Buchholzia coriacea (wonderful kola) seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis in Wistar rats

The methanolic extract of Buchholzia coriacea seeds (MEBC) has been reported to induce male reproductive toxicity by decreasing sperm parameters and fertility index. To elucidate the possible mechanism(s), the effects of graded doses of MEBC on sex hormones and sperm profile were investigated in this study. The MEBC (e.g., 50, 200, 400, and 600 mg/kg) was administered daily (p.o.) to male Wistar rats for 6 weeks, while a concurrent control group received distilled water (vehicle). Then, the animals were sacrificed under sodium pentobarbital anaesthesia. Weights of organs were recorded, and the sperm profile was determined microscopically. Testosterone, luteinizing hormone (LH), and follicle stimulating hormone (FSH) were assayed from the obtained serum using the ELISA technique. Sperm motility was significantly reduced by MEBC (i.e., 50 and 200 mg/kg), and sperm count reduced in all treated groups in a dose-dependent manner compared with that of the control. Serum testosterone, LH, and FSH decreased in treated rats. A histopathological examination of testes showed a considerable depletion and necrosis of the epithelium of seminiferous tubules. The result suggests that Buchholzia coriacea seeds induce male reproductive toxicity by suppressing the pituitary-gonadal axis.

Fertility Enhancing Potency of Omega-3 Fatty Acids in Male Rats

Abstract Cases of some grave side effects of some allopathic medicines used in treatment of infertility has caused a global need for alternatives with minimal or no side effect, hence the demand to evaluate the fertility enhancing potential of omega-3 fatty acids (O3FA) in male rats. This study evaluated the fertility enhancing potential of omega-3 fatty acids (O3FA) in male rats. Seventy-two sexually mature male albino rats 11-13 weeks of age, weighing between 171 - 180 g were assigned into six groups (I - VI) fed graded doses of O3FA. Administration of O3FA lasted twenty-eight days at 48 hour intervals. At the end of the treatment, organosomatic index of testes, testicular and epididymis sperm cells counts and testicular histology were assessed following standard methods. The actual and relative testicular weights, testicular and epididymis sperm counts of all O3FA treated rats were significantly increased (p < 0.05) when compared with the control group rats. The photomicrographs of testes in O3FA treated rats showed normal spermatogonial cell layers and active spermatogenesis with appearance of spermatids in the lumen of some tubules. The findings of this study depicts that O3FA possesses the potency of enhancing various fertility indices in male rats with regards to absolute and relative testicular weights as well as sperm counts.

Aggravation of cyclophosphamide-induced reproductive toxicity in mice by aqueous extract of Aegle marmelos (L. )

ABSTRACT Aegle marmelos (L.) (Rutaceae) commonly known as bael is an important medicinal fruit tree. The present study focused on the effects of aqueous extract of Aegle marmelos (AEAM) on the testis and sperm characteristics induced by cyclophosphamide (CPA) in mice. Thirty six adult Parke's strain mice were divided into six groups: group I given only distilled water (control); group II administered with AEAM alone once in a week for five weeks; group III administered with CPA (200 mg/kg b.w., intraperitoneally) once in a week for five weeks and group IV-VI CPA along with AEAM (400, 500 and 600 mg/kg b.w., orally). CPA was found to reduce gonadosomatic index (GSI), sperm counts, motility, viability, antioxidant activities and induced histopathological changes of testis. In the group administered AEAM with CPA an exacerbation of sperm count, motility and viability of the cauda epididymis, GSI, antioxidant activities and architecture of testis was observed. The results suggest that the administration of AEAM may aggravate CPA-induced reproductive toxicity. It may be helpful in preparation of natural male contraceptives.

Tissue perfusion-controlled guided biopsies are essential for the outcome of testicular sperm extraction.

OBJECTIVE: To determine if there are areas of major and minor perfusion in a single testicle, and if the quality and quantity of sperm are correlated with the level of perfusion, we collected testicular tissue from areas with different levels of perfusion. DESIGN: Controlled clinical study. SETTING: Consecutive patients with azoospermia. PATIENT(S): Patients with azoospermia undergoing testicular sperm extraction (TESE) biopsy for the retrieval of sperm to be used in an assisted reproduction program. INTERVENTION(S): Perfusion mapping was performed with the use of color Doppler ultrasound. Areas with different levels of perfusion were marked with needles. After incision with radiofrequency cutting, the exposed tissue was examined with a laser Doppler flowmeter, and biopsies were taken for TESE and histology. Sperm were analyzed using World Health Organization criteria, and prepared for intracytoplasmic sperm injection (ICSI). MAIN OUTCOME MEASURE(S): Correlation of sperm quality and quantity in testicular-tissue biopsies, with tissue-perfusion units (TPU) measured by laser Doppler flowmeter. RESULT(S): From 40 biopsies taken from 20 testicles of 12 patients, tissue was analyzed for sperm quality and quantity. Sperm quality was highest in areas of high tissue perfusion. In areas of 70 TPU, 72.3% progressive sperm were detected, whereas in areas of 10 TPU, only 13.3% progressive sperm and elevated numbers of precursor cells could be observed. The number of motile sperm isolated from tissue samples correlated well with the intensity of tissue perfusion. CONCLUSION(S): We have shown for the first time that in patients suffering from azoospermia, sperm quality and quantity depend on tissue perfusion within the testicle.

Influence of sample preparation, staining procedure and analysis conditions on bull sperm head morphometry using the morphology analyser integrated visual optical system.

The importance of standardizing the procedures of sample and slide preparation for computer-assisted morphologic analysis has been emphasized in human and veterinary andrology. The purpose of this study was to optimize slide preparation (dilution grade and sperm washing), staining procedures and analysis conditions (colour of light source and objective magnification) for the morphometric analysis of bull spermatozoa using the Hamilton Thorne morphology analyzer integrated visual optical system (IVOS). For experiment 1, one ejaculate was collected from one bull and diluted to 200,000-300,000 spermatozoa/microl. Slides were prepared and stained using seven different procedures: rapid Papanicolaou (PAP), rapid Papanicolaou with prolonged staining times (PAP+). Diff-Quik (DIF), haematoxylin (HEM). Farelly (FAR), Spermac (SPER) and the modified GZIN (MGZIN) staining. All slides were analysed using a Hamilton Thorne Morphology Analyser IVOS equipped alternatively with a red, green or blue light source, and a 40x or 100x oil immersion objective. Recognition and digitization errors as well as morphometric parameters were determined. The IVOS was unable to detect DIF-stained spermatozoa. The GZIN and the SPER staining as well as the blue light source led to unsatisfactory results. Among the staining methods examined, the FAR, HEM, PAP+, and PAP staining, preferably in combination with the green light source, and the 40x objective yielded optimal results concerning sperm recognition and digitization. The 100x objective did not allow reliable analysis of the sperm heads because of a frequently appearing digitization error. For experiment 2, three ejaculates were collected from each of three bulls and diluted to five dilution grades (100 000-500 000 spermatozoa/microl). An aliquot of each dilution grade was washed additionally. The percentage of correctly digitized sperm heads decreased with increasing spermatozoal concentration. However, the evaluation speed increased. The range of 200 000-300 000 spermatozoa/microl appeared to be a reasonable compromise for both criteria. Sperm washing failed to further improve the analysis results. Sperm head dimensions were influenced significantly by all variations of the methods in both experiments. In conclusion, using the proposed methods, the IVOS allows precise and reliable morphometric analyses of bull spermatozoa. The consistent application of these procedures may lead to an inter-laboratory standardization and to further establishment of generally accepted morphometric criteria used in human andrology (e.g. World Health Organisation or strict criteria).

Espermograma: indicaciones e interpretación

La evaluación y tratamiento integral del varón infértil es requisito indispensable para el logro de embarazos, No siempre se consigue conocer la cuasa de la infertilidad masculina. La evaluación convencional y funcional del semen según los criterios actualizados de la Organización Mundial de la Salud nos premite dar un diagnóstico y tratamiento más adecuado y científicos a la pareja infértil. Estos criterios se han constituido en la referencia estándar para todos los laboratorios que analizan muestras de semen humano. Las técnicas de reproducción asistida permiten a los hombres infértiles, en un procentaje muy alto, realizar sus deseos de paternidad biológica. En este módulo se definen las características normales y anormales del semen y se describen los diferentes procedimientos que componen un espermograma completo; además, se presentanlas indicaciones y la interpretacion de sus resultados.

Sperm function and choice of preparation media: comparison of Percoll and Accudenz discontinuous density gradients.

We compared the sperm populations prepared by Accudenz (35-65%) and Percoll (40-80%) density gradients in 21 normospermic specimens (concentration, 53.6 +/- 3.8 x 10(6) sperm/ml; motility, 44.5 +/- 3.5%). Accudenz facilitated a higher recovery of sperm and motile sperm (68.4 +/- 6.6% vs. 49.3 +/- 4.9%, P < 0.001, and 87.8 +/- 4.1% vs. 77.8 +/- 3.7%, P < 0.01, respectively). Sperm motility was lower in the Accudenz compared to the Percoll pellets; thus the values of total motile sperm recovered were not different (17.1 +/- 2.4 vs. 15 + /- 1 2.2 x 10(6) sperm/ml). The long term retention of sperm motility was substantially improved in Accudenz (at 24 hours, 34.9 +/- 2.8% vs. 26.3 +/- 1.5%; 60% vs. 40% of the initial motility, P < 0.001), and the Accudenz vs. Percoll samples also exhibited a higher retention of total motile sperm (at 24 hours, 9.8 +/-.2 vs. 6.1 = 0.5 x 10(6) motile sperm/ml, P < 0.05). The sperm motility index, a multiple of velocity and motility in the sample that reflects the efficiency of the sperm population in sperm-oocyte interaction, was 75% higher in the Accudenz samples at 24 hours (3.6 +/- 0.4 vs. 2.1 +/- 0.2, mu m/second, P < 0.01). Sperm cellular maturity by the creatine phosphokinase (CK) activity and CK-M to CK-B isoform ratio parameters (in the original samples 0.14 +/- 0.02 lU CK/100 x 10(6) sperm and 57.9 +/- 3.7%, respectively) were improved in both the Accudenz and Percoll pellets (P < 0.001), with no difference between the two sperm fractions. Sperm activation status monitored by chlortetracycline fluorescence indicated that after 4 hours of incubation the incidence of fully acrosome-reacted spermatozoa in the Accudenz versus Percoll pellets was 6.2 +/- 0.3% versus 13.1 +/- 1.0% (P < 0.001), a 100% increase in Percoll. We can conclude that Accudenz yields a higher concentration of motile spermatozoa, with improved retention of motility, velocity, and acrosomal integrity and without an increase of sperm with diminished cellular maturity. Thus, in sperm preparation for intrauterine insemination, in which the timing of ovulation and insemination frequently do not correspond, Accudenz-prepared sperm, with a better retention of motility/velocity and acrosomal integrity and with a consequential higher resistance to activation by the female reproductive tract, are expected to be more effective.