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Hemibagrus wyckii, endangered species, alternative fish species, egg quality, sperm
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Aryani N, Suharman I and Syandri H. Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.12688/f1000research.14746.1)
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Research Article
Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture
[version 1; peer review: 2 approved]
PUBLISHED 31 May 2018
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Abstract
Corresponding author:
Netti Aryani
Competing interests:
No competing interests were disclosed.
Grant information:
This study was funded by a study grant (Riset Dasar Unggulan Perguruan Tinggi) from the Directorate of Research and Community Service, Ministry of Research Technology and Higher Education Republic of Indonesia (No. 311/UN.19.5.1.3/PP/2018)
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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© 2018 Aryani N et al.
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Data associated with the article are available under the terms of the Creative Commons Zero "No rights reserved" data waiver (CC0 1.0 Public domain dedication).
How to cite: Aryani N, Suharman I and Syandri H. Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.12688/f1000research.14746.1)
First published: 31 May 2018, 7:683 (https://doi.org/10.12688/f1000research.14746.1)
Latest published: 03 Sep 2018, 7:683 (https://doi.org/10.12688/f1000research.14746.2)
The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
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Introduction
In Indonesia, the fisheries sector plays an important economic role through income generation, diversification of livelihoods, supply of animal proteins, and foreign exchange earnings1. During recent decades, in the freshwater aquaculture sector, the prioritized species for culture were Clarias, Pangasius, Tilapia, Common carp and Giant gourami. Wild fish species in rivers, reservoirs and lakes have not been prioritized for aquaculture operations.
In the Riau Province, there are three rivers, the Kampar Kanan, Kampar Kiri, and Siak rivers, and the Koto Panjang Reservoir. The Kampar Kanan river hosts up to 34 fish species2, the Kampar Kiri river hosts up to 86 fish species3, the Ukai river, a branch of Siak River hosts up to 31 fish species4 and Koto Panjang Reservoir hosts up to 26 fish species5.
Hemibagrus wyckii (Bagridae) is one of the most important economic fish species that lives in the rivers and reservoir in Riau Province. H.wyckii (its local name is geso) is a carnivorous freshwater finfish native to Indonesia2,6. H. wyckii has been categorized as of “least concern” by the International Union for Conservation of Nature (IUCN). However, H.wyckii in the Kampar Kanan river was categorized as a vulnerable to endangered species7,8.
Due to the endangered population of H. wyckii, it is necessary to domesticate this species as an aquaculture candidate in the future. Therefore, the present study aimed to determine the reproductive performance of H. wyckii as a potential species under culture conditions to provide preliminary scientific information and evaluation.
Methods
Ethical considerations
While H. wyckii is classified as vunerable to endangered in the Kampar Kanan river, the Government of the Republic of Indonesia does not require licences to be obtain to capture and rear this species, hense no licences are applicable to this study. No animals suffered as a result of the activities of this study. H. wyckii was transported to the pond farm for rearing, injection, ovulation, stripping and sperm production. In the end of the experiment the H. wyckii still in good condition until return back to the pond.
Rearing and selection of breeders
Broodfish of H. wyckii were collected from upstream areas of the Kampar Kanan river in the Kouk village (0° 19' 23.44˝ N and 100° 56' 40.05˝ E), Kampar Regency, Riau Province. The broodfish kept in oxygenated polythene bag and transported by truck to Sadarlis Green Catfish Farm, Sungai Paku, Kampar Regency, Indonesia. Then, the broodstock of H. wyckii had been adapted and grown to maturation under the farm conditions. Prior to stocking, female and male fish were weighed using balance scale (OHAUS model CT 6000-USA), and their lengths were measured using a meter ruler with 0.01mm accuracy. During the grow-out period, fish were cultured in two ponds (4 x 4 x 3 m) separated by sex. The depth of water in each pond was 2.0 m. The inlet of the pond water come from Sungai Paku reservoir at a rate of 2.0 m3 per sec. The broodfish fed with freshwater seashell meat (Pilsbryoconcha exilis; Unionidae) collected from local fisherman near to Sungai Paku Reservoir. The seashell meat was kept at cold box with the temperature 5°C prior given to broodfish. According to Aryani et al.6 the proximate composition (% wet weight base) of the seashell meat was 89.37% moisture, 7.08% crude protein, 0.82% fat, 0.29% crude ash and 2.44% carbohydrate. The total of seashell meat given to broodfish everyday were 2.500g per pond (equivalent with 9% body weight of population). The feeding time at 17:00 pm due to carnivorous broodfish. The average weight and length from ten (10) of the female broodfish were 2,669.4±486.917 g and 62.84±8.20 cm, respectively. Meanwhile, the ten (10) of male broodfish were 1,769.1±401.10 g and 54.52±7.17 cm, respectively.
The fish were checked monthly for ovulation and semen production from mid November, 2017 onwards. The broodfish were captured with a gillnet formed into a net bag with the appropriate mesh size and anesthetized orally with Tricaine methanesulfonate (MS-222, ethyl 4-aminobenzoate methanesulfonate 98%, Sigma Aldrich Co, USA, MO; 50 mg L-1), based on the dosage used for Solea senegalensis9. Oocyte maturation was assessed for each individual. The fish were returned to their pond after evaluation, and no mortality occurred. Fish were fasted 48 h prior to the evaluation. Oocytes sampled in vivo were taken from females using the method described by Nowosad et al.10 and were placed in Serra's solution (6:3:1 of 70% ethanol, 40% formaldehyde and 99.5% acetic acid) for clarification of the cytoplasm. After 5 min, the position of the stage IV oocyte nucleus was determined using criteria by Krejszeff et al.11 and was classified as germinal vesicle in the periphery or germinal vesicle breakdown (GVBD).
Female reproductive performance
H. wyckii was categorized as endangered species and difficult to obtain in Kampar Kanan river. A total of 10 mature have been eligible for the experiment. 10 mature females that had oocyte stage IV were sampled from broodstock from 3rd week of February to March 2018 in the same farm, and live weights (FeW) and total lengths (FeL) were measured after anesthetization with 0.50 mg L-1 MS-2229. For ovulation, each female broodfish received two injections of GnRH analogs with a dopamine antagonist (Ovaprim) (manufactured for Syndel Laboratories Ltd, 2595 McCullough Rd. Nanaimo, B.C.V9S 4M9 Canada) applied intraperitoneally under the left pectoral fin. The first injection was 0.2 mL kg BW-1 and the second was 0.6 mL kg BW-1 (total 0.8 mL kg BW-1) at 12 h intervals. These dosages refer to the previous doses for ovulation of H. wyckii6. At 18 to 20 h after injection, eggs were stripped into a plastic vessel. Eggs were fertilized using the “dry method” as described by Dabrowski et al.12 Egg weights of each female were by determined weighing 50 eggs to the nearest 0.01 g, and egg diameters were measured to the nearest 0.01 mm. Egg size (50 eggs for each fish) was measured using an Olympus microscope. Then, a balanced saline solution (7.5 g of NaCl, 0.2 g of KCl, 0.2 g of CaCl22H2O, and 0.02 g of NaHCO3, in 1000 mL distilled water) was added over the eggs13, followed by an addition of pooled sperm from 10 males. The eggs were then gently mixed for fertilization and left for three minutes. The fertilized eggs were rinsed several times with incubation water to remove sperm remnants as well as dead and broken eggs. The eggs were left for an additional 25 minutes to facilitate egg hardening by water absorption and disinfected with 100 ppm iodine for 10 minutes. Then, eggs were transferred to incubation trays placed in a vertical hatching system. The water flow rate to each vertical incubator was 3 L minˉ¹. Fifty eggs were randomly sampled at 15 h after fertilization to determine the fertilization rate (FR). The hatching rate (HR) was determined by counting all hatched fry.
Determination of sperm quality
Males were stimulated with a half-dose of the same hormonal preparations used to stimulate the females. Semen samples were obtained from 10 fish randomly selected from the farm. The male fish were anesthetized with 50 mg Lˉ¹ of MS-222. The doses of the anaesthetic agents were prepared a few minutes before each experiment based on the methods of Weber et al.9, and then, weights (MaW) and total lengths (MaL) were measured. Special care was taken to avoid any contamination of semen with urine, feces, mucus and water. Semen samples were collected using plastic syringes in 3 mL aliquots, then placed in an insulated ice-cooled container, transported to the laboratory and analyzed within 2 h.
The sperm evaluation included gross (visual) and microscopic examination (as reviewed by Rurangwa et al.14, and Cabrita et al.15. The gross examination was based on visual and physical observation of parameters such as the semen volume by collecting the semen in a graduated cylinder and determining the level in milliliters. The microscopic examination was carried out using an Olympus model CX40, with magnification between X 10 and X 25 to determine other parameters such as: motility (duration and percentage). Motility (MO) percentage and duration were determined by observing water activated semen placed on a glass slide under a microscope. Motile sperm were observed and expressed as a percent of non-moving sperm. Motility duration (DMO) was determined as the period between movements of the sperm to cessation of any progressive movement expressed in seconds. Sperm concentration (SC) was measured under a microscope using Neaubeaur’s hemocytometer and calculated as the number of sperm mlˉ¹16. Semen pH was determined with a hand pH meter (HI8424 Hanna Instruments, USA).
Water quality
The water temperature of the farm was measured with a thermometer (Celsius scale), and water samples were collected to determine the dissolved oxygen (DO) concentrations. An oxygen meter (YSI model 52, Yellow Spring Instrument Co., Yellow Springs, OH, USA) was used in situ, and pH values were determined with a pH meter (Digital Mini-pH Meter, 0-14PH, IQ Scientific, Chemo-science Thailand Co., Ltd, Thailand). Alkalinity and hardness levels of the water were measured in each replicate according to standard procedures17. The water quality parameters were measured once per month.
Statistical analysis
Results were given as the means ± SD. Simple linear regression analyses were performed using SPSS software (version 16.0 for Windows; SPSS Inc., Chicago, IL). The standard deviation of each parameter was determined. For linear regression analysis, significant correlations were considered at p<0.05.
Results
Descriptive measurements and the reproductive performance of female H.wyckii are presented in Table 1. Fifty percent of eggs hatched at 60 h (29–30 °C water temperature). The fertilization rate varied between 53.2 and 68.3%, whereas the hatching rate was between 39.5 and 48.3%.
Table 1. Female size, egg characteristics and hatchery performance of Hemibagrus wyckii (Mean ± SD).
Characteristics of male fish and sperm samples are presented in Table 2. The average live weight of the males is 1,769.1±401.1g. Male H.wyckii are found to be slightly smaller than females. In the genital maturation stage, the papilla is not prominent for all male fish as the second sexual characteristic of the other Hemibagrus.
Table 2. Male size and sperm characteristics of Hemibagrus wyckii (Mean ± SD).
According to the analysis of the linear relationship (r2) between variables of H. wyckii females shown in Table 3, there was a strong linear relationship between AF and FeW, AF and RF, HEW and EW, HW and EW, HW and HEW, and HR and FR. In contrast, the analysis of the linear relationship (r2) between variables of H. wyckii males shown in Table 4, show a strong linear relationship between MaW and MaL, MaW and GW, and MaL and GW.
Table 3. Correlations of variables (r2) for Hemibagrus wyckii females.
Statistically important at r2 > 0.500 (underlined)
Few: Female fish weight, FeL: Female fish length, AF: Absolute fecundity, RF: Relative fecundity, EW: Egg weight, HEW: Hardened egg weight, EWI: Eggs weight increase, ED: Egg diameter, HED: Hardened egg diameter, HDI: Hardened egg diameter increase, FR: Fertilization rate, HR: Hatching rate, HW: Hatching weight, GI: Gonadosomatic index.
Table 4. Correlations of variables (r2) for Hemibagrus wyckii males.
The temperature of the pond water ranged from 28°C to 29°C, oxygen ranged from 6.5 mg L-1 to 6.7 mg L-1, pH ranged from 6.5 to 6.8, alkalinity ranged from 42.97 mg L-1 to 57.33 mg L-1 and hardness ranged from 104.83 mg L-1 to 110.51 mg L-1.
Dataset 1.Data of female size, egg characteristic and hatchery performance of Hemibagrus wyckii.
Discussion
In our study, the spawning period of H. wyckii started at the 3rd week of February and continued until the final examination in March. However, our monthly observations of H.wyckii captured by fishermen in the Kampar Kanan river found fish at the gonadal I, II, III and IV stages of development (stage scale by Krejszeff et al11), showing a spawning type shows of a partial spawner. Spawning in the wild occurs at the start and end of the rainy season, or this species could spawn twice per year. The spawning type of H. wyckii is the same as that of Hemibagrus nemurus18. The duration from fertilization to a 50% hatching rate was 60 h with 29°C to 30°C water temperature. The fertilization rate and hatching rate of various Hemibagrus species are reported in Table 5. When findings are compared with other Hemibagrus species, the duration of hatching is longer in H.wyckii. In other words, H.wyckii has species specific characteristics the regarding hatching rate, because the egg diameter was bigger than other Hemibagrus eggs (Table 5).
AFs of H.wyckii were between 4125 and 9958 eggs/fish and RFs were between 1400 and 3000 eggs kgˉ¹. Egg production per kg fish (RF) is thought to be more informative than absolute fecundity. RF values of H.wyckii were lower compared with those of H.nemurus19–21. In our study, there was no strong linear relationship between RF and fish size. Meanwhile, there was a strong linear relationship between AF and fish size (Table 3).
Table 5. Summary of the egg diameter, fertilization and hatching rate of H.wyckii, with respect to those reported for H.nemurus and H.dibrugarensis.
| Species | Eggs diameter (mm) | Fertilization rate (%) | Hatching rate (%) | Reference |
|---|---|---|---|---|
| H. wyckii | 2.10 – 2.86 | 53.2 – 68.3 | 39.5 – 58.3 | This study |
| H. wyckii | 2.79 – 2.85 | 45.8 – 60.2 | 37.33 – 55.1 | 6 |
| H. nemurus | 1.13 – 1.22 | 74.16 – 77.83 | 64.16 – 67.83 | 19 |
| H. nemurus | 1.07 – 1.21 | 70.50 – 81.4 | 63.5 – 71.4 | 20 |
| H.dibrugarensis | 34.83 – 77.54 | 20.61 – 74.32 | 24 | |
| H.nemurus | - | 93.5 – 94.5 | 56.91 – 78.3 | 25 |
| H. nemurus | 1.57 – 1.75 | 81.57 – 98.54 | 81.3 – 96.72 | 21 |
| H. nemurus | 39.5 – 79.0 | 32.5 – 69.7 | 18 |
EDs and EWs obtained here ranged between 2.10 and 2.86 mm and 24.4 and 31.8 mg respectively, consistent with those reported by Aryani et al.6 At the end of the hardening process, the increases in egg weight and diameter were 29.86 and 14.9% respectively. In the present study, there were strong linear relationships between EW and HEW, and between EW and HW (Table 3). Lahnsteiner and Patzner22 state that egg weight increases after the hardening process and is linearly correlated with the viability of eggs in Rainbow trout, but not in Alakir trout23.
In this study, the FRs of H. wyckii were higher than those of H.wyckii from research conducted by Aryani et al.6. This suggests that we have improved the method for the fertilization process of sperm and eggs so that hatching rate can increase. Nevertheless, the fertilization rate of H.wyckii was lower than that of H.nemurus17,24,25 (Table 5). When HR values are compared with other Hemibagrus species, HR levels are lower than those of Aryani and Suharman14, Adebiyi et al.25 and Suhenda et al.21 but higher than those of Aryani et al.6 There were strong positive correlations with FR and HR (r2 = 0.91) (Table 3). Meanwhile, FR and HR were not positively correlated with ED (Table 3).
Gonadotropins (GTHs), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and sex steroids are the key regulators of reproduction26–28. Moreover, numerous circulating endocrine and locally acting paracrine and autocrine factors regulate the various stages of oocyte development and maturation29,30. Other factors that significantly affect fish eggs are genetic, environmental and stress factors31–33. However, there is no information about the effects of such kinds of factors on the embryonic development of H.wyckii. There are also further information requirements concerning hatchery management of H.wyckii such as feed levels and feed type6. During the adaptation period in the present instance, H. wyckii were fed with meat freshwater seashell (Pilsbryoconcha exilis; Unionidae), the local name of which is “lokan” which may not be fully suitable for this species, even though this species is carnivorous. The requirement of a balanced feed that meets the nutritional requirements of species being cultured34–36 and application of a proper feeding program during the ovarian development37 have been emphasized. Suhenda et al.21 reported that diet should be offered to H. nemurus broodstock at an 8% lipid level in fish feed with 35% crude protein for 4 months to obtain high quality gametes. Meanwhile, Aryani and Suharman20 suggest that a minimum of 32% crude protein should be included in the diet of female H.nemurus broodstock. Additionally, implantation of 17ß-estradiol has also managed to improve the reproductive performance of H.nemurus19. These issues in H.wyckii are yet to be elucidated. Therefore, such research is very important for H.wyckii in the future.
Average SVs determined in H. wyckii are higher than those in H.nemurus (0.10 to 0.35 mL)18 and Clarias gariepinus38, but lower than those reported for Ictalurus punctatus39. It appears that the semen volume in other species has a positive relationship with sperm concentration, including in Ictalurus punctatus39. Meanwhile, in fish farms and hatcheries, the biotic and abiotic factors that affect sperm quality are diverse and dependent on complex interactions between genetic, physiological and environmental factors14. On the other hand, improvements in broodstock nutrition and feeding greatly improve gamete quality and larvae production40.
Semen pH values of H.wyckii are consistent with those of other species, including Barbus grypus41 and Carrasius gibelio42. The sperm motility of H.wyckii between 70.2 and 75.50%, and the duration of motility was between 40.0 and 54.0 sec, results that are consistent with Ictalurus punctatus39. According to Effer et al.43 the duration of sperm motility in fish depends on the temperature of the activation medium. Sperm of H.wyckii had an effective fertilizing ability according to the correlation analysis, which did not detect any significant relationship between FR and sperm parameters. However, there was a positive relationship between MO and DMO (r2 = 0.49). Sperm morphology, density, volume, motility and fertilizing capacity, as well as the composition and osmolality of the seminal plasma are parameters commonly measured to assess sperm quality in fish14,44. In this study, we did not investigate the ionic composition of the semen, but this phenomenon could be related to the ionic composition of semen, which has a significant influence on motility and duration of motility45–47.
Conclusion
In conclusion, the reproductive performance of H.wyckii under culture conditions is within the range of available data for other Hemibagrus species. Considering that successful larvae production (and potential juvenile growth) was possible using the same methods as in Bagridae. H.wyckii can be an alternative species for aquaculture. However, further studies are clearly required to determine several aspects of this fish under culture conditions.
Data availability
Dataset 1: Data of female size, egg characteristic and hatchery performance of Hemibagrus wyckii 10.5256/f1000research.14746.d20432848
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how to cite this article
Aryani N, Suharman I and Syandri H. Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.12688/f1000research.14746.1)
NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.
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Version 1
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PUBLISHED 31 May 2018
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How to cite this report:
Nugroho RA. Reviewer Report For: Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.5256/f1000research.16043.r37278)
The direct URL for this report is:
https://f1000research.com/articles/7-683/v1#referee-response-37278
https://f1000research.com/articles/7-683/v1#referee-response-37278
NOTE: it is important to ensure the information in square brackets after the title is included in this citation.
Reviewer Report 22 Aug 2018
Approved
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Title
The title should be revised: delete “a candidate species for aquaculture”. It needs more parameters (Growth, feed aspect, etc.) to convey that this species can be a candidate for aquaculture.
Abstract
In methods: ... Continue reading
The title should be revised: delete “a candidate species for aquaculture”. It needs more parameters (Growth, feed aspect, etc.) to convey that this species can be a candidate for aquaculture.
Abstract
In methods: ... Continue reading
CITE
HOW TO CITE THIS REPORT
Nugroho RA. Reviewer Report For: Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.5256/f1000research.16043.r37278)
The direct URL for this report is:
https://f1000research.com/articles/7-683/v1#referee-response-37278
https://f1000research.com/articles/7-683/v1#referee-response-37278
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
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22
How to cite this report:
Muchlisin ZA. Reviewer Report For: Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.5256/f1000research.16043.r34564)
The direct URL for this report is:
https://f1000research.com/articles/7-683/v1#referee-response-34564
https://f1000research.com/articles/7-683/v1#referee-response-34564
NOTE: it is important to ensure the information in square brackets after the title is included in this citation.
Reviewer Report 11 Jun 2018
Approved
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Title:
- I think the title should be revised as "Reproductive biology and breeding of the Asian catfish (Hemibagrus wyckii) after domestication process.
- In the title: (Hemibagris wyckii Bagridae) is missed
CITE
HOW TO CITE THIS REPORT
Muchlisin ZA. Reviewer Report For: Reproductive performance of asian catfish (Hemibagrus wyckii Bagridae), a candidate species for aquaculture [version 1; peer review: 2 approved]. F1000Research 2018, 7:683 (https://doi.org/10.5256/f1000research.16043.r34564)
The direct URL for this report is:
https://f1000research.com/articles/7-683/v1#referee-response-34564
https://f1000research.com/articles/7-683/v1#referee-response-34564
NOTE: it is important to ensure the information in square brackets after the title is included in all citations of this article.
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