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Exposure to ethylene thiourea degrades the sperm ability of mammals

  • Received : 2024.02.07
  • Accepted : 2024.03.22
  • Published : 2024.06.01

Abstract

Mancozeb is a manganese and zinc-containing fungicide that belongs to the ethylene bisdithiocarbamate group and produces ethylene thiourea (ETU) after biotransformation or environmental degradation, which has toxicological hazard owing to its known antithyroid properties. Although mancozeb leads to negative changes in fertility capacity, the effects of ETU are less known. Therefore, this study examined the alteration of fertilization competence in boar spermatozoa exposed to ETU. The sperm motility, motion kinematics, viability, acrosome integrity, chromatin stability, and intracellular reactive oxygen species (ROS) production of sperm subjected to various ETU concentrations (10, 50, 100, and 200 µM) were evaluated after two different incubation times (30 min and 2 hrs). In addition, the relative mRNA expression of the sperm functional proteins was analyzed after exposure to ETU. A dose-dependent motility reduction was observed in sperm exposed to ETU during both incubation periods compared to the controls. The motion kinematics were reduced significantly in sperm incubated with ETU. Higher percentages of viable sperm were observed in the controls, while such viability was decreased significantly in sperm with 10 - 200 µM ETU. The acrosome integrity was particularly damaged on sperm incubated with 10 - 200 µM ETU for 30 min. Higher intracellular ROS levels were produced in sperm exposed to 200 µM ETU. In addition, lower relative levels of AKAP3, AKAP4, ODF2, and ZPBP2 expression were observed in sperm exposed to ETU compared to the controls. Mancozeb and ETU could adversely affect the reproductive functions of mammals. Hence, the effects of ETU on the reproductive system should be examined further.

Keywords

Acknowledgement

This work was supported by a Research Promotion Program of SCNU.

References

  1. Adikari AA, Moon ST, Yi YJ. 2022. Deterioration in the fertilization capability of boar spermatozoa upon exposure to mancozeb. Korean Journal of Agricultural Science 49:259-267. https://doi.org/10.7744/KJOAS.20220023
  2. Aktar MW, Paramasivam M, Sengupta D, Purkait S, Ganguly M, Banerjee S. 2009. Impact assessment of pesticide residues in fish of Ganga river around Kolkata in West Bengal. Environmental Monitoring and Assessment 157:97-104. https://doi.org/10.1007/s10661-008-0518-9
  3. Aruldhas MM, Ramalingam N, Jaganathan A, John Sashi AM, Stanley JA, Nagappan AS, Vasavan J, Kannan A, Seshadri VN. 2010. Gestational and neonatal-onset hypothyroidism alters androgen receptor status in rat prostate glands at adulthood. The Prostate 70:689-700. https://doi.org/10.1002/pros.21101
  4. Asadi N, Bahmani M, Kheradmand A, Rafieian-Kopaei M. 2017. The impact of oxidative stress on testicular function and the role of antioxidants in improving it: A review. Journal of Clinical and Diagnostic Research 11:IE01-IE05. https://doi.org/10.7860/JCDR/2017/23927.9886
  5. Axelstad M, Boberg J, Nellemann C, Kiersgaard M, Jacobsen PR, Christiansen S, Hougaard KS, Hass U. 2011. Exposure to the widely used fungicide mancozeb causes thyroid hormone disruption in rat dams but no behavioral effects in the offspring. Toxicological Sciences 120:439-446. https://doi.org/10.1093/toxsci/kfr006
  6. Ayala A, Munoz MF, Arguelles S. 2014. Lipid peroxidation: Production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxidative Medicine and Cellular Longevity 2014:360438.
  7. Berger T. 1990. Pisum sativum agglutinin used as an acrosomal stain of porcine and caprine sperm. Theriogenology 33:689-695. https://doi.org/10.1016/0093-691X(90)90546-6
  8. Blommaert D, Sergeant N, Delehedde M, Jouy N, Mitchell V, Franck T, Donnay I, Lejeune JP, Serteyn D. 2019. Expression, localization, and concentration of A-kinase anchor protein 4 (AKAP4) and its precursor (proAKAP4) in equine semen: Promising marker correlated to the total and progressive motility in thawed spermatozoa. Theriogenology 131:52-60. https://doi.org/10.1016/j.theriogenology.2019.03.011
  9. Calogero AE, Condorelli RA, Russo GI, La Vignera S. 2017. Conservative nonhormonal options for the treatment of male infertility: Antibiotics, anti-inflammatory drugs, and antioxidants. Biomedical Research International. 2017:4650182.
  10. Chandrasekhar Y, Holland MK, D'Occhio MJ, Setchell BP. 1985. Spermatogenesis, seminal characteristics and reproductive hormone levels in mature rams with induced hypothyroidism and hyperthyroidism. Journal of Endocrinology 105:39-46. https://doi.org/10.1677/joe.0.1050039
  11. Dearfield KL. 1994. Ethylene thiourea (ETU). A review of the genetic toxicity studies. Mutation Research/Reviews in Genetic Toxicology 317:111-132. https://doi.org/10.1016/0165-1110(94)90020-5
  12. Devi PA, Paramasivam M, Prakasam V. 2015. Degradation pattern and risk assessment of carbendazim and mancozeb in mango fruits. Environmental Monitoring and Assessment 187:1-6. https://doi.org/10.1007/s10661-014-4167-x
  13. Gao Y, Lee WM, Cheng CY. 2014. Thyroid hormone function in the rat testis. Frontiers in Endocrinology 5:188.
  14. Gavrilescu M, Demnerova K, Aamand J, Agathos S, Fava F. 2015. Emerging pollutants in the environment: Present and future challenges in biomonitoring, ecological risks and bioremediation. New Biotechnology 32:147-156. https://doi.org/10.1016/j.nbt.2014.01.001
  15. Greenlee AR, Ellis TM, Berg RL. 2004. Low-dose agrochemicals and lawn-care pesticides induce developmental toxicity in murine preimplantation embryos. Environmental Health Perspectives 112:703-709. https://doi.org/10.1289/ehp.6774
  16. Hernandez A. 2018. Thyroid hormone role and economy in the developing testis. Vitamins and Hormones 106:473-500. https://doi.org/10.1016/bs.vh.2017.06.005
  17. Jacobsen OS, Bossi R. 1997. Degradation of ethylenethiourea (ETU) in oxic and anoxic sandy aquifers. FEMS Microbiology Reviews 20:539-544. https://doi.org/10.1111/j.1574-6976.1997.tb00336.x
  18. Kozlovsky P, Gefen A. 2013. Sperm penetration to the zona pellucida of an oocyte: A computational model incorporating acrosome reaction. Computer Methods Biomechanics and Biomedical Engineering 16:1106-1111. https://doi.org/10.1080/10255842.2013.768618
  19. Ksheerasagar RL, Kaliwal BB. 2003. Temporal effects of mancozeb on testes, accessory reproductive organs and biochemical constituents in albino mice. Environmental Toxicology and Pharmacology 15:9-17. https://doi.org/10.1016/j.etap.2003.08.006
  20. Ksheerasagar RL, Kaliwal BB. 2010. Effect of mancozeb on thyroid, testis, accessory reproductive organs and biochemical constituents in albino mice. Recent Research in Science and Technology 2:7-17.
  21. Lin YN, Roy A, Yan W, Burns KH, Matzuk MM. 2007. Loss of zona pellucida binding proteins in the acrosomal matrix disrupts acrosome biogenesis and sperm morphogenesis. Molecular and Cell Biology 27:6794-6805. https://doi.org/10.1128/MCB.01029-07
  22. Mandic-Rajcevic S, Rubino FM, Colosio C. 2020. Establishing health-based biological exposure limits for pesticides: A proof of principle study using mancozeb. Regulatory Toxicology and Pharmacology 115:104689.
  23. Maran RR, Arunakaran J, Aruldhas MM. 2000. T3 directly stimulates basal and modulates LH induced testosterone and oestradiol production by rat Leydig cells in vitro. Endocrine Journal 47:417-428. https://doi.org/10.1507/endocrj.47.417
  24. Marinovich M, Guizzetti M, Ghilardi F, Viviani B, Corsini E, Galli CL. 1997. Thyroid peroxidase as toxicity target for dithiocarbamates. Archives of Toxicology 71:508-512. https://doi.org/10.1007/s002040050420
  25. Martins CF, Dode MN, Bao SN, Rumpf R. 2007. The use of the acridine orange test and the TUNEL assay to assess the integrity of freeze-dried bovine spermatozoa DNA. Genetics and Molecular Research 6:94-104.
  26. Miki K, Willis WD, Brown PR, Goulding EH, Fulcher KD, Eddy EM. 2002. Targeted disruption of the Akap4 gene causes defects in sperm flagellum and motility. Developmental Biology 248:331-342. https://doi.org/10.1006/dbio.2002.0728
  27. Mingo V, Lotters S, Wagner N. 2017. The impact of land use intensity and associated pesticide applications on fitness and enzymatic activity in reptiles-A field study. Science of the Total Environment 590:114-124. https://doi.org/10.1016/j.scitotenv.2017.02.178
  28. Mohammadi-Sardoo M, Mandegary A, Nabiuni M, Nematollahi-Mahani SN, Amirheidari B. 2018. Mancozeb induces testicular dysfunction through oxidative stress and apoptosis: Protective role of N-acetylcysteine antioxidant. Toxicology and Industrial Health 34:798-811. https://doi.org/10.1177/0748233718778397
  29. Pandey SP, Mohanty B. 2015. The neonicotinoid pesticide imidacloprid and the dithiocarbamate fungicide mancozeb disrupt the pituitary-thyroid axis of a wildlife bird. Chemosphere 122:227-234. https://doi.org/10.1016/j.chemosphere.2014.11.061
  30. Paramasivam M, Chandrasekaran S. 2013. Dynamics and residues of mixed formulation of fenamidone and mancozeb in gherkin field ecosystem. Ecotoxicology and Environmental Safety 98:292-296. https://doi.org/10.1016/j.ecoenv.2013.08.006
  31. Pirozzi AVA, Stellavato A, La Gatta A, Lamberti M, Schiraldi C. 2016. Mancozeb, a fungicide routinely used in agriculture, worsens nonalcoholic fatty liver disease in the human HepG2 cell model. Toxicology Letters 249:1-4. https://doi.org/10.1016/j.toxlet.2016.03.004
  32. Pursel VG, Johnson LA. 1976. Frozen boar spermatozoa: Methods of thawing pellets. Journal of Animal Science 42: 927-931. https://doi.org/10.2527/jas1976.424927x
  33. Rey F, Gonzalez M, Zayas MA, Stoker C, Durando M, Luque EH, Munoz-de-Toro M. 2009. Prenatal exposure to pesticides disrupts testicular histoarchitecture and alters testosterone levels in male Caiman latirostris. General and Comparative Endocrinology 162:286-292. https://doi.org/10.1016/j.ygcen.2009.03.032
  34. Rijntjes E, Swarts HJ, Anand-Ivell R, Teerds KJ. 2009. Prenatal induced chronic dietary hypothyroidism delays but does not block adult-type Leydig cell development. American Journal of Physiology-Endocrinology and Metabolism 296:305-314. https://doi.org/10.1152/ajpendo.90750.2008
  35. Rossi G, Palmerini MG, Macchiarelli G, Buccione R, Cecconi S. 2006. Mancozeb adversely affects meiotic spindle organization and fertilization in mouse oocytes. Reproductive Toxicology 22:51-55. https://doi.org/10.1016/j.reprotox.2005.11.005
  36. Runkle J, Flocks J, Economos J, Dunlop AL. 2017. A systematic review of mancozeb as a reproductive and developmental hazard. Environment International 99:29-42. https://doi.org/10.1016/j.envint.2016.11.006
  37. Spinaci M, Nerozzi C, Mislei B, Blanco-Prieto O, Mari G, Galeati G, Bucci D. 2022. Impact of glyphosate and its formulation Roundup® on stallion spermatozoa. Theriogenology 79:197-203. https://doi.org/10.1016/j.theriogenology.2021.12.003
  38. Srivastava AK, Ali W, Singh R, Bhui K, Tyagi S, Al-Khedhairy AA, Srivastava PK, Musarrat J, Shukla Y. 2012. Mancozeb-induced genotoxicity and apoptosis in cultured human lymphocytes. Life Sciences 90:815-824. https://doi.org/10.1016/j.lfs.2011.12.013
  39. Suarez ERL, Geissen V, Sanchez AJ, Chan RAC, Bello-Mendoza R. 2013. Formation and decay of ethylenethiourea (ETU) in soil and water under tropical conditions. Journal of Plant Nutrition and Soil Science 176:40-46. https://doi.org/10.1002/jpln.201200014
  40. Veeramachaneni DR. 2000. Deteriorating trends in male reproduction: Idiopathic or environmental? Animal Reproduction Science 60:121-130. https://doi.org/10.1016/S0378-4320(00)00113-5
  41. Wagner MS, Wajner SM, Maia AL. 2009. Is there a role for thyroid hormone on spermatogenesis? Microscopy Research and Technique 72:796-808. https://doi.org/10.1002/jemt.20759
  42. Wang Z, Kottawatta KS, Kodithuwakku SP, Fernando TS, Lee YL, Ng EH, Yeung WS, Lee KF. 2021. The fungicide Mancozeb reduces spheroid attachment onto endometrial epithelial cells through downregulation of estrogen receptor β and integrin β3 in Ishikawa cells. Ecotoxicology and Environmental Safety 208:111606.
  43. WHO (World Health Organization). 1988. Dithiocarbamate pesticides, ethylenethiourea and propylenethiourea: A general introduction. Environmental Health Criteria No 78. WHO, Geneva, Swiss.
  44. Xu K, Yang L, Zhang L, Qi H. 2020. Lack of AKAP3 disrupts integrity of the subcellular structure and proteome of mouse sperm and causes male sterility. Development 147:dev181057.
  45. Yi YJ, Adikari AADI, Moon ST, Lee SM, Heo JM. 2021. Effects of lipopolysaccharides on the maturation of pig oocytes. Korean Journal of Agricultural Science 48:163-170. https://doi.org/10.7744/KJOAS.20210010
  46. Zhao W, Li Z, Ping P, Wang G, Yuan X, Sun F. 2018. Outer dense fibers stabilize the axoneme to maintain sperm motility. Journal of Cellular and Molecular Medicine 22:1755-1768. https://doi.org/10.1111/jcmm.13457
  47. Zhou R, Wu J, Liu B, Jiang Y, Chen W, Li J, He Q, He Z. 2019. The roles and mechanisms of Leydig cells and myoid cells in regulating spermatogenesis. Cellular and Molecular Life Sciences 76:2681-2695. https://doi.org/10.1007/s00018-019-03101-9