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This study was conducted to evaluate the adverse effects of the anabolic steroid, boldenone undecylenate BOL on reproductive functions of male rabbits. Thirty white New Zealand mature male rabbits were divided into three groups 10 rabbits each. Group A rabbits served as a control group.
Group B rabbits received 4. Group C rabbits received 8. Rabbits were injected intramuscularly twice weekly for two months. BOL had no significant effect on the bwt and bwt gain. Testes and epididymis weights were decreased significantly in the BOL-treated groups. BOL caused significant reduction in serum testosterone level, seminal volume, sperm motility, and sperm count.
No abnormalities were detected in the sperm morphology of the BOL-treated groups. Histopathological alterations in the testes and epididymis were marked in the group C rabbits.
These results indicate that administration of BOL exerts a significant harmful effect on the reproductive functions of male rabbits. Anabolic androgenic steroids AAS are synthetic derivatives of the male testosterone hormone that have been modified to improve their anabolic rather than androgenic activity Shahidi Boldenone BOL is an anabolic steroid that differs from testosterone only by one double bond at the 1- position Stolker et al.
It is used mainly as undecylenate ester by bodybuilders and is administered illegally to racing horses. However, it is used as a growth promotor on farms improving the growth and feed conversion of cattle; it may be abused to achieve more efficient meat production Gryglik et al. In developing countries with rapid growth of population, like Egypt, the demand for edible protein exceeds the supply and the gap is expanded.
Meat from animals, including from rabbits, provides a valuable and palatable source of protein. We found BOL to be used heavily in Egypt, not only in the field of animal production, but also by athletes and bodybuilders. BOL increases muscle size owing to promotion of positive nitrogen balance by stimulating protein production and reducing protein destruction, as well as causing retention of body water, nitrogen, sodium, potassium and calcium ions Forbes ; Mooradian et al.
Despite these restrictions, AAS are easily obtained. The abuse of AAS can lead to serious and irreversible organ damage Maravelias et al.
Among the most common adverse effects of AAS that have been described are reduced fertility Dohle et al. There have been relatively few studies which have investigated the detrimental effects of BOL administration on male function. Hence, this study was performed to determine the effects of high-dose administration of BOL on body weight bwt , reproductive organ weight, semen characteristics, serum testosterone levels and histopathological features of the reproductive organs of mature male rabbits.
Thirty white New Zealand mature male rabbits, 9—9. Fed pelleted commercial feed Ibex Co. Rabbits in all groups received humane care in compliance with the animal care guidelines of the National Institute of Health, and the local ethical committee approved this study. Rabbits were divided into three groups 10 rabbits each.
Group A rabbits served as control group and received 0. Group B received 4. All groups were injected intramuscularly twice weekly for 2 months. Rabbits were ranked by restricted randomization procedures that approximately equalized the initial bwts among the different groups. They were then weighed weekly until the end of the experiment.
Final bwt was recorded, and weight gain was calculated. All rabbits were killed at the end of the experiment. After dissection, the testes, epididymis and prostate glands were removed, grossly examined and weighed. The index weight I. Ejaculates were collected from each male rabbit prior to the treatment, after one month of treatment and at the end of the experiment with a rabbit artificial vagina.
Each buck was conditioned to react with the artificial vagina as described by Breddman et al. Each male was allowed a false mounting for teasing prior to the actual mounting. The semen was evaluated immediately after collection for the following criteria:. Blood was collected from the ear vein of each rabbit before euthanasia. Serum was separated for assessment of the total serum testosterone according to Demetriou using solid-phase radioimmunoassay RIA kits.
This assay was based on presence of a testosterone-specific antibody immobilized to the wall of the polypropylene tube. At the end of the experiment, rabbits were necropsied. Data are presented as means plus or minus the standard error.
The initial bwt of all groups was equalized approximately. Treatment with BOL had no significant effect on the final bwt and the bwt gain of the treated groups compared with the control group Table 1. This reduction was marked in the group C. No significant changes were found in the index weight of the prostates. This reduction was prominent in the group C Table 2. Effect of BOL on reproductive organs weights and serum testosterone levels of male rabbits.
The sperm characteristics of the treated groups were not changed at the first two time points of semen collection compared with the control group Table 3. No significant abnormalities in the sperm morphology were found in all treated groups compared with the control group Table 3.
Histopathological findings of testes, epididymis and prostate gland were evaluated under light microscopy. Incidence and severity of lesions in BOL-treated groups are summarized in Table 4. Testes of the control mature rabbits had normal histoarchitecture, and were composed of uniform, well-organized seminiferous tubules with complete spermatogenesis and interstitial connective tissue Figure 2a.
Testes of group B rabbits showed degenerative changes that were characterized by small, disorganized seminiferous tubules with irregular basement membrane and decreased spermatogenesis. Moreover, there was vacuolar degeneration of the germinal epithelium and Sertoli cells. Lumina of the majority of seminiferous tubules contained sloughed germinal epithelial cells and giant cell formations Figure 2b. Some tubules showed coagulative necrosis with hyalinized luminal contents.
Testicular sections of group C rabbits exhibited marked small-sized, disorganized seminiferous tubules with marked thickened hyalinized basement membrane Figure 2c,d. Vacuolation of spermatogonia and Sertoli cells was seen. There was obvious cessation of spermatogenesis: Also, some tubules had sloughed germinal epithelial cells within their lumina. In the interstitium, there was marked thickening due to increased by fibrous connective tissue. Photomicrograph of rabbit testis stained with HE: Control mature rabbits showed normal epididymal histological architecture with normal sperm density Figure 3a,b.
In group B rabbits some of the epididymal ductules were empty of mature spermatozoa, and others had low density of spermatozoa and sloughed germ cells in their lumina Figure 3c,d.
Epididymal ductules of group C rabbits were free from mature spermatozoa, and some cauda epididymal ductules contained sloughed germ cells Figure 3e,f.
Photomicrograph of rabbit epididymis stained with HE. Caput epididymis c , cauda epididymis d of a rabbit that received 4. Caput epididymis e , cauda epididymis f of a rabbit that received 8.
The prostate of the control rabbits was histologically normal Figure 4a. No detectable changes were noticed in both treated groups apart from some moderate tubular dilatation Figure 4b,c. Photomicrograph of rabbit prostate stained with HE: Few papers have studied the effect of high dose BOL treatment on male reproductive function. Consequently, this study was performed to evaluate the effects of BOL on bwt, bwt gain, reproductive organ weight, serum testosterone level, semen analysis and sperm characteristics and histopathology of reproductive organs of mature male rabbits.
Our study revealed that treatment with BOL had no significant effect on the final bwt and the bwt gain of the treated groups compared with the control group. Similar results have been reported in horses Maher et al. The index weight of testes and epididymes was decreased significantly in the BOL-treated groups, particularly in group C compared with the control group. This result was parallel with the significant reduction in serum testosterone level in these groups compared with the control group.
In contrast no significant changes were found in the index weight of the prostates. Concerning semen quality, at the end of the experiment ejaculate volume, sperm motility and sperm count of BOL-treated rabbits showed a significant reduction, particularly in group C. These results were similar to those reported in stallions by Squires et al. However, no significant changes were detected in sperm abnormalities. Parallel to these findings, the testes of BOL-treated rabbits exhibited different histopathological changes which were more marked in group C.
These changes manifested as shrunken, disorganized seminiferous tubules with marked thickened hyalinized basement membrane, and vacuolation of spermatogonia and Sertoli cells. Also, there was obvious cessation of spermatogenesis. The majority of seminiferous tubules had single or double cell layers. These findings may be attributed to decreased serum testosterone levels in BOL-treated groups. Testosterone is essential for the attachment of different generations of germ cells in seminiferous tubules.
Exogenous treatment with testosterone or AAS such as BOL are followed by suppression of both gonadotropin-releasing hormone production by the hypothalamus and luteinizing hormone production by pituitary gland and consequently lead to suppression of testicular testosterone production Dohle et al. The testicular lesions were similar to those described by Cannizzo et al. The epididymal lesions reflected the cessation of spermatogenesis particularly in group C.
The prostatic lesions were limited except for some moderate tubular dilatation that may be due to hypersecretion; however, there was no significant increase in the index weight of prostates.
Similar findings were described by Groot and Biolatti who found that BOL induced hypersecretion, hyperplasia and cyst formation in the prostate and bulbourethral gland, with reduced spermatogenesis and enhanced degeneration of testicular germinal epithelium.
Thus in conclusion, this study revealed that AAS, and in particular BOL significant had no major effect on bwt gain but induced a deleterious effect on fertility of male rabbits.