- Email: [email protected]
Determining the appropriate time and hormone administration for induced spawning and egg production of the shoemaker spinefoot rabbitfish, Siganus sutor (Valenciennes, 1835) during the reproductive season
Accepted Manuscript Determining the appropriate time and hormone administration for induced spawning and egg production of the shoemaker spinefoot rabbitfish, Siganus sutor (Valenciennes, 1835) during the reproductive season
Hojjatollah Fourooghifard, Kiuomars Roohani Ghadikolaee, Samuel Allen, Maryam Moezzi, Reza Dehghani, Mohammad Reza Zahedi PII: DOI: Reference:
S0044-8486(19)30164-4 https://doi.org/10.1016/j.aquaculture.2019.05.062 AQUA 634177
To appear in:
aquaculture
Received date: Revised date: Accepted date:
21 January 2019 26 May 2019 26 May 2019
Please cite this article as: H. Fourooghifard, K.R. Ghadikolaee, S. Allen, et al., Determining the appropriate time and hormone administration for induced spawning and egg production of the shoemaker spinefoot rabbitfish, Siganus sutor (Valenciennes, 1835) during the reproductive season, aquaculture, https://doi.org/10.1016/ j.aquaculture.2019.05.062
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT Determining the appropriate time and hormone administration for induced spawning and egg production of the shoemaker spinefoot rabbitfish, Siganus sutor (Valenciennes, 1835) during the reproductive season Hojjatollah Fourooghifarda,* [email protected], Kiuomars Roohani Ghadikolaeea
T
[email protected], Samuel Allenb [email protected], Maryam Moezzia
Persian Gulf and Oman Sea Ecological Research Center, Iranian Fisheries Sciences Research
US
a
CR
Reza Zahedia [email protected]
IP
[email protected], Reza Dehghania [email protected], Mohammad
AN
Institute, Agricultural Research, Education and Extension Organization(AREEO), Bandar Abbas, Iran.
Plant and Environmental Sciences Department, New Mexico State University Agricultural Science
ED
Center, Farmington, New Mexico, USA
M
b
*Corresponding author at: Persian Gulf and Oman Sea Ecological Research Center, Iranian
PT
Fisheries Sciences Research Institute, Agricultural Research, Education and Extension
AC
Abstract
CE
Organization (AREEO), Bandar Abbas, Iran
This study was conducted to determine the appropriate time and hormone administration protocol for induced spawning of the shoemaker spinefoot rabbitfish, Siganus sutor. Four different hormone administration treatments including LHRHa2 hormone at dosages of 10 and 20 μg/kg body weight (BW) (treatment
and
), HCG hormone alone at a dosage of 2000 IU/kg
BW (treatment H) and combined with 5 mg metoclopramide/kg BW (treatment HM) plus the control, 1 ml 0.9% saline water (treatment C) were administered on eight dates in mid-2016
ACCEPTED MANUSCRIPT (April 12, May 1, 8, 15 and 22 and June 1,8 and 15). Females (470.14 ± 8.56 g BW) received the hormone injection twice over a 24-hour interval and all males received HCG hormone at a dosage of 2000 IU/kg BW; hence, each fish was used just once for the injection of hormone. A significantly greater number of spawned fish (70.8 ± 16 %) was observed in treatment L2
T
compared with the other treatments (L1: 16.7 ± 10.8 %; H: 45.8 ± 14 %; HM: 45.8 ± 14 %; and
IP
C: 4.2 ± 0.2 %). Significantly greater numbers of eggs per female (469083 ± 70453 eggs) were
CR
obtained in treatment L2 compared with the other treatments (L1: 103375 ± 48872 eggs; H:
US
294500 ± 74348 eggs; HM: 287458 ± 72856 eggs; and C: 18542 ± 18000 eggs). Cluster analysis showed that the best time for induced spawning of S. sutor was 15 to 22 May and the best
AN
hormone for induced spawning was LHRHa2 at a dosage of 20 µg/kg BW of females. Such information can be helpful in the management of breeding programs for this important fish
M
species.
ED
Keywords:
CE
1. Introduction
PT
Rabbitfish; Siganus sutor; HCG hormone; LHRHa2 hormone; Induced spawning.
AC
Development of commercial aquaculture has led to an increase in the number of aquatic species under domestication efforts (Duarte et al., 2007). One of the important prerequisites for domestication and improvement of a sustainable aquaculture industry is the ability to control the reproductive processes and to obtain high quality of eggs and sperm of fish in captivity (Mylonas et al., 2010). Siganids are generally regarded as good food fishes in spite of their relatively small size, and some species have been cultured because of their herbivorous food habits, rapid growth and economic value (Randall et al.,1997). Artificial reproduction of some species of rabbitfishes
ACCEPTED MANUSCRIPT has been investigated in Siganus gattatus, S. revulatus, S. vemiculatus, S. lineatus and S. canaliculatus from the 1970s (Ayson, 1991; Bryan and Madraisau, 1977; Popper et al., 1976; Lam and Soh, 1975; Popper et al., 1973). There are a few reports about the reproduction biology of this species from various parts of the world (Agembe, 2012; Kamukuru, 2006). Some species
T
of rabbitfish such as S. gattatus can spawn without hormonal treatment, every month throughout
IP
the year (Hara et al., 1986). Induced spawning of rabbitfish has been successfully carried out
CR
using HCG injection (500 IU/ fish or about 2 IU/g body weight) in S. gattatus and S. canaliculatus (Juario et al., 1985; Lam, 1974). Effect of using metoclopramide as a dopamine
US
antagonist has been used to facilitate the induction spawning of some species such as silver carp
AN
(Hypothalamichthys molitrix), bream (Abramis brama L.) and common carp (Cyprnus carpio)(Ashraf and Akar, 2013; Kucharczyk et al., 2005; ). It has been reported that spawning
M
behavior of rabbitfish can be related to lunar periodicity (Rahman et al., 2003; Rahman et al.,
ED
2001; Hara et al., 1986). Some species such as white-spotted spinefoot Siganus canaliculatus, spawn around the new moon from April to June, little spinefoot S. spinus spawn around the new
PT
moon from May to July, the goldlined spinefoot S. guttatus, and the streamlined spinefoot S.
CE
argenteus, spawn around the first quarter moon from June to July (Takemura et al., 2010). A first study about the induced spawning of S. sutor was conducted in Iran in 2005 (Foroughifard et al.,
AC
2007). Although the feasibility of broodstocking and maturation in rabbit fish, was studied by Fourooghifard et al. (2009), there were many unknowns to achieve the acceptable result for induced spawning of this species. Hence, a new study was conducted to determine the appropriate time and hormonal treatment for induced spawning and egg production of this species using LHRHa2 and HCG hormones with or without metoclopramide as a dopamine antagonist.
ACCEPTED MANUSCRIPT 2. Materials and methods 2.1. Design of experiment The experiments were conducted at the Persian Gulf and Oman Sea Ecological Research Institute (PGOSERI), Bandar Abbas, Iran, from March to June 2016. Four different hormonal
T
treatments including LHRHa2 hormone at a dosage of 10 μg/ kg body weight (Treatment L1),
CR
IP
LHRHa2 hormone at a dosage of 20 μg/ kg BW (Treatment L2), HCG hormone at a dosage of 2000 IU/Kg BW alone (treatment H) and combined with 5 mg metoclopramide/kg BW
US
(treatment HM), and a control treatment with 1 ml 0.9% saline solution without any hormone (Treatment C) were performed to survey their effects on induced spawning of Siganus sutor at
AN
the environmental temperature of ~26–30 °C. The LHRHa2 hormone was provided from Ningbo
M
Sansheng Pharmaceutical Co., China and the HCG and metoclopramide were provided from Daru Pakhsh Pharmaceutical Co., Iran. Injections were carried out in mid-2016 on eight different
ED
days ( April 12; May 1, 8, 15 and 22; June 1, 8 and 15). Each treatment had three replications
PT
(one female and one male for each replication), therefore 15 females and 15 males were treated on each day (for a total of 120 females and 120 males during the reproduction season). Each fish
CE
was used just once for the injection of hormone during the experiments. Each pair of injected
AC
fish (1 female and 1 male) was introduced to a 1-m3 separate spawning tank. Since , like other Siganids, S. sutor eggs are adhesive, egg collectors consisting of corrugated fiberglass sheets were placed at the bottom of spawning tanks prior to spawning (Nelson et al., 1992). All the experiments were conducted under natural light period and temperature. 2.2. Broodstocks of Siganus sutor A total of 400 adults of Siganus sutor (387.46 ± 11.73 g body weight) (mean± Se) were collected from the coastal waters of Lavan Island, Iran, and were stocked in 5-m3 fiberglass tanks
ACCEPTED MANUSCRIPT at a density of 8 fish/m3 (3-4 kg/m3) (Duray and Juario, 1988). The fish were reared until their average body weight reached 470.14 ± 8.56 g at the reproduction season. Fishes were fed twice a day with commercial shrimp pellets (40% protein, 5% fat) and crab meat at a rate of 3% body weight (Fourooghifard et al., 2009).
IP
T
2.3. Injection of hormones
CR
The LHRHa2 and HCG powder were diluted in 1 ml 0.9% saline solution just prior to injection. In the treatment HM, the HCG powder was diluted in 1 ml 0.9% saline solution and
US
mixed with 0.5 ml of 10 mg /1ml metoclopramide injection vial. Female fish were distinguished from the males by their swollen and soft abdomens and males were identified by the flowing of
AN
milt through the genital pore after a gentle pressure on their abdomen (Hara et al., 1986).
M
Hormones were injected intramuscularly below the dorsal fin, twice (by 24 hour intervals)
ED
to each female at equal doses, between 8-9 a.m., during April–May, when the environmental temperature was about 26-28 °C (Foroughifard et al., 2007; Marte, 1989). All males were
PT
injected using HCG hormone at a dosage of 2000 IU/kg BW below the dorsal fin at the same
CE
time as the second injections of females (Duray and Juario, 1988). 2.4. Fecundity and hatching rate of eggs
AC
After the spawning of fishes, the egg collectors were removed from the spawning tanks and all eggs were washed and collected onto a sieve and were weighted. Three samples of egg (0.4-0.5 g) were taken from the sieve and the eggs of each sample were counted and the average was obtained. The following formula was used to calculate the Fecundity (Islam et al., 2012). Fecundity (F) = n × G/g, where “n” is the number of eggs in each sample, “G” is the weight of total eggs collected on the sieve, and “g” is the weight of sample
ACCEPTED MANUSCRIPT A batch of 5000 eggs belonging to each female was stocked in a 300-liter round polyethylene tank at a density of 25 eggs per liter to calculate the hatching rate of eggs (Fourooghifard et al., 2017). Hatching rates were obtained by counting the larvae at 1 DAH and using the following formula (Naeem et al., 2011): Hatching rate = 100(Number of larvae/ Total
IP
T
eggs).
CR
2.5. Data analysis
Data were analyzed using SPSS 22.0 (IBM Corp., Armonk, NY, USA) software through
US
the application of parametric tests. The significance of differences was tested by one-way
AN
analysis of variance (ANOVA) and the post-hoc Duncan’s test. Differences were considered significant at p < 0.05. A hierarchical analysis was used for determination of the appropriate time
3. Results
PT
3.1. Spawning of Siganus sutor
ED
M
and hormone administration, using the PRIMER software v6 (Clarke and Gorley, 2006).
CE
About 33.3 ± 18.2, 46.7 ± 20.0, 80.0 ± 13.3, 73.3 ± 19.4, 33.3 ± 18.2 and 26.7 ± 12.4 % of injected fishes spawned on May 1, 8, 15 and 22 and June 1 and 8, respectively. A significantly
AC
greater number of spawned fish (70.83 ± 16 %) was observed in treatment L2 (20 μg/kg LHRHa2) compared with the 16.7 ± 10.9, 45.8 ± 14, 45.8 ± 16 and 4.17 ± 4.17 %, which was observed in treatments L1, H, HM and C, respectively (Table 1). 3.2. Egg production and hatching rate of eggs The egg numbers per injected females on different days were 223200 ± 84800, 237666 ± 95288, 537733 ± 80412, 464800 ± 82180, 213400 ± 85447, and 199933 ± 88859, which were
ACCEPTED MANUSCRIPT obtained on May 1, 8, 15 and 22 and June1 and 8, respectively. A significant difference was observed between egg numbers per injected fish treated on May 15 and the other days (Fig. 1). Females in treatment L2 produced a significantly greater number of eggs per injected female (469083 ± 70453 eggs) in comparison with females in treatments L1, H, HM and C, which
T
produced 103375 ± 48872, 294500 ± 74348, 287458 ± 72856, and 18542 ± 18000 eggs (per
IP
injected fish), respectively (p < 0.05) (Fig. 2).
(
)
=
+ 13.119BW –
US
of S. sutor and the regression equation was calculated as
CR
A polynomial relationship was observed between relative fecundity and total body weight
2312.1 (R² = 0.55 and n=42) (Fig. 3).
AN
Hatching rate of 86.25 ± 1.38, 85.05 ± 0.82, 86.7 ± 0.91, 84.9 ± 0.51 and 85 % were
M
observed in treatments L1, L2, H, HM and C, respectively and there was no significant difference
ED
among hatching rates of the various treatments.
PT
3.3. Appropriate time and hormonal treatment for induced spawning in Siganus sutor According to cluster analysis of egg production of Siganus sutor , the best time for induced
CE
spawning of S. sutor was between 15th to 22nd May (7th to 14th lunar period) and the best
and 5)
AC
hormone for induced spawning was LHRHa2 at a dosage of 20 µg/kg BW of females (Figures 4
4. Discussion
Results of this study indicated the successful induced spawning of S. sutor using injection of different dosages of LHRHa2, HCG and HCG combined with metoclopramide. Similar results were obtained for induced spawning of rabbitfishes S. argenteus, S. revulatus and S. luridus
ACCEPTED MANUSCRIPT using 500 IU HCG/kg body weight of females (Popper et al., 1979). Induced spawning of S. guttatus occurred using injection of 500 IU/fish or about 2 IU/g body weight (Juario et al.,
1985). In another study spontaneous spawning and natural fertilization of the eggs occurred following ovulation in the fish injected with 500 IU HCG between 17 and 18 hours after the last
IP
T
of a series of injections delivered at 24-hour intervals to S. vermiculatus (Avila, 1984).
CR
The number of spawned fish and eggs production of S. sutor significantly varied on various days of the reproductive season (Table 1). All fishes in treatments L2, H and HM on days
US
15 and 22 May responded to these hormones and successfully spawned two or three days after
AN
the second injection of hormones. Only one fish out of 24 in treatment C (1 ml of saline water 0.9%) spawned on 15 May and produced a small number of eggs. This result indicated that S.
M
sutor does not spawn in captivity without using any hormonal treatment. Similarly, injection of
ED
LHRHa at dosages of 25, 50 and 100 μg/kg led to spawning in sand bass (Paralabrax maculatofasciatus) after the second injection of LHRHa applied to females in all treated groups,
PT
and no females injected with saline solution spawned (Alcántar-Vázquez et al., 2016). Almost all
CE
of fishes which are important for commercial aquaculture industry exhibit reproductive dysfunctions that probably result from the combination of captivity-induced stress and the lack
AC
of the appropriate environmental conditions for spawning, thus, females cannot undergo final oocyte maturation, and thus ovulation and spawning don’t occur (Zohar and Mylonas, 2001). In this study the maximum number of injected females spawned on days 17, 18, 24 and 25 of May 2016, which was during the 9th - 17th days of the lunar month. It has been reported that growth, feeding, migration and reproduction behavior of many reef fishes may be affected by the changes in lunar phases (Boujard, 1992; Takemura et al., 2004; Horký et al., 2006).
ACCEPTED MANUSCRIPT All the fishes in treatment L2 which were treated on May 1, 8, 15 and 22 and June 1 and 8(days: 3, 22, 29, 7, 14, 22, 1 and 8, of lunar month), spawned successfully; these results showed that there was no relationship between spawning days of the fishes in treatment L2 and the lunar rhythm. None of the fishes responded to different hormonal treatments on April 12 or June15
T
(Table 1). Previous research about reproductive biology of S. sutor in Iran showed that there is
IP
one peak period of spawning from May to July (Fourooghifard et al., 2009). Therefore, it could
CR
be said, these days (April 12 and June 15) are out of the reproductive time cycle. In contrast, spawning of S. guttatus occurred throughout the year, simply by introducing one female with l-2
US
males into a spawning tank on the day of the first quarter moon (Hara et al., 1986). In some
AN
siganids such as S. doliatus, the ovary develops twice, from February to May and from August to
M
September (Park et al. 2006).
Cluster analysis of average eggs production showed a similarity between the effect of
ED
HCG and HCG combined with the dopamine antagonist (metoclopramide) on spawning of S.
PT
sutor (Fig. 5). Similarly, the results of injection of dopamine antagonists (chlorpromazine) on inducing spawning in Caspian Kutum, Rutilus frisii kutum showed that blocking effect of
CE
dopamine on gonadotropin hormone (GtH) secretion in Kutum is of less importance than in other
AC
cyprinids (Khara and Oryan, 2012). Inhibition of dopamine on maturity varies among different species in many fish families such as cyprinids and the African catfish, Clarias gariepinus (Mylonas and Zohar 2001). The maximum and minimum numbers of eggs (0.97 million and 0.35 million) were produced by a 623-g and a 380-g BW female, respectively. In other research, a 400-g captive S. guttatus broodstock had 0.8 million eggs, and a 520-g female produced 1.2 million eggs (Soletchnik, 1984). A 400-450-g female of S. rivulatus can produce about 1.05-1.22 million
ACCEPTED MANUSCRIPT eggs, and 400-450-g females of S. luridus produced about 0.838-0.986 million eggs, respectively (Bariche et al., 2009). The relationship between relative fecundity and total body weight (
(
)
=
+ 13.119BW - 2312.1) of S. sutor , showed that although the bigger females produce more eggs,
IP
T
the relative fecundity in a big fish (585 g BW;1661000 eggs kg/BW) is less than a smaller one
CR
(518 g BW; 1667000 eggs kg/BW). It can be concluded that in S. sutor, the medium females (450-550 g BW) are more suitable than the other ones for induced reproduction in captivity (Fig.
US
3). Cluster analysis showed that the best time for induced spawning of S. sutor was May 15 to 22 (however, this is maybe not entirely general and would change year to year according to the
AN
differences in temperature). The best hormone for induced spawning was LHRHa2 at a dosage of
M
20 µg/kg BW of females (Figs. 4 and 5). Previous research about reproductive biology of the shoemaker spinefoot rabbitfish, S. sutor, in culture conditions in Iran, showed that there is one
ED
peak period of spawning from May to July (Fourooghifard et al., 2009). According to the results
PT
of this study and the previous study, it could be concluded that the reproduction period of S.
AC
5. Conclusion
CE
sutor is fairly short in Iran and lasts from early May to mid-June.
The best result for Induced spawning of the shoemaker rabbit fish S. sutor can be achieved using
at a dosage of 20 µg/kg BW of females. According to the results from this study
and previous research (Fourooghifard et al., 2009), the reproduction period of S. sutor is fairly short in Iran and lasts from early May to mid-June. The females with body weight of 450-550 g are most suitable for induced reproduction in captivity and produce the maximum eggs. 6. Acknowledgments
ACCEPTED MANUSCRIPT The authors are grateful for the project support provided by the Director of Persian Gulf and Oman Sea Ecological Research Institute (PGOSERI), Bandar Abbass, Iran. References
T
Agembe, S., 2012. Estimation of important reproductive parameters for management of the Shoemaker
IP
spinefoot rabbitfish (Siganus sutor) in southern Kenya. International Journal of Marine Science 2(4),
CR
24-30.
Alcántar-Vázquez, J.P., Pliego-Cortés, H.S., Dumas, S., Peña-Martínez, R., Rosales-Velázquez, M.,
US
Pintos-Terán, P., 2016. Effects of a luteinizing hormone-releasing hormone analogue (LHRHa) on the reproductive performance of spotted sand bass Paralabrax maculatofasciatus (Percoidei:
AN
Serranidae). Latin American Journal of Aquatic Research 44(3), 487-496. Ashraf, M.A., Akar, A., 2013. Spawning induction in silver carp (Hypothalamichthys molitrix) using carp
M
pituitary extract (CPE) alone, receptal (Buserelin acetate) and cystorelin (GnRh) with or without
ED
Dopamine antagonists In Proceedings of the 6th Global Fisheries and Aquaculture Research Conference, Hurghada, Egypt, 27-30 September 2013, 137-148.
PT
Avila, E.M., 1984. Hormone-induced spawning and embryonic development of the rabbitfish, Siganus
CE
vermiculatus (Pisces: Siganidae). Philippine Journal of Science 21, 75-108. Ayson, F.G., 1989. The effect of stress on spawning of brood fish and survival of larvae of the rabbitfish,
AC
Siganus guttatus (Bloch). Aquaculture 80, 241-246. Ayson, F.G., 1991. Induced spawning of rabbitfish, Siganus guttatus(Bloch) using human chorionic gonadotropin (HCG). Aquaculture 95, 133-137. Bariche, M., Sadek, R., Azzurro, E., 2009. Fecundity and condition of successful invaders: Siganus rivulatus and S. luridus (Actinopterygii: Perciformes: Siganidae) in the Eastern Mediterranean Sea. Acta Ichthyologica et Piscatoria 39(1), 11-18. Boujard, T., 1992. Space-time organization of riverine fish communities in French Guiana. Environmental Biology of Fishes 34, 235-246.
ACCEPTED MANUSCRIPT Bryan, P.G., Madraisau, B.B., 1977. Larval rearing and development of Siganus lineatus (Pisces: Siganidae) from hatching through metamorphosis. Aquaculture, 10, 243-252. Clarke, K.R., Gorley, R., 2006. PRIMER v6 1. 10: User Manual/Tutorial (Plymouth Routines in Multivariate Ecological Research). Plymouth, UK., 192 p.
T
Duarte, C.M., Marbá, N., Holmer, M., 2007. Rapid domestication of marine species. Science
IP
316(5823), 382-383.
CR
Duray, M.N., Juario, J.V., 1988. Broodstock management and seed production of the rabbitfish Siganus guttatus (Bloch) and the sea bass Lates calcarifer (Bloch), In: Juario, J.V., Benitez, L.V., (editors),
US
Proceedings of the Seminar on Aquaculture Development in Southeast Asia, 8-12 September 1987, Iloilo City, Philippines, SEAFDEC Aquaculture Department 195-210.
AN
Foroughifard, H., Tazikeh, E., Gharavi, B., Zarshenas, G.A., 2007. Artificial Reproduction Of Siganus Sutor Using Lhrha2 And HCG In Hormozgan Province, South Iran. Iranian Scientific Fisheries
M
Journal 16 (1), 161-168.
ED
Fourooghifard, H., Daghooghi, B., Aftabsavar, Y., 2009. Reproductive biology of the white spotted rabbit fish, Siganus sutor, in culture conditions (in persian). Iranian Scientific Fisheries Journal 18, 119-
PT
128.
CE
Fourooghifard, H., Matinfar, A., Abdolalian, E., Moezzi, M., Roohani, G.K., Kamali, E., Allen, S., Zahedi, M., 2017. Egg production and larval rearing of orange-spotted grouper (Epinephelus
AC
coioides) using reared broodstocks in Hormozgan Province, Iran. Iranian Journal of Fisheries Sciences 16 (3), 984-992. Hara, S., Duray, M.N., Parazo, M., Taki, Y., 1986. Year-round spawning and seed production of the rabbitfish, Siganus guttatus. Aquaculture 59, 259-272. Horký, P., Slavík, O., Bartos, L., Kolárová, J., Randák, T., 2006. The effect of the moon phase and seasonality on the behaviour of pikeperch in the Elbe River. Folia Zoologica 55, 411-417
ACCEPTED MANUSCRIPT Islam, M., Sultana, N., Hossain, M.B., Mondal, S., 2012. Estimation of fecundity and gonadosomatic index (GSI) of gangetic whiting, Sillaginopsis panijus (Hamilton, 1822) from the Meghna River Estuary, Bangladesh. World Applied Sciences Journal 17, 1253-1260. Juario, J.V., Duray, M.N., Duray, V.M., Nacario, J.F., Almendras, J.M., 1985. Breeding and larval rearing
T
of the rabbitfish, Siganus guttatus (Bloch). Aquaculture 44, 91-101.
IP
Kamukuru, A.T., 2006. Reproductive Biology of the White Spotted Rabbitfish, Siganus Sutor (Pisces:
CR
Siganidae) from Basket Trap Fishery in Dar Es Salaam Marine Reserves Systems, WIOMSA Tanzania, 60 p.
US
Khara, H., Oryan, S., 2012. Effects of LHRH-A2 and chlorpromazine (dopamine antagonists) on inducing spawning in Caspian Kutum, Rutilus frisii kutum, from the southwest of the Caspian Sea. Caspian
AN
Journal of Environmental Sciences 10, 33-42.
Kucharczyk, D., Kujawa, R., Mamcarz, A., Targońska-Dietrich, K., Wyszomirska, E., Glogowski, J.,
M
Babiak, I., Szabo, T., 2005. Induced spawning in bream (Abramis brama L.) using pellets containing
ED
GnRH. Czech Journal of Animal Science 50, 89-95. Lam, T., 1974. Siganids: their biology and mariculture potential. Aquaculture 3, 325-354.
PT
Lam, T., Soh, C., 1975. Effect of photoperiod on gonadal maturation in the rabbitfish, Siganus
CE
canaliculatus Park 1797. Aquaculture 5, 407-410. Marte, C., 1989. Hormone-induced spawning of cultured tropical finfishes. In Advances in Tropical
AC
Aquaculture, Workshop at Tahiti, French Polynesia, 20 Feb-4 Mar 1989, IFREMER, Actes de Colloque 9, 519- 539. Mylonas, C.C., Fostier, A., Zanuy, S., 2010. Broodstock management and hormonal manipulations of fish reproduction. General and Comparative Endocrinology 165, 516-534. Naeem, M., Zuberi, A., Salam, A., Ashraf, M., Elahi, N., Ali, M., Ishtiaq, A., Malik, T., Khan, M.J., Ayaz, M.M., 2011. Induced spawning, fecundity, fertilization rate and hatching rate of Grass carp (Ctenopharyngodon idella) by using a single intramuscular injection of ovaprimC at a fish hatchery in Faisalabad, Pakistan. African Journal of Biotechnology, 10, 11048-11053.
ACCEPTED MANUSCRIPT Nelson, S., G., Lock, S.A., Collins, L.A., 1992. Growth of the Rabbitfish Siganus Randalli Woodland in Relation to the Feasibility of Its Culture on Guam. University of Guam Marine Laboratory. Technical Report No. 97, Micronesia, 30p. Park Y.J., Takemura, A, Lee, Y.D., 2006. Lunar-synchronized reproductive activity in the pencil-streaked
T
rabbitfish Siganus doliatus in the Chuuk Lagoon, Micronesia. Ichthyological Research 53, 179-181.
IP
Popper, D., Gordin, H., Kissil, G.W., 1973. Fertilization and hatching of rabbitfish Siganus rivulatus.
CR
Aquaculture 2, 37-44.
Popper, D., May, R., Lichatowich, T., 1976. An experiment in rearing larval Siganus vermiculatus
US
(Valenciennes) and some observations on its spawning cycle. Aquaculture 7, 281-290. Popper, D., Pitt, R., Zohar, Y., 1979. Experiments on the propagation of Red Sea siganids and some
AN
notes on their reproduction in nature. Aquaculture 16, 177-181.
Rahman, M., Takemura, A., Park, Y., Takano, K., 2003. Lunar cycle in the reproductive activity in the
M
forktail rabbitfish. Fish Physiology and Biochemistry 28, 443-444.
ED
Rahman, M.S., Takemura, A., Takano, K., 2001. Lunar synchronization of testicular development and steroidogenesis in rabbitfish. Comparative Biochemistry and Physiology Part B: Biochemistry and
PT
Molecular Biology 129, 367-373.
CE
Randall, J.E., Allen, G.R., Steene, R.C., 1997. Fishes of the Great Barrier Reef and Coral Sea. University of Hawaii Press, Honolulu, Hawaii , 608 p.
AC
Soletchnik, P., 1984. Aspects of nutrition and reproduction in Siganus guttatus with emphasis on applications to aquaculture. Southeast Asian Fisheries Development Center, Iloilo, philippines. Takemura, A ,.Rahman, M., Nakamura, S., Park, Y.J., Takano, K., 2004. Lunar cycles and reproductive activity in reef fishes with particular attention to rabbitfishes. Fish and Fisheries 5, 317-328. Takemura, A., Rahman, M., Park, Y., 2010. External and internal controls of lunar-related reproductive rhythms in fishes. Journal of Fish Biology 76, 7-26.
ACCEPTED MANUSCRIPT Zohar, Y., Mylonas, C.C., 2001. Endocrine manipulations of spawning in cultured fish: from hormones to genes, In: Lee , C. S. (Edtor), Reproductive Biotechnology in Finfish Aquaculture. Elsvier, Netherlands, pp. 99-136.
T
Fig. 1 Egg production of Siganus. sutor on various days of reproduction season (the numbers in
IP
parentheses show the lunar days) (Mean ±Se).
CR
Fig. 2 Egg production of S. sutor treated by different hormone administrations (Mean ±Se).
US
Fig. 3 The regression between relative fecundity and total body weight of S. sutor treated by
AN
hormone administration.
Fig. 4 Dendrogram of hierarchical analysis of average egg production in S. sutor treated by
M
different hormone administrations during the reproduction season.
ED
Fig. 5. Dendrogram of hierarchical analysis of average egg production in S. sutor treated by
PT
different hormone administrations.
Date of hormone injection May 8 (29)
May 15 (7)
May 22 (14)
June 1 (22)
June 8 (1)
June 15 (8)
Mean ± SE
0
0
0
66.67
66.67
0
0
0
16.7±10.9
0
100
100
100
100
100
66.7
0
70.8±16
H
0
33.3
66.7
100
100
33.3
33.3
0
45.8±14
HM
0
33.3
66.7
100
100
33.3
33.3
0
45.8±14
C
0
0
0
33.3
0
0
0
0
4.2±4.2
Mean ± SE
0
33.3±18.3
46.7±2
80±13.3
73.3±19.4
33.3±18.3
26.7±12.5
0
L1 L2
AC
May 1 (22)
Treatment
April 12 (3)
CE
Table 1 Total spawned females (%) of S. sutor treated by all types of hormone administration on various days of reproductive season (the numbers in parentheses show the lunar days) (Mean ± Se).
ACCEPTED MANUSCRIPT L1= LHRHa2 hormone at dosage of 10 μg/Kg body weight (BW) ; L2 = LHRHa2 hormone at dosage of 20 μg/Kg BW ; H= HCG hormone at a dosage of 2000 IU/kg BW ; HM = HCG hormone at a dosage of 2000 IU/kg BW combined with 5 mg metoclopramide/Kg BW ; C = 1 ml 0.9% saline water.
Highlights: The information of the present study can be helpful to fish breeders in planning for the most efficient egg production and rearing of this important species.
T
IP
A significantly greater number of spawned fish (70.83 ± 9.48 %), was observed in treatment L2
CR
US AN M ED PT CE AC
compared with the other treatments (L1: 16.67±7.77 %, H: 41.67± 10.28 %, HM: 41.67±10.28 % and C: 4.17± 4.17 %). Cluster analysis showed that the best time for induced spawning of S. sutor was 15 to 22 May and the best hormone for induced spawning was LHRHa2 at a dosage of 20µg/kg BW of females.
Hojjatollah Fourooghifard, Kiuomars Roohani Ghadikolaee, Samuel Allen, Maryam Moezzi, Reza Dehghani, Mohammad Reza Zahedi PII: DOI: Reference:
S0044-8486(19)30164-4 https://doi.org/10.1016/j.aquaculture.2019.05.062 AQUA 634177
To appear in:
aquaculture
Received date: Revised date: Accepted date:
21 January 2019 26 May 2019 26 May 2019
Please cite this article as: H. Fourooghifard, K.R. Ghadikolaee, S. Allen, et al., Determining the appropriate time and hormone administration for induced spawning and egg production of the shoemaker spinefoot rabbitfish, Siganus sutor (Valenciennes, 1835) during the reproductive season, aquaculture, https://doi.org/10.1016/ j.aquaculture.2019.05.062
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT Determining the appropriate time and hormone administration for induced spawning and egg production of the shoemaker spinefoot rabbitfish, Siganus sutor (Valenciennes, 1835) during the reproductive season Hojjatollah Fourooghifarda,* [email protected], Kiuomars Roohani Ghadikolaeea
T
[email protected], Samuel Allenb [email protected], Maryam Moezzia
Persian Gulf and Oman Sea Ecological Research Center, Iranian Fisheries Sciences Research
US
a
CR
Reza Zahedia [email protected]
IP
[email protected], Reza Dehghania [email protected], Mohammad
AN
Institute, Agricultural Research, Education and Extension Organization(AREEO), Bandar Abbas, Iran.
Plant and Environmental Sciences Department, New Mexico State University Agricultural Science
ED
Center, Farmington, New Mexico, USA
M
b
*Corresponding author at: Persian Gulf and Oman Sea Ecological Research Center, Iranian
PT
Fisheries Sciences Research Institute, Agricultural Research, Education and Extension
AC
Abstract
CE
Organization (AREEO), Bandar Abbas, Iran
This study was conducted to determine the appropriate time and hormone administration protocol for induced spawning of the shoemaker spinefoot rabbitfish, Siganus sutor. Four different hormone administration treatments including LHRHa2 hormone at dosages of 10 and 20 μg/kg body weight (BW) (treatment
and
), HCG hormone alone at a dosage of 2000 IU/kg
BW (treatment H) and combined with 5 mg metoclopramide/kg BW (treatment HM) plus the control, 1 ml 0.9% saline water (treatment C) were administered on eight dates in mid-2016
ACCEPTED MANUSCRIPT (April 12, May 1, 8, 15 and 22 and June 1,8 and 15). Females (470.14 ± 8.56 g BW) received the hormone injection twice over a 24-hour interval and all males received HCG hormone at a dosage of 2000 IU/kg BW; hence, each fish was used just once for the injection of hormone. A significantly greater number of spawned fish (70.8 ± 16 %) was observed in treatment L2
T
compared with the other treatments (L1: 16.7 ± 10.8 %; H: 45.8 ± 14 %; HM: 45.8 ± 14 %; and
IP
C: 4.2 ± 0.2 %). Significantly greater numbers of eggs per female (469083 ± 70453 eggs) were
CR
obtained in treatment L2 compared with the other treatments (L1: 103375 ± 48872 eggs; H:
US
294500 ± 74348 eggs; HM: 287458 ± 72856 eggs; and C: 18542 ± 18000 eggs). Cluster analysis showed that the best time for induced spawning of S. sutor was 15 to 22 May and the best
AN
hormone for induced spawning was LHRHa2 at a dosage of 20 µg/kg BW of females. Such information can be helpful in the management of breeding programs for this important fish
M
species.
ED
Keywords:
CE
1. Introduction
PT
Rabbitfish; Siganus sutor; HCG hormone; LHRHa2 hormone; Induced spawning.
AC
Development of commercial aquaculture has led to an increase in the number of aquatic species under domestication efforts (Duarte et al., 2007). One of the important prerequisites for domestication and improvement of a sustainable aquaculture industry is the ability to control the reproductive processes and to obtain high quality of eggs and sperm of fish in captivity (Mylonas et al., 2010). Siganids are generally regarded as good food fishes in spite of their relatively small size, and some species have been cultured because of their herbivorous food habits, rapid growth and economic value (Randall et al.,1997). Artificial reproduction of some species of rabbitfishes
ACCEPTED MANUSCRIPT has been investigated in Siganus gattatus, S. revulatus, S. vemiculatus, S. lineatus and S. canaliculatus from the 1970s (Ayson, 1991; Bryan and Madraisau, 1977; Popper et al., 1976; Lam and Soh, 1975; Popper et al., 1973). There are a few reports about the reproduction biology of this species from various parts of the world (Agembe, 2012; Kamukuru, 2006). Some species
T
of rabbitfish such as S. gattatus can spawn without hormonal treatment, every month throughout
IP
the year (Hara et al., 1986). Induced spawning of rabbitfish has been successfully carried out
CR
using HCG injection (500 IU/ fish or about 2 IU/g body weight) in S. gattatus and S. canaliculatus (Juario et al., 1985; Lam, 1974). Effect of using metoclopramide as a dopamine
US
antagonist has been used to facilitate the induction spawning of some species such as silver carp
AN
(Hypothalamichthys molitrix), bream (Abramis brama L.) and common carp (Cyprnus carpio)(Ashraf and Akar, 2013; Kucharczyk et al., 2005; ). It has been reported that spawning
M
behavior of rabbitfish can be related to lunar periodicity (Rahman et al., 2003; Rahman et al.,
ED
2001; Hara et al., 1986). Some species such as white-spotted spinefoot Siganus canaliculatus, spawn around the new moon from April to June, little spinefoot S. spinus spawn around the new
PT
moon from May to July, the goldlined spinefoot S. guttatus, and the streamlined spinefoot S.
CE
argenteus, spawn around the first quarter moon from June to July (Takemura et al., 2010). A first study about the induced spawning of S. sutor was conducted in Iran in 2005 (Foroughifard et al.,
AC
2007). Although the feasibility of broodstocking and maturation in rabbit fish, was studied by Fourooghifard et al. (2009), there were many unknowns to achieve the acceptable result for induced spawning of this species. Hence, a new study was conducted to determine the appropriate time and hormonal treatment for induced spawning and egg production of this species using LHRHa2 and HCG hormones with or without metoclopramide as a dopamine antagonist.
ACCEPTED MANUSCRIPT 2. Materials and methods 2.1. Design of experiment The experiments were conducted at the Persian Gulf and Oman Sea Ecological Research Institute (PGOSERI), Bandar Abbas, Iran, from March to June 2016. Four different hormonal
T
treatments including LHRHa2 hormone at a dosage of 10 μg/ kg body weight (Treatment L1),
CR
IP
LHRHa2 hormone at a dosage of 20 μg/ kg BW (Treatment L2), HCG hormone at a dosage of 2000 IU/Kg BW alone (treatment H) and combined with 5 mg metoclopramide/kg BW
US
(treatment HM), and a control treatment with 1 ml 0.9% saline solution without any hormone (Treatment C) were performed to survey their effects on induced spawning of Siganus sutor at
AN
the environmental temperature of ~26–30 °C. The LHRHa2 hormone was provided from Ningbo
M
Sansheng Pharmaceutical Co., China and the HCG and metoclopramide were provided from Daru Pakhsh Pharmaceutical Co., Iran. Injections were carried out in mid-2016 on eight different
ED
days ( April 12; May 1, 8, 15 and 22; June 1, 8 and 15). Each treatment had three replications
PT
(one female and one male for each replication), therefore 15 females and 15 males were treated on each day (for a total of 120 females and 120 males during the reproduction season). Each fish
CE
was used just once for the injection of hormone during the experiments. Each pair of injected
AC
fish (1 female and 1 male) was introduced to a 1-m3 separate spawning tank. Since , like other Siganids, S. sutor eggs are adhesive, egg collectors consisting of corrugated fiberglass sheets were placed at the bottom of spawning tanks prior to spawning (Nelson et al., 1992). All the experiments were conducted under natural light period and temperature. 2.2. Broodstocks of Siganus sutor A total of 400 adults of Siganus sutor (387.46 ± 11.73 g body weight) (mean± Se) were collected from the coastal waters of Lavan Island, Iran, and were stocked in 5-m3 fiberglass tanks
ACCEPTED MANUSCRIPT at a density of 8 fish/m3 (3-4 kg/m3) (Duray and Juario, 1988). The fish were reared until their average body weight reached 470.14 ± 8.56 g at the reproduction season. Fishes were fed twice a day with commercial shrimp pellets (40% protein, 5% fat) and crab meat at a rate of 3% body weight (Fourooghifard et al., 2009).
IP
T
2.3. Injection of hormones
CR
The LHRHa2 and HCG powder were diluted in 1 ml 0.9% saline solution just prior to injection. In the treatment HM, the HCG powder was diluted in 1 ml 0.9% saline solution and
US
mixed with 0.5 ml of 10 mg /1ml metoclopramide injection vial. Female fish were distinguished from the males by their swollen and soft abdomens and males were identified by the flowing of
AN
milt through the genital pore after a gentle pressure on their abdomen (Hara et al., 1986).
M
Hormones were injected intramuscularly below the dorsal fin, twice (by 24 hour intervals)
ED
to each female at equal doses, between 8-9 a.m., during April–May, when the environmental temperature was about 26-28 °C (Foroughifard et al., 2007; Marte, 1989). All males were
PT
injected using HCG hormone at a dosage of 2000 IU/kg BW below the dorsal fin at the same
CE
time as the second injections of females (Duray and Juario, 1988). 2.4. Fecundity and hatching rate of eggs
AC
After the spawning of fishes, the egg collectors were removed from the spawning tanks and all eggs were washed and collected onto a sieve and were weighted. Three samples of egg (0.4-0.5 g) were taken from the sieve and the eggs of each sample were counted and the average was obtained. The following formula was used to calculate the Fecundity (Islam et al., 2012). Fecundity (F) = n × G/g, where “n” is the number of eggs in each sample, “G” is the weight of total eggs collected on the sieve, and “g” is the weight of sample
ACCEPTED MANUSCRIPT A batch of 5000 eggs belonging to each female was stocked in a 300-liter round polyethylene tank at a density of 25 eggs per liter to calculate the hatching rate of eggs (Fourooghifard et al., 2017). Hatching rates were obtained by counting the larvae at 1 DAH and using the following formula (Naeem et al., 2011): Hatching rate = 100(Number of larvae/ Total
IP
T
eggs).
CR
2.5. Data analysis
Data were analyzed using SPSS 22.0 (IBM Corp., Armonk, NY, USA) software through
US
the application of parametric tests. The significance of differences was tested by one-way
AN
analysis of variance (ANOVA) and the post-hoc Duncan’s test. Differences were considered significant at p < 0.05. A hierarchical analysis was used for determination of the appropriate time
3. Results
PT
3.1. Spawning of Siganus sutor
ED
M
and hormone administration, using the PRIMER software v6 (Clarke and Gorley, 2006).
CE
About 33.3 ± 18.2, 46.7 ± 20.0, 80.0 ± 13.3, 73.3 ± 19.4, 33.3 ± 18.2 and 26.7 ± 12.4 % of injected fishes spawned on May 1, 8, 15 and 22 and June 1 and 8, respectively. A significantly
AC
greater number of spawned fish (70.83 ± 16 %) was observed in treatment L2 (20 μg/kg LHRHa2) compared with the 16.7 ± 10.9, 45.8 ± 14, 45.8 ± 16 and 4.17 ± 4.17 %, which was observed in treatments L1, H, HM and C, respectively (Table 1). 3.2. Egg production and hatching rate of eggs The egg numbers per injected females on different days were 223200 ± 84800, 237666 ± 95288, 537733 ± 80412, 464800 ± 82180, 213400 ± 85447, and 199933 ± 88859, which were
ACCEPTED MANUSCRIPT obtained on May 1, 8, 15 and 22 and June1 and 8, respectively. A significant difference was observed between egg numbers per injected fish treated on May 15 and the other days (Fig. 1). Females in treatment L2 produced a significantly greater number of eggs per injected female (469083 ± 70453 eggs) in comparison with females in treatments L1, H, HM and C, which
T
produced 103375 ± 48872, 294500 ± 74348, 287458 ± 72856, and 18542 ± 18000 eggs (per
IP
injected fish), respectively (p < 0.05) (Fig. 2).
(
)
=
+ 13.119BW –
US
of S. sutor and the regression equation was calculated as
CR
A polynomial relationship was observed between relative fecundity and total body weight
2312.1 (R² = 0.55 and n=42) (Fig. 3).
AN
Hatching rate of 86.25 ± 1.38, 85.05 ± 0.82, 86.7 ± 0.91, 84.9 ± 0.51 and 85 % were
M
observed in treatments L1, L2, H, HM and C, respectively and there was no significant difference
ED
among hatching rates of the various treatments.
PT
3.3. Appropriate time and hormonal treatment for induced spawning in Siganus sutor According to cluster analysis of egg production of Siganus sutor , the best time for induced
CE
spawning of S. sutor was between 15th to 22nd May (7th to 14th lunar period) and the best
and 5)
AC
hormone for induced spawning was LHRHa2 at a dosage of 20 µg/kg BW of females (Figures 4
4. Discussion
Results of this study indicated the successful induced spawning of S. sutor using injection of different dosages of LHRHa2, HCG and HCG combined with metoclopramide. Similar results were obtained for induced spawning of rabbitfishes S. argenteus, S. revulatus and S. luridus
ACCEPTED MANUSCRIPT using 500 IU HCG/kg body weight of females (Popper et al., 1979). Induced spawning of S. guttatus occurred using injection of 500 IU/fish or about 2 IU/g body weight (Juario et al.,
1985). In another study spontaneous spawning and natural fertilization of the eggs occurred following ovulation in the fish injected with 500 IU HCG between 17 and 18 hours after the last
IP
T
of a series of injections delivered at 24-hour intervals to S. vermiculatus (Avila, 1984).
CR
The number of spawned fish and eggs production of S. sutor significantly varied on various days of the reproductive season (Table 1). All fishes in treatments L2, H and HM on days
US
15 and 22 May responded to these hormones and successfully spawned two or three days after
AN
the second injection of hormones. Only one fish out of 24 in treatment C (1 ml of saline water 0.9%) spawned on 15 May and produced a small number of eggs. This result indicated that S.
M
sutor does not spawn in captivity without using any hormonal treatment. Similarly, injection of
ED
LHRHa at dosages of 25, 50 and 100 μg/kg led to spawning in sand bass (Paralabrax maculatofasciatus) after the second injection of LHRHa applied to females in all treated groups,
PT
and no females injected with saline solution spawned (Alcántar-Vázquez et al., 2016). Almost all
CE
of fishes which are important for commercial aquaculture industry exhibit reproductive dysfunctions that probably result from the combination of captivity-induced stress and the lack
AC
of the appropriate environmental conditions for spawning, thus, females cannot undergo final oocyte maturation, and thus ovulation and spawning don’t occur (Zohar and Mylonas, 2001). In this study the maximum number of injected females spawned on days 17, 18, 24 and 25 of May 2016, which was during the 9th - 17th days of the lunar month. It has been reported that growth, feeding, migration and reproduction behavior of many reef fishes may be affected by the changes in lunar phases (Boujard, 1992; Takemura et al., 2004; Horký et al., 2006).
ACCEPTED MANUSCRIPT All the fishes in treatment L2 which were treated on May 1, 8, 15 and 22 and June 1 and 8(days: 3, 22, 29, 7, 14, 22, 1 and 8, of lunar month), spawned successfully; these results showed that there was no relationship between spawning days of the fishes in treatment L2 and the lunar rhythm. None of the fishes responded to different hormonal treatments on April 12 or June15
T
(Table 1). Previous research about reproductive biology of S. sutor in Iran showed that there is
IP
one peak period of spawning from May to July (Fourooghifard et al., 2009). Therefore, it could
CR
be said, these days (April 12 and June 15) are out of the reproductive time cycle. In contrast, spawning of S. guttatus occurred throughout the year, simply by introducing one female with l-2
US
males into a spawning tank on the day of the first quarter moon (Hara et al., 1986). In some
AN
siganids such as S. doliatus, the ovary develops twice, from February to May and from August to
M
September (Park et al. 2006).
Cluster analysis of average eggs production showed a similarity between the effect of
ED
HCG and HCG combined with the dopamine antagonist (metoclopramide) on spawning of S.
PT
sutor (Fig. 5). Similarly, the results of injection of dopamine antagonists (chlorpromazine) on inducing spawning in Caspian Kutum, Rutilus frisii kutum showed that blocking effect of
CE
dopamine on gonadotropin hormone (GtH) secretion in Kutum is of less importance than in other
AC
cyprinids (Khara and Oryan, 2012). Inhibition of dopamine on maturity varies among different species in many fish families such as cyprinids and the African catfish, Clarias gariepinus (Mylonas and Zohar 2001). The maximum and minimum numbers of eggs (0.97 million and 0.35 million) were produced by a 623-g and a 380-g BW female, respectively. In other research, a 400-g captive S. guttatus broodstock had 0.8 million eggs, and a 520-g female produced 1.2 million eggs (Soletchnik, 1984). A 400-450-g female of S. rivulatus can produce about 1.05-1.22 million
ACCEPTED MANUSCRIPT eggs, and 400-450-g females of S. luridus produced about 0.838-0.986 million eggs, respectively (Bariche et al., 2009). The relationship between relative fecundity and total body weight (
(
)
=
+ 13.119BW - 2312.1) of S. sutor , showed that although the bigger females produce more eggs,
IP
T
the relative fecundity in a big fish (585 g BW;1661000 eggs kg/BW) is less than a smaller one
CR
(518 g BW; 1667000 eggs kg/BW). It can be concluded that in S. sutor, the medium females (450-550 g BW) are more suitable than the other ones for induced reproduction in captivity (Fig.
US
3). Cluster analysis showed that the best time for induced spawning of S. sutor was May 15 to 22 (however, this is maybe not entirely general and would change year to year according to the
AN
differences in temperature). The best hormone for induced spawning was LHRHa2 at a dosage of
M
20 µg/kg BW of females (Figs. 4 and 5). Previous research about reproductive biology of the shoemaker spinefoot rabbitfish, S. sutor, in culture conditions in Iran, showed that there is one
ED
peak period of spawning from May to July (Fourooghifard et al., 2009). According to the results
PT
of this study and the previous study, it could be concluded that the reproduction period of S.
AC
5. Conclusion
CE
sutor is fairly short in Iran and lasts from early May to mid-June.
The best result for Induced spawning of the shoemaker rabbit fish S. sutor can be achieved using
at a dosage of 20 µg/kg BW of females. According to the results from this study
and previous research (Fourooghifard et al., 2009), the reproduction period of S. sutor is fairly short in Iran and lasts from early May to mid-June. The females with body weight of 450-550 g are most suitable for induced reproduction in captivity and produce the maximum eggs. 6. Acknowledgments
ACCEPTED MANUSCRIPT The authors are grateful for the project support provided by the Director of Persian Gulf and Oman Sea Ecological Research Institute (PGOSERI), Bandar Abbass, Iran. References
T
Agembe, S., 2012. Estimation of important reproductive parameters for management of the Shoemaker
IP
spinefoot rabbitfish (Siganus sutor) in southern Kenya. International Journal of Marine Science 2(4),
CR
24-30.
Alcántar-Vázquez, J.P., Pliego-Cortés, H.S., Dumas, S., Peña-Martínez, R., Rosales-Velázquez, M.,
US
Pintos-Terán, P., 2016. Effects of a luteinizing hormone-releasing hormone analogue (LHRHa) on the reproductive performance of spotted sand bass Paralabrax maculatofasciatus (Percoidei:
AN
Serranidae). Latin American Journal of Aquatic Research 44(3), 487-496. Ashraf, M.A., Akar, A., 2013. Spawning induction in silver carp (Hypothalamichthys molitrix) using carp
M
pituitary extract (CPE) alone, receptal (Buserelin acetate) and cystorelin (GnRh) with or without
ED
Dopamine antagonists In Proceedings of the 6th Global Fisheries and Aquaculture Research Conference, Hurghada, Egypt, 27-30 September 2013, 137-148.
PT
Avila, E.M., 1984. Hormone-induced spawning and embryonic development of the rabbitfish, Siganus
CE
vermiculatus (Pisces: Siganidae). Philippine Journal of Science 21, 75-108. Ayson, F.G., 1989. The effect of stress on spawning of brood fish and survival of larvae of the rabbitfish,
AC
Siganus guttatus (Bloch). Aquaculture 80, 241-246. Ayson, F.G., 1991. Induced spawning of rabbitfish, Siganus guttatus(Bloch) using human chorionic gonadotropin (HCG). Aquaculture 95, 133-137. Bariche, M., Sadek, R., Azzurro, E., 2009. Fecundity and condition of successful invaders: Siganus rivulatus and S. luridus (Actinopterygii: Perciformes: Siganidae) in the Eastern Mediterranean Sea. Acta Ichthyologica et Piscatoria 39(1), 11-18. Boujard, T., 1992. Space-time organization of riverine fish communities in French Guiana. Environmental Biology of Fishes 34, 235-246.
ACCEPTED MANUSCRIPT Bryan, P.G., Madraisau, B.B., 1977. Larval rearing and development of Siganus lineatus (Pisces: Siganidae) from hatching through metamorphosis. Aquaculture, 10, 243-252. Clarke, K.R., Gorley, R., 2006. PRIMER v6 1. 10: User Manual/Tutorial (Plymouth Routines in Multivariate Ecological Research). Plymouth, UK., 192 p.
T
Duarte, C.M., Marbá, N., Holmer, M., 2007. Rapid domestication of marine species. Science
IP
316(5823), 382-383.
CR
Duray, M.N., Juario, J.V., 1988. Broodstock management and seed production of the rabbitfish Siganus guttatus (Bloch) and the sea bass Lates calcarifer (Bloch), In: Juario, J.V., Benitez, L.V., (editors),
US
Proceedings of the Seminar on Aquaculture Development in Southeast Asia, 8-12 September 1987, Iloilo City, Philippines, SEAFDEC Aquaculture Department 195-210.
AN
Foroughifard, H., Tazikeh, E., Gharavi, B., Zarshenas, G.A., 2007. Artificial Reproduction Of Siganus Sutor Using Lhrha2 And HCG In Hormozgan Province, South Iran. Iranian Scientific Fisheries
M
Journal 16 (1), 161-168.
ED
Fourooghifard, H., Daghooghi, B., Aftabsavar, Y., 2009. Reproductive biology of the white spotted rabbit fish, Siganus sutor, in culture conditions (in persian). Iranian Scientific Fisheries Journal 18, 119-
PT
128.
CE
Fourooghifard, H., Matinfar, A., Abdolalian, E., Moezzi, M., Roohani, G.K., Kamali, E., Allen, S., Zahedi, M., 2017. Egg production and larval rearing of orange-spotted grouper (Epinephelus
AC
coioides) using reared broodstocks in Hormozgan Province, Iran. Iranian Journal of Fisheries Sciences 16 (3), 984-992. Hara, S., Duray, M.N., Parazo, M., Taki, Y., 1986. Year-round spawning and seed production of the rabbitfish, Siganus guttatus. Aquaculture 59, 259-272. Horký, P., Slavík, O., Bartos, L., Kolárová, J., Randák, T., 2006. The effect of the moon phase and seasonality on the behaviour of pikeperch in the Elbe River. Folia Zoologica 55, 411-417
ACCEPTED MANUSCRIPT Islam, M., Sultana, N., Hossain, M.B., Mondal, S., 2012. Estimation of fecundity and gonadosomatic index (GSI) of gangetic whiting, Sillaginopsis panijus (Hamilton, 1822) from the Meghna River Estuary, Bangladesh. World Applied Sciences Journal 17, 1253-1260. Juario, J.V., Duray, M.N., Duray, V.M., Nacario, J.F., Almendras, J.M., 1985. Breeding and larval rearing
T
of the rabbitfish, Siganus guttatus (Bloch). Aquaculture 44, 91-101.
IP
Kamukuru, A.T., 2006. Reproductive Biology of the White Spotted Rabbitfish, Siganus Sutor (Pisces:
CR
Siganidae) from Basket Trap Fishery in Dar Es Salaam Marine Reserves Systems, WIOMSA Tanzania, 60 p.
US
Khara, H., Oryan, S., 2012. Effects of LHRH-A2 and chlorpromazine (dopamine antagonists) on inducing spawning in Caspian Kutum, Rutilus frisii kutum, from the southwest of the Caspian Sea. Caspian
AN
Journal of Environmental Sciences 10, 33-42.
Kucharczyk, D., Kujawa, R., Mamcarz, A., Targońska-Dietrich, K., Wyszomirska, E., Glogowski, J.,
M
Babiak, I., Szabo, T., 2005. Induced spawning in bream (Abramis brama L.) using pellets containing
ED
GnRH. Czech Journal of Animal Science 50, 89-95. Lam, T., 1974. Siganids: their biology and mariculture potential. Aquaculture 3, 325-354.
PT
Lam, T., Soh, C., 1975. Effect of photoperiod on gonadal maturation in the rabbitfish, Siganus
CE
canaliculatus Park 1797. Aquaculture 5, 407-410. Marte, C., 1989. Hormone-induced spawning of cultured tropical finfishes. In Advances in Tropical
AC
Aquaculture, Workshop at Tahiti, French Polynesia, 20 Feb-4 Mar 1989, IFREMER, Actes de Colloque 9, 519- 539. Mylonas, C.C., Fostier, A., Zanuy, S., 2010. Broodstock management and hormonal manipulations of fish reproduction. General and Comparative Endocrinology 165, 516-534. Naeem, M., Zuberi, A., Salam, A., Ashraf, M., Elahi, N., Ali, M., Ishtiaq, A., Malik, T., Khan, M.J., Ayaz, M.M., 2011. Induced spawning, fecundity, fertilization rate and hatching rate of Grass carp (Ctenopharyngodon idella) by using a single intramuscular injection of ovaprimC at a fish hatchery in Faisalabad, Pakistan. African Journal of Biotechnology, 10, 11048-11053.
ACCEPTED MANUSCRIPT Nelson, S., G., Lock, S.A., Collins, L.A., 1992. Growth of the Rabbitfish Siganus Randalli Woodland in Relation to the Feasibility of Its Culture on Guam. University of Guam Marine Laboratory. Technical Report No. 97, Micronesia, 30p. Park Y.J., Takemura, A, Lee, Y.D., 2006. Lunar-synchronized reproductive activity in the pencil-streaked
T
rabbitfish Siganus doliatus in the Chuuk Lagoon, Micronesia. Ichthyological Research 53, 179-181.
IP
Popper, D., Gordin, H., Kissil, G.W., 1973. Fertilization and hatching of rabbitfish Siganus rivulatus.
CR
Aquaculture 2, 37-44.
Popper, D., May, R., Lichatowich, T., 1976. An experiment in rearing larval Siganus vermiculatus
US
(Valenciennes) and some observations on its spawning cycle. Aquaculture 7, 281-290. Popper, D., Pitt, R., Zohar, Y., 1979. Experiments on the propagation of Red Sea siganids and some
AN
notes on their reproduction in nature. Aquaculture 16, 177-181.
Rahman, M., Takemura, A., Park, Y., Takano, K., 2003. Lunar cycle in the reproductive activity in the
M
forktail rabbitfish. Fish Physiology and Biochemistry 28, 443-444.
ED
Rahman, M.S., Takemura, A., Takano, K., 2001. Lunar synchronization of testicular development and steroidogenesis in rabbitfish. Comparative Biochemistry and Physiology Part B: Biochemistry and
PT
Molecular Biology 129, 367-373.
CE
Randall, J.E., Allen, G.R., Steene, R.C., 1997. Fishes of the Great Barrier Reef and Coral Sea. University of Hawaii Press, Honolulu, Hawaii , 608 p.
AC
Soletchnik, P., 1984. Aspects of nutrition and reproduction in Siganus guttatus with emphasis on applications to aquaculture. Southeast Asian Fisheries Development Center, Iloilo, philippines. Takemura, A ,.Rahman, M., Nakamura, S., Park, Y.J., Takano, K., 2004. Lunar cycles and reproductive activity in reef fishes with particular attention to rabbitfishes. Fish and Fisheries 5, 317-328. Takemura, A., Rahman, M., Park, Y., 2010. External and internal controls of lunar-related reproductive rhythms in fishes. Journal of Fish Biology 76, 7-26.
ACCEPTED MANUSCRIPT Zohar, Y., Mylonas, C.C., 2001. Endocrine manipulations of spawning in cultured fish: from hormones to genes, In: Lee , C. S. (Edtor), Reproductive Biotechnology in Finfish Aquaculture. Elsvier, Netherlands, pp. 99-136.
T
Fig. 1 Egg production of Siganus. sutor on various days of reproduction season (the numbers in
IP
parentheses show the lunar days) (Mean ±Se).
CR
Fig. 2 Egg production of S. sutor treated by different hormone administrations (Mean ±Se).
US
Fig. 3 The regression between relative fecundity and total body weight of S. sutor treated by
AN
hormone administration.
Fig. 4 Dendrogram of hierarchical analysis of average egg production in S. sutor treated by
M
different hormone administrations during the reproduction season.
ED
Fig. 5. Dendrogram of hierarchical analysis of average egg production in S. sutor treated by
PT
different hormone administrations.
Date of hormone injection May 8 (29)
May 15 (7)
May 22 (14)
June 1 (22)
June 8 (1)
June 15 (8)
Mean ± SE
0
0
0
66.67
66.67
0
0
0
16.7±10.9
0
100
100
100
100
100
66.7
0
70.8±16
H
0
33.3
66.7
100
100
33.3
33.3
0
45.8±14
HM
0
33.3
66.7
100
100
33.3
33.3
0
45.8±14
C
0
0
0
33.3
0
0
0
0
4.2±4.2
Mean ± SE
0
33.3±18.3
46.7±2
80±13.3
73.3±19.4
33.3±18.3
26.7±12.5
0
L1 L2
AC
May 1 (22)
Treatment
April 12 (3)
CE
Table 1 Total spawned females (%) of S. sutor treated by all types of hormone administration on various days of reproductive season (the numbers in parentheses show the lunar days) (Mean ± Se).
ACCEPTED MANUSCRIPT L1= LHRHa2 hormone at dosage of 10 μg/Kg body weight (BW) ; L2 = LHRHa2 hormone at dosage of 20 μg/Kg BW ; H= HCG hormone at a dosage of 2000 IU/kg BW ; HM = HCG hormone at a dosage of 2000 IU/kg BW combined with 5 mg metoclopramide/Kg BW ; C = 1 ml 0.9% saline water.
Highlights: The information of the present study can be helpful to fish breeders in planning for the most efficient egg production and rearing of this important species.
T
IP
A significantly greater number of spawned fish (70.83 ± 9.48 %), was observed in treatment L2
CR
US AN M ED PT CE AC
compared with the other treatments (L1: 16.67±7.77 %, H: 41.67± 10.28 %, HM: 41.67±10.28 % and C: 4.17± 4.17 %). Cluster analysis showed that the best time for induced spawning of S. sutor was 15 to 22 May and the best hormone for induced spawning was LHRHa2 at a dosage of 20µg/kg BW of females.