Microelements, Fatty Acid Profile, and Selected Biomarkers in Grass Carp (Ctenopharyngodon idella) Muscle Tissue: Seasonal Variations and Health Risk Assessment.

The study assesses associations between microelement levels, fatty acid composition, and oxidative stress markers in grass carp muscle in the summer and autumn seasons. Additionally, various factors were considered, including the estimated daily intake (EDI), target hazard quotient (THQ), total THQ (TTHQ), and metal pollution index (MPI), to evaluate potential health risks for consumers. The microelements (Al, As, Ba, Cd, Co, Cr, Cu, Fe, Hg, Li, Mn, Ni, Pb, Se, Sr, and Zn) were determined using inductively coupled plasma optical emission spectrometry (ICP-OES), and total mercury was determined using cold-vapor atomic absorption spectroscopy (CV-AAS). Fatty acid profiling was realized using gas chromatography (GC) detection with a flame ionization detector (FID). The overall tendency of microelement levels was as follows: Fe > Zn > Al > Sr > Ba > Ni > Se > Cr> Cu > Mn > Pb > As > Li > Hg;

Blood Concentration of Macro- and Microelements in Women Who Are Overweight/Obesity and Their Associations with Serum Biochemistry.

Risk elements in blood matrices can affect human health status through associations with biomarkers at multiple levels. The aim of this study was to analyze 15 macro- and microelements in the blood serum of women with overweight (BMI of ≥25 kg/m2) and obesity (BMI of ≥30 kg/m2) and to examine possible associations with biochemical, liver enzymatic parameters, and markers of oxidative stress. Based on the power calculation, the study involved women (in the postmenopausal stage) with overweight (n = 26) and obesity (n = 22), aged between 50-65 years. Multifrequency bioelectrical impedance analysis was used to measure body composition parameters. Concentrations of elements were determined by inductively coupled plasma optical emission spectrometry, and Hg was measured using cold-vapor atomic absorption spectroscopy. Individuals with obesity, as indicated by a higher BMI, percentage of body fat, and visceral fat area, had elevated serum levels of Ca, Mg, Fe, Al, Sr, Pb, and Hg. Concentrations of Al, Cu, K, Sb, Zn, and Pb significantly affected biochemical and liver function markers in women with overweight or obesity. Elements such as Cu and Al were associated with increased total cholesterol. The correlation analysis between total antioxidant status and Cu, Al, and Ni confirmed associations in both groups. Our findings underscore the importance of addressing excess body weight and obesity in relation to risk elements. The results of the research could be beneficial in identifying potential targets for the treatment or prevention of comorbidities in people with obesity.

Seasonal assessment of selected trace elements in grass carp (Ctenopharyngodon idella) blood and their effects on the biochemistry and oxidative stress markers.

Environmental pollution by anthropogenic activity is still a highly relevant global problem. Aquatic animals are a specifically endangered group of organisms due to their continuous direct contact with the contaminated environment. Concentrations of selected trace elements in the grass carp (Ctenopharyngodon idella) (n = 36) blood serum/clot were monitored. Possible effects of the elements on selected biochemical and oxidative markers were evaluated. The concentrations of trace elements (Al, Ba, Be, Bi, Cd, Co, Cr, Cu, Fe, Ga, Mn, Mo, Ni, Pb, Sr, Tl, and Zn) were analysed in the fish blood serum and blood clot by inductively coupled plasma optical emission spectrometry (ICP OES). A general scheme of decreasing concentrations of trace elements in the blood serum samples was: Zn ˃ Fe ˃ Sr ˃ Ba ˃ Ni ˃ Al ˃ Cu ˃ Be ˃ Co; < LOQ (below limit of quantification): Bi, Cd, Cr, Ga, Mn, Mo, Pb, Tl; and in the case of the blood clot, the scheme was as follows: Fe ˃ Zn ˃ Sr ˃ Al ˃ Ni ˃ Ba ˃ Cu ˃ Be ˃ Co ˃ Mn; < LOQ (below limit of quantification): Bi, Cd, Cr, Ga, Mo, Pb, Tl. Significant differences among the seasons were detected. The Spearman R correlation coefficients and linear or non-linear regression were used to evaluate direct relationships between trace elements and selected blood biomarkers. The correlation analysis between biochemical parameters (Na, K, P, Mg, AST, ALT, ALP, GGT, TAG, TP, urea, glucose) and trace elements (Al, Ba, Be, Cu, Fe, Ni, Sr, and Zn) concentrations confirmed statistically significant interactions in both seasons (summer and autumn). The regression analysis between oxidative stress markers (ROS, GPx, creatinine, uric acid, and bilirubin) and elements (Al, Ba, Co, Cu, Fe, Ni, and Sr) content confirmed statistically significant interactions. The results point to numerous connections between the observed elements and the physiological parameters of freshwater fish.

Screening of Toxic Effects of Neonicotinoid Insecticides with a Focus on Acetamiprid: A Review.

Recently, neonicotinoids have become the fastest-growing class of insecticides in conventional crop protection, with extensive usage against a wide range of sucking and chewing pests. Neonicotinoids are widely used due to their high toxicity to invertebrates, simplicity, flexibility with which they may be applied, and lengthy persistence, and their systemic nature ensures that they spread to all sections of the target crop. However, these properties raise the risk of environmental contaminations and potential toxicity to non-target organisms. Acetamiprid is a new generation insecticide, which is a safer alternative for controlling insect pests because of its low toxicity to honeybees. Acetamiprid is intended to target nicotinic acetylcholine receptors in insects, but its widespread usage has resulted in negative impacts on non-target animals such as mammals. This review summarizes in vivo and in vitro animal studies that investigated the toxicity of specific neonicotinoids. With summarized data, it can be presumed that certain concentrations of neonicotinoids in the reproductive system cause oxidative stress in the testis; spermatogenesis disruption; spermatozoa degradation; interruptions to endocrine function and Sertoli and Leydig cell function. In the female reproductive system, acetamiprid evokes pathomorphological alterations in follicles, along with metabolic changes in the ovaries.

Biogenic Elements and Heavy Metals in Hermann's Tortoises-Testudo hermanni: Effect on Serum Biochemistry and Oxidative Status Parameters.

BACKGROUND: Conservation of species diversity is the need of the hour for preserving life forms on Earth. Extinction of any part of the ecosystem has negative impacts on many processes and systems. The objective of this work was to analyze some biochemical and molecular indicators and their correlations to biogenic elements and heavy metals in Testudo hermanni (n = 16). METHODS: Biochemical parameters were analyzed using the commercial kit DiaSys and biochemical analyzer Randox RX Monza. Sodium, potassium, and chlorides were measured using the EasyLite analyzer. Oxidative stress was evaluated using colorimetric and luminometric methods. Quantification of chemical elements in the blood was carried out using inductively coupled plasma mass spectrometry (ICPS). RESULTS: Biochemical values of analyzed samples from Hermann's tortoises were almost the same as referential values described by multiple authors, with minor aberrations in the total protein parameter. Values of arsenic (As) and nickel (Ni) showed correlation with biochemical parameters and the parameters of oxidative stress. Cadmium (Cd) exhibited correlation with aspartate aminotransferase (AST). CONCLUSIONS: This study reports correlations among four heavy metals, and their levels were again correlated with biochemical and molecular parameters in Hermann's tortoises.

MALDI-TOF MS Indirect Beta-Lactamase Detection in Ampicillin-Resistant Haemophilus influenzae.

Rapid identification of beta-lactamase-producing strains of Haemophilus influenzae plays key role in diagnostics in clinical microbiology. Therefore, the aim of this study was the rapid determination of beta-lactamase's presence in H. influenzae isolates via indirect detection of degradation ampicillin products using MALDI-TOF MS. H. influenzae isolates were subjected to antibiotic resistance testing using disk diffusion and MIC methodologies. Beta-lactamase activity was tested using MALDI-TOF MS, and results were compared to spectral analysis of alkaline hydrolysis. Resistant and susceptible strains of H. influenzae were distinguished, and strains with a high MIC level were identified as beta-lactamase-producing. Results indicate that MALDI-TOF mass spectrometry is also suitable for the rapid identification of beta-lactamase-producing H. influenzae. This observation and confirmation can accelerate identification of beta-lactamase strains of H. influenzae in clinical microbiology, which can have an impact on health in general.

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.

Microbiota, Phagocytic Activity, Biochemical Parameters and Parasite Control in Horses with Application of Autochthonous, Bacteriocin-Producing, Probiotic Strain Enterococcus faecium EF 412.

The beneficial influence of bacteriocin-producing, probiotic, mostly non-autochthonous bacteria has already been reported in various animals. However, their use in horses provides limited information, and results with autochthonous bacteria have not been reported. Therefore, the main objective of this model study was to test the effect of autochthonous, bacteriocin-producing faecal strain Enterococcus faecium EF 412 application in horses. One gram of freeze-dried EF 412 strain (109 CFU/mL for 21 days) was applied to horses in a small feed ball. Clinically healthy horses (12), Slovak warm-blood breed of various ages (5-13 years), were involved in a 35-day-long experiment, also functioning as control for themselves. They were stabled in separate boxes (university property), fed twice a day (hay, whole oats or grazed) with water access ad libitum. Sampling was performed at the start of the experiment, i.e. at days 0/1, 21 (3 weeks of EF 412 application) and at day 35 (2 weeks of EF 412 cessation). EF 412 colonized GIT of horses was 3.54 ± 0.75 CFU/g (log 10) at day 21. The eggs of the nematode Strongylus spp. were not found in horses after EF 412 application, and Eimeria spp. oocysts were similarly not found. The other microbiota were not reduced as evaluated by the use of standard method. Using next-generation sequencing, at phylum level, phyla Bacteroidetes and Firmicutes dominated and at family level, they were Bacteroidales BS11 and S24-7 gut goups and Lentisphaerae. In horses, the increasing tendency in phagocytic activity was noted after EF 412 application. Biochemical parameters were in the physiological range. Total protein value was significantly decreased at day 21 compared with day 0/1 as well as with day 35 (P < 0.05). Cholesterol and triglycerides were influenced (decreased) at day 21 compared with day 0/1 and day 35. Neither nematode eggs Strongylus spp. nor Eimeria spp. oocysts were found in faeces after EF 412 application. Autochthonous, faecal strain E. faecium EF 412 showed promising application potential.

Use of MALDI-TOF MS to Discriminate between Aflatoxin B1-Producing and Non-Producing Strains of Aspergillus flavus.

Aflatoxin B1 (AFB1) is one of the most toxic mycotoxins. One of the producers of AFB1 is Aspergillus flavus. Therefore, its rapid identification plays a key role in various sectors of the food and feed industry. MALDI-TOF mass spectrometry is one of the fastest and most accurate methods today. Therefore, the aim of this research was to develop the rapid identification of producing and non-producing strains of A. flavus based on the entire mass spectrum. To accomplish the main goal a different confirmatory MALDI-TOF MS and TLC procedures such as direct AFB1 identification by scraping from TLC plates, A. flavus mycelium, nutrient media around A. flavus growth, and finally direct AFB1 identification from infected wheat and barley grains had to be conducted. In this experiment, MALDI-TOF mass spectrometry with various modifications was the main supporting technology. All confirmatory methods confirmed the presence of AFB1 in the samples of aflatoxin-producing strains of A. flavus and vice versa; AFB1 was not detected in the case of non-producing strains. Entire mass spectra (from 2 to 20 kDa) of aflatoxin-producing and non-producing A. flavus strains were collected, statistically analyzed and clustered. An in-depth analysis of the obtained entire mass spectra showed differences between AFB1-producing and non-producing strains of A. flavus. Statistical and cluster analysis divided AFB1-producing and non-producing strains of A. flavus into two monasteries. The results indicate that it is possible to distinguish between AFB1 producers and non-producers by comparing the entire mass spectra using MALDI-TOF MS. Finally, we demonstrated that if there are established local AFB1-producing and non-producing strains of A. flavus, the entire mass spectrum database identification of aflatoxigenic A. flavus strains can be even faster and cheaper, without the need to identify the toxin itself.

Exceptional Properties of Lepidium sativum L. Extract and Its Impact on Cell Viability, Ros Production, Steroidogenesis, and Intracellular Communication in Mice Leydig Cells In Vitro.

The prevalence of reproductive dysfunction in males has risen in the last few years, and alternative therapies are gradually gaining in popularity. Our in vitro study aimed to evaluate the potential impact of Lepidium sativum L. on mice TM3 Leydig cells, concerning basal parameters such as cell viability, cell membrane integrity, and lysosomal activity, after 24 h and 48 h exposure. Moreover, reactive oxygens species generation, sex-steroid hormone secretion, and intercellular communication were quantified. In the present study, the microgreen extract from Lepidium was rich in ferulic acid, 4-OH benzoic acid, and resveratrol, with a significant antioxidant activity. The results showed that lower experimental doses (62.5-250 µg/mL) could positively affect the observed parameters, with significant differences at 250 µg/mL after 24 h and 48 h, respectively. Potential risks could be associated with higher concentrations, starting at 500 µg/mL, 1000 µg/mL, and 2000 µg/mL of Lepidium. Nevertheless, biochemical quantification indicated a significant antioxidant potential and a rich content of biologically active molecules at the applied doses, and time determined the intracellular response of the cultured model.

Phytonutrients of Bitter Apricot Seeds Modulate Human Lipid Profile and LDL Subfractions in Adults with Elevated Cholesterol Levels.

The objective of the present study was to evaluate the effect of short-term consumption of bitter apricot seeds phytonutrients on cardiovascular risk factors with a special focus on LDL cholesterol subfractions using the Lipoprint system. A group of 34 adult volunteers (21 female/13 male) consumed 60 mg kg-1 of body weight of bitter apricot seeds daily for 42 days. Subjects were divided into two groups: one with normal cholesterol levels (NTC) and one with elevated total cholesterol levels (ETC). Blood serum levels of total cholesterol (T-C), low-density cholesterol (LDL-C), high-density cholesterol (HDL-C), and triglycerides (TG) did not change significantly (p > 0.05) in NTC group. However, there were significant decreasing of T-C (p ˂ 0.05) and LDL-C (p < 0.01) in ETC group. The LDL1, LDL2, and atherogenic LDL3-7 subfractions progressively decreased after 42 days of apricot seeds consumption in ETC group (p < 0.05). Apricot seeds consumption was associated with a significant increase in the mean LDL particle size especially in ETC group (p ˂ 0.01). The results of the present study support the hypothesis that daily consumption of bitter apricot seeds for 42 days positively modified the lipoprotein profile in the group with elevated total cholesterol.

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).

In vivo effects of aflatoxin B1 and benzo[a]pyrene on the heart muscle of chicken embryos.

The purpose of the present study was to evaluate the effects of aflatoxin B1 (AFB1) and benzo[a]pyrene (BaP) on the heart muscle of chicken embryos of the broiler line Ross 308. The benzo[a]pyrene in the organic oil solution was injected in ovo on the 6th day of the incubation in doses of: 0.1, 0.5, and 1 mg/kg weight of eggs; the aflatoxin B1 in the organic oil solution was injected in ovo on the 6th day of the incubation into the yolk in doses of 80, 120 and 240 ng/kg weight of eggs. Multiple biochemical and hepatic parameters have been observed, including sodium, potassium, chloride, cholesterol, uric acid, total proteins, aminotransferase aspartate, and aminotransferase alanine. A low dose of AFB1 and BaP administered in ovo during early embryonic development had a significant impact on chicken embryonic development, as demonstrated by alterations in biochemical, mineral, and hepatic parameters.

Carbapenemase Producing Klebsiella pneumoniae (KPC): What Is the Best MALDI-TOF MS Detection Method.

Klebsiella pneumoniae carbapenemase (KPC)-producing bacteria is a group of highly dangerous antibiotic resistant Gram-negative Enterobacteriaceae. They cause infections associated with significant morbidity and mortality. Therefore, the rapid detection of KPC-producing bacteria plays a key role in clinical microbiology. Matrix assisted laser desorption/ionization time-of- flight (MALDI-TOF) is a rapidly evolving technology that finds application in various clinical, scientific, and industrial disciplines. In the present study, we demonstrated three different procedures of carbapenemase-producing K. pneumoniae (KPC) detection. The most basic model of MALDI-TOF instrument MS Microflex LT was used, operating in the linear ion-positive mode, commonly used in modern clinical laboratories. The first procedure was based on indirect monitoring of carbapenemase production with direct detection of hydrolyzed carbapenem antibiotic degradation products in the mass spectrum. The second procedure was based on direct detection of blaKPC accompanying peak with an 11,109 Da in the mass spectrum of carbapenemase-producing K. pneumoniae (KPC), which represents the cleaved protein (pKpQIL_p019) expressed by pKpQIL plasmid. In addition, several unique peaks were detected in the carbapenemase-producing K. pneumoniae (KPC) mass spectrum. The third procedure was the identification of carbapenemase-producing K. pneumoniae (KPC) based on the protein fingerprint using local database created from the whole mass spectra. By comparing detection procedures, we determined that the third procedure was very fast and relatively easy. However, it requires previous verification of carbapenemase-producing K. pneumoniae (KPC) using other methods as genetic blaKPC identification, detection of carbapenem degradation products, and accompanying peak with 11,109 Da, which represents cleaved pKpQIL_p019 protein expressed by pKpQIL plasmid. Detection of carbapenemase-producing K. pneumoniae using MALDI-TOF provides fast and accurate results that may help to reduce morbidity and mortality in hospital setting when applied in diagnostic situations.

Composition of Stallion Seminal Plasma and Its Impact on Oxidative Stress Markers and Spermatozoa Quality.

The composition of seminal plasma of individual sires varies and so does the fertilizing ability. Micro and macro elements along with seminal enzymes, hormones, proteins, and lipids contained in seminal plasma are essential for the proper physiological function of spermatozoa. However, elevated levels against the normal physiological values, especially in the case of trace metals, result in the production of reactive oxygen species. The deficiency of antioxidants in the seminal plasma that could scavenge free radicals causes an impairment of spermatozoa quality. Ejaculates were obtained from 19 stallions. The fresh semen was analyzed to evaluate qualitative parameters of spermatozoa in terms of the motility, viability, and integrity of DNA. Separated seminal plasma underwent the assessment of the chemical and biochemical composition and RedOx markers. Based on the obtained concentrations of individual chemical elements, the correlation analysis suggested a negative impact of Cu in seminal plasma on the SOD, GPx, and LPO. Contrary, positive correlation was detected between FRAP and motility features. While Cu negatively correlated with sperm motion parameters, the adverse effect on viability was suggested for Cd. Our data suggest that seminal plasma has a potential due to its availability to become the potential biomarker of the reproductive health of farm animals.

Antifungal and synergistic activities of some selected essential oils on the growth of significant indoor fungi of the genus Aspergillus.

The study aimed to assess the antifungal activity of twenty-five essential oils (EOs) and the potential synergistic activity of the most effective EOs against significant indoor fungi of the genus Aspergillus [A. fumigatus (KBio-122), A. flavus (KBio-134), A. terreus (KBio-145) and A. niger (KBio-202)]. The chemical composition of all EOs was evaluated by the gas chromatography coupled with mass spectrometry (GC/MS) and gas chromatography with flame ionization detector (GC-FID) analysis. The antifungal susceptibility of EOs was evaluated by using the broth microdilution method. The most effective EOs were selected to determine the minimum inhibitory concentrations (MICs) and minimum fungicidal concentrations (MFCs) at a concentration range from 256 to 0.125 µg/mL. For the synergistic activities, the most effective EOs were tested using the chessboard pattern. The most sensitive strain to treatments with essential oils alone and in the combination of EOs was A. flavus (KBio-134). The chessboard assay showed that combinations of lemongrass and thyme EOs proved the most potent synergistic antifungal activity (FICI = 0.1875) against A. fumigatus (KBio-122). The synergy displayed by a combination of some EOs may be used to control fungal growth or increasing resistance to available synthetic antifungals, consequently permitting the reduction of their most active doses.

In vitro effect of ferrous sulphate on bovine spermatozoa motility parameters, viability and Annexin V-labeled membrane changes.

The aim of this study was to assess the dose- and time-dependent in vitro effects of ferrous sulphate (FeSO4.7H2O) on the motility parameters, viability, structural and functional activity of bovine spermatozoa. Spermatozoa motility parameters were determined after exposure to concentrations (3.90, 7.80, 15.60, 31.20, 62.50, 125, 250, 500 and 1000 µM) of FeSO4.7H2O using the SpermVisionTM CASA (Computer Assisted Semen Analyzer) system in different time periods. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) assay, and the Annexin V-Fluos was applied to detect the membrane integrity of spermatozoa. The initial spermatozoa motility showed increased average values at all experimental concentrations compared to the control group (culture medium without FeSO4.7H2O). After 2 h, FeSO4.7H2O stimulated the overall percentage of spermatozoa motility at the concentrations of ≤ 125 µM. However, experimental administration of 250 µM of FeSO4.7H2O significantly (P < 0.001) decreased the spermatozoa motility but had no negative effect on the cell viability (P < 0.05) (Time 2 h). The lowest viability was noted after the addition of ≥ 500 µM of FeSO4.7H2O (P < 0.001). The concentrations of ≤ 62.50 µM of FeSO4.7H2O markedly stimulated (P < 0.001) spermatozoa activity after 24 h of exposure, while at high concentrations of ≥ 500 µM of FeSO4.7H2O the overall percentage of spermatozoa motility was significantly inhibited (P < 0.001) and it elicited cytotoxic action. Fluorescence analysis confirmed that spermatozoa incubated with higher concentrations (≥ 500 µM) of FeSO4.7H2O displayed apoptotic changes, as detected in head membrane (acrosomal part) and mitochondrial portion of spermatozoa. Moreover, the highest concentration and the longest time of exposure (1000 µM of FeSO4.7H2O; Time 6 h) induced even necrotic alterations to spermatozoa. These results suggest that high concentrations of FeSO4.7H2O are able to induce toxic effects on the structure and function of spermatozoa, while low concentrations may have the positive effect on the fertilization potential of spermatozoa.

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.

Cytotoxic effect of aminoglycoside antibiotics on the mammalian cell lines.

Aminoglycoside antibiotics have been used for treating serious but also routine infections in veterinary and human medicine for many years. The basic aim of this work is to evaluate the cytotoxicity of dihydrostreptomycin and neomycin in vitro on three cell cultures - BHK-21 (Syrian golden hamster kidney fibroblast), VERO (African green monkey kidney fibroblast) and FEA (feline embryonic fibroblast) cells. The morphological changes were examined by Giemsa staining. Cells were dried and visualized under fluorescence microscope. After the exposure to different experimental doses of dihydrostreptomycin (812.5-20000 µg/mL) and neomycin (1000-20000 µg/mL) during 24 h, the viability of BHK-21, FEA and VERO cell lines were evaluated by MTT assay. Viability of BHK-21 cells significantly (P < 0.001) decreased after treatment with 3500; 5500 and 7500 µg/mL of dihydrostreptomycin and 9000; 10000 and 20000 µg/mL of neomycin. The FEA cell viability decreased significantly (P < 0.001; P < 0.01) at 2500 and 3000 µg/mL dihydrostreptomycin and at 3000 µg/mL of neomycin treatment. Only the highest concentration of dihydrostreptomycin (20000 µg/mL) reduced VERO cell viability significantly (P < 0.01). Based on or results we can assume the effect of different antibiotics in different concentrations on cell lines is various. Detection of antibiotic toxicity to animal cells is very important because of the increasing resistance of bacteria. One of the solutions is drug dose increasing, but only to a certain concentration, since the toxic effect over the therapeutic one will prevail, which we have also shown in this work.

The effect of different sample collection methods on rabbit blood parameters.

Nowadays great deal of research is physiological field is conducted on experimental animals and there is a lot of criticism from the wide public on methods used. Therefore, recently there is a lot of effort focused on the welfare of the animals. Main aim of this study is to determine the effect of experimental sample collection method on the selected parameters of stress. In the experiment two sample collections of rabbit blood from marginal ear vein were realized - first using standard method with one person fixing the animal and other collecting the blood using gently fixating the animal. In the second groups experimental method of inserting the experimental animal into a sack and further collection in dark was realized. During the experiment the levels of cortisol - main stress indicator in organism and other health parameters of animals including mineral profile and haematological parameters were observed. Our results show no significant changes in levels of cortisol but also a decreasing tendency in the sample from the second (dark) collection. Haematological parameters were generally in the reference values and any significant changes except levels of lymphocytes and percent of lymphocytes which shown significant increase in the second collection period were found. Also the levels of mean corpuscular haemoglobin and percent of neutrophils unveiled a significant decrease in values. Values of mineral profile parameters have indicated no significant changes except the levels of phosphorus. Based on the result we can state that the experimental sample collection has no effect on blood parameters of the animals but we spectated a statistically insignificant decrease in the levels of cortisol which can suggest that the dark collection is possibly less stressful to the animals.