Determine Semen Quality Research Articles

Assessment of Sperm Morphometry in Evaluating Male Infertility

Background: Infertility is a complex issue affecting 15% of couples of reproductive age, with men accounting for 40%-50% of infertility cases. Semen analysis comprises various descriptive measures of sperm and seminal fluid to determine semen quality. Transforming qualitative descriptions of sperm deformities and shape changes into quantitative terms can aid in identifying sub-visual abnormalities. This study aimed to evaluate sperm morphometry parameters in both infertile and fertile men. Methods and Results: The study enrolled a total of 101 participants, divided into three groups: Group A included 38 subfertile patients with varicocele, Group B included 33 patients with idiopathic infertility (23 with asthenozoospermia and 10 with oligozoospermia), and Group C (the control group) included 30 healthy fertile men. The mean age of patients was 31.6±5.81, 31.3±6.0, and 29.47±4.27 years in Groups A, B, and C, respectively. Scrotal duplex examinations were performed to identify the presence of varicocele. Semen samples were collected following WHO Manual (2010). Semen dynamic and morphological analyses were conducted using CASA (Computer-Assisted Semen Analysis, MIRALAB, ISO9001, ISO13485). We found that sperm concentration, total sperm count, sperm progressive motility, and sperm progressive+non-progressive motility were significantly lower in Group A and Group B than in Group C (P=0.000 in all cases); however, there were no differences between Group A and Group B regarding these parameters (Table 5). The sperm morphology index was significantly lower in Group A than in Group C (P=0.0024); no differences were found between Group B and Group C and Group B and Group A. The mean value of the sperm deformity index was significantly lower in Group A than in Group C P=0.004). Conclusion: Our study highlights the significant association between sperm morphology and male infertility in varicocele and idiopathic subfertile males.

Glucocorticoids improve sperm performance in physiological and pathological conditions: their role in sperm fight/flight response.

Glucocorticoids play a physiologic role in the adult male reproductive functions, modulating gonadal steroid synthesis and spermatogenesis, through the glucocorticoid receptor (GR). The expression of GR has been described in several key testicular cell types, including somatic cells and early germ cell populations. Nothing is known on GR in human spermatozoa. Herein, we explored the GR expression and its possible role in normal and testicular varicocele semen samples from volunteer donors. After semen parameter evaluation by macro- and microscopic analysis, samples were centrifuged; then spermatozoa and culture media were recovered for further investigations. By western blotting and immunofluorescence analyses we evidenced for the first time in spermatozoa the presence of GR-D3 isoform which was reduced in sperm from varicocele patients. By treating sperm with the synthetic glucocorticoid dexamethasone (DEXA), we found that survival, motility, capacitation, and acrosome reaction were increased in both healthy and varicocele samples. GR involvement in mediating DEXA effects, was confirmed by using the GR inhibitor mifepristone (M2F). Worthy, we also discovered that sperm secretes different cortisol amounts depending on its physio-pathological status, suggesting a defence mechanism to escape the immune system attach in the female genital tract thus maintaining the immune-privilege as in the testis. Collectively, our data suggests a role for glucocorticoids in determining semen quality and function, as well as in participating on sperm immune defensive mechanisms. The novelty of this study may be beneficial and needs to take into account in artificial insemination/drug discovery aimed to enhancing sperm quality.

Physico-morphological Characteristics of Semen Ejaculates in Vrindavani Crossbred Bulls

In crossbred cattle bulls, poor seminal quality and freezability are major constraints. The present study was conducted to determine semen quality parameters of Vrindavani crossbred bulls. In this study 130 semen samples were collected from five Vrindavani crossbred bulls. The volume, mass motility, concentration and individual progressive motility, viability and abnormality of spermatozoa were examined. Then ejaculates were categorized into four groups on the basis of spermatozoa concentration and individual progressive motility. The percentage of normozoospermia, asthenozoopsermia, oligozoospermia and oligoasthenozoospermia ejaculates were 44.62%, 29.23%, 11.54% and 14.61%, respectively. The overall mean of volume, mass motility, concentration of spermatozoa, individual progressive motility, viability and sperm abnormality in semen ejaculates of crossbred Vrindavani bulls were 4.64±0.63 mL, 2.48±0.09, 772.20±27.95 million/mL, 65.08±1.47%, 79.94±1.07% and 7.82±0.44%, respectively. Ejaculate mass motility, concentration, progressive motility and abnormality of spermatozoa varied significantly (p<0.05) among the semen ejaculate categories. The mean of the mass motility was significantly higher in normozoospermia as compared to other ejaculates categories. The normozoospermia and asthenozoospermia groups had significantly higher spermatozoa concentration than oligozoospermia and oligoasthenozoospermia groups. The normozoospermia ejaculates had significantly higher progressive motile spermatozoa as compared to other three ejaculates categories. Percent livability of spermatozoa in normozoospermia and oligozoospermia groups were significantly higher than asthenozoospermia and oligoasthenozoospermia groups. Percent abnormal spermatozoa on the other hand, was significantly lower in normozoospermia and oilgozoospermia groups than asthenozoopsermia and aligoasthenozoospermia groups.

Effect of the temperature of activation medium on fish sperm quality: Impact on fertilization in vitro in aquaculture practice

AbstractMotility is a key in spermatozoon function, determining semen quality and fertilizing capacity. Motility in fish spermatozoa occurs when they are diluted in a swimming solution, the adequacy of which is determined by factors which vary by species. Spermatozoon motility rate and velocity, as well as the duration of the motility period, are influenced by the temperature of the swimming medium. Increasing the temperature of the swimming medium has a significant negative effect on spermatozoa motility kinematics. It increases the metabolism rate of cells, causing a mismatch of energy resources and promoting changes in sperm movement. It also generates an increase in the production of reactive oxygen species (ROS) and thus oxidative stress, and moreover results in an inadequate capacity of antioxidant enzymes to protect the cell against the effects of oxidative stress induced by higher temperatures. The aim of this review is to update present knowledge about: mitochondria main source of ATP; the protein phosphorylation related to motility; epigenetic regulation of the temperature of the activation and the role of ion channels in regulation motility. These mechanisms involved in sperm motility are of vital importance in regulating fertilization, all of which can be influenced by the environment. We emphasize the importance of investigating spermatozoon mechanisms in depth, especially in the context of climate change which results in changing water temperatures. Our findings provide a better understanding of fish sperm physiology, and a biological foundation for the further development of spermatozoon motility investigation, and reproduction technologies.

The impact of magnetized drinking water on semen quality, fertility and hatchability rates of Fayoumi chicken

The present study was conducted to interpret the effect of magnetic field on water pH and calcium form, and evaluate semen quality, fertility and hatchability rates of Fayoumi chickens receiving magnetized drinking water (MDW). A total of 180 Fayoumi chickens (20 roosters + 160 hens), 54 weeks of age, were divided into two groups. The first group served as control, which received regular drinking water, and the second group was treated with MDW. Semen was collected twice (at day 15 and day 30 of the experiment) to determine semen quality. Fertility and hatchability were determined in two patches of hatching eggs through 1–15 and 16–30 days of the experiment. Results showed that the roosters of MDW group had higher live sperm and sperm motility ratios than those of the control group throughout the experimental period (1–30 days). The abnormal sperm ratio and methylene blue reaction time of MDW-received roosters were significantly lower than those of control roosters. Fertility and hatchability percentages of MDW group were greater compared to those of regular drinking water. Conclusively, alterations in the physiochemical characteristics of MDW and the calcium form can play a role in improving semen quality, fertility, and hatchability percentages during the late laying phase.

Oxidative Stress and Idiopathic Male Infertility.

Idiopathic male infertility (IMI) refers to the condition where semen quality declines, but exact causatives are not identified. This occurs in almost 30-40% of infertile men. Traditional semen analyses are extensively used for determining semen quality, but these bear critical shortcomings such as poor reproducibility, subjectivity, and reduced prediction of fertility. Oxidative stress (OS) has been identified as the core common mechanism by which various endogenous and exogenous factors may induce IMI. Male oxidative stress infertility (MOSI) is a term used to describe infertile males with abnormal semen parameters and OS. For the treatment of MOSI, antioxidants are mostly used which counteract OS and improve sperm parameters with appropriate combinations, dosage, and duration. Diagnosis and management of male infertility have witnessed a substantial improvement with the advent in the omics technologies that address at genetic, molecular, and cellular levels. Incorporation of oxidation-reduction potential (ORP) can be a useful clinical biomarker for MOSI. Moreover, various modulations of male fertility status can be achieved via stem cell and next-generation sequencing (NGS) technologies. However, several challenges must be overcome before the advanced techniques can be utilized to address IMI, including ethical and religious considerations, as well as the possibility of genetic abnormalities. Considering the importance of robust understanding of IMI, its diagnosis, and possible advents in management, the present article reviews and updates the available information in this realm, emphasizes various facets of IMI, role of OS in its pathophysiology, and discusses the novel concept of MOSI with a focus on its diagnostic and therapeutic aspects.

A Catalog of Human Genes Associated With Pathozoospermia and Functional Characteristics of These Genes.

Genetic causes of the global decline in male fertility are among the hot spots of scientific research in reproductive genetics. The most common way to evaluate male fertility in clinical trials is to determine semen quality. Lower semen quality is very often accompanied by subfertility or infertility, occurs in many diseases and can be caused by many factors, including genetic ones. The following forms of lowered semen quality (pathozoospermia) are known: azoospermia, oligozoospermia, asthenozoospermia, teratozoospermia, and some combined forms. To systematize information about the genetic basis of impaired spermatogenesis, we created a catalog of human genes associated with lowered semen quality (HGAPat) and analyzed their functional characteristics. The catalog comprises data on 126 human genes. Each entry of the catalog describes an association between an allelic variant of the gene and a particular form of lowered semen quality, extracted from the experimental study. Most genes included into the catalog are located on autosomes and are associated with such pathologies as non-obstructive azoospermia, oligozoospermia or asthenozoospermia. Slightly less than half of the included genes (43%) are expressed in the testes in a tissue-specific manner. Functional annotation of genes from the catalog showed that spermatogenic failure can be associated with mutations in genes that control biological processes essential for spermiogenesis (regulating DNA metabolism, cell division, formation of cellular structures, which provide cell movement) as well as with mutations in genes that control cellular responses to unfavorable conditions (stress factors, including oxidative stress and exposure to toxins).

Characteristics and Cryopreservation of Semen of Sex-Reversed Females of Salmonid Fish.

Sex reversal has been used as a breeding strategy by salmonid fish to produce genetically and phenotypically single sex populations. Production of all-female fish has great importance for the creation of monosex female triploids of salmonid fish, which are valued for their sterility, lack of female maturation, and larger commercial size. Among salmonids, the majority of rainbow trout (Oncorhynchus mykiss) production is based on all-female production with a high proportion of all-female triploid production in Europe. The main aim of this review is to present the recent knowledge regarding sex-reversed females (SRFs) of salmonid fish. We discuss the methods of sex reversal as well as their effects on the morphology and histology of the reproductive tract. We focus on the characteristics of SRF semen as well as the factors determining semen quality. The lower quality of SRF sperm compared to that of normal males has resulted in the need for the artificial maturation of semen. Most importantly, methods of semen storage—both short-term and long-term (cryopreservation)—that can improve hatchery operations are presented with the special emphasis on recent progress in development of efficient cryopreservation procedures and use of cryopreserved semen in hatchery practice. Moreover, we also address the emerging knowledge concerning the proteomic investigations of salmonid sperm, focusing primarily on the proteomic comparison of normal male and SRF testicular semen and presenting changes in SRF rainbow trout sperm proteome after in vitro incubation in artificial seminal plasma.

Localization of MIF-II on mammalian spermatozoa: A study revealing its structure, function and motility inhibitory pathway

Motility of spermatozoa is a crucial factor for determining semen quality. Here we report motility inhibitory factor (MIF-II) from goat epididymal plasma, revealing its structure, function, localization and motility inhibitory pathway. Structural characterization with MALDI revealed novelty of this protein while circular dichroism data confirmed its alpha helical nature. Higher dilutions of MIF-II antibody increased cauda sperm motility and induced immature/immotile caput sperm motility as tested microscopically. Higher number of sperm cells and lower dilutions of antibody induced agglutination in cauda sperm showing surface localization. Indirect immuno-fluorescence showed MIF-II localization throughout the caput sperm surface which relocated more towards acrosomal region with maturation. ELISA assay revealed gradual increase and decrease in concentration of MIF-II in epididymal plasma and plasma membrane respectively from caput to cauda. Signaling cascade that leads to sperm motility inhibition elevates nitric oxide levels through cAMP dependent pathway. MIF-II treatment doesn't alter sperm surface morphology. Expression pattern of MIF-II during epididymal maturation goes hand-in-hand with gaining motility potential as well as dormancy of spermatozoa before ejaculation. Both MIF-II and its antibody inhibit fertilization in-vitro thus expected to open new gateway for future male infertility and contraceptive development research.

Identification of genome regions determining semen quality in Holstein-Friesian bulls using information theory

Use of information theory can be an alternative statistical approach to detect genome regions and candidate genes that are associated with livestock traits. The aim of this study was to verify the validity of the SNPs effects on some semen quality variables of bulls using entropy analysis. Records from 288 Holstein-Friesian bulls from one AI station were included. The following semen quality variables were analyzed: CASA kinematic variables of sperm (total motility, average path velocity, straight line velocity, curvilinear velocity, amplitude of lateral head displacement, beat cross frequency, straightness, linearity), sperm membrane integrity (plazmolema, mitochondrial function), sperm ATP content. Molecular data included 48,192 SNPs. After filtering (call rate = 0.95 and MAF = 0.05), 34,794 SNPs were included in the entropy analysis. The entropy and conditional entropy were estimated for each SNP. Conditional entropy quantifies the remaining uncertainty about values of the variable with the knowledge of SNP. The most informative SNPs for each variable were determined. The computations were performed using the R statistical package. A majority of the loci had relatively small contributions. The most informative SNPs for all variables were mainly located on chromosomes: 3, 4, 5 and 16. The results from the study indicate that important genome regions and candidate genes that determine semen quality variables in bulls are located on a number of chromosomes. Some detected clusters of SNPs were located in RNA (U6 and 5S_rRNA) for all the variables for which analysis occurred. Associations between PARK2 as well GALNT13 genes and some semen characteristics were also detected.

Semen quality and infertility status can be identified through measures of oxidation-reduction potential.

Standard analyses for evaluating semen quality require technical expertise and are interpretive in nature. Oxidative stress (OS) alters many of the semen parameters; thus, a measure of OS could be an indicator of semen quality. Static oxidation-reduction potential (sORP) is a universal measure of OS traditionally used in environmental applications but is increasingly used in biomedical studies. sORP was measured to determine how well it associates with semen quality and if it differentiates semen from infertile patients and fertile donors. All study participants (Infertile, n=365 and Fertile, n=50) underwent standard semen analyses, and sORP was measured in unprocessed semen. In infertile patients, sORP increased with decreased total sperm number, motility and morphology. sORP values were higher in samples with abnormal quality (low number, motility and/or normal morphology) compared with those of normal quality. Infertile patients had higher sOPR values compared to fertile donors. A sORP cut-off value of 1.38mV/106 sperm/ml can differentiate normal from abnormal semen samples, while a cut-off value of 1.41mV/106 sperm/ml, can differentiate between infertile and fertile semen samples. In conclusion, sORP provides a quick and unbiased indicator of semen quality that can be a beneficial addition to semen analysis to determine semen quality and fertility status.

精子染色质扩散实验及吖啶橙染色检测精子DNA完整性研究

目的：采用精子染色质扩散(sperm chromatin dispersion, SCD)实验检测精子DNA碎片，分析比较SCD实验及吖啶橙染色实验(AOT)检测精子DNA完整性的应用价值。方法：对32例正常成年已生育的男性和27例特发性少精子症(idiopathic oligozoospermia, IO)患者同时进行SCD实验和AOT，对检测结果进行分析。在SCD实验中，DNA完整性正常无损伤精子的DNA扩散产生大晕环或中晕环，而DNA损伤产生DNA碎片的精子不产生或产生很小的晕环。AOT染色实验中，正常精子DNA为双链，染成绿色，不成熟或损伤精子DNA为单链，染成红色、橙色或黄色。结果：SCD实验检测结果为：IO患者精子大晕环、中晕环、小晕环和无晕环精子百分率分别平均为(51.8 ± 17.1)%、(9.4 ± 8.6)%、(11.9 ± 5.8)%和(26.7 ± 10.6)%，健康对照组平均为(73.2 ± 6.2)%、(14.8 ± 5.7)%、(6.8 ± 2.9)%及(5.3 ± 2.2)%，二组比较，差异有统计学意义(t = 4.256, P 0.05)。结论：精子DNA完整性异常可导致男性不育。SCD实验是一种有效的精子DNA完整性检测方法。 Objective: To analyze and compare the accuracy and utility of determining sperm DNA integrity by sperm chromatin dispersion (SCD) test and acridine orange staining test (AOT). Methods: The level of DNA fragmentation was determined by SCD test and AOT in 32 adult healthy fertile men (control group) and 27 idiopathic oligozoospermia (IO) patients. Sperm nuclei with large DNA dispersion halos or with medium-sized halos were normal and nuclei with small-sized halos or no halo were abnormal. Sperm DNA strand breaks were tested by AOT of green for native DNA and red for denatured DNA. Results: The mean percentage of sperm nuclei with large halos, medium-sized halos, small-sized halos and no halo in patients with IO was (51.8 ± 17.1)%, (9.4 ± 8.6)%, (11.9 ± 5.8)% and (26.7 ± 10.6)%, but in control group it was (73.2 ± 6.2)%, (14.8 ± 5.7)%, (6.8 ± 2.9)% and (5.3 ± 2.2)%, respectively, there was a significantly difference in the IO and control group (t = 4.256, P 0.05). Conclusion: The SCD is an effective test in sperm DNA fragmentation as a screening procedure to determine semen quality during basic infertility investigation for clinical use. The presence of sperm DNA fragmentation may lead to male infertility.

The Efficacy of In Vitro Sperm Tests in Predicting Pregnancy Success after Artificial Insemination in the Bitch

In many species, including the dog (Canis familiaris), semen analysis protocols has improved in recent years. In vitro assessment of semen quality provides important information; however, the ultimate parameter for determining semen quality is pregnancy outcome. The objective of the present study was to investigate the efficiency of in vitro sperm tests in predicting seminal quality in dogs by correlating those results with pregnancy success after artificial insemination (AI). Forty-eight semen samples refrigerated for 24, 48 or 72 hours were examined for sperm motility and progressive motility, membrane integrity, and zona pellucida-binding capacity before being used to artificially inseminate bitches. Results indicated positive correlations between pregnancy success and sperm motility (0.77), vigor (0.72), membrane integrity (0.72) and zona pellucida-binding (0.81). Both pregnancy success and the sperm characteristics declined with increased semen storage time. In conclusion, the semen characteristics evaluated in this study can be used to predict the conception potential of semen samples in the dog.

Effects of monochromatic light sources on sex hormone levels in serum and on semen quality of ganders

Light is an essential external factor influencing various physiological processes, including reproductive performance, in birds. Although several attempts have been made to understand the effect of light on poultry production, the effect of light of a particular wavelength (color) on the reproductive function in geese remains unclear. This study evaluated the effect of various monochromatic light sources on the levels of sex hormone and on semen quality of ganders. Of 30 male White Roman geese in their third reproductive season (average age=3years), 27 were divided into three groups receiving monochromatic white or red or blue lights. The birds were kept in an environmentally controlled house with a lighting photoperiod of 7L:17D for six weeks as the adaptation period. The photoperiod was subsequently changed to 9L:15D and maintained for 24 weeks. Three ganders at the beginning of the study and three from each group at the end of the adjusting period and the 20th and 30th week of the study period were sacrificed, and their testes and blood samples were collected for determining the sex hormone levels. Semen samples were collected for determining semen quality parameters, including the semen collection index, sperm concentration, semen volume, sperm motility, sperm viability, sperm morphology, and semen quality factor. The results showed that the testosterone and estradiol levels remained unchanged in all three groups at all time points. The ratio of testosterone to estradiol of ganders exposed to white light was significantly higher than that of ganders exposed to red light at the 30th week (P<0.05). Semen collection index and sperm viability of ganders exposed to blue light were significantly the lowest (P<0.05). Moreover, sperm motility, sperm viability, and percentage of morphologically normal spermatozoa of ganders in white light were the highest (P<0.05). In conclusion, the results of this study suggested that artificial illumination with white light may maintain a better semen quality than that with red or blue lights in ganders.

Kualitas semen dan produksi semen beku pada bangsa sapi dan bulan penampungan yang berbeda

The purpose of this research was to determine semen quality and frozen semen production of beef cattle at different month collection in Lembang Artificial Insemination Center. Beef cattle strain that used for this research are Aceh, Angus, Brahman, Limousine, Madura, Ongole and Simmental. The research parameter included semen volume, pH, motility of spermatozoa, concentration of spermatozoa, number of spermatozoa, number of spermatozoa motil and number of frozen semen straw. The method of this research using factorial completely Randomized Design and each had 3 replication per treatment. The different of beef cattle have given a significant effect of semen volume (P 0.05). Different month collection have a significant influence of semen volume (P<0.01), pH (P<0.05), motility of spermatozoa (P<0.01), concentration of spermatozoa (P<0.01), number of spermatozoa (P<0.01), number of spermatozoa motil (P<0.01). Interaction between breed of beef cattle and different month of collection have given a significant influence of semen volume (P<0.01), motility spermatozoa (P<0.01), concentration spermatozoa (P<0.05) and number of spermatozoa motil (P<0.01). Keywords : sperm motility, concentration, number of motile spermatozoa

Expression of matrix metalloproteinases (MMP-2, MMP-9) and their inhibitors (TIMP-1, TIMP-2) in canine testis, epididymis and semen.

This study aims to investigate the role of matrix metalloproteinases (MMPs) in determining semen quality and to evaluate the expression and cellular localization of MMP-2, MMP-9, tissue inhibitor of metalloproteinase-1 (TIMP-1) and TIMP-2 in the testes, epididymis and ejaculated spermatozoa. Gelatinase activities between normal (n=21) and abnormal (n=25) semen samples showed a significant, sixfold increase in proMMP-2 and MMP-2 activity in high than low sperm concentration samples (p<0.001). ProMMP-9 and MMP-9 levels were significantly elevated in samples with low sperm counts compared to those with high sperm density (p<0.001). High levels of proMMP-2 and MMP-2 were associated with high sperm motility (≥70%, p<0.001). Sperm-rich fraction showed significantly (eight-fold) higher proMMP-9 enzymatic activity compared with prostatic fraction. The mRNA expressions of MMP-2, MMP-9, TIMP-1 and TIMP-2 were confirmed in testicular and epididymal tissues. Immunohistochemical staining illustrated the MMP-2-specific strong immunoreactivity in the head of mature spermatids during spermatogenesis, whereas MMP-9, TIMP-1 and TIMP-2 were absent in these cells. Matrix metalloproteinase-9 immunoreactivity was observed in the spermatocyte and round spermatid, whereas TIMP-1 was only exhibited in the residual bodies. Immunolabeling of epididymal and ejaculated sperm demonstrated MMP-2 localization along acrosomal region of sperm, while MMP-9, TIMP-1 and TIMP-2 localization was merely limited to the flagella. In conclusion, spermatozoa initially acquire MMP-2 during their formation at testicular level, and the presence of this protein persists through the epididymal transit and up to ejaculate. The enzymatic activity of MMP-2 and MMP-9 may serve as an alternative biomarker in determining semen quality.

Pig Spermatozoa Defect in Acrosome Formation Caused Poor Motion Parameters and Fertilization Failure through Artificial Insemination and In vitro Fertilization.

The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not sufficient to select normal and fertilizable spermatozoa. In conclusion, the present study suggests that the acrosome staining and detailed motion characteristics such as progressiveness, VCL, and VSL should be included in determining semen quality together with primary parameters for successful AI and high breeding performance in the swine industry.

A re-evaluation of sperm ultrastructure in the emu, Dromaius novaehollandiae

Existing reports on sperm structure in the emu do not adequately illustrate or describe all the salient ultrastructural features necessary for a meaningful comparison of normal and abnormal sperm in this species. As sperm morphology forms an important parameter in determining semen quality, and in view of the proposed role of artificial insemination in the farming of ratites, this article re-evaluates and complements the existing data on the topic, provides a fully illustrated description of emu sperm ultrastructure, and documents some unreported morphologic features. Conventional transmission and scanning electron microscopy and high resolution scanning electron microscopy were used to describe the ultrastructure of sperm harvested from the distal deferent duct of sexually mature birds slaughtered during the breeding season. In addition to broadly confirming the basic ultrastructural characteristics previously described for emu sperm, this study revealed a number of unreported morphologic features. These included distinct differences in surface properties between the acrosome and nucleus, the presence of a thread-like appendage near the base of the nucleus, variable positioning of the annulus relative to structures located at the midpiece–principal piece junction and regional differentiation of the principal piece. Although the emu displayed similar basic morphologic features to sperm of other ratites and the tinamou, marked structural peculiarities were obvious, notably the lack of an endonuclear canal and a perforatorium and the presence of significantly more mitochondria in the midpiece coupled with an absence of intermitochondrial cement. Although the broad morphologic features of emu sperm would appear to add credence to the general view that the ratites, together with the tinamous, form a monophyletic group at the base of the avian phylogenetic tree, it is also clear that emu sperm are distinctly different from those of the ostrich, rhea, and tinamou which together share morphologic affinities. This observation may lend some support to the alternate view that the Australasian ratites represent a separate clade that developed independently from flightless ancestors.

The cryoprotective effects of amino acids supplementation on cooled and post-thaw Markhoz bucks semen quality

Oxidative damage to sperm resulting from reactive oxygen species generated by the cellular components of semen is one of the main causes for the decline in motility and fertility of sperm during the freeze–thawing process. This study has been designed to evaluate the effects of glutamine, proline, Basal Medium Eagle (BME) and creatine on cooled/rewarmed and frozen-thawed Markhoz bucks spermatozoa. A total number of 42 ejaculates were collected twice a week from 4 mature bucks using an artificial vagina. Only semen samples with motility of more than 70% and sperm concentrations higher than 3×109sperm/ml were used for cryopreservation. After initial evaluation, the collected ejaculates were pooled together to minimize individual variation. Twelve pooled ejaculates were included in the study. The pooled semen samples were diluted 1:4 before cooling/rewarming (part 1) and freezing/thawing (part 2) with Tris-Citrate-Froctose-Yolk extender containing creatine (2.5, 5 or 7.5mM), glutamine (5mM), proline (5mM) and BME (10%). Control extender was not supplemented with amino acids and creatine. To determine semen quality characteristics, the rates of motility, progressive motility, viability, total abnormality and acrosome and membrane integrity were evaluated. The obtained results indicated that the addition of creatine (2.5mM), glutamine (5mM), proline (5mM) or BME (10%) significantly increase (p<0.05) sperm parameters (motility, progressive motility, viability, acrosome and membrane integrity) and decrease total abnormalities compared to control extender. However, extenders containing creatine (2.5mM) or creatine (2.5mM) plus BME (10%) presented significantly higher values (p<0.05) than other treatments. In conclusion, the results presented in this study show that using antioxidants can improve the sperm characteristics of spermatozoa before and after freezing.

Endocannabinoids as Markers of Sperm Quality: Hot Spots

Endocannabinoids as Markers of Sperm Quality: Hot Spots