Name: Construction of an Affordable and Easy-to-Build Zebrafish Facility (Video) | JoVE Journal
Uploaded: 2014-11-22T14:00:00.000+00:00
Duration: 12 min 19 s
Description: Learn how to build an affordable zebrafish facility using PVC components, pumps, and filters for neuroscience and developmental biology research. This method enables student involvement and requires minimal maintenance.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

The authors would like to thank the Office of Research of Presbyterian College School of Pharmacy for financial support of this project through the Small Pharmacy Awards for Research &#38; Collaboration (SPARC) Award. The authors thank Mr. John Smink, Manager of the Aquatic Animal Research Laboratory, Clemson University, Clemson, SC for his technical assistance manuscript revision.

NOTE: These protocols follow the guidelines of IACUC of Presbyterian College.
1. Acquire Materials Listed in Table 1.
2. Assemble the 5 Shelf Chrome According to the Manufacturer&#8217;s Instructions. This will be used to House the Facility.
3. Construct the water reservoir and filtration system (Figures 1A and 2).
<ol>
	<li>Using Figure 1A as a guide, layout the 18 gallon bucket, pump, filters and UV lamp to determine final installation location and size.</li>
	<li>Once parts are laid out, measure and cut the &#190;&#8221; PVC pipe to the length needed for assembly. Vary the lengths of all pipes according to specific space needs and limitations.</li>
	<li>Assemble the pipes and fittings as laid out previously using the PVC primer and cement, see below (3.3.2 and 3.3.3).
	<ol>
		<li>Sand the cut edges of the pipe with sandpaper until they are smooth to the touch and there is no debris.</li>
		<li>Prime the outside of the pipe and the inside of the fitting with the purchased PVC primer with the included swab from the PVC primer.</li>
		<li>Cement the pipe and fitting together by applying the PVC cement to the outside of the pipe using the included swab. Repeat for all steps that need PVC cementing.</li>
		<li>Place the 18 gallon bucket to one side of the shelf construction.</li>
		<li>Install a bulkhead connector in the side of the reservoir tank.</li>
		<li>Attach a &#190;&#8221; PVC pipe to the bulkhead.</li>
		<li>Attach the other end of the &#190;&#8221; pipe to the pump inlet using a &#190;&#8221; threaded make fitting.</li>
		<li>Connect the filters to the pump using two 90 degree elbows and &#190;&#8221; PVC pipe.
		<ol>
			<li>Use one paper filter followed by one activated carbon filters in order to filter out debris and reduce chlorine and chemical levels in the water.</li>
		</ol>
		</li>
		<li>Connect the UV lamp to the filters according to the manufacturer&#8217;s instructions.</li>
	</ol>
	</li>
	<li>Attach the pump and filter system to the top side of the first shelf of the rack using wire ties. Vary the order of the assembly and attachment according to the size and layout of the shelf/rack chosen.</li>
	<li>Add 1 bioball per gallon of water to the water reservoir once the pump and filter system are assembled and attached to the shelf.</li>
</ol>
4. Construct the main supply line (Figures 1B and 2).
<ol>
	<li>Measure and cut a piece of &#190;&#8221; PVC pipe to connect the UV lamp to the main supply</li>
	<li>Use a &#190;&#8221; PVC tee to start the main supply line by connecting the tee to the &#190;&#8221; PVC exhaust coming from the UV filter. .
	<ol>
		<li>Add an on-off valve to the bottom of the PVC tee to serve as a drain for maintenance flushing of the system.</li>
		<li>Start the main supply line from the top of the &#190;&#8221; PVC tee by measuring and cutting &#190;&#8221; PVC pipe to desired length to provide feed pipes for each of the shelves on the rack.</li>
		<li>Install &#190;&#8221; PVC tees for the horizontal lines that will enter each shelf of the rack.</li>
		<li>Optionally (and suggested), add a &#190;&#8221; on-off valve for each shelf supply line to allow for isolation of each shelf and a modular design of the system.</li>
	</ol>
	</li>
	<li>Use a &#190;&#8221; PVC pipe as a pressure reducing line on the top shelf. If this line runs on the back of the shelf, use this shelf for storage. End the pressure reducing line in the main exhaust line (Figure 1B) with a &#190;&#8221; on-off valve.</li>
	<li>Once the above steps have been completed, prime and cement the PVC pipe together as previously done when constructing the reservoir and filtration system.</li>
</ol>
5. Construct the main exhaust line (Figures 1C and 2).
<ol>
	<li>Run the vertical length of the main exhaust line to the other side of the rack from the main supply line. Use a 2&#8221; PVC pipe for this vertical line to receive the main supply line at the top and drain lines from the fish tanks from each shelf.</li>
	<li>Cut the pieces of 2&#8221; PVC pipe for the vertical main exhaust line and use 2&#8221; PVC tees to receive the fish tank drain lines from each shelf. End the main exhaust line in the stock tank to expel the recycled water over a sponge filter.</li>
	<li>Cut a one inch opening the entire length of the 2&#8221; PVC pipe pieces and attach to vertical main exhaust line with PVC tees for the horizontal drain lines from the fish tanks.</li>
	<li>Cap the 2&#8221; horizontal drain lines with a 2&#8221; PVC cap.</li>
	<li>Prime and cement the pieces using the necessary fittings (connectors, nipples or elbows).</li>
</ol>
6. Construct the fish tanks (Figure 3).
<ol>
	<li>With the drill, cut a 1 1/8&#8221; hole at the back of the fish tank 1&#8221; below the upper edge with a hole saw.</li>
	<li>Slide a threaded &#190;&#8221; PVC male adapter (with a #18 O ring attached) through the hole (from the inside of the tank to the outside).</li>
	<li>Attach and tighten a 90 degree 3/4&#39; threaded PVC elbow to the male adapter to create the fish tank exhaust pipe. Add a piece of fine mesh before screwing the holding nut to prevent zebrafish going through the exhaust system.</li>
	<li>Place the fish tank with its exhaust pipe ending into the open space of the draining line behind the tanks</li>
</ol>
7. Starting the water system
<ol>
	<li>In several containers, such as the one used for the reservoir, measure three times the amount of water contained within the system. Let this water stand at room temperature for conditioning for 48 hr.</li>
	<li>Add water to the system and let it circulate for a minimum of 3 days.
	<ol>
		<li>Change the water in the system vigorously throughout the initial 3 days of circulation in order to rid the system of contaminants that reside within the pipes.</li>
	</ol>
	</li>
	<li>Add starter zebrafish and allow the nitrogen cycle to get established. Normalize pH to a consistent 7. Keep seven zebrafish per 1.8 L, and feed them twice daily during week days and once daily during weekends as suggested by Linbo4 and McNabb6.</li>
	<li>Monitor the nitrate (&#60;75 mg/L), nitrite (&#60;10 mg/L), and ammonium (0 mg/L) daily during the first week after adding fish, and twice weekly once desired levels have been maintained with regular water changes.</li>
</ol>

<ol><li>Detrich, H., Westerfield, M., Zon, L. <a target="_blank" href="http://scholar.google.com/scholar?hl=en&safe=off&q=author%3aDetrich%2C+H+%22The+Zebrafish%3A+Cellular+and+Developmental+Biology%2C+Part+A.%22">The Zebrafish: Cellular and Developmental Biology, Part A.</a>. , 3rd ed, Academic Press. New York, New York. (2010).</li>
<li>Grim, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Hemogen+%28hemgn%29%2C+a+transcription+factor+that+regulates+erythropoiesis+and+skeletogenesis+in+zebrafish%5BTitle%5D)+AND+%22%22%5BJournal%5D)">Hemogen (hemgn), a transcription factor that regulates erythropoiesis and skeletogenesis in zebrafish.</a> Strategic Conference of Zebrafish Investigators Poster Presentation, 2013, Asilimar, CA, , (2013).</li>
<li>Kim, S., Carlson, R., Zafreen, L., Rajpurohit, S. K., Jagadeeswaran, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Modular%2C+easy-to-assemble%2C+low-cost+facility%5BTitle%5D)+AND+%22Zebrafish%22%5BJournal%5D)">Modular, easy-to-assemble, low-cost facility.</a> Zebrafish. 6 (3), 269-274 (2009).</li>
<li>Linbo, T. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Zebrafish+%28Danio+rerio%29+husbandry+and+colony+maintenance+at+the+Northwest+Fisheries+Science+Center%5BTitle%5D)+AND+%22U.S.+Dept.+Commer.%2C+NOAA+Tech.+Memo%22%5BJournal%5D)">Zebrafish (Danio rerio) husbandry and colony maintenance at the Northwest Fisheries Science Center.</a> U.S. Dept. Commer., NOAA Tech. Memo. 62, (2009).</li>
<li>Avdesh, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Regular+care+and+maintenance+of+a+zebrafish+%28Danio+rerio%29+laboratory%3A+an+introduction%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Regular care and maintenance of a zebrafish (Danio rerio) laboratory: an introduction.</a> J Vis Exp. 69, 4196(2012).</li>
<li>McNabb, A., Scott, K., Ochsenstein, E., Seufert, K., Carl, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Don%27t+be+afraid+to+set+up+your+fish+facility%5BTitle%5D)+AND+%22Zebrafish%22%5BJournal%5D)">Don't be afraid to set up your fish facility.</a> Zebrafish. 9 (3), 120-125 (2012).</li>
<li>Garcia, R. L., Sanders, G. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Efficacy+of+cleaning+and+disinfection+procedures+in+a+zebrafish+rerio%29+facility%5BTitle%5D)+AND+%22J+Am+Assoc+Lab+Anim+Sci%22%5BJournal%5D)">Efficacy of cleaning and disinfection procedures in a zebrafish rerio) facility.</a> J Am Assoc Lab Anim Sci. 50 (6), 895-900 (2011).</li>
<li>Rivas-Boyero, A. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Pharmacological+characterization+of+a+nociceptin+receptor+from+zebrafish+%28Danio+rerio%29%5BTitle%5D)+AND+%22J+Mol+Endocrinol%22%5BJournal%5D)">Pharmacological characterization of a nociceptin receptor from zebrafish (Danio rerio).</a> J Mol Endocrinol. 46 (2), 111-123 (2011).</li>
<li>Sanchez-Simon, F. M., Zhang, X. X., Loh, H. H., Law, P. Y., Rodriguez, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Morphine+regulates+dopaminergic+neuron+differentiation+via+miR-133b%5BTitle%5D)+AND+%22Mol+Pharmacol%22%5BJournal%5D)">Morphine regulates dopaminergic neuron differentiation via miR-133b.</a> Mol Pharmacol. 78 (5), 935-942 (2010).</li>
<li>Velasco, E. M., Law, P. Y., Rodríguez, R. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Mu+opioid+from+the+zebrafish+exhibits+functional+characteristics+as+those+of+mammalian+mu+opioid+receptor%5BTitle%5D)+AND+%22Zebrafish%22%5BJournal%5D)">Mu opioid from the zebrafish exhibits functional characteristics as those of mammalian mu opioid receptor.</a> Zebrafish. 6 (3), 259-268 (2009).</li>
</ol>

The authors declare no conflicts of interest for this paper.

The main purpose of this article is to demonstrate that the construction of a functional zebrafish facility is not complicated and can be built by novice fish researchers with minimal construction knowledge. Furthermore, this article shows that the materials for this functional zebrafish facility can be acquired readily and affordably from local pet supply stores or hardware stores. Once all the materials and tools have been acquired the construction of the system and the fish tanks takes about one working day. The 72 hr...

In vivo models are essential in conducting comprehensive biomedical research since they allow in vitro research to be translated at the system and organismal level. For example, research with small mammals, namely rodents, has been proven necessary in the development of drugs and in the improvement of available drugs. However, the complexity (financially and logistically) might make it virtually impossible to build and maintain rodent facilities for experimentation proposes at small research academic institutions. Nonetheless, in vivo research can be performed using other animal species that require less elaborate logistics. Zebrafish (Danio rerio) have been successfully used as an in vivo model to study genetics, development, pathophysiology, and pharmacology1,2,10,11,12. The facilities required to house zebrafish for experimentation are relatively simple and significantly less expensive when compared to rodent facilities. Kim et al.3 have recently described in detail an 80 tank zebrafish housing system that can be built for approximately $1,500, which includes both parts and labor costs. This stands in stark contrast to the nearly $8,500 that would be required to purchase a ready-made, 80 tank system from any number of commercial suppliers. However, the description given by Kim et al. may be complicated and difficult to follow for novices of fish research. We believe that a zebrafish facility can be constructed in simplified manner for an investment of $1,500.
The establishment of an in vivo facility at small academic institutions provides their faculties an additional tool with which to produce more powerful and thorough research. In vitro data supported by in vivo results generally produce more comprehensive biomedical research than data obtained using only in vitro approaches. Therefore, the implementation of an in vivo research facility will significantly enhance the institution success in its overall research endeavors (student exposure to research, publications, communications, pursuit of external funding, etc.).
The construction of a Zebrafish facility like the one described in this study is ideal for research academic institutions with limited facilities (or institutions without in vivo research facilities) during constricting financial times.

<table><tbody><tr><td>Item</td><td>Brand</td><td>Quantity</td><td></td></tr><tr><td>74-in H x 48-in W x 18-in D 5-shelf Steel Freestanding Shelving Unit</td><td>Style Selections</td><td>1</td><td></td></tr><tr><td>18-Gallon General Bucket/Tote</td><td>Centrex Plastics: LLC&amp;nbsp;Rugged Tote</td><td>1</td><td></td></tr><tr><td>6-Outlet Power Strip With 15-Ft. Cord</td><td>Master Electrician</td><td>1</td><td></td></tr><tr><td>9-Watt UV Water Garden Clarifier FiIter and UV lamp</td><td>Smartpond&amp;nbsp;</td><td>1</td><td></td></tr><tr><td>Whole House Water Replacement Filter</td><td>Whirlpool</td><td>6</td><td></td></tr><tr><td>Opaque Whole-House Pre-Filtration Housing</td><td>Whirlpool</td><td>2</td><td></td></tr><tr><td>Thread Seal Tape</td><td>William H Harvey</td><td>1</td><td></td></tr><tr><td>8-fl oz LO-VOC PVC Cement</td><td>Oatey&amp;nbsp;</td><td>1</td><td></td></tr><tr><td>8-fl oz LO-VOC Primer</td><td>Oatey&amp;nbsp;</td><td>1</td><td></td></tr><tr><td>1/4-in x 100-ft Vinyl Drip Irrigation Distribution Tubing</td><td>Mister Landscaper&amp;nbsp;</td><td>1</td><td></td></tr><tr><td>1/4-in Barbed Drip Irrigation On/Off Valve</td><td>Mister Landscaper</td><td>33</td><td></td></tr><tr><td>9-Port NPT Irrigation Manifold with Filter</td><td>&amp;nbsp;Mister Landscaper</td><td>6</td><td></td></tr><tr><td>Drop-In Filter with House&amp;nbsp;</td><td>Whirlpool</td><td>6</td><td></td></tr><tr><td>2-in Dia 90-Degree PVC Elbow</td><td>LASCO</td><td>3</td><td></td></tr><tr><td>2-in Dia 90-Degree Tee</td><td>LASCO</td><td>3</td><td></td></tr><tr><td>3/4-in Dia 90-Degree Slip Elbow</td><td>LASCO</td><td>15</td><td></td></tr><tr><td>3/4-in Dia PVC Adapter</td><td>LASCO</td><td>7</td><td></td></tr><tr><td>3/4-in Dia PVC Tee</td><td>LASCO</td><td>12</td><td></td></tr><tr><td>2-in Dia PVC Cap</td><td>LASCO</td><td>3</td><td></td></tr><tr><td>1-in Dia x 3/4-in Dia PVC Adapter</td><td>LASCO</td><td>2</td><td></td></tr><tr><td>3/4-in PVC Socket In-Line Ball Valve</td><td>American Valve</td><td>5</td><td></td></tr><tr><td>1/2-in Dia x 3/4-in Dia PVC Adapter</td><td>Lasco</td><td>8</td><td></td></tr><tr><td>3/4-in Dia PVC Cap</td><td>Lasco</td><td>1</td><td></td></tr><tr><td>2-In x 5-Ft Pvc Pipe</td><td>Charlotte Pipe</td><td>4</td><td></td></tr><tr><td>&amp;frac34;-In x 10-Ft Pvc pipe</td><td>Charlotte Pipe</td><td>3</td><td></td></tr><tr><td>11/16x1-7/80 PVC ring</td><td>SXT</td><td>10</td><td></td></tr><tr><td>&amp;frac12;&amp;rdquo; PVC male adapter</td><td>SXT</td><td>1</td><td></td></tr><tr><td>&amp;frac34;&amp;rdquo; PVC coupling&amp;nbsp;</td><td>SXT</td><td>1</td><td></td></tr><tr><td>&amp;frac12;&amp;rdquo; PVC female adapter&amp;nbsp;</td><td>SXT</td><td>1</td><td></td></tr><tr><td>link</td><td></td><td></td><td></td></tr><tr><td>1.8 L Dual Beta Keeper</td><td>Top Fin</td><td>27</td><td></td></tr><tr><td>&amp;frac34;&amp;rdquo; PVC male adapter (w/ #18 O-ring)</td><td>SXT</td><td>28</td><td></td></tr><tr><td>90-degree &amp;frac34;&amp;rdquo; threaded PVC elbow</td><td>SXT</td><td>27</td><td></td></tr><tr><td>Fine Mesh</td><td>N/A</td><td>1 yard</td><td></td></tr><tr><td>Bulkhead Connector</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Hacksaw</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Drill</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Hole saw</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Adjustable Wrench</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Pliers&amp;nbsp;</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Tape Measure</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Safety Glasses</td><td>N/A</td><td>1</td><td></td></tr><tr><td>Work Gloves</td><td>N/A</td><td>1</td><td></td></tr></tbody></table>

construction of an affordable and easy-to-build zebrafish facility

Construction of an Affordable and Easy-to-Build Zebrafish Facility

In vivo biomedical research is pivotal to translate in vitro findings into clinical advances. Small academic institutions with limited resources find it virtually impossible to build and maintain typical rodent facilities for research. Zebrafish research has been demonstrated to be a valuable alternative for in vivo research in pharmacology, physiology, development and genetic studies. This article demonstrates that a functional zebrafish facility can be built in an easy and affordable manner. We demonstrate that such a facility could be built in about one working day with minimal tools and expertise. The cost of the 27 1.8 L fish tank zebrafish facility constructed in this study was approximately $1,500. We estimate that the maintenance of an initial working 150 fish colony for 3 months is $1,000. This project involved students, who were introduced to aquaculturing of zebrafish for research proposes.

Construction of the Pump and Flters System 
Once the rack is assembled and set up the first step is to build the pump and filter system. The filters are connected to the system pump and are located above the bottom level of the first shelf, which allows for easy filter maintenance and replacement. Similarly, the UV lamp is connected with PVC pipe to the filter and held in place with plastic wire ties to allow for easy replacement of the UV lamp as needed (Figure 4A). Immediately following the...

A Zebrafish facility suitable for breeding and in vivo research is built in an easy and affordable manner. Maintenance is minimal. This facility is ideal for student involvement and independent research.

In vivo biomedical research is pivotal to translate in vitro findings into clinical advances. Small academic institutions with limited resources find it ...

construction-of-an-affordable-and-easy-to-build-zebrafish-facility

Research

JoVE Journal

Biology

This protocol outlines regular maintenance and care to maintain optimal conditions for zebrafish husbandry. The video illustrates the protocol for system maintenance, regular housing, feeding, breeding, and raising of zebrafish larvae.

Regular Care and Maintenance of a Zebrafish (Danio rerio) Laboratory: An Introduction

The zebrafish embryo is an excellent model for developmental biology research. During embryogenesis, zebrafish develop with a yolk mass, which presents three-dimensional challenges for sample observation and analysis. This protocol describes how to create two-dimensional flat mount preparations of whole mount in situ (WISH) stained zebrafish embryo specimens.

Flat Mount Preparation for Observation and Analysis of Zebrafish Embryo Specimens Stained by Whole Mount In situ Hybridization

The zebrafish is an excellent experimental organism to study vertebrate developmental processes and model human disease. Here, we describe a protocol on how to perform a manual high-throughput chemical screen in zebrafish embryos with a whole-mount in situ hybridization (WISH) read-out.

A Manual Small Molecule Screen Approaching High-throughput Using Zebrafish Embryos

Microinjection of zebrafish embryos and larvae is a crucial but challenging technique used in many zebrafish models. Here, we present a range of microscale tools to aid in the stabilization and orientation of zebrafish for both microinjection and imaging.

Microstructured Devices for Optimized Microinjection and Imaging of Zebrafish Larvae

Here, we present a protocol to design and fabricate a zebrafish embryo arraying template, followed by a detailed procedure on the use of such template for high throughput zebrafish embryo arraying into a 96-well plate.

A 3-dimensional (3D)-printed Template for High Throughput Zebrafish Embryo Arraying

This work describes a protocol for the modular Tol2 transgenesis system, a gateway-based cloning method to create and inject transgenic constructs into zebrafish embryos.

The Zebrafish Tol2 System: A Modular and Flexible Gateway-Based Transgenesis Approach

The exercise apparatus designed for less fortunate fish facilitates the implementation of diverse exercise protocols with varying intensities by manipulating water flow speed, achievable through rheotaxis.

A Swimming-Induced Zebrafish Exercise Apparatus for Versatile Training Approaches

Here, we present a novel, easy-to-construct, and low-cost protocol for recirculating zebrafish housing.

A Custom-Built Recirculating Zebrafish Housing System

Keeping the microscope optics clean is important for high-quality imaging. Dust, fingerprints, excess immersion oil, or mounting medium on or in a microscope causes reduction in contrast and resolution. DIC is especially sensitive to contamination and scratches on the lens surfaces. This protocol details the procedure for keeping the microscope clean.

Proper Care and Cleaning of the Microscope

A short protocol for protein staining with Coomassie Brilliant Blue (CBB) G-250 in polyacrylamide gels is described without using organic solvents or acetic acid as in the classical staining procedures with CBB.

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

Learn how to build an affordable zebrafish facility using PVC components, pumps, and filters for neuroscience and developmental biology research. This method enables student involvement and requires minimal maintenance.

Construction of an Affordable and Easy-to-Build Zebrafish Facility

Explore More Videos