Skip to main content
SpringerLink
Log in

Verapamil caused biochemical alteration, DNA damage, and expression of hepatic stress-related gene biomarkers in Nile tilapia, Oreochromis niloticus

  • Original Article
  • Published:
Comparative Clinical Pathology Aims and scope Submit manuscript

Abstract

Impacts of long-term exposure to sub-lethal verapamil (VRP) at 0.14, 0.28, and 0.57 mg L−1 were examined for 60 days to find the effects on activities of metabolic catalysts as well as molecular reactions in Nile tilapia. Alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase activities in the liver increased significantly (p < 0.05) at high concentration of the drug while there was no significant difference in the activities of soluble phosphatases, corrosive phosphatase, and malate dehydrogenase in comparison with the control. Enzymes of gluconeogenesis, including fructose 1,6-bisphosphatase and glucose-6-phosphatase, were induced while RNA/DNA proportion in fish liver decreased significantly in the fish exposed to the VRP from the 15th day onwards. It caused DNA damage and upregulated expression of hsp70, cat, and sod in the liver, which were found to be correlated with concentration of the drug. Prolonged exposure to sub-lethal VRP could change activities of metabolic catalysts, causing harm to DNA and upregulating stress-related genes in the fish which suggested use of these biomarkers as early cautioning signs about the existence of the drug in aquatic ecosystems.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Afifi M, Alkaladi A, Abu Zinada OA, Couderchet M (2017) Alteration in antioxidant genes expression in some fish caught from Jedda and Yanbu coast as bio-indicator of oil hydrocarbon pollution. Saudi J Biol Sci 24:1580–1587

    CAS  PubMed  Google Scholar 

  • Ajima MNO, Ogo AO, Audu BS, Ugwoegbu KC (2015) Chronic diclofenac (DCF) exposure alters both enzymatic and haematological profile of African catfish, Clarias gariepinus. Drug Chem Toxicol 38:383–390

    CAS  PubMed  Google Scholar 

  • Ajima MNO, Pandey PK, Kundan K, Poojary N, Mane AM (2016) Lipid peroxidation and activities of antioxidant enzymes biomarkers in Oreochromis niloticus (Linnaeus, 1758) chronically treated with diclofenac (DCF). J Exp Zool India 19(2):671–676

  • Ajima MNO, Pandey PK, Kumar K, Poojary N (2017a) Neurotoxic effects, molecular responses and oxidative stress biomarkers in Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) exposed to verapamil. Comp Biochem Physiol C Toxicol Pharmacol 196:44–52

    CAS  PubMed  Google Scholar 

  • Ajima MNO, Pandey PK, Kumar K, Poojary N (2017b) Assessment of mutagenic, heamatological and oxidative stress biomarkers in liver of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) in response to sublethal verapamil exposure. Drug Chem Toxicol 40:286–294

    CAS  PubMed  Google Scholar 

  • Ajima MNO, Pandey PK, Poojary N, Kumar K (2018) Alteration in DNA structure, molecular responses and Na+ K- ATPase activities in the gill of Nile tilapia, Oreochromis niloticus (Linnaeus, 1758) in response to sub-lethal verapamil. Ecotoxicol Environ Saf 147:809–816

    CAS  PubMed  Google Scholar 

  • Akinpelu B, Dare C, Adebesin F, Iwalewa E, Oyedapo O (2013) Effect of stem-bark of Erythrophleum suaveolens (Guill. &Perri.) saponin on fresh water snail (Lanistes lybicus) tissues, Afr. J Environ Sci Technol 6:446–451

    Google Scholar 

  • Al-Rifai JH, Gabefish CL, Schafer AI (2007) Occurrence of pharmaceutically active and non-steroidal estrogenic compounds in three different wastewater recycling schemes in Australia. Chemosphere 69:803–815

    CAS  PubMed  Google Scholar 

  • Anderson D, Yu TW, Philips BJ, Schmerez P (1994) The effects of various antioxidants and other modifying agents on oxygen radical-generated DNA damage in human lymphocytes in comet assay. Mutat Res 307:261–271

    CAS  PubMed  Google Scholar 

  • APHA (American Public Health Association, American Water works Association and Water Environmental Federation) (2005) Standard methods of examination of water and wastewater, 21st edn. APHA, Washington (DC)

    Google Scholar 

  • Asharani PV, Wu YL, Gong Z, Valiyaveettil S (2008) Toxicity of silver nanoparticles in zebrafish models. Nanotechnology 19:255102. https://doi.org/10.1088/09574484/19/25/255102

    Article  CAS  PubMed  Google Scholar 

  • Beiderbeck-Noll AB, Sturkenboom MCJM, Van Der Linden PD, Herings RMC, Hofman A, Coebergh JWW, Leufkens HGM, Stricker BHC (2003) Verapamil is associated with an increased risk of cancer in the elderly: the Rotterdam study. Eur J Cancer 39:98–105

    CAS  PubMed  Google Scholar 

  • Bradford MM (1976) Rapid and sensitive method for quantitation of microgram quantities of protein utilizing principle of protein dye binding. Anal Biochem 72:248–254

    CAS  PubMed  Google Scholar 

  • Brandao FP, Rodrigues S, Castro BB, Gonccalves F, Antunes SC, Nunes B (2013) Short term effects of neuroactive pharmaceutical drugs on a fish species: biochemical and behavioural effects. Aquat Toxicol 144-145:218–229

    CAS  PubMed  Google Scholar 

  • Burkina V (2014) Pharmaceuticals in the aquatic environment and their effects in fish. PhD. Thesis, University of South Bohemia in Ceske Budejovice, Czech Republic, Vodnany.126 pp

  • Chatterjee N, Pal AK, Das T, Dalvi R, Mohammad MS, Sharma K, Mukherjee SC, Baruah K (2010) Effect of stocking density and journey length on the welfare of rohu (Labeo rohita, Hamilton) fry. Aquac Int 18:859–868

    Google Scholar 

  • Essey OA, Lowky OH, Brock MJ (1946) A method for the rapid determination of alkaline phosphatase with five cubic millimeters of serum. J Biol Chem 164:321–329

    Google Scholar 

  • Filby AL, Paull GC, Hickmore TF, Tyler CR (2010) Unraveling the neurophysiological basis of aggression in a fish model. BMC Genomics 11:498–516

    PubMed  PubMed Central  Google Scholar 

  • Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorous. J Biol Chem 66:375–400

    CAS  Google Scholar 

  • Foley CI, Bradley DI, Hook IO (2016) A review of assessment of potential use of RNA: DNA ratios to assess the condition of entrained fish larvae. Ecol Indic 60:346–357

    CAS  Google Scholar 

  • Freeland RA, Harper AL (1959) The study of metabolic pathway by means of adaptation. J Biol Chem 234:1350–1354

    Google Scholar 

  • Jadiya P, Mir SS, Nazir A (2012) Effect of various classes of pesticides on expression of stress genes in transgenic C. elegans model of Parkinson’s disease. CNS Neurol Disord Drug Targets 11:1001–1005

    CAS  PubMed  Google Scholar 

  • James R, Sampath K (1999) Effect of zeolite on the reduction of cadmium toxicity in water and a freshwater fish, Oreochromis mossambicus. Bull Environ Contam Toxicol 62:222–229

    CAS  PubMed  Google Scholar 

  • Khan SJ, Ongerth JE (2004) Modelling of pharmaceutical residues in Australian sewage by quantities of use and fugacity calculations. Chemosphere 54:355–367

    CAS  PubMed  Google Scholar 

  • Khan S, Pandya KP (1985) Hepatotoxicity in albino rats exposed to n-octane and n-nonane. J Appl Toxicol 5:64–68

    CAS  PubMed  Google Scholar 

  • Khan S, Sharma N (2012) A study on enzymes acid phosphatase and alkaline phosphatase in the liver and kidney of fish Gambusia affinis exposed to the chlorpyrifos, an organophosphate. Int J Pharm Sci Rev Res 13:88–90

    CAS  Google Scholar 

  • Kurelec B (1995) Inhibition of multixenobiotic resistance mechanism in aquatic organisms: ecotoxic consequences. Sci Total Environ 171:197–204

    CAS  Google Scholar 

  • Kuropat C, Mercaldo-Allen R, Caldarone E, Goldberg R, Phelan B, Thurberg F (2002) Evaluation of RNA concentration as an indicator of growth in young-of-the-year winter flounder Pseudopleuronectes americanus and Tautog tautoga onitis. Mar Ecol Prog Ser 230:265–274

    CAS  Google Scholar 

  • Le TH, Lim ES, Lee SK, Park JS, Kim YH, Min J (2011) Toxicity evaluation of verapamil and tramadol based on toxicity assay and expression patterns of Dhb, Vtg, Arnt, CYP4, and CYP314 in Daphnia magna. Environ Toxicol 26:515–523

    CAS  PubMed  Google Scholar 

  • Lee RF, Steinert S (2003) Use of the single cell gel electrophoresis/comet assay for detecting DNA damage in aquatic (marine and freshwater) animals. Mutat Res 544:43–64

    CAS  PubMed  Google Scholar 

  • Li ZH, Zlabek V, Velisek J, Grabic R, Machova J, Randak T (2009) Physiological condition status and muscle-based biomarkers in rainbow trout (Oncorhynchus mykiss), after long-term exposure to carbamazepine. J Appl Toxicol 3:197–203

    Google Scholar 

  • Li ZH, Velisek J, Zlabek V, Grabic R, Machova J, Li P, Randak T, Kolarova J (2011) Chronic toxicity of verapamil on juvenile rainbow trout (Oncorhynchus mykiss): effects on morphological indices, hematological parameters and antioxidant responses. J Hazard Mater 185:870–880

    CAS  PubMed  Google Scholar 

  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real time quantitative PCR and the 2-ΔΔCt method. Methods 25:402–408

    CAS  PubMed  Google Scholar 

  • Malarvizhi A, Kavitha C, Saravanan M, Ramesh M (2012) Carbamazepine (CBZ) induced enzymatic stress in gill, liver and muscle of a common carp, Cyprinus carpio. J King Saudi Univ Sci 24:179–186

    Google Scholar 

  • Marjoric AS (1964) In: Colowick SP, Kaplan NO (eds) Methods in enzymology. Vol II. Academic Press Inc., New York 541 pp

    Google Scholar 

  • Mazurais D, Darias M, Zambonino-Infante JL, Cahua CL (2011) Transcriptomics for understanding marine fish larval development. Revue Canadienne de Zoologie 89:599–611

    Google Scholar 

  • Mercaldo-Allen R, Kuropat C, Caldarone EM (2006) A model to estimate growth in young-of-the-year tautog, Tautoga onitis based on RNA/DNA ratio and seawater temperature. J Exp Mar Biol Ecol 329:187–195

    CAS  Google Scholar 

  • Metcalfe CD, Koenig BG, Bennie DT, Servos M, Ternes TA, Hirsch R (2003) Occurrence of neutral and acidic drugs in the effluents of Canadian sewage treatment plants. Environ Toxicol Chem 22:2872–2880

    CAS  PubMed  Google Scholar 

  • Moreno I, Pichardo S, Jos A, Gomez-Amores L, Mate A, Vazquez C, Camean A (2005) Antioxidant enzyme activity and lipid peroxidation in liver and kidney of rats exposed to microcystin-LR administered intraperitoneally. Toxicon 45:395–402

    CAS  PubMed  Google Scholar 

  • Nwani CD, Nagpure N, Kumar R, Kushwaha B, Kumar P, Lakra W (2011) Mutagenic and genotoxic assessment of atrazine-based herbicide to freshwater fish Channa punctatus (Bloch) using micronucleus test and single cell gel electrophoresis. Environ Toxicol Pharmacol 31:314–322

    CAS  PubMed  Google Scholar 

  • Ochoa S (1955) Malic dehydrogenase from pig heart. In: Colowick SP, Kaplan NO (eds) Methods in enzymology. Academic Press, Inc., New York, pp 735–739

    Google Scholar 

  • Overturf M, Overturf C, Baxter D, Hala D, Constantine L, Venables B, Huggett D (2012) Early life-stage toxicity of eight pharmaceuticals to the fathead minnow, Pimephales promelas. Arch Environ Contam Toxicol 62:455–464

    CAS  PubMed  Google Scholar 

  • Palanivelu V, Vijayavel K, Ezhilarasibalasubramanians S, Balasubramanian MP (2005) Impact of fertilizer (urea) on oxygen consumption and feeding the freshwater fish Oreochromis mossambicus. Environ Toxicol Pharmacol 19:351–355

    CAS  PubMed  Google Scholar 

  • Rocco L, Stingo V, Frenzilli G, Zito G, Archimandritis A, Peluso C (2012) Genotoxic effects in fish induced by pharmaceutical agents present in sewage of some Italian water- treatment plants. Environ Toxicol 27:18–25

    CAS  PubMed  Google Scholar 

  • Singh NP, McCoy MT, Tice RR, Schneider EL (1988) A simple technique for quantification of low levels of DNA damage in individual cells. Exp Cell Res 175:184–191

    CAS  PubMed  Google Scholar 

  • Singh N, Rani P, Gupta M, Goel N, Tandan N (2013) Effects of aqueous extract of Camellia sinensis (l.) O. kuntze on liver markers of cadmium treated rats. J Biotechnol Pharm Res 4:89–93

    Google Scholar 

  • Steinbach C, Fedorova G, Prokes M, Grabicova K, Machova J, Grabic R, Valentova O, Kroupova HK (2013) Toxic effects, bioconcentration and depuration of verapamil in the early life stages of common carp (Cyprinus carpio L.). Sci Total Environ 461-462:198–206

    CAS  PubMed  Google Scholar 

  • Stevens JF, Tsang W, Newall RG (1983) Measurement ofthe enzymes lactase dehydrogenase and creatine kinase using relectance spectroscopy and reagent strips. J Clin Pathol 36:1371–1376

    CAS  PubMed  PubMed Central  Google Scholar 

  • Trautwein C, Kummerer K, Metzger JW (2008) Aerobic biodegradability of the calcium channel antagonist verapamil and identification of a microbial dead end transformation product studied by LC–MS/MS. Chemosphere 72:442–450

    CAS  PubMed  Google Scholar 

  • Van Oosten-Hawle P, Porter RS, Morimoto RI (2013) Regulation of organismal protein stasis by transcellular chaperone signaling. Cell 153:1366–1378

    PubMed  PubMed Central  Google Scholar 

  • Varo I, Navarro J, Nunes B, Guilhermino L (2007) Effects of dichlorvos aquaculture treatments on selected biomarkers of gilthead sea bream (Sparus aurata L.) fingerlings. Aquaculture 266:87–96

    CAS  Google Scholar 

  • Velma V, Tchounwou PB (2010) Chromium-induced biochemical, genotoxic and histopathologic effects in liver and kidney of goldfish, Carassius auratus. Mutat Res 698:43–51

    CAS  PubMed  PubMed Central  Google Scholar 

  • Verma AK, Pal AK, Manush SM, Das T, Dalvi RS, Chandrachoodan PP, Ravi PM, Apte SK (2007) Persistent sub-lethal chlorine exposure elicits the temperature, induced stress responses in Cyprinus carpio early fingerlings. Pestic Biochem Physiol 87:229–237

    CAS  Google Scholar 

  • Wooten IDP (1964) Micro analysis in medical biochemistry, 4th edn. J and A Churchill Ltd, London, pp 138–140

    Google Scholar 

  • Yeung JWY, Zhou GJ, Leung KMY (2016) Sub-lethal effects of cadmium and copper on RNA/DNA ratio and energy reserves in the green-lipped mussel Perna viridis. Ecotoxicol Environ Saf 132:59–67

    CAS  PubMed  Google Scholar 

  • Zhou JL, Zhang ZL, Banks E, Grover D, Jiang JQ (2009) Pharmaceutical residues in wastewater treatment work effluents and their impact on receiving river water. J Hazard Mater 166:655–661

    CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are thankful to the Director, ICAR-Central Institute of Fisheries Education, Mumbai, India, for providing the equipment and facilities, used to carry out the research.

Funding

The first author received the grant and finance used in the study from the Indian-African fellowship program.

Author information

Authors and Affiliations

  1. Department of Fisheries and Aquaculture Technology, Federal University of Technology, PMB 1526, Owerri, Nigeria

    Malachy N. O. Ajima

  2. College of Fisheries, Central Agricultural University, Lembucherra, Agartala, Tripura, 799210, India

    Pramod K. Pandey

  3. Aquatic Environment and Health Management Division, ICAR- Central Institute of Fisheries Education, Yari Road, Off Panch Marg, Versova, Andheri West, Mumbai, Maharashtra, 400061, India

    Kundan Kumar & Nalini Poojary

  4. Marine Biotechnology Division, Central Fisheries Research Insistute, Ernakulam North P.O., Kochi, 682018, India

    Adnan H. Gora

Authors
  1. Malachy N. O. Ajima
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Pramod K. Pandey
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Kundan Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nalini Poojary
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Adnan H. Gora
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Malachy N. O. Ajima.

Ethics declarations

Ethnical statements

This study was not funded by any organization or body.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and institutional guidelines for the care and use of animals were followed. (The research undertaken complies with the current animal welfare laws in India. The study was approved by the Board of Studies of Fish of the Central Institute of Fisheries Education (Deemed University), Mumbai, India. The care and treatment of animals used in this study were in accordance with the guidelines of the CPCSEA (Committee for the Purpose of Control and Supervision of Experiments on Animals), Ministry of Environment & Forests (Animal Welfare Division), Govt. of India, on the care and use of animals in scientific research.)

This article does not contain any studies with human participants or animals performed by any of the authors.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ajima, M.N.O., Pandey, P.K., Kumar, K. et al. Verapamil caused biochemical alteration, DNA damage, and expression of hepatic stress-related gene biomarkers in Nile tilapia, Oreochromis niloticus. Comp Clin Pathol 29, 135–144 (2020). https://doi.org/10.1007/s00580-019-03041-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00580-019-03041-7

Keywords

Search

Navigation