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This study was aimed to determine the impact of different taurine and caffeine combinations on the motility, viability, and oxidative markers of chilled stallion spermatozoa. Each stallion semen sample was diluted in a ratio of 1:2, with various taurine and caffeine concentrations (2.5-7.5 mg/mL taurine + 0.625-1.25 mg/mL caffeine) dissolved in a conventional extender. The control samples (CON) were prepared by diluting ejaculate only using the conventional extender. The motility was analyzed using a CASA system at different time intervals (0, 6, 12, 24, and 30 h) and the viability was evaluated using a mitochondrial toxicity test (MTT) performed at the end of the incubation at 5 °C. The liquid part of experimental samples was separated by centrifugation after 30 h of incubation and underwent the evaluation of oxidative stress via the quantification of markers ferric reducing ability of plasma (FRAP) and total oxidant status (TOS). The samples that were treated with a combination of taurine and caffeine significantly improved the motility parameters, mainly after 12, 24, and 30 h of incubation. Samples extended with combination of taurine and caffeine neither compromise viability nor alterations of redox status. The results of this study describe the combination of taurine and caffeine as an optimal supplement for improving the quality of stallion semen during chilled storage.

This study aimed to characterize the bacterial profiles and their association with selected semen quality traits among two chicken breeds. Thirty Lohmann Brown and thirty ROSS 308 roosters were selected for semen quality estimation, including sperm motility, membrane and acrosome integrity, mitochondrial activity, and DNA fragmentation. The oxidative profile of the semen, including the production of reactive oxygen species (ROS), antioxidant capacity, protein, and lipid oxidation, were assessed as well. Moreover, the levels of pro-inflammatory cytokines, including tumor necrosis factor alpha (TNF-α), interleukins 1 and 6 (IL-1, IL-6) and C-reactive protein, as well as the concentrations of selected antibacterial proteins (cathelicidin, β-defensin and lysozyme) in the seminal plasma were evaluated with the enzyme-linked immunosorbent assay. The prevailing bacterial genera identified by the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were Citrobacter spp., Enterococcus spp., Escherichia spp. and Staphylococcus spp. While the bacterial load was significantly higher in the ROSS 308 line (p < 0.05), a higher number of potentially uropathogenic bacteria was found in the Lohmann Brown roosters. Antimicrobial susceptibility tests revealed a substantial resistance of randomly selected bacterial strains, particularly to ampicillin, tetracycline, chloramphenicol, and tobramycin. Furthermore, Lohmann Brown ejaculates containing an increased proportion of Escherichia coli presented with significantly (p < 0.05) elevated levels of TNF-α and IL-6, as well as ROS overproduction and lipid peroxidation. Inversely, significantly (p < 0.05) higher levels of β-defensin and lysozyme were found in the semen collected from the ROSS 308 roosters, which was characterized by a higher quality in comparison to the Lohmann Brown roosters. In conclusion, we emphasize the criticality of bacteriospermia in the poultry industry and highlight the need to include a more complex microbiological screening of semen samples designated for artificial insemination. [ABSTRACT FROM AUTHOR]

Artificial insemination, as an essential pillar of the modern poultry industry, primarily depends on the quality of semen collected from stud roosters. Since the collection and storage of ejaculates is not a sterile process, antimicrobial agents have become essential supplements to semen extenders. While the use of traditional antibiotics has been challenged because of rising bacterial resistance, natural biomolecules represent an appealing alternative because of their antibacterial and antioxidant properties. As such, this study strived to compare the effects of 50 μmol/L curcumin (CUR) with 31.2 µg/mL kanamycin (KAN) as a conventional antibiotic on rooster sperm quality in the presence of Salmonella enterica, Escherichia coli and Pseudomonas aeruginosa. Changes in sperm structural integrity and functional activity were monitored at 2 and 24 h of culture. Computer-assisted semen analysis revealed significant sperm motility preservation following treatment with KAN, particularly in the case of Salmonella enterica and Pseudomonas aeruginosa (p < 0.001) after 24 h. On the other hand, CUR was more effective in opposing ROS overproduction by all bacteria (p < 0.05), as determined by luminol-based luminometry, and maintained sperm mitochondrial activity (p < 0.001 in the case of Salmonella enterica; p < 0.05 with respect to Escherichia coli and Pseudomonas aeruginosa), as assessed by the fluorometric JC-1 assay. The TUNEL assay revealed that CUR readily preserved the DNA integrity of rooster sperm exposed to Salmonella enterica (p < 0.01) and Escherichia coli (p < 0.001). The bacteriological analysis showed higher efficiency of KAN in preventing the growth of all selected bacterial species (p < 0.0001) as opposed to CUR. In conclusion, CUR provided protection to rooster spermatozoa against alterations caused by uropathogens, most likely through its antioxidant activity. Hence, CUR supplementation to poultry semen extenders in combination with properly selected antibacterial substances may become an interesting strategy in the management of bacterial contamination during semen storage. [ABSTRACT FROM AUTHOR]

This study monitored the chemical and biochemical composition of bovine seminal plasma (SP). Freshly ejaculated semen (n = 20) was aliquoted into two parts. The first aliquot was immediately assessed to determine the sperm motion parameters. Another motility measurement was performed following an hour-long co-incubation of spermatozoa with SP at 6 °C. The other aliquot was processed to obtain the SP. Seminal plasma underwent the analyses of chemical composition and quantification of selected proteins, lipids and RedOx markers. Determined concentrations of observed parameters served as input data to correlation analyses where associations between micro and macro elements and RedOx markers were observed. Significant correlations of total oxidant status were found with the content of Cu and Mg. Further significant correlations of glutathione peroxidase were detected in relation to Fe and Hg. Furthermore, associations of chemical elements and RedOx markers and spermatozoa quality parameters were monitored. The most notable correlations indicate beneficial effects of seminal Fe on motility and Mg on velocity and viability of spermatozoa. On the contrary, negative correlations were registered between Zn and sperm velocity and seminal cholesterol content and motility. Our findings imply that seminal plasma has a prospective to be developed as the potential biomarker of bull reproductive health. [ABSTRACT FROM AUTHOR]

The purpose of present study was to observe the effect of resorcinol on bovine spermatozoa before and after cryopreservation. Fresh semen was obtained from six randomly chosen breeding bulls. Experimental samples were prepared by diluting the semen with six different concentrations of resorcinol (REZ1- 4; REZ2 - 2; REZ3 -1; REZ4 - 0.5; REZ5 - 0.25 and REZ6 -0.152 mg.ml-1). Experimental groups were compared against control group (REZK). Using CASA method spermatozoa motility was evaluated before cryopreservation and also after thawing, during incubation at 37°C in time periods 0, 5, 10, 15 and 60 minutes. Negative effect of in vitro resorcinol addition was observed in all analysed parameters (MOT, PRO, VCL, ALH and BCF) in groups REZ1, REZ2 and REZ3. In contrast, positive impact mainly on motility and progressive motility was found in experimental groups with the lowest resorcinol concentration addition (REZ4, REZ5 and REZ6) in comparison to the control group. After thawing significant effect of resorcinol on motility parameters were observed within each experimental group. Group with the highest concentration of resorcinol (REZ1) had markedly negative impact on evaluated motility parameters. Results of our study clearly confirm toxic effect of resorcinol on motility parameters of spermatozoa, which depend on concentration and time period. [ABSTRACT FROM AUTHOR]

Mistletoe (Viscum album L.) was previously described as an effective additive in the therapy of patients with cancer after medical therapy of the primary neoplasia. The defence activity and also direct cytotoxic stimulation was found to be upstanding for the anti-carcinogenic action. The main target of this study was to detect the effect of Viscum album pini L. at various concentration 1.0 (PA); 0.66 (PB); 0.33 (PC); 0.1 (PD); 0.066 (PE) and 0.033 (PF) mg/mL on the fine rabbit spermatozoa motility parameters at various time periods (0; 1; 2 and 3 h) during the cultivation in vitro. The CASA methodology was applied to estimate the spermatozoa motility parameters. The spermatozoa motility at the beginning of the experiment (Time 0) found decreased rate for examined doses of Viscum album pini compared to the control, but the statistical significant reduction was observed (p<0.01) only in the sample PD compared to groups PK (control). After 3 hours of in vitro cultivation interesting but not significant tendency of spermatozoa motility increase was found. Only in the group with the highest concentration (PA) a non-significant decrease was detected. Also, other parameters confirm this finding. After 3 hours of incubation, viability of rabbit spermatozoa showed decreased values in all doses of Viscum album pini in comparison to the control group. A significant decrease (p<0.01) of membrane integrity was found in group PA compared to control group. Also, in groups PB, PC, PD lower values in comparison to control group were detected. Acrosomal integrity showed very similar tendency in all groups. Our results detected any dose and/or time dependent significant effect of Viscum album pini on rabbit spermatozoa motility and viability in vitro. Positive impact on motility parameters after cultivation of rabbit semen with Viscum album pini was not found, but indications of changes on membrane integrity were observed. [ABSTRACT FROM AUTHOR]

Simple Summary: Overbreeding—and its associated increase in the chest areas of turkeys—has led to the loss of their natural ability to reproduce. Therefore, commercial production of turkey meat relies on artificial insemination. However, along with the physiology of the genital tract of turkeys, there is high potential for bacterial contamination of ejaculates. These bacteria may affect crucial semen quality parameters required for successful fertilization. As such, it is important to pay close attention to the bacteria present in turkey ejaculates and possible solutions to eliminate their adverse effects on avian spermatozoa. This study focused on the identification of naturally occurring bacteria in the reproductive fluid and impact on the quality of ejaculates obtained from the turkey breed British United Turkeys (BUT) Big 6 (n = 60). We determined possible relationships between the bacterial load and advanced sperm quality parameters that are important for effective artificial insemination and high fertility, as well as the concentration of selected antimicrobial proteins and pro-inflammatory markers of turkey semen. Sperm motility was assessed with computer-assisted sperm analysis (CASA), while the membrane and acrosome integrity were examined with smearing and staining methods. Reactive oxygen species (ROS) generation was quantified via luminometry, sperm DNA fragmentation was evaluated using the TUNEL assay, and the JC-1 assay was applied to evaluate the mitochondrial membrane potential. Cell lysates were prepared to investigate the extent of lipid and protein oxidation. Furthermore, levels of interleukins 1 and 6 (IL-1, IL-6), C-reactive protein, cathelicidin, and β-defensin were quantified in the seminal plasma using the ELISA method. The most dominant species identified by the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry was Escherichia coli, Proteus mirabilis, Staphylococcus lentus, and Citrobacter braakii. The bacterial load had a negative effect on the sperm motility (p < 0.001), as well as membrane (p < 0.05) and acrosome integrity (p < 0.01). A strong positive relationship between the bacterial load and DNA fragmentation (p < 0.001) was detected as well. Positive associations were recorded between the increasing presence of bacteria, ROS overgeneration (p < 0.001), and a subsequent oxidative damage to the proteins (p < 0.001) and lipids (p < 0.01). It was revealed that the antimicrobial peptides β-defensin (p < 0.001) and cathelicidin (p < 0.001) had a positive relationship with the motility. In contrast, pro-inflammatory markers, such as IL-1 (p < 0.001) and IL-6 (p < 0.001), had a negative impact on the motion behavior of turkey spermatozoa. Our results suggest that the semen quality may be notably affected by the bacterial quantity as well as quality. It seems that bacteriospermia is associated with inflammatory processes, oxidative stress, sperm structural deterioration, and a subsequent risk for a failed artificial insemination in turkey breeding. [ABSTRACT FROM AUTHOR]