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In many European countries over the last few decades, arable fields dominate agricultural landscapes, leading to very intensive land-use practices. This seems to be the main cause of population declines for numerous farmland species, including the European brown hare (Lepus europaeus Pallas, 1778). The Research Institute for Animal Production (National Agricultural and Food Centre—NPPC, Luzianky, Slovakia) has been engaged in a long monitoring project (a project currently running), collecting certain indicators of brown hare population dynamics during hunting season from 1987 to 2023 in the Slovak Danubian Lowland. In the same macro-area (Čiližská Radvaň and Lehnice), a study was conducted on the influence of permanent semi-natural vegetation in relation to recruitment, population density and production. The entire monitored period was aggregated into 5-year intervals (for a total of seven time intervals), with the aim of analyzing the brown hare population dynamics. Spring hare density in the Danubian Lowland is currently 20.8 hares/km2, with harvests of 4.6 hares/km2. During the monitoring period, bag animals have been examined following the regular hunting operations for the purpose of age determination (weight of eye lenses), sex ratio and productivity. There was a large positive effect of set-aside with special mixtures created for hares in large-scale farmed agrarian landscapes on brown hare density, bag and recruitment. In-model hunting grounds with such set-asides increased the spring stock by 25%, bag by 100% and recruitment by 20%. This study reveals that the management of European brown hare is not sustainable in the Slovak Danubian Lowland, and the population is decreasing. This is proven through the decline in harvest brown hares and by population dynamic parameters. Our data suggest that improvements in the habitat quality of arable landscapes by the adoption of permanent semi-natural vegetation may be more effective in the increase in the brown hare population. [ABSTRACT FROM AUTHOR]

The goal of this study was to monitor the accumulation of aflatoxin B1in the liver and kidney of brown hares (Lepus europaeusPall) in the region of south-western Slovakia. A total of 65 samples were involved for analysis by RIA method. Brown hares were divided into the groups according to age, sex and season (month). The sex was determined visually after shooting, and the age was assigned from dried eye lens. The average concentration of AFB1in the liver of hares was 0.54 ± 0.053 µg/kg, and lower values were measured in the kidney (0.41 ± 0.038 µg/kg). The significantly (P< 0.05) higher values were found in winter months when compared to summer months. According to the age, juvenile animals showed significant higher accumulation of B1in both organs than adults (P< 0.05). Wild animals can serve as a good model of real environmental contamination. Thus, monitoring of risk factors such as mycotoxins in the environment is important with regard to public health, as game animals constitute an important part of food chain for humans. [ABSTRACT FROM PUBLISHER]

The objective of thisin vitrostudy was to examine dose–dependent changes in the secretion activity [progesterone (P4) and insulin-like growth factor–I (IGF–I)] of porcine ovarian granulosa cells after experimental mercury (Hg) administration, including its apoptotic potential so as to ascertain the possible involvement of Hg in steroidogenesis. Ovarian granulosa cells were incubated with mercuric chloride [mercury (II) chloride or HgCl2] at the doses 50–250 μg mL−1for 18 h and compared with control group without Hg addition. Release of P4and IGF–I by ovarian granulosa cells was assessed by RIA and apoptosis by TUNEL assay. Observations show that P4release by granulosa cells was significantly (P< 0.05) inhibited at all the doses, while IGF-I release was not affected at any of the doses used, although a decreasing trend in the release of IGF–I was noted in comparison to control. An increasing trend of apoptosis of granulosa cells was noted, the difference being significant (P< 0.05) only at the dose 130 μg mL−1HgCl2, in comparison to control. Obtained data suggest a direct effect of Hg on the release of steroid hormone progesterone but not growth factor IGF-I, and a dose-dependent effect on apoptosis of porcine ovarian granulosa cells. Results indicate the interference of Hg in the pathways of steroidogenesis and apoptosis of porcine ovarian granulosa cells. [ABSTRACT FROM AUTHOR]

This study aimed at obtaining the data on the occurrence, levels and correlations of organic pollutants present in game animals (n = 75, Brown hare, Lepus europaeus Pall.) in the region of south-western Slovakia. The analyses performed included dichlorodiphenyltrichloroethane (DDT), hexachlorobenzen (HCB), alpha- and beta hexachlorocyclohexane (α+β-HCH), gamma-hexachlorocyclohexane (γ-HCH), and polychlorinated biphenyls (PCB-delor, commercial mixture of PCB congeners). A gas chromatograph with an electron capture detector was used for the analysis. PCB-delor and DDT were accumulated significantly in the highest level (0.105±0.059 mg/kg; 0.070 mg/kg) in depot fat in brown hares; however maximum permissible limits for the observed pollutants were not exceeded. Significantly higher concentrations of DDT, HCB, γ-HCH, and PCB-delor were found in adult animals when compared with juvenile hares. Gender and season had no effect on the accumulation of observed pollutants. Moderately positive correlation was found between PCB-delor and DDT (r = 0.59). Monitoring of environmental pollution with polychlorinated hydrocarbons is important with regard to public health, as game animals constitute an important part of food chain also for humans. [ABSTRACT FROM PUBLISHER]

The aim of this study was to monitor accumulation of cadmium (Cd), mercury (Hg), zinc (Zn), copper (Cu) and cobalt (Co) in the muscle, liver and kidney of wild boar (Sus scrofa scrofa) from hunting place of western Slovakia and the correlations among the observed elements. A total of 120 samples were involved for analyses by atomic absorption spectrophotometry (AAS). The significantly highest accumulation of Cd in the kidney followed by the liver and muscles was found. Zn accumulated mainly in the liver. Significantly lower values were found in the kidney followed by the muscle. The concentration of Cu was significantly lowest in the muscle when compared to the liver and kidney. Hg and Co accumulated mainly in the kidney, followed by the liver and muscle of wild boars, but without significant differences. In the muscle of wild boar moderately positive correlation between Zn and Cu (r = 0.59), Cd and Co (r = 0.51), Cu and Co (r = 0.33), and Zn and Hg (r = 0.36) were found. In the liver moderately positive correlation between Cd and Hg (r = 0.39) was detected. Moderately positive correlation between Zn and Cu (r = 0.40) was noted for the kidney. [ABSTRACT FROM PUBLISHER]

The aim of our work was to analyse the expression pattern of embryonic stem cell specific markers in rabbit embryos and epiblast cells. There are well-known genes that direct pluripotency in the mouse, such as Oct4, Nanog, but their expression pattern during the rabbit embryonic development is not known yet. Using culture medium supplemented with two inhibitors [(2i: PD032590-(PD), CHIR99021-(CH); embryonic stem cell (ESC) lines could be derived efficiently from mouse and rat embryos. We cultured the rabbit embryos from eight-cell-stage in RDH, RDH+2i culture medium until the blastocyst stage. The expression level of epiblast specific Oct4 and Nanog, trophoblast specific Cdx2 and hypoblast specific Gata4 and Gata6 were examined in blastocysts and attached embryoblasts. The expression level of Oct4 and Nanog was significantly higher in blastocysts, cultured in RDH+2i culture medium, but there was no significant difference in CDX2, GATA4, GATA6 expression on mRNA and protein level, compared to embryos cultured in RDH medium. We hope that we can maintain the pluripotency of the epiblast derived stem cells in 2i containing medium during long term cultivation. [ABSTRACT FROM AUTHOR]

The objectives of the present study were to: (i) examine the in vitro dose response of rabbit spermatozoa motility to the antifertility agent gossypol (GOS) and (ii) determine whether filtered (FIL) and unfiltered (UNFIL) GOS differ in their magnitude of effect. Rabbit semen belonging to adult males (n = 5; 12-14 months) were cultured with UNFIL GOS and FIL GOS (5% solution) and subsequently diluted (1:1-7) for analysis using a Computer Assisted Semen Analyzer (CASA) system in 5 time periods (0, 60, 120, 180 and 360 minutes). At Time 0, no significant change in rabbit spermatozoa motility (MOT) and progressive motility (PROG) with GOS FIL was noted, while increases were observed with GOS UNFIL. At Time 60, weak changes were noted for MOT and PROG. After 120 minutes of culture with both GOS FIL and GOS UNFIL, MOT and PROG decreased significantly in some experimental groups. However, no differences were recorded for both the parameters at Times 180 and 360, with the exception of PROG in the GOS UNFIL category (groups A, B, E, F and G), where a significant decrease was noticed. Detailed evaluation of the distance and velocity parameters revealed reduction in all these studied markers after 60 and 120 minutes of in vitro culture with both GOS FIL and GOS UNFIL, indirectly confirming the PROG decrease. Straightness (STR), linearity (LIN), wobble (WOB), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) mostly remained unaltered at all time periods for GOS FIL, where as some minor alterations were noticed in GOS UNFIL category for STR, LIN, WOB, ALH and BCF parameters at Time 0, 60 and 120. The present study confirms the dose and time dependent alterations of rabbit spermatozoa motility parameters by GOS. The GOS dynamics in our experiment shows that rabbit spermatozoa as a biological material can indicate a GOS inhibition of motility. Obtained data for the first time indicates a higher immobilizing potential of unfiltered GOS in comparison to filtered GOS in its inhibitory action of spermatozoa motility parameters in rabbits. [ABSTRACT FROM AUTHOR]

Cytotrophoblast (CTB) of the early gestation human placenta are bipotent progenitor epithelial cells, which can differentiate into invasive extravillous trophoblast (EVT) and multinucleated syncytiotrophoblast (STB). Trophoblast stem cells (TSC), derived from early first trimester placentae, have also been shown to be bipotential. In this study, we set out to probe the transcriptional diversity of first trimester CTB and compare TSC to various subgroups of CTB. We performed single-cell RNA sequencing on six normal placentae, four from early (6-8 weeks) and two from late (12-14 weeks) first trimester, of which two of the early first trimester cases were separated into basal (maternal) and chorionic (fetal) fractions prior to sequencing. We also sequenced three TSC lines, derived from 6-8 week placentae, to evaluate similarities and differences between primary CTB and TSC. CTB clusters displayed notable distinctions based on gestational age, with early first trimester placentae showing enrichment for specific CTB subtypes, further influenced by origin from the basal or chorionic plate. Differential expression analysis of CTB from basal versus chorionic plate highlighted pathways associated with proliferation, unfolded protein response, and oxidative phosphorylation. We identified trophoblast states representing initial progenitor CTB, precursor STB, precursor and mature EVT, and multiple CTB subtypes. CTB progenitors were enriched in early first trimester placentae, with basal plate cells biased toward EVT, and chorionic plate cells toward STB, precursors. Clustering and trajectory inference analysis indicated that TSC were most like EVT precursor cells, with only a small percentage of TSC on the pre-STB differentiation trajectory. This was confirmed by flow cytometric analysis of 6 different TSC lines, which showed uniform expression of proximal column markers ITGA2 and ITGA5. Additionally, we found that ITGA5 + CTB could be plated in 2D, forming only EVT upon spontaneous differentiation, but failed to form self-renewing organoids; conversely, ITGA5-CTB could not be plated in 2D, but readily formed organoids. Our findings suggest that distinct CTB states exist in different regions of the placenta as early as six weeks gestation and that current TSC lines most closely resemble ITGA5 + CTB, biased toward the EVT lineage., Competing Interests: Declaration of interests Authors declare that they have no competing interests.

Investigations regarding European brown hare syndrome virus (EBHSV) in European brown hares (Lepus europaeus) in Slovakia were undertaken in order to detect the possible presence of EBHSV and to evaluate its phylogenetic position. Liver and/or serum samples were obtained from 135 European brown hares shot by hunters in eight regional hunting areas. From 36 animals corresponding liver and serum samples were available; from the remaining 49 and 50 animals only liver or serum samples were examined, respectively. Samples were tested for antibodies against EBHSV and for viral RNA by reverse transcriptase-polymerase chain reaction (RT-PCR) and RT-PCR products were subsequently sequenced. Additionally, matrilinear hare haplotypes were analyzed in order to detect potential familial susceptibility to EBHSV. Sixty-three of 86 sera contained antibodies against EBHSV, whereas 15 of 85 liver samples were PCR positive. Of the latter, 14 were sequenced, revealing three new strains of EBHSV. Fifteen different matrilinear haplotypes were identified, but no correlation was found between haplotype and susceptibility to EBHSV infection. Our findings confirmed the existence of EBHSV in Slovakia and reinforce the need for determining EBHSV status when importing hares for restocking.