Name: basic methods for the study of reproductive ecology of fish in aquaria
Uploaded: 2017-07-20T13:00:00
Description: Watch this Scientific Journal Video about Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria at JoVE.com

We thank S. Yokoyama for assistance with collecting fish. We are also grateful to W. Kawamura for helpful advice about rearing methods. This study was supported by the Japan Society for the Promotion of Science (KAKENHI) through grants (No. 24370006 and 16K07507) awarded to T.S.

1. Collecting and Transporting Live Fish
NOTE: This protocol describes how to collect fish that possess a gas bladder, from a depth of &#8805;15 m to the surface. Rapid conveyance to the surface will induce expansion of the gas bladder by a change of pressure, which can seriously harm or kill the fish. Caution is warranted, as damage caused to the fish during this first step will negatively effect their survival and later behavior.
<ol>
	<li>Prior to diving with SCUBA, gather the following m...

<ol>
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The reproductive ecology of numerous fishes has often been revealed through experimental rearing. Especially, sex change6,8,14, mate choice15,16 and intraspecific competition7,17 have been frequent topics of detailed investigations using aquarium-kept fish. Furthermore, some results observed in aquaria have been ...

Spectacular adaptive evolution is evident in the morphology, ecology and behavior of fishes1. Especially, ecological features relating to reproduction are especially diverse, and most of these can be directly influenced by individual fitness2. To gain insight into selective pressures that have led to the evolution of unique features in different fish species, direct observation of reproductive and social behaviors using live fish is often beneficial to substantiate theoretical hypotheses.
However, continuous field observations of fish may require specialized underwater equipment and facilities that are difficult to maintain. In these cases, observations of wild-caught aquarium-reared fish can be helpful3,4,5. In addition, efficient observations of fish behaviors that are otherwise rare or difficult to observe under natural conditions can become possible by manipulating experiments in aquaria6,7,8. Rearing fish under good conditions by minimizing artificial stress and physical damage is critical for accurate ecological investigations.
The pygmy goby Trimma marinae reaches 23-25 mm total length and is distributed in the western Pacific Ocean, where it is found in quiet, sheltered bays, at depths of 9-26 m9. In this work, T. marinae is used as a model to describe a series of basic techniques for the collection of fish using the self-contained underwater breathing apparatus (SCUBA), fish transport, and eventual acclimation of the fish to aquaria for direct observation of the study species&#39; reproductive behavior and ecology.

<table border="0" cellpadding="0" cellspacing="0" width="1037">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:192px;">Hand net</td>
			<td style="width:193px;">Nisso</td>
			<td style="width:108px;">AQ-17</td>
			<td style="width:544px;">Select for the target species size.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polyethylene bag</td>
			<td>San-U Fish Farm</td>
			<td>8194</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rubber band</td>
			<td>ESCO Co. LTD.</td>
			<td>78-0420-64</td>
			<td>&#248; 80 mm x 6 mm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oxygen cylinder</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Oxgen cylinder for diving equipment suits.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Elbagin</td>
			<td>Japan Pet Design Co. Ltd.</td>
			<td>75950</td>
			<td>Pafurazine F (provided from same company) is equivalent drug to Elbagin.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Polystyrene foam box</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pipette</td>
			<td>AS ONE</td>
			<td>1-8625-04</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Rope</td>
			<td>Mizukami Kinzoku Co. LTD.</td>
			<td>95301601</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Weight</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>Weight for diving equipment suits.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Water tank</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Air pump</td>
			<td>KOTOBUKI</td>
			<td colspan="2">4972814 062115</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Air stone</td>
			<td>KOTOBUKI</td>
			<td colspan="2">4972814 232204</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">2-Methylquinoline&#160;</td>
			<td>Wako</td>
			<td>170-00376</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Ethanol (99.5)</td>
			<td>Wako</td>
			<td>057-00456</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Visible implant elastomer tag kit</td>
			<td>Northwest Marine Technology</td>
			<td>N/A</td>
			<td>http://www.nmt.us/products/vie/vie.shtml</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Soft sponge</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVC board</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>0.3 mm thickness is easy to use.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Petri dish</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>A large one, such as&#160; &#248; 160 mm and 30 mm depth, is convenient for the injection of the VIE tag.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Transparent acrylic board</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UVA filtered light&#160;</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PVC pipe</td>
			<td>N/A</td>
			<td>N/A</td>
			<td>&#248; 5 cm</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Waterproof sheet</td>
			<td>SOMAR Corp.</td>
			<td>3EKW03</td>
			<td>The film for the plain copier.</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Sand</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stereo microscope</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Camera</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
	</tbody>
</table>

basic methods for the study of reproductive ecology of fish in aquaria

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a series of basic techniques are described to enable observations of the reproductive behavior of a wild-caught gobiid fish, as a model, kept in an aquarium. The method focuses on three steps: collection, transport, and observations of reproductive ecology of a substrate spawner. Essential aspects of live fish collection and transport are (1) preventing injury to the fish, and (2) careful acclimation to the aquarium. Preventing harm through injuries such as scratches or a sudden change of water pressure is imperative when collecting live fish, as any physical damage is likely to negatively affect the survival and later behavior of the fish. Careful acclimation to aquaria decreases the incidence death and mitigates the shock of transport. Observations during captive rearing include (1) the identification of individual fish and (2) monitoring spawned eggs without negative effects to the fish or eggs, thereby enabling detailed investigation of the study species' reproductive ecology. The subcutaneous injection of a visible implant elastomer (VIE) tag is a precise method for the subsequent identification of individual fish, and it can be used with a wide size range of fish, with minimal influence on their survival and behavior. If the study species is a substrate spawner that deposits adhesive eggs, an artificial nest site constructed from polyvinyl chloride (PVC) pipe with the addition of a removable waterproof sheet will facilitate counting and monitoring the eggs, lessening the investigator's influence on the nest-holding and egg-guarding behavior of the fish. Although this basic method entails techniques that are seldom mentioned in detail in research articles, they are fundamental for undertaking experiments that require the captive rearing of a wild fish.

Following the above methods, 41, 15 and 96 individuals of T. marinae were collected in April 2014, 2015 and 2016, respectively, offshore of Amami Oshima, Kagoshima Prefecture, Japan (Table 1). In each case, 25 (61%), 14 (93%) and 91 (95%) individuals lived until depositing eggs in an aquarium. As reported in Fukuda et al.3, only one fish died before the end of the observation period in 2014, and fish spawning otherwise appeared to...

Watch this Scientific Journal Video about Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria at JoVE.com

Basic Methods for the Study of Reproductive Ecology of Fish in Aquaria

A series of basic methods to enable the study of the reproductive ecology of fish kept in aquaria are described. These are useful protocols for collecting fish using SCUBA, transporting live fish, and observing the reproductive behavior of wild-caught fish kept in aquaria.

Captive-rearing observations are valuable for revealing aspects of fish behavior and ecology when continuous field investigations are impossible. Here, a ...

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