Adriana Kolesárová

Environmental levels of cadmium, lead and mercury in brown hares and their relation to blood metabolic parameters.

The purpose of this study was to examine concentrations of selected heavy metals in the liver and kidney of brown hares (Lepus europaeus). In addition, correlations between heavy metals and biochemical parameters in blood plasma were determined. The average concentrations of heavy metals (mmol/L) ± SD were as follows: liver: Pb 0.221 ± 0.189, Cd 0.160 ± 0.140, Hg 0.021 ± 0.030, kidney: Pb 0.115 ± 0.125, Cd 1.570 ± 1.103, Hg 0.030 ± 0.053. The average concentrations of biochemical parameters in the blood plasma were as follows: Ca 3.16 mmol/L, P 2.19 mmol/L, Mg 1.40 mmol/L, Na 148.71 mmol/L, K 8.12 mmol/L, glucose 6.56 mmol/L, total proteins 56.49 g/L, urea 5.00 mmol/L, total lipids 1.40 g/L, bilirubin 3.97 μ mol/L, cholesterol 1.53 mmol/L, aspartate aminotransferase (AST) 6.06 μ kat/L and alanine aminotransferase (ALT) 1.94 μ kat/L. Average levels of hormones (ng/mL) were as follows: testosterone 2.94, androstendiol 0.13, estradiol 501.59, progesterone 6.63, oxytocin 328.60. Tissue analysis showed an accumulation of lead, cadmium and mercury in the liver and kidney of brown hares. There were no significant correlations between levels of heavy metals in liver, kidney, and biochemical parameters.

Effect of Lactobacillus fermentum and Enterococcus faecium strains on internal milieu, antioxidant status and body weight of broiler chickens.

The aim of the present study was to evaluate the functional efficiency of two probiotic strains Lactobacillus fermentum CCM 7158 and Enterococcus faecium M 74 given to the drinking water on internal milieu, antioxidant status and body weight of broiler chickens. The experiment was conducted on hybrid Hybro (n = 180). The feeding period lasted 42 days. Experimental chickens of E1 group received a probiotic preparation in drinking water with concentration of 1 × 10(9) colony forming units (CFU) of L. fermentum CCM 7158 in 1 g of nutrient medium and experimental chickens of E2 group concentration of 2 × 10(9) CFU of E. faecium M 74 in 1 g of nutrient medium. The control group of animals received water without any additives. Triglycerides content in serum mainly with L. fermentum strain against the control group was decreased. Calcium content in both experimental groups and significantly in E. faecium group was increased. Antioxidant status in both probiotic groups was significantly increased. The content of bilirubin in group with E. faecium M 74 was significantly increased. In conclusion, addition of a microbial feed additive (L. fermentum and E. faecium) increased serum calcium and iron level, decreased triglycerides content in blood and slightly increased body weight of broiler chickens.

In vitro study on the effects of lead and mercury on porcine ovarian granulosa cells

The heavy metals lead (Pb) and mercury (Hg) pose potential risks to sustainability of environment and thus to our future generations. General objective of this in vitro study was to examine the secretory activity of porcine ovarian granulosa cells after Pb and Hg administration and to outline the potential intracellular mediators of its effects. For this purpose, release of insulin-like growth factor I (IGF–I) and steroid hormone progesterone (P4), expression of proliferation- related (cyclin B1) and apoptosis-related (caspase-3) peptides was examined in porcine ovarian granulosa cells after heavy metals administration. Obtained data indicate Pb–induced inhibition of IGF–I release at lower doses (0.063 mg/mL and 0.046 mg/mL) by ovarian granulosa cells. However, P4 release was not influenced by Pb addition, while the expression of cyclin B1 and caspase-3 was induced by Pb addition. These results indicate that Pb can affect the pathway of proliferation and apoptosis of porcine ovarian granulosa cells through intracellular substances such as cyclin B1 and caspase–3. On the other hand, the P4 release by ovarian granulosa cells of pregnant gilts was stimulated by experimental Pb administration at doses of 0.25 mg/mL and 0.063 mg/mL and experimental Hg administration at doses 0.25 mg/mL and 0.083 mg/mL. P4 release by ovarian cells of pregnant gilts was not influenced by a combinatory dose of FSH (1.0 ng/mL) + Pb (0.083 mg/mL) + Hg (0.083 mg/mL) but it was inhibited by experimental administration of FSH (10 ng/mL) + Pb (0.25 ng/mL) + Hg (0.25 ng/mL). Possible involvement of heavy metals – Pb and Hg and pituitary hormone FSH, in the regulation of P4 release by porcine ovarian granulosa cells of pregnant gilts was noted. Data obtained from in vitro studies suggest the dose dependent association of heavy metals administration with the hormonal release by porcine ovarian granulosa cells. This association also depended on pregnancy of the gilts.

The effect of selected microbial strains on internal milieu of broiler chickens after peroral administration.

The aim of this work was to investigate the effect of probiotic preparation with multi-strains composition (Lactobacillus delbrueckii ssp. bulgaricus LAT 187, Lactobacillus acidophilus LAT 180, Lactobacillus helveticus LAT 179, Lactobacillus delbrueckii ssp. lactis LAT 182, Streptococcus thermophilus LAT 205 and Enterococcus faecium E 253) on internal milieu, antioxidant status and body weight of broiler chickens. Chickens were divided into three groups (C, control; E1 and E2, experimental groups). Experimental chickens received the probiotic preparation in drinking water. Probiotic preparation caused a significant increase of serum calcium and potassium content. In the group with higher dose of probiotic strains serum triglycerides level decreased. Total antioxidant status in groups with addition of probiotic strains showed higher values in comparison to control group. Serum albumin level was found to increase after consumption of probiotic preparation in group with higher dose of probiotic strains. Probiotic strains improved body weight in last observed weeks of feeding.

The effect of deoxynivalenol on the secretion activity, proliferation and apoptosis of porcine ovarian granulosa cells in vitro

The aim of this in vitro study was to examine the secretion activity, markers of proliferation and apoptosis in porcine ovarian granulosa cells (GCs) after deoxynivalenol (DON) addition. Ovarian granulosa cells were incubated with DON for 24h: 10, 100 and 1000 ng/mL, while the control group received no DON. The secretion of insulin-like growth factor I (IGF–I) and progesterone was determined by radioimmunoassay (RIA) and expression of cyclin B1, PCNA and caspase-3 by immunocytochemistry. IGF–I release by GCs was inhibited by DON, while progesterone release and the expression of cyclin B1 was stimulated by DON (at 1000 ng/mL but not at 10 and 100 ng/mL). PCNA expression was stimulated by DON (at 100 and 1000 ng/mL but not at 10 ng/mL). Caspase-3 expression was not influenced by DON treatment (at all doses). In conclusion, our results indicate, (1) a direct effect of DON on secretion of growth factor IGF-I and steroid hormone progesterone, (2) expression of markers of proliferation (cyclin B1 and PCNA) but not on the (3) expression of marker of apoptosis (caspase-3) in porcine ovarian granulosa cells. This in vitro study suggests the dose-dependent association of DON on porcine ovarian functions.

Resveratrol inhibits reproductive toxicity induced by deoxynivalenol.

The aim of this in vitro study was to examine the release of progesterone by porcine ovarian granulosa cells (GCs) after exposure to toxic concentrations of deoxynivalenol (DON), resveratrol (RSV), and their combination (DON with RSV). Ovarian granulosa cells were incubated without (control) or with treatments of natural substances at various doses for 24 h: RSV (10, 30 and 50 μg/mL) / DON (2000, 3000 and 5000 ng/mL), and their combination (10 μg/mL of RSV with 2000 ng/mL of DON; 30 μg/mL of RSV with 3000 ng/mL of DON; 50 μg/mL of RSV with 5000 ng/mL of DON). Progesterone was determined by radioimmunoassay (RIA). Progesterone release was significantly (P < 0.05) stimulated by RSV at the doses 50 μg/mL but not at 30 and 10 μg/mL and by DON treatment at all used doses (2000, 3000 and 5000 ng/mL). RSV in combination with DON stimulated significantly (P < 0.05) the progesterone release by GCs at the highest doses (50 μg/mL of RSV with 5000 ng/mL of DON). On the other hand, the stimulatory effect of RSV in combination with DON was significantly (P < 0.05) lower in comparison with alone DON effect. In conclusion, our results indicate, (1) the dose-depended stimulatory effects of RSV, DON and combination of RSV with DON on release of steroid hormone progesterone and (2) reduction of the stimulatory effect of DON by RSV. Our in vitro results suggest that reproductive toxicity of animals induced by a mycotoxin - deoxynivalenol can be inhibited by a protective natural substance – resveratrol.

Effects of dietary inclusion of Rhus coriaria on internal milieu of rabbits.

The aim of the present work was to investigate the effect of Rhus coriaria L. inclusion to the diet on some biochemical, haematological parameters and the level of antioxidant status of male rabbits. Adult rabbits were divided into five groups: one control (C) and four experimental groups. Experimental animals received sumac per os in feed in various doses (0.50%, 0.75%, 1.00% and 1.50%) for 90 days. Significant increase in PDWc (platelet distribution width) in E3 group when compared with control group was recorded. Sumac administration resulted in decreased cholesterol levels in all experimental groups vs. control group. Significantly lower level of cholesterol was found in E4 group with highest dose of sumac (1.50%). Higher values of total antioxidant status (TAS) and albumins were observed in all experimental groups in comparison with control group. A significant increase in TAS was detected in group E1 and E4. Concentrations of albumins were significantly higher in groups E3 and E4 vs. control group. Sumac administration had no significant effect on bilirubin content. In conclusion, these results show a positive effect of sumac consumption on antioxidant status and cholesterol level in adult male rabbits.

The effect of bee pollen on secretion activity, markers of proliferation and apoptosis of porcine ovarian granulosa cells in vitro.

The general objective of this in vitro study was to examine the effect of bee pollen on the release of insulin-like growth factor I (IGF–I) and steroid hormone progesterone, and expression of markers of proliferation (PCNA) and apoptosis (caspase-3) in porcine ovarian granulosa cells. Concentrations of IGF–I and progesterone were determined by RIA method and expression of PCNA and caspase-3 by immunocytochemistry. Bee pollen addition at the dose of 10 ng/mL significantly (P<0.05) inhibited IGF-I release by porcine ovarian granulosa cells. This growth factor was not influenced by 100 and 1000 ng/mL doses of bee pollen. Progesterone release by cells was not influenced by bee pollen addition at the doses of 10, 100 and 1000 ng/mL as used in our study. Similarly expression of PCNA and caspase-3 was not affected by bee pollen addition. The present study shows dose-dependent regulation of IGF–I by experimental bee pollen addition in vitro. Progesterone release, expression of PCNA and caspase-3 in porcine ovarian granulosa cells was not induced by pollen. Our results contribute to new insights regarding the possible effect of bee pollen on IGF-I release, which is important for regulation of porcine ovarian functions.

Cobalt-induced changes in the IGF-I and progesterone release, expression of proliferation- and apoptosis-related peptides in porcine ovarian granulosa cells in vitro

Cobalt (Co) is an essential element. The general objective of this in vitro study was to examine dose-dependent changes in the secretory activity of porcine ovarian granulosa cells after experimental Co administration and to outline the potential intracellular mediators of its effects. Concentrations of IGF-I and progesterone were determined by RIA and expression of cyclin B1 and caspase-3 by immunocytochemistry. IGF-I release by granulosa cells was stopped by Co addition at the concentration 1 mg/mL. Progesterone release by granulosa cells was decreased at the lowest Co addition (0.09 mg/mL). In our study the changes of the expression of proliferation related peptide cyclin B1 and apoptosis related peptide caspase-3 in ovarian granulosa cells was observed after experimental Co addition. The molecular pathways stimulated by Co through the expression of cyclin B1 and caspase-3 were found. In conclusion, this study provides novel evidence that cobalt is the factor which can initiate adverse effects in ovarian granulosa cells. These results contribute towards the understanding of mechanisms relating to endocrine disruptor-induced alterations in porcine ovarian granulosa cells.

Nickel-induced blood biochemistry alterations in hens after an experimental peroral administration.

The purpose of this study was to determine certain blood biochemical parameters in hens of Isa Brown breed (n = 20) after nickel administration. Animals were divided into four groups (K, P1, P2, P3). Experimental hens (n = 5; in each group) received nickel (NiCl2) as peroral administration in drinking water in various doses (P1 – 0.02 g NiCl2/L; P2 – 0.2 g NiCl2/L; P3 – 2.0 g NiCl2/L of drinking water) for 28 days. The last group – K (n = 5) was the control, receiving no nickel. Biochemical parameters of mineral profile (calcium; phosphorus; magnesium; sodium; potassium) and of energy and enzymatic profile [(glucose; total cholesterol; total proteins; triglycerides; alanine aminotransferase (ALT) aspartate aminotransferase (AST) gamma glutamyl transferase (GGT) and glutamatdehydrogenase (GLDH)] were analyzed in blood serum on Day 0, 7, 14, 21 and 28 of the experiment. Average levels of mineral metabolism parameters were relatively stable apart from calcium. The evaluation registered a significant decrease in calcium during the experiment mainly in the group with highest nickel concentration in drinking water. No significant differences were detected between groups in energy and enzymatic profile apart from the concentrations of ALT on Day 7. In conclusion, there were significant associations between nickel levels and calcium and ALT in blood serum of the hens. No significant differences were detected in other biochemical parameters of mineral profile (P, Mg, Na, K) and energy and enzymatic profile (glucose, total cholesterol, total proteins, triglycerides, AST, GGT and GLDH) after nickel administration. Our results may contribute to an evaluation of reference levels of analyzed parameters, to monitor the health and nutritional status of hens. In this study also the negative effect of nickel mainly on calcium metabolism was detected.