鱼类生殖生态学研究的基本方法
Summary
描述了一系列基本方法来研究养殖在水族箱中的鱼类的生殖生态。这些是使用SCUBA收集鱼类，运送活鱼以及观察保持在水族箱中的野生捕捞鱼的繁殖行为的有用方案。
Abstract
当持续的实地调查是不可能的时候，养殖观察对于揭示鱼行为和生态学方面是有价值的。在这里，描述了一系列基本技术，以便观察野生捕获的小型鱼的生殖行为，作为模型，保存在水族箱中。该方法着重于三个步骤：收集，运输和观察底物spavner的生殖生态学。活鱼收集和运输的基本方面是（1）防止对鱼的伤害，（2）仔细适应水族馆。收集活鱼时，预防伤害如划痕或突然变化的水压是必不可少的，因为任何物理伤害可能会对鱼的生存和后期行为产生负面影响。仔细适应水族箱可降低发病死亡率，减轻运输冲击。饲养期间的观察包括（1）识别个人双鱼和（2）监测产卵，对鱼或鸡蛋没有负面影响，从而对研究种的生殖生态进行详细的调查。皮下注射可见植入弹性体（VIE）标签是用于随后鉴定个体鱼的精确方法，并且可以与大范围的鱼一起使用，对其生存和行为影响最小。如果研究物种是沉积粘蛋的底物，那么由聚氯乙烯（PVC）管道构建的人造巢场，加上可拆卸的防水片将有助于计数和监测鸡蛋，减少调查人员对巢穴的影响和鱼的保卫行为。虽然这种基本方法需要在研究文章中很少提及的技术，但它们是进行需要养殖野生鱼类的实验的基础。
Introduction
鱼类的形态学，生态学和行为学表现出明显的自适应进化1 。特别是与生殖相关的生态特征特别多样，其中大部分可以直接受个人健身的影响2 。为了深入了解导致不同鱼类独特特征演变的选择性压力，使用活鱼直接观察生殖和社会行为通常有助于证实理论假设。
然而，对鱼类的连续现场观察可能需要专门的难以维护的水下设备和设施。在这些情况下，野生捕捞的水族馆饲养的鱼的观察是有帮助3，4，5。此外，鱼类行为的有效观察，其他罕见或困难ULT在自然条件下，观察可以通过在水族箱6，7，8操纵实验成为可能。通过最大程度地减少人为压力和身体伤害，在良好条件下养鱼是准确生态调查的关键。
侏儒虾青y素总长 23-25毫米，分布在西太平洋，安静，庇护的海湾，深度9-26米9 。在这项工作中， T. marinae被用作一个模型，用于描述使用独立的水下呼吸器（SCUBA），鱼类运输和最终驯化鱼类进行直接观察的鱼类收集系列的基本技术的研究物种的生殖行为和生态学。
Protocol
收集和运送活鱼注意：本协议描述了如何从具有深度≥15米的气囊收集具有气囊的鱼。快速输送到表面会导致气囊膨胀，压力变化会严重危害或杀死鱼类。谨慎是必要的，因为在第一步中对鱼类造成的损害将对其生存和后期行为产生不利影响。 在与SCUBA潜水之前，请收集以下材料：适合捕获水下目标物种的手网;双重聚乙烯袋，足够大的鱼;橡胶带（ø80 mm x 6 mm）;用于在?…
Representative Results
按照上述方法，分别于2014年4月，2015年和2016年在日本鹿儿岛县Am美大岛境外收集了41，15和96个海棠的个体（ 表1 ）。在每种情况下，25（61％），14（93％）和91（95％）个人住在直到在水族馆里存放鸡蛋。福田等人报道3 ，在2014年观察期结束前只有一只鱼死亡，鱼类产卵否则在捕获后7天开始出现，表明个体在良好条件下饲?…
Discussion
许多鱼类的生殖生态常常通过实验饲养来揭示。特别是，变性6，8，14，择偶15,16和种内竞争7,17一直使用水族馆保持鱼类详细调查的频繁的话题。此外，在水族箱观察到一些结果已经在后面的字段8，18</sup…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
感谢S.Yokoyama协助收集鱼。我们也非常感谢瓦川村有关饲养方法的有用建议。本研究由日本科学促进会（KAKENHI）通过授予TS（24370006和16K07507）的资助
Materials
| Hand net | Nisso | AQ-17 | Select for the target species size. |
| Polyethylene bag | San-U Fish Farm | 8194 | |
| Rubber band | ESCO Co. LTD. | 78-0420-64 | ø 80 mm x 6 mm |
| Oxygen cylinder | N/A | N/A | Oxgen cylinder for diving equipment suits. |
| Elbagin | Japan Pet Design Co. Ltd. | 75950 | Pafurazine F (provided from same company) is equivalent drug to Elbagin. |
| Polystyrene foam box | N/A | N/A | |
| Pipette | AS ONE | 1-8625-04 | |
| Rope | Mizukami Kinzoku Co. LTD. | 95301601 | |
| Weight | N/A | N/A | Weight for diving equipment suits. |
| Water tank | N/A | N/A | |
| Air pump | KOTOBUKI | 4972814 062115 | |
| Air stone | KOTOBUKI | 4972814 232204 | |
| 2-Methylquinoline | Wako | 170-00376 | |
| Ethanol (99.5) | Wako | 057-00456 | |
| Visible implant elastomer tag kit | Northwest Marine Technology | N/A | http://www.nmt.us/products/vie/vie.shtml |
| Soft sponge | N/A | N/A | |
| PVC board | N/A | N/A | 0.3 mm thickness is easy to use. |
| Petri dish | N/A | N/A | A large one, such as ø 160 mm and 30 mm depth, is convenient for the injection of the VIE tag. |
| Transparent acrylic board | N/A | N/A | |
| UVA filtered light | N/A | N/A | |
| PVC pipe | N/A | N/A | ø 5 cm |
| Waterproof sheet | SOMAR Corp. | 3EKW03 | The film for the plain copier. |
| Sand | N/A | N/A | |
| Stereo microscope | N/A | N/A | |
| Camera | N/A | N/A | |
Riferimenti
- Helfman, G., Collette, B. B., Facey, D. E., Bowen, B. W. . The Diversity of Fishes: Biology, Evolution, and Ecology. , (2009).
- Davies, N. B., Krebs, J. R., West, S. A. . An Introduction to Behavioural Ecology. , (2012).
- Fukuda, K., Manabe, H., Sakurai, M., Dewa, S., Shinomiya, A., Sunobe, T. Monogamous mating system and sexuality in the gobiid fish, Trimma marinae (Actinopterygii: Gobiidae). J Ethol. 35 (1), 121-130 (2017).
- Manabe, H., Matsuoka, M., Goto, K., Dewa, S., Shinomiya, A., Sakurai, M., Sunobe, T. Bi-directional sex change in the gobiid fish Trimma sp.: does size-advantage exist?. Behaviour. 145 (1), 99-113 (2008).
- Sakurai, M., Nakakoji, S., Manabe, H., Dewa, S., Shinomiya, A., Sunobe, T. Bi-directional sex change and gonad structure in the gobiid fish Trimma yanagitai. Ichthyol Res. 56 (1), 82-86 (2009).
- Sunobe, T., Nakazono, A. Sex change in both directions by alteration of social dominance in Trimma okinawae (Pisces: Gobiidae). Ethology. 94 (4), 339-345 (1993).
- Wong, M. Y., Munday, P. L., Buston, P. M., Jones, G. P. Monogamy when there is potential for polygyny: tests of multiple hypotheses in a group-living fish. Behav Ecol. 19 (2), 353-361 (2008).
- Kuwamura, T., Suzuki, S., Tanaka, N., Ouchi, E., Karino, K., Nakashima, Y. Sex change of primary males in a diandric labrid Halichoeres trimaculatus: coexistence of protandry and protogyny within a species. J Fish Biol. 70 (6), 1898-1906 (2007).
- Winterbottom, R. Two new species of the Trimma tevegae species group from the Western Pacific (Percomorpha: Gobiidae). Aqua J Ichthyol Aquat Biol. 10, 29-38 (2005).
- Frederick, J. L. Evaluation of fluorescent elastomer injection as a method for marking small fish. Bull Mar Sci. 61 (2), 399-408 (1997).
- Olsen, E. M., Vøllestad, L. A. An evaluation of visible implant elastomer for marking age-0 brown trout. N Am J Fish Manag. 21 (4), 967-970 (2001).
- Leblanc, C. A., Noakes, D. L. Visible implant elastomer (VIE) tags for marking small rainbow trout. N Am J Fish Manag. 32 (4), 716-719 (2012).
- Kinkel, M. D., Eames, S. C., Philipson, L. H., Prince, V. E. Intraperitoneal injection into adult zebrafish. J Vis Exp. (42), e2126 (2010).
- Reavis, R. H., Grober, M. S. An integrative approach to sex change: social, behavioural and neurochemical changes in Lythrypnus dalli (Pisces). Acta Ethol. 2 (1), 51-60 (1999).
- Milinski, M., Bakker, T. C. Female sticklebacks use male coloration in mate choice and hence avoid parasitized males. Nature. 344, 330-333 (1990).
- Basolo, A. L. Phylogenetic evidence for the role of a pre-existing bias in sexual selection. Proc R Soc Lond B Biol Sci. 259 (1356), 307-311 (1995).
- Milinski, M. Optimal forging: the influence of intraspecific competition on diet selection. Behav Ecol Sociobiol. 11 (2), 109-115 (1982).
- Manabe, H., Ishimura, M., Shinomiya, A., Sunobe, T. Field evidence for bidirectional sex change in the polygynous gobiid fish Trimma okinawae. J Fish Biol. 70 (2), 600-609 (2007).
- Stammler, K. L., Corkum, L. D. Assessment of fish size on shelter choice and intraspecific interactions by round gobies Neogobius melanostomus. Environ Biol Fishes. 73 (2), 117-123 (2005).
- Matsumoto, Y., Tawa, A., Takegaki, T. Female mate choice in a paternal brooding blenny: the process and benefits of mating with males tending young eggs. Ethology. 117 (3), 227-235 (2011).
- McCormick, M. I. Behaviorally induced maternal stress in a fish influences progeny quality by a hormonal mechanism. Ecology. 79 (6), 1873-1883 (1998).
- Knaepkens, G., Bruyndoncx, L., Coeck, J., Eens, M. Spawning habitat enhancement in the European bullhead (Cottus gobio), an endangered freshwater fish in degraded lowland rivers. Biodivers Conserv. 13 (13), 2443-2452 (2004).
- Shiogaki, M., Dotsu, Y. The life history of the gobiid fish, Zonogobius boreus. Bulletin of the Faculty of Fisheries, Nagasaki University. 37, 1-8 (1974).
- Meunier, B., Yavno, S., Ahmed, S., Corkum, L. D. First documentation of spawning and nest guarding in the laboratory by the invasive fish, the round goby (Neogobius melanostomus). J Great Lakes Res. 35 (4), 608-612 (2009).
Citazione di questo articolo
Fukuda, K., Sunobe, T. Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria. J. Vis. Exp. (125), e55964, doi:10.3791/55964 (2017).