EFFECTS OF TEMPERATURE ON CHRONIC HYPOXIA TOLERANCE

IN THE NON-INDIGENOUS BROWN MUSSEL, PERNA PERNA

(BIVALVIA: MYTILIDAE) FROM THE TEXAS GULF OF MEXICO
DAVID W. HICKS 1 AND ROBERT F. M C MAHON 2
1
Department of Biological Sciences, The University of Texas at Brownsville, 80 Fort Brown, Brownsville, Texas 78520, USA;
2
Department of Biology/Honors College, Box 19222, The University of Texas at Arlington, Arlington, Texas 76019, USA
(Received 26 January 2004; accepted 31 March 2005)

ABSTRACT

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Effects of temperature (158, 208 and 258C), O2 partial pressure (PO2 ¼ 0, 1, 2, 4, and 6 kPa), and indi-
vidual size (12 – 79 mm shell length; SL) on survivorship of specimens of the non-indigenous, marine,
brown mussel, Perna perna, from Texas were investigated to assess its potential distribution in North
America. Its hypoxia tolerance was temperature-dependent, survivorship being significantly extended
at lower temperatures under all tested lethal PO2. Incipient tolerated PO2 was
4 and
6 kPa at 15 and
208C, respectively, with .50% mortality occurring at 258C at all tested levels of hypoxia. PO2 had less of
an effect on survival of hypoxia than temperature. At 258C, survivorship was not different over a PO2
range of 0 – 2 kPa and increased only at 4 and 6 kPa. Survivorship was size-dependent. Median survival
times increased with increasing SL in anoxia and PO2 ¼ 1 kPa, but at 2, 4 and 6 kPa, smaller individuals
survived longer than larger individuals. With tolerance levels similar to other estuarine bivalve species,
P. perna should withstand hypoxia encountered in estuarine environments. Thus, its restriction to inter-
tidal rocky shores may be due to other parameters, particularly its relatively low temperature tolerance.

INTRODUCTION (Marshall & McQuaid, 1993). Although Marshall &

The subtropical, intertidal, non-indigenous, marine mussel,
Perna perna (Linnaeus, 1758), was introduced to the Gulf of
Mexico in the late 1980s where it was first discovered in 1990
on jetty rocks in Port Aransas, Texas (Hicks & Tunnell, 1993).
The potential distribution of P. perna in North American
marine and estuarine waters is unknown. Since its discovery, it
has invaded new areas along the Texas and Mexico Gulf of
Mexico coasts, including several estuarine habitats in Texas
(Davenport, 1995; Hicks & Tunnell, 1995; McGrath, Hyde &
Tunnell, 1998). Shallow bays and estuaries with poor water cir-
culation are subject to periodic hypoxia (i.e. PO2 below air O2
saturation levels of 21 kPa) as a result of biological oxygen
demand by decomposing organic matter under conditions of
temperature and/or salinity stratification, and stagnation or
eutrophication (reviewed by Diaz & Rosenberg, 1995). While
open water, rocky-shore habitats are typically normoxic, sessile
intertidal bivalves may also experience periodic hypoxia as a
result of prolonged valve closure in response to tidal emersion
(Moon & Pritchard, 1970; McMahon, 1988), salinity extremes
(Bayne, 1973; Salomão, Magalhães, & Lunetta, 1980), or sand
burial (Berry, 1978; Marshall & McQuaid, 1993).
Tolerance of prolonged hypoxia (.5 days) has rarely been
tested in marine molluscs. Studies have been limited to the inter-
tidal neogastropod, Stramonita haemastoma (Kapper & Stickle,
1987; Stickle et al., 1989; Das & Stickle, 1993), and the marine
bivalves, Crassostrea virginica (Stickle et al., 1989), Mytilus gallopro-
vincialis, Scapharca inaequivalvis (de Zwaan et al., 1991), Mytilus
edulis (Theede et al., 1969) and Perna perna (Marshall &
McQuaid, 1993). South African specimens of Perna perna with-
stand sand burial for at least 10 – 12 days ( Jackson, 1976;
Berry, 1978), while laboratory studies indicated that it has an
LT50 of  6.5 days at 35‰ salinity and 208C in hypoxic
seawater at PO2 1.5– 3 kPa (7 – 13% of full air O2 saturation)
Correspondence: D. W. Hicks; e-mail: david.hicks@utb.edu
McQuaid (1993) provided some data on the hypoxia tolerance
of South African P. perna, its chronic hypoxia tolerance (i.e. inci-
pient limits), and the effects of temperature and individual size
on chronic hypoxia tolerance, have not been empirically
assessed. This paper describes an investigation of chronic
hypoxia tolerance in P. perna within its typical ambient tempera-
ture range of 15 – 258C (Carvajal, 1969; Jackson, 1976), in order
to predict its potential to colonize North American marine and
estuarine coastal habitats.
MATERIAL AND METHODS
Specimens of Perna perna were collected from intertidal rocks on
the north jetty of Mansfield Pass (268340 N) on the Texas Gulf of
Mexico coast and transported overnight in cooled, insulated
containers to Arlington, Texas. Upon arrival, mussels were
maintained in a 284-l aerated holding tank containing 35‰ sali-
nity artificial seawater (created with Fritz Supersalt and City of
Arlington, Texas, dechlorinated tapwater) at a constant tempe-
rature of 208C on a 12 :12 h light:dark cycle without feeding
prior to experimentation. Specimens of P. perna can survive
under laboratory conditions without food for periods far exceed-
ing the experimental treatments utilized in this study (e.g. 6
months at 208C; personal observation). All experiments were
initiated within 30 days of collection.
The chronic hypoxia tolerance of P. perna was examined at PO2
values of 0, 1, 2, 4 and 6 kPa (0%, 5%, 10%, 20% and 30% of
full air O2 saturation, respectively) at temperatures of 158, 208
and 258C. This temperature range was within the 10– 308C inci-
pient limits previously determined for Gulf of Mexico popu-
lations of this species (Hicks & McMahon, 2002a). Individuals
(shell lengths, SL ¼ 12– 80 mm) were excised from mussel
clumps by cutting their byssal attachment threads with scissors.
Groups of four to five excised mussels, of similar size range, were
placed into crystallization dishes (5 cm height by 9 cm diameter)
in order to prevent reattachment to each other in large clumps

Journal of Molluscan Studies (2005) 71: 401– 408. Advance Access Publication: 5 August 2005 doi:10.1093/mollus/eyi042
# The Author 2005. Published by Oxford University Studies on behalf of The Malacological Society of London, all rights reserved.
 D. W. HICKS AND R. F. M C MAHON
and to facilitate subsequent viability determinations. Groups of
four dishes were randomly assigned to 4-l plastic aquaria (i.e.
16 mussels/container) with tightly fitting, vented lids. Each
aquarium was filled with 3-l of seawater, leaving a 1-l head
space. Three replicate 4-l aquaria were then randomly assigned
to each temperature/PO2 treatment combination, while three
similar 4-l aquaria served as controls. Hypoxic conditions were
maintained by bubbling media with appropriate mixtures of
O2 and N2 maintained by a Cameron Instruments Gas Mixing
Flowmeter (Model GF-3/MP). Control aquaria were continu-
ously aerated with atmospheric air (PO2  21 kPa). Oxygen
partial pressures in test and control media were monitored
daily with a YSI Model 53, Clarke-type oxygen electrode.
Prior to experimentation, individuals were habituated to
test temperatures of 158, 208 and 258C under normoxic con-
ditions for 14 days, during which they byssally reattached to
the walls of holding dishes and shells of other mussels. The
aquaria were then placed in an environmental chamber held
at the appropriate test temperature, and the hypoxia and
control treatments initiated. Media were changed in experi-
mental and control containers daily to avoid accumulation
of potentially toxic metabolites. Replacement media were
held at the appropriate test temperature and PO2 before
being added to test containers. Shell blackening, resulting
from precipitation of microbially generated sulphides and
metals under anaerobic conditions (de Zwaan et al., 2001),
was not observed in the anoxia treatment. Thus, anoxia tole-
rance measurements were unlikely to have been impacted by
release of sulphide and other noxious metabolites by anaerobic
bacteria (de Zwaan et al., 2001).
Viability, byssal attachment, and valve gaping behaviour of
all individuals were examined at 24-h intervals. Viability was
tested by prodding mantle edges of immersed individuals with
the bristles of a fine brush. Individuals not closing their valves
in response to this stimulus were considered dead, removed
from the container, measured (linear distance from the anterior
to posterior margins of the shell to the nearest 0.1 mm using dial
calipers) and the time interval recorded at which death was
observed. Prior observations indicated that non-responsive,
gaping individuals did not recover on return to normoxic con-
ditions. Exposure to hypoxic-temperature treatments was main-
tained until either 100% mortality was achieved or individuals
survived an exposure of 30 days.
The hypoxia tolerance of specimens of P. perna was examined
using the discrete logistic failure time model (DLFTM) (Cox,
1972; Hicks, Hawkins & McMahon, 2000). This approach
allowed determination of the effects of temperature and PO2 on
survival duration in the presence of censored observations (e.g.
some individuals survive treatments), using only categorized sur-
vival times (i.e. monitoring at 24-h intervals), while controlling
for individual-specific covariates (e.g. size as SL) (see Hicks et al.,
2000 for details of DLFTM used in this study).
Which variables (e.g. SL, or treatment-by-SL interactions) to
include in the DLFTM analysis were determined by beginning
with a model deemed a priori general enough (e.g. at most, a
quadratic dependence on SL and SL-by-treatment interactions)
and testing a sequence of nested models until the most parsimo-
nious model to adequately explain the data was reached.
Goodness-of-fit in each tested survival model was assessed by
comparing the most parsimonious fitted model to the a priori
model (i.e. the model containing, at most, quadratic functions
of SL and treatment-by-SL interaction) by a Wald statistic
(Wald, 1943). Treatment combinations of PO2 and test tempera-
ture were included in the model as indicator variables utilizing a
reference cell parameterization (e.g. PO2 ¼ 0 and 258C as the
arbitrarily assigned reference cell). Survival probability curves
were generated from the survival probabilities estimated by
the model at each 24-h interval endpoint.
402
Median survival times were estimated by linear interpolation
across the time interval for which death occurred for those treat-
ments in which survival probability fell below 0.5 (Hicks et al.,
2000). However, survival probabilities in several treatments
never fell below 0.5; consequently, median survival times could
not be estimated. Therefore, post-hoc statistical testing among
treatment pairs was limited to SL-adjusted pairwise comparisons
of odds-of-death ratios, and not based on median survival times
(Hicks et al., 2000). Exponentiation of treatment regression coef-
ficients (relating the log-odds of death in any interval to the
reference cell treatment) provided a corresponding factor by
which the odds of death relate to the reference cell treatment
(i.e. an odds ratio). Exponentiated treatment coefficient values
equal to 1 indicate the same odds of death as the reference cell
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treatment, values significantly . 1 indicate an increase in the
odds of death relative to the reference cell treatment, and
values significantly ,1 indicate a decrease in the odds of death
relative to the reference cell treatment. The odds ratios are
simply related to survival probabilities in that if the odds of
death are statistically greater in one treatment relative to
another, then survival probability in any time interval is signifi-
cantly lower.
Shell-length adjusted median survival time estimates and
their standard errors (SE) were provided for all treatments
where possible (i.e. where survival probabilities fell below 0.5).
The DLFTM analysis was implemented in a computer
program written in SAS’s interactive matrix language (IML,
SASw, Cary, North Carolina) (Hicks et al., 2000).
RESULTS
The most parsimonious, fitted model relating survival time of
specimens of Perna perna to temperature, PO2 and individual
size included linear functions of SL and SL-by-treatment inter-
actions (i.e. Ho: SL2, SL2 by treatment ¼ 0; x2[15] ¼ 19.235,
P ¼ 0.2033), specifically

 X
15 X15
1  pj x
log ¼ b1 þ b TI T þ g T I T ðSL  SLo Þ
pi x
T¼2 T¼2
þ g1 ðSL  SLo Þ
where b and g are coefficients for treatment and SL, respect-
ively, SLo is the specified value of SL and IT is the indicator
variable for treatment (i.e. IT ¼ 1 for treatment T and 0
otherwise). Each PO2/test temperature combination was given
a unique indicator (i.e. 1 – 15, the PO2 ¼ 0 kPa/258 test tempera-
ture treatment was designated as the reference cell).
The estimated survival curves for the various PO2/test temp-
erature treatment combinations, adjusted to the sample mean
SL (42 mm), indicated decreasing survival probabilities with
increasing temperature and decreasing PO2 (Fig. 1, Table 1).
No mortality of individuals of P. perna occurred in fully aerobic
control treatments (PO2 ¼ 21 kPa). Similarly, there was almost
no mortality recorded at 158 and 208C under the highest
tested PO2 (6 kPa). PO2/test temperature combinations of 6 kPa
and 158C, and 6 kPa and 208C appeared non-lethal to this
species, as 96% of individuals in these treatments survived the
30-day (720-h) exposure period (Table 1). Survival duration
was also affected by SL (x2[15] ¼ 397.93, P , 0.0001). Median
survival times estimated at the 10th, 25th, 50th, 75th and 90th
percentiles of the individual SL distribution (i.e. 24, 29, 38, 58
and 65 mm SL, respectively) indicated a general trend of
increasing survival time with increasing SL in both the anoxia
and the lowest PO2 treatment of 1 kPa. At treatments of

2 kPa the trend was reversed, with smaller individuals surviv-
ing longer relative to larger individuals (Fig. 2).
 HYPOXIA TOLERANCE IN PERNA PERNA

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Figure 1. Survival probability curves for standard 42 mm shell length Texas specimens of Perna perna on chronic exposure to PO2 of (A) 0, (B) 1, (C) 2,
(D) 4 and (E) 6 kPa at test temperatures of 158, 208 and 258C. Error bars represent one standard error of survival probability. Note: a median survival
time is the hour at which the survival probability curve crosses the solid horizontal line at a survival probability of 0.5.

At a PO2 of 0 kPa and 158C, 23% of specimens survived the 30-
day trial, whereas complete mortality was recorded at this PO2
after 684 and 276 h among the 208 and 258C treatments, respec-
tively (Fig. 1A). The odds of death (in any time interval) for
specimens in the 158C anoxic treatment (i.e. PO2 ¼ 0 kPa)
were only 2.6% of the odds of death for specimens experiencing
anoxia at 258C (P , 0.001) (Table 2). Median survival time for
an averaged sized individual (SL ¼ 42 mm) in the anoxic treat-
ment at 158C (587.3 h) was more than four times greater than
that recorded at 258C (135.4 h) (Table 1). The odds of death
were also significantly reduced in the 208C anoxic treatment
compared with that of the 258C anoxic treatment (e.g. 15.1%,
P , 0.001), as the median survival time at 208C (290.4 h) was
more than twice that at 258C (135.4 h) (Table 1). In contrast,
the factor by which the odds of death differed between the 158
and 208C anoxic treatments was not significantly different
(P ¼ 0.087) (Table 3).
A similar pattern of temperature dependence was observed in
the PO2 ¼ 1 kPa treatment in which the odds of death were sig-
nificantly reduced at 158C relative to that at 258C (P , 0.001)
(Table 3). Eight percent of individuals held under a PO2 of
1 kPa at 158C survived 30 days. Complete mortality was
recorded at 540 and 324 h at PO2 ¼ 1 kPa and 208 and 258C,
respectively (Fig. 1B). The median survival time for specimens
held at 158C (354.0 h) exceeded that for specimens held at
258C by 2.5 times (137.1 h) (Table 1). Survivorship of indivi-
duals in PO2 ¼ 1 kPa at 208C (median ¼ 222.4 h) also exceeded
that of individuals at 258C (P , 0.001), but was significantly
reduced compared with those held at 158C (P ¼ 0.050)
(Table 3). Survivorship at PO2 ¼ 1 kPa for the 158, 208C and

403
 D. W. HICKS AND R. F. M C MAHON

Table 1. Median survival time and time to total sample mortality on treatment, and at 6 kPa over the 1 kPa treatment (P ¼ 0.002)
exposure to chronic hypoxia (PO2 ¼ 0, 1, 2, 4 and 6 kPa) for specimens (Table 1, 3). The same survival pattern was observed for specimens
of Perna perna (n ¼ number of specimens) adjusted to a shell length held at 158C and 208C, in which survival did not improve over the
(SL) of 42 mm at test temperatures of 158, 208 and 258C. PO2 ¼ 0–2 kPa treatments with survival times significantly increas-
ing only at PO2 treatments 52 kPa (P , 0.05) (Tables 1, 3).
Temperature PO2 Median survival Time of 100% n SL range
Increasing PO2, rather than temperature, appeared to have a
(kPa) time [h (+ SE)] mortality (h) (mm) greater effect on the behaviour of P. perna specimens in experi-
mental treatments. The percentage of individuals with open
158C 0 587.3 (58.66) .720 48 18.4 –62.6
shell valves was similar among all temperatures at the same PO2
1 354.0 (19.45) .720 51 30.1 –67.4
level. For example, in the anoxic treatment (PO2 ¼ 0 kPa), the
2 564.6 (129.5) .720 48 14.7 –74.0 average percentage of individuals with open valves over the
4 .720 .720 48 11.9 –79.2 course of survival was 87.6%, 83.3% and 73.6% in the 158, 208
6 .720 .720 48 49.8 –79.3 and 258C treatments, respectively. Similarly, the respective per-
centages of individuals with open valves were 86.6%, 79.9% and

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208C 0 290.4 (16.94) 684 57 17.6 –61.0
79.2%, and 87.1%, 89.3% and 93.7% at 158, 208 and 258C in the
1 222.4 (31.55) 540 53 20.3 –64.4 PO2 ¼ 1 and 2 kPa treatments, respectively. In contrast, among
2 346.8 (23.35) .720 48 14.7 –74.0 the higher PO2 treatments of 4 kPa and 6 kPa, the percentages
4 424.4 (49.52) .720 48 16.0 –73.5 of individuals with open valves were markedly lower, being
6 .720 .720 48 51.0 –72.7 27.6%, 29.5% and 33.3%, and 33.8%, 54.3% and 41.3%
among the 158, 208 and 258C treatments, respectively.
258C 0 135.4 (8.71) 276 51 18.4 –62.6 Specimens of P. perna did not abandon their byssal holdfasts in
1 137.1 (9.02) 324 51 17.5 –62.9 response to hypoxic stress at any tested level of hypoxia or temp-
2 156.6 (22.5) 324 48 21.8 –54.8 erature. Indeed, fewer than 5% of specimens detached during
4 244.3 (19.8) .720 46 19.0 –70.4 any monitoring period. Most of those that did detach usually
6 538.3 (137.87) .720 35 45.3 –75.6 reattached prior to the next 24-h monitoring interval.

258C treatments did not differ from that recorded at the same
temperatures in the anoxic treatments (P . 0.05) (Table 3).
In the PO2 ¼ 2 kPa treatment, survivorship was again signifi-
cantly reduced for specimens held at 258C relative to those held
at either 158 or 208C (P , 0.05) (Table 3). Sixty-two percent of
individuals held under PO2 ¼ 2 kPa at 158C survived 30 days,
compared with 13% at 208C, with complete mortality occurring
after 300 h among individuals held at 258C (Fig. 1C). The factor
by which the odds of death differed among the 158 and 208C
PO2 ¼ 2 kPa treatments was not significantly different
(P ¼ 0.993) (Table 3). Median survival times for the 158, 208
and 258C 2 kPa-PO2 treatments were 564.6, 346.8 and 156.5 h,
respectively (Table 1).
In the PO2 ¼ 4 kPa treatment, survivorship was significantly
reduced for specimens held at 258C relative to those held at
158 or 208C (P , 0.05) (Table 3). Fifty-one percent of indivi-
duals held under PO2 ¼ 4 kPa at 158C survived 30 days, compared
with 47% survival among those held at 208C and 14% survival
among those held at 258C (Fig. 1D). Survivorship among speci-
mens held at 208C and 258C was not significantly different
(P ¼ 0.197) (Table 3). Median survival times for the 208C
and 258C, PO2 ¼ 4 kPa treatments were 424.4 and 244.3 h
respectively (Table 1). A median survival time could not be esti-
mated for specimens held at 158C since survival probabilities
never fell below 0.5 during the 30-day observation period
(Fig. 1D).
High survival (.50% of sample) in the PO2 ¼ 6 kPa treat-
ments precluded accurate estimation of survival parameters,
especially at 158 and 208C in which only 4% mortality was
recorded after the 30-day observation period. However, a
median survival time of 538 h was estimated for specimens
held at 258C (Table 1) in which 17% of specimens survived
the 30-day observation period (Fig. 1E).
The level of PO2 had less of an effect on the hypoxia tolerance of
P. perna than did temperature (Table 1, Fig. 1). For example,
survivorship did not improve significantly at 258C over PO2 levels
of 0–2 kPa (median survival times ¼ 135.4, 137.1 and 156.5 h at
PO2 ¼ 0, 1 and 2 kPa, respectively) (Tables 1, 3). Survival at
258C did improve significantly in the PO2 ¼ 4 (P ¼ 0.029) and
6 kPa (P ¼ 0.002) treatments over that recorded for the anoxic
DISCUSSION
Hypoxia tolerance times determined for Perna perna in this study
were similar to those of other marine and estuarine bivalves.
South African specimens of P. perna had an LT50 of 6.5 days
under PO2  1.5– 3 kPa (7 – 13% of full air O2 saturation) at
35‰ salinity and 208C (Marshall & McQuaid, 1993), a result
consistent with the estimated 9-day median survival time
recorded for Texas specimens at a similar PO2 (i.e. 1 kPa) and
test temperature (208C). The estuarine mytilid, Mytilus edulis,
had an LT50 of 35 days under  0.5 kPa at 108C (Theede
et al., 1969) whereas Mytilus galloprovincialis survived anoxia up
to 18 days in 36‰ salinity at 208C with an LT50 of 15.1 days
(de Zwaan et al., 1991). At a salinity of 30‰, the estuarine
oyster, Crassostrea virginica, had LT50 values of . 28 days at a
PO2 of 0 kPa and 108C, 20 days at a PO2 of 4 kPa and 208C,
and 3 days at a PO2 of 16 kPa and 308C (Stickle et al., 1989).
The infaunal bivalve species, Arctica islandica, Scrobicularia plana,
Mya arenaria and Cerastoderma edule, had LT50 values of 55, 25,
21 and 4 days, respectively, on exposure to PO2  0.5 kPa at
108C (Theede et al., 1969). Scapharca inaequivalvis survived
anoxia up to 22 days in 36‰ salinity at 208C with an LT50 of
18.3 days (de Zwaan et al., 1991). In contrast, the infaunal
species, Mulinia lateralis, had lower survival times under anoxic
conditions, LT50 values being 5.5 days at 108C, 4 –7 days
at 208C and 2 days at 308C (Shumway & Scott, 1983). The
corresponding median survival times for Texas specimens of P.
perna were comparable with those cited above for other intertidal
and estuarine infaunal bivalve species.
The hypoxia tolerance of P. perna was temperature dependent.
Survivorship significantly improved at low temperatures under
all tested lethal O2 partial pressures. For example, specimens
at 158C in the anoxic treatment survived significantly longer
than those held at 258C under PO2 ¼ 4 kPa (20% of full air O2
saturation) (P ¼ 0.002; Tables 1, 3). Ambient temperatures
approaching lethal limits in marine bivalves have been shown
to induce a transition to anaerobiosis even in fully aerated sea-
water as a result of a breakdown of the oxygen supply mecha-
nism (Pörtner, 2001, 2002; Peck, Pörtner & Hardewig, 2002).
However, since the 15 – 258C temperature range utilized in this
study was well within the long-term temperature limits (10 –
308C) of P. perna (Hicks & McMahon, 2002a), it is unlikely

404
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Figure 2. Median survival times (h) for Texas specimens of Perna perna on chronic exposure to PO2 of (A) 0, (B) 1, (C) 2, (D) 4 and (E) 6 kPa at test
temperatures of 158, 208 and 258C, adjusted to the 10th (24 mm SL, open histograms), 25th (29 mm SL, left-hatched histograms), 50th (38 mm SL,
cross-hatched histograms), 75th (58 mm SL, right-hatched histograms) and 90th (65 mm SL, shaded histograms) percentiles of the individual SL
distribution.

that the poorer survival observed at elevated temperatures
involved a temperature-induced reduction in gill ventilation
and capacity to transport O2 to internal tissues. Rather, the
trend of decreasing survivorship with increasing temperature
in P. perna was consistent with this species’ rather limited
capacity to regulate its O2 consumption rate in increasing
ambient temperatures
. (Hicks & McMahon, 2002b). The O2
consumption rate (VO2) of Texas specimens of P. perna increases
with increasing temperature and is thermally
. regulated only
between 158 and 208C. Beyond 208C, VO2 increases sharply,
with Q10 values in warm acclimated specimens (258C) of 2.83
and 1.65 over 20 – 258C and 25 – 308C, respectively (Hicks &
McMahon, 2002b). At lower . temperatures, metabolic
demands are reduced, with VO2 at 108C being only 25% of
that at 258C. High respiratory Q10 values above 208C indicate
that metabolic demands dramatically increase in the upper tol-
erated temperature range of P. perna. Thus, mismatches between
environmental O2 concentration and metabolic demand will
have greater consequences at high temperatures, accelerating
the development of energy deficits, anaerobiosis, and eventual
collapse of physiological function. Furthermore, low tissue O2
levels and transition to anaerobiosis results in elevated pro-
duction of potentially lethal anaerobic end-products which
accumulate more rapidly at higher ambient temperatures
(Pörtner et al., 1999; Peck et al., 2002), potentially leading to
the reduced survivorship observed at elevated temperatures in
this study.

405
 D. W. HICKS AND R. F. M C MAHON

Table 2. Exponentiated treatment coefficients relating the odds of death uptake with progressive hypoxia is consistent with their
on exposure to five different chronic hypoxia treatments of specimens of increased tolerance of hypoxic treatments greater than
Perna perna, adjusted to a shell length of 42 mm at test temperatures of PO2 ¼ 1 kPa and could be attributable to their increased
158, 208 and 258C, to the reference cell treatment (PO2 ¼ 0 kPa at 258C). surface area to volume ratios relative to larger individuals. Diffu-
sion of O2 from epithelial surfaces to internal tissues is an import-
Temperature PO2 (kPa)
ant O2 transport mechanism in intertidal mytilids. Diffusion of
0 1 2 4 6
O2 accounted for 85% of O2 transport in the mytilid, Geukensia
demissus, with only 15% being accounted for by haemolymph cir-
158C 0.0262 0.0885 0.0292 0.0156 0.0033 culation (Booth & Mangum, 1978). Thus, the larger surface
208C 0.1510 0.2900 0.0947 0.0577 0.0001 area to volume ratios and reduced diffusive distances of
258C 1 0.9625 0.6853 0.3203 0.0324 smaller specimens of P. perna may support greater O2 uptake
and transport rates, accounting for their increased survival at
The value in each cell represents the factor by which the odds of death PO2 between 1 and 6 kPa. The reduced tissue surface to
increase/decrease relative to the reference cell. Any two treatments can be volume ratios and increased body thickness of larger individuals

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compared by the ratio of their odds of death (e.g. the odds of death under
PO 2 ¼ 0 kPa at 158C divided by the odds of death under PO 2 ¼ 6 kPa at
258C equals 0.8, thus, the odds of death are similar (1) in these two
treatments; the odds of death for all possible temperature/P O 2 treatment
combinations are displayed in Table 3). When such treatment coefficient
ratios are greater than 7 or less than 0.15 they are generally significant at
P , 0.05.
Capacity to regulate O2 uptake with progressive hypoxia in
Texas specimens of P. perna increased with increasing tempera-
ture and decreasing individual size (Hicks & McMahon, 2002b).
The increased capacity of smaller specimens to regulate O2
are likely to reduce their capacity for diffusive uptake of O2 in
hypoxic conditions, leading to increased reliance on anaerobiosis
and accumulation of deleterious anaerobic metabolites (Pörtner
et al., 1999). Increasing the surface area of mantle tissues exposed
to the media in order to facilitate diffusion of O2 could account
for the extensive gaping behaviour observed in individuals in the
most hypoxic treatments (i.e. PO2 ¼ 0, 1 and 2 kPa). This result
is consistent with the observation that South African specimens
of P. perna exhibited valve gaping behaviour when exposed to
hypoxic seawater (PO2  1.5 – 3 kPa) (Marshall & McQuaid,
1993).

Table 3. Ratios of odds with corresponding P values in parentheses (x2[14]) from a Scheffés test of pairwise differences in treatment coefficients relating
the odds of death on exposure to five different chronic hypoxia treatments of specimens of Perna perna (PO2 in kPa) adjusted to a shell length of 42 mm at
test temperatures of 158, 208 and 258C.

6 kPa 4 kPa 2 kPa 1 kPa 0 kPa

PO2 Temp. 258 208 158 258 208 158 258 208 158 258 208 158 258 208
0 158 0.81 195.89 8.01 0.10 0.45 1.68 0.04 0.28 0.99 0.03 0.09 0.30 0.03 5.76
(0.999) (0.998) (0.999) (0.002) (0.995) (0.999) (,0.001) (0.744) (0.999) (,0.001) (,0.001) (0.729) (,0.001) (0.087)
208 4.65 1128.2 46.12 0.60 2.61 9.69 0.22 1.59 5.16 0.16 0.52 1.69 0.15
(0.919) (0.976) (0.999) (0.995) (0.546) (,0.001) (0.307) (0.999) (0.828) (,0.001) (0.916) (0.989) (,0.001)
258 0.03 0.001 0.003 0.25 0.06 0.02 0.69 0.09 0.03 0.96 0.29 0.09
(0.002) (0.835) (0.974) (0.029) (,0.001) (,0.001) (0.999) (,0.001) (,0.001) (0.999) (0.065) (,0.001)
1 158 2.72 662.46 27.07 0.35 1.53 5.69 0.13 0.93 3.03 10.87 3.29
(0.999) (0.989) (0.999) (0.302) (0.999) (,0.001) (0.011) (0.999) (0.995) (,0.001) (0.050)
208 8.95 2170.5 88.70 1.15 5.03 18.64 0.42 3.06 9.93 3.32
(0.385) (0.948) (0.998) (0.999) (,0.001) (,0.001) (0.979) (0.297) (0.227) (0.100)
258 29.69 7203.3 294.4 3.82 16.69 61.87 1.40 10.16 32.94
(0.002) (0.840) (0.978) (0.061) (,0.001) (,0.001) (0.999) (,0.001) (,0.001)
2 158 0.90 218.66 8.94 0.12 1.88 0.51 23.45 3.24
(0.999) (0.999) (0.999) (0.362) (0.999) (0.999) (0.029) (0.993)
208 2.92 708.87 28.97 0.38 1.64 6.09 7.23
(0.996) (0.988) (0.999) (0.676) (0.999) (0.003) (0.045)
258 21.14 5128.4 209.59 2.72 11.88 44.05
(0.066) (0.882) (0.987) (0.941) (,0.001) (,0.001)
4 158 0.48 116.43 4.76 16.21 3.71
(0.999) (0.999) (0.999) (,0.001) (0.197)
208 1.78 431.66 17.64 4.37
(0.999) (0.995) (0.999) (0.013)
258 7.78 1887.3 77.13
(0.538) (0.955) (0.998)
6 158 9.91 0.04
(0.999) (0.999)
208 242.60
(0.988)


Indicates significance difference: P , 0.05.

406
 HYPOXIA TOLERANCE IN PERNA PERNA
In the lowest PO2 treatments of 0 and 1 kPa, larger specimens
survived significantly longer than smaller individuals. Larger
individuals have proportionally greater organic energy stores
and lower mass-specific metabolic rates than smaller individuals
(Hicks & McMahon, 2002b). Thus, under complete anoxia or
extreme hypoxia, cellular energetic requirements could be sup-
ported by anaerobic metabolism over a longer period in larger
individuals than in more metabolically active smaller indivi-
duals with reduced energy stores. In addition, the higher
mass-specific metabolic rates of smaller specimens (Hicks &
McMahon, 2002b) could cause accumulation of anaerobic
end-products to lethal levels more rapidly than in larger
specimens.
To date, P. perna has only ephemerally colonized estuarine
areas in Texas. Surveys of Texas bay environments recorded
adult specimens of P. perna at Port O’Connor in March and
June 1996, and Corpus Christi Bay, the Brownsville Ship
Channel and the lower Laguna Madre in June – August 1995
(McGrath et al., 1998). When these sites were re-surveyed, speci-
mens were no longer present at Port O’Connor in September
1996, or Corpus Christi Bay and the Brownsville Ship Channel
in October 1996. When last surveyed in March 1997, specimens
of P. perna were still present in the lower Laguna Madre;
however, the population was in decline (McGrath et al., 1998).
The ephemeral presence of P. perna in Texas estuaries indicates
that conditions in these estuaries may be periodically conducive
to this species’ settlement and growth, but cannot indefinitely
sustain populations. Thus, the apparent extirpation of several
established Texas estuarine P. perna populations suggests that
this species is not fully adapted to the range of ambient con-
ditions characteristic of these habitats.
Throughout its worldwide distribution, Perna perna is primar-
ily found on exposed, wave-swept, rocky shores ( Jackson, 1976;
Berry, 1978). Like other intertidal organisms it can be subjected
to short-term hypoxia during tidal emersion (Diaz & Rosenberg,
1995). Exposure to hypoxia during tidal emersion may have
allowed evolution of hypoxia tolerance in P. perna that is
similar to that of truly estuarine species (see above), suggesting
that this species should be capable of colonizing estuarine habi-
tats. Byers (2000) has shown that introduced, hypoxia-tolerant,
estuarine gastropods can displace less tolerant native species
under anthropogenic euthrophication. However, because
P. perna is restricted primarily to open-water, marine intertidal
habitats in Texas (Hicks & Tunnell, 1995) and throughout its
worldwide range (Berry, 1978), hypoxia tolerance may not be
the primary factor involved in its apparent exclusion from estu-
arine habitats. Similarly, the incipient salinity tolerance of
P. perna, at 15 – 50‰ salinity, does not preclude it from estuarine
waters (Hicks et al., 2000).
The relatively low incipient upper lethal temperatures for
P. perna embryonic development, veliger larvae, and adults at
258, 308 and 308C, respectively (Romero & Moreira 1980;
Hicks & McMahon, 2002a) are more likely to prevent its suc-
cessful colonization of estuarine habitats within its subtropical
range. Ambient water temperatures rarely exceed 308C in
open water, subtropical, marine habitats, but often exceed
358C in subtropical, estuarine habitats (Galtsoff, 1964; Lane,
1986) and approach 308C in temperate estuarine habitats
(Kennedy & Mihursky, 1971; Ansell et al., 1981; Wilson,
1991). Thus, periodic exposures to ambient temperatures
approaching or exceeding the 308C incipient upper thermal
limit of P. perna are likely to preclude its colonization of estuaries.
Indeed, intertidal estuarine bivalves have distinctly higher
temperature tolerances than do rocky shore bivalves (Bayne,
Thompson & Widdows, 1976; Wright et al., 1983; Rajagopal,
Nair & Azariah, 1995). Temperature as the primary factor
excluding P. perna from estuarine habitats, is supported by the
reported collapse of Gulf of Mexico populations during 1997
407
and 1998 when average summer water temperatures reached
or exceeded 308C for the first time since this species discovery
on the Texas Gulf coast in 1990 (Hicks & McMahon, 2002a).
ACKNOWLEDGEMENTS
Ronald Smith and Terry Riggs of Texas A&M University-
Corpus Christi collected and arranged transport of specimens
of Perna perna to Arlington, Texas. Jennifer Shaw of the Univer-
sity of Texas at Arlington assisted in the laboratory.
Dr D. L. Hawkins of the University of Texas at Arlington
provided valuable assistance with statistical analysis of the
data. This research was funded by a grant from the Texas
Downloaded from https://academic.oup.com/mollus/article-abstract/71/4/401/1002935 by guest on 16 February 2020
A&M Sea Grant College Program (Grant #NA56RG0388).
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