Sperm characteristics and ultrastructure of testes of rats after long-term treatment with the methanol subfraction of Carica papaya seeds

The contraceptive efficacy of Carica papaya seeds after short-term evaluation has been well established. We have examined the safety and mechanism of contraception in rats after long-term treatment with the methanol subfraction （MSF） of C. papaya seeds. The test substance was administered orally to the male albino rats （n = 40） at 50 mg per kg body weight each day for 360 days. Control animals （n = 40） received olive oil as a vehicle. Recovery was assessed up to 120 days after treatment withdrawal. Sperm parameters, serum testosterone levels, fertility, histology and ultrastructure of the testis, haematology and serum clinical chemistry were evaluated to establish the safety and efficacy of the test substance. Safety of long-term treatment was evidenced by unaltered health status, organ weight, haematology and clinical chemistry, and by an increase in body weight. The mechanism of contraception was shown by reduction in nuclear and cytoplasmic volume, normal nuclear characteristics and vacuolization in the cytoplasmic organelles of the Sertoli cells, as well as nuclear degeneration in spermatocytes and spermatids indicating disturbed spermatogenesis. Leydig cells were normal. Initial effects were observed in Sertoli cells at 60 days of treatment. Spermatocytes and spermatids were affected after 120-240 days of treatment. A significant decline in sperm count and viability, total inhibition of sperm motility, increased numbers of sperm abnormalities, normal serum testosterone levels and 100% sterility were evident after 60 days of treatment. All the altered parameters, including percent fertility, were restored to control level 120 days after treatment withdrawal. It is concluded that the MSF is safe for long-term treatment and the mechanism of contraception is shown by its effect on spermatid differentiation in the testis, possibly mediated by the Sertoli cell factors.

Sperm motility inhibitory effect of the benzene chromatographic fraction of the chloroform extract of the seeds of Carica papaya in langur monkey, Presbytis entellus entellus

Aim： To assess the contraceptive efficacy of the benzene chromatographic fraction of the chloroform extract of the seeds of Carica papaya in langur monkeys. Methods： The test substance was given p.o. to five monkeys at 50 mg/kg body weight/day for 360 days. Control animals （n = 3） received olive oil as vehicle. Sperm parameters as per World Health Organization standards, sperm functional tests, morphology of testis and epididymis, haematology, clinical biochemistry, serum testosterone and libido were evaluated. Following completion of 360 days treatment the animals were withdrawn from the treatment and the recovery pattern was assessed by semen analysis and sperm functional tests. Results： Total inhibition of sperm motility was observed following 60 days of treatment that continued until 360 days study period. Sperm count, percent viability and percent normal spermatozoa showed a drastic decline following 30 days of treatment. Sperm morphology showed predominant mid piece abnormalities. Sperm functional tests scored in sterile range. Histology and ultrastructure of testis revealed vacuolization in the Sertoli cells and germ cells. Loss of cytoplasmic organelles was evident in spermatocytes and round spermatids. Histology and ultrastructure of epididymis of treated animals were comparable to those of control animals. Hematological and serum clinical parameters and testosterone levels fluctuated within the control range throughout the study period. Recovery was evident following 60--120 days of treatment withdrawal. Conclusion- The results suggest that the benzene chro- matographic fraction of the chloroform extract of the seeds of Carica papaya shows contraceptive efficacy without adverse toxicity, mediated through inhibition of sperm motility. （Asian JAndro12008 Mar; 10： 298-306）

Effects of Coated Capillary Column, Derivatization, and Temperature Programming on the Identification of Carica papaya Seed Extract Composition Using GC/MS Analysis

Some of the common practices to assess the composition of plant extract,including Carica papaya seed extract(CPSE)are direct injection of the extract,compound separation using polyethylene glycol capillary column(DB-WAX),and non-linear-temperature programming(NLTP)of GC/MS analysis.This study specifically compared the coating of capillary column,sample derivatization,and temperature programming of GC/MS to determine the composition of CPSE.The retention indices(RI)of the detected compounds were determined and compared to the reference RI.In particular,5%phenyl-95%methylpolysiloxane(HP-5MS)-,DB-WAX-,and biscyanopropyl polysiloxane(HP-88)-coated capillary columns were used to identify the composition of CPSE.For this study,HP-5MS column,which separated the highest number of compounds(26 compounds)from CPSE,was deemed as the most suitable column.The GC/MS analysis of derivatized CPSE identified 21 compound groups,where fatty acids and fatty acid methyl esters served as the major compounds(80.23%),followed by these compounds in decreasing order:amides>nitriles>sterols>fatty aldehydes>organic acids.A stronger correlation determination between the carbon number and alkane retention time of linear-temperature programming(LTP)(R^(2)=0.9859)was found,as compared to its correlation determination with NLTP(R^(2)=0.9175),which exhibited an almost equal RI of LTP to the reference RI.Conclusively,GC/MS analysis for the derivatized CPSE using HP-5MS column separation and LTP is highly recommended.