Aspects of Male Reproductive Toxic Effects of Quassia amara L.: Histopathological and Ultrastructural Study in Mouse


  • Bharathidasan University, Department of Animal Science, Tiruchirappalli, Tamil Nadu, 620 024, India
  • Mercy College, Palakkad, Kerala, 678006, India


Quassia amara is a Brazilian medicinal plant traditionally used as an herbal remedy for a variety of diseases. Quassin is the major secondary chemical compound in it, which is under investigation as an anti-malarial compound. There are preliminary studies implicating Q. amara in male reproductive toxicity. Hence, this elaborate study was undertaken to find the male reproductive toxic effects of Q. amara and quassin. Methanolic extract of Q. amara bark and quassin were administered to male mice for 35 days. The cauda epididymidal sperm, testis and epididymis were subjected to analysis adopting light and transmission electron microscopy. Sperm counts, motility and viability decreased. The incidence of sperm with abnormal morphologies increased. There was premature loss of germ cells from the seminiferous tubules, which correlated with Sertoli cell pathology. The Leydig cells were vacuolated, suggesting a hypo-androgen status. The epididymis indicated severe histopathological changes, including reflection of a mitogenic / tumorigenic effect of Q. amara. Thus, this study indicates that Q. amara, when applied as a therapeutic, could be severely toxic to male reproductive mechanisms and the principal mechanism of action appears to be through Leydig cells. Quassin, the major secondary metabolite responsible for the therapeutic potential of Q. amara, is also the one causative of the male reproductive toxic effects. Thus, this study substantiates the claim that Q. amara is toxic to male reproductive system.


Epididymis, Histopathology, Sperm, Testis, Quassia amara, Quassin

Subject Discipline

Obstetrics and Gynaecology

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Abe K, Takano H. (1989) Early degeneration of the epithelial cells in the initial segment of the epididymal duct in mice after efferent duct cutting. Arch Histol Cytol. 52: 299–310.

Abe K, Takano H, Ito T. (1982) Appearance of peculiar epithelial cells in the epididymal duct of the mouse ligated epididymis. Biol Reprod. 26: 501–509.

Adamali HI, Hermo L. (1996) Apical and narrow cells are distinct cell types differing in their structure, distribution, and functions in the adult rat epididymis. J Androl. 17: 208–222.

Adamali HI, Somani IH, Huang JQ, Mahuran D, Gravel RA, Trasler JM, Hermo L. (1999a) Abnormalities in cells of the testis, efferent ducts, and epididymis in juvenile and adult mice with beta-hexosaminidase A and B deficiency. J Androl. 20: 779–802.

Adamali HI, Somani IH, Huang JQ, Gravel RA, Trasler JM, Hermo L. (1999b) Characterization and development of the regional- and cellular-specific abnormalities in the epididymis of mice with beta-hexosaminidase A deficiency. J Androl. 20: 803–824.

Agnes VF, Akbarsha MA. (2001) Pale vacuolated epithelial cells in the epididymis of aflatoxin-treated mice. Reproduction 122: 629–641.

Ajaiyeoba EO, Krebs HC. (2003) Antibacterial and antifungal activities of Quassia undulata and Quassia amara extracts in vitro. African J Medicine and Medical Sciences 32: 353–356.

Ajaiyeoba EO, Abalogu UI, Krebs HC, Oduola AM. (1999) In vivo antimalarial activities of Quassia amara and Quassia undulata plant extracts in mice. J Ethnopharmacol. 67: 321–325.

Akbarsha MA, Murugaian P. (2000) Aspects of the male reproductive toxicity/male antifertility property of andrographolide in albino rat: effect on testis and cauda epididymidal spermatozoa. Phytother Res. 14: 1–4.

Akbarsha MA, Stanley A, Averal HI. (1995) Effect of vincristine on Leydig cells and accessory reproductive organs. Curr Sci. 68: 1053–1056.

Akbarsha MA, Palanisamy M, Murugaian P, Lakshmi Latha PN. (1998) Ursolic acid generates symplasts in rat spermatogenic clones. Phytother Res. 12: 32–36.

Akbarsha MA, Latha PN, Murugaian P. (2000) Retention of cytoplasmic droplet by rat cauda epididymal spermatozoa after treatment with cytotoxic and xenobiotic agents. J Reprod Fertil. 120: 385–390.

Akbarsha MA, Faisal K, Radha A (2015) The epididymis: Structure and function. In: Singh SK editor. Mammalian Endocrinology and Male Reproductive Biology. CRC Press/Taylor & Francis. pp 115-165.

Apers S, Cimanga K, Vanden Berghe D, Van Meenen E, Longanga AO, Foriers A, Vlietinck A, Pieters L. (2002) Antiviral activity of simalikalactone D, a quassinoid from Quassia africana. Planta Med. 68: 20–24.

Aruldhas MM, Subramanian S, Sekhar P, Vengatesh G, Govindarajulu P, Akbarsha MA. (2006) In vivo spermatotoxic effect of chromium as reflected in the epididymal epithelial principal cells, basal cells, and intraepithelial macrophages of a nonhuman primate (Macaca radiata Geoffroy). Fertil Steril. 86: 1097–1105.

Bertani S, Houel E, Bourdy G, Stien D, Jullian V, Landau I, Deharo E. (2007) Quassia amara L. (Simaroubaceae) leaf tea: effect of the growing stage and desiccation status on the antimalarial activity of a traditional preparation. J Ethnopharmacol. 111: 40–42.

Boekelheide K. (2005) Mechanisms of toxic damage to spermatogenesis. JNCI Monographs. 34: 6–8.

Bremner WJ, Millar MR, Sharpe RM, Saunders PT. (1994) Immunohistochemical localization of androgen receptors in the rat testis: evidence for stage-dependent expression and regulation by androgens. Endocrinology 135: 1227–1234.

Bronson FH, Dagg CP, Snell GD. (1975) Reproduction. In: Green EL (eds), Biology of the Laboratory Mouse. New York: Dover Publications Inc. pp 187–204.

Capasso R, Izzo AA, Pinto L, Bifulco T, Vitobello C, Mascolo N. (2000) Phytotherapy and quality of herbal medicines. Fitoterapia 71: S58–S65.

Carlsen E, Giwercma, A, Keiding S, Skakkebaek NE. (1992) Evidence for decreasing quality of semen during the past 50 years. BMJ (Clinical research ed.) 305: 609–613.

Carlsen E, Giwercman A, Skakkebaek NE, Keiding N. (1993) Decreasing quality of semen. BMJ (Clinical research ed.) 306: 461.

Clermont Y. (1961) Incorporation of thymidine H3 in epithelial cells of the ductus epididymis in albino rats. Anat Rec. 139: 2A (Abstr).

Clermont Y, Flannery J. (1970) Mitotic activity in the epithelium of the epididymis in young and old adult rats. Biol Reprod. 3: 283–292.

Cooper TG. (1998) Interactions between epididymal secretions and spermatozoa. J Reprod Fertil. Suppl. 53: 119–136.

De Gendt K, Swinnen JV, Saunders PT, Schoonjans L, Dewerchin M, Devos A, Tan K, Atanassova N, Claessens F, Lécureuil C, Heyns W, Carmeliet P, Guillou F, Sharpe RM, Verhoeven G. (2004) A Sertoli cell-selective knockout of the androgen receptor causes spermatogenic arrest in meiosis. Proc Natl Acad Sci USA 101: 1327–1332.c

Diakanamwa C, Diallo D, Vanhaelen-Fastre R, Vanhaelen M, Ottinger R. (1993) 14-Hydroxychapparinone, a new quassinoid from Hannoa chlorantha. J Nat Prod. 56: 1817–1820.

Faisal K, Parveen S, Rajendran R, Girija R, Periasamy VS, Kadalmani B, Puratchikody A, Ruckmani K, Pereira BMJ, Akbarsha MA. (2006) Male reproductive toxic effect of Quassia amara: Observations on mouse sperm. J Endocrinol Reprod. 10: 66–69.

Flores G, Hilje L, Mora GA, Carballo M. (2008) Antifeedant activity of botanical crude extracts and their fractions on Bemisia tabaci (Homoptera: Aleyrodidae) adults: III. Quassia amara (Simaroubaceae). Revista de Biología Tropical. 56: 2131–2146.

Goyal HO, Hutto V, Maloney MA. (1994) Effects of androgen deprivation in the goat epididymis. Acta Anat. 150: 127–135.

Grieco PA, Parker DT, Nargund RP. (1988) Total synthesis of a highly oxygenated quassinoid, (.+-.)-klaineanone. J Am Chem Soc. 110: 5568–5569.

Handelsman DJ. (2001) Estrogens and falling sperm counts. Reprod Fertil Dev. 13: 317–324.

Hayat MA. (1981) The production of artifacts. Ultrastructural Pathol. 2: 93.

Hermo L, Dworkin J, Oko R. (1988) Role of epithelial clear cells of the rat epididymis in the disposal of the contents of cytoplasmic droplets detached from spermatozoa. Am J Anat. 183: 107–124.

Hermo L, Adamali HI, Mahuran D, Gravel RA,Trasler JM. (1997) Beta-Hexosaminidase immunolocalization and alpha- and beta-subunit gene expression in the rat testis and epididymis. Mol Reprod Dev. 46: 227–242.

Hess RA, Thurston RJ. (1977) Ultrastructure of epithelial cells in the epididymal region of the Turkey (Meleagris gallopavo). J Anat. 124: 765–778.

Husain GM, Singh PN, Singh RK, Kumar V. (2011) Antidiabetic activity of standardized extract of Quassia amara in nicotinamide–streptozotocin-induced diabetic rats. Phytother Res. 25: 1806–1812.

Itoh M, Miyamoto K, Satriotomo I, Takeuchi Y. (1999) Spermatic granulomata are experimentally induced in epididymides of mice receiving high-dose testosterone implants. I. A light-microscopical study. J Androl. 20: 551–558.

Jackson AE, O’Leary PC, Ayers MM, de Kretser DM. (1986) The effects of ethylene dimethane sulphonate (EDS) on rat Leydig cells: evidence to support a connective tissue origin of Leydig cells. Biol Reprod. 35: 425–437.

Karnovsky MJ. (1965) A formaldehyde-glutaraldehyde fixative of high osmolarity for use in electronmicroscopy. J Cell Biol. 25: 137A.

Kupchan SM, Streelman DR. (1976) Quassimarin, a new antileukemic quassinoid from Quassia amara. J Org Chem. 41: 3481–3482.

Martan J, Risley PL. (1963) The epididymis of mated and unmated rats. J Morphol. 113: 1–15.

Meistrich ML, Goldstein LS, Wyrobek AJ. (1985) Long-term infertility and dominant lethal mutations in male mice treated with adriamycin. Mutat Res. 152: 53–65.

Mortimer D, Mortimer ST. (2005) Laboratory investigation of the infertile male. In: Brinsden PR editor. Textbook of in Vitro Fertilization and Assisted Reproduction, 3rd edn. London and New York: CRC Press, pp 61–91.

Nakai M, Hess RA. (1994) Morphological changes in the rat Sertoli cell induced by the microtubule poison carbendazim. Tissue Cell. 6: 917–927.

Nau JY. (2016) French bio-pyrates and Quassia amara preparation against malaria. Rev Med Suisse. 17: 370–371.

Negi AS, Kumar JK, Luqman S, Shanker K, Gupta MM, Khanuja SP. (2008) Recent advances in plant hepatoprotectives: a chemical and biological profile of some important leads. Med Res Rev. 28: 746–772.

Njar VC, Alao TO, Okogun JI, Raji Y, Bolarinwa AF, Nduka EU. (1995) Antifertility activity of Quassia amara: quassin inhibits the steroidogenesis in rat Leydig cells in vitro. Planta Med. 61: 180–182.

Oakberg EF. (1956) Duration of spermatogenesis in the mouse and timing of stages of the cycle of the seminiferous epithelium. Am J Anat. 99: 507–516.

Park MH, Maeda M, Komura H, Nakanishi K, Nomoto K. (1987) Acute insecticidal activity of quassin and its congeners against the American cockroach. Chem Pharm Bull. 35: 3082–3085.

Parveen S, Das S, Kundra CP, Pereira BMJ. (2003) A comprehensive evaluation of the reproductive toxicity of Quassia amara in male rats. Reprod Toxicol. 17: 45–50.

Raji Y, Bolarinwa AF. (1997) Antifertility activity of Quassia amara in male rats - in vivo study. Life Sci. 61: 1067–1074.

Richburg JH. (2000) The relevance of spontaneous- and chemically-induced alterations in testicular germ cell apoptosis to toxicology. Toxicol Lett. 15: 79–86.

Robaire B, Fan X. (1998) Regulation of apoptotic cell death in the rat epididymis. J Reprod Fertil Suppl. 53: 211–214.

Robaire B, Hermo L. (1988) Efferent ducts, epididymis, and vas deferens: structure, functions and their regulation. In: Knobil E, Neill JD (eds) The Physiology of Reproduction. Raven Press, New York, pp 999–1080.

Robaire B, Viger RS. (1995) Regulation of epididymal epithelial cell functions. Biol Reprod. 52: 226–236.

Sharpe RM, Skakkebaek NE. (1993) Are oestrogens involved in falling sperm counts and disorders of the male reproductive tract? Lancet. 341: 1392–1395.

Skakkebaek NE, Rajpert-De Meyts E, Jorgensen N, Carlsen E, Petersen PM, Giwercman A, Andersen AG, Jensen

TK, Andersson AM, Muller J. (1998) Germ cell cancer and disorders of spermatogenesis: an environmental connection? APMIS. 3: 11–12.

Slama R, Jegou B, Cordier S. (2006) Breakthroughs in the study of the influence of environmental factors on male reproductive health. Revue d’epidemiologie et de sante publique 54: 167–174.

Spector DL, Goldman RD, Leinwand LA. (1997) Cells: A Laboratory Manual. Volume 1: Culture and Biochemical Analysis of Cells. USA: Cold Spring Harbor Laboratory Press.

Sun EL, Flickinger CJ. (1979) Development of cell types and of regional differences in the postnatal rat epididymis. Am J Anat. 154: 27–55.

Toma W, Gracioso Jde S, de Andrade FD, Hiruma-Lima CA, Vilegas W, Souza Brito AR. (2002) Antiulcerogenic activity of four extracts obtained from the bark wood of Quassia amara L. (Simaroubaceae). Biol Pharm Bull. 25: 1151–1155.

Toshimori K, Araki S, Oura C. (1990) Epithelial cells with vacuoles containing 54,000 dalton sialoglycoprotein in the mouse epididymal duct. Arch Histol Cytol. 53: 333–338.

Trager W, Polonsky J. (1981) Antimalarial activity of quassinoids against chloroquine-resistant Plasmodium falciparum in vitro. Am J Tropical Med Hyg. 30: 531–537.

Trasler JM, Hermo L, Robaire B. (1988) Morphological changes in the testis and epididymis of rats treated with cyclophosphamide: a quantitative approach. Biol Reprod. 38: 463–479.

Turner TT. (1995) On the epididymis and its role in the development of the fertile ejaculate. J Androl. 16: 292–298.

van Valkenburg JLCH, Bunyapraphatsara N. (2002) Plant Resources of South-East Asia No.12(2). Medicinal and Poisonous Plants 2. Indonesia: Prosea Foundation.

Veeramachaneni DN. (2000) Deteriorating trends in male reproduction: idiopathic or environmental? Anim Reprod Sci. 60–61: 121–130.

Veri JP, Hermo L, Robaire B. (1994) Immunocytochemical localization of glutathione S-transferase Yo subunit in the rat testis and epididymis. J Androl 15: 415–434.

Vigneron M, Deparis X, Deharo E, Bourdy G. (2005) Antimalarial remedies in French Guiana: a knowledge attitudes and practices study. J Ethnopharmacol. 98: 351–360.

Wang JS, Groopman JD. (1999) DNA damage by mycotoxins. Mutat Res. 424: 167–181.

Winters SJ, Wang C, Abdelrahaman E, Hadeed V, Dyky MA, Brufsky A. (2006) Inhibin-B levels in healthy young adult men and prepubertal boys: is obesity the cause for the contemporary decline in sperm count because of fewer Sertoli cells? J Androl. 27: 560–564.


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