Nutritional modulation of the gut-reproductive axis: multi-strain probiotic blend on oxidative and seminal parameters in healthy male dogs.

The aim of this work was to evaluate the effects of the supplementation of two extra-virgin olive oils (EVOO) having different polyphenols content, on canine spermatozoa kinetic parameters and seminal plasma oxidative status. The study was conducted on 12 clinically healthy dogs of different breeds (2-7years, 5-48kg of body weight) divided into two groups: an experimental group supplemented with EVOO (Coratina cultivar) high in polyphenols (H-P) and a control group fed EVOO (Cima di Bitonto cultivar) low in polyphenols (L-P). The oil was daily administered per os (1ml/3kg BW) before meal. Semen collection was made twice at 15days distance (D01 and D02 ) and then at 30 (D30), 60 (D60) and 90 (D90) days. Semen concentration and kinetic parameters were measured using computer-assisted sperm analysis (CASA) system to evaluate: sperm total count, sperm motile (MOT%), progressive motility (PROGR%) and its fractions, straight-line velocity (VSL, μm/s), curvilinear velocity (VCL, μm/s), average path velocity (VAP, μm/s), amplitude of lateral head displacement (ALH, μm), beat cross frequency (BCF, Hz), straightness (STR%) and linearity (LIN%). On seminal plasma, reactive oxygen species (ROS) and biological antioxidant potential (BAP) were tested. From findings, no differences were found for sperm MOT, VSL, VCL, VAP, ALH, BCF, STR, LIN and BAP. A gradual enhancement of PROGR% was observed in H-P group (p<.01). The ROS levels were higher in dogs H-P compared to the other group (p<.05). In conclusion, our results highlight the positive effects of EVOO polyphenols on sperm PROGR% in healthy dogs.

Computer-assisted sperm class analyser (CASA) analysis of avian semen following cryopreservation indicates that their semen motility and viability parameters become compromised, due in part to oxidative stress. To mimic these observations we have treated cockerel semen with an oxidative stress inducing agent, namely hydrogen peroxide (H2O2) and monitored the motility, kinematic and viability parameters over time. Briefly, five healthy and fertile South African Venda cockerels were selected and their semen was collected using the abdominal massage technique. The semen was then treated with H2O2 at 0 &microM, 5 &microM, 50 &microM and 200 &microM concentrations for 0, 3, 16 and 24 hrs. The semen motility, kinematic and viability parameters were then determined using the CASA system while the viability was determined using the SYBR-14/PI staining. The Pearson's correlation coefficient was determined to test the relationships between the levels of induced oxidative stress, period of exposure to oxidative stress inducing agent and the motility plus kinematic parameters. Our data revealed that in raw cockerel semen, there was high and positive correlations between total motility (TM), progressive motility (PM), rapid (RAP), curvilinear velocity (VCL), straight line velocity (VSL) and average path velocity (VAP) while the kinematic parameters LIN, STR, WOB, ALH and BCF had low or negative correlations with them. Furthermore, TM, PM, RAP, VCL and VSL remained highly and positively correlated with the induced oxidative stress and also, linearity (LIN), straightness (STR), wobble (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) remained negatively correlated with the induced oxidative stress, after 3 hrs. After 24 hrs, TM, PM, RAP, VCL, VSL, VAP and ALH, became negatively correlated with the induced oxidative stress while LIN, STR, WOB and BCF became positively correlated with the induced oxidative stress. Conversely, when the H2O2 concentration used was correlated with motility and kinematic parameters over time, TM, PM, RAP, VCL, VSL, VAP became negatively correlated with oxidative stress while LIN, STR, WOB, ALH and BCF show negative or low correlations with the induced oxidative stress. This data indicates that LIN, STR, WOB, BCF and to some extend ALH, reveal the least correlations with the induced oxidative stress under persistent oxidative stress conditions in cockerel semen. In conclusion, cockerel semen, like buck semen, does not easily succumb to oxidative stress since the raw semen correlations of CASA analysed parameters are comparable to these observed after 3 hrs of H2O2 treatment. In addition, the oxidative stress levels tolerated by cockerel semen should not 5 &microM H2O2 oxidative stress levels. Lastly, lack of correlation between LIN, STR, WOB, BCF and ALH and induced oxidative stress can be used in cockerel semen to show intolerable cryopreservation conditions.

Effects of dietary alpha-lipoic acid supplementation on the seminal parameters and fertility potential in aging broiler breeder roosters

Rat epididymal sperm motion changes induced by ethylene glycol monoethyl ether, sulfasalazine, and 2,5-hexandione

This collaborative study was conducted to determine the utility and sensitivity of nine sperm motion parameters generated by a Hamilton-Thorne Sperm Analyzer (HTM-IVOS) for detecting adverse effects of chemicals on sperm motion in rats. The efficacy of sperm motion parameters was investigated using nine reproductive toxicants: adriamycin, alpha-chlorohydrin (3 different studies were carried out), dinoseb, ethylene glycol monoethyl ether, 2,5-hexanedione, sulfasalazine, trimethyl phosphate, and ornidazole. The percentage of motile sperm (% motile sperm), the only parameter expressing the status of semen containing non-motile sperm, detected adverse effects on sperm motion in 9 out of 10 studies. However, weak effects on sperm motion were not detected by this parameter in 4 out of 7 studies in which sperm motion disorders were noted at medium or low dosages. The percentage of progressively motile sperm (% progressive sperm) and the sperm velocity parameters (average path velocity, straight line velocity, and curvilinear velocity) detected adverse effects on sperm motion in all studies. In 7 studies which noted sperm motion disorders at medium or low dosages, weak effects on sperm motion were detected by the % progressive sperm in 5 studies and by the sperm velocity parameters in 6 studies. In 10 studies, amplitude of lateral head displacement (ALH) did not detect adverse effects on sperm motion in 4 studies, and beat cross frequency (BCF) failed to detect adverse effects on sperm motion in 3 studies. Because ALH and BCF show the swimming pattern of spermatozoa as head movement, the characteristics of these parameters are different from the % progressive sperm and the sperm velocity parameters. Straightness (STR) and linearity (LIN), which are secondary parameters calculated from sperm velocity parameters, could not detect adverse effects on sperm motion when the sperm velocity parameters did not detect adverse effects. On the basis of these results, we concluded that the % progressive sperm and sperm velocity parameters are useful and sensitive indicators for detecting adverse effects on sperm motion. However, in the % progressive sperm, setting up a suitable threshold of VAP and/or STR is important to gain further sensitivity for detecting adverse effects on sperm motion. The % motile sperm is useful for assessment of sperm motion disorder, and ALH and BCF are useful for evaluating the swimming pattern of sperm. STR and LIN are not very useful for detecting adverse effects on sperm motion.

Suspensions of seminal plasma-free human spermatozoa were prepared by swim-up from semen and studied using high magnification videomicrography after incubation under capacitating conditions for 1.5-2 h. Three subpopulations of capacitating spermatozoa showing different patterns of motility could be distinguished visually: forward progressive, transition phase, and hyperactivated motility. The purpose of this study was not to determine the relative proportions of spermatozoa in these three categories but to describe their movement characteristics. Manual track plotting and analysis allowed value derivation for the curvilinear, average path and straight-line velocities (VCL, VAP, and VSL respectively); for the three progression ratios of linearity (LIN = VSL divided by VCL X 100), straightness (STR = VSL divided by VAP X 100), and wobble (WOB = VAP divided by VCL X 100); and also for the amplitude of lateral head displacement (ALH) and the beat/cross frequency (BCF). Algorithms produced from these motion characteristics allowed distinctions to be made between cell motility patterns. Spermatozoa with straight-line velocity (VSL) greater than or equal to 40 microns/s, linearity (LIN) greater than or equal to 60% and amplitude of lateral head displacement (ALH) less than 5 microns were FP or non-hyperactivated. Tracks with curvilinear velocity (VCL) greater than or equal to 100 microns/s, linearity (LIN) less than 60% and amplitude of lateral head displacement (ALH) greater than or equal to 5 microns showed concomitants of hyperactivation. Classical hyperactivated tracks also showed straightness (STR) less than 60% and straight-line velocity (VSL) less than 30 microns/s.

This study evaluated the impact of pre-freezing semen dilution rate and dimethyl acetamide (DMA) concentration on the post-thaw motility and fertility of cryopreserved rooster sperm. Rooster ejaculates were diluted with a standard EK extender to achieve low (LSC; 1 × 10⁹ sperm/mL) and high (HSC; 2 × 10⁹ sperm/mL) sperm concentrations. Each dilution group was further treated with three DMA concentrations (3%, 6%, or 9%) before cryopreservation. Post-thaw sperm motility traits were obtained by computer-assisted sperm analysis (CASA), and fertility features were evaluated through artificial insemination in hens. The current results showed that HSC significantly improved total motility (TM), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF), but reduced linearity (LIN) and straightness (STR) compared to LSC. DMA concentration had a quadratic effect on motility, with 6% yielding the highest progressive motility (PM), straight line velocity (VSL), and BCF. Fertility outcomes revealed that HSC resulted in higher fertilization rates, while neither DMA concentrations nor their interaction with dilution rates exerted significant effects on fertility traits. VCL, ALH, and BCF showed positive correlations with pipping-chicks rates, whereas STR, LIN, and WOB displayed negative correlations. These findings underscore the critical interplay between dilution rate and cryoprotectant concentration and provide practical guidance for developing more reliable cryopreservation protocols that can be applied under field conditions to enhance fertility management in poultry production.

This study evaluated the impact of pre-freezing semen dilution rate and dimethyl acetamide (DMA) concentration on the post-thaw motility and fertility of cryopreserved rooster sperm. Rooster ejaculates were diluted with a standard EK extender to achieve low (LSC; 1 × 10⁹ sperm/mL) and high (HSC; 2 × 10⁹ sperm/mL) sperm concentrations. Each dilution group was further treated with three DMA concentrations (3%, 6%, or 9%) before cryopreservation. Post-thaw sperm motility traits were obtained by computer-assisted sperm analysis (CASA), and fertility features were evaluated through artificial insemination in hens. The current results showed that HSC significantly improved total motility (TM), curvilinear velocity (VCL), amplitude of lateral head displacement (ALH), and beat cross frequency (BCF), but reduced linearity (LIN) and straightness (STR) compared to LSC. DMA concentration had a quadratic effect on motility, with 6% yielding the highest progressive motility (PM), straight line velocity (VSL), and BCF. Fertility outcomes revealed that HSC resulted in higher fertilization rates, while neither DMA concentrations nor their interaction with dilution rates exerted significant effects on fertility traits. VCL, ALH, and BCF showed positive correlations with pipping-chicks rates, whereas STR, LIN, and WOB displayed negative correlations. These findings underscore the critical interplay between dilution rate and cryoprotectant concentration and provide practical guidance for developing more reliable cryopreservation protocols that can be applied under field conditions to enhance fertility management in poultry production.

The aim of this study was to use computer-assisted sperm analysis (CASA) to examine changes in motion parameters of rat spermatozoa incubated under culture conditions that support IVF. Rat cauda epididymal spermatozoa were evaluated in six replicate experiments, at 0 and 4h of incubation. CASA was conducted at 60 Hz on digital 1s tracks ( approximately 100 spermatozoa/rat). Mean values of CASA parameters that describe the vigour of spermatozoa [curvilinear velocity (VCL), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF)] increased, while those indicating progressiveness [straight line velocity (VSL), linearity (LIN) and straightness (STR)] decreased between 0 and 4 h. Visual inspection of sperm tracks after 4 h of incubation revealed classical hyperactivation patterns. Bivariate models were evaluated to objectively define the subpopulation of hyperactivated (HA) spermatozoa. Of all models considered, ALH and LIN, VCL and LIN, BCF and LIN, VCL and BCF, and VCL and ALH showed significant changes in the percentage of HA spermatozoa after the 4 h incubation period. The efficacy of detecting HA spermatozoa was evaluated using sperm tracks that were visually classified as HA or progressive. VCL and LIN provided the most accurate prediction of HA spermatozoa. It was concluded that analysis of CASA data using bivariate models could be used to detect and monitor hyperactivation in rat spermatozoa.

Effects of melatonin implants on the reproductive performance and endocrine function of camel (Camelus dromedarius) bulls during the non-breeding and subsequent breeding seasons

At present, frozen-thawed semen is used extensively for artificial insemination (AI) in Ethiopia. However, subjective semen quality assessment is still practiced by the semen processing organizations of the country. In this study, motility and motion characteristics of spermatozoa were assessed by using integrated semen analysis system (ISAS) to diagnose breed differences in quality of semen. Semen was collected from 14 breeding bulls (Boran = 4, Crosses of 75 % Holstein Frisian ËŸ 25% Boran = 4 and Holstein Frisian = 6). After initial subjective assessment, the semen was evaluated at fresh, chilled and frozen stages for various sperm motion characteristics using integrated semen analysis system (ISAS&reg; v1, Proiser, Spain). PH, volume and morphological defects were differed significantly (P &lt; 0.05) among breeds. Significantly higher (P &lt; 0.05) motility percentage (82.5%) was identified in Boran fresh semen. In contrast spermatozoa motion characteristics: Medium, medium progressive and slow types were significantly higher (P &lt; 0.05) in crosses. Individual motility percentage and spermatozoa motion types (medium and slow) were significantly low (P &lt; 0.05) in Boran chilled semen. In line with chilled semen significantly high motility percentage (42.9%) for frozen semen was observed in HF. The sperm kinematic parameters: Average path velocity (VAP, &micro;m/s), straight line velocity (VSL, &micro;m/s), curvilinear velocity (VCL, &micro;m/s), amplitude of lateral head displacement (ALH, &micro;m), beat cross frequency (BCF, Hz) and straightness (STR) percentage were differed significantly (P &lt; 0.05) among the three breeds at all stages of semen production. The smallest kinematic values of all parameters except for LIN, STR and WOB of fresh semen were recorded in cross breed bulls at all stages of production. On the other hand significantly higher (P &lt; 0.05) values for all the parameters (VCL, VAP, VSL, ALH, BCF, LIN, STR and WOB) of frozen semen were recorded in Boran breed. Thus it is possible to conclude that breed has influence on motility and motion characteristics of bull spermatozoa at different stages of semen production. Key words: Boran, Integrated Semen Analysis System (ISAS), kinematic parameters.

Study question How to construct a machine learning-based assessment system for sperm motility and pick up the superior sperm with high motility accurately and efficiently? Summary answer Machine learning algorithms can be trained to build a grading model for the classification and grading of sperm motility. What is known already There are many sperm quality parameters for male fertility evaluation such as sperm concentration, viability, morphology, pH, and color of the semen. But, many studies agree that motility is the main parameter for sperm quality evaluation. Currently, there are mainly two methods for sperm motility determination. The traditional way relies on manual observation under a microscope, but this is subjective and time-consuming. Though there are some tracking errors in computer assisted sperm analysis (CASA) system, which help to acquire more motion parameters such as curvilinear velocity (VCL), straight line velocity (VSL) and average path velocity (VAP). Study design, size, duration 3 000 sperm samples from clinical laboratory were divided to asthenospermia (AS) group and healthy control (HC) group based on their progressive motility. Sperm motility parameters were measured by CASA system. Participants/materials, setting, methods Nine variables of sperm motility including VCL, VSL, VAP, amplitude of lateral head displacement (ALH), mean angular displacement (MAD), linearity (LIN), straightness (STR), beat-cross frequency (BCF), fractal dimension (D) were collected for ML model training. The R software was used to build several classifiers on the samples investigated, including Random Forest, Adaptive Boosting (Ad Boost), Gradient Boosting, Support Vector Machine map (SVM), kNN (k-Nearest Neighbours), and Naïve Bayes. Main results and the role of chance Motility parameters differ between the two groups with a significant difference in VAP, VSL, VCL, and ALH. In order to classify the heterogeneous dataset in an unbiased manner, we performed a cross-validation procedure of training learning algorithms using six folds (equal parts) of the dataset, each time using a separate fold as the test dataset and the remaining folds as the training dataset. We initially examined whether the classifier could be reliably predicted using a subset of the sperm characterization’s 9 parameters (VCL, VSL, VAP, ALH, MAD, LIN, STR, BCF, D). We purposefully excluded additional characteristics that may be linearly related to increasing motility. We found four learning algorithms to have high accuracy, recall, precision, and F1 score. The support Vector Machine map performed best in classifying the sperm with high motility. Limitations, reasons for caution There may be some bias in data for sperm motility parameters. Specifically, different data collected from different laboratories or acquired from different equipment which may lead to a degradation in the performance of the machine learning model. Wider implications of the findings The analysis of large amounts of case data by machine learning algorithms can help clinician better understand the sperm motility characteristics of different patients so that they can develop a personalized treatment plan for each patient. Trial registration number not applicable

Impact of age, clinical conditions, and lifestyle on routine semen parameters and sperm kinematics

The aim of the study was to determine the sperm motility parameters in wild Atlantic salmon and sea trout to define criteria important for selection of milt for controlled fertilisation. Parameters for these species were determined in the fish migrating into north-western rivers of Poland at spawning time. Eight motility parameters percentage of motile sperm (MOT), curvilinear velocity (VCL), average path velocity (VAP), straight line velocity (VSL), linearity (LIN), straightness (STR), amplitude of lateral head displacement (ALH), beat cross frequency (BCF) and motility duration were subjected to computer-assisted sperm analysis (CASA). Milt of most individuals studied representing both salmon and trout showed spermatozoa density of 12–22 × 109 ml−1 and a high percentage of motile sperm (>70%). In general, spermatozoa swim progressively with slightly curved trajectories (mean STR = 70%, LIN = 65%) and velocity VCL of 180 μm s−1 (salmon) and 190 μm s−1 (trout), at 10 s post-activation. Such sperm is easily accessible in the wild populations of salmon and sea trout and is recommended for use in reproduction trials. The spermatozoa of sea trout seem to show a greater tendency to follow curvilinear trajectories than those of salmon, both in the beginning and the final phase of motion. In the first phase of motility, the values and time dependencies of the motility parameters were similar in both species. In the end phase of movement differences in LIN and BCF time dependencies were found in the samples representing the two species. In salmon the linearity and beat cross frequency remained stable in this phase, contrary to the patterns in sea trout for which LIN decreased while BCF increased in the end period of movement. Durations of movement were similar in both species (ranges of 20–40 s).

Effect of resveratrol on post-thaw motility, kinematics, structural parameters and antioxidant/oxidant status of Kamori buck spermatozoa.