Name: effectiveness of the air stripping in two salmonid fish, rainbow trout (oncorhynchus mykiss) and brown trout (salmo trutta morpha fario)
Uploaded: 2018-09-16T13:00:00.000+00:00
Duration: 3 min 50 s
Description: Watch this Scientific Journal Video about Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout Oncorhynchus Mykiss and Brown Trout Salmo Trutta Morpha fario at JoVE.com

The presented study was supported by Project "Pneumatic method of fish stripping - possible application, influence on the gametes' quality and quantity and the welfare of fish" (acronym: PNEUFISH) financed under "Operational Program Development of the Fisheries Sector and Coastal Areas 2007-2013" (OR-61724-OR1400001/10), funds appropriated to Institute of Animal Reproduction and Food Research Polish Academy of Sciences, Olsztyn, Poland, and support by COST Office (Food and Agriculture COST Action FA1205: AQUAGAMETE). We would like to thank, Stanley Ifeanyi Ugwu for great support in English editing. We want to acknowledge also the animation producers "Studio Filmowe Ruchome Obrazki".

Procedures were carried out in accordance with the Local Committee on the Ethics of Animal Experiments in Olsztyn, Poland.
1.Equipment Preparation
<ol>
	<li>Use a syringe and needle for injection of the gas into the fish body cavity.</li>
	<li>Set the gas pressure at 0.5 bar, and maintain this pressure to be below 1 bar throughout.</li>
	<li>Control the air flow. For the salmonid fish, 1.5 L/min is the most efficient speed of gas exhaustion. Maintain the optimum speed because higher speed might not obtain all the eggs while a slower speed might unnecessarily prolong the procedure.</li>
	<li>Connect the syringe with a screwed needle (0.8-1.2 mm diameter) to avoid needle detachment caused by the gas pressure.</li>
</ol>
2. Air Spawning
<ol>
	<li>Anesthetize the fish with 0.2% Propiscin. Place the fish into a water bath with the 0.2% Propiscin and wait until the fish becomes unresponsive.</li>
	<li>Carefully dry out before spawning by wiping with a dry cotton cloth.</li>
	<li>Place the fish on the slant bed about 35-40&#176;, on her side tail side down. With a gentle push of fish abdomen, check the egg flow from the genital tract. If the eggs are not present, the fish should not be spawned.</li>
	<li>Place a container covered with mesh below the fish papilla to collect the eggs.</li>
	<li>Quickly insert the needle below the abdominal fins.</li>
	<li>Freely remove the eggs by the gas until only singular eggs are removed from the fish body.</li>
	<li>Gently massage the abdomen to remove excess gas from the fish body.</li>
	<li>After spawning, gently transfer the fish to a tank with water flow. Fish released to tank after air spawning might initially swim on the surface of the water. After several minutes the fish releases the remaining gas while swimming and starts swimming normally. 
	NOTE: The obtained eggs are very clean and free from blood or feces contamination.</li>
</ol>

<ol>
	<li>Nelson, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Fishes of the World. , (2006).</li><li>Garc&#237;a-Mar&#237;n, J. L., Sanz, N., Pla, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proportions+of+native+and+introduced+brown+trout+in+adjacent+fished+and+unfished+Spanish+rivers.">Proportions of native and introduced brown trout in adjacent fished and unfished Spanish rivers.</a> Conservation Biology. 12, 313-319 (1998).</li><li>Dietrich, G. J., Wojtczak, M., Slowinska, M., Dobosz, S., Kuzminski, H., Ciereszko, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Broken+eggs+decrease+pH+of+rainbow+trout+(Oncorhynchus+mykiss)+ovarian+fluid.">Broken eggs decrease pH of rainbow trout (Oncorhynchus mykiss) ovarian fluid.</a> Aquaculture. 273, 748-751 (2007).</li><li>Zurbuch, P. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+structure+for+easy+fish+recovery+during+drainage+of+an+impoundment.">A structure for easy fish recovery during drainage of an impoundment.</a> Progressive FishCulturist. 27, 237-238 (1965).</li><li>Cejko, B. I., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+different+stripping+methods+of+female+and+activation+medium+on+fertilization+success+in+northern+pike+(Esox+lucius).">Effects of different stripping methods of female and activation medium on fertilization success in northern pike (Esox lucius).</a> Czech Journal of Animal Sciences. 10, 481-486 (2016).</li><li>Morisawa, S., Morisawa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induction+of+potential+for+sperm+motility+by+bicarbonate+and+pH+in+rainbow+trout+and+chum+salmon.">Induction of potential for sperm motility by bicarbonate and pH in rainbow trout and chum salmon.</a> Journal of Experimental Biology. 136, 13-22 (1988).</li><li>Billard, R., Cosson, J., Perchec, G., Linhart, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biology+of+sperm+and+artificial+reproduction+in+carp.">Biology of sperm and artificial reproduction in carp.</a> Aquaculture. 129, 95-112 (1995).</li><li>Iuchi, I., Ha, C. R., Sugiyama, H., Nomura, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+chorion+hardening+of+eggs+of+rainbow+trout,+Oncorhynchus+mykiss.">Analysis of chorion hardening of eggs of rainbow trout, Oncorhynchus mykiss.</a> Development Growth and Differentiation. 38, 299-306 (1996).</li><li>Wojtczak, M. A., Kowalski, R. K., Dobosz, S., Goryczko, K., Ku&#378;minski, H., Glogowski, J., Ciereszko, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+water+turbidity+for+evaluation+of+rainbow+trout+(Oncorhynchus+mykiss)+egg+quality.">Assessment of water turbidity for evaluation of rainbow trout (Oncorhynchus mykiss) egg quality.</a> Aquaculture. 242, 617-624 (2004).</li><li>Tabrizi, E. N., Khara, H., Nezami, S. A., Lorestani, R., Shamspour, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Broken+Eggs+Influence+on+Fertilization+Capacity+and+Viability+of+Eggs,+Turbidity+and+pH+of+Ovarian+Fluid+and+Fertilization+Water+in+the+Endangered+Caspian+Brown+Trout,+Salmo+Trutta+Caspius.">Broken Eggs Influence on Fertilization Capacity and Viability of Eggs, Turbidity and pH of Ovarian Fluid and Fertilization Water in the Endangered Caspian Brown Trout, Salmo Trutta Caspius.</a> International Journal of Biology. 3 (1), 161-166 (2011).</li><li>Carl, G. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beware+of+the+broken+egg!+A+possible+cause+of+heavy+losses+of+salmon+eggs.">Beware of the broken egg! A possible cause of heavy losses of salmon eggs.</a> Progresive Fish-Culturist. 53, 30-31 (1941).</li></ol>

The authors have nothing to disclose.

The pneumatic method of egg collection from fish, although more time-consuming than the traditional method (hand stripping), can secure a high quality of matured oocytes (this work). This is related to the low risk of mechanical disruption of the eggs while applying this procedure. High pH of the ovary fluid, as well as an increase in the fertilization rate, attest to the usefulness of the pneumatic method. Overall, the air stripping method of egg collection in fish reproduction could be a promising option in terms of st...

The rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta morpha fario) belong to the Salmonidae family of Salmoniformes1. The production of these two species in global aquaculture has gained rapid growth, owing to their commercial and recreational importance. In Poland, production of salmonid fish stands at around 20 000 tonnes, with rainbow trout being the dominant species. On the other hand, the brown trout represents a major source of freshwater fish resources in Europe because of its commercial value in aquaculture and importance for angling. In many aquatic ecosystems, the existence of brown trout populations is threatened. As a result, artificial reproduction has been applied to restock local populations of this species2.
Rainbow trout and brown trout attain sexual maturity usually at the age of three, with males maturing a year earlier than females1. Under artificial conditions, eggs and sperm of rainbow trout and brown trout are usually collected by gentle abdominal massage. In practice, on a small scale, a hand-stripping method of collecting eggs is cost-effective. However, on a large scale, the hand-stripping method may be labor-intensive and exhausting. As a result, this technique can result in broken eggs due to the weariness of the hatchery workers. Breakage of eggs often leads to a decrease in the pH of ovarian fluid and the release of egg yolk, both of which negatively affect the fertilization of eggs3. Furthermore, this traditional method of egg collection (repeatedly pressing of the abdomen) may also result in broodstock mortality caused by skin infection associated with the deterioration of the protective barrier of mucus. 
Pneumatic method of egg collection was first used in Australia in 19574. The method is based on the injection of gases (air, nitrogen or oxygen) into the body cavity to expel eggs using gas pressure. This fast and easy technique has been successfully applied in salmonid fish without any negative side effects on the fish4. Recently, this method was used in wild species (Northern pike, Esox lucius) to collect mature eggs in artificial conditions5. It was shown that air stripped eggs had higher fertilization and hatching rates in comparison with hand stripped eggs, regardless of the solution used for fertilization (Woynarovich solution, Billard buffer or hatchery water)5.
The principal aim of this study was to test the pneumatic method (air stripping) of collecting eggs in rainbow trout and brown trout in comparison with traditional hand stripping method. Efficiencies of both methods (volume of obtained eggs and time of collection, quality of gametes (ovarian fluid pH), as well as hatching rate and post-spawning mortality) were compared.

<table><tbody><tr><td>Glycine pure P.A.</td><td>AVANTOR</td><td>527560117</td><td>sperm activating buffer</td></tr><tr><td>Trizma base</td><td>Sigma Aldrich</td><td>T1503</td><td>sperm activating buffer</td></tr><tr><td>Sodium chloride Bioreagent</td><td>Sigma Aldrich</td><td>S5886</td><td>sperm activating buffer</td></tr><tr><td>Calcium chloride anhydrous</td><td>Sigma Aldrich</td><td>C4901</td><td>sperm activating buffer</td></tr><tr><td>Propiscin (0.2% etomidate solution)</td><td>IRS Olsztyn, Poland</td><td>not indicated</td><td></td></tr><tr><td>compressor</td><td>Thomas&amp;nbsp; Sheboigan WI USA&amp;nbsp;</td><td>DT/SR 070800001882</td><td></td></tr><tr><td>reduction valve</td><td>Camozzi&amp;nbsp;</td><td>cRJUS U7J</td><td></td></tr><tr><td>air hose</td><td>ZEC T.P.U.</td><td>&amp;nbsp;SH.98&amp;nbsp;</td><td></td></tr><tr><td>syrgine</td><td>EFD</td><td>7012118</td><td></td></tr><tr><td>Air spawning stage</td><td>Biopasz, Poland</td><td>PNEU001</td><td></td></tr><tr><td>Orion Ross Ultra electrode</td><td>Thermo Scientific, Waltham, MA, USA</td><td>8102BNUWP</td><td></td></tr></tbody></table>

effectiveness of the air stripping in two salmonid fish, rainbow trout (oncorhynchus mykiss) and brown trout (salmo trutta morpha fario)

Egg collection is one of the most crucial procedures during fish reproduction in salmonid hatcheries. Classic methods involve the use of hand massage on fish abdomens to expel the eggs. An alternative method uses the pressure of gas injected into the body cavity, which causes the subsequent release of the eggs. This method is believed to have less negative effects on both the welfare and egg quality of the broodstocks. Herein, we compare the results of air and hand stripping methods with respect to one-year survival and egg quantity and quality in two salmonid fish, rainbow trout (Oncorhynchus mykiss) and brown trout (Salmo trutta morpha fario). Our results indicate that air stripping yielded a better quality of eggs and higher one-year survival rate in rainbow trout. In addition, air stripping resulted in lower mortality rate than the group subjected to hand stripping (25% vs. 35%). The pH and hatching rate of the hand stripped group was lower than those of the air stripped group. In the case of brown trout, the quality of eggs obtained by both hand and air-stripping methods was similar; however, the one-year losses in fish were higher in air stripped group (15% compared to 0% in hand stripped fish). Although the advantages of air stripping method over hand stripping in terms of egg quality might not be observed in all salmonid species, the air-stripping procedure might be a promising option to be adopted in hatcheries as it ensures a high level of reproducibility and efficiency.

Mature males and females of rainbow trout (age 2+, 1700 &#177; 328 g) and brown trout (age 3+, 1900 &#177; 435 g) were obtained from the Salmonid Fish Hatchery (Rutki-&#379;ukowo, Inland Fishery Institute in Olsztyn, Poland). The males and females were placed in separate tanks with a volume of about 5 m3. The water temperature was 12&#177;1 &#176;C. Before manipulation, fish were anesthetized with 0.2% Propiscin. The fish were divided into two groups. Part of them ...

Watch this Scientific Journal Video about Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout Oncorhynchus Mykiss and Brown Trout Salmo Trutta Morpha fario at JoVE.com

Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout Oncorhynchus Mykiss and Brown Trout Salmo Trutta Morpha fario

The principal aim of this study is to standardize and test the pneumatic method (air stripping) of collecting eggs in rainbow trout and brown trout. This method allows effective and simple collection of the eggs without the necessity of fish abdomen massage.

Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout (Oncorhynchus Mykiss) and Brown Trout (Salmo Trutta Morpha fario)

Egg collection is one of the most crucial procedures during fish reproduction in salmonid hatcheries. Classic methods involve the use of hand massage on ...

effectiveness-air-stripping-two-salmonid-fish-rainbow-trout

Research

JoVE Journal

Developmental Biology

7.8K Views.  Polish Academy of Science. The principal aim of this study is to standardize and test the pneumatic method (air stripping) of collecting eggs in rainbow trout and brown trout. This method allows effective and simple collection of the eggs without the necessity of fish abdomen massage.

Video: Effectiveness of the Air Stripping in Two Salmonid Fish, Rainbow Trout Oncorhynchus Mykiss and Brown Trout Salmo Trutta Morpha fario

Laboratory Estimation of Net Trophic Transfer Efficiencies of PCB Congeners to Lake Trout (Salvelinus namaycush) from Its Prey

A technique for laboratory estimation of net trophic transfer efficiency (&#947;) of polychlorinated biphenyl (PCB) congeners to piscivorous fish from their prey is described herein. During a 135-day laboratory experiment, we fed bloater (Coregonus hoyi) that had been caught in Lake Michigan to lake trout (Salvelinus namaycush) kept in eight laboratory tanks. Bloater is a natural prey for lake trout. In four of the tanks, a relatively high flow rate was used to ensure relatively high activity by the lake trout, whereas a low flow rate was used in the other four tanks, allowing for low lake trout activity. On a tank-by-tank basis, the amount of food eaten by the lake trout on each day of the experiment was recorded. Each lake trout was weighed at the start and end of the experiment. Four to nine lake trout from each of the eight tanks were sacrificed at the start of the experiment, and all 10 lake trout remaining in each of the tanks were euthanized at the end of the experiment. We determined concentrations of 75 PCB congeners in the lake trout at the start of the experiment, in the lake trout at the end of the experiment, and in bloaters fed to the lake trout during the experiment. Based on these measurements, &#947; was calculated for each of 75 PCB congeners in each of the eight tanks. Mean &#947; was calculated for each of the 75 PCB congeners for both active and inactive lake trout. Because the experiment was replicated in eight tanks, the standard error about mean &#947; could be estimated. Results from this type of experiment are useful in risk assessment models to predict future risk to humans and wildlife eating contaminated fish under various scenarios of environmental contamination.

Laboratory Estimation of Net Trophic Transfer Efficiencies of PCB Congeners to Lake Trout Salvelinus namaycush from Its Prey

A technique for laboratory estimation of net trophic transfer efficiency of polychlorinated biphenyl (PCB) congeners to piscivorous fish from their prey is presented. To maximize applicability of the laboratory results to the field, the piscivorous fish should be fed prey fish that are typically eaten in the field.

A technique for laboratory estimation of net trophic transfer efficiency (&#947;) of polychlorinated biphenyl (PCB) congeners to piscivorous fish from ...

Environment

Use of a Battery of Chemical and Ecotoxicological Methods for the Assessment of the Efficacy of Wastewater Treatment Processes to Remove Estrogenic Potency

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in a watch list of environmental pollutants under the European Water Framework Directive. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. Much of the estrogenic potency of wastewater treatment plant (WWTP) effluents can be attributed to the discharge of steroid estrogens including estradiol (E2), EE2 and E1 due to incomplete removal of these substances at the treatment plant. An evaluation of the efficacy of wastewater treatment processes requires the quantitative determination of individual substances most often undertaken using chemical analysis methods. Most frequently used methods include Gas Chromatography-Mass Spectrometry (GCMS/MS) or Liquid Chromatography-Mass Spectrometry (LCMS/MS) using multiple reaction monitoring (MRM). Although very useful for regulatory purposes, targeted chemical analysis can only provide data on the compounds (and specific metabolites) monitored. Ecotoxicology methods additionally ensure that any by-products produced or unknown estrogenic compounds present are also assessed via measurement of their biological activity. A number of in vitro bioassays including the Yeast Estrogen Screen (YES) are available to measure the estrogenic activity of wastewater samples. Chemical analysis in conjunction with in vivo and in vitro bioassays provides a useful toolbox for assessment of the efficacy and suitability of wastewater treatment processes with respect to estrogenic endocrine disrupting compounds. This paper utilizes a battery of chemical and ecotoxicology tests to assess conventional, advanced and emerging wastewater treatment processes in laboratory and field studies.

Endocrine Disrupting Compounds (EDC) pose a substantial risk to the aquatic environment. Municipal wastewater treatment plants are major contributors to the estrogenic potency of surface waters. The methodology provided in this paper allows for an assessment of the efficacy and suitability of wastewater treatment processes with respect to EDC removal.

Endocrine Disrupting Compounds pose a substantial risk to the aquatic environment. Ethinylestradiol (EE2) and estrone (E1) have recently been included in ...

A Flow-through Exposure System for Evaluating Suspended Sediments Effects on Aquatic Life

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory by using pumps and automating delivery of sediment and water to simulate suspension of sediment. FLEES data are used to develop exposure-response curves between the effects on aquatic organisms and suspended sediment concentrations at the desired exposure duration. The effects data are used to evaluate management practices used to reduce the interactions between aquatic organisms and anthropogenic causes of suspended sediments. The FLEES is capable of generating total suspended solids (TSS) concentrations as low as 30 to as high as 800 mg/L, making this system an ideal choice for evaluating the effects of TSS resulting from many activities including simulating low ambient levels of TSS to evaluating sources of suspended sediments from dredging operations, vessel traffic, freshets, and storms.

A robust and flexible flow-through exposure system designed to maintain sediment in suspension is presented. The system is used to investigate the effects of suspended sediment on various aquatic species and life stages in the laboratory.

This paper describes the Fish Larvae and Egg Exposure System (FLEES). The flow-through exposure system is used to investigate the effects of suspended ...

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity testing has been demonstrated. Overall, the sensitivity of the species to toxic compounds is in the range with, or higher than, that of other model species.
This work describes protocols for acute, chronic, and multigenerational bioassays of single and combined stressor effects on N. furzeri. Due to its short maturation time and life-cycle, this vertebrate model allows the study of endpoints such as maturation time and fecundity within four months. Transgenerational full life-cycle exposure trials can be performed in as little as 8 months. Since this species produces eggs that are drought-resistant and remain viable for years, the on-site culture of the species is not needed but individuals can be recruited when required. The protocols are designed to measure life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

Protocol for Acute and Chronic Ecotoxicity Testing of the Turquoise Killifish Nothobranchius furzeri

In this work, we describe an acute, chronic and multigenerational bioassay to study the effects of single and combined stressors on the Turquoise killifish Nothobranchius furzeri. This protocol is designed to study life-history traits (mortality, growth, fecundity, weight) and critical thermal maximum.

The killifish Nothobranchius furzeri is an emerging model organism in the field of ecotoxicology and its applicability in acute and chronic ecotoxicity ...

A Standardized Protocol for Preference Testing to Assess Fish Welfare

Animal welfare assessment techniques try to take into consideration the specific needs and wants of the animal in question. Providing enrichment (the addition of physical objects or conspecifics in the housing environment) is often a way to give captive animals the opportunity to choose who or what they interact with and how they spend their time. A fundamental component of the aquatic environment that is often overlooked in captivity, however, is the ability for the animal to choose to engage in physical exercise. For many animals, including fish, exercise is an important aspect of their life history, and is known to have many health benefits, including positive changes in the brain and behavior. Here we present a method for assessing habitat preferences in captive animals. The protocol could easily be adapted to look at a variety of environmental factors (e.g., gravel versus sand as a substrate, plastic plants versus live plants, low flow versus high flow of water) in different aquatic species, or for use with terrestrial species. Statistical assessment of preference is carried out using Jacob&#39;s preference index, which ranks the habitats from -1 (avoidance) to +1 (most preferred). With this information, it can be determined what the animal wants from a welfare perspective, including their preferred location.

A fundamental aspect of assessing the welfare of animals in captivity is to ask whether the animals have what they want. Here, we present a protocol to determine housing preference in the zebrafish (Danio rerio) with respect to the presence/absence of environmental enrichment and access to flowing of water.

Animal welfare assessment techniques try to take into consideration the specific needs and wants of the animal in question. Providing enrichment (the ...

Behavior

A Novel Method of Drug Administration to Multiple Zebrafish (Danio rerio) and the Quantification of Withdrawal

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely netted and transported between home aquaria and dosing tanks. In order to enhance the ease of compound administration, a novel method for transferring fish between tanks for drug administration was developed. Inserts that are designed for spawning were used to transfer groups of fish into the drug solution, allowing accurate dosing of all fish in the group. This increases the precision and efficiency of dosing, which becomes very important in long schedules of repeated drug administration. We implemented this procedure for use in a study examining the behavior of zebrafish in the light/dark test after administering ethanol with differing 21 day schedules. In fish exposed to daily-moderate amounts of alcohol there was a significant difference in location preference after 2 days of withdrawal when compared to the control group. However, a significant difference in location preference in a group exposed to weekly-binge administration was not observed. 
This protocol can be generalized for use with all types of compounds that are water-soluble and may be used in any situation when the behavior of fish during or after long schedules of drug administration is being examined. The light/dark test is also a valuable method of assessing withdrawal-induced changes in anxiety.

A Novel Method of Drug Administration to Multiple Zebrafish Danio rerio and the Quantification of Withdrawal

A novel method for reducing variability when exposing fish to drugs is explained. Fish exposed to various patterns of ethanol exposure were found to have altered anxiety levels during withdrawal in a light/dark scototaxic assay.

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely ...

Neuroscience

Modeling Fetal Alcohol Spectrum Disorders in Zebrafish to Characterize the Impact of an Adverse Embryonic Environment on Adult Social Behavior

Fetal alcohol spectrum disorders (FASD) describe all alcohol-induced birth defects. Birth defects such as growth deficiencies, craniofacial, behavioral, and cognitive abnormalities are associated with FASD. Social difficulties are common behavioral abnormalities associated with FASD and often result in serious health issues. Animal models are critical to understanding the mechanisms responsible for ethanol-induced social defects. Zebrafish are social vertebrates that produce externally fertilized transparent eggs; these characteristics provide researchers with a precise yet simple procedure for creating the FASD phenotype and an innate behavior that can be leveraged to model the social deficits associated with FASD. Thus, zebrafish are ideal for characterizing the social deficits of FASD. The goal of the current protocol is to provide the user with a simple behavioral assay that can be used to characterize the consequences of a negative environment early during development and the effects it can have on social behavior in adulthood. The protocol can be used to characterize the effect mutations or teratogens have on adult social behavior. The protocol outlined here demonstrates how to characterize the social behavior of individual fish during a 20-min social assay. Furthermore, the data obtained using the current protocol provides evidence that the protocol can be used to characterize the effects of embryonic ethanol-induced social defects in adult zebrafish.

The goal of the current protocol is to outline the steps necessary to establish and use a social preference assay for adult zebrafish and demonstrate that it can be used to characterize ethanol-induced social defects.

Fetal alcohol spectrum disorders (FASD) describe all alcohol-induced birth defects. Birth defects such as growth deficiencies, craniofacial, behavioral, ...

Quantifying Fish Swimming Behavior in Response to Acute Exposure of Aqueous Copper Using Computer Assisted Video and Digital Image Analysis

Behavioral responses of aquatic organisms to environmental contaminants can be precursors of other effects such as survival, growth, or reproduction. However, these responses may be subtle, and measurement can be challenging. Using juvenile white sturgeon (Acipenser transmontanus) with copper exposures, this paper illustrates techniques used for quantifying behavioral responses using computer assisted video and digital image analysis. In previous studies severe impairments in swimming behavior were observed among early life stage white sturgeon during acute and chronic exposures to copper. Sturgeon behavior was rapidly impaired and to the extent that survival in the field would be jeopardized, as fish would be swept downstream, or readily captured by predators. The objectives of this investigation were to illustrate protocols to quantify swimming activity during a series of acute copper exposures to determine time to effect during early lifestage development, and to understand the significance of these responses relative to survival of these vulnerable early lifestage fish. With mortality being on a time continuum, determining when copper first affects swimming ability helps us to understand the implications for population level effects. The techniques used are readily adaptable to experimental designs with other organisms and stressors.

Measuring the impacts of environmental contaminants on fish behavior is often subjective and challenging particularly when dealing with sublethal endpoints. We describe methods including video technology to quantify swimming behavior of early life stage white sturgeon (Acipenser transmontanus) during and after 96 hr acute exposures to various concentrations of copper.

Behavioral responses of aquatic organisms to environmental contaminants can be precursors of other effects such as survival, growth, or reproduction.  ...

Swimming Performance Assessment in Fishes

Swimming performance tests of fish have been integral to studies of muscle energetics, swimming mechanics, gas exchange, cardiac physiology, disease, pollution, hypoxia and temperature. This paper describes a flexible protocol to assess fish swimming performance using equipment in which water velocity can be controlled. The protocol involves one to several stepped increases in flow speed that are intended to cause fish to fatigue. Step speeds and their duration can be set to capture swimming abilities of different physiological and ecological relevance. Most frequently step size is set to determine critical swimming velocity (Ucrit), which is intended to capture maximum sustained swimming ability. Traditionally this test has consisted of approximately ten steps each of 20 min duration. However, steps of shorter duration (e.g. 1 min) are increasingly being utilized to capture acceleration ability or burst swimming performance. Regardless of step size, swimming tests can be repeated over time to gauge individual variation and recovery ability. Endpoints related to swimming such as measures of metabolic rate, fin use, ventilation rate, and of behavior, such as the distance between schooling fish, are often included before, during and after swimming tests. Given the diversity of fish species, the number of unexplored research questions, and the importance of many species to global ecology and economic health, studies of fish swimming performance will remain popular and invaluable for the foreseeable future.

The lives of the majority of fish are predicated on swimming. This protocol describes techniques for capturing a range of swimming modes available to individual and schooling fish, and includes metrics associated with swimming physiology and behaviour.

Swimming performance tests of fish have been integral to studies of muscle energetics, swimming mechanics, gas exchange, cardiac physiology, disease, ...

Biology

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

Health monitoring systems are developed and used in zebrafish research facilities because pathogens of Danio rerio such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa have the potential to impair animal welfare and research. The fish are typically analyzed post mortem to detect microbes. The use of sentinels is a suggested way to improve the sensitivity of the surveillance and to reduce the number of animals to sample. The setting of a pre-filtration sentinel tank out of a recirculating system is described. The technique is developed to prevent water pollution and to represent the fish population by a careful selection of age, gender, and strains. In order to use the minimum number of animals, techniques to screen the environment are also detailed. Polymerase Chain Reaction (PCR) on surface sump swabs is used to significantly improve the detection of some prevalent and pathogenic mycobacterial species such as Mycobacterium fortuitum, Mycobacterium haemophilum, and Mycobacterium chelonae. Another environmental method consists of processing the sludge at the bottom of a holding tank or sump to look for P. tomentosa eggs. This is a cheap and fast technique that can be applied in quarantine where a breeding device is submerged into the holding tank of imported animals. Finally, PCR is applied to the sludge sample and A. hydrophila is detected at the sump&#39;s bottom and surface. Generally, these environmental screening techniques applied to these specific pathogens have led to an increased sensitivity compared to the testing of pre-filtration sentinels.

Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems

This protocol describes the use of sump swabs and sludge analysis of zebrafish systems, which leads to increased detection compared to the sole use of sentinels to detect pathogens such as Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa. A system to monitor P. tomentosa eggs in quarantine is also proposed.

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Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems