Affect Sperm Motility Research Articles

Chronic exposure to organic oxygen-demanding pollutants at an environmentally realistic concentration affects sperm motility in zebrafish

Wastewater and organic oxygen-demanding pollutants (ODPs) are produced by various factories in China, the United States and other countries. However, whether ODP affects reproductive health remains unclear. To investigate the impact of environmental concentrations of ODP exposure on reproductive health, adult male zebrafish were used to evaluated the effects ODP exposure on the fertility in this study. We found that exposure to ODP reduced the sperm motility of adult male zebrafish. Similarly, the testosterone content of the experimental zebrafish was obviously decreased. Transcription of immune response-related genes, including tumor necrosis factor (tnf)-α, il-1β, and il-8, was upregulated upon exposure to ODP. Mating experiments indicated that the hatching time of the offspring embryos was clearly prolonged upon ODP exposure, but the embryo fertilization rate was not different. These results assumed that exposure to ODP at ambient concentrations visibly affected the sperm motility in adult zebrafish maybe due to the expression of immune response-related genes in the zebrafish male gonads and the release of pro-inflammatory mediators. Therefore, we assumed that the impact of ODP on the reproductive health of aquatic organisms cannot be ignored.

Bovine sperm-oviduct interactions are characterized by specific sperm behaviour, ultrastructure and tubal reactions which are impacted by sex sorting

To date sperm-oviduct interactions have largely been investigated under in vitro conditions. Therefore we set out to characterize the behaviour of bovine spermatozoa within the sperm reservoir under near in vivo conditions and in real-time using a novel live cell imaging technology and a newly established fluorescent sperm binding assay. Sperm structure and tubal reactions after sperm binding were analysed using scanning and transmission electron microscopy and histochemistry. As a model to specify the impact of stress on sperm-oviduct interactions, frozen-thawed conventional and sex-sorted spermatozoa from the same bulls (n = 7) were co-incubated with oviducts obtained from cows immediately after slaughter. Our studies revealed that within the oviductal sperm reservoir agile (bound at a tangential angle of about 30°, actively beating undulating tail), lagging (bound at a lower angle, reduced tail movement), immotile (absence of tail movement) and hyperactivated (whip-like movement of tail) spermatozoa occur, the prevalence of which changes in a time-dependent pattern. After formation of the sperm reservoir, tubal ciliary beat frequency is significantly increased (p = 0.022) and the epithelial cells show increased activity of endoplasmic reticula. After sex sorting, spermatozoa occasionally display abnormal movement patterns characterized by a 360° rotating head and tail. Sperm binding in the oviduct is significantly reduced (p = 0.008) following sexing. Sex-sorted spermatozoa reveal deformations in the head, sharp bends in the tail and a significantly increased prevalence of damaged mitochondria (p < 0.001). Our results imply that the oviductal cells specifically react to the binding of spermatozoa, maintaining sperm survival within the tubal reservoir. The sex-sorting process, which is associated with mechanical, chemical and time stress, impacts sperm binding to the oviduct and mitochondrial integrity affecting sperm motility and function.

Sphingomyelin Synthase 2 Participate in the Regulation of Sperm Motility and Apoptosis.

Sphingomylin participates in sperm function in animals, and also regulates the Akt and ERK signaling pathways, both of which are associated with the asthenospermia. Sphingomyelin synthase 2 (SMS2) is involved in the biosynthesis of sphingomylin. To determine the relationship between SMS2 and human sperm function, we analyzed the distribution of SMS2 in human sperm and testes, and SMS2 expression in patients with asthenospermia and normozoospermia; human sperm were treated with anti-SMS2, and the sperm motility, penetration ability into methylcellulose, capacitation and acrosome reaction, and sperm [Ca2+]i imaging were evaluated, while the Akt and ERK pathway and cleaved caspase 3 were also analyzed. Results showed that SMS2 was localized in the testis and human sperm, and the protein levels of normozoospermia were higher than asthenospermia. Inhibition of SMS2 activity significantly decreased sperm motility and penetration ability into methylcellulose, but had no influence on capacitation and acrosome reaction, or on intracellular [Ca2+]i compared to IgG-treated control groups. Moreover, the phosphorylation level of Akt was decreased, whereas the phosphorylation of ERK and cleaved-caspase 3 levels were significantly increased. Taken together, SMS2 can affect sperm motility and penetration ability into methylcellulose, and participate in apoptosis associated with the Akt and ERK signaling pathways.

Red-Light Irradiation of Horse Spermatozoa Increases Mitochondrial Activity and Motility through Changes in the Motile Sperm Subpopulation Structure.

Previous studies in other mammalian species have shown that stimulation of semen with red-light increases sperm motility, mitochondrial activity, and fertilizing capacity. This study sought to determine whether red-light stimulation using a light emitting diode (LED) at 620–630 nm affects sperm motility and structure of motile subpopulations, sperm viability, mitochondrial activity, intracellular ATP levels, rate of O2 consumption and DNA integrity of horse spermatozoa. For this purpose, nine ejaculates were collected from nine different adult stallions. Upon collection, semen was diluted in Kenney extender, analyzed, its concentration was adjusted, and finally it was stimulated with red-light. In all cases, semen was packaged in 0.5-mL transparent straws, which were randomly divided into controls and 19 light-stimulation treatments; 6 consisted of a single exposure to red-light, and the other 13 involved irradiation with intervals of irradiation and darkness (light-dark-light). After irradiation, sperm motility was assessed using a Computerized Semen Analysis System (CASA). Flow cytometry was used to evaluate sperm viability, mitochondrial membrane potential and DNA fragmentation. Intracellular levels of ATP and O2 consumption rate were also determined. Specific red-light patterns were found to modify kinetics parameters (patterns: 4, 2-2-2, 3-3-3, 4-4-4, 5-1-5, and 5-5-5 min), the structure of motile sperm subpopulations (patterns: 2, 2-2-2, 3-3-3, and 4-1-4 min), mitochondrial membrane potential (patterns: 4, 3-3-3, 4-4-4, 5-1-5, 5-5-5, 15-5-15, and 15-15-15 min), intracellular ATP levels and the rate of O2 consumption (pattern: 4 min), without affecting sperm viability or DNA integrity. Since the increase in some kinematic parameters was concomitant with that of mitochondrial activity, intracellular ATP levels and O2 consumption rate, we suggest that the positive effect of light-irradiation on sperm motility is related to its impact upon mitochondrial activity. In conclusion, this study shows that red LED light stimulates motility and mitochondrial activity of horse sperm. Additional research is needed to address the impact of red-light irradiation on fertilizing ability and the mechanisms through which light exerts its effects.

Ion-rich potash mining effluents affect sperm motility parameters of European perch, Perca fluviatilis, and impair early development of the common roach, Rutilus rutilus

Secondary salinization of freshwater ecosystems is of increasing global concern. One of the main causes are the effluents of the potash mining industry containing high concentrations of major ions (Cl−, Na+, Mg2+, K+). In Germany, the ongoing discharge of effluents into the River Werra led to a strong impoverishment of the biodiversity and abundance of local species. Young cohorts of many freshwater fish are completely absent suggesting reproductive failure under these conditions. Therefore, the aim of the study was to experimentally investigate the effects of high concentrations and imbalances of ions that are prevalent in potash mining effluents on reproductive traits of native freshwater teleosts. Sperm motility parameters of the common roach, Rutilus rutilus, and European perch, Perca fluviatilis, were assessed as well as fertilization rate, egg size, hatching, malformations and mortality of embryonic and larval stages of roach. Concentrations of the permitted thresholds (HT) and future thresholds (LT) as well as three ion solutions containing high Mg2+ (Mg), high K+ (K) and both in combination (Mg + K) were tested. Curvilinear velocity and linearity of perch spermatozoa were elevated with potentially adverse effects on fertilization success. Sperm motility parameters and fertilization rate of roach were not affected. However, egg sizes of roach were increased in all groups due to the osmotic action of ions and in LT, premature hatch was observed. Furthermore, all groups comprised a higher number of malformations including pericardial edema and spine curvatures and group HT exhibited a higher mortality rate compared to control. The results clearly demonstrated that particularly the sum of high concentrations of ions, as prevalent in HT and LT, rather than individual ion species exerts detrimental effects on early development of roach potentially increasing overall mortality under natural conditions. These results emphasize that currently permitted and future thresholds are exceeding tolerated ion concentrations.

Validation of the Turkey Semen Cryopreservation by Evaluating the Effect of Two Diluents and the Inseminating Doses.

Simple SummaryAchieving an effective freezing protocol, that is able to preserve the fertilizing ability of turkey semen, is a key aim for the establishment of the first national semen cryobank of autochthonous chicken and turkey breeds within our national project (Tutela della biodiversità nelle razze avicole italiane—TuBAvI). In this regard, we have performed different studies in order to define the best conditions for cryopreservation of turkey semen; namely, we identified an effective freezing protocol which is based on the use of dimethylsulfoxide as a permeant cryoprotectant (CPA) combined with Ficoll as a non-permeant CPA. Here, our purpose was to test this protocol in vivo, by evaluating the effect of two extenders and three inseminating doses. The good fertility and hatching rates achieved here are promising for future studies, in which our cryopreservation protocol will be tested on Italian autochthonous turkey breeds and also to the advantages offered by the extensive use of frozen semen in the turkey breeding industry.This study was designed to test the fertilizing ability of cryopreserved turkey semen, and here, two experiments were performed: an in vitro analysis to assess the effects of Tselutin and Lake diluents and an in vivo test to determine the fertility and hatching rates by also studying the feat of three insemination doses (250, 400 and 600 × 106 sperm/hen). Pooled semen samples were diluted with Tselutin or Lake extender which contained 20% of dimethylsulfoxide and 1 mM of Ficoll at final sperm concentration of 3 × 109 sperm/mL. Thereafter, semen was packaged into straws and frozen on liquid nitrogen. The post-thaw sperm quality was evaluated considering motility (computer-aided sperm analysis—CASA system) and membrane integrity (flow cytometry). Significantly higher values of progressive motility and some kinetic parameters in semen frozen with Lake were found. When we compared the extenders in vivo, no significant effects were detected, whilst sperm concentration significantly affected both fertility and hatching rates, with the best results obtained with the sperm concentration of 400 × 106 sperm/hen. From the results obtained, it emerged that the extender type only affected sperm motility characteristics, not the fertilizing ability of frozen-thawed semen, while inseminating dose markedly affected fertility and hatching rates.

Role of Exosomes in the Exchange of Spermatozoa after Leaving the Seminiferous Tubule: A Review.

Exosomes are extracellular vesicles (EVs) released from cells upon fusion of an intermediate endocytic compartment with the plasma membrane. They refer to the intraluminal vesicles released from the fusion of multivesicular bodies with the plasma membrane. The contents and number of exosomes are related to diseases such as metabolic diseases, cancer and inflammatory diseases. Exosomes have been used in neurological research as a drug delivery tool and also as biomarkers for diseases. Recently, exosomes were observed in the seminal plasma of the one who is asthenozoospermia, which can affect sperm motility and capacitation. The main objective of this review is to deeply discuss the role of exosomes in spermatozoa after leaving the seminiferous tubule. We conducted an extensive search of the literature available on relationships between exosomes and exosomes in spermatozoa on the bibliographic database. This review thoroughly discussed the role that exosomes play in the exchange of spermatozoa after leaving the seminiferous tubule and its potential as a drug delivery tool and biomarkers for diseases as well.

Vanadium adversely affects sperm motility and capacitation status via protein kinase A activity and tyrosine phosphorylation

Vanadium is a chemical element that enters the atmosphere via anthropogenic pollution. Exposure to vanadium affects cancer development and can result in toxic effects. Multiple studies have focused on vanadium’s detrimental effect on male reproduction using conventional sperm analysis techniques. This study focused on vanadium’s effect on spermatozoa following capacitation at the molecular level, in order to provide a more detailed assessment of vanadium’s reproductive toxicity. We observed a decrease in germ cell density and a structural collapse of the testicular organ in seminiferous tubules during vanadium treatment. In addition, various sperm motion parameters were significantly decreased regardless of capacitation status, including sperm motility, rapid sperm motility, and progressive sperm motility. Curvilinear velocity, straight-line velocity, average path velocity, beat cross frequency, and mean amplitude of head lateral displacement were also decreased after capacitation. Capacitation status was altered after capacitation. Vanadium dramatically enhanced protein kinase A (PKA) activity and tyrosine phosphorylation. Taken together, our results suggest that vanadium is detrimental to male fertility by negatively influencing sperm motility, motion kinematics, and capacitation status via abnormal PKA activity and tyrosine phosphorylation before and after capacitation.

Oxidative stress in cryopreserved semen of sex-reversed female and normal male rainbow trout

The aim of the current study was to determine the sperm quality of sex-reversed female (SRF) and normal male rainbow trout measured as ROS+ (percentage of spermatozoa exhibiting reactive oxygen species) production and TAC (total antioxidant capacity) in semen cryopreserved at different glucose concentration [optimal (0.15 M), low (0.11 M) and high (0.19 and 0.21 M for SRF and normal male rainbow trout, respectively)] in 7.5% methanol (glucose-methanol extender). Our hypothesis assumes that at non-optimal glucose concentrations, production of ROS+ is induced, resulting in damage to spermatozoa which should be characteristic for normal males and SRF rainbow trout. Such approach would allow to describe the characteristics of the response of spermatozoa of normal males and SRF to oxidative stress which is prerequisite for better understanding the mechanism of oxidative stress of normal males and SRF. This knowledge can be important for improvement of protocols of short-term storage and cryopreservation of SRF and normal males semen. We established baseline values for ROS+ and TAC for fresh semen from SRF and normal male rainbow trout and found a negative regression between those two characteristics in SRF. We determined the effect of glucose concentration on ROS+ production, TAC and sperm quality parameters of cryopreserved semen. TAC activity was monitored at the same time, and its relationship to oxidative stress was assessed. The glucose concentration significantly affected sperm motility, curvilinear velocity and viability after thawing. Cryopreservation resulted in increased ROS+ values in both tested groups. On the other hand, TAC values were significantly lower in cryopreserved samples than in equilibrated samples of SRF rainbow trout semen at all tested glucose concentrations. Our results strongly suggest that increasing the osmolality of the extender induces oxidative stress in spermatozoa of SRF and normal male rainbow trout. Most changes are related to freezing and thawing; however, some changes may occur during equilibration. Specific changes in oxidative stress characteristics occur in fresh and cryopreserved semen of SRF and normal males. An inverse relationship between ROS+ and TAC is likely caused by consumption of antioxidants by ROS+. Our results can be useful for optimization of cryopreservation protocols for salmonid fish. Further studies are necessary to elucidate the mechanisms that lead to differences in oxidative stress levels in semen from SRF and normal male rainbow trout.

Effect of semen extenders on viability of ISA Brown and Hubbard Flex roosters' sperm stored for 24 h

Artificial insemination is used in almost 95% of turkey reproductive flocks and is becoming more important in chickens, particularly broiler breeders, as well as in assisted reproduction of wild birds kept in breeding centers. Diluted semen is recommended for artificial insemination. Pooled semen samples collected twice a week by dorso-abdominal massage from 2 chicken lines: laying—ISA Brown (ISA-B) and meat type—Hubbard Flex (H-F) were divided into 5 parts: neat semen and diluted in 1:2 ratio with 4 extenders: basic EK; EK + 1 μg/mL organic selenium and 8 μg/mL vitamin E; EK + 10 mg/mL of royal jelly; and EK + 0.25 g/mL of lyophilized bovine colostrum. Diluted semen samples were evaluated after 15 min and then 24 h storage at 4°C. Sperm concentration, motility, motility parameters (with Sperm Class Analyzer), and morphology were evaluated in the neat semen, whereas in diluted and stored samples, the last 3 traits were determined. In case of both lines, dilution did not affect (P > 0.05) the number of live normal cells (78.0–81.1% in ISA Brown and 73.8–68.7% in Hubbard Flex) in relation to neat semen; however, bovine colostrum addition increased (P < 0.05) the percentage of bulb head sperm (5.7 vs. 10.0% and 12.1 vs. 17.6%, for ISA and Hubbard, respectively) and decreased sperm motility (67.4 vs. 92.9% and 67.3 vs. 98.5% for ISA and Hubbard). The 24 h storage of neat semen and semen diluted with colostrum caused (P < 0.05) the unfavorable changes in all evaluated traits and both chicken lines, whereas semen dilution with remaining extenders decreased the percentage of live normal cells (by 18.8–23.4% ISA and by 20.9–25.5% Hubbard) but did not affect sperm motility (81.5–87.6% for ISA and 81.1–96.6% for Hubbard). Sperm motility and motility parameters depended both on the extender and chicken line.

Effect of caspase inhibitor Z-VAD-FMK on bovine sperm cryotolerance.

The aim of this study was to evaluate the treatment of bovine semen with the pan-caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (Z-VAD-FMK), before or after freezing on semen quality. After the initial assessment, sperm from 4 bulls were pooled (Experiment 1) and cryopreserved in BioXcell containing 0, 20 and 100μM Z-VAD-FMK. After thawing semen viability, motility, membrane integrity, as well as DNA fragmentation and ΔΨm were evaluated. In Experiment 2, bovine frozen/thawed sperm were incubated for 1hr with 0, 20 and 100µM Z-VAD-FMK before assessing the semen quality. The treatment with Z -VAD-FMK before cryopreservation improved post-thawing sperm motility compared to the control group (p<.05), while no differences were recorded in sperm viability and membrane integrity among groups (on average 86.8±1.5 and 69.1±1.4, respectively). Interestingly, at the highest concentration, DNA fragmentation decreased (p<.05), while the percentage of spermatozoa with high ΔΨm increased (p<.05). The results of Experiment 2 showed that 1-hr treatment with Z-VAD-FMK did not affect sperm motility and viability (on average 63.4±5.8 and 83.7.1±1.2, respectively). However, Z-VAD-FMK improved sperm membrane integrity (p<.05) and at the highest concentration tested decreased the proportion of sperm showing DNA fragmentation (p<.05). No differences were recorded in the percentage of spermatozoa with high ΔΨm (on average 57.0±11.4). In conclusion, the treatment with 100µM of the caspase inhibitor Z-VAD-FMK before freezing increased bovine sperm mass motility and ΔΨm, while decreasing sperm DNA fragmentation. Treatment of semen after thawing with 100µM Z-VAD-FMK improved sperm membrane integrity and reduced DNA fragmentation.

Effects of glucose, methanol concentration, and time of equilibration on post-thaw sperm motility of rainbow trout semen

The objective of this work was to develop a complex standardisation of cryopreservation procedure for rainbow trout semen. The specific objectives were to determine the effects of the final glucose concentration (0.11–0.23 M), final methanol concentration in the extender (0–20%), equilibration time (5–180 min), and post-thaw storage time of semen (0–180 min) on sperm motility parameters. The final glucose and methanol concentrations significantly affected sperm motility after thawing. The highest sperm motility (63–75%) was found for glucose concentrations ranging from 0.13 to 0.17 M and for 5.0 to 10.0% methanol, with the highest motility (70%) at 7.5% methanol. Concerning post-thaw storage equilibration times, sperm motility of the equilibrated semen was not affected by 120 min, while a significant decrease was recorded after 180 min of equilibration. The highest values of sperm motility immediately after thawing were observed in samples equilibrated for 5 to 120 min. The sperm motility after thawing was not significantly affected up to 60 min for samples equilibrated for 5, 15, and 30 min. However, for samples equilibrated for 60, 120, and 180 min, significant decreases were observed at 60 min after thawing. Our results indicate the importance of the standardisation of concentrations of cryoprotectants as well as cryopreservation equilibration time. The recommended cryopreservation procedure includes final sperm concentration (1.0 × 109 spermatozoa ml−1), as we determined previously, 0.15 M glucose, 7.5% methanol, 15 min equilibration time, and post-thaw storage up to 60 min. The developed standardised procedure has potential for the cryopreservation of rainbow trout semen for practical purposes.

MiR-26a is Involved in Glycometabolism and Affects Boar Sperm Viability by Targeting PDHX.

miR-26a is associated with sperm metabolism and can affect sperm motility and apoptosis. However, how miR-26a affects sperm motility remains largely unknown. Our previous study indicated that the PDHX gene is predicted to be a potential target of miR-26a, which is responsible for pyruvate oxidative decarboxylation which is considered as a key step for connecting glycolysis with oxidative phosphorylation. In this study, we first reported a potential relationship between miR-26a and PDHX and their expressions in fresh, frozen-thawed, and epididymal boar sperm. Then, sperm viability and survival were determined after transfection of miR-26a. mRNA and protein expression level of PDHX in the liquid-preserved boar sperm after transfection were also determined by RT-qPCR and Western Blot (WB). Our results showed that expression level of PDHX was significantly increased during sperm transit from epididymal caput to corpus and cauda. Similarly, expression of PDHX was significantly higher (P < 0.05) in fresh sperm as compared to epididymal cauda and frozen-thawed sperm. However, the expression of miR-26a in epididymal corpus sperm was significantly higher (P < 0.05) than that of caput and cauda sperm. Furthermore, after transfection of boar sperm with miR-26a mimic and inhibitor under liquid storage, the lowest and highest sperm viability was observed in miR-26a mimic and inhibitor treatment (P < 0.05), respectively. The protein levels of PDHX, after 24 and 48 h of transfection of miR-26a mimics and inhibitor, were notably decreased and increased (P < 0.05), respectively, as compared to negative control (NC) group. In conclusion, the novel and enticing findings of our study provide a reasonable evidence that miR-26a via PDHX, a link between glycolysis and oxidative phosphorylation, could regulate the glycometabolic pathway which eventually affect boar sperm viability and survival.

Single-Layer Colloid Centrifugation as a Method to Process Urine-Contaminated Stallion Semen After Freezing-Thawing

Urospermia is a major ejaculatory dysfunction affecting stallions. It has been thought that urine-contaminated semen should not be cryopreserved; however, on select cases, urine contamination of semen cannot be avoided. A recent study suggested that urospermic semen can be cryopreserved after cushion centrifugation and extension. Thus, this study aimed to assess the use of single-layer colloid centrifugation (SLC) to process frozen-thawed urine-contaminated stallion semen. Raw ejaculates (n = 55) from eight stallions were split into three groups: no urine, low (20%), or high (50%) urine contamination. Semen was extended 1:1, cushion-centrifuged, and resuspended at 200 million sperm/mL in BotuCrio. Resuspended semen was loaded in 0.5 mL straws and cryopreserved in liquid nitrogen. Samples were thawed (37°C for 30 seconds) and processed by SLC (400 g/30 minutes). Percentages of total motility (TM) and progressive motility (PM) were assessed with computer-assisted semen analyzer. Sperm viability (%VIAB) and yield were assessed with a NucleoCounter before and after gradient centrifugation. Data were analyzed with two-way ANOVA and Tukey’s test. The motility parameters TM before SLC (control: 35 ± 2; low: 33 ± 0.7; high: 22 ± 1.8) after SLC (control: 51 ± 3.6; low: 42 ± 2.2; high: 25 ± 2.8) and PM before SLC (control: 24 ± 1.8; low: 21 ± 1.14; high: 12 ± 1.5) and after SLC (control: 40.3 ± 3.2; low: 31 ± 3.9; high: 14 ± 2) significantly decreased with increasing urine contamination. Urine contamination marginally reduced (P < .05) sperm viability after cryopreservation before SLC (control: 45 ± 0.7; low: 27 ± 0.2; high: 27 ± 0.3) and after SLC (control: 54 ± 0.5; low: 49 ± 0.7; high: 38 ± 0.6). Recovery rates of sperm after centrifugation were not significantly different between groups. In conclusion, urine contamination affects sperm motility parameters in a dose-dependent manner. Post-thaw SLC selected sperm with higher motility and viability in control and low groups but only selected sperm with higher viability in the high group.

Hepatitis B virus surface protein induces oxidative stress by increasing peroxides and inhibiting antioxidant defences in human spermatozoa.

Hepatitis B virus (HBV) infection may affect sperm motility in patients with HBV. HBV surface protein (HBs) decreases mitochondrial membrane potential, impairs motility and induces apoptotic-like changes in human spermatozoa. However, little is known about how human spermatozoa respond to reactive oxygen species (ROS; mainly peroxides) induced by HBs. In this study, HBs induced supraphysiological ROS levels in human spermatozoa and reduced the formation of 2-cell embryos (obtained from hamster oocytes and human spermatozoa). HBs induced a pre-apoptotic status in human spermatozoa, as well as antioxidant defences by increasing glutathione peroxidase 4 (GPX4) and peroxiredoxin 5 (PRDX5) levels. These results highlight the molecular mechanism responsible for the oxidative stress in human spermatozoa exposed to HBV and the antioxidant defence response involving GPX4 and PRDX5.

A model protocol for the cryopreservation and recovery of motile lizard sperm using the phosphodiesterase inhibitor caffeine.

Reproductive technologies such as genome storage and assisted reproduction have a significant role to play in ending or reversing species extinctions. However, such technologies for non-model organisms (i.e. non-mammalian species) are poorly developed. This is particularly true for the reptiles, in which there is a dearth of successful protocols for cryopreserving reptile spermatozoa, despite limited attempts. We investigated sperm cryopreservation in the Australian lizard Varanus panoptes with the objective of addressing the unmet need for an optimized cryopreservation protocol for the spermatozoa of squamate reptiles. We tested the efficacy of two cryoprotectants [dimethyl sulfoxide (DMSO) and glycerol] as well supplementation with a phosphodiesterase inhibitor (caffeine) to promote post-thaw motility. For cryopreservation, sperm were cooled in straws suspended in liquid nitrogen vapour for 5 minutes (approximately −135°C), before being plunged into liquid nitrogen (approximately −196°C), and later thawed in a water bath at 35°C. Samples were incubated post-thaw for 10 minutes in the presence or absence of 10 mM of caffeine. Both cryoprotectant type and concentration significantly affected percent sperm motility pre-freezing, with DMSO being less cytotoxic than glycerol and motility decreasing at higher concentrations of both cryoprotectant types. While cold shock did not significantly affect sperm motility, both cryoprotectant type and concentration did significantly impact the motility of post-thawed spermatozoa. Thus, mid-range concentrations (10% v/v) of DMSO and glycerol yielded a greater post-thaw motility compared with 5 and 20% v/v, while DMSO proved superior to glycerol. The addition of caffeine resulted in a significant recovery of post-thaw motility for both cryoprotectants, with higher rates of motility being associated with higher cryoprotectant concentrations. These protocols provide a significant step forward for in situ and ex situ management of threatened reptiles and add to recent evidence that reptilian sperm may have the full range of phosphorylation-mediated cellular mechanisms associated with capacitation, motility and metabolic regulation found in mammalian sperm.

The Effects of Distance and Duration of Exposure to Electromagnetic Radiation of Hair Dryer on Rat Sperm Parameters

Background: As a part of modern life, using electromagnetic field exposure has increased. EMFs might produce a variety of adverse effects on different parts of human body including the reproductive system. The consequences of exposure to the emitted EMFs from a hair dryer on sperm parameters were the major concerns of the present study. Materials and Methods: Fifty four adult Male Sprague-Dawley rats were randomly assigned to 9 groups. Samples from all experimental groups received exposure with two different modes: switch on (hot wind) and switch off (cold wind). Samples of group 1 (control) maintained under experimental conditions without any exposure. The exposure protocol for samples in groups 2 to 9 were 10 cm at 10 min—on, 10 cm at 10 min—off, 10 cm at 20 min—on, 10 cm at 20 min—off, 20 cm at 10 min—on, 20 cm at 10 min—off, 20 cm at 20 min—on, 20 cm at 20 min—off. After exposure sperm motility was analyzed with a light microscope as a blind experiment. Nonparametric tests were used to detect statistically significant differences between different groups. Results: Findings of our study indicated no significant changes in the frequency of sluggish and non-motile sperms of rats in the control group compared to all treatment groups and showed statistically significant differences between the relative frequency of progressive sperms in control group rats compared to all treatment groups except groups 2 and 7. There were no statically significant changes between groups with different time and distance. Sperm count and morphology were not affected by microwave emitted from hair dryer. Conclusion: Exposure to the hair dryer microwaves affects sperm motility, which is amongst the key parameters determining the chance of conceiving.

Logistic Regression Analysis Factors Affecting Sperm Motility and Abnormal Sperm Morphology in Boars.

Simple SummaryReduced sperm motility and morphological abnormalities have significant negative impacts on conception rates in sows and are important indexes of semen. The identification of factors that influence sperm motility and morphology will improve boar fertility in commercial herds. On the basis of analysis of 5042 ejaculates from 385 boars using a logistic regression model, we found that serum Cu excess, serum Fe deficiency, and Pb presence in seminal plasma were risk factors for poor semen quality in boars. More importantly, the presence of seminal plasma Pb had more serious effect on the probability of abnormal sperm morphology than serum Cu excess and serum Fe deficiency. In addition, Yorkshire and Landrace boars had higher sperm motility and lower abnormal sperm morphology than Duroc boars. The difference in serum and seminal plasma elements among boars with different semen qualities may become a guide for regulating these elements used in boar diet. The highly predictive values of serum Cu, Fe, and seminal plasma Pb could be used in the future as an additional tool in semen quality evaluation.Logistic regression models, including variables of boar breed, age, serum, and seminal plasma elements, were used to identify the influencing factors of sperm motility and morphology in this study. Sperm motility degree was classified as grade 0: ≤85% and grade 1: >85%. Abnormal sperm morphology was classified as grade 0: ≤10%, grade 1: 10–20%, and grade 2: >20%. Element concentration of 385 boars was detected by inductively coupled plasma mass spectrometry. Results showed that boars with serum Cu ≥ 2.5 mg/L had lower sperm motility (odds ratio (OR): 0.496; 95% confidence interval (CI): 0.285–0.864) and higher abnormal sperm morphology (OR: 2.003; 95% CI: 1.189–3.376) than those with serum Cu ≤ 2.0 mg/L. Boars with serum Fe ≥ 1.5 mg/L had lower abnormal sperm morphology than those with serum Fe ≤ 1.0 mg/L (OR: 0.463; 95% CI: 0.255–0.842). The presence of Pb in seminal plasma increased abnormal sperm morphology. The probability of abnormal sperm morphology >20% from boars with seminal plasma Pb increased with a range of 5.78–15.30% than that from boars without seminal plasma Pb among three breeds. In conclusion, serum Cu excess, serum Fe deficiency, and seminal plasma Pb are risk factors for poor semen quality in boars.

Three Bacteriocin Peptides from a Lactic Acid Bacterium Weissella confusa MBF8-1 with Spermicidal Activity.

The development of antibiotic resistance amongst bacterial pathogens and a population explosion, e.g. in countries such as Indonesia, are two issues the world is facing today. These issues have stimulated interest in the development of new antimicrobial therapeutic agents and contraceptive strategies, such as novel spermicides. Bacteriocins, which are bacterially-derived antimicrobial peptides, may fulfill some of the criteria for these new agents. Weissella confusa MBF8-1, originally isolated from a homemade soy product, exhibits antibacterial activity that was subsequently found to be plasmid-encoded, presumably by three peptides Bac1, Bac2 and Bac3. In the present study, we tested cell-free MBF8-1 bacteriocin preparations and chemically-synthesized versions of Bac1, Bac2 and Bac3 peptides for (i) its antibacterial activity against the indicator bacterium Leuconostoc mesenteroides and (ii) its ability to affect the motility of spermatozoa. Nisin, a known lantibiotic bacteriocin, was used as the control. Here, we demonstrate that synthetic Bac1, in combination with synthetic Bac2, was sufficient to inhibit the growth of L. mesenteroides and affect sperm motility. However, the presence of all three synthetic peptides, s-Bac1, s-Bac2 and s-Bac3, was required for full potency. In summary, the bacteriocin-like peptides of W. confusa MBF8-1 have the potential to be developed as a narrow-spectrum antimicrobial agent and a novel spermicidal agent.

Dietary Supplementation with Sesame Seed to Improve Semen Quality of Ho Cocks

High levels of polyunsaturated fatty acids in chicken spermatozoa make them susceptible to lipoperoxidation and reduce their fertility. This study was conducted to assess the effect of sesame seed suplementation in the diet on the semen quality of Ho cocks. Eighteen 13-14 month-old cocks were randomly divided into three groups and were assigned to one of the following treatments: 0% SS (control), 5% SS, or 7% sesame seeds per kg of diet for ten consecutive weeks after a two-week adaptation period. Semen characteristics were evaluated once a week. In the 7% sesame seed treatment group, seminal traits including semen ejaculate volume (1.02mL), sperm concentration (3.68 x 109 sperm), and abnormal spermatozoa (10.51% were improved (P &lt;0.05) compared to the control group (0.82mL, 2.81 x 109 sperm, and 11.04% for semen ejaculate volume, sperm concentration, and abnormal spermatozoa, respectively). Supplementation with sesame seeds did not significantly affect sperm motility, mass movement, or semen pH. Our results demonstrate that sesame seed supplementation at 7% successfully improved the ejaculate volume, sperm concentration, and normal spermatozoa percentage of Ho cocks.