The aim of this study was to determine the effect of semen extender supplementation with 25 or 50 μM of zinc nanoparticles (ZnNPs) or manganese nanoparticles (MnNPs) on turkey spermatozoa preserved in a liquid state. Twenty turkey ejaculates were obtained from twenty healthy males. The collected semen was preserved at 4 °C for 48 h with or without NPs. Selected qualitative and quantitative parameters of sperm (motility, plasma membrane activity, mitochondrial membrane potential (MMP) and the percentage of sperm demonstrating NO and SOD activity) were examined after 2, 24 and 48 h of storage. Sperm motility and MMP decreased in semen preserved with ZnNPs at each time point of the analysis. However, all spermatozoa remained viable throughout storage. In contrast, membrane integrity and mitochondria activity (p ≤ 0.05) increased, and the highest SOD activity (p ≤ 0.05) was observed in semen preserved with MnNPs. The addition of MnNPs to the semen extender could potentially improve the parameters of turkey semen during prolonged storage.

Metalloproteinases (MMP)s regulate developmental processes, control angiogenesis and wound healing, participate in the formation of immune receptors, and are expressed in stem cells. Retinoic acid (RA) is a potential modulator of these proteinases. The aim was to determine (1) MMPs’ action in antler stem cells (ASCs) before and after differentiation into adipo-, osteo-, and chondrocytes and (2) the effect of RA on modifying MMP action in ASCs. Antler tissue from pedicle was collected approximately 40 days after antler casting, post mortem from healthy breeding five year old males (N = 7). The cells were isolated from the pedicle layer of periosteum after skin separation and cultured. The pluripotency of the ASCs was evaluated by mRNA expression for NANOG, SOX2, and OCT4. ASCs were stimulated with RA (100nM) and differentiated for 14 days. The MMP (1–3) and TIMP(1–3) (tissue inhibitor of MMPs) mRNA expression was determined in the ASCs, their concentrations in the ASCs and the medium after RA stimulation as well as profiles of mRNA expression for MMPs: 1–3 and TIMPs: 1–3 during differentiation of ASC to osteocytes, adipocytes and chondrocytes. RA increased MMP-3 and TIMP-3 mRNA expression and output (P < 0.05) and not influenced on MMP-1 and TIMP-1 mRNA expression and output in ASC (P > 0.05). Depending on differentiation of ASC to osteocytes, adipocytes or chondrocytes, MMPs`and TIMPs`expression profile fluctuates for all studied proteases and its inhibitors. The studies demand continuation considering the role of proteases in stem cells physiology and differentiation. The results may be relevant for the study of cellular processes during the cancerogenesis of tumor stem cells.

Seminal plasma is rich in proteins originating from various male reproductive organs. The phosphorylation of these proteins can significantly impact sperm motility, capacitation, and acrosome reaction. Phosphoproteomics identifies, catalogues, and characterizes phosphorylated proteins. The phosphoproteomic profiling of seminal plasma offers valuable insights into the molecular mechanisms that influence semen quality and male fertility. Thus, the aim of this study was a phosphoproteomic analysis of white and yellow turkey seminal plasma. The experimental material consisted of 100 ejaculates from BIG-6 turkeys between 39 and 42 weeks of age. The collected white and yellow turkey seminal plasmas were analyzed for total protein content; the activity of selected enzymes, i.e., alkaline phosphatase (ALP), acid phosphatase (ACP), superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase (CAT); and the content of reduced glutathione (GSH) and malondialdehyde (MDA). Phosphoproteins were isolated from white and yellow seminal fluids, and the resulting protein fractions were separated by SDS-PAGE and Western blotting. Phosphorylated residues were immunodetected, and the isolated phosphoproteins were identified (nano LC-MS/MS). Yellow seminal plasmas were characterized by higher levels of total protein, GSH, and MDA, as well as higher levels of ALP, ACP, and GPx activity. There were no significant differences in the activity of SOD and CAT. A total of 113 phosphoproteins were identified in turkey seminal fluids. The functional analysis demonstrated that these phosphoproteins were mainly involved in oocyte fertilization, organization and metabolism of the actin cytoskeleton, amplification of the intracellular signal transduction pathway, general regulation of transport, vesicular transport, proteome composition of individual cellular compartments, and the organization and localization of selected cellular components and macromolecules. Increased phosphorylation of the fractions containing proteins encoded by SPARC, PPIB, TRFE, QSOX1, PRDX1, PRDX6, and FASN genes in white plasmas and the proteins encoded by CKB, ORM2, APOA1, SSC5D, RAP1B, CDC42, FTH, and TTH genes in yellow plasmas was observed based on differences in the optical density of selected bands. The obtained results indicate that the phosphorylation profiles of turkey seminal plasma proteins vary depending on the type of ejaculate.

Yellow semen syndrome (YSS) is an increasingly common reproductive health problem in male turkeys. This condition is characterised by a yellow discolouration of semen, often linked to decreased semen quality and fertility. Yellow semen syndrome poses a significant concern due to its negative impact on the reproductive performance of turkeys. Phosphorylation is one of the major post-translational modifications of proteins. A better understanding of the function of the sperm phosphoproteome is crucial for the advancement of reproductive biology and the development of therapies for male infertility. Spermatozoa from semen samples with YSS were characterised by lower levels of malondialdehyde (MDA), reduced plasma membrane integrity (PMI), and decreased mitochondrial membrane potential (MMP). However, these samples showed increased antioxidant enzyme activity and an elevated glutathione (GSH) content. Yellow sperm also had a lower percentage of viable cells and a higher proportion of apoptotic and necrotic cells. The phosphoproteins identified in turkey sperm play key roles in sperm maturation, the development of a functional motility apparatus, efficient cellular metabolism, protection against oxidative stress, and successful fertilisation of an egg. Yellow semen syndrome altered the phosphorylation of turkey sperm proteins on serine, threonine (p ≤ 0.05), and tyrosine residues, which could have influenced the metabolism and physiology of spermatozoa in yellow semen samples, thus affecting their reproductive potential. These findings highlight the impact of YSS on sperm function, including phosphorylation-dependent processes that are crucial for reproduction.

The study aimed to examine the parameters of turkey ejaculates (n=40) of high and low quality (HQ and LQ, respectively), preserved with the addition of taurine (TAU; 5 mM, 10 mM) and ergothioneine (EGT; 5 mM, 10 mM) for 48 h at 5°C in a liquid state. The motility, plasma membrane integrity (PMI), mitochondrial function, apoptotic and necrotic spermatozoa percentage, and sperm cells that generated NO were determined after 2, 24, and 48 h of storage. The preserved semen was also analysed for the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT), glutathione (GSH) content, and malondialdehyde (MDA) production. Taurine, in both doses, may improve the antioxidant status of stored turkey semen as well as sperm motility, viability and functionality regardless of ejaculate quality, as manifested by increased SOD and CAT activities, reduced MDA levels, and enhanced sperm characteristics, i.e. plasma membrane integrity (PMI), mitochondrial membrane potential (MMP), total motility (TMOT), and progressive motility (PMOT). In turn, the addition of EGT increased GSH content in the external environment and suppressed lipid peroxidation in turkey spermatozoa, in particular those from low-quality (LQ) ejaculates. However, it appears that EGT could deliver more satisfactory results when added at lower concentrations.

The aim of the study was to assess the impact of solarium light therapy on selected biological and biochemical parameters of peripheral blood in recreational horses. The study involved 10 horses divided into two groups of young (aged 5 to 7 years) and old (aged 14 to 19 years) individuals. All animals participated in light therapy sessions every other day. Blood was sampled three times during the study: before the treatment, after five light sessions, and after ten light sessions. Morphological parameters, the activity of antioxidant enzymes, TAS values, and the levels of glutathione (GSH), vitamin D₃, vitamin C, and malondialdehyde (MDA) were measured in the whole blood. Light therapy contributed to an increase in MCV, HDW, MCVr, CHr and MPV indices, and simultaneously a decrease in the basophil counts, MCHC, RDW and CHCMr indices in both groups of horses (p ≤ 0.05). At the same time reticulocytes fell in older whereas white blood cells and monocytes counts expanded in younger individuals. The treatment also increased the activity of glutathione reductase (GR) and glutathione peroxidase (GPx) in young but decreased the activity of mentioned enzymes in blood plasma of old horses. The total antioxidant status (TAS) of the blood plasma rose progressively, whereas GSH levels declined in all individuals. Moreover, vitamin D₃ levels did not change, whereas vitamin C levels gradually decreased during the experiment. The therapy also helped to reduce levels of MDA in the blood plasma, especially of older horses (p ≤ 0.05). In turn, GPx and GR activities as well as MDA levels significantly declined, whereas GSH levels notably elevated in erythrocytes (p ≤ 0.05). Solarium light therapy appears to have a beneficial impact on the morphological parameters and antioxidant status of blood in recreational horses in the winter season. However, the observed results could in part be attributed to the natural physiological adaptation of each individual organism to the treatment.