Short- and long-term reproductive effects of prenatal and lactational growth restriction caused by maternal diabetes in male rats

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Decreased Sperm Motility in Rats Orally Exposed to Single or Mixed Pesticides

The Brazilian Agency of Sanitary Vigilance (ANVISA) conducted a study that demonstrated the presence of residues of several pesticides in fresh fruits and vegetables that were available for purchase by the general populace. In order to evaluate potential adverse health effects of low-level exposure to agrochemicals, the reproductive toxicity of the pesticides dicofol, dichlorvos, permethrin, endosulfan, and dieldrin was evaluated in rats dosed with these chemicals individually or as mixtures. Sixty male Lewis rats (6 wk old, 200 x g) were randomly allocated to 8 groups: (1) control group, received basal diet; (2) 5 groups designated a to e received the diet containing each pesticide individually, at the respective effective doses: lowest-observed-adverse-effect level (LOAEL) for dieldrin and endosulfan, lowest-observed-effect level (LOEL) for dicofol, and lowest effect level (LEL) for dichlorvos and permethrin, respectively, depending on the published data; (3) effective dose group, which received a mixture of pesticides added to basal diet at the respective doses reported to produce adverse effects; and (4) low dose group, which received a pesticide mixture added to the basal diet, where each pesticide was at its no-observed-effect level (NOEL). After 8 wk of treatment, reproductive parameters were evaluated. Sperm morphology, daily sperm production (DSP), sperm transit time through the epididymis, hormonal levels, and histopathological evaluation of testis and epididymis did not differ significantly among the groups. However, sperm motility was significantly decreased in animals that received a mixture of dieldrin, endosulfan, dicofol, dichlorvos, and permethrin, as well as in the group receiving dicofol alone. Exposure to the individual pesticides endosulfan, dichlorvos, and permethrin did not markedly affect sperm motility. The impairment of sperm motility in the mixture of pesticides at the NOEL level indicates that reproductive effects not seen with individual pesticides may occur in presence of several pesticides due to an additive effect. However, the pesticide mixtures did not appear to affect DSP or spermatogenesis despite reduced sperm motility.

Morphological and functional characteristics of chilled semen obtained from domestic feline epididymides (Felis catus)

The objective of this study was to determine morphological and functional characteristics of semen retrieved from the feline epididymis before and after cooling. Sixteen adult male cats were orchiectomized. The distal portion of the epididymis and proximal part of the deferent ducts were dissected and squeezed to obtain their content. After centrifugation, the supernatant was removed, sperm were resuspended in a 0.9 mL Tris-fructose-citric acid extender containing 20% egg yolk, aliquoted into three 0.3 mL samples, placed in a refrigerator (4.8 degrees C) and cooled (0.5 degrees C/min). Semen evaluations were performed on four occassions: immediately after epididymal sperm retrieval (TO), and at 24 h (T-1), 48 h (T-2) and 72 h (T-3) after cooling. on each occasion, progressive motility, vigor and sperm morphology were determined. Mean motility and vigor decreased (P < 0.05) between each successive examination. Although the majority of sperm cell damage occurred within the first 24 h, there was a decrease (P < 0.05) in mean percentage of morphologically normal sperm between To and each evaluated time (T-1, T-2, T-3) after cooling, due to an increase in coiled and bent sperm tails. Further studies are needed to evaluate the effects of cooling on the fertilizing capacity of cat epididymal spermatozoa in assisted reproduction programs. (c) 2006 Elsevier B.V. All rights reserved.

Evaluation testicular fine needle aspiration cytology and serum testosterone levels in dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cryopreservation and fertility of ejaculated and epididymal stallion sperm

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Advances in stallion's epididymal sperm technology

Freezing epididymal sperm is a method to preserve germplasm from animals with not only high genetic potential but also endangered species. In the equine some owners have chosen this possibility in cases of either severe illness or death of stallions. However, the lack of knowledge and poor published results of such technique hampers its propagation. New procedures have allowed some improvement on fertility rates of frozen sperm from the epididymis of stallions. The aim of this study is to report the advances on processing and cryopreservation of samples from the stallion's epididymal semen.

Histopathology of the reproductive system of male sheep experimentally infected with Toxoplasma gondii

The aim of this study was to investigate the histopathological changes in reproductive system (testicles, epididymis, seminal vesicles, and prostate) of small male ruminants after Toxoplasma gondii infection. Eight sheep were inoculated with T. gondii: group I, four sheep (2.0 x 10(5) P-strain oocysts); group II, four sheep (1.0 x 10(6) RH-strain tachyzoites); and group III, two uninfected sheep maintained as control. Infection with T. gondii was confirmed by seroconversion (indirect fluorescent antibody test-IgG) in all the infected animals beginning on post-inoculation day (PID) 7. on PID 70, all the animals were euthanized and tissue samples (testicles, epididymis, seminal vesicles, and prostate) were collected and processed for histological analysis. The main changes detected were a focal mononuclear interstitial inflammatory infiltrate in the prostate and seminal vesicles; diffuse testicular degeneration associated with calcification foci and a multifocal mononuclear interstitial inflammatory infiltrate; and a mononuclear interstitial infiltrate and focal necrotic areas of the muscle fibers surrounding the seminal vesicles. The histopathological findings of this work, along with the detection of T. gondii in the examined parenchyma tissues (immunohistochemistry) and the results obtained by other authors examining different tissues, suggest that histological changes diagnosed in the reproductive system of rams infected with T. gondii are strongly suggestive of toxoplasmatic infection.

Morphological features of the epididymal epithelium of gerbil, Meriones unguiculatus

Principal cells of the ducts epididymis of the Mongolian gerbil showed ultrastructural characteristics of lining epithelium cells close related to processes of protein secretion, and transcytosis occurring between adjacent principal cells which were mainly verified in the initial segment. Principal cells also presented roles of fluid phase and adsorptive endocytoses, as well as autophagic and heterophagic lysosomal activities mainly observed in the caput epididymis. Columnar (principal) cells of the corpus epididymidis presented great number of variable vesicles and vacuoles distributed in all the cytoplasmic levels occurring a progressive coalescence pattern among them, which help to guarantee formation of cytoplasmic channels for fluid phase transport between the tubular lumen and epididymal interstitium. Clear cells were presented in the initial segment and predominately in the cauda epididymis epithelium of the gerbil and showed marked ultrastructural characteristics of endocytosis activities occurrence, perhaps directly related to the turnover of fluid phase of spermatozoa stored into the lumen of the distal tail. Other epididymal epithelium cells were verified and described such as basal, halo, apical and dark cells, but they did not presented special ultrastructural features. (c) 2007 Elsevier Ltd. All rights reserved.

Aquaporin 9 (AQP9) localization in the adult dog testis excurrent ducts by immunohistochemistry

Aquaporins (AQPs) are small, intrinsic membrane proteins that are present in many cell types involved in fluid transport. AQP9 is a major apical water channel that is expressed throughout the efferent ducts, epididymis, and vas deferens, as well as in other regions of the human and rodent male reproductive tract. The target of this study was to examine the expression of AQP9 in epithelial cells in the adult dog efferent ducts, epididymis, and vas deferens. Samples of dog male reproductive tract comprising fragments of the testis; initial segment, caput, corpus, and cauda of the epididymis; and vas deferens were obtained from eight adult mongrel dogs. Immunohistochemistry and Western blotting procedures were used to show AQP9 localization and distribution. AQP9 expression was not detected either in dog seminiferous tubules or rete testis. However, apical labeling for AQP9 was detected in the different regions of epididymis and vas deferens, with the reaction being less intense in the caput epididymis. Thus, AQP9 is abundantly expressed in dog male reproductive tract, in which it is an important apical pathway for transmembrane flow of water and neutral solutes.

Ultrastructure of the epithelium lining of cauda epididymidis in mongrel dogs

The epithelium lining of cauda epididymidis in mongrel dogs was examined by transmission electron microscopy. The epididymal epithelium is pseudostratified with stereocilia and is composed predominantly of principal and clear cells. Therefore, exist basal and apical cells. The principal and clear cells show features suggesting that they may be preferentially involved in absorptive and secretive functions. These results are compared with previously published data on the cauda epididymidis of other mammalian species, in order to understand the significance of the epididymis in sperm maturation.

Neuroendocrine and reproductive aspects of adult male rats exposed neonatally to an antiestrogen

The present study was designed to investigate the effects of a single dose of an estrogen antagonist-clomiphene-during neonatal life, on later neuroendocrine system and reproductive performance. Immediately after birth, male pups received clomiphene citrate (s.c.). At adulthood, although testosterone levels and wet weights of reproductive organs were not altered, the treatment induced an increased number of spermatozoa and a delay in the transit time in the cauda epididymis. Additionally, there was impairment of sexual behavior evidenced by a delay in the latencies to the first mount and first intromission. Treated rats also showed decreased dopaminergic and serotonergic neurotransmissions in the hypothalamus and decreased dopaminergic neurotransmission in the striatum. The decreased dopaminergic activity could be related to the lower sexual motivation observed. These results indicate the necessity of preventing exposure to drugs that may impair sexual differentiation, which can compromise later mating success as well as the capacity to generate descendants. (c) 2006 Elsevier B.V. All rights reserved.

Impairment of adult male reproductive function in rats exposed to ethanol since puberty

The objective of this work was to evaluate reproductive function in adult male rats exposed to ethanol since puberty. Male Wistar rats, 50 days old, received a liquid diet with 36% of the daily calories derived from ethanol or an isocaloric control diet for 55 days. The ethanol treatment impaired sexual behavior and only 22% of these rats reached ejaculation. The fertility of ethanol-treated animals was significantly reduced, mainly after natural mating. Serum testosterone levels, daily sperm production and sperm count in the epididymis were also significantly diminished after ethanol treatment, associated with an acceleration of the sperm transit time in the cauda epididymidis, decrease in sperm motility and increased percentage of abnormal shaped sperm cells. The results showed that chronic consumption of ethanol beginning at puberty impairs the reproductive function of adult male rats. (c) 2006 Elsevier B.V. All rights reserved.

Effects of prenatal hydrocortisone acetate exposure on fertility and sexual behavior in male rats

The aim of the present study was to investigate the effects of hydrocortisone during the prenatal period and its later repercussions on the fertility and sexual behavior of male rats. Pregnant rats were treated (s.c.) with hydrocortisone acetate, at 1.5 mg/day on the 17th, 18th, and 19th days of gestation. Decreased body weight and no alteration in anogenital distance were observed in male offspring. Adulthood, presented reductions of body weight, plasma testosterone levels, and seminal-vesicle wet weight without secretion as well as no alteration in the wet weights of the testes, epididymis, and seminal vesicle with secretion in the treated group. Males exposed to hydrocortisone during the prenatal period were able to mate with normal females, which became pregnant but exhibited an increased number of post-implantation losses. In spite of this, these treated males exhibited decreased male sexual behavior and the appearance of female sexual behavior after these male rats were castrated and pretreated with exogenous estrogen. These results indicate that exposure to hydrocortisone in the later stages of pregnancy may have a long-term effect on the fertility and sexual behavior of mate rats, suggesting an incomplete masculinization and defeminization of the central nervous system. (C) 2003 Elsevier B.V. (USA). All rights reserved.

Decrease in sperm number after treatment of rats with Austroplenckia populnea

The purpose of this study was to evaluate the effects of a hexanic extract (HE) made from leaves of A. populnea collected in Botucatu, State of São Paulo, and Nova Lima, State of Minas Gerais, Brazil, at a range of doses during 7 and 14 days, on the male reproductive system of rats. The treatment did not affect the body weight, nor absolute organ weight. The serum testosterone levels, testicular sperm head counts, daily sperm production, and sperm morphology did not differ from that of the control groups. The spermatogenesis and the morphometric parameters of cauda epididymidis were not affected by the treatment. Cauda epididymis sperm number was significantly reduced in the group that received HE of Nova Lima, 1 g/kg/day, during 14 days, from the control group. (C) 2000 Elsevier B.V. All rights reserved.

Fenvalerate, A Pyrethroid Insecticide, Adversely Affects Sperm Production and Storage in Male Rats

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)