Aquaculture 304 (2010) 34–41
Contents lists available at ScienceDirect
Aquaculture
j o u r n a l h o m e p a g e : w w w. e l s ev i e r. c o m / l o c a t e / a q u a - o n l i n e
Nutritional vulnerability and compensatory growth in early juveniles of the “red
claw” crayfish Cherax quadricarinatus
Liane Stumpf, Natalia S. Calvo, Silvia Pietrokovsky, Laura S. López Greco ⁎
Biology of Reproduction and Growth in Crustaceans. Dept. of Biodiversity and Experimental Biology, FCEyN, University of Buenos Aires, Cdad. Univ. C1428EHA, Buenos Aires, Argentina
a r t i c l e
i n f o
Article history:
Received 18 January 2010
Received in revised form 26 February 2010
Accepted 9 March 2010
Keywords:
Cherax quadricarinatus
Juveniles
Point-of-reserve-saturation
Point-of-no-return
Intermittent feeding
Compensatory growth
a b s t r a c t
The objective of this study was to investigate the nutritional vulnerability of stage III juveniles of the
freshwater crayfish Cherax quadricarinatus. The point-of-reserve-saturation (PRS50), the point-of-no-return
(PNR50) and the compensatory growth capacity in intermittently fed individuals were determined. The
juveniles were subjected to different feeding protocols for the PRS–PNR experiment and the intermittent
feeding experiment. Fifty percent of the juveniles were able to successfully molt to the following stage
(PRS50) after 3.53 feeding days and failed to molt to the following stage (PNR50) after 4.28 starvation days. In
the intermittent feeding experiment, stage III juveniles achieved similar growth and survival compared to
those kept under the continuous feeding regimen. Full compensatory growth occurred during the restriction
period in the initially fed group and during the refeeding period in the initially starved group. This is the first
study on PRS, PNR and compensatory growth in a subtropical freshwater crayfish. Moreover, it shows that
stage III juveniles are unable to molt to the following stage without feeding, requiring at least two feeding
days to molt and survive. Feeding seems to be necessary at the beginning of stage III, when energy reserves
are probably accumulated. On the other hand, the capacity to fully compensate may lead to a reduction in the
amount of food supplied, thereby diminishing the costs of production and improving water quality. In
conclusion, the high level of compensatory growth capacity, together with the values of PNR50 and PRS50
obtained in this study, could serve as useful indices for assessing brood quality and food quality in
aquaculture.
© 2010 Elsevier B.V. All rights reserved.
1. Introduction
An adequate feeding management in aquaculture is essential to
reduce costs, of which about 50% are devoted to feeding (CortésJacinto et al., 2003a). An important approach to reduce feed costs is to
analyze resistance to temporary food deprivation. The capacity of
crustacean larvae to tolerate periods of starvation is believed to be
essential for their survival in the wild, particularly in unstable habitats
with variable food availability (Anger, 2001).
Anger and Dawirs (1981) demonstrated the existence of the
“point-of-reserve-saturation” (PRS) and the “point-of-no-return”
(PNR) in the larvae of the crab Hyas araneus. The PRS is defined as
the minimum time at which enough reserves are accumulated for
successfully completing a larval stage without food, while PNR is the
threshold at which larvae have lost the capability to recover from the
nutritional stress caused by early starvation, even when subsequently
fed. The PNR was found to occur during the transition between
intermolt and early premolt (Anger, 1987). Knowledge of the critical
points during initial feeding stages may improve larval survival of
⁎ Corresponding author. Fax: +54 11 4576 3384.
E-mail address: [email protected] (L.S. López Greco).
0044-8486/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.aquaculture.2010.03.011
commercially cultured species (Abrunhosa and Kittaka, 1997; Liddy
et al., 2003). In crustaceans, the PNR and PRS can be quantified
experimentally to characterize the “nutritional vulnerability” (Sulkin,
1978) or “nutritional flexibility” (Sulkin and van Heukelem, 1980).
Another potential way to reduce feeding costs is to take advantage
of the process known as compensatory growth, which represents an
increase in weight gain as a result of growth restriction during an
early period of food deprivation (Kim and Lovell, 1995; Eroldogan
et al., 2006). This management tool is extensively applied in fish
farming (Miglavs and Jobling, 1989; Jobling and Kostela, 1996; Gibson
Gaylord and Gatlin, 2001; Ali et al., 2003; Eroldogan et al., 2006;
among others) and could also be used for crustaceans.
The degree of compensatory growth following food deprivation is
highly dependent not only on the species and feeding protocols but
also on the length and intensity of food deprivation (Oh et al., 2008).
Possible responses to food deprivation include no compensation,
partial compensation, full compensation and overcompensation. In
full compensation, the deprived animals eventually achieve the same
size at the same age as continuously fed animals, and in partial
compensation the deprived animals fail to achieve the same size as
nonrestricted animals but show relatively rapid growth rate during
the refeeding period (Ali et al., 2003). These phenomena may have
practical applications in aquaculture, maximizing food efficiency,
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
minimizing food waste and enhancing growth rate (Heide et al., 2006;
Oh et al., 2008).
In crustaceans, most studies of compensatory growth have been
conducted in the marine shrimp Fenneropenaeus chinensis. These
focused mainly on the effects of repetitive starvation–refeeding cycles
(Wu and Dong, 2001), different temperatures (Wu and Dong, 2002a),
protein restriction in terms of body weight percentage and crude
protein percentage (Wu and Dong, 2002b; Wu et al., 2001) and
hypoxic exposure (Wei et al., 2008). The effect of intermittent feeding
was evaluated in the marine shrimp Litopenaeus vannamei (Zheng
et al., 2008) and the freshwater shrimp Macrobrachium nipponense (Li
et al., 2009), which are capable of undergoing compensatory growth.
The freshwater crayfish Cherax quadricarinatus is an omnivorous
species native to the North of Queensland (Australia), and the
Southeast of Papua New Guinea (Jones, 1997), with a fast growth
rate, easy management and high productivity (Cortés-Jacinto et al.,
2003a). It is used worldwide for human consumption and ornamental
purposes (Luchini and Panné Huidobro, 2008).
C. quadricarinatus has been the subject of many studies dealing
with the effect of artificial diets on growth and nutrition (Meade and
Watts, 1995), nutritional requirements (Gu et al., 1996; VillarrealColmenares, 2002; Hernández-Vergara et al., 2003; Cortés-Jacinto et
al., 2003b, 2004a, 2004b, 2005; Thompson et al., 2006; CampañaTorres et al., 2008), spatial distribution of food (Barki et al., 1997) and
feeding frequency (Cortés-Jacinto et al., 2003a). However, all these
studies were limited to pre-adults and adults and no information
is available for early development stages. In species with direct
development like C. quadricarinatus, the first two juvenile stages
are lecithotrophic and exogenous feeding begins at the free-living
stage III (Levi et al., 1999).
The objective of this study was to determine the nutritional
vulnerability of stage III juveniles of C. quadricarinatus based on the
estimation of PSR and PNR, and to evaluate their compensatory
growth capacity when fed intermittently.
2. Materials and methods
2.1. Test animals and experimental conditions
Stage III juveniles were obtained under laboratory conditions from
reproductive stocks supplied by Farm Las Golondrinas, Entre Ríos,
Argentina. Ten ovigerous females (mean wet body weight± SD 62.62±
7.31 g) were placed individually into 30-l glass aquaria (60 × 40× 30 cm).
These contained dechlorinated water (pH 7.5, hardness 80 mg/l as CaCO3
equivalents) under continuous aeration to maintain a dissolved oxygen
concentration of 5 mg/l, and the photoperiod was 14L:10D (Jones, 1997).
Temperature was held constant at 26–27 °C by ALTMAN water heaters
(100 W, accuracy of 1 °C). The females were fed daily ad libitum with
Elodea sp. and commercial balanced food for tropical fish (Tetracolor,
TETRA®), containing 475 g/kg crude protein; 65 g/kg crude fat; 20 g/kg
crude fiber; 60 g/kg moisture; 15 g/kg phosphorus and 100 µg/kg
ascorbic acid. This diet was previously found to be adequate for the
studied species (Vazquez et al., 2008; Sánchez de Bock and López Greco,
2010).
After reaching the free-living stage III (Levi et al., 1999), juveniles
were separated from their mothers and maintained under the
laboratory conditions described above. Juveniles were dried with
paper towel and carefully weighed using an analytical balance
(accuracy of 0.001 g), and then placed in individual plastic containers
(250 cm3) with 200 ml of dechlorinated water and a piece of synthetic
net as shelter (3 × 3 cm) (Sánchez de Bock and López Greco, 2010).
Juveniles were randomly assigned to different feeding protocols of
two experiments: the PRS–PNR assay and the intermittent feeding
assay. In each experiment, 20 replicates were assigned per treatment.
Each replicate consisted of one juvenile placed in a single plastic
container with water and shelter as described above.
35
During these experiments, the plastic containers were cleaned
and water was renewed daily. On feeding days, animals were
offered a nutritionally balanced food (TETRA®) ad libitum once
daily, and checked twice daily (morning and afternoon) for molts
and deaths.
2.2. PRS and PNR experiment
The indices PRS50 and PNR50 were calculated. PRS50 is the time
when 50% of initially fed juveniles are capable of molting to the
following stage and PNR50 is the time when 50% of initially starved
juveniles have lost the capability to molt (Anger and Dawirs, 1981;
Paschke et al., 2004). This experiment consisted of different feeding
and starvation treatments until molting to stage IV; the experimental
period is herein referred to as the restriction period (Fig. 1a).
Five hundred and twenty stage III juveniles (mean initial weight ±
SD 17.23±1.18 mg) were randomly assigned to 26 protocols: 12
feeding treatments identified from F1 to F12, with an increasing
number of feeding days followed by continuous starvation (Fig. 1b),
12 starvation treatments identified from S1 to S12, with an increasing
number of starvation days followed by continuous feeding (Fig. 1b)
and two controls consisting of continuously fed (CF) and continuously
starved (CS) animals. After molting to stage IV, final weight, survival
and duration of stage III were recorded. The experiment ended when
all juveniles molted or died.
2.3. Intermittent feeding experiment
The possible effect of intermittent feeding on compensatory
growth was evaluated in 400 randomly selected stage III juveniles
(mean initial weight ± SD 15.79 ± 1.71 mg). This experiment, which
lasted for 30 days, consisted of alternating periods of feeding and
starvation until molting to stage IV (restriction period), followed by a
refeeding period (Fig. 1a). The juveniles were randomly assigned to
two groups, each of which comprised eight treatments and two
controls, CF and CS. The individuals in one group were initially fed and
those in the other group were initially starved (Fig. 1c).
Each of the following treatments was applied to 20 individuals
of the first group: FS1 (1 day fed/1 day starved), FS2 (2 days fed/
2 days starved), FS3 (3 days fed/3 days starved), FS4 (4 days fed/
4 days starved), FS5 (5 days fed/5 days starved), FS6 (6 days fed/6 days
starved), FS7 (7 days fed/7 days starved), FS8 (8 days fed/8 days
starved) and the controls, CF and CS.
Each of the following treatments was applied to 20 individuals of the
second group: SF1 (1 day starved/1 day fed), SF2 (2 days starved/2 days
fed), SF3 (3 days starved/3 days fed), SF4 (4 days starved/4 days fed),
SF5 (5 days starved/5 days fed), SF6 (6 days starved/6 days fed), SF7
(7 days starved/7 days fed), SF8 (8 days starved/8 days fed) and the
controls CF and CS. In all cases, the restriction period lasted until the
juveniles molted to stage IV. Thereafter, they were continuously fed
until the end of the experiment.
Juveniles were weighed immediately after molting to stage IV, and
again at 15 and 30 days of the experiment. For each treatment, the
survival and number of molts were recorded on days 15 and 30, and
the duration of stage III was determined.
2.4. Calculations and statistical analysis
Survival, expressed as percentage, was calculated at the end of the
restriction period for the PSR–PNR experiment and at days 15 and 30
during the refeeding period of the intermittent feeding experiment
(Fig. 1a).
Molting was calculated as the percentage of individuals in a given
treatment that molted to stage IV. The mean number of molts (NM)
was calculated as the number of molts in a given treatment divided by
the number of individuals that molted. Molting was estimated for
36
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
Fig. 1. Experimental design used in stage III juveniles of Cherax quadricarinatus. (a) General schedule for treatments; (b) protocol applied during the restriction period: feeding
treatments performed to determine the point-of-reserve-saturation (PRS) and starvation treatments performed to determine the point-of-no-return (PNR); (c) protocol applied
during the restriction period: intermittent feeding protocol in the initially fed group and in the initially starved group. For abbreviations see the Materials and methods section.
Fed . Unfed □.
both experiments while NM was only estimated for the intermittent
feeding experiment, on days 15 and 30.
To estimate PRS50 and PNR50, data of molting percentage for the
initial feeding periods or initial starvation periods (feeding or
starvation treatments, respectively) were fit to a sigmoid curve
using the equation f = a / {1 + exp. − [(x − x0) / b]}.
Juvenile's growth was evaluated in terms of growth increment
(GI) expressed as percentage and mean specific growth rate (SGR)
expressed as percentage per day. These were calculated as follows:
GI = 100 × ((W t − W 0) / W0 ) and SGR = 100 × (ln W t − ln W0 ) / t,
where Wt and W0 are final and initial wet weights and t is the
time, estimated as the number of days from the beginning of the
experiment to the first molt and from the first molt to 15 and to
30 days of the experiment. In the PSR and PNR experiment, the GI
was calculated at the end of the restriction period. In the
intermittent feeding experiment, GI and SGR were calculated at
the end of the restriction period and at 15 and 30 days of the
experiment (Fig. 1a).
The independence between survival or molting and treatments
was tested using the Chi-square test of independence followed by
Fisher's test for multiple comparisons between treatments and CF
(Zar, 1999).
The parametric tests were applied when data met the appropriate
assumptions; otherwise, equivalent non-parametric tests were used.
One-way ANOVA or Kruskal–Wallis test (non-parametric) were used
to test for differences in the number of molts, duration of stage III, GI
and SGR among treatments, followed by Dunnett's or Mann–Whitney
(non-parametric) tests for multiple comparisons between treatments
and CF (Zar, 1999).
3. Results
3.1. PRS and PNR experiment
3.1.1. PRS: feeding treatments
Survival depended on the initial feeding period (p b 0.001).
Survival in CS, F1 and F7 differed significantly from that in CF. No
molting was observed in CS and F1, while a large amount of juveniles
achieved molting with two or more days of initial feeding. The
relationship between percentage of molting and initial feeding period
is shown in Fig. 2a (p b 0.001). The estimated value of PRS50 was
3.53 days. In CF, the duration of stage III of juveniles that molted to
stage IV was 9.80 ± 4.06 days and no significant differences (p N 0.05)
were found between treatments and CF (Fig. 2c). The GI obtained for
CF was only significantly higher than that of F2 (Table 1).
3.1.2. PNR: starvation treatments
Survival was dependent on the initial starvation period (p b 0.001),
with lower values in CS, S12, S11, S9 and S7 than in CF. The molting
percentage was lower for all treatments when compared with CF. The
relationship between percentage of molting and initial starvation
period is shown in Fig. 2b (p b 0.001). The estimated value of PNR50
was 4.28 days. The juveniles showed a tendency to delay molting with
increasing number of starvation days. There were significant
differences in the duration of stage III among treatments (p b 0.001),
with S4, S5, S6, S7 and S8 being significantly longer than CF (Fig. 2d).
The treatments S9, S10, S11 and S12 were not included in the analysis
because of their high mortality. No significant differences in GI were
observed among treatments where juveniles molted and CF (p N 0.05).
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
37
Fig. 2. Molting percentage and duration of stage III (mean ± SD) of Cherax quadricarinatus obtained under different protocols. (a) and (c): PRS treatments; (b) and (d): PNR
treatments. For abbreviations see the Materials and methods section. Asterisk (*) indicates significant differences (p b 0.05) between each treatment and the continuously fed control
(CF).
3.2. Intermittent feeding experiment
3.2.1. Initially fed group
Survival at days 15 and 30 of the experiment depended on
the treatment (p b 0.001). On day 15, survival was significantly lower
in FS1 and CS than in CF. On day 30, CS showed no survivors, while
survival in FS1 and FS5 was significantly lower than in CF (Fig. 3a).
Table 1
Mean growth increment (GI, %) and standard deviation (SD) of juveniles of Cherax
quadricarinatus during the PRS and PNR experiment.
CF
GI (%)
SD
37.5
17.1
Feeding treatments
F1
F2
F3
F4
F5
F6
F7
F8
F9
F10
F11
F12
–
14.1a
35.9
30.8
34.0
40.4
47.5
37.9
42.8
34.9
43.2
38.7
CS
GI (%)
SD
–
–
Starvation treatments
–
3.5
12.5
14.6
15.1
17.3
17.2
11.5
21.2
12.0
7.9
14.3
S1
S2
S3
S4
S5
S6
S7
S8
S9
S10
S11
S12
27.9
24.8
37.5
30.0
26.3
27.4
20.5
22.0
15.6
31.5
12.6
30.9
14.1
13.9
17.2
16.0
7.5
13.6
8.3
18.5
16.5
15.0
9.5
6.5
a
Indicates significant differences (p b 0.05) between each treatment and the
continuously fed control (CF).
Molting to stage IV depended on the treatment (p b 0.001). Only
CS, where no molts occurred, was significantly different from CF
(Fig. 4a). The duration of stage III differed significantly among
treatments (p = 0.004), with FS3 being longer than CF (Fig. 5a). No
significant differences in the mean number of molts (NM) were found
among treatments at 15 and 30 days of the experiment (Table 2).
At the end of the restriction period, GI was significantly different
among treatments (p = 0.001), with a significantly lower value for FS1
compared with CF (Fig. 6a). Likewise, SGR also varied significantly
among treatments (p = 0.002), with a significantly lower value for FS1
compared with CF (Table 2). No significant differences in GI and SGR
were obtained among treatments at days 15 and 30 of the experiment
(p N 0.05) (Fig. 6a, Table 2).
3.2.2. Initially starved group
In this group, survival at days 15 and 30 of the experiment also
depended on the treatment (p b 0.001). On day 15, survival in CS, SF5,
SF6 and SF8 was significantly lower than in CF. At the end of the
experiment, CS and the treatments with 6 or more days of initial
starvation showed no survivors, and survival in SF5 was significantly
lower than in CF. In contrast, survival was significantly higher in SF2
than in CF (Fig. 3b).
No molts were observed in CS, SF6 and SF8 and only two
individuals molted in SF7. Molting to stage IV depended on the
treatment (p b 0.001). Molting percentages in SF1, SF4, SF5 and SF7
were significantly different from CF (Fig. 4b). The duration of the stage
III was significantly different among treatments (p = 0.003), with SF5
and SF7 being significantly longer than CF (Fig. 5b). On day 15, the NM
38
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
Fig. 3. Survival percentage of juveniles of Cherax quadricarinatus during the
intermittent experiment: (a) initially fed group; (b) initially starved group. For
abbreviations see the Materials and methods section. Asterisk (*) indicates significant
differences (p b 0.05) between each treatment and the continuously fed control (CF).
Fig. 5. Duration of stage III (mean ± SD) of Cherax quadricarinatus in the intermittent
experiment (a) initially fed group; (b) initially starved group. For abbreviations see the
Materials and methods section. Asterisk (*) indicates significant differences (p b 0.05)
between each treatment and the continuously fed control (CF).
Table 2
Specific growth rate and number of molts of juveniles of Cherax quadricarinatus during
the intermittent experiment (mean ± SD). Only individuals that molted at least once
were considered.
Restriction period
Refeeding period
SGRa
SGR
NMb
SGR
NM
Initially fed group
CF
6.0 ± 2.7
FS1
2.6 ± 1.5c
FS2
4.9 ± 2.7
FS3
4.9 ± 1.9
FS4
5.5 ± 2.8
FS5
5.7 ± 2.0
FS6
6.3 ± 2.0
FS7
5.7 ± 2.1
FS8
6.3 ± 2.2
5.8 ± 0.9
5.8 ± 1.0
5.5 ± 1.3
6.0 ± 0.3
5.2 ± 1.4
5.8 ± 0.6
5.2 ± 1.3
5.4 ± 1.1
5.3 ± 1.0
1.4 ± 0.5
1.6 ± 0.5
1.5 ± 0.5
1.5 ± 0.4
1.2 ± 0.4
1.6 ± 0.4
1.6 ± 0.5
1.3 ± 0.5
1.4 ± 0.5
4.1 ± 0.9
3.6 ± 0.3
3.8 ± 1.0
3.6 ± 1.4
3.8 ± 1.5
4.2 ± 0.6
3.8 ± 0.8
4.1 ± 1.6
4.2 ± 0.8
2.4 ± 0.9
2.8 ± 0.5
2.1 ± 0.6
2.2 ± 0.8
2.2 ± 0.6
2.8 ± 0.4
2.7 ± 0.5
2.3 ± 0.9
2.8 ± 0.6
Initially starved group
CF
6.3 ± 2.2
SF1
4.1 ± 1.6c
SF2
2.9 ± 0.9c
SF3
2.5 ± 0.9c
SF4
2.4 ± 1.2c
SF5
2.7 ± 0.7c
SF6
–
SF7
1.9 ± 1.3c
SF8
–
3.0 ± 2.2
3.6 ± 2.7
2.7 ± 2.3
2.5 ± 1.8
3.6 ± 2.9
3.9 ± 1.2
–
ns
–
1.4 ± 0.5
1.4 ± 0.5
1.1 ± 0.2c
1.0 ± 0.0c
1.5 ± 0.5
1.0 ± 0.0c
–
ns
–
3.4 ± 1.0
3.6 ± 0.9
4.1 ± 0.9
4.7 ± 0.9
3.2 ± 0.8
ns
–
ns
–
2.3 ± 0.9
2.8 ± 0.4
2.7 ± 0.6
2.1 ± 0.9
2.3 ± 1.2
ns
–
ns
–
Treatments
Fig. 4. Molting and mortality percentages of stage III of Cherax quadricarinatus recorded
during the restriction period of the intermittent experiment: (a) initially fed group; (b)
initially starved group. For abbreviations see the Materials and methods section.
Asterisk (*) indicates significant differences (p b 0.05) between each treatment and the
continuously fed control (CF).
ns indicates no survival.
a
SGR (specific growth rate: % day− 1) = 100 × (ln Wt − ln W0) / t, where Wt and W0
are final and initial wet weights and t is the time in days.
b
NM (mean number of molts) = number of molts in a given treatment divided by
the number of individuals that molted.
c
Indicates significant differences (p b 0.05) between each treatment and the
continuously fed control (CF).
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
Fig. 6. Percentage of growth increment (mean ± SD) of the juveniles of Cherax
quadricarinatus during the restriction and refeeding periods of the intermittent
experiment: (a) initially fed group; (b) initially starved group. For abbreviations see
the Materials and methods section. Asterisk (*) indicates significant differences
(p b 0.05) between each treatment and the continuously fed control (CF).
differed significantly among treatments (p = 0.001), with lower
values for SF2, SF3 and SF6 than for CF. In contrast, no significant
differences in NM were found among treatments (p N 0.05) on day 30
(Table 2).
At the end of the restriction period, GI and SGR differed
significantly among treatments (p = 0.001). The GI in all treatments
was significantly lower than in CF, except for SF5 (Fig. 6b) and the SGR
in all treatments was significantly lower than in CF (Table 2). No
significant differences in GI and SGR were found among treatments at
days 15 and 30 of the experiment (p N 0.05).
4. Discussion
The results of the PRS-feeding treatments indicated that stage III
juveniles of C. quadricarinatus required 2 days of initial feeding to molt
to the next stage, but that 3 days of initial feeding were necessary to
reach a weight comparable to that of the continuous feeding control.
The PRS50 estimated for these juveniles was 3.53 days representing
36% of the stage duration, which is consistent with the information
provided by Anger and Dawirs (1981) for larvae of H. araneus. The
authors pointed out that about 50% of the larvae could reach the next
stage when energy reserves were stored for at least one third of the
stage duration. In comparison, the phyllosoma larvae of the lobster
Panulirus cygnus required a higher percentage of the stage duration
(42%) (Liddy et al., 2003) and the zoea I of the shrimp C. crangon a
lower percentage, for both winter (23%) and summer (32%) larvae
(Paschke et al., 2004).
The PNR50 obtained for stage III juveniles of C. quadricarinatus was
4.28 days, which represents 44% of the stage duration. This percentage
is lower than those reported previously, e.g. 54% for P. cygnus (Liddy et
al., 2003), more than 50% for H. araneus (Anger and Dawirs, 1981) and
39
more than 70% for C. crangon (Paschke et al., 2004). In all starvation
treatments, the molting percentage was lower than and the GI similar
to those of the continuously fed control. In addition, development of
the juveniles was prolonged from 2 days of initial starvation onward.
This tendency toward a delayed molting with increasing time of initial
starvation has also been observed in other crustaceans (Anger et al.,
1981; Anger, 1995a; Liddy et al., 2003; Paschke et al., 2004; Calado et
al., 2008; Figueiredo et al., 2008).
To our knowledge, the results of the PRS–PNR experiment
presented here are the first reported for a subtropical freshwater
crayfish. C. quadricarinatus would be expected to exhibit high
starvation resistance due to the features of its habitat. Jalihal et al.
(1993) found a tendency toward a reduction in the number of larval
stages and an increase in larval starvation resistance when comparing
prawns of the genus Macrobrachium, and assumed that it resulted
from a gradual evolution during the “freshwaterization process”. A
similar pattern was observed in crabs of Grapsoidea as part of the
conquest of freshwater habitats (Anger, 1995b).
The present study demonstrates that stage III juveniles of C.
quadricarinatus are unable to molt to the next stage without feeding.
García-Guerrero et al. (2003) reported that the first two stages after
egg hatching (juveniles I and II in the present paper) showed an
increasing rate of protein intake, probably related to higher energy
demand for differentiation and growth. This may explain the
importance of feeding at stage III, after yolk has been exhausted. In
the crab Sesarma curacaoense, the zoeae I and II (first stages after
hatching) show facultative lecithotrophy; they initially reveal large
quantities of yolk, but when food is available, the larvae will
accumulate additional energy reserves enhancing the starvation
resistance of later stages (Anger, 1995a, 1995b). In C. quadricarinatus,
the fact that I and II stage juveniles are lecithotrophic, may drive freeliving juveniles III to need more feed for reaching the next stage
earlier, when compared to the other studied crustaceans.
In the intermittent feeding experiment, stage III juveniles of C.
quadricarinatus showed full growth compensation depending on the
number of days of food deprivation, the sequence of feeding days and
whether the juveniles were initially fed or starved. Juveniles initially
fed for 2 or more days before starvation showed similar growth, in
terms of both GI and SGR, to that of continuously fed juveniles. On the
other hand, juveniles fed and starved for alternating 1-day periods
(FS1) had 3 feeding days during the restriction period and were
significantly lighter than the control, suggesting that the amount of
energy stored was less than that required for normal growth. In
contrast, the weight of those that were continuously fed for 3 days
during the restriction period of the PRS–PNR experiment was similar
to that of the control.
During the refeeding period of the intermittent experiment, all
juveniles in the initially fed group showed a growth similar to that of
the control, but those in FS1 had lower survival. Different results were
obtained for the initially starved group, where full compensation was
achieved only during the refeeding period, as indicated by values of GI
and SGR similar to those of the control. At molting to stage IV, all
treatments showed lower GI and SGR than the control, except for
those starved and fed for alternating periods of 5 days (SF5).
The treatments SF1, SF2, SF3 and SF4, which exceeded the PRS50
value of 3.53 days, had 4 feeding days before molting to stage IV. This
suggests the importance of not only the number of feeding days but
also the moment at which feeding begins because stage III juveniles
would accumulate energy reserves at the first days of the stage. On the
other hand, the treatments SF5 to SF8 showed no molting or small
percentages of molting and survival, probably explained by the PNR50
value of 4.28 days. In addition, juveniles in these treatments showed a
tendency toward a delayed molting, as previously mentioned.
Summarizing, both the FS and SF groups showed full growth
compensation, with the former compensating in the restriction period
and the latter in the refeeding period. In addition, the number of molts
40
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
in both FS and SF groups was similar to that in the control at day 30 of
the experiment. These responses may reflect an adaptation to
changing environmental conditions at early stages of development
as part of the “freshwaterization” process (Jalihal et al., 1993).
The adequate use of compensatory growth at early developmental
stages of C. quadricarinatus can improve feed efficiency (Wang et al.,
2000), since comparatively high yields can be obtained with a lower
number of feeding days. Gu et al. (1996) reported an important decrease
in the mean body weight of C. quadricarinatus juveniles (130.6 mg)
following deprivation of food for 12 days; they gained weight as soon as
they began feeding, but with 6 days of refeeding, which was a short
period, they could not reach the same control weight.
F. chinensis juveniles (mean body weight 2.2 g) showed partial
compensatory growth when exposed to different durations of food
deprivation and refeeding periods (Wu and Dong, 2001) and juveniles
of that species (mean body weight 486 mg) also demonstrated full
compensatory growth when exposed to different temperatures (18 to
26 °C) (Wu and Dong, 2002a). By contrast, the postlarvae of
L. vannamei of about 150 mg fed and fasted for alternating 12-day
periods exhibited little or no growth in comparison with those fed for
24 days (Stuck et al., 1996). The juveniles of M. nipponense (mean
weight 580 mg) failed to compensate when starved for 8 days
followed by refeeding during 10 days (Li et al., 2009).
In conclusion, despite the relatively short period of evaluation
(30 days), stage III juveniles of C. quadricarinatus kept under
intermittent feeding had growth and survival values similar to those
obtained with the protocol of continuous feeding currently used in
culture farms. The immediate advantage of this is a reduction in the
quantity of food, leading to lower costs of production and improved
water quality conditions. Further studies are needed to determine
whether the benefits of the intermittent feeding regime extend to
development stages beyond stage III. On the other hand, the level of
compensatory growth capacity, together with the values of PNR50 and
PRS50 obtained in this study, could serve as useful indices for assessing
brood quality and food quality in aquaculture.
Acknowledgements
This work is part of a scholarship awarded to NSC by CONICET,
Argentina, and the postgraduate theses of LS and NSC (University of
Buenos Aires, Argentina). This research was funded by Agencia
Nacional de Promoción Científica y Tecnológica (PICT 2007, project
01187), UBACYT (X 458) and CONICET (PIP 00129). We are grateful to
Carlos Anselmi (Las Golondrinas Farm) for providing the reproductive
stock and to the anonymous reviewers whose constructive comments
improved this manuscript.
References
Abrunhosa, F.A., Kittaka, J., 1997. Effect of starvation on the first larvae of Homarus
americanus (Decapoda, Nephropidae) and phyllosomas of Jasus verreauxi and J.
edwardsii (Decapoda, Palinuridae). Bull. Mar. Sci. 61, 73–80.
Ali, M., Nicieza, A., Wootton, R.J., 2003. Compensatory growth in fishes: a response to
growth depression. Fish Fish. 4, 147–190.
Anger, K., 1987. The D0 threshold: a critical point in the larval development of decapod
crustaceans. J. Exp. Mar. Biol. Ecol. 108, 15–30.
Anger, K., 1995a. Starvation resistance in larvae of semisterrestrial crab, Sesarma
curacaoense (Decapoda: Grapsidae). J. Exp. Mar. Biol. Ecol. 187, 161–174.
Anger, K., 1995b. The conquest of freshwater and land by marine crabs: adaptations in
life-history patterns and larval bioenergetics. J. Exp. Mar. Biol. Ecol. 193, 119–145.
Anger, K., 2001. The biology of decapod crustacean larvae. Crustacean Issues, 14. A.A.
Balkema, Lisse, The Netherlands. 420 pp.
Anger, K., Dawirs, R.R., 1981. Influence of starvation on the larval development of Hyas
araneus (Decapoda: Majidae). Helgol. Meersunters 34, 287–311.
Anger, K., Dawirs, R.R., Anger, V., Costlow, J.D., 1981. Effects of early starvation periods
on zoeal development of brachyuran crabs. Biol. Bull. 161, 199–212.
Barki, A., Levi, T., Shrem, A., Karplus, I., 1997. Ration and spatial distribution of feed
affect survival, growth, and competition in juvenile red-claw crayfish, Cherax
quadricarinatus, reared in the laboratory. Aquaculture 148, 169–177.
Calado, R., Dionisio, G., Bartilotti, C., Nunes, C., Santos, A., Dinis, M.T., 2008. Importance
of light and larval morphology in starvation resistance and feeding ability of newly
hatched marine ornamental shrimps Lysmata spp. (Decapoda: Hippolytidae).
Aquaculture 283, 56–63.
Campaña-Torres, A., Martínez-Córdova, L.R., Villarreal-Colmenares, H., Civera-Cerecedo, R., 2008. Carbohydrate and lipid digestibility of animal and vegetal
ingredients and diets for the pre-adult redclaw crayfish, Cherax quadricarinatus
(von Martens). Aquac. Res. 39, 1115–1121.
Cortés-Jacinto, E., Villarreal-Colmenares, H., Rendón-Rumualdo, M., 2003a. Efecto de la
frecuencia alimenticia en el crecimiento y sobrevivencia de juveniles de langosta de
agua dulce Cherax quadricarinatus (von Martens, 1868) (Decapoda: Parastacidae).
Hidrobiológica 13, 151–158.
Cortés-Jacinto, E., Villarreal-Colmenares, H., Civera-Cerecedo, R., Martínez-Córdova,
L., 2003b. Effect of dietary protein level on growth and survival of juvenile
freshwater crayfish Cherax quadricarinatus (Decapoda: Parastacidae). Aquac.
Nutr. 9, 207–213.
Cortés-Jacinto, E., Villarreal-Colmenares, H., Civera-Cerecedo, R., Naranjo-Páramo, J.,
2004a. Effect of dietary protein level on the growth and survival of pre-adult
freshwater crayfish Cherax quadricarinatus (von Martens) in monosex culture.
Aquac. Int. 35, 71–79.
Cortés-Jacinto, E., Villarreal-Colmenares, H., Civera-Cerecedo, R., Cruz-Suárez, L.E.,
2004b. Studies on the nutrition of the freshwater crayfish Cherax quadricarinatus
(von Martens): effect of the dietary protein level on growth of juveniles and preadults. Freshwater Crayfish 14, 70–80.
Cortés-Jacinto, E., Villarreal-Colmenares, H., Cruz-Suárez, L.E., Civera-Cerecedo, R.,
Nolasco-Soria, H., Hernández-Llamas, A., 2005. Effect of different dietary protein
and lipid levels on growth and survival of juvenile Australian redclaw crayfish,
Cherax quadricarinatus (von Martens). Aquac. Nutr. 11, 283–291.
Sánchez de Bock, M., López Greco, L.S., 2010. Sex reversal and growth performance in
juvenile females of the freshwater crayfish Cherax quadricarinatus (Parastacidae):
effect of increasing temperature and androgenic gland extract in the diet. Aquac.
Int. 18, 231–243.
Eroldogan, O.T., Kumlu, M., Kiris, G.A., Sezer, B., 2006. Compensatory growth response
of Sparus aurata following different starvation and refeeding protocols. Aquac.
Nutr. 12, 203–210.
Figueiredo, J., Penha-Lopes, G., Narciso, L., Lin, J., 2008. Effect of starvation during late
megalopa stage of Mithraculus forceps (Brachyura: Majidae) on larval duration,
synchronism of metamorphosis, survival to juvenile, and newly metamorphosed
juvenile size. Aquaculture 274, 175–180.
García-Guerrero, M., Racotta, I.S., Villarreal-Colmenares, H., 2003. Variation in lipid,
protein, and carbohydrate content during the embryonic development of the
crayfish Cherax quadricarinatus (Decapoda: Parastacidae). J. Crust. Biol. 23, 1–6.
Gibson Gaylord, T., Gatlin, D.M., 2001. Dietary protein and energy modifications to
maximize compensatory growth of channel catfish (Ictalurus punctatus). Aquaculture 194, 337–348.
Gu, H., Anderson, A.J., Mather, P.B., Capra, M.F., 1996. Effects of feeding level and
starvation on growth and water and protein content in juvenile redclaw crayfish,
Cherax quadricarinatus (von Martens) Mar. Fresh. Res. 47, 745–748.
Heide, A., Foss, A., Stefansson, S.O., Mayer, I., Norberg, B., Roth, B., Jenssen, M.D.,
Nortvedt, R., Imsland, A.K., 2006. Compensatory growth and fillet crude
composition in juvenile Atlantic halibut: effects of short term starvation periods
and subsequent feeding. Aquaculture 261, 109–117.
Hernández-Vergara, M.P., Rouse, D.B., Olvera-Novoa, M.A., Davis, D.A., 2003. Effects of
dietary lipid level and source on growth and proximate composition of juvenile
redclaw (Cherax quadricarinatus) reared under semi-intensive culture conditions.
Aquaculture 223, 107–115.
Jalihal, D.R., Sankolli, K.N., Shenoy, S., 1993. Evolution of larval developmental patterns
and the process of freshwaterization in the prawn genus Macrobrachium Bate, 1868
(Decapoda, Palaemonidae). Crustaceana 65, 365–376.
Jobling, M., Kostela, J., 1996. Interindividual variations in feeding and growth in
rainbow trout during restricted feeding and in a subsequent period of compensatory growth. J. Fish Biol. 49, 658–667.
Jones, C.M., 1997. The biology and aquaculture potential of the tropical freshwater
crayfish Cherax quadricarinatus. Department of Primary Industries, Queensland.
Information Series, Q190028. Queensland Department of Primary Industries,
Brisbane. 109 pp.
Kim, M.K., Lovell, R.T., 1995. Effect of restricted feeding regimens on compensatory
weight gain and body tissue changes in channel catfish Ictalurus punctatus in ponds.
Aquaculture 135, 285–293.
Levi, T., Barki, A., Hulata, G., Karplus, I., 1999. Mother-offspring relationships in the redclaw crayfish Cherax quadricarinatus. J. Crust. Biol. 19, 477–484.
Li, Z.H., Xie, S., Wang, J.X., Sales, J., Li, P., Chen, D.Q., 2009. Effect of intermittent
starvation on growth and some antioxidant indexes of Macrobrachium nipponense
(De Haan). Aquac. Res. 40, 526–532.
Liddy, G.C., Phillips, B.F., Maguire, G.B., 2003. Survival and growth of instar 1 phyllosoma
of the western rock lobster, Panulirus cygnus, starved before or after periods of
feeding. Aquac. Int. 11, 53–67.
Luchini, L., Panné-Huidobro, S., 2008. Perspectivas en Acuicultura: nivel mundial, regional
y local. Dirección de Acuicultura; Secretaría de Agricultura, Ganadería, Pesca y
Alimentos (SAGPyA), Subsecretaría de Pesca y Acuicultura, Argentina. 98 pp.
Meade, M.E., Watts, S.A., 1995. Weight gain and survival of juvenile crayfish Cherax
quadricarinatus fed formulated feeds. J. World Aquac. Soc. 26, 469–474.
Miglavs, I., Jobling, M., 1989. Effects of feeding regime on food consumption, growth
rates and tissue nucleic acids in juvenile Artic charr, Salvelinus alpinus, with
particular reference to compensatory growth. J. Fish Biol. 34, 937–957.
Oh, S.-Y., Noh, C.H., Kang, R.-S., Kim, C.-K., Cho, S.H., Jo, J.-Y., 2008. Compensatory
growth and body composition of juvenile black rockfish Sebastes schlegeli following
feed deprivation. Fish. Sci. 74, 846–852.
L. Stumpf et al. / Aquaculture 304 (2010) 34–41
Paschke, K.A., Gebauer, P., Buchholz, F., Anger, K., 2004. Seasonal variation in starvation
resistance of early larval North Sea shrimp Crangon crangon (Decapoda:
Crangonidae). Mar. Ecol. Prog. Ser. 279, 183–191.
Stuck, K.C., Watts, S.A., Wang, S.Y., 1996. Biochemical responses during starvation and
subsequent recovery in postlarval Pacific white shrimp, Penaeus vannamei. Mar.
Biol. 125, 33–45.
Sulkin, S.D., 1978. Nutritional requirements during larval development of the portunid
crab, Callinectes sapidus Rathbun. J. Exp. Mar. Biol. Ecol. 34, 29–41.
Sulkin, S.D., van Heukelem, W.F., 1980. Ecological and evolutionary significance of
nutritional flexibility in planktotrophic larvae of the deep sea red crab Geryon
quinquedens and the stone crab Menippe mercenaria. Mar. Ecol. Prog. Ser. 2, 91–95.
Thompson, K.R., Metts, L.S., Muzinic, L.A., Dasgupta, S., Webster, C.D., 2006. Effects of
feeding practical diets containing different protein levels, with or without fish
meal, on growth, survival, body composition, and processing traits of male and
female Australian red claw crayfish (Cherax quadricarinatus) grown in ponds.
Aquac. Nutr. 12, 227–238.
Vazquez, F.J., Tropea, C., López Greco, L.S., 2008. Development of the female
reproductive system in the freshwater crayfish Cherax quadricarinatus (Decapoda,
Parastacidae). Inv. Biol. 127, 433–443.
Villarreal-Colmenares, H., 2002. Avances en Nutrición Acuícola. In: Cruz-Suarez, LE.,
Ricque-Marie, D., Tapia-Salazar, M., Gaxiola-Cortés, M.G., Simoes, N. (Eds.),
Memorias del VI Simposium Internacional de Nutrición Acuícola. Cancún, Quintana
Roo, México, pp. 114–142.
41
Wang, Y., Cui, Y., Yang, Y., Cai, F., 2000. Compensatory growth in hybrid tilapia
Oreochromis mossambicus × O. niloticus, reared in seawater. Aquaculture 189,
101–108.
Wei, L.-Z., Zhang, X.-M., Li, J., Huang, G.-Q., 2008. Compensatory growth of Chinese
shrimp, Fenneropenaeus chinensis following hypoxic exposure. Aquac. Int. 16,
455–470.
Wu, L., Dong, S., 2001. The effects of repetitive “starvation-and-refeeding” cycles on the
compensatory growth response in Chinese shrimp, Fenneropenaeus chinensis
(Osbeck, 1765) (Decapoda: Penaidae). Crustaceana 74, 1225–1239.
Wu, L., Dong, S., 2002a. Compensatory growth responses in juvenile chinese shrimp,
Fenneropenaeus chinensis, at different temperatures. J. Crustac. Biol. 22, 511–520.
Wu, L., Dong, S., 2002b. Effects of protein restriction with subsequent realimentation on
growth performance of juvenile Chinese shrimp (Fenneropenaeus chinensis).
Aquaculture 210, 343–358.
Wu, L., Dong, S., Wang, F., Tian, X., Ma, S., 2001. The effect of previous feeding regimes
on the compensatory growth response in Chinese shrimp, Fenneropenaeus
chinensis. J. Crust. Biol. 21, 559–565.
Zar, J.H., 1999. Biostatistical Analysis, 4th ed. Prentice-Hall Inc., New Jersey, USA.
663 pp.
Zheng, Z., Dong, S., Tian, X., 2008. Effects of intermittent feeding of different diets on
growth of Litopenaeus vannamei. J. Crust. Biol. 28, 21–26.