Marine Pollution Bulletin 73 (2013) 428–434

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Marine Pollution Bulletin

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Effects of elevated CO2 on the reproduction of two calanoid copepods

Kristian McConville a,c, Claudia Halsband a,b,⇑, Elaine S. Fileman a, Paul J. Somerfield a, Helen S. Findlay a,
John I. Spicer c
a
Plymouth Marine Laboratory, Prospect Place, The Hoe, Plymouth PL1 3DH, UK
b
Akvaplan-niva AS, Fram Centre, N-9296 Tromsø, Norway
c
Marine Biology and Ecology Research Centre, School of Marine Science and Engineering, University of Plymouth, Plymouth, Devon PL4 8AA, UK

a r t i c l e i n f o a b s t r a c t

Keywords: Some planktonic groups suffer negative effects from ocean acidification (OA), although copepods might
Carbon capture be less sensitive. We investigated the effect of predicted CO2 levels (range 480–750 ppm), on egg produc-
Centropages tion and hatching success of two copepod species, Centropages typicus and Temora longicornis. In these
Ocean acidification short-term incubations there was no significant effect of high CO2 on these parameters. Additionally a
Reproduction
very high CO2 treatment, (CO2 = 9830 ppm), representative of carbon capture and storage scenarios,
Temora
resulted in a reduction of egg production rate and hatching success of C. typicus, but not T. longicornis.
In conclusion, reproduction of C. typicus was more sensitive to acute elevated seawater CO2 than that
of T. longicornis, but neither species was affected by exposure to CO2 levels predicted for the year
2100. The duration and seasonal timing of exposures to high pCO2, however, might have a significant
effect on the reproduction success of calanoid copepods.
Ó 2013 Elsevier Ltd. All rights reserved.

1. Introduction Copepods often constitute the majority of the biomass of zoo-

When atmospheric CO2 dissolves into the oceans, the pH level is
lowered, a phenomenon called ocean acidification (OA) (Riebesell
et al., 2010). Average surface seawater pH is predicted to decrease
from pH 8.2 to pH 7.6 by 2100 (Caldeira and Wickett, 2003; IPCC
2007). This constitutes an increase in surface ocean acidity one
hundred times faster than any known natural change over several
million years (Riebesell et al., 2010). Potential effects of ocean acid-
ification, such as lowered pH and hypercapnia (increased [CO2]) on
marine organisms are beginning to be elucidated (Fabry et al.
2008), but often the results are not straightforward.
Plankton are important in determining the health of the oceans
(Hays et al., 2005). Studies have demonstrated that planktonic
organisms (in particular those bearing calcareous shells) exhibit
chronic effects in response to increases in pCO2. For example, the
holoplanktonic molluscs, Clio pyrimidata (L.) and Limacina helicina
(Phipps) show shell dissolution and decreased calcification, respec-
tively, over relatively short exposures to elevated pCO2 seawater of
790 and 760 ppm CO2 respectively (Orr et al. 2005; Comeau et al.
2009). Furthermore, planktonic larvae of Crassostrea gigas (Thun-
berg) underwent increases in deformation of veliger shells and
Ophiothrix fragilis (Abilgaard in O.F. Müller) in larval mortality
(Kurihara et al., 2007; Dupont et al. 2008).
⇑ Corresponding author at: Akvaplan-niva AS, Fram Centre, N-9296 Tromsø,
Norway. Tel.: +47 77750347.
E-mail address: claudia.halsband@akvaplan.niva.no (C. Halsband).
plankton and play a vital role in marine ecosystems, serving as cru-
cial links between primary production and higher trophic levels.
Despite their ecological significance the potential effects of ocean
acidification on copepods remain unclear as few studies have
examined CO2 conditions realistic by the year 2100. Mayor et al.
(2012) exposed Calanus finmarchicus to 1000 ppm CO2, but found
no significant effect on offspring viability. However, an interactive
positive effect with temperature, i.e. higher viability with elevated
temperature, was observed. Most previous studies investigating ef-
fects of ocean acidification on copepods have used CO2 concentra-
tions beyond those expected in the next 100 years by the IPCC, but
are relevant as they emulate potential Carbon Capture & Storage
(CCS) conditions (e.g. Kurihara et al. 2004; Mayor et al. 2007; Kuri-
hara and Ishimatsu 2008). The term CCS represents a number of
methods by which anthropogenically-generated CO2 is collected
at source and released into mesopelagic waters, seafloor depres-
sions or geological sub-surface formations (Liro et al., 1992; Parson
and Keith, 1998). Estimates suggest that CO2 concentrations of
20,000 ppm and pH levels as low as 5.8 are likely near the point
of release, if a leak were to occur (Herzog et al., 1996).
Potential impacts of CCS on marine zooplankton are summa-
rized in Halsband and Kurihara (2013). Kurihara et al. (2004)
observed a significant decrease in egg production but no significant
effect on adult mortality in the copepod Acartia steueri in response
to CO2 concentrations of >10,000 ppm. In a more recent study,
Kurihara and Ishimatsu (2008) exposed Acartia tsuensis to CO2
concentrations of 2830 ppm, a potential value for the year 2300

0025-326X/$ - see front matter Ó 2013 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.marpolbul.2013.02.010
 K. McConville et al. / Marine Pollution Bulletin 73 (2013) 428–434 429

(Caldeira and Wickett, 2003), but also possible in the waters surround- Alk2) following Dickson et al. (2007), and using certified reference
ing a CCS leak, and observed that survival, body size and develop- materials from Dickson (Batch 100, TA = 2242.4).
ment speed remained unaffected. In their study, inter-generational
effects were also considered, demonstrating that egg production
3. Experiments
and hatching success remained unaffected over two generations.
Mayor et al. (2007), however, observed a reduction in hatching
Four experiments over 4 days each were performed with C. typ-
success of Calanus finmarchicus in response to 8000 ppm CO2, dem-
icus (weeks 26–28 and week 31) and three experiments with T. lon-
onstrating that the results may differ between species.
gicornis (weeks 29–31), respectively. It was not possible to run all
While adult copepods may be relatively resilient to increased
treatments for both species during all experiments due to insuffi-
CO2 concentrations, similar conditions may exhibit negative effects
cient numbers of female copepods, but a total of 887 and 386 fe-
in fitness indices such as egg production and juvenile growth, espe-
males of C. typicus and T. longicornis, respectively, were incubated
cially in response to CCS. Here, we investigated the effects of CO2-
across all experiments, with a minimum of 47 individuals at
acidified sea water on egg production and hatching success of two
480 ppm (T. longicornis) and a maximum of 186 individuals at
calanoid copepods, Centropages typicus (Krøyer) and Temora longi-
9830 ppm (C. typicus). Triplicate 250 ml Duran bottles per pH level
cornis (Müller, O.F.). These species differ markedly from Acartia tsu-
were filled with 250 ml sea water at the designated pH levels and
ensis and Calanus finmarchicus in their biogeography, biology and
groups of 2–5 adult female copepods and incubated at an in situ
ecology. Both species are widely distributed throughout the Atlan-
temperature of 14.7 °C. The heterotrophic dinoflagellate, Oxyrrhis
tic Ocean, where T. longicornis, although restricted to coastal re-
marina Dujardin, was added to each bottle at a concentration of
gions, extends into high latitudes (Halsband-Lenk et al., 2002;
approximately 10,000 cells l1 to provide food for the copepods.
Razouls et al., 2005–2011), while C. typicus prefers coastal to open
Every 24 h the contents of each bottle were filtered to retrieve both
ocean habitats in mid-latitudes including the Mediterranean. These
copepods and their eggs. Eggs were washed into petri dishes for
species were also chosen due to their high egg production rates,
counting under a stereo microscope and then transferred into
ease of local collection and taxonomic distance (family Centropag-
15 ml test tubes where they were incubated in their respective
idae and family Temoridae, respectively, Martin and Davis, 2001).
food solutions for 48 h, a period that allowed the eggs to hatch at
The CO2 treatments investigated ranged from present day condi-
ambient temperatures.
tions to those expected for the year 2100 (385, 480, 620,
The experiments were designed such that the copepods experi-
750 ppm CO2) and one concentration representative of an exposure
enced the physiological stress of immediate transition from normal
in the proximity of a CCS leak (9830 ppm CO2). Although Centro-
seawater to seawater with an elevated CO2 concentration. This
pages and Temora are epipelagic species and as such unlikely to
sudden change is potentially representative of a CO2-gradient fol-
encounter CCS pollution, we use them as representatives of their
lowing a CCS leak. Although this design is not directly representa-
taxonomic and functional group that are (1) easily accessible,(2)
tive of CO2-induced ocean acidification, if unaffected by the greater
well suited for experimental manipulation – in contrast to deep-
physiological stress of a sudden change, copepods are even less
sea species – and (3) for which a wealth of ecological information,
likely to be affected by a gradual change over long periods.
especially on reproductive rates, is available.
Egg production rates were determined by counting the number of
eggs spawned during 24 h and dividing by the number of copepods
2. Materials and methods present. Hatching success (after 48 h) was determined by subtract-
ing the number of unhatched eggs from the total number of eggs pro-
2.1. Sampling duced, then dividing by the total number of eggs produced.

Adult female Centropages typicus and Temora longicornis were
collected in vertical net hauls from 50 m depth to the surface with
a WP2 plankton net (mesh size 200 lm) from the L4 sampling site
(50° 15.000 N 4° 13.020 W) in the western English Channel
(www.westernchannelobservatory.org.uk). Sampling was con-
ducted between 28 June and 2 August 2010, when egg production
rates were expected to be high for these species (Halsband and Hir-
che, 2001). All samples were taken between 09:00 and 11:00 local
time.
2.2. Preparation of acidified seawater
Treated seawater was prepared by bubbling CO2 gas into
0.22 lm filtered seawater in 20 l carboys and diluting these sup-
plies with untreated filtered seawater until the nominal pH values
of 8.04, 7.79, 7.85, 7.78 and 6.71 were reached. These correspond to
CO2 concentrations reflecting values expected between the present
day and the year 2100 (Dickson et al., 2007): 385 ppm, 480 ppm,
620 ppm and 750 ppm. A pH of 6.71 represents the nominal CO2
concentration of 10,000 ppm (actual 9830 ppm), chosen to repli-
cate acidified conditions in the water column around a CCS leak
(Herzog et al., 1996). The pH was measured using a pH meter (Met-
tler Toledo pH-/Cond-/DO-/ISE meter) calibrated with Mettler To-
ledo potassium hydroxide buffers. The pH, total alkalinity,
temperature and salinity were measured (Table 1). Alkalinity was
analysed using pentiometric titration (Apollo SciTech Model AS-
3.1. Statistical analyses
Egg production rates and hatching success were analysed using
the PERMANOVA+ add-in (Anderson et al., 2008) for PRIMER v6
(Clarke and Gorley, 2006). PERMANOVA is a standard ANOVA per-
formed here on single variables using Euclidean distance as the
resemblance measure. Significance is determined by permutations,
thereby avoiding normal-distribution theory. For each variable dis-
persion was analysed using PERMDISP, which showed that only
egg production rate had significantly different dispersion across
treatments. To minimise the effect of this variation in dispersion,
egg production rate was transformed (log(x + 1)) prior to conducting
permutational analyses of variance. As measurements were taken on
a daily basis within each week, day was nested within week in the
PERMANOVA design. Also, given the uncertainty as to when each fe-
male last mated (and therefore uncertainty of how much sperm was
stored), day was designated as a random factor. Permutational anal-
yses of variance were run twice per variable for each species, both
including and excluding the more extreme CCS treatment.
4. Results
4.1. Experimental conditions
Total alkalinity did not vary significantly across the different
CO2 treatments (Table 1). The relatively high alkalinity
430 K. McConville et al. / Marine Pollution Bulletin 73 (2013) 428–434

Table 1
Seawater conditions (mean ± standard deviation over 4 days) for each experimental treatment. Salinity, total alkalinity (TA) and pH(NBS) were measured at 14.7 °C. Total inorganic
carbon (TIC), and the saturation states for calcite and aragonite (Xcalcite and Xaragonite) were calculated using CO2sys (Pierrot et al. 2006) with constants from Mehrbach et al.
(1973) refit by Dickson and Millero (1987), and for KSO4 from Dickson (1990).

CO2 Treatment Sal. Temp. (°C) TA (lmol kg1) pH(NBS) TIC (lmol kg1) Xcalcite Xaragonite
380 35.0 ± 0.05 14.7 ± 1.2 2412 ± 23 8.04 ± 0.02 2246 ± 21 3.07 ± 0.12 1.97 ± 0.08
480 35.0 ± 0.05 14.7 ± 1.2 2453 ± 51 7.97 ± 0.02 2310 ± 48 2.79 ± 0.10 1.79 ± 0.06
620 35.0 ± 0.05 14.7 ± 1.2 2417 ± 43 7.85 ± 0.02 2321 ± 42 2.14 ± 0.06 1.38 ± 0.04
750 35.0 ± 0.05 14.7 ± 1.2 2446 ± 57 7.78 ± 0.02 2376 ± 54 1.86 ± 0.10 1.19 ± 0.06
9830 35.0 ± 0.05 14.7 ± 1.2 2455 ± 70 6.71 ± 0.02 2978 ± 83 0.17 ± 0.01 0.11 ± 0.01

(2437 ± 20 across all treatments) maintained the saturation states in situ was around 0.75 in weeks 29 and 30, but high (0.94) in week
of both aragonite and calcite in the ocean acidification scenario 31 (Fig. 1b).
experiments, but in the CCS treatment both saturation states were Hatching success of C. typicus in the experiments was high in all
very low (<0.2). OA treatments with varying degrees of variability between incuba-
tion days and pH treatments. In contrast, egg mortality was high in
4.2. Egg production the CCS incubations; on average less than 60% of the eggs were via-
ble in all three weeks and all but one incubation day (Fig. 3). This
C. typicus egg production rates in situ (Fig. 1a) were near the an- negative effect occurred already after one day of maternal expo-
nual maximum for the species and ranged from over 50– sure, except in week 27, where a large drop in egg viability was ob-
120 eggs per female1 day1, indicating that the females incubated served on day two. Towards the end of the experiments, egg
were in good physiological condition and actively reproducing at mortality appeared less severe, and more than 90% of all eggs
the time of the incubations. hatched on day 4 in experiment 1 (week 26). Hatching success of
Egg production rates (EPRs) showed different trends in the four T. longicornis followed no consistent trend in OA treatments, but
experiments with C. typicus (Fig. 2): in week 26, when in situ egg the CCS treatment had the lowest values on days 1–3 (but not 4)
production was relatively low, the output of eggs per female gen- in week 29 and the 480 ppm treatment was on average lower than
erally increased during the four days of incubation at all CO2 con-
centrations except the CCS treatment. In the latter, EPR were
lowest from outset with 34.5 eggs per female1 day1 and de-
creased further to a minimum of 2.6 eggs per female1 day1 on
day 3. In the following experiments, weeks 27–31, EPR were usu-
ally highest on the first day of incubation and gradually decreased
over the remaining incubation time, independent of pCO2. Females
incubated at the CCS ppm CO2 level spawned the lowest number of
eggs in all three experiments (no data in week 31), decreasing by
63% and 90% in weeks 27 and 28, respectively.
In situ egg production rates of T. longicornis were considerably
lower than those of C. typicus, not exceeding 35 eggs female1
day1 (Fig. 1a). Egg production rates could not be determined at
all pCO2 concentrations in all weeks. All treatments were tested
in week 29, where rates were low (610 eggs female1 day1) and
further decreased over the four days of incubation (Fig. 4). In week
30 controls and lowest OA conditions (380 and 430 ppm) were re-
peated; in week 31 only the CCS concentration (9830 ppm) was
studied again. In these three experiments egg production rates
started around 30 eggs female1 day1 and then decreased over
time. Although the pattern of decreasing egg production was sim-
ilar in all three experiments, lowest egg production rates of
<10 eggs female1 day1 were observed in the CCS incubation,
especially on the last two days of incubation (Fig. 4).
There was no statistically significant effect of pCO2 on egg pro-
duction in any of the OA treatments for both copepods (Table 2).
Upon inclusion of the CCS treatment, a significant effect of pCO2
was observed on the egg production rate of C. typicus (Table 2a),
but not of T. longicornis (Table 2b). Incubation day had a significant
effect on egg production in both OA and CCS treatments and both
copepods. In both OA and CCS treatments and for both species the
p-value for an effect of week upon egg production was 60.079. In
addition, a significant interactive effect was observed between
pCO2 and week in C. typicus, but not T. longicornis.

4.3. Hatching Success

C. typicus hatching success in situ was near maximal in weeks Fig. 1. (a) In situ egg production rates (eggs female1 day1) and (b) hatching
26, 27 and 31, but somewhat decreased to 0.84 in week 28 success (proportion of viable eggs) of C. typicus and T. longicornis from late June
(Fig. 1b). The proportion of viable eggs produced by T. longicornis 2010 (week 26) to early August 2010 (week 31).
 K. McConville et al. / Marine Pollution Bulletin 73 (2013) 428–434 431

200 1,0
385 ppm
480 ppm
620 ppm
750 ppm

egg hatching success

9830 ppm 0,8
150
egg production rate

0,6
100
0,4

50 385 ppm
0,2 480 ppm
620 ppm
750 ppm
9830 ppm

0 0,0

200 day 1,0

egg hatching success

0,8
150
egg production rate

0,6
100
0,4

50
0,2

0 0,0

200
day 1,0
egg hatching success

0,8
150
egg production rate

0,6
100
0,4

50
0,2

0 0,0

200 1,0
egg hatching success

0,8
150
egg production rate

0,6

100
0,4

50
0,2

0,0
0 1 2 3 4
1 2 3 4
day
day
Fig. 3. Effect of elevated CO2 concentrations on the proportion of viable C. typicus
Fig. 2. Effect of elevated CO2 concentrations on egg production rates (eggs
eggs.
female1 day1) of C. typicus.

the 385 ppm treatment on days 2 and 3 in week 30, but with very These results were confirmed statistically by the PERMANOVA
high variability (Fig. 5). analysis. Hatching success of C. typicus was not affected in any of
432 K. McConville et al. / Marine Pollution Bulletin 73 (2013) 428–434

50 Table 2
385 ppm Results of permutation-based analysis of variance for differences in feeding rate, egg
480 ppm
620 ppm production rate and hatching success of (a) Centropages typicus and (b) Temora
40
750 ppm longicornis (pp = ppm CO2, we = week, da(we) = day). Significant differences (p < 0.05)
9830 ppm
highlighted in bold.
egg production rate

Egg production Hatching success

30
Excluding CCS df f p df f p
(a) Centropages typicus
20 ppm 3 2.21 0.115 3 0.185 0.912
Week 3 3.064 0.079 3 4.346 0.002
Day 12 8.792 0.001 12 36.52 0.001
10 ppm  week 9 4.499 0.001 9 1.141 0.383
ppm  day 36 0.54 0.983 33 0.848 0.700
Res 123 115

0 Total 186 175

50 Including CCS df f p df p f
ppm 4 30.49 0.001 4 37.8 0.001
Week 3 3.49 0.071 3 4.12 0.005
40 Day 12 9.661 0.001 12 21.66 0.001
ppm  week 11 2.266 0.021 11 0.571 0.832
egg production rate

ppm  day 45 1.042 0.440 41 2.785 0.001

30 Res 147 135
Total 222 206

20 Excluding CCS df f p df f p
(b) Temora longicornis
ppm 3 0.732 0.559 3 0.501 0.700
10 Week 1 6.578 0.025 1 3.347 0.150
Day 6 21.170 0.001 6 1.537 0.190
ppm  week 1 2.409 0.122 1 0.819 0.400
0 ppm  day 12 0.849 0.608 11 1.397 0.220
Res 48 43
50
Total 71 65

Including CCS df f p df f p
40
ppm 4 0.61 0.643 4 0.908 0.471
egg production rate

Week 2 4.195 0.078 2 6.341 0.034

Day 9 22.18 0.001 9 1.439 0.200
30
ppm  week 1 2.484 0.119 1 0.45 0.521
ppm  day 15 0.858 0.605 14 1.836 0.061
Res 64 57
20
Total 95 87

10
5. Discussion

0 5.1. Effect of OA conditions on copepod reproduction

1 2 3 4
day
Fig. 4. Effect of elevated CO2 concentrations on egg production rates (eggs
female1 day1) of T. longicornis.
the OA treatments, but the reduction of hatched eggs in the CCS
treatment was significant (Table 2a). This was not the case for T.
longicornis, but the lack of significance may relate to the reduced
number of replicates for CCS conditions. The seasonal timing of
the experiment (week) had a significant effect on hatching success
when including the CCS incubations, while egg viability in the OA
treatments.was influenced by week in C. typicus only. The duration
of the exposures (day) also affected egg mortality in all treatments
in C. typicus. T. longicornis again showed no such effect and the var-
iability in the proportions of viable eggs was very high in weeks 29
and 30. A significant interaction occurred between pCO2 and day in
the CCS treatment of C. typicus. This interaction was not significant
for T. longicornis, but the p value was 0.06, and as such close to a
significant level.
In situ egg production differed between C. typicus and T. longi-
cornis, where C. typicus had much higher rates of on average 50
to >100 eggs female1 day1, while those of T. longicornis ranged
well below 50 eggs female1 day1. These dissimilarities reflect
the difference in body size and seasonal reproductive activity of
the two species at the outset of the experiments. Both species dis-
play marked seasonal cycles in egg production at mid-latitudes
(Ianora and Buttino 1990; Halsband and Hirche 2001; Halsband-
Lenk et al. 2001), with differing timing of maximal reproduction
based on their thermal preferences (Halsband-Lenk et al. 2002).
Our experiments fall into a period of maximal reproduction of C.
typicus, but low to moderate reproductive output of T. longicornis
(Halsband and Hirche 2001; Dam and Lopes 2003; Peters et al.
2007).
In the CO2 exposures egg production rates were mostly some-
what lower than in situ, and were significantly affected by the
length of the experiments (days). Egg production of both C. typicus
and T. longicornis decreased over time in several (but not all) exper-
iments, mirroring the result of Mayor et al. (2007). This may be ex-
plained by the limited capacity of females to store sperm and
 K. McConville et al. / Marine Pollution Bulletin 73 (2013) 428–434 433

1,0 both species hatched in week 31. Low hatching success may be
due to unsuitable food conditions encountered by copepod fe-
males. The availability and quality of maternal prey can affect
0,8
egg hatching success

the viability of copepod eggs (Arendt et al. 2005; Halsband-Lenk

et al. 2005; Jonasdottir et al. 2009).
0,6 The OA treatments chosen in experiments did not statistically
affect hatching success of C. typicus or T. longicornis. This is in
agreement with Kurihara and Ishimatsu (2008), who found no ef-
0,4 fect of 2380 ppm CO2 on the hatching success of Acartia tsuensis.
The duration of the incubations (day) had an effect on the hatching
385 ppm
0,2 480 ppm success of C. typicus but not T. longicornis, possibly due to a lower
620 ppm
750 ppm number of incubations for the latter species. Lower and more var-
9830 ppm iable hatching success later in the experiments may reflect the pro-
0,0 duction of unfertilized eggs, since females could not be re-
fertilized.
1,0 We did not investigate the effect of elevated pCO2 on fertiliza-
tion. However, negative effects on sperm motility, observed for
0,8 other marine organisms (Havenhand et al. 2008), are unlikely as
egg hatching success

fertilization is internal in calanoid copepods (Barthélémy, 1998;

Ianora et al. 2007).
0,6 This study did not allow for gradual acclimatization of adult
copepods to their respective elevated CO2 conditions. Copepods
0,4 were moved abruptly from control conditions to CO2 concentra-
tions predicted for the year 2100. It is likely that this sudden
change constitutes much greater physiological stress than the
0,2 gradual change of the entire ocean, thus supporting the view that
the reproduction of copepods is resilient to elevated CO2. Though
0,0 this abrupt change in conditions is likely to be more physiologi-
cally challenging than the gradual change expected, this is not an
1,0 accurate representation of how ocean acidification will develop,
so the inference with regard to OA is somewhat limited. The fitness
and viability of the juveniles produced in OA conditions was not
0,8
egg hatching success

measured, and it is possible that negative effects could manifest

over longer term incubations, or in different life stages.
0,6
5.2. Simulation of CCS conditions

0,4
0,2
0,0
1 2 3 4
day
Fig. 5. Effect of elevated CO2 concentrations on the proportion of viable T.
longicornis eggs.
fertilize consecutive clutches of eggs and/or by decreasing physio-
logical fitness over time, e.g. due to food limitation or female
senescence (Parrish and Wilson, 1978; Jonasdottir et al. 2009).
In isolation, no adverse effect of OA on egg production could be
detected in either species, similar to the results reported by Kuri-
hara et al. (2004) for Acartia congeners. The interactive effects be-
tween OA conditions and week, however, emphasize the
importance of seasonal timing for negative OA effects on reproduc-
tive traits. Mayor et al. (2012) found a similar, context-dependent
effect of OA on Calanus finmarchicus: temperature effects on hatch-
ing success depended on when the experiments were conducted.
The fitness of females in our experiments varied between weeks,
and copepods with lower starting fitness may be more susceptible
to elevated pCO2 than copepods in optimal physiological condition.
Hatching success in situ also differed somewhat between the
two copepods: it was >75% for C. typicus in weeks 26–28, but 6 75%
for T. longicornis in weeks 29 and 30. Around 90% of the eggs of
The results relating to the effect of CCS conditions on copepod
reproduction were similar to those recorded by Kurihara et al.
(2004). They found that both egg production rate and hatching suc-
cess of the copepods Acartia steueri and A. erythraea were reduced
through incubation in 10,000 ppm CO2. Egg viability was also im-
paired in Calanus finmarchicus at 8000 ppm (Mayor et al. 2007).
We found that incubation in 9830 ppm CO2 significantly reduced
egg production rate and hatching success of C. typicus. Neither trait
was affected in T. longicornis. The different result may reflect a spe-
cies-specific difference in response and higher resilience of T. lon-
gicornis to CCS conditions, but the difference in sampling effort
compared to C. typicus (number of incubated females: Centropages
n = 186, Temora n = 99) may alternatively suggest an insufficient
number of incubations to detect an effect. Kurihara et al. (2004) de-
tected a significant difference in the impact of lowered pH condi-
tions on egg production rate between Acartia species.
Our results also show an interactive effect between pCO2 and
day for hatching success when CCS concentrations are included
in the statistics (Table 2). The interaction was highly significant
for C. typicus (p 6 0.001) and just above the significance level for
T. longicornis (p = 0.061). This may be an indication that copepod
embryo mortality increases with longer exposure of their mothers
to deleterious concentrations of high pCO2.
Thus, copepod reproduction could be compromised by a CCS
leak. In areas with high CCS activity and regular or continuous
CO2 pollution, populations of calanoid copepods might be im-
pacted (see also Halsband and Kurihara, 2013). If this is the case
for relatively pH resistant organisms such as calanoid copepods,
the effect on less resistant zooplankton groups, in particular calci-
434 K. McConville et al. / Marine Pollution Bulletin 73 (2013) 428–434
fying organisms, may be severe. We also noted, however, that egg
viability of both species tended to recover towards the end of the
exposures. This may suggest an acclimation effect. To explore
how low pH acts as a directional selective force on copepod popu-
lations, together with synergistic or antagonistic effects of addi-
tional drivers of selection such as warming (Dam 2012), requires
experimentation over several generations of copepods. Differential
adaptation capacity of the various species and functional groups
will complicate the predictions of acidification impacts on plank-
ton communities.
6. Conclusions
The reproduction of healthy calanoid copepods seems unaf-
fected by gradually increasing pCO2 in an otherwise optimal envi-
ronment. During periods of adverse environmental conditions,
however, low fitness of female copepods and their offspring may
be further impaired by additional OA effects. Especially CCS leak-
ages with strong gradients in pCO2, may affect the fitness of adult
and embryonic copepods, and the effects – or their severity – may
be species-specific. On the other hand, those species with high tol-
erance to elevated pCO2 may have an advantage over more suscep-
tible competitors. Species with a large biogeographic range tend to
have greater physiological tolerance to changes in environmental
factors compared to those with a smaller range (Bozinovic et al.,
2011), giving generalists a potential advantage over specialists.
Whether this is also true for copepod pCO2 responses requires fur-
ther investigation. Differential species-specific and time-depen-
dent effects, as shown by our study, may consequently impact on
the composition and structure of future zooplankton communities
in a high CO2 ocean.
Acknowledgments
We thank the crew of the RV Quest for collecting plankton sam-
ples, Rachel Harmer and Andrea McEvoy for their help in picking
out copepods and three anonymous reviewers for helpful com-
ments on an earlier version of this manuscript. This work forms
part of the Natural Environment Research Council (NERC), Plym-
outh Marine Laboratory core Strategic Research Programme under
Oceans 2025 and NERC grant no. J1083106 in the UK Ocean Acid-
ification Programme. It was also supported by the Norwegian Fram
Centre Flagship for Ocean Acidification.
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