20050785 Ver _Information Letter_20090416

a
STATE OF NORTH CAROLINA

DEPARTMENT OF TRANSPORTATION ,1.
MICHAEL F. EASLEY LYNDO TIPPE I`i '
GovERNOR SECRETARY
April 13, 2009
U.S. Army Corps of Engineers NCDENR Division of Coastal Management
Regulatory Field Office 400 Commerce Ave
Post Office Box 1000 Morehead City, NC 28557
Washington, North Carolina 27889-1000
N. C. Dept of Env. and Natural Resources
Division of Water Quality
Washington District Office O
943 Washington Square
Washington, North Carolina 27889
ATTN: Mr. William Wescott, USACE NCDOT Coordinator
Mr. Steven Lane, DCM NCDOT Coordinator
Mr. Garcy Ward, DWQ NCDOT Coordinator
Dear Sirs:
Subject: Submittal of Revised In-Water Work Study Plan for Review and
Approval per Permit Conditions of 404 Individual Permit, 401 Water
Quality Certification Permit, and Letter of Refinement for CAMA
Major Development Permit for the proposed Washington Bypass. US 17
from South of SR 1149 (Price Road.) to US 17 North of SR 1509 (Springs
Road.). TIP Number: R-251013, Federal Aid Project NHF-17-(54).
Reference: USACE Action ID 199301143
DWQ Project No. 050785, WQC No. 3527
The North Carolina Department of Transportation (NCDOT) is submitting a revised study
plan to monitor the effects of in-water pile driving work on anadromous fishes per the
revised permit conditions. The study plan submitted on March 19, 2009 via email has
been revised based on verbal comments from a meeting held on March 23, 2009 and
written comments forwarded to the Department and the Design-Build Team via email
from March 30, 2009 through April 6, 2009.
NCDOT will also strictly adhere to requirements of the permit modification, in relation to
the proposed study, concerning the mortality rates of certain fish species. If the mortality
of any of the following fish species with respective numbers is observed, then all in-water
work shall cease pending further evaluation by the applicable permitting agencies:
Sturgeon- I fish for the entire in-water work period, and/or 25 fish per bent installation in
any combination of the following species: River herring, Hickory Shad, American Shad,
or Striped Bass.
MAILING ADDRESS: TELEPHONE: [phone] LOCATION:
NC DEPARTMENT OF TRANSPORTATION FAX: [phone] TRANSPORTATION BUILDING
PROTECT DEVELOPMENT AND ENVIRONMENTAL ANALYSIS 1 SOUTH WILMINGTON STREET
1548 MAIL SERVICE CENTER WEBSITE: WWWNCDOTORG RALEIGH NC
RALEIGH NC 27899-1548
April 13, 2009
Page 2
The NCDOT respectfully requests that the referenced study be reviewed as soon as
possible. We are providing copies of this study to all applicable resource agencies
directly.
If you have any questions or need additional information, please call me at (252) 830-
3495.
Sincerely,
Mana A. Rogerson,PE
Assistant Resident Engineer
cc: W/attachment
Mr. Travis Wilson, NCWRC
Mr. Pete Benjamin, USFWS
Mr. Gary Jordan, USFWS
Mr. Louis B. Daniel, III, NCDMF
Mr. Ron Sechler, NCDMF
Mr. Steve Sollod, NCDCM
Mr. Brian Wrenn, NCDWQ
Mr. Chris Underwood, PDEA
Mr. C. E. Lassiter, Jr., P.E., NCDOT
Mr. Rodger Rochelle, PE, NCDOT
Mr. Ron Hancock, PE, NCDOT
Mr. Ed Eatmon, PE, NCDOT
Mr. Mike Robinson, PE, NCDOT
Mr. Jay Johnson, NCDOT
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Assessing the Effects of Bridge Construction Pile Driving Events on
Anadromous Fish Species: NCDOT TIP Project R-2510B, Washington Bypass
Revised Research Proposal to
Mark Mallett
Project Manager
Washington Bypass Project
Flatiron/United JV
2895 US Highway 17 South
Chocowinity, North Carolina 27817
Contract Dates: April 10, 2009- April 9, 2010
Total Project Costs: $481, 675
Prepared by:
Roger A. Rulifson, Ph.D.
Senior Scientist, Institute for Coastal Science and Policy
Professor, Department of Biology
Joseph J. Luczkovich, Ph.D.
Associate Scientist, Institute for Coastal Science and Policy
Associate Professor, Department of Biology
Mark W. Sprague, Ph.D.
Associate Professor, Department of Physics
Anthony S. Overton, Ph.D.
Assistant Professor, Department of Biology
Patrick J. Harris, Ph.D.
Instructor, Department of Biology
East Carolina University
Flanagan 250
Greenville, NC 27858
Revised April 8, 2009
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BACKGROUND
The contractor for NCDOT TIP Project R-22510B, Washington Bypass (Flatiron/United), has
requested a variance to the construction moratorium from February 15 to June 15. The
moratorium falls under the USACE 404 Permit, the DWQ 401 Water Quality Certification, and
the DCM Major Development Permit. This moratorium prohibits in-water work in the Tar-
Pamlico River just upstream of the current U.S. Highway 17 Bridge at Washington during the
springtime anadromous fish spawning run (Figure 1). If the variance is granted, the contractor
agrees to restrict pile driving activity to four hours one day per week, with those four hours
spread over the day.
Figure 1. Completed pile driving event for the first bent crossing over the Tar-Pamlico River,
February 2009.
Each pile driving event takes about 20-30 minutes. There are 10 or 12 concrete pilings spaced
every eight feet for a section of the bridge, referred to here as a "bent" (Figure 1). If the
moratorium is enforced until June 15, Flatiron/United estimates that bridge construction could
take an additional year.
Seven agencies are involved in resolving this issue - The U.S. Army Corps of Engineers, the
U.S. Fish and, Wildlife Service, NOAA Fisheries, the NC Division of Marine Fisheries, the NC
Wildlife Resources Commission, the NC Division of Water Quality, and the NC Division of
Coastal Management. All agencies agree that, if the moratorium is lifted, then field research
must be conducted during construction to determine potential effects of pile driving and in-water
work on the anadromous fish species spawning run. The species in question are the striped bass
Moron saxatilis, American shad Alosa sapidissima, hickory shad Alosa mediocris, alewife
Alosa pseudoharengus, blueback herring Alosa aestivalis, and Atlantic sturgeon Acipenser
oxyrhynchus oxyrhynchus. The Atlantic sturgeon is considered a species of special concern.
Alewife and blueback herring, collectively referred to as "river herring", are species of special
concern in North Carolina. There may be limits for mortality placed in this variance by the
regulatory agencies to determine cessation.
GOAL AND OBJECTIVES
Goal of the study proposed herein is to provide information about anadromous fish passage
during the moratorium variance period, and to determine potential effects of pile driving on
various life history stages of these key species. Specific objectives are:
1. To determine the pattern of sound propagation in the Tar River during pile driving
events;
2. To determine the effects of pile driving on fish movement, specifically upstream
migration of anadromous species, during pile driving events;
3. To estimate rate of injury and death of pile driving events on adult anadromous fish
species, primarily hickory shad, American shad and striped bass;
4. To determine the extent of larval fish passage downstream during the pile driving events;
5. To estimate rate of injury and death of larval fish species during pile driving events.
These five objectives will be broken down into six field research activities, all of which overlap
and most will be performed concurrently.
Job 1 - Pile Driving Sound Propagation Measurements (Sprague)
Job 2 - Adult Fish Upstream Passage (Luczkovich)
Job 3 - Adult Fish Identification and Relative Abundance during Upstream Passage and
Mortality Survey (Rulifson)
Job 4 - Fish Injury and Mortality (Overton, Sprague, Harris, Krahforst)
Job 5 - Larval Fish Downstream Migration (Rulifson)
Job 6 - Larval Fish Injury and Mortality (Overton and Sprague)
In order to be ready to perform the measurements for Jobs 1-6, we must have a minimum of
three weeks from the time the contract is finalized in order to purchase supplies and assemble
our equipment and to make background sound measurements and fish passage baseline
measurements at the study site. It is expected that Flatiron/United will coordinate the pile
driving schedule with the researchers to allow us adequate time to prepare to take measurements
before pile driving begins on each bent and to move our equipment into position before driving
each piling. We cannot work during the night, especially Job 3, which requires visual sighting of
floating fishes as required by the agencies listed above. This aspect would be infeasible in the
dark Assuming April 10 for a contract, that puts us at May I for initiating pile driving
Job 1 - Pile Driving Sound Propagation Measurements (Sprague)
The major concern with a variance on the moratorium is the potential damage or death to
resident and anadromous fish species, and potential behavioral changes that might result from
exposure to the noise of pile driving events. The Tar-Pamlico River at the site of bridge
construction is typical of a lower portion of coastal watershed: wide and slower moving waters
brackish or fresh depending on upstream flow rates, with a narrow channel and wider shallow
shorelines. Shallow areas typically have submerged and emergent vegetation combined with
submerged and emergent tree stumps.
Predicting sound propagation in shallow water can be difficult due to many factors that affect the
sound waves. Sound reflects from the water surface and the bottom and scatters from any
irregularities in the bottom profile. Sound propagating in shallow water has a cutoff frequency,
below which the sound waves decay exponentially. For hard bottoms, the cutoff frequency is the
frequency with wavelength equal to four times the water depth (i.e., the depth is'% wavelength),
but the cutoff frequency is greater for soft bottoms such as sand. The cutoff frequency for 5 in
(16 ft) deep water over a sandy silt bottom is approximately 220 Hz, and for 1 m (3 ft) deep
water over the same bottom is approximately 1,100 Hz. Sound also propagates in the substrate
reflecting off of underground layers with different acoustic properties and passing back into the
water. At frequencies below the cutoff frequency most of the sound propagating significant
distances from the source propagates in the substrate. What this means for the Tar River site in
question (which is very shallow <l m, except in the channel which is 3 m) is that sound
frequencies above 1100 Hz will propagate in the shallow water, and frequencies above 220 Hz
will propagate in the channel. Sound frequencies below the cutoff frequency will likely
propagate in the riverbed and reflect back into the water. The sound will have peak pressure
around 200 dB re 1 µPa at the 10 m distance from the pile. Sound exposure levels near 170-180
dB re 1 µPa 2-s1 and above are expected, with most of that energy in the frequencies below 1000
Hz (Hastings and Popper 2005). The frequency spectrum will have a peak sound exposure
spectral density of approximately 170 dB re 1 µPa 2-s 1 /Hz near 200 Hz and will drop off to 140
dB re 1 µPa 2-s l /Hz above 1000 Hz. Particle velocities will also increase at the onset of each
strike, which will vibrate the otoliths (ear bones) of the exposed fish, and other internal cavities,
potentially causing internal hemorrhaging and death. Little work has been done to study these
effects on fishes (see Hastings and Popper 2005 for a review for CalTrans on Pacific species),
and no work on the species in question here. A further complication is that particle velocities in
a sound wave are maximum at the water surface. Fish in shallow water are likely to be near the
surface and exposed to higher particle velocities than fish at greater depths in deep water.
Before construction begins, sound levels will be recorded at the construction site for a 24-hour
period to establish background sound levels. Pile driving will happen in 20 min periods of
hammering, then a short break occurs to set up for another piling (-1.5 h), then the second pile is
driven; there are 12 piles per bent (bridge section). Preliminary sound propagation
measurements will be conducted during the pile driving at Bent 94 using three hydrophones and
three small boats. A reference hydrophone will be placed approximately 10 m (30 ft) from Bent
94 and will be connected to a digital recorder on a small boat anchored in the river. The
reference hydrophone will make a continuous recording during all of the pile driving events,
which will serve as a reference for recordings made with the other hydrophones. Hydrophones
will be deployed from two additional boats, which will move to different sites during each of the
12 pile driving events during pile driving for Bent 94. The sound recordings at these 25 sites (see
Table 1) will allow us to make a sound field map for the pile driving at the bent. Sound
recording sites will be closer together near the bent site and farther apart at greater distances
where the sound levels are likely to be much lower and will decay less rapidly. A similar sound
recording pattern will be used for pile driving at each bent during bridge construction, but the
deployment pattern will be optimized to obtain maximum sound field information for the pile-
driving sound produced at that location.
In addition to using hydrophones to measure the sound pressure, a sound particle velocity probe
will be constructed using an encapsulated accelerometer. This probe will be deployed at the 10-
m reference site to measure the particle velocity at that location. This instrument must be
custom-built and will not be available during the initial pile driving. Particle velocities from the
initial pile driving locations will be estimated using sound pressure measurements in propagation
models. The accuracy of this use of propagation modeling to estimate particle velocities will be
verified using later particle velocity measurements.
Table 1. Hydrophone sites for measuring sound levels during pile-driving at Bent 94. Angles
are given with respect to a transect line across the river perpendicular to the bank. Positive
distances indicate positions toward the opposite (south) river bank, and negative distances refer
to positions toward the near (north) river bank. The,hydrophone at 10 m along the 0° line will
record sound during all pile driving events and will serve as a reference hydrophone for the other
sound measurements.
90" downstream 20, 30, 50
45° upstream 20, 30, 50
0° -30, -20, -10, 10 (reference), 20, 30, 40, 50, 75, 100, 200, 300, 400
45° downstream 20, 30, 50
90° downstream 20, 30, 50
Job 2 - Adult Fish Upstream Passage (Luczkovich with consulting BioSonics, Inc.
scientists, Eric Munday, BioSonics team leader).
Adult fish frequenting the construction area of the Tar-Pamlico River will be counted and their
direction of travel will be determine using hydroacoustic methods (scientific echosounders). The
anadromous fish migrating through the area during the period of the in-water construction
moratorium include American shad (Alosa sapidissima), hickory shad (Alosa mediocris), river
herrings (Alosa pseudoharengus and A. aestivalis), Atlantic sturgeon (Acipenser oxyrhynchus),
and striped bass (Morone sazatilis).
Upstream fish migration will be monitored at a portion of the river 1.5 mi upstream from the
bridge project site (see Figure 2), where the river narrows and deepens into a choke point. There
we can ensure complete coverage of the river with an echosounder acoustic beam from bank to
bank, and count migrating fishes as they pass. All migrating fish will pass first by the
construction site (Site 1) then through this location (Site 2).
6
Figure 1. Tar River channel in the center, Kennedy Creek in upper center and the City of Washington, NC is in
the upper right. The location of the Route 17 by-bass bridge crossing and pile driving activity (days 1-6) (Site 1,
inset photo) is shown. The BioSonics echosounder monitoring stations will be located at the existing US Route 17
bridge downstream and at a site where the Tar River narrows upstream (Site 2), shown on the western end of this
image. Caging studies with adult and larval fishes will be done at place marks (for pile driving event 1), then will
follow across the river in successive weeks.
A continuously operating long-term hydroacoustic monitoring stations downstream at the
existing Route 17 bridge in the deepest part of the channel (4 m) at Site 2 (6.5 m), using a
BioSonics scientific echosounder and two transducers aimed at the center of the Tar River. Each
fish (this includes all species that pass by site, which will be compared with data collected in gill
nets, see Job 3) passing by the downstream (where a portion of the river channel can be covered)
and upstream Site 2 (complete coverage of the river) will create a target on an echogram as the
sound energy bounces off them and is received by the echosounder transducer. These fish targets
will be counted using standard echo-sounder analysis methods (BioSonics Visual Analyzer will
be used to prepare echograms and generate reports). Use of 420 kHz split-beam echosounders
will allow scientifically defensible hydroacoustic data to be collected continuously (day and
night) for entire study period March 1 - June I (or longer if construction is prolonged). The 420
kHz acoustic pings are above the hearing range of all fishes in the river, including American
shad, thus it should not cause any avoidance or prevent fish passage. Information on fishes will
be collected on target strength (proportional to fish size), direction of travel (upstream or
downstream) and velocity of target (m/s). We hypothesize that during periods of pile-driving and
heavy construction activity at the bridge downstream, there will be fish avoidance and migration
will be impeded. Fish will remain below the construction site (near the Washington Route 17
Bridge) until the activity associated with pile-driving ceases, and then they will migrate upstream
again past Site 2 (right to left in the image in Figure 1).
We will use two BioSonics 420-kHz split-beam transducers, with swivel mounts for aiming the
beam, a 600-foot waterproof cable, and a rack-mount low power solid state computer for data
logging with RAID hard drive, (above equipment will be leased from BioSonics, Inc. for 4
months), a DTX surface unit (ECU-owned) (Figure 2), 2 TB back-up hard drives (to be
purchased), power supply (12-V battery bank to be purchased), and an environmental housing
(elevated lockable shed, to be purchased). We will place the environmental housing on the north
side of the river bank, and string the cable across the river (weighted down with chain). On each
side of the Tar River, we will position one of the 420-kHz transducers, aimed in such a way as to
direct the echosounder acoustic beam in the entire water column in the river (Figure 3). This
installation will take 3-4 days and will be done by ECU divers, ECU boat operators, and
supervised by BioSonics scientists. BioSonics is experienced in river hydroacoustic studies of
fish passage and counting. BioSonics will be responsible for preparation and shipping of
equipment, installation, data storage and analysis, report preparation.
Figure 2. BioSonics DTX digital scientific echosounder, with laptop and 420-kHz transducer. Rack-mounted
low power computer for data collection and real-time analysis. Laptop computer will not be required for a fixed-site
deployment at Site 2.
8

--------------;---------;---------------- _---
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2, obtained on 13 Feb 2009. Lower panel is a schematic from other BioSonics fish passage studies down elsewhere
for illustration. We will use 420-kHz transducers, which will minimize any avoidance responses of the migrating
clupeid fishes. On the Tar River at Site 2, an environmental housing will be built and a multiplexing cable (600'
long) will be laid across the river bed, connecting the transducer to the DTX echosounder in the housing. A similar
set up will be deployed downstream at the existing Washington Route 17 bridge. The housing at each site will be
elevated 10 feet to allow for flooding and will enclose a rack-mounted computer and RAID hard drive to store the
echosounder data.
Experimental Design
When the pile driving is operational, we expect the fish counts to decline and when the operation
ceases, we expect the fish counts to be increased. We also expect fish counts at Site 2 to
increase as anadromous species continue their spawning runs (pre-spawn, spawning, and post-
spawn) April through June 2009. We will sample continuously to estimate variability in the fish
count data. We will compare the fish counts obtained when pile driving operations are underway
(acoustic disturbance period, a cumulative amount of 4 hours per day, 1 day per week) to those
obtained during non-pile driving days (a comparable period of time during the preceding week
and following week (control periods). BioSonics scientists can examine up to 12 hours of data
the day after an event of pile driving (comparing before, during and after periods) to see if there
was a decline in the number of fish heading upstream. We expect that the cumulative number of
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9
fish passing at the echosounder upstream location will fall below the 95% confidence interval
around the mean fish counts obtained during control periods.
Job 3 - Adult Fish Identification and Relative Abundance during Upstream Passage and
Mortality Survey (Rulifson)
The purpose of Job 3 is to identify the species of fish observed on the echosounders during Job 2,
and to determine relative abundance of those fishes upstream, within, and below the construction
site on the day prior to, during, and one day after pile driving events. Also, we will survey the
river's surface waters for any mortality of fishes associated with the pile-driving events.
Shallower portions of the river cannot be sampled with electroshocking units, so therefore only
gillnets will be used for several reasons. First, electroshocking cannot be conducted in the area
near the echosounders or near the hydrophones. The manufacturers are concerned about
electronic failure of these devices if high electrical current passes through the water in which
they are deployed. Second, no brackish water electroshocking unit is available with such late
notice.
Gillnet Sampling Design
Gillnets will be set in the deep reaches of the river downstream, at, and above the
construction site one day prior, during, and one day after the event. Site 1 is the new bridge
construction site. Site 2 is the upstream constricted area where the echosounders will be located
(Figure 1). Site 3 is near the existing U.S. Highway 17 bridge and the echosounders located
there, as described in Job 2. Gillnets will be set for 30 minutes or 1 hour (as determined by
abundance) in the south shallow water area (< 2 m), in the river mid-channel (2-8 m), and north
shallow water area of the river at Sites 1, 2 and 3. Samples will be taken in the morning, again in
the afternoon, and at sunset. We will set gillnets in the deep areas of the river because this is the
area being monitored by the echosounding units, and we want to measure the same portion of the
river. Surface floating gillnets will be used with anchors, constructed of 104 (.33 mm) twine, to
ensure that any fish in the area are caught. There will be 126-yards of gill nets (three 42-yard
nets) in each depth section of the sample areas: I to 5 inch stretch mesh, %2 inch stretch mesh
intervals. Fish will be enumerated by species and returned to the water. Key species (striped
bass, hickory shad, American shad, and river herring) may be retained for use in Job 4 (see
below). Standard water quality parameters will be measured during the study including
dissolved oxygen (mg/L) and percent saturation, water temperature (°C), and salinity (ppt).
Mortality Survey
In addition, we will conduct a visual examination of shallow and shoreline areas for
injured and dying/dead fish. We will use the protocol and methods that are generally consistent
with the NC DENR methods outlined in their fish kill surveys (based on the American Fisheries
Society publication, Investigation and Valuation of Fish Kills, Special Publication 24 (1992).
The protocol for this mortality survey is as follows:
1. We will survey the river from the upstream constriction (Site 2) to the existing NC 17 bridge
(Site 3). We will conduct river surveys (back and forth across the river using ten belt transects of
20-m width) from a slowly moving boat with two lookouts, one looking to each side within 10 in
10
of the boat. We will conduct these in the morning and again late afternoon one day prior to pile-
driving (control), on the day of pile driving, and one day after the event. No nighttime surveys
will be conducted due to dangers of navigating the shallows at night, and the inability to see
floating fish in the dark.
2. All fish encountered floating at the surface during the survey will be retrieved, identified to
species, and examined externally for damage. These will be noted on field data sheets.
3. Any fish collected will be bagged, labeled with capture location coordinates and time of
collection, and returned to the lab for post-mortem analysis to determine cause of death.
Methods for this step are detailed in the next section (Job 4 Experimental Adult Mortality Study).
4. Standard water quality parameters will be measured at each location of a collected dead or
injured fish.
Job 4 - Experimental Injury and Mortality Study of Selected Fish Species (Overton,
Harris, Krahforst and Sprague)
The purpose of Job 4 is to test examine the effects of sound on short-term mortality on sub-adult
and adult fish.
Experimental Design:
We will measure the physical effects of the sound produced by pile-driving on fishes. Pile-
driving causes a propagation of sound outward from the source through both the water column
and the sediment, causing an increase in both sound pressure and particle motion. This leads to
an increase in fluid motion which can impact the fluid-filled and air-filled structures within a
fish, causing tissue damage (e.g., ruptured swim bladder), blood embolisms, neurotrauma, and
even death (Hastings and Popper 2005). Some of these damaging effects may not be observed
immediately but may be more prominent 24h or more after the pile-driving event (McCauley et
al. 2003).
In order to measure the physical effects of pile-driving on fish, we will obtain threadfin shad
(Dorosoma petenense), striped bass (Moron saxatilis), or white perch (Moron americanus)
from hatcheries and fish farms. Bennett Wynne of the NC WRC has offered to provide threadfin
shad from their hatcheries. These species were selected because the threadfin shad (Order:
Clupeiformes) is a hearing specialist and the striped bass and white perch (Order: Perciformes)
are hearing generalists. Hearing specialists have a direct connection between the swim bladder
and the otolith sac (contains the hearing bone) and are generally thought to be more sensitive to
sound propagation (Hastings and Popper 2005) and therefore may exhibit increased signs of
stress during a pile-driving event. If possible, we will try to gather additional species including
Alosa sp. and striped bass from gillnetting, rod and reel, and seining for at least one experiment.
These are fragile species, and it is unknown about the extent of damage that will result from
capture and transport.
Fish cages will be constructed of aluminum frames with knotless nylon netting and submersed at
each location. We may consider two plastic fish baskets tie-wrapped together (these are orange
baskets used to hold commercial fish catches) as an alternative cage design, if we can
demonstrate that they have small acoustic artifacts and they are cost-effective. Five fish of each
II
species will be placed into separate cages or baskets, and there will be two cages of each species
placed at 10 in, 50 in and 400m (control) from the pile driving site. We will also conduct a
fourth treatment of fishes that are handled in a similar manner to all treated fish, but never
exposed to sound effects of pile driving (Handling Stress Control, to be held in separate cages in
Tranter's Creek). This makes a total of 12 cages per 20-minute pile driving event. A hydrophone
with digital recorder will be placed adjacent to the cages at 10 in, 50 in and 400 in to measure
sound exposure levels. Each cage will be suspended in the water column for a pile-driving event
at Bents 91, 92, and 93 (these have water depths >2 m). The holding cages in Tranter's Creek
will be at 2 in. Fish will be moved to the experimental site and re-acclimated to depth of 2 in
(we will allow -40 min for fish to adapt to the hydrostatic changes, based on a change in
pressure of 0.18 atm for 0-2 in depth change and the swim bladder adaptation equation for cod in
Harden-Jones and Scholes 1985). Standard water quality parameters will be measured during the
study including dissolved oxygen (mg/L) and percent saturation, water temperature (°C), and
salinity (ppt). After the 20-minute pile driving event the cage will be checked, dead fish will be
removed and stored in a bag filled with water, then placed on ice for storage and later necropsy.
Any remaining fish will be separated into a group for immediate necropsy (see euthanization and
necropsy procedure below) and a delayed mortality group, which will be held alive for continued
observation. We will take blood samples from the immediate necropsy group using a caudal
puncture. Delayed mortality individuals will be placed into cages in Tranter's Creek and held for
up to four days. We expect that with 12 pile driving events with each Bent location per day, we
can obtain up to three replicated tests per day, with cage locations adjusted as the construction
work proceeds. We plan to conduct these experiments beginning April and May. We will not
conduct experimental caging studies in shallow water driving sites on the south side of the river
(Bents 90 - 84), because the sound propagation in shallow water sites (below 2 m) will be above
the frequency heard by most fishes. By this time fish mortality rates will be established, and
fishes will be largely past the mid-channel locations. This Job 4 will coordinate very closely with
the schedule of Job 1. Larval mortality studies (Job 6) will begin then in shallow waters.
Necropsy studies and delayed mortality studies will be carried out in the 5 days following each
experimental exposure study. Five fish of each species will be sacrificed immediately after
exposure for necropsy. The remaining fish from each treatment group will be placed in delayed
mortality cages (one per species and treatment) and monitored for survival. Any dead or injured
fishes will be examined at four-hour intervals for 96 hr. Necropsies on all fishes will include
examination for hemorrhaging in tissues and eyes, swim bladder ruptures, and gross examination
of tissues. To assess the possibility of neurotrauma, we will dissect out the otolith sacs and take
scanning electron microscope images of the sensory epithelium (macula) to ascertain if there is
any impact on the ciliary bundles (used for hearing) due to the pile-driving sounds.
Blood serum chemistry will be used as a measure of short-term stress. Blood will be immediately
collected from five fish from each treatment following removal from experimental cages with a
caudal puncture using a 3-ml plastic syringe and a 22-gauge needle. The blood sample will be
transferred gently to a plastic vial without anticoagulant and allowed to clot at air temperature.
The fish will then be weighed and measured, then placed on ice for further evaluation. Serum
will be separated by centrifugation at 14,000x g for 3 minutes. Plasma will be immediately
removed, and then frozen at -40°C until analysis. Serum samples will be sent to Antech
Laboratories,_ Farmingdale, New York for determination of the blood constituents.
12
Statistical analysis will include ANOVA and non-parametric tests (Kruskal-Wallis) as
appropriate for each response variable (% mortality, injured, numbers with various tissue
damaged). Mean and variance of each response variable will be assessed statistically with respect
to two factors: distance from pile-driving site (10m, 50m, 400m, and handling stress control
treatments) and hearing capabilities of each species (hearing generalist vs. hearing specialist).
We will use water depth and Sound Exposure Levels (SEL) measured at each distance as a
covariates in the analysis.
Job 5 -Larval Fish Downstream Migration (Rulifson)
The presence of larval fish moving through the construction site region of the river could result
in extensive mortality of larvae (Job 6). To determine presence and absence of larval key fish
species, and relative abundance, we will conduct larval fish sampling using push-nets and a pair
of ichthyoplankton nets pulled in an oblique manner through the water column. The push-nets
(1-m wide by 0.5-m deep and constructed of 1-mm nitex mesh) will be mounted on a metal skiff
and pushed through the water to obtain samples of larval Alosa, which tend to be near the surface
during certain times of the day. Push-nets will be used for 5 minutes in the north and south side
of the river shallow areas and in mid-channel at Sites 1, 2 and 3. Samples will be taken in the
across the river from North to South. Caging studies will follow the pile driving progress, with distance from the
pile driving sites 1-6 (10 m, and >100m, flowing along river depth contours upstream). Distance from each pile
driving site will be measured at the start of each test using a laser rangefinder.
13
morning and again in the afternoon: Paired oblique plankton net also will be used in the mid-
channel locations at Sites 1, 2 and 3 in the morning and afternoon. The paired plankton net will
be towed in an oblique manner through the water column for 5 minutes. Samples will be
preserved in formalin and returned to the lab for processing. Standard water quality parameters
will be measured during the study including dissolved oxygen (mg/L) and percent saturation,
water temperature CC), and salinity (ppt).
Thus, all ichthyoplankton sampling will be on the day before pile-driving, on the day of pile-
driving, and day after pile-driving events at Sites 1, 2, and 3. Samples will be placed in 5%
formalin containing Rose Bengal dye and returned to the laboratory for processing. Larvae will
be counted by species and reported as number of larvae per cubic meter. In the laboratory, we
will look for dead fish (% of fish occurring at the time of collection) in the samples by examining
for partial decomposition.
Job 6 Larval Fish Injury and Mortality (Overton and Sprague).
The purpose of Job 6 is to test examine the effects of sound on short-term mortality and
physiological condition of larval American shad and striped bass.
Experimental Design:
Experimental Barges will be constructed to each hold six experimental tanks (Figure 5). A
single barge will be anchored 10 m upstream away from pile driving activities and second
(control)will be located at least 100m from the bridge construction in an area not exposed to the
sound produced by the pile driving activities. Hydrophones will be anchored at each barge to
estimate the cumulative sound exposure. Larvae (-5-25 days old) will be obtained from state
hatcheries and held in ECU rearing facilities until the start of the pile driving event.
Approximately 10-15 minutes before a pile driving event, larvae will be placed into the
experimental barge tanks (Density of larvae not to exceed 15 individuals/m3 for a single pile
driving event. The duration that experimental fish are exposed to sound will depend on
operational pile driving activities. Also the number of trials conducted for each species will
depend on the availability of larvae. We are expecting two to three days of testing for each
species. Standard water quality parameters will be measured during the study including dissolved
oxygen (mg/L) and percent saturation, water temperature (°C), and salinity (ppt).
Immediately, after each pile driving event (-20 minutes per event), dead and alive fish will be
counted to estimate acute mortality. This job will coordinate very closely with the schedule of
Jobs 1 and 5.
Larval Histology and Acute Stress Test:
Histology: Test fish (Dead and alive) will be randomly selected from each tank stored and
preserved in Histopaks. The histological samples will be sent to the USGS Leetown Science
Center (Keameysville, WV-Dr. Vicki Blazer) Fish Health Laboratory for processing. The larvae
will be stained with haematoxylin, eosin, and alcian blue (pH 2.6) sectioned whole for general
gross observation.
14
The remaining live fish will be subjected to acute salinity test (SST) in an onshore mobile
laboratory. The salinity stress test is widely used to estimate the quality of post-larvae (PL) and
as a predictive indicator of performance; on the assumption that it will predict future
performance. Here, we will use a high-salinity stress test (40 ppt, @ ambient temperatures °C) on
larvae to test the cumulative effects of sound exposure. Larvae from each replicate will be
transferred carefully with a wide glass pipette to each of two small glass beakers (25 larvae per
beaker), which contained 50 ml of filtered water. Larvae in one beaker will be exposed to 40 ppt
salinity by adding 50 ml of high-salinity water (Instant Ocean®), which will result in 100 ml
test-solution of 40 ppt in each beaker. Larvae in the other beaker from each replicate served as
control and 50 ml of normal sea water (20 ppt) was added by the start of the test. The salinity
will be checked by a hand refractometer. Mortality will be recorded every 10 min until all the
larvae in the exposed groups are dead. The larvae were considered dead if they were opaque,
their body was bent and they did not respond to a gentle touch of the tail. The treatment effects
will be compared using a two-way ANOVA.
construction.
Larval Histology and Acute Stress Test:
Figure 5. Proposed experimental design for larval fish exposure trials Rt 17 bridge
15
Histology: Test fish (Dead and alive) will be randomly selected from each tank stored and
preserved in Histopaks. The histological samples will be sent to the USGS Leetown Science
Center (Kcameysville, WV-Dr. Vicki Blazer) Fish Health Laboratory for processing. The larvae
will be stained with haematoxylin, eosin, and alcian blue (pH 2.6) sectioned whole for general
gross observation.
The remaining live fish will be subjected to acute salinity test (SST) in an onshore mobile
laboratory The salinity stress test is widely used to estimate the quality of post-larvae (PL) and as
a predictive indicator of performance; on the assumption that it will predict future performance.
Here we will use a high-salinity stress test (40 ppt, @ ambient temperatures °C) on larvae to test
the cumulative effects of sound exposure. Larvae from each replicate will be transferred
carefully with a wide glass pipette to each of two small glass beakers (25 larvae per beaker),
which contained 50 ml of filtered water. Larvae in one beaker will be exposed to 40 ppt salinity
by adding 50 ml of high-salinity water (NaCl dissolved in filtered sea water), which will result in
100 ml test-solution of 40 ppt in each beaker. Larvae in the other beaker from each replicate
served as control and 50 ml of normal sea water (20 ppt) was added by the start of the test. The
salinity will be checked by a hand refractometer. Mortality will be recorded every 10 min until
all the larvae in the exposed groups are dead. The larvae were considered dead if they were
opaque, their body was bent and they did not respond to a gentle touch of the tail. The treatment
effects will be compared using a two-way ANOVA.
REPORTING
Event Reports
Following each pile driving event, an Event Report will be prepared for the agencies
summarizing the results of the various research activities during that event (see Timeline). For
example, Event 3 might include results of the hydrophone survey (Job 1), adult fish acoustic
survey (Job 2), adult fish ID and abundance study (Job 3), adult injury and mortality study (Job
4), and larval fish assessment (Job 5). There may be no Job 6 report because the larvae may not
be available for sound exposure until later in the study.
Final Report
A Final Report will be prepared detailing the methodologies and results of all experiments and
field sampling conducted during the study. Due to the amount of data collected, it is anticipated
that the final report will require considerable time requirement. We anticipate that a draft of the
report can be sent to agencies for review by Feb 14, 2010.
REFERENCES CITED
Harden Jones, F. R., and Scholes, P. 1985. Gas secretion and resorption in the swimbladder of
the cod Gadus morhua. Journal of Comparative Physiology B, 155: 319-331.
Hastings, M. C.; and A. Popper. 2005. Effects of Sound on Fish. Final Report #CA05-0537.
Project P476 Noise Thresholds for Endangered Fish. California Department of
16
Transportation Division of Research and Innovation and Division of Environmental
Analysis. Sacramento, CA.
McCauley, R. D; J. Fawrewll; and A. N. Popper. 2003. High intensity anthropogenic sound
damages fish ears. J. Acoust. Soc. Am. 113: 638-642.
17
BUDGET
Salaries $110,732
Fringe Benefits (1800) $16,863
Total $127,595
Other Contracted Services (1900) $194,208
Consumable Supplies/Matedals (2000) $19,400
Travel/Subsistence(3100) $10,428
Other Direct Costs/Services
Communications (3200) $250
Other Direct Costs -- CurrentServices (3000) $3,850
Other Direct Costs - Fixed Charges (4000) $5,000
Other Direct Costs - Specify $14,550
Total $23,650
Equipment Purchase (5000)
$50044,9991Item $0
Over$5,000/Item $0
Total $0
Sub-Contracts (1995) $7,000
(MTDC) Modified Total Direct Costs: $382,281
(F & A)Facilities & Administrative Costs (4800): $99,394
Total Budget $481,675
18
BUDGET NOTES
SALARIES

Name Base
Salary #
Mos. %
Effort Sponsor
Salary Benefits
Summer Salary/9-Month Faculty (1000)
Roger Rulifson $89,464 2.0 100% $19,881 $3,799
Anthony Overton $63,894 2.0 100% $14,199 $2,713
Joseph Luczkovich $71,750 2.0 100% $15,944 $3,047
Patrick Harris $35,865 2.0 100% $7,970 $1,523
Mark Sprague $68,571 2.0 100% $15,238 $2,912
Subtotal $73,232 $13,994
Student Pay/Academic Year (1000)
Cecila Krahforst $10,800 4.0 100% $4,800 $367
Subtotal $4,800 $367
Hourly Personnel (1000)
Boat Operator 1 $15.00 300 $4,500 $344
Boat Operator 2 $15.00 300 $4,500 $344
Sound Recorder 1 $15.00 300 $4,500 $344
Sound Recorder2 $15.00 300 $4,500 $344
Diver 1 $15.00 40 $600 $46
Divert $15.00 40 $600 $46
Unnamed undergrad-field $15.00 500 $7,500 $575
Unnamed undergrad-lab $10.00 600 $6,000 $459
Subtotal $32,700 $2,502
Total $110,732 $16,863
Total $127,595
OTHER COSTS
Other Contracted Service (1900)
Fixed Site Hydroacoustic monitoring at two sites as per agreement with Biosonics, Inc. $191,008
Antec, Inc $3,200
Total $194,208
Consumable Supplies/Materials (2000)
Laser Rangefinders 2 @ $250 $500
Edirol R-09HR Digital Recorders (3 @ 450) $1,350
Fish purchases from Aquaculture farms $2,000
8 GB SDHC Flash Memory Cards (9 @ $50) $450
Handheld GPS (2 @ 350) $700
External Hard Drive (4 TB of disk space) $800
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