Use of Locomotor Activity of Rainbow Trout Juveniles in Identifying Sublethal Concentrations of Landfill Leachate
Landfill waste is a common problem as it has an economic and environmental impact even if it is closed. Landfill waste contains a high density of various persistent compounds such as heavy metals, organic and inorganic materials. As persistent compounds are slowly-degradable or even non-degradable in the environment, they often produce sublethal or even lethal effects on aquatic organisms. The aims of the present study were to estimate sublethal effects of the Kairiai landfill (WGS: 55°55‘46.74“, 23°23‘28.4“) leachate on the locomotor activity of rainbow trout Oncorhynchus mykiss juveniles using the original system package developed in our laboratory for automated monitoring, recording and analysis of aquatic organisms’ activity, and to determine patterns of fish behavioral response to sublethal effects of leachate. Four different concentrations of leachate were chosen: 0.125; 0.25; 0.5 and 1.0 mL/L (0.0025; 0.005; 0.01 and 0.002 as part of 96-hour LC50, respectively). Locomotor activity was measured after 5, 10 and 30 minutes of exposure during 1-minute test-periods of each fish (7 fish per treatment). The threshold-effect-concentration amounted to 0.18 mL/L (0.0036 parts of 96-hour LC50). This concentration was found to be even 2.8-fold lower than the concentration generally assumed to be “safe” for fish. At higher concentrations, the landfill leachate solution elicited behavioral response of test fish to sublethal levels of pollutants. The ability of the rainbow trout to detect and avoid contaminants occurred after 5 minutes of exposure. The intensity of locomotor activity reached a peak within 10 minutes, evidently decreasing after 30 minutes. This could be explained by the physiological and biochemical adaptation of fish to altered environmental conditions. It has been established that the locomotor activity of juvenile trout depends on leachate concentration and exposure duration. Modeling of these parameters showed that the activity of juveniles increased at higher leachate concentrations, but slightly decreased with the increasing exposure duration. Experiment results confirm that the behavior of rainbow trout juveniles is a sensitive and rapid biomarker that can be used in combination with the system for fish behavior monitoring, registration and analysis to determine sublethal concentrations of pollutants in ambient water. Further research should be focused on software improvement aimed to include more parameters of aquatic organisms’ behavior and to investigate the most rapid and appropriate behavioral responses in different species. In practice, this study could be the basis for the development and creation of biological early-warning systems (BEWS).
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 U. K. Singh, M. Kumar, R. Chauhan, P. K. Jha A. Ramanathan, V. Subramanian, “Assessment of the impact of landfill on groundwater quality: A case study of the Pirana site in western India,” Environmental Monitoring and Assessment, vol. 141, no. 1, pp. 309-321, June 2008.
 S. Kanmani, R. Gandhimathi, “Assessment of heavy metal contamination in soil due to leachate migration from an open dumping site,” Applied Water Science, vol. 3, no. 1, pp. 193-205, March 2013.
 S. Kanmani, R. Gandhimathi, “Investigation of physicochemical characteristics and heavy metal distribution profile in groundwater system around the open dump site,” Applied Water Science, vol. 3, no. 2, pp. 387-399, June 2013.
 M. Y. J. Alkassasbeh, L. Y. Heng, S. Surif, “Toxicity testing and the effect of landfill leachate in Malaysia on behavior of common carp (Cyprinus Carpio L., 1758; Pisces, Cyprinidae),” American Journal of Environmental Sciences, vol. 5, no. 3, pp. 209-217, 2009.
 C. R Klauck, M. A Rodrigues, L. B. da Silva, “Toxicological evaluation of landfill leachate using plant (Allium cepa) and fish (leporinus obtusidens) bioassays,” Waste Management and Research, vol. 31 no. 11, pp. 1148–1153, November, 2013.
 G. Svecevičius, N. Kazlauskienė, A. Slučkaitė, T. Makaras, “Toxicological assessment of the effects of closed landfill on neighbouring hydroecosystem,” Fressenius Environmental Bulletin, vol. 23 no.11a, pp. 2926–2932, 2014.
 J. Kalka, “Landfill leachate toxicity removal in combined treatment with municipal wastewater,“ Scientific World Journal, 2012; 2012: 202897, doi:10.1100/2012/202897.
 J. Derco, A. Ž. Gotvajn, J. Zagorc-Končan, B. Almasiova, A. Kassai, “Pretreatment of landfill leachate by chemical oxidation processes,” Chemical Papers, vol. 64, no. 2, pp. 237-245, April 2010.
 Q. B. Gu, S. S. Liu, X. N. Zhuang, X. J. Li, F. S. Li, “Preparation and performance of inorganic coagulant for landfill Leachate Pretreatment,” Bulletin of Environmental Contamination and Toxicology, vol. 76, pp. 98-104, 2006.
 B. Jezierska, M. Witeska, “Metal toxicity to fish”, Wydawnictwo akademii Podlaskiej, University of Podlasie, Siedlce, pp. 12, 2001.
 R. Vinodhini, M. Narayanan “Bioaccumulation of heavy metals in organs of fresh water fish Cyprinus carpio (Common carp),” International Journal of Environmental Science and Technology, vol 5, no. 2, pp. 179-182, March 2008.
 G. J. Atchison, M. G. Henry, M. B. Sandheinrich, ”Effects of metals on fish behavior: a review,“ Environmental Biology of Fishes, vol. 18, pp. 11-25, 1987.
 J. Y. M. Alkassasbeh, L. Y. Heng, S. Surif, ”Toxicity testing and the effect of landfill leachate in Malaysia on behavior of Common carp (Cyprinus Carpio L., 1758; Pisces, Cyprinidae),” American Journal of Environmental Sciences, vol. 5 no.3, pp. 209-217, 2009.
 K. B. Tierney, D. H. Baldwin, T. J. Hara, P. S. Ross, N. L. Scholz, C. J. Kennedy, “Olfactory toxicity in fishes,” Aquatic Toxicology, vol. 96, pp. 2-26, 2010.
 T. J. Hara, “Role of olfaction in fish behaviour,”The Behaviour of Teleost Fishes, T. J. Pitcher, Ed. Springer, 1986, pp. 152-176.
 K. Håkan Olsén, “Effects of pollutants on olfactory detection and responses to chemical cues including pheromones in fish,” Fish pheromones and related cues, P. W. Sorensen, B. D. Wisenden, Ed. Wiley Blackwell, 2015, pp. 217-236.
 S. Budi, B. A. Suliasih, M. S. Othman, L Y. Heng, S. Surif, “Toxicity indetification evaluation of landfill leachate using fish, pawn and seed plant,” Waste Management, to be published.
 Z. B. Salem, N. Capelli, E. Grisey, P. E. Baurand, H. Ayadi, L. Aleya, “First evidence of fish genotoxicity induced by heavy metals from landfill leachates: The advantage of using the RAPD-PCR technique,” Ecotoxicology and Environmental Safety, vol. 101, pp. 90-96, March 2014.
 D. J. Lawrence Thomas, S. F. Tyrrel, R. Smith, S. Farrow, “Biossays for the evaluation onf landfill leachate toxicity,” Journal of Toxicology and Environmental Health, Part B: Critical Reviews, vol. 12, no. 1 pp. 83- 105, 2009.
 M. Zaheer Khan, F. C. P. Law, “Adverse effects of pesticides and related chemicals on enzyme and hormone systems of fish, amphibians and reptiles: a review,” Proceedings of the Pakistan Academy of Sciences, vol. 42, no. 4, pp. 315-323, 2005.
 J. K. Beaulieu, “The Melaleuca Wellness Guide” CO: Littleton, RM Barry Publications, ebook edition, 2015, ch. 3.
 B. A. Flerov, Ecological and physiological aspects of toxicology in fresh-water animals,“ Nauka, Leningrad, pp. 98-104, 1989, (in Russian).
 P. M. Chapman, “Whole effluent toxicity testing – usefulness, level of protection, and risk assessment,” Environmental Toxicology and Chemistry, vol. 19, no. 1, pp. 3-13, January 2000.
 E. Scherer, „Behavioural responses as indicators of environmental alterations: approaches, results, developments,“ Journal of Applied Ichthyology, vol. 8, pp. 122-131, 1992.
 G. Svecevičius, “Behavioral responses of rainbow trout Oncorhynchus mykiss to sublethal toxicity of a model mixture of heavy metals,” Bulletin of Environmental Contamination and Toxicology, vol. 74, pp. 845-852, February 2005.
 G. Svecevičius, “Avoidance response of rainbow trout Oncorhynchus mykiss to hexavalent chromium solutions,“ Bulletin of Environmental Contamination and Toxicology, vol. 79, pp. 596-600, July 2007.
 P. Kavitha, J. Venkateswara Rao, “Oxidative stress and locomotor behaviour response as biomarkers for assessing recovery status of mosquito fish, Gambusia affinis after lethal effect of an organophosphate pesticide, monocrotophos,” Pesticide Biochemistry and Physiology, vol. 87, pp. 182-188, February 2007.
 G. Svecevičius, “Use of behavioral responses of rainbow trout Oncorhynchus mykiss in identifying sublethal exposure to hexavalent chromium,” Bulletin of Environmental Contamination and Toxicology, vol. 82, pp. 564-568, February 2009.
 B. L. Eissa, N. A. Ossana, L. Ferrari, A. Salibián, “Quantative behavioral parameters as toxicity biomarkers: fish responses to waterborne cadmium,” Archives of Environmental Contamination and Toxicology, vol. 58, no. 4, pp. 1032-1039, May 2010.
 C. Vogl, B. Grillitsch, R. Wytek, O. Hunrich Spieser, W. Scholz, “Qualification of spontaneous undirected locomotor behavior of fish for sublethal toxicity testing. Part I. Variability of measurement parameters under general test conditions,” Environmental Toxicology and Chemistry, vol. 18, no. 12, pp. 2736-2742, December 1999.
 A. S. Kane, J. D. Salierno, S. K. Brewer, “Fish models in behavioral toxicology: automated techniques, updates and perspectives,” In: Ostrander GK, Ed. Methods in aquatic toxicology, vol. 2, Lewis Publishers, Boca Raton, FL, pp. 559–590, 2005.
 C. K. Minns, J. R. M. Kelso, R. G. Randall, “Detecting the response of fish to habitat alterations in freshwater ecosystems,” Canadian Journal of Fisheries and Aquatic Sciences, vol. 53, no. S1, pp. 403-414, April 2011.
 E. E. Little, R. D. Archeski, B. A. Flerov, V. I. Kozlovskaya, “Behavioral indicators of sublethal toxicity in rainbow trout,” Archives of Environmental Contamination and Toxicology, vol. 19, pp. 380-385, 1990.
 J. Hellou, K. Cheeseman, E. Desnoyers, D. Johnston, M. L. Jouvenelle, J. Leonard, S. Robertson, P. Walker, “A non-lethal chemically based approach to investigate the quality of harbor sediments,” Science of the Total Environment, vol. 389, no. 1, pp. 178-187, January 2008.
 P. D. Robinson, “Behavioural toxicity of organic chemical contaminants in fish: application to ecological risk assessments (ERAs),” Canadian Journal of Fisheries and Aquatic Sciences, vol. 66, no.7, pp. 1179-1188, July 2009.
 J. Hellou, “Behavioural ecotoxicology, an “early warning” signal to assess environment quality,” Environmental Science and Pollution Research, vol. 18, pp. 1-11, 2011.
 ASTM E1711–12, “Standard guide for measurement of behavior during fish toxicity tests,“ ASTM International, West Conshohocken, PA, pp. 1-15, 2012, doi:10.1520/E1711-12.
 ASTM E1768-95, “Standard guide for ventilator behavioral toxicology testing of freshwater fish,” ASTM International, West Conshohocken, PA, pp. 1-10, 2013, doi: 10.1520/E1768-95R13.
 ASTM E1604-12, “Standard Guide for Behavioral Testing in Aquatic Toxicology,” ASTM International, West Conshohocken, PA, pp. 1-17, 2012, doi: 10.1520/E1604-12
 L. P. J. J. Noldus, A. J. Sink, R. A. J. Tegelenbosch, “EthoVision: a versatile video tracking system for automation of behavioral experiments,” Behavior Research Methods, Instruments and Computers, vol. 33, no. 3, pp. 398-414, 2001.
 K. Suzuki, T. Takagi, T. Hiraishi, “Video analysis of fish schooling behavior infinite space using a mathematical model,” Fisheries Research, vol. 60, pp. 3-10, 2003.
 A. S. Kane, J. D. Salierno, G. T. Gipson, T. C. A. Molteno, C. Hunter, “A video-based movement analysis system to quantify behavioral stress responses of fish,” Water Research, vol. 38, pp. 3993-4001, 2004.
 L. H. Stien, S. Brafland, I. Austevollb, F. Oppedala, T. S. Kristiansen, “A video analysis procedure for assessing vertical fish distribution in aquaculture tanks,” Aquatic Engineering, vol. 37, pp. 115-124, 2007.
 S. Duarte, L. Reig, L. Oca, “Measurement of sole activity by digital image analysis,” Aquatic Engineering, vol. 41, pp. 22-27, 2009.
 V. M. Papadakis, I. E. Papadakis, F. Lamprianidou, A. Glaropoulos, M. Kentouri, “A computer-vision system and methodology for the analysis of fish behavior, “Aquacultural Engineering, 46, pp. 53-59, 2012.
 V. M. Papadakis, A. Glaropoulos, M. Kentouri, “Sub-second analysis of fish behavior using a novel computer-vision system,” Aquacultural Engineering, 62, pp. 36-41, 2014.
 K. J. Buhl, S. J. Hamilton, “Relative sensitivity of early stages of artic grayling, coho salmon, and rainbow trout to nine inorganics,” Ecotoxicology and Environmental Safety, vol. 22, no. 2, pp. 184-197, October 1991.
 G. D. Boeck, W. Meeus, W. De Coen, R. Blust, “Tissue-specific Cu bioaccumulation patterns and differences in sensitivity to waterborne Cu in three freshwater fish: rainbow trout (Oncorhynchus mykiss), common carp (Cyprinus carpio), and gibel carp (Carassius auratus gibelio),” Aquatic Toxicology, vol. 70, no. 3, pp. 179-188, December 2004.
 ISO 10304-1:2007, “Water quality – Determination of dissolved anions by liquid chromatography of ions – Part 1: Determination of bromide, chloride, fluoride, nitrate, nitrite, phosphate and sulfate,” ISO, the International Organization for Standardization.
 ISO 9963-1:1994, “Water quality – Determination of alkalinity – Part 1: Determination of total and composite alkalinity ISO,” the International Organization for Standardization.
 ISO 14911:1998, “Water quality – Determination of disolved Li+, Na+, NH4 +, K+, Mn2+, Ca2+, Mg2+ and Ba2+ using ion chromatography – Method for water and waste water,” ISO, the International Organization for Standardization.
 ISO 10523:2008, “Water quality – Determination of pH,” ISO, the International Organization for Standardization.
 ISO 8467:1993, “Water quality – Determination of permanganate index,” ISO, the International Organization for Standardization.
 ISO 7888:1985, “Water quality – Determination of electrical conductivity,” ISO, the International Organization for Standardization.
 ISO 15586:2003. Water quality – Determination of trace elements using atomic absorption spectrometry with graphite furnace,” ISO, the International Organization for Standardization.
 ISO 12846:2012, “Water quality – Determination of mercury – Method using atomic absorption spectrometry (AAS) with and without enrichment,” ISO, the International Organization for Standardization
 E. Kybartaitė, N. Kazlauskienė, “Toxic effect of the Kairiai landfill leachate on biological parameters of rainbow trout juveniles,” Students scientific workshop, Conference proceedings part II, Vilnius, pp. 37-39, (in Lithuanian).
 C. J. Van Leeuwen, J. L. M. Hermens, “Risk assessment of chemicals: an introduction,” C. J. Leeuwen, T. G. van, Vermeire, Ed. Kluwer Academic Publishers, Dordrecht, 1995.
 J. B. Sprague, “Measurement of pollutant toxicity to fish. III. Sublethal effects and “safe” concentrations,” Water Research, vol. 5, no. 6, pp. 245-266, June 1971.
 G. M. Rand, “Behavior,” Fundamentals of Aquatic Toxicology: Methods and Applications, G. M. Rand and S. R. Petrocelli, Ed., New York: Hemisphere Publishing Co., pp. 221-263.
 T. L. Beitinger, “Behavioral reactions for the assessment of stress in fishes,” Journal of Great Lake Research, vol. 16, no. 4, pp. 495-528, 1990.
 A. B. Barton, „Stress in Fishes: A Diversity of Responses with Particular Reference to Changes in Circulating Corticosteroids,“ Integrative and Comparative Biology, vol. 42, pp. 517-525, 2002.
 G. R. Scott, K. A. Sloman, “The effects of environmental pollutants on complex fish behavior: integrating behavioural and physiological indicators of toxicity: a review,” Aquatic Toxicology, vol. 68, pp. 369- 392, March 2004.
 R. Dhawan, D. B. Dussenbery, P. L. Williams, “Comparison of lethality, reproduction and behavior as toxicological endpoints in the nematode Caenorhabditis elegans,” Journal of Toxicology and Environmental Health, vol. 58, no. 7, pp. 451-462, 1999.
 J. Cairns Jr., W. H. van der Schalie, “Biological monitoring part I–early warning systems,” Water Research, vol. 14, no. 9, pp. 1179-1196, 1980.
 K. J. M. Kramer, J. Botterweg, “Aquatic biological early warning systems: an overview,” in Bioindicators and Environmental Management, D. W. Jeffrey, B. Madden, Ed. London: Academic Press, 1991, pp. 95-126.
 W. H. van der Schalie, T. R. Shedd, P. L. Knechtges, M. W. Widder, “Using higher organisms in biological early warning systems for realtime toxicity detection,” Biosensors and Bioelectronics, vol. 16, no. 7-8, pp. 457-465, September 2001.
 M. J. Bae, Y. S. Park, “Biological early warning system based on the responses of aquatic organisms to disturbances: A review,” Science of the Total Environment, vol. 466-467, pp. 635-649, 2014.
 L. O. Teles, M. Fernandes, J. Amorim, V. Vasconcelos, “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),” Aquatic Toxicology, vol. 165, pp. 241- 248, August 2015.