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Effect of Semecarpus anacardium fruits on reproductive function of male albino rats

Arti Sharma1, Pramod Kumar Verma2, V.P. Dixit2

1Department of Zoology, Raj Rishi Colloge, Alwar (Raj) 301001, India
2Reproductive Physiology Section, Department of Zoology, University of Rajasthan, Jaipur-302 004, India

Asian J Androl 2003 Jun; 5: 121-124             

Keywords: Semecarpus anacardium; spermatogenesis; androgen; spermatocytes; Leydig cells

Aim: To evaluate the effect of an ethanolic extract of Semecarpus anacardium fruits on spermatogenesis in albino rats. Methods: Male albino rats were fed with a 50 % ethanolic extract of Semecarpus anacardium fruit at 100 mg.kg-1.day-1, 200 mg.kg-1.day-1 and 300 mg.kg-1.day-1 for 60 days. Fertility test was performed after 60 days of treatment. Sperm motility and density were observed in the cauda epididymis. Biochemical and histological analyses of the blood and reproductive organs were done. Recovery of fertility was followed to evaluate the reversibility of drug action. Results: S. anacardium fruit extract administration resulted in spermatogenic arrest in albino rats. The sperm motility and density was reduced significantly. The RBC and WBC counts, haemoglobin, haematocrit, blood sugar and urea were found to be within the normal range in the whole blood. The protein, cholesterol and glycogen in the testes and the fructose in the seminal vesicle were significantly decreased after the treatment. The fruit extract feeding caused marked reduction in the number of primary spermatocytes, secondary spermatocytes and spermatids. The number of mature Leydig cells was also decreased and degenerating cells increased proportionately. Conclusion: S. anacardium fruit extract causes spermatogenic arrest in albino rats.

1 Introduction

Much attempts have been made to search for male antifertility agent from the plants [1, 2]. Semecarpus anacardium belongs to family Anacardiaceae and is commonly known as Bhilawa in Hindi. The fruits of plants are largely used in Ayurvedic system of medicine for various ailments, particularly alimentary tract and certain dermatologic conditions. It has beneficial action on heart, blood pressure, respiration, cancer and neurological disorders [3-5]. The present investigation was designed to determine the effect of an ethanolic extract of Semecarpus anacardium fruits on spermatogenesis in male rats.

2 Materials and methods

2.1 Plant extraction

Authenticated fruits of Semecarpus anacardium were obtained from the National Institute of Ayurveda, Jaipur. The nutshell tops were removed, the nutshells were powdered and then boiled in distilled water for 8 h~10 h to reduce pungency. The powder was air dried and defatted with petroleum ether. Defatted material (100 g) was soaked in 300 mL of 70 % methanol for 24 h and then filtered. The solvent was removed by vacuum distillation and dried to obtained solid mass (9.7 g solid mass was obtained from 100 g of nutshell). It was washed with petroleum ether, benzene, chloroform and acetone. A pale yellow crude extract was obtained.

2.2 Experimental procedure

Male albino rats weighing 175 g~225 g were divided into four groups of 10 animals each. Group 1 served as the control and received the vehicle, Group 2 received the extract at a dose of 100 mg.kg-1.day-1, Group 3 at 200 mg.kg-1.day-1 and Group 4 at 300 mg.kg-1.day-1. The animals were housed in plastic cages under standardized conditions (12 h light/ 12 h dark; 25 3 and 35 %~ 60 % humidity). They were maintained on standard rat pellet diet and water ad libitum. The plant extract was freshly dissolved in distilled water and administered orally to the animals every morning for 60 days. Mating test was performed with proestrous females after 55 days of treatment. Vaginal smears were checked every morning. The implantation sites were checked on day 16 by laparotomy. The male rats were sacrificed on the next day after the last dosing. Blood was collected through cardiac puncture and the reproductive and the vital organs were dissected out and weighed.

2.3 Sperm analysis

Sperm motility and density were assessed in caudal epididymis by the method of Prasad et al. [6].

2.4 Blood analysis

Whole blood was analyzed for the RBC and WBC counts, haemoglobin, haematocrit, sugar and urea.

2.5 Tissue biochemistry

The testis from each rat was kept at -20 until assayed for the cholesterol, glycogen, protein and sialic acid. Fructose was determined in the seminal vesicle.

2.6 Histological analysis

Testes were fixed in Bouin's fluid, passed through ascending series of ethanol and then through xylene, and embedded in paraffin wax. Tissues were sectioned at 5 mm and stained with haematoxylene and eosin.

The evaluation of the cell population was based on the calculations made for each cell type per cross section of the seminiferous tubule. The Sertoli cells, spermatogonia, primary spermatocytes (preleptotene and pachytene), secondary spermatocytes and round spermatids were counted under 100 magnification. These crude counts were corrected by using Abercrombie's formula [7]. Mature and degenerating Leydig cells were also counted. Leydig cell nuclear area was measured at 800 magnification.

2.7 Recovery observation

After completion of the treatment, half of the animals were kept for recovery observation for a period of 60 days. Their fertility was again accessed by mating test. The mated females examined on day 16 of pregnancy for implantation sites.

2.8 Statistical analysis

Data were expressed in meanSEM and the significance of difference was analyzed by the student's t-test.

3 Results

3.1 Fertility and sperm analysis

None of the females mated with the male rats treated with S.anacardium fruit extract at various dose levels was pregnant. Sperm motility was significantly decreased by 44.07 % and 32.49 % at 200 mg and 300 mg dose levels, respectively, whereas no significant change was observed at 100 mg. Significant reduction (P<0.01) was observed in sperm density at all the three dose levels (Table 1).

Table 1. Fertility and sperm parameters. cP<0.01, compared with controls.



Sperm Motility

Sperm Density

1 (control)
















3.2 Blood analysis

Blood variables i.e. RBC, WBC, haemoglobin, haematocrit, blood sugar and urea were found within the normal ranges after S. anacardium treatment.

3.3 Biochemical finding

Glycogen, cholesterol, protein and sialic acid contents were decreased significantly (P<0.01) in testes of animals treated with the plant extract in comparison to the controls. There was a 50 % reduction in seminal vesicle fructose at 100 mg and 300 mg doses and a 52 % reduction at 100 mg dose in comparison to controls (Table 2).

Table 2. Tissue biochemistry (in mg/g). cP<0.01, compared with controls.



Seminal Vesicle




Sialic Acid

1 (control)
























3.4 Histological observation

S. anacardium fruit crude extract caused depletions in various germinal cell and Leydig cell components at 100 mg, 200 mg and 300 mg dose levels. The number of spermatogonia was decreased by 52.9 %, 19.3 % and 29.8 % at 100 mg, 200 mg and 300 mg doses, respectively. Primary spermatocytes (preleptotene and pachytene), secondary spermatocytes and spermatids were reduced significantly in all the treatment groups (Table 3). Increased number of abnormal seminiferous tubules was observed at all the dose levels. There were 29.4 %, 27.4 % and 47.9 % reductions in the Leydig cell nuclear area at 100 mg, 200 mg and 300 mg doses, respectively. The number of mature Leydig cells was decreased, and the degenerating cells increased (P< 0.01) in comparison to controls (Table 4).

Table 3. Testicular cells. bP<0.05, cP<0.01, compared with controls.


Preleptotene spermatocytes

Pachytene spermatocytes

Secondary spermatocytes

Round spermatids

1 (control)




















Table 4. Seminiferous tubules and Leydig cells. cP<0.01, compared with controls.


Tubules (%)

Stage 19
Spermatids (%)

Leydig cell
Area (µm2)

Leydig cell
Nuclear area (µm2)

Mature Leydig
cells (%)

Leydig cells (%)

Fibroblast like
cells (%)

1 (control)
































3.5 Recovery

All treated animals became fertile after 60 days of drug withdrawal. Females mated with these males delivered healthy litters.

4 Discussion

Semecarpus anacardium extract feeding caused antispermatogenic effect evidenced by reduction in numbers of spermatogenic cells and spermatozoa. Reduction in sperm density in cauda epididymides may be due to changes in the androgen metabolism. The principal cells of epididymis synthesize proteins, which have important role in maturation of spermatozoa [8]. Alterations in the secretion and function of these proteins impared sperm maturation. The epididymal protein was reduced in the present study as with other plant materials, including Sarcostemma acidum [9] and Barleria prionitis [10] in rats and Mentha arvensis in mice [11]. Low fructose concentration may be another cause of reduction in sperm motility as seminal fructose provided energy for sperm motility. The reduction in the glycogen level in testes may be due to interference in glucose metabolism. The reduced glycogen level was correlated with diminished number of post meiotic germ cells, which were supposed to be the site of glucose metabolism. The reduced glycogen level could affect protein synthesis, because protein synthesis in spermatogenic cells was dependent on glucose [12]. Gupta et al. [10] also observed reduced glycogen and protein levels in testes of Barleria prionitis root extract treated rats. Decreased number of spermatozoa or reduced androgen production may affect the level of sialic acid in testes. The reduced sialic acid content might alter the structural integrity of acrosomal membrane which ultimately affects the metabolism, motility and fertilizing capacity of spermatozoa [13]. Semecarpus anacardium fruit extract feeding caused impairment of Leydig cell function, which was evidenced by reduced Leydig cell area and nuclear dimensions and fewer number of mature Leydig cells. The atrophic state of Leydig cells in the testes of treated animals may be due to declined LH secretion [14, 15]. Differentiation of primordial germ cells into spermatogonia and subsequent appearance of spermatogenic cycle are under the control of gonadotropin and testosterone, such control being possibly mediated by Sertoli cells [16], which regulate cell cycle kinetics and influence both spermatogonia and preleptotene spermatocytes [17]. The reduction in number of secondary spermatocytes and spermatids reflected non-availability of ABP from Sertoli cells [18]. ABP is required to maintain intra-testicular androgen concentration and transformation of advance stages of germ cells. Meiotic and post-meiotic germ cells were highly sensitive to androgen concentration [19] and the alteration in androgen level in testes may affect the transformation of spermatocytes to spermatids. The blood parameters remained within the normal range after Seme-carpus anacardium administration indicating non-toxic nature of the plant.


The authors are indebted to Indian Council of Medical Research, New Delhi, India for financial support for this study.


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Correspondence to: Dr. Arti Sharma, 1389, Baba Harish Chandra Marg, Bhoora Tiba, 3rd Cross Road, Jaipur (Rajasthan) 302 001 India.
E-mail: artiz@rediffmail.com , vivek@cswri.raj.nic.in
Received 2003-01-10   Accepted 2003-03-07



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