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NOAA TR NMFS SSRF-677
DIVig E
SION OF FISHES
Us RE Ary om
a, NATION: Muse
NOAA Technical Report NMFS SSRF- 6/7
A UNITED STATES
DEPARTMENT OF
COMMERCE
PUBLICATION
U.S. DEPARTMENT OF COMMERCE
National Oceanic and Atmospheric Administration
National Marine Fisheries Service
Abundance of
Benthic Macroinvertebrates in
Natural and Altered Estuarine Areas
GIL GILMORE and LEE TRENT
SEATTLE, WA
April 1974
NOAA TECHNICAL REPORTS
National Marine Fisheries Service, Special Scientific Report—Fisheries Series
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620. The Trade Wind Zone Oceanography Pilot Study. Part IX: The sea-level wind field
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621. Predation by sculpins on fall chinook salmon, Oncorhynchus tshawytscha, fry of
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622. Number and lengths, by season, of fishes caught with an otter trawl near Woods
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623. Apparent abundance, distribution, and migrations of albacore, Thunnus alalunga,
on the North Pacific longline grounds. By Brian J. Rothschild and Marian Y. Y. Yong.
September 1970, v + 37 pp., 19 figs., 5 tables.
624. Influence of mechanical processing on the quality and yield of bay scallop meats. By
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625. ‘Distribution of salmon and related oceanographic features in the North Pacific
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626. Commercial fishery and biology of the freshwater shrimp, Macrobrachium, in the
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1971, iii + 7 pp., 1 fig., 3 tables.
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5 tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
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638. Length-weight relations of haddock from commercial landings in New England,
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figs., 6 tables, 10 appendix A tables. For sale by the Superintendent of Documents, U.S.
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639. A hydrographic survey of the Galveston Bay system, Texas 1963-66. By E. J. Pullen,
W. L. Trent, and G. B. Adams. October 1971, v + 13 pp., 15 figs., 12 tables. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
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640. Annotated bibliography on the fishing industry and biology of the blue crab,
Callinectes sapidus. By Marlin E. Tagatz and Ann Bowman Hall. August 1971, 94 pp. For
sale by the Superintendent of Documents, U.S. Government Printing Office, Washington,
D.C. 20402.
641. Use of threadfin shad, Dorosoma petenense, as live bait during experimental pole-
and-line fishing for skipjack tuna, Katsuwonus pelamis, in Hawaii. By Robert T. B.
Iversen. August 1971, iii + 10 pp., 3 figs., 7 tables. For sale by the Superintendent of
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642. Atlantic menhaden Brevoortia tyrannus resource and fishery—analysis of decline.
By Kenneth A. Henry. August 1971, v + 32 pp., 40 figs., 5 appendix figs., 3 tables, 2
appendix tables. For sale by the Superintendent of Documents, U.S. Government Printing
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644. Inhibition of flesh browning and skin color fading in frozen fillets of yelloweye
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Continued on inside back cover.
U.S. DEPARTMENT OF COMMERCE
Frederick B. Dent, Secretary
NATIONAL OCEANIC AND ATMOSPHERIC ADMINISTRATION
Robert M. White, Administrator
NATIONAL MARINE FISHERIES SERVICE
NOAA Technical Report NMFS SSRF-677
Abundance of
Benthic Macroinvertebrates in
Natural and Altered Estuarine Areas
GIL GILMORE and LEE TRENT
yOUTOn,
& %
= Bs
3 Z Seattle, WA
z BD April 1974
2 =
vs ~~
776 -491° WEE MaDe of Documents, U.S, Government Printing Office
The National Marine Fisheries Service (NMFS) does not approve, rec-
ommend or endorse any proprietary product or proprietary material
mentioned in this publication. No reference shall be made to NMFS, or
to this publication furnished by NMFS, in any advertising or sales pro-
motion which would indicate or imply that NMFS approves, recommends
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herein, or which has as its purpose an intent to cause directly or indirectly
the advertised product to be used or purchased because of this NMFS
publication.
Introduction
Study area and methods
Station description and environmental data
Relative abundance
Comparison between canal, marsh, and bay
Discussion
a
i)
= So 0>
CONTENTS
Figures
Study area and sampling locations in the Jamaica Beach area of West Bay, Texas ..........
Sand, silt, and clay fractions of the sediments by station and zone ...................-005-
Average volumes of organic matter (cord grass roots, submerged marine grasses, and organic
detritus combined) by station and zone, March-October 1969 ............. 0.0000 ee ee eee
Average volume of organic matter by date, station, and zone ................. eee eeeeeeeee
Average dissolved oxygen by date for all six stations combined and for the stations having
theshishesiwandulowestgaveracenvalllc summer tars eaceistcic errs kicieiskec iin ocfeectere esters oii
Average numbers and volumes (stations combined) of organisms caught per sample by
taxonomic.eroupiandy zone) March-October O69 i lies yarcisverapst dei ree casieveheve) #1 ais. snersie eis ensauele
Average number of polychaetes caught per sample by date, station, and zone ..............
Average number of crustaceans caught per sample by date, station, and zone ..............
Average number of pelecypods caught per sample by date, station, and zone ...............
Average number of nemerteans caught per sample by date, station, and zone...............
Average numbers and volumes of benthic macroinvertebrates caught per sample by area,
taxonomic group, and for the groups combined, March-October 1969 ................---.
Tables
List of taxonomic groups and total numbers of specimens collected by station and zone .....
Comparisons between zones by taxon and station of the mean numbers of organisms col-
lectedk(pained=comparisoni/-test) nner eer ORO EGC GORE CR ne toner
Comparison between stations by taxon and zone of the mean numbers of organisms collected
(woowavaeanaly sis) Ofsvallance) iy rer rene ceteris Sey alewe layin shah unshared), ac a ance ae
Appendix Tables
Numbers of animals caught by station, date, taxon, and volume of two samples in shore
AYU SASH OO OODOPOD OD DONO DOOUDDOOOOU DU OO IUOOOUC OOOOH OOOO pOOD OO DOD ODO ODDO OOO OUDU Oto
Numbers of animals caught by station, date, taxon, and volume of two samples in
COTIAE GEARS BIVENS or Gro. DAREN CONC Ore Oe NEE ORCL Sr CORT ee DRE CE REL Cs CME OEIC Gea tetans Ory ata
ill
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NINANN OD WwW won fpNNe
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in
Abundance of Benthic Macroinvertebrates in Natural and
Altered Estuarine Areas!
GIL GILMORE? and LEE TRENT?
ABSTRACT
The abundance of benthic macroinvertebrates during March-October 1969 in West Bay, Texas, was
compared between 1) a natural marsh area, 2) an adjacent marsh area altered by channelization, bulk-
heading, and filling, and 3) an open bay area. Animals representing four phyla were caught. Abundance
indices (areas combined) of the four groups in terms of numbers were 66.4% polychaetes, 29.6% crusta-
ceans, 2.5% pelecypods, and 1.5% nemerteans; volumes were 44.0% polychaetes, 40.8% pelecypods,
10.7% nemerteans, and 4.4% crustaceans.
When all organisms were combined, they were slightly more abundant numerically and over twice as
abundant volumetrically in the marsh than in the canals and were least abundant in the bay. Polychaetes
were most abundant in the canals and least abundant in the bay; abundance was highest at stations with
low to intermediate amounts of silt and clay or where vegetative matter was composed mostly of live sea
grasses or detritus. Crustaceans were more abundant in the natural marsh than in the other two areas and
showed a definite preference for sandy substrate in marsh areas. Pelecypods were numerically most
abundant in the bay but volumetrically the marsh had the highest standing crop. Nemerteans were most
abundant in the marsh and least abundant in the bay.
In general, the seasonal abundance of polychaetes and nemerteans varied little during the study,
whereas crustaceans and pelecypods were abundant only during the spring and early summer. An
exception to this seasonal abundance pattern was the reduction in numbers of polychaetes at the upper-
most canal station where the habitat was apparently unsuitable due to low oxygen levels during the
summer and early fall.
INTRODUCTION
Development of bayshore property into housing
sites by dredging, bulkheading, and filling is occurring
in many estuaries. When this property is developed,
shallow bay and tidal marsh areas are dredged out or
filled with spoil, thus changing the environment for
marine organisms. The effects of the resulting en-
vironmental changes on the abundance of benthic or-
ganisms are poorly understood.
Some studies on the succession and abundance of
benthic marine animals in canals following excavation
indicate that succession may occur rapidly and that
climax communities may be established within 2 yr
(Brandt, 1897; Reish, 1956, 1957). For example, peaks
in the number of species and specimens of benthic
invertebrates were reached about 2 yr after seawater
entered a newly constructed boat harbor in California,
and the abundance and species diversity in the harbor
* Contribution No. 335, Galveston Laboratory, Gulf Coastal
Fisheries Center, National Marine Fisheries Service, Galveston,
Texas.
* Marine Laboratory, Texas A&M University, Galveston, TX
77550.
® Panama City Laboratory, Gulf Coastal Fisheries Center, Na-
tional Marine Fisheries Service, NOAA, Panama City, FL 32401.
2 yr after opening were comparable to those in adja-
cent natural areas (Reish, 1961).
In other cases, however, climax communities may
be altered or natural succession may not occur. Reish
(1961) found that the benthic population in a boat har-
bor in California decreased markedly during the third
year, probably because of low dissolved oxygen.
Taylor and Saloman (1968) concluded that soft de-
posits in the canals of a Boca Ciega Bay housing de-
velopment in Florida were in some way unsuitable for
most benthic invertebrates that were found in the
natural areas of the same bay. After 10 yr, recoloniza-
tion of canal sediments by benthic organisms was neg-
ligible, and it appeared doubtful that soft sediments of
the canals would ever support a rich or diverse in-
fauna. The habitat in Boca Ciega Bay was, however.
changed drastically by channelization of the natural
bay area which was sandy and shallow, and had con-
tained numerous turtle grass (Thalassia testudinum)
beds and oyster reefs.
The objectives of our study were 1) to determine the
relative abundance of benthic macroinvertebrates in
three habitats: a natural marsh characterized by cord
grass (Spartina alterniflora) and by sparse, submerged
vegetation; a previously similar area altered by dredg-
ing, bulkheading, and filling; and an open bay area;
and 2) to relate invertebrate abundance within each
area to sediment particle size, amount of plant matter,
and dissolved oxygen.
STUDY AREA AND METHODS
The study area in West Bay, Texas, a part of the
Galveston Bay System (Fig. 1), included a natural
marsh area, an open bay area, and an area that was
similar to the natural marsh prior to alteration by
channelization, bulkheading, and filling for the
Jamaica Beach housing development. The developed
area, which included about 45 hectares of emergent
marsh vegetation, intertidal mud flats, and subtidal
water area prior to alterations was reduced to about 32
hectares of subtidal water area by dredging and filling
between 1958 and 1960. The water volume (mean low
tide level) was increased from about 184,000 to about
394,000 cubic meters.
Six sampling stations—two in the canals of the al-
tered area, three in the natural marsh, and one in the
open bay—were established (Fig.1). The stations were
numbered nonconsecutively and correspond to those
reported by Trent, Pullen, and Moore (1972). Sam-
pling was conducted in two zones, ‘‘shore’’ and
“center,” at each station except at the open bay sta-
tion. In the altered area, shore samples were taken on
each side of the canal at each station (1 and 4) 1 m
away from each bulkhead; center samples were taken
near the center of the canal along a transect perpen-
dicular to the bulkheads. Replicate samples were
taken from the center at each station, so that two sam-
ples were available from each zone for each sampling
date; the average of the two samples was used as the
observation in the statistical tests. Samples similar to
those from the canal area were taken at stations 6, 7,
and 8 in the intertidal zone adjacent to the cord grass
and near the center of the bayou or lake along tran-
sects perpendicular to the shoreline. Replicate sam-
ples were taken at station 10 in the open bay.
Bottom samples were taken at 14-day intervals from
25 March to 21 October 1969, with a metal cylinder 14
cm long and 9.6 cm in diameter. To obtain a sample
the cylinder was pressed about 11 cm into the bottom
sediments, capped on each end with plastic lids, and
brought to the water’s surface. Each sample contained
about 800 cm® of bottom materials and represented a
surface area of 1/138 m?.
The samples were refrigerated within 2 hr after col-
lection. The following day, each sample was emptied
into a sieve having a mesh size of 420 um, and the
material was washed until the fine sediments passed
through the sieve. The remaining material, including
macroinvertebrates, was stored in a 10% Formalin
solution.
Macroinvertebrates and plant material in the pre-
served samples were separated from the shell and
sand. Animal volume for each pair of samples was
determined to the nearest 0.1 ml by displacement in a
graduated 10-ml centrifuge tube containing a previ-
ously recorded volume of water. The volume of each
phylum taken at each station and zone was determined
at the end of the study by combining the individuals of
a phylum from the 32 samples. The volume of plant
material was determined to the nearest milliliter in a
30-ml graduated cylinder.
The animals were separated and identified, usually
to family, and the number of individuals in each group
was recorded for each pair of samples. As suggested
by Holme (1964), only whole animals or portions of
animals containing the anterior end were counted to
avoid recounting the same animal.
The dissolved oxygen content of the water, taken 15
cm above the bottom at the center habitat of each
station, was measured using a modified Winkler
method. The water samples were taken about midday
and midnight during the same 24-hr period that the
bottom samples were collected.
Samples for sediment analyses were taken at each
station and zone on 12 August. Particle-size composi-
tions (percents by weight) were determined using a
series of sieves and soil hydrometers. Ranges in parti-
cle size were: sand, 2.0-0.62 mm; silt, 0.061-0.004 mm;
clay, 0.0039-0.001 mm.
STATION DESCRIPTION AND
ENVIRONMENTAL DATA
Sampling stations were located at various distances
from the West Bay shoreline (Fig. 1). Water depths
(mean low tide level) in the shore zone ranged from 0.0
to 0.6 m and in the center zone from 0.2 to 1.6 m.
The percentage compositions of bottom sediments
varied considerably between stations and between
zones (Fig. 2). In the canals, silt and clay components
were more abundant in the center than in the shore
zone. In the marsh, compositions of sediments were
similar between zones at stations 7 and 8, but at station
6 a higher percentage of silt and clay occurred in the
center than in the shore zone.
The plant material taken in the samples was com-
posed of live cord grass roots, attached marine grasses
(mostly Diplanthera wrightii), algae (mostly
Ectocarpus sp.), and detritus. In general, all organic
matter collected at the canal stations (both zones) and
in the open bay consisted of detritus and small
amounts of attached algae. Cord grass roots were
dominant in the samples from the shore of the marsh,
whereas in the center, detritus was usually dominant,
although attached grasses and algae were also present.
The volume of plant material was many times great-
er in the shore zone (stations 6, 7, and 8) of the marsh
than in either zone within the canals (Fig. 3). Most of
this vegetation, however, was not decomposed; thus,
detrital and filter feeders could not utilize it.
Differences in the volume of plant material in the
center zones of all stations indicated that a major
source of plant material in the canals originated from
an outside source. The lowest volume of plant material
GALVESTON BAY SYSTEM
Figure 1.—Study “area and
sampling locations in the Jamaica
Beach area of West Bay, Texas.
CANAL
leak
60F _ it
40-+
MARSH BAY
PERCENT
SHORE
4of
CENTER
PERCENT
80F
100 A 6 .
STATION
Figure 2.—Sand, silt, and clay fractions of the sediments by station
and zone.
occurred at station 1, the farthest station from the
open bay. Plant volume at station 4 (close to the bay)
was similar to the volumes at stations 6 and 7 in the
marsh. The highest volume of plant material occurred
at station 8 where attached sea grasses were more
abundant than at any other station. These grasses
probably were effective in trapping detritus as it was
flushed from the marsh by tidal action. Because almost
no detritus was observed in the sediments from the
open bay station, we think that most of the detritus in
the altered area originated in the adjacent marsh.
MILLILITERS
Yy 7 | 7),
_A_ m= lA | ZAE\Z. :
1 4 6 7 8 1
Figure 3.—Average volumes of organic matter (cord grass roots, sub-
merged marine grasses, and organic detritus combined) by station and
zone, March-October 1969.
Seasonally, the volume of plant material changed
little in the shore zone at stations 6, 7, and 8 where
cord grass roots were abundant, but at the stations
where detritus dominated the volume was much great-
er during spring and early summer than in late summer
and fall (Fig. 4).
Dissolved oxygen values were consistently lower at
station | in the deadend canal than at the other stations
(Fig. 5); the observed values remained below 3
m1/liter from June through mid-August. During this
period, zero oxygen values were observed at station 1
on three occasions (Corliss and Trent, 1971).
Loa itr T T T T io Teen T Tea ol aa aaa T ~ |
ah 77»SHORE STATION
ae | ile
O.5-F
OF
1OF
5h
(245})
5.0F
40
20
nw-
MILLILITERS
10
ob wr > —P |
QE 4 1 1 4 1 4 4 = 1 4 1 JL 1 1 |
25 8 226 20 3 17 | 15 29 l2 26 9 23 7 2i
MAR APR. MAY JUN JUL AUG SEP Oct.
Figure 4.—Average volume of organic matter by date, station, and
zone.
LILITERS PER LITER
Figure 5.—Average dissolved oxygen by date for all six stations com-
bined and for the stations having the highest and lowest average val-
ues.
RELATIVE ABUNDANCE
During the study, 8,397 specimens of macroinver-
tebrates belonging to four phyla were collected (Table
1). The numbers of animals caught by station, family
(phylum for nemerteans), date, and zone, and the vol-
ume of the two samples by station and zone are shown
in Appendix Tables 1 and 2. Polychaetes (Annelida)
Table 1.—List of taxonomic groups and total numbers of specimens collected by station and zone.
Station and zone
Taxonomic group 1 4 6 7 8 10 Stations combined
Sie S} Ce S} Ce Ss} G2 Si (ee S3 (CH S} (Cx
Phylum Annelida: 480 292 1,631 234 399 174 281 309 S19 ees (*) 275 3,110 2,465
Class Polychaeta:
Family:
Nereidae 3 0 ul 3 28 20 68 40 36 247 ee) 15 142 102
Terebellidae 0 0 3 1 i 2 3} 6 8 IP ©) 2 21 23
Capitellidae 432) 2905 136015 9 9229 348 148 204 241 268 ~~ 1,100 + 1G) 188 2,853 2,196
Maldanidae 0 0 0 0 0 0 2 0 0 6) 1G): 0 2 6
Arenicolidae 0 0 0 0 7 0 0 2 3 0 (3) 0 10 D
Unidentified 45 D: 20 1 9 4 4 20 4 39 (°) 70 82 136
Phylum Arthropoda: 197 16 11 34 967 123 240 235 322 333 (°) 4 1,737 745
Class Crustacea:
Family:
Ampeliscidae 0 0 9 32 318 98 92 228 64 313 (3) 4 483 675
Corophiidae 186 0 1 D) 257 17 18 5 62 qf (°) 0 524 31
Pinnotheridae 2 0 0 0 0 0 0 0 0 ): 1 0) 0 2 0
Unidentified 9 16 1 0 392 8 130 2 196 13 (°) 0 728 39
Phylum Mollusca: 4 0 33 24 13 2 4 4 9 82 (°) 38 63 150
Class Pelecypoda:
Family:
Tellinidae 0 0 8 18 3 0 0 2 0 Si) 6 11 34
Solecurtidae 3 0 4 0 7 0 4 0 6 64 (3) 0 24 64
Mactridae 1 0 0 0 0 0 0 0 0 Oe ©) 0 1 0
Mytilidae 0 0 0 0 2 0 0 0 0 OWN Fe) 0 2 0
Solenidae 0 0 0 0 0 0 0 0 0 Oi) 8 0 8
Semelidae 0 0 21 6 1 2 0 2 3 10 () 24 25 44
Phylum Nemertinea: 14 0 10 6 19 8 14 14 29 105° 1@) 3 86 4]
Shore.
1
2 Center of waterway.
3 No sampling.
were dominant comprising 66.4% of the number and
44.0% of the volume of organisms caught (Fig. 6).
Crustaceans (Arthropoda) were second in number
(29.6%) but lowest in volume (4.4%). Third in abun-
dance (2.5%), but second in volume (40.8%), were cal-
cified pelecypods (Mollusca). Nemerteans (Nemer-
tinea) were lowest in number (1.5%) and third in vol-
ume (10.7%).
The average number of organisms of each phylum
collected and the results of statistical comparisons of
the data by zone and station are shown in Tables 2 and
3. Abundance values were compared between zones at
each station with a paired t-test and between stations
within each zone with a two-way analysis of variance.
The average of two samples was used as the observa-
tion.
Average total catch (all phyla combined) was higher
in the shore zone than in the center zone, but this
difference was not consistent for all phyla or stations
(Fig. 6, Table 2). In the canals (stations 1 and 4), the
average catches for each phylum were greater along
shore than in the center zone with the exception of
crustaceans at station 4. Only 5 of the 16 differences,
however, were statistically significant. Differences in
average catch between zones in the marsh varied
greatly among stations. At station 6 all phyla were
caught in greater numbers along shore, significantly so
for polychaetes and crustaceans. At station 7, average
catches for each phyla were about the same in each
zone. At station 8 the average numbers of polychaetes
and pelecypods were significantly greater in the
center, whereas nemerteans were significantly more
abundant along shore.
Staustically significant differences in abundance be-
tween stations were found for each taxonomic group in
each zone except nemerteans in the shore zone (Table
3). In the shore zone, average catches were highest at
station 4 for polychaetes and pelecypods, at station 6
for crustaceans, and at station 8 for nemerteans. In the
center zone, average catches were highest at station 8
for all groups except nemerteans, for which average
catch was greatest at station 7.
In general, catches of polychaetes and nemerteans
exhibited only slight seasonal variations; crustaceans
AVERAGE NUMBER
AVERAGE VOLUME (ML)
SHORE =]
CENTER=
POLYCHAETA CRUSTACEA PELECYPODA NEMERTEA
Figure 6.—Average numbers and volumes (stations combined) of or-
ganisms caught per sample by taxonomic group and zone, March-
October 1969.
Table 2.—Comparisons between zones by taxon and station of the
mean numbers of organisms collected (paired-comparison /-test).
Zone
Taxon Station Shore Center d.f. t
--— Mean number ----
Polychaeta 1 15.00 9.12 14 1.97
4 50.97 7.31 15 18.46
6 12.47 5.44 15 3} 117)
q/ 8.78 9.66 15 —0.67
8 9.97 36.91 15 1—6.36
Crustacea 1 6.15 0.50 4 1.05
4 0.34 1.06 5 —1.38
6 30.22 3.84 11 22.53
7 7.50 7.34 10 0.05
8 10.06 10.41 9 —0.06
Pelecypoda 1 0.12 0.00 3 16.13
4 1.03 0.75 10 0.90
6 0.40 0.06 5 1.89
7 0.12 0.12 3 0.00
8 0.28 2.56 14 22.31
Nemertinea 1 0.44 0.00 8 16.42
4 0.31 0.19 8 0.77
6 0.59 0.25 9 1.36
a 0.44 0.44 9 0.00
8 0.91 0.32 12 9D)
1% significance level.
5% significance level.
1
2
Table 3.—Comparisons between stations by taxon and zone of the mean numbers of organisms collected (two-way analysis of variance).
Station F-values
Zone Taxon nn
1 4 6 7 8 10 Block Treatment
wana nnn anne nnn nanan nena nee Mean number -----------------------------
Shore Polychaeta 15.00 50.97 12.47 8.78 9.97 (3) 1.19 197.75
Crustacea 6.15 0.34 30.22 7.50 10.06 (3) 13 225 1 4.62
Pelecypoda 0.12 1.03 0.40 0.12 0.28 (©) 22.02 1 4.75
Nemertinea 0.44 0.31 0.59 0.44 0.91 () 0.61 1.68
Center Polychaeta 9.12 7.31 5.44 9.66 36.91 8.59 1.06 18.11
Crustacea 0.50 1.06 3.84 7.34 10.41 0.12 12.40 2 2.30
Pelecypoda 0.00 0.75 0.06 0.12 2.56 1.19 13.24 1 9.93
Nemertinea 0.00 0.19 0.25 0.44 0.32 0.09 1.08 7) shi
1 1% significance level.
? 5% significance level.
and pelecypods exhibited pronounced seasonal varia-
tion (Fig. 7-10). Catches of crustaceans and
pelecypods were highest during spring and early sum-
mer. The significant F-values for blocks (seasons) for
crustaceans and pelecypods and the nonsignificant
F-values for polychaetes and nemerteans for the
pooled station data (Table 3) substantiate these con-
clusions. The exception to this general pattern was the
decline in abundance, or absence, of all groups at sta-
tion 1 in the center zone during June through Sep-
aes
STATION
|
AVERAGE NUMBER CAUGHT PER SAMPLE
i ——— i 2 Se 1 ES
DSMOMoo mon cOlS: olga lle cOMN2t26 89 25am, ell
MAR. APR. MAY JUN JUL AUG SEP. OcT.
Figure 7.—Average number of polychaetes caught per sample by date,
station, and zone.
CRUSTACEA
STATION
SO > SHORE i
° 4
50 CENTER
100
AVERAGE NUMBER CAUGHT PER SAMPLE
Commo ecmomcO -sSmuilalomeconl2meGal Sikes mire
MAR. APR. MAY JUN JUL AUG. SEP. « OCT.
Figure 8.—Average number of crustaceans caught per sample by date,
station, and zone.
tember. This lowered abundance was probably caused
by low dissolved oxygen during this period (Fig. 5).
Benthic organisms in general were most abundant in
areas with sediments composed of low to intermediate
PELECYPODA
nag Lemay
OnouG
Tre ean epee
STATION |
|
SGuOXG DN0UMNG SVOUG DUW0OMD SuouGd
AVERAGE NUMBER CAUGHT PER SAMPLE
| E 10 4
Fe seas a 4 1 ee eT ht z
25 NeW oc} GeO ssruline lyn ee mMI2e26.9econaie vel
MAR. APR. MAY JUN. JUL AUG. SEP. OCT.
Figure 9.—Average number of pelecypods caught per sample by date,
station, and zone.
NEMERTINEA
T rea. T a T T T T T an aaa T ler eat I |
2r STATION 4
IF | D> 4
OL Zire Mn.
r 4
ae 4
4 2 SHORE 4
Qa Ni
s s
gq 0
wn
Reale CENTER
Ww
Q 2+
=
a5 {0}
oO
=)
ee
oO
oar
w |
{ee}
2°
Sie
Bal g
alr
«xO
Wye
4 2b J
ab 10
1
(0) = il
TE
a 1 Ll 4 — 4 4 4 1
D5 Sooo eOL stale tuiSmeomileweGm Om Comic
MAR APR MAY JUN. JUL. AUG. SEP. OcT.
Figure 10.—Average number of nemerteans caught per sample by
date, station, and zone.
amounts of silt and clay (Tables 2 and 3, Fig. 2).
Abundance was higher for each taxon (except for crus-
taceans at station 4) at stations 1, 4, and 6 in the shore
zone where the percentages of silt and clay were lower
than in the center zone. Abundance of each taxon and
sediment compositions were similar between zones at
station 7.
The compositions of plant material, rather than the
compositions of sediments which were similar, proba-
bly caused the large differences in abundance between
zones at station 8. With the exception of crustaceans,
the abundance of organisms was much greater in the
center than in the shore zone. Plant material in the
center zone was mostly sea grasses and attached
algae, whereas along shore the plant material was pre-
dominantly live cord grass roots.
COMPARISONS BETWEEN CANAL,
MARSH, AND BAY
Based on a comparison of mean values for all groups
combined (Fig. 11), benthic organisms were slightly
more abundant numerically and over twice as abun-
dant volumetrically in the marsh than in the canals;
they were least abundant in the bay. When each group
was considered separately, however, numeric and
volumetric abundance by area varied. Polychaetes
were most abundant numerically in the canals, most
abundant volumetrically in the marsh; they were least
abundant in the bay. Crustaceans were over three
times as abundant in the marsh as in the other two
areas. Pelecypods were numerically most abundant in
the bay; volumetrically, they were most abundant in
the marsh. Nemerteans were most abundant in the
marsh and least abundant in the bay.
DISCUSSION
This and other studies (Reish, 1961; Taylor and
Saloman, 1968) imply that production of benthic or-
ganisms will decrease as a result of the type of altera-
tion of the environment studied here. The magnitude
of the reduction, however, is dependent on many fac-
tors.
The type of vegetative productivity, the segment of
the area that is developed, and the configuration of the
canals are of paramount importance in determining
changes in benthic productivity. In many of the es-
tuarine areas in Florida, vegetative production occurs
primarily on the sand flats. Usually, these flats are the
segments which are developed (extrusion of the
shoreline). In the estuaries along the northern Gulf
coast, including Texas, most of the vegetative produc-
tion occurs in the intertidal zone; this and adjacent
inland areas are usually the areas developed (intrusion
of the shoreline). We think, from an ecological stand-
point, that the types of developments (extruded and
intruded) should be reversed in respect to the types of
estuarine area described above. If reversed, the de-
POLYCHAETA
MEAN NUMBER PER SAMPLE
MEAN NUMBER PER M2
MEAN VOLUME (ML) PER SAMPLE
MEAN VOLUME (ML) PER M2
CANAL MARSH BAY
Figure 11.—Average numbers and volumes of benthic macroinverte-
brates caught per sample by area, taxonomic group, and for the
groups combined, March-October 1969.
crease in benthic productivity might be less in each
type of area. The configuration of the canals deter-
mines the rate and extent of eutrophication. Low dis-
solved oxygen resulting primarily from poor water
circulation has been identified as a major problem in
relation to maintaining biological productivity in de-
velopment canals (Reish, 1961; Taylor and Saloman,
1968; Moore and Trent, 1971; Corliss and Trent, 1971;
Lindall, Hall, and Saloman, 1973).
LITERATURE CITED
BRANDT, K.
1897. Das vordringen mariner Thiere in den Kaiser Wilhelm-
Canal. Zool. Jahrb. Ab. Syst., Geol. Biol. Tiere 9:387-408.
CORLISS, J., and L. TRENT.
1971. Comparison of phytoplankton production between
natural and altered areas in West Bay, Texas. Fish. Bull.,
U.S. 69:829-832.
HOLME, N. A.
1964. Methods of sampling the benthos. Jn F. S. Russell
(editor), Advances in marine biology, Vol. 2, p. 171-260.
Academic Press, N.Y.
LINDALL, W.N.,JR.,J. R. HALL, and C. H. SALOMAN.
1973. Fishes, macroinvertebrates, and hydrological conditions
of upland canals in Tampa Bay, Florida. Fish. Bull., U.S.
71:155-163.
MOORE, D., and L. TRENT.
1971. Setting, growth and mortality of Crassostrea virginica in
a natural marsh and a marsh altered by a housing develop-
ment. Proc. Natl. Shellfish. Assoc. 61:51-58.
REISH, D. J.
1956. An ecological study of lower San Gabriel River, Califor-
nia, with special reference to pollution. Calif. Fish Game
42:51-61.
1957. Effect of poilution on marine life. Ind. Wastes 2:114-118.
1961. A study of benthic fauna in a recently constructed boat
harbor in Southern California. Ecology 42:84-91.
TAYLOR, J. L., and C. H. SALOMAN.
1968. Some effects of hydraulic dredging and coastal develop-
ment in Boca Ciega Bay, Florida. U.S. Fish Wildl. Serv.,
Fish. Bull. 67:213-241.
TRENT, W. L., E. J. PULLEN, and D. MOORE.
1972. Waterfront housing developments: their effect on the
ecology of a Texas estuarine area. Jn Mario Ruivo (editor),
Marine pollution and sea life, p. 411-417. Fishing News
(Books) Ltd., Lond.
Appendix Table 1.—Numbers of animals caught by station, date, taxon, and volume of two samples in shore zone.
Annelida Arthropoda Mollusca Nemertinea
Ba lay Seg g 4a 2 5 o)
o = 3 3s r= = 3 z 2 9 s EU Go By oe
3 2D 3 € § 5 = < 3 & g 5 = 3 $2} jude Identified Totals
Gop rey = ° € o os 5 5 o
rah ic eal Bok Ete ye ONinS Ss ae ame only to. (> So
SiationsaDateez, = OS < GOura S So Aes aS) Ane phylum Number Volume
1 25 Mar. 46 as) Ss 2 2230.6
8 Apr. 34 18 3 1 1 Sif tail
22 Apr. 39 1 40 0.1
6 May 76 76 (0.5
20 May 53 2 55 0.4
6June 2 65 3 70 0.3
17 June 31 1 320.4
lJuly 1 20 1 1 1 24 (18
15 July 17 1 18 0.1
28 July 10 1 3 14 0.2
11 Aug. 5 5S 0.1
25 Aug. 3 31 Qi
22 Sept. 1 1 Dd Oil
6 Oct. 9 37 2 48 0.2
20 Oct es 23 4 rte Yaa eG OPEB 04
Total 3 432 45 186" 29 29 Saal 14 695 6.4
4 25 Mar. 115 1 2 1 119 0.6
8 Apr. 119 2. 3 2 126 0.9
22-Apre)) | 125 2 2 I 1 2 1 135 1.6
GiMay a2 Pi iT 1 4 4 123 0.8
20 May 128 1 2 131 1.6
6 June 97 2 1 100 l57/
17 June 2. 1 73 1.5
1 July 145 5 2 1 153 2.5
15 July 1 42 2 45 0.4
28 July 1 84 2 87 0.6
11 Aug. 3 80 2 2 87 «0.7
25 Aug. 118 1 119 0.5
8 Sept 127 I 2 130 = 0.7
22 Sept. 70 3 73 0.5
6Oct. 1 101 11 2 115 0.9
20Och eee. 67 1 poodle zt Ss) Wa
Total 73) 10601 20 9 1 1 Sane 21 10 1,685 16.2
6 25Mar. 6 27 ce 5) 90 lie al 1733. 0.7
8 Apr. 1 32 l 11 120 118 3 286 1.4
22 Apr. 1 13 1538 175 ili 2 2285 04)
6 May 10 185 57 8 260 0.2
20 May 2 ll 49 1 63 © 0.2
6June 7 2 23 1 1 340.4
17 June 1 13 31 2 AT aanO2
lJuly 1 46 1 6 61 0.8
July 2 33 35.0.3
28July 2 27 3 3 350.4
MeAUIe. ly 62 6 9 0.3
25vAug. 1 26 3 1 3 35 2.1
8 Sept. 1 27 1 1 1 31 0.5
22 Sept. 2 23 22 29 «0.3
6 Oct. 1 39 7 1 1 49 1.1
20 Oct. 3 13 4 a 3 23 0.6
Motall 28. 7) 348 Tic SIS) 257, 392 shi 7 2 1 19 1398 enna
10
Appendix Table 1.—Continued.
Annelida Arthropoda Mollusca Nemertinea
Of vou Reekoee ees og P
qs s See Bee gs ce aCe gee ee
Se Jame gs Bo eo. a5 Sake O56 ge eee eo Se Se Identified Totals
ho Tat ese FES ise! EP Wey dies ge ee Oa See ee Ome ac nly t a
: Opes he Gh [she=stape Behe t=] OO oO 6 only to
Station Date Z & Of > <> > TO) ERE GS) Sho MA re VP ann 0 phylum Number Volume
hie 25.Mar: 1 17 104 2 124 0.2
8 Apr. 3 5 1 11 1 21 0.2
22 Apr. 1 7 1 14 4 2 29 0.2
6 May 2 12 2 44 6 3 69 0.6
2OMayei Ul 9 11-28 1 22 1 2 66 1.6
6 June 13 2 3 1 19 0.3
17 June Pd 92 4 1 1 10 0.3
1 July 5 11 2 18 0.2
15 July 4 11 15 0.2
28 July 9 6 1 16 0.7
11 Aug. 10 20 3 1 1 35 Te5
25 Aug. fi 21 1 29 0.6
8 Sept. 3 15 1 19 0.5
22 Sept. 6 36 2 2 46 0.5
6 Oct. 2 14 2 18 0.4
QOOCtN eas a 22 Eo 5 0.1
Total 6833) 204) 2 4 92 «+18 130 4 14 539s 14.1
8 25 Mar. 1 9 1 9 76 96 0.2
8 Apr. 2. 3 1 33 49 2 2 92 0.3
22 Apr. 2 20 1 10 20 68 2 3 128 0.4
6May 4 15 18 3 1 41 0.3
20 May 18 5 23 0.2
6 June 3 9 1 18 2 33 153
17 June 2 22 7 3 34 0.3
1 July 1 26 1 1 29 0.3
15 July 9 9 0.1
28 July 3 3 6 0.4
11 Aug. I -4- 14 5 4 28 0.2
25 Aug. - 6 18 3 27 0.3
Sisepts. 3) 22> 126 4 35 1e2
a2Sept. 4 1 3 2 1 4 42 4.5
6 Oct. 5 31 1 2 39 0.7
1D) Oi oa a oe 2 puliTey 04
Total 36 8 268 324 64 62 196 6 3 29 679 11.1
11
Appendix Table 2.—Numbers of animals caught by station, date, taxon, and volume of two samples in center of waterway.
Annelida Arthropoda Mollusca Nemertinea
o vo ho) 2 o mo} o
Meg ed ea 32 & en &
S| pie Ge ar aos § 2 5 2 Identified Totals
7) o = Sie ss) ae Ones el 9 5 _ ae
Sse Pe Bes aie i eso only to
Station Date 7A (= Oy ee ae >) Cope OS) HE A A DH phylum Number Volume
1 25 Mar. 37 2 39 0.3
8 Apr. 36 14 50 1.4
22 Apr. 91 91 0.2
6 May 2 72 0.8
20 May 21 21 0.4
6 June 15 15 0.4
17 June 6 6 0.1
1 July 8 8 0.1
6 Oct. 2 2 0.1
20 Oct. Pe! ae = 24 Oth
Total 290 2 16 308 3.9
4 25 Mar. 4 6 2. 6 18 1.1
8 Apr. 16 12 2 30 0.4
22 Apr. 5 18 2 2 27 0.2
6 May 31 2 2 35 0.2
20 May 12 12 0.2
6 June 1 13 2 16 0.2
17 June 118 2 120 0.8
1 July 18 2 20 0.2
15 July 1 1 0.2
8 Sept. 6 6 0.1
6 Oct. 4 4 0.2
20Oct) 2 2 1 4 Sak Te a 9 02
Total 3 12229 l 3242. 18 6 6 298 4.0
6 25 Mar. 2 21 2 28 «6 59 0.8
8 Apr. 11 11 3 25 0.4
22 Apr. 12 34 46 0.2
6 May 24 6 2 32 0.2
20 May 11 2 13 0.1
6 June 4 15 2 3 24 0.8
17 June 4 10 25 39 0.6
1 July 2 2 0.1
15 July 2 2 0.1
28 July 12 2D 2 16 0.6
11 Aug. 7 2 9 0.1
25 Aug. 10 10 0.1
8 Sept. 12 2 14 0.1
22 Sept. 6 2 8 0.1
Oct sits. 2 3 8 0.1
Total 20 2 148 4 98 17 8 2 8 307 4.4
12
Appendix Table 2.—Continued.
Annelida Arthropoda Mollusca Nemertinea
By o ws x Os Gel o
A ep er ee eee gee wens
ro oeae (son ae 2 6 6 gf y 8 Identified Totals
Ce Om ao emSe acc Fe 2 Ss 2 2 ¢€ only to ares =
o a) a Ss 6 < ° & o ° io) o
Station Date Z & O = < D Ol =) FAB D phylum Number Volume
7 25 Mar. 2 12 40 3 2 2 2 63 0.4
8 Apr. 18 2 4 P32 98 1.5
22 Apr. 8 42 2 52 0.4
6 May 22 2 62 4 90 0.6
20 May 34 3 10 2 49 0.6
6 June 4 2 2 8 0.2
17 June 26 2 28 0.2
1July 10 10 20 0.8
15 July 14 14 0.4
28 July 4 4 8 0.2
11 Aug. 12 12 24 0.6
25 Aug. 14 14 0.2
8 Sept. 2 15 2 19 0.2
22 Sept. 5 34 if 46 0.4
GOcty 11 4 2 17 0.4
20 Oct. 10 praca Vegas = Phen mal 2 016)
Total 40 6 241 2 20 2283) 50 2 2 2 14 562 Toll
8 25 Mar. 68 6 4 16 4 98 3.0
8 Apr. aD 30 8 2 14 56 1.2
22 Apr. 2 68 12 IGE gf Dil 2 295 5.2
6 May 48 53 2 2 105 2.8
20 May 10 2 94 8 1 Dy 2) 2 121 4.0
6 June 98 2 4 104 7.0
17 June 7) 2 94 22 2 122 1.0
1 July 6 54 6 4 24 2 2 98 2.6
15 July 86 2 2 2 94 4.0
28 July 70 2 72 0.4
11 Aug. 4 132 6 142 16.4
25 Aug. 2 84 2 88 0.4
8 Sept. 40 2 2 44 0.2
22 Sept. 2 2 2 6 0.2
6 Oct. 74 21 2 2 2 101 0.8
AV OSs, es Set Bante A eae) 2260] 1056:
Total 24 12 1,100 6 39 sie 7 il 8 64 10 10 1,606 49.8
10 25 Mar. 5 2 2 5 14 0.4
8 Apr. 2 1 4 7 0.2
22 Apr. 3 3 1 2 2 11 0.8
6 May 16 2 3 7 28 0.6
20 May 1 17 2 20 0.6
6 June 19 19 0.1
17 June 3 1 13 1 18 0.2
1 July 3 8 6 1 18 0.2
15 July 7 2 9 0.3
28 July 36 1 37 0.3
11 Aug. 5 19 3 1 28 0.2
25 Aug. 13 10 23 0.2
8 Sept. 6 7 1 14 0.2
22 Sept. 4 14 1 19 0.3
6 Oct. 8 28 36 0.2
AW OlAi re slits ae Pion SEE at plist 19 0.3
Total 15 2 188 70 4 6 8924! 3 320 Sil
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648. Weight loss of pond-raised channel catfish (Ictalurus punctatus) during holding in
processing plant vats. By Donald C. Greenland and Robert L. Gill. December 1971, iii + 7
pp., 3 figs., 2 tables. For sale by the Superintendent of Documents, U.S. Government
Printing Office, Washington, D.C. 20402.
49. Distribution of forage of skipjack tuna (Euthynnus pelamis) in the eastern tropical
Pacific. By Maurice Blackburn and Michael Laurs. January 1972, iii + 16 pp., 7 figs., 3
tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
650. Effects of some antioxidants and EDTA on the development of rancidity in Spanish
‘mackerel (Scomberomorus maculatus) during frozen storage. By Robert N. Farragut.
February 1972, iv + 12 pp., 6 figs., 12 tables. For sale by the Superintendent of
Documents, U.S. Government Printing Office, Washington, D.C. 20402.
651. The effect of premortem stress, holding temperatures, and freezing on the
biochemistry and quality of skipjack tuna. By Ladell Crawford. April 1972, iii + 23 pp., 3
figs., 4 tables. For sale by the Superintendent of Documents, U.S. Government Printing
Office, Washington, D.C. 20402.
653. The use of electricity in conjunction with a 12.5-meter (Headrope) Gulf-of-Mexico
shrimp trawl in Lake Michigan. By James E. Ellis. March 1972, iv + 10 pp., 11 figs., 4
tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
654. An electric detector system for recovering internally tagged menhaden, genus
Brevoortia. By R. O. Parker, Jr. February 1972, iii + 7 pp., 3 figs., 1 appendix table. For
sale by the Superintendent of Documents, U.S. Government Printing Office, Washington,
D.C. 20402.
655. Immobilization of fingerling salmon and trout by decompression. By Doyle F.
Sutherland. March 1972, iii + 7 pp., 3 figs., 2 tables. For sale by the Superintendent of
Documents, U.S. Government Printing Office, Washington, D.C. 20402.
656. The calico scallop, Argopecten gibbus. By Donald M. Allen and T-. J. Costello. May
1972, iii + 19 pp., 9 figs., 1 table. For sale by the Superintendent of Documents, U.S.
Government Printing Office, Washington, D.C. 20402.
657. Making fish protein concentrates by enzymatic hydrolysis. A status report on
Tesearch and some processes and products studied by NMFS. By Malcolm B. Hale.
November 1972, v + 32 pp., 15 figs., 17 tables, 1 appendix table. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402. :
658. List of fishes of Alaska and adjacent waters with a guide to some of their literature.
_ By Jay C. Quast and Elizabeth L. Hall. July 1972, iv + 47 pp. For sale by the Superinten-
dent of Documents, U.S. Government Printing Office, Washington, D.C. 20402.
659. The Southeast Fisheries Center bionumeric code. Part I: Fishes. By Harvey R.
- Bullis, Jr., Richard B. Roe, and Judith C. Gatlin. July 1972, x] + 95 pp., 2 figs. For sale by
the Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402.
660. A freshwater fish electro-motivator (FFEM)-its characteristics and operation. By
James E. Ellis and Charles C. Hoopes. November 1972, iii + 11 pp., 9 figs.
661. A review of the literature on the development of skipjack tuna fisheries in the cen-
tral and western Pacific Ocean. By Frank J. Hester and Tamio Otsu. January 1973, iii +
13 pp., 1 fig. For sale by the Superintendent of Documents, U.S. Government Printing Of-
fice, Washington, D.C. 20402.
662. Seasonal distribution of tunas and billfishes in the Atlantic. By John P. Wise and
Charles W. Davis. January 1973, iv + 24 pp., 13 figs., 4 tables. For sale by the Superinten-
dent of Documents, U.S. Government Printing Office, Washington, D.C. 20402.
663. Fish larvae collected from the northeastern Pacific Ocean and Puget Sound during
April and May 1967. By Kenneth D. Waldron. December 1972, iii + 16 pp., 2 figs., 1 table,
4 appendix tables. For sale by the Superintendent of Documents, U.S. Government Print-
ing Office, Washington, D.C. 20402.
664. Tagging and tag-recovery experiments with Atlantic menhaden, Brevoortia tyran-
nus. By Richard L. Kroger and Robert L. Dryfoos. December 1972, iv + 11 pp., 4 figs., 12
tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
665. Larval fish survey of Humbolt Bay, California. By Maxwell B. Eldridge and Charles
F. Bryan. December 1972, iii + 8 pp., 8 figs., 1 table. For sale by the Superintendent of
Documents, U.S. Government Printing Office, Washington, D.C. 20402.
666. Distribution and relative abundance of fishes in Newport River, North Carolina. By
William R. Turner and George N. Johnson. September 1973, iv + 23 pp., 1 fig., 13 tables.
For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
667. An analysis of the commercial lobster (Homarus ariericanus) fishery along the coast
of Maine, August 1966 through December 1970. By James C. Thomas. June 1973, v + 57
pp., 18 figs., 11 tables. Fowsale by the Superintendent of Documents, U.S. Government
Printing Office, Washington, D.C. 20402.
668. An annotated bibliography of the cunner, Tautogolabrus adspersus (Walbaum). By
Fredric M. Serchuk and David W. Frame. May 1973, ii + 43 pp. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402.
669. Subpoint prediction for direct readout meteorological satellites. By L. E. Eber.
August 1973, iii + 7 pp., 2 figs., 1 table. For sale by the Superintendent of Documents,
U.S. Government Printing Office, Washington, D.C. 20402.
670. Unharvested fishes in the U.S. commercial fishery of western Lake Erie in 1969. By
Harry D. Van Meter. July 1973, iii + 11 pp., 6 figs., 6 tables. For sale by the Superinten-
dent of Documents, U.S. Government Printing Office, Washington, D.C. 20402.
671. Coastal upwelling indices, west coast of North America, 1946-71. By Andrew
Bakun. June 1973, iv + 103 pp., 6 figs., 3 tables, 45 appendix figs. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402.
672. Seasonal occurrence of young Gulf menhaden and other fishes in a northwestern
Florida estuary. By Marlin E. Tagatz and E. Peter H. Wilkins. August 1973, iii + 14 pp., 1
fig., 4 tables. For sale by the Superintendent of Documents, U.S. Government Printing Of-
fice, Washington, D.C. 20402.
673. Abundance and distribution of inshore benthic fauna off southwestern Long Island,
N.Y. By Frank W. Steimle, Jr. and Richard B. Stone. December 1973, iii + 50 pp., 2 figs.,
5 appendix tables.
674. Lake Erie bottom trawl explorations, 1962-66. By Edgar W. Bowman. January 1974,
iv + 21 pp., 9 figs., 1 table, 7 appendix tables.
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ctober 1974
NOAA TR NMFS SSRF-681
DIVISION OF FISHES
U.S. NATIONAL MUSEUM
NOAA Technical Report NMFS SSRF-681
U.S. DEPARTMENT OF COMMERCE FEB 5 1973
National Oceanic and Atmospheric Administration
National Marine Fisheries Service
Physiological Response of the
Cunner, Jautogolabrus adspersus,
to Cadmium
NOAA TECHNICAL REPORTS . '
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619. Macrozooplankton and small nekton in the coastal waters off Vancouver Island
(Canada) and Washington, spring and fall of 1963. By Donald S. Day, January 1971, iii +
94 pp., 19 figs., 13 tables.
620. The Trade Wind Zone Oceanography Pilot Study. Part IX: The sea-level wind field
and wind stress values, July 1963 to June 1965. By Gunter R. Seckel. June 1970, iii + 66
pp., 5 figs.
621. Predation by sculpins on fall chinook salmon, Oncorhynchus tshawytscha, fry of
hatchery origin. By Benjamin G. Patten. February 1971, iii + 14 pp., 6 figs., 9 tables.
622. Number and lengths, by season, of fishes caught with an otter trawl near Woods
Hole, Massachusetts, September 1961 to December 1962. By F. E. Lux and F. E. Nichy.
February 1971, iii + 15 pp., 3 figs., 19 tables.
623. Apparent abundance, distribution, and migrations of albacore, Thunnus alalunga,
on the North Pacific longline grounds. By Brian J. Rothschild and Marian Y. Y. Yong.
September 1970, v + 37 pp., 19 figs., 5 tables.
624. Influence of mechanical processing on the quality and yield of bay scallop meats. By
N. B. Webb and F. B. Thomas. April 1971, iii + 11 pp., 9 figs., 3 tables.
625. Distribution of salmon and related oceanographic features in the North Pacific
Ocean, spring 1968. By Robert R. French, Richard G. Bakkala, Masanao Osako, and Jun
Ito. March 1971, iii + 22 pp., 19 figs., 3 tables.
626. Commercial fishery and biology of the freshwater shrimp, Macrobrachium, in the
Lower St. Paul River, Liberia, 1952-53. By George C. Miller. February 1971, iii + 13 pp., 8
figs., 7 tables.
627. Calico scallops of the Southeastern United States, 1959-69. By Robert Cummins, Jr.
dune 1971, iii + 22 pp., 23 figs., 3 tables.
628. Fur Seal Investigations, 1969. By NMFS, Marine Mammal Biological Laboratory.
August 1971, 82 pp., 20 figs., 44 tables, 23 appendix A tables, 10 appendix B tables.
629. Analysis of the operations of seven Hawaiian skipjack tuna fishing vessels, June-
August 1967. By Richard N. Uchida and Ray F. Sumida. March 1971, v + 25 pp., 14 figs.,
21 tables. For sale by the Superintendent of Documents, U.S. Government Printing Of-
fice, Washington, D.C, 20402.
630. Blue crab meat. I. Preservation by freezing. July 1971, iii + 13 pp., 5 figs., 2 tables.
II. Effect of chemical treatments on acceptability. By Jurgen H. Strasser, Jean S. Lennon,
and Frederick J. King. July 1971, iii + 12 pp., 1 fig., 9 tables.
631. Occurrence of thiaminase in some common aquatic animals of the United States
and Canada. By R. A. Greig and R. H. Gnaedinger. July 1971, iii + 7 pp., 2 tables.
632. An annotated bibliography of attempts to rear the larvae of marine fishes in the
laboratory. By Robert C. May. August 1971, iii + 24 pp., 1 appendix I table, 1 appendix 0
table. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
633. Blueing of processed crab meat. II. Identification of some factors involved in the
blue discoloration of canned crab meat Callinectes sapidus. By Melvin E. Waters. May
1971, ili + 7 pp.. 1 fig.. 3 tables.
634. Age composition. weight, length, and sex of herring, Clupea pallasii, used for reduc-
tion in Alaska, 1929-66. By Gerald M. Reid. July 1971, iii + 25 pp., 4 figs., 18 tables.
Continued on inside back cover.
635. A bibliography of the blackfin tuna, Thunnus atlanticus (Lesson)
Beardsley and David C. Simmons. August 1971, 10 pp. For sale by the Su
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636. il pollution on Wake Island from the tanker R. C. Stoner. By
Gooding. May 1971, iii + 12 pp., 8 figs., 2 tables. For sale by the Superinten
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637. Occurrence of larval, juvenile, and mature crabs in the vicinity of Beaufo
North Carolina. By Donnie L. Dudley and Mayo H. Judy. August 1971, iii + 10 pp.,
5 tables. For sale by the Superintendent of Documents, U.S. Government Printi
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638. Length-weight relations of haddock from commercial landings in
1931-55. By Bradford E. Brown and Richard C. Hennemuth. August 1971, v
figs., 6 tables, 10 appendix A tables. For sale by the Superintendent of Doi
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639. A hydrographic survey of the Galveston Bay system, Texas 1963-66. By
W. L. Trent, and G, B, Adams. October 1971, v + 13 pp., 15 figs., 12 tables. Foi
Superintendent of Documents, U.S. Government Printing Office, Washi
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640. Annotated bibliography on the fishifg industry and biology of th
Callinectes sapidus. By Marlin E. Tagatz and Ann Bowman Hall. August 1971,
sale by the Superintendent of Documents, U.S. Government Printing Office,
D.C. 20402.
641. Use of threadfin shad, Dorosoma petenense, as live bait during experi
and-line fishing for skipjack tuna, Katsuwonus pelamis, in Hawaii. By rt
Iversen. August 1971, iii + 10 pp., 3 figs., 7 tables. For sale by the Superinte
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642. Atlantic menhaden Brevoortia tyrannus resource and fishery—analysis 0}
By Kenneth A. Henry. August 1971, v + 32 pp., 40 figs., 5 appendix figs., ta
appendix tables. For sale by the Superintendent of Documents, U.S. Government Prir
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643. Surface winds of the southeastern tropical Atlantic Ocean. By John M. S!
Merton C. Ingham. October 1971, iii + 20 pp., 17 figs. For sale by the Supe!
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644. Inhibition of flesh browning and skin color fading in frozen fillets f yel
snapper (Lutzanus vivanus). By Harold C. Thompson, Jr., and Mary F hi
Government Printing Office, Washington, D.C. 20402.
645. Traveling screen for removal of debris from rivers. By Daniel W. Bat
Murphey, and Martin G. Beam. October 1971, iii + 6 pp., 6 figs., 1 tab 01
Superintendent of Documents, U.S. Government Printing Office, Was!
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646. Dissolved nitrogen concentrations in the Columbia and Snake River
their effect on chinook salmon and steelhead trout. By Wesley J. Ebel. Aug
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647. Revised annotated list of parasites from sea mammals caught off
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ATMOSp
NO i,
oP Ac
UP
OC,
WATIONA,
e
NOAA Technical Report NMFS SSRF-681
Physiological Response of the
Cunner, Tautogolabrus adspersus,
to Cadmium
YOUTOn,
e Ye
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< Zi
2 SEATTLE, WA
we =
October 1974
e7a ~~
/?> g1° For sale by the Superintendent of Documents, U.S. Government Printing Office
6-1 Washington, D.C. 20402
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CONTENTS
Page
Introduction and experimental design, by Anthony Calabrese, Ries
SuColliervandJamesi wMillerie ss. ee eee eens e 1
Uptake of cadmium by organs and tissues, by Richard A. Greig, Al-
bert: EyAdams-sandiBetty7AmNelsonm a) .75 =. sche aaa e ee ee 5
Changes in osmoregulation and oxygen consumption, by Frederick
P. Thurberg and Margaret A.Dawson ................ 11
Effects on the immune system, by Richard A. Robohm and Maureen
HYeIN ACK OWS Kittens fo eit cc nih eee eae ineatemnen mrire, tt Rober ai eaR ees 15
Observations on the biochemistry, by Edith Gould and John J. Karo-
NO i. SERNtet ee ERE ETL. Learn | PRES neers ne ch eae ae Rams Be 21
Histopathology, by Martin W. Newman and Sharon A. MacLean .. . 27
iil
SUMMARY ABSTRACT
The cunner, Tautogolabrus adspersus, was exposed to six concentrations of cadmium, as
cadmium chloride (CdCl, -2'2 H, O), for 96 hr. At the end of this exposure period, tests of blood serum
osmolality and gill tissue oxygen consumption were performed. High levels (48 ppm) of this metal
resulted in abnormally high serum osmolality, and an exposure as low as 3 ppm reduced the normal
rate of oxygen consumption. Both of these parameters may be related to observed tissue damage.
The histopathological effects of acute exposure of the cunner to cadmium were manifested in the
kidney, intestine, hemopoietic tissue, epidermis, and gill. Few significant changes were noted in fish
exposed to concentrations less than 48 ppm. The results implicate renal failure as the probable cause
of death subsequent to acute exposure to cadmium.
Clearance of intracardially injected bacteria from the blood of cunners exposed to 12 ppm cad-
mium was examined. The rate of bacterial uptake in the cells of the liver and spleen was increased,
but the bacterial death rate within these cells was decreased. Exposure of fish at 3 to 24 ppm failed to
influence antibody production against sheep red blood cells.
The activity of two liver enzymes changed significantly with exposure to cadmium. Aspartate
aminotransferase was lower in the exposed fish, and a magnesium-linked oxidoreductase in exposed
fish required 10 times as much added magnesium to reach the same level of activity as in the control
fish.
Chemical analyses were made for uptake and clearance of cadmium from exposed cunners. In the
uptake study, cadmium residues averaged 8.5 times higher in liver than in gills. In the clearance
study, substantial reductions in cadmium residues were found in the gills and blood of fish held in
clean seawater for 6 wk after exposure to cadmium, as compared to fish sacrificed immediately after
exposure. Muscle and carcass samples from the “‘cleared’’ fish showed little reductions in cadmium
levels.
ACKNOWLEDGMENT
The authors of each section of this study thank Rita S. Riccio for her critical reading and typing of
this manuscript.
iv
Physiological Response of the Cunner, Tautogolabrus adspersus,
To Cadmium.
I. Introduction and Experimental Design
ANTHONY CALABRESE, RIES S. COLLIER, and JAMES E. MILLER’
INTRODUCTION
Like most metals, cadmium is stable and does not
degrade in the environment. Thus, as increasing
amounts of cadmium are refined, more and more of it
is circulated in the environment, and increasing
amounts may reach man. Cadmium becomes an air
and water pollutant, through a variety of industrial
processes, and is being used in increasing amounts by
the storage battery, plastics, plating, and petroleum
industries (U.S. Council on Environmental Quality,
1971). There is no evidence that cadmium is
biologically essential or beneficial but, on the other
hand, has caused severe human health problems
(McKee and Wolf, 1971). Since cadmium salts are
most likely to be found in estuarine areas that are im-
portant nursery areas for marine fish and shellfish lar-
vae and juveniles, adult marine fish and shellfish are
a potential source of cadmium in the human diet.
Relatively little is known about the effect of cad-
mium on aquatic animals, particularly those in the
marine environment. Most studies on the effect of
cadmium to aquatic species have been performed
with freshwater forms. More recently, however,
research emphasis has been directed toward the effect
of cadmium salts on various marine organisms
(Shuster and Pringle, 1968, 1969; Gardner and
Yevich, 1969, 1970; Jackim, Hamlin, and Sonis, 1970;
Hisler, 1971; Eisler, Zaroogian, and Hennekey, 1972;
Calabrese et al., 1973). These studies have progressed
from the more classical bioassay tests for simply
determining TL, (that concentration of toxicant
causing 50% mortality) to those of physiological stress
caused by sublethal levels of the pollutant being
tested. Studies conducted at sublethal concentrations
of a contaminant material so as to determine
physiological damage to the organism concerned may
be more important than mortality itself. The gradual
elimination of valued marine species by low concen-
trations of toxicants is no less serious than instan-
_ taneous death of those species. In a sense, it is more
_ serious because it is less likely to be obvious and to be
_ traced to its source in time to permit recovery of the
| 4 Milford Laboratory, Middle Atlantic Coastal Fisheries Center,
National Marine Fisheries Service, NOAA, Milford, CT 06460.
environment. Studies of physiological stress caused
by sublethal levels of a toxicant generally require
chronic or long-term exposure, but some physiological
parameters can be examined in short-term tests.
Parameters of physiological damage that can be ex-
amined by long-term exposure include growth, life
span, reproductive success, adaptation to en-
vironmental stress, feeding and mating behavior,
changes in respiration and osmoregulation,
pathological effects, biochemical anomalies, and
genetic alterations.
It is apparent that increases in human population
and technological development are producing serious
stresses on the marine environment, with a resulting
decrease in its effective use. These events, plus
natural events, are fostering conditions that diminish
the harvest of marine resources. The National Marine
Fisheries Service (NMFS), as part of the National
Oceanic and Atmospheric Administration (NOAA), is
concerned about the threat to marine life and is
providing a national focus for marine research to
generate the basic knowledge and understanding of
marine environmental processes required for effective
management of the marine environment and its
resources.
The New York Bight, which is receiving inter-
national attention because of the large amount of
waste material being dumped into it, borders the most
heavily populated and industrialized complex in the
country. Because the Middle Atlantic Coastal
Fisheries Center of the NMFS is located within this
geographical area, it is important that this Center un-
dertake studies to determine the impact of man upon
the living marine resources of this area. This Center,
comprising laboratories in Sandy Hook, N.J.;
Milford, Conn.; and Oxford, Md., has the facilities
and scientists to undertake studies of this type. The
present study was designed to determine the short-
term (96 hr) physiological response of a local fish,
Tautogolabrus adspersus, commonly known as the
cunner, to cadmium. A multidisciplinary approach
was used to determine the following: 1) uptake of cad-
mium into various tissues and organ systems; 2)
changes in osmoregulation and oxygen consumption
rates; 3) changes in enzymological patterns; 4) im-
mune response to various antigens; and 5) induction
of histopathological abnormalities. Results of these
studies are reported in the following sections of this
technical report.
METHODS AND MATERIALS
Collection and Conditioning
Cunners were collected in modified eel pots, a
cylinder of 2-inch-mesh hardware cloth with a funnel
in one end and a hinged door at the other end. Pots
were baited with cracked hard clams, Mercenaria
mercenaria, and fished in 10-25 feet of water in the
Stratford to New Haven, Conn., area of Long Island
Sound. Fish were transported to the laboratory in 15-
liter polyethylene buckets and placed in tanks of flow-
ing seawater at ambient temperature. Before being
exposed to cadmium, the fish were transferred to
tanks of recirculating artificial seawater (Zaroogian,
Pesch, and Morrison, 1969), adjusted to 25 ppt salini-
ty, and maintained at room temperature for at least 1
wk for acclimation. The fish were fed Purina Trout
Chow? during this time, but were unfed for 2 days
prior to and during the experiment.
Exposure
For exposure to cadmium, the fish were placed in
glass aquaria filled to 60 liters, with artificial seawater
(Instant Ocean), which was aerated throughout the
entire exposure period. Cadmium, as cadmium chlo-
ride (CdCl,*2’2 H,O), was added to test aquaria at
concentrations of 0, 3, 6, 12, 24, and 48 ppm of Cd**.
Stock solution for all tests was made up with reagent
grade cadmium chloride dissolved in water at 50 g
Cd?+ per liter and acidified to a pH of 2.5 to maintain
stability. Proper aliquots of the cadmium stock solu-
tion were added immediately prior to the addition of
the artificial seawater to obtain desired cadmium con-
centrations, and aeration was begun a few minutes
prior to the introduction of test fish. Temperatures
ranged from 21° to 25°C and pH levels remained
between 7.3 and 7.6 during the entire study. The
above five concentrations of cadmium were tested in
duplicate, with two aquaria serving as controls, in
each of a series of seven tests. Four cunners were plac-
ed in each aquarium and were observed daily
throughout the 96-hr exposure period, and dead fish
were removed each day. At the termination of each
test the fish were made available to resident scientists
at the Milford laboratory; in addition, specimen
samples were prepared for histopathological examina-
tion by scientists at the Oxford laboratory, Oxford,
Md. Supplementary tests of the same design were
subsequently performed for those research projects
needing further samples.
> Reference to trade names does not imply endorsement by the
National Marine Fisheries Service, NOAA.
bho
Of a total of 500 fish tested, 126 were sampled for
weight and length, averaging 45.2 g in weight and
146.3 mm in length, with a range in length from 115 to
170 mm.
RESULTS AND DISCUSSION
Although the intent of this study was to determine
the physiological response of cunners exposed to sub-
lethal levels of cadmium, rather than to determine
TL,,, some mortality data were nevertheless obtained.
Concentrations of cadmium (in ppm water) tested
and percent mortality (in parentheses) follow: con-
trols 0 (1.8%), 3 (3.5%), 6 (5.4%), 12 (1.8%), 24
(10.7%), and 48 (26.8%). It is obvious from these data
that cunners can tolerate high concentrations of cad-
mium for at least 96 hr. It was also noted in other
phases of this study, however, that those fish exposed
to 48 ppm of cadmium for 96 hr and then placed in
clean seawater all died within a few days. Hisler
(1971) reported that the tautog, Tauwtoga onitis, a fish
in the same family as the cunner, can also tolerate
high levels of cadmium for a short duration. He also
reported that the sheepshead minnow, Cyprinodon
variegatus, and the mummichog, Fundulus
heteroclitus, had TLso values of 50 and 55 ppm,
respectively, for a 96-hr exposure to cadmium.
Although it appears that marine teleosts are relatively
resistant to cadmium, it will be apparent from the
sections that follow that cadmium does, in fact, cause
physiological stress at sublethal concentrations.
LITERATURE CITED
CALABRESE, A., R.S. COLLIER, D. A. NELSON, and J. R.
MacINNES.
1973. The toxicity of heavy metals to embryos of the
American oyster Crassostrea virginica. Mar. Biol. (Berl.)
18:162-166.
EISLER, R.
1971. Cadmium poisoning in Fundulus heteroclitus (Pisces:
Cyprinodontidae) and other marine organisms. J. Fish.
Res. Board Can. 28:1225-1234.
EISLER, R., G. E. ZAROOGIAN, and R. J. HENNEKEY.
1972. Cadmium uptake by marine organisms. J. Fish. Res.
Board Can. 29:1367-1369.
GARDNER, G.R., and P. P. YEVICH.
1969. Toxicological effects of cadmium on Fundulus hetero-
clitus under various oxygen, pH, salinity and temperature
regimes. Am. Zool. 9:1096.
1970. Histological and hematological responses of an estuarine
teleost tocadmium. J. Fish. Res. Board Can. 27:2185-2196.
JACKIM, E., J. M. HAMLIN, and S. SONIS.
1970. Effects of metal poisoning on five liver enzymes in the
killifish (Fundulus heteroclitus). J. Fish. Res. Board Can.
27:383-390.
McKEE, J. E., and H. W. WOLF.
1971. Water quality criteria. 2nd ed. Calif. State Water
Qual. Control Board, Publ. 3-A, 548 p.
SHUSTER, C.N., JR., and B. H. PRINGLE.
1968. Effects of trace metals on estuarine mollusks. Proc.
1st Mid-Atl. Ind. Waste Conf., Univ. Del. (CE-5):285-304.
1969. Trace metal accumulation by the American eastern
oyster, Crassostrea virginica. Proc. Natl. Shellfish. Assoc.
59:91-103.
U.S. COUNCIL ON ENVIRONMENTAL QUALITY.
1971. Toxic substances. U.S. Gov. Print. Off., Wash., D.C.,
25 p.
ZAROOGIAN, G. E., G. PESCH, and G. MORRISON.
1969. Formulation of an artificial sea water media suitable for
oyster larvae development. Am. Zool. 9:1144.
ir
Physiological Response of the Cunner, Tautogolabrus adspersus,
to Cadmium.
II. Uptake of Cadmium by Organs and Tissues
RICHARD A. GREIG, ALBERT E. ADAMS, and BETTY A. NELSON '
ABSTRACT
Cadmium uptake and clearance data were obtained on cunners, Tautogolabrus adspersus,
exposed to various concentrations of this metal in artificial seawater.
In the uptake study, cunners were exposed to 0, 3, 6, 12, 24, and 48 ppm cadmium in
seawater for 4 days. Cadmium residues averaged 8.2 times higher in livers than in gills. At the
48 ppm cadmium exposure level, the livers averaged 195 ppm, as compared to 33.5 ppm for gills
(wet weight values).
In the clearance study, cunners were exposed to 24 ppm cadmium in seawater for 4 days,
after which time half of the fish were placed in clean flowing seawater for 1 mo and half were
sacrificed immediately to determine initial cadmium residue concentrations. Gill, liver, blood,
muscle, and carcass samples were analyzed. Substantial reductions in cadmium residues were
found in the gills and blood of fish held in clean seawater, as compared to samples from fish
sacrificed immediately after exposure to cadmium. Liver samples produced variable results:
livers of fish held in clean seawater for 1 mo contained 62-155 ppm cadmium for four fish and 5-
11 ppm for three fish, as compared to 30-117 ppm for livers from eight fish sacrificed immediate-
ly after exposure to cadmium. Muscle and carcass samples from the ‘‘cleared”’ fish showed very
little reduction in cadmium levels.
INTRODUCTION
Freshwater and marine organisms have the ability
to concentrate metals far in excess of the levels found
in the waters they inhabit. Mollusks and other
shellfish, in particular, selectively concentrate
chemical materials (including metals) up to many
hundreds of times the levels in their environment
(Pringle et al., 1968).
There are three major sources for uptake of metals
by water-inhabiting organisms:
1. The water column (the metals are dissolved in
the water);
2. Particulate matter (the metals are adsorbed to
particles suspended in the water column); and
3. Food (the metals are incorporated in the material
the organism consumes).
There are relatively few reports in the literature
dealing with the uptake of cadmium by marine
organisms. Of those few marine animals studied for
cadmium uptake, mollusks have received the most
attention and finfish the least (Pringle et al., 1968;
Shuster and Pringle, 1969; Eisler, Zaroogian, and
Hennekey, 1972). Several investigators have studied
5 Milford Laboratory, Middle Atlantic Coastal Fisheries Center,
National Marine Fisheries Service, NOAA, Milford, CT 06460.
nN
the toxic effects of cadmium in both freshwater and
marine finfish, but these workers did not include in-
formation on the uptake of cadmium (Ball, 1967;
Gardner and Yevich, 1969, 1970; Roberts, 1963;
Eisler, 1971).
Our objective was to obtain data on the uptake of
cadmium by liver and gill tissues of cunners exposed
to solutions of CdCl,°2'% H,O in artificial seawater. In
addition, information was obtained on the extent of
clearance of cadmium from various tissues and organs
of cunners that were returned to clean seawater after
exposure to cadmium.
METHODS AND MATERIALS
Fish Holding
Uptake study.—The methods of exposure of the
cunner to various concentrations of cadmium chloride
in artificial seawater for the uptake study are describ-
ed in Part I of this collaborative report.
Clearance study.—An independent study was con-
ducted to determine the extent of clearance of cad-
mium from various tissues of the cunner, after the ex-
posed fish were held in clean seawater. Sixteen fish
were exposed to 24 ppm cadmium (as CdCl.*2'2 H,O)
in artificial seawater for 96 hr, and eight fish were
maintained as controls. The fish weighed from 28-89 g
(X = 62.3 g). After the 96-hr exposure, eight of the ex-
posed fish and four control fish were sacrificed to
determine cadmium residues in various parts of the
fish. The remaining eight cadmium-exposed fish and
four control fish were placed in flowing seawater for 1
mo, after which time they were sacrificed for cad-
mium analysis.
Sampling Procedures
Uptake study.—For the uptake study, data were
collected from three separate experiments, in each of
which five different concentrations of cadmium were
used, plus a control. A single pooled sample was made
of the livers from four to five fish per exposure level.
The same sampling procedure was followed with the
gills, which were rinsed in clean seawater immediately
after dissection to avoid possible adherence of cad-
mium chloride to the gill surfaces.
Clearance study.—Muscle, gills, liver, red blood
cells, serum, and carcass were analyzed for cadmium
residues. The sampling was as follows:
Muscle.—Paired fillets were taken from each fish,
skinned, and ground and combined into a single sam-
ple per fish. The skin was added to the carcass sample
(described below).
Gill and liver.—Samples were taken as described
above for the uptake study, except that the samples
were analyzed individually.
Red blood cells and serum.—Blood samples were
pooled from four fish per treatment, except in the case
of the cadmium-exposed fish held 1 mo in clean run-
ning seawater; only seven fish survived, and their
blood pools represented four and three fish, respec-
tively.
Whole blood was taken from the cunner by heart
puncture, placed in a test tube, and allowed to clot at
room temperature for 45-60 min. The serum was
removed from the clot and centrifuged at 350-500 XG
for 10 min. The clarified serum was frozen-stored until
analysis. For the red blood cells samples, the cellular
residue remaining after centrifugation was combined
with the clot.
Carcass.—The remainder of the fish after removal
of the samples described above was called the carcass,
and included the skin removed from the fillets.
Chemical Analyses
For analysis of cadmium in cunner tissues, samples
were placed in 50-ml glass beakers, dried at 110°C for
18 hr, and heated over a Bunsen burner to char the
tissue. The samples were brought to 400°C in a muffle
furnace, removed after 1 hr at that temperature, and
cooled; a small amount of concentrated HNO, was
added to wet the ash, and the samples were returned
to the muffle furnace at room temperature and
brought to 400°C again. This process was repeated
until only a white residue remained in the beakers,
usually after 3-5 additions of HNO;. The residue was
rinsed from each beaker with 10% HNO, and filtered
through Whatman” No. 2 paper. The filtrate was
brought to a final 10-ml volume and subsequently
analyzed with an atomic absorption spectrometer,
employing a deuterium background corrector (Perkin
Elmer? Model 403).
RESULTS
Uptake Study
Cadmium accumulation was far greater in the liver
than in the gills (Table 1). Cadmium concentrations
averaged 8.2 (range 3-15) times higher in liver than in
gill tissue for all concentrations tested. Although there
was substantial variation in results for the three up-
take experiments, the averaged data show a nearly
linear relation for cadmium concentrations in liver
versus cadmium exposure levels (Fig. 1). Variation in
liver-cadmium concentration was greatest at 24 and
48 ppm exposure levels, and least at 3, 6, and 12 ppm
levels.
Uptake of cadmium into gill tissues was curvilinear
in form, as shown in a plot of cadmium concentrations
in gills versus cadmium exposure levels (Fig. 2). Cad-
mium concentrations in gill tissue for 3 and 6 ppm
* Reference to trade names does not imply endorsement by the
National Marine Fisheries Service, NOAA.
0267
200
180
160
140
120
100 O Experiment 1
& Experiment 2
O Experiment 3
@ Average of 1-3
80
60
40
20
Cadmium Concentration In Livers(ppm,wet wt.)
036 12 24 48
Cadmium Concentration In Seawater (ppm)
Figure 1.—Uptake of cadmium by the livers of cunner held 96 hr
in various concentrations of cadmium (as cadmium chloride) in —
artificial seawater.
Table 1.—Uptake of cadmium by livers and gills of cunners, Tautogolabrus adspersus, exposed for 96 hr at various
concentrations of cadmium, as CdCl,*H,0, in artificial seawater.
Concentrations of cadmium in tissues
Concentration
of cadmium
in artificial
Individual experiments
Individual experiments
seawater
(ppm) Ay. 1 2 3 Av. 1 2 3
o--------- ppm wet weight basis’--------- ----------ppm dry weight basis'----------
LIVER TISSUE
0 1.2 0.95 1.65 0.85 5.5 3.6 6.7 ao
3 16.0 13.5 21.5 13.0 54.5 41.0 75.0 47.5
6 34.5 39.0 36.5 27.5 119.5 125.0 131.0 102.0
12 55.0 54.5 65.0 45.0 198.7 182.5 236.0 177.5
24 110.7 143.0 109.0 80.0 390.0 454.0 386.5 329.5
48 195.0 267.0 160.5 157.0 761.3 928.0 744.0 611.5
GILL TISSUE
0 ile Heil 0.9 3 5.4 5.4 5.0 eS,
3 3.0 4.3 2.3 2.5 16.5 21.5 11.5 16.5
6 3.4 5.1 2.4 Pil 17.5 28.0 12.5 13.0
12 6.3 7.5 5.8 5.6 31.8 38.5 28.0 29.0
24 11.9 16.0 12.0 7.8 66.5 88.5 60.5 44.0
48 33.5 43.0 27.5 30.0 171.3 226.5 135.0 152.5
‘ Liver and gill tissues from four to five fish per exposure level were composited and analyzed in duplicate for each experi-
ment. The values shown for individual experiments are averages of the duplicate analyses.
D Experiment 1
Experiment 2
O Experiment 3
@ Average of 1-3
Cadmium Concentration In Gills(ppm,wet wt.)
036 12 24 48
Cadmium Concentration In Seawater (ppm)
Figure 2.—Uptake of cadmium by the gills of cunner held 96 hr
in various concentrations of cadmium (as cadmium chloride) in
artificial seawater.
cadmium exposure levels were virtually the same, and
moderate increases occurred at 12 and 24 ppm levels.
There was a very sharp increase, however, at the 48
ppm level, which may be related to physiological gill
damage. Thurberg and Dawson (this report, Part III)
found a marked depression in oxygen consumption
rates of gill tissues, as well as a breakdown of os-
moregulation in cunners exposed to 48 ppm cadmium.
Clearance Study
Substantial reductions in cadmium residues were
found in the gills, red blood cells, and serum of
cadmium-exposed fish held in clean running seawater
for 1 mo after exposure. In contrast, muscle and car-
cass samples of these fish showed very little reduction
in cadmium residues, as compared to fish examined
immediately after exposure to cadmium (Table 2).
Liver samples produced variable results (Table 2).
Cadmium concentrations in livers of fish examined
immediately after exposure varied from 30 to 117 ppm
(x = 64.2), a nearly 4-fold difference. Cadmium con-
centrations in livers of fish held 1 mo in clean running
seawater after exposure, varied only 5-11 ppm (x =
10) in three of the fish, but varied from 62 to 155 ppm
(x = 92) in the other four fish.
In spite of the pooling of blood samples (which
reduced the number of possible observations), cad-
mium concentrations in both serum and red blood
cells were as variable as those observed in the in-
dividual liver samples (Table 2). All blood pools from
“cleared”? fish, however, had substantially lower
levels of cadmium than those from fish sacrificed im-
mediately after exposure to cadmium.
Gill tissues showed a greater clearance of cadmium
than did the other tissues examined. Gills of fish
sacrificed immediately after exposure contained 6.2-
10.6 ppm cadmium (x = 8.1), and gills of fish held in
Table 2.—Clearance of cadmium from organs and tissues of cunners, Tautogolabrus adspersus, held in flowing
natural seawater for 1 mo after a 96-hr exposure to 24 ppm cadmium, as CdCl,-2!2 H,O.
Cadmium Concentration
Organ
or Immediately after exposure
tissue Average (range)
Flesh 0.17 (0.11 - 0.22)
Liver 64.2 (30.5 - 11722)
Gills 8.1 (Ge 10.6 )
Red blood cells 6.6 (5:2) ‘and £8:0))
Serum 5.9 (5.9 and 6.0 )
Carcass 4.8 (0:9) “and 1622")
Flesh 0.06 -
Liver 0.7 (0.6 and 0.8 )
Gills 0.4 -
Red blood cells 0.4 -
Serum 0.4 =
Carcass 0.09 (0.08 and _ 0.10)
After 1 mo in clean seawater
Average (range)
TEST FISH
0.12 (0.08 - 0.22)
92.0 (62.0 - 155.0 )!
10.0 ( 5.0 5.) allt) )}
3.5 ( 2.8 - 4.7 )
1.8 (0.8 and 2.8 )
1.5 ( 0.7 - 2.3 )
3.5 ( 2.8 - 4.2 )
CONTROL FISH
0.05 -
1.0
0.3
0.4
0.5
0.12
' The livers of 4 fish had cadmium concentrations in the range of 62-155 ppm, whereas the livers of 3 other fish
had cadmium concentrations in the range of 5-11 ppm.
clean running seawater for 1 mo after exposure con-
tained 2.8-4.6 ppm cadmium.
Concentrations of cadmium found in tissues of con-
trol fish (Table 2) were all less than 1 ppm.
DISCUSSION
In the literature, studies of the uptake of cadmium
and other metals by marine animals deal
predominantly with shellfish. Pringle et al. (1968),
who studied the uptake of five metals by several
species of shellfish, examined only the soft-shell clam,
Mya arenaria, for cadmium uptake. Clams exposed to
0.05 ppm cadmium (nitrate) in flowing seawater for 70
days accumulated 8 ppm (whole-body wet weight),
and clams exposed to 0.1 ppm for 56 days ac-
cumulated 9 ppm. Shuster and Pringle (1969) exposed
Kastern oysters, Crassostrea virginica, to cadmium
(nitrate) in flowing seawater for 20 wk. At a 0.1 ppm
exposure level, oysters accumulated 90-100 ppm cad-
mium (whole-body wet weight) within 13 wk, whereas
at a 0.2 ppm exposure level, the animals accumulated
the same concentrations of cadmium within 8-10 wk.
After only 1 wk at 0.1 and 0.2 ppm exposure levels, the
oysters accumulated 7-24 ppm cadmium, with no ap-
parent difference due to exposure levels.
Eisler et al. (1972) studied cadmium uptake by
Eastern oysters; American lobsters, Homarus
americanus; bay scallops, Aquipecten irradians; and
mummichog, Fundulus heteroclitus. The animals
were held for 21 days in flowing seawater containing
10 ppb cadmium (CdCl, «22 H, O). Accumulation of
cadmium was highest in the oysters, with 1.49 ppm
(whole-body wet weight) in the exposed animals and
0.33 ppm in control animals. The mummichog had
whole-body residues of 0.48 ppm cadmium for ex-
posed fish and 0.33 ppm in control fish.
For comparison with these studies, a combined es-
timate of all data was made for whole-body residues in
cunners exposed to 24 ppm cadmium for 96 hr (see
clearance study, Table 2). The whole-body residues
were calculated to be 2-4 ppm (wet weight) for ex-
posed fish and 0.1-0.2 ppm for control fish. Although a
direct comparison of these data with those cited above
for oysters is not possible, it would appear that cunner
accumulates cadmium to a much lesser extent than
the oyster.
LITERATURE CITED
BALL, I. R.
1967. The toxicity of cadmium to rainbow trout (Salmo
gairdnert Richardson). Water Res. 1:805-806.
EISLER, R.
1971. Cadmium poisoning in Fundulus heteroclitus (Pisces:
Cyprinodontidae) and other marine organisms. J. Fish. Res.
Board Can. 28:1225-1234.
EISLER, R., G. E. ZAROOGIAN, and R. J. HENNEKEY.
1972. Cadmium uptake by marine organisms. J. Fish. Res.
Board Can. 29:1367-1369.
GARDNER, G. R., and P. P. YEVICH.
1969. Toxicological effects of cadmium on Fundulus hetero-
clitus under various oxygen, pH, salinity and temperature
regimes. Am. Zool. 9:1096.
1970. Histological and hematological responses of an estuarine
teleost tocadmium. J. Fish. Res. Board Can. 27:2185-2196.
PRINGLE, B. H., D. E. HISSONG, E. L. KATZ, and
S.T. MULAWKA.
1968. Trace metal accumulation by estuarine mollusks.
J. Sanit. Eng. Div., Proc. Am. Soc. Civ. Eng. 94:455-475.
ROBERTS, H.
1963. Cadmium toxic to rainbow trout. Prog. Fish-Cult.
25:216.
SHUSTER, C. H., and B. H. PRINGLE.
1969. Trace metal accumulation by the American eastern
oyster, Crassostrea virginica. Proc. Natl. Shellfish. Assoc.
59:91-103.
THURBERG, F. P., and M. A. DAWSON.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. II]. Changes in osmoregulation
and oxygen consumption. Jn Physiological response of the
cunner, Tautogolabrus adspersus, to cadmium, p. 11-13.
NOAA Tech. Rep. NMFS SSRF 681.
Physiological Response of the Cunner, Tautogolabrus adspersus,
to Cadmium.
III. Changes in Osmoregulation and Oxygen Consumption
FREDERICK P. THURBERG and MARGARET A. DAWSON!’
ABSTRACT
The cunner, Tautogolabrus adspersus, was exposed to various concentrations of cadmium,
as cadmium chloride (CdCl, -2'’2 H,O), for 96 hr. At the end of this exposure period tests of blood
serum osmolality and gill tissue oxygen consumption were performed. High levels (48 ppm) of
this metal resulted in an abnormally high serum osmolality and an exposure as low as 3 ppm
reduced the normal rate of oxygen consumption. Both of these parameters may be related to
observed tissue damage.
INTRODUCTION
Cadmium, which is neither essential nor beneficial
to aquatic organisms (McKee and Wolf, 1971), has
been detected in increasing amounts in the tissues of a
number of such animals (Mullin and Riley, 1956;
Peden et al., 1973). The use of cadmium in a variety of
industrial processes has increased in recent years,
making this metal an immediate concern as an en-
vironmental pollutant (U.S. Council on Environmen-
tal Quality, 1971; Dean, Bosqui, and Lanouette,
1972). A number of investigators have demonstrated
the toxicity of this metal to aquatic animals (Eisler,
1971; Calabrese et al., 1973; Collier et al., in press);
however, little is known of the sublethal effects of cad-
mium on finfish. Other metals have been shown to
alter serum osmolality and respiration of freshwater
fish (McKim, Christensen, and Hunt, 1970; Lewis
and Lewis, 1971). Few experiments in this area have
been conducted with metals and marine fish and
fewer still with cadmium as the test pollutant. The
present study was undertaken to determine the effect
of cadmium on osmotic regulation and oxygen con-
sumption in the cunner, Tautogolabrus adspersus.
METHODS AND MATERIALS
Cunners were exposed to 0, 3, 6, 12, 24, and 48 ppm
cadmium for 96 hr by the method of Calabrese,
Collier, and Miller (this report, Part I). At the end of
this exposure period, a blood sample was drawn by
heart puncture using a scalpel and a disposable
Pasteur pipette. Pooled blood samples from three to
4 Milford Laboratory, Middle Atlantic Coastal Fisheries Center,
" National Marine Fisheries Service, NOAA, Milford, CT 06460.
four fish per cadmium concentration were collected in
chilled 15-ml centrifuge tubes and spun at 1,720 X g
for 20 min at 4°C. The osmolality in milliosmoles per
Kg H,O (mOsm) of 0.2-ml serum samples was read on
an Advanced 3L Osmometer.’ Gill tissues from these
same fish were dissected out and placed in 15-ml
Warburg-type flasks chilled on ice; each flask con-
tained 5 ml of cadmium-treated seawater from the
tank from which the fish were removed. Oxygen con-
sumption was monitored over a 4-hr period in a Gilson
Differential Respirometer at 20°C. Oxygen consump-
tion. rates were calculated as microliters of oxygen
consumed per hour per milligram dry weight of gill
tissue (yl/hr/mg) corrected to microliters of dry gas at
standard temperature and pressure.
RESULTS
Cunners exposed to 3-24 ppm cadmium for 96 hr
showed no change in serum osmolality from the nor-
mal value of approximately 340 mOsm determined in
control fish. This value is lower than that of the sur-
rounding seawater (630 mOsm). Osmoregulatory dif-
ficulties were noted in fish exposed to 48 ppm. Serum
osmolality in these fish rose to an average value of 390
mOsm. These data are presented in Figure 1; each
point on the curve represents the mean of six pooled
samples, one from each of six exposures at a given
cadmium concentration.
Cadmium reduced the gill tissue oxygen consump-
tion rates at all concentrations tested. A normal rate
of 0.750 « l/hr/mg was reduced to approximately 0.510
ul/hr/mg after exposure to cadmium at concentrations
? Reference to trade names does not imply endorsement by the
National Marine Fisheries Service, NOAA.
Oxygen Consumption(p2/hr/mg)
(@) 3 6 12 24 48
Cadmium Concentration(ppm)
_ 400 T
5 dl
z
5 375
=
2 T
fo) eos ag Ih
E
=
o
YO 325
Figure 1.—Cunner gill tissue oxygen consumption and serum
osmolality after a 96-hr exposure to cadmium. The upper por-
tion of the graph represents gill tissue oxygen consumption rate
(ul/hr/mg) vs. cadmium concentration; the lower portion
represents serum osmolality (mOsm) vs cadmium. Both curves
show the mean value and standard error.
of 3, 6, 12, and 24 ppm cadmium. Oxygen consump-
tion was slightly higher (0.580 ul/hr/mg) after ex-
posure to 48 ppm, the same concentration at which os-
moregulatory stress was observed. These results are
presented in Figure 1; each point on the curve
represents the mean value of gill tissue oxygen con-
sumption of 18 fish.
DISCUSSION
Marine teleosts maintain a normal blood serum
osmolality considerably below that of the surrounding
medium (Krogh, 1965; Parry, 1966). In the present
study cunner serum osmolality rose considerably
above its normal level after a 96-hr exposure to 48
ppm cadmium. Exposures below this level did not
alter serum osmolality. Teleost kidneys excrete salts
and thus maintain a normal osmotic concentration.
Newman and MacLean (this report, Part VI) detected
gross pathology in the kidneys of certain cunners used
in this study. They reported that kidneys of cunners
exposed to 48 ppm cadmium for 96 hr were nearly
nonfunctional, while those exposed to cadmium con-
centrations below 48 ppm appeared normal. Gill and
gut tissues are also involved in osmoregulatory func-
tion (Krogh, 1965; Prosser and Brown, 1961). Other
investigators have reported gill and kidney tissue
damage in marine teleosts after exposure to cadmium
(Gardner and Yevich, 1970; Eisler, 1971), and New-
man and MacLean (this report, Part VI) noted some
gill and gut damage in cadmium-exposed cunners.
Osmoregulatory difficulty at 48 ppm is, therefore, ap-
parently due to kidney failure, although gill and gut
damage may be contributory.
Gill tissue oxygen consumption was depressed after
exposure to 3-48 ppm cadmium. The slight rise
(although still well below the normal level) in oxygen
consumption at 48 ppm was attributed to increased
osmoregulatory stress at that concentration. The
depression of oxygen consumption may have been due
to gill damage. Newman and MacLean (this report,
Part VI) noted gill tissue abnormalities in fish ex-
posed to cadmium, and Ledgerwood and Brown
(1973) reported cadmium-induced aneurysms in the
gill lamellae of threespine sticklebacks, Gasterosteus
aculeatus. Greig, Adams, and Nelson (this report,
Part II) found elevated levels of cadmium present in
gill tissues of all exposed cunners examined.
In summary, the results of this study demonstrated
two physiological effects of cadmium on the cunner.
High levels (48 ppm) of this metal resulted in an ab-
normally high serum osmolality, and an exposure as
low as 3 ppm reduced the normal rate of oxygen con-
sumption. Both of these parameters may be related to
observed tissue damage.
LITERATURE CITED
CALABRESE, A., R. S. COLLIER, and J. E. MILLER.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. I. Introduction and experimental
design. Jn Physiological response of the cunner, Tautogola-
brus adspersus, to cadmium, p. 1-3. NOAA Tech. Rep.
NMFS SSRF 681.
CALABRESE, A., R.S. COLLIER, D. A. NELSON, and J. R
MacINNES.
1973. The toxicity of heavy metals to embroys of the
American oyster Crassostrea virginica. Mar. Biol. (Berl.)
18:162-166.
COLLIER, R.S., J. E. MILLER, M.A. DAWSON, and F. P.
THURBERG.
In press. Physiological response of the mud crab, Eury-
Panopeus depressus, to cadmium. Bull. Environ. Contam.
Toxicol.
DEAN, J. G., F. L. BOSQUI, and K. H. LANOUETTE.
1972. Removing heavy metals from waste water. Environ.
Sci. Technol. 6:518-522.
EISLER, R.
1971. Cadmium poisoning in Fundulus heteroclitus (Pisces:
Cyprinodontidae) and other marine organisms. J. Fish.
Res. Board Can. 28:1225-1234.
GARDNER, G.R., and P. P. YEVICH.
1970. Histological and hematological responses of an estuarine
teleost tocadmium. J. Fish. Res. Board Can. 27:2185-2196.
GREIG, R. A., A. E. ADAMS, and B. A. NELSON.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. II. Uptake of cadmium by organs
and tissues. In Physiological response of the cunner,
Tautogolabrus adspersus, to cadmium, p.5-9. NOAA
Tech. Rep. NMFS SSRF 681.
KROGH, A.
1965. Osmotic regulation in aquatic animals.
IncsaNeYes 242sp:
LEDGERWOOD, R. D., and G. W. BROWN, SR.
1973. Cadmium induced aneurysms in gill lamellae.
Fish. Univ. Wash., Seattle, Contrib. 375.
LEWIS, S. D., and W. M. LEWIS.
1971. The effect of zinc and copper on the osmolality of blood
serum of the channel catfish, Jctalurus punctatus Rafinesque,
and golden shiner, Notemigonus crysoleucas Mitchill.
Trans. Am. Fish. Soc. 100:639-643.
McKEE, J. E., and H. W. WOLF.
1971. Water quality criteria. 2nd ed. Calif. State Water
Qual. Control Board. Publ. 3-A, 548 p.
McKIM, J. M., G. M. CHRISTENSEN, and E. P. HUNT.
1970. Changes in the blood of brook trout (Salvelinus
fontinalis) after short-term and long-term exposure to
copper. J. Fish. Res. Board Can. 27:1883-1889.
Dover Publ.
Res.
13
MULLIN, J. B., and J. P. RILEY.
1956. The occurrence of cadmium in seawater and in marine
organisms and sediments. J. Mar. Res. 15:103-122.
NEWMAN, M. W., and S. A. MacLEAN.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. VI. Histopathology. Jn Physio-
logical response of the cunner, Tautogolabrus adspersus, to
cadmium, p. 27-33. NOAA Tech. Rep. NMFS SSRF 681.
PARRY, G.
1966. Osmotic adaptation in fishes.
41:392-444.
PEDEN, J.D., J.H. CROTHERS, C.E. WATERFALL, and
J. BEASLEY.
1973. Heavy metals in Somerset marine organisms.
Pollut. Bull. 4:7-9.
PROSSER, C. L., and F. A. BROWN.
1961. Comparative animal physiology. W. B. Saunders,
Phila., 688 p.
U.S. COUNCIL ON ENVIRONMENTAL QUALITY.
1971. Toxic substances. U.S. Gov. Print. Off.,
DC: 25 p:
Biol. Rev. (Camb.)
Mar.
Wash.,
Petite” opel bh
Paks rl ibe eho
Physiological Response of the Cunner, Tautogolabrus adspersus,
to Cadmium.
IV. Effects on the Immune System
RICHARD A. ROBOHM and MAUREEN F. NITKOWSKI'!
ABSTRACT
Two elements of the immune system in cunners, Tautogolabrus adspersus, were examined
after 96-hr exposure to cadmium: 1) clearance of intracardially injected bacteria from the
bloodstream and 2) ability to produce antibody against intraperitoneally injected sheep red
blood cells (SRBC). Exposure to 12 ppm cadmium increased the rates of bacterial uptake in
phagocytes of the liver and spleen but significantly decreased the rates of bacterial killing
within these cells. Exposure of fish at 3 to 24 ppm cadmium failed to influence antibody produc-
tion against SRBC. These results indicate that cadmium affects one aspect of cellular immunity
but not humoral immunity in cunners. This effect may increase susceptibility to infection.
INTRODUCTION
There is evidence that cadmium poisoning in
teleosts disrupts respiratory processes (Schweiger,
1957; Mount and Stephan, 1967) and damages
kidneys (Gardner and Yevich, 1970) and gills (Mount
and Stephan, 1967; Gardner and Yevich, 1970). The
exact mechanisms of these effects are unknown,
although some evidence in liver, kidney, and other
tissues of mammals (Simon, Potts, and Gerard, 1947)
and fish (Jackim, Hamlin, and Sonis, 1970) indicates
that enzyme systems are inhibited. If there is inhibi-
tion in rapidly growing or metabolizing cells, then
cells of the immune system in fish may also be
affected. Identification of such effects, in addition to
being useful as indicators of toxicity, may, in part, ex-
plain environmental mortalities of fish. For example,
if a pollutant can inhibit production of antibody by
the lymphocytes or in some way reduce the effec-
tiveness of reticuloendothelial system phagocytes, fish
may become susceptible to infection. A report by Pip-
py and Hare (1969) linking bacterial infection of
salmon with sudden spikes of copper and zinc levels in
a Canadian river is indicative that metal pollution
may indeed lower fish immunity.
The following investigation was undertaken to
determine whether short-term exposure of the cunner
to sublethal cadmium levels would limit antibody
production or reduce clearance of bacteria from the
blood, spleen, and liver.
Reprinted from NOAA Technical Report NMFS SSRF-681
METHODS AND MATERIALS
Fish Holding and Cadmium Exposure
Cunners, Tautogolabrus adspersus, were captured
from the wild, acclimated to artificial seawater, and
exposed to 3 to 48 ppm Cd*+ (as CdCl,°2'2 H,O), as
described by Calabrese, Collier, and Miller (this
report, Part I). Some fish were immunized (as
described below) with sheep red blood cells (SRBC),
removed from Cd?+ treated water, and held an ad-
ditional 10 days at ambient temperature (23 + 2°C).
In two experiments these immunized fish were held in
20-gal fiber glass aquaria containing recirculating
seawater under constant charcoal filtration. The fish
(with dorsal spine, pectoral fin, or pelvic fin clipped to
denote levels of cadmium treatment) were held nine
or less per tank, and the water changed every second
day. In two later experiments with immunized fish,
fins were no longer clipped, and fish were held in run-
ning seawater (20°-23°C) in 500-gal fiber glass tanks
partitioned off with polyethylene mesh.
Some fish were exposed to 12 ppm Cd** treated
water then removed and held for 1 or 2 days in the
running seawater system. These fish were challenged
by intracardial injection of bacteria, as described
below.
Immunization and Collection of Antisera
Sheep red blood cells in Alsever’s solution were
washed in phosphate buffered physiological saline
(PBS), pH 7.2, until the supernatant was free of
hemoglobin and then diluted in PBS to 0.5% suspen-
sion (packed cell volume/volume PBS). Fish were
weighed and injected intraperitoneally with 0.2 m! of
the suspension per 40-g fish (i.e., 2.6 X 10° ml packed
SRBC/g fish weight). SRBC injections were made at 0
hr, at 0 minus 24 hr or at 0 plus 24 hr relative to the
start of the 96-hr cadmium-exposure period. Second
“booster’’ injections of SRBC were given 7 days after
each initial injection. Serum was drawn from each
fish 6 or 7 days after the second SRBC injection.
Blood was removed from the ventral aorta using a
Pasteur pipette equipped with a small, rubber finger
bulb after nicking the artery with a scalpel blade.
Blood was allowed to clot at room temperature and
the clots to retract at 4°C. After removing red cells
from the serum by centrifugation, sera were stored at
—25°C until assayed.
Hemagglutination Assay
The microtiter system of the Cook Engineering Co.,
Alexandria, Va., was used in making dilutions and
setting up hemagglutination assays. Serial twofold
dilutions of fish serum were made in PBS, pH 7.2,
containing normal rabbit serum (NRS) at 1/100 con-
centration (the rabbit serum was preabsorbed with
SRBC). Washed SRBC were suspended to 1% concen-
tration (v/v) in the PBS-NRS diluent and added to
each serum dilution (0.025-ml serum plus 0.025-ml
SRBC suspension). Controls included normal nonim-
mune fish serum, known positive serum from im-
munized fish and PBS alone. After 2-hr incubation at
room temperature and overnight at 4°C, the titers of
each serum were read by examining the degree of
SRBC agglutination based on a 0 to 4+ rating scale.
The last dilution causing a 2+ agglutination of SRBC
was taken as the titer. A 2+ rather than a 1+ end-
point was used because it gave more reproducible
results. Heat inactivation was not done because it
created a gel in the serum. Hemolysis of red cells was
not a problem because it occurred only after 48-hr in-
cubation—a time long after final readings had been
made.
Growth and Injection of Bacteria
Cells of Bacillus sp (biochemical tests consistent
with Bacillus cereus) were grown well into stationary
phase culture (72 hr) in Trypticase soy broth. The
medium in the culture vessel was constantly agitated
by an air-driven magnetic stirrer while the vessel was
held in a water bath at 37°C. After incubation the
culture consisted of about 70% single cells and 30%
cells attached in pairs (with less than 0.1% spores) by
phase-contrast microscopy. Cells were diluted in
physiological saline, containing 0.1% peptone, and
counted by the pour plate method in Trypticase soy
agar. The following day the bacteria were washed 2X
at 3°C with 0.15 M PBS, pH 7.2, and resuspended to
about 5 X 10° cells/0.1 ml in PBS based on the counts
of the previous day.
Fish which had been held for 24 or 48 hr in running
seawater after termination of 96-hr exposure to 0 ppm
or 12 ppm Cd’* were injected intracardially on a
vol/wt basis with the bacterial suspension. For exam-
ple, a 40-g fish received 0.1 ml, a 60-g fish received
0.15 ml, etc. Actual numbers of viable bacteria in-
jected per 40-g fish varied from 5 X 10° to 5 X 108
because of some cell loss and death during washing in
the PBS diluent. Actual numbers were determined by
bacterial counts on the suspension after injecting all
the fish on a particular day.
Measurement of Bacterial Clearance
Intracardially injected fish were placed in 4-gal
polyethylene pails containing seawater at ambient
temperature (23°C). After 30 or 90 min, bacterial
counts were made of the blood, liver, and spleen using
the following procedures: Blood was removed from the
ventral aorta with a premarked, heparinized Natelson
blood collecting pipette; blood was drawn to the mark
(0.125 ml) using mouth suction on the end of a short
rubber tube with mouthpiece (after first wetting the
heparin with a small amount of blood). Blood was
diluted immediately into a tube of physiological
saline containing 0.1% peptone. In order to minimize
the amount of standing blood (containing bacteria) in
the organs, aspiration of blood was continued until
the fish was bled dry. The liver and spleen were
removed, weighed to three places, and each diluted in
an aqueous solution of 0.5% peptone. In many in-
stances, blood from the pericardial space spilled onto
the liver during removal of that organ. When this
happened, the liver was washed with sterile, distilled
water and blotted on sterile paper toweling prior to
weighing. Each liver and spleen was ground in a
sterile, motor driven tissue grinder with teflon pestle
and further diluted serially in 0.1% peptone-saline
diluent. Plate counts of the blood, liver, and spleen
were made in Trypticase soy agar within 10 min of
organ removal. Values were recorded as percent of the
initial bacterial dose present in each organ.
Calculations for total bacteria in the blood stream
were based on a blood volume of 3% of the fish body
weight (this was approximated from values given by
Thorson, 1961).
RESULTS
Antibody Response to SRBC Injections
In order to examine the possibility that cadmium
could affect protein formation or cell division in newly
produced, immunocompetent cells, fish were given
priming doses of SRBC followed by a second antigen
dose 7 days later. Production of antibody was mea-
sured by ability of fish serum to agglutinate washed
SRBC. Table 1 compares the reciprocal hemagglutina-
tion titers of cadmium-treated fish and fish receiving
no cadmium during the 96-hr holding period. Al-
though antigen was injected into fish on the day be-
fore, the day of, or the day after the start of cadmium
Table 1.—Serum hemagglutination titers of fish immunized by intraperitoneal injection of sheep red
blood cells (SRBC) at the time of 96-hr cadmium exposure. Sera were drawn 2 wk later (1 wk after a
second SRBC injection).
Cadmium concentration (ppm)
0 3-6 12-24
No. No. No.
tested Titer? tested Titer tested Titer
Immunized fish 12 338411 95} 29+6 17 30+5
Nonimmunized fish 9 6+1 12 3841 12 6+1
' Titer is shown as the reciprocal of the mean serum dilution+standard error.
treatment, no differences were noted in eventual
amount of antibody produced; therefore, the data
in the table are grouped as though all antigen injec-
tions were given on the same day.
It may be seen from the table that treatment of fish
with low doses (3-6 ppm) or high doses (12-24 ppm) of
Cd?* did not cause any significant differences in an-
tibody response over those produced by fish not ex-
posed to Cd**. The table also shows that fish had a
natural, low-level agglutinin to SRBC which was dis-
tinguishable from immune agglutination by its lower
titer. About half of the fish tested had no natural
SRBC agglutinins. All fish treated with 48 ppm cad-
mium died within the 2-wk holding period even
though held in fresh, Cd*+free water.
Effects of Cadmium on Bacterial Clearance
Experiments were run to determine whether
cadmium exposure would affect another aspect of im-
munity in cunners, namely, clearance of bacteria by
phagocytic cells of the reticuloendothelial system.
The two primary elements of this system, the liver
and spleen, were examined. Bacteria injected into the
bloodstream via intracardial route were counted after
30 and 90 min for their remaining levels in the blood,
for quantities picked up in the liver and spleen, and
(by subtraction) for quantities killed within the 30-
and 90-min time intervals. Five experiments were run
in which a total of 10 or 11 fish were used for each of
the following four variables: 0 ppm Cd’+ at 30 min,
12 ppm Cd’* at 30 min, 0 ppm Cd?+ at 90 min, and
12 ppm Cd**+ at 90 min. Fish caught and used in
the warm summer months had significantly greater
clearance rates than those caught and used in the
autumn; however, the relationships of the 30- and 90-
min effects and the 0 ppm and 12 ppm Cd?**+ effects
were approximately the same. Therefore, to get a
representative mean (one in which the low count ex-
periments would carry as much weight as high count
experiments), each value within an experiment was
multiplied by a factor for that experiment according
to the following example:
mean of all values
Factor, =
mean for experiment #1
This allowed each experiment to be representative in
calculating the mean while preserving all the treat-
ment effect relationships within that experiment.
Figure 1 depicts the effects of Cd** on clearance of
bacteria. Wilcoxon’s signed rank test for paired obser-
vations was used to determine significance of effects
O ppm CADMIUM 12 ppm CADMIUM
Significant
Differences:
z B (.02
treated fish were matched by weight +30% with non-
treated fish within each experiment). Wilcoxon’s two-
sample test for unpaired observations was used to test
significance between 30- and 90-min time intervals.
Figure 1 shows that within the first 30 min, Cd** ex-
posed fish clear bacteria from the blood stream more
rapidly and take up greater numbers of bacteria in the
liver and spleen than fish not exposed to Cd**
Differences in counts in the blood and liver between
Cd?+ treated and nontreated fish are significant.
However, counts in the cadmium-treated fish change
very little during the next 60 min, i.e., only 6.3% ad-
ditional cells are eliminated. By contrast, the counts
in fish not exposed to cadmium continue to decrease
in the blood, liver, and spleen between 30 and 90 min.
Values for the blood and the total remaining cells in
the blood + liver + spleen are significantly different;
28.7% of the initial bacterial load is eliminated (or
killed) within the 60-min time interval. Thus, at 90
min the total remaining bacteria in the non-Cd?*fish
are significantly lower (P.01) than in the Cd** treated
fish. Statistical analyses were done on all possible
relationships between elements of the 0 ppm and 12
ppm Cd**treated fish and between the 30- and 90-min
time intervals. Any probabilities not shown were
found to be nonsignificant.
DISCUSSION
From the experimental data presented in this
paper, one can conclude that short-term exposure of
the cunner to CdCl, at toxic or near toxic levels does
not affect the production of antibody against SRBC.
Although this conclusion is based upon early antibody
production which, undoubtedly, had not reached a
peak, it seems safe to assume that significant
differences, which were to appear, would show up as a
delay or lag in these early responses. Fish could not be
held for long periods of time because of limitations in
holding space. Although the second SRBC injection
(to hasten the rise in antibody titer) was given after
fish had been in Cd?-free water for 3 days, it is certain
that fish still had high Cd?* levels at this point. The
data of Greig, Adams, and Nelson (this report, Part
Il) show continued high levels of Cd?*+in cunners after
4 wk of holding in Cd?+-free water. Others have shown
that the half-life of Cd** after a single exposure does
is in excess of 200 days in rats, mice, dogs, and mon-
keys (Friberg, Piscator, and Norberg, 1971, p. 66).
The low-level agglutination titers observed in about
half the nonimmunized fish (see titers in Table 1) are
not unusual. Natural or nonspecific agglutinins are
common among fish, as well as other animals. This
does not interfere with immunization experiments as
long as these agglutinins are low enough in titer that
they are not confused with the results of specific im-
mune stimulation.
In contrast to the results on antibody production,
Cd** did have significant effects on uptake and
destruction of bacteria by phagocytes in the liver and
spleen. Cadmium at 12 ppm was used in these studies
because it was the highest level at which there was
consistent survival of fish during post-Cd?*+ holding.
Fish exposed at this level exhibited a more rapid in-
itial uptake of bacteria by cells of the liver and spleen,
but a slower bacterial destruction rate than fish not
exposed to Cd** These results are consistent with con-
clusions drawn by Holmes, Page, and Good (1967)
that the metabolic events accompanying phagocytosis
can be separated into two categories: 1) events
associated with particle uptake and 2) events
associated with degranulation within the phagocyte.
In the present study it appears that Cd?* stimulates
the metabolic events responsible for bacterial uptake
but inhibits degranulation or those events responsible
for delivering bactericidal substances to the inter-
nalized bacteria.
The initial, relatively rapid clearance of bacteria
from the blood cannot be entirely credited to
phagocytosis in the liver and spleen since phagocytic
cells in the kidney and gill tissues could also con-
tribute to bacterial uptake and destruction. However,
cells of the liver and spleen probably take up the ma-
jor portion of the injected antigen. This is assumed for
two reasons: 1) in some instances, the liver and spleen
contained as much as 80% of the tota! bacteria initial-
ly injected and 2) studies in other animals indicate
that cells of the liver and spleen are responsible for
removing the majority of intravenously injected par-
ticulate antigens. Benacerraf, et al. (1957) found that
the liver and spleen of rats removed 85 to 98% of in-
jected carbon or saccharated iron oxide. McCloskey
(1972) showed that the liver and spleen of mice retain-
ed higher proportions of injected bacteria than other
organs. However, data in the latter two references
show the liver as the organ of major uptake; whereas
in cunners, the spleen usually contains more bacteria
than the liver. Since the anterior kidney has been
shown to be a site of antibody production in rainbow
trout (Chiller et al., 1969), it is assumed that this
organ may also take up significant numbers of
bacteria; however, for the reasons already given, it is
unlikely that this uptake in the cunner of is the same
magnitude as that of the liver and spleen.
Lack of significant differences between levels of
bacteria in the liver and spleen at 30 and 90 min (as
seen in Fig. 1) does not indicate lack of activity within
their phagocytic cells. As bacteria are destroyed
within loaded phagocytes, additional bacteria can be
taken from the blood stream to reload the phagocytes.
Hence, the bacterial levels in these organs may appear
to be static when, in fact, there is a rapid turnover.
The destruction rate of the bacteria (28.7%/hr in the
normal fish) can be greatly increased when bacteria
from 18-hr growth cultures are used (rather than 72-hr
cultures); in one experiment (unpublished data)
viable bacteria were reduced so quickly that they
dropped below the counting range in 30 min.
Presumably, the older (72-hr) bacteria are in a more
dormant state, which is less susceptible to an-
tibacterial metabolites.
One clue regarding the mechanism of cadmium ac-
tion is given by this work. It appears that within
phagocytic cells cadmium may prevent the delivery of
lysosomal substances to the phagocytic vacuole or in-
hibit the action of these substances on bacteria. On
the other hand, cadmium does not appear to inhibit
events leading to protein formation. This is indicated
because lymphocytic cells exposed to cadmium in
vivo actively proliferate and produce antibody protein
to the same extent as cells not exposed to cadmium.
This work also suggests one way in which cadmium
could reduce fish populations. Since bacteria are more
slowly killed within phagocytes of cadmium-exposed
fish, it follows that certain marginally pathogenic
bacteria may multiply within phagocytes and even-
tually overwhelm the fish with infection. Studies of
the effects of chemical agents in other animals suggest
a common mechanism for phagocytic dysfunction.
Laurenzi et al. (1963) found reduced clearance of
aerosolized bacteria from lungs of mice exposed to
ethanol, cortisone, or cigarette smoke. Green and
Carolin (1967) found that cigarette smoke inhibited
the capacity of rabbit pulmonary macrophages to in-
activate bacteria. Kass and Finland (1953) reviewed a
large body of literature showing that treatment of
animals with cortisone and other adrenocortical hor-
mones increases severity of bacterial, viral, fungal,
protozoan, and helminth infections. Sidransky,
Verney, and Beede (1965) showed that mice treated
with cortisone or cytotoxic cancer therapy drugs
became highly susceptible to pneumonia from
aerosols of Aspergillus flavus spores. Merkow et al.
(1968) then demonstrated that lysosomes in
phagocytes of cortisone-treated mice failed to fuse
with vacuoles containing the spores. Consequently,
the substances in these lysosomes were not delivered
to the vacuole. Jones and Hirsch (1972) have also
demonstrated absence of lysosomal fusion with
phagocytic vacuoles containing living toxoplasma
parasites. These studies indicate a possible universal
mechanism for shutting off microbicidal activities
within phagocytes. If so, it is likely that a number of
environmental pollutants may be found to cause
similar phagocytic dysfunction in fish.
LITERATURE CITED
BENACERRAF, B., G. BIOZZI, B. N. HALPERN, and C. STIFFEL.
1957. Physiology of phagocytosis of particles by the R.E.S.
In B. N. Halpern, B. Benacerraf, and J. F. Delafresnaye
(editors), Physiopathology of the reticulo-endothelial system,
p. 52-77. Blackwell Sci. Publ., Oxford.
CALABRESE, A., R. S. COLLIER, and J. E. MILLER.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. I. Introduction and experimental
design. In Physiological response of the cunner, Tauto-
golabrus adspersus, to cadmium, p. 1-3. NOAA Tech.
Rep. NMFS SSRF 681.
CHILLER, J.M., H.O. HODGINS, V.C. CHAMBERS, and
R. S. WEISER.
1969. Antibody response in rainbow trout (Salmo gairdneri).
J. Immunol. 102:1193-1201.
FRIBERG, L., M. PISCATOR, and G. NORDBERG.
1971. Cadmium in the environment. CRC Press, Cleveland,
166 p.
GARDNER, G. R., and P. P. YEVICH.
1970. Histological and hematological responses of an estuarine
teleost to cadmium. J. Fish. Res. Board Can. 27:2185-
2196.
GREEN, G. M., and D. CAROLIN.
1967. The depressant effect of cigarette smoke on the in vitro
antibacterial activity of alveolar macrophages. N. Engl. J.
Med. 276:421-427.
GREIG, R. A., A. E. ADAMS, and B. A. NELSON.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. I. Uptake of cadmium by organs
and tissues. In Physiological response of the cunner,
Tautogolabrus adspersus, to cadmium, p. 5-9. NOAA Tech.
Rep. NMFS SSRF 681.
HOLMES, B., A. R. PAGE, and R. A. GOOD.
1967. Studies of the metabolic activity of leukocytes from
patients with genetic abnormality of phagocytic function.
J. Clin. Invest. 46:1422-1482.
JACKIM, E., J. M. HAMLIN, and S. SONIS.
1970. Effects of metal poisoning on five liver enzymes in the
killifish (Fundulus heteroclitus). J. Fish. Res. Board
Can. 27:383-390.
JONES, T.C., and J. G. HIRSCH.
1972. The interaction between Toxoplasma gondii and mam-
malian cells. II. The absence of lysosomal fusion with
phagocytic vacuoles containing living parasites. J. Exp.
Med. 136:1173-1194.
KASS, E. H., and M. FINLAND.
1953. Adrenocortical hormones in infection and immunity.
Annu. Rev. Microbiol. 7:361-388.
LAURENZI, G. A., J. J. GUARNERI, R. B. ENDRIGA, and
J. P. CAREY.
1963. Clearance of bacteria by the lower respiratory tract.
Science (Wash., D.C.) 142:1572-1573.
McCLOSKEY, R. V.
1972. Uptake and killing of Mima polymorpha and Herellea
vaginicola by the reticuloendothelial system of neonatally
thymectomized nonwasted mice. Infect. Immun. 6:21-
26.
MERKOW, L., M. PARDO, S. M. EPSTEIN, E. VERNEY, and
H. SIDRANSKY.
1968. Lysosomal stability during phagocytosis of Aspergillus
flavus spores by alveolar macrophages of cortisone-
treated mice. Science (Wash., D.C.) 160:79-81.
MOUNT, D.I., and C. E. STEPHAN.
1967. A method for detecting cadmium poisoning in fish.
J. Wildl. Manage. 31:168-172.
PIPPY, J. H. C., and G. M. HARE.
1969. Relationship of river pollution to bacterial infection
in salmon (Salmo salar) and suckers (Catostomus
commersoni). Trans. Am. Fish. Soc. 98:685-690.
SCHWEIGER, G.
1957. Die toxikologische einwirkung von schwermetallsalzen
auf fische und fischnahrtiere. /Engl.summ.j/ Arch. Fisch-
ereiwiss. 8:54-78.
SIDRANSKY, H., E. VERNEY, and H. BEEDE.
1965. Experimental pulmonary aspergillosis.
79:299-309.
Arch. Pathol.
20
SIMON, F. P., A. M. POTTS, and R. W. GERARD.
1947. Action of cadmium and thiols on tissues and enzymes.
Arch. Biochem. 12:283-291.
THORSON, T. B.
1961. The partitioning of body water in Osteichthyes:
Phylogenetic and ecological implications in aquatic verte-
brates. Biol. Bull. (Woods Hole) 120:238-254.
Physiological Response of the Cunner, Tautogolabrus adspersus,
to Cadmium.
V. Observations on the Biochemistry
EDITH GOULD and JOHN J. KAROLUS'
ABSTRACT
In the liver of cunner, Tautogolabrus adspersus, exposed to 3 ppm and to 24 ppm Cd for 96
hr, aspartate aminotransferase activity was 71% and 59%, respectively, of the activity in livers
of control fish.
In the livers of cunners exposed to 24 ppm Cd, nicotinamide-adenine dinucleotide reductase
activity required 20 mM Mg for activation of the same order that 2 mM Mg produced in control
livers.
Although individual variation precludes generalization here, what may be a metal-
complexing group of proteins in the serum of cadmium-exposed cunner warrants further elec-
trophoretic study.
INTRODUCTION
In the collective effort to determine the effects of
heavy metals on the marine ecology, relatively little
attention has been directed toward possible
biochemical malfunctions in marine animal tissue.
Yet apart from acute physical trauma, such as gross
occlusion of gill tissue, the earliest response of a
marine animal to physiological challenge by sublethal
concentrations of heavy metals is at the molecular
level.
Normal metabolic response to the ingestion or ab-
sorption of heavy metals is their temporary inactiva-
tion by serum proteins, which sequester and transport
the metals to the liver for further processing for
removal from the body. Furst, Flessel, and Kelly
(1972) observed that noncancer-causing metals, such
as zinc and iron, are carried by a- and _ 6-globulins,
whereas those heavy metals definitely known to be
able to cause cancer, such as nickel and cadmium, are
carried by albumins, the major metal-transport pro-
tein of blood. Serum electrophoretic patterns may
conceivably be a means of detecting an abnormal
proportion of metal-protein complexes.
If the normal biochemical mechanisms are unable
wholly to inactivate the heavy metals, toxic effects
follow. The ionic character of the blood serum
becomes seriously deranged (Lewis and Lewis, 1971),
with consequent osmoregulatory distress (Thurberg
and Dawson, this report, Part III). Key ligand af-
finities of some enzymes, particularly those for which
divalent cations act as positive or negative effectors,
can be distorted by a changing ionic environment,
‘Milford Laboratory, Middle Atlantic Coastal Fisheries Center,
National Marine Fisheries Service, NOAA, Milford, CT 06460.
21
with consequent changes in their capacity to react
(Gould, 1969, 1971).
Such effects on biochemical systems are most readi-
ly assayed by measuring changes in the activity of
their constituent enzymes. Because it is not always
clear what systems are involved, one must look either
to enzymes that are known to sequester metals or to
those that require metals for their proper catalytic
functioning. Jackim, Hamlin, and Sonis (1970), for
example, in their acute-static study of heavy-metal
poisoning in the liver of mummichog, Fundulus
heteroclitus, selected three metal-requiring enzymes
(alkaline phosphatase, xanthine oxidase, and
catalase), the metal-sensitive ribonuclease, and acid
phosphatase, an enzyme involved in mineral
metabolism.
In the multidisciplinary study reported here, the
cunner, Tautogolabrus adspersus, was exposed for 96
hr to varying concentrations of cadmium, a soft Lewis
acid with the capacity to bind strongly and irrevers-
ibly to sulfur groups. It may be well to note here that
the nature of cadmium-protein bonding has been
observed to differ with the duration of actual metal
challenge (Nordberg, Piscator, and Lind, 1971): in
mouse liver shortly after a single injection of cad-
mium, the cadmium-protein complex was of high
molecular weight, whereas in livers of mice surviving
for more than 24 hr after injection, the cadmium was
bound to a protein of low molecular weight—probably
the sulfur protein metallothionein, whose synthesis by
the liver is stimulated by prolonged exposure to cad-
mium (Anonymous, 1972). More importantly, in vitro
binding of cadmium by liver homogenates—which
would be rapid—is nonselective, and will inhibit
sulfhydryl-dependent enzymes, whereas a
metallothionein-cadmium complex has no such in-
hibitory effect. In long-term exposure to sublethal
concentrations of heavy metals, therefore, an animal’s
biochemistry may be adapted in ways that will mask
effects observed in acute-static studies.
In the initial seven experiments of this study,
results of which are published throughout this
collaborative report, skeletal muscle was the only
tissue available in sufficient quantity for biochemical
testing. Although the relatively slow metabolism of
the muscle would not be expected to reflect the
biochemical response of rapidly metabolizing liver, for
instance, the muscle sarcoplasm was, nevertheless,
examined for possible changes in malic enzyme (ME)?
and a-glycerophosphate dehydrogenase activities,
which in vertebrates require manganese and
magnesium, respectively, for optimal activity.
Subsequent experiments provided liver and blood,
where early reaction to physiological challenge would
logically be expected. In one experiment, the test con-
centrations of cadmium were 0, 3, and 24 ppm, with
nine fish at each concentration; the livers were tested
for aspartate aminotransferase (AAT) activity, a trans-
aminase that reflects metabolic stress in vertebrates
(e.g., Amador and Wacker, 1965) and invertebrates
(e.g., Hammen, 1969) alike. In another experiment,
only two concentrations of cadmium were used (0 and
24 ppm), with 14 fish at each concentration; the livers
of these fish were tested for changes in ligand response
of NAD reductase activity. The sera were subjected to
electrophoresis and examined for possible changes in
total-protein patterns that might reflect an increasing
metal-protein fraction, for carbonic anhydrase activi-
ty (a zinc-enzyme), and for esterase activity.
METHODS AND MATERIALS
Cunners were exposed to 0, 3, and 24 ppm cadmium
for 96 hr in experiments subsequent to those described
by Calabrese, Collier, and Miller (this report, Part I),
under the same test conditions. The blood was drawn
as described by Thurberg and Dawson (this report,
Part III); the livers were excised, pooled, and placed
in small plastic pouches from which as much air as
possible was excluded before freezing. In several of the
initial experiments that included more concentrations
of cadmium (Calabrese et al., this report, Part I), the
fillets were cut from the cunner frames, skinned, and
packaged and frozen in the same way as the livers.
Treatment of Tissue
Liver.—The freshly excised livers from three fish
were pooled for each sample and were frozen-stored
* Abbreviations used in this report are: ME=malic enzyme,
E.C.1.1.1.40; @GPdH=alpha-glycerophosphate dehydrogenase,
E.C.1.1.1.8: NAD=nicotinamide-adenine dinucleotide, and NADH=
the reduced form; NADR-Mg=magnesium-dependent NAD re-
ductase activity: and ATT = asparate aminotransferase, E.C.2.6.1.1.
until use, for no longer than 1 wk. The pooled livers
were homogenized with a glass pestle in chilled,
double-distilled water, 1:9 (w/v) for the AAT assay
and 1:4 or 1:9 for the NADR-Mg assay. Homogenates
were centrifuged for 45 min at 14,500 g and 4°C, and
the supernates used as the crude enzyme preparations
(E). For AAT assays of the freshly frozen livers, it was
necessary further to dilute the supernates 1:9 with
iced water, for a final E dilution factor of 100.
Skeletal muscle.—Paired fillets, frozen-stored
from 4 to 6 wk, were pounded to a rough paste with an
iced mortar and pestle and centrifuged for 45 min at
14,500 g and 4°C. No suspending medium was used.
The centrifuged tissue fluid (CTF) served as the en-
zyme preparation for ME and a@GPdH assays.
Blood.—The clotted fresh blood was clarified by
centrifugation for 30 min at 1,720 g and 4° C, and the
resulting serum was used for electrophoresis.
Assay Procedures
The water used in preparing all solutions was
doubly glass-distilled; solutions of substrate and
coenzyme were prepared fresh daily; and the assays
were read on a double-beam, ratio-recording spec-
trophotometer, in an optical cuvette with a 1-cm path
length. Change in absorbance at 340 nm from 30 to
90 sec after the beginning of the reaction was taken
as the unit of measurement (A A*° X 10°/min/0.10 ml
EB).
Aspartate aminotransferase.—The procedure
used was essentially that of Bergmeyer and Bernt
(1963), except for the proportions used of reagent
solutions. No malic dehydrogenase was added. H, the
supernate from the liver homogenate, had a dilution
factor of 100.
Protocol:
Buffer-substrate solution: = 2.70 ml
Phosphate buffer (0.1 M, pH 7.6),
and K aspartate (0.25 M)
NADH solution (10 mg/ml H,O) = 0.10 ml
E preparation = 0.10 ml
The solutions were pipetted into an optical cuvette
and allowed to stand for 10 min at room temperature.
Absorbance was read against a reference cuvette (con-
taining medium with no initial NADH); the reference
mixture was adjusted with small increments of
NADH so that the difference in absorbance between
sample and reference was no greater than 0.600. The
reaction was not started until there was no detectable
oxidation of NADH. Substrate (0.10 ml 0.2 M
potassium a-ketoglutarate) was added to start the
reaction.
Magnesium-dependent NAD reductase.
Protocol: Tris buffer,
0.1 M, pH 9.0 = 1.80 ml
NAD, 13 mM,
10 mg/ml H,O = 0.10 ml
MgCl.°6 H,O, 0.06 M
(concn varied)
H,O
E to start reaction
X ml (0, 0.10, 1.00)
(1.00-X) ml
0.10 ml
The rate of reduction of NAD was followed
spectrophotometrically at 340 nm. The control
cuvette contained everything but the enzyme solu-
tion.
Malic enzyme.—The assay for ME, a measure of
the rate of NADP reduction in the presence of malate,
is based on the work of Ochoa et al. (1948), and has
been described elsewhere (Gould, 1965). The buffer
used here was Tris, 0.1 M, pH 8.0.
a-glycerophosphate dehydrogenase.—This
assay, with a discussion of the cation-dilution
technique, is published in detail elsewhere (Gould,
1969).
Electrophoretic Procedures
Electrophoresis.—Electrophoresis of cunner serum
(ca 3 ul/column) was performed at 4°C using a dis-
continuous buffer system, on 7% polyacrylamide gel
columns, pH 9.1, with sample and stacker gels of 3%
polyacrylamide, pH 5.2. The electrode buffer was Tris
(0.005 M)-Glycine (0.038 M), pH 8.3. Running time
was 60 min at 1mA/column followed by 105 min at
3mA/column, using constant current. Both the gel for-
mularies and the electrophoretic procedure are based
on the work of Davis (1964) and have been fully
described elsewhere (Gould and Medler, 1970).
Stains.—For total-protein patterns, the gels were
stained with amido schwartz 10B, 1% in 7% acetic
acid. They were destained by passive diffusion in
several changes of methanol-glacial acetic acid-
distilled water (5:1:5), for a total of about 20 hr.
For visualization of esterase sites, the gels were
stained with a medium containing «a-naphthyl
butyrate (50 mg in 2 ml acetone to dissolve, then 2 ml
H,0), coupled with Fast Garnet GBC in 46 ml
phosphate buffer, 0.1 M, pH 7.0. Polyvinylpyrrolidone
(PVP) (ca 500 mg) was added to the buffer to aid
solubilization of the dye. The substrate was added to
the dye-PVP-buffer solution immediately prior to use,
and the whole filtered through glass wool. Incubation
was in the dark at room temperature for 45 min.
RESULTS AND DISCUSSION
In homogenates of fresh-frozen livers from cunners
exposed to cadmium, AAT activity was significantly
lower than in the controls (Table 1). Livers of fish ex-
posed to 3 ppm Cd had only 71% of the AAT activity
observed in livers of control fish and in fish exposed to
24 ppm Cd, activity dropped to 59% of the control.
Whether this cadmium-induced drop in activity
represents a simple enzyme block, a depression of
microsomal biosynthesis, or a more involved
mechanism of inhibition cannot be speculated from
these few data; the observations here serve only to in-
dicate possibly profitable areas for further work.
Parenthetically, activity of this transaminase in fresh-
frozen livers had roughly 10 times the activity of livers
frozen-stored for longer than 1 wk. Livers frozen for 2-
8 wk (at ca +5°C) not only had much lower AAT ac-
tivity than the fresh livers, but also had widely
variable rates of loss of activity.
Another fresh cunner-cadmium experimental
series, with 14 fish at 0 ppm Cd and 14 at 24 ppm Cad,
provided livers for a study of what appears to be a
soluble, magnesium-linked NAD reductase (NADR-
Mg). Initially, the assay was intended to be for a-
GPdH, but it was discovered that the magnesium
effect was stronger without the aGP substrate: in the
presence of @GP(10 mM), 2 mM Mg produced only a
1.2% and 20 mM only a 1.8% increase in reductase ac-
tivity over activity with no added magnesium;
whereas with no added substrate, 2 mM Mg produced
58% and 20 mM produced 130% increase in reductase
activity over that with no added magnesium (Table
2). The endogenous substrate pool might be expected
to contribute a strong variable to NADR activity
(although not so much in teleosts as in marine in-
Table 1.—Aspartate aminotransferase activity in liver of cunner,
Tautogolabrus adspersus, exposed for 96 hr to varying concen-
trations of cadmium, 25 ppt salinity. Each value is the change in
absorbance at 340 nm for 1 min under assay conditions and
represents the average of 2 tests. Each sample is a pool of livers
from three fish; the enzyme preparation (E) has a 100X dilution
factor.
AAT activity
Test concen (A A*°X10°/min/
Cd (ppm) 0.10 ml E)
0 ppm Cd 151.5
170.0
152.5
3 ppm Cd 110.0
115.0
24 ppm Cd 84.0
93.5
101.5
Table 2.—Effect of added Mg**on NAD reductase activity with and
without added @-glycerophosphate. E dilution factor is 10x. Sam-
ple is pool of three control livers of cunners, Tautogolabrus adsper-
sus.
NAD reductase activity
(A A*°X10%/min/0.10 ml E)
Final concen without added with @-GP
Mg**(mM) substrate (10 mM final conc)
0 5 41
2 29 48
20 65 75
vertebrates), but the assay protocol here was designed
to reduce that variable as much as possible, by using
the enzyme activity without added magnesium as a
base from which to measure magnesium activation.
Table 3 lists data showing a mild cadmium effect
upon the magnesium activation of cunner liver
NADR: in fish exposed for 96 hr to 24 ppm Cd, an ap-
proximately 10-fold increase in magnesium concen-
tration (20 mM) is required to activate NADR as
much as 2 mM Mg does in the control fish.
In frozen-stored cunner skeletal muscle, what
appeared to be a significant difference in ME proper-
ties of pooled fish exposed to cadmium, as contrasted
with pooled controls, must be ascribed to individual
variation. @GPdH activity was very high, but there
was no apparent difference in properties between the
cadmium-exposed and the control fish.
Electrophoretic studies are as yet inconclusive.
Despite considerable variation in the total-protein
pattern, there is an area of tentatively labeled metal-
protein complexes that appeared in most cases (8
series out of 10) to increase in fish that had been ex-
posed to cadmium (Fig. 1); many more data would be
necessary to establish statistical validity, however,
and individual variation renders this approach
questionable in an acute-static study. Detoxification
mechanisms of this nature may be more prominent in
a chronic study.
Of the serum enzymes tested for isoenzyme varia-
tion, only a-naphthyl butyrate esterase activity
produced a difference in pattern between cadmium-
exposed and control fish; but here again, individual
variation was very strong.
On the whole, the observations made in the course
of this preliminary experimentation seem to point to
further work with metal-activated enzymes and with
the stress-indicator transaminases. The most clear-
cut effects were obtained by using assays designed to
measure the degree of ligand activation or inhibition.
LITERATURE CITED
AMADOR, E., and W. E. C. WACKER.
1965. Enzymatic methods used for diagnosis. In D. Glick
(editor), Methods of biochemical analysis, Vol. 13, p. 265-356.
Interscience Publ., N.Y.
ANONYMOUS.
1972. ‘“‘Itai-itai byd’”’ and other views on cadmium,. Food
Cosmet. Toxicol. 10:249-255.
BERGMEYER, H.-U., and E. BERNT.
1963. Glutamate-oxaloacetate transaminase. In H.-U. Berg-
meyer (editor), Methods of enzymatic analysis, p. 837-845.
Academic Press, N.Y.
CALABRESE, A., R. S. COLLIER, and J. E. MILLER.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. I. Introduction and experimental
Table 3.—Mg-dependent NAD reductase activity in liver of cunner, Tautogolabrus adspersus,
exposed for 96 hr to 24 ppm Cd, 25 ppt salinity. Each value is the average of two tests, and each sam-
ple is a pool of three livers.
Increase in activity (A A**°X10*/min)
effected by addition of Mg** to assay medium
0 ppm Cd
24 ppm Cd
final concen Mg**+(mM)
Homogenate
final concen MG*+(mM)
dilution
factor 0 (Init. A A) 2 20 0 (Init. A A) 2 20
10x 0 (5) 24 60 0 (13) 1 35
10X 0 (9) 29 54 0 (22) 0 26
5X 0 (21) 10 83 0 (23) 4 32
5x 0 (22) 35 56 0 (18) 12 27
2X 0 (23) 26 57 - -
5x ) (18) 20 88 =
5X 0 (20) 24 dl - = =
5x 0 (2) 10 45
12
48
24
Figure 1.—Protein pherogram of serum from cunners, Tautogolabrus adspersus, exposed for 96 hr to 0, 6, 12, 24, and 48 ppm cadmium
chloride, at 25 ppt salinity. Arrow points to area tentatively considered to comprise metal-protein complexes.
design. Jn Physiological response of the cunner, Tautogo-
labrus adspersus, to cadmium, p.1-3. NOAA Tech. Rep.
NMFS SSRF 681.
DAVIS, B. J. ;
1964. Disc electrophoresis — II. Method and application to
human serum proteins. Ann. N.Y. Acad. Sci. 121:404-
427.
FURST, A., C. P. FLESSEL, and M. E. KELLY.
1972. Cited in J. Arehart-Treichel, Sounding out metal
toxicity. Sci. News 102:223.
GOULD, E.
1965. Observations on the behavior of some endogenous
enzyme systems in frozen-stored fish flesh. Jn R. Kreuzer
(editor), The technology of fish utilization, p. 126-128.
Fishing News (Books), Lond.
1969. Alpha-glycerophosphate dehydrogenase as an index of
iced-storage age of fresh, gutted haddock (Melanogrammus
aeglefinus). J. Fish. Res. Board Can. 26:3175-3181.
1971. An objective test for determining whether “‘fresh”’ fish
have been frozen and thawed. Jn R. Kreuzer (editor), Fish
inspection and quality control, p. 72-75. Fishing News
(Books), Lond.
GOULD, E., and M. J. MEDLER.
1970. Test to determine whether shucked oysters have been
frozen and thawed. J. Assoc. Off. Anal. Chem. 53:1237-
1241.
HAMMEN, C.S.
1969. Metabolism of the oyster, Crassostrea virginica.
Zool. 9:309-318.
JACKIM, E., J. M. HAMLIN, and S. SONIS.
1970. Effects of metal poisoning on five liver enzymes in the
killifish (Fundulus heteroclitus). J. Fish. Res. Board Can.
27:383-390.
LEWIS, S. D., and W. M. LEWIS.
1971. The effect of zinc and copper on the osmolality of the
blood serum of the channel catfish, /ctalurus punctatus
Rafinesque, and golden shiner, Notemigonus crysoleucas
Mitchill. Trans. Am. Fish. Soc. 100:639-643.
NORDBERG, G. F., M. PISCATOR, and B. LIND.
1971. Distribution of cadmium among protein fractions of
mouse liver. Acta Pharmacol. Toxicol. 29:456-470.
OCHOA, S., A. H. MEHLER, and A. KORNBERG.
1948. Biosynthesis of dicarboxylic acids by carbon dioxide
fixation. I. Isolation and properties of an enzyme from
pigeon liver catalyzing the reversible oxidative decarboxly-
ation of l-malic acid. J. Biol. Chem. 174:979-1000.
THURBERG, F. P., and M. A. DAWSON.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. III. Changes in osmoregulation and
oxygen consumption. Jn Physiological response of the
cunner, Tautogolabrus adspersus, to cadmium, p. 11-13.
NOAA Tech. Rep. NMFS SSRF 681.
Am.
nt 2
ual
. oh
wth?
Physiological Response of the Cunner, Tautogolabrus adspersus,
to Cadmium.
VI. Histopathology
MARTIN W. NEWMAN and SHARON A. MacLEAN'
ABSTRACT
The histopathological effects of acute exposure of cunner, Tautogolabrus adspersus, to
water containing cadmium chloride were manifested in the kidney, intestine, hemopoietic
tissue, epidermis, and gill. Few significant changes were noted in fish exposed to concen-
trations less than 48 ppm for 96 hr. The results implicate renal failure as the probable cause of
death after acute exposure to cadmium.
INTRODUCTION
The histopathological aspects of this study were
undertaken in the hope of contributing to our
knowledge of the effects of heavy metals at the level of
the cells and tissues. Elucidation of the mechanisms
of observed physiological and behavioral responses,
and development of baseline information useful for in-
terpretation of specimens which may be collected
from naturally occurring mortalities were further
goals. Only Gardner and Yevich (1970) have
systematically examined blood and tissues of a teleost
exposed to cadmium. While the exposure levels used
in that study and the present one were similar, the
fish species and the length of exposure (96 vs. 48 hr)
were different.
MATERIALS AND METHODS
Techniques used in handling the cunner,
Tautogolabrus adspersus, in this study have been
described by Calabrese, Collier, and Miller (this
report, Part I). Blood smears were prepared from
heart blood (see Thurberg and Dawson, this report,
Part III) and stained by Giemsa or Wright’s methods.
Differential white cell counts were performed using
the first 250 leucocytes encountered on each smear.
All thrombocytes seen while counting leucocytes were
also enumerated. Tissue samples were removed for
physiological and biochemical studies (Thurberg and
Dawson; Gould and Karolus, this report, Parts II and
V respectively). The remainder of each fish was fixed
in Davidson’s (AFA) fixative for 48-96 hr then
transferred to 70% EtOH. Tissues were embedded in
paraffin, sectioned at 6 um, and stained with a variety
of techniques including Giemsa, PAS, PAS-Alcian
Blue, Mallory triple-stain, and Perl’s Prussian blue
' Oxford Laboratory. Middle Atlantic Coastal Fisheries Center,
National Marine Fisheries Service, NOAA, Oxford, MD 21654.
27
reaction. Tissues from each of six fish exposed to 0, 6,
12, 24, and 48 ppm cadmium, or a total of 30 fish were
examined.
RESULTS
Intestine
Pathological changes were seen in the intestinal
epithelium of cunners exposed to high concentrations
of cadmium. In the 24 ppm exposure group, there was
some swelling of the intestinal epithelium. At 48 ppm,
five of six fish exhibited varying degrees of
pathological change. The columnar cells were swollen.
Nuclei were hypertrophied and occupied a position
farther from the basement membrane than those of
unexposed fish. Nucleoli became very prominent.
Numbers of mucus secreting cells appeared about
equal to or slightly less than in the control animals. In
two of the above five fish, the intestinal epithelium
was sloughed from the basement membrane in many
places and the lumen contained much cellular debris
and mucus (Fig. 1).
Kidney
One of six fish exposed at the 24 ppm level had
some cloudy swelling in a few scattered areas of the
proximal tubules. At 48 ppm, the kidneys of five of six
fish examined showed some degree of pathological
change. Three of the fish exhibited diffuse tubular
necrosis (Fig. 2), one exhibited focal tubular necrosis,
and one only a few scattered necrotic lesions. The
proximal segments of the tubules appeared to be most
affected. The lumina of more distal areas of the
tubules often contained sloughed epithelial cells or
were filled with a hyaline eosinophilic material (Fig.
3).
Glomeruli appeared normal. The blood spaces in
kidneys of fish exposed to 24 and 48 ppm cadmium
Figure 1.—Intestinal epithelium of a control fish (a) and a fish exposed to 48 ppm cadmium for 96 hr (b). Necrosis and sloughing of
the intestinal epithelium is evident in the experimental animal. Note also that much cellular debris occupies the lumen. 120X;
Mallory’s triple stain.
contained large numbers of cells thought to be im-
mature thrombocytes (Fig. 4). The occurrence of
erythrophagocytosis and of hemosiderin, common in
the kidneys of control fish, was reduced or absent in
the kidneys of the 48 ppm group (Fig. 5).
Gills
The appearance of the gills of cadmium-exposed
fish was quite variable. The following defects were
noted in decreasing order of prevalence: epithelial
hypertrophy, hyperplasia of interlamellar epithelium,
and desquamation. These changes were noted at all
levels of exposure. The gill tissue of some heavily ex-
posed fish appeared normal and some lesions were
seen in the control animals. Some of these lesions may
represent postmortem changes. In view of the exten-
sive variability of gill lesions which could not be cor-
related with exposure levels, little emphasis was placed
on the appearance of this tissue.
Epidermis
The epidermis appeared normal in fish exposed at
levels up to 24 ppm. At 48 ppm swelling of the
epithelial cells and a paucity of mucus secretion was
noted (Fig. 6).
Blood
Obvious qualitative differences noted between
smears from cunner exposed to 24 and 48 ppm cad-
mium and control fish consisted of poikilocytosis and
karoklasis (Figs. 7 and 8).
28
Differential leucocyte counts revealed throm-
bocytopenia and lymphocytopenia. However, the
percentage of neutrophils increased (Table 1).
DISCUSSION
The histological and hematological response of an
estuarine teleost to cadmium has been studied by
Gardner and Yevich (1970) using mummichog, Fun-
dulus heteroclitus. Their experiments involved acute
exposure to 50 ppm cadmium for up to 48 hr. The
findings of the present study are very like those of
Gardner and Yevich. Discrepancies which did arise
might be related to difference in the fish species used,
or length of exposure.
Changes in the intestine of cunners were similar to
those reported in mummichogs. Gardner and Yevich
(1970) found an increase in mucus cell activity while
the present study indicates a normal or slightly
depressed level of activity. This difference may be at-
tributable to the increased exposure time of cunner.
Lymphocytic infiltration of the submucosa was not
observed in the present study.
Kidney
Morphologic changes in the kidney of teleosts
exposed to cadmium have also been documented by
Gardner and Yevich (1970). Pathological changes in
the kidneys of mammals exposed to cadmium are well
known. Foster and Cameron (1963) produced renal
lesions in rabbits with two subcutaneous injections of
CdCl, (9 mg Cd*/kg). These lesions were limited to
Figures 2 and 3.—Kidney of control fish (a) and of fish exposed to 48 ppm cadmium for 96 hr (b). Note diffuse tubular necrosis in 2b.
Tubular epithelium in various stages of degeneration can be seen in 3b. Note the swollen nuclei of still intact epithelial cells and
cellular debris in the dilated lumina. 120 (2), 300 (3); Azure-eosin stain.
Table 1.—Effects of 96-hr exposure to cadmium chloride on differential leucocyte counts of cunners, Tautogolabrus adspersus
(figures are averages, standard deviation in parenthesis).
Leucocytes (%)
Cadmium
exposure No. Mature Small Medium
(ppm) sampled thrombocytes Neutrophils lymphocytes lymphocytes Eosinophils Blasts Monocytes
0 8 69.3 (410.8) 15.6 (+9.1) 11.2 (+3.9) 2.2 (+0.7) 0.7 (+0.7) 1.1 (+1.2) 0
3 9 65.3 (£10.1) 18.5 (+10.3) 10.3 (+4.9) 2.4 (41.4) 0.6 (40.4) 2.9 (438.3) 0
6 8 60.5 (£16.3) 23.7 (+19.1) 9.3 (+5.6) i 3 (42.3) 1.2 (0.5) 2.3 (1.7) <0.1
12 9 65.2 (416.5) 24.1 (+16.3) 7.5 (£4.9) 3 (41.0) 0.6 (+0.5) 1.6 (40.5) <0.1
24 9 56.3 (420.8) 32.1 (419.3) 6.4 (+4.6) i 6 (+0. 9) 0.6 (40.6) 3.0 (+2.6) <0.1
48 10 35.9 (412.4) 50.2 (412.4) 4.5 (44.8) 1.5 (+1.1) 1.4 (41.1) 6.1 (+4.8) 0
29
Figure 4.—Kidney of control fish (a) and of fish exposed to 24 ppm cadmium for 96hr(b). Note difference in the cell populations
occupying the vascular spaces. 480; Azure-eosin stain.
. a” Lee ; owes BEA SS
- ‘ ee ee SS
‘ ‘ ~- Fat + ees
we eu © ey - ve :
g. . : Jf x = a ;
P é nS >
ai ae * Race So ; >
5 < . i tiled ~y
Re : WP is
oid *
> a4 Yr
[ =. < we \ “ty
mek ‘\ * “Ky * x . ws Rite .
wr. Soe 3 . ; -
LJ 2 oe \
> ‘ * \ > ¥ & ss
a * s i + ,
. bs - ‘ = é ‘
. ~ \ ‘ <
. - ~
. ‘ . a * 2 ? .
s ras . a |
7 Prd t ¥ ’ ‘
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5 -- * ¥ eq
Ca Sry > we Wes
w+ ‘ .
Loman |
. nN wt os . i
’ \ * * ;
\ - a abe
< ole
Figure 5.—Kidney of control fish (a) and of fish exposed to 48 ppm cadmium for 96 hr. An abundance of hemosiderin can be seen
in the control fish. 120; Perl’s stain.
the proximal tubules. Similar experimental results centrations in renal cortex reach approximately 200
were obtained by Dalhamn and Friberg (1957); ppm wet-weight, functional and morphological
Ahlmark et al. (1961) found decreased glomerular change occurs in the kidneys of rabbits, rats, and men
filtration rates in human workers exposed to cad- (Friberg, Piscator, and Nordberg, 1971). After ad-
mium. Proteinuria is another common finding in ditional exposure, the functional changes result in the ©
human cadmium workers (Friberg, 1950; Piscator, failure of the kidney to reabsorb the metallothionein, —
1962). Proteinuria occurs subsequent to tubular and increased excretion of cadmium and a lowering of
dysfunction because of decreased protein reabsorption the concentration of the metal in the kidney occurs.
in the proximal tubule. The effect of cadmium poisoning on the amount of
During chronic exposure or early in acute exposure, hemosiderin in the teleost kidney has not been
cadmium is concentrated in the renal cortex in the previously described. Unlike most homeotherms,
form of metallothionein, a cadmium- or zinc- many teleosts normally have a great deal of pigment
containing protein with a molecular weight of about in the kidney, composed of both lipofuscin and
10,000 (Kagi and Vallee, 1960). When cadmium con- hemosiderin. The observation of reduced pigment in
30
Figure 6.—Epidermis of control fish (a) and of fish exposed to 48 ppm (b). Note nuclear hypertrophy and paucity of mucus secreting
cells in 6b. The separation of the epidermis from the deeper layers is probably sectioning artifact. 120; PAS-hematoxylin stain.
the cunners exposed to 48 ppm cadmium may be
caused by the metal’s effect on the hemopoietic
system. Friberg (1950) found no effects in human bone
marrow after respiratory exposure. Wilson, DeKds,
and Cox (1941) found that rats fed a diet containing as
low as 31 ppm cadmium would be anemic. By ad-
ministering iron to rats receiving cadmium in their
diet, anemias were reduced (Friberg, 1955). This in-
dicated that cadmium was not directly blocking
hemoglobin synthesis, but might be interfering with
the uptake of iron by the intestine. If a reduction of
iron absorption was occurring in the cunner, this,
along with the observed decrease in erythrophagocytic
activity, might account for the observed reduction in
hemosiderin in exposed animals.
Gill
The lack of correlation between exposure level and
observed pathologic changes in the gill tissue can be
explained by Gardner and Yevich’s observations on
mummichogs. They noted that gill lesions were focal
in that all gill filaments were not involved, and the
lesions were of a random nature, not limited to
specific areas of the branchial arches.
The importance of the effects of heavy metals on
the respiratory epithelium of fishes is not clear. Skid-
more (1964) believed that death of rainbow trout,
Salmo gairdneri, exposed to acutely toxic solution of
zinc sulphate was caused by tissue hypoxia, resulting
from the damaging effect of the metal on gill
epithelium. Subsequent experimental studies seem to
confirm this hypothesis (Skidmore, 1970). Burton,
_ Morgan, and Cairns (1972) studied levels of lactic and
_ pyruvic acid in muscle and liver of rainbow trout ex-
. posed to acute zinc toxicity and reached conclusions
! similar to Skidmore. Schweiger, cited in Eisler (1971),
concluded that the toxic action of cadmium on several
3]
freshwater fish and invertebrates was due to suffoca-
tion. Friberg et al. (1971) cite examples of respiratory
exposure to cadmium producing lung damage in
mammals at levels which were insufficient to cause
kidney damage. Doudoroff and Katz (1953) are cited
by Mount and Stephan (1967) as listing nine
references attributing the death of fish in solutions
containing heavy metal salts to coagulation of mucus
on the gill or damage to gill tissue. Bilinski and Jonas
(1973) found that exposure of rainbow trout to 11.2
ppm Cd’** for 72 hr resulted in a 66% mortality, but
there was no significant change in the ability of the
gill tissue to oxidise lactate.
Evidence from the present study implicates renal
failure as a cause of death in cunner after acute ex-
posure to cadmium. Although pathological changes
were observed in gill tissue and gills exposed to even
low levels of cadmium seemed to sustain some
physiological damage (Thurberg and Dawson, this
report, Part III), these changes do not seem to be
asociated with mortality. On the other hand, those
fish exposed to 48 ppm cadmium for 96 hr all died
within a few days after being returned to clean
seawater (Calabrese, Collier, and Miller this report,
Part I). Therefore, as regards the effects of cadmium
on the cunner, mortality seems to be associated with
severe pathological changes of an apparently irreversi-
ble nature taking place in the kidney.
Epidermis
The destruction of the mucus cells of the epidermis
could have important consequences as it would
eliminate the fish’s first line of defense against infec-
tious microorganisms. The possibility that heavy
metals would destroy mucus cells with exposures too
short to cause renal damage should be investigated. In
the present study, the fish had already received a
Figures 7 and 8.—Blood smear of control fish (a) and fish exposed to 48 ppm cadmium for 96 hr (b). Note poikilocytosis, karyolkasis,
and abundant smudge cells in 7b, 8b. 185 (7), 750X (8); Wright’s stain.
lethal exposure to the metal so that destruction of the
mucus cells was not an important factor in prognosis.
Blood
An understanding of the changes taking place in the
blood of the cunner must await further study. The in-
crease in eosinophils noted by Gardner and Yevich
(1970) was not seen, possibly because of the longer ex-
posures used in this study. Wilson et al. (1941) noted
an increase of eosinophils of rats after dietary ex-
posure to cadmium, and Friberg (1950) found an in-
crease of eosinophils in rabbit blood from a normal 3%
level to 25% after exposure to cadmium oxide dust.
Thrombocytopenia, lymphocytopenia, and
neutrophilia in the present study parallel the changes
found by Gardner and Yevich (1970) in mummichogs
and have also been found in other species under stress
or after adrenal corticoid administration (Weinreb,
1958).
32
LITERATURE CITED
AHLMARK, A., B. AXELSSON, L. FRIBERG, and M. PISCATOR.
1961. Further investigations into kidney function and pro-
teinuria in chronic cadmium poisoning. Proc. 13th Int.
Congr. Occup. Health, p. 201-203.
BILINSKI, E., and R. E. E. JONAS.
1973. Effects of cadmium and copper on the oxidation of lac-
tate by rainbow trout (Salmo gairdneri) gills. J. Fish. Res.
Board Can. 30:1553-1558.
BURTON, D. T., E. L. MORGAN, and J. CAIRNS, JR.
1972. Mortality curves of bluegills (Lepomis macrochirus
Rafinesque) simultaneously exposed to temperature and zi
stress. Trans. Am. Fish. Soc. 101:435-441.
CALABRESE, A., R. S. COLLIER, and J. E. MILLER.
1974. Physiological response of the cunner, Tautogolab
adspersus, to cadmium. J. Introduction and experimental
design. Jn Physiological response of the cunner, Tauto-
golabrus adspersus, to cadmium, p. 1-3. NOAA Tech. Rep.
NMES SSRF 681
DALHAMN, T., and L. FRIBERG.
1957. Morphological investigations on kidney damage in
chronic cadmium poisoning. Acta Pathol. Microbiol.
Scand. 40:475-479.
DOUDOROFF, P., and M. KATZ.
1953. Critical review of literature on the toxicity of industrial
wastes and their components to fish. II. The metals, as salts.
Sewage Ind. Wastes 25:802-839.
EISLER, R.
1971. Cadmium poisoning in Fundulus heteroclitus (Pisces:
Cyprinodontidae) and other marine organisms. J. Fish.
Res. Board Can. 28:1225-1234.
FOSTER, C. L., and E. CAMERON.
1963. Observations on the histological effects of sub-lethal
doses of cadmium chloride in the rabbit. II. The effect on the
kidney cortex. J. Anat. 97:281-288.
FRIBERG, L.
1950. Health hazards in the manufacture of alkaline accumu-
lators with special reference to chronic cadmium poisoning.
Acta Med. Scand. Suppl. 240, 124 p.
1955. Iron and liver administration in chronic cadmium
poisoning and studies of the distribution and excretion of
cadmium. Experimental investigations in rabbits. Acta
Pharmacol. 11:168-178.
FRIBERG, L., M. PISCATOR, and G. NORDBERG.
1971. Cadmium in the environment. CRC Press, Cleveland,
166 p.
GARDNER, G. R., and P. P. YEVICH.
1970. Histological and hematological responses of an estu-
arine teleost to cadmium. J. Fish. Res. Board Can.
27:2185-2196.
GOULD, E., and J. J. KAROLUS.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. V. Observations on the biochemistry.
33
In Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium, p. 21-25. NOAA Tech. Rep.
NMFS SSRF 681.
KAGI, J. H. R., and B. L. VALLEE.
1960. Metallothionein: A cadmium- and zinc-containing pro-
tein from equine renal cortex. J. Biol. Chem. 235:3460-3465.
MOUNT, D.I., and C. E. STEPHAN.
1967. A method for detecting cadmium poisoning in fish. J.
Wildl. Manage. 31:168-172.
PISCATOR, M.
1962. Proteinuria in chronic cadmium poisoning. I. An
electrophoretic and chemical study of urinary and serum
proteins from workers with chronic cadmium poisoning.
Arch. Environ. Health 4:607-621.
SKIDMORE, J. F.
1964. Toxicity of zinc compounds to aquatic animals, with
special reference to fish. Q. Rev. Biol. 39:227-248.
1970. Respiration and osmoregulation in rainbow trout with
gills damaged by zinc sulphate. J. Exp. Biol. 52:481-494.
THURBERG, F. P., and M. A. DAWSON.
1974. Physiological response of the cunner, Tautogolabrus
adspersus, to cadmium. III. Changes in osmoregulation and
oxygen consumption. Jn Physiological response of the
cunner, Tautogolabrus adspersus, to cadmium, p. 11-13.
NOAA Tech. Rep. NMFS SSRF 681.
WEINREB, E. L.
1958. Studies on the histology and histopathology of the rain-
bow trout, Salmo gairdneri irideus. 1. Hematology: under
normal and experimental conditions of inflamation. Zoo-
logica 43:145-153.
WILSON, R. H., F. DeEDS, and A. J. COX.
1941. Effects of continued cadmium feeding. J. Pharm-
acol. Exp. Ther. 71:222.
% U.S. GOVERNMENT PRINTING OFFICE: 1974—697- 788/i9 REGION I0
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648. Weight loss of pond-raised channel catfish (Ictalurus punctatus) during holding in
processing plant vats. By Donald C. Greenland and Robert L. Gill. December 1971, iii + 7
pp., 3 figs., 2 tables. For sale by the Superintendent of Documents, U.S. Government
Printing Office, Washington, D.C. 20402.
649. Distribution of forage of skipjack tuna (Euthynnus pelamis) in the eastern tropical
Pacific. By Maurice Blackburn and Michael Laurs. January 1972, iii + 16 pp., 7 figs., 3
tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
650. Effects of some antioxidants and EDTA on the development of rancidity in Spanish
mackerel (Scomberomorus maculatus) during frozen storage. By Robert N. Farragut.
February 1972, iv + 12 pp., 6 figs., 12 tables. For sale by the Superintendent of
Documents, U.S. Government Printing Office, Washington, D.C. 20402.
651. The effect of premortem stress, holding temperatures, and freezing on the
biochemistry and quality of skipjack tuna. By Ladell Crawford. April 1972, iii + 23 pp., 3
figs.. 4 tables. For sale by the Superintendent of Documents, U.S. Government Printing
Office, Washington, D.C. 20402.
653. The use of electricity in conjunction with a 12.5-meter (Headrope) Gulf-of-Mexico
shrimp trawl in Lake Michigan. By James E. Ellis. March 1972, iv + 10 pp., 11 figs., 4
tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
654. An electric detector system for recovering internally tagged menhaden, genus
Brevoortia. By R. O. Parker, Jr. February 1972, iii + 7 pp., 3 figs., 1 appendix table. For
sale by the Superintendent of Documents, U.S. Government Printing Office, Washington,
D.C. 20402. ;
655. Immobilization of fingerling salmon and trout by decompression. By Doyle F.
Sutherland. March 1972, iii + 7 pp., 3 figs., 2 tables. For sale by the Superintendent of
Documents, U.S. Government Printing Office, Washington, D.C. 20402.
656. The calico scallop, Argopecten gibbus. By Donald M. Allen and T. J. Costello. May
1972, iii + 19 pp., 9 figs., 1 table. For sale by the Superintendent of Documents, U.S.
Government Printing Office, Washington, D.C. 20402.
657. Making fish protein concentrates by enzymatic hydrolysis. A status report on
research and some processes and products studied by NMFS. By Malcolm B. Hale.
November 1972, v + 32 pp., 15 figs., 17 tables, 1 appendix table. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402.
658. List of fishes of Alaska and adjacent waters with a guide to some of their literature.
By Jay C. Quast and Elizabeth L. Hall. July 1972, iv + 47 pp. For sale by the Superinten-
dent of Documents, U.S. Government Printing Office, Washington, D.C. 20402.
659. The Southeast Fisheries Center bionumeric code. Part I: Fishes. By Harvey R.
Bullis, Jr., Richard B. Roe, and Judith C. Gatlin. July 1972, xl + 95 pp., 2 figs. For sale by
the Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402.
660. A freshwater fish electro-motivator (FFEM)-its characteristics and operation. By
James E. Ellis and Charles C. Hoopes. November 1972, iii + 11 pp., 9 figs.
661. A review of the literature on the development of skipjack tuna fisheries in the cen-
tral and western Pacific Ocean. By Frank J. Hester and Tamio Otsu. January 1973, iii +
13 pp., 1 fig. For sale by the Superintendent of Documents, U.S. Government Printing Of-
fice, Washington, D.C. 20402.
662. Seasonal distribution of tunas and billfishes in the Atlantic. By John P. Wise ana
Charles W. Davis. January 1973, iv + 24 pp., 13 figs., 4 tables. For sale by the Superinten-
dent of Documents, U.S. Government Printing Office, Washington, D.C. 20402.
663. Fish larvae collected from the northeastern Pacific Ocean and Puget Sound during
April and May 1967. By Kenneth D. Waldron. December 1972, iii + 16 pp., 2 figs., 1 table,
4 appendix tables. For sale by the Superintendent of Documents, U.S. Government Print-
ing Office, Washington, D.C. 20402.
664. Tagging and tag-recovery experiments with Atlantic menhaden, Brevoortia tyran-
nus. By Richard L. Kroger and Robert L. Dryfoos. December 1972, iv + 11 pp., 4 figs., 12
tables. For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
665. Larval fish survey of Humbolt Bay, California. By Maxwell B. Eldridge and Charles
F. Bryan. December 1972, iii + 8 pp., 8 figs., 1 table. For sale by the Superintendent of
Documents, U.S. Government Printing Office, Washington, D.C. 20402.
666. Distribution and relative abundance of fishes in Newport River, North Carolina. By
William R. Turner and George N. Johnson. September 1973, iv + 23 pp., 1 fig., 13 tables.
For sale by the Superintendent of Documents, U.S. Government Printing Office,
Washington, D.C. 20402.
667. An analysis of the commercial lobster (Homarus americanus) fishery along the coast
of Maine, August 1966 through December 1970. By James C. Thomas. June 1973, v + 57
pp., 18 figs., 11 tables. For sale by the Superintendent of Documents, U.S. Government
Printing Office, Washington, D.C. 20402.
668. An annotated bibliography of the cunner, Tautogolabrus adspersus (Walbaum). By
Fredric M. Serchuk and David W. Frame. May 1973, ii + 43 pp. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
20402.
669. Subpoint prediction for direct readout meteorological satellites. By L. E. Eber.
August 1973, iii + 7 pp., 2 figs., 1 table. For sale by the Superintendent of Documents,
U.S. Government Printing Office, Washington, D.C. 20402.
670. Unharvested fishes in the U.S. commercial fishery of western Lake Erie in 1969. By
Harry D. Van Meter. July 1973, iii + 11 pp., 6 figs., 6 tables. For sale by the Superinten-
dent of Documents, U.S. Government Printing Office, Washington, D.C. 20402.
671. Coastal upwelling indices, west coast of North America, 1946-71. By Andrew
Bakun. June 1973, iv + 103 pp., 6 figs., 3 tables, 45 appendix figs. For sale by the
Superintendent of Documents, U.S. Government Printing Office, Washington, D.C.
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672. Seasonal occurrence of young Gulf menhaden and other fishes in a northwestern
Florida estuary. By Marlin E. Tagatz and E. Peter H. Wilkins. August 1973, iii + 14 pp., 1
fig., 4 tables. For sale by the Superintendent of Documents, U.S. Government Printing Of-
fice, Washington, D.C. 20402.
673, Abundance and distribution of inshore benthic fauna off southwestern Long Island,
N.Y. By Frank W. Steimle, Jr. and Richard B. Stone. December 1973, iii + 50 pp., 2 figs.,
5 appendix tables.
674. Lake Erie bottom trawl explorations, 1962-66. By Edgar W. Bowman. January 1974,
iv + 21 pp., 9 figs., 1 table, 7 appendix tables.
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