Spermatogenesis and karyotypes of three species of water striders (Gerridae, Heteroptera).

Although they are of economic importance, there have been few cytogenetic studies of the Gerridae (Heteroptera) in Brazil. We examined spermatogenesis (meiosis and spermiogenesis) and nucleolar behavior in three species of the family Gerridae. Brachymetra albinerva and Halobatopsis platensis were found to have a chromosome complement of 2n = 25 (24A + X0) and Cylindrostethus palmaris 2n = 29 (28A + X0) chromosomes. Fifteen individuals of these species were collected from the reservoir of São José do Rio Preto, SP, using screens and were transported in pots containing water to the laboratory, where cytogenetic preparations were made. The polyploidy nuclei are formed by several heteropyknotic regions; cells in meiotic prophase have a heteropyknotic region that is probably the sex chromosome, and the chromosomes from chiasmata. The spermatids are rounded and have a heteropyknotic region at the periphery of the nucleus; the sperm head is small, with a long tail. Silver impregnation of meiotic cells showed one or more disorganized bodies around the perichromosomal sheath. The round spermatids had two bodies next to each other, but these were elongated; one of the bodies remained in the head and the other migrated to the initial part of the tail at the end of spermagenesis, when the staining was no longer evident. The meiotic cells appear during spermatogenesis and have very similar silver-impregnation patterns in different species of Heteroptera.

Comparative study of spermatogenesis and nucleolar behavior in testicular lobes of Euschistus heros (Heteroptera: Pentatomidae)

In some testicular lobes of the Pentatomidae there may be occurrence of atypical spermatogenesis or polymegaly, leading to the production of nonfertile sperm. The comparative analysis of spermatogenesis and nucleolar behavior in testicular lobes of Euschistus heros showed cells with polymegaly in lobes 4 and 6. Generally, when these lobes are present in the same individual, there is also the formation of atypical cells in the flanking lobe. Such characteristic was not seen in E. heros. However, differences regarding the concentration of heteropyknotic chromatin and silver-positive bodies in this lobe deserve attention. This study explored the literature and demonstrated the prevalence of some lobes in the formation of differentiated cells. It was also found in the literature that there is an association of the chromocenter with the nucleolus in several species of Pentatomidae, but in E. heros this association does not appear to occur. © 2010 Hederson Vinicius de Souza and Mary Massumi Itoyama.

Reproductive technology in domestic carnivorous

Background: The delay in development of artificial reproduction techniques on carnivorous could be due to countless reasons, but the lack of commercial interest is probably the most important one. The majority of canines are small structures, canidae are extremely fertile and a great number of species are adapted to domestication or captivity. Finally, the canine gamete physiology presents a difficult adaptation of technology knowledge obtained from other species. Furthermore, domestic felines are animals of company and there is no interest in reproducing them in a large scale, as it has been observed in other domestic animals, however, besides of being a valuable model for the development of in vitro techniques, the domestic cat is also used as an embryo receptor for different species of small wild felines due to physiological similarities among them, in vitro embrionary development, Review: It was reviewed the main insights about the reproductive physiology in female dogs, in vitro oocytary maturation (IVM), pregnancy and conception rate with dogs' frozen/unfrozen semen and PIV in domestic cats. The majority of mammal oocytes restart meiosis spontaneously after ovulation and reaches MII in artificial environment; in an in vitro maturation system in bovines, around 90% of oocytes complete their maturation, although its development capacity can be reduced subsequently. The success of IVM in canidae have been limited, with maturation rate varying from 0 to 58%, usually around 20%. The greatest difficulties include oocyte quality, hormonal environment, protein supplementation, cumulus / oocyte cell interaction, donor breed and age, culture systems, oxygen tension, amino acids, growth factor and sequential means. The freezing process reduces the quality of the semen, firstly because it reduces the number of living sperms and secondly because freezing produces cell modifications that could alter the sperm motility, longevity, integrity of membranes and its fertilizing capacity. Conclusion: Nowadays, several researches are being performed with the aim of increasing viability after dogs' and cats' semen is unfrozen, using extenders, cryoprotectors, freezing and unfreezing curves, addition of antioxidant substances. The aim of this text is to inform about the improvements obtained on the artificial reproduction techniques, emphasizing the oocytary maturation in female dogs, semen cryopreservation and artificial insemination in domestic dogs and cats.

Perinatal exposure to parasiticide ivermectin: Long-term effects on reproductive parameters of adult male rats

Ivermectin is one of the most widely used antiparasitic drugs globally. The aim of this study was to evaluate the chronic effects of perinatal exposure to ivermectin on male reproductive parameters in rats. Pregnant rats were treated daily by oral gavage with 0.4 or 1.6 mg kg -1 of ivermectin or vehicle, from gestational day 6 until post-natal day 10. In the adulthood stage, there were significant reductions in the relative testicular weight of rats exposed to the low dose and in relative prostate weight of male rats exposed to the high dose of ivermectin. Furthermore, the animals exposed to the low dose also presented an increased seminal vesicle weight compared to controls. However, neither of the ivermectin doses interfered in daily sperm production, sperm number in testis, or sexual behavior of exposed males. In conclusion, perinatal exposure to ivermectin neither altered the male reproductive system development markedly, nor produced any adverse effects on the parameters evaluated. © 2011 Taylor & Francis.

Relevância do assited hatching com laser em um programa de ovo-doação utilizando banco de óvulos criopreservados: Um estudo prospectivo randomizado

Objective: This prospective randomized trial evaluated if there is an improvement in clinical outcomes when assisted hatching is performed in embryos derived from vitrified oocytes in an ovum donation program. Methods: Sixty oocyte recipients undergoing donation program using egg-cryobanking were randomly allocated to the assisted hatched (AH, n=30) or control group (n=30). Pregnancy and implantation rates were compared between the groups. Vitrification and warming procedure were carried out according to the Cryotopmethod. Immediately before embryo transfer, embryos undergoing laser-assisted hatching had the zona pellucida thinned using a 1.48 μm wavelength diode laser. Results: A total of 288 vitrified MII oocytes were warmed for the 60 recipients (4.8 oocytes per recipient). Out of 288 vitrified oocytes, 273 (94.8%) survived. All surviving oocytes were sperm injected and 228 displayed 2 pronucleus 16-18h after injection (83.5%). There were 172 good quality embryos transferred. Twenty four patients achieved clinical pregnancy (total pregnancy rate of 40%). The clinical pregnancy rate did not differ between AH and control groups (44.4% and 33.3%, respectively, p=0.1967), however AH resulted in a significant higher implantation rate (31.6% and 18.4%, p=0.0206). These findings were confirmed by the regression models either for pregnancy (OR = 1.14; IC 95% = 0.80-.72; p= 0.766), as for the implantation rate (RC:19.45, P=0.041). Conclusions: Our evidences demonstrated the effectiveness of the AH in embryos derived from warmed oocytes and suggest that oocyte cryopreservation is a valuable tool to provide successful outcomes in an egg donor program. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Locus no cromossomo 19 influencia positivamente o número de oócitos recuperados em ciclos estimulados

Objective: To evaluate if identified loci associated with normal age of menopause variation and early menopause can account for the poor response to controlled ovarian stimulation. Methods: A total of 71 patients, with age ≤ 35 years old, undergoing intracytoplasmic sperm injection were tested for three of the four newly identified genetic variants associated with normal variation in menopausal age and early menopause. Patients were divided into two groups: poor responder group (PR group, n=21) and normoresponder group (NR group, n=50). The influence of risk allele frequency on the response to controlled ovarian stimulation (COS) was evaluated. Logistic regression models were used. Results: There was no significant difference in the incidence of the genetic variants between NR and PR group. The risk allele for chromosome 19 variant (rs4806660) demonstrated a protective effect. The presence of a risk allele, either in homozygosis or in heterozygosis, was associated with an increased response to COS, resulting in an elevated number of follicles (Coef: 2.54, P= 0.041) and retrieved oocytes (Coef: 1.41, P= 0.041). Conclusions: Genetic variants rs244715, rs9379896 and rs4806660 are not risk factors for poor ovarian response. Instead, rs4806660 is associated with higher number of follicles and retrieved oocytes. It could be hypothesized that rs4806660 is associated with an increased response to gonadotrophin stimulus. © Todos os direitos reservados a SBRA - Sociedade Brasileira de Reprodução Assistida.

Use of bayesian inference to correlate in vitro embryo production and in vivo fertility in Zebu Bulls

The objective of this experiment was to test in vitro embryo production (IVP) as a tool to estimate fertility performance in zebu bulls using Bayesian inference statistics. Oocytes were matured and fertilized in vitro using sperm cells from three different Zebu bulls (V, T, and G). The three bulls presented similar results with regard to pronuclear formation and blastocyst formation rates. However, the cleavage rates were different between bulls. The estimated conception rates based on combined data of cleavage and blastocyst formation were very similar to the true conception rates observed for the same bulls after a fixed-time artificial insemination program. Moreover, even when we used cleavage rate data only or blastocyst formation data only, the estimated conception rates were still close to the true conception rates. We conclude that Bayesian inference is an effective statistical procedure to estimate in vivo bull fertility using data from IVP. © 2011 Mateus José Sudano et al.

Análise espermática de perdizes (Rhynchotus rufescens) criadas em cativeiro e suplementadas com selênio

Due to the commercial importance of the red-winged tinamou (Rhynchotus rufescens), for the past few years, the employment of reproductive biotechnologies has been attempted. Thirty animals were randomly assigned into two groups: control group (no selenium) and treatment group (supplemented with 0,2 a 0,8 mg selenium/ 1000 kg ration). Animals were allocated at the FCAV - UNESP/Jaboticabal (2007-2008). Semen collections were performed by digital manipulation and divided in pools of at least 150 μL. After the immediate evaluation of motility, vigour, concentration and morphology, an aliquot of 20 μL was diluted in 300 μL of physiologic solution in order to test acrosome and membrane integrities, which were performed by counting 200 cells for each test. Cells were evaluated as follows: 1) Intact acrosome: lilac acrosome; Non-intact acrosome: pink acrosome; 2) Live cells: non stained; Dead: stained. Data was statistically analysed using the SAS System for Windows. No differences were found between treatment and control groups for volume, motility, vigour, mean number of spermatozoa per animal, concentration, Intact acrosome, Intact membrane. The difference found on midpiece sperm defect (Se = 1,33 ± 0,53 and control = 3,78 ± 0,69, p = 0.0107) may be due to the damages caused by the selenium deficiency to the architecture of the midpiece, which compromises sperm mobility and fertilization capacity.

The expression of aquaporins 1 and 9 in adult rat epididymis is perturbed by chronic exposure to ethanol

Aquaporins (AQPs), notably AQP-1 and AQP-9, may contribute to reabsorption of fluid and solute across the epididymis. Ethanol is related to be a toxicant affecting directly or indirectly the epididymis and the sperm motility. This study examined the expression of AQP-1 and AQP-9 in adult epididymis of the UChA and UChB 10% (v/v) ethanol-preferring rats, focusing the ethanol-induced hormonal disturbances upon the regulation of these AQPs. Chronic ethanol intake significantly decreased body weight, while UChA and UChB rats displayed a marked loss of epididymal weights. Both ethanol-consuming animals had a severe reduction of testosterone levels, whereas LH and 17β-estradiol were unchanged. Throughout the epididymis, a strong reaction to AQP-1 was observed in myoid and endothelial cells of the UChB ethanol-preferring rats, differently from a moderate intensity in the initial segment of the UChA rats. In addition, AQP-9 showed a strong immunoreaction in the apical membrane of principal cells at initial segment. In cauda epididymis, the level of AQP-9 was reduced along the microvillus projections in both UChA and UChB rats compared to controls. We conclude that chronic ethanol consumption modulates the androgen levels, thereby modifying the expression pattern of AQP-1 and 9 in the epididymis. © 2011 Elsevier Ltd.

Use of cooled bull semen as a strategy to increase the pregnancy rate in fixed-time artificial insemination programs-case report

Semen cryopreservation is still considered suboptimal due to lower fertility when compared to fresh semen. The reasons for the loss of fertility are various and related to irreversible damage caused to the cells during the freeze-thaw process. An alternative to conventional cryopreservation represents the use of chilled bull semen, preventing the damage associated with freezing, thereby guaranteeing greater sperm viability. The aim of this study was to describe the use of cooled bull semen as a strategy to increase the pregnancy for Fixed-Time Artificial Insemination (FTAI) of Nellore (Bos indicus) cows. One ejaculate of a select Nellore bull obtained by electroejaculation was used; the semen sample was fractioned into two aliquots: one diluted in Botu-Bov® extender containing 6.4% glycerol for cryopreservation (BB-F, frozen group) and one diluted in the same extender, free from cryoprotectants and used for cooling (BB-C, cooled semen group). The samples in the BB-C group were chilled to 5°C using an isothermic box and maintained for 24 h prior to use. A total of 349 lactating Nellore cows (70-90 days after birth) were synchronized by the insertion of a progesterone releasing device (1.0 g) and estradiol benzoate (2.0 mg i.m.) on a random day of the estrous cycle (Day 0); FTAI was performed 44-48 h after the removal of the device. The pregnancy rates were 45.71 and 61.49% (P<0.05), respectively, for the cryopreserved or chilled bovine semen groups. In conclusion, the use of bull semen cooled for 24 h represents an alternative to conventionally cryopreserved semen, as determined by the increase the pregnancy per artificial insemination in bovine herds. © 2012 Science Publication.

Idade masculina: Qualidade do sêmen e fertilidade: Idade masculina X qualidade seminal

The man's age seems to affect reproductive function but to a lower degree compared with the woman's age. In fact, the effect of paternal age on semen quality has been discussed, but the results are contradictory. Considering the differences in the results of studies, this review aimed to analyze the literature regarding the effect of paternal age on semen parameters (volume, concentration, motility and morphology) in sperm DnA damage in assisted reproduction results and genetic diseases.

Congelação de espermatozoides epididimários de gatos utilizando o diluidor Botu-crio® após refrigeração por 24 h em contêiner de transporte de sêmen Botu-tainer®

The possibilities of using the sperm collected from the epididymis have been widely used because the fertilizing capacity sperm preservation and the possibility of using it for wild cats. But in the process of cryopreservation, some studies show a decrease in the quality of the sperm when left under cooling before frozen for some time. This study aimed to assess the quality of the epididymal sperm obtained from domestic cats after cryopreservation using a diluent based on egg yolk and glycerol (Botu-crio®), comparing the morphofunctional characteristics after cooling for 24 hours in a container of semen transport (Botu-tainer®). We use eight cats submitted to elective orchiectomy, aging from eight months, without racial determination, and good nutritional status. These sperm characteristics were: motility, vigor, concentration, membrane integrity and morphology. It has been found, after statistical analysis, that the container of semen was able to maintain sperm viability, even for 24h. We also observed a significant decrease on all parameters after frozen, consequential, probably to thermal stress that occurs in processing. However, the percentage of membrane integrity after thawing shows good employability of the Botu-crio®, which viability is possible to perform in vitro fertilization, requiring higher ratings.

Possible mechanism by which zinc protects the testicular function of rats exposed to cigarette smoke

Background: The aim of this study was to evaluate the changes in testicular function of rats due to cigarette smoke exposure and the possible mechanism by which zinc protects against these alterations. Methods: MaleWistar rats (60 days old) were randomly divided into 3 groups: control (G1, n = 10); exposed to cigarette smoke (G2, n = 10; 20 cigarettes/day/9 weeks) and exposed to cigarette smoke and supplemented with zinc (G3, n = 8; 20 cigarettes/day/9 weeks; 20 mg/kg zinc chloride daily for 9 weeks, by gavage). After the treatment period, the animals were euthanized, and materials were collected for analyses. Results: G2 rats showed a reduction in body mass; impaired sperm concentration, motility, morphology and vitality; and increased malonaldehyde and thiol group levels and superoxide dismutase activity as compared to G1. Zinc prevented the reduction of sperm concentration and the excessive increase of lipid peroxidation and induced an increase in plasma testosterone levels, wet weight of testis and thiol group concentration. Conclusions: Exposure to cigarette smoke led to harmful effects on testicular function at least partially due to the exacerbation of oxidative stress. Supplementary zinc had an important modulator/protector effect on certain parameters. The mechanism of zinc protection can be through an increase of SH concentration. Thus, zinc supplementation may be a promising addition to conventional treatments for male infertility related to smoking. Copyright © 2012 by Institute of Pharmacology Polish Academy of Sciences.

Morfologia do epidídimo de cobaio (Cavia porcellus) em diferentes fases do desenvolvimento pós-natal

The morphological evolution of the epididymal duct of guinea pigs (Cavia porcellus, L.) was studied on 10, 20, 30, 45, 60, 70, 90 and 100 days of age, being complex, which is due to the proper differentiation postnatal in the epididymal epithelium. Thus, it was observed that the initial segment of the epididymis reveals an increase of epithelial height corresponding to the average height of the main tubular epithelium cells, generally progressive after 45 days of age. The epithelial height in the middle segment were higher in younger stages (10 to 45 days) than in the prepubertal age (60 days), and have a decrease among 70 to 100 days of age. The ductular terminal segment starts a gradual decrease of the epithelial heights from puberal age (70 days), until adult age (90 to 100 days). In addition, this segment showed epithelial waviness that disappeared after 70 days of age, when the lumen fills it with sperm and cellular exfoliation. The cell types of the epididymal tubular epithelium, principal cells, basal cells and apical cells, were observed in all ages. In the epididymis of very young animals, there was a predominance of undifferentiated columnar cells. After 20 days of age, there was natural prevalence of the principal cells on other cell types, what is a feature similar to other mammals.

Morphological Variation of Primary Reproductive Structures in Males of Five Families of Neotropical Bats

Bats present unique features among mammals with respect to reproduction, and although neotropical bats do not have a hibernation period, many of their reproductive characteristics vary seasonally and interspecifically. Thus, this work aimed to examine the reproductive structures of 18 species belonging to five families of Brazilian bats. The gross anatomy of the testes varied little; however, the epididymis of Emballonuridae and Vespertilionidae showed exceptional structures with a large elongation of the caudal region. We observed a wide variation in the positioning of the testes: Phyllostomidae and Noctilionidae presented external testes; Emballonuridae and Molossidae presented migratory testes that may be in intra-abdominal or external positions; and Vespertilionidae displayed scrotal testes. In the histological evaluation, we observed a different pattern in vespertilionid species, with testicular regression and sperm retention/storage in the cauda epididymis in the five species analyzed. Similar testicular regression was observed in Molossops temminckii; however, sperm retention/storage was not observed in this species. These data suggest that although the species analyzed are tropical species that do not present a prolonged period of torpor (hibernation), they still maintain a period of seminiferous tubule regression and sperm storage very similar to that observed in hibernating bats. © 2012 Wiley Periodicals, Inc.