Alterations of lactate dehydrogenase activity in the skeletal muscles and cardiac tissue of salmonid after disinfecting procedure with chloramine-t

DOI: 10.32900/2312-8402-2021-125-37-46

Tkachenko H.,
Doctor of Biological Sciences,,
Kurhaluk N.,
Doctor of Biological Sciences,,
Institute of Biology and Earth Sciences, Pomeranian University in Słupsk, Poland,
Grudniewska J.,
Ph. D.,
Stanislaw Sakowicz Inland Fisheries Institute, Poland

Keywords: rainbow trout (Oncorhynchus mykiss Walbaum), brown trout (Salmo trutta m. fario), grayling (Thymallus thymallus Linck), muscles, cardiac tissue, lactate dehydrogenase, disinfection


Chloramine-T is a widely used disinfectant for the treatment of gill diseases of fish in freshwater and can be toxic to fish. Therefore, the current study aimed to examine the safety of this disinfecting product (as it has been attracting researchers’ attention for applying in aquatic animals) for fish health using markers of aerobic and anaerobic capacity (i.e. lactate dehydrogenase activity) in the skeletal muscle and cardiac tissues of rainbow trout (Oncorhynchus mykiss Walbaum), grayling (Thymallus thymallus Linck), and brown trout (Salmo trutta m. fario). Twenty-two clinically healthy rainbow trout, twenty-one brown trout, and twenty graylings were exposed to Chloramine-T in a final concentration of 9 mg per L. The Control group of fish was handled in the same way as the exposed groups. Fish were bathed for 20 min and repeated three times every 3 days. Two days after the last bathing fish were sampled. In the skeletal muscle tissue, LDH activity was decreased in the rainbow trout and grayling after disinfection by Chloramine-T compared to the unhandled controls. On the other hand, LDH activity was increased in the skeletal muscle tissue of brown trout. In the cardiac tissue, disinfection by Chloramine-T caused the decrease of LDH activity in rainbow trout, brown trout, and grayling. Moreover, in unhandled controls, LDH activity in the cardiac tissue was higher by 107.5 % (p = 0.000) in brown trout and by 57.6 % (p = 0.001) in the grayling compared to the values obtained in skeletal muscles. The present investigation demonstrates the alterations in LDH activity in the skeletal muscles and cardiac tissue after the disinfecting procedure with Chloramine-T in dose 9 mg per L. Although, after disinfection, the rainbow trout, brown trout, and grayling showed decreased trends of aerobic responses in the cardiac tissue indicating adaptive response against the Chloramine-T toxicity. Similar trends were observed in the skeletal muscles of rainbow trout and grayling. On the other hand, LDH activity in the skeletal muscles of brown trout after the disinfecting procedure with Chloramine-T was increased. Therefore, these biochemical parameters can be considered as indicators for the assessment of disinfecting effects, although further studies are required for investigating the mechanism involved in this pattern.


  1. Arnitz, R., Nagl, M., & Gottardi, W. (2009). Microbicidal activity of monochloramine and chloramine T compared. Hosp. Infect., 73(2), 164–170.
  2. Bills, T. , Marking, L. L., Dawson, V. K., Howe, & G. E. (1988). Effects of organic matter and loading rates of fish on the toxicity of chloramine-T. Investigations in Fish Control Report 97. U.S. Fish and Wildlife Service. Available from the Publications Unit, U.S. Fish and Wildlife Service, Springfield, Virginia. 4 pp.
  3. Bills, T. , Marking, L. L., Dawson, V. K., & Rach, J. J. (1988). Effects of environmental factors on the toxicity of chloramine-T to fish. Investigations in Fish Control Report 96. U.S. Fish and Wildlife Service. Available from the Publications Unit, U.S. Fish and Wildlife Service, Springfield, Virginia. 6 pp.
  4. Bills, T. , Marking, L. L., & Howe, G. E. (1993). Sensitivity of juvenile striped bass to chemicals used in aquaculture. U.S. Fish and Wildlife Service Technical Report Series 192. Available from the Publications Unit, U.S. Fish and Wildlife Service, Springfield, Virginia. 11 pp.
  5. Bootsma, R. (1973). Infections with Saprolegnia in pike culture (Esox lucius). Aquaculture, 2, 385–394.
  6. Bradford, M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248–254.
  7. Chloramine-T [127-65-1] and Metabolite p-Toluenesulfonamide [70-55-3]. Review of Toxicological Literature. Prepared for Scott Masten, Ph.D., National Institute of Environmental Health Sciences, Submitted by Karen E. Haneke, M.S. Integrated Laboratory Systems, 2002.
  8. Churova, M. , Meshcheryakova, O. V., Veselov, A. E., Efremov, D. A., & Nemova, N. N. (2017). Activity of metabolic enzymes and muscle-specific gene expression in parr and smolts Atlantic salmon Salmo salar L. of different age groups. Fish Physiol. Biochem., 43 (4), 1117–1130.
  9. Cross, D.G., Hursey, P.A. (1973). Chloramine-T for the control of Ichthyophthirius multifiliis (Fouquet). Journal of Fish Biology, 5, 789–798.
  10. Elia, A. , Magara, G., Righetti, M., Dörr, A. J., Scanzio, T., Pacini, N., Abete, M. C., & Prearo, M. (2017). Oxidative stress and related biomarkers in cupric and cuprous chloride-treated rainbow trout. Environ. Sci. Pollut. Res. Int., 24 (11), 10205–10219.
  11. Ferreira, G. S., Rosalen, P. L., Peixoto, L. R., Pérez, A. L. A. L., Carlo,F. G. C., Castellano, L. R. C., Lima, J. M., Freires, I. A., Lima, E. O., & Castro, R. D. (2017). Antibiofilm Activity and Mechanism of Action of the Disinfectant Chloramine-T on Candida spp., and Its Toxicity against Human Cells. Molecules, 22(9), 1527.
  12. Gottardi, W., Debabov, D., & Nagl, M. (2013). N-chloramines, a promising class of well-tolerated topical anti-infectives. Agents Chemother., 57 (3), 1107–1114.
  13. Isaac, R. , Morris, J. C. (1983). Transfer of active chlorine from chloramine to nitrogenous organic compounds. 1. Kinetics. Environ. Sci. Technol., 17 (12), 738–742.
  14. Mattice, J. , Tsai, S. C. (1983). Total residual chlorine as a regulatory tool. In: R.L. Jolley et al., editors. Water chlorination: Environmental impact and health effects. Vol 4. Ann. Arbor. Science Publishers, Ann Arbor, Michigan. P. 901-912.
  15. Orrego, R., Pandelides, Z., Guchardi, J., & Holdway, D. (2011). Effects of pulp and paper mill effluent extracts on liver anaerobic and aerobic metabolic enzymes in rainbow trout. Environ. Saf., 74(4), 761–768.
  16. Powell, M. , Perry, S. F. (1996). Respiratory and acid-base disturbances in rainbow trout (Oncorhynchus mykiss) blood during exposure to chloramine-T, para-toluenesulfonamide, and hypochlorite. Canadian Journal of Fisheries and Aquatic Sciences, 53, 701–708.
  17. Schmidt, L. , Gaikowski, M. P., Gingerich, W. H., Stehly, G. R., Larson,W. J., Dawson, V. K., & Schreier, T. M. (2007). Environmental Assessment of the Effects of Chloramine-T Use in and Discharge by Freshwater Aquaculture. Submitted to U.S. Food and Drug Administration Center for Veterinary Medicine Director, Division of Therapeutic Drugs for Food Animals Office of New Animal Drug Evaluation, Maryland, USA, 136 p.
  18. Sevela, M., Tovarek, J. (1959). Metoda stanovení laktikodehydrogenázy v télních tekutinách [Method for the estimation of lactic dehydrogenase]. Lek. Cesk., 98(27), 844–848. Czech.
  19. Somero, G. , Childress, J. J. (1980). A violation of the metabolism-size scaling paradigm: activities of glycolytic enzymes in muscle increase in larger size fish. Physiol. Zool., 53, 322–337.
  20. Tkachenko, G. , Grudniewska, J. (2015). Tissue-specific response of protein oxidation in the grayling (Thymallus thymallus L.) disinfected by chloramine-T. Scientific Medical Bulletin, 1(1), 76–82.
  21. Tkachenko, H., Grudniewska, J. (2016a). Biochemical changes in the muscle tissue of rainbow trout (Oncorhynchus mykiss Walbaum) disinfected by Chloramine-T. Baltic Coastal Zone – Journal of Ecology and Protection of the Coastline, 20, 101–116.
  22. Tkachenko, H., Grudniewska, J. (2016b). Influence of chloramine-T on oxidative stress biomarkers in the muscle tissue of grayling (Thymallus thymallus). Scientific journal «Kaliningrad State Technical University News», 42, 49–58.
  23. Tkachenko, H., Grudniewska, J. (2016c). Influence of chloramine-T on oxidative stress biomarkers in the cardiac tissue of grayling (Thymallus thymallus). In: Globalisation and regional environment protection. Technique, technology, ecology. Eds T. Noch, W. Mikołajczewska, A. Wesołowska. Gdańsk, Gdańsk High School Publ., 2016. – P. 213–234.
  24. Tkachenko, H., Grudniewska, J. (2016d). Lipid and protein oxidation in the muscle tissue of grayling (Thymallus thymallus) after Chloramine-T disinfection. Proceedings of the International Forum “The Current State and Prospects for the Development of Aquaculture in the Caspian Region“, dedicated to the 85th anniversary of Dagestan State University and the 75th anniversary of Professor F. Magomayev. Ed. F. Magomayev, S. Chalayeva, S. Kurbanova, A. Shakhnazova (Makhachkala, 17–19 October, 2016). Makhachkala, Printing house IPE RD, 2016. p. 168–175.
  25. Tkachenko, H., Grudniewska, J. (2016e). Effect of chloramine-T disinfection on oxidative stress biomarkers in the gill tissue of grayling (Thymallus thymallus). Trudy VNIRO, 162, 150–160.
  26. Tkachenko, H., Grudniewska, J. (2016f). Oxidative stress biomarkers in the muscle tissue of grayling (Thymallus thymallus Linck) after Chloramine-T disinfection. Słupskie Prace Biologiczne, 13, 231–252.
  27. Tkachenko, H., Kurhaluk, N., Grudniewska, J. (2012). Effects of Chloramine-T exposure on oxidative stress biomarkers and liver biochemistry of rainbow trout, Oncorhynchus mykiss (Walbaum), brown trout, Salmo trutta (L.), and grayling, Thymallus thymallus. Pol. Fish., 21, 41–51.
  28. Zar, J. (1999). Biostatistical Analysis, 4th ed., Prentice Hall Inc., New Jersey.