EFFECT OF INCORPORATING 10% AND 20% MUCUNA ROSTRATA SEED MEAL ON THE HISTOPATHOLOGY OF TESTES AND SOME SPERM CHARACTERISTICS OF ALBINO WISTAR RATS.

International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 378 EFFECT OF INCORPORATING 10% AND 20% MUCUNA ROSTRATA SEED MEAL ON THE HISTOPATHOLOGY OF TESTES AND SOME SPERM CHARACTERISTICS OF ALBINO WISTAR RATS. EDET, EMMANUEL A.1, AKPET, IMAOBONG N.2, BOBSON, PROMISEGODSFAVOUR M.3 1. Department of Science Technology, Akwa Ibom State Polytechnic, Ikot Osurua, Akwa Ibom State, Nigeria, 2347084847561, [email protected] 2. Department of Chemical Sciences, Akwa Ibom State Polytechnic, Ikot Osurua, Akwa Ibom State, Nigeria, 2348023222111, [email protected] 3. Department of Chemical Sciences, Akwa Ibom State Polytechnic, Ikot Osurua, Akwa Ibom State, Nigeria, 2347035746429, [email protected] ARTICLE INFO ABSTRACT ©2025 RS Publication Paper ID: IJASTR68E8E16404190 Received: 2025-09-15 Published: 2025-10-15 DOI: https://dx.doi.org/ 10.5281/zenodo.1736 8717 Page No: 378-386 The effect of incorporating 10% and 20% roasted Mucuna rostrata seed meal for 28 days on the histopathology of the testes and some sperm characteristics of albino Wistar rats was evaluated. Sixty (60) sexually experienced male albino rats of body weight between 136g and 139g were randomly selected into three study groups; A (Control), B (test Group 1) and C (test Group II). Group A rats were fed with 100% rat mash, Group B and C rats were fed with mash which was incorporated with 10% and 20% roasted Mucuna rostrata seed meal. At the end of 28 days feeding period, the rats were sacrificed. The results showed the sample had no effect on the sperm colour. Viscosity and turbidity remained “normal” for the control and test groups. There were non significant decrease on semen pH, motility, sperm count and morphology of the test groups compared to the control. Histological examination of the testes at 10% and 20% supplementation showed that within the testes, there was progressive increase in both size and number of the Leydig cells as attested to by the density and hyperchromasia of cells within the semniferous tubules, epithelial and interstitial cells of the Leydig. Keywords: Mucuna rostrata, testes, histopatology, sperm characteristics International Journal of Advanced Scientific and Technical Research Available online on http://www.rspublication.com/ijst/index.html ISSN 2249-9954 Cite This Paper: EDET, EMMANUEL A., AKPET, IMAOBONG N. and BOBSON, PROMISEGODSFAVOUR M(2025). "EFFECT OF INCORPORATING 10% AND 20% MUCUNA ROSTRATA SEED MEAL ON THE HISTOPATHOLOGY OF TESTES AND SOME SPERM CHARACTERISTICS OF ALBINO WISTAR RATS.". INTERNATIONAL JOURNAL OF ADVANCED SCIENTIFIC AND TECHNICAL RESEARCH (IJASTR), vol. 15, no. 5, 2025, pp. 378-386 DOI: https://dx.doi.org/10.5281/zenodo.17368717 International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 379 INTRODUCTION: The legume family have been extensively investigated because of the numerous composition most of which have nutritional and medicinal properties. One of such legumes is the Mucuna; commonly called the velvet beans. This legume is an annual or perennial herbaceous, vigorous climbing vine. It can reach a length of 18m if it twine on support or 3-5m if it run on the ground. The velvet beans has fleshly well nodulated roots, long slender and sparsely pubescent stems, numerous alternate and trifoliate hairy leaves and white or greenish-yellow flowers that hang on long clusters. The pods are 9-14cm long. They are hard, curved, slightly ridged and covered with soft velvet hair capable of causing intense skin irritation if dislodged. At maturity each pod produces 3-4 hard marble like seeds which are brown or black in colour. A readily available species of Mucuna is the Mucuna rostrata Consumption of this species by different tribal and ethnic groups in Africa especially in Nigeria is a total reflection of the eating pattern of such communities. Some tribes use the pods and leaves of Mucuna rostrata as vegetables while others use the mature seeds as condiment and main dish. The Efiks and Igbos in the South-South and South Eastern regions of Nigeria roast the mature seeds for 5-10 minutes before grinding and using the flour as recipe for stew, sauce, snacks, porridge, moi-moi, fried cake and soup thickening agent. (Onweluzo and Eilitta (2003) The seeds of Mucuna have been reported to possess inhibitory factors (Siddhuraju et al, 2000) but this does not exclude the seeds from being used as raw materials in agriculture, pharmacy, food, beverage and cosmetics; especially as it has a long history as anti-cancerous agent, worm expellant and aphrodisiac. Despite these potentials, controversies and beliefs still exist among rural populations of Nigeria notably Ibibios and Efiks that consumption of Mucuna rostrata preparations contributes to impotence and infertility in males. International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 380 It is in the light of these controversies that evaluation of the effect of Mucuna rostrata seed meal in male albino rats is conducted with special emphasis on the histology of testes and seminal profile. The findings will undoubtedly lay to rest controversies that discourage the use of Mucuna rostrata as component of food and feed stock MATERIALS AND METHODS Experimental Animals and Animal Grouping A total of sixty (60) healthy, docile and sexually experienced adult male albino rats of the wistar strain were used for the animal experiment. Aged 10 – 12 weeks old and of 136g – 139g body weight, the animals were kept under standard conditions of temperature; 28-31 0 C, photo period; 12 hours natural light/12 hours dark for 24 days. They were divided into 3 study groups of 20 rats each. Where Group A, Group B and Group C were control group, test group 1 and test group 2 respectively. All the animals were marked differently, weighed and confined in 3 different metabolic cages labeled A, B, C. The cages 160cm long, 60cm wide and 30cm high were made of transparent Perspex material with grid bottom and stainless steel top; it had enough space for free movement of the animals, perforation for adequate ventilation and light as well as sieves for separation of feaces and urine Histopathological Examination: The procedure described by Krause (2001) was used to investigate into the structures of the testes of rats in the control and test groups. It involved fixation of the testes in 10% buffered formaldehyde, dehydrating through ascending grades of ethanol, cleaning in xylene, embedding in molten paraffin wax (m.p 56 CG ) and allowing it to solify. The next steps were staining the dried embedded blocks with halmatoxylin and eosin, permanent mounting on International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 381 balsam on the covering-mounting medium and observation with the Leitz, DIALUX research microscope at X200. Collection of sperm cells and determination of seminal parameters: Epididymis samples were removed from the freezer and allowed to thaw slowly in a refrigerator. The samples were then cut open carefully using sterile blades and placed in 0.1M phosphate buffer of pH 7.4. After vigorous shaking for homogeneity and dispersal of sperm cells, sperm count and density were determined using methods described by Biswas et al, (2002) and Magbagbeola et al (2000) respectively. Sperm morphology was determined by the method described by Amelar and Dublin (1978) while the sperm viscosity was evaluated using the method described by Barrat and John (1998). Semen pH was determined by dipping pH paper into the sample and checking the resultant colour against known standards. (Comhaire and Vermealen, 1995). RESULTS: Table 1: Effect of diet incorporating Mucuna rostrata seed meal on some characteristics of male rat sperm: Parameter Control Group 1 Group II 10% 20% Supplementation Supplementation Colour Whitish Whitish Grey Whitish Grey Sperm count/ml Grey 38.00x10 6 39.00x10 6 35.00x10 6 Sperm Motility (%) Active 80 78 70 Sluggish 20 20 25 Dead 0 2 5 Sperm Murphology (%) 72.03 + 0.02 73.30 + 0.03 71.00 + 0.03 Sperm density (10 6 /ml) 65.10 + 0.02 67.10 + 2.00 66.20 + 1.00 Semen pH 7.50 + 0.03 7.50 + 0.02 7.50 + 0.02 Viscosity/Turbidity Normal Normal Normal Progression Good Good Good International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 382 Figure 1: Photomicrograph of testes of rats fed with Mucuna rostrata supplemented meal for 24 days at 10% (b) and 20% (c) compared with control (a) Sd: Spermatids; Sc = Spermatocysts; Sn = Spermatogonia L: Leydig; Sz= Spermatozoa; S = Seminiferous tubule DISCUSSION: The results of the effect of diet incorporating Mucuna rostrata seed meal on some sperm characteristics as reported in Table 1 shows that the sperm colour of the 3 groups of animals was whitish grey. Viscosity and turbidity remained “normal” for the control and test groups. This indicates that the sample had no effect on the parameters. Dean & John (2006) reported that most semen is white, but that grey or even yellowish semen can be normal as well. The non significant effect of the meal on viscosity and turbidity of the semen may be an indication that the Mucuna supplements did not cause abnormal liquefaction. Comhaire and Vermeulen (1995) warned that the semen that is too thick and does not liquefy is due to infection in seminal vesicle or prostate. In the study, the semen pH (7.50) was within the WHO (1993) range for normal semen; an indication that there was no remarkable malfunction which would have resulted from decreased secretion of acidic products by the prostate. Dua & Valdya (1996) reported that abnormal citrate (c) International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 383 secretion, incomplete ejaculation or occlusion of the seminal vesicles could alter the semen pH to a more acidic range; this in turn will affect the sperm cells in their bid to overcome the hostile environment of the female reproductive canal and denature the DNA in the sperm. The motility status of the rat sperms in the test groups compared to the control was not significantly different since only 50% of the cells were dead at 20% level of supplementation. The Environmental Health Perspective, EPH, (2005) alerted several factors which affect sperm motility to include too much heat, long periods of sexual activity, poor diet, chemicals, toxins and prescribed drugs. For this study the 70% motility at 20% level of supplementation did not cause pronounced harm to the sperm cells since the motility rate was within the acceptance 50% WHO (1992) recommendation. No wonder Fertil et al (2007) reported that dosecontrolled treatment of Mucuna pruriens herbs on semen profile and biochemical parameters of infertile men increased sperm concentration and motility; and recommended its judicious use in treatment of digozoospermic patients. The sperm count for the control and test groups was impressive because they were within the acceptable value of 20x10 6 – 40x10 6 /ml for healthy mature male (WHO, 1992). The implication of this result is that the diet did not adversely affect the semen volume or pose any problem to the seminal vesicle or the prostate gland. Guillet & Guillet, (2005) maintained that abnormal semen volume (<10million) signal problem in the seminal vesicle or prostate gland. That there were non significant changes on the morphology of the rat sperm of the test groups compared to control suggest that the sample did not pose adverse effect at those level of supplementation. Sperm morphology like motility depends on the consistency of seminal fluid (Dean, 2006), Hence for conception to occur seminal fluid should be of correct consistency for sperm to have maximum motility and ideal morphology (WHO, 1992). The result of incorporating roasted Mucuna rostrata seed meal in the rat’s diet on their testes International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 384 histology as showed in Fig. 1 had mild effects. At magnification of 10, the testes of the control group clearly showed seminiferous tubule(s) and their epithelia as well as the interstitial cells of the Leydig (L) within the spaces adjoining these tubules. The spermatogonia (Sn), spermatocytes (Sc) and spermatids (Sd) were also seen within the tubules as round oval shaped cells within the seminiferous tubules. The impressions for animals fed with 10% supplementation were those of hyper-cellular testes consisting of normal spermatogenic cells which explains that at that level of supplementation, the sample may not only increase spermatocytosis, but may improve male infertility if it is due to hypospermia or azospermic The fact that the plate of the testes of test group 2 rats showed marked enlargement of the interstitial spaces and cells within it implies that the spermatogenic series of the cells were hyper plastic; hence increased spermatoceptosis (Gonzales & Vilena, 2001). The plate also showed evidence of hypertrophy and hyperplasia of the Leydig cells, hyperplasia of the spermatogenesis (transformation of spermatids to spermatozoa). Thus there was increased spermatogenesis at 20% level of supplementation. Conclusion: Mucuna rostrata has demonstrated its potential on semen profile of male albino rats. The results of this research also shows that Mucuna rostrata seed can be inscribed as fertility enhancer because of its potential fertility promoting and sperm producing effect (being able to improve sperm count, spermatogenesis and motility. Supplementation of the diet with 10% and 20% Mucuna rostrata sample had significant effect on the histological features of the rats testes as evident in progressive increase in both size and number of the Leydig cells of all the groups. There was also profound increase in the mucosal thickness and that of epithelial folds from groups 1-3. This suggest that Mucuna meal may boast the activity of the spermatogenic series of cells, produce a corresponding increase in plasma testosterone cause an increase in the production and maturation of sperm cells and therefore could be useful in improving male fertility. International Journal of Advanced Scientific and Technical Research ISSN 2249-9954 Available online on http://www.rspublication.com/ijst/index.html volume 15, No. 5, 2025 DOI: 10.5281/zenodo.17368717 Original Article ©2025 RS Publication, [email protected] 385 REFERENCES Amelar, R., and Dublin, L. (1978). Serum Analysis: An Official Technical. Urology, 2,606611. Barrat, C. R. and John, C. J. (1998). Diagnostic Tools in Male Inferlility. Journal of Human Reproduction, 13, 15-61. Biswas, D., Barri, F., Shamsuddin, M. and Rahman, M. (2002) Determination of Glycerol Percentages, for preserving the Black Benga Buck Spermatozoa for Long Time. 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