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Summary 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 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 micro mol/L, cholesterol 1.53 mmol/L, aspartate aminotransferase (AST) 6.06 micro kat/L and alanine aminotransferase (ALT) 1.94 micro 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.

Seasonal anoestrus in the ewe results from enhanced oestrogen negative feedback. Recent data have implicated the ventromedial preoptic area (vmPOA) as an important site of oestrogen action. This study addressed whether NO acts within the vmPOA to inhibit LH during seasonal anoestrus. In Experiment 1, microimplants containingNω-nitro-l-arginine methyl ester (l-NAME, NOS inhibitor),S-methyl thiocitrulline (SMTC, neural NOS (nNOS) inhibitor) or empty implants (control) were administered during mid-anoestrus to the vmPOA.l-NAME, but not SMTC, significantly increased LH pulse frequency. For Experiment 2, ewes in late anoestrus were administered 7-nitroindazole (7NI; nNOS inhibitor),l-NAME, SMTC, or empty implants. 7NI, but notl-NAME or SMTC, increased LH pulse frequency. In Experiment 3, the effects of microimplants and microinjections ofl-NAME were compared in mid-anoestrus. Microinjections ofl-NAME (300 nl at 10 μg/μl) increased LH pulse frequency, but microimplants did not. In late anoestrus, similar microinjections were ineffective. Taken together, the results of Experiments 1–3 suggested that NO inhibition may be stronger during the middle than at the end of seasonal anoestrus. To test this hypothesis, ewes in Experiment 4 received microinjection ofl-NAME or vehicle thrice during the non-breeding season; none of the treatments increased LH pulse frequency. These results indicate that NO plays a role in the vmPOA in suppressing LH secretion during seasonal anoestrus because NOS inhibitors were consistently stimulatory when LH pulse frequency was low. However, the inconsistent and modest effects of these inhibitors suggest that NO actions in this area cannot completely account for the effects of inhibitory photoperiod.

The ability of steroids to inhibit LH secretion is enhanced during undernutrition. To identify potential hypothalamic sites at which this enhancement may occur, we examined LH secretion in feed-restricted or fed young wethers treated with locally administered metabolites of testosterone. In experiment 1, microimplants containing crystalline estradiol-17beta (E) or cholesterol were administered via chronic guide tubes directed to the preoptic area (POA) or ventromedial hypothalamus (VMH) in fed or feed-restricted wethers. E treatment in the VMH decreased LH pulse frequency, pulse amplitude, and mean LH concentration in feed-restricted, but not fed, wethers. E may act in the POA to suppress LH under feed restriction, but definite conclusions cannot be drawn because of steroid-independent effects of feed restriction on LH pulse frequency. In experiment 2, the effect of dihydrotestosterone (DHT) in the VMH was determined. DHT administration to the VMH did not alter LH secretion in either feed-restricted or fed wethers. Thus the VMH is one site wherein E negative feedback is enhanced during feed restriction in the wether. In contrast, we found no evidence for enhanced responsiveness to androgen negative feedback within the VMH of feed-restricted wethers. We suggest that increased sensitivity within the VMH to E, but not to DHT, is important for suppressing LH secretion in undernourished male sheep.

Recent studies suggest that the endogenous opioid peptide, dynorphin, is an important mediator of progesterone negative feedback on GnRH pulse frequency in the ewe. These experiments tested this hypothesis by examining the effects of progesterone on dynorphin A concentrations in cerebrospinal fluid (CSF) collected from the third ventricle and expression of preprodynorphin (PPD) mRNA in hypothalamic nuclei. CSF was collected every 10 min for 5 h in three groups of ewes: 1) ovary-intact ewes during the luteal phase (d 6-7 of estrous cycle); 2) ewes 6-7 d after ovariectomy (OVX); and 3) OVX ewes treated for 6-7 d with implants that produced luteal-phase progesterone levels. Diencephalic tissue from these ewes was then collected and processed for in situ hybridization using an ovine cDNA probe against PPD. Progesterone treatment increased dynorphin A concentrations in CSF over that observed in untreated OVX ewes; CSF dynorphin A concentrations in ovary-intact ewes were midway between the other groups. OVX significantly decreased the number of PPD mRNA-expressing cells in the preoptic area (POA), anterior hypothalamic area (AHA), and arcuate nucleus (ARC), with no change seen in any other PPD-expressing nuclei. Progesterone treatment of OVX ewes restored PPD expression in the POA and AHA to levels seen in luteal-phase animals but had no effect on PPD expression in the ARC. These results are consistent with the hypothesis that progesterone acts via dynorphin neurons to inhibit pulsatile GnRH secretion and point to dynorphin neurons in the POA, AHA, and ARC as potential mediators of this action during the luteal phase.

Thyroid hormones are permissive for various species to enter seasonal anestrus. In the ewe they act centrally to permit the onset of potent estradiol-negative feedback responsible for anestrus, but the specific sites of action are unknown. Therefore, we tested whether T(4) replacement via chronic microimplants in any of five brain areas could reverse the reproductive effects of thyroidectomy. Diffusion of (125)I-T(4) from the microimplant was largely (98%) limited to a 1.2-mm radius. A marked decline in LH concentration in ovariectomized, estradiol-treated ewes was used as an index for anestrus. In experiment 1, all thyroidectomized (THX) ewes with microimplants in the medial preoptic area, A15 area, and medial basal hypothalamus failed to enter anestrus; instead, LH levels remained elevated, similar to those in untreated THX controls. In ventromedial preoptic area (vmPOA)-microimplanted ewes, only the two animals with the most caudal microimplants entered anestrus, as did thyroid-intact controls and THX ewes receiving icv or sc T(4) replacement. In experiment 2, all vmPOA-treated ewes with similar placements to those effective in experiment 1 along with all ewes microimplanted in the premammillary region entered neuroendocrine anestrus. Thus, the premammillary region and vmPOA are brain sites in which thyroid hormones act to permit the onset of seasonal anestrus.

Thyroid hormones permit the increase in response to estradiol negative feedback in ewes at the transition to anestrus. In this study, we tested whether the thyroid hormones are also required for steroid-independent seasonal changes in pulsatile LH secretion. In experiment 1, Suffolk ewes were ovariectomized and thyroidectomized (THX) or ovariectomized only (controls) in late November. LH pulse frequency and amplitude were measured for 4 h in December, April, May, June, and August. Pulse frequency was also measured in the presence of estradiol-containing implants during the breeding (December) and early anestrus (March) seasons. As expected, in the presence of estradiol, pulse frequency declined between December and March in control but not THX ewes. In the absence of estradiol, a seasonal decline in frequency and an increase in amplitude occurred in control ewes, concurrent with lengthening photoperiod. A similar trend was seen in THX ewes, but the seasonal changes were lower in magnitude and not significant. In experiment 2, the same protocol was used (pulse measurements in December, May, and June) with a larger THX group size (n = 7). Results were similar to those of experiment 1 for controls. In THX ewes, pulse frequency did not change over time and was significantly elevated relative to that of controls during the summer. Pulse amplitude in THX ewes tended to increase during summer and did not differ from pulse amplitudes in control ewes. These results demonstrate that thyroid hormones are required for steroid-independent cycles in LH pulse frequency; however, some seasonal changes in amplitude still occur in the absence of thyroid hormones. This finding contrasts with the changes in estradiol negative feedback at the transition to anestrus, which are entirely thyroid hormone dependent.

Oestradiol exerts a season-specific negative feedback effect on the GnRH/LH neurosecretory system of the Suffolk ewe. This neuroendocrine suppression is mediated in part by dopamine A15 neurones, but these neurones do not possess the oestrogen receptor. Based on indirect evidence, we hypothesized that oestrogen receptor-containing neurones in the ventromedial preoptic area (vmPOA) may be the initial step in a neuronal system whereby oestradiol suppresses GnRH secretion during the non-breeding season. To test this, three experiments were conducted using ovariectomized ewes receiving either empty or oestradiol-containing bilateral microimplants directed at the vmPOA or s.c. subcutaneous oestradiol-containing implants. In the first experiment, LH pulse frequency was measured on days 0, 1, 7 and 14 of treatment during seasonal anoestrus. In vmPOA oestradiol and s.c. oestradiol groups only, LH pulse frequency was suppressed on days 7 and 14, with maximal suppression evident by day 7. In the second experiment, this protocol was repeated during the breeding season, with LH pulses examined on days 0 and 7; LH pulse frequency did not change in any group. The third experiment tested if the effect of vmPOA oestradiol during anoestrus could be overcome by an injection of the dopamine-D2 receptor antagonist (-)-sulpiride. The vmPOA microimplants and s.c. oestradiol implants again suppressed LH pulse frequency and this was reversed by sulpiride in vmPOA oestradiol ewes. We conclude that oestradiol acts on cells in the vmPOA to stimulate a system involving dopamine neurones that inhibits GnRH/LH pulsatility in the anoestrous ewe.

This paper reports the results of accumulated selected metals concentrations (Fe, Mn, Zn, Cu, Ni, Co, Cr, Pb, Cd, Hg and meHg) in the muscle of five common Slovak fish species (Chub-Leuciscus cephalus, Common carp-Cyprinus carpio, Prussian carp-Carassius gibelio, Roach-Rutilus rutilus, and Wels catfish-Silurus glanis). Furthermore, correlations among the selected metals and order of metal accumulation in the fish muscle were determined. The concentrations of metals (mg/kg wet weight basis) ranged as follows: Fe 3.70-21.10; Mn 0.27-1.50; Zn 3.72-42.82; Cu 0.26-1.82; Ni 0.02-0.29; Co 0.06-0.28; Cr 0.09-0.28; Pb 0.08-34.59; Cd 0.06-2.76, Hg 0.34-3.64 and meHg 0.08-1.20. The level of lead and mercury exceeded the maximum allowed concentration in Slovakia by the Codex Alimentarius for safe human consumption (0.2 and 0.5 mg/kg, respectively) in the majority of samples (94.6 and 82.1%, respectively). Content of Cd (0.88 +/- 0.76 mg/kg wet weight) in the fish muscle exceeded maximum allowed levels (0.05 mg/kg) in all samples. An average, the order of metal concentrations in the fish muscle was: FeZnPbCdHgCuMnmeHgNiCrCo.

The purpose of this study was to assess concentration and correlation of selected heavy metals and level of some microbiological indicators (total bacteria count-TBC, mesophilic anaerobic sporulating bacteria-MASB) in the muscle of common carp (Cyprinus carpio). Fish were collected by seine net within the frame of pondfishing, in October (pond Horné Obdokovce--pond A) and December (pond Budmerice--pond B) 2003. Concentrations of selected metals were measured using an atomic absorption spectrophotometer Pye Unicam SP9. The concentrations of metals (mg/kg wet weight basis) ranged as follows: Fe 3.47-15.15; Mn 0.14-0.42; Zn 3.47-9.52; Cu 0.24-1.32; Co 0.05-0.17; Ni 0.07-0.42; Cr 0.08-0.19; Pb 0.11-0.30; and Cd 0.01-0.05. The count of MASB and TCB (in CFU/g) varied as follow: 1.12-7.76 x 10(3) and 0.03-7.59 x 10(6), respectively. Significant differences (P0.05) for Cu, Ni, and Cr bioaccumulation, as well as for count of MASB between ponds were recorded. An opposite trend of bioaccumulation was found in only 12 of 36 cases. Positive correlations between counts of MASB and accumulated heavy metals, except Ni and negative correlations between TBC and accumulated heavy metals, except Cr, were recorded, (P0.05) for TBC-Mn and TBC-Cd relationship. Lead concentrations exceeded the maximum values allowed concentration in Slovakia by Codex Alimentarius (0.2 mg/kg) by 60% for pond-A and 40% for pond-B. The level of MASB count exceeded the maximum allowed concentration in 100% by both ponds. On average, the order of metal concentrations in the fish muscle was: (Pond-A) FeZnCuMnPbCrNiCoCd; (Pond-B) FeZnCuMnPbNiCrCoCd.

The ability of steroids to inhibit LH secretion is enhanced during undernutrition. To identify potential hypothalamic sites at which this enhancement may occur, we examined LH secretion in feed-restricted or fed young wethers treated with locally administered metabolites of testosterone. In experiment 1, microimplants containing crystalline estradiol-17beta (E) or cholesterol were administered via chronic guide tubes directed to the preoptic area (POA) or ventromedial hypothalamus (VMH) in fed or feed-restricted wethers. E treatment in the VMH decreased LH pulse frequency, pulse amplitude, and mean LH concentration in feed-restricted, but not fed, wethers. E may act in the POA to suppress LH under feed restriction, but definite conclusions cannot be drawn because of steroid-independent effects of feed restriction on LH pulse frequency. In experiment 2, the effect of dihydrotestosterone (DHT) in the VMH was determined. DHT administration to the VMH did not alter LH secretion in either feed-restricted or fed wethers. Thus the VMH is one site wherein E negative feedback is enhanced during feed restriction in the wether. In contrast, we found no evidence for enhanced responsiveness to androgen negative feedback within the VMH of feed-restricted wethers. We suggest that increased sensitivity within the VMH to E, but not to DHT, is important for suppressing LH secretion in undernourished male sheep.

The purpose of this study was to assess concentration of selected metals (Fe, Mn, Zn, Cu, Ni, Co, Cr, Pb, Cd, and Hg) in the muscle of four common Slovak fish species (chub--Leuciscus cephalus, barbel--Barbus barbus, roach--Rutilus rutilus, and perch--Perca fluviatilis). Furthermore, correlations among the selected metals and order of metal accumulation in the fish muscle were determined. An electrofishing technique was used for collecting the fish from the Nitra River (Slovakia) in September 2003. Concentrations of selected metals were measured using an atomic absorption spectrophotometer Pye Unicam SP9. The concentrations of metals (mg/kg wet weight basis) ranged as follows: Fe 3.41-15.14; Mn 0.20-0.81; Zn 3.51-15.64; Cu 0.25-0.78; Ni 0.07-0.25; Co 0.05-0.19; Cr 0.11-0.42; Pb 0.20-5.81; Cd 0.06-0.56, and Hg 1.35-6.52. Significant correlations (P0.05) between Fe-Cu, Fe-Ni, Fe-Cr, Mn-Ni, Mn-Cr, Mn-Cr, Cu-Ni, and Ni-Cr were observed. The level of lead exceeded the maximum allowable concentration in Slovakia by Codex Alimentarius for safe human consumption (0.2 mg/kg) in the majority of samples (97.2%). Content of Cd (0.23 +/- 0.13 mg/kg wet weight) and Hg (2.85 +/- 1.22 mg/kg wet weight) in the fish muscle exceeded maximum allowed levels in all samples. On average, the order of metal concentrations in the fish muscle was: FeZnHgPbCuMnCdCrNiCo.

Cadmium accumulation in ovaries and uterus after i.p. and p.o. administration are reported. Time and dose dependent increase in cadmium concentration was found. More than 10 times higher accumulation is after i.p. administration in comparison with p.o. administration in ovaries as well as in uterus. There is very high correlation relationship among growing follicles and stroma in general as well as in all groups. So, we can report that with the increasing number of growing follicles there is lower amount of stroma. Generally we have found mean correlation relationship among cadmium content in ovaries and the percentual amount of growing follicles and stroma. We can say that with higher cadmium content in ovaries there is lower amount of primary follicles and stroma, but more growing follicles. On the other hand our previous study have described higher incidence of follicular atresia, which affect mainly growing follicles. In evaluation of numer of ovarian follicles to cadmium content, we report t...