JOURNALOFTHE
1
WORLD AQUACULTURE SOCIETY
Effects of Low Salinities on Oxygen Consumption of Selected
Euryhaline and Stenohaline Freshwater Fish
AND JOHNM. GRIZZLE^
ILHAN ALTINOK
Southeastern Cooperative Fish Disease Project, Department of Fisheries and Allied
Aquacultures, Auburn University, Auburn, Alabama 36849 USA
The amount of energy required for os-
moregulation depends on the difference be-
tween internal and external concentrations
of ions (Rao 1968; Farmer and Beamish
1969), changes in corticosteroid hormone
levels (Morgan and Iwama 1996), glomer-
ular filtration rates (Furspan et al. 1984),
gill and kidney Na+, K+-ATPase activity
(McCormick et al. 1989; Morgan and Iwa-
ma 1998), tissue permeability to water and
ions, and gill ventilation, perfusion, and
functional surface area (Rankin and Bolis
1984). Differences in the energetic cost of
osmoregulation play a significant role in the
difference in growth rate between seawater-
and freshwater-adapted fish (Morgan and
Iwama 1991; Ron et al. 1995; Wang et al.
1997). Oxygen consumption is an indirect
indicator of metabolic rate in fish (Cech
1990) and can be used to determine effects
of salinity changes on energy costs.
Most researchers agree that salinities dif-
ferent from that of the internal fluids of the
fish must impose energetic regulatory costs
for active ion transport (Morgan and Iwama
1998). However, there is less agreement
concerning the magnitude of these costs
(Rao 1968; Farmer and Beamish 1969;
Nordlie and Leffler 1975; Nordlie 1978;
Furspan et al. 1984; Febry and Lutz 1987;
Morgan and Iwama 1991; Claireaux and
Lagard&re 1999) and for most species there
is little information about the energetic con-
sequences of life in different salinities.
Our objective was to investigate the ef-
fects of low salinities (59.Woo) on oxygen
consumption in experiments allowing com-
parison of phylogenetically diverse species.
I Corresponding author.
63 Copyright by
Vol. 34, No.
March, 2003
The salinity range was selected to include
approximately isosmotic and hypo-osmotic
conditions. With the conditions we used,
the oxygen measurements provided an es-
timate of the resting routine metabolic rate,
which requires fasted, quiescent fish (Cech
1990). To eliminate metabolic requirements
associated with acclimation, fish were kept
in test salinities for 2.5 mo before oxygen
consumption was determined.
Six species of stenohaline and euryhaline
fishes were tested. Channel catfish Ictalurus
punctatus and goldfish Carassius auratus
are stenohaline fish species that can only
tolerate less than half-strength sea water
(Black 1951; Allen and Avault 1970).
However, rainbow trout Oncorhynchus my-
kiss, brown trout Salmo trutta, striped bass
Morone saxatilis, and Gulf sturgeon Aci-
penser oxyrinchus desotoi (a subspecies of
Atlantic sturgeon A. oxyrinchus) are eury-
haline species that can live in a wide range
of salinities from fresh water to full-
strength sea water depending on their size
and life stage (Behnke 1992; Elliott 1994;
Zehfuss et al. 1999; Secor et al. 2000).
Materials and Methods
Source and Maintenance of Fish
Fish were held in recirculating systems
with different salinities for 2.5 mo until
used for respirometry. At the time of res-
pirometry, channel catfish (Marion X Ar-
kansas strain) were 6.5 mo old and weighed
6.7 2 1.1 g, goldfish were 7 mo old and
weighed 7.0 2 1.2 g, rainbow trout (Shasta
strain) were 3.5 mo old and weighed 6.3 2
1.0 g, brown trout (Walhalla strain) were 5
mo old and weighed 5.5 t 0.9 g, striped
bass (Savannah River strain) were 5.5 mo
the World Aquaculture Society 2003
113
114 ALTINOK AND GRIZZLE

TABLEI . Water characteristics (mean 2 SD) during the acclimation period in the respiratory chamber. Sa-
linity, conductivity, pH, hardness, and alkalinity were checked daily, and dissolved oxygen was checked f o r
every fish. Ammonia and nitrite were not detected. For dissolved oxygen and conductivity, the first line is f o r
salmonids and the second line is f o r other species.

Salinity
Characteristics Fresh water 1.o%o 3.o%o 9.o%o
Salinity (%o) 0.0 t 0.0 1.0 t 0.1 3.0 5 0.1 9.0 t 0.1
Conductivity (pS/cm) 89 t9 1,811 t 45 4,890 +. 112 13,190 t 103
98 t 7 2,180 t 32 5,610 2 92 15,250 t 104
Dissolved oxygen 8.9 t 0.2 8.9 t 0.1 8.9 t 0.1 8.8 t 0.2
(mgn) 7.7 t 0.5 7.7 t 0.2 7.7 t 0.2 7.6 t 0.2
PH 7.3 2 0.2 7.4 t 0.2 7.5 2 0.3 7.6 t 0.2
Total hardness
(mg/L as CaCO,) 22 ? 2 208 t 14 561 t 24 1,446 t 32
Total alkalinity
( m a as CaCO,) 21 5 1 31 + . 3 42 ? 3 54 t 4

old and weighed 9.4 2 1.6 g, and Gulf stur-
geon were 6 mo old and weighed 13.4 2
1.6 g.
%ice a day, fish were fed with Zeigler
salmon starter (Zeigler Brothers, Gardners,
Pennsylvania, USA) totaling 5% body
weight/d. Fish were subsampled every
week to adjust feeding rates. Photoperiod
was 12 h light: 12 h dark. Waste was si-
phoned from the aquaria daily.
Water Quality
Water of different salinities (Table 1) was
prepared by mixing well water and artificial
sea salts (Instant Ocean Synthetic Seasalt,
Mentor, Ohio, USA) and waiting at least 24
h prior to use. Salinity was measured with
a conductivity-salinity meter (Yellow
Springs Instruments, Yellow Springs, Ohio,
USA). During the respiratory experiment,
water temperature was 23.5 2 0.7 C (mean
2 SD) for warmwater species, 16.3 2 0.5
C for rainbow trout, and 17.2 2 0.7 C for
brown trout. Hardness, alkalinity, ammonia,
nitrite, conductivity, and salinity were
checked daily during the acclimation period
in the chamber, and temperature was re-
corded prior to oxygen consumption mea-
surement for every fish.
Oxygen Consumption
Respiratory experiments were conducted
in two 2.1-L and one 5.7-L, transparent, cy-
lindrical plastic chambers. For fasting, an
individual fish was moved from a recircu-
lating system to a 30-L aquarium contain-
ing static aerated water with the same sa-
linity as in the recirculating system. After
24 h in the holding aquarium, the fish was
moved to a respiratory chamber 12 h before
the experiment. Respiratory chambers were
covered with black plastic throughout ac-
climation (12 h) and testing periods (1 h).
During the acclimation period, the respira-
tory chamber received a continuous supply
(6-7 L/h) of aerated water. Fish were not
fed for 36 h prior to oxygen measurement.
To determine the oxygen consumption,
water flow was stopped and after 15 min,
dissolved oxygen concentration was record-
ed every 3 to 10 min for 1 h. A polaro-
graphic oxygen meter and thermistor (Mod-
el 95; Yellow Springs Instruments) were
used to measure dissolved oxygen and tem-
perature in the chamber. Six replicate trials
were conducted per salinity for each spe-
cies. Each fish was weighed and measured
for total length after oxygen consumption
measurements. All experiments were be-
tween 0800 and 1700 h. To account for bac-
terial consumption of oxygen in the system,
a blank trial without fish was run after each
fish.
The rate of change in dissolved oxygen
concentration (pg/L) during the 1-h mea-
 EFFECTS OF SALINITY ON OXYGEN CONSUMPTION 115

TABLE2. Oxygen consumption (wg O,/g per h ) of stenohaline and euryhaline fish in different salinities. In
each row. means with a common letter ure not significantly different (P > 0.05). Six replicate trials were
conducted per salinity for each species. Water temperature was 23.5 2 0.7 C (mean _C SD) for warmwater
species. 16.3 2 0.5 C for rainbow trout, and 17.2 5 0.7 C,for brown trout.

Salinity
Pooled
Fish species Fresh water I .o%o 3.0%0 9.o%o SEM
Channel catfish 251 z 271 z 221 z 239 z 18
Goldfish 138 y I30 y 134 y 213 z 12
Rainbow trout 252 x 221 y 148 z 137 z 4.7
Brown trout 223 z 186 y 161 y 226 z 10
Striped bass 259 y 261 y 233 yz 211 z I1
Gulf sturgeon 300 y 365 z 358 z 352 z 13

surement period was determined with linear Results and Discussion

regression (average 9 = 0.934). Fish oxy- The salinities tested had no effect on
gen consumption was calculated with the channel catfish oxygen consumption while
following formula: goldfish oxygen consumption was 54-64%
Oxygen consumption (pg O,/g per h) higher in 9.0%0salinity than in other treat-
= (sf - sb).V/W ments (Table 2). Oxygen consumption of
rainbow trout and striped bass decreased
where sf is the regression coefficient with a with increasing salinity. For brown trout,
fish in the chamber, sb is the regression co- oxygen consumption was lowest in 1.0 and
efficient without a fish in the chamber, V is 3.0%0 salinities and was highest in fresh
volume of the respiratory chamber (L), and water and 9.0%0salinity. For Gulf sturgeon,
W is the fish mass (g). oxygen consumption was 15-18% lower in
After each experiment, the respiratory fresh water than in any other salinity. Al-
chamber was disinfected with chlorine and though Gulf sturgeon were larger than the
then rinsed for 30 min. Initial dissolved other species tested, oxygen consumption
oxygen concentration in the respiratory per gram was significantly higher for Gulf
chambers was 8.65-9.02 mg/L for cold- sturgeon than for other species in any salin-
water species and 7.82-8.04 mg/L for ity.
warmwater species. Minimum ending dis- Except for rainbow trout, the relation be-
solved oxygen concentration was 7.53-7.96 tween salinity and oxygen consumption has
mg/L for coldwater species and 5.80-6.64 not been previously considered for the spe-
mg/L for warmwater species. cies used in the present study. The results
of the present study agree with Rao (1968),
Statistical Analyses who found that oxygen consumption of
Oxygen consumption (pg O,/g per h) for rainbow trout was minimum in isosmotic
each species and among species was eval- water regardless of activity level. However,
uated by ANOVA using SAS Version 6 Morgan and Iwama (1991) found that met-
software (SAS Institute, Inc., Cary, North abolic rate of rainbow trout with a swim-
Carolina, USA). For each species, the ming speed of 1 body length/sec was higher
pooled SEM was calculated from the AN- in 9%0salinity than in fresh water, but that
OVA as follows: SEM = (MSE/N)o.5,where there were no significant differences in met-
MSE is the mean square error. When sig- abolic rates of steelhead 0. mykiss in salin-
nificant differences were found (P < 0.05), ities from 0 to 12%0. The difference be-
comparisons of means were made with tween the two groups of 0. mykiss (rainbow
Fisher's LSD test. trout and steelhead) used by Morgan and
116 ALTINOK AND GRIZZLE
Iwama ( I 99 1) highlights the genetic vari-
ability within this species, which could ex-
plain the apparent discrepancies among
studies with this species.
In an earlier study, the specific growth
rate (SGR) was reduced for channel catfish
and goldfish but increased for rainbow trout
and striped bass as salinity increased to 9%0
(Altinok and Grizzle 2001). Alteration of
the energetic cost of osmoregulation in dif-
ferent salinities was a potential explanation
for these changes in SGR. The results of
the present study support this hypothesis for
rainbow trout, striped bass, and goldfish.
Results for brown trout further support this
hypothesis; the highest SGR was in 3%0 sa-
linity (Altinok and Grizzle 2001) and the
lowest rate of oxygen consumption was at
the same salinity. However, the similar ox-
ygen consumption of channel catfish in dif-
ferent salinities and the minimum oxygen
consumption of Gulf sturgeon in fresh wa-
ter, while SGR was higher in 3%0 (Altinok
and Grizzle 2001), indicate that these spe-
cies have a more complex relation between
oxygen consumption and the energy re-
quired for osmoregulation.
The differences we found in oxygen con-
sumption in different salinities could result
from differences in energy required for os-
moregulation or from changes in the spon-
taneous activity level of fish. Salinity can
affect the amount of spontaneous fish activ-
ity (Job 1969), and thereby change the rou-
tine metabolic rate. The amount of fish ac-
tivity can also affect the cost of osmoreg-
ulation (Febry and Lutz 1987). The respi-
ratory chambers were covered during the
present study to minimize disturbance of
the fish, but the level of activity was not
monitored. Additional research is needed to
determine to what extent the differences in
oxygen consumption of fish in different sa-
linities are influenced by changes in the
amount of spontaneous activity.
Isosmotic water did not reduce resting
routine metabolic rates for the sizes of
channel catfish, goldfish, brown trout, and
Gulf sturgeon in the present study. Oxygen
consumption was minimum in isosmotic
water (9.Wo0) for only rainbow trout and
striped bass. It is likely that the metabolic
response to salinity in some fish species is
dependent on the developmental stage of
osmoregulatory ability (Parry 1958; Job
1969; Nordlie 1978). Variation among the
species used in the present study probably
reflects differences in their natural history,
degree of euryhalinity, and developmental
stage related to osmoregulatory capabilities.
Acknowledgments
This study was supported by the South-
eastern Cooperative Fish Disease Project.
We thank the Georgia Department of Nat-
ural Resources for providing rainbow trout,
brown trout, and striped bass; T. B. Henry
for providing channel catfish; Blue Spring
Fish Hatchery for providing goldfish; and
E A. Chapman for providing Gulf sturgeon.
We also thank M. S. West and J. A. Os-
borne for their advice on statistical analysis.
Literature Cited
Allen, K. 0. and J. W. Avault, Jr. 1970. Effects of
salinity on growth and survival of channel catfish,
Ictalurus punctatus. Proceedings of the South-
eastern Association of Game and Fish Commis-
sioners 23(1969):319-331.
Altinok, I. and J. M. Grizzle. 2001. Effects of brack-
ish water on growth, feed conversion and energy
absorption efficiency by juvenile euryhaline and
freshwater stenohaline fishes. Journal of Fish Bi-
ology 59:1142-1152.
Behnke, R. J. 1992. Native trout of Western North
America. American Fisheries Society, Bethesda,
Maryland, USA.
Black, V. S. 195I . Osmotic regulations in teleost fish-
es. Pages 53-89 in W. S. Hoar, V. S. Black, and
E. C. Black, editors. Some aspects of the physi-
ology of fish. University of Toronto Press, Toron-
to, Canada.
Cech, J. J., Jr. 1990. Respirometry. Page 335-362 in
C. B. Schreck and P. B. Moyle, editors. Methods
for fish biology. American Fisheries Society, Be-
thesda, Maryland, USA.
Claireaux, G. and J.-P. Lagardhe. 1999. Influence
of temperature, oxygen and salinity on the metab-
olism of the European sea bass. Journal of Sea
Research 42:157-168.
Elliott, J. M. 1994. Quantitative ecology and the
 EFFECTS O F SALINITY ON OXYGEN CONSUMPTION
brown trout. Oxford University Press, Oxford,
UK.
Farmer, G. J and F. W. H. Beamish. 1969. Oxygen
consumption of Tilapia nilorica in relation to
swimming speed and salinity. Journal of the Fish-
eries Research Board of Canada 26:2807-2821.
Febry, R. and P. Lutz. 1987. Energy partitioning in
fish: the activity-related cost of osmoregulation in
a euryhaline cichlid. Journal of Experimental Bi-
ology 128:63-85.
Furspan, P., H. D. Prange, and L. Greenwald. 1984.
Energetics and osmoregulation in the catfish, Ic-
ralurus nehulosus and 1. punctatus. Comparative
Biochemistry and Physiology 77A:773-778.
Job, S. V. 1969. The respiratory metabolism of Tilapia
mossumhica (Teleostei): I. The effect of size, tem-
perature and salinity. Marine Biology 2: 121-126.
McCormick, S. D., C. D. Moyes, and J. S. Ballan-
tyne. 1989. Influence of salinity on the energetics
of gill and kidney of Atlantic salmon (Salmo sa-
lar). Fish Physiology and Biochemistry 6:243-
2.54.
Morgan, J. D. and G. K. Iwama. 1991. Effects of
salinity on growth, metabolism, and ion regulation
in juvenile rainbow and steelhead trout (Oncor-
hynchus mykiss) and fall chinook salmon (Oncor-
hynchus tshu wytscha). Canadian Journal of Fish-
eries and Aquatic Sciences 48:2083-2094.
Morgan, J. D. and G. K. Iwama. 1996. Cortisol-in-
duced changes in oxygen consumption and ionic
regulation in coastal cutthroat trout (Oncorhyn-
chus clurki clurki) pan: Fish Physiology and Bio-
chemistry 15:385-394.
Morgan, J. D. and G. K. Iwama. 1998. Salinity ef-
fects on oxygen consumption, gill Na', K+-
ATPase and ion regulation in juvenile coho salm-
on. Journal of Fish Biology 53: 1 1 10-1 119.
Nordlie, F. G. 1978. The influence of environmental
117
salinity on respiratory oxygen demands in the eu-
ryhaline teleost, Ambussis interruptu Bleeker.
Comparative Biochemistry and Physiology 59A:
271-274.
Nordlie, F. G. and C. W. LefRer. 1975. Ionic regu-
lation and the energetics of osmoregulation in Mu-
gil cephalus Lin. Comparative Biochemistry and
Physiology 5 I A: 125-1 3 1.
Parry, G. 1958. Size and osmoregulation in salmonid
fishes. Nature 181: 1218-1219.
Rankin, J. C. and L. Bolis. 1984. Hormonal control
of water movement across the gills. Pages 177-
201 in W. S. Hoar and D. J. Randall, editors. Fish
physiology, volume I OB. Academic Press, Orlan-
do, Florida, USA.
Rao, G. M. M. 1968. Oxygen consumption of rainbow
trout (Salmo gairdneri) in relation to activity and
salinity. Canadian Journal of Zoology 46:78 1-
786.
Ron, B., S. K. Shimoda, G. K. Iwama, and E. G.
Grau. 1995. Relationships among ration, salinity,
17-a methyltestosterone and growth in the eury-
haline tilapia. Oreochrornis mossambicus. Aqua-
culture 135:185-193.
Secor, D. H., T. E. Gunderson, and K. Karlsson.
2000. Effect of temperature and salinity on growth
performance in anadromous (Chesapeake Bay)
and nonanadromous (Santee-Cooper) strains of
striped bass Morone suxatilis. Copeia 2000:291-
296.
Wang, J. Q., H. Lui, H. Po, and L. Fan. 1997. In-
fluence of salinity on food consumption, growth
and energy conversion efficiency of common carp
(Cyprinus carpio) fingerlings. Aquaculture 148:
115-124.
Zehfuss, K. P., J. E. Hightower, and K. H. Pollock.
1999. Abundance of Gulf sturgeon in the Apa-
lachicola River, Florida. Transactions of the
American Fisheries Society 128: 130-143.