Ejaculatory Abstinence Affects the Sperm Quality in Normozoospermic Men-How Does the Seminal Bacteriome Respond?

This study was designed to describe bacterial profiles of ejaculates collected following a long and short ejaculatory abstinence set in the context of changes in the conventional, oxidative, and immunological characteristics of semen. Two specimens were collected in succession from normozoospermic men (n = 51) following 2 days and 2 h, respectively. Semen samples were processed and analyzed according to the World Health Organization (WHO) 2021 guidelines. Afterwards, sperm DNA fragmentation, mitochondrial function, levels of reactive oxygen species (ROS), total antioxidant capacity, and oxidative damage to sperm lipids and proteins were evaluated in each specimen. Selected cytokine levels were quantified using the ELISA method. Bacterial identification by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry revealed that samples collected following two days of abstinence presented with a higher bacterial load and diversity, and a greater prevalence of potentially uropathogenic bacteria including Escherichia coli, Staphylococcus aureus and Enterococcus faecalis. Only staphylococci and Escherichia coli remained present in specimens obtained after 2 h of abstinence. Whilst all samples accomplished the criteria set by WHO, a significantly higher motility (p < 0.05), membrane integrity (p < 0.05), mitochondrial membrane potential (p < 0.05), and DNA integrity (p < 0.0001) were detected following 2 h of ejaculatory abstinence. On the other hand, significantly higher ROS levels (p < 0.001), protein oxidation (p < 0.001), and lipid peroxidation (p < 0.01) accompanied by significantly higher concentrations of tumor necrosis factor alpha (p < 0.05), interleukin-6 (p < 0.01), and interferon gamma (p < 0.05) were observed in specimens collected after two days of abstinence. It may be summarized that shorter ejaculatory abstinence does not compromise sperm quality in normozoospermic men, while it contributes to a decreased occurrence of bacteria in semen which is accompanied by a lower probability of damage to spermatozoa by ROS or pro-inflammatory cytokines.

Mercury in scarletina bolete mushroom (Neoboletus luridiformis): Intake, spatial distribution in the fruiting body, accumulation ability and health risk assessment.

In the present work, we focused on two aspects of mercury (Hg) bioconcentration in the above-ground parts of Neoboletus luridiformis. In the first part, we monitored the bioconcentration potential of individual anatomical parts of a particular fruiting body and evaluated the obtained data by the spline interpolation method. In the second part, we focused on assessing the mercury content in 378 samples of N. luridiformis and associated samples of substrates from 38 localities with different levels of Hg content in Slovakia. From the obtained data of Hg content in samples of substrate and fungi, we evaluated ecological indicators (geoaccumulation index - Igeo, contamination factor - Cf a potential ecological risk - PER), bioconcentration indicators (bioconcentration factor - BCF; cap/stipe quotient - Qc/s) and health indicators (percentage of provisional tolerable weekly intake - %PTWI a target hazard quotient - THQ). Based on the Hg distribution results, the highest Hg content was found in the tubes & pores (3.86 mg/kg DW), followed by the flesh of cap (1.82 mg/kg DW). The lowest Hg content was in the stipe (1.23 mg/kg DW). The results of the BCF values indicate that the studied species can be included in the category of mercury accumulators. The results of the ecological indices representing the state of soil pollution pointed out that two localities (Malachov and Nizná Slaná) stood apart from all monitored localities and showed a state of an extremely disturbed environment. This fact was also reflected in the values of Hg content in the fruiting bodies of the studied mushroom species. In the case of the consumption of mushrooms from these localities, it can be stated that long-term and regular consumption could have a negative non-carcinogenic effect on the health of consumers. It was confirmed by the %PTWI (Malachov: 57.8%; Nizná Slaná: 53.2%) and THQ (Malachov: 1.11 Nizná Slaná: 1.02). The locality Cacín-Jelsovec is interesting from the bioconcentration characteristics point of view, where the level of environmental pollution was the lowest (Hg content in the soil was below the background value) compared to other localities, however, the THQ value was the highest (1.29).

The effect of brown seaweed and polyphenol supplementation in male rabbits on the blood profile and antioxidant markers.

Currently, in animal nutrition, the replacement of synthetic substances with natural ones was expected to improve animal health. The aim of the present study was to evaluate the effects of a dietary brown seaweed and plant polyphenol mixture in adult male rabbits on the haematological profile and antioxidant markers. Twenty-four adult male rabbits were divided into three experimental groups receiving a control diet (C) or diets supplemented with 0.3% (T1) and 0.6% (T2) of a feed additive containing brown seaweed (Laminaria spp.) and plant extracts of seaweed origin. The trial lasted for 90 days. A lower potassium concentration was observed at 30 days in the T2 group, compared with the T1 and C groups. An increase in the antioxidant status was observed (P < 0.05) from day 60 of the trial in the rabbits fed diets with an algae-polyphenolic supplement (T1 and T2 groups). Concluding, the diet supplementation of brown seaweed and polyphenol stimulates the antioxidant status of the blood, however, it does not affect the haematological profile.

Consumption of bitter apricot seeds affects lipid and endocrine profile in women.

Natural products have been attracting increasing attention in human diet, both due to the possible negative effects of synthetic food additives on human health and the increased consumer perception. Apricot seeds contain a wide variety of bioactive components and their consumption is associated with a reduced risk of chronic diseases. The objective of the present study was to evaluate the effect of consumption of bitter apricot seeds on blood lipid and endocrine profile in Slovak women (n = 18, 41.60 ± 11.28 years) of reproductive age. Volunteers consumed 60 mg.kg-1 of body weight of bitter apricot seeds divided into 8-12 doses daily for 42 days. During the experiment, three blood collections were carried out (at the beginning of the experiment - day 0, and after 21 and 42 days of consumption apricot seeds). Lipid profile was measured in terms of - total cholesterol (T-C, enzymatic photometric method), low-density cholesterol (LDL-C, calculated using the Friedewald equation), high-density cholesterol (HDL-C, direct clearance method), triglycerides (TG, enzymatic colorimetric method) whereas endocrine profile - follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), progesterone (P4), 17ß-estradiol (E2), testosterone, and androstenedione was assessed by ELISA. The blood levels of T-C, HDL-C and T-C did not change significantly (P > 0.05), however, the level of LDL-C decreased significantly (P < 0.05) after 42 days. On the other hand, there was a significant (P < 0.05) increase of T-C and TG after 21 days. The blood level of FSH, testosterone and androstenedione increased significantly (P < 0.05) although the levels of LH, PRL, P4 and E2 did not change (P > 0.05) after 42 days. The level of PRL and testosterone significantly (P < 0.05) increased and E2 significantly decreased after 21 days of apricot seeds consumption. The study suggests that daily consumption of apricot seeds may affect plasma lipid and endocrine profile in women of reproductive age.

Potential toxicity of cyanogenic glycoside amygdalin and bitter apricot seed in rabbits-Health status evaluation.

Amygdalin is one of the most studied secondary metabolites of Prunus genus. It is a cyanogenic glycoside which was initially obtained from the bitter almonds seeds and is a major component of the seeds of plants, such as apricots, almonds, peaches, apples and other rosaceous plants. The views of scientists on the use of amygdalin have been contradictory for many years, partly because toxicokinetics and metabolism of amygdalin still have not been adequately explored. The present in vivo study was designed to reveal whether pure amygdalin intramuscularly injected or apricot seeds oral consumption induce changes in overall health status of rabbit as a biological model. A total of 60 adult rabbits were randomly divided into five groups. The control group received no amygdalin while the two experimental groups E1 and E2 received a daily intramuscular injection of amygdalin at doses 0.6 and 3.0 mg/kg bw. The experimental groups E3 and E4 were fed crushed bitter apricot seeds (Prunus armeniaca L.), at doses 60 and 300 mg/kg bw, mixed with commercial feed for rabbits. Blood collection was carried out after 14 days. Biochemical, haematological and antioxidant enzymes activity analysis were performed and statistically evaluated. A short-term amygdalin administration had negligible impact on biochemical parameters-mainly level of urea, bilirubin, cholesterol. Haematological profile of rabbits was influenced very slightly-non-significant platelet count and platelet percentage increase, erythrocytes count and haemoglobin decrease. SOD activity of rabbits decreased significantly (p > 0.05) after apricot seeds consumption (102.3 U/ml) in comparison to control (117.4 U/ml). Differences might be connected to diverse metabolism by different administration routes and at the same time by the presence of other substances in apricot seeds (phytosterols, polyphenols, fatty acids). However, a short-term consumption had only slight effect on health status of rabbits and at recommended doses did not represent risk for their health.

Parallel effect of 4-octylphenol and cyclic adenosine monophosphate (cAMP) alters steroidogenesis, cell viability and ROS production in mice Leydig cells.

Over the last decade, there is growing incidence of male reproductive malfunctions. It has been documented that numerous environmental contaminants, such as endocrine disruptors (EDs) may adversely affect the reproductive functions of humans as well as wildlife species. The aim of this in vitro study was to examine the effects of 4-octylphenol (4-OP) on the steroidogenesis in mice Leydig cells. We evaluated the impact of this endocrine disruptor on the cholesterol levels and hormone secretion in a primary culture. Subsequently, we determined the cell viability and generation of reactive oxygen species (ROS) following 4-OP treatment. Isolated mice Leydig cells were cultured in the presence of different 4-OP concentrations (0.04-5.0 µg/mL) and 1 mM cyclic adenosine-monophosphate during 44 h. Cholesterol levels were determined from the culture medium using photometry. Quantification of steroid secretion was performed by enzyme-linked immunosorbent assay. The cell viability was assessed using the metabolic activity assay, while ROS production was assessed by the chemiluminescence technique. Slightly increased cholesterol levels were recorded following exposure to the whole applied range of 4-OP, without significant changes (P>0.05). In contrast, the secretion of steroid hormones, specifically dehydroepiandrosterone, androstenedione, and testosterone was decreased following exposure to 4-OP. Experimental doses of 4-OP did not affect cell viability significantly; however a moderate decrease was recorded following the higher doses (2.5 and 5.0 µg/mL) of 4-OP. Furthermore, relative treatment of 4-OP (5.0 µg/mL) caused a significant (P < 0.001) ROS overproduction in the exposed cells.

Resveratrol offers protection to oxidative stress induced by ferrous ascorbate in bovine spermatozoa.

Resveratrol (RES) is a natural polyphenol and phytoestrogen exhibiting cardioprotective, anticancer, antibacterial and vasorelaxing properties. It is also a powerful reactive oxygen species (ROS) scavenger and chelating agent. This study was designed to determine the efficiency of RES to reverse the ROS-mediated impairment of the motility, viability and intracellular antioxidant profile of bovine spermatozoa. Spermatozoa were washed out of fresh bovine semen, suspended in 2.9% sodium citrate and subjected to RES treatment (5, 10, 25 and 50 µmol L(-1)) in the presence or absence of a pro-oxidant, i.e., ferrous ascorbate (FeAA; 150 µmol L(-1) FeSO4 and 750 µmol L(-1) ascorbic acid) during a 6-h in vitro culture. Spermatozoa motion parameters were assessed using the SpermVision computer-aided sperm analysis (CASA) system. Cell viability was examined with the metabolic activity (MTT) assay, and the nitroblue-tetrazolium (NBT) test was applied to quantify the intracellular superoxide formation. Cell lysates were prepared at the end of the in vitro experiments in order to investigate the intracellular activity of superoxide dismutase (SOD), catalase (CAT), as well as the concentrations of glutathione (GSH) and malondialdehyde (MDA). FeAA treatment led to a reduced sperm motility (P < 0.001) and viability (P < 0.001), decreased the antioxidant parameters of the samples (P < 0.001 in case of SOD; P < 0.01 with respect to CAT; P < 0.05 in relation to GSH) but increased the superoxide production (P < 0.001) and lipid peroxidation (P < 0.001). RES supplementation resulted in a preservation of the spermatozoa vitality and antioxidant characteristics (P < 0.001 in case of SOD; P < 0.01 with respect to 25-50 µmol L(-1) RES and P < 0.05 in relation to 10 µmol L(-1) RES; P < 0.05 in case of GSH), with 50 µmol L(-1) RES proving to be the most effective RES concentration. Our results suggest that RES possesses significant antioxidant properties that may prevent the deleterious effects caused by ROS to spermatozoa, and preserve the fertilization potential of male reproductive cells.

Blood concentration of copper, cadmium, zinc and lead in horses and its relation to hematological and biochemical parameters.

Environmental pollution results in serious health hazards to animals and blood analysis serves as a good alternative for health status assessment. The target of this study was to analyze the concentration of selected metals in equine blood, to analyze the blood parameters and to find possible correlations. Blood samples were collected from the vena jugularis of healthy adult horses. The highest concentration of all elements was found in whole blood (Cu 3.84 ± 0.90 mg L(-1); Cd = 0.81 ± 0.90 mg L(-1); Zn 26.67 ± 14.12 mg L(-1); Pb 9.33 ± 5.76 mg L(-1)). Higher concentrations of copper, cadmium, zinc and lead were detected in blood clots compared to blood sera (44.04%). A similar tendency was found for cadmium (50%), zinc (13.08%) and lead (46.02%), which showed generally higher concentrations in blood clots (cells). Correlation analysis proved some relations between analyzed elements. In blood clots there is a strong positive correlation between Cd - Pb (r = 0.93) and Zn - Pb (r = 0.71) was detected. For biochemical and hematological parameters mainly medium correlations were detected. Obtained results prove different correlations of analyzed elements in blood components as well as the effect on parameters of blood biochemical and hematological profiles.

Nickel-induced structural and functional alterations in porcine granulosa cells in vitro.

The present study was aimed at investigating the effect of nickel chloride (NiCl2) on secretion of progesterone (P), ultrastructure and apoptosis in porcine granulosa cells. NiCl2 was added to the cells to achieve a Ni(2+) concentration of 62.5, 125, 250, 500 and 1,000 µmol/L. A control group contained no NiCl2 addition. Quantification of P was performed directly from aliquots of the media from control and treated porcine granulosa cells after 48 h of culture using radioimmunoassay. Quantification of apoptotic cells was performed using terminal deoxynucleotidyl transferase dUTP nick end labelling assay, and ultrastructural changes were analyzed using transmission electron microscopy. A concentration-dependent depletion of P production was observed significantly for 1,000 µmol/L NiCl2. The percentage of apoptotic cells was increased in all experimental groups significantly only after addition of 1,000 µmol/L NiCl2. After addition of ≥250 µmol/L NiCl2, a higher incidence of euchromatin was observed. Also, lipid droplets and vacuoles in the cytoplasm increased after addition of ≥250 µmol/L NiCl2. NiCl2 induced the decrease in numbers of mitochondria and smooth endoplasmic reticulum after treatment with ≥500 µmol/L NiCl2. Our findings suggest a negative effect of NiCl2 on steroidogenesis and apoptosis as well as ultrastructure of porcine granulosa cells.