P–008 The diameter of standard seminiferous tubules varies with the different histological patterns in patients with non-obstructive azoosperm

Study question Does the standard, not-dilated seminiferous tubules (STs) diameter vary according with different testis histology patterns in patients with non-obstructive azoospermia (NOA)? Summary answer The standard STs diameter differed significantly among cases with different testis histology: hypospermatogenesis (HYPO) had the highest STs diameter compared to the other histological subgroups. What is known already During microdissection testicular sperm extraction (mTESE), the identification of dilated STs, as subjectively evaluated at high magnification by comparing their apparent caliber with that of the surroundings, is crucial to identify residual foci of intact spermatogenesis and to retrieve sperm. Previous studies have demonstrated that dilated STs contain sperm in most cases, but it is not clear why in some cases an apparent normal tubular caliber does not correspond to spermatogenesis integrity. Aim of the present study was to assess whether different histology patterns could affect the STs diameter. Study design, size, duration We retrospectively evaluated 168 patients with NOA undergoing unilateral (N = 91) or bilateral (N = 77) mTESE from 2018 through 2019. One or more biopsy samples representative of the overall appearance of the testicular parenchyma were taken from one (for unilateral mTESE) or both testes (for bilateral mTESE), was fixed in Bouin’s solution and sent to the pathologist. Histological analysis was conducted by the same experienced pathologist, who examined at least 100 different tubule sections per biopsy sample. Participants/materials, setting, methods Each tubule section (N = 100 per sample) was evaluated at 10X magnification with a micrometer to measure the tubule diameter, then the mean ST diameter was computed. The basal membrane (BM) thickening was evaluated in every section, and a score was assigned by multiplying the degree of BM fibrosis (mild=1, moderate=2, severe=3) for the number of sections (e.g. BM score for moderate fibrosis in 50 sections=2x50=100). Leydig cells hyperplasia (LCH), if present, was also annotated. Main results and the role of chance The median + interquartile range STs diameter was 140; 110–185 mm, while the median BM score was 100; 10–150. Sertoli cell only syndrome (SCO) was found in 51.1% of cases, focal SCO (FSCO) in 4,7%, early (EMA) and late (LMA) maturation arrest in 10.2 and 2.73% respectively, HYPO in 26.17% and hyalinosis (HL) in 5% of cases. LCH was found in 46.88% of samples. STs diameter, BM score and LCH differed significantly among the different histological patterns: STs diameter was 125; 100–148 in SCO, 162; 102–187 in FSCO, 130; 100–175 in EMA, 145; 130–195 in LMA, 205; 170–240 in HYPO and 57.5; 42.5–100 in HL. HYPO samples also had the lowest BM score (20;1–100; p < 0.0001) and LCH prevalence (23.8%, p < 0.0001) compared to the other histological subgroups (HS). A multinomial logistic regression for prediction of different histological subgroups was run with STs diameter, BM score and LCH as candidate predictors: the model explained the 29% of variability of the outcome and correctly classified 69% of cases. STs diameter significantly predicted FSCO (RR 1.02, 95% CI 1.0–1.04), LMA (RR 1.02, 95% CI1–1.04) and HYPO (1.03, 95%CI 1.02–1.04), while BM score significantly predicted HL (RR 1.07, 95%CI 1.02–1.13). Limitations, reasons for caution The STs were carefully cut before extraction in order to preserve their structural integrity, however the accuracy of such a method of estimating the STs diameter needs to be assessed by further studies. Wider implications of the findings: The identification of dilated STs remains the best strategy to retrieve sperm by mTESE, since larger STs diameters are mostly associated with the more favorable histological patterns. However, dilated STs may be also found in cases with LMA, which explains why in some cases dilated STs do not contain sper. Trial registration number Not applicable

Marsilea crenata ethanol extract prevents monosodium glutamate adverse effects on the serum levels of reproductive hormones, sperm quality, and testis histology in male rats

Background and Aim: Marsilea crenata is an aquatic plant that contains high antioxidants level and could prevent cell damages caused by free radicals. The present study aimed to investigate the effect of M. crenata ethanol extract on luteinizing hormone (LH), testosterone levels, sperm quality, and testis histology of adult male rats induced by monosodium glutamate (MSG). Materials and Methods: This study randomly divided 48 male rats into eight groups (n=6): control group; MSG group (4 mg/g body weight [b.w.] for 30 days); MS1, MS2, and MS3 groups (4 mg/g b.w. MSG and M. crenata ethanol extract at dose 0.216, 0.432, and 0.648 mg/g b.w., respectively, for 30 days); and S1, S2, and S3 groups (M. crenata ethanol extract at dose of 0.216, 0.432, and 0.648 mg/g b.w., respectively, for 30 days). The blood sample was collected on days 0 and 30 to determine the LH and testosterone levels. The animals were dissected on day 30, and the testes were isolated for morphometric, histology (spermatogenic cell number), and malondialdehyde (MDA) examination. Moreover, semen was collected to determine the sperm quality parameter. Results: The LH and testosterone levels significantly increased (p<0.05) after M. crenata administration at all doses. The higher dose of M. crenata ethanol extract demonstrated a high decrease in MDA level in MSG-treated rat testis; increase of spermatogonia, spermatocytes, spermatids, and Leydig cells number; and increase of seminiferous tubular diameter and germinal epithelium thickness. Conclusion: The ethanol extract of M. crenata can improve the levels of LH, testosterone, sperm quality, number of testis morphometric, spermatogenic, and Leydig cells in MSG-treated male rats.

Protective Effects of Aqueous Extract of Persea Americana Leaves Against Secondary Sexual Traits and Histopathological Damages Induced by Antouka Super® in Male Japanese Quails (Coturnix sp)

The control of pesticide’s toxicological properties in the food chain could be a benefit to farmers by increasing animal productivity. The current study’s objective was to evaluate the protective effects of aqueous extract of Persea americana leaves (AEPAL) on secondary sexual traits and testis histology damages induced by Antouka Super® (AS) in male Japanese quail. Sixty male quails of 28 days old and weighing 106-119g were randomly distributed into 5 groups of 12 animals each and daily received for 60 consecutive days one of the treatments: group 1 and 2 received respectively 10 ml of distilled water and 75mg/kg of b.w of AS while groups 3, 4 and 5 in addition to 75mg/kg of b.w of AS, they received respectively 50, 100 and 200 mg/kg of b.w of AEPAL. The sexual behavior characteristics were evaluated during the trial period. At the end of the study, birds were humanly sacrificed, their blood collected for testosterone analysis and the testes removed for teste structure evaluation. Results revealed that exposure to AS significantly decreased testosterone level, time of shouting and appearance of foam, quantity of foam product, the volume and area of the cloacal gland, frequency of mount and frequency of successful mount, as well as degradation of the testes histology. The administration of AEPAL increased these parameters in a dose-dependent manner. Thus, these results demonstrate that AS had a toxic effect on secondary sexual traits and testis histology in male quail which could be alleviated by AEPAL administration at 200mg/kg of b.w.

Colibri® insecticide induces male reproductive toxicity: alleviating effects of Lannea acida (Anacardiaceae) in rats

Background Imidacloprid, a neonicotinoid insecticide, has been associated to severe reproductive toxicity in mammals. Although some preventive measures have been reported, curative strategies are yet to be explored. The present study was designed to investigate the alleviating effects of Lannea acida on the reproductive toxicity of colibri®, a commercial formulation of imidacloprid, in adult male rats. Materials and methods Seventy rats were orally administered with colibri® (22.5 mg/kg, 10 mL/kg) for 14 days and treated for other 14 or 28 days with either aqueous or methanol extracts of L. acida (170 or 340 mg/kg). Control animals were similarly treated with clomiphene citrate or vitamin E. Sexual organ weights, spermatozoa characteristics, sexual hormones, stress markers and testis histology were evaluated at the end of each treatment period. Results Colibri® exposition induced reproductive toxicity marked by a decrease in sex organ weights, spermatozoa count, motility and viability. Colibri® also decreased testosterone, luteinizing hormone, follicle stimulating hormone concentrations and increased testicular oxidative stress. Spermatozoa morphology and testis histology were also severely altered. Similar to clomiphene citrate and vitamin E, treatment with L. acida extracts significantly (p ≤ 0.05–0.001) reversed the above-mentioned damages, especially after 28 days of treatment with aqueous (340 mg/kg) and methanol (170 mg/kg) extracts. Conclusion Present results indicate that L. acida exerts curative effects against colibri®-induced male reproductive toxicity. These results justify the use of this plant as fertility enhancer and suggest that it could be an alternative in the management of pesticide-derived male infertility.

EKSTRAK DAUN SEMBUNG (Blumea balsamifera) MEMPERBAIKI HISTOLOGI TESTIS TIKUS WISTAR YANG DIINDUKSI PAKAN TINGGI LEMAK

Sembung (Blumea balsamifera) Leaf Extract Improves Testis Histology of High-Fat Diet-Induced RatsABSTRACTHigh fat and high cholesterol diet cause hyperlipidemia, leading to various health problems including reproductive health. The purpose of this study was to examine the effect of sembung (Blumea balsamifera) leaf extract on testicular histology profile of high-fat-diet-induced wistar. This research used 16 adult male rats (Rattus norvegicus), aged 3-4 month, weighing 150-200 g, and randomly divided into two groups. Eight rats were treated with distilled water and eight rats were treated with 2 mg/mL B. balsamifera extract. High-fat diet was a 30 days of porcine fat feed. The results showed that the diameter of seminiferous tubules, the number of spermatogenic cells of spermatogonium A, spermatocytes Pakiten and spermatid 16 were increased by giving sembung leaf extract for 50 days (p <0.05). These results suggested that the sembung leaf extract improves testis histology of high-fat diet-induced rats. Keywords: high-fat diet, rat, sembung leaf, seminiferous tubules, spermatogenic cellsABSTRAKMakanan tinggi lemak dan tinggi kolestrol menyebabkan hiperlipidemia yang menimbulkan masalah pada sistem reproduksi. Penelitian ini bertujuan untuk mengetahui manfaat pemberian ekstrak daun sembung (Blumea balsamifera) terhadap profil tubulus seminiferus dan sel-sel spermatogenik pada tikus wistar yang diinduksi pakan tinggi lemak. Sebanyak 16 ekor tikus wistar jantan dewasa (Rattus norvegicus) umur 3-4 bulan, berat 150-200 g, secara random dibagi dua kelompok, yaitu 8 ekor tikus kelompok kontrol (aquades steril) dan 8 ekor tikus kelompok perlakuan (ekstrak daun sembung dosis 2 mg/mL). Tikus diinduksi pakan tinggi lemak berupa lemak babi selama 30 hari. Pada prettest dilakukan pemeriksaan diameter tubulus seminiferus dan sel-sel spermatogenik. Hasil penelitian menunjukkan bahwa rata-rata diameter tubulus seminiferus tikus meningkat signifikan setelah pemberian ekstrak daun sembung. Peningkatan secara signifikan juga diikuti oleh spermatogonium A, spermatosit Pakiten dan spermatid 16 (p<0,05). Dapat disimpulkan bahwa ekstrak daun sembung memperbaiki histologi testis tikus wistar yang diinduksi pakan tinggi lemak.Kata Kunci: daun sembung, pakan tinggi lemak, sel spermatogenik, tikus, tubulus seminiferus