Search Results (22)

The study of the relationship between testicular morphology and sperm quality is a pressing issue, for which molecular genetic approaches, including quantitative analysis of gene expression, are being implemented. The aim of this study was to identify correlations between the histomorphological structure of the testes, fresh sperm parameters, and the expression level of key spermatogenesis genes—TGFB2 and DMRT1—in roosters. The experiment was conducted on 10 Russian Snow White roosters aged 28–32 weeks. Sperm quality was assessed by volume, sperm concentration, total and progressive motility, and viability; histological analysis of the rooster testes was performed. The relative expression of the TGFB2 and DMRT1 genes in sperm was analyzed. Multiple correlation analysis of the data was conducted. A positive correlation was found between ejaculate volume and the number of spermatogonia (p = +0.651), a negative correlation between ejaculate volume and the number of second-order spermatocytes (p = −0.704), a negative correlation between the total cross-sectional area of the seminiferous tubules of the testes and sperm viability (p = −0.782), a negative correlation between the number of seminiferous tubules and the average diameter of their cross-section (p = −0.685), and a positive correlation between total and progressive sperm motility (p = +0.794). Analysis of TGFB2 and DMRT1 gene expression in sperm demonstrated a certain relationship between molecular genetic mechanisms and histomorphometric parameters. The expression level of the DMRT1 gene, which plays a key role in sex determination in birds during embryogenesis, had a number of negative correlations with such parameters as testicle weight (r = −0.782), total/progressive sperm motility (r = −0.552; r = −0.612), and viability (r = −0.552). Expression of the TGFB2 gene had no significant relationship with the studied parameters, but correlation analysis revealed a moderate positive relationship (r = +0.321) with DMRT1 gene expression. The data obtained indicate the expediency of integrating morphometric, cellular, and molecular analysis for an objective assessment of rooster reproductive function. Full article

Obesity is considered a metabolic disease, in which leptin is used as an indicator of energy in the body. This hormone, in turn, is related to the neuroendocrine regulation of the reproductive axis. However, leptin excess secretion due to obesity can have a negative effect on reproduction. Overweight and obesity were induced through high-calorie diets. Lee and gonadosomatic indices were determined to characterize the model and degree of reproductive development in the testis and epididymis. Sperm quality was analyzed using spermatobioscopy. Morphometry was analyzed through histological analysis. The changes described affect testicular function in hormone and sperm production. Exposure of 3-month-old male rats to diets with different fat contents (10% and 60%) induced both overweight and obesity. The animals showed morphological alterations, both testicular and epididymal, the latter being more sensitive to dietary changes by modifying the epididymal index, morphometric parameters (in both organs), and a decrease in cilia length. These changes induced a reduction in sperm viability, as well as an increase in malformed spermatozoa. In conclusion, both overweight and obesity have effects on male reproduction by modifying the morphology and physiology of reproductive organs. Full article

The study investigated the sperm motility and morphometry of pre-freeze and post-thaw boar epididymal semen cooled at increasing holding times at 18 °C. A total of 50 testes of heterogeneous boars were collected (5 testes/day) from the local abattoir and transported to the laboratory at 5 °C within 30 min after slaughter. Semen was retrieved from the caudal part of the epididymis using the slicing float-up method, diluted with Beltsville Thawing Solution extender, pooled in a 50 mL centrifuge tube/5 testes/day, and cooled at 18 °C. Following each holding time (0, 3, 6, 9, 12, 24, and 48 h), the cooled semen sample was re-suspended with Fraction A extender and stored at 5 °C for an additional 45 min. A cooled resuspended semen sample was then diluted with Fraction B extender, loaded into 0.25 mL straws, and frozen using liquid nitrogen vapour. Thawing was accomplished by immersing the semen straws in warm (37 °C) water for 1 min and the samples were evaluated for sperm motility and morphometry traits using the computer-assisted sperm analyzer system. The data were analyzed using variance analysis. Descriptive statistics were used to assess sperm morphometry, establishing the minimum and maximum values. Boar epididymal sperm survived for up to 48 h when held at 18 °C. Furthermore, the highest post-thawed sperm motility rates were observed in semen frozen after 3 h of holding time, with a sperm total motility of 85.9%, a progressive motility of 60.3%, and a rapid motility of 33.2%, as compared to other holding times (p < 0.05). The acceptable ranges for pre-freeze and post-thawed sperm morphology were head length (8.4–9.1 µm), width (4.4–4.8 µm), area (29.9–38.2 µm²), perimeter (20.1–23.7 µm), midpiece width (1.1–2.8 µm), and sperm shape, were consistent regardless of the holding time. A holding time of 3 h enhances the cryoresistance of sperm cooled at 18 °C. Therefore, these findings suggest that boar epididymal sperm can be effectively conserved and can maintain fertilization capability when cooled for 3 h at 18 °C before freezing. Full article

The present study aimed to evaluate an optimal method to transport and store drone sperm samples, as well as to characterize drone spermatozoa through sperm head morphometry. A total of 291 mature drones were used. We performed three experiments. In a first experiment, sperm variables were assessed under different incubation conditions (5 °C, 15 °C, and 37 °C with 5% CO₂). Results showed that sperm viability was optimally maintained at 15 °C (p ˂ 0.05). In the second experiment, the supplementation of extender with catalase (200 UI) improved (p ˂ 0.05) the sperm viability and motility during liquid storage at different incubation times. Finally, a morphometric analysis of sperm head was made: length 5.13 µm, width 0.85 µm, area 3.78 µm², perimeter 15.01 µm, acrosome length 3.50 µm. The variability in sperm head morphometry was calculated by coefficients of variation (CV) within- and between-drones. The CV within-drone was higher than the CV between-drones for all morphometric parameters regardless of hive origin, indicating a high degree of sperm pleomorphism. Full article

Studies revealed a global loss of genetic resources for local sheep breeds. Therefore, the current study aimed to introduce and highlight the progress made on Hungary’s existing gene conservation program (small Gene Bank). Furthermore, we evaluated breed (Tsigai, Cikta, and Racka), season, and individual variabilities (n = 24) of the pre-freeze and post-thaw semen stored in the Gene Bank to enhance the gene conservation of the breeds. The samples were cryopreserved manually, and post-thaw spermatozoa were analyzed for motility (CASA), viability, chromatin structure, and morphometry of the sperm nuclei. Ejaculate volume, spermatozoa concentration, subjective motility and standard motility, kinematic parameters, and spermatozoa’s head area standard deviation of the post-thaw samples differed significantly among breeds (p < 0.05). Season affected ejaculate volume, total spermatozoa number/ejaculate, STR, BCF, and ALH. We observed a significant (p < 0.001; 0.05) breed and season interaction on concentration, total spermatozoa number/ejaculate, VCL, LIN, WOB, spermatozoa’s head average perimeter and nucleus length (Tsigai and Cikta differed but were statistically the same as Racka). Similarly, season significantly (p < 0.05) affected the proportion of ejaculate suitable for freezing. There was a significant (p < 0.05) difference in kinematic parameters and viability among the rams across the breeds. The spermatozoa’s head morphometry of the Tsigai and Cikta breeds differed significantly (p < 0.05) among the rams. There were individual and breed differences in many spermatozoa quality parameters. The stored samples are of good quality, with more than 40% having intact membranes and low abnormal chromatin condensation. Full article

Our objectives were to investigate the effects of exogenous melatonin on testicular volume (TV), testicular blood flow (TBF), and semen quality in Bafra rams during the non-breeding season. One group of rams (MEL, n = 5) received a 36 mg melatonin implant twice, with 30 days in between, while the other group (CON, n = 5) served as the control. TBF, TV, and semen quality parameters were determined at three-week intervals starting three weeks before until twelve weeks after the first melatonin implant. Testicular blood flow was determined in the supratesticular (STA) and marginal testicular artery (MA) using color Doppler ultrasound. Semen was collected and evaluated, and the total oxidative status (TOS) and total antioxidative status (TAS) was determined using an ELISA. The MEL group had increased (p < 0.05) TV between the sixth and twelfth week after the start of treatment. Overall, the MEL group had lower resistance and pulsatility indexes (p < 0.05) between the third and ninth week, although there was no difference (p > 0.05) between the two groups in most semen quality parameters. However, TAS concentrations increased (p < 0.05) in the MEL group compared with the CON. The results of this study show that exogenous melatonin in the non-breeding season significantly increased both TBF and TV in Bafra rams. Therefore, giving rams implants with 36 mg melatonin twice at least one month prior to the non-breeding season is expected to improve testicular size and function and reproductive capacity. Full article

The objective of the present study was to determine the effects that the reproductive season has on the motility, kinematics, morphology, and sperm morphometry of Brahman bulls evaluated with a commercial CASA system. The experiment was carried out at the Costa Rica Institute of Technology from March to August 2021. A total of eight Brahman bulls were used. A total of 28 ejaculates were collected in the pre-mating period (PMP), during it (DMP), and after it (AMP) using an electroejaculator. The sperm concentration was measured with the Accuread photometer. The motility was measured using a Spermtrack^® counting chamber. The analyses were performed with the CASA-Mot ISAS^®v1 system. The morphology was analyzed using a microscope with a negative phase contrast objective. Morphometry was evaluated with the CASA-Morph. The sperm concentration did not present differences between the PMP and AMP; however, it was significantly higher than DMP (p > 0.05). Regarding the progressiveness variables, linearity on forward progression (LIN), straightness (STR), and wobble (WOB) were higher (p < 0.05) DMP. A kinematic principal component analysis grouped all the variables into three factors and an effect on the reproductive period was found (p < 0.05) in the parameters of the head and middle part of the sperm, such as width and perimeter, which were greater in the PMP. The length of the sperm head in the PMP and DMP did not show differences; however, both were larger (p < 0.05) than AMP. The insertion distance of the middle piece of the sperm was significantly greater than DMP. Finally, the PMP contained cells with a larger insertion angle (p < 0.05) than AMP. These findings are important to understand the implications of reproductive status on sperm quality and to consider them in andrological evaluations. Full article

Sperm morphology can predict the reproductive male fertilizing potential. This study aimed to determine the morphological and morphometric spermatozoa characteristics from guinea pigs subjected to different photoperiodic stimulation. Thirty F1 guinea pigs were randomly assigned to three photoperiodic treatments: FT1 (photoperiod with 10 Light/14 Dark LED light), FT2 (photoperiod with 10L/14D sunlight), and FT0 (room without direct light source). At 107 ± 9.8 days of age, sperm concentration and motility were higher in the FT0 and FT1 groups (p < 0.05); furthermore, there were no differences in nucleus length and ellipticity between the FT0 and FT1 groups, but the sperm of the FT1 group was higher in perimeter and nuclear area, while that of the FT0 group was higher in roughness, regularity, midpiece length, and tail (p < 0.01). Expanding acrosome (Type 2) was more frequent in the FT2 group, but there was variation in head measurements between all morphological categories. Pregnancy rate, calving age, and mating age were higher in the FT0 group; meanwhile, the FT1 group initiated successful matings earlier (p < 0.01). The FT0 group had a higher fertility rate, and the age of mating and first calving were earlier in the FT1 group than the FT0 group, but no pregnancies were reported for the FT2 group. Photoperiodic stimulation can increase the morphometric dimensions of guinea pig spermatozoa, favoring the reproductive characteristics, but sunlight could reduce their size due to heat stress. Full article

The objective was to characterize morphological, morphometric, and ultrastructural changes in rhea spermatozoa between the epididymis and the vas deferens. Sperm samples were collected from the reproductive tracts of seven adult individuals and evaluated for sperm characteristics using brightfield microscopy as well as ultrastructural features using scanning electron microscopy (SM). Mean sperm count tended to increase in the vas deferens (378.0 ± 135.0 × 10⁶) compared to the epididymis (201.0 ± 77.4 × 10⁶). Percentages of motile sperm grew from 37.0 ± 4.9% in the epididymis to 58.5 ± 7.7% in the vas deferens. The proportion of normal spermatozoa was 75.6 ± 1.8% and most common defects were bent tails (9.7 ± 0.9%). However, these proportions were not different between epididymis and vas deferens. SM analysis revealed further features of rhea spermatozoa. Normal rhea spermatozoa were threadlike with an acrosome (0.95 ± 0.0 µm), head (7.53 ± 0.01 µm), midpiece (2.08 ± 0.01 µm), and tail (30.7 ± 0.06 µm). Lengths of sperm acrosome, head, midpiece, and tail were longer in the vas deferens compared to the epididymis. Our findings suggest that rhea spermatozoa undergo a maturation process during the passage from the epididymis to the vas deferens. Full article

The alarming decline in sturgeon populations doubled by growing demands for sturgeon products, urge us to prevent the loss of these species. Fish stocking in natural habitats and developing fish farms are viable solutions, which can be successfully implemented with the help of reproductive biotechnologies. Despite the fact that semen cryopreservation is admittedly an important step for saving the Russian sturgeon, a reproducible standard method with good results has yet to be identified. Sperm quality assessment is essential for quantifying the impact of cryopreservation on spermatozoa. The purpose of our study was to provide additional information regarding semen cryopreservation and semen quality evaluation for the Russian sturgeon. Our study method is based on the use of two yolk-free extenders (with different cryoprotectants: DMSO and methanol) for freezing semen, using a simple freezing protocol. Parameters such as volume, concentration, motility, morphology and membrane integrity were evaluated. In conclusion, cryopreservation of Russian sturgeon spermatozoa using an extender containing methanol as cryoprotectant led to high egg fertilization percentages (72.67 ± 5.4%) even if the total motility values recorded for thawed semen were quite low (18–25%). Additionally, we identified two optimal stains for morphological studies and morphometry (Spermac stain kit and Trypan Blue Solution). Full article

The aim of the study was to assess the morphometry of sperm during storage of liquid boar semen at 17 °C. An attempt was also made to evaluate the suitability of three staining methods for assessment of boar sperm morphometry. The study was carried out on 20 Landrace boars. Semen was collected from the boars every 5 days by the manual method. Four ejaculates from each boar were analysed (80 ejaculates in total). Analyses were performed five times: at 1 h, 24 h, 48 h, 96 h, and 168 h after semen collection. Blisters with insemination doses were opened immediately before the analyses. From each insemination dose, smears were prepared for morphometric evaluation of sperm, which were stained by three methods (eosin-nigrosin—EN, eosin-gentian—EG, and SpermBlue—SB). Morphometric measurements of 15 randomly selected sperm with normal morphology were performed on each slide. The morphometric measurements included the following parameters: sperm head length, width, area, and perimeter; tail length; and total sperm length. The results of the morphometric measurements were used to calculate the head shape index. The morphometric dimensions of the sperm were shown to change during storage of semen at 17 °C. The extent of these changes, however, depended on the staining method used, as the three methods result in different morphometric dimensions of sperm, in the case of both the head and the tail. In the slides stained by the eosin-nigrosin method, the dimensions of the head and tail were smaller at every time of storage than in the slides stained by the SpermBlue and eosin-gentian methods. Full article

Background: Due to the increased use of titanium dioxide nanoparticles (TiO₂ NPs), the risks of their reprotoxic effect arise. This study anticipated examining the potential protective effects of GEO (geranium essential oil) components screened via GC/MS analysis against the reprotoxic impacts of TiO₂ NPs on male rats. Methods: Thirty-two adult male rats were randomly assigned to four groups: control, GEO (75 mg/kg bwt/orally/day/60 days), TiO₂ NPs (100 ppm/rat/IP/day/60 days), and TiO₂ NPs + GEO. After 60 days, hormonal assay, semen appraisal, lipid peroxidation, antioxidant enzymes, testis and prostate morphometry, and the steroidogenesis-related genes’ mRNA expressions were assessed. Results: The TEM and DLS results demonstrated that synthesized TiO₂ NPs are spherical with minimal aggregations polydispersed and varying in size from 50 to 100 nm. TiO₂ NPs IP injection-induced sperm abnormalities decreased the percent of motile sperms in the sperm count, reduced sex hormone levels, altered the testicular oxidant/antioxidant status and mRNA expression of steroid-related genes, and induced architectural alterations in testicular, epididymal, and prostate gland tissues. GEO significantly rescued the TiO₂ NPs-altered spermiogram, sex hormones, and antioxidant capacity, restored the tissue architectures, and enhanced steroidogenesis-related gene mRNA expression. Conclusions: These findings may significantly contribute to developing combinatorial treatments for infertility associated with various environmental and industrial xenobiotic exposures. Full article

The objective of the present study was to investigate whether fertility differences in bulls are reflected in variations of sperm quality when analysing only one ejaculate per male. Two experiments were performed. In the first experiment, frozen semen samples from 20 adult bulls were tested; 10 bulls had high field fertility and 10 bulls had low field fertility. Analyses of sperm motility, membrane integrity, and membrane–acrosome integrity with the ISAS3Fun method were performed. Sperm morphometry of the fluorescence sperm subpopulations obtained with the ISAS3Fun method was also analysed. Significant differences between high- and low-fertility groups were only found with the ISAS3Fun technique, specifically in sperm acrosome integrity, the proportion of spermatozoa with an intact acrosome and damaged membrane, and in sperm head width of spermatozoa with intact structures. Discriminant analyses allowed us to correctly classify 90% of sperm samples in their fertility group using sperm quality parameters. Given that only the results obtained with the ISAS3Fun technique were related to bull fertility, we performed a second experiment aimed to validate the efficacy of this technique to detect the acrosomal integrity of bull spermatozoa, comparing them with the conventional FITC-PNA/propidium iodide (PNA/PI) combination under capacitating conditions. The results indicated that the ISAS3Fun combination provided an accurate assessment of both viability and acrosomal integrity for ejaculated spermatozoa, while the PNA/PI combination underestimated the extension of acrosomal damage due to false negatives. It was concluded that the simultaneous assessment of sperm plasma membranes and acrosome integrity with the ISAS3Fun method is precise and seems to have a greater potential to discriminate between high- and low-fertility bulls than more conventional in vitro sperm quality tests. Full article

The adverse effects of maternal pesticides exposure on the progeny is very well established. However, the impact of paternal exposure to pesticides such as Fenitrothion (FNT) on the histomorphometry of progeny’s organs in unexposed mothers are much less well studied. Therefore, this study aims to evaluate the effects of paternal FNT exposure on the sperm quality of the parent rat and its effects on the histomorphometry of the progeny’s organs. Randomly, male Sprague Dawley rats (n = 24) categorized as F0 were distributed equally into three groups namely Control, FNT-10, and FNT-20. Control received 1 mL/kg corn oil while FNT-10 and FNT-20 received 10 mg/kg and 20 mg/kg of FNT, respectively, via oral force feeding for 28 consecutive days. At the end of the study, male rats were mated with unexposed female rats and the male rats were sacrificed to obtain sperm for sperm characterization and DNA damage evaluation. Meanwhile, the rats’ progeny (F1) namely pControl, pFNT-10, and pFNT-20 were left to grow until postnatal day 70 before being sacrificed to obtain the matured organs for histology and morphometric analysis. Our results showed that both doses of FNT reduced sperm quality and caused DNA fragmentation in F0 rats compared with the control group (p < 0.05). The number of Leydig cells as well as the diameter of the seminiferous tubules and glomerulus of the pFNT-20 group had significantly decreased (p < 0.05) compared with the pControl group. The Bowman’s space of the pFNT-20 group had significantly increased (p < 0.05) compared with the pFNT-10 and pControl groups. Therefore, paternal exposure to FNT reduced the sperm quality and increased sperm DNA fragmentation in F0 male Sprague Dawley rats and altered the histology and morphometry of the selected organs in the F1 progeny. Full article

Bottlenose dolphin (Tursiops truncatus) males follow many reproductive strategies to ensure their paternity. However, little is known about the sperm traits, including morphometric features, that contribute to their reproductive success. Our aim was to study dolphin sperm morphometry (a total of 13 parameters) in two adult males to evaluate (i) presumptive sperm subpopulations, (ii) the correlation of sperm morphometry with testosterone levels and (iii) the effect of refrigerated storage on the sperm morphometry. Sperm populations were classified into four principal components (PCs) based on morphometry (>94% of cumulative variance). The PCs clustered into two different sperm subpopulations, which differed between males. Furthermore, the levels of serum testosterone were positively correlated with the length of the midpiece but negatively correlated with head width and the principal piece, flagellum and total sperm lengths. Most of the sperm morphometric parameters changed during the storage period (day 1 vs. day 7), but only the principal piece length was affected by the storage temperature (5 °C vs. 15 °C). This is the first study to identify dolphin sperm subpopulations based on morphometry and the influence of serum testosterone and refrigeration on sperm morphometry. Full article