, etc.) that would be needed with the
indigenous species.
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PART II: MATERIALS AND METHODS
Sediment Sources and Exposure Systems
5. Reference sediment for these studies was collected from
the FVP South Reference site (40°7.95"N and 72°52.7"W), which is
approximately 700 m south of the southern perimeter of the Central
Long Island Sound disposal site (Figure 1). Reference sediment was
collected with a Smith-Mclntyre grab sampler (0.1 m^) in August and
December 1982 and May 1983 (collections I, II, and III, respectively).
Sediment from each collection was returned to the laboratory, press
sieved (wet) through a 2-mm mesh stainless steel screen, homogenized,
and stored in polypropylene (collection I) or glass (collections II
and III) containers at 4°C until used in experiments. Each container
of material was coded with collection number, date,and jar number.
(See Lake et al. 1984 for complete details.)
6. BRH sediment was collected from 25 locations within the highly
indistrialized Black Rock Harbor (Bridgeport, Conn.; Figure 2) study area
2
with a 0.1-m gravity box corer to a depth of 1.21 m. The contaminated
sediment was homogenized, distributed to barrels, and stored at 4°C. The
contents of each barrel were homogenized, wet sieved through a 1-mm sieve,
distributed to glass jars, and stored at 4°C until used in experi-
ments. Samples of sediment were taken at various points in the
collections, mixing, and distribution procedure for moisture content
and chemical analysis. (See Lake et al. 1984 for details.)
10
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SOUTH REFERENCE
• SITE
Figure 1. Central Long Island Sound disposal site and
South reference site
Figure 2. Black Rock Harbor, Connecticut, source of
dredged material
11
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7. Two exposure regimes were used in this study. One was a
solid phase assay in which the polychaetes were exposed to various
combinations of reference and BRH sediment for 10 days. A 10-day
exposure period was chosen based on data provided in Pesch and
Hoffman (1983). The other exposure regime used was a particulate
phase assay in which worms were exposed to either reference or BRH
particles after being placed in 100 percent reference and 100 percent
BRH, respectively.
8. For the experiments in which the worms were to be exposed
to only the solid phase of the sediments, approximately 400 ml of
the appropriate substrate mixture was placed in a 150- by 75-mm
dish, and then put in a water bath of 20°C (Figure 3). Seawater at
20°C was allowed to flow through the treatment bowls at a rate of
approximately 50 ml/min. Where both reference and BRH sediments
were used in a treatment, the two sediments were combined in a
volume-to-volume ratio and throughly mixed.
9. Implementation of the particulate phase assays required
the consruction of two identical sediment dosing systems to simul-
taneously provide either BRH or reference material as suspended
sediment. The dosing systems (Figure 3) consisted of conical-shaped
slurry reservoirs placed in a chilled fiberglass chamber, a diaphragm
pump, a 4-jj. separatory funnel, and several return loops that
directed the particulate slurry through dosing valves. The slurry
reservoirs (40 cm diam. by 55 cm high) contained 40 I of slurry
composed of 37.7 & of filtered seawater and 2.3 Jl of either BRH
12
-------
or reference sediment. The fiberglass chamber (94 cm by 61 cm by
79 cm high) was maintained between 4°and 10°C using an externally
chilled water source. (The slurry was chilled to minimize microbial
degradation during the test.) Polypropylene pipes (3.8 diam) placed
at the bottom of the reservoir cones were connected to the diaphragm
pumps (16 to 40 fc/min capacity) that had Teflon diaphragms. These
pumps were used to circulate the slurry but minimize abrasion so
that the physical properties and particle sizes of the material
remained as unchanged as possible.
10. The separatory funnel was connected to the pump and
returned to the reservoir by polypropylene pipes. The separatory
funnel served two functions: (a) to ensure that a constant head
pressure was provided by the overflow, and (b) to serve as a connec-
tion for the manifold located 4 cm below the constant head level.
The manifold served to distribute the slurry by directing a portion
of the flow from the funnel (through 6 mm inside diameter polypro-
pylene tubes) through the Teflon dosing valves and back to the
reservoir. At the dosing valves, the slurry was mixed with seawater
to provide the desired concentrations for the toxicity tests.
Argon gas was provided at the rate of 200 ml/min to the reservoir
and separatory funnel to minimize oxidation of the sediment/seawater
slurry. The dosing valves were controlled by a microprocessor.
The microprocessor can be programmed to deliver a pulse with a
duration of 0.1 sec up to continuous pulse delivery and at intervals
from once every second to once every hour.
13
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ARGON
INJECTION
SEPARATORY
FUNNEL
DELIVERY
MANIFOLD
DOSING
VALVE
TO EXPOSURE
SYSTEM
RETURN
MANIFOLD
SLURRY
RESERVOIR
CHILLED
WATER BATH
Figure 3. Sediment dosing system with chilled water bath and
argon gas supply
14
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11. Within the exposure system distribution jar, seawater at
20°C with the appropriate concentration of suspended particles was
allowed to flow through the treatment bowls at a. rate of approximately
35 ml/min (Figure 4). In order to maintain the particles in suspen-
sion as long as possible, a small crystallization dish with a stir
bar was placed in the middle of the exposure dish. For continuity
throughout this report, the scheme used to describe the particulate
phase assay treatments in which there was both suspended and bedded
sediments was: suspended particulate phase/solid phase.
12. In both the solid phase and particulate phase assays 15
worms of approximately the same size were placed in each treatment.
After the 10-day exposure, the worms were sieved out of the mud,
counted, and saved for physiological measurements.
13. During the experiment the worms were offered prawn flakes
(Aquatic Diet Technology, Inc., Brooklyn, N.Y.) as a food source.
Previous research* has indicated that laboratory holdings of K._
incisa grow better when offered prawn flakes than when left without.
It is unclear, however, whether the prawn flakes are used directly
as a food source or whether the flakes act as a substrate for bacte-
rial growth, the bacteria then being utilized as the food source.
Prawn flakes is also the food source used in maintaining laboratory
populations of N. arenaceodentata (Schauer and Pesch, In Preparation).
With this species, the prawn flakes are utilized directly as food.
* Personal communication, Paul Schauer, July 1983, U.S.
Environmental Protection Agency.
15
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DOSING SYSTEM
•Suspended Particles
Spigot-
u
5OOOC
-Distribution Jar
—Stir Bar
EXPOSURE SYSTEM
Stir Bar
\\\
Exposure
"Container
\\\ \\\\\\\\V\\\\\\ \\\\\\\\\\\\ \\\\\vs
Figure 4. Schematic of the suspended distribution and dosing system
used to expose juvenile N^ incisa to reference and BRH
sediment
16
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Experimental Organisms
14. Nephtys incisa were collected from Long Island Sound in
the vicinity of the South Reference site. Bottom samples were
collected using a Smith-Maclntyre grab and were initially sieved on
board the research vessel. Worms which passed through a 2-mm sieve
but were retained on a 0.37-mm sieve were kept for laboratory exper-
imentation and were all juveniles (Carey 1962). The worms were
then placed in a single, unsieved Smith-Maclntyre grab sample for
transport back to the laboratory. In the laboratory, the worms
were resieved with a 0.37-mm mesh using seawater at temperatures
close to collection temperature (+ 2°C).
15. Following this, individuals were visually separated into
relative size classes for experiments. Although somewhat subjective,
careful visual separation leads to a coefficient of variation in
dry weight at the start of an experiment of only 24 percent. Other,
more accurate sizing techniques, such as wet weight determinations
employing a wet-weight-to-dry-weight regression curve, are time-
consuming and, more importantly, may cause physical damage to the
worms from the handling required.
16. All experiments were conducted at 20°C. Laboratory
acclimation for Nephtys incisa collected at 20° + 3°C was a minimum
of 3 days to allow for adjustments to laboratory holding conditions.
For worms collected at temperatures below 16°C, acclimation included
a 1° to 2°C increase in temperature per day until 20°C was attained.
17
-------
17. Neanthes arenaceodentata juveniles used in this study
were from laboratory cultures (original stock from D.J. Reish,
California State University, Long Beach, Calif.)• Worms were cul-
tured in a flow-through system at 20° + 1°C and fed prawn flakes
according to techniques described in Schauer and Pesch (In Prepara-
tion). Worms used in an experiment were from the same hatch and
were of approximately the same age. Refer to summary data sheets
in Appendix A for complete information on the ages of the worms
used in each experiment.
Physiological Measurements
18. Although the fundamental integration of biochemical and
physiological mechanisms is complex and difficult to measure, the
net response can be measured at the whole organism level. Bioener-
getic analysis compares the major anabolic and catabolic processes
that occur and allows for an evaluation of the relative partitioning
of available energy amongst growth and maintenance requirements.
19. The energy budget of the juvenile stage of an organism
can be described by the following formula:
C-P + R + E + F (1)
where
C = total energy consumed
P = amount of energy converted to tissue
R = amount of energy used for maintenance measured via aerobic
respiration
E » amount of energy lost through ammonia excretion
F • amount of energy lost through feces
18
-------
20. Scope for growth (SFG), which is a derivation of the
balanced energy budget formula, is an approach which provides for
an estimation of the potential for growth (Warren and Davis 1967).
In this approach, the amount of energy available for growth (and
reproduction in sexually mature stages) is estimated by the following
relationship:
P = C - F - (R+E) (2)
Since physiological measurements in SFG studies are typically made
over a short period of time (< 24 hr), P in this case is an instan-
taneous measure of growth potential. Scope for growth is an index
by which the current physiological condition of an organism can be
evaluated.
21. The approach taken in this study was to attempt to (a)
measure all parameters in the balanced energy budget equation to
determine reliability of the measurements and ease at which the
measurements can be made and (b) estimate variability in measurement
so that sample size for the experiments can be determined. Following
this, scope for growth values were determined where possible while
at the same time actual growth was being monitored. Table 1 presents
the sampling protocol and the approximate times needed to make the
various physiolgical measurements.
22. For N^ arenaceodentata, values of consumption, produc-
tion, respiration, and excretion were measured directly. Therefore,
scope for growth values as well as balanced energy budgets could be
determined for this species. With N. incisa, however, only values
19
-------
Table 1
Sampling protocol for physiological measurements at the end of
10-day exposure period
Physiological
Measurement
Feeding
Respiration
Excretion
Total Time Required
Feeding
Respiration
Excretion
Total Time Required
Time
Required
hr
Nephtys incisa
2
3
5
Neanthes arenaceodentata
24
4
a*
28
* a » Pespiration and excretion rates determined at same time in syringe
respirometer in a time of 4 hr
20
-------
of production, respiration, and excretion could be measured directly.
Feeding rates, on the other hand, could not be quantitatively meas-
ured with any relability. In failing to quantitatively estimate
food consumption rates, neither scope for growth nor a balanced
energy budget for II. incisa could be derived, although an estimate of
the relative partitioning of energy between production and maintenance
could be made. The measured parameters allowed for the calculation
of a net growth efficiency using the following formula:
Net growth Efficiency = P/(P+R+E) x 100% (3)
23. Net growth efficiency values offer insight into the
degree of integration among physiological processes. It offers a
time course estimate of the cumulative effects a particular
condition has had on an organism. To allow for comparison to N_._
incisa, net growth efficiencies were also calculated for N.
arenaceodentata.
Food consumption and assimilation efficiency
24. Food consumption rates of individual N^ arenaceodentata
were determined over a period of 24 hr using a preweighed amount of
prawn flake as the food source. Following this period, remaining
food was taken from the bowl, rinsed in deionized water, dried to a
constant weight at 60°C, and weighed to the nearest 1 yg on a
Perkin-Elmer AD-2Z Autobalance (Perkin-Elmer Corp., Norwalk, Conn.).
Food consumption rates were taken as the difference in dry weight
of the food initially offered and the amount remaining at the end
21
-------
of 24 hr. In addition to collecting any remaining food particles,
fecal material was also collected, rinsed, dried, and weighed.
25. The efficiency of food assimilation could then be deter-
mined as the difference in energy content between food and fecal
material (energy content of fecal material - energy content of food
source[J/mg]/energy content of food source[J/mg] x 100). The energy
content of both the food and fecal material was determined using a
wet oxidation technique in the presence of an acid-dichrotnate mixture
(Maciolek 1962).
Production
26. In all experiments, juvenile worms were used so that
changes in dry weight reflected only changes in growth (production).
At the beginning of an experiment, a subsample of individuals (10
to 15) was taken to estimate initial worm weight. The worms were
quickly rinsed in deionized water, dried at 60°C for 24 hr, then
weighed on a Perkin-Elmer AD-2Z Autobalance to the nearest ug.
Following the 10-day exposure, 10 to 15 worms were taken from the
experimental conditions to assess their physiological state.
Individual respiration (see paragraphs 27-28) and excretion rates
(see paragraph 31) were determined prior to rinsing, drying,and
weighing. Table 1 presents the time course needed to make the
physiological measurements for both Ni^ incisa and K[._ arenaceodentata.
Changes in growth were computed as being the difference in dry
weight from the beginning to the end of the 10-day exposure period.
After the worms were dried and weighed, energy values for the tissue
22
-------
were determined using a wet oxidation technique in the presence of
an acid-dichromate mixture (Maciolek 1962).
Respiration
27. For N. incisa, routine rates of oxygen consumption were
measured using a 3-cc syringe containing a small amount of surficial
sediment (0.5 to 0.75 cc) as a respirometer (Figure 5). A worm was
placed in each syringe with 1.5 cc of air-equilibrated seawater.
After 1 hr, the oxygen concentration of 1 cc of the seawater was
determined by injecting the sample onto the face of a Radiometer
oxygen probe (Radiometer, Copenhagen, Denmark) fitted with a Radio-
meter thermostated sampling cell. The syringe was then refilled to
1.5 cc, and the new oxygen concentration recalculated. This proce-
dure was repeated once per worm. Three control syringe respirometers
containing 0.5 to 0.75 cc of sediment were included in each experiment,
28. For N. arenaceodentata, routine rates of oxygen consump-
tion were measured using a 10-cc syringe (filled to 8 cc) containing
no sediment. Rather, worms were offered 5-mm (outside diameter)
glass tubes inside which they would begin to form a mucus tube.
This glass tube could then be handled without any apparent effect
on the worms. After 2 hr, the oxygen concentration of the seawater
was determined by injecting a 1-cc sample onto the oxygen probe.
With this species, the syringe was not refilled as the remaining
seawater (7 cc) was used to determine ammonia excretion rates.
29. Stender dishes containing approximately 25 ml of sedi-
ment were used to test the validity of oxygen consumption rates of
23
-------
N. incisa obtained from the syringe respirometers. A 35- by 50-mm
stender dish with a hole drilled in the side was fitted with a
Radiometer oxygen probe (Figure 5). Declining oxygen tension in
the dish was monitored using a strip chart recorder. The water
within the dish was stirred using a magnetic stir bar held to the
top of the respirometer by a water-driven magnetic stirrer. Control
runs were made with the sediment alone to determine the amount of
oxygen depletion due solely to the biological oxygen demand of the
sediment. The result of this test indicated that there was no
significant difference in measuring respiration rate in syringes
than in the larger sediment-filled stender dishes (syringe: 1.59 +
0.61 yl 02/mg/hr; stender dish: 1.33 + 0.54 yl 02/mg/hr). This
experiment was repeated 6 times, with one stender dish respirometer
and five syringe respirometers per experimental run.
RESPIROMETERS
TO 0
METER
RECORDER
CHAMBER
SYRINGE
MAGNETIC STIRRER
— STIR BAR
SEDIMENT
Figure 5. Schematic of respirometers (syringe and stender dish)
used to determine oxygen consumption rates of N. incisa
juveniles
24
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30. In all experiments oxygen consumption rates are reported
as weight-specific rates. When reported, the energy expended during
routine metabolism was considered to be the average oxygen consumption
rate (microliters per hour) during the experimental period times
the energy equivalent for oxygen (4.80 cal/ml; Elliot and Davison
1975).
Excretion
31. Following their use in the respiration tests, individual
NL_ incisa were placed in 6 ml of filtered seawater without sediment
for approximately 3 hr to determine ammonia excretion rates. Follow-
ing this period of time, the dry weight of each worm was taken.
For N^ arenaceodentata, the water for ammonia determination was
taken directly from the syringe respirometers.
32. Ammonia concentrations were determined according to the
technique of Bower and Holm-Hansen (1980). Calories lost per unit
time as excreted ammonia were calculated by multiplying the excretion
rate of an individual worm (micrograms of NH4N per hour) during the
experimental period by the heat of formation of ammonia (0.62 cal/mg;
Elliot and Davison 1975). Atomic ratios of oxygen consumed to
ammonia excreted (0:N ratios) were calculated following the method
of Corner and Cowey (1968).
Burrowing Activity
33. Quantitative estimates could not be made of the feeding
rate for Nephtys incisa. Since this species is thought to ingest
25
-------
sediment in its search for food (i.e., burrow), it was felt that
making quantitative measurements on burrowing activity could provide
a qualitative estimate of feeding rates. Burrowing activity of N_._
incisa juveniles in the solid phase assay treatments was estimated
using narrow, glass-walled aquaria. The aquaria were 14 cm long by
1 cm wide and 15 cm high (Figure 6; Davis 1979). Individual aquaria
were filled with homogenized mixtures of the appropriate sediment
and allowed to equilibrate in a flowing seawater bath. Following
this equilibration period (1 to 2 hr), two juvenile worms were
randomly placed on the sediment surface. At the end of the 10-day
exposure period, plastic sheets were then laid against the glass
walls of the aquarium and the visible burrows traced. Total burrow
lengths were calculated from these tracings using a Numonics 1224
electronic digitizer (Numonics Corp., Lansdale, N.J.). In the
particulate phase assays, a tracing was made of those burrows (for
both N_^ incisa and N_._ arenaceodentata) visible on the side of the
exposure dish.
34. As the narrow aquaria restrict worm movements to essen-
tially two dimensions, data collected from these aquaria do not
necessarily reflect the normal patterns and depths that might be
expected in the field from the sizes of worms used. They do, however,
provide a relative index of the worm activity in the various treatment
conditions.
26
-------
16cm
14cm
Figure 6. Schematic of the narrow, glass-walled aquarium used to
determine burrowing activity and maximum burrow depth
Statistical Analysis
35. In order to satisfy the objectives of this study, it was
important to establish the variation expected from an individual
worm when making physiological measurements as well as the degree
of variation expected between individuals. To establish this,
repeated rate determinations (respiration or excretion rate) were
made on a series of individual N_._ incisa.
36. Sources of variation in physiological rate measurements
can include: (a) temporal changes in the rate within an organism,
(b) differences in rates between organisms, and (c) the effects of
a particular treatment on the physiological rate. Of prime importance
27
-------
in effects studies is an ability to detect significant changes in
the measured parameter (i.e., physiological rate) due to the treat-
ment. Within-organism and between-organism variation, therefore,
must be quantified so that appropriate sample sizes needed to detect
a treatment effect can be determined. The between-worm and within-
worm variation for the physiological measure was then determined
using the VARACOMP procedure (Goodnight 1979).
37. Based on these variations, sample size was then calcu-
lated using a general sample size formula (Snedecor and Cochran
1980). The following sample size determination formula was used to
determine the minimum sample size needed:
n = C x (2S)/D (4)
where
n = sample size
C » constant which is a function of a type I and type II error
S = estimate of variability from the pilot study
D = percentage deviation from the mean that is to be
detected (Snedecor and Cochran 1980)
38. Where appropriate within an assay (solid phase or par-
ticulate phase), data from a particular treatment from each repeated
experiment were combined for analysis. The decision to combine
these data was based on the fact that within each assay there were
no significant differences in worm size or respiration or excretion
rates at the start of the experiment. It can therefore be concluded
that there were no major physiological differences in the worms
from each experiment at the beginning of the 10-day assay.
28
-------
39. A one-way analysis of variance was computed to determine
the effects of the various treatments on the physiolgical function
of both polychaetes (Snedecor and Cochran 1980). If significant
differences (at P = 0.05) were found, a Tukey and Kramer pairwise
comparison was used to determine where the differences existed
(Snedecor and Cochran 1980). Data on burrow length and maximum
burrow depth were rank transformed prior to one-way analysis of
variance (Conover and Iman 1981).
29
-------
PART III: RESULTS
Sources of Variation
Respiration rate
40. In this study, the greatest variance found in the
respiration rates of N. incisa juveniles was the between-worm compo-
nent (Table 2). Assuming that a 50 percent change in respiration
rate is biologically significant and is the percent change at which
treatment effects should be detectable, a sample size of 10 was
found to be sufficient based on a level of confidence set at 90
percent, using the between-worm variation as the estimate of varia-
bility. If a 25 percent change in respiration rate had been chosen
as the desired level, a sample size of 22 would be needed to detect
significant differences.
Excretion rate
41. Using the approach outlined in paragraph 36, sources of
variation were determined for ammonia excretion rates in N. incisa.
As with respiration rates, the between-worm component was the greatest
source of variation (Table 2). Because of the low variation found
in the between- and within-worm variation for ammonia excretion
rates, only 3 worms would be needed to detect significant differences
at the 90 percent confidence level. However, ten worms were used
in order to provide a complementary set of measurements to the
respiration rate determinations.
30
-------
Table 2
Sources of variation in respiration and ammonia excretion rate
of N. "incisa juveniles determined during initial experiments
Respiration Rate* N** Excretion Rate* H**
Within Worm Variance
(Repeated measurements)
Between Worm Variance
0.166
0.340
25 0.00000391 25
25 0.00000624 25
* Variance.
** N= Number of Determinations.
Treatment of Data
Nephtys incisa
42. Within an assay, no significant differences were found
in worm size and respiration and excretion rates of N. incisa at the
start of the experiment. Therefore,data from each individual exper-
iment (within an assay) were combined for analysis. Summary data
from each individual experiment can be found in Appendix A.
Neanthes arenaceodentata
43. Unlike N. incisa, there were significant differences in
worm size at the beginning of each experiment within an assay. For
this reason, data from the various experiments within the particular
assay were analyzed separately.
31
-------
Consumption
44. Food consumption rates of N. arenaceodentata juveniles
exposed to 100-percent reference sediment for 10 days were signif-
icantly higher than those exposed to 100-percent BRH sediment (Table
3). Feeding rates of those worms exposed to the 50:50 mixture of
reference and BRH sediments were not significantly different from
those of the control group although they were significantly higher
than those worms exposed to 100-percent BRH.
45. Juvenile N. arenaceodentata exposed in the particulate
phase assays exhibited similar food ingestion patterns to those
found in the solid phase assays. In experiment I, consumption
rates of worms from the reference/reference (particulate phase/solid
phase) and reference/BRH treatments were significantly higher than
those rates for worms exposed in either the BRH/reference or BRH/BRH
regimes (Table 3). In experiment II, there were significantly
higher food consumption rates in juveniles from the reference/refer-
ence and BRH/reference treatments compared to those from the BRH/BRH
exposure (Table 3). There was no difference, however, between refer-
ence/BRH and BRH/BRH ingestion rates as well as no significant
difference between reference/BRH and both reference/reference and
BRH/reference treatments (Table 3).
32
-------
Table 3
Food consumption rates of N. arenaceodentata juveniles
exposed to various sediment conditions
Experiment Treatment
I 100% REF*
50% REF/ 50% BRH
100% BRH
I REF/REF
REF/ BRH
BRH/ REF
BRH/ BRH
II REF/REF
REF/BRH
BRH/ REF
BRH/ BRH
Consumption Rates**
mg/24 hr
Solid Phase Assay
4.49 + 0.68
4.34 + 0.72
3.52 + 0.85
Particulate Phase Assay
4.49 + 0.96
4.63 + 1.58
3.31 + 1.23
3.38 + 1.23
6.65 + 0.92
6.35 + 1.14
6.62 + 1.36
5.54 + 1.00
Nt Gptt
5 A
5 A
5 B
10 A
10 A
10 B
10 B
10 A
10 A,B
10 A
10 B
* Ref =• Reference Sediment
** Means + S.D.
t N - Number of Determinations
tt Gp - Grouping Letter. Means having the same Gp are not
significantly different at p - 0.05.
33
-------
Production
Nephtys incisa
46. Exposure to 100-percent BRH sediment for 10 days had a
significant effect on the growth of juvenile Nephtys incisa (Table
4). Worms maintained in 100-percent reference and 50-percent refer-
ence/50-percent BRH sediment conditions gained an average dry weight
of 24 and 9 percent, respectively, during the 10-day exposure,
while those worms in the 100-percent BRH treatment lost an average
of 16 percent of their dry weight during the same period.
47. Juvenile N^ incisa exposed in the particulate phase
assays exhibited similar changes in dry weight (Table 4). In both
experiments conducted with these conditions, worms from the reference/
reference treatments (particulate phase/solid phase) were significantly
heavier at the end of the 10-day exposure than those exposed to
BRH/BRH. Worms from the BRH/reference treatments were not signifi-
cantly different from the reference/reference worms.
34
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Table 4
Changes in dry weight of N. incisa juveniles exposed to
various sediment conditions
Change In
Weight
Gpt mg %
Dry Weight**
Initial Final
Treatment mg mg
Solid Phase Assay
100% REF* 1.411+0.246 1.743+0.204 A +.33 ; +24
50% REF/50% BRH 1.411 + 0.246 1.543 + 0.389 A +.13 ; + 9
100% BRH 1.411+0.246 1.189+0.105 B -.22; -16
Particulate Phase Assay
REF/REF
REF /BRH
BRH/REF
BRH/BRH
3
3
3
3
.277
.277
.277
.277
+
+
+
+
0.762
0.762
0.762
0.762
4
3
3
3
.173
.742
.850
.228
+ 0
+ 0
+ 0
+ 0
.535
.755
.723
.578
A
A
A
B
+.95
+ .47
+.57
-.05
; +29
; +14
; +18
; -2
* Ref - Reference Sediment
** Means +1 S.D.
t Gp - Grouping letter. Means having the same Gp are not significantly
different at p - 0.05
35
-------
Neanthes arenaceodentata
48. Exposure to 100-percent BRH sediment did not significantly
alter growth of N_. arenaceodentata during the 10-rday exposure period
(Table 5) although the trend was for the mean dry weight of the 100-
percent BRH-exposed worms to be lower than for the worms exposed in
the reference sediment.
49. For experiment I of the particulate phase assay, there
was no significant difference in worm dry weights at the end of a
10-day exposure to the various sediment combinations. In experiment
II, however, growth of those N. arenaceodentata Juveniles exposed
to the BRH/BRH conditions was significantly lower than exhibited by
worms from the other conditions (Table 5).
36
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Table 5
Changes in dry weight of N. arenaceodentanta juveniles exposed
to various sediment
Experiment Treatment
Dry V
Initial
mg
Solid Pt)
I 100% REF*
50% REF/50% BRH
100% BRH
4.
4.
4.
16 +
16 +
16 +
1
1
1
.73
.73
.73
Particulate
I REF/REF
REF /BRH
BRH/REF
BRH/BRH
II REF/REF
REF /BRH
BRH/REF
BRH/BRH
3.
3.
3.
3.
4.
4.
4.
4.
52 +
52 +
52 +
52 +
43 +
43 +
43 +
43 +
2
2
2
2
0
0
0
0
.09
.09
.09
.09
.92
.92
.92
.92
conditions
/eight**
Final
mg Gp*
iase Assay
6.44
5.03
5.16
Phase
5.41
5.98
5.87
5.35
6.51
6.27
6.26
4.64
+ 1
+ 1
+ 1
.69
.21
.08
A
A
A
Change In
Weight
mg %
+2
+0
+1
.28
.87
.00
+55
+21
+24
Assay
+ 1
+ 1
+ 1
+ 1
+ 1
+ 1
+ 1
+ 0
.80
.47
.95
.58
.91
.93
.53
.88
A
A
A
A
A
A
A
B
+1
+2
+2
+1
+2
+1
+1
+0
.89
.46
.35
.83
.08
.84
.83
.21
+54
+70
+67
+52
+47
+42
+41
+5
* REF - Reference Sediment.
** Means +1 S.D.
t Gp - Grouping letter. Means having the same Gp are not
significantly different at p - 0.05.
37
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Maintenance Costs
Respiratory expenditures
50. Nephtys incisa. Weight-specific respiration rates of
N^ incisa juveniles exposed to 100-percent reference sediment were
significantly higher than those of juvenile worms exposed to 100-
percent BRH sediment (Table 6). Worms from the 50-percent reference/
50-percent BRH treatment exhibited oxygen consumption rates inter-
mediate to those from the 100-percent reference and 100-percent BRH
conditions.
51. A similar pattern was found for those worms from the
particulate phase experiments. In this assay, N. incisa juveniles
from the reference/reference treatment had significantly higher
respiration rates than those worms from the BRH/BRH exposure (Table
6). Respiration rates of worms from the BRH/reference treatment
were also significantly higher than those of the BRH/BRH-treated
worms but were not statistically different from the reference/refer-
ence individuals. Worms from the reference/BRH treatment exhibited
oxygen consumption rates similar to those found in the BRH/BRH-
treated worms (Table 6).
52. Neanthes arenaceodentata. No significant differences in
weight-specific respiration rate were found in N. arenaceodentata
exposed to the various sediment treatments (Table 7).
38
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Table 6
Weight-specific respiration rate of N. incisa juveniles
exposed to various sediment conditions
Treatment
Respiration Rate**
yd 07/mg/hr
N1
Gp
TT
Solid Phase Assay
100% REF*
50% REF/ 50% BRH
100% BRH
REF/REF
REF/BRH
BRH/REF
BRH/BRH
1.65 + 0.77
1.36 + 0.63
1.10 + 0.73
Particulate
0.75 + 0.20
0.62 + 0.25
0.86 + 0.25
0.66 + 0.28
62 A
56 A,B
44 B
Phase Assay
42 A
42 B
42 A
44 B
* REF » Reference Sediment.
** Means + 1 S.D.
tN= Number of determinations.
tt Gp - Grouping Letter. Means having the same Gp are not
significantly different.
39
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Table 7
Weight-specific respiration rate of N. arenaceodentata juveniles
exposed
Experiment Treatment
I 100% REF*
50% REF/ 50%
100% BRH
I REF/REF
REF/ BRH
BRH/REF
BRH/ BRH
II REF/REF
REF/ BRH
BRH/REF
BRH/BRH
to various sediment conditions
Respiration Rate**
11 «. 0,/mg/hr NT
Solid Phase Assay
1.23+0.17 4
BRH 1.18+0.27 5
0.94+0.19 7
Particulate Phase Assay
2.30+0.61 10
2.23 + 0.65 10
2.05 + 0.70 10
2.57 + 0.74 10
1.74 + 0.41 10
1.93+0.50 10
1.81 + 0.31 10
2.16 + 0.37 9
GP"
A
A
A
A
A
A
A
A
A
A
A
* REF - Reference Sediment.
** Means + 1 S.D.
t N - Number of Determinatons.
ft Gp » Grouping letter. Means having the same Gp are not
significantly different.
40
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Ammonia excretion rates and 0;N ratios
53. Nephtys incisa. As with weight-specific respiration
rates, weight-specific ammonia excretion rates of N. incisa juveniles
exposed to 100-percent BRH and BRH/BRH conditions were significantly
lower than those rates of worms exposed to 100-percent reference
and reference/reference (Tables 8). The other treatments were
intermediate between these two values.
54. No significant differences were found in 0:N ratios
between the various treatments in either the solid phase or the
particulate phase assay (Table 8). In all cases, 0:N ratios were
relatively high (greater than 50) indicating that lipids and carbo-
hydrates are being used as part of the substrate for energy produc-
tion.
55. Neanthes arenaceodentata. For two of the three experi-
ments conducted with N_._ arenaceodentata (solid phase experiment and
experiment I, particulate phase), no significant differences in
weight-specific ammonia excretion rates were found (Table 9). In
the other particulate phase experiment, however, a significant
increase in ammonia excretion rate was detected in those worms
exposed to either the BRH/BRH or BRH/reference treatment.
56. As with N_-_ incisa no significant differences were found
in 0:N ratios between the various experiments in either assay. The
0:N ratios in all treatments ranged between 84 and 229 (Table 9).
41
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Table 8
Weight-specific ammonia excretion rate and OtN ratios of N. incisa
Treatment
100% REF*
50% REF/ 50%
100% BRH
REF/ REF
REF/ BRH
BRH/ REF
BRH/ BRH
juveniles exposed to various sediment
Excretion Rate**
v$ NH4N/mg/hr Nt Gptt
Solid Phase Assay
0.011 + 0.003 62 A
BRH 0.010 + 0.002 56 A,B
0.008 + 0.004 44 B
Particulate Phase Assay
0.012 + 0.003 42 A
0.010 + 0.002 42 B
0.013 + 0.004 42 A
0.010 + 0.004 44 B
conditions
0:N
Ratio**
147 + 69
121 + 56
98 + 60
67 + 18
55 + 22
77 + 22
59 + 25
A
A
A
A
A
A
A
* REF - Reference Sediment.
** Means + 1 S.D.
t N « Number of determinations, all three experiments combined.
Tt Gp - Grouping letter. Means with the same Gp are not
significantly different.
42
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Table 9
Weight-specific ammonia excretion rate and OtN ratios of N. arenaceodentata
juveniles
exposed to various sediment conditions
Experiment Treatment
I 100% REF*
50% REF/ 50%
100% BRH
I REF/ REF
REF/ BRH
BRH/ REF
BRH/ BRH
II REF/REF
REF/BRH
BRH/REF
BRH/BRH
Excertion Rate**
ug NH4N/mg/hr Nt
Solid Phase Assay
0.0045 + 0.001 4
BRH 0.0034 + 0.001 5
0.0050 + 0.001 7
Particulate Phase Assay
0.010 + 0.007 10
0.011 + 0.004 9
0.015 + 0.007 8
0.013 + 0.007 7
0.008 + 0.002 9
0.008 + 0.004 10
0.013 + 0.003 10
0.014 + 0.004 9
0:N
Gptt Ratio** Gptt
A 110 + 21 A
A 105 + 17 A
A 84 + 25 A
A 205 + 55 A
A 199 + 58 A
A 183 + 62 A
A 229 + 66 A
B 155 + 37 A
B 172 + 45 A
A 161 + 32 A
A 193 + 42 A
* REF • Reference sediment.
** Means + 1 S.D.
t N - Number of determinations.
tt Gp - Grouping letter. Means with the same Gp are not
significantly different.
43
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Partitioning of Energy Resouces
Nephtys incisa
57. The preceding physiological data were used to calculate
the efficiency at which available energy (P+R+E) was partitioned
between growth and maintenance (Respiratory energy expenditure and
Excretory energy losses). In both the solid phase and particulate
phase experiments, maintenance costs were higher in the reference
treatments (100-percent reference; reference/reference) than they
were in the Black Rock Harbor treatments (100-percent BRH; BRH/BRH)
(Table 10). Estimated maintenance costs for the worms from the
other treatments were generally intermediate between these two
extremes.
58. Energy losses through ammonia excretion were not impor-
tant when compared to other energy costs involved in maintenance
(i.e., respiratory expenditures; Table 10). However small, there
were significant differences in energy lost when the excretion
rates of worms maintained in 100-percent reference and reference/
reference were compared to 100-percent BRH and BRH/BRH.
-------
Table 10
Cummulative energy values for production and maintenance costs
of N. incisa juveniles
Respiratory Excretory
Growth** Energy Expenditure** Energy Loss**
Treatment J^ Gptt _jt Gptt jt Gptt
Solid Phase Assay
100% REF* 4.85 + 0.97 A 12.6 + 3.8 A 0.035 + 0.009 A
50% REF/50% BRH 2.01 + 0.48 B 8.9 + 3.6 A,B 0.029 + 0.008 A,B
100% BRH -2.30+0.36 C 5.6+2.1 B 0.020+0.006 B
Particulate Phase Assay
REF/REF 9.77 + 1.86 A 14.1 + 4.7 A 0.077 + 0.018 A
REF/BRH 4.62 + 0.92 B 11.2 + 3.0 B 0.064 + 0.018 B
BRH/REF 6.44 + 1.42 A 15.8 + 5.0 A 0.084 + 0.027 A
BRH/BRH -1.73+0.52 C 9.8+3.1 B 0.054+0.012 B
* REF » Reference Sediment.
** Means + 1 S.D.
t J » Joules.
tt Gp » Grouping letter. Means having the same Gp are not
significantly different at p - 0.05.
45
-------
59. Comparison of net growth efficiencies of N. incisa
juveniles from the different treatments showed that worms maintained
in 100-percent reference sediment were most efficient (+27 percent)
at converting available energy to new tissue (Table 11; Figure 7).
Worms maintained in 100-percent BRH were the least efficient (-24
percent) and in fact lost weight during the exposure period. In
the 50-percent reference/50-percent BRH treatment, worms converted
energy to tissue with an efficiency of +17 percent, which was not
statistically different from the efficiency for worms maintained in
the 100-percent reference condition.
60-i
40-
I 20-
UJ
S+
u_
LU
0-
o
(E
UJ
20-
40-
60J
0% 50%
% BRH SEDIMENT
100%
Figure 7.
Net growth efficiency of N. incisa juveniles exposed
to various mixtures of reference and BRH sediment for
10 days.
-------
Table 11
Net Growth Efficiency of N. incisa juveniles exposed
to various sediment conditions
Net Growth
Treatment Efficiency**
Solid Phase Assay
100% REF* +27 + 9 A
50% REF/ 50% BRH +17 + 9 A
100% BRH -24 +30 B
Particulate Phase Assay
REF/REF +39 +10 A
REF/BRH +32 +16 A
BRH/REF +27 +7 A
BRH/ BRH -28 +40 B
* REF - Reference Sediment.
** Means +1 S.D.
t Gp - Grouping letter. Means having the same Gp are not
significantly different at p - 0.05.
47
-------
60. A similar pattern of net growth efficiency was found in
the particulate phase assays. N. incisa juveniles maintained under
reference/reference conditions exhibited the highest net growth
efficiency (+39 percent) while those in the BRH/BRH treatment had a
negative net growth efficiency (-28 percent) (Table 11; Figure 8).
Net growth efficiencies of worms kept in reference/BRH and BRH/refer-
ence exposures were +32 and +27 percent, respectively. These values
were not significantly different from those of the worms in the
reference/reference conditions.
60-
o
g 20
o
-
lt.
UJ
0-
O
cc.
UJ
40-
60J
con/con con/BRH BRH/con BRH/BRH
SEDIMENT COMBINATION
(particulate/ sediment)
Figure 8. Net growth efficiency of N^ incisa juveniles exposed to
various combinations of sediment and suspended particles
for 10 days
48
-------
Neanthes arenaceodentata
61. The appropriate data were also used to calculate cummula-
tive energy values and net growth efficiency for N. arenaceodentata.
As can be seen in Table 12, a great amount of variance was connected
with estimating tissue production during the 10-day experimental
period. As with Nephtys incisa, energy losses through ammonia
excretion were not important when compared to energy costs involved
in other maintenance processes (respiratory energy expenditure;
Table 12).
62. The large variation in production in turn leads to a
large variation in calculated net growth efficiencies (Table 13).
The mean net growth efficiencies of the worms maintained in the
reference sediment, however, are in some agreement with those found
for N. incisa. In experiment II of the particulate phase assay,
the pattern of net growth efficiency is similar to that of N^
incisa. In this experiment, the net growth efficiencies of worms
exposed to the BRH/BRH treatment was significantly lower than it
was for worms in the other treatments.
49
-------
Table 12
Cummulative energy values for production and maintenance
costs of N. arenaceodentata juveniles exposed to various
sediment conditions
Respiratory Excretory
Growth** Energy Expenditure** Energy Loss**
Experiment Treatment .P Gp" jt Gptt jt Gp^
Solid Phase Assay
I 100% REF* 22+20 A 38+9 A 0.057 + 0.016 A
50% REF/50% BRH 14+19 A 30 + 11 A,B 0.028 + 0.001 B
100% BRH 13+14 A 23+3 B 0.041 + 0.008 A
Particulate Phase Assay
I REF/REF
REF /BRH
BRH/REF
BRH/BRH
II REF/REF
REF/BRH
BRH/REF
BRH/BRH
39
45
49
37
36
38
38
3
+ 38
+ 27
+ 40
+ 32
+ 33
+ 39
+ 31
± 14
A
A
A
A
A
A
A
B
57 +
61 +
54 +
62 +
52 +
56 +
53 +
47 +
13
8
13
7
6
10
6
9
A
A
A
A
A
A
A
A
0
0
0
0
0
0
0
0
.081
.108
.140
.126
.080
.083
.140
.113
+
+
+
+
+
+
+
+
0.040
0.027
0.052
.069
0.021
0.028
0.030
0.029
A
A
A
A
A
A
A
A
* REF - Reference Sediment.
** Means +1 S.D.
t J - Joules.
tt Gp - Grouping letter. Means with the same Gp are not significantly
different.
50
-------
Table 13
Net growth efficiency of N. arenaceodentata juveniles exposed
to various sediment conditions
Net Growth
Experiment Treatment Efficiency** Gpt
Solid Phase Assay
100% REF* +29 +34 A
50Z REP/50Z BRH +18+42 A
100% BRH +27 + 32 A
Particulate Phase Assay
I
II
REF/REF
REF/BRH
BRH/REF
BRH/BRH
REF/REF
REF/BRH
BRH/REF
BRH/BRH
+30 + 27
+39 + 16
+41 + 20
+31 + 23
+35 + 19
+34 + 24
+35 + 20
+ 3 + 23
A
A
A
A
A
A
A
B
* REF « Reference Sediment.
** Means + 1 S.D.
t Gp • Grouping letter. Means having the same Gp are not
significantly different at p - 0.05.
51
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Scope for Growth
63. In order to calculate scope for growth (SFG) values, the
energy content of the food offered to N. arenaceodentata had to be
determined. In addition, the effeciency at which ingested food was
assimilated had to be estimated. The prawn flake used as a food
source in these experiments was found to have an energy value of
18.53 + .471 J/mg while the assimilation efficiency was estimated to
be between 20 and 27 percent.
64. In all experiments, the SFG following a 10-day exposure
to BRH sediment (100-percent BRH or BRH/BRH treatments) was signifi-
cantly reduced (Table 14). The SFG values from these treatments
represent a 27- to 65-percent reduction in the growth potential
compared to SFG values for worms from the reference sediment condi-
tions (100-percent reference and reference/reference).
52
-------
Table 14
Scope for growth for N. arenaceodentata juveniles
exposed to various sediments
Scope for Growth**
Experiment Treatment J/Day Gpt
Solid Phase Assay
100% REF* 12.1 + 2.5 A
50% REF/50% BRH 10.8 + 1.4 A,B
100% BRH 8.8 + 2.5 B
Particulate Phase Assay
II
REF/REF
REF/BRH
BRH/REF
BRH/BRH
I REF/REF
REF/BRH
BRH/REF
BRH/BRH
7.2 + 2.1
6.8 + 3.1
2.6 + 2.1
2.5 + 2.9
17.8 + 3.1
16.8 + 4.0
17.6 + 4.2
10.1 + 2.9
A
A
B
B
A
A
A
B
* REF - Reference Sediment.
** Means + 1 S.D.
t Gp - Grouping letter. Means having the same Gp are not
significantly different at p - 0.05.
53
-------
Burrowing Activity
Nephtys incisa
65. Both burrow length and maximum depth of burrowing in the
100-percent reference treatment were significantly different from
similar measurements taken from the 100-percent BRH exposure (Table
15). The total length of visible burrows was approximately three
times greater in the 100-percent reference condition than it was in
100-percent BRH (Figure 9). The depth to which the worms burrowed
was three times greater in 100-percent reference sediment (30.4 mm)
when compared to the maximum burrow depth in 100-percent BRH (9.5
mm) .
SOUTH REFERENCE SEDIMENT
Burrow Length = 42.2 ±9.9 cm
Maximum Burrow Depth = 30.417.3 mm
BLACK ROCK HARBOR SEDIMENT
Figure 9.
Burrow Length = 14.2 + 4.7 cm
Maximum Burrow Depth = 9.5 ±2.3 mm
Drawing of burrows created by N. incisa juveniles exposed
to reference and BRH sediments for 10 days
54
-------
Table 15
Burrow length and maximum burrow depth of N. inclsa juveniles exposed
to various sediment conditions
Treatment
Burrow Length** Percent in Maximum Burrow Depth**
cm Gptt Sediment mm Gptt
100% REF*
50% REF/ 50% BRH
100% BRH
REF/REF
REF/ BRH
BRH/ REF
BRH/ BRH
Solid Phase Assay
42.2 + 9.9 A - 30.4 + 7.3
N.D.t - N.D.
14.2 + 4.7 B - 9.5 + 2.3
Particulate Phase Assay
43.1 + 8.9 A 58.7 29.3 + 2.0
21.8 + 1.1 B 40.8 17.0 + 2.4
36.9+6.2 A 77.0 31.2+6.1
9.3 + 0.8 C 31.3 16.8 + 2.5
A
B
A
B
A
B
* REF - Reference Sediment.
** Means +1 S.D.
t N.D. - No data.
tt Gp - Grouping letter. Means with the same Gp are not significantly
different at p - 0.05.
55
-------
66. Results of measurements of burrowing activity in the
particulate phase experiments were similar to the preceding results,
although somewhat complicated by the fact that suspended particles
settled out during the course of the experiment to form a layer of
sediment over the bedded sediment. Nephtys incisa from the reference/
reference condition produced significantly more burrows and burrowed
to a significantly greater depth than did those worms maintained in
BRH/BRH (Table 15; Figure 10).
PARTICULATE/SOLID PHASE
control/control
burrow length =43.1 ±8.9
BRH/control
burrow length = 36.9 ±6.2
control/BRH
burrow length = 21.8± I.I
BRH/BRH
burrow length = 9.310.8
Figure 10. Drawing of burrows created by N_._ incisa juveniles exposed
to various combinations of reference and BRH sediment
for 10 days
56
-------
67. Burrowing activity in the other two particulate phase
treatments depended on whether the reference sediment was in the
upper particulate layer or was present as the original, bedded
sediment. In the BRH/reference treatment, most of the burrowing
activity was concentrated in the original sediment (77 percent of
activity) and the depth of burrowing was not significantly different
from that found in the reference/reference treatment (Table 15;
Figure 10). In the reference/BRH exposure, on the other hand, most
of the burrowing activity (59 percent) was found in the particulate
layer which formed during the 10-day exposure. Maximum burrow
depth in this treatment was only 17 mm, which is very similar to
the maximum burrow depth found in the BRH/BRH treatment.
Neanthes arenaceodentata
68. Neanthes arenaceodentata juveniles exhibited significantly
greater burrowing activity (both burrow length and maximum depth of
burrowing) in those treatments that had reference sediment as the
bedded sediment than in those worms which had BRH as the bedded
sediment (Table 16; Figure 11).
69. In all treatments, more than half of the burrowing activity
occurred in the upper particulate layer which formed during the
10-day exposure, regardless of the type of sediment used in the
suspended particulate phase.
57
-------
PARTICULATE/SOLID PHASE
control/control
burrow length = 39.7 ±9.6
BRH/control
burrow length = 4 1.3 ± 9.8
control/BRH
BRH/BRH
burrow length = 14.9 ±6.8
burrow length - 20.2 ±1.0
Figure 11. Drawing of burrows created by N. arenaceodentata juveniles
exposed to various combinations of reference and BRH
sediment for 10 days
58
-------
Table 16
Burrow length and maximum burrow depth of N. arenaceodentata
juveniles exposed to various sediment conditions
Treatment
REF/REF*
REF/BRH
BRH/REF
BRH/BRH
Burrow Length** Percent in Maximum Burrow Depth**
cm Gpt Sediment mm Gpt
Particulate Phase Assay
39.7 + 9.6 A 49 26.8+11.3 A
14.9 + 6.8 B 24 17.0+ 4.1 B
41.3 + 9.8 A 32 34.2 + 4.0 A
20.2 + 1.0 B 37 17.0 + 9.2 B
* REF • Reference Sediment.
** Means + 1 S.D.
t Gp » Grouping letter. Means having the same Gp letter are not
significantly different at p - 0.05.
59
-------
PART IV: DISCUSSION
70. Nephtys incisa is a nonselective deposit feeder and is
typically found in soft sediments. It does not build permanent
tubes, but rather burrows indiscriminately, ingesting sediment and
associated microorganisms as a food source (Davis 1979). This
species is considered a member of the equilibrium assemblage in
Long Island Sound (Rhoads and Germane 1982), and is usually associated
with sediments that are oxygenated to depths of up to 10 cm. The
physical effects of the errant burrowing behavior of N^ incisa on
local sediments are the provision of vertical particle mixing and
the enhancement of pore water exchange (Rhoads and Germane* 1982).
Much of the burrowing activity occurs at the redox boundary, a zone
typically high in microorganism productivity (Yingst and Rhoads
1980).
71. The data presented here for the reference sediment
treatments indicate that N^ incisa juveniles perform much the same
organism-sediment roles as do the adult stages. Burrowing activity
during the 10-day experimental period was extensive, with burrows
penetrating up to 30 mm in depth. Respiratory pumping also appeared
to be effective in pore water exchange, as the burrow tubes had an
observable 'halo' of light sediment which is an indicator of oxidized
sediment (Aller and Yingst 1978).
72. The physiological measurements determined with Nephtys
incisa juveniles were reproducible and repeatable. (Review Appendix
A for data from each individual experiment.) Within an experiment,
60
-------
each response measured (growth, respiration and ammonia excretion
rate, and burrowing activity) exhibited standard deviations about
the mean value that were less than 28 percent. This is an indication
that the response measurements are reproducible, providing the measure-
ment technique is followed properly. To adequately measure growth,
for example, care must be taken at the beginning of the experiment
to size the worms. Failure to control the variance in initial dry
weight will reduce the chances (statistical) of being able to detect
growth differences among the various treatments at the end of a
10-day experiment.
73. Almost all of the biological endpoints evaluated with
Nephtys incisa followed a dose-response to additions of BRH sediment.
These responses were repeatable from one experiment to another. The
only exception to this was the calculated 0:N ratios which did not
exhibit a response across the treatments used.
74. Net growth efficiencies for N. incisa Juveniles main-
tained in control sediments are within the range of efficiencies
reported for polychaetes under a variety of conditions. For instance,
Carey (1962) estimated population production and respiration values
for N._ incisa from a Long Island Sound study site. Using data
from his Table 9, a mean net growth efficiency of 36 percent is
calculated (population production/population production + respiration),
Net growth efficiency for omnivorous polychaetes (both indiscriminate
sediment ingesters, as well as surface detrital feeders) has been
found to be generally between 14 and 40 percent (Kay and Brafield
1973; Tenore and Gopalan 1974; Neuhoff 1979).
61
-------
75. The effects of exposure to BRH sediment are twofold.
First, N. incisa juveniles greatly reduce their burrowing activity
when exposed to whole BRH sediment. This was graphically demonstrated
in Figures 9 and 10, where worms maintained in BRH sediment for 10
days demonstrated almost no burrowing activity and no depth penetra-
tion. Similar results were found in the lugworm, Arenlcola cristata,
exposed to kepone-contaminated sediments (Rubinstein 1979).
76. In the suspended particulate assays, where a choice
between sediments was offered as settling of the particulate dose
occurred, N. incisa appeared to avoid BRH sediment. Comparison of
the reference/BRH treatment to the BRH/reference treatment illustrates
this point. Almost all of the burrowing activity in the reference/BRH
treatment was in the upper settled layer. This activity occurred
only after the sediment layer was formed. Worms from the BRH/refer-
ence exposure exhibited an opposite activity level. In this treat-
ment, there was extensive reworking of the lower, reference sediment,
while there was very little activity in the upper, sedimented BRH
material. The only burrows present in this upper layer were a few,
fairly vertical ones to the surface (when compared to reference/
reference and reference/BRH). These burrows probably provide the
worms with a supply of oxygenated water to meet their respiratory
requirements.
77. The dramatic decrease in burrowing activity of N^ incisa
juveniles that occurred during exposure to BRH indicates that the
worms are probably curtailing physiological functions. If the
primary energy source of this species is ingested sediment, then
62
-------
curtailing burrowing activity has the effect of starving the indi-
vidual. This is, in essence, the second effect BRH sediment has on
the species. This effect is brought about by the failure of the
worms to consume a sufficient amount of energy for normal physio-
logical and biochemical processes. Nephtys incisa exposed to BRH
sediments generally exhibited lowered maintenance costs, most
probably in response to the reduced energy intake and reduced
burrowing activity. Despite these lowered costs, individuals in
the BRH sediment had to catabolize some tissue in order to provide
the energy requirements of routine metabolism. Catabolism of tissue
to meet energy requirements is a common phenomenon in organisms
that are below their required maintenance ration (Pandian 1975).
78. In those treatments where there was a choice between BRH
and reference sediments, however, there was an increase in burrowing
activity (with a majority of the activity occurring in the reference
sediment; Fig. 10). The increase in activity noted in the BRH/
reference and reference/BRH sediments is apparently sufficient to
provide enough energy for both maintenance costs and growth. Worms
under these sediment conditions had lower, but not significantly
different, net growth efficiencies than did worms maintained in the
reference sediment (Table 11).
79. In contrast to N^ incisa, N^_ arenaceodentata is primarily
a surficial deposit feeder that builds somewhat permanent mucus
tubes near the sediment surface. Unlike N^ incisa, N^ arenaceodentata
will leave their tubes to search the sediment surface for food.
Despite these differences in feeding habits, N^ arenaceodentata
63
-------
juveniles exhibited burrowing patterns similar to those of N^ incisa.
Burrowing activity was reduced in participate phase assays in which
BRH sediment was the bedded sediment. In addition, depth of burrowing
was also shallower in those treatments where BRH sediment existed
as the bedded sediment.
80. The physiological responses of N. arenaceodentata to
exposure to BRH sediment, however, were not as dramatic as they
were with N. incisa. In only one of the three experiments conducted
with ^ arenaceodentata was there a significant difference in dry
weight at the end of the 10-day exposure. In addition, no significant
differences in respiration rates were found in any of the various
experiments. Food consumption rates, however, were found to be
affected, with lower rates found in those worms exposed to BRH
sediment. The general lack of physiological response by N_._
arenaceodentata to BRH sediment may have been due to several factors.
81. One is simply that exposure to BRH sediment does not
cause any change in the growth rate or physiolgical rate of this
species. Unlike N^ incisa, which appears to curtail burrow/feeding
activity in BRH sediment, N. arenaceodentata feeds on detrital
material found on the sediment surface and thus can avoid ingesting
BRH sediment under some conditions. The conclusion that BRH sediment
does not have a physiological effect on N_._ arenaceodentata, however,
is not probable since food consumption rates were affected by exposure
to BRH sediment.
82. It is also possible that the mucus tube formed by this
species may reduce its contact with surrounding sediments. This,
64
-------
in turn, would reduce its exposure to contaminants that are found
in BRH sediment. Although not specifically tested for in this
study, it would be interesting to determine what degree of protection
from sediment contaminants is offered by mucus tubes.
83. Another explanation, however, for the lack of any measur-
able growth response to BRH sediment is that the variance in worm
sizes at the beginning of an experiment was so great that any changes
in growth rate resulting from the treatment exposure could not be
detected after the 10-day exposure period. With N_._ incisa the
coefficient of variation in dry weight at the beginning of an exper-
iment was 24 percent, while with N. arenaceodentata the coefficient
of variation in worm size was quite large in the first two experiments
(42 and 59 percent, respectively). This degree of variation made
it difficult at best to measure growth in as short a time period
(10 days) as used in this study. In the third experiment (particulate
phase, experiment III), on the other hand, where significantly
lowered growth was detected in those worms from the BRH/BRH condition,
the coefficient of variation for dry weight at the start of the
experiment was only 21 percent. While it is not known whether the
relationship between low variation in worm size at the beginning of
an experiment and the ability to detect changes in growth after 10
days is causal or casual, more time will be spent in future experiments
with N. arenaceodentata in selecting worms for experimentation.
84. The net growth efficiencies calculated in this study for
N. arenaceodentata were within the range of values reported in the
literature for detrital feeding polychaetes (Kay and Brafield 1973;
65
-------
Neuhoff 1979). The large variation found about the mean net growth
efficiency in all of these experiments was probably largely due to
poor control in worm size at the beginning of the experiments. It
is interesting to note that the net growth efficiencies of both N.
arenaceodentata and N. incisa in reference sediment conditions were
similar.
85. Insight into differences in mode of exposure that may
exist for these two species can be seen by examining the net growth
efficiencies from the BRH exposures. For Nj,_ incisa, exposure to
BRH sediment greatly reduced burrowing activity which directly
affected the amount of energy (food) ingested. The impact of this
is apparent in the negative net growth efficiency for !*._ incisa
from the BRH treatments.
86. Although burrowing activity is reduced in BRH sediment
with N. arenaceodentata, this fact does not necessarily lead to
reduced energy injestion rates as with N^_ incisa. Since N.
arenaceodentata is a surficial detrital feeder, rates of food
ingestion are probably not related to burrowing activity, at least
not as stongly as it does with sediment injesters such as N. incisa.
Net growth efficiency for KL^ areanaceodentata in BRH/BRH treatment
(experiment II, particulate phase assay) was lower than for those
individuals in the reference/reference condition, but the efficiency
remained positive, suggesting that feeding did continue in BRH
sediment although at some reduced level.
87. Unlike net growth efficiency which is an integrative
measure of past physiological condition (i.e., a measure of the
66
-------
efficiency at which an organism grew during the experimental period),
scope for growth is an index by which the current physiological
condition of an organism is evaluated (i.e., is a measure of the
potential for growth). It does not provide a time course estimate
of the cummulative effects a particular condition has had on an
organism as does net growth efficiency. Rather, SFG evaluates
current physiological condition—the current physiological state
obviously being a product of those conditions to which the organism
was exposed to in the recent past. Combined, these two estimators
of organismal health (net growth efficiency and SFG) offer insights
into what the effects of a particular set of conditions have been.
88. In the present study, the SFG values indicate that BRH
sediment (100-percent BRH, BRH/BRH treatments) reduces the amount
of energy available for growth in N. arenaceodentata juveniles
(Table 14). This is particularly interesting in light of the fact
that the net growth efficiency values for two of the experiments
did not indicate any effect of BRH sediment on the growth of this
species. SFG values of N_._ arenaceodentata followed a dose-reponse
to additions of BRH sediment in all experiments, while net growth
efficiency did not. The reason for this is that SFG values are not
as sensitive to differences in size as is the measure of net growth
efficiency. Changes in size over the experimental period are not
taken into account in SFG determinations whereas these changes in
growth are an important factor in calculating net growth
efficiency.
67
-------
89. The SFG values presented in this study are probably an
overestimate of what may be the actual energy available for growth.
The SFG estimates for worms from the reference sediment exposure
are at least twice as high as the amount of growth that was recorded
during the 10-day experimental period. This was determined by
multiplying the mean SFG value (which is calculated for a 24-hr
period) by the length of the experiment (Table 14) and comparing
this to the average energy equivalent for the amount of tissue
produced during the 10-day exposure (Table 12). This is not an
entirely correct procedure for comparing actual growth with data
which allows for the prediction of growth (in this case, SFG) and
is probably partly responsible for the overestimation of the growth
potential in this particular case. Scope for growth determinations
should instead be made at several times during the experimental
period in which growth will actually be measured to better track
potential for growth. By determining SFG only at the end of the
experimental period, all estimates for the potential for growth are
for individuals larger than existed during most of the experiment.
As the potential for growth probably increases with size (at least
within the juvenile stage), SFG values from worms at the end of the
10-day experimental period should be greater than those from worms
from the beginning of the experiment and would thus tend to over-
estimate previous potential for growth.
90. Another reason to believe that the SFG values presented
here are an overestimate of growth potential is the fact that no
attempt was made to calculate the amount of energy loss associated
68
-------
with mucus production. As noted earlier, N. arenaceodentata builds
tubes of mucus and sediment and these tubes appear to be con-
tinually rebuilt. (Personal observation of the Neanthes cultures
suggest that mucus production is virtually constant.) As mucopoly-
saccharides are high in energy content, the production of these
compounds would be at a considerable cost in terms of energy. For
example, it is estimated that approximately 30 percent of assimilated
energy is partitioned as the cost of mucus production in some inverte-
brate species which produce copious quantities of mucus (Pandian
1975). Future bioenergetics research with this species will take
into account the amount of assimilated energy that is used in mucus
production.
91. The effects of BRH sediment are probably due to at least
one of two factors. Either the worms (especially N. incisa) were
responding to a difference in particle size, or they were reacting
to contaminants within the BRH sediment. While the first possibility
cannot be completely discounted, Carey (1962) found no correlation
between the occurrence of N^ incisa populations in Long Island
Sound and particle-size distribution despite significant differences
in the granulometry among the various sites.
92. The second possibility appears to be the most likely
factor affecting N^ incisa. Black Rock Harbor sediment is a multi-
contaminated sediment that contains a wide variety of anthropogenic
chemicals. Qualitative analysis of BRH sediments indicates the
presence of high concentrations of polychlorinated biphenyls,
polynuclear aromatic hydrocarbons, and heavy metals including Cu,
69
-------
Pb, Cd, and Cr (Rogerson et al. 1984). Despite this large spectrum
of contaminants, BRH sediment is not acutely toxic to N. incisa
juveniles.
93. The exposure conditions presented in this report do not
represent realistic exposure regimes for N_^ incisa in Long Island
Sound. Rather, they were employed to produce a sublethal response.
Current research is aimed at establishing the exposure regimes that
!N^ incisa would be exposed to near the Black Rock Harbor site.
Once these limits are established, long-term laboratory studies
will be conducted with environmental conditions that more closely
approximate field conditions.
70
-------
PART V: CONCLUSIONS
94. The results of this study indicate that the principles of
bioenergetics can be applied to study the effect of sediment disposed
on polychaetes. With Nephtys incisa, the physiological measurements
were found to be reproducible within a particular treatment. The
physiological measurements were also found to be repeatable. In all
instances, measurement values from a particular treatment in one
experiment were not significantly different from the same treatment
in repeated experiments. In most cases, the physiological measures
followed a dose-response to additions of BRH sediment. The only
exception were the 0:N ratios which did not show a response to exposure
to BRH sediment.
95. The findings with Neanthes arenaceodentata were not as
dramatic as they were for N^ incisa. In only one of the three
experiments was there a dose-response to additions of BRH sediment.
71
-------
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Bower, C.E., and Holm-Hansen, T. 1980. "A Salicylate-Hypochlorite
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Capuzzo, J.M., and Lancaster, B.A. 1981. "Physiological Effects
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in Long Island Sound," Ph.D. Thesis, Yale University, New Haven,
Conn.
Conover, W.J., and Iman, R.L. 1981. "Rank Transformations as a
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Corner, E.D.S., and Cowey, C.B. 1968. "Biochemical Studies on
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Davis, W.R. 1979. "The Burrowing, Feeding and Respiratory Activities
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Edwards, R.R.C. 1978. "Effects of Water-soluble Oil Fractions on
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Elliot, J.M., and Davison W. 1975. "Energy Equivalents of Oxygen
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Gilfillan, E.S. 1980. "The Use of Scope-for-growth Measurements
in Monitoring Petroleum Pollution," Biological Effects of Marine
Pollution and the Problems of Monitoring. A.D Mclntyre and J.B.
Pearce, eds., Rapports Et Proces-Verbaux Ces Reunions Conseil Permanent
International Pour L*Exploration Ce La Mer, Vol 179, pp 71-75.
Gilfillan, E.S., and Hanson, S.A. 1975. "Effects of Paralytic
Shellfish Poisoning Toxin on the Behavior and Physiology of Marine
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of Toxic Dinoflagalates, V.R. LoCicero ed., The Massachusetts
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Gilfillan, E.S., Mayo, D., Hanson,S., Donovan,D., and Jiany, L.C.
1976. "Reduction in Carbon Flux in Mya arenaria Caused by a Spill
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Gilfillan, E.S., and Vandermeulen, J.H. 1978. "Alterations in
Growth and Physiology of Soft Shell Clams, Mya arenaria Chronically
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Journal Fisheries' Research Board of Canada, Vol 35(5), pp 630-636.
Goodnight, J.H. 1979. "VARCOMP Procedure" SAS Users Guide. J.T.
Helwig, and K.A. Council, eds-., SAS Institute Inc. Raleigh, N.C.,
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International Council for the Exploration of the Sea. 1978. "On
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Johns, D.M., and Pechenik, J.A. 1980. "Influence of the Water-
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Johns, D.M., and Miller, D.C. 1982. "The Use of Bioenergetics to
Investigate the Mechanisms of Pollutant Toxicity in Crustacean
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W.B. Vernberg, A. Calabrese, F.P. Thurberg, and F.J. Vernberg,
eds.. Academic Press, New York, pp 261-288.
Kay, D.G., and Brafield, A.E. 1973. "The Energy Relations of the
Polychaete Neanthes (=Nereis_) virens (Sars)," Journal of Animal
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Lake, J., Hoffman, G., and Schimmel, S. 1984. "Bioaccumulation of
Contaminants from Black Rock Harbor Dredged Material by Mussels and
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Environmental Protection Agency, Environmental Research Laboratory,
Narragansett, R.I., for the US Army Engineer Waterways Experiment
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Logan, D.T., and Epifanio, C.E. 1978. "A Laboratory Energy Balance
for the Larvae and Juveniles of the American Lobster, Homarus
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Maciolek, J.L. 1962. "Limnological Organic Analysis by Quantitative
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Mclntyrne, A.D., and Pearce, J.B. (eds), 1980. "Biological Effects
of Marine Pollution and the Problems of Monitoring." Rapports Et
Proces-Verbaux Ces Reunions Conseil Permanent International Four
L'Exploration Ce La Her, Vol 179.
McKinney, C.L., Jr. 1982. "Interrelationships Between Energy
Metabolism, Growth Dynamics, and Reproduction During the Life Cycle
of Mysidopsis bahia as Influenced by Sublethal Endrin Exposure,"
Physiological Mechanisms of Marine Pollutant Toxicity. W.B. Vernberg,
A. Calabrese, F.P. Thurberg, and F.J. Vernberg, eds., Academic
Press, New York, pp 447-476.
Neuhoff, H.-G. 1979. "Influence of Temperature and Salinity on
Food Conversion and Growth of Different Nereis Species (Polychaeta,
Annelida)," Marine Ecology Progress Series. Vol 1, pp 255-262.
Pandian, T.J. 1975. "Mechanisms of Heterotrophy," Marine Ecology,
Vol. II. Physiological Mechanisms, Part 1, 0. Kinne, ed. Wiley,
Chichester, pp 61-250.
Pesch, C.E., and Hoffman, G.L. 1983. "Interlaboratory Comparison
of a 28-day Toxicity Test With the Polychaete Neanthes arenaceodentata,
Aquatic Toxicology and Hazard Assessment; Sixth Symposium, ASTM
STP 802. W.E. Bishop, R.D. Cardwell and B.B. Heidolph, eds., American
Society for Testing and Materials, Philadelphia, pp 482-493.
Rhoads, D.C., and Germane, J.D. 1982. "Characterization of Organism-
sediment Relations Using Sediment Profile Imaging: An Efficient
Method of Remote Ecological Monitoring of the Seafloor (Remots (TM)
System)," Marine Ecology Progress Series, Vol 8, pp 115-128.
Rogerson, P., Schimmel, S., and Hoffman, G. 1984. "Chemical and
Biological Characterization of Black Rock Harbor Dredged Material,"
Technical Report D-84-, prepared by U.S. Environmental Protection
Agency, Narragansett, R.I., for the U.S. Army Engineer Waterways
Experiment Station, CE, Vicksburg, Miss.
Rubinstein, N. 1979. "A Benthic Bioassay Using Time-lapsed Photog-
raphy to Measure the Effect of Toxicants of the Feeding Behavior
of Lugworms (Polychaeta: Arenicolida)," Marine Pollution; Functional
Responses. W.B. Vernberg, F.P. Thurberg, A. Calabrese, and F.J.
Vernberg, eds.. Academic Press, New York, pp 341-351.
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Sanders, H.L. 1956. "Oceanography of Long Island Sound. X. The
Biology of Bottom Marine Communities," Bulletin Binghamton Oceanograghy
Collection, Vol 15, pp 345-414.
Sanders, H.L. 1958. "Benthic Studies in Buzzards Bay. I. Animal-
sediment Relationships," Limnology and Oceanography, Vol 3(3),
pp 245-253.
Schauer, P.S., and Pesch, C.E. In Prep. "Influence of Diet on
Growth, Survival and Tube Production of Laboratory Cultured Polychaete
Neanthes arenaceodentata."
Snedecor, G.W., and Cochran, W.G. 1980. "Statistical Methods,"
Iowa State University Press, Ames.
Tenore, K.R., and Gopalan, U.K. 1974. "Feeding Efficiencies of
the Polychaete Nereis virens Cultured on Hard-clam Tissue and Oyster
Detritus," Journal Fisheries Research Board of Canada, Vol 31,
pp 1675-1678.
Warren, C.E., and Davis, G.E. 1967. "Laboratory. Studies on the
Feeding, Bioenergetics and Growth of Fish," The Biological Basis of
Freshwater Fish Production, S.D. Gerking, ed., Blackwell, Oxford,
pp 175-214.
Yingst, J.Y., and Rhoads, D.C. 1980. "The Role of Bioturbation in
the Enhancement of Mictobial Turnover Rates in Marine Sediments,"
Marine Benthic Dynamics, K.R. Tenore and B.C. Coull, eds., University
of South Carolina Press, Columbia, pp 407-421.
75
-------
APPENDIX A: SOLID PHASE AND PARTICULATE PHASE DATA SHEETS
Al
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY PLAN: 3
EXPERIMENT DESCRIPTION: SOLID
INVESTIOATOR: JOHNS/GU T JAHR
DATE OF TEST: 821014
SPECIES: NEPHTYS INC ISA
** EXPERIMENTAL CONDITIONS **
TEMPERATURE: 21. OO DEGREES CENTIGRADE
SALINITY: 28. OO PARTS PER THOUSAND
EXPOSURE DURATION: 1O DAYS
PHOTOPERIOD: 13 HOURS
FLOW RATE: SO MLS/MIN
NUMBER OF ANIMALS/REPLICATE:
ANIMAL'S LIFE STAGE: JUVENILE
SIZE: 1.39 +/- 0.21 MG DRY WT
CONTROLS: 1OO PERCENT REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: SOUTH REFERENCE, LONG ISLAND SOUND
COLLECTION TEMPERATURE: 19. 3 C
ACCLIMATION: 20 C.
SEDIMENT SCUHCE: BARREL OR COLLECTION/JAR NUMBER
SOLID REFERENCE: 11/SO SOLID BRH: LL/23
SUSPENDED REFERENCE: SUSPENDED BRH:
RANGE: 2O.30 - 21. SO
RANGE: 28. OO - 29. OO
VOLUME ADDITIONS/DAY:
13 NUMBER OF REPLICATES/TREATMENT:
COLLECTION SALINITY: 29
SAMPLE: EXPOSURE CONCENIRATIONS!
GROWTH
JOULES
NUMBER
!RESPIRATORY!EXCRETORY !NET GROWTH
! ENERGY ! ENERGY !EFFICIENCY
NOMINAL !MEASURED! ! !EXPENDITURE! LOSS !IN PERCENT
! ! ! i JOULES ! JOULES ! !
! !MEAN +/-STD !MEAN+/-STD 1MEAN+/-STD1MEAN+/-STD
400020 100 PERCENT REF!
400021 30:30 REF:BRH !
!
400022 100 PERCENT BRH!
! . 87 ! . O7 i 6. 1 ! 1. 8 i 0. O4!O. 01 ! 17 ! 10
i ;:::;: i
! -. 34 ! . 11 1 3. 3 ! 2. 1 ! 0. O3!0. Ol! -3 ! 14
! !!!!!! i
!-3. 33 ! . 34 ! 6. 3 ! 2. 8 ! 0. 02!0. 01! -6O ! 20
A3
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY FLAN: 3
EXPERIMENT DESCRIPTION: SOLID
INVESTIGATOR: JOHNS/CUTJAHR
DATE OF TEST: 821203
SPECIES: NEPHTYS INC ISA
** EXPERIMENTAL CONDITIONS »»
TEMPERATURE: 2O. 30 DEGREES CENTIGRADE
SALINITY: 30. 00 PARTS PER THOUSAND
EXPOSURE DURATION: 10 DAYS
PHOTOPERIOD: 13 HOURS
FLOW RATE: 50 KLS/MIN
NUMBER OF ANIMALS/REPLICATE:
ANIMAL'S LIFE STAGE: JUVENILE
SIZE: 1.4O +/- O. 33 MG DRY WI
CONTROLS: 1OO PERCENT REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: SOUTH REFERENCE, LONG ISLAND SOUND
COLLECTION TEMPERATURE: 6. 7 C
ACCLIMATION: 2O C.
SEDIMENT SOURCE: BARREL OR COLLECTION/JAR NUMBER
SOLID REFERENCE: 11/50 SOLID BRH: LL/25
SUSPENDED REFERENCE: SUSPENDED BRH:
RANGE: 2O.OO - 21. 5O
RANGE: 28. 5O - 31. OO
VOLUME ADDITIONS/DAY:
15 NUMBER OF REPLICATES/TREATMENT:
COLLECTION SALINITY: 29
SAMPLE! EXPOSURE CONCENTRATIONS!
NUMBER!
!
!
NOMINAL
GROWTH !RESPIRATORY{EXCRETORY ! NET GROWTH
! JOULES ! ENERGY ! ENERGY !EFFICIENCY
MEASURED! ! !EXPENDITURE! LOSS !IN PERCENT
! i i JOULES i JOULES i !
!MEAN +/-STD SMEAN+/-STD !MEAN+/-STD!MEAN-t-/-STD
400023!100 PERCENT REF
I
I
40OO23!5O:5O REF:BRH
I
f
40OO27J100 PERCENT BRH
5 . 87! . 47 i 13. 5! 3. 7 ! 0. O4!0. 01! 30 ! 8
!!!!!! !
4. 13 ! 1.2 ! 11.31 2.9 ! 0. O3:O.Oi: 26 ! 7
I J I « « t I
-O. 04 ! .0 ! 5.3! 1.2 ! 0. O2!0. 01! -1 ! 4
!!!!!! i
A4
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY PLAN: 3
INVESTIGATOR: JOHNS /GUTJAHR
EXPERIMENT DESCRIPTION: SOLID
DATE OF TEST:
821203
SPECIES: NEPHTYS INC ISA
»•* EXPERIMENTAL CONDITIONS **
RANGE: SO.OO - 21. 30
RANGE: 28.90 - 31. OO
TEMPERATURE: 20.50 DEGREES CENTIGRADE
SALINITY: 30. OO PARTS PER THOUSAND
EXPOSURE DURATION: 1O DAYS
PHOTOPERIOD: 13 HOURS
FLOU RATE: SO I1LS/MIN
NUMBER OF ANIMALS/REPLICATE:
ANIMAL'S LIFE STAGE: JUVENILE
SIZE: 1.43 +/- 0.73 MG DRY WT
CONTROLS: 1OO PERCENT REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: SOUTH REFERENCE
COLLECTION TEMPERATURE. 6. 7 C
ACCLIMATION: 20 C.
SEDIMENT SOURCE: BARREL OR COLLECTION/JAR NUMBER
SOLID REFERENCE: 11/50 SOLID BRH: LL/23
SUSPENDED REFERENCE: SUSPENDED BRH:
VOLUME ADDITIONS/DAY:
13 NUMBER QF REPLICATES/TREATMENT:
LONG ISLAND SOUND
COLLECTION SALINITY: 29
SAMPLE! EXPOSURE CONCENTRATIONS!
! ! !
NUMBER! NOMINAL ! MEASURED i
! ! !
! ! !
GROV
JOUl
MEAN +i
400036! 100 PERCENT REF! ! 4.04
! ! i
400O37!5O:30 REF: BRH ! ! O. 65
! ! !
400O38J1OO PERCENT BRH! !-l. 85
! ! !
•ITH ! RESPIRATORY! EXCRETORY
.ES ! ENERGY ! ENERGY
{EXPENDITURE! LOSS
! JOULES ! JOULES
'-STD SMEAN+/-STD !MEAN+/-STD
. 73
. 16
. 17
11. 8! 2. 5 ! 0. 04!0. 01
i i !
6. 2! 1.3 i O. O310. Ol
III
3. 3! 2. 3 ! 0. 0210. 01
! ! !
NET GROWTH
EFFICIENCY
IN PERCENT
!
MEAN+/-STD
23 i 5
!
1O ! 2
1
-24 ! 14
!
A5
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY PLAN: 3 INVESTIGATOR: JOHNS/CUTJAHR
EXPERIMENT DESCRIPTION: SUSPENDED DATE OF TEST: 830902
SPECIES: NEPHTYS INC ISA
** EXPERIMENTAL CONDITIONS *»
TEMPERATURE: 21.20 DEGREES CENTIGRADE
SALINITY: 30. SO PARTS PER THOUSAND
EXPOSURE DURATION: 10 DAYS
PHOTOPERIOD: 13 HOURS
FLOW RATE: 35 MLS/MIN
NUMBER OF ANIMALS/REPLICATE:
ANIMAL'S LIFE STAGE: JUVENILE
SIZE: 3. 13 +/- 0.60 MO DRY WT
CONTROLS: 2OO MG/L REF/REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: SOUTH REFERENCE/ LONG ISLAND SOUND
COLLECTION TEMPERATURE: 2O. 3 C
ACCLIMATION: 20 C.
SEDIMENT SOURCE: BARREL OR COLLECTION/ JAR NUMBER
SOLID REFERENCE: 1 1 1/19 SOLID ERH: EE/3, 8
SUSPENDED REFERENCE: 111/19,21,22 SUSPENDED BRH: EE/7, 11,12
RANGE: 20. 30 - 22. SO
RANGE: 30.OO - 31. OO
VOLUME ADDITIONS/DAY: 67
IS NUMBER CF REPLICATES/TREATMENT:
COLLECTION SALINITY: 25 8
SAMPLE! EXPOSURE CONCEf
NUMBER! NOMINAL
40O1301200MG/L REF/REF
1
400131 I200MG/L BRH/REF
I
1
40Q132S200MG/L REF/BRH
400133 S200MS/L BRH/BRH
v'TRATIONS
MEASURED
211+87
171+53
211+87
171+33
GRO'v
JOUt
MEAN +/
6. 09
4. 35
3. OO
-2. 30
^TH
-ES
'-STD
1.22
. 96
1. 1O
. 78
RESPIRATORY
ENERGY
EXPENDITURE
JOULES
MEAN+/-STD
14. 1
13. 7
10. 2
9. 0
4. 7
3. 2
4. 7
3. 3
EXCRETORY
ENERGY
LOSS
JOULES
MEAN+/-STD
O. O8!O. 01
1
O. O8!0. O2
O. O3!0. 01
1
0. 0410. 01
NET GROWTH
EFFICIENCY
IN PERCENT
MEAN+/-STD
35 ! 5
1
1
24 ! 3
36 ! 22
-78 ! 48
A6
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY PLAN: 3
INVESTIGATOR: JOHNS/OUFJAHR
EXPERIMENT DESCRIPTIO:4: SUSPENDED
DATE OF TEST:
830920
SPECIES: NEPHTYS INC ISA
** EXPERIMENTAL CONDITIONS •**
RANGE: 19. SO - 22. GO
RANGE: 30.00 - 31. 8O
TEMPERATURE: 2O. 60 DEGRESS CENTIGRADE
SALINITY: 3O.70 PARTS PER THOUSAND
EXPOSURE DURATION: 10 DAYS
PHOTOPERIOD: 13 HOURS
FLOW RATE: 32 fS-S/HIN VOLUilE ADDITIONS/DAY: 61
NUM3ER OF ANIMALS/REPLICATE: 15 NUMBER OF REPLICATES/TREATMENT: 1
ANIMAL'S LIFE STAGE: JUVENILE
SIZE: 3.43 +/- 0.92 KG DRY Wf
CONTROLS: 2CO KG/L REF/REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: SOUTH REFERENCE, LONG ISLAND SOUND
COLLECTION TEMPERATURE: 21.6 C COLLECTION SALINITY: 29.2
ACCLIMATION: 2O C.
SEDIMENT SOUHCE: BARREL OR COLLECTION/JAR NUMBER
SOLID REFERENCE: 111/23 SOLID BRH: EE/17
SUSPENDED REFERENCE: HI/6, 7, 36 SUSPENDED BRH: EE/8. 10, 14
SAMPLE! EXPOSURE CONCENTRATIONS! GROWTH
! i JOULES
NUMBER ! NOM I NAL ! MEASURED :
1
1
4OG134I20OKG/L REK/REF
I
1
400133 J20CKS/L BRH/REF
!
4O013612OCKG/L REF/BRH
j
40O13712OOMO/L BRH/BRH
!
(MEAN +/
193+73 !13. 38
!
226+48 i 8 48
t
I
193+73 ! 5. 11
j
226+48 i 1. 41
r-STD
. 94
1. 36
. 92
RESPIRATORY
ENERGY
EXPENDITURE
JOULES
MEAN+/-STD
17. 8! 4.4
t
18. 3! 3. 8
t
1
12. 2! 2. 8
1
.27 ! 10. 6! 2. 3
EXCRETORY
ENERGY
LOSS
JOULES
MEAN+/-STD
O. 0710. 01
1
1
0. 07 !0. 03
1
1
O. 07 !0. 02
j
0. 0510. 01
NET GROWTH
EFFICIENCY
IN PERCENT
MEAN+/-
44
33
29
•STD
7
7
4
12 ! 2
A7
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY PLAN: 3
EXPERIMCNT DESCRIPTION: SOLID
INVESTIGATOR: JOHNS/GUTJAHR
DATE OF TEST: 830516
SPECIES: NEANTHES ARENACEODENTATA
** EXPERIMENTAL CONDITIONS **
TEMPERATURE: 21. OO DEGREES CENTIGRADE
SALINITY: 28. OO PARTS PER THOUSAND
EXPOSURE DURATION: 1O DAYS
PHOTOPERIOO: 13 HOURS
FLOW RATE: 100 HLS/MIN
NUMBER OF ANIMALS/REPLICATE:
ANIMAL'S LIFE STAGE: JUVENILE
SIZE: 4. 16 + /- 1.73 MS DRY WT
CONTROLS: 100 PERCENT REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: CULTURE
COLLECTION TEMPERATURE:
ACCLIMATION: 20 C.
SEDIMENT SOURCE:
SOLID REFERENCE: I I/SO
SUSPENDED REFERENCE:
RANGE: 20. 30 - 21. 30
RANGE: 28. 00 - 29. 00
VOLUME ADDITIONS/DAY 67
13 NUMBER OF REPLICATES/TREATMENT:
AGE: 41 DAYS
COLLECTION SALINITY:
BARREL OR COLLECTION/JAR NUMBER
SOLID BRH: LL/23
SUSPENDED BRH:
SAMPLE! EXPOSURE CONCENTRATIONS
! i
NUMBER! NOMINAL {MEASURED
!
'•
4OOO23S10O PERCENT REF
!
400029! 50: 50 REF: BRH
j
400030 110O PERCENT BRH
! !
GROWTH
JOULES
I
1
t
MEAN -»-/-STD
22 ! 20
!
14 ! 19
!
13 ! 14
1
RESPIRATORY
ENERGY
EXPENDITURE
JOULES
MEAN+/-STD
38 ! 9
!
30 ! 11
1
23 i 3
EXCRETORY !NET GROWTH
ENERGY {EFFICIENCY
LOSS ! IN PERCENT
JOULES i !
MEAN+/-STD ! MEAN+/-STD
0. 06 { 0. O2 ! 29 ! 34
t t <
1 1 I
0.03! 0.02! 18 ! 42
! ! !
0. 04 !0. 01! 27 ! 32
! ! ! ! i
A8
-------
LABORATORY WORM DATA SHEET
COE/ERLN FVP
STUDY PLAN: 3
INVESTIGATOR: JOHNS/GUTJAHR
EXPERIMENT DESCRIPTION: SUSPENDED
DATE CF TEST:
830*722
SPECIES: NEANTHES ARENACEOCENTATA
*+ EXPERIMENTAL CONDITIONS **
RANGE: 19, 80 - 22. OO
RANGE: 3O. OO - 31. 80
TEMPERATURE: 2O. 6O DECREES CENTIGRADE
SALINITY: 30. 70 PARTS PER THOUSAND
EXPOSURE DURATION: 10 DAVS
PHOTOPERIOD: 13 HOURS
FLOW RATE: 33 MLS/MIM VOLUME ADDITIQNS/'DAY 63
NUMBER OF ANIMALS/REPLICATE: 13 NUMBER CF REPLICATES/TREATMENT:
ANIMAL'S LIFE STAGE: JUVtNILE AGE: 42 DAYS
SIZE: 3. 52 +/- 2.09 MG DRY WT
CONTROLS: 20O MG/L REF/REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: CULTURE
COLLECTION TEMPEKATURE: COLLECTION SALINITY:
ACCLIMATION: 20 C.
SEDIMENT SOURCE: BARREL OR COLLECTION/JAR NUMBER
SOLID BRH: EE/17,18
SOLID REFERENCE: 111/23,26
SUSPENDED REFERENCE: 111tb, 7,36
SUSPENDED BRH: EE/8. 1O, 14
SAMPLE !
!
NUMBER !
1
4O0133!
t
4O0139!
1
4O0140!
i
400141!
EXPOSURE CONCENTRATIONS!
NOMI
20CMG/L
20CMO/L
20CM3/L
20CKO/L
MAL
RE»=YREF
BRH/REF
REF/BRH
1
GROWTH J RESPIRATORY
JOULES ! ENERGY
MEASURED i
199
222
199
3RH/BRHS222
1
1
1
+ 73!
1
+ 44!
1
1
+ 73!
1
+ 44!
MEAN
39
49
43
37
I ! EXPENDITURE
! ! JOULES
+/-STD !MEAN-t-/-STD
: 38
1
5 40
•*
i 27
37
34
61
f 1
t 1
13
13
3
! 32 5 62 ! 7
EXCRETORY 1NET
ENERGY !
LOSS i
JOULES !
KEAN+/-STD!
0. 08 !O. O4!
i ;
O. 14!0. O3!
! i
O. 1110.03!
t I
0. 13J0.07!
GROWTH
EFFICIENCY
IN PERCENT
5
MEAN-f/-STD
3O
41
39
31
! 27
*
; 20
j
i 16
1
! 23
A9
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LABORATORY WORM DATA SHEET
CCE/ERLN FVP
STUDY PLAN: 3 INVESTIGATOR: JOHNS/CUTJAHR
EXPERIMENT DESCRIPTION: SUSPENDED DATE OF TEST:
830816
SPECIES: NEANTHES ARENACEODENTATA
-*•» EXPERIMENTAL CONDITIONS **
RANGE: 20. SO - 21. 50
RANGE: 30.00 - 31. 00
TtKPERATUSE: 21. OO DEGREES CENTIGRADE
SALINITY: 30. 00 PARTS PER THOUSAND
EXPOSURE DURATION: 10 DAYS
PHOTOPERIOD: 13 HOURS
FLOW RATE: 33 HLS/MIN VOLUME ADDITIONS/DAY 67
NUMBER OF ANIMALS/REPLICATE: 13 NUMBER OF REPLICATES/TREATMENT:
ANIMAL'S LIFE STAGE: JUVtNILE AGE: 37 DAYS
SIZE: 4. 43 +/- 0. 092 MG DRY WT
CONTROLS: 200 MG/L REF/REF
FOOD: PRAWN FLAKE SUSPENSION
ANIMAL SOURCE: CULTURE
COLLECTION TEMPERATURE: COLLECTION SALINITY:
ACCLIMATION: 20 C.
SEDIMENT SOURCE: BARREL OR COLLECTION/JAR NUMBER
SOLID BRH: EE/1.2
SUSPENDED BRH: EE/l-3
SOLID REFERENCE: 111/13,14
SUSPENDED REFERENCE: 111/13-17
SAMPLE: EXPOSURE CONCENTRATIONS
NUMBER: NOMINAL [MEASURED
40O126I20CMG/L RE*-/REF!217 + 86
1 1
400127S20CMG/L RE*-/BRH!190 + 61
400123!20CKS/L BRH/REFI217 + 86
t i
40O129S2OCMC/L BRH/BRH!190 + 61
GRO;
JOUl
MEAN -»•
36
38
3
4TH
.ES
'-STD
33
39
31
14
RESPIRATORY
ENERGY
EXPENDITURE
JOULES
MEAN* /-STD
52
36
33
47
6
10
6
9
EXCRETORY !NET GROWTH
ENERGY ! EFFICIENCY
LOSS ! IN PERCENT
JOULES ! !
MEAN+/-STD ! MEAN-t-/-STD
0.08! 0.02! 33 ! 19
i ! !
O. 08 JO. 03! 34 ! 24
ft i
O. 14!0. 03! 33 ! 20
1 f 1
0. 11 SO. 03! 3 ! 23
A10
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