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A. Faridha ¹, K. Faisal ¹, M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, Bharathidasan University, Tiruchirappalli - 620 024, IN

Abstract

In the background of the decreasing trend of male reproductive health in men and animals, the present study was aimed at finding gravimetric, histopathologoical and histometric changes in the testis of Swiss mouse in response to treatment of aflatoxin B₁ (AFB₁) in a chronic toxicity testing over different periods of time and at finding recovery, if any, after cessation of the treatment. AFB₁ suspended in corn oil and ethanol, was administered through intraperitoneal route to 90 day old Swiss mouse at a daily dose of 50mg/kg body weight for 7, 15, 35, 45 days. In the recovery group the mice were kept, after cessation of the treatment, under observation for 35 or 70 days. The testicles and seminal vesicles of the animals were subjected to histopathological analysis adopting paraffin/resin embedding and light microscopy. Computer assisted histometric analysis of several parameters was also made. In general there was little impact of the treatment in the animals treated for 7 days. In the animals treated for 15, 35 and 45 days, there was duration-dependent regression of the testis and seminal vesicles. Histopathological changes were observed in both the spermatogenic and androgenic compartments of the testis. Fragmentation of chromatin of pachytene spermatocytes, generation of uni- and multinucleate gaint cells, and premature loss of spermatids and spermatocytes were observed. Whereas the seminiferous tubules regressed, the Leydig cells underwent hypertrophy and distortion of shape of the nucleus. Generation of multiple micronucleate gaint cells and extensive loss of germ cells from the seminiferous epithelium were observed. Fertility of the mice was severely hampered. The various changes are discussed in relation to the possible effects of AFB₁ on the microtubules of the spindle fibers and the DNA of chromatin. Since spermatogonia and Sertoli cells are not vulnerable targets to AF toxicity, the treated animals recovered over a period of time.

Keywords

Aflatoxin B1, Male Reproductive Toxicity, Testis, Seminal Vesicles.

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K. Faisal ¹, S. Parveen ², R. Rajendran ³, R. Girija ¹, V. S. Periasamy ¹, B. Kadalmani ¹, A. Puratchikody ⁴, K. Ruckmani ⁴, B. M. J. Pereira ², M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620 024, IN
2 Department of Bioscience and Biotechnology, Reproductive Biology Laboratory, Indian Institute of Technology Roorke 247 667, IN
3 Green Chem, Attibele Industrial Area, Bangalore - 562 107, IN
4 Department of Pharmaceutical Engineering & Technology Bharathidasan Institute of Technology, Tiruchirappalli - 620 024, IN

Abstract

Quassia amara is a medicinal plant with several pharmacological properties. More recently, it has come to be known in the context of its potential biopesticidal application and the antimalarial property in two of its secondary metabolites, quassin and neo-quassin. There are also preliminary reports implicating this plant in male reproductive toxic effects, to the extent that the plant could as well be tested for male contraceptive efficacy. Therfore, we subjected the methanolic extract of the bark of this tree for male reproductive toxicological evaluation. This paper reports several derangements in the Cauda epididymidal sperm including a hither - to unreported one.

Keywords

Neo-Quassin, Quassia amara, Quassin, Sperm Abnormalities.

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K. Faisal , R. Girija , M. A. Akbarsha

Abstract

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.

Keywords

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

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K. Faisal ¹, M. A. Akbarsha ¹

Affiliations
1 Department of Animal Science, School of Life Sciences, Bharathidasan University, Tiruchirappalli - 620024, Tamil Nadu, IN

Abstract

Ubiquitination is believed to play a critical role in removal of dead and/or defective spermatozoa in normal and, more importantly, under circumstances when such spermatozoa are produced in large numbers due to genetic defects or toxic manifestations. Ubiquitination under such instances would involve specific gene expressions, many of which are not yet clearly understood. In an exhaustive study in Swiss mouse model to find the spermatotoxic effect of quassin, a diterpene compound isolated from Quassia amara, we found most of the spermatozoa to be abnormal in morphology and unviable. In the present study, we aimed at analysing the transcriptional profile of three selected genes, Ubb, Ube2c and Psmb8, involved in the ubiquitin proteolytic pathway in the testis and epididymal segments of Q. amara bark methanol extract treated mice adopting semi-quantitative RT-PCR and to study the level of DNA damage of the treated mouse spermatozoa. The results revealed that the treatment induced considerable damage to the sperm DNA. All the three genes studied showed marked increase in their levels of expression in the treated mice compared to the corresponding controls. Thus, this study shows that Q. amara methanol extract is causative of sperm DNA damage and defective spermatozoa and, in such cases, the expression of specific genes concerned with ubiquitination pathway is increased, implying that ubiquitination-proteosomal degradation is involved in the processing of dead/defective spermatozoa.

Keywords

Defective Sperm, Proteosomal Degradation, Quassin, Quassia amara, Spermatotoxicity, Ubiquitination.

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