Skip to main content
Log in

Stress test of a biological early warning system with zebrafish (Danio rerio)

  • Published:
Ecotoxicology Aims and scope Submit manuscript

Abstract

The aim of this work was to develop a novel methodology to stress test the diagnostic capability of a video tracking system with zebrafish (Danio rerio), against two pre-established disturbances. Eight different treatments were tested varying the presence or absence of a toxicant (NaOCl) and two disturbances: the passing of a shadow (mimicking a predator) and entrapment of the fish. The concentration tested corresponded to a sublethal (1 % 24 h-LC50) and short term exposure (2 h). A total of 56 organisms were tested resulting in 112 diagnoses (before and after the contamination). A statistical model of diagnosis was developed using Self-organizing Map (SOM) and Correspondence Analysis (CA). Sensitivity, specificity, accuracy, false positive and false negative values were calculated to evaluate the diagnostic performance. The disturbances did not negatively affect the capability of the model. In the presence of at least one of these variables, the diagnostic performance was similar or even superior to the baseline results without disturbances. Furthermore, the system produced a large number of correct diagnoses, at an ecologically relevant concentration of exposure, in a non-invasive way.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  • Bae MJ, Park YS (2014) Biological early warning system based on the responses of aquatic organisms to disturbances: a review. Sci Total Environ 466-467:635–649

    Article  CAS  Google Scholar 

  • Bailey B, Amre DK (2005) A toxicologist’s guide to studying diagnostic tests. Clin Toxicol (Phila) 43:171–179

    Article  Google Scholar 

  • Blaser R, Gerlai R (2006) Behavioral phenotyping in zebrafish: comparison of three behavioral quantification methods. Behav Res Methods 38:456–469

    Article  Google Scholar 

  • Chen T-H, Wang Y-H, Wu Y-H (2011) Developmental exposures to ethanol or dimethylsulfoxide at low concentrations alter locomotor activity in larval zebrafish: Implications for behavioral toxicity bioassays. Aquat Toxicol 102:162–166

    Article  CAS  Google Scholar 

  • Chon TS, Chung N, Kwak IS, Kim JS, Koh SC, Lee SK, Leem JB, Cha EY (2005) Movement behaviour of Medaka (Oryzias latipes) in response to sublethal treatments of diazinon and cholinesterase activity in semi-natural conditions. Environ Monit Assess 101:1–21

    CAS  Google Scholar 

  • Chon T-S, Park YS, Moon KH, Cha EY (1996) Patternizing communities by using an artificial neural network. Ecol Model 90:69–78

    Article  Google Scholar 

  • Cunha SR, Goncalves R, Silva SR, Correia AD (2008) An automated marine biomonitoring system for assessing water quality in real-time. Ecotoxicology 17:558–564

    Article  Google Scholar 

  • Decreto-Lei 113/2013 do Ministério da Agricultura, do Mar, do Ambiente e do Ordenamento do Território de 7 de Agosto de 2013 relativa à proteção dos animais utilizados para fins científicos (2013)

  • Delcourt J, Becco C, Ylieff MY, Caps H, Vandewalle N, Poncin P (2006) Comparing the EthoVision 2.3 system and a new computerized multitracking prototype system to measure the swimming behavior in fry fish. Behav Res Methods 38:704–710

    Article  Google Scholar 

  • Directive 2010/63/EU of the European Parlament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes (2010) L 276/33

  • Dolado R, Gimeno E, Beltran FS, Quera V, Pertusa JF (2015) A method for resolving occlusions when multitracking individuals in a shoal. Behav Res Methods 47:1032–1043

    Article  Google Scholar 

  • Gerlai R, Csanyi V (1990) Genotype-environment interaction and the correlation structure of behavioral elements in paradise fish (Macropodus opercularis). Physiol Behav 47:343–356

    Article  CAS  Google Scholar 

  • Gerlai R, Fernandes Y, Pereira T (2009) Zebrafish (Danio rerio) responds to the animated image of a predator: Towards the development of an automated aversive task. Behav Brain Res 201:318–324

    Article  Google Scholar 

  • Ghisleni G, Capiotti KM, Da Silva RS, Oses JP, Piato AL, Soares V, Bogo MR, Bonan CD (2012) The role of CRH in behavioral responses to acute restraint stress in zebrafish. Prog Neuro-psychopharmacol Biol Psychiatry 36:176–182

    Article  CAS  Google Scholar 

  • Gomez-Laplaza LM, Gerlai R (2010) Latent learning in zebrafish (Danio rerio). Behav Brain Res 208:509–515

    Article  Google Scholar 

  • Grillitsch B, Vogl C, Wytek R (1999) Qualification of spontaneous undirected locomotor behavior of fish for sublethal toxicity testing. Part II. Variability of measurement parameters under toxicant-induced stress. Environ Toxicol Chem 18:2743–2750

    Article  CAS  Google Scholar 

  • Halkin A, Reichman J, Schwaber M, Paltiel O, Brezis M (1998) Likelihood ratios: getting diagnostic testing into perspective. QJM Int J Med 91:247–258

    Article  CAS  Google Scholar 

  • Kane AS, Salierno JD, Gipson GT, Molteno TCA, Hunter C (2004) A video-based movement analysis system to quantify behavioral stress responses of fish. Water Res 38:3993–4001

    Article  CAS  Google Scholar 

  • Knottnerus JA, van Weel C, Muris JW (2002) Evaluation of diagnostic procedures. Br Med J (Clin Res Ed) 324:477–480

    Article  Google Scholar 

  • Kohonen T (2001) Self-organizing maps, 3rd edition. Springer, Berlin, p 347–371

    Google Scholar 

  • Kühl AM, da Rocha MSC, Lucia C, Gaeta Espíndola EL, Lansac-Tôha FA (2010) Rural and urban streams: anthropogenic influences and impacts on water and sediment quality. Int Rev Hydrobiol 95:260–272

    Article  Google Scholar 

  • Little EE, Finger SE (1990) Swimming behavior as an indicator of sublethal toxicity in fish. Environ Toxicol Chem 9:13–19

    Article  CAS  Google Scholar 

  • Luca RM, Gerlai R (2012) In search of optimal fear inducing stimuli: Differential behavioral responses to computer animated images in zebrafish. Behav Brain Res 226:66–76

    Article  Google Scholar 

  • Magalhães P, Armando da Cunha R, Albuquerque dos Santos JA, Buss DF, Baptista DF (2007) Behavioral response of zebrafish Danio rerio Hamilton 1822 to sublethal stress by sodium hypochlorite: ecotoxicological assay using an image analysis biomonitoring system. Ecotoxicology 16:417–422

    Article  Google Scholar 

  • Nimkerdphol K, Nakagawa M (2008) Effect of sodium hypochlorite on zebrafish swimming behavior estimated by fractal dimension analysis. J Biosci Bioeng 105:486–492

    Article  CAS  Google Scholar 

  • Oliva Teles L, Fernandes M, Amorim J, Vasconcelos V (2015) Video-tracking of zebrafish (Danio rerio) as a biological early warning system using two distinct artificial neural networks: probabilistic neural network (PNN) and self-organizing map (SOM). Aquat Toxicol 165:241–248

    Article  CAS  Google Scholar 

  • Park Y-S, Céréghino R, Compin A, Lek S (2003) Applications of artificial neural networks for patterning and predicting aquatic insect species richness in running waters. Ecol Model 160:265–280

    Article  Google Scholar 

  • Pather S, Gerlai R (2009) Shuttle box learning in zebrafish (Danio rerio). Behav Brain Res 196:323–327

    Article  Google Scholar 

  • Piato AL, Rosemberg DB, Capiotti KM, Siebel AM, Herrmann AP, Ghisleni G, Vianna MR, Bogo MR, Lara DR, Bonan CD (2011) Acute restraint stress in zebrafish: behavioral parameters and purinergic signaling. Neurochem Res 36:1876–1886

    Article  CAS  Google Scholar 

  • Richards FM, Alderton WK, Kimber GM, Liu Z, Strang I, Redfern WS, Valentin JP, Winter MJ, Hutchinson TH (2008) Validation of the use of zebrafish larvae in visual safety assessment. J Pharmacol Toxicol Methods 58:50–58

    Article  CAS  Google Scholar 

  • van der Schalie WH, Shedd TR, Knechtges PL, Widder MW (2001) Using higher organisms in biological early warning systems for real-time toxicity detection. Biosens Bioelectron 16:457–465

    Article  Google Scholar 

  • Scott GR, Sloman KA (2004) The effects of environmental pollutants on complex fish behaviour: integrating behavioural and physiological indicators of toxicity. Aquat Toxicol 68:369–392

    Article  CAS  Google Scholar 

  • Scott GR, Sloman KA, Rouleau C, Wood CM (2003) Cadmium disrupts behavioural and physiological responses to alarm substance in juvenile rainbow trout (Oncorhynchus mykiss). J Exp Biol 206:1779–1790

    Article  CAS  Google Scholar 

  • Sirtes G, Zehnder M, Schaetzle M, Waltimo T (2005) The effects of temperature on sodium hypochlorite short-term stability, pulp dissolution capacity, and antimicrobial efficacy. J Endod 31:669–671

    Article  Google Scholar 

  • StatSoft (2012) STATISTICA (data analysis software system) (version 11. www.statsoft.com.)

  • van Stralen KJ, Stel VS, Reitsma JB, Dekker FW, Zoccali C, Jager KJ (2009) Diagnostic methods I: sensitivity, specificity, and other measures of accuracy. Kidney Int 75:1257–1263

    Article  Google Scholar 

  • Tahedl H, Hader DP (2001) Automated biomonitoring using real time movement analysis of Euglena gracilis. Ecotoxicol Environ Saf 48:161–169

    Article  CAS  Google Scholar 

  • Tahedl H, Häder D-P (1999) Fast examination of water quality using the automatic biotest ECOTOX based on the movement behavior of a freshwater flagellate. Water Res 33:426–432

    Article  CAS  Google Scholar 

  • Williams LR, Wong K, Stewart A, Suciu C, Gaikwad S, Wu N, DiLeo J, Grossman L, Cachat J, Hart P, Kalueff AV (2012) Behavioral and physiological effects of RDX on adult zebrafish. Comp Biochem Physiol Part C Toxicol Pharmacol 155:33–38

    Article  CAS  Google Scholar 

  • Winter MJ, Redfern WS, Hayfield AJ, Owen SF, Valentin J-P, Hutchinson TH (2008) Validation of a larval zebrafish locomotor assay for assessing the seizure liability of early-stage development drugs. J Pharmacol Toxicol Methods 57:176–187

    Article  CAS  Google Scholar 

  • Wong K, Elegante M, Bartels B, Elkhayat S, Tien D, Roy S, Goodspeed J, Suciu C, Tan J, Grimes C, Chung A, Rosenberg M, Gaikwad S, Denmark A, Jackson A, Kadri F, Chung KM, Stewart A, Gilder T, Beeson E, Zapolsky I, Wu N, Cachat J, Kalueff AV (2010) Analyzing habituation responses to novelty in zebrafish (Danio rerio). Behav Brain Res 208:450–457

    Article  CAS  Google Scholar 

  • Zhang K, Wang JZ, Liang B, Shen RL, Zeng EY (2012) Assessment of aquatic wastewater pollution in a highly industrialized zone with sediment linear alkylbenzenes. Environ Toxicol Chem 31:724–730

    Article  Google Scholar 

Download references

Acknowledgments

This research was partially funded by UID/Multi/04423/2013 project from Fundação para a Ciência e Tecnologia.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to João Amorim.

Ethics declarations

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Amorim, J., Fernandes, M., Vasconcelos, V. et al. Stress test of a biological early warning system with zebrafish (Danio rerio). Ecotoxicology 26, 13–21 (2017). https://doi.org/10.1007/s10646-016-1736-5

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10646-016-1736-5

Keywords

Navigation