The effect of l-arginine and flaxseed on plasma testosterone concentration, semen quality and some testicular histology parameters in old broiler breeder roosters.

The objective of this study was to investigate the effect of dietary supplementation of l-arginine and flaxseed on testosterone and lipid parameters of serum and semen quality, as well as histological and morphometric parameters of testes in old broiler breeder rooster. Thirty six 52-week-old Ross 308 broiler breeder roosters with similar weights (4900 g ±â€¯210) were used for a 8-weeks trial period in a 3 × 3 factorial arrangement of dietary treatments that three levels of l-arginine (0.52, 0.68 and 0.83%) and flaxseed (0, 2 and 4%) with four replications in each. The ratio of left testicle weight/total testicular weight was significantly higher in A68F0 group. The maximum and minimum seminiferous tubule diameters were recorded for roosters fed no flaxseed and F2 flaxseed, respectively. The roosters fed A68 showed the highest serum testosterone. Seminal volume and sperm concentration, were not significantly affected by l-arginine × flaxseed interaction; however, sperm forward motility were significantly affected by arginine × flaxseed interaction (P < 0.05). A68F0 had significantly greatest tubular differentiation indices (TDI) in all of the treatments. Also, repopulation indices (RI) and spermiogenesis indices (SI) in A68F0 were significantly higher than other groups, but A52F0 and A52F4. However, testis index, testis density, serum lipids (cholesterol, LDL, HDL and HDL: LDL ratio), epithelium height of seminiferous tubules and number of Sertoli cells were not affected by the treatments. It is concluded that dietary supplementation of arginine (0.68%) has positive effects on blood testosterone, semen quality and spermatogenesis index of aged roosters.

[Lipoic acid protects spermatogenesis in male rats with ornidazole-induced oligoasthenozoospermia].

OBJECTIVE: To study the protective effect of lipoic acid (LA) on the spermatogenic function of the male rats with oligoasthenozoospermia induced by ornidazole (ORN). METHODS: Seventy male SD rats were equally randomized into groups A (solvent control: 1 ml 0.5% CMC-Na + 1 ml olive oil), B (low-dose ORN model: 400 mg/kg ORN suspension + 1 ml olive oil), C (low-dose ORN + low-dose LA treatment: 400 mg/kg ORN + 50 mg/kg LA), D (low-dose ORN + high-dose LA treatment: 400 mg/kg ORN + 100 mg/kg LA), E (high-dose ORN model: 800 mg/kg ORN suspension + 1 ml olive oil), F (high-dose ORN + low-dose LA treatment: 800 mg/kg ORN + 50 mg/kg LA), and G (high-dose ORN + high-dose LA treatment: 800 mg/kg ORN + 100 mg/kg LA), and treated respectively for 20 successive days. Then all the rats were sacrificed and the weights of the body, testis, epididymis and seminal vesicle obtained, followed by calculation of the organ index, determination of epididymal sperm concentration and motility, and observation of the histomorphological changes in the testis and epididymis by HE staining. RESULTS: Compared with group A, group E showed significantly decreased body weight (ï¼»117.67 ± 11.53ï¼½ vs ï¼»88.11 ± 12.65ï¼½ g, P < 0.01) and indexes of the testis (ï¼»1.06 ± 0.12ï¼½ vs ï¼»0.65 ± 0.13ï¼½ %, P < 0.01) and epididymis (ï¼»0.21 ± 0.03ï¼½ vs ï¼»0.17 ± 0.01ï¼½ %, P < 0.01). In comparison with group E, group F exhibited remarkable increases in the epididymal index (ï¼»0.17 ± 0.01ï¼½ vs ï¼»0.20 ± 0.02ï¼½ %, P < 0.01), and so did group G in the body weight (ï¼»88.11 ± 12.65ï¼½ vs ï¼»102.70 ± 16.10ï¼½ g, P < 0.05) and the indexes of the testis (ï¼»0.65 ± 0.13ï¼½ vs ï¼»0.95 ± 0.06ï¼½ %, P < 0.01) and epididymis (ï¼»0.17 ± 0.01ï¼½ vs ï¼»0.19 ± 0.02ï¼½ %, P < 0.05), but no obvious difference was observed in the index of seminal vesicle among different groups. Compared with group A, group B manifested significant decreases in sperm motility (ï¼»74.12 ± 8.73ï¼½ vs ï¼»40.25 ± 6.08ï¼½ %, P < 0.01), and so did group E in sperm count (ï¼»38.59 ± 6.40ï¼½ vs ï¼»18.67 ± 4.59ï¼½ ×105/100 mg, P < 0.01) and sperm motility (ï¼»74.12 ± 8.73ï¼½ vs ï¼»27.58 ± 8.43ï¼½ %, P < 0.01). Sperm motility was significantly lower in group B than in C and D (ï¼»40.25 ± 6.08ï¼½ vs ï¼»58.13 ± 7.62ï¼½ and ï¼»76.04 ± 8.44ï¼½%, P < 0.01), and so were sperm count and motility in group E than in F and G (ï¼»18.67 ± 4.59ï¼½ vs ï¼»25.63 ± 9.66ï¼½ and ï¼»29.92 ± 4.15ï¼½ ×105/100 mg, P < 0.05 and P < 0.01; ï¼»27.58 ± 8.43ï¼½ vs ï¼»36.56 ± 11.08ï¼½ and ï¼»45.05 ± 9.59ï¼½ %, P < 0.05 and P < 0.01). There were no obvious changes in the histomorphology of the testis and epididymis in groups A, B, C and D. Compared with group A, group E showed necrotic and exfoliated spermatogenic cells with unclear layers and disorderly arrangement in the seminiferous tubules and remarkably reduced sperm count with lots of noncellular components in the epididymal cavity, while groups F and G exhibited increased sperm count in the seminiferous tubules and epididymis lumen, also with exfoliation, unclear layers and disorderly arrangement of spermatogenic cells, but significantly better than in group E. CONCLUSIONS: LA can reduce ORN-induced damage to the spermatogenetic function of rats, improve sperm quality, and protect the reproductive system.

Influence of prolonged flaxseed ( Linum usitatissimum ) consumption over epididymis and testicle histoarchitecture of Wistar rats / Influência do consumo prolongado de semente de linhaça ( Linum usitatissimum ) sobre a histoarquitetura dos testículo e epidídimo de ratos Wistar

Flaxseed is considered a functional food with several health benefits. However, because of its high phytoestrogen content, flaxseed influences hormone metabolism and affects the gonadal biomorphology. In this study, computerized histomorphometry was used to evaluate seminiferous and epididymal tubules, considering the different regions of the epididymis (head, body and tail) of rats subjected to a prolonged diet of flaxseed. Young adult male Wistar rats (n=20) were divided into 2 groups during their lactation period: Control Group (CG), fed casein-based meals and Flaxseed Group (FG), fed a 25% flaxseed meal. After 250 days of continuous ingestion, the animals were euthanized and a blood sample was collected. The testicles and epididymis were removed and fixed in buffered formalin solution. The samples were subjected to routine histological paraffin techniques and stained with hematoxilin and eosin. Immunostaining was performed using an antivimentin antibody for Sertoli cell identification. For morphometry, images of the slides were scanned and analyzed using Image J to determine the epithelial height, tubular and luminal diameter and tubular and luminal area. In the hormonal evaluation, FG had a higher serum concentration of estrogen (P=0.001), but no change was observed in the concentration of testosterone. The morphometric assay of seminiferous tubules and epididymal regions revealed no significant differences between the analyzed groups. Similarly, Sertoli cell quantification showed no significant differences in the FG (P=0.98). These results revealed that the continuous and prolonged intake of 25% flaxseed meals from gestation to 250 days of age, even with a significant increase in serum levels of estradiol, does not exert adverse effects on the testicular and epididymal structure or on the cells participating in the spermatogenesis of rats.(AU)

A semente de linhaça é considerada um alimento funcional com vários efeitos benéficos à saúde. Entretanto, devido ao seu elevado teor de fitoestrógenos, esta semente pode influenciar no metabolismo hormonal e interferir na biomorfologia gonadal. Neste estudo, utilizamos a histomorfometria computadorizada para avaliar os túbulos seminíferos e epididimários, considerando as diferentes regiões do epidídimo (cabeça, corpo e cauda) de ratos submetidos a uma dieta prolongada de semente de linhaça. Foram utilizados ratos Wistar machos adultos jovens (n=20) divididos em 2 grupos, durante o período de lactação: Grupo Controle (GC) a base de caseína e Grupo Linhaça (GL) alimentados com 25% de semente de linhaça. Ao final de 250 dias de ingestão contínua, os animais foram sacrificados e amostra de sangue foi coletada. Os testículos e epidídimos foram retirados e fixados em formol tamponado. As amostras foram submetidas ao processamento histológico de rotina para parafina e coradas em hematoxilina e eosina. Foi feita a imunomarcação com anticorpo antivimentina para identificação das células de Sertoli. Para morfometria, as imagens das lâminas foram digitalizadas e analisadas pelo software ImageJ para obtenção dos dados de altura epitelial, diâmetro e área tubular e luminal. Na avaliação hormonal o GL teve maior concentração de estrógeno sérico (p=0,001), mas nenhuma mudança na concentração de testosterona foi observada. Nos parâmetros morfométricos dos túbulos seminíferos e das regiões epididimárias, não houve diferenças significativas entre os grupos analisados. Da mesma forma, a quantificação das células de Sertoli não apresentaram diferenças significativas no GL (p=0,98). Estes resultados mostraram que o consumo contínuo e prolongado de 25% de semente de linhaça desde período gestacional até 250 dias de idade, mesmo com o aumento significativo nos níveis séricos de estradiol, não exerceram efeitos adversos sobre a estrutura testicular e epididimária, assim como nas células participantes da espermatogênese em ratos.(AU)

Protective effects of Urtica dioica L. on experimental testicular ischaemia reperfusion injury in rats.

In this study, it was aimed to examine the effects of Urtica dioica L. (UD) that has antioxidant feature in the experimental testicular I/R model in rats in terms of anti-apoptotic and antioxidative effects. In our study, 24 male rats were divided into three groups: control group, I/R group and I/R + UD (2 mg kg-1 ) group. Seminiferous tubule calibre measurement, Johnson score, haematoxylin-eosin staining, proliferative cell nucleus antigen (PCNA) immunohistochemical staining and TUNEL as histopathological have been conducted. The structural deterioration in the testicular on I/R group has reduced after the treatment of UD. Our data indicate a significant reduction in the activity of in situ identification of apoptosis using terminal dUTP nick end labelling (TUNEL), and there was a rise in the expression of proliferating cell nuclear antigen (PCNA) in testis tissues of UD-treated rats in the I/R group. The I/R + UD group showed a decrease in malondialdehyde levels and an increase in the activities of superoxide dismutase, catalase and glutathione peroxidase in comparison with the I/R group. It could be concluded that protective effects of UD on the I/R testicles are via reduction of histological damage, apoptosis, oxidative stress and lipid peroxidation.

Análise morfométrica dos túbulos seminíferos de camundongos suplementados com óleo de alho / Morphometric analysis of seminiferous tubules of mice supplemented with garlic oil

Introdução: O alho (Allium sativum) é amplamente utilizado na culinária e no tratamento e prevenção de diversas patologias. A literatura carece quanto à toxicidade e os efeitos que o consumo exacerbado de alho pode ocasionar no sistema reprodutor masculino. O óleo de alho é composto por compostos voláteis presentes em seus bulbos. Este estudo tem como objetivo avaliar a morfometria de túbulos seminíferos de camundongos suplementados com óleo de alho. Metodologia: Foram tratados 32 camundongos BALB/c machos divididos em 4 grupos experimentais: 1,4 mg/kg (dose terapêutica); 14 mg/kg; 28 mg/kg; e controle incorporado ao farelo de ração. Após o período experimental os animais foram sacrificados e tiveram os testículos removidos e pesados. Após os procedimentos padrões para preparação das lâminas histológicas, foi medida a espessura dos túbulos seminíferos, através da distância da parede até o lúmen. Resultados e Discussão: Através das medições, não foram observadas diferenças estatísticas que pudessem indicar alterações nestas estruturas em animais suplementados com óleo de alho quando comparados aos animais do grupo controle. A partir deste estudo conclui-se que camundongos suplementados com óleo de alho, em diferentes doses, em um período de 35 dias, não apresentaram alterações na morfometria de túbulos seminíferos que pudessem indicar toxicidade.(AU)

Introduction: Garlic (Allium sativum) is widely used in cooking and in the treatment and prevention of various diseases. The literature lacks for toxicity and the effects that excessive consumption of garlic can cause the male reproductive system. The garlic oil comprises volatile compounds in their bulbs. This study aims to evaluate the morphometric of the seminiferous tubules with garlic oil supplemented mice. Methods: We treated 32 BALB/c mice divided into 4 experimental groups: 1,4 mg/kg (therapeutic dose); 14 mg/kg; 28 mg/kg; and control incorporated into feed meal. After the experimental period, the animals were sacrificed and had their testes removed and weighed. Following standard procedures for preparation of histological slides, we measured the thickness of the seminiferous tubules through the distance from the wall to the lumen. Results and Discussion: Through the measurements of the seminiferous tubules, no statistical differences could indicate changes in these structures in animals supplemented with garlic oil compared with the control group were observed. From this study it is concluded that supplemented mice with garlic oil, in different doses, for a period of 35 days, showed no morphometric changes in the seminiferous tubules which could indicate toxicity.(AU)

The effects of Tynnanthus fasciculatus (Bignoniaceae) infusion on testicular parenchyma of adult Wistar rats.

Traditional medicine provides strong guidance for scientific experiments involving plant products used by the Brazilian people. The species "cipó-cravo" (Tynnanthus fasciculatus) is a plant commonly used either to combat indigestion and stomach aches, or as a general stimulant and aphrodisiac. In this study, the effects of "cipó-cravo" infusion were investigated within the testicular parenchyma of adult Wistar rats. Rats were divided into 3 groups: a control (distilled water) and two treated groups, which received the plant infusion (100 and 200mg/animal/day). The 200mg dose promoted a significant increase of the testicular parenchyma weight and of the volume and total length of the seminiferous tubules, as well as in total daily sperm production and sperm production per gram of testis.

The effects of Tynnanthus fasciculatus (Bignoniaceae) infusion on testicular parenchyma of adult Wistar rats

Traditional medicine provides strong guidance for scientific experiments involving plant products used by the Brazilian people. The species "cipó-cravo" (Tynnanthus fasciculatus) is a plant commonly used either to combat indigestion and stomachaches, or as a general stimulant and aphrodisiac. In this study, the effects of "cipó-cravo" infusion were investigated within the testicular parenchyma of adult Wistar rats. Rats were divided into 3 groups: a control (distilled water) and two treated groups, which received the plant infusion (100 and 200mg/animal/day). The 200mg dose promoted a significant increase of the testicular parenchyma weight and of the volume and total length of the seminiferous tubules, as well as in total daily sperm production and sperm production per gram of testis.

Nitric oxide modulates gonadal and adrenal function in Japanese quail Coturnix coturnix japonica.

Nitric oxide (NO) a gaseous neurotransmitter is reported to play an important role in controlling the release of luteinizing hormone releasing hormone (LHRH) in mammalian system. But its role has not been investigated in birds, where photoperiod plays an important role in regulating gonadal development. To investigate the effect of NO on gonadal and adrenal function of Japanese quail, in the first experiment, 3-weeks-old sexually immature quail received NO donor sodium nitroprusside (SNP, 5mg/100g body weight) orally or nitric oxide synthase (NOS) inhibitor N-nitro L-arginine methyl ester (L-NAME, 25 microg/100g body weight) intraperitoneally for 13 days in continuous condition of light (LL). Thereafter treated quail along with their respective controls were shifted to long day length (LD 16:8) for 21 days when the study was terminated. Results indicate that SNP treatment stimulated and L-NAME suppressed body weight, cloacal gland volume (an androgen dependent sex accessory organ), testes (gonado-somatic index, spermatogenesis), epididymis (histology) and adrenal (weight, histology, cortico-medullary ratio) function as well as total nitrite and nitrate concentration in plasma, hypothalamus and testes. In the second experiment, two groups of 3-weeks-old birds were maintained in short day length (LD 6:18) or long day length (LD 16:8) for 5 weeks to induce gonadal suppression and stimulation, respectively. Thereafter birds of both the photoperiod were divided into two subgroups, the short day quail receiving normal saline (SD Con) or SNP (SD+SNP) while long day quail received normal saline (LD Con) or L-NAME (LD+L-NAME) for 13 days and were maintained in their respective photoperiods, until the termination of study (21 days post treatment period). SNP stimulated all the parameters even in short day condition and L-NAME suppressed in long day quail compared to their respective controls. These findings indicate positive control of NO on the gonad and adrenal function of Japanese quail which exhibits parallel adrenal-gonad relationship. Further, NO donor induces long day effects while NOS inhibitor mimics short day effects. It is concluded that NO may not only regulate hypothalamo-hypophyseal-gonadal and -adrenal axis of Japanese quail but may also modulate its photosexual responses.

Adult-only exposure of male rats to a diet of high phytoestrogen content increases apoptosis of meiotic and post-meiotic germ cells.

Apoptosis plays a critical role in regulating sperm production. Removal of androgens and gonadotropins, or estrogen administration induces germ cell apoptosis. It is hypothesized that dietary phytoestrogens increase apoptosis of developing germ cells, decreasing sperm production. This study aimed to test this in rats fed a high phytoestrogen diet only during adulthood. Male Wistar rats used in this study were offspring of females maintained on a low phytoestrogen diet prior to conception through to weaning. After weaning, juveniles were fed the same low phytoestrogen diet into adulthood. A cohort of males were transferred to a high phytoestrogen diet for 24 days and subsequently testes were collected from all animals. In the high phytoestrogen fed group, homogenization-resistant sperm counts were significantly decreased, as were epididymal sperm counts. Morphometric analysis determined round and elongated spermatid volumes to be significantly decreased, but seminiferous tubule lumen diameters to be significantly increased. TUNEL analysis determined that apoptosis of spermatocytes and round spermatids was significantly greater in the high phytoestrogen fed rats. Neither plasma gonadotropin concentrations nor testicular testosterone were altered. In conclusion, exposure of the adult male rat to a high phytoestrogen diet disrupts spermatogenesis, increasing germ cell apoptosis. This effect is independent of the hypothalamo-pituitary-testicular axis and is likely due to disruption of estrogen's actions in the testis.

Effects of GnRH immunization in sexually mature pony stallions.

Immunization against gonadotrophin releasing hormone (GnRH) was studied as an alternative for the commonly used surgical castration in stallions. Two GnRH vaccines comprising non-mineral oil adjuvants were evaluated for their potential to induce high antibody titers directed against GnRH and subsequent effects on reproductive characteristics. Twelve sexually mature male hemicastrated Shetland ponies were assigned to three groups. Group 1 and 2 were injected with 1mg peptide equivalent of G6k-GnRH-tandem-dimer conjugated to ovalbumin (OVA) in CoVaccine HT adjuvant (GnRH/CoVaccine) and in Carbopol (GnRH/Carbopol), respectively, and group 3 was injected with CoVaccine HT adjuvant without antigen (controls). After immunization no adverse effects were observed with respect to the injections sites or general health. Two weeks after the second vaccination antibody titers against GnRH increased rapidly in all animals of the GnRH/CoVaccine group, at the same time reducing serum testosterone levels maximally for the further duration of the experiment. In the GnRH/Carbopol group antibody responses and effects on testosterone levels were intermediate in two stallions and not apparent in the remaining stallions of this group. Semen evaluation showed that from 2 weeks after the second immunization onwards, sperm motility was affected in all stallions treated with GnRH/CoVaccine and one stallion treated with GnRH/Carbopol. Seven weeks after the second immunization, no semen could be collected from two stallions, one of each group, due to suppressed libido. Histological examination of the testes, 15 weeks after the initial immunization, demonstrated reduction in seminiferous tubuli diameters in all stallions of the GnRH/CoVaccine group and one stallion of the GnRH/Carbopol group. Furthermore, spermatogenesis was extremely disorganized in these stallions, as indicated by absence of the lumen in the seminiferous tubules, the absence of spermatozoa and spermatids in the tubular cross-sections and the impossibility to determine the stage of the tubular cross-sections. Testis size was also substantially reduced in three out of four stallions treated with GnRH/CoVaccine. The results demonstrate that two immunizations with G6k-GnRH-tandem-dimer-OVA conjugate in a suitable adjuvant such as CoVaccine HT caused a rapid and complete reduction of serum testosterone levels in sexually mature stallions, subsequently leading to reduced sperm motility and affected testis function, while no adverse reactions were observed after immunizations.

Effect of Stephania hernandifolia leaf extract on testicular activity in rats.

AIM: The testicular inhibitory effect of the aqueous fraction of methanol extract of Stephania hernandifolia leaf was studied in male Wistar rats. METHODS: The supernatent and the precipitate part of aqueous fractions of the methanol extract of the leaf were gavaged separately to rat at a similar dose of 200 mg/mL per 100 g body weight per day for 28 days. After cessation of treatment, various observations were conducted. RESULTS: In both treated groups, there were significant decreases in the relative weights of the sex organs, the testicular key androgenic enzymes activities, the plasma level of testosterone, the number of different germ cells at stage VII of seminiferous epithelial cell cycle and the seminiferous tubular diameter in comparison to the controls. Neither of the parts had somatic, renal and hepatic toxicity. This study suggested that the active molecules present in the aqueous fraction of methanol extract of Stephania hernandifolia leaves might be steroids as indicated by thin layer chromatography using specific staining substance for steroid molecules. CONCLUSION: In rats, the aqueous fraction of methanol extract of the S. hernandifolia leaves possesses certain testis-inhibitory substances, which may be steroid-like agents.

Testicular sperm extraction by Trucut needle and milking of seminiferous tubules: a technique with high yield and patient acceptability.

OBJECTIVE: To assess the sperm yield and patient acceptability of Trucut needle testicular biopsy followed by seminiferous tubule milking. DESIGN: Prospective case analysis. SETTING: The Regional Fertility Center, Royal Maternity Hospital, Belfast, Northern Ireland, United Kingdom. PATIENT(S): Forty-one males with obstructive azoospermia (normal testicular volume and FSH and LH levels). INTERVENTION(S): Trucut needle testicular biopsies under local anesthetic with milking of the seminiferous tubules. MAIN OUTCOME MEASURE(S): Quantitation of sperm retrieved per biopsy core and patient follow-up by questionnaire. RESULT(S): A mean of 105,634 sperm (range, 5,000-427,800) were retrieved, and the mean biopsy weight was 9.17 mg. Twenty-six subjects found the biopsy painless and 15 were pain-free after biopsy. CONCLUSION(S): The Trucut needle can be used in combination with seminiferous tubule milking to obtain large numbers of sperm in men with obstructive azoospermia.

Action of pineal indoleamines on the reproductive systems of the male C57 mouse and golden hamster.

Intraperitoneal administration of melatonin, methoxytryptamine and methoxytryptophol in the late afternoon to mice exposed to a 14L:10D photoperiod induced appearance of necrotic cells in seminiferious tubules and an increase in incidence of aspermic tubules. The histological appearances of the seminal vesicles and coagulating glands were, however, unaffected by the treatments. Treatment of mice with methoxyindoleacetic acid did not produce discernible changes in the testes and accessory sex organs. Subcutaneous injections of the aforementioned pineal indoles at a dose of 250 micrograms/injection for 3 months to mature male hamsters kept under a 14L:10D photoperiod did not evoke changes in the testes, seminal vesicles and coagulating glands, suggesting a down-regulation of melatonin receptors by administration of a high dose of pineal indole over a prolonged period.