Total Number Of Spermatozoa Research Articles

Impact of environmental variables on some reproductive metrics of white-lipped peccaries (Tayassu pecari) raised in Atlantic Forest of Brazil

Knowledge on male reproductive physiology is essential for the development of effective conservation strategies. This study investigated the influence of environmental variables on certain reproductive metrics in white-lipped peccaries (Tayassu pecari) raised in the Atlantic Forest. After anesthetization, testicular and cauda epididymis biometry were evaluated in nine adult male individuals subjected to electroejaculation. Semen was evaluated for volume, pH, concentration, total number of sperm, sperm morphology, membrane integrity, and kinematic parameters. Concurrently, environmental variables were collected from the day before, for the previous 14 days (estimated for sperm maturation in epididymis), and the period of 51–55 days (corresponding to the spermatogenic cycle) before semen collection. Overall, it was observed that rainfall is the most important environmental variable influencing the reproductive parameters of white-lipped peccaries, being positively correlated with the amplitude of lateral sperm head displacement (ρ = 0.62, P < 0.05) and the appearance of proximal cytoplasmic droplets in sperm (ρ = 0.62, P < 0.05). In addition, the testicular biometry of the species is influenced by the set of environmental variables of air temperature, rainfall, and relative humidity (ρ ≥ 0.60, P < 0.05). On the other hand, epididymal biometric data showed numerous correlations between cauda epididymis metrics and sperm parameters (ρ = 0.68, P < 0.05). This information will be useful to improving conservation strategies for these animals, contributing to their management in captivity and to reintroduction programs, especially in the Atlantic Forest where the species is declining.

Sperm quality metrics were improved by a biomimetic microfluidic selection platform compared to swim-up methods

Sperm selection is an essential component of all assisted reproductive treatments (ARTs) and is by far the most neglected step in the ART workflow in regard to technological innovation. Conventional sperm selection methodologies typically produce a higher total number of sperm with variable motilities, morphologies, and levels of DNA integrity. Gold-standard techniques, including density gradient centrifugation (DGC) and swim-up (SU), have been shown to induce DNA fragmentation through introducing reactive oxygen species (ROS) during centrifugation. Here, we demonstrate a 3D printed, biologically inspired microfluidic sperm selection device (MSSP) that utilizes multiple methods to simulate a sperms journey toward selection. Sperm are first selected based on their motility and boundary-following behavior and then on their expression of apoptotic markers, yielding over 68% more motile sperm than that of previously reported methods with a lower incidence of DNA fragmentation and apoptosis. Sperm from the MSSP also demonstrated higher motile sperm recovery after cryopreservation than that of SU or neat semen. Experiments were conducted side-by-side against conventional SU methods using human semen (n = 33) and showed over an 85% improvement in DNA integrity with an average 90% reduction in sperm apoptosis. These results that the platform is easy-to-use for sperm selection and mimics the biological function of the female reproductive tract during conception.

Результаты мультицентровой наблюдательной программы по изучению эффективности и безопасности многодозового менотропина у пациентов с мужским бесплодием.

Introduction. Currently, menotropins are widely used to treat male infertility, both to stimulate spermatogenesis and to compensate for hormonal imbalance. But the actual practice of using menotropins may differ from the recommendations given by the manufacturer. Therefore, observational programs are particularly important, which allow assessing the effectiveness of a drug in routine practice. Purpose. To establish prognostic criteria for response to short and stadard course of multidose menotropin therapy in men with pathospermia. Materials and methods. The study was designed as a multicentre, prospective, observational cohort study. The study included 1120 men with infertility enrolled between January 2020 and April 2021. The study included men with the absence of pregnancy during 12 months of regular unprotected sexual activity. The average duration of infertility was 20.6 months. As a treatment, men received multidose menotropin at a dosage of 75 IU, 112.5 IU or 150 IU every other day with hCG 2000 IU 2-3 times a week. Sperm parameters and hormones (total testosterone, Sex hormone-binding globulin - SHBG, luteinizing hormone - LH, follicle stimulating 12 months - FSH, Inhibin B, estradiol) were evaluated before treatment and for 12 months. Results. When comparing the parameters of the spermogram, it was noted that without differentiation according to the prescribed dose, after 3 months of administration, the concentration of spermatozoa increased by 32.6%, the total number of spermatozoa in the ejaculate increased by 32.3%, progressive mobility increased by 20.7%, the proportion of morphologically normal forms increased increased by 34.8%. The level of total testosterone after 3 months of use of multidose menotropin increased by 33.6%. The levels of inhibin B and estradiol did not change significantly. When using multi-dose menotropin for 12 months, sperm concentration increased by 88.5%, the total number of spermatozoa in the ejaculate increased by 55.3%, progressive mobility increased by 42%, the proportion of morphologically normal forms increased by 64.3%. Conclusions. We found the increase of the effectiveness of menotropins with an increase in the duration of their use and an increase in dosage.

Bladder Neck Obstruction: Experience and Management in a Sperm Bank.

Primary Bladder Neck Obstruction (PBNO) management provides medical and surgical treatment, such as transurethral incisions that can lead to retrograde ejaculation. The aim of this study was to investigate the maintenance of anterograde ejaculation and semen quality before and after this surgical procedure. A retrospective evaluation was carried out between 2011 and 2020. A total of 73 patients diagnosed with PBNO were recruited. Ejaculatory function, semen quality, and the fertility of recruited subjects were evaluated. Semen parameters-Baseline, 8.2% of patients were oligozoospermic and 12.3% had a semen volume below the WHO 2010 fifth percentile. Post-surgery, 20% of patients were oligozoospermic. We detected a significant decrease in total sperm number, a significant increase in the number of abnormal forms, and a reduction in the leukocyte concentration. Ejaculatory function-A total of 7.7% of patients reported anejaculation after transurethral incision of the bladder neck. Fertility-9.2% of the patients already had children before surgery; 13.8% had naturally conceived children in the years following surgery; 76.9% had no desire for paternity at the time. Our data have important implications for sperm bank management. The alterations in semen parameters and the risk of anejaculation suggest that the use of sperm cryopreservation before surgery for PBNO should be encouraged.

The Loss-Function of KNL1 Causes Oligospermia and Asthenospermia in Mice by Affecting the Assembly and Separation of the Spindle through Flow Cytometry and Immunofluorescence

KNL1 (kinetochore scaffold 1) has attracted much attention as one of the assembly elements of the outer kinetochore, and the functions of its different domains have been gradually revealed, most of which are associated with cancers, but few links have been made between KNL1 and male fertility. Here, we first linked KNL1 to male reproductive health and the loss-function of KNL1 resulted in oligospermia and asthenospermia in mice (an 86.5% decrease in total sperm number and an 82.4% increase in static sperm number, respectively) through CASA (computer-aided sperm analysis). Moreover, we introduced an ingenious method to pinpoint the abnormal stage in the spermatogenic cycle using flow cytometry combined with immunofluorescence. Results showed that 49.5% haploid sperm was reduced and 53.2% diploid sperm was increased after the function of KNL1 was lost. Spermatocytes arrest was identified at the meiotic prophase I of spermatogenesis, which was induced by the abnormal assembly and separation of the spindle. In conclusion, we established an association between KNL1 and male fertility, providing a guide for future genetic counseling regarding oligospermia and asthenospermia, and a powerful method for further exploring spermatogenic dysfunction by utilizing flow cytometry and immunofluorescence.

Semen Quality of the First and Second Ejaculates Collected from Breeding Inactive Stallions after Cooling and Freezing

This study aimed to assess the semen quality after the cooling and freezing of the first and second ejaculates of the season, which were collected 1 h apart. After collection (n = 40 ejaculates), the gel-free semen volume, concentration, total number of sperm, and sperm morphology were determined. An aliquot of each ejaculate was extended and cooled for 48 h; a second aliquot was cushion-centrifuged and cooled for 48 h; and a third aliquot was processed and then frozen. The total motility (TM) and progressive motility (PM), plasma membrane integrity (PMI), and high mitochondrial membrane potential (HMMP) were assessed pre-(0 h), 24 h, and 48 h post-cooling and before and after freezing. The second ejaculate had a lower gel-free semen volume (p = 0.026). The sperm concentration was greater in the first than in the second ejaculate (p < 0.001). The sperm morphology was similar between the ejaculates (p > 0.05). Cushion-centrifugation prevented a reduction in the TM, PM, and PMI over time (p < 0.05). The TM, PM, and PMI decreased after freezing but not between the ejaculates (p > 0.05). The first and second ejaculates of the season, which were collected 1 h apart, varied in quantity but not in quality after cooling and freezing.

Ram Semen Cryopreservation for Portuguese Native Breeds: Season and Breed Effects on Semen Quality Variation

The semen quality is one of the determinant factors of ram semen cryopreservation. The present retrospective study aimed to characterize the seasonal ram pattern during the year for ten Portuguese local sheep breeds, hypothesizing that the breed and season had low effects on the main spermatozoa traits. A total of 1471 ejaculates were used and evaluated (fresh semen) from 85 rams between 2004 and 2020 and re-evaluated after thawing (thawed semen). The effect of breed, season, and sperm cryopreservation on nine semen traits were evaluated. The volume per ejaculate, spermatozoa (SPZ) concentration, and total number of SPZ per ejaculate, were affected by breed (p < 0.001) but not by season (p > 0.05). As expected, the semen processing was the most significant (p < 0.001) factor of variation on seminal parameters. Moreover, breed and interactions between breed × semen processing, modulated the response of alive SPZ, abnormal morphology, head, and intermediate piece defects. In fresh semen, season only affected the intermediate piece defects due to the highest percentage observed between February and April period in some breeds. Overall, and despite the mentioned particularities, there were similarities among the ten local breeds. We also concluded that the seasonal effect on ejaculate and SPZ traits is not significant in our region. These local ram breeds have low seasonality and can be employed in natural mating as well as semen donors for cryopreservation and assisted reproductive biotechnologies during the whole year at our latitude.

A decade of WHO 2010: total sperm number temporal trend and role of lifestyle factors.

After a huge decline in sperm concentration between 1938 and 1991 was reported, many researchers investigated the possibility of a worsening of human sperm quality. Despite massive efforts, published evidence is still controversial. Similarly, the role of lifestyle factors on semen parameters is debated. We conducted a monocentric Italian study to evaluate the total sperm number trend over the last 10 years (from 2010 to 2019). Additionally, we evaluated the association between lifestyle factors and total sperm number in order to identify possible damaging factors. We performed a retrospective study analyzing subjects aged 18-55 years who had their semen analyzed between 2010 and 2019. A total of 3329 subjects were included: 1655 subjects referred to our department (Department of Experimental Medicine, Sapienza University of Rome, Roma, Italy) for idiopathic infertility and 1674 subjects referred for preconceptional or andrological screening with no confirmed andrological diseases. Semen samples were examined according to World Health Organization (WHO) 2010 criteria by two seminologists with the same training and the same equipment. For statistical evaluations, only total sperm number (×10 6 per ejaculate) was taken into consideration. We detected no significant changes in mean total sperm number during the last decade, in either the entire population or the two subgroups (infertile group and control group). In a multivariate analysis total sperm number was significantly associated with the history of infertility, body mass index (BMI) and cigarette smoking. Our results suggest that infertile men are "vulnerable" subjects, particularly susceptible to several negative factors, many of which still remain unknown. Our study highlights the need for studies addressing men's lifestyle in order to find and reduce deleterious agents.

Sulfur dioxide may predominate in the adverse effects of ambient air pollutants on semen quality among the general population in Hefei, China

Previous studies have reported potential adverse effects of exposure to ambient air pollutants on semen quality in infertile men, but studies on the general population have been limited and inconsistent, and the pollutants that play a major role remain unclear. This study aimed to explore the potential association between exposure to six air pollutants (PM2.5, PM10, NO2, SO2, O3 and CO) during different sperm development periods and semen quality among the general population, and to explore the interaction between different air pollutant exposures. We included 1515 semen samples collected from the Human Sperm Bank. We improved individuals' exposure level estimation by combining inverse distance weighting (IDW) interpolation with satellite remote sensing data. Multivariate linear regression models, restricted cubic spline functions and double-pollutant models were used to assess the relationship between exposure to six air pollutants and sperm volume, concentration, total sperm number and sperm motility. A negative association was found between SO2 exposure and progressive motility and total motility during 0–90 lag days and 70–90 lag days, and SO2 exposure during 10–14 lag days adversely affected sperm concentration and total sperm number. Sensitive analyses for qualified sperm donors and the double-pollutant models obtained similar results. Additionally, there were nonlinear relationships between exposure to PM, NO2, O3, CO and a few semen parameters, with NO2 and O3 exposure above the threshold showing negative correlations with total motility and progressive motility, respectively. Our study suggested that SO2 may play a dominant role in the adverse effects of ambient air pollutants on semen quality in the general population by decreasing sperm motility, sperm concentration and total sperm number. Also, even SO2 exposure lower than the recommended standards of the World Health Organization (WHO) could still cause male reproductive toxicity, which deserves attention.

Analysis of artificial insemination center management factors that contribute to sperm parameters and boar longevity with a major focus on PGF2α treatment.

Artificial insemination (AI) centers' economic profitability is directly impacted by the amount of high-quality sperm doses. Many internal and external factors contribute to the quality of ejaculates. To enhance the libido and to reduce the time that is required to train boars for semen collection, prostaglandin products are used routinely at AI centers. The objectives of this study were to analyze sperm parameters of 40,765 ejaculates and the survival of 406 PGF2α-treated and 417 untreated AI boars of five breeds (Duroc [DU], Large White [LW], Landrace [LR], Pietrain [PI], and Large White Sire Line [LS]). Data were assessed in one AI center in Switzerland from 2018 to 2022 and sperm parameters were determined by computer-assisted sperm analysis immediately after semen collection. For further calculations, the ejaculates were divided into four groups depending on the boar's age at semen collection day: 7 to 11, 12 to 18, 19 to 24, and 25 to 87 mo. Along with the boar age, breed, season, and semen collection frequency even the treatment with PGF2α had significant influence on sperm parameters. Overall, 19.5% ejaculates were collected after PGF2α treatment. For every age and breed group, higher ejaculate volume was found for untreated boars than for PGF2α-treated boars (P < 0.001). Higher values for total and progressive sperm motility, total sperm number, and the proportion of morphologically normal sperm were observed in untreated boars across several age and breed groups. Only for sperm concentration, higher values were found for treated boars older than 11 mo (P < 0.001) and the breeds DU, LW, and LR (P < 0.001). The chance of ejaculates being rejected for AI purposes because of low sperm quality was 21% greater for treated boars. There was no difference in survival between treated and untreated boars. The hazard for boar removal increased with decreasing age at first semen collection and decreasing semen collection frequency. The results reveal better sperm quality for untreated boars than for PGF2α-treated boars, which might indicate that the underlying fertility of boars necessitating treatment is inferior compared to boars that can be collected without further treatment. Therefore, AI boar stations that are aiming to reduce PGF2α treatment should pay special attention to high libido and fertility of young prospective AI boars.

Use of ketamine associated with detomidine or xylazine for semen collection with electroejaculation in pampas deer (Ozotoceros bezoarticus)

Semen collection by electroejaculation under general anesthesia is a frequent method used in wild animal species. The study aimed to compare the effectiveness of two anesthetic protocols (ketamine/detomidine (KD) or ketamine/xylazine (KX) for semen collection with electroejaculation in pampas deer (Ozotoceros bezoarticus) males. Also, compared the physiological and hematological changes with the two anesthetic protocols. Seven pampas deer males were anesthetized with both protocol in an overcorssed design, reverting the anesthesia with atipamezole in both protocols. The induction time, recovery time, and the time of the anesthesia procedures did not differ between both anesthetic protocols. The heart rate was greater when animals were anesthetized with KD than when KX was used (55.33 ± 2.7 bpm vs 45.12 ± 2.6 bpm; P = 0.05). Although there was no difference according to the anesthetic protocol, the oximetry values were below those considered normal in other species. When animals were treated with the KD, the red blood cell number, hematocrit, and hemoglobin concentration were greater than with the KX protocol (17.1 ± 1.3 ×106/µL vs 15.9 ± 1.3 ×106/µL; P = 0.006; 47.2 ± 3.4% vs 43.9 ± 3.4%; P = 0.008; and 14.8 ± 0.8 g/dL vs 13.8 ± 0.8 g/dL P = 0.01, respectively). Less number of electrical pulses were required for ejaculation in animals treated with KX than with KD (39.8 ± 2.6 vs 52.4 ± 2.6, P = 0.04). In samples collected from animals treated with KD, the total number of sperm with integral acrosome was lower, but the percentage of sperm with normal morphology was greater than in animals treated with KX (67.8 ± 67.0 × 106 sperm vs 243.1 ± 71.6 × 106, P = 0.03 and 40.0 ± 2.3% vs 30.6 ± 2.7%, P = 0.02; respectively). In conclusion, the use of KX induced less physiological changes, appearing also advantageous in semen quality.

INFLUENCE OF THE DEGREE OF PROGRESSION OF VARICOCELE ON THE REPRODUCTIVE HEALTH OF ADOLESCENTS

The main pathogenetic mechanisms of deterioration of reproductive function in varicocele under conditions of hyperthermia, ischemia and hypoxia of the testicular tissue are considered to be a violation of the endocrine function of the testicles as a result of alteration of Leydig cells, the development of an autoimmune form of infertility due to damage to Sertoli cells and the hematotesticular barrier, damage to the germinal epithelium of the seminiferous tubules can lead to disruption of spermatogenesis. The question of the influence of the degree of progression of varicocele on the severity of testicular dysfunction has not been fully studied. Purpose of the study. To assess the impact of the degree of varicocele progression on the reproductive health of adolescents from 14 to 17 years of age. Materials and methods. Adolescents with II and III degrees of varicocele after laparoscopic varicocelectomy from 14 to 17 years of age once annually performed a determination of the level of antisperm antibodies (ASAT), follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, estradiol in the blood serum. When the subject reached the age of 17 years, a semen analysis was performed. Results and discussion. During the observation period, the levels of all studied hormones, with the exception of estradiol, were within the reference interval, and their dynamics reflects the active puberty period. Monitoring of the ACAT level made it possible to establish its increase, but the values did not exceed diagnostically significant values. The study of the spermogram showed that the values of all parameters are within the reference interval. The results of the study were compared and evaluated depending on the degree of progression of varicocele. There was no statistically significant difference between the subgroups in the levels of hormones and ASAT. In terms of spermogram in patients with III degree of varicocele, statistically significantly lower indicators of ejaculate volume, concentration and total number of spermatozoa were established, in general, no critical violation of spermatogenesis was recorded. Conclusion. The conducted study on the assessment of the influence of the degree of varicocele progression on indicators reflecting the state of the reproductive function of adolescents made it possible to establish that, compared with grade II varicocele, with grade III varicocele, statistically significantly lower values were determined only for the following indicators: spermatozoa concentration (84.96 + 55.68 and 62.80 + 41.60; p≤0.025), total sperm count (221.07 + 171, 89 and 144.85 + 153.93, p≤0.021). At the same time, normozoospermia is most often diagnosed in all adolescents during this period. The frequency of occurrence of such a conclusion was 65.3% of cases with grade II varicocele and 39.21% of cases with grade III varicocele, in adolescents without varicocele — 63.3%. In all other studied parameters, there were no statistically significant differences depending on the degree of varicocele. The indicators of all determined parameters, including spermograms, were within the age reference intervals and their diagnostically significant deviation was not recorded. Based on the data obtained, it can be assumed that the degree of progression of varicocele in adolescents does not adversely affect the endocrine function of the testis, the hematotesticular barrier and spermatogenesis in the age period from 14 to 17 years.

Optimizing the ratios of standardized ileal digestible (SID) methionine plus SID cystine and SID threonine to SID lysine in low-protein diets for working boars

This study aimed to optimize the ratios of standardized ileal digestible (SID) methionine (Met) plus cystine (Cys), and threonine (Thr) to SID lysine (Lys) in low-protein diets for working boars. Forty-eight working Duroc boars were randomly allocated to one of 12 dietary treatments in a 3x4 factorial experimental design in which factor 1 was the ratios of SID Met plus Cys to SID Lys (50, 60, 70%), factor 2 was the ratios of SID Thr to SID Lys (40, 50, 60, 70%). Semen was collected at a 4 days interval for 6 weeks for 10 ejaculates. Semen volume (V), percentage of sperm with progressive motility (A), sperm concentration (C), and the total number of motile sperm per ejaculate (VAC) were measured. The results of the study revealed that the ratios of SID Met plus Cys to SID Lys in the diets affected the C and VAC. Values of C and VAC were highest at the ratios of SID Met plus Cys to SID Lys of 70% and lowest at 50% (P&lt;0.05). Similarly, the ratios of SID Thr to SID Lys affected the C and VAC. Further, the values of C and VAC were highest at the ratio of SID Thr to SID Lys of 60% and lowest at 40% (P&lt;0.05). There was no interaction effect between the two factors. In conclusion, the ratios of SID Met plus Cys to SID Lys of 70% and SID Thr to SID Lys of 60% in a 13.5% CP diet are optimal for working boars.

THE REPRODUCTIVE ABILITY OF MALE RABBITS DEPENDING ON THE INFLUENCE OF PARATYPICAL AND GENOTYPICAL FACTORS

THE REPRODUCTIVE ABILITY OF MALE RABBITS DEPENDING ON THE INFLUENCE OF PARATYPICAL AND GENOTYPICAL FACTORS

A Case-Control Study on the Effects of Plasticizers Exposure on Male Fertility.

Male infertility is a serious concern for public health, and the possible role of exposure to plasticizers such as phthalates and bisphenol A in contributing to the condition is widely debated. We have herein enrolled 155 infertility cases attending an infertility center and 211 controls (fathers of a spontaneously conceived newborn) to investigate this issue. The urinary levels of seven phthalates and BPA were analyzed through HPLC/MS/MS. All data were statistically elaborated considering information about clinical situation, life habits, occupational activity, and, for cases, semen parameters (volume, sperm concentration, total count of spermatozoa, and sperm motility). Results showed significantly higher urinary concentrations for all the phthalates in cases compared to controls, except for monoethylphthalate and BPA. In total, 90.07% of cases had sperm motility lower than the WHO reference value (2010), while 53.69%, 46.31%, and 16.56% had sperm total number, concentration, and volume, respectively, out of the reference range. Regarding the possible source of exposure, the use of scents seems to be a significant source of DEP (diethylphthalate). When considering occupational settings, industrial workers, dental technicians, artisans, and farmers using chemicals showed higher risk (OR = 2.766, 95% CI 1.236-6.185), particularly in relation to DnBP (di-n-butyl phthalate) and DEHP (di-ethyl-hexyl phthalate) exposure. No clear quantitative correlation between specific plasticizers and sperm parameters could be demonstrated but these findings call for future studies about the risks associated with exposure to their mixture.

The effect of aging on semen parameters in normozoospermic men: A cross-sectional study.

Semen parameters change with age and are reported differently worldwide. This retrospective cross-sectional study aimed to investigate the semen quality pattern among aging men and the age thresholds for semen parameters. The records of men who had normal semen parameters from January 2015-June 2020 were retrospectively evaluated for andrological outpatient at Samsun Training and Research hospital and Gazi hospital in Samsun, Turkey. Adult men meeting the inclusion criteria were divided into 3 groups of I) 18-29 yr (n = 629), II) 30-39 yr (n = 775), and III) 40-49 yr (n = 190). Correlations between age and sperm parameters were then analyzed. A total of 1594 men were enrolled in the study. Significant differences were observed in total sperm numbers, total motility rates, progressive motility rates, nonprogressive motility rates, normal morphology rates, mean semen volume, and sperm concentrations. The parameters of total sperm number, progressive motility rate, and normal morphology rate were significantly higher in group I than in the other 2 groups (p 0.001, p 0.001, and p 0.001, respectively) and in group II compared to group III (p = 0.001, p = 0.003, and p 0.001), respectively. Mean semen volume and total motility rate were significantly higher in group I than in the other groups (p = 0.001 and p 0.001, respectively). However, no difference was observed between group II and group III (p = 0.61 and p = 0.04, respectively). Age has a significant impact on semen parameters capable of affecting male fertility, particularly total sperm numbers, the progressive motility rate, and the normal morphology rate.

The level of acute phase blood proteins as a predictive criterion of the cryo resistance of stallion sperm

The indicators of the biochemical analysis of the blood of horses reflect the state of metabolism of the animal and at the same time are potential predictors of the qualitative characteristics of spermatozoa, the freezing of which is one of the most important ways to preserve and further use the genetic material. The aim of this study was to identify correlations between the level of such acute phase blood proteins, as C-reactive protein and fibrinogen, and the qualitative and quantitative characteristics of the native and thawed sperm of stallions. The object of the study was blood and sperm samples from 87 stallions of various breeds, the average age of which was 10.5±0.5 years. To assess the protein status of the blood, in addition to the aforementioned acute phase proteins, the total protein content, the levels of albumin, globulins, and the albumin / globulin ratio were determined. Stallion sperm samples were assessed according to the following indicators: ejaculate volume, concentration, the total number of spermatozoa in the ejaculate, progressive motility and survival at +4°C. When analyzing the correlations between the studied parameters, a statistically significant negative inverse relationship was revealed between the level of fibrinogen in the blood serum and such parameters as the content of progressively motile sperm in native and thawed cryopreserved sperm (r=-0.22, p&lt;0.05 and r=-0.29, p&lt;0.01, respectively), as well as sperm survival in native and thawed sperm (r=-0.48, p&lt;0.001 and r=-0.22, p&lt;0.05, respectively).There was no statistically significant relationship between the level of C-reactive protein and the studied characteristics of the spermogram. The revealed negative correlation between the level of fibrinogen and the characteristics of the stallions’ sperm can be considered as one of the prognostic important indicators that must be taken into account when selecting stallions with the highest indicators of the quality of cryopreserved sperm.

Asinara male donkey (Equus africanus asinus var. Albina) and stallion (Equus ferus caballus) reproductive characteristics: Correlations between testicular blood supply and sperm production

The Asinara donkey is a rare small albino variety of feral donkey listed as "critical" by the Food and Agriculture Organization of the United Nations (FAO) and by the Domestic Animal Diversity Information System (DAD-IS) in 2022. The objectives of the present study were to evaluate the reproductive characteristics of healthy male Asinara donkeys through andrological evaluation and testicular echo-color-Doppler examination to assess correlations between testicular blood supply and sperm production and to compare it with horse reproductive characteristics. Eight healthy donkeys and stallions were subjected to semen collection and evaluation, ultrasound and Doppler evaluation of the testicles and serum testosterone determination. The testosterone concentrations in donkeys were 1.42 ± 0.69 ng/ml while in horses were 1.90 ± 0.63 ng/ml within the normal values reported in these species. Horses had greater mean and total testicular volume, semen gel-free volume, and total sperm number than donkeys (P < 0.001). Blood flow analysis of the testicular artery in both locations was a relatively simple procedure. The waveforms recorded in a convoluted location (-con) of the testicular artery were characteristic of the high-resistance vessel, while in the marginal location (-marg) were of low-resistance. It was also recorded a gradual decrease in peak systolic velocity (PSV), resistive index (RI), pulsatility index (PI), mean velocity (MV) and testicular arterial blood flow (TABF) values along the peripheral course of the testicular artery. Therefore, donkeys had greater PSV-con (P ≤ 0.01), PSV-marg (P < 0.01), MV-con (P < 0.001), MV-marg (P < 0.01), testicular arterial blood flow rate (TABFr-con) (P < 0.001) and TABFr-marg (P < 0.01) than horses. Conversely, pulsatility index (PI-con) was higher in horses (P < 0.05). The comparative analysis between the Doppler parameters and the reproductive/seminal characteristics showed a positive correlation of TTBF-con with Testosterone (ρ = 0.976, p < 0.01 and ρ = 0.905, p < 0.01 in donkeys and horses, respectively), with Total Testicular Volume (ρ = 0.952, p < 0.01 and ρ = 1.000, p < 0.01 in donkeys and horses, respectively), and with Total spermatozoa concentration (ρ = 0.905, p < 0.01 and ρ = 0.813, p < 0.05 in donkeys and horses, respectively). Additionally, only in the donkey there was a positive correlation with Semen gel free volume (ρ = 0.881, p < 0.01) and Spermatozoa concentration × ml (ρ = 0.786, p < 0.05). The testosterone concentration was positively correlated in both species with Total testicular volume (ρ = 0.976, p < 0.01 and ρ = 0.905, p < 0.01 in donkeys and horses, respectively) and Total sperm concentration (ρ = 0.881, p < 0.01 and ρ = 0.976, p < 0.01 in donkeys and horses, respectively). It was also correlated positively with Semen gel free volume (ρ = 0.857, p < 0.01) and spermatozoa concentration (ρ = 0.762, p < 0.05) in donkeys. Finally, the PI-con (ρ = − 0.786, p < 0.05) was negatively correlated with Semen gel free volume in the donkeys. The results of this study show that Total Testicular Blood Flow is the parameter, together with testosterone, most positively correlated with testicular volume and total sperm concentration in both species. Furthermore, differences in both reproductive and testicular hemodynamic characteristics between the Asinara donkey and stallion are highlighted, suggesting that caution should be taken in transferring knowledge from one species to another. New data on reproductive aspects, seminal characteristics, testicular blood flow perfusion in healthy Asinara donkeys were reported and these may represent reference value for further research or clinical applications.

Sperm parameters in the Great Dane: Influence of age on semen quality

Not all sires have sperm suitable for chilled or frozen storage, and success in artificial insemination (AI) varies highly among individual dogs and breeds. Fertilizing potential is further complicated as sperm quality declines with the aging process. Due to the rapidity of aging and senescence in large breed dogs, associated health and fertility changes may be observed over a shorter period, though this period remains undefined for any breed. Working with a population of purebred Great Danes (GD), our aims were (1) to characterize the distribution of a series of sperm parameters, (2) to distinguish sources of variation in sperm quality within this rapidly aging breed, and (3) to identify changes in sperm quality that may accompany aging. Ejaculates collected from young, middle-aged, and senior Great Dane dogs (n = 50) were evaluated for semen volume, total sperm number and viability, and reactive oxygen species (ROS), in addition to sperm morphology and kinematic parameters. Total testicular volume was also determined using ultrasonography. Testicular volume was not a predictor of sperm production in the GD, however, significant differences between coat colors were identified. Age was negatively associated with total motility, progressive motility, and amplitude of lateral head displacement (ALH) (p < .05). We identified significant relationships between GD male age and TM, PM, and immotility with −9.9%, −9.0%, and +8.3% change per year of age, respectively, which support the anecdotal reports of decline of the fertility with the advance of age in this breed. Sperm of younger GD dogs aged 12 ≤ x < 24 months had significantly higher TM, PM, ALH, and nonlinear motility (p < .05) than older dogs (x ≥ 48 months). High ROS levels were positively associated with TM and PM, average pathway distance (DAP) and straight line distance (DSL), average pathway velocity (VAP), straight line velocity (VSL), and the presence of hairpin tails (p < .05). While age and ROS have significant influences on sperm parameters in the GD, the influence of selection for breed specific phenotypes could help explain the functional significance of the diversity among GD males.

Varicocelectomy improves sperm parameters, sperm DNA integrity as well as the other critical semen features.

Varicocele is a major entity defined within male infertility. In this report we have studied the influence of laparoscopic varicocelectomy on semen quality, biochemical parameters of seminal plasma and sperm DNA fragmentation. In this study, the semen samples from patients with left-side varicocele of grade II-III before and after laparoscopic varicocelectomy were compared to healthy individuals and separated into three groups. The volume of semen, sperm concentration (106/ml), motility (%), viability (%) and normal morphology (%) were assessed. Total antioxidant capacity (TAC), catalase (CAT), superoxide dismutase (SOD) and malondialdehyde (MDA) together with other biochemical substances in seminal plasma as alpha-glucosidase (α-Glu), fructose (Fr) and citric acid (CA) were determined by ELISA method. The spermatozoa activity including ion-transports through sodium, potassium ATPase (Na+, K+-ATPase) and calcium, magnesium ATPase (Ca2+, Mg2+-ATPase) were determined by using spectrophotometry. In addition, flow cytometry method for detection of sperm DNA fragmentation was used. The results showed, that three months after varicocelectomy such intervention led to significant postoperative improvement in volume of semen (p<0.001), total sperm count (p<0.001), sperm motility (p<0.001) and spermatozoa with normal morphology (p<0.001). We found decreased α-Glu levels due to varicocelectomy (p<0.05). There has been shown a high positive correlation between Na+, K+-ATPase and Ca2+, Mg2+-ATPase activity with total number of spermatozoa (p<0.05). The TAC levels and DNA fragmentation values after varicocelectomy can be considered as significant indicators of good prognosis after surgical intervention. It has to be emphasized that α-Glu levels and total sperm count expressed statistically significant both positive and negative predictive values for semen assessment. Varicocelectomy may lead to significant improvement of semen quality although the observations must be correlated with clinical pregnancies observed thereafter.