Efficacy of the motile sperm organelle morphology examination (MSOME) in predicting pregnancy after intrauterine insemination

Background: Although the motile sperm organelle morphology examination (MSOME) was developed merely as a selection criterion, its application as a method for classifying sperm morphology may represent an improvement in the evaluation of semen quality. The aim of this study was to determine the prognostic value of normal sperm morphology using MSOME with regard to clinical pregnancy (CP) after intrauterine insemination (IUI).Methods: A total of 156 IUI cycles that were performed in 111 couples were prospectively analysed. Each subject received 75 IU of recombinant FSH every second day from the third day of the cycle. Beginning on the 10th day of the cycle, follicular development was monitored by vaginal ultrasound. When one or two follicles measuring at least 17 mm were observed, recombinant hCG was administered, and IUI was performed 12-14 h and 36-40 h after hCG treatment. Prior to the IUI procedure, sperm samples were analysed by MSOME at 8400x magnification using an inverted microscope that was equipped with DIC/Nomarski differential interference contrast optics. A minimum of 200 motile spermatozoa per semen sample were evaluated, and the percentage of normal spermatozoa in each sample was determined.Results: Pregnancy occurred in 34 IUI cycles (CP rate per cycle: 21.8%, per patient: 30.6%). Based on the MSOME criteria, a significantly higher percentage of normal spermatozoa was found in the group of men in which the IUI cycles resulted in pregnancy (2.6+/-3.1%) compared to the group that did not achieve pregnancy (1.2+/-1.7%; P = 0.019). Logistic regression showed that the percentage of normal cells in the MSOME was a determining factor for the likelihood of clinical pregnancy (OR: 1.28; 95% CI: 1.08 to 1.51; P = 0.003). The ROC curve revealed an area under the curve of 0.63 and an optimum cut-off point of 2% of normal sperm morphology. At this cut-off threshold, using the percentage of normal sperm morphology by MSOME to predict pregnancy was 50% sensitive with a 40% positive predictive value and 79% specificity with an 85% negative predictive value. The efficacy of using the percentage of normal sperm morphology by MSOME in predicting pregnancy was 65%.Conclusions: The present findings support the use of high-magnification microscopy both for selecting spermatozoa and as a routine method for analysing semen before performing IUI.

Effects of Short-term Hyper- and Hypoprolactinaemia on Hormones of the Pituitary, Gonad and -Thyroid Axis and on Semen Quality in Male Beagles

Effects of a short-term hyper- and hypoprolactinaemia on serum concentrations of LH, testosterone and semen quality in six male Beagles were investigated. Blood samples were collected at 3-day intervals for 12 weeks. The time span was divided into five 3-week periods: pre-treatment, metoclopramide (MCP) treatment (0.2 mg/kg orally three times daily), cabergoline (CAB) treatment (5 mu g/kg orally once daily), post-treatment 1 and post-treatment 2. In the latter, only semen characteristics were evaluated. Semen parameters were analyzed once per week during the whole 15-week investigation time. At the end of each period, the effects of a single intravenous injection of thyrotropin-releasing hormone (TRH; 10 mu g/kg) on the secretion of prolactin (PRL), LH, testosterone, thyroid-stimulating hormone and thyroxine (T4) were investigated. Pre-treatment serum PRL concentration increased under MCP (p < 0.05), followed by a decrease under CAB administration (p < 0.05). Luteinizing hormone and testosterone concentrations were not affected. Except for straight-line sperm velocity, semen quality did not differ between collection periods. A single iv TRH injection induced a significant PRL increase at 20 min in all experimental periods except during CAB treatment. Luteinizing hormone and testosterone did not show clear TRH-related changes. Basic T4 levels were significantly reduced after CAB treatment ( p < 0.05). The results of the present study demonstrate that MCP-induced short-term hyperprolactinaemia in male beagles does not seriously affect the hypothalamo-pituitary axis and semen quality.

Effect of Removing Seminal Plasma Using a Sperm Filter on the Viability of Refrigerated Stallion Semen

Cooling of equine semen obtained from some stallions results in lower seminal quality and viability when the seminal plasma (SP) is present. The objective of this study was to evaluate the effect of the removal of SP using a Sperm Filter on the viability of cooled stallion semen. For this purpose, 31 stallions were used. Their ejaculates were divided into three groups: CN, semen was diluted with an extender; FLT, SP was removed by filtration; and CT, SP was removed by centrifugation and cooled to 15°C for 24 hours. Sperm kinetics and plasma membrane integrity were evaluated immediately after collection (T0) and after 24 hours of refrigeration (T1). No difference (P > .05) was noted at T1 for total sperm motility (TM), progressive sperm motility, or plasma membrane integrity when semen samples from all the stallions were analyzed. However, when samples from stallions termed bad coolers were analyzed (TM = <30% at T1), a difference was observed in TM and progressive sperm motility for CN compared with FLT and CT at T1. Sperm recovery was greater when SP was removed using the filter (FLT) to that when the SP was removed by centrifugation (CN) (89% vs. 81%). Thus, we concluded that filtering with a Sperm Filter is an efficient and practical method for removal of SP from stallion ejaculates, with lower sperm loss than centrifugation. We also found that the presence of SP reduces the quality and viability of cooled semen from stallions whose semen is sensitive to the process of refrigeration. © 2013 Elsevier Inc.

Evaluation of Sperm Kinetics and Plasma Membrane Integrity of Frozen Equine Semen in Different Storage Volumes and Freezing Conditions

In the present study, different freezing systems (Styrofoam box and Mini Digitcool ZH 400) and storage volumes (0.5- and 0.25-mL straws) were compared with regard to sperm kinetics and plasma membrane integrity of frozen and thawed semen. For that, three ejaculates from four animals were frozen in Styrofoam box and Mini Digitcool ZH 400 machine. The 0.5-mL straws were thawed at 46°C for 20 seconds, and the 0.25-mL straws were thawed at 46°C for 12 seconds. Statistical analysis was performed using program R of descriptive analysis box plot, followed by analysis of variance using PROC MIXED of SAS 9.1 package. Variances of 5% were considered as different. There was no interaction between the straw sizes and volumes; however, statistical differences were observed between the semen storage volumes. The 0.5-mL straws had higher total motility (%), progressive motility (%), average path velocity (μm/s), straight-line velocity (μm/s), curvilinear velocity (μm/s), and rapid sperm percentage (%) than the 0.25-mL straws. However, plasma membrane integrity analysis did not differ between the two straws. Thus, it is possible to conclude that equine sperm cryopreserved in 0.5-mL straws has better sperm kinetics than when stored in 0.25-mL straws. Additionally, it is possible to conclude that automated systems that enable faster freezing rates result in a seminal quality that is similar to the one obtained by the conventional system using Styrofoam boxes. © 2013 Elsevier Inc.

Efeito da idade do homem na avaliação do sêmen pela Motile Sperm Organelle Morphology Examination (MSOME)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Eficácia do motile sperm organelle morphology examination (MSOME) na predição de gravidez após inseminação intrauterina

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Exploratory analysis of differences in sperm morphology in Nelore and Gir (Bos indicus) bulls

The aim of this study was to evaluate the effects of breed and season on semen quality parameters of zebu bulls. Data (1,632 registers) of semen production from Gir (n = 4) and Nelore (n = 15) bulls were collected between October 2005 and November 2009. The ejaculates were collected twice a week during various seasons (summer, fall, winter, and spring) and evaluated for the following semen parameters: ejaculate volume, sperm concentration, sperm motility, forward progressive motility (FPM), and sperm morphology. Factor analysis was used to determine the relationship among variables. The effect of breed (Gir and Nelore) and season and their cross effect on each parameter and extracted factor were tested using ANOVA. A negative correlation (P < 0.05) was observed between FPM and proximal droplet, as well as with abnormal loose head, abnormal small head, pouch formation, abnormal mid-peace, and strongly folded tail. Gir bull sperm showed more major defects, detached acrosome, and minor FPM (P < 0.01), whereas Nelore bulls showed a higher number of sperm with normally loose head.

The Influence of Dietary Lysine on Yellow Perch Female Reproductive Performance and the Quality of Eggs

The objectives of this study were to (1) evaluate if dietary lysine (Lys) has an effect on the free amino acid (FAA) pool of Yellow Perch Perca flavescens eggs, and (2) determine how dietary Lys influences the reproductive performance and eggs fertilization rate (embryo survival) of female Yellow Perch. Two-year-old Yellow Perch of initial size of approximately 75 g were randomly distributed into six 400-L tanks at 32 +/- 1 fish per tank. This experiment included two wheat gluten-based diets in triplicate Lys-deficient ([-] Lys) and Lys-supplemented ([+]Lys; 2.23% in dry feed) diets. Females from the reference group were fed a commercial diet. Females from reference, (+) Lys, and (-) Lys groups were stripped and their eggs divided into 0.4-1.2-g portions and mixed with sperm (21.4 +/- 4.3 mu L) from either reference, (+) Lys, or (-) Lys males. The mean weight of Yellow Perch females and mean total weight of ovulated eggs were the greatest in the reference group compared with both (+) Lys and (-) Lys groups. There were no differences in the ratio of weight of eggs to female body weight as well as egg size among groups. There was no difference among treatments in the concentration of free amino acids except glutamic and aspartic acids in Yellow Perch eggs. There was significant effect of female dietary treatments on the egg fertilization rate averaged across all males. The higher fertilization rate was observed in the reference and (+) Lys groups compared with the (-) Lys group. The effect of female dietary treatment on the egg survival was also dependent on the dietary treatment of males.

Improvement of gamete quality and its short-term storage: an approach for biotechnology in laboratory fish

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