FISHERIES SCIENCE 2002; 68: 465–477

Effect of different photoperiod cycles on metabolic rate

and energy loss of both fed and unfed young tilapia
Oreochromis niloticus: Part I
Amal Kumar BISWAS, Masato ENDO AND Toshio TAKEUCHI*

Department of Aquatic Biosciences, Tokyo University of Fisheries, Minato,

Tokyo 108-8477, Japan

ABSTRACT: The influence of different photoperiod cycles (3L : 3D, 6L : 6D, 12L : 12D, and
24L : 24D) on the metabolic rate and energy loss of either fed or unfed young tilapia Oreochromis
niloticus (bodyweight 8.6–9.5 g) was investigated at 28∞C. A computer-operated respirometer with a
closed tank was used to prevent water from evaporating into the air or condensing from the air. The
photoperiods acted as strong Zeitgeber (i.e., cue or synchronizer) during the experiments with either
fed or unfed fish. A photoperiod-mediated metabolic cycle was demonstrated in fed and unfed fish
in which oxygen consumption was higher during the light phase compared with during the dark phase
for all photoperiods. Mean oxygen consumption during the 3L : 3D, 6L : 6D, 12L : 12D, and 24L : 24D
periods for fed and unfed fish were 685.06 mg/kg per h and 299.33 mg/kg per h; 658.52 mg/kg per h
and 284.80 mg/kg per h; 591.09 mg/kg per h and 249.62 mg/kg per h; and 500.64 mg/kg per h and
239.14 mg/kg per h, respectively. The highest post-prandial increase in energy loss was recorded
during the 3L : 3D period (145.88 kJ/kg per day), followed by 141.19 kJ/kg per day during the 6L : 6D,
128.70 kJ/kg per day during the 12L : 12D, and 99.92 kJ/kg per day during the 24L : 24D periods. The
results suggest that higher energy conservation would be achieved if fish are exposed to longer rather
than shorter photoperiod cycles.

KEY WORDS: energy loss, Oreochromis niloticus, oxygen consumption, photoperiod,

Zeitgeber

INTRODUCTION been adopted by a number of investigators.10,11

One of the main reasons for the wide distribu-
tion of Oreochromis niloticus around the world,
mostly in tropical and subtropical regions, is its
ability to tolerate large variations in environmen-
tal factors.1,2 A number of investigators have mea-
sured the metabolic rates of different species of
tilapia. Routine metabolic rates have been mea-
sured by Ross and McKinney,3 Caulton,4,5 Mishrigi
and Kubo,6 and Ross and Ross;7 and the metabolic
requirements of feeding fish have been docu-
mented by a number of investigators also.8,9 To
study the post-prandial increase in oxygen con-
sumption by teleosts, a quantitative approach has
*Corresponding author: Tel/Fax: 81-3-5463-0545. Email:
take@tokyo-u-fish.ac.jp
Received 11 May 2001. Accepted 31 October 2001.
They have investigated the effects of different
feeding levels on the magnitude of the post-
prandial increase in oxygen consumption during a
natural photoperiod.
The metabolic rate in fishes, as measured by
oxygen consumption, can be influenced by a large
number of internal and external factors.12 Temper-
ature and size,5,13–15 various types of pollution,16
season,17 and activity18 have been examined in
some detail. Some investigators have reported a
varying metabolic rate that is linked to daily pho-
toperiod,14,19 and it may be influenced by longer-
term changes in photoperiod.20 Among the various
environmental factors, photoperiod is one of the
main artificial Zeitgebers (i.e. cue or synchronizer)
that may influence the daily rhythm of fish. In
fishes, light has various effects on circadian varia-
tion in some physiological activities.21–23
466 FISHERIES SCIENCE
Little research has been conducted on the influ-
ence of different photoperiod regimens on the
metabolic rate and energy loss of either fed or
unfed O. niloticus. To date, all of the studies that
have been undertaken have used either a natural
photoperiod (i.e. 12L : 12D), or a longer light phase
and shorter dark phase or vice versa. Any variation
from the 24-h light/dark cycle may influence the
adaptation of the endogenous rhythm to the exter-
nal cycle.24 Therefore, it would seem quite likely
that the adaptation of the endogenous rhythm
to an external cycle might influence the physiolo-
gical processes, including those related to animal
growth rate. Thus, if one animal lives through
many more cycles than another, but in the same
timespan, it may use energy differentially and
grow at a different speed. However, the higher
post-prandial increase in metabolic rate or energy
loss doesn’t always act to reduce the ‘scope for
activity’. It may also resemble an inescapable ‘cost
of growth’. The present paper is one of a series on
the influence of different photoperiod regimens on
the growth of O. niloticus, as determined using an
energetic model, and describes the effect of differ-
ent photoperiod regimens on the metabolic rate
and energy loss of fed and unfed young tilapia.
MATERIALS AND METHODS
It is common practice in a flow-through respirom-
eter to measure dissolved oxygen (DO) levels
before and after the respirometer chamber,3,5,25,26
which requires the use of at least two polaro-
graphic electrodes and much servicing and cali-
bration by attendants. The closed recirculatory
system that was used in the present experiment
provided controlled environmental conditions
that varied little during the entire experiment.
The study was conducted using a closed-fish rear-
ing tank (SAS-2; Toyo Engineering Corp., Chiba,
Japan), which is constructed to establish a Closed
Ecological Recirculating Aquaculture System
(CERAS).27 This closed system was used to prevent
water from evaporating into the air or condensing
from the air. The system consisted of a 55-L closed
tank with sensors (two sensors for DO and one
for both pH and temperature), a settling tank,
two filters (a biofilter and a coral filter), a circula-
tory pump, a CO2–O2 exchange unit, and a heater.
Various components (e.g. settling tank, filters,
CO2–O2 exchange unit, etc.) provide controlled
environmental conditions that maintain some
chemical components within ranges that are
tolerable to fish.27 A schematic diagram of the
closed-fish rearing tank and water flow is shown in
Fig. 1.
AK Biswas et al.
Physical parameters of the system
Culture tank
A 55-L, airtight, cylindrical culture tank (Toyo Engi-
neering Corp.) was used. It was made of acrylic
resin and measured 400 mm in diameter and was
500 mm deep. A feeding port and temperature
sensor was located on the upper side of the tank.
Circulated water entered the tank from the upper
side and moved out by a pipe located at the base of
the tank.
Settling tank
The cylindrical settling tank (Toyo Engineering
Corp.) was also made of acrylic resin but measured
130 mm in diameter and was 350 mm depth,
having a water volume of 3.5 L. The circular move-
ment of water permitted the heavy solid particles
to settle directly onto the bottom of the tank. A
wool filter was placed on the upper side of the tank
to reduce particulate materials in the system.
Biofilter and coral sand tank
The biofilter and coral sand tank were also both
cylindrical, acrylic resin tubes, holding water
volumes of 5.5 L and 1.5 L, respectively. The biofil-
ter, containing porous sintered glass beads/siporax
(Schott, Tokyo, Japan), was used for nitrification of
ammonia produced by the fish. The coral sand
tank, containing coral sand and filter wool, stabi-
lized the system’s pH. The biofilter and coral sand
tank both reduced particulate materials in the
system and helped to prevent levels of metabolites
from becoming too high.
Water supply system (pump)
A 100 V–38 W water supply pump (model MD
15FX-N; Iwaki Co. Ltd, Tokyo, Japan) was used to
circulate the water. The magnetic impeller of the
pump was able to maintain the flow rate at
1–5 L/min through its OD 16 ¥ ID 12 mm silicon
rubber tubing supply line.
O2–CO2 Exchange unit (oxygenator)
The oxygenator is a cylindrical, hollow fiber mem-
brane module (model KMF 011 HZS; Mitsubishi
Rayon Co. Ltd, Tokyo, Japan) measuring 70 mm in
diameter, 220 mm in length and 2.4 m2 in mem-
brane area. Gas exchange was achieved by the
continuous introduction of air through the O2–CO2
Oxygen consumption and energy loss in young tilapia: I FISHERIES SCIENCE 467

Personal
Feeding computer
hole

Buffer
tank Rearing tank
55 L Data sending
Temperature
28°C Water flow

Sensor
[DO( PiO2) & pH]

Settling
Sensor tank
DO(P0O2) 3.5 L
Heater
(Temperature control) Coral sand tank
(pH control)

CO2/O2 exchange unit 1.5 L

Biofilter
5.5 L
Air flow
Pump

Fig. 1 Diagram of the fish-rearing closed tank. Arrow indicates the direction of water flow.

exchange unit. This unit helped to keep dissolved to control the pressure and to compensate the
oxygen near the saturation level. water volume.

Heating system
A 100 V–300 W inline heater (Aqua Co. Ltd, Tokyo,
Japan) was installed in the water flow line to main-
tain the system’s water temperature within ± 0.2∞C.
Buffer tank
A 1.2-L buffer tank (Nalgene Corp., Tokyo, Japan)
was connected to the base pipe of the culture tank
Illumination
Different periods of light and dark were controlled
by an automatic timer (F5S Time Switch; Omron
Corp., Kyoto, Japan) switch connected to a
100 V–20 W fluorescent tube (inverter). It changed
gradually at 30 min intervals to achieve the light
and dark phases.
Sensors (dissolved oxygen, pH, and temperature)
The DO of water was automatically measured by a
468 FISHERIES SCIENCE
DO sensor (GV-BRM5; Iijima Electronics Corp.,
Tokyo, Japan) and was converted to a proportional
voltage using a digital DO meter (model MC-7W-S;
Iijima Electronics Corp.) with appropriate circuitry.
The pH was automatically measured by a pH meter
suspended in the water flow path and was con-
verted into a proportional voltage using a digital
pH meter (model MC-7P; Iijima Electronics Corp.).
In the same way, temperature was automatically
read by a resistance thermometer (class B, 2 mA)
and was converted into a proportional voltage
using a digital temperature indicator (Fenwal Con-
trols of Japan Ltd, Tokyo, Japan) with appropriate
circuitry.
Similarly, flow rate and time were converted
to a proportional voltage. The voltage outputs
from these sensors were stored automatically in a
computer.
During operation, the water flowed from the
O2–CO2 exchange unit to the fish-rearing closed
tank, through the first DO sensor and pH sensor to
the settling tank, through the second sensor for DO
to the biofilter, to the coral sand tank, to the pump,
and back around the loop.
Measurement of oxygen consumption
Genetically pure O. niloticus of Ibaraki strain were
used for various photoperiod combinations. To
empty their alimentary canals, all tested individu-
als were starved for 48 h before the experiment was
commenced. The fish were then anesthetized
with 100 p.p.m. 4-ethylaminobenzoate (Wako Pure
Chemical Industries Ltd, Osaka, Japan) and
weighed before placing them in the closed tank.
The fish were allowed to acclimate to the system’s
environment for approximately 24 h before the
experiment was commenced. The system’s opera-
tion was checked as described earlier, and the fish
were then monitored over a 5-day feeding period.
Fish were fed a commercial diet (crude protein
50.7%, crude lipid 9.3%; Nippon Formula MFG Co.
Ltd, Yokohama, Japan) at a level of 3% bodyweight,
four times a day during the light phase of all exper-
imental photoperiods, with the exception of the
24L : 24D experiment, during which the fish were
fed on alternate days. Two replicates of each
photoperiod experiment was carried out. Approxi-
mately 20 fish (average bodyweight 8.6 g) were
stocked in each tank to measure the metabolic rate
in fed small fish. For the unfed state, 30 fish
(average bodyweight 9.5 g) were starved for 6 days,
except for the 24L : 24D experiment, during
which the starvation duration was only 5 days.
Four types of photoperiods (i.e. 3L : 3D, 6L : 6D,
12L : 12D, and 24L : 24D) were studied and the
AK Biswas et al.
oxygen consumption between the various fish
groups were compared. The feeding schedule and
various parameter types used during the different
photoperiod cycles are given in Fig. 2 and Table 1,
respectively.
At the end of each experiment, data were
collected from the computer and reprocessed.
Energy expenditure was calculated using an indi-
rect calorimetry method; that is, it was calculated
by converting the daily oxygen consumed to
energy, using the oxycalorific coefficient of
13.68 J/mg O24 for unfed tilapia and 14.85 J/mg O2
for fed tilapia,28 because the respiratory quotient
for unfed and fed fish is different.28
Statistical analysis
anova was performed to identify any differences in
oxygen consumption between the duplicate exper-
iments and among the various treatments. Data
were then subjected to Tukey’s test with a 95%
significance level to compare the means when
differences occurred.
RESULTS
Oxygen consumption in fed fish
High oxygen consumption during both the light
and dark phases was observed in the shorter pho-
toperiod cycles (e.g. 3L : 3D and 6L : 6D) compared
with the longer photoperiod cycles (12L : 12D and
24L : 24D) (Fig. 3). Furthermore, differences in
oxygen consumption were observed between the
light and dark phases and vice versa (Fig. 4). In the
fed fish experiments, there was no significant dif-
ference among the 3L : 3D, 6L : 6D and 12L : 12D
photoperiods (P > 0.05); however, the 3L : 3D and
6L : 6D photoperiods showed a significant differ-
ence in oxygen consumption compared with the
24L : 24D photoperiod (P < 0.05) during the light
phase. During the dark phase, the 3L : 3D photope-
riod showed a significant difference with the
12L : 12D and 24L : 24D photoperiods. The dark
phases of the 6L : 6D and 12L : 12D photoperiods
also showed a significant difference compared with
the 24L : 24D photoperiod (Fig. 3). Figure 5 shows
the cyclic metabolic rhythm in which the peaks of
metabolic activity coincide with daylight and the
troughs coincide with darkness for all photoperi-
ods. The amplitude of the cycles was higher for
long photoperiod cycles than for short photope-
riod cycles (Fig. 5).
Oxygen consumption and energy loss in young tilapia: I FISHERIES SCIENCE 469

3L:3D

0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48

6L:6D

6 12 18 24 30 36 42 48
0

12L:12D

Fig. 2 Feeding schedule 0 12 24 36 48

during different photoperiod
cycles. Arrow indicates the time 24L:24D
of feeding and the black bar
shows the dark phase of the
photoperiod. 0 24 48

Table 1 Rearing conditions of fed (n = 40) and unfed (n = 60) Oreochromis niloticus during the experiment
Feeding experiment Unfed experiment
Parameters 3L : 3D 6L : 6D 12L : 12D 24L : 24D 3L : 3D 6L : 6D 12L : 12D 24L : 24D
No. fish 40 40 40 40 60 60 60 60
Duration 5 5 5 5 6 6 6 5
(days)
Average 8.56 ± 0.96 8.56 ± 0.91 8.54 ± 0.82 8.56 ± 0.86 9.50 ± 1.07 9.52 ± 1.12 9.50 ± 0.98 9.51 ± 1.02
bodyweight
(g)*
Feeding 3 3 3 3 – – – –
(% bodyweight)
Water flow 3.84 ± 0.07 3.79 ± 0.09 3.80 ± 0.08 3.77 ± 0.07 3.90 ± 0.11 3.78 ± 0.10 3.79 ± 0.10 3.82 ± 0.11
(L/min)†
Temperature 27.8 ± 0.09 27.9 ± 0.09 27.9 ± 0.08 27.9 ± 0.07 27.9 ± 0.07 27.9 ± 0.08 27.9 ± 0.09 27.9 ± 0.08
(∞C)†
pH† 7.39 ± 0.08 7.40 ± 0.09 7.42 ± 0.12 7.37 ± 0.06 7.41 ± 0.10 7.39 ± 0.06 7.40 ± 0.10 7.42 ± 0.08

*Values are mean ± SD.

†
Mean ± SD (n = 20).

Oxygen consumption in unfed fish
Data for fishes subjected to various photoperiod
cycles were pooled and averaged into 1-h blocks.
The reprocessed data is shown in Fig. 6. Although
the fish were starved throughout the experiment in
order to evaluate oxygen consumption of tilapia in
an unfed state, their metabolic cycles were main-
tained until the end of each photoperiod cycle
without any apparent decrement (Fig. 6). The less
apparent rhythmicity of the 3L : 3D and 6L : 6D
photoperiods compared with the 12L : 12D and
24L : 24D photoperiods might be due to the shorter
light/dark phase cycle of the former photoperiods.
The peaks of metabolic activity coincide with day-
light and the troughs coincide with darkness for
all photoperiods. The shorter photoperiod cycles
showed higher routine oxygen consumption
(referred to as oxygen consumption at resting or
unfed state) than the longer photoperiod cycles
(Fig. 3). In contrast, a higher difference in oxygen
consumption was observed between the light and
dark phases of the longer photoperiod cycles than
of the shorter photoperiod cycles (Fig. 4). In this
experiment the 3L : 3D photoperiod was signifi-
cantly different to the 12L : 12D and 24L : 24D
periods during both the light and dark phases
(P < 0.05).
When fish were exposed to a continuous light
phase, the metabolic rhythm remained cyclic over
the entire period (Fig. 6). The peaks and troughs of
the plotted data continued to correspond closely
470 FISHERIES SCIENCE AK Biswas et al.

900
(a) routine metabolic rate for all photoperiod cycles,
800 can be made. An example of the procedure used to
700 estimate biomass change is given:
600
500
Number of fish = 30
Period of starvation = 6 days
Oxygen consumption (mg/kg per h)

400 AB
A A B
300
X XY Y Z
Photoperiod cycle = 3L : 3D
200 Mean actual loss in biomass of group
100
0
(n = 30) = 732 mg
3L : 3D 6L : 6D 12L : 12D 24L : 24D Average weight of group during starvation = 9.50 g
Average condition factor (CF) of group during
400
(b) starvation = 2.96
350
Total metabolic energy requirement for this group
300
of fish in the 3L : 3D photoperiod over a period
250 of 6 days = Average weight of group ¥ oxygen con-
200 sumed in 3L : 3D photoperiod ¥ oxy-energy coeffi-
A AB B B
150 cient ¥ 6 days
100
X XY YZ Z = 9.50 g ¥ 7.18 (mg/g per day) ¥ 13.68 (J/mg O2) ¥ 6
50 = 5602 J.
0 The ratio of energy utilized by a fish with a CF of
3L : 3D 6L : 6D 12L : 12D 24L : 24D 2.96 has been estimated as 3.2 fat units to 1 protein
Photoperiods unit;4 or 76.19% of the energy would be derived
from fat, whereas 23.81% would be derived from
Fig. 3 Oxygen consumption in: (a) fed; and (b) unfed protein. Thus, 4268 J would be derived from fat
Oreochromis niloticus during the () light and () dark metabolism and 1334 J from protein. Because the
phases during applied photoperiods. Values given are energy content of fat is 39.54 kJ/g and protein is
the mean ± SD (fed n = 40; unfed n = 60). Bars show
23.64 kJ/g,29 then the mass of fat needed to give
oxygen consumption during the light and dark phases,
whereby those sharing the same letters are not signifi- 4268 J would be 108 mg and the mass of protein
cantly different (P > 0.05). needed would be 56 mg. Thus, the total mass
requirement of 5602 J would be 164 mg. This esti-
mated organic biomass loss must be added to the
corresponding water loss component, which for a
fish with a CF of 2.96 can be estimated as 3.5 units
Oxygen consumption (mg/kg per h)

120 of water for every unit of tissue.4 Hence, for a loss

100 of 164 mg of fat and protein, an estimated equiva-
80 lent loss of water would be 574 mg, giving a total
60 biomass loss of 738 mg. This theoretical value is in
agreement with the mean actual loss of 732 mg.
40
Following the same procedure, comparisons were
20
made for the 6L : 6D, 12L : 12D, and 24L : 24D pho-
0 toperiod experiments, and these results are tabu-
3L : 3D 6L : 6D 12L : 12D 24L : 24D
lated in Table 2.
Photoperiods

Fig. 4 Differences in oxygen consumption between the

light and dark phases for fed ( , oxygen consumed Post-prandial metabolic rate and energy loss
during dark phase is deducted from that consumed
during the light phase) and unfed ( , same as for fed The increased rate of oxygen consumption
fish) fish. induced by feeding appears to be approximately
double that of the ‘routine/standard metabolic
rate’ for all photoperiod cycles tested. The
highest increase was observed for the 12L : 12D
with the photoperiod cycle to which the fish had photoperiod (2.37 times), followed by the 6L : 6D
been acclimated. The rhythmicity and amplitude photoperiod (2.31 times), the 3L : 3D photoperiod
of variation was suppressed when fish were (2.29 times), and the 24L : 24D photoperiod
exposed to continuous darkness (Fig. 6). (2.09 times). However, the mean increase (amount
Comparisons between changes in biomass, as of oxygen consumed by unfed fish deducted from
measured using fish starved for 6 days, and a the- amount of oxygen consumed by fed fish) of oxygen
oretical biomass change, as calculated using the consumption caused by feeding showed higher
Oxygen consumption and energy loss in young tilapia: I FISHERIES SCIENCE 471

(a)
1200
3L : 3D

1000

800
Oxygen consumption (mg/kg per h)

600

400

200

0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57 60 63 66 69 72 75 78 81 84 87 90 93 96 99 102 105 108 111 114 117 120

1200
6L : 6D
1000

800

600

400

200
0 6 12 18 24 30 36 42 48 54 60 66 72 78 84 90 96 102 108 114 120

Time (h)

(b)
1000
12L : 12D

800

600
Oxygen consumption (mg/kg per h)

400

200
0 12 24 36 48 60 72 84 96 108 120

1000
24L : 24D

800

600

400

200
0 24 48 72 96 120

Time (h)

Fig. 5 (a) Variation in oxygen consumption of fed Oreochromis niloticus during photoperiods 3L : 3D and 6L : 6D. (b)
Variation in oxygen consumption of fed O. niloticus during photoperiods 12L : 12D and 24L : 24D. Black bar at the
bottom of the figure indicates the dark phase of the photoperiod cycles. () Respiration during the light phase of the
photoperiod; () respiration during the dark phase of the photoperiod. Circles and triangles indicate duplicate data.
472 FISHERIES SCIENCE AK Biswas et al.

(a)
550
3L : 3D

450

350
Oxygen consumption (mg/kg per h)

250

150

50
0 3 6 9 12 15 18 21 24 27 30 33 36 39 42 45 48 51 54 57 60 63 66 69 72 75 78 81 84 87 90 93 96 99 102 105 108 111 114 117 120 123 126 129 132 135 138 141 144

550
6L : 6D

450

350

250

150

50
0 6 12 18 24 30 36 42 48 54 60 66 72 78 84 90 96 102 108 114 120 126 132 138 144
Time (h)

(b)
450
12L : 12D

350

250
Oxygen consumption (mg/kg per h)

150

50
0 12 24 36 48 60 72 84 96 108 120 132 144

450
24L : 24D

350

250

150

50
0 24 48 72 96 120
Time (h)

Fig. 6 (a) Variation in oxygen consumption of unfed Oreochromis niloticus during photoperiods 3L : 3D and 6L : 6D.
(b) Variation in oxygen consumption of unfed O. niloticus during photoperiods 12L : 12D and 24L : 24D. Black bar at the
bottom of the figure indicates the dark phase of the photoperiod cycles. () Respiration during the light phase of the
photoperiod; () respiration during the dark phase of the photoperiod. Circles and triangles indicate duplicate data.
Oxygen consumption and energy loss in young tilapia: I FISHERIES SCIENCE 473

Table 2 Comparison of measured and estimated change in mass during starvation at different photoperiod cycles*
Photoperiod 3L : 3D 6L : 6D 12L : 12D 24L : 24D
No. fish 30 30 30 30
Time of starvation (day) 6 6 6 5
Condition factor (CF)† 2.96 2.96 2.97 2.98
Total measured mass lost (mg/30 fish) 21970 21020 15040 14440
Estimated mass of tissue used (mg/30 fish) 4920 4710 4110 3930
Estimated mass of water lost (mg/30 fish) 17220 16500 14400 13770
Total estimated mass lost (mg/30 fish) 22140 21210 18510 17700

*Average value (n = 2)
†
Condition factor (CF) = 100 ¥ W/L3; where W is weight (g) and L is length (cm).

600 350
Oxygen consumption (mg/kg per h)

500 300

Energy loss (kJ/kg per day)

400
300
200
100
0
3L : 3D
Fig. 7 Post-prandial increase in oxygen consumption
rate by Oreochromis niloticus during the light (,
amount of oxygen consumed by unfed fish is subtracted
from the amount of oxygen consumed by fed fish during
the light phase) and dark (, amount of oxygen con-
sumed by unfed fish is subtracted from the amount of
oxygen consumed by fed fish during the dark phase)
phases during different photoperiods. Values given are
mean ± S.D (fed n = 40; unfed n = 60).
values (385.73 mg/kg per h) in the 3L : 3D photope-
riod, followed by 373.72 mg/kg per h in the 6L : 6D
photoperiod, 341.47 mg/kg per h in the 12L : 12D
photoperiod, and 261.50 mg/kg per h in the
24L : 24D photoperiod (Fig. 7).
The computed value (by the subsequent con-
version of the daily oxygen consumed to energy
using the oxycalorific coefficient of 13.68 J/mg O2
for unfed tilapia and 14.85 J/mg O2 for fed tilapia28)
of energy loss for fed and unfed fish, and the
increase in post-prandial energy loss (energy loss
by unfed fish subtracted from that lost by fed fish)
are given in Fig. 8. The 3L : 3D photoperiod cycle
showed the highest energy expenditure in the fed
and unfed fish and also demonstrated the highest
post-prandial increases, followed by the 6L : 6D,
12L : 12D, and 24L : 24D photoperiods. The energy
loss due to metabolism in fed and unfed fish
exposed to the 3L : 3D, 6L : 6D, 12L : 12D, and
250
200
150
100
50
6L : 6D 12L : 12D 24L : 24D 0
3L : 3D 6L : 6D 12L : 12D 24L : 24D
Photoperiods
Photoperiods
Fig. 8 Energy loss by ( ) fed and ( ) unfed Ore-
ochromis niloticus; and ( ) post-prandial increase in
energy loss during the relevant photoperiod. Values
given are mean ± S.D (fed n = 40; unfed n = 60).
24L : 24D photoperiod cycles were 244.16 kJ/kg per
day and 98.28 kJ/kg per day; 234.70 kJ/kg per day
and 93.51 kJ/kg per day; 210.66 kJ/kg per day and
81.96 kJ/kg per day; and 178.43 kJ/kg per day and
78.51 kJ/kg per day, respectively. The post-prandial
increase in the 3L : 3D, 6L : 6D, 12L : 12D, and
24L : 24D photoperiods were 145.88 kJ/kg per day,
141.19 kJ/kg per day, 128.70 kJ/kg per day, and
99.92 kJ/kg per day, respectively.
DISCUSSION
The present study was conducted using the closed
recirculating aquaculture system. The overall
objective of this is to establish a closed ecological
recirculating system for aquatic organisms
such as phytoplankton, zooplankton and fish,
and to adapt it for fish culture in space. At
present, many experiments are being conducted to
improve our knowledge of rearing conditions,
culture techniques, etc. The present study was con-
474 FISHERIES SCIENCE
Table 3 Comparison of routine metabolic rate for a range of tilapia species at 28∞C
Routine metabolic rate (mg/kg per h)
Species Photoperiod
Oreochromis mossambicus 12L : 12D
12L : 12D
12L : 12D
12L : 12D
Tilapia rendalli 12L : 12D
Oreochromis niloticus 3L : 3D
6L : 6D
12L : 12D
24L : 24D
ducted with the aforementioned goal in mind. The
study’s results, coupled with the results from an
elaborate growth study that applies these pho-
toperiod regimens, will help in determining the
optimum photoperiod for rearing fish in this
system.
It has been noted by many investigators that the
metabolic rate of fish varies with time,3,30 and this
was clearly evident in the present work, which
studied various photoperiod cycles (Figs 5,6). The
daily trends in the data may be resolved into either
a single daily peak3,30 or a crepuscular pattern.31,32
In the present study, the metabolic rate sometimes
showed a single peak for O. niloticus, and it was
mostly obscured due to the influence of feeding
during peak activity.
In the feeding experiment, the fish exposed to
the 12L : 12D cycle showed a robust daily rhythm
throughout the recording period. In the other pho-
toperiod cycles, feeding and different photoperiod
cycles acted as a stimulus for synchronizing an
endogenous rhythm with the external environ-
ment, resulting in the establishment of a robust
rhythm that coincided with the respective pho-
toperiod cycle.
In the experiment of oxygen consumption in
unfed fish, a clear metabolic rhythmicity was
observed for all photoperiod cycles, with the
exception of the 3L : 3D cycle, during which rhyth-
micity was less apparent. This may be due to the
shorter frequency of the light and dark phases,
which might not have been enough to cause a
visible rhythmicity. Regardless, the metabolic rate
of unfed fish during this photoperiod was higher in
the light phase than in the dark phase. The higher
rates of metabolic activity observed during the
light phase was probably due to the fish exhibit-
ing behavioral interactions in the light, which
increased their activity levels and therefore their
metabolic rate. Thus, it appears that the biorhythm
AK Biswas et al.
10 g 50 g 80 g Reference
– – 306 Job33
340 238 – Mironova34
– 182 – Caulton4
300 182 155 Caulton5
– 246 – Caulton4
299 – – Present study
285 – – Present study
250 – – Present study
239 – – Present study
is endogenous in origin and is influenced by the
level of light and by the photoperiod, which is in
agreement with the results of Ross and McKinney.3
The metabolic data for O. niloticus obtained by
various investigators is compared in Table 3; only
values derived for the routine metabolic rate (at
resting or unfed state) at 28∞C are used. Although
reason for the differences is unclear, the differ-
ences are partly attributed to differences in the
developmental stage. Also, the respirometer tech-
nique employed and the different photoperiod
cycle lengths could be very important.
Using a general equation derived from studies of
Oreochromis mossambicus,5 the routine metabolic
expenditure of the present study agreed strongly
with that calculated by Hepher et al.35 The
observed energy loss of fasting red tilapia (94 kJ/kg
per day)35 was more or less similar to the results
observed in the present study. However, Meyer-
Burgdorff et al. have observed an overestimate of
metabolic expenditure in unfed fish and an under-
estimate in fed fish.9 The most important factors
responsible for these differences are water temper-
ature, duration of fasting, amount of feeding, fish
species and size, and fish activity. In the present
study, the 3L : 3D and 6L : 6D cycles showed meta-
bolic rates that were 16% and 11% higher for fed
fish, and 20% and 14% higher for unfed fish,
respectively, compared with the 12L : 12D cycle. A
possible explanation is the frequent rotation of the
light and dark phases that occurs during such a
short photoperiod cycle, which may accelerate the
physiological activities. Behavioral interaction,
which was mentioned earlier, or the influence
of the stimulus activity of Zeitgeber, such as the
photoperiod, for synchronizing an endogenous
rhythm to the external environment may also
require more energy in the shorter photoperiod
cycles. But this conclusion is not definite; to obtain
a better understanding, other variables, such as
Oxygen consumption and energy loss in young tilapia: I FISHERIES SCIENCE 475

Table 4 Comparison between feeding and routine metabolic rate in a series of species
Species Photoperiod Relationship with ‘feeding rate’ Reference
Blennius pholis 12L : 12D 1.75 ¥ ‘low routine’ Vahl & Davenport36
Pleuronectes platessa 12L : 12L 2.00 ¥ ‘low routine’ Jobling & Davies37
Kuhlia sandvicensis 12L : 12D 2.60 ¥ ‘standard’ Muir & Niimi38
Micropterus salmoides 12L : 12D 1.78 ¥ ‘standard’ Beamish39
Gadus morhua 12L : 12D 1.60 ¥ ‘routine’ Saunders8
Brevoortia tyrannus 12L : 12D 2.00 ¥ ‘routine’ Hettler40
Lepomis macrochirus 12L : 12D 2.00 ¥ ‘routine’ Schalles & Wissing41
Oreochromis niloticus 3L : 3D 2.29 ¥ ‘routine’ Present study
6L : 6D 2.31 ¥ ‘routine’ Present study
12L : 12D 2.37 ¥ ‘routine’ Present study
24L : 24D 2.09 ¥ ‘routine’ Present study

hormone rhythm or locomotor activity, and
whether or not they are altered by different pho-
toperiod cycles, need to be studied.
A comparison of the post-prandial increase in
metabolic rate in a series of fish species is summa-
rized in Table 4. Investigators have reported a
number of components that influence the post-
prandial oxygen consumption, including intestinal
activity,42 postabsorptive activity,43 and diet com-
position.37,44 However, Schalles and Wissing41 were
unable to detect any differences in the influence of
dietary composition on post-prandial metabolic
rate. Moreover, all fish subjected to different pho-
toperiod cycles were offered the same food; hence,
the influence of diet composition was not consid-
ered in the present study. A close link between
growth and post-prandial increase in metabolic
rate has been demonstrated by a number of inves-
tigators.45,46 Jobling and Davies have shown that
under conditions that give the most rapid rate of
growth, the rate of oxygen consumption is greatest
when it is above the level required for mainte-
nance.37 The higher post-prandial increase in fish
exposed to shorter photoperiod cycles compared
with fish exposed to longer photoperiod cycles
may be consistent with the aforementioned find-
ings and also with the findings of Jobling,47 who
found that the most rapidly growing fish showed
the greatest post-prandial increase in metabolic
rate.
The results of the present study demonstrated
that fish under natural photoperiod (12L : 12D)
conditions showed higher energy conservation
than those exposed to shorter photoperiod cycles
(3L : 3D and 6L : 6D). As mentioned earlier, find-
ings on the relationship between post-prandial
increase in metabolic rate and growth strongly
reinforces the necessity to carry out an elaborate
growth study of this fish under well-controlled, dif-
ferent photoperiod cycles to compare the growth
and feed efficiency among various photoperiod
cycles. Clearly, this would be of immense benefit to
aquaculturists.
ACKNOWLEDGMENTS
The expenses of the present study were defrayed
in part by a Grant-in-Aid for Scientific Research
(No. 10460143) from the Ministry of Education,
Science, Sport, and Culture and by ‘Ground
Research Announcement for Space Utilization’
promoted by Japan Space Forum.
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