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Aqwacwlture Sci. 62 e) , 3Lg - 322 (2074)
Short Paper
Effect of Water Temperature on
Embryonic Development of Yellowfin
Tuna Thunnus albacares Inhabiting
the Eastern Pacific Ocean
Angel Gurr-r-ÉN1, Tomoki HoNRyo2'*, Juan IneRnel,
Amado CaNol, Daniel MAnGuLros3,
Vernon P Scuolof, Jeanne B. WnxroR3,
Maria S. SrBrN3, Toru KoseyesHra
and Yoshifumi S¿we¡e2
Abstract Effect of water temperature on embryonic
development of yellor,vfin tuna was investigated. Eggs
were incubated at different water temperatures:22, 24,
26C (Experiment-1) arrd 28, 30, 32C (Experiment-2).
Developmental condition was observed and the elapsed
time from the 2 or 4 cell stages until hatching was mea-
sured. Hatching occurred at 18.58 h for eggs incubated
at 32'C. Eggs incubated at 22T stopped development
after the morula (late cleavage) stage. Experiment-3
was designed to examine the relationship between
hatching time and the incubation temperature, and was
expressed by two regression lines which intersected at
269:C.
Key words: Yellowfin tuna; Embryonic development;
Water temperature
Yellorvfin tuna (YFT) Tkunnus albacares inhabits the
mixed layer of all the warm seas of the world except
the Mediterranean Sea, and two stocks are recognized
in the Pacific Ocean: the stock of the Eastern Pacifc
Ocean (EPO) and of the Western-Central Pacific Ocean
(WCPO) (I-ehodey and l.eroy 1999). Although a com-
mercially important species, information is limited to
studies in captivi§z on the reproductive activities and
the effects of physical variables on spawning dynam-
ics or early life stage development in YFT (Margulies
et al. 2007). Water temperature and salini§r are factors
which have a major effect on the development of marine
teleosts (Kawahara et al. 1997). Margulies etal. (2007)
estimated the egg stage duration of YFT at water tem-
peratures between 23.3 and 30.0t, and also reported
the stage-specifc egg and early larval development
of YFT at 27'C, where embryonic-stage duration was
21.65h. Wexler et al. (2011) further described the rela-
tionship of the egg stage duration of YFT and mean
incubation temperatures between 20 and 33C as an
inverse quadratic function; within this range they found
the stage duration to be significantly longer at mean
incubation temperatures lower than 22t. Masuma
(2013) investigated the effect of different incubation
temperatures on YFT in Japan and indicated that an
average evaluation index such as the temperature char-
acteristics of Arrhenius's and Higurashi and Tauchi's
form4lfa ( p, a) and values of Q1s constants are much
highe) in YFT inhabiting the WCPO than in Pacifc
bluefin tuna @BT) T. orient¡ilis. However an extensive
study of the temperature effect on the stage-specific
embryonic development of eggs has not yet been con-
ducted in YFT inhabiting the EPO.
In the present study, the effect of temperature on
embryonic development of YFTwas explored as a basis
for early life history sfudies for resource management
and for aquaculture. In the experiments, we observed
several stages of embryonic development at different
water temperatures and recorded the elapsed time until
hatching. The results were compared with embryonic
development of PBT (Miyashita et al. 2000). In addi-
tion, interspecific comparisons were made between
YFT inhabiting the WCPO (Vlasuma 2013) and the EPO
in order to describe the characteristics ofboth indepen-
dent stocks.
The Inter-American Tropical Tuna Commission
established the Achotines laboratory in the Republic
of Panama. A group of 15 wild-caught YFT broodstock
were collected in coastal waters in the ücini§r of the
Achotines Laboratory during 2008-2010 (1.6-4.6 years
old), and were reared in an indoor concrete circular
tank of 1,300 m3. This broodstock has spawned almost
year-round on a near-daily basis. Fertilized eggs were
collected shortly after spawning in the broodstock
tank during June, 2011. The eggs were carefully rinsed
in a plasüc bucket and transferred to 20 /-volume
Received 24 July 2013; Accepted 20 June 2014.
1 Aquatic Resources Authoriff of Panama, Panama 0819-05850, Republic of Panama.
2 Fisheries I¿boratories of Kinki University, Kushimoto, Wakayama 649-3633, Japan.
3 Inter-American Tropical Tuna Commission, l¿ Jol1a, California 92037 1508, USA.
a Faculty of Agriculture, Kinki University, Nakamachi, Nara 631-8505, Japan.
*Corresponding author: Tel, (+81) 735-65-0501; Fax, (+81) 735-65-0974;Emarl,t.honryo@kindaisuiken.jp (T. Honryo).
320 A. Gui11én, T Honryo, J. Ibarra, A. Cano, D. Margulies, V. P Scholey, J. B. Wexle¡ M. S. Stein, T Kobayashi and Y. Sawada
Table 1. Embryonic developmental stage of yellorvfintrnaThunnus albacares and elapsed time (HH:MM) until hatching under different
incubation temperatures
Developmental stages
Incubation temperature (C )Equation between time and incubation temperature
T = a+b á +c 62+d 0:)
(T = time: hr., á = incubation temperature: C)
3028
ZO2422
2 cell
4 cell
16 cel1
Morula (late Cleavage)
Early gastrula
Appearance of embryonic shield
Appearance of Kupffer's vesicle
Beginning of heart beat
First hatch
Hatching
ono
0: 50
1 :40
0:00 0:00
0:30 0:30
1:20 1:00
6:00 5:15
8:15 7:35
72:00 10:00
20:30 16:00
25:75 19:45
29:15 22:00
0:00 0:00 0:00
0:25 0:25 0:20
0:35 0:35 0:35
1: 10 1: 10 1:05
3:50 3:35 3:20
5:35 5:20 4:35
B:35 7:20 6:45
14:05 72:05 11:05
17:50 16 20 L5:20
20:50 20:05 18:35
T = 51.909-5.5550 +0.7987 02-0.002403 (12 = 0.950)
T = 60.814-6.3140 +0.221802-0.002603 02 = O.gtg¡)
T = -85.02+11.4990-0.464302+0.006á3 ¡r2 = O.Ot8¡
T = -t26.47+16.1640-0.62602+0.0078á3 (12 = 0.960)
T = 65.301-3.07 48 0 +0.0269 0 2 +0.0004 á 3 02 : t.oo)
T = 477 .32-44.729 0 +1.457 02-0.0161 á 3 02 = 0.973)
T = 939.3-91.7430 +3.05202-0.$403 Q2 = O.gg7)
T : 1845.4-187.680 +6.431202-0.0n403 02 = O.gg4)
experimental tanks. These tanks were provided moder-
ate aeration during the experiment from a single dif-
fuser. In Experiment-l (Exp-1), fertilized eggs were
obtained at a water temperature of 28.7T and at 28.5C
in Experiment-2 (Exp-2), respectively. Other eggs were
used to estimate the normal hatching rates measured
in up-flowing 100 /-volume cylindrical tanks and the
hatching rates were 98.9E" (28.5T) in Exp-1 and g2.6%o
(28.8C) in Exp-2, respectively.
A total of 6 different temperatures were set (trxy7:22,
24 ard26T;Exp-2:28, 30 and 32C) in order to describe
the embryonic development and the incubation period
until hatching at each temperature. Approximately
1,500 eggs were used for each experimental tank both
in Exp-1 and Exp-2, and the elapsed time from the flrst
observation until each embryonic stage was recorded.
In Exp-l, the developmental stage at collection was
4-ce1l stage, whereas it was 2-ce1l slage in Exp-2. Ten
to 20 eggs were sampled every 5-15 min, and their
development was examined under a microscope using
egg development descriptions of PBT (Miyashita et al.
2000). Determination of egg developmental stage was
based on the stage observed for the majority of eggs
(over 507o) in a single sample.
In Experiment-3, we further investigated the hatch-
ing time of YFT eggs (0.94 t 0.02 mm in diameter and
0.22 mm of oil goblin) at23-32T (at approximately 1t
intervals) in order to estimate the relationship between
incubation temperafure and the time required foom
spawning until hatching by regression analysis. These
eggs were collected in June, 2012 arrd similar methods
of egg collection and incubation protocols of Exp-1 and
2 were used. In addition, temperature characteristics
(¡t, a) were calculated according to Arrhenius's formula
Qt) and Higurashi and Tauchi's (1925) formula;7n D
: K- a 7, where D denotes the time (h), 7 denotes the
temperature (C) and a, K delote the constant values.
Hu¡
9rs
3
22.0 24.0 28.0 30.0 32.0
Temperature (oC)
Fig. 1. Relationship between incubation temperature and embry-
onic developmental stages of yellowdn t:tna Thunnus albacares.
Ga, early gastrula stage; Em, appearance of embryonic shield; Ku,
appearance of Kupffer's vesicle; Hb, beginning of heart beat; Fh;
first hatch; Ha, hatching.
Q1¡ values of constants {efi= (T2/Tlrlto/(0r02)l} were
also calculated in order to compare these values with
those of YFT inhabiting the WCPO (Masuma 2013) .
Embryonic stages and elapsed time from the 2- and
4-cell stages until hatching (HH:MM) under differ-
ent incubation temperatureq is presented in Table 1.
At24T, it took 0.5 h to proceed until the 1&cell stage,
20:30 h until the beginning of the heart beat stage,
ard 29:15 h until hatching. Elapsed time until hatch-
ing was shorter at higher incubation temperatures.
The shortest elapsed time until hatching occurred at
32T (18:35h). The acceleration of development at
higher temperatures was much clearer after the gas-
trula stages to beginning of heart beat stage (Fig. 1).
Hatching was observed at 24, 26, 28, 30 and 32C,
Embryonic Development of Yellowfin Tuna 321
20
u16
FD
4
0
however, embryonic development stopped after the
morula stage (late cleavage) at22T. The relationship
between elapsed time until the first hatching and incu-
bation temperafure was expressed by the regression
equation: ? = 0.2008 0 2 * tz.gzt 0 + 207.g99 1r2 = 0.94s8,
n = l7), where 7 represents time until first hatching
(h), and 0 represents incubation temperature (t)
(Fig.2). As a result of converting the elapsed time at
the completion of hatching into natural logarithm, two
linear regression lines were obtained (Fig. 3). These
two linear regression lines intersected at 26.9C and the
coefficients l-0.7077 1r2 = 0.9++8, n: 6), - 0.0288 (r2
= 0.667, n = 6)l were found to be significantly different
(ANCOVA, P< 0.01).
J = 0.200812 - 12.321x+ 207,899
X'z= 0.9458
20 22 24 26 28 30 32 34
Temperature ("C)
Fig. 2. Relationship between incubation temperature and elapsed
time until hatching.
22 24 26 28 30 32 34
Temperature ("C)
Fig. 3. Two linear regression lines derived from converting the
elapsed time at the completion of hatching into natural logarithm
and the inflection point of its relationship with incubation tem-
perature.
The temperature characteristics and values of the
constants for the relationship between incubation
temperature and the time required for hatching are
shown in Table 2. A comparison of ¡l a, and Q10 values
showed a tendency for higher values at incubation tem-
peratures of 23.2-26.9T (f¿ : 18,959, a: 0.71, Qn: 2.92)
compared to those for 26.9-32.2T (U:4,986, a: 0.03,
Q1s:1.23).
This study elucidated the stage-specifc develop-
ment of YFT inhabiting the EPO, under wide range of
temperatures. This information is useful in the study of
early life history and aquaculture of funas to estimate
the hatching time andlor calculate spawning time. The
embryonic development of YFT was similar to that of
PBT (Miyashita et al. 2000); however, several differ-
ences were observed. For example, in the final stage of
egg development shortly before hatching, the embryo
covered three quarters of the yolk in YF!, whereas
it coy.ered the yolk completely in PBT Regarding the
relatio\ship belween incubation temperature and
development, Miyashita et al, (2000) reported that the
inflection point in the linear regression of incubation
temperature and hatching time was 25T in PBT The
regression lines intersected at 26.9t in this study for
YFI The existence of an inflection point is related to
enzyme reactions associated with hatching ffasunaga
1988). This inclination of the inflection point at a higher
temperature is possibly explained by the fact that YFT
inhabit warmer water areas than PBT In addition, the
temperature characteristics (p, a) and values of Qro
indicated that these values were higher at tempera-
tures below the inflection point at 26.9"C compared to
those at 26.9-32.2T. which is coincident with YFT
inhabiting the WCPO (Masuma 2013). Therefore, the
developmental speed of YFT inhabiting the EPO sug-
gests that their eggs are also susceptible to lower tem-
peratures. Masuma (2013) further reported that the
average temperature constant values of YFT are much
higher than those of PBT which is also confirmed in
this study. Therefore, these results might confirm some
Table 2. The temperature characteristics (u, a) and values of
constants (Qro) for relationship between incubation temperature
and time required for hatching ofYFT eggs
Incubation^ 2? ,-26.9 26.9-32.2 28.2-32.2
temperature (t)
4,986
0.03
t.23
,¿¿: temperature characteristic of Arrhenius's formula (Higurashi
and Tauchi 1925)
o: temperature characteristic of Higurashi and Tauchi's formula
(Higurashi and Tauchi 1925)
z
j
3
!
a
Qro
18,959
0.11
2.92
9,774
0.05
7.7
y = -0.1077¡+ 5.9113
y=-0.028&x+3.8251
322 A. Guillén, T Honryo, J. Ibarra, A. Cano, D. Margulies, V. P Scholey, J. B. Wexler, M. S. Stein, T Kobayashi and Y. Sawada
differences in distribution areas of PBT and YF f,, and
similarities in distribution of YFT stocks in the EPO
andWCPO.
The embryonic development of YFT from fertil-
izatiot to hatching at 27T was also investigated by
Margulies et al. (2007) and \Mexler et al. (2011) for the
EPO, and these durations were similar to the pres-
ent study (i.e. about 22-23.5h at 26-27'C). On the
other hand, Harada et al. (1980) reported that the
time required for hatching of WCPO YFT was longer
at 26:40-42:00h at an average water temperature of
26.4T. This may be explained by the differences in
the methods for obtaining fertilized eggs Ii. e. artifcial
insemination and lengthy transport of eggs until incu-
bation (Harada et al. 1980) versus natural spawningl.
Therefore, the different methods of fertilization could
affect the developmental speed. The required time
fuom fertilization until hatching of YFT was about 22 h
at 26T in this study. In comparison with other conge-
neric species, egg stage durations of bigeye fiirla T.
obeswswere 30h at 25.5-27.5T ffasutake et al. 1973),
and 24:4h at 26.5T in PBT (Miyashita et al. 2000).
Masuma (2013) reported duration of 20.6 h until the
beginning of hatching at 26.7T for YFT inhabiting the
WCPO, and this duration is similar to that found in
this study (19.8 h). Therefore, YFT may have a slightly
shorter egg-stage duration compared to that of other
Thunnus species.
In conclusion, the rate of embryonic development
of YFT inhabiting the EPO is temperature dependent,
similar to other marine teleosts including other tunas
such as T. obesws, PBT, and YFT inhabiting the WCPO.
However, YFT required a shorter time for hatching
compared to that of other Thwnnws species.
Acknowledgments
The authors would like to thank Dr. Shigeru
Miyashita and Dr. Shukei Masuma for assistance of
this comparative study with Pacific bluefin tuna and
yellor,r.fin tuna and also Mr. Shoji Kibe who coordinated
this study in the Republic of Panama. And the authors
wish to thank the staff of the Achotines Laboratory.
Their kind help was necessary to conduct this study.
This study was supported by the SATREPS program,
assisted bv JSf (Japan Science and Technology) and
JICA (J¿p¿11 International Cooperation Agency).
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