Reproductive Performance of the First Generation Stichopus horrens Broodstock

Reproductive Performance of the First Generation Stichopus horrens Broodstock | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Reproductive Performance of the First Generation Stichopus horrens Broodstock Sari Budi Moria Sembiring, Ketut Maha Setiawati, Jhon Harianto Hutapea, and 6 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6712637/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 18 Dec, 2025 Read the published version in Scientific Reports → Version 1 posted 8 You are reading this latest preprint version Abstract The production of cultivated sea cucumber Stichopus horrens broodstock is urgently being carried out, considering the sharp decline of natural broodstock. Therefore, natural broodstock (F0) were spawned, and their larvae were reared in a hatchery until they metamorphosed into seeds (F1). Further cultured in the earthen pond for 16 months to reach broodstock size. After determining the sex by the eviceration method, males and females were cultured in separate concrete tanks measuring 2x1x1m 3 to regenerate internal organs and gonad maturation, and fed with fresh grounded Sargassum sp. and Ulva sp. at 3% biomass. Eleven months later, the F1 broodstock was divided into three groups based on body weight: small (116.22 ± 9.81 g and 109.80 ± 5.72 g), medium (134.08 ± 7.48 g and 136.10 ± 12.78 g), and large (152.56 ± 190.01 g and 170.20 ± 7.92 g) for male and female, respectively. Five pairs from each broodstock group were spawned separately in fifteen 30 L polycarbonate tanks. The F1 broodstock could produce 180,000 to 820,000 eggs per broodstock, but not significantly different ( P > 0.05) among broodstock size groups. The highest egg diameter (200.92 ± 18.18 µm) and larval survival were obtained in the large broodstock size group and were significantly different ( P < 0.05) from the others. The highest hatching rate (82.12 ± 4.38%) was obtained in large broodstock size, followed by medium broodstock size, and was significantly different ( P < 0.05) from the small broodstock size group. Mean F1 is suitable for domesticated broodstock, especially from large broodstock size groups. Biological sciences/Biological techniques Biological sciences/Developmental biology Biological sciences/Physiology broodstock size cultivation performance reproduction Stichopus horrens Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction Sea cucumbers are marine invertebrates with high commercial value. One species that fishermen widely catch is Stichopus sp., locally named Gamat. This sea cucumber has a high collagen content and various metabolite compounds such as amino acids, polysaccharides, triterpene glycosides, carotenoids, vitamins, minerals, phenolic compounds, bioactive compounds, peptides, chondroitin sulfate, and saponins (Sottoroff et al. 2013 ; Pangestuti and Arifin 2018 ; Rahael et al. 2019 ; Xing et al. 2021 ; Yang et al. 2018 ). This has resulted in a decline in sea cucumber populations in nature. The imbalance between the level of sea cucumber captured and the low recruitment rate will accelerate the extinction of the sea cucumber population in its habitat. Therefore, to anticipate the extinction of sea cucumbers in nature, in this case Stichopus sp., it is necessary to produce seeds from the hatchery. This seedling has been carried out in the Gondol Marine Biota Scientific Conservation Area (MBSCA, Gondol) since 2022. It has shown significant progress, with larval survival to stadia doliolaria by 12–22%, which then metamorphoses into seeds (Sembiring et al. 2025 ). Lately, the availability of natural broodstock has become increasingly limited, so the sustainability of the hatchery is expected to be carried out using domesticated broodstock. In aquaculture, the core of domestication is the total control of an organism's life cycle, including its breeding activities (Liao and Huang 2000 ). Perfect domestication is when the entire life cycle of aquatic biota can be maintained in the cultivation system. The commodity in this study is Stichopus sp. The domestication carried out is almost perfect, where wild broodstock has been able to adapt to a new environment in captivity and controlled (Widiastuti et al. 2019 ), can utilize the feed provided so that it can grow and even be able to spawn and produce seeds (Setiawati et al., 2021 ). In shrimp, the weight and age of the broodstock are targeted in the domestication process because they affect reproductive performance (Coman and Crocos 2003 ; Nur et al. 2022 ; Racotta et al. 2003 ). Generally, large broodstock tend to have better reproductive performance than small ones (Hoang et al. 2002 ). However, research has not been reported on the reproductive performance of Stichopus sp. cultivated F1 broodstock and the development of the larvae they produce. Therefore, this study was conducted to obtain information on the reproductive performance of Stichopus sp. F1 broodstock in different size groups are used for hatchery and seed production activities. Materials and Methods Maintenance of cultivated F1 broodstock The Stichopus horrens F1 broodstock used in this study came from seeds produced in the hatchery of MBSCA, Gondol. The seeds cultivated in the pond are selected solely based on size, specifically 13.31 ± 3.17 g. They were then raised in an experimental earthen pond of the Institute for Mariculture Research and Fisheries Extension (IMRAFE), Gondol, Bali. Although cultivated under the same environmental conditions and fed for 16 months, they exhibit varied growth (54–151 g). At this size, from the same genus, S. chloronotus , the gonads have formed (Hoareau and Conand 2001 ), so this size is categorized as potential broodstock and used for sex determination. Sex determination was conducted for 121 potential broodstock by injecting 0.2 µL of 0.5 M Potassium chloride solution (CAS 7447-40-7 Merck). Sixty-five were found as male with a length of 14.4 ± 2.7 cm and a weight of 77.8 ± 22.4 g, and 56 were female with a length of 14.5 ± 2.7 cm and a weight of 88.7 ± 28.2 g. Seventeen unknown sex with a length of 13.38 ± 2.3 cm and a weight of 45.7 ± 20.9 g were not used for further cultivation. After the sex was determined, the male and female were kept separately in two 2x1x1 m 3 concrete tanks, fed with fresh grounded Sargassum sp. and Ulva sp., in a 2:1 ratio, 3% of the total sea cucumber biomass for the regeneration of internal organs and gonad maturation. In addition to the two feeds, the sea cucumbers eat benthos that grow on the walls and bottom of the tank. After 11 months of culture, they have reached a length of 17.9 ± 5.0 cm and a weight of 156.7 ± 52.2 g for males and 17.7 ± 5.2 cm and 137.7 ± 27.5 g for females, and are ready to be used as test animals in this study. Spawning of cultivated broodstock Before spawning, the F1 broodstock was measured, weighed, and divided into three size groups, namely small (S), medium (M), and large (L). The average length and weight of the male broodstock from the small size group (13.18 ± 2.03 cm; 116.22 ± 9.81g), medium (15.0 ± 2.15 cm; 134.08 ± 7.48 g), and large (16.46 ± 3.28 cm; 152.56 ± 19.01 g) groups. The average length and weight of the female broodstock for the small size group (12.40 ± 1.14 cm; 109.80 ± 5.72 g), medium (14.20 ± 1.25 cm; 136.10 ± 12.78 g), and large (15.90 ± 2.56 cm; 170.20 ± 7.92 g) groups. After grouping small, medium, and large broodstock categories, five pairs were found to represent each group. According to their group, each female was placed in a polycarbonate container with a volume of 30 L. Then, the male was placed in a plastic basket and put into the container to prevent the male and female from mixing (Fig. 1 ). Each size group had five replicates, so the number of spawners was 15 pairs. Spawning is carried out for two consecutive days during the new moon period. The seawater used for spawning passes through a sand filter and a 5 µm cartridge filter (Purerite PS-05, KEMFLO). After filtration, the seawater is stored in a concrete tank, sterilized using two 35-watt UV lamps (UV Lamp SAKKAI PRO 35 WATT) for 16 hours, and is ready to transfer into the spawning tank. Each spawning tank is covered with black plastic to maintain a stable temperature. The spawning is carried out for 2 days. Larvae rearing Eggs produced from each pair of broodstock size groups are harvested and incubated for 24 hours in 15 units of 100 L black plastic tanks according to treatment. After hatching, the larvae are stocked at a density of 200 individuals L − 1 in 15 units of an 80 L larval rearing tank. Larvae were fed with Chaetoceros muelleri, Isochrysis galbana, Nitzchia sp., and Navicula sp. according to their development (Table 1 ). Table 1 Feeding scheme for Stichopus horrens larval rearing until they reach the juvenile stage Day 1–6 7–12 13–21 22–45 Chaetoceros muelleri 10–20 x 10 4 cell ml -1 Isochrysis galbana 0.5–10 x 10 4 cell ml -1 Nitzchia sp. 0.5–15 x 10 4 cell ml -1 Navicula sp. 0.1–1.75 x 10 4 cell ml -1 Water replacement starts on the fifth day by slowly lowering the water by 20% and then adding UV-sterilized water using a plankton net. The water is changed every two days. Water quality monitoring, including temperature, is carried out daily, while pH, dissolved oxygen, ammonia, and nitrite are monitored twice a week. After 40 days of rearing, larvae metamorphosed into juveniles, ready to harvest, and were kept in a nursery tank. Data Analysis The variables observed include the length and weight of the broodstock, the number of broodstock spawning, the number of eggs, the egg diameter, the hatching rate, the development of the larvae, the survival rate, and the proportion of large, medium, and small juveniles at the end of the study. Egg diameter and total length of larvae were measured using a microscope connected to a camera (Nikon DXM 1200F) and Win-Roof ver. 5.0 software. The normality and homogeneity of all data sets were checked through the Non-Parametric One-Sample Kolmogorov-Smirnov and Levene Test, respectively (SPSS v. 20, IBM, Armonk, NY, USA). Differences among treatments were detected in One-Way ANOVA, followed by post hoc Tukey HSD. Data are expressed as mean SE of the mean (n = 10–15). The significance level was 0.05. Results Spawning of F1 broodstocks, fecundity, egg diameter, and hatching rate In two days of spawning trials, three pairs of broodstock spawned in each small, medium, and large size group with an average number of eggs: 489,600 ± 202,413, 420,000 ± 217,607, and 684,800 ± 215,556 eggs, respectively. Statistically, the number of eggs from the three broodstock size groups was not significantly different ( P > 0.05) (Table 2 ). The egg diameters in all size groups differed significantly (F (2,6) = 22.149, P = 0.002), with the highest average found in the large size group (200.92 ± 18.18 µm), followed by the medium and small size groups of 174.22 ± 15.61 and 137.65 ± 12.47 µm, respectively. Meanwhile, the average hatching rate in the large size group (82.12 ± 5.37%) was not significantly different ( P > 0.05) from the medium size group (67.84 ± 7.64) but significantly different from the small size group (26.29 ± 8.52) (F (2,6) = 47.350, P = 0.000). Table 2 Reproductive performance (number of eggs, egg diameter, and hatching rate) of F1 broodstock Stichopus horrens in different size groups Size Groups Body weight of broodstock (g) Number of eggs (eggs/ind) Egg diameter (µm) Hatching rate (%) Male Female Small 124.8 117.0 722,400 132.61 ± 8.32 34.56 105.0 105.3 355,200 125.59 ± 11.87 17.54 125.4 115.0 391,200 154.76 ± 17.23 26.78 Average 118.40 ± 9.50 a 112.40 ± 5.10 a 489,600 ± 202,413 a 137.65 ± 12.47 a 26.29 ± 6.96 a Medium 145.0 129.0 609,600 163.77 ± 11.58 79.40 130.8 125.5 182,400 182.17 ± 18.15 69.82 127.1 158.0 468,000 176.73 ± 17.09 84.30 Average 134.30 ± 7.70 b 137.50 ± 14.57 b 420,000 ± 217,607 a 174.22 ± 15.61 b 77.84 ± 6.01 b Large 185.6 183.0 794,400 184.05 ± 16.19 85.26 150.6 166.0 436,800 201.40 ± 19.85 75.92 141.5 162.0 823,200 217.31 ± 18.51 85.18 Average 159.20 ± 19.00 c 170.33 ± 9.10 c 684,800 ± 215,556 a 200.92 ± 18.18 c 82.12 ± 4.38 b Different alphabetic letters between the averages in the same column indicate statistically significant ( P < 0.05). Larval development, survival rate, and percentage of juveniles Larval development was not different in broodstock of different sizes. Early auricularia is characterized by a well-visible functional mouth, esophagus, cloaca, and ciliated band used for feeding and movement. Final auricularia is reached 13–15 days after hatching, and on the 17th − 21st day, the larvae reach the doliolaria stage. Approaching the metamorphosis to the pentactula, the behaviour changes from planktonic to benthic larvae, the ciliated band disappears, five tentacles are visible, and the larvae move to the bottom of the rearing tank, occurring after 21 days of rearing (Figs. 2 and 3 ). The survival of larvae 40 days post-fertilization was significantly different (F (2,6) = 35.816, P = 0.000) between large-size groups and medium and small broodstock groups. Meanwhile, the medium and small-size broodstock groups did not differ significantly ( P > 0.05). The survival rate pattern of larvae for each size broodstock group is shown in Fig. 4 . The water quality parameters in the larval rearing medium, including temperature, pH, dissolved oxygen, ammonia, and nitrite, are presented in Table 3 and are still feasible to support sea cucumber larvae rearing. Table 3 Average water quality measurement results during larval rearing Parameters Small-size group Medium-size group Large-size group Temperature (°C) 29.08 ± 0.09 29.03 ± 0.07 29.13 ± 0.11 pH 8.26 ± 0.03 8.26 ± 0.01 8.27 ± 0.01 Dissolved oxygen (ppm) 5.30 ± 0.07 5.25 ± 0.07 5.27 ± 0.08 Ammonia (ppm) 0.12 ± 0.02 0.10 ± 0.03 0.08 ± 0.02 Nitrite (ppm) 0.09 ± 0.01 0.08 ± 0.03 0.06 ± 0.01 Figures 5 and 6 show the average length and weight of juvenile S. horrens and the percentage of juvenile size groups. The analysis results show that each size group's body length, weight, and percentage are not significantly different ( P > 0.05). Discussion Spawning of F1 broodstock, fecundity, egg diameter, and hatching rate Two-year-old S. horrens broods have succeeded in spawning for the three size groups studied, with a range of sizes; even small female and male broodstock groups with an average body weight of 105–125 g can spawn, as well as broodstock in large and medium groups. The broodstock used in this research is much larger than other species. The first gonad maturity in Actinopyga echinites was detected at a total weight of 65 g (Kohler et al. 2009 ), and Sembiring et al. ( 2018 ) reported that F1 broodstock H. scabra can spawn at an average body weight of 122.6 ± 32.37 g. This indicates that the nutritional reserves of the broodstock play an important role and can improve reproductive performance. The quantity and quality of these nutrient reserves determine the fecundity and quality of the eggs, as also mentioned by Schreck et al. ( 2001 ). Thus, the energy source required for embryo and larval development depends mainly on the broodstock's nutrient reserves (Huang et al. 2010 ). The three-size broodstock groups show the same spawning frequency, and the number of eggs produced did not differ significantly. These results show that the weight of the brooder does not always affect the number of eggs produced, especially because there is a large difference in the number of eggs between individuals in one brood size group. Given that the broodstock used in this experiment is the same age, the maintenance from seed to broodstock is done under the same environmental and feed conditions. Thus, it can be concluded that if broodstock rearing is optimal, then size is not the main factor in the success of spawning. This is in line with Gianasi ( 2018 ), who reports that in Cucumaria frondosa , optimal environmental conditions can increase egg reproduction in addition to the broodstock's size. Furthermore, apart from environmental conditions, other external factors that can affect broodstock productivity include the type of feed given, the maintenance container, the phase of the moon, and the season (photoperiod and temperature) ( Hu et al. 2013 ; Laguerre et al. 2020 ; Rakaj et al. 2017 ; Tehranifard et al. 2007). Ru et al. ( 2021 ) also reported that sea cucumbers use nutritional sources to support gonad growth, so their reproductive performance depends on the availability and quality of food. The feed given to the broodstock during maintenance is ground Sargassum sp. and Ulva sp. In addition, the broodstock also eats benthos that grow naturally in the broodstock maintenance tank. Moreover, Wen et al. ( 2016 ) reported that A. japonicus and S. horrens prefer Sargassum sp. as food. Xu et al. ( 2022 ) also reported that the food source for Stichopus monotuberculatus comes from Sargassum sp., phytoplankton, and organic matter. The average diameter of eggs from the three size groups of F1 broodstock S. horrens was significantly different ( P < 0.05 ), with the highest average egg diameter (217.31 ± 18.51 µm) obtained from the large size group broodstock. Conand ( 1993 ) reported that the Stichopus variegatus begins maturing at a length of 270 mm and with an oocyte diameter of 180 µm, but its fecundity is only 10,000 oocytes. The egg diameter in this experiment is far bigger, even compared to the small-size group (132–154 µm). De La Rosa et al. ( 2023 ) s tate that the egg's large diameter indicates a more significant food reserve that can support the development of the embryo and larvae when hatching, making these conditions excellent for the survival of sea cucumber larvae. The egg-hatching rates of the large and medium groups differed significantly ( p < 0.05) from the small groups. The highest hatching rate is obtained from the large group, followed by the medium-sized group, so the lowest hatching rate is in the small group. The condition of the female broodstock can affect egg quality and hatching rate. Marquet et al. ( 2017 ) reported that the egg-hatching rates of Holothuria arguinensis and H. Mamata were influenced by egg quality, fertility rates, and environmental conditions. The same has been reported in sea urchins Paracentrotus lividus (Guettaf et al. 2000 ). During spawning and egg incubation, salinity and temperature were maintained at 33–34 ppt and 30–31°C for all broodstock size groups, and these ranges are optimum for most sea cucumbers, as reported by Hu et al. ( 2013 ). The salinity level is one of the significant environmental factors affecting the hatching rate. Tu et al. ( 2022 ) reported that the Japanese sea cucumber Apostichopus japonicus can produce a high hatching rate at a salinity of 34 ppt. Asha et al. ( 2011 ) also noted that the optimal salinity range for hatching H. scabra eggs and rearing the larvae to the auricularia stage is 33–35 ppt. Furthermore, Falconer and Mackay ( 1996 ) also state that the phenotype of an organism is influenced by its genotype, environment, and interaction. In this study, large broodstock groups resulted in higher hatching rates. This suggests that the size of sea cucumber broodstock affects the hatching rate of eggs, and it can be assumed that this hatching rate is primarily an inherited trait. Compared to the performance of the natural broodstock, which has a high egg production of 1–2 million eggs, the egg production of the F1 broodstock is still much lower, at 400,000-600,000 eggs. Environmental conditions in nature, which vary the availability of food, result in a higher fecundity of natural broodstock than domesticated broodstock, which is kept in tanks and fed a limited type of food. This difference in fecundity is likely related to higher nutrition due to the availability of more varied foods during broodstock rearing in ponds. During rearing in the concrete tank for the regeneration of the gastrointestinal tract and gonads, gonad maturation, and spawning of the broodstock, the feed given is limited to Sargassum sp., Ulva sp., and benthos that grow on the inner wall of the tank. Therefore, research on natural and artificial feeds for domesticated broodstock still needs to be continued. Cong et al. ( 2024 ) state that the variation in fecundity in sea cucumbers is related to the difference in the amount of food consumed by sea cucumbers. Larval development, survival rate, and percentage of juveniles The development of sea cucumber larvae from different broodstock size groups did not differ significantly ( p > 0.05). Larvae of S. horrens of large broodstock size have a high survival rate and significantly differ from medium and small broodstock sizes. In addition, larger sizes have more energy sources to provide nutrients for the development of embryos and larvae when hatching, making this condition very favourable for the survival of larvae (Widyastuti et al. 2008 ). This can be seen from the diameter of the eggs produced by large-sized broodstock, which is more significant than the diameter of the eggs in other broodstock size groups. Morgan ( 2009 ) reports that Australostichopus mollis larvae from larger-sized broodstock tend to have better access to food sources in the early developmental phase, which supports increased growth and larval survival. Peters-Didier and Sewell ( 2008 ) reported a relationship between egg size and lipid content, as the lipid content in eggs is a significant energy reserve in the early development of Australostichopus mollis larvae. This makes it possible for larger broodstock sizes to provide more energy to each egg, which in turn has the potential to produce better larvae. Some research results state that the larval development phase of the S. horrens sea cucumber is the same as that of other Aspidochirotes sea cucumbers (Chen and Chian 1990 ; Hu et al. 2010 ; McEuen 1988 ; Ramofafia et al. 2003 ). The food and environmental factors during larval rearing significantly affect juvenile survival (Yu et al. 2022 ). Water quality is important in a sea cucumber hatchery because larvae are susceptible to environmental changes (Asha and Muthiah 2006 ). This study's water temperature, pH, dissolved oxygen, ammonia, and nitrite were relatively consistent and generally within the normal range for maintaining sea cucumber larvae of tropical holothurian species (Hu et al. 2010 ; Knauer 2011 ). Conclusion The first generation of domesticated S. horrens broodstock from the large broodstock group (170.33 ± 9.10 g) provides better reproduction and larval performance. Declarations Author contribution Sari Budi Moria Sembiring and Ketut Maha Setiawati joined in all activities, from arranging the idea and designing the experiment, broodstock, larval and juvenile rearing, samples collection, larval measurement, statistical analysis, data interpretation, and preparation of the original draft. Jhon Harianto Hutapea, Gunawan Gunawan, Ananto Setiadi, and Ni Wayan Widya Astuti joined in conducting broodstock, larval and juvenile rearing, sample collection, larval measurement, and statistical analysis. Haryanti Haryanti and Nyoman Adiasmara Giri arranged the idea, designed the experiment, performed data interpretations, and prepared the original draft. Rarastoeti Pratiwi performed data interpretations and prepared the original draft. All authors have read and agreed to the published version of the manuscript . The Ethics Commission for Animal Care and Use of the Indonesian National Research and Innovation Agency decided that this study does not require Ethics Clearance, and the animals in this study are invertebrates that are exempt from Ethics Clearance (Reference Number: 215/KE.02/SK/12/2023). Funding This research was funded by the Directorate of Research and Innovation Funding, Deputy for Facilitation of Research and Innovation, National Research and Innovation Agency (No. 61/II.7/HK/2024). Competing interest The authors declare no conflicts of interest regarding this research article. Acknowledgments The authors are grateful to all technicians at the sea cucumber hatchery for their technical assistance during the experiment and to the Directorate of Research and Innovation Funding, Deputy for Facilitation of Research and Innovation, National Research and Innovation Agency. References Asha, P. S. & Muthiah, P. 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Revista de Biología Mar. y Oceanografía . 48 (1), 23–35. https://doi.org/10.4067/S0718-19572013000100003 (2013). Tehranifard, A. et al. Reproductive cycle of Stichopus herrmanni from Kish Island, Iran. SPC Beche-de-mer Information Bulletin. 24 : 22–27. (2006). Tu, P. T. C. et al. Impact of short-term salinity and turbidity changes on hatching and survival rates of Japanese sea cucumber, Apostichopus japonicus (Selenka, 1867), Eggs. Asian Fisheries Science. 35: 90–94. (2022). https://doi.org/10.33997/j.afs.2022.35.1.008 Wen, B. et al. Effects of dietary inclusion of benthic matter on feed utilization, digestive and immune enzyme activities of sea cucumber Apostichopus japonicus (Selenka). Aquaculture 458 , 1–7. https://doi.org/10.1016/j.aquaculture.2016.01.028 (2016). Widiastuti, Z. et al. Utilization of seaweed to support sea cucumber domestication activities ( Stichopus sp.). In Proceedings of the XVI Annual National Seminar on Fisheries and Marine Research, Department of Fisheries, Faculty of Agriculture, Gadjah Mada University, Yogyakarta. pp. 50–54. (2019). Widyastuti, Y., Subagja, J. & Gustiano, R. Reproduction of selected and non-selected nile tilapia ( Oreochromis niloticus ) with artificial induced breeding: character of broodstock, egg, embryo, and larvae. Jurnal Iktiologi Indonesia . 8 (1), 1–20. https://doi.org/10.32491/jii.v8i1.282 (2008). Xing, R. et al. Authentication of sea cucumber products using NGS-based DNA mini-barcoding. Food Control . 129 , 108199. https://doi.org/10.1016/j.foodcont.2021.108199 (2021). Xu, Q. et al. Sea ranching feasibility of the hatchery-reared tropical sea cucumber Stichopus monotuberculatus in an inshore coral reef island area in South China Sea (Sanya, China). Front. Mar. Sci. 9 , 918158. https://doi.org/10.3389/fmars.2022.918158 (2022). Yang, L. et al. Separation, purification, structures, and anticoagulant activities of fucosylated chondroitin sulfates from Holothuria scabra . Int. J. Biol. Macromol. 108 , 710–718. https://doi/org/10.1016/j.ijbiomac.2017.11.058 (2018). Yu, Z. et al. Effects of diet on larval survival, growth, and development of the sea cucumber Holothuria leucospilota . Aquaculture Nutr. Article ID . 8947997. https://doi.org/10.1155/2022/8947997 (2022). Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 18 Dec, 2025 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 12 Aug, 2025 Reviews received at journal 06 Jul, 2025 Reviewers agreed at journal 09 Jun, 2025 Reviewers invited by journal 27 May, 2025 Editor assigned by journal 27 May, 2025 Editor invited by journal 27 May, 2025 Submission checks completed at journal 23 May, 2025 First submitted to journal 23 May, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6712637","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":462826703,"identity":"0c43247d-de45-4386-9f4f-624fa60e1774","order_by":0,"name":"Sari Budi Moria Sembiring","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzElEQVRIie3QIQ+CQBTA8XdjO8qb+TY3+ArnDBY+zF05Et1ogsKg6regORruKnYiFjLROaaixZkOmpv3b2+73/beAdhsP1oltg+PA9m9JwpiAulrZz2PkEPsyOIzG4ifnC8aKQ2PSx23cAtg4RoIr0OuETEqM5lwkimg2BoIKNDIWFTUJGYk1UCZabG8GwnnIZ9MoFFw2gsh3gSuEwhvuvGTq2pVpiORO4XGW/xcOb28V/4G3Y71Q+D5iWmxr2QMOOf9q2EusNlstn/oCfjWQdzUzAGhAAAAAElFTkSuQmCC","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":true,"prefix":"","firstName":"Sari","middleName":"Budi Moria","lastName":"Sembiring","suffix":""},{"id":462826704,"identity":"70019165-16a4-4ce3-ad4a-df0da1f2b93c","order_by":1,"name":"Ketut Maha Setiawati","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Ketut","middleName":"Maha","lastName":"Setiawati","suffix":""},{"id":462826705,"identity":"d2af0d6b-c0c4-4f99-b4da-0e2b7789e291","order_by":2,"name":"Jhon Harianto Hutapea","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Jhon","middleName":"Harianto","lastName":"Hutapea","suffix":""},{"id":462826706,"identity":"fed211dc-e5b4-40bf-adfa-e0465bac3dd8","order_by":3,"name":"Gunawan Gunawan","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Gunawan","middleName":"","lastName":"Gunawan","suffix":""},{"id":462826707,"identity":"eafb1c80-a348-4819-bfd6-f52291d6dc7e","order_by":4,"name":"Ananto Setiadi","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Ananto","middleName":"","lastName":"Setiadi","suffix":""},{"id":462826708,"identity":"6167e3fb-78c1-46cd-bb0f-d48c372bdc9a","order_by":5,"name":"Ni Wayan Widia Astuti","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Ni","middleName":"Wayan Widia","lastName":"Astuti","suffix":""},{"id":462826709,"identity":"a9a9f9a3-83fa-4007-aa4d-ee08b8f0770e","order_by":6,"name":"Haryanti Haryanti","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Haryanti","middleName":"","lastName":"Haryanti","suffix":""},{"id":462826710,"identity":"0de41150-35ea-48ee-bbf4-3f98afa52bf6","order_by":7,"name":"Nyoman Adiasmara Giri","email":"","orcid":"","institution":"National Research and Innovation Agency","correspondingAuthor":false,"prefix":"","firstName":"Nyoman","middleName":"Adiasmara","lastName":"Giri","suffix":""},{"id":462826711,"identity":"00f3a680-6cc8-4946-a683-8a52ad3ecf75","order_by":8,"name":"Rarastoeti Pratiwi","email":"","orcid":"","institution":"Gadjah Mada University","correspondingAuthor":false,"prefix":"","firstName":"Rarastoeti","middleName":"","lastName":"Pratiwi","suffix":""}],"badges":[],"createdAt":"2025-05-21 05:23:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6712637/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6712637/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1038/s41598-025-30858-w","type":"published","date":"2025-12-18T15:56:56+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":83686668,"identity":"3aedab25-347a-47c6-a788-d82db09e7e0e","added_by":"auto","created_at":"2025-05-30 18:31:40","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":72617,"visible":true,"origin":"","legend":"\u003cp\u003eSpawning container (male broodstock in a plastic basket and female broodstock directly in a polycarbonate tank).\u003c/p\u003e","description":"","filename":"Picture1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/5c4ecdf718ea8a96ac8d3f66.jpg"},{"id":83686671,"identity":"2cb7e341-8d8a-42d0-947a-03cd2b61d3c1","added_by":"auto","created_at":"2025-05-30 18:31:40","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":134526,"visible":true,"origin":"","legend":"\u003cp\u003eLarval development of \u003cem\u003eStichopus horrens\u003c/em\u003e (A1 Early; A2 Middle; A3 Late Auricularia stage; B Doliolaria stage; and C Pentactula stage).\u003c/p\u003e","description":"","filename":"Picture2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/8626e4e597562378126f8c70.jpg"},{"id":83687041,"identity":"7d8e6628-4ffe-429d-8eee-7e4757e1b39c","added_by":"auto","created_at":"2025-05-30 18:39:40","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":126029,"visible":true,"origin":"","legend":"\u003cp\u003eLength growth of \u003cem\u003eStichopus horrens\u003c/em\u003e larvae from F1 broodstock of different size groups (different dot colors related to larvae from different broodstock size groups; dot line, dash line, and dot-dash trend line belong to larvae from large, medium, and small broodstock size groups consecutively).\u003c/p\u003e","description":"","filename":"Picture3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/42c580dbacff4291f9124441.jpg"},{"id":83687042,"identity":"4eed0f6b-0cfd-42eb-981c-cfbe509bf44c","added_by":"auto","created_at":"2025-05-30 18:39:40","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":119492,"visible":true,"origin":"","legend":"\u003cp\u003eThe survival rate of \u003cem\u003eStichopus horrens\u003c/em\u003e larvae from F1 broodstock of different size groups (different dot colors related to larvae from different broodstock size groups; dot line, dash line, and dot-dash trend line belong to larvae from large, medium, and small broodstock size groups consecutively).\u003c/p\u003e","description":"","filename":"Picture4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/416a9fb7359845e186c2ac04.jpg"},{"id":83687045,"identity":"d7fbf033-fe72-4a4a-8156-05ed8fd2c0c5","added_by":"auto","created_at":"2025-05-30 18:39:40","extension":"jpg","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":169204,"visible":true,"origin":"","legend":"\u003cp\u003eThe average length and weight of \u003cem\u003eStichopus horrens\u003c/em\u003e juveniles produced from F1 broodstock of different size groups.The proportion of \u003cem\u003eStichopus horrens\u003c/em\u003e juvenile size produced from Large, Medium, and Small F1 broodstock\u003c/p\u003e","description":"","filename":"Picture5.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/5035c7a52734a120c99b6cad.jpg"},{"id":83686675,"identity":"45e42ee6-4653-4e2e-8f5a-c5028186938c","added_by":"auto","created_at":"2025-05-30 18:31:40","extension":"jpg","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":274858,"visible":true,"origin":"","legend":"\u003cp\u003eThe proportion of \u003cem\u003eStichopus horrens\u003c/em\u003e juvenile size produced from Large, Medium, and Small F1 broodstock.\u003c/p\u003e","description":"","filename":"Picture6.jpg","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/9a4c862f1f447c63b77875e8.jpg"},{"id":98813822,"identity":"6d09f4be-6f9a-4c5f-a66e-cfc407070557","added_by":"auto","created_at":"2025-12-22 16:01:30","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1793380,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6712637/v1/9f13a71d-0bb5-406d-9eca-303bbb7c72dc.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Reproductive Performance of the First Generation Stichopus horrens Broodstock","fulltext":[{"header":"Introduction","content":"\u003cp\u003eSea cucumbers are marine invertebrates with high commercial value. One species that fishermen widely catch is \u003cem\u003eStichopus\u003c/em\u003e sp., locally named Gamat. This sea cucumber has a high collagen content and various metabolite compounds such as amino acids, polysaccharides, triterpene glycosides, carotenoids, vitamins, minerals, phenolic compounds, bioactive compounds, peptides, chondroitin sulfate, and saponins (Sottoroff et al. \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Pangestuti and Arifin \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Rahael et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Xing et al. \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Yang et al. \u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThis has resulted in a decline in sea cucumber populations in nature. The imbalance between the level of sea cucumber captured and the low recruitment rate will accelerate the extinction of the sea cucumber population in its habitat. Therefore, to anticipate the extinction of sea cucumbers in nature, in this case \u003cem\u003eStichopus\u003c/em\u003e sp., it is necessary to produce seeds from the hatchery.\u003c/p\u003e \u003cp\u003eThis seedling has been carried out in the Gondol Marine Biota Scientific Conservation Area (MBSCA, Gondol) since 2022. It has shown significant progress, with larval survival to stadia doliolaria by 12\u0026ndash;22%, which then metamorphoses into seeds (Sembiring et al. \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2025\u003c/span\u003e). Lately, the availability of natural broodstock has become increasingly limited, so the sustainability of the hatchery is expected to be carried out using domesticated broodstock. In aquaculture, the core of domestication is the total control of an organism's life cycle, including its breeding activities (Liao and Huang \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePerfect domestication is when the entire life cycle of aquatic biota can be maintained in the cultivation system. The commodity in this study is \u003cem\u003eStichopus\u003c/em\u003e sp. The domestication carried out is almost perfect, where wild broodstock has been able to adapt to a new environment in captivity and controlled (Widiastuti et al. \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), can utilize the feed provided so that it can grow and even be able to spawn and produce seeds (Setiawati et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In shrimp, the weight and age of the broodstock are targeted in the domestication process because they affect reproductive performance (Coman and Crocos \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Nur et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Racotta et al. \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). Generally, large broodstock tend to have better reproductive performance than small ones (Hoang et al. \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). However, research has not been reported on the reproductive performance of \u003cem\u003eStichopus\u003c/em\u003e sp. cultivated F1 broodstock and the development of the larvae they produce. Therefore, this study was conducted to obtain information on the reproductive performance of \u003cem\u003eStichopus\u003c/em\u003e sp. F1 broodstock in different size groups are used for hatchery and seed production activities.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMaintenance of cultivated F1 broodstock\u003c/h2\u003e \u003cp\u003eThe \u003cem\u003eStichopus horrens\u003c/em\u003e F1 broodstock used in this study came from seeds produced in the hatchery of MBSCA, Gondol. The seeds cultivated in the pond are selected solely based on size, specifically 13.31\u0026thinsp;\u0026plusmn;\u0026thinsp;3.17 g. They were then raised in an experimental earthen pond of the Institute for Mariculture Research and Fisheries Extension (IMRAFE), Gondol, Bali. Although cultivated under the same environmental conditions and fed for 16 months, they exhibit varied growth (54\u0026ndash;151 g). At this size, from the same genus, \u003cem\u003eS. chloronotus\u003c/em\u003e, the gonads have formed (Hoareau and Conand \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2001\u003c/span\u003e), so this size is categorized as potential broodstock and used for sex determination.\u003c/p\u003e \u003cp\u003eSex determination was conducted for 121 potential broodstock by injecting 0.2 \u0026micro;L of 0.5 M Potassium chloride solution (CAS 7447-40-7 Merck). Sixty-five were found as male with a length of 14.4\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7 cm and a weight of 77.8\u0026thinsp;\u0026plusmn;\u0026thinsp;22.4 g, and 56 were female with a length of 14.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7 cm and a weight of 88.7\u0026thinsp;\u0026plusmn;\u0026thinsp;28.2 g. Seventeen unknown sex with a length of 13.38\u0026thinsp;\u0026plusmn;\u0026thinsp;2.3 cm and a weight of 45.7\u0026thinsp;\u0026plusmn;\u0026thinsp;20.9 g were not used for further cultivation. After the sex was determined, the male and female were kept separately in two 2x1x1 m\u003csup\u003e3\u003c/sup\u003e concrete tanks, fed with fresh grounded \u003cem\u003eSargassum\u003c/em\u003e sp. and \u003cem\u003eUlva\u003c/em\u003e sp., in a 2:1 ratio, 3% of the total sea cucumber biomass for the regeneration of internal organs and gonad maturation. In addition to the two feeds, the sea cucumbers eat benthos that grow on the walls and bottom of the tank. After 11 months of culture, they have reached a length of 17.9\u0026thinsp;\u0026plusmn;\u0026thinsp;5.0 cm and a weight of 156.7\u0026thinsp;\u0026plusmn;\u0026thinsp;52.2 g for males and 17.7\u0026thinsp;\u0026plusmn;\u0026thinsp;5.2 cm and 137.7\u0026thinsp;\u0026plusmn;\u0026thinsp;27.5 g for females, and are ready to be used as test animals in this study.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eSpawning of cultivated broodstock\u003c/h3\u003e\n\u003cp\u003eBefore spawning, the F1 broodstock was measured, weighed, and divided into three size groups, namely small (S), medium (M), and large (L). The average length and weight of the male broodstock from the small size group (13.18\u0026thinsp;\u0026plusmn;\u0026thinsp;2.03 cm; 116.22\u0026thinsp;\u0026plusmn;\u0026thinsp;9.81g), medium (15.0\u0026thinsp;\u0026plusmn;\u0026thinsp;2.15 cm; 134.08\u0026thinsp;\u0026plusmn;\u0026thinsp;7.48 g), and large (16.46\u0026thinsp;\u0026plusmn;\u0026thinsp;3.28 cm; 152.56\u0026thinsp;\u0026plusmn;\u0026thinsp;19.01 g) groups. The average length and weight of the female broodstock for the small size group (12.40\u0026thinsp;\u0026plusmn;\u0026thinsp;1.14 cm; 109.80\u0026thinsp;\u0026plusmn;\u0026thinsp;5.72 g), medium (14.20\u0026thinsp;\u0026plusmn;\u0026thinsp;1.25 cm; 136.10\u0026thinsp;\u0026plusmn;\u0026thinsp;12.78 g), and large (15.90\u0026thinsp;\u0026plusmn;\u0026thinsp;2.56 cm; 170.20\u0026thinsp;\u0026plusmn;\u0026thinsp;7.92 g) groups. After grouping small, medium, and large broodstock categories, five pairs were found to represent each group. According to their group, each female was placed in a polycarbonate container with a volume of 30 L. Then, the male was placed in a plastic basket and put into the container to prevent the male and female from mixing (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Each size group had five replicates, so the number of spawners was 15 pairs. Spawning is carried out for two consecutive days during the new moon period.\u003c/p\u003e \u003cp\u003eThe seawater used for spawning passes through a sand filter and a 5 \u0026micro;m cartridge filter (Purerite PS-05, KEMFLO). After filtration, the seawater is stored in a concrete tank, sterilized using two 35-watt UV lamps (UV Lamp SAKKAI PRO 35 WATT) for 16 hours, and is ready to transfer into the spawning tank. Each spawning tank is covered with black plastic to maintain a stable temperature. The spawning is carried out for 2 days.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\n\u003ch3\u003eLarvae rearing\u003c/h3\u003e\n\u003cp\u003eEggs produced from each pair of broodstock size groups are harvested and incubated for 24 hours in 15 units of 100 L black plastic tanks according to treatment. After hatching, the larvae are stocked at a density of 200 individuals L\u003csup\u003e\u0026minus;\u003c/sup\u003e1 in 15 units of an 80 L larval rearing tank. Larvae were fed with \u003cem\u003eChaetoceros muelleri, Isochrysis galbana, Nitzchia\u003c/em\u003e sp., and \u003cem\u003eNavicula\u003c/em\u003e sp. according to their development (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eFeeding scheme for \u003cem\u003eStichopus horrens\u003c/em\u003e larval rearing until they reach the juvenile stage\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003eDay\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1\u0026ndash;6\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7\u0026ndash;12\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e13\u0026ndash;21\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e22\u0026ndash;45\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eChaetoceros muelleri\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c5\" namest=\"c2\"\u003e \u003cp\u003e10\u0026ndash;20 x 10\u003csup\u003e4\u003c/sup\u003e cell ml\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eIsochrysis galbana\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.5\u0026ndash;10 x 10\u003csup\u003e4\u003c/sup\u003e cell ml\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eNitzchia\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colspan=\"2\" nameend=\"c5\" namest=\"c4\"\u003e \u003cp\u003e0.5\u0026ndash;15 x 10\u003csup\u003e4\u003c/sup\u003e cell ml\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eNavicula\u003c/em\u003e sp.\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0.1\u0026ndash;1.75 x 10\u003csup\u003e4\u003c/sup\u003e cell ml\u003csup\u003e-1\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eWater replacement starts on the fifth day by slowly lowering the water by 20% and then adding UV-sterilized water using a plankton net. The water is changed every two days. Water quality monitoring, including temperature, is carried out daily, while pH, dissolved oxygen, ammonia, and nitrite are monitored twice a week. After 40 days of rearing, larvae metamorphosed into juveniles, ready to harvest, and were kept in a nursery tank.\u003c/p\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003eData Analysis\u003c/h2\u003e \u003cp\u003eThe variables observed include the length and weight of the broodstock, the number of broodstock spawning, the number of eggs, the egg diameter, the hatching rate, the development of the larvae, the survival rate, and the proportion of large, medium, and small juveniles at the end of the study. Egg diameter and total length of larvae were measured using a microscope connected to a camera (Nikon DXM 1200F) and Win-Roof ver. 5.0 software.\u003c/p\u003e \u003cp\u003eThe normality and homogeneity of all data sets were checked through the Non-Parametric One-Sample Kolmogorov-Smirnov and Levene Test, respectively (SPSS v. 20, IBM, Armonk, NY, USA). Differences among treatments were detected in One-Way ANOVA, followed by post hoc Tukey HSD. Data are expressed as mean SE of the mean (n\u0026thinsp;=\u0026thinsp;10\u0026ndash;15). The significance level was 0.05.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eSpawning of F1 broodstocks, fecundity, egg diameter, and hatching rate\u003c/h2\u003e \u003cp\u003eIn two days of spawning trials, three pairs of broodstock spawned in each small, medium, and large size group with an average number of eggs: 489,600\u0026thinsp;\u0026plusmn;\u0026thinsp;202,413, 420,000\u0026thinsp;\u0026plusmn;\u0026thinsp;217,607, and 684,800\u0026thinsp;\u0026plusmn;\u0026thinsp;215,556 eggs, respectively. Statistically, the number of eggs from the three broodstock size groups was not significantly different (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05) (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe egg diameters in all size groups differed significantly (F (2,6)\u0026thinsp;=\u0026thinsp;22.149, P\u0026thinsp;=\u0026thinsp;0.002), with the highest average found in the large size group (200.92\u0026thinsp;\u0026plusmn;\u0026thinsp;18.18 \u0026micro;m), followed by the medium and small size groups of 174.22\u0026thinsp;\u0026plusmn;\u0026thinsp;15.61 and 137.65\u0026thinsp;\u0026plusmn;\u0026thinsp;12.47 \u0026micro;m, respectively. Meanwhile, the average hatching rate in the large size group (82.12\u0026thinsp;\u0026plusmn;\u0026thinsp;5.37%) was not significantly different (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05) from the medium size group (67.84\u0026thinsp;\u0026plusmn;\u0026thinsp;7.64) but significantly different from the small size group (26.29\u0026thinsp;\u0026plusmn;\u0026thinsp;8.52) (F (2,6)\u0026thinsp;=\u0026thinsp;47.350, P\u0026thinsp;=\u0026thinsp;0.000).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eReproductive performance (number of eggs, egg diameter, and hatching rate) of F1 broodstock \u003cem\u003eStichopus horrens\u003c/em\u003e in different size groups\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSize Groups\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eBody weight of broodstock (g)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNumber of eggs (eggs/ind)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eEgg diameter (\u0026micro;m)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eHatching rate (%)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSmall\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e124.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e117.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e722,400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e132.61\u0026thinsp;\u0026plusmn;\u0026thinsp;8.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e34.56\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e105.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e105.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e355,200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e125.59\u0026thinsp;\u0026plusmn;\u0026thinsp;11.87\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e17.54\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e125.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e115.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e391,200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e154.76\u0026thinsp;\u0026plusmn;\u0026thinsp;17.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e26.78\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAverage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e118.40\u0026thinsp;\u0026plusmn;\u0026thinsp;9.50\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e112.40\u0026thinsp;\u0026plusmn;\u0026thinsp;5.10\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e489,600\u0026thinsp;\u0026plusmn;\u0026thinsp;202,413\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e137.65\u0026thinsp;\u0026plusmn;\u0026thinsp;12.47\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e26.29\u0026thinsp;\u0026plusmn;\u0026thinsp;6.96\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMedium\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e145.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e129.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e609,600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e163.77\u0026thinsp;\u0026plusmn;\u0026thinsp;11.58\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e79.40\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e130.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e125.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e182,400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e182.17\u0026thinsp;\u0026plusmn;\u0026thinsp;18.15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e69.82\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e127.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e158.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e468,000\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e176.73\u0026thinsp;\u0026plusmn;\u0026thinsp;17.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e84.30\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAverage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e134.30\u0026thinsp;\u0026plusmn;\u0026thinsp;7.70\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e137.50\u0026thinsp;\u0026plusmn;\u0026thinsp;14.57\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e420,000\u0026thinsp;\u0026plusmn;\u0026thinsp;217,607\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e174.22\u0026thinsp;\u0026plusmn;\u0026thinsp;15.61\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e77.84\u0026thinsp;\u0026plusmn;\u0026thinsp;6.01\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eLarge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e185.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e183.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e794,400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e184.05\u0026thinsp;\u0026plusmn;\u0026thinsp;16.19\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e85.26\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e150.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e166.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e436,800\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e201.40\u0026thinsp;\u0026plusmn;\u0026thinsp;19.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e75.92\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e141.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e162.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e823,200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e217.31\u0026thinsp;\u0026plusmn;\u0026thinsp;18.51\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e85.18\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAverage\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e159.20\u0026thinsp;\u0026plusmn;\u0026thinsp;19.00\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e170.33\u0026thinsp;\u0026plusmn;\u0026thinsp;9.10\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e684,800\u0026thinsp;\u0026plusmn;\u0026thinsp;215,556\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e200.92\u0026thinsp;\u0026plusmn;\u0026thinsp;18.18\u003csup\u003ec\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e82.12 \u0026plusmn; 4.38\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eDifferent alphabetic letters between the averages in the same column indicate statistically significant (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eLarval development, survival rate, and percentage of juveniles\u003c/h3\u003e\n\u003cp\u003eLarval development was not different in broodstock of different sizes. Early auricularia is characterized by a well-visible functional mouth, esophagus, cloaca, and ciliated band used for feeding and movement. Final auricularia is reached 13\u0026ndash;15 days after hatching, and on the 17th \u0026minus;\u0026thinsp;21st day, the larvae reach the doliolaria stage. Approaching the metamorphosis to the pentactula, the behaviour changes from planktonic to benthic larvae, the ciliated band disappears, five tentacles are visible, and the larvae move to the bottom of the rearing tank, occurring after 21 days of rearing (Figs.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e and \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe survival of larvae 40 days post-fertilization was significantly different (F (2,6)\u0026thinsp;=\u0026thinsp;35.816, P\u0026thinsp;=\u0026thinsp;0.000) between large-size groups and medium and small broodstock groups. Meanwhile, the medium and small-size broodstock groups did not differ significantly (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). The survival rate pattern of larvae for each size broodstock group is shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e. The water quality parameters in the larval rearing medium, including temperature, pH, dissolved oxygen, ammonia, and nitrite, are presented in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e and are still feasible to support sea cucumber larvae rearing.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e\n\u003ch3\u003e\u003c/h3\u003e\n\u003cdiv class=\"Heading\"\u003e\u003c/div\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAverage water quality measurement results during larval rearing\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eParameters\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSmall-size group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMedium-size group\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLarge-size group\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTemperature (\u0026deg;C)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Taba\" border=\"1\"\u003e \u003ccolgroup cols=\"1\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.09\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabb\" border=\"1\"\u003e \u003ccolgroup cols=\"1\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29.03\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"No\" id=\"Tabc\" border=\"1\"\u003e \u003ccolgroup cols=\"1\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e29.13\u0026thinsp;\u0026plusmn;\u0026thinsp;0.11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003epH\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e8.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.26\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDissolved oxygen (ppm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e5.30\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.25\u0026thinsp;\u0026plusmn;\u0026thinsp;0.07\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.08\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAmmonia (ppm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.12\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.10\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.02\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNitrite (ppm)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0.09\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e0.08\u0026thinsp;\u0026plusmn;\u0026thinsp;0.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e0.06\u0026thinsp;\u0026plusmn;\u0026thinsp;0.01\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eFigures \u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e and \u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e show the average length and weight of juvenile \u003cem\u003eS. horrens\u003c/em\u003e and the percentage of juvenile size groups. The analysis results show that each size group's body length, weight, and percentage are not significantly different (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e"},{"header":"Discussion","content":"\u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eSpawning of F1 broodstock, fecundity, egg diameter, and hatching rate\u003c/h2\u003e \u003cp\u003eTwo-year-old \u003cem\u003eS. horrens\u003c/em\u003e broods have succeeded in spawning for the three size groups studied, with a range of sizes; even small female and male broodstock groups with an average body weight of 105\u0026ndash;125 g can spawn, as well as broodstock in large and medium groups. The broodstock used in this research is much larger than other species. The first gonad maturity in \u003cem\u003eActinopyga echinites\u003c/em\u003e was detected at a total weight of 65 g (Kohler et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), and Sembiring et al. (\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) reported that F1 broodstock \u003cem\u003eH. scabra\u003c/em\u003e can spawn at an average body weight of 122.6\u0026thinsp;\u0026plusmn;\u0026thinsp;32.37 g. This indicates that the nutritional reserves of the broodstock play an important role and can improve reproductive performance. The quantity and quality of these nutrient reserves determine the fecundity and quality of the eggs, as also mentioned by Schreck et al. (\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Thus, the energy source required for embryo and larval development depends mainly on the broodstock's nutrient reserves (Huang et al. \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe three-size broodstock groups show the same spawning frequency, and the number of eggs produced did not differ significantly. These results show that the weight of the brooder does not always affect the number of eggs produced, especially because there is a large difference in the number of eggs between individuals in one brood size group. Given that the broodstock used in this experiment is the same age, the maintenance from seed to broodstock is done under the same environmental and feed conditions. Thus, it can be concluded that if broodstock rearing is optimal, then size is not the main factor in the success of spawning. This is in line with Gianasi (\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2018\u003c/span\u003e), who reports that in \u003cem\u003eCucumaria frondosa\u003c/em\u003e, optimal environmental conditions can increase egg reproduction in addition to the broodstock's size. Furthermore, apart from environmental conditions, other external factors that can affect broodstock productivity include the type of feed given, the maintenance container, the phase of the moon, and the season (photoperiod and temperature) \u003cb\u003e(\u003c/b\u003eHu et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Laguerre et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Rakaj et al. \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Tehranifard et al. 2007). Ru et al. (\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) also reported that sea cucumbers use nutritional sources to support gonad growth, so their reproductive performance depends on the availability and quality of food. The feed given to the broodstock during maintenance is ground \u003cem\u003eSargassum\u003c/em\u003e sp. and \u003cem\u003eUlva\u003c/em\u003e sp. In addition, the broodstock also eats benthos that grow naturally in the broodstock maintenance tank. Moreover, Wen et al. (\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) reported that \u003cem\u003eA. japonicus\u003c/em\u003e and \u003cem\u003eS. horrens\u003c/em\u003e prefer \u003cem\u003eSargassum\u003c/em\u003e sp. as food. Xu et al. (\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) also reported that the food source for \u003cem\u003eStichopus monotuberculatus\u003c/em\u003e comes from \u003cem\u003eSargassum\u003c/em\u003e sp., phytoplankton, and organic matter.\u003c/p\u003e \u003cp\u003eThe average diameter of eggs from the three size groups of F1 broodstock \u003cem\u003eS. horrens\u003c/em\u003e was significantly different (\u003cem\u003eP\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/em\u003e), with the highest average egg diameter (217.31\u0026thinsp;\u0026plusmn;\u0026thinsp;18.51 \u0026micro;m) obtained from the large size group broodstock. Conand (\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e1993\u003c/span\u003e) reported that the \u003cem\u003eStichopus variegatus\u003c/em\u003e begins maturing at a length of 270 mm and with an oocyte diameter of 180 \u0026micro;m, but its fecundity is only 10,000 oocytes. The egg diameter in this experiment is far bigger, even compared to the small-size group (132\u0026ndash;154 \u0026micro;m). De La Rosa et al. (\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2023\u003c/span\u003e) \u003cb\u003es\u003c/b\u003etate that the egg's large diameter indicates a more significant food reserve that can support the development of the embryo and larvae when hatching, making these conditions excellent for the survival of sea cucumber larvae.\u003c/p\u003e \u003cp\u003eThe egg-hatching rates of the large and medium groups differed significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) from the small groups. The highest hatching rate is obtained from the large group, followed by the medium-sized group, so the lowest hatching rate is in the small group. The condition of the female broodstock can affect egg quality and hatching rate. Marquet et al. (\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) reported that the egg-hatching rates of \u003cem\u003eHolothuria arguinensis\u003c/em\u003e and \u003cem\u003eH. Mamata\u003c/em\u003e were influenced by egg quality, fertility rates, and environmental conditions. The same has been reported in sea urchins \u003cem\u003eParacentrotus lividus\u003c/em\u003e (Guettaf et al. \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2000\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eDuring spawning and egg incubation, salinity and temperature were maintained at 33\u0026ndash;34 ppt and 30\u0026ndash;31\u0026deg;C for all broodstock size groups, and these ranges are optimum for most sea cucumbers, as reported by Hu et al. (\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). The salinity level is one of the significant environmental factors affecting the hatching rate. Tu et al. (\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) reported that the Japanese sea cucumber \u003cem\u003eApostichopus japonicus\u003c/em\u003e can produce a high hatching rate at a salinity of 34 ppt. Asha et al. (\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) also noted that the optimal salinity range for hatching \u003cem\u003eH. scabra\u003c/em\u003e eggs and rearing the larvae to the auricularia stage is 33\u0026ndash;35 ppt. Furthermore, Falconer and Mackay (\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1996\u003c/span\u003e) also state that the phenotype of an organism is influenced by its genotype, environment, and interaction. In this study, large broodstock groups resulted in higher hatching rates. This suggests that the size of sea cucumber broodstock affects the hatching rate of eggs, and it can be assumed that this hatching rate is primarily an inherited trait.\u003c/p\u003e \u003cp\u003eCompared to the performance of the natural broodstock, which has a high egg production of 1\u0026ndash;2\u0026nbsp;million eggs, the egg production of the F1 broodstock is still much lower, at 400,000-600,000 eggs. Environmental conditions in nature, which vary the availability of food, result in a higher fecundity of natural broodstock than domesticated broodstock, which is kept in tanks and fed a limited type of food. This difference in fecundity is likely related to higher nutrition due to the availability of more varied foods during broodstock rearing in ponds. During rearing in the concrete tank for the regeneration of the gastrointestinal tract and gonads, gonad maturation, and spawning of the broodstock, the feed given is limited to \u003cem\u003eSargassum\u003c/em\u003e sp., \u003cem\u003eUlva\u003c/em\u003e sp., and benthos that grow on the inner wall of the tank. Therefore, research on natural and artificial feeds for domesticated broodstock still needs to be continued. Cong et al. (\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2024\u003c/span\u003e) state that the variation in fecundity in sea cucumbers is related to the difference in the amount of food consumed by sea cucumbers.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003eLarval development, survival rate, and percentage of juveniles\u003c/h2\u003e \u003cp\u003eThe development of sea cucumber larvae from different broodstock size groups did not differ significantly (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). Larvae of \u003cem\u003eS. horrens\u003c/em\u003e of large broodstock size have a high survival rate and significantly differ from medium and small broodstock sizes. In addition, larger sizes have more energy sources to provide nutrients for the development of embryos and larvae when hatching, making this condition very favourable for the survival of larvae (Widyastuti et al. \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). This can be seen from the diameter of the eggs produced by large-sized broodstock, which is more significant than the diameter of the eggs in other broodstock size groups. Morgan (\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) reports that \u003cem\u003eAustralostichopus mollis\u003c/em\u003e larvae from larger-sized broodstock tend to have better access to food sources in the early developmental phase, which supports increased growth and larval survival. Peters-Didier and Sewell (\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2008\u003c/span\u003e) reported a relationship between egg size and lipid content, as the lipid content in eggs is a significant energy reserve in the early development of \u003cem\u003eAustralostichopus mollis\u003c/em\u003e larvae. This makes it possible for larger broodstock sizes to provide more energy to each egg, which in turn has the potential to produce better larvae.\u003c/p\u003e \u003cp\u003eSome research results state that the larval development phase of the \u003cem\u003eS. horrens\u003c/em\u003e sea cucumber is the same as that of other \u003cem\u003eAspidochirotes\u003c/em\u003e sea cucumbers (Chen and Chian \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Hu et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; McEuen \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1988\u003c/span\u003e; Ramofafia et al. \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2003\u003c/span\u003e). The food and environmental factors during larval rearing significantly affect juvenile survival (Yu et al. \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Water quality is important in a sea cucumber hatchery because larvae are susceptible to environmental changes (Asha and Muthiah \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). This study's water temperature, pH, dissolved oxygen, ammonia, and nitrite were relatively consistent and generally within the normal range for maintaining sea cucumber larvae of tropical holothurian species (Hu et al. \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Knauer \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2011\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThe first generation of domesticated \u003cem\u003eS. horrens\u003c/em\u003e broodstock from the large broodstock group (170.33\u0026thinsp;\u0026plusmn;\u0026thinsp;9.10 g) provides better reproduction and larval performance.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAuthor contribution\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSari Budi Moria Sembiring and Ketut Maha Setiawati joined in all activities, from arranging the idea and designing the experiment, broodstock, larval and juvenile rearing, samples collection, larval measurement, statistical analysis, data interpretation, and preparation of the original draft. Jhon Harianto Hutapea, Gunawan Gunawan, Ananto Setiadi, and Ni Wayan Widya Astuti joined in conducting broodstock, larval and juvenile rearing, sample collection, larval measurement, and statistical analysis. Haryanti Haryanti and Nyoman Adiasmara Giri arranged the idea, designed the experiment, performed data interpretations, and prepared the original draft. Rarastoeti Pratiwi performed data interpretations and prepared the original draft. All authors have read and agreed to the published version of the manuscript\u003cstrong\u003e.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe Ethics Commission for Animal Care and Use of the Indonesian National Research and Innovation Agency decided that this study does not require Ethics Clearance, and the animals in this study are invertebrates that are exempt from Ethics Clearance (Reference Number: 215/KE.02/SK/12/2023).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research was funded by the Directorate of Research and Innovation Funding, Deputy for Facilitation of Research and Innovation, National Research and Innovation Agency (No. 61/II.7/HK/2024).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interest\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflicts of interest regarding this research article.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgments\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are grateful to all technicians at the sea cucumber hatchery for their technical assistance during the experiment and to the Directorate of Research and Innovation Funding, Deputy for Facilitation of Research and Innovation, National Research and Innovation Agency.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAsha, P. 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Article ID\u003c/em\u003e. 8947997. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1155/2022/8947997\u003c/span\u003e\u003cspan address=\"10.1155/2022/8947997\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2022).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true},"keywords":"broodstock size, cultivation, performance, reproduction, Stichopus horrens","lastPublishedDoi":"10.21203/rs.3.rs-6712637/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6712637/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe production of cultivated sea cucumber \u003cem\u003eStichopus horrens\u003c/em\u003e broodstock is urgently being carried out, considering the sharp decline of natural broodstock. Therefore, natural broodstock (F0) were spawned, and their larvae were reared in a hatchery until they metamorphosed into seeds (F1). Further cultured in the earthen pond for 16 months to reach broodstock size. After determining the sex by the eviceration method, males and females were cultured in separate concrete tanks measuring 2x1x1m\u003csup\u003e3\u003c/sup\u003e to regenerate internal organs and gonad maturation, and fed with fresh grounded \u003cem\u003eSargassum\u003c/em\u003e sp. and \u003cem\u003eUlva\u003c/em\u003e sp. at 3% biomass. Eleven months later, the F1 broodstock was divided into three groups based on body weight: small (116.22\u0026thinsp;\u0026plusmn;\u0026thinsp;9.81 g and 109.80\u0026thinsp;\u0026plusmn;\u0026thinsp;5.72 g), medium (134.08\u0026thinsp;\u0026plusmn;\u0026thinsp;7.48 g and 136.10\u0026thinsp;\u0026plusmn;\u0026thinsp;12.78 g), and large (152.56\u0026thinsp;\u0026plusmn;\u0026thinsp;190.01 g and 170.20\u0026thinsp;\u0026plusmn;\u0026thinsp;7.92 g) for male and female, respectively. Five pairs from each broodstock group were spawned separately in fifteen 30 L polycarbonate tanks. The F1 broodstock could produce 180,000 to 820,000 eggs per broodstock, but not significantly different (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05) among broodstock size groups. The highest egg diameter (200.92\u0026thinsp;\u0026plusmn;\u0026thinsp;18.18 \u0026micro;m) and larval survival were obtained in the large broodstock size group and were significantly different (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) from the others. The highest hatching rate (82.12\u0026thinsp;\u0026plusmn;\u0026thinsp;4.38%) was obtained in large broodstock size, followed by medium broodstock size, and was significantly different (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05) from the small broodstock size group. Mean F1 is suitable for domesticated broodstock, especially from large broodstock size groups.\u003c/p\u003e","manuscriptTitle":"Reproductive Performance of the First Generation Stichopus horrens Broodstock","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-30 18:31:30","doi":"10.21203/rs.3.rs-6712637/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-08-12T08:34:21+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-07-06T11:04:51+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"104393354572102969500470676064546106517","date":"2025-06-09T04:22:37+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-05-28T00:55:44+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-05-28T00:53:01+00:00","index":"","fulltext":""},{"type":"editorInvited","content":"","date":"2025-05-27T12:57:02+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-05-23T06:57:58+00:00","index":"","fulltext":""},{"type":"submitted","content":"Scientific Reports","date":"2025-05-23T06:56:50+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"scientific-reports","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"scirep","sideBox":"Learn more about [Scientific Reports](http://www.nature.com/srep/)","snPcode":"","submissionUrl":"","title":"Scientific Reports","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"stoa","reportingPortfolio":"Scientific Reports","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"1437d3f3-2337-4107-9912-fb8cf316bc97","owner":[],"postedDate":"May 30th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":49143213,"name":"Biological sciences/Biological techniques"},{"id":49143214,"name":"Biological sciences/Developmental biology"},{"id":49143215,"name":"Biological sciences/Physiology"}],"tags":[],"updatedAt":"2025-12-22T15:59:00+00:00","versionOfRecord":{"articleIdentity":"rs-6712637","link":"https://doi.org/10.1038/s41598-025-30858-w","journal":{"identity":"scientific-reports","isVorOnly":false,"title":"Scientific Reports"},"publishedOn":"2025-12-18 15:56:56","publishedOnDateReadable":"December 18th, 2025"},"versionCreatedAt":"2025-05-30 18:31:30","video":"","vorDoi":"10.1038/s41598-025-30858-w","vorDoiUrl":"https://doi.org/10.1038/s41598-025-30858-w","workflowStages":[]},"version":"v1","identity":"rs-6712637","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6712637","identity":"rs-6712637","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}