Documentos de Académico
Documentos de Profesional
Documentos de Cultura
DOI 10.1007/s00227-016-2905-z
ORIGINAL PAPER
Received: 21 September 2015 / Accepted: 26 April 2016 / Published online: 9 May 2016
Springer-Verlag Berlin Heidelberg 2016
13
131
Page 2 of 13 Mar Biol (2016) 163:131
(Tamayo etal. 2011, 2013, 2014, 2015), or 2) the capacity the specific physiological mechanisms involved in size
to save higher amounts of energy due to reduced metabolic differentiation and to compare these mechanisms among
rates (combination 2). the different conditions applied to the selection of growth
A high degree of multi-locus heterozygosity is an groups.
endogenous factor widely correlated with enhanced growth
rates among bivalves. Variable levels of heterozygosity
appear to underlie the physiological traits responsible for Materials andmethods
fast growth in combination 1 (Holey and Foltz 1987; Toro
and Vergara 1998; Pace etal. 2006; Meyer and Manahan Collection, maintenance andselection offast (F)
2010) and in combination 2 (Garton etal. 1984; Hawkins andslow (S)growing mussels
etal. 1986; Hawkins 1995; Bayne and Hawkins 1997).
Although heterozygositygrowth associations in bivalves In both Expt. 1 and Expt. 2, juvenile (<16mm shell length,
are widely accepted (see reviews: David 1998; Launey SL) mussels (Mytilus galloprovincialis) were collected
and Hedgecock 2001; Meyer and Manahan 2010), cor- from the rocky intertidal in the Plentzia estuary in Biscay,
relations reported in the literature seem intrinsically weak Spain (43 24 42,462 N 02 56 43,659 W).
(reviewed by David 1998) and frequently explain 2030%
of observed variance, at the most (Garton 1984; Koehn and Expt. 1: restrictive feeding conditions
Gaffney 1984; Diehl and Koehn 1985; Gentili and Beau-
mont 1988; Koehn etal. 1988; Alvarez etal. 1989; David Two hundred freshly collected mussels (November 16,
etal. 1995). Increased multi-locus heterozygosity enhances 2007), 716mm SL umbo to margin, were measured to the
the energy budget of fast growers through improved effi- nearest 0.01mm with a calliper and distributed in nine 1
ciency of protein deposition by reducing protein turnover mm SL groups (i.e. 7.007.99mm, 8.008.99mm and so
rates. This mechanism leads to more efficient standard and on). These groups were maintained separately for 5months
routine metabolic rates (Hawkins etal. 1986, 1989; Bayne with a low ration of phytoplankton (Isochrysis galbana,
and Hawkins 1997). T-ISO, given at a packed volume concentration of parti-
Interpreting those combinations would benefit by relat- cles of 0.5mm3 L1; equivalent to 0.22mg POM L1)
ing the environment to the physiological characteristics in a common feeding tank inside a recirculating sea water
resulting in faster growth. In this respect, food conditioning system regulated at 14C (environmental temperature at
seems to affect the association between genetic and physi- time of collection) and 34.0 salinity. In order to simulate
ological traits (Bayne and Hawkins 1997), lending weight the discontinuous feeding imposed by the tidal regime at
to the argument that benefits of a reduced protein turnover the collection site (about 50% emersion), mussels were fed
are likely dependent on available food levels. As stated by phytoplankton concentrations stated above for 12h day1.
Bayne and Hawkins (1997), genetic factors promoting dif- During the maintenance period, SL of individuals in each
ferential protein turnover requirements become relevant size group was measured on nine occasions.
when food is abundant (when energy incorporation domi- At the end of the maintenance period, F and S mussels
nates energy balance), whereas heterozygosity and growth were selected by choosing the smallest and largest individ-
rate are uncorrelated at low food rations (when mainte- uals (~P25 and P75 percentiles) in each size group. Individ-
nance requirements are the determinant). ual growth rates were estimated as a daily increment (%)
The aim of the present study was to test the hypothesis using the following formula:
that the physiological components acting on size differ-
100 ([final SLinitial SL]/initial SL)/t (day)
entiation of fast- versus slow-growing individuals differ
as a function of nutritional conditions during the selective where final SL is the average SL after 5months of mainte-
phase. For this purpose, juvenile Mytilus galloprovincia- nance, and initial SL is the average SL of their size group at
lis collected from a natural population were maintained in the beginning. F and S groups used in physiological experi-
the laboratory until size differentiation occurred under two ments consisted of 30 fast- and 30 slow-growing mus-
different experimental conditions: low and intermittent sels selected following this methodology. These individu-
phytoplankton supply involving restrictive feeding con- als were between 12.2519.29mm SL in the F group and
ditions (Experiment 1, Expt. 1), and high and continuous 10.7016.17mm SL in the S group.
phytoplankton ration involving optimal feeding conditions
(Experiment 2, Expt. 2). In both experiments, individual Expt. 2: optimal feeding conditions
with lowest and highest growth rates were selected and the
physiological components of their energy balances deter- In this experiment, 200 mussels (March 19, 2009),
mined at different food rations. The goal was to identify 1015mm SL, were tagged for individual growth rate
13
Mar Biol (2016) 163:131 Page 3 of 13 131
Table1Characteristics of experimental diets used in physiological organic matter (mg L1), OC organic content (decimal), Vol. packed
determinations in Expt. 1 and 2: TPM total particulate matter (mg volume (mm3L1), n number of samples
L1), PIM particulate inorganic matter (mg L1), POM particulate
1
Restrictive feeding condition Low (RL) 0.730.07 0.060.04 0.670.04 0.920.05 1.590.11 5
High (RH) 1.320.08 0.130.05 1.190.05 0.90.03 3.20.13 8
2
Optimal feeding condition Low (OL) 0.590.18 0.140.04 0.450.18 0.750.07 0.790.17 11
High (OH) 1.690.25 0.520.09 1.170.18 0.70.03 3.070.32 9
13
131
Page 4 of 13 Mar Biol (2016) 163:131
These flow rates were adjusted to ensure reliability of CR Newell 1983, for discussion on using the concept in physio-
measurements while precluding water re-circulation by the logical energetics of bivalves). Regarding our experimental
mussels before leaving the system, which would cause CR conditions for RMR and SMR, we have proposed (Tamayo
underestimation. The particulate suspension in the feeding etal. 2013) the more specific term MSFG (metabolic scope
tank was maintained at the target concentration for each for feeding and growth) to denote the difference between
ration by addition of the appropriate volume of particles the two rates.
from concentrated stocks of the corresponding diet, while Rates of oxygen consumption were converted into
concentration (mm3 L1) was frequently checked with a metabolic rates using an oxycaloric coefficient of 20.08J
Coulter Multisizer 3. A control chamber (without mussels) mLO1 2 (Gnaiger 1983).
served to correct particle sedimentation.
Mussels were given a 3-day acclimation period in these Energy balance
chambers prior to measurements. During the fourth day,
CR measurements with each chamber were replicated 56 Resulting energy balance was computed as the scope
times and faeces (for AE determinations) were collected for growth (SFG: Jh1), which represents the difference
on three occasions. Sample values represent the average of between absorption rate (AR: Jh1) and routine metabolic
these replicates. rate (RMR: Jh1).
In Expt. 1 (RL and RH conditions), 5 samples (cham-
bers) were used (n =5) per condition and growth group, Size standardization
each composed of 2 or 3 individuals of similar size with
similar growth rates. Physiological rates in this case were After physiological determinations, mussels were dissected
referred to the mean individual weight of the group. In out of the shells for flesh dry weight (DW) determination.
Expt. 2 (OL and OH conditions), 10 individuals (n =10) Clearance and metabolic rates were standardized to a com-
were used per condition and growth group. In both experi- mon size of 50mg DW, which represents the average dry
ments, different F and S individuals were used to perform weight of F and S mussels in Expt. 1 and 2,by using the
measurements at high and low rations. following expression:
Clearance rates (CR: L h1) were calculated according
to Crisp (1971) with the following formula: Yst = (Wstd /We )b Ye
where Yst and Ye represent the standard and experimentally
CR = (F/n) ((Ci Co)/Ci)
recorded rates, respectively, We the weight of the experi-
where F is the flow rate (L h1), n is the number of individ- mental mussels, Wstd the standard weight and b the power
uals per sample, and Ci and Co are the particle concentra- value that scales physiological rates to body weight in
tions in the outflows of control and experimental chambers, this species, 0.583 for CR (Bayne and Hawkins 1997) and
respectively. 0.724 for metabolism (Bayne etal. 1973).
Absorption efficiencies (AE: decimal units) were deter-
mined following Conover (1966), by comparing the organic Statistical procedures
content of food samples taken from the feeding tanks and
faeces collected from the bottom of the chambers. Significant effects with respect to growth category (F vs.
S) and food ration on standardized physiological param-
Metabolic expenditure eters were tested using a two-factor analysis of variance
(ANOVA). Significant differences between values within
Once CR and AE measurements were completed, mussels experiments were checked by means of t test comparisons.
were transferred to respirometers (150-ml capacity) filled A three-factor analysis of variance (ANOVA) was per-
with filtered sea water and sealed with LDO oxygen probes formed to test significant differences among growth catego-
(cQ40d), and oxygen consumption was computed from ries (F vs. S), food rations and experiments.
the decrease in oxygen concentration over time (12h). Only in the case of RL, RMR and SMR measurements
Controls (chambers without mussels) were used to check were carried out on different individuals, and thus, MSFG
the stability of oxygen concentration during the measure- was computed without an associated variance. With the
ment period. These measurements corresponded to the rou- aim of unifying the analysis of MSFG variation, signifi-
tine metabolic rate (RMR: Jh1). For standard metabolic cant differences in metabolic rates promoted by activity
rate (SMR: Jh1), mussels were starved for 48h before level (routine vs. standard) and growth category were
measuring their oxygen consumption again. The difference tested with a repeated measures two-factor ANOVA for
between resting and active levels of metabolism is com- RH, OL and OH conditions, and a standard two-factor
monly referred to as the scope for activity (see Bayne and ANOVA for RL.
13
Mar Biol (2016) 163:131 Page 5 of 13 131
13
131
Page 6 of 13 Mar Biol (2016) 163:131
Table2Expt. 1. Physiological variables in fast (F)- and slow (S)-growing mussel juveniles selected in a restrictive food regime, tested at low
and high rations
Physiological parameter Growth category Low ration High ration Summary of two-factor ANOVA
Growth category Ration Interaction
OIR (mgh1)
F a 0.2880.022 a b 0.2650.034 DF=1 DF=1 DF=1
S b 0.2240.023 a b 0.2500.014 F=9.21 F=0.013 F=3.57
p=0.009 p=0.909 p=0.080
AE (decimal)
F b 0.3690.052 a 0.5470.052 DF=1 DF=1 DF=1
S b 0.3980.025 a 0.5320.025 F=0.082 F=47.51 F=0.932
p=0.779 p=0.000 p=0.351
AR (Jh1)
F bc 1.9790.235 a 2.6980.361 DF=1 DF=1 DF=1
S c 1.6550.087 a b 2.4790.025 F=4.63 F=37.44 F=0.173
p=0.049 p=0.000 p=0.684
RMR (Jh1)
F a 1.0760.131 a 1.1510.205 DF=1 DF=1 DF=1
S a 0.9490.118 a 1.1470.366 F=0.34 F=1.50 F=0.30
p=0.566 p=0.241 p=0.590
SFG (Jh1)
F ab 0.9030.228 a 1.5480.402 DF=1 DF=1 DF=1
S b 0.7060.180 a b 1.3330.389 F=1.56 F=14.88 F=0.003
p=0.232 p=0.002 p=0.957
SMR (Jh1)
F b 0.6350.116 a b 0.7730.180 DF=1 DF=1 DF=1
S ab 0.7670.085 a 0.9730.150 F=5.14 F=5.54 F=0.22
p=0.040 p=0.034 p=0.649
MSFG (Jh1)
F 0.440 a 0.3780.339
S 0.182 a 0.1740.466
OIR organic ingestion rate, AE absorption efficiency, AR absorption rate, RMR routine metabolic rate, SFG scope for growth, SMR standard met-
abolic rate, MSFG metabolic scope for feeding and growth. Physiological rates were standardized to an equivalent 50mg dry soft-body weight.
Mean values are presented95% C.I. (n=5 and 4 for F and S groups, respectively) together with a summary of two-factor ANOVA testing
significant effects of growth category and experimental food ration. Different letters beside each mean value indicate significant differences
among FL, SL, FH and SH (t test, p<0.05)
was calculated as the product of OIR and AE, it was sig- experiments separately. Namely, F mussels had higher food
nificantly affected by both growth category and food ration processing capabilities that ultimately lead to higher absorp-
(ANOVA, Table3). Thus, overall, F mussels had a higher tion rates (AR), and ration increments enhanced energy
AR than S juveniles even though their ration specific values acquisition. Interestingly, the factor termed experiment (i.e.
did not differ significantly (10 and 27% in OL and OH, Expt. 1 vs. 2) interacted significantly with food ration, likely
respectively). Moreover, both groups increased their AR in due to the wider food range achieved in Expt. 2.
a similar fashion with increasing food concentration (2.2
and 1.9 times for F and S, respectively). Metabolic expenditure
A summary of a 3-factor ANOVA to test for significant
effects promoted by growth category (F vs. S), food ration Mean values of metabolic rates under routine and stand-
(L vs. H) and experiment (Expt. 1 vs. Expt. 2), upon physi- ard conditions (RMR and SMR) are given in Table2 and 3
ological parameters of the energy balance, can be seen in for mussels conditioned to restrictive and optimal rations,
Table 4. This analysis offered, in general terms, an inter- respectively. Results of both experiments are compared
pretation of the results similar to those reported for the two (Fig. 3), with metabolic scope for feeding and growth
13
Mar Biol (2016) 163:131 Page 7 of 13 131
0.3
b c tal rations (RL, RH, OL and OH; Table5). Tested factors
0.2
were: 1) growth category (F vs. S) and 2) the level of meta-
c bolic activity (SMR vs. RMR). The latter factor was used
0.1 to test whether metabolic differences between starved and
0
fed conditions of mussels (in fact, MSFG) were statistically
0.4 0.7 1 1.3 significant. Therefore, the interaction term in present analy-
sis accounts for significant differences in MSFG between F
0.5 b and S mussels.
0.4 a According to the ANOVA, feeding and growth activities
ab promoted significant metabolic increments in comparison
CR (L h-1)
0.3
with starved conditions in all cases except for high ration
a
0.2 in Expt. 1 (restrictive food conditioning, RH), where it was
b not statistically significant (p=0.095; Table5). However,
0.1
the interaction between metabolic level and growth cat-
0 egory was at the threshold of significance (p =0.05) for
0.4 0.7 1 1.3
this RH experimental condition, suggesting that metabolic
POM (mg L-1)
behaviour during starvation differed between F and S mus-
sels. In fact, a t test to compare metabolic levels within
Fig.2Size standardized clearance rate (CR: L h1) as a function of growth categories in this particular condition revealed sig-
particulate organic matter (POM: mg L1) for F (circles) and S (tri-
angles) Mytilus galloprovincialis selected under a restrictive food nificant differences between routine (RMR) and standard
regimes (Expt. 1, n=5 and 4 for F and S groups, respectively); and (SMR) metabolic rates for F mussels (t test, t8 =2.781;
b optimal food regimes (Expt. 2 n =8 and 10 for F and S groups, p =0.024) and lack of significance for S mussels (t test,
respectively). Values are means95% C.I. CR was standardized to t6=-0.578; p=0.584). Overall, analysis of factor interac-
an equivalent 50mg dry soft-body weight. Different letters indicate
significant differences (t test) tions (Table5) reveals some features about the metabolic
behaviour of F and S mussels subjected to different food
conditioning regimes. As a general rule, although the mag-
(MSFG) values reported as a fraction of RMR. In Expt. 1, nitude of MSFG differs between F and S mussels, the sign
there were no significant effects on routine rates of growth of this difference is strongly dependent on food condition-
category or ration. However, standard rates a) were higher ing: i.e. MSFG would be greater in F than in S mussels
in S when compared to F mussels and b) increased with if maintained under restrictive food conditions and vice
the higher food ration in both types of mussels (Table2). versa for F and S mussels conditioned to an optimal ration
In Expt. 2, no significant effects were recorded on stand- (Fig.3). Strict interpretation of the ANOVA in Table5 indi-
ard rates, while routine rates only differed significantly cates, however, that this was significant only for metabolic
between low and high rations (Table3). A 3-factor measurements performed in mussels at low rations.
ANOVA (Table4) confirmed that food ration was the only
factor exerting a significant effect on RMR. Regarding Scope forgrowth
the standard rate (SMR), both ration and the interaction
between growth category and experiment exerted signifi- SFG of mussels conditioned to restrictive food conditions
cant effects. These results account for the fact that, over- was positively influenced by food concentration, effects
all (1) food ration tends to increase SMR, and (2) while of ration being significant (Table2). Although consist-
F individuals had similar SMRs in both experiments, S ently higher in F mussels, SFG differences between growth
mussels had higher rates than F mussels in Expt. 1 and groups were not significant (Table2); the benefits to fast
lower in Expt. 2. growers of achieving significantly higher absorption rates
With respect to metabolic scope for feeding and growth (especially at low POM) were counterbalanced by higher
(MSFG), no differences were found between F and S mus- metabolic expenditures.
sels in Expt. 1 (RH rations) (Table2). However, in Expt. 2 Under optimal food conditioning, SFG was significantly
both growth category and ration were statistically signifi- affected by both growth category and food ration (Table3).
cant. While higher rations caused a significant increase in In addition to a positive influence of POM, large differ-
MSFG for S mussels, similar values were recorded for F ences between F and S mussels were recorded, reflecting
individuals (Table3). Further statistical analysis of MSFG the combined effects of a greater capacity of F mussels to
13
131
Page 8 of 13 Mar Biol (2016) 163:131
Table3Expt. 2. Physiological variables in fast (F)- and slow (S)-growing mussel juveniles selected in an optimal food regime, tested at low and
high rations
Physiological parameter Growth category Low ration High ration Summary of two-factor ANOVA
Growth category Ration Interaction
OIR (mgh1)
F c 0.1510.016 a 0.3070.040 DF=1 DF=1 DF=1
S c 0.1440.023 b 0.2330.048 F=4.99 F=44.76 F=3.35
p=0.032 p=0.000 p=0.077
AE (decimal)
F bc 0.5420.025 ab 0.5940.019 DF=1 DF=1 DF=1
S c 0.5200.058 a 0.6180.015 F=0.007 F=16.23 F=1.52
p=0.935 p=0.000 p=0.226
AR (Jh1)
F b 1.5290.156 a 3.4070.454 DF=1 DF=1 DF=1
S b 1.4090.263 a 2.6850.542 F=4.25 F=59.62 F=2.17
p=0.047 p=0.000 p=0.150
RMR (Jh1)
F b 0.8370.139 a 1.3270.243 DF=1 DF=1 DF=1
S ab 1.0440.205 a 1.4780.259 F=2.45 F=16.36 F=0.062
p=0.127 p=0.000 p=0.805
SFG (Jh1)
F bc 0.6920.161 a 2.0800.401 DF=1 DF=1 DF=1
S c 0.3640.265 b 1.2070.614 F=7.76 F=26.79 F=1.60
p=0.009 p=0.000 p=0.215
SMR (Jh1)
F a 0.6870.117 a 0.8210.159 DF=1 DF=1 DF=1
S a 0.5400.126 a 0.6920.256 F=2.177 F=2.35 F=0.10
p=0.150 p=1.135 p=0.992
MSFG (Jh1)
F b 0.1500.147 ab 0.5070.328 DF=1 DF=1 DF=1
S ab 0.5040.181 a 0.7850.231 F=7.29 F=7.43 F=0.103
p=0.011 p=0.010 p=0.750
OIR organic ingestion rate, AE absorption efficiency, AR absorption rate, RMR routine metabolic rate, SFG scope for growth, SMR standard met-
abolic rate, MSFG metabolic scope for feeding and growth. Physiological rates were standardized to an equivalent 50mg dry soft-body weight.
Mean values are presented95% C.I. (n=8 and 10 for F and S groups, respectively) together with a summary of two-factor ANOVA testing
significant effects of growth category and experimental food ration. Different letters beside each mean value indicate significant differences
among FL, SL, FH and SH (t test, p<0.05)
absorb energy and reduce metabolic expenditure by hav- differences in growth and, consequently, allowing us to
ing significantly lower costs associated with growth repre- form groups with very different growth rates (approxi-
sented by MSFG (Table5; Fig.3). mately 3 times higher for fast growers: 1.240.24 vs.
0.44 0.19% increment in SL day1 for F and S mus-
sels, respectively). In contrast, the low food conditions in
Discussion Expt. 1 promoted low growth rates reducing the scope for
size differentiation. Nevertheless, after 5months, large size
The present study used significantly size-differentiated differences were attained and the two groups selected for
mussels maintained under food abundance or food depri- the highest (F) and the lowest (S) growth rates achieved
vation for sustained periods in the laboratory. Maintenance 0.35 0.17% and 0.050.03 increment in SL day1,
of mussels under conditions of high phytoplankton supply respectively.
(Expt. 2) intensified the overall growth potential of organ- Comparison of physiological parameters involved in the
isms, thus forcing the emergence of large interindividual energy balance between the different growth groups in the
13
Mar Biol (2016) 163:131 Page 9 of 13 131
13
131
Page 10 of 13 Mar Biol (2016) 163:131
13
Mar Biol (2016) 163:131 Page 11 of 13 131
Table5Summary of two-factor ANOVA testing for significant heterozygosity and growth rate at low rations suggests that
effects of a) level of activity (routine vs. standard) and, b) growth cat- heterozygosity level does not modulate standard metabolic
egory (F vs. S) on metabolic rates in mussels from the four combina-
tions of experimental conditions (RL, selected under restrictive food expenditure. Therefore, it is very likely that interindividual
conditions and fed low rations; RH, selected under restrictive food differences in SMR derive from causes other than differ-
conditions and fed high rations; OL, selected under optimal food con- ences in protein deposition efficiencies. In this respect,
ditions and fed low rations; OH, selected under optimal food condi- Pernet etal. (2008) reported that intra-specific differences
tions and fed high rations)
in SMR are related to the unsaturation index of membrane
Experimental condition Source of variation DF F p fatty acids in Crassostrea virginica. These authors reported
RL Level of activity 1 26.550 0.000
that both SMR and membrane unsaturation were lower
in fast-growing hatchery oysters compared with slow-
Growth category 1 0.001 0.971
growing wild oysters. The higher unsaturation index in
Interaction 1 4.580 0.050
the latter corresponded mainly with higher 22:6n-3 levels
RH Level of activity 1 3.710 0.095
whose importance in metabolic rate regulation of animals
Growth category 1 1.240 0.303
had already been observed (reviewed by Hulbert and Else
Interaction 1 0.506 0.500
2005). Wild oysters had higher SMRs even though they
OL Level of activity 1 28.030 0.000
were much more heterozygous than fast-growing hatchery
Growth category 1 0.101 0.755
oysters with lower SMRs and higher CRs.
Interaction 1 8.220 0.011
Very likely the possession of a reduced maintenance
OH Level of activity 1 42.130 0.000
metabolic output improves the probability of surviving
Growth category 1 0.006 0.940
under stressful conditions. Diehl etal. (1986) grew juve-
Interaction 1 1.960 0.181
nile Mytilus edulis in a tidal salt marsh for 72 days and
For RL conditions, independent groups of mussels were used for met- then starved them in the laboratory for 2months. Recorded
abolic determinations; thus, a standard ANOVA was performed. For interindividual variability in rates of oxygen consumption
the rest of experimental conditions (RH, OL and OH), we performed suggested that reduced rates of metabolism were associated
an ANOVA of repeated measures, as the same groups of mussels
were used in routine and standard measurements
with increased resistance to weight loss during starvation
rather than with the capacity for weight gain during feed-
ing. An elegant experiment by LeBlanc etal. (2008) is even
under restrictive food levels (Hawkins 1995; Hawkins and more relevant. Juveniles of Mytilus edulis were submitted
Day 1996). to stressful conditions consisting in 11h of air exposure
Our study shows that food conditions during the size dif- followed by 6h of exposure to 32 C water, which caused
ferentiation phase might potentially alter the set of physi- high mussel mortality. Survivors were used to determine
ological variables underlying differential growth potential. SFG and multilocus heterozygosity and compared with
This fact, that feeding conditions can affect the correla- control mussels. Higher heterozygosity in survivors than
tions between endogenous (genetic) factors and physiologi- controls indicated that heterozygotes were somehow more
cal rates, was previously suggested by Bayne and Hawk- resistant to stress. Measurements of SFG resulted in non-
ins (1997). These authors analysed possible correlations significant differences between treated and control mussels,
between growth rate and multilocus heterozygosity in but, meaningfully, there were significant differences in both
small mussels that were sequentially held at increasing RMR and SMR which were, respectively, 40 and 30%
food levels. Increasing heterozygosity levels led to higher lower for survivors than for control mussels.
efficiencies of protein deposition and, as a consequence, A pertinent conclusion is that size differentiation of
lower costs of growth, thus allowing more heterozygous juvenile mussels in the natural environment results from a
individuals to achieve higher growth rates. These series of balance between opposing forces. These include a) periods
correlations served to highlight the functional link between of good trophic conditions characterized by high POM con-
protein turnover rate, metabolic efficiency and genetic fac- centrations and long immersion times that would maximize
tors. However, Bayne and Hawkins (1997) also showed growth rates of individuals well equipped to acquire food
that significant correlations between heterozygosity and and process it at low costs, in spite of their relatively high
growth rate were only found at high food rations, while resting metabolic rates, and b) periods of restricted trophic
at lower rations, corresponding to maintenance conditions conditions of both low POM, low organic content of food
(low SFG), no significant correlations were found. It fol- and/or prolonged emersion times that would have a more
lows that genetic factors promoting differential protein adverse impact on those individuals unable to down-reg-
turnover requirements become relevant under conditions of ulate their SMRs. This dual impact of feeding conditions
food abundance (i.e. when energy incorporation dominates upon the physiological profiles of individuals that are likely
the energy balance), whereas lack of correlation between to grow faster might help understanding contradictory
13
131
Page 12 of 13 Mar Biol (2016) 163:131
conclusions in previous bivalve studies regarding the preva- Gentili MR, Beaumont AR (1988) Environmental stress, heterozygo-
lence of energy input or output as the processes responsible sity, and growth rate in Mytilus edulis L. J Exp Mar Biol Ecol
120:145153
for size differentiation. Gnaiger E (1983) Heat dissipation and energetic efficiency in animal
anoxibiosis: economy contra power. J Exp Zool 228:471490
AcknowledgmentsThis study was funded through the project Hawkins AJS (1995) Effects of temperature change on ectotherm
AGL2009-09981 of the Spanish Ministry of Science and Innovation. metabolism and evolution: metabolic and physiological interrela-
D.T. was funded by a FPI grant from the Basque Government. Authors tions underlying the superiority of multi-locus heterozygotes in
are indebted to two referees for valuable comments and sugges- heterogeneous environments. J Therm Biol 2:2333
tions.Finally, Tamayo D. wants to thank, in particular, the invaluable Hawkins AJS, Day AJ (1996) The metabolic basis of genetic differ-
support received from Muguruza C. over the years. Will you marry me? ences in growth efficiency among marine animals. J Exp Mar
Biol Ecol 203:93115
Hawkins AJS, Bayne BL, Day AJ (1986) Protein turnover, physi-
References ological energetics and heterozygosity in the blue mussel Myti-
lus edulis: the basis of variable age-specific growth. Proc R Soc
Alvarez G, Zapata C, Amaro R, Guerra A (1989) Multilocus heterozy- Lond 229B:161176
gosity at protein loci and fitness in the European oyster, Ostrea Hawkins AJS, Widdows J, Bayne BL (1989) The relevance of whole-
edulis L. Heredity 63:359372 body protein metabolism to measured costs of maintenance and
Bayne BL (1999) Physiological components of growth differences growth in Mytilus edulis. Physiol Zool 62:745763
between individual oysters (Crassostrea gigas) and a comparison Holey ME, Foltz DW (1987) Effect of multiple-locus heterozygosity
with Saccostrea comercialis. Physiol Biochem Zool 72:705713 and salinity on clearance rate in a brackish water clam, Rangia
Bayne BL (2000) Relations between variable rates of growth, meta- cuneata (sowerby). J Exp Mar Biol Ecol 111:121131
bolic costs and growth efficiencies in individual Sydney rock oys- Hulbert AJ, Else PL (2005) Membranes and the setting of energy
ters (Saccostrea comercialis). J Exp Mar Biol Ecol 251:185203 demand. J Exp Biol 208:15931599
Bayne BL, Hawkins AJS (1997) Protein metabolism, the costs of Koehn RK, Gaffney PM (1984) Genetic heterozygosity and growth
growth, and genomic heterozygosity: experiments with the mus- rate in Mytilus edulis. Mar Biol 82:17
sel Mytilus galloprovincialis Lmk. Phys Zool 70:391402 Koehn RK, Diehl WJ, Scott TM (1988) The differential contribution
Bayne BL, Newell RC (1983) Physiological energetics of marine by individual enzymes of glycolysis and protein catabolism to
moluscs. In: Saleudden ASM, Wilbur KM (eds) The Mollusca. the relationship between heterozygosity and growth rate in the
Academic Press, New York 4(1): 407515 coot clam, Mulinia lateralis. Genetics 118:121130
Bayne BL, Thompson RJ, Widdows J (1973) Some effects of tem- Launey S, Hedgecock D (2001) High genetic load in the Pacific oys-
perature and food on the rate of oxygen consumption by Mytilus ter Crassostrea gigas. Genetics 159:255265
edulis L. In: Wieser W (ed) Effects of temperature on ectother- LeBlanc N, Tremblay R, Davidson J, Landry T, McNiven M (2008) The
mic organisms. Springer, Berlin, pp 181193 effect of selection treatments on Mytilus edulis, modifications of
Bayne BL, Hawkins AJS, Navarro E, Iglesias JIP (1989) Effects of genetic and physiological characteristics. Mar Biol 153:11411152
seston concentration on feeding, digestion and growth in the Meyer E, Manahan DT (2010) Gene expression profiling of geneti-
mussel Mytilus edulis. Mar Ecol Prog Ser 55:4754 cally determined growth variation in bivalve larvae (Crassostrea
Bayne BL, Hedgecock D, McGoldrick D, Rees R (1999a) Feeding gigas). J Exp Biol 213:749758
behaviour and metabolic efficiency contribute to growth hetero- Pace DA, Marsh AG, Leong Patrick K, Green AJ, Hedgecock D,
sis in Pacific oysters [Crassostrea gigas (Thunberg)]. J Exp Mar Manahan DT (2006) Physiological bases of genetically deter-
Biol Ecol 233:115130 mined variation in growth of marine invertebrate larvae: a study
Bayne BL, Svensson S, Nell JA (1999b) The physiological basis for of growth heterosis in the bivalve Crassostrea gigas. J Exp Mar
faster growth in the Sydney rock oyster, Saccostrea commercia- Biol Ecol 335:188209
lis. Biol Bull 197:377387 Parry GD (1983) The influence of the costs of growth on ectotherm
Conover RJ (1966) Assimilation of organic matter by zooplankton. metabolism. J Theor Biol 101:453477
Limnol Oceanogr 11:338354 Pernet F, Tremblay R, Redjah I, Svigny JM, Gionet C (2008) Physi-
Crisp DJ (1971) Energy flow measurements. In: Holme NA, Mc ological and biochemical traits correlate with differences in
Inture AD (eds) Methods for the study of marine benthos, (IBP growth rate and temperature adaptation among groups of the
no 16). Blackwell, Oxford, pp 197323 eastern oyster Crassostrea virginica. J Exp Biol 211:969977
David P (1998) Heterozygosityfitness correlations: new perspec- Tamayo D, Ibarrola I, Urrutia MB, Navarro E (2011) The physiologi-
tives on old problems. Heredity 80:531537 cal basis for inter-individual growth variability in the spat of
David P, Delay B, Berthou P, Jarne P (1995) Alternative models for clams (Ruditapes philippinarum). Aquaculture 321:113120
allozyme-associated heterosis in the marine bivalve Spisula ova- Tamayo D, Ibarrola I, Navarro E (2013) Thermal dependence of
lis. Genetics 139:17191726 clearance and metabolic rates in slow- and fast-growing spats
Diehl WJ, Koehn RK (1985) Multiple-locus heterozygosity, mortality of manila clam Ruditapes philippinarum. J Comp Physiol B
and growth in a cohort of Mytilus edulis. Mar Biol 88:256271 183:893904
Diehl WJ, Gaffney PM, Koehn RK (1986) Physiological and genetic Tamayo D, Ibarrola I, Urrutxurtu I, Navarro E (2014) Physiological
aspects of growth in the mussel Mytilus edulis. I. Oxygen con- basis of extreme growth rate differences in the spat of oyster
sumption, growth and weight loss. Physiol Zool 59:201211 (Crassostrea gigas). Mar Biol 161:16271637
Garton DW (1984) Relationship between multiple locus heterozygo- Tamayo D, Ibarrola I, Navarro E (2015) The effects of food condition-
sity and physiological energetics of growth in the estuarine gas- ing on feeding and growth responses to variable rations in fast
tropod Thais haemastoma. Physiol Zool 57:530543 and slow growing spat of the Manila clam (Ruditapes philippi-
Garton DW, Koehn RK, Scott TM (1984) Multiple-locus heterozy- narum). J Exp Mar Biol Ecol 471:92103
gosity and the physiological energetics of growth in the coot Thompson RJ, Bayne BL (1974) Some relationships between growth,
clam, Munilia lateralis, from a natural population. Genetics metabolism and food in the mussel Mytilus edulis. Mar Biol
108:445455 27:317326
13
Mar Biol (2016) 163:131 Page 13 of 13 131
Toro JE, Vergara AM (1998) Growth and heterozygosity in a Whyte J (1987) Biochemical composition and energy content of six
12-month-old cohort of Ostrea chilensis obtained by mass species of phytoplankton used in mariculture of bivalves. Aqua-
spawning in laboratory. P S Z N Mar Ecol 19:311323 culture 60:231241
13