Uncaria tomentosa (UT) or cat's claw, is a vine belonging to the Rubiaceae family and native to South and Central America. Various parts of the plant, including bark, showed many therapeutic activities (e.g., antioxidant and antibacterial), but the in vitro effects on gametes have still not been investigated. During boar semen storage for artificial insemination purposes, oxidative stress and bacterial contamination negatively affect sperm quality. In this study, we evaluated the tolerance of boar sperm to UT ethanolic extract at four concentrations (1.6 to 0.025 μg/mL). The analyses were carried out on sperm samples under oxidative stress, induced by H2O2 and Fe2+/Ascorbate, and during 96 h of semen storage at 17°C. The antibacterial activity of the extract (1,024 to 8 μg/mL) was tested against commercial strains and bacteria isolated from the semen. The treatments ranging from 0.4 to 0.025 μg/mL protected sperm membrane (p < 0.05) and preserved some kinetic parameters in samples under oxidative stress (Fe2+/Ascorbate). During semen storage, the extract did not show any cytotoxicity, and mean values of some sperm parameters were higher than the control group, although not significant (p > 0.05). All tested Gram-positive bacteria exhibited growth inhibition. The most frequently isolated Gram-negative bacteria from semen (i.e., Citrobacter koseri, Pseudomonas aeruginosa, Stenotrophomonas maltophilia) also showed complete growth inhibition, while the remaining strains showed a partial decrease in growth. Taken together, our findings show that Uncaria tomentosa is a promising plant-based additive for boar semen storage.

• Keywords
• antibiotics, antioxidant, cat's claw, oxidative stress, pig, plant extract, secondary metabolites, semen storage,
• Publication type
• Journal Article MeSH

Hydrogen sulfide (H2S) is an endogenously produced signaling molecule that belongs to the group of gasotransmitters along with nitric oxide (NO) and carbon monoxide (CO). H2S plays a pivotal role in male reproductive processes. It is produced in various tissues and cells of the male reproductive system, including testicular tissue, Leydig and Sertoli cells, epididymis, seminal plasma, prostate, penile tissues, and sperm cells. This review aims to summarize the knowledge about the presence and effects of H2S in male reproductive tissues and outline possible therapeutic strategies in pathological conditions related to male fertility, e. g. spermatogenetic disorders and erectile dysfunction (ED). For instance, H2S supports spermatogenesis by maintaining the integrity of the blood-testicular barrier (BTB), stimulating testosterone production, and providing cytoprotective effects. In spermatozoa, H2S modulates sperm motility, promotes sperm maturation, capacitation, and acrosome reaction, and has significant cytoprotective effects. Given its vasorelaxant effects, it supports the erection of penile tissue. These findings suggest the importance and therapeutic potential of H2S in male reproduction, paving the way for further research and potential clinical applications.

• Keywords
• antioxidant, erectile function, hydrogen sulfide, male reproduction, sperm, testes, vasorelaxation,
• MeSH
• Erectile Dysfunction drug therapy   metabolism   MeSH
• Humans MeSH
• Infertility, Male metabolism   drug therapy   MeSH
• Genitalia, Male metabolism   drug effects   MeSH
• Reproduction * drug effects   physiology   MeSH
• Spermatogenesis * drug effects   MeSH
• Spermatozoa drug effects   metabolism   MeSH
• Hydrogen Sulfide * metabolism   pharmacology   MeSH
• Testis metabolism   drug effects   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Review MeSH
• Names of Substances
• Hydrogen Sulfide * MeSH

BACKGROUND: Antimicrobial resistance (AMR) is nowadays a major emerging challenge for public health worldwide. The over- and misuse of antibiotics, including those for cell culture, are promoting AMR while also encouraging the research and employment of alternative drugs. The addition of antibiotics to the cell media is strongly recommended in sperm preservation, being gentamicin the most used for boar semen. Because of its continued use, several bacterial strains present in boar semen have developed resistance to this antibiotic. Antimicrobial peptides and proteins (AMPPs) are promising candidates as alternative antibiotics because their mechanism of action is less likely to promote AMR. In the present study, we tested two AMPPs (lysozyme and nisin; 50 and 500 µg/mL) as possible substitutes of gentamicin for boar semen preservation up to 48 h of storage. RESULTS: We found that both AMPPs improved sperm plasma membrane and acrosome integrity during semen storage. The highest concentration tested for lysozyme also kept the remaining sperm parameters unaltered, at 48 h of semen storage, and reduced the bacterial load at comparable levels of the samples supplemented with gentamicin (p > 0.05). On the other hand, while nisin (500 µg/mL) reduced the total Enterobacteriaceae counts, it also decreased the rapid and progressive sperm population and the seminal oxidation-reduction potential (p < 0.05). CONCLUSIONS: The protective effect of lysozyme on sperm function together with its antimicrobial activity and inborn presence in body fluids, including semen and cervical mucus, makes this enzyme a promising antimicrobial agent for boar semen preservation.

• Keywords
• Antimicrobial resistance, Boar semen, Gentamicin, Lysozyme, Nisin, Sperm function,
• MeSH
• Acrosome drug effects   MeSH
• Anti-Bacterial Agents * pharmacology   MeSH
• Antimicrobial Peptides pharmacology   MeSH
• Cell Membrane drug effects   MeSH
• Gentamicins pharmacology   MeSH
• Muramidase * pharmacology   MeSH
• Nisin * pharmacology   MeSH
• Swine MeSH
• Semen drug effects   MeSH
• Spermatozoa drug effects   MeSH
• Semen Preservation * veterinary   methods   MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Anti-Bacterial Agents * MeSH
• Antimicrobial Peptides MeSH
• Gentamicins MeSH
• Muramidase * MeSH
• Nisin * MeSH

BACKGROUND: Hydrogen sulfide (H2S) donors are crucial tools not only for understanding the role of H2S in cellular function but also as promising therapeutic agents for oxidative stress-related diseases. This study aimed to explore the effect of amino acid-derived N-thiocarboxyanhydrides (NTAs), which release physiological H2S levels in the presence of carbonic anhydrase, on porcine sperm function during short-term incubation with and without induced oxidative stress. For this purpose, we employed two H2S-releasing NTAs with release half-lives (t1/2) in the range of hours that derived from the amino acids glycine (Gly-NTA) or leucine (Leu-NTA). Because carbonic anhydrase is crucial for H2S release from NTAs, we first measured the activity of this enzyme in the porcine ejaculate. Then, we tested the effect of Gly- and Leu-NTAs at 10 and 1 nM on sperm mitochondrial activity, plasma membrane integrity, acrosomal status, motility, motile subpopulations, and redox balance during short-term incubation at 38 °C with and without a reactive oxygen species (ROS)-generating system. RESULTS: Our results show that carbonic anhydrase is found both in spermatozoa and seminal plasma, with activity notably higher in the latter. Both Gly- and Leu-NTAs did not exert any noxious effects, but they enhanced sperm mitochondrial activity in the presence and absence of oxidative stress. Moreover, NTAs (except for Leu-NTA 10 nM) tended to preserve the sperm redox balance against the injuries provoked by oxidative stress, which provide further support to the antioxidant effect of H2S on sperm function. Both compounds also increased progressive motility over short-term incubation, which may translate into prolonged sperm survival. CONCLUSIONS: The presence of carbonic anhydrase activity in mammalian spermatozoa makes NTAs promising molecules to investigate the role of H2S in sperm biology. For the first time, beneficial effects of NTAs on mitochondrial activity have been found in mammalian cells in the presence and absence of oxidative stress. NTAs are interesting compounds to investigate the role of H2S in sperm mitochondria-dependent events and to develop H2S-related therapeutic protocols against oxidative stress in assisted reproductive technologies.

• Keywords
• Bioenergetics, Gasotransmitters, H2S-releasing agents, Reproductive biotechnologies, Sperm lifespan,
• MeSH
• Amino Acids * metabolism   MeSH
• Carbonic Anhydrases * metabolism   pharmacology   MeSH
• Mitochondria MeSH
• Oxidative Stress MeSH
• Swine MeSH
• Reactive Oxygen Species metabolism   MeSH
• Mammals MeSH
• Seeds metabolism   MeSH
• Spermatozoa MeSH
• Animals MeSH
• Check Tag
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Amino Acids * MeSH
• Carbonic Anhydrases * MeSH
• Reactive Oxygen Species MeSH

Persulfidation contributes to a group of redox post-translational modifications (PTMs), which arise exclusively on the sulfhydryl group of cysteine as a result of hydrogen sulfide (H2S) action. Redox-active molecules, including H2S, contribute to sperm development; therefore, redox PTMs represent an extremely important signalling pathway in sperm life. In this path, persulfidation prevents protein damage caused by irreversible cysteine hyperoxidation and thus maintains this signalling pathway. In our study, we detected both H2S and its production by all H2S-releasing enzymes (cystathionine γ-lyase (CTH), cystathionine β-synthase (CBS), and 3-mercaptopyruvate sulfurtransferase (MPST)) in male reproduction, including spermatozoa. We provided evidence that sperm H2S leads to persulfidation of proteins, such as glyceraldehyde-3-phosphate dehydrogenase, tubulin, and anchor protein A-kinase. Overall, this study suggests that persulfidation, as a part of the redox signalling pathway, is tightly regulated by enzymatic H2S production and is required for sperm viability.

• MeSH
• Cystathionine gamma-Lyase metabolism   MeSH
• Cysteine metabolism   MeSH
• Humans MeSH
• Reproduction MeSH
• Semen metabolism   MeSH
• Hydrogen Sulfide * metabolism   MeSH
• Check Tag
• Humans MeSH
• Male MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Names of Substances
• Cystathionine gamma-Lyase MeSH
• Cysteine MeSH
• Hydrogen Sulfide * MeSH

Various recent studies dedicated to the role of nitric oxide (NO) and hydrogen sulfide (H2S) in somatic cells provide evidence for an interaction of the two gasotransmitters. In the case of male gametes, only the action of a single donor of each gasotransmitter has been investigated up until today. It has been demonstrated that, at low concentrations, both gasotransmitters alone exert a positive effect on sperm quality parameters. Moreover, the activity of gaseous cellular messengers may be affected by the presence of oxidative stress, an underlying condition of several male reproductive disorders. In this study, we explored the effect of the combination of two donors SNP and NaHS (NO and H2S donors, respectively) on boar spermatozoa under oxidative stress. We applied NaHS, SNP, and their combination (DD) at 100 nM concentration in boar spermatozoa samples treated with Fe2+/ascorbate system. After 90 min of incubation at 38 °C, we have observed that progressive motility (PMot) and plasma membrane integrity (PMI) were improved (p < 0.05) in DD treatment compared to the Ctr sample under oxidative stress (CtrOX). Moreover, the PMot of DD treatment was higher (p < 0.05) than that of NaHS. Similar to NaHS, SNP treatment did not overcome the PMot and PMI of CtrOX. In conclusion, for the first time, we provide evidence that the combination of SNP and NaHS surmounts the effect of single-donor application in terms of PMot and PMI in porcine spermatozoa under oxidative stress.

• Keywords
• boar spermatozoa, gasotransmitter interaction, hydrogen sulfide, nitric oxide, oxidative stress,
• Publication type
• Journal Article MeSH

Oxidative stress occurs when the levels of reactive oxygen species (ROS) overcome the antioxidant defenses of the organism, jeopardizing several biological functions, including reproduction. In the male reproductive system, oxidative stress not only impairs sperm fertility but also compromises offspring health and survival, inducing oxidative damage to lipids, proteins and nucleic acids. Although a clear link between oxidative stress and male fertility disorders has been demonstrated in humans and laboratory rodents, little information is available about the implications of impaired redox homeostasis in the male fertility of domestic and wild animals. Therefore, this review aims to provide an update regarding the intrinsic and extrinsic factors that are associated with oxidative stress in the male reproductive system and their impact on the reproductive performance of domestic and wild animals. The most recent strategies for palliating the detrimental effects of oxidative stress on male fertility are reviewed together with their potential economic and ecological implications in the livestock industry and biodiversity conservation.

• Keywords
• ROS, antioxidant, infertility, livestock, semen, sperm dysfunction, wildlife,
• Publication type
• Journal Article MeSH
• Review MeSH

Tomcats are considered to be adults at 1 year of age, although many reach sexual maturity at an earlier age. Nevertheless, we still know little about whether the spermatogenic activity and sperm quality of mature under one-year-old tomcats differ from those of tomcats that are over one-year-old. This study aims to evaluate the spermatogenic activity, sperm traits, and their relationships in mature tomcats at two different ages. Sixteen tomcats showing complete spermatogenesis and spermatozoa in their epididymal caudae were used and classified according to their age as post-pubertal (<1 year old) or adult (˃1 year old). Our results show that adult cats have higher epididymal sperm concentration and lower coefficient of variation in sperm head width and ellipticity than post-pubertal cats. However, they do not differ in their testicular and epididymal mass, spermatogenesis, and sperm traits such as motility, mitochondrial activity, morphology, morphometry, as well as plasma membrane, acrosome, and DNA integrity. Reduced intra-male variation of sperm head ellipticity is associated with higher testis mass, epididymis mass, and sperm concentration. Interestingly, low intra-male variation in sperm head size is associated with increased Sertoli cell function and reduced post-meiotic germ cell loss. These findings increase our knowledge about feline reproductive physiology and provide new insights into the functional significance of low intra-male variation in sperm size and shape in tomcats.

• Keywords
• feline, intra-male variation, sexual maturity, sperm morphometry, spermatogenesis, spermatozoa, testis,
• MeSH
• Biological Variation, Population * MeSH
• Epididymis growth & development   MeSH
• Fertility MeSH
• Sperm Head physiology   MeSH
• Cats MeSH
• Sperm Motility MeSH
• Sperm Count MeSH
• Sexual Development MeSH
• Spermatogenesis * MeSH
• Testis growth & development   MeSH
• Age Factors MeSH
• Cell Size * MeSH
• Animals MeSH
• Check Tag
• Cats MeSH
• Male MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH