Advertisement

Laboratory bioassays on the response of honey bee (Apis mellifera L.) glutathione S-transferase and acetylcholinesterase to the oral exposure to copper, cadmium, and lead

  • Tatjana V. NikolićEmail author
  • Danijela Kojić
  • Snežana Orčić
  • Elvira L. Vukašinović
  • Duško P. Blagojević
  • Jelena Purać
Research Article
  • 48 Downloads

Abstract

In the present study, the influence of cadmium, copper, and lead on two enzymes often used as biomarkers in toxicological analysis was investigated. Bees were fed with 1 M sucrose solution containing 10-fold serial dilutions of CuCl2 (1000 mg L−1, 100 mg L−1, and 10 mg L−1), CdCl2 (0.1 mg L−1, 0.01 mg L−1, and 0.001 mg L−1), or PbCl2 (10 mg L−1, 1 mg L−1, and 0.1 mg L−1) during 48 h. Our results showed that the total glutathione S-transferase activity was not changed under the influence of cadmium and lead, and it was decreased with the highest concentration of copper. The level of gene expression of the three analyzed classes of glutathione S-transferase was significantly increased with increasing concentrations of copper and cadmium. Lead did not cause significant changes in glutathione S-transferase activity and gene expression, while it showed biphasic effect on acetylcholinesterase activity: lower concentration of lead, 0.1 mg L−1 inhibited and higher dose, 10 mg L−1 induced acetylcholinesterase activity in honey bees. Furthermore, our results showed a significant decrease of the acetylcholinesterase activity in honey bees treated with 0.001 and 0.01 mg L−1 CdCl2. Our results indicate the influence of cadmium, copper, and lead on GST and AChE in the honey bees. These results form the basis for future research on the impact of metallic trace element pollution on honey bees.

Keywords

Honey bee Laboratory tests Metallic trace elements Detoxification Gene expression Enzyme activity 

Notes

Funding information

This work was funded by the Ministry of Education, Science and Technological Development of the Republic of Serbia, Grant no. 173014, project entitled “Molecular mechanisms of redox signalling in homeostasis: adaptation and pathology.”

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Ahmad S (1995) Oxidative stress from environmental pollutants. Arch Insect Biochem Physiol 29:135–157CrossRefGoogle Scholar
  2. Antúnez K, Martín-Hernández R, Prieto L, Meana A, Zunino P, Higes M (2009) Immune suppression in the honey bee (Apis mellifera) following infection by Nosema ceranae (Microsporidia). Environ Microbiol 11:2284–2290CrossRefGoogle Scholar
  3. Aufauvre J, Misme-Aucouturier VB, Texier C, Delbac F, Blot N (2014) Transcriptome analyses of the honey bee response to Nosema ceranae and insecticides. PLoS One 9(3):e91686CrossRefGoogle Scholar
  4. Badiou A, Belzunces LP (2008) Is acetylcholinesterase a pertinent biomarker to detect exposure of pyrethroids? A study case with deltamethrin. Chem Biol Interact 175:406–409CrossRefGoogle Scholar
  5. Badiou-Beneteau A, Carvalho SM, Brunet J, Carvalho GA, Bulete A, Giroud B, Belzunces LP (2012) Development of biomarkers of exposure to xenobiotics in the honey bee Apis mellifera: application to the systemic insecticide thiamethoxam. Ecotoxicol Environ Saf 82:22–31CrossRefGoogle Scholar
  6. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  7. Brown RJ, Galloway TS, Lowe D, Browne MA, Dissanayake A, Jones MB, Depledge MH (2004) Differential sensitivity of three marine invertebrates to copper assessed using multiple biomarkers. Aquat Toxicol 66:267–278CrossRefGoogle Scholar
  8. Buchwalter DB (2008) Metals. In: Smart RC, Hodgson E (eds) Molecular and biochemical toxicology. John Wiley & Sons, Inc., Hoboken, pp 413–439CrossRefGoogle Scholar
  9. Calisi A, Zaccarelli N, Lionetto MG, Schettino T (2013) Integrated biomarker analysis in the earthworm Lumbricus terrestris: application to the monitoring of soil heavy metal pollution. Chemosphere 904:2637–2644CrossRefGoogle Scholar
  10. Celli G, Maccagnani B (2003) Honey bees as bioindicators of environmental pollution. Bull Insectology 56:137–139Google Scholar
  11. Claudianos C, Ranson H, Johnson RM, Biswas S, Schuler MA, Berenbaum MR, Feyereisen R, Oakeshott JG (2006) A deficit of detoxification enzymes: pesticide sensitivity and environmental response in the honeybee. Insect Mol Biol 15:615–636CrossRefGoogle Scholar
  12. Collet C, Belzunces L (2007) Excitable properties of adult skeletal muscle fibres from the honeybee Apis mellifera. J Exp Biol 210:454–464CrossRefGoogle Scholar
  13. Čolović MB, Krstić DZ, Lazarević-Pašti TD, Bondžić AM, Vasić VM (2013) Acetylcholinesterase inhibitors: pharmacology and toxicology. Curr Neuropharmacol 11:315–335CrossRefGoogle Scholar
  14. Conti ME, Botre F (2001) Honeybees and their products as potential bioindicators of heavy metals contamination. Environ Monit Assess 69:267–282CrossRefGoogle Scholar
  15. Corona M, Robinson GE (2006) Genes of the antioxidant system of the honey bee: annotation and phylogeny. Insect Mol Biol 15:687–701CrossRefGoogle Scholar
  16. Costa JR, Mela M, de Assis HC, Pelletier É, Randi MA, de Oliveira Ribeiro CA (2007) Enzymatic inhibition and morphological changes in Hoplias malabaricus from dietary exposure to lead(II) or methylmercury. Ecotoxicol Environ Saf 67:82–88CrossRefGoogle Scholar
  17. Cunha I, Mangas-Ramirez E, Guilhermino L (2007) Effects of copper and cadmium on cholinesterase and glutathione S-transferase activities of two marine gastropods (Monodonta lineata and Nucella lapillus). Comp Biochem Physiol C 145:648–657Google Scholar
  18. de Lima D, Roque GM, de Almeida EA (2013) In vitro and in vivo inhibition of acetylcholinesterase and carboxylesterase by metals in zebrafish (Danio rerio). Mar Environ Res 91:45–51CrossRefGoogle Scholar
  19. Di N, Hladun KR, Zhang K, Liu T, Trumble JT (2016) Laboratory bioassays on the impact of cadmium, copper and lead on the development and survival of honeybee (Apis mellifera L.) larvae and foragers. Chemosphere 152:530–538CrossRefGoogle Scholar
  20. Ding Y, Ortelli F, Rossiter LC, Hemingway J, Ranson H (2003) The Anopheles gambiae glutathione transferase supergene family: annotation, phylogeny and expression profiles. BMC Genomics 4:35CrossRefGoogle Scholar
  21. Ellman GL, Courtney KD, Andres JV, Featherstone RM (1961) A new rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol 7:88–95CrossRefGoogle Scholar
  22. Ercal N, Gurer-Orhan H, Aykin-Burns N (2001) Toxic metals and oxidative stress part I: mechanisms involved in metal induced oxidative damage. Curr Top Med Chem 1:529–539CrossRefGoogle Scholar
  23. Evans JD, Schwartz RS, Chen YP, Budge G, Cornman RS, DeLaRua P, DeMiranda JR, Foret S, Foster L, Gauthier L, Genersch E, Gisder S, Jarosch A, Kucharski R, Lopez D, Lun CM, Moritz RFA, Maleszka R, Muñoz I, Pinto MA (2013) Standard methodologies for molecular research in Apis mellifera. In: Dietemann, V, Ellis JD, Neumann P (eds), The COLOSS BEEBOOK, Volume I: standard methods for Apis mellifera research. J Apic Res 52.  https://doi.org/10.3896/IBRA.1.52.4.11
  24. Forget J, Pavillon J, Beliaeff B, Bocquene G (1999) Joint action of pollutant combinations (pesticides and metals) on survival (LC50 values) and acetylcholinesterase activity of Tigriopus brevicornis (Copepoda, Harpacticoida). Environ Toxicol Chem 18:912–918CrossRefGoogle Scholar
  25. Formicki G, Greń A, Stawarz R, Zyśk B, Gał A (2013) Metal content in honey, propolis, wax, and bee pollen and implications for metal pollution monitoring. Pol J Environ Stud 22:99–106Google Scholar
  26. Frasco MF, Colletier J, Weik M, Carvalho F, Guilhermino L, Stojan J, Fournier D (2007) Mechanisms of cholinesterase inhibition by inorganic mercury. FEBS J 274:1849–1861CrossRefGoogle Scholar
  27. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S-transferases. J Biol Chem 29:7130–7139Google Scholar
  28. Hayes JD, Flanagan JU, Jowsey IR (2005) Glutathione transferases. Annu Rev Pharmacol 45:51–88CrossRefGoogle Scholar
  29. Hladun KR, Di N, Liu TX, Trumble JT (2016) Metal contaminant accumulation in the hive: consequences for whole-colony health and brood production in the honey bee (Apis mellifera L.). Environ Toxicol Chem 35:322–329CrossRefGoogle Scholar
  30. Jensen CS, Garsdal L, Baatrup E (1997) Acetylcholinesterase inhibition and altered locomotor behavior in the carabid beetle Pterostichus cupreus. A linkage between biomarkers at two levels of biological complexity. Environ Toxicol Chem 16:1727–1732CrossRefGoogle Scholar
  31. Johnson RM (2015) Honey bee toxicology. Annu Rev Entomol 60:415–434CrossRefGoogle Scholar
  32. Kang JS, Lee DW, Koh YH, Lee SH (2011) A soluble acetylcholinesterase provides chemical defense against xenobiotics in the pinewood nematode. PLoS One 6:e19063CrossRefGoogle Scholar
  33. Kim BY, Hui WL, Lee KS, Wan H, Yoon HJ, Gui ZZ, Chen S, Jin BR (2011) Molecular cloning and oxidative stress response of a sigma-class glutathione S-transferase of the bumblebee Bombus ignitus. Comp Biochem Physiol B 158:83–89CrossRefGoogle Scholar
  34. Kim YH, Cha DJ, Jung JW, Kwon HW, Lee SH (2012) Molecular and kinetic properties of two acetylcholinesterases from the western honey bee, Apis mellifera. PLoS One 7:e48838CrossRefGoogle Scholar
  35. Kim YH, Kwon DH, Ahn HM, Koh YH, Lee SH (2014) Induction of soluble AChE expression via alternative splicing by chemical stress in Drosophila melanogaster. Insect Biochem Mol Biol 48:75e82CrossRefGoogle Scholar
  36. Lambert O, Piroux M, Puyo S, Thorin C, Larhantec M, Delbac F, Pouliquen H (2012) Bees, honey and pollen as sentinels for lead environmental contamination. Environ Pollut 170:254–259CrossRefGoogle Scholar
  37. Lee KW, Raisuddin S, Rhee JS, Hwang DS, Yu IT, Lee YM, Park HG, Lee JS (2008) Expression of glutathione S-transferase (GST) genes in the marine copepod Tigriopus japonicus exposed to trace metals. Aquat Toxicol 89:158–166CrossRefGoogle Scholar
  38. Li X (2009) Glutathione and glutathione-S-transferase in detoxification mechanisms. In: Ballantyne B, Marrs TC, Syversen T (eds) General and applied toxicology, third edn, vol 2009. John Wiley & Sons ltd, Chichester, pp 411–423Google Scholar
  39. Lionetto MG, Caricato R, Calisi A, Schettino T (2011) Acetylcholinesterase inhibition as a relevant biomarker in environmental biomonitoring: new insights and perspectives. In: Visser JE (ed) Ecotoxicology around the globe. Nova Science Publishers, New York, pp 87–115Google Scholar
  40. Lionetto MG, Caricato R, Calisi A, Giordano ME, Schettino T (2013) Acetylcholinesterase as a biomarker in environmental and occupational medicine: new insights and future perspectives. Biomed Res Int 2013:1–8CrossRefGoogle Scholar
  41. Lourenço AP, Mackert A, Cristino AD, Simões ZLP (2008) Validation of reference genes for gene expression studies in the honey bee, Apis mellifera, by quantitative real-time RT-PCR. Apidologie 39:372–385CrossRefGoogle Scholar
  42. Morimoto T, Kojima Y, Toki T, Komeda Y, Yoshiyama M, Kimura K, Nirasawa K, Kadowaki T (2011) The habitat disruption induces immune-suppression and oxidative stress in honey bees. Ecol Evol 1:201–217CrossRefGoogle Scholar
  43. Nair PMG, Choi J (2011) Identification, characterization and expression profiles of Chironomus riparius glutathione S-transferase (GST) genes in response to cadmium and silver nanoparticles exposure. Aquat Toxicol 101:550–560CrossRefGoogle Scholar
  44. Negri I, Mavris C, Di Prisco G, Caprio E, Pellecchia M (2015) Honey bees (Apis mellifera, L.) as active samplers of airborne particulate matter. PLoS One 10:e0132491CrossRefGoogle Scholar
  45. Nikolić TV, Kojić D, Orčić S, Batinić D, Vukašinović E, Blagojević DP, Purać J (2016) The impact of sublethal concentrations of copper, lead and cadmium on honey bee redox status, superoxide dismutase and catalase in laboratory conditions. Chemosphere 164:98–105CrossRefGoogle Scholar
  46. Perić-Mataruga V, Petković B, Ilijin L, Mrdaković M, Dronjak Čučaković S, Todorović D, Vlahović M (2017) Cadmium and high temperature effects on brain and behaviour of Lymantria dispar L. caterpillars originating from polluted and less polluted forests. Chemosphere 185:628–636CrossRefGoogle Scholar
  47. Perugini M, Manera M, Grotta L, Abete MC, Tarasco R, Amorena M (2011) Heavy metal (Hg, Cr, Cd, and Pb) contamination in urban areas and wildlife reserves: honey bees as bioindicators. Biol Trace Elem Res 140:170–176CrossRefGoogle Scholar
  48. Qin G, Jia M, Liu T, Zhang X, Guo Y, Zhu KY, Ma E, Zhang J (2013) Characterization and functional analysis of four glutathione S-transferases from the migratory locust, Locusta migratoria. PLoS One 8(3):e58410CrossRefGoogle Scholar
  49. Roman A (2007) Content of some trace elements in fresh honeybee pollen. Pol J Food Nutr Sci 57:475–478Google Scholar
  50. Salazar-Medina AJ, García-Rico L, García-Orozco KD, Valenzuela-Soto E, Contreras-Vergara CA, Arreola R, Arvizu-Flores A, Sotelo-Mundo RR (2010) Inhibition by Cu2+ and Cd2+ of a mu-class glutathione S-transferase from shrimp Litopenaeus vannamei. J Biochem Mol Toxicol 24:218–222CrossRefGoogle Scholar
  51. Sarkar A, Ray D, Shrivastava AN, Sarker S (2006) Molecular biomarkers: their significance and application in marine pollution monitoring. Ecotoxicology 15:333–340CrossRefGoogle Scholar
  52. Schmehl DR, Teal PEA, Frazier JL, Grozinger CM (2014) Genomic analysis of the interaction between pesticide exposure and nutrition in honey bees (Apis mellifera). J Insect Physiol 71:177–190CrossRefGoogle Scholar
  53. Sherratt PJ, Hayes JD (2001) Glutathione S-transferases. In: Ioannides C (ed) Enzyme systems that metabolize drugs and other xenobiotics. John Wiley & Sons, Ltd, Chichester, pp 319–352Google Scholar
  54. Sigh SP, Coronella JA, Beneš H, Cochrane BJ, Zimniak P (2001) Catalytic function of Drosophila melanogaster glutathione S-transferase DmGSTS1-1 (GST-2) in conjugation of lipid peroxidation end products. Eur J Biochem 268:2912–2923CrossRefGoogle Scholar
  55. Stohs SJ, Bagchi D (1995) Oxidative mechanisms in the toxicity of metal ions. Free Radic Biol Med 18:321–336CrossRefGoogle Scholar
  56. Stone D, Jepson P, Laskowski R (2002) Trends in detoxification enzymes and heavy metal accumulation in ground beetles (Coleoptera: Carabidae) inhabiting a gradient of pollution. Comp Biochem Physiol C 132:105–112Google Scholar
  57. The Honeybee Genome Sequencing Consortium (2006) Insights into social insects from the genome of the honeybee Apis mellifera. Nature 443:931–949CrossRefGoogle Scholar
  58. Udomsinprasert R, Pongjaroenkit S, Wongsantichon J, Oakley AJ, Prapanthadara L, Wilce MCJ, Ketterman AJ (2005) Identification, characterization and structure of a new Delta class glutathione transferase isoenzyme. Biochem J 388:763–771CrossRefGoogle Scholar
  59. vanEngelsdorp D, Meixner MD (2010) A historical review of managed honey bee populations in Europe and the United States and the factors that may affect them. J Invertebr Pathol 103:S80–S95CrossRefGoogle Scholar
  60. Wang B, Du Y (2013) Cadmium and its neurotoxic effects. Oxidative Med Cell Longev 2013:898034Google Scholar
  61. Wang S, Shi X (2001) Molecular mechanisms of metal toxicity and carcinogenesis. Mol Cell Biochem 222:3–9CrossRefGoogle Scholar
  62. Yan H, Jia H, Wang Z, Gao H, Guo X, Xu B (2013a) Identification and characterization of an Apis cerana cerana Delta class glutathione S-transferase gene (AccGSTD) in response to thermal stress. Naturwissenschaften 100:153–163CrossRefGoogle Scholar
  63. Yan H, Jia H, Gao H, Guo X, Xu B (2013b) Identification, genomic organization, and oxidative stress response of a sigma class glutathione S-transferase gene (AccGSTS1) in the honey bee, Apis cerana cerana. Cell Stress Chaperones 18:415–426CrossRefGoogle Scholar
  64. Zimniak P, Singh SP (2006) Families of glutathione transferases. In: Awasthi YC (ed) Toxicology of glutathione transferases. CRC press LLC, Boca Raton, pp 777–780Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biology and EcologyUniversity of Novi Sad, Faculty of SciencesNovi SadRepublic of Serbia
  2. 2.Institute for Biological Research “Siniša Stanković”University of BelgradeBelgradeRepublic of Serbia

Personalised recommendations