Evaluation of the effectiveness of the use of exosomes in the regulation of the mitochondrial membrane potential of frozen/thawed spermatozoa.

Numerous studies confirm the involvement of extracellular vesicles (EVs) in the regulation of physiological processes of mammalian sperm cells. It has been proven that they take part in the processes of capacitation, acrosonmal reaction, and anti-oxidation. Despite growing interest in the biomedical potential (including the search for new reproductive biomarkers) of EVs, the role of extracellular seminal vesicles in maintaining semen quality during cryopreservation has not yet been established. Therefore, the objective of this experiment was to evaluate the effectiveness of the use in the regulation of the mitochondrial membrane potential of bovine sperm and to explain the mechanisms of EV action during cell cryopreservation. Exosomes were isolated from bull semen plasma, measured, and used for extender supplementation. Semen samples were collected from Simmental bulls, diluted, and pre-evaluated. Then they were divided into equal fractions that did not contain EVs or were supplemented with 0.75; 1.5 and 2.25 mg/ml of EVs. The test samples were frozen/thawed and the mitochondrial membrane potential, DNA integrity, and viability were evaluated. EVs have been established to have a positive effect on cryopreserved sperm structures. The most favourable level of EVs was 1.5 mg / ml, which can be successfully to improve cell cryostability during freezing/thawing. In this study, exosomes isolated from the sperm plasma and supplemented with a concentrated dose in the extender for sperm freezing were shown to significantly improve cryostability of cells by supporting the potentials of the mitochondrial membrane and protecting the cytoplasmic membrane of spermatozoa.

Relationship between Milk Protein Polymorphism and Selected Cows' Reproductive Indices.

This research sought to determine the effect of selected milk protein polymorphisms on the reproduction parameters of 598 black and white Polish Holstein-Friesian cattle. The analyzed genes were kappa-casein (CSN3) and beta-lactoglobulin (BLG). The following reproductive indexes were selected: the age at first calving, the interpregnancy period, the calving interval, and the insemination index. The influence of the identified genotypes on the course of parturition in cows was also analyzed. Source data were collected for each cow from the beginning of the herd life and reproduction to its culling from the herd or the end of its third lactation. Data on the age at first calving, the amount of semen portions for artificial insemination (insemination index), the interpregnancy period, and the calving interval for each cow were also collected. A contingency analysis was performed through contingency tables using a Pearson's chi-squared test for each CSN3 and BLG genotype. The results show that the BB genotype of the kappa-casein gene was associated with the most favorable values of reproductive indicators. In the case of the calving interval, the values were significantly more favorable than those of other genotypes (p ≤ 0.05). No effect of beta-lactoglobulin polymorphism on the analyzed reproductive indices was observed. On the other hand, in the case of the interpregnancy period, significant statistical differences were obtained between the AA and BB genotypes. The analyzed kappa-casein and beta-lactoglobulin genotypes did not significantly influence the course of parturition in cows. To conclude, the genotype polymorphism BB CSN3 is the most favorable for the performance of the cows in the examined herd.

Examination of the haematological profile of pregnant Polish Holstein-Friesian black-and-white cattle in the early stage.

Introduction: Cattle health and welfare are monitored via the analysis of the haematological profile, and it shows cattle's ability to adapt to changing environmental conditions, pregnancy and lactation; profile changes also indicate reproductive disorders. The literature lacks reports of the examination of the haematological profile in cows up to the 50th day of pregnancy (dop). Therefore, this research examined that in cows up to this pregnancy stage. Material and Methods: A total of 101 Polish Holstein-Friesian black-and-white cows were divided into groups. The control groups consisted of non-pregnant heifers (group C00) and non-pregnant cows (group C0), and the experimental groups were pregnant heifers (group T1 at dop ≤ 28 and group T2 at dop ≥ 29-dop < 45) and pregnant cows (group T3 at dop ≥ 29-dop ≤ 50). In addition, the T3 group was divided into cows pregnant for up 45 dop and cows between 45 and 50 dop. Blood samples were collected in March and April 2021 from each animal and analysed. A transrectal ultrasound examination was performed to detect and confirm pregnancy. Results: Statistically significant differences (P ≤ 0.01) between the group of cows at dop < 45 dop and those at dop ≥ 45-dop ≤ 50 dop were noted in granulocyte percentage (GRA%), white and red blood cell counts (WBC/RBC), platelets (PLT), platelet distribution width (PDW), haematocrit (HCT) and lymphocyte percentage (LYM%). No statistically significant differences were found in the mean corpuscular haemoglobin, monocytes (MON), monocyte percentage (MON%), mean platelet volume (MPV), thrombocrit or red blood cell distribution width (RDW). Similar statistically significant differences (P ≤ 0.01) emerged between the groups of heifers in PLT, GRA, RBC, lymphocytes, LYM% and HCT, and no significant differences were found between MPV, MON, MON% or RDW. Conclusion: Examining the haematological profile in high-yielding cattle is vital in maintaining herd profitability and high reproduction, which depend on the quick diagnosis of disorders facilitated by haematology. This study analysed the haematology profile of dairy cattle at dop ≤ 50 for the first time, indicating changes in lymphocyte levels, which suggests that the animals experienced direct stress during the study.

Disorder of Biological Quality and Autophagy Process in Bovine Oocytes Exposed to Heat Stress and the Effectiveness of In Vitro Fertilization.

The main problem in dairy herds is reproductive disorders, which are influenced by many factors, including temperature. Heat stress reduces the quality of oocytes and their maturation through the influence of, e.g., mitochondrial function. Mitochondria are crucial during oocyte maturation as well as the process of fertilization and embryonic development. Disturbances related to high temperature will be increasingly observed due to global warming. In present studies, we have proven that exposure to high temperatures during the cleaving of embryos statistically significantly (at the level of p < 0.01) reduces the percentage of oocytes that cleaved and developed into blastocysts eight days after insemination. The study showed the highest percentage of embryos that underwent division in the control group (38.3 °C). The value was 88.10 ± 6.20%, while the lowest was obtained in the study group at 41.0 °C (52.32 ± 8.40%). It was also shown that high temperature has a statistically significant (p < 0.01) effect on the percentage of embryos that developed from the one-cell stage to blastocysts. The study showed that exposure to a temperature of 41.0 °C significantly reduced the percentage of embryos that split relative to the control group (38.3 °C; 88.10 ± 6.20%). Moreover, it was noted that the highest tested temperature limits the development of oocytes to the blastocyst stage by 5.00 ± 9.12% compared to controls (33.33 ± 7.10%) and cleaved embryos to blastocysts by 3.52 ± 6.80%; the control was 39.47 ± 5.40%. There was also a highly significant (p < 0.0001) effect of temperature on cytoplasmic ROS levels after 6 and 12 h IVM. The highest level of mitochondrial ROS was found in the group of oocytes after 6 h IVM at 41.0 °C and the lowest was found in the control group. In turn, at 41.0 °C after 12 h of IVM, the mitochondrial ROS level had a 2.00 fluorescent ratio, and the lowest in the group was 38.3 °C (1.08). Moreover, with increasing temperature, a decrease in the expression level of both LC3 and SIRT1 protein markers was observed. It was proved that the autophagy process was impaired as a result of high temperature. Understanding of the cellular and molecular responses of oocytes to elevated temperatures will be helpful in the development of heat resistance strategies in dairy cattle.

Ecotoxicological Assessment of Heavy Metal and Its Biochemical Effect in Fishes.

Level of toxic heavy metal concentration like lead (Pb), chromium (Cr), cadmium (Cd), iron (Fe), copper (Cu), zinc (Zn), and nickel (Ni) in thirty-six soft and hard organs and their impact on lipid profile of Hypophthalmichthys molitrix and Catla catla fish species inhibiting in Tanda Dam reservoir were investigated. The heavy metal concentrations in water, sediment, and fish of the different regions in the reservoir were determined with atomic absorption spectrophotometer. Lipid profile was carried out by AOAC official methods. The results showed that Pb was dominant among all the heavy metals in six organs, and its maximum concentration of Pb (22.5 mg kg-1 and 32.9 mg kg-1) was observed in scales in Hypophthalmichthys molitrix and tail of Catla catla, respectively. The maximum concentrations of Cd were observed in the head, scales, fins, and gills of Catla catla. The bioaccumulation of heavy metals was significantly different at (p ≤ 0.01) within the organs and between the fish species. The lipid concentration was minimum in those organs where the concentrations of heavy metals were maximum. It is clear from the findings that heavy metal accumulation reduces the lipid content of fish. It is inevitable to monitor the Tanda Dam reservoir to safeguard human health.

Evaluation of the Effect of Four Bioactive Compounds in Combination with Chemical Product against Two Spider Mites Tetranychus urticae and Eutetranychus orientalis(Acari: Tetranychidae).

Currently, pests control using chemical acaricides constitutes worries for ecologists and health care people as these chemical products create damage to the ecosystem as well as the development of spider mites resistance. Such concerns request deep and rapid feedback by looking for new alternative and eco-friendly methods. In recent years, a new field is evolving in the use of essential oils in pest management practices. Essential oils have been considered as potential pest management agents, because they demonstrate to have a broad range of bioactivity, possess contact, and fumigant toxicity. In addition, the major advantages of many plant-based acaricides lie in their low toxicity to agroecosystems. Botanical acaricides composed of essential oils may prove to be a good choice for the more persistent synthetic acaricides. In this study, the acaricidal effect of four plant-derived essential oils against adults of the two important crop pests, Tetranychus urticae (Koch) 1836 and Eutetranychus orientalis (Klein) 1936 are studied. The fumigant toxicity revealed that all the essential oils tested Mentha pulegium L., Lavandula stoechas L., Rosmarinus officinalis L., and Origanum compactum Benth (Lamiaceae family) displayed an acaricidal effect. At the highest dose (625 µl/ml), mortalities recorded were found between 91 and 98% and 92 and 99% at 24 and 48 h, respectively, for T. urticae, and between 90 and 98% and 94 and 99% at 24 and 48 h, respectively, for E. orientalis. The M. pulegium L. essential oil represents the highest activity against E. orientalis and T. urticae. For the binary combination between the EOs (essential oils) and the acaricide based on the active ingredient acequinocyl, the results showed that the mixture of O. compactum EO (essential oil) + acequinocyl exhibited an important acaricidal effect on T. urticae and E. orientalis with 99% at 24 h and 100% at 48 h of mortality, followed by M. pulegium EO + acequinocyl with 92% at 24 h and 95% at 48 h for T. urticae as well as 99% at 24 h and 100% at 48 h for E. orientalis of mortality. Whereas, the mixture of L. stoechas EO + acequinocyl presented the lowest activity against T. urticae and E. orientalis with 82-87% at 24 h and 86-90% at 48 h, respectively. The mixtures (M. pulegium EO + acequinocyl, R. officinalis EO + acequinocyl, and O. compactum EO + acequinocyl) exerted a high acaricidal effect against E. orientalis. These promising results could help to develop botanical pesticides that could be used in integrated pest management.

In Vivo Efficacy, Toxicity Assessment, and Elemental Analysis of Traditionally Used Polyherbal Recipe for Diarrhea.

A polyherbal formulation consisting of Mentha piperita L., Camellia sinensis L. Kuntze, and Elettaria cardamomum (L.) Maton with a ratio of 10 : 5 : 2, respectively, was recommended for curing nausea, vomiting, and diarrhea. Experimental validation is crucial to affirm its therapeutic property leads toward the development of modified antidiarrheal agents. This research aimed to investigate the in vivo antidiarrheal efficacy of traditionally used polyherbal recipe in a castor oil-induced animal model. Moreover, the study also presents the elemental screening and in vivo toxicity of tested polyherbal recipe. Individual plant parts of the polyherbal recipe were mixed according to the traditional prescription ratio, and hydromethanolic extract was prepared by the cold maceration process. The antidiarrheal activity was assessed by castor oil induction method, charcoal meal test, and enteropooling procedure in Sprague-Dawley rats. Elemental analysis and in vivo subacute toxicity were carried out, followed by biochemical, hematological, and histopathological analyses. Polyherbal extract significantly delayed the diarrhea onset in a dose-dependent manner and showed marked inhibition at 200 and 400 mg/kg. Fecal weight was reduced significantly (p < 0.05) at 200 mg/kg (0.26 ± 0.25) in comparison with the control (1.63 ± 0.15). The diarrhea score was zero at a concentration of 200 and 400 mg/kg. Antienteropooling effect of the extract was greater than that of loperamide. Following subacute toxicity, all the treated rats were normal, survived, and showed no changes in behavior. There were no significant differences between values of blood parameters in both the control and extract-treated groups except a significant decrease in monocytes (control 8.4; polyherbal 2.2). Elemental analysis showed a slight increase in the amount of manganese (Mn, 8.076 ppm) as compared to the WHO recommended level (2 ppm). Traditionally used polyherbal recipe is effective and safe for combating diarrheal diseases. In vivo evidence supported the use, safety, and efficacy of the polyherbal recipe that has been used as an alternative medicine for diarrhea in the study area. Inhibition of castor oil-induced diarrhea and antisecretory effect of the studied polyherbal recipe makes it a potent antidiarrheal drug without no or limited toxic effects at the tested dose after further analysis.

Reproductive Consequences of Electrolyte Disturbances in Domestic Animals.

Electrolyte balance is essential to maintain homeostasis in the body. The most crucial electrolytes are sodium (Na+), potassium (K+), magnesium (Mg2+), chloride (Cl-), and calcium (Ca2+). These ions maintain the volume of body fluids, and blood pressure, participate in muscle contractions, and nerve conduction, and are important in enzymatic reactions. The balance is mainly ensured by the kidneys, which are an important organ that regulates the volume and composition of urine, together with which excess electrolytes are excreted. They are also important in the reproductive system, where they play a key role. In the male reproductive system, electrolytes are important in acrosomal reaction and sperm motility. Sodium, calcium, magnesium, and chloride are related to sperm capacitation. Moreover, Mg2+, Ca2+, and Na+ play a key role in spermatogenesis and the maintenance of morphologically normal spermatozoa. Infertility problems are becoming more common. It is known that disturbances in the electrolyte balance lead to reproductive dysfunction. In men, there is a decrease in sperm motility, loss of sperm capacitation, and male infertility. In the female reproductive system, sodium is associated with estrogen synthesis. In the contraction and relaxation of the uterus, there is sodium, potassium, and calcium. Calcium is associated with oocyte activation. In turn, in women, changes in the composition of the follicular fluid are observed, leading to a restriction of follicular growth. Imbalance of oocyte electrolytes, resulting in a lack of oocyte activation and, consequently, infertility.

Molecular consequences of the exposure to toxic substances for the endocrine system of females.

Endocrine-disrupting chemicals (EDCs) are common in the environment and in everyday products such as cosmetics, plastic food packaging, and medicines. These substances are toxic in small doses (even in the order of micrograms) and enter the body through the skin, digestive or respiratory system. Numerous studies confirm the negative impact of EDCs on living organisms. They disrupt endocrine functions, contributing to the development of neoplastic and neurological diseases, as well as problems with the circulatory system and reproduction. EDCs affect humans and animals by modulating epigenetic processes that can lead to disturbances in gene expression or failure and even death. They also affect steroid hormones by binding to their receptors as well as interfering with synthesis and secretion of hormones. Prenatal exposure may be related to the impact of EDCs on offspring, resulting in effects of these substances on the ovaries and leading to the reduction of fertility through disturbances in the function of steroid receptors or problems with steroidogenesis and gametogenesis. Current literature indicates the need to continue research on the effects of EDCs on the female reproductive system. The aim of this review was to identify the effects of endocrine-disrupting chemicals on the female reproductive system and their genetic effects based on recent literature.

Renoprotective and Cardioprotective Potential of Moricandia sinaica (Boiss.) against Carbon Tetrachloride-Induced Toxicity in Rats.

The goal of the current study was to assess the nephroprotective and cardioprotective potential of Moricandia sinaica methanol extract (MOR-1), as well as its butanol (MOR-2) and aqueous (MOR-3) fractions against carbon tetrachloride (CCl4)-induced nephro and cardio-toxicity. Cardiac function was assessed using the biochemical parameters lactate dehydrogenase (LDH) and creatinine kinase (CK). Renal function was examined using the biochemical parameters creatinine and uric acid. The levels of nonprotein sulfhydryls (NPSH) and malondialdehyde (MDA) were used as markers of oxidative strain. A dose of 100 and 200 mg/kg of butanol fraction given prior to CCl4 treatment significantly (p < 0.05 - 0.001) protected against elevated LDH and CK levels. Similarly, treatment with silymarin (10 mg/kg) and butanol fraction (100 and 200 mg/kg) significantly (p < 0.05 - 0.001) boosted total protein levels compared to CCl4 treatment alone. The silymarin (10 mg/kg) and butanol fraction (100 and 200 mg/kg) also provided a significant (p < 0.05 - 0.001) protective effect for MDA levels. Methanol extract (MOR-1) and butanol (MOR-2) showed significant results and were recommended for further pharmacological and screening for active constituents.

Fertility Rate and Assessment of the Cytoprotective Capacity of Various Types of Holothuroidea Extracts on Spermatozoa.

For years, compounds of natural origin have been the subject of extensive biomedical research due to very interesting, new ingredients potentially useful for various pharmaceutical, medical and industrial applications. The therapeutic properties and healing benefits of sea cucumbers may result from the presence of numerous, biologically active ingredients. Sperm subjected to processing and subsequent storage at low temperatures experience a number of damage, including the loss of the integrity of the cytoplasmic membrane, DNA and acrosome defragmentation. Therefore, the aim of this experiment was to investigate the cytoprotective potential of sea cucumber extract against cryopreserved sperm and semen fertility rate. Commercially available sea cucumber extract was taken from the cellulose shell, then 790 mg of powder was weighed out and placed in 3 glass tubes containing, respectively: 10 mL of water-glycerin solution (WG), water-ethanol (EC), glycerin-ethanol (GE), glycerin-DMSO (DG). Tubes were mixed with vortex for 3 min, then placed in a water bath and incubated for 16 h at 40 °C. Six simmental bulls, 3 years old, of known health status were used for the experiment. Semen was collected from each male once a week (for 18 weeks) using an artificial vagina. After an initial assessment of semen quality, the ejaculates were pooled to eliminate individual differences between males, then diluted to a final concentration of 80 × 106 sperm/mL with a commercial extender (Optixcell, IMV, L'Aigle, France) and divided into 16 equal samples. Control (C) without additive, the test samples contained 2, 4, 6, 8 and 10 µL WG, 2, 4, 6, 8 and 10 µL WE, 2, 4, 6, 8 and 10 µL GE, 2, 4, 6, 8 and 10 µL DG. Semen was frozen/thawed and assessed for motility, viability, DNA defragmentation, mitochondrial membrane potential and acrosome integrity. It was shown a positive effect of water-glycerin (WG) and glycerine-ethanol (GE) extracts on the efficiency of sperm preservation at low temperatures. Established that, depending on the type of prepared extract, the sea cucumber can have both cytoprotective (WG, GE, WE) and cytotoxic (DG) effects. Moreover, too high concentrations of the extract can adversely affect the sperm in terms of parameters such as viability, motility, mitochondrial potential, and the integrity of the acrosome or DNA of cells. The present study, thanks to the use of model animals to study the cytoprotective potential of the sea cucumber extract, proves that it can be a potential candidate for use in semen cryopreservation technology to improve the efficiency of storage at low temperatures. Further research is needed to optimize the composition of individual types of extracts and their effect on sperm. The highest effectiveness of female fertilization was observed when doses from GE groups (2 and 4) were used for insemination. The results of this analysis prove that the addition of the tested extract may improve the fertilization efficiency.

Clinical Application of Bioextracts in Supporting the Reproductive System of Animals and Humans: Potential and Limitations.

There is an increasing demand of spices and herbs in developing countries due to the beneficial effects of plants and herbal preparations as medicines. The basic technological process of obtaining extracts from natural raw materials is extraction, consisting in etching with solvents. Plant extracts are extremely complex, multicomponent mixtures obtained from flowers, fruits, leaves, stems, twigs, or seeds of various plant materials. They are a rich source of polyphenols, flavonoids, phytosterols, carotenoids, and vitamins. The search for alternative methods of treatment is increasingly replacing the scientists' excessive focus on the healing properties of bioextracts. Recent research offers great hope for the development of alternative methods to improve the reproductive system. The use of animal models in experimental research has increased knowledge regarding the beneficial effects of bioextracts on both male and female reproductive systems and reproductive cells. Demonstrating the positive effect of plant extracts creates new opportunities for the use of biowaste, which is a by-product in various production sectors. The aim of this review is to present the functional properties of extracts of natural origin, a cross section of modern methods of their preparation, and a discussion of the possibilities of their use in the auxiliary reproductive system.

The Concentration of ProAKAP4 and Other Indicators of Cryopotential of Spermatozoa Cryopreserved in Extender with Holothuroidea Extract Addition.

The aim of this study was to determine the concentration of proAKAP4 and other indicators of cryopotential of spermatozoa cryopreserved in extender with Holothuroidea extract addition. Nine Holstein Friesian bulls, 3.5 years old, of known health status, were used for the study. The animals were kept and fed equally. Semen was collected once a week using an artificial vagina. The commercially available Holothuroidea extract was used as a supplement to the commercial extender (0, 2, 4 and 6 µL/mL) before the freezing/thawing process. The viability, motility, motion parameters, and acrosome integrity of the sperm were analyzed with (test) or without (control) extract samples. Furthermore, the concentration of the proAKAP4 biomarker in frozen sperm was assessed. It was shown that the addition of 4 and 6 µL of the extract may have a positive effect on the quality parameters of the sperm after thawing. The results indicate that extender supplementation with the above extract modulates (increases) the concentration of proAKAP4 in sperm at all tested levels. Additionally, this indicator has become helpful in identifying sperm of poor biological quality. Moreover, it has been proven that the proAKAP4 biomarker can be successfully used to evaluate the effectiveness of the use of various extenders for semen cryopreservation.

Exosomes - Spectacular role in reproduction.

Exosomes are nano-sized structures that are found in semen, epididymal -fluid, endometrium, as well as in follicular fluid. They are responsible for transporting bioactive cargo- proteins, lipids, and nucleic acids. Exosomes have been proven to influence processes in both female and male reproductive systems, including gametogenesis, acrosomal reaction, sperm capacitation, and embryo implantation in the endometrium. Exosomes are made of the same particles as the cells they come from and are secreted by normal and pathological cells. Therefore, exosomes can reflect the physiological state of cells. Moreover, due to the transportation of biomolecules, they participate in intercellular communication and can be used as biomarkers of many diseases, including ovarian, endometrial and prostate cancer. Identification of exosomes as biomarkers could contribute to a better understanding of genital dysfunction and fertility disorders.

The Role of the Natural Antioxidant Mechanism in Sperm Cells.

Molecular studies of the causes of male infertility revealed a significant contribution of oxidative stress. When excessive amounts of reactive oxygen species (ROS) are produced or antioxidant activity fails, the equilibrium between oxidation and reduction is disrupted, causing oxidative stress (OS). High levels of ROS can have an adverse effect on sperm function through the initiation of DNA damage, lipid peroxidation, loss of membrane integrity and increased permeability, inactivation of cellular enzymes, and cell apoptosis. In addition to endogenous factors such as immature sperm, leukocytes, and varicocele, potential causes of excessive ROS can also be found exogenously in males with testicular hyperthermia or exposed to environmental toxicity. To maintain the optimal functioning of sperm cells, it is, therefore, necessary to balance the redox potential, i.e., to balance ROS by antioxidants. The purpose of this review is to present the antioxidant defense systems in semen.

Disorders of the Reproductive Health of Cattle as a Response to Exposure to Toxic Metals.

The aim of this review is to comprehensively present disorders of the reproductive system in cattle exposed to contact with toxic metals. Toxic metals are a common environmental pollutant and can come from mines, smelters, fossil fuel combustion, or volcanic eruptions. Metals have the ability to bioaccumulate in living organisms, thus contaminating the food chain and may pose a threat to humans. They accumulate mainly in the liver and kidneys, but also in muscles and fat tissue. Toxic metals such as lead (Pb), arsenic (As), mercury (Hg), and cadmium (Cd) have a negative impact on the fertility of animals; they can lead to abortions, premature calving, or oocyte dysfunction. Moreover, in the male reproductive system, they disrupt spermatogenesis, and cause apoptosis of sperm and oxidative damage. The main source of exposure of livestock to toxic metals is through the consumption of feed or contaminated water. It is important to monitor the level of heavy metals in animal products to prevent human poisoning. Toxic metal biomonitoring can be performed by testing urine, blood, milk, plasma, or hair. Chromium (Cr), arsenic (As), and cadmium (Cd) are excreted in the urine, while lead can be detected by examining the blood of animals, while in milk, arsenic (As), cadmium (Cd), nickel (Ni), and lead (Pb) can be detected. Moreover, toxic metals do not biodegrade in the environment. To purify soil and waters, remediation methods, e.g., biological or chemical, should be used.

The Importance of Interferon-Tau in the Diagnosis of Pregnancy.

Several decades of improving dairy cattle towards unilateral utilization of dairy cattle led to enormous progress in the field of milk yield; however, it resulted in a number of unfavorable features, such as reproductive disorders, increased calf mortality, and reduced health. Most cases of embryo loss and/or lost pregnancies occur during the first four to five weeks of gestation; accurate detection for pregnancy during this period is likely to contribute to an improvement in gestation rates. A specific protein, interferon-tau (IFNT), stimulates interferon-stimulated genes (ISGs), and their expression increases during gestation within 21 days after insemination. In bovines, the early conceptus undergoes a phase of rapid growth and elongation before implantation, the latter occurring 2-3 weeks after fertilization. IFNT acts mainly in the endometrium of the luminal epithelium. It is a new type I interferon that regulates several genes encoding uterine-derived factors. They are crucial in the processes of preparing the uterus for placenta attachment, modifying the uterine immune system, and regulating early fetal development. Because IFNT is expressed and induces ISGs in the endometrium during pregnancy recognition, it was reasoned that surrogate markers for pregnancy or IFNT might be present in the blood and provide an indicator of pregnancy status in cattle.

The impact of regular sperm donation on bulls' seminal plasma hormonal profile and phantom response.

The aim of this study was to analyze the relationship between the concentration of hormones in the seminal plasma, the bull maintenance system in the insemination station, and the regularity of sperm donation and the response to the phantom (libido level). An additional goal was to determine whether there is a relationship between the hormonal profile in the blood, the sperm plasma, the oxidative and antioxidant profile in the blood of bulls and the biometry of their testicles and scrotum, as well as the quality of their sperm in both different seasons and intensities of reproductive use. For the study, 220 healthy and sexually mature Polish Holstein-Friesian bulls were used. They all had normal libido and were fed equally. The animals were grouped according to the scheme: young (16-20 month/n = 60) and old (26-30 month/n = 60) including: individually housed (n = 30) and group housed (n = 30) young, old individually housed (n = 30) and group housed (n = 30) (n total animals = 120); young animals donating semen once a week (every Thursday) (n = 25) and sporadically (once every two months on a random day of the week) (n = 25), old animals donating semen once a week (every Thursday) (n = 25 ) and sporadic donors (once every two months on a random day of the week) (n = 25) (n total animals = 100). When analyzing the results of this study, it should be stated that regular use has a positive effect on the secretion of sex hormones in bulls. Higher levels of testosterone and lower levels of estradiol and prostaglandins resulted in higher sexual performance, expressed by a stronger response to the phantom. The differences in favor of regular use were independent of the bull's age. The results of our research illustrate that the quality of semen and its freezing potential may depend on the season and frequency of its collection, as well as on the age of the males.

Antioxidant effect of Elamipretide on bull's sperm cells during freezing/thawing process.

BACKGROUND: Spermatozoa are subjected to drastic changes in temperature, ice crystal formation, and diverse types of stresses (chemical, physical, osmotic, and oxidative) during the cryopreservation process, which severely compromise sperm quality and fertility. In this study, we aimed to investigate the protective role of Elamipretide in the cryopreservation of bull's spermatozoa. MATERIALS AND METHODS: The study included 36 healthy Simmental bulls with an average age of 2 ± 0.5 years housed individually in pens. Two ejaculates were collected from each bull using an artificial vagina at 7 a.m. Subsequently, the semen was extended with animal protein-free commercial BIOXcell® extender (IMV Technologies) to a final concentration of 160 × 106 spermatozoa/ml, and rated in terms of motile sperm percentage, progressive motility, viability, and morphological abnormality of spermatozoa. Semen samples that showed more than 60% motility and 60% viability, were selected for the experiment. The fresh semen was then divided into five equal fractions. The first fraction was left for the control group (without Elamipretide), to the next were added in succession 0.1; 1; 5; and 10 µM of Elamipretide TFA (Trifluoroacetic) (MedChemExpress). After that semen was subjected to freezing and thawing. Next semen was assessed for motility, viability, mitochondrial membrane potential and acrosome integrity, and antioxidant activity (SOD, CAT, MDA). RESULTS: It has been shown that a concentration of 5 and 10 µM proved to be the most effective in terms of tested parameters of the quality of sperm cells subjected to cryopreservation. CONCLUSION: In conclusion, addition of the Elamipretide to the cryopreservation extender significantly improved frozen-thawed sperm cells quality and their function. The results of this study indicate that Elamipretide can be used as a cryoprotective agent to protect cells against the devastating effects of oxidative stress and increasing sperm survival after cryopreservation.

A dependency between PRL gene polymorphism and breeding parameters in Polish Black-and-White Holstein-Friesian cows.

The aim of this work was to determine the dependency between polymorphism in the PRL locus g.8398G > A and fertility rates, bodyweight of cows and calves, the course of parturition and viability of calves in Polish Black-and-White Holstein-Friesian cattle. Three genotypes (AA, AG, and GG) were identified with a frequency of 0.0293, 0.3672 and 0.6035. The frequency of the G and A alleles was respectively, 0.7871 and 0.2129. There was a significant dependency between the genetic variant in locus g.8398G > A of the PRL gene and age at first insemination, age at first calving, insemination index, length of pregnancy, length of the dry period and weight of male calves after parturition. The AA homozygotes were characterized by more favorable values of selected reproduction indices; the cows of that genotype and male calves coming from these animals were characterized by the lowest weight after parturition.