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Optimizing and Evaluating the Antihelminthic Activity of the Biocompatible Zinc Oxide Nanoparticles Against the Ascaridid Nematode, Parascaris equorum In Vitro

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Abstract

Purpose

In the present study, the effect of different biocompatible concentrations from ZnO nanoparticles (ZnO NPs) on the physiological state and surface topography of the nematode P. equorum was determined in vitro.

Methods

Different concentrations of ZnO NPs (100, 200, 300 and 400 mg/l) synthesized using the egg white were prepared followed by the incubation of parasitic worms with these concentrations in vitro. The physiological state of treated worms such as oxidative stress markers, enzymatic activities and biochemical parameters in addition to the surface topography was determined and compared with control untreated worms.

Results

In comparison to control worms, it was observed that at high concentrations of ZnO NPs, most of the treated worms showed an increase in the levels of ALT, AST and ALP (worm muscle damage, and gonad injury); enhancement of the total protein content (worm cellular dysfunction); significant increase in MDA level (free radical-mediated worm cell membrane damage); depletion in GST and GSH activities (reduced ability to clear toxic compounds like lipid peroxides); CAT depletion (superoxide dismutase and hydrogen peroxide toxicity) and NO increase (detoxification activity and stressful conditions on worms). SEM showed that there was a modified morphological appearance in the surface of treated worms; lips were wrinkled with irregularly arranged denticles, weathering of cuticle, bursts of cuticle layers, disruption of surface annulations and erosion of surface papillae of male around the cloacal opening.

Conclusion

ZnO NPs at environmentally relevant concentrations achieved a significant antihelminthic activity against P. equorum which represents a successful model used in parasite control experiments.

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References

  1. Agarwal H, Kumar V, Shanmugam R (2017) A review on green synthesis of zinc oxide nanoparticles: an eco-friendly approach. Res Eff Technol 3(4):406–413. https://doi.org/10.1016/j.reffit.2017.03.002

    Article  Google Scholar 

  2. Ali D, Alarifi S, Kumar S, Ahamed M, Siddiqui MA (2012) Oxidative stress and genotoxic effect of zinc oxide nanoparticles in freshwater snail Lymnaea luteola L. Aquat Toxicol 124–125:83–90. https://doi.org/10.1016/j.aquatox.2012.07.012

    Article  CAS  PubMed  Google Scholar 

  3. Alves MM, Andrade SM, Grenho L, Fernandes MH, Santos C, Montemor MF (2019) Influence of apple phytochemicals in ZnO nanoparticles formation, photoluminescence and biocompatibility for biomedical applications. Mater Sci Eng C Mater Biol Appl 101:76–87. https://doi.org/10.1016/j.msec.2019.03.084

    Article  CAS  PubMed  Google Scholar 

  4. Amni F, Farahnak A, Golmohammadi T, Eshraghian MR, Rad MBM (2015) The effect of Triclabendazole on ALT enzyme activity in Fasciola hepatica helminths and parasitized sheep liver tissues. J Med Microbiol Infec Dis 3(1–2):1–5

    Google Scholar 

  5. Barrett J, Brophy PM (2000) Ascaris haemoglobin: new tricks for an old protein. Parasitol Today 16(3):90–91

    Article  CAS  Google Scholar 

  6. Bascal ZA, Cunningham JM, Holden-Dye L, O’Shea M, Walker RJ (2001) Characterization of a putative nitric oxide synthase in the neuromuscular system of the parasitic nematode, Ascaris suum. Parasitology 122:219–231

    Article  CAS  Google Scholar 

  7. Becker SL, Liwanag HJ, Snyder JS, Akogun O, Belizario V Jr, Freeman MC, Gyorkos TW, Imtiaz R, Keiser J, Krolewiecki A, Levecke B, Mwandawiro C, Pullan RL, Addiss DG, Utzinger J (2018) Toward the 2020 goal of soil-transmitted helminthiasis control and elimination. PLoS Negl Trop Dis 12(8):e0006606. https://doi.org/10.1371/journal.pntd.0006606

    Article  PubMed  PubMed Central  Google Scholar 

  8. Belfield A, Goldberg DM (1971) Colourimetric determination of alkaline phosphatase activity. Enzyme 12:561–568

    Article  CAS  Google Scholar 

  9. Behnke JM, Buttle DJ, Stepek G, Lowe A, Duce IR (2008) Developing novel anthelmintics from plant cysteine proteinases. Parasit Vectors 1(1):1–29. https://doi.org/10.1186/1756-3305-1-29

    Article  CAS  Google Scholar 

  10. Boomker J, Horak IG, Ramsay KA (1989) Helminth and arthropod parasites of indigenous goats in the Northern Transvaal. Onderstepoort 61:13–20

    Google Scholar 

  11. Brophy PM, Patterson LH, Pritchard DL (1995) Secretory nematode SOD–offensive or defensive? Int J Parasitol 25(7):856–865

    Article  Google Scholar 

  12. Buege JA, Aust SD (1978) Microsomal lipid peroxidation. Methods Enzymol 52:302–310

    Article  CAS  Google Scholar 

  13. Caito SW, Aschner M (2015) Quantification of Glutathione in Caenorhabditis elegans. Curr Protoc Toxicol 64:6–18. https://doi.org/10.1002/0471140856.tx0618s64

    Article  PubMed  PubMed Central  Google Scholar 

  14. Chisholm AD, Xu S (2012) The Caenorhabditis elegans epidermis as a model skin II: differentiation and physiological roles. Wiley Interdiscip Rev Dev Biol 1(6):879–902. https://doi.org/10.1002/wdev.77

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Chitra K, Annadurai G (2013) Antimicrobial activity of wet chemically engineered spherical shaped ZnO nanoparticles on food borne pathogen. Int Food Res J 20(1):59–64

    CAS  Google Scholar 

  16. Chiumiento L, Bruschi F (2009) Enzymatic antioxidant systems in helminth parasites. Parasitol Res 105(3):593–603. https://doi.org/10.1007/s00436-009-1483-0

    Article  PubMed  Google Scholar 

  17. Citiulo F, Jacobsen ID, Miramón P, Schild L, Brunke S, Zipfel P, Brock M, Hube B, Wilson D (2012) Candida albicans scavenges host zinc via Pra1 during endothelial invasion. PLoS Pathog 8:e1002777. https://doi.org/10.1371/journal.ppat.1002777

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Corning S (2009) Equine cyathostomins: a review of biology, clinical significance and therapy. Parasit Vectors 2(Suppl 2):S1. https://doi.org/10.1186/1756-3305-2-S2-S1

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Cox GN, Kusch M, Edgar RS (1981) Cuticle of Caenorhabditis elegans: its isolation and partial characterization. J Cell Biol 90(1):7–17

    Article  CAS  Google Scholar 

  20. Craig TM, Diamond PL, Ferwerda NS, Thompson JA (2007) Evidence of ivermectin resistance by Parascaris equorum on a Texas horse farm. J equine vet sci 27(2):67–71

    Article  Google Scholar 

  21. Danikowski KM, Cheng T (2019) Colorimetric analysis of alkaline phosphatase activity in S. aureus Biofilm. J Vis Exp. https://doi.org/10.3791/59285

    Article  PubMed  Google Scholar 

  22. Dorostkar A, Ghalavand M, Nazarizadeh A, Tat M, Hashemzadeh SM (2017) Anthelmintic effects of zinc oxide and iron oxide nanoparticles against Toxocara vitulorum. Int Nano Lett 7(2):157–164. https://doi.org/10.1007/s40089-016-0198-3

    Article  CAS  Google Scholar 

  23. Doumas BT, Watson WA, Biggs HG (1971) Albumin standards and the measurement of serum albumin with bromcresol green. Clin Chem Acta 31:87–96

    Article  CAS  Google Scholar 

  24. Esmaeilnejad B, Samiei A, Mirzaei Y, Farhang-Pajuh F (2018) Assessment of oxidative/nitrosative stress biomarkers and DNA damage in Haemonchus contortus, following exposure to zinc oxide nanoparticles. Acta Parasitol 63(3):563–571. https://doi.org/10.1515/ap-2018-0065

    Article  CAS  PubMed  Google Scholar 

  25. Fahmy SR, Sayed DA (2017) Toxicological perturbations of zinc oxide nanoparticles in the Coelatura aegyptiaca mussel. Toxicol Ind Health 33(7):564–575. https://doi.org/10.1177/0748233716687927

    Article  CAS  PubMed  Google Scholar 

  26. Gandhi PR, Jayaseelan C, Mary RR, Mathivanan D, Suseem SR (2017) Ascaricidal, pediculicidal and larvicidal activity of synthesized ZnO nanoparticles using Momordica charantia leaf extract against blood feeding parasites. Exp Parasitol 181:47–56. https://doi.org/10.1016/j.exppara.2017.07.007

    Article  CAS  PubMed  Google Scholar 

  27. Gang SS, Hallem EA (2016) Mechanisms of host seeking by parasitic nematodes. Mol Biochem Parasitol 208(1):23–32. https://doi.org/10.1016/j.molbiopara.2016.05.007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Gherbawy YA, Shalaby IM, Abd El–sadek MS, Elhariry HM, Banaja AA (2013) The anti–fasciolasis properties of silver nanoparticles produced by Trichoderma harzianum and their improvement of the anti–fasciolasis drug triclabendazole. Int J Mol Sci 14(11):21887–21898. https://doi.org/10.3390/ijms141121887

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Gonzalez–Moragas L, Roig A, Laromaine A (2015) C. elegans as a tool for in vivo nanoparticle assessment. Adv Colloid Interface Sci 219:10–26. https://doi.org/10.1016/j.cis.2015.02.001

    Article  CAS  PubMed  Google Scholar 

  30. Gopalakrishnan K, Ramesh C, Ragunathan V, Thamilselvan M (2012) Antibacterial activity of Cu2O nanoparticles on E. coli synthesized from Tridax procumbens leaf extract and surface coating with polyaniline. Dig J Nanomater Bios 7:833–839

    Google Scholar 

  31. Habig WH, Pabst MJ, Jakoby WB (1974) Glutathione S–transferases; the first enzymatic step in mercapturic acid formation. J Biol Chem 249:7130–7139

    CAS  Google Scholar 

  32. Henry RJ (1964) Colorimetric determination of total protein. In: clinical chemistry. Harper and Row Publisher, New York, p 181

    Google Scholar 

  33. Hewitson JP, Grainger JR, Maizels RM (2009) Helminth immunoregulation: the role of parasite secreted proteins in modulating host immunity. Mol Biochem Parasitol 167(1):1–11. https://doi.org/10.1016/j.molbiopara.2009.04.008

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Kalpana VN, Devi Rajeswari V (2018) A review on green synthesis, biomedical applications, and toxicity studies of ZnO NPs. Bioinorg Chem Appl 2018:3569758. https://doi.org/10.1155/2018/3569758

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Khan YA, Singh BR, Ullah R, Shoeb M, Naqv AH, Abidi SM (2015) Anthelmintic effect of biocompatible zinc oxide nanoparticles (ZnO NPs) on Gigantocotyle explanatum, a neglected parasite of Indian water buffalo. PLoS One 10(7):e0133086. https://doi.org/10.1371/journal.pone.0133086

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Knox DP, Kennedy MW (1988) Proteinases released by the parasitic larval stages of Ascaris suum, and their inhibition by antibody. Mol Biochem Parasitol 28:207–216

    Article  CAS  Google Scholar 

  37. Kotze AC (2003) Catalase induction protects Haemonchus contortus against hydrogen peroxide in vitro. Int J Parasitol 33:393–400

    Article  CAS  Google Scholar 

  38. Kotze AC, Clifford S, O’Grady J, Behnke JM, McCarthy JS (2004) An in vitro larval motility assay to determine anthelmintic sensitivity for human hookworm and Strongyloides species. Am J Trop Med Hyg 71(5):608–816

    Article  CAS  Google Scholar 

  39. Leslie JF, Cain GD, Meffe GK, Vrijenhoek RC (1982) Enzyme polymorphismin Ascaris suum (Nematoda). J Parasitol 68(4):576–587

    Article  CAS  Google Scholar 

  40. Li YX, Wang Y, Hu YO, Zhong JX, Wang DY (2011) Modulation of the assay system for the sensory integration of 2 sensory stimuli that inhibit each other in nematode Caenorhabditis elegans. Neurosci Bull 27(2):69–82. https://doi.org/10.1007/s12264-011-1152-z

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Liu RM, Gaston Pravia KA (2010) Oxidative stress and glutathione in TGF-β-mediated fibrogenesis. Free Radic Biol Med 48(1):1–15. https://doi.org/10.1016/j.freeradbiomed.2009.09.02

    Article  PubMed  Google Scholar 

  42. Mart´inez A (1995) Nitric oxide synthase in invertebrates. Histochem J 27(10):770–776

    Article  Google Scholar 

  43. Masetti M, Locci T, Cecchettini A, Lucchesi P, Magi M, Malvaldi G, Bruschi F (2004) Nitric oxide synthase immunoreactivity in the nematode Trichinella britovi. Evidence for nitric oxide production by the parasite. Int J Parasitol 34(6):715–721. https://doi.org/10.1016/j.ijpara.2004.02.002

    Article  CAS  PubMed  Google Scholar 

  44. Minning DM, Gow AJ, Bonaventura J, Braun R, Dewhirst M, Goldberg DE, Stamler JS (1999) Ascaris haemoglobin is a nitric oxide-activated deoxygenase. Nature 401(6752):497–502. https://doi.org/10.1038/46822

    Article  CAS  PubMed  Google Scholar 

  45. Morsy K, Bashtar A, Al Quraishy S, Adel S (2016) Description of two equine nematodes, Parascaris equorum Goeze 1782 and Habronema microstoma Schneider 1866 from the domestic horse Equus ferus caballus (Famisly: Equidae) in Egypt. Parasitol Res 115:4299–4306

    Article  Google Scholar 

  46. Murdock RC, Braydich-Stolle L, Schrand AM, Schlager JJ, Hussain SM (2007) Characterization of nanomaterial dispersion in solution prior to in vitro exposure using dynamic light scattering technique. Toxicol Sci 101(2):239–253. https://doi.org/10.1093/toxsci/kfm240

    Article  CAS  PubMed  Google Scholar 

  47. Nandurkar HP, Zambare SP (2012) Comparative study of acute and chronic exposure of chloramphenicol on total lipid contents in different tissues of model animals, Lamellidens corrianus (Lea) and Parreysia cylindrica (Annandale and Prashad). Int J Multidiscip Res 2(3):33–35

    CAS  Google Scholar 

  48. Nazarizadeh A, Asri-Rezaie S (2016) Comparative study of antidiabetic activity and oxidative stress induced by zinc oxide nanoparticles and zinc sulfate in diabetic rats. AAPS Pharm Sci Tech 17(4):834–843. https://doi.org/10.1208/s12249-015-0405-y

    Article  CAS  Google Scholar 

  49. Oberdörster G, Oberdörster E, Oberdörster J (2005) Nanotoxicology: an emerging discipline evolving from studies of ultrafine particles. Environ Health Perspect 113(7):823–839. https://doi.org/10.1289/ehp.7339

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Patterson AL (1939) The Scherrer formula for X-ray particle size determination. Phys Rev 56:978. https://doi.org/10.1103/PhysRev.56.978

    Article  CAS  Google Scholar 

  51. Premanathan M, Karthikeyan K, Jeyasubramanian K, Manivannan G (2011) Selective toxicity of ZnO nanoparticles toward Gram-positive bacteria and cancer cells by apoptosis through lipid peroxidation. Nanomed Nanotechnol Biol Med 7(2):184–192

    Article  CAS  Google Scholar 

  52. Rashid MM, Ferdous J, Banik S, Islam MR, Uddin AH, Robel FN (2016) Anthelmintic activity of silver-extract nanoparticles synthesized from the combination of silvernanoparticles and M. charantia fruit extract. BMC Complement Altern Med 16:242

    Article  Google Scholar 

  53. Reitman S, Frankel S (1957) A colorimetric method for the determination of serum glutamic oxaloacetic and glutamic pyruvic transaminases. Am J Clin Pathol 28(1):56–63

    Article  CAS  Google Scholar 

  54. Richmond W (1973) Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Clin Chem 19(12):1350–1356

    CAS  PubMed  Google Scholar 

  55. Rizvi A, Hasan S, Alam M, Zafar A, Fatima T, Shareef PA, Banu N, Saleemuddin M, Saifullah MK, Abidi SM (2012) Levels of some antioxidant molecules and lipid peroxidation during in vivo transformation of the progenetic metacercaria of Clinostomum complanatum to ovigerous adult worms. Vet Parasitol 185(2–4):164–167. https://doi.org/10.1016/j.vetpar.2011.10.024

    Article  CAS  PubMed  Google Scholar 

  56. Sakr S, Al Lail SM (2005) Fenvalerate induced histopathological and histochemical changes in the liver of the cat fish Clarias gariepinus. J Appl Sci Res 1:263–267

    Google Scholar 

  57. Shafaei S, Al Farahnak, Golmohammadi T, Esharghian MR, Rad MBM (2014) Study of Triclabendazole (TCBZ) effect on Aspartate Transaminase (AST) activity of Fasciola gigantica parasite and liver enzyme activity assay. J Pharma care 2(4):149–153

    Google Scholar 

  58. Shalaby HA (2013) Anthelmintics resistance; how to overcome it? Iran J Parasitol 8(1):18–32

    PubMed  PubMed Central  Google Scholar 

  59. Shoeb M, Singh BR, Khan JA, Khan W, Singh BN, Singh HB, Singh HB, Naqvi AH (2013) ROS-dependent anticandidal activity of zinc oxide nanoparticles synthesized by using egg albumen as a biotemplate. Adv Nat Sci Nanosci Nanotechnol 4:1–11. https://doi.org/10.1088/2043-6262/4/3/035015

    Article  CAS  Google Scholar 

  60. Smith NC, Bryant C (1989) The effects of antioxidants on the rejection of Nippostrongylus brasiliensis. Parasite Immunol 11(2):161–167

    Article  CAS  Google Scholar 

  61. Song S, Guo Y, Zhang X, Zhang X, Zhang J, Ma E (2014) Changes to cuticle surface ultrastructure and some biological functions in the nematode Caenorhabditis elegans exposed to excessive copper. Arch Environ Contam Toxicol 66(3):390–399. https://doi.org/10.1007/s00244-013-9991-4

    Article  CAS  PubMed  Google Scholar 

  62. Starnes D, Unrine J, Chen C, Lichtenberg S, Starnes C, Svendsen C, Kille P, Morgan J, Baddar ZE, Spear A, Bertsch P, Chen KC, Tsyusko O (2019) Toxicogenomic responses of Caenorhabditis elegans to pristine and transformed zinc oxide nanoparticles. Environ Pollut 247:917–926. https://doi.org/10.1016/j.envpol.2019.01.077

    Article  CAS  PubMed  Google Scholar 

  63. Stein EA, Myers G (1994) Lipids, lipoproteins and apolipoproteins. In: Burtis CA, Ashwood ER (eds) Tietz textbook of clinical chemistry. WB Saunders Company, Philadelphia, pp 1002–1993

    Google Scholar 

  64. Tydén E, Dahlberg J, Karlberg O, Höglund J (2014) Deep amplicon sequencing of preselected isolates of Parascaris equorum in β-tubulin codons associated with benzimidazole resistance in other nematodes. Parasit Vectors 29:7–410. https://doi.org/10.1186/1756-3305-7-410

    Article  CAS  Google Scholar 

  65. Wang J, Dai H, Nie Y, Wang M, Yang Z, Cheng L, Liu Y, Chen S, Zhao G, Wu L, Guang S, Xu A (2018) TiO2 nanoparticles enhance bioaccumulation and toxicity of heavy metals in Caenorhabditis elegans via modification of local concentrations during the sedimentation process. Ecotoxicol Environ Saf 162:160–169. https://doi.org/10.1016/j.ecoenv.2018.06.051

    Article  CAS  PubMed  Google Scholar 

  66. Wang N, Wang H, Tang C, Lei S, Shen W, Wang C, Wang G, Wang Z, Wang L (2017) Toxicity evaluation of boron nitride nanospheres and water-soluble boron nitride in Caenorhabditis elegans. Int J Nanomed 8(12):5941–5957. https://doi.org/10.2147/IJN.S130960

    Article  Google Scholar 

  67. Wang Q, Rosa BA, Jasmer DP, Mitreva M (2015) Pan-Nematoda transcriptomic elucidation of essential intestinal functions and therapeutic targets with broad potential. EBioMedicine 2(9):1079–1089. https://doi.org/10.1016/j.ebiom.2015.07.030

    Article  PubMed  PubMed Central  Google Scholar 

  68. Xiong D, Fang T, Yu L, Sima X, Zhu W (2011) Effects of nano-scale TiO2, ZnO and their bulk counterparts on zebrafish: acute toxicity, oxidative stress and oxidative damage. Sci Total Environ 409(8):1444–1452. https://doi.org/10.1016/j.scitotenv.2011.01.015

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

The authors extend their appreciation to the Deanship of the Scientific Research at King Khalid University for funding this work through Research group Project under Grant number (R.G.P.1–56–39).

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Authors and Affiliations

  1. Biology Department, College of Science, King Khalid University, Abha, Saudi Arabia

    Kareem Morsy

  2. Zoology Department, Faculty of Science, Cairo University, Giza, Egypt

    Kareem Morsy, Sohair Fahmy, Ayman Mohamed, Sara Ali, Manal El–Garhy & Mohammed Shazly

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Morsy, K., Fahmy, S., Mohamed, A. et al. Optimizing and Evaluating the Antihelminthic Activity of the Biocompatible Zinc Oxide Nanoparticles Against the Ascaridid Nematode, Parascaris equorum In Vitro. Acta Parasit. 64, 873–886 (2019). https://doi.org/10.2478/s11686-019-00111-2

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