Volume 67, Issue 4

Heme oxygenase-1 (HO-1) is the rate limiting enzyme in heme catabolism and degrades heme to carbon monoxide, biliverdin, and ferrous iron. HO-1 transcriptional induction occurs in response to multiple forms of chemical, physical stress and cytokines. HO-1 confers cytoprotection by inhibiting apoptosis, oxidative stress, and inflammation. Hepatitis C virus (HCV) is a major cause of cirrhosis and hepatocellular carcinoma. It has been shown that HO-1 induction and HO-1 products interfere with replication of HCV and markedly decreased HCV replication. However, a growing body of evidence indicates that induction of HO-1 may be involved in carcinogenesis and can play a role in the metastasis and growth of tumors. The antioxidant, antiviral activity of HO-1 makes it the cytoprotective enzyme for liver tissue in HCV infection, and induction of HO-1 can be suggested as a future therapeutic approach. However, the role of HO-1 in tumor growth should not be ignored.

An extracellular alkaline protease-producing Vibrio sp. was isolated from mangrove sediments of Vellar estuary. A 9.36-fold purification was achieved by a three-step purification procedure and the molecular weight of the enzyme was determined as 33 kDa by SDS-PAGE. The enzyme was active in a broad range of pH (6.0–11.0) and temperature (30–70°C), the optimum being at pH 9.0 and temperature 55°C. The enzyme was stable at alkaline pH range of 9–11 and up to a temperature of 60°C, after incubation for 1 h. Metals like Co2+, Hg2+, Ni2+ and Cu2+ inhibited the enzyme activity, whereas Fe2+, Ca2+ and Mn2+ were found to enhance the activity. The protease was found to be highly stable in the presence of oxidizing agents like H2O2, detergents such as SDS and Triton-X-100 and also some of the commonly used commercial detergents. The organic solvents like xylene, isopropanol, hexane and benzene were found to enhance as well as stabilize the enzyme activity. The extracellular production of the enzyme, the pH and thermal stability, and the stability in presence of oxidants, surfactants, commercial detergents and organic solvents, altogether suggest that it can be used as a laundry additive.

The cyclodextrin glucanotransferase from Paenibacillus pabuli US132 (US132 CGTase) was engineered using a rational approach in an attempt to provide it with anti-staling properties comparable to those of the commercial maltogenic amylase (Novamyl). The study aimed to concurrently decrease the cyclization activity and increase the hydrolytic activity of US132 CGTase. A five-residue loop (PAGFS) was inserted, alone or with the substitution of essential residues for cyclization (G180, L194 and Y195), mimicking the case of Novamyl. The findings indicate that, unlike the case of the CGTase of Thermoanerobacterium thermosulfurigenes strain EM1 whose initial high hydrolytic activity was exceptional, these mutations completely abolished the cyclization and hydrolytic activities of the US132 CGTase. This suggests that those mutations are not able to convert conventional CGTases, whose hydrolytic activities are very weak, into hydrolases. Accordingly, and for the first time, a structural barrier at subsite −3 was advanced as an influential factor which might explain the low hydrolytic activity of conventional CGTases.

We made an attempt to isolate and purify metagenomic DNA from chitin enriched soil. In this communication we report a modified direct lysis method for soil DNA extraction including initial pre-lysis washing of sample, followed by a rapid polyvinylpyrrolidone-agarose-based purification and electroelution of DNA using Gene-capsule™ assembly. Rapidity was achieved using low molarity conducting media (sodium-borate buffer) for electrophoresis by reducing run time for both the gel electrophoresis and electroelution. Extracted DNA was sufficiently pure and of high quality, evidenced by amplification of 16S rDNA and chitinase genes by PCR. Metagenomic nature of the DNA was confirmed by running V3 (16S rDNA) region amplicons using denaturing gradient gel electrophoresis. This method requires 30 min for purification, and less than 2 h for complete execution of protocol and becomes the first report on the isolation of metagenomic DNA from soil naturally enriched for chitin.

Two chimeric genes, XynA-Bs-Glu-1 and XynA-Bs-Glu-2, encoding Aspergillus sulphureus β-xylanase (XynA, 26 kDa) and Bacillus subtilis β-1,3-1,4-glucanase (Bs-Glu, 30 kDa), were constructed via in-fusion by different linkers and expressed successfully in Pichia pastoris. The fusion protein (50 kDa) exhibited both β-xylanase and β-1,3-1,4-glucanase activities. Compared with parental enzymes, the moiety activities were decreased in fermentation supernatants. Parental XynA and Bs-Glu were superior to corresponding moieties in each fusion enzymes because of lower Kn higher kcat. Despite some variations, common optima were generally 50°C and pH 3.4 for the XynA moiety and parent, and 40°C and pH 6.4 for the Bs-Glu counterparts. Thus, the fusion enzyme XynA-Bs-Glu-1 and XynA-Bs-Glu-2 were bifunctional.

Recent field research on grassland fungi in Slovakia resulted in collection of rare geoglossaceous fungi. Previously published data on the distribution of the genus Geoglossum in Slovakia have been extended: G. glutinosum (11 sites), G. glabrum (3 sites), G. cookeanum (1 site), G. umbratile (9 sites) and G. fallax (3 sites). The last three species are new for the territory. Each treated species is described and ecological preferences and threat categories are discussed. A field identification key of accepted species is included.

The authors bring actual state of recent occurrence of the rarest willows species Salix myrtilloides, its critically endangered population from northern part of the Western Carpathians. First original data on recent existence and distribution of the interspecific hybrid Salix × onusta (= S. myrtilloides × S. aurita) completes at the same time knowledge on hitherto unknown past occurrence of S. myrtilloides in the Western Carpathians and extinction of that population.

In this study, the anatomical features of the leaf and stem, besides the nutlet characteristics of some Teucrium sect. Isotriodon (Lamiaceae) taxa in Turkey, T. montbretii Betham subsp. montbretii, T. montbretii subsp. pamphylicum P. H. Davis, T. odontites Boiss. & Bal., T. cavernarum P. H. Davis, T. antitauricum T. Ekim, along with an isolated population of T. montbretii (T. montbretii subsp.) were investigated. The anatomical studies revealed that the taxa share generally similar anatomical characters, such as thicker upper leaf cuticles and larger upper leaf epidermal cells compared to lower ones and diacytic to anomocytic stomata on the leaves. However, the portion of the mesophyll occupied by palisade parenchyma and the occurrence of mucilage cells in leaf epidermis shows difference among the taxa. Furthermore, the studied taxa have general stem characteristics of the Lamiaceae family, except for having poorly developed collenchyma at the corners. With the amphistomatic leaves and developed sclerenchymatic tissue in the leaf median vein, T. cavernarum is seperated from the other taxa. Trichome types on the vegetative organs and nutlet shape and sculpturing are generally the same or similar in the studied taxa, but trichomes on the nutlets are different among them. Based on nutlet characteristics and some morphological ones, it was revealed that the isolated population of T. montbretii represent a new subspecies, T. monbretii subsp. yildirimlii M.Dinç & S.Doğu subsp. nov.

Enhanced nitrogen (N) levels accelerate expansion of Calamagrostis epigejos and Arrhenatherum elatius, highly aggressive expanders displacing original dry acidophilous grassland vegetation in the Podyjí National Park (Czech Republic). We compared the capability of Calamagrostis and Arrhenatherum under control and N enhanced treatments to (i) accumulate N and phosphorus (P) in plant tissues, (ii) remove N and P from above-ground biomass during senescence and (iii) release N and P from plant material during decomposition of fresh formed litter. In control treatment, significantly higher amounts of total biomass and fresh aboveground litter were observed in Calamagrostis than in Arrhenatherum. Contrariwise, nutrient concentrations were significantly higher (11.6–14.3 mg N g−1 and 2.3 mg P g−1) in Arrhenatherum peak aboveground biomass than in Calamagrostis (8.4–10.3 mg N g−1 and 1.6–1.7 mg P g−1). Substantial differences between species were found in resorption of nutrients, mainly P, at the ends of growing seasons. While P concentrations in Arrhenatherum fresh litter were twice and three times higher (1.6–2.5 mg P g−1) than in Calamagrostis (0.7–0.8 mg P g−1), N concentrations were nearly doubled in Arrhenatherum (13.1–15.6 mg N g−1) in comparison with Calamagrostis (7.4–8.7 mg N g−1). Thus, the nutrients (N and mainly P) were retranslocated from the aboveground biomass of Calamagrostis probably more effectively in comparison with Arrhenatherum at the end of the growing season. On the other hand, Arrhenatherum litter was decomposed faster and consequently nutrient release (mainly N and P) was higher in comparison with Calamagrostis which pointed to different growth and nutrient use strategies of studied grass species.

The gibberellin 20-oxidase gene (GA20) plays an important role in plant growth and development. Differential expression of Zoysiagrass (Zoysia japonica Steud.) gibberellin 20 oxidase gene (ZjGA20) induced by abiotic stresses has not been reported. In this investigation, we first reported the differential expression of ZjGA20 in different Z. japonica tissues including root, young leaf, senescent leaf, blade, sheath, and stolon, as well as differential expression induced by abiotic stresses including low temperature (4°C), H2O2(8 μM), salt stress (250 mM NaCl), 25% PEG6000, and high temperature (42°C) by Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). Higher expression of ZjGA20 was observed in young leaf and sheath, compared to root, senescent leaf, blade, and stolon. Among different abiotic stresses, expression of ZjGA20 decreased under low temperature, 25% PEG6000, and high temperature. The highest expression of ZjGA20 was obtained when plants were treated with 8 μM H2O2 for 10 h and with 250 mM NaCl for 5 h. The analysis of the MDA content, POD activity, and permeability of the plasma membrane demonstrated that application of exogenous GA3 recovered tissue damage derived from low temperature treatment. In addition, the expression of ZjGA20 increased under low temperature stress. These results demonstrated that expression of ZjGA20 was regulated by abiotic stresses and the damage derived from abiotic stresses could be rescued by exogenously applied plant growth regulator GA3. Further more, exogenous gibberellin and salicylic acid (SA) alleviated the growth inhibition and death of the seedlings under stresses. The SA content in the seedlings treated with 80 μM GA3 was far greater than the control (with H2O) and plants under stress treatments. These data suggest that exogenous addition of GA3 is able to counteract the inhibitory effects of these adverse environmental conditions in Zoysiagrass growth through modulation of SA biosynthesis. This is the first study of differential expression of gibberellin 20 oxidase gene and growth regulation of GA3 in Zoysiagrass under stresses.

Prosopis strombulifera, a common legume in high-salinity soils of Argentina, is a useful model for elucidation of salt tolerance mechanisms and specific biochemical pathways in halophytes, since its NaCl tolerance exceeds the limit described for most halophytic plants. We analyzed the effects of the increasing concentration of two main soil salts, Na2SO4 and NaCl, on growth parameters of P. strombulifera, chlorophyll levels, and content of jasmonates (JAs) and polyamines (PAs), which are key molecules involved in stress responses. P. strombulifera showed a halophytic response (growth promotion) to NaCl, but strong growth inhibition by iso-osmotic solutions of Na2SO4. Chlorophyll levels, number of leaves and leaf area were also differentially affected. An important finding was the partial alleviation of SO42− toxicity by treatment with two-salt mixture. JAs are not directly involved in salt tolerance in this species since its levels decrease under all salt treatments. Beneficial effects of Putrescine (Put) accumulation in NaCl treated plants maybe inferred probably associated with the antioxidative defense system. Another novel finding is the accumulation of the uncommon PA cadaverine in roots under high Na2SO4, which may be related to SO42− toxicity.

Callose plays important roles in a variety of processes of plant development, and/or in a response to a range of biotic and abiotic stresses. In the current work we have studied and compared the effect of lead, cadmium and arsenic on accumulation of newly formed callose deposits in the roots of maize and soybean. We observed formation of characteristic callose deposits in the root cell walls, probably associated with plasmodesmata, depending on the type of metal and the plant species investigated. Further, the callose turnover was analysed by measuring of total callose content as well as activities of total β-(1,3)-glucanases in roots. The latter enzymes are responsible for callose depletion, and their possible role during metal stress has previously been proposed. However, neither of these biochemical values appeared to be sufficiently reliable for scoring the altered callose turnover (including local deposits) in plant tissue. The microscopical observations are discussed in light of the biochemical data obtained.

The aim of this study was to determine the effect of exogenously applied Si on the growth and physiological parameters of sorghum [Sorghum bicolor (L.) Moench] cultivated in hydroponics with elevated zinc concentrations (75 μM and 150 μM Zn). Increased concentrations of Zn inhibited root growth and biomass production of roots and shoots. Application of Si individually showed a positive effect on root growth but negatively affected production of fresh and dry biomass of roots and shoots. On the other hand, silicon in combination with Zn significantly reduced the inhibitory effect of Zn on root growth but did not positively affect biomass production of roots and shoots. Accumulation of Zn in plant tissues increased with increasing Zn concentration in nutrient solution, but application of Si in combination with Zn did not significantly influence Zn accumulation in roots. Completely opposite results were found in Si accumulation in roots treated with Si in combination with Zn. Interaction of these ions resulted in considerable increase of Si accumulation in roots which almost doubled in comparison to individal Si treatment. Impact of Zn on the activity of some antioxidant enzymes was equivocal and differences were observed also between two Zn concentrations. Individual application of Si resulted in significant increase in the activity of all studied antioxidant enzymes but Si in combination with Zn mostly negatively affected their activity except the activity of catalase (CAT) which was the highest in roots grown in solution containing both Si and Zn ions. Comparing all obtained data we can assume that Si applied in combination with Zn did not significantly alleviate Zn toxicity in young sorghum except the growth of primary seminal root and further experiments are required for better understanding of their interaction.

Limited availability of phosphate ion (Pi) reduces plant growth in natural ecosystems. Here, we report the functional effects of overexpressing an Arabidopsis thaliana purple acid phosphatase encoding gene, AtPAP18, in Nicotiana tabbacum as a crop model plant. Transgenic tobacco plants exhibited significant increases in acid phosphatase activity, total P and Pi contents leading to improved biomass production in both Pi-deficient and Pi-sufficient conditions. Transient expression of AtPAP18::green fluorescent fusion protein in onion epidermal cells indicated that AtPAP18 is a dual-targeted protein, which is detected mainly in the apoplast of the cells after 24 h and in the vacuole after 72 h. Possibly, AtPAP18 protein confers efficient retrieval of Pi from bonded extracellular compounds as well as expendable intracellular Pi-monoesters and anhydrides. These data clearly indicate that overexpression of AtPAP18 gene offers an effective approach for reducing the consumption of chemical Pi fertilizer through increased acquisition of soil Pi and mobilization of internal resources.

Leech taxonomy is based on unstable morphological characters. The overall level of genetic variability and species differentiation is unknown. Using the RAPD assays genetic diversities and genetic similarities were estimated in twelve species collected in North-Eastern Poland and representing three families and five subfamilies. Ten primers revealed 204 reproducible bands. Genetic diversities varied from 0.099 to 0.219 classifying studied species among variable invertebrates. Total 45 markers comprised 22% of all amplified bands were unique for species thus enabling their identification. Genetic similarities among species (0.528–0.811) evidenced several stages of differentiation, which is mirrored in the current taxonomy. The UPGMA and multidimensional scaling (nMDS) based on our RAPDs are congruent and reflected traditional division into “Rhynchobdellida” and Arhynchobdellida. The RAPD approach proved to be an effective tool in population and evolutionary studies of leeches. For the first time, genetic parameters were estimated enabling to compare leeches with outcomes from other animals.

Emys orbicularis is receding from Europe, mainly due to anthropogenic habitat changes. Its parasite, Placobdella costata, is widely distributed within both the former and the present distribution range of the host. Though closely associated with the mud turtle, it may have other hosts (birds, amphibians, reptiles). Its reproductive period coincides with that of its host’s migration to the breeding grounds, thus facilitating dispersal of the parasite. Based on literature data we have analyzed the geographic spread of P. costata and mud turtle to observe the possible overlap of their habitats. Observations on the population of mud turtle and the associated leech species (P. costata) were carried out in eastern Poland — Podlasie Lowland. The studies were conducted in spring and summer in 1986–1993. The leeches were collected from the turtles caught in the water and on land. Observations showed that most leeches were found on turtles inhabiting the lake or moving to a breeding area. The greatest intensity of invasion was observed in June and July and that most leeches were observed in female E. orbicularis characterized by greater length of the carapace and weight, compared with males and juvenile individuals.

The aim of this study was to reveal the spatial and temporal variation in biomass, density, and size structure of the invasive clam, Corbicula fluminea in the Danube River catchment and to explore the environmental factors determining these patterns. Samples were taken seasonally during the years 2007 and 2008 at 15 sites located along a river continuum from a second order stream to the Danube River. C. fluminea was the most abundant species in the main arm of the Danube; however, it was found in great density in the side arms, too. In the Ipoly River it only occured close to the conjuction, and it was not present at all in the second and third order streams. Regression models were developed to predict dry mass (DW, g) from linear body dimension [shell length (L), mm]. For the description of length-dry mass relationship, the power function (DW = a * Lb) was applied. The highest density, biomass, and mean shell length were 178 ind. m−2, 24.9697 g DW m−2, and 21.99 mm, respectively. ANOVA detected significant spatial variation in biomass; however, significant temporal variation was not recorded between the years, only among the seasons. Multiple linear regressions were used to find the relationship between the biomass and the environmental parameters. The coarse and very fine sediment fractions and the high water temperature provided suitable habitat for C. fluminea. Although statistically significant correlation was not detected between the chlorophyll-a content of water and the biomass, the highest biomass values were recorded at sites with high chlorophyll-a value.

A study on the relative growth and reproductive cycle of the purple dye murex (Bolinus brandaris) was performed from March 2007 to February 2008 in two populations with contrasting imposex levels (Carthage Byrsa — CB in the small Gulf of Tunis and Menzel Abderrahmane — MA in the Bizerta Lagoon). Both populations presented balanced sex ratios. Comparison of allometric relationships established between linear and ponderal variables (sexes confounded) revealed higher growth in CB than in MA. Mature individuals were found throughout the year, except for September in MA. Spawning periods occurred from March–April to May and from June to August in both sites. Spawning was associated to an increase in seawater temperature. Besides increasing the current knowledge on the biology of B. brandaris from the southern Mediterranean, the information gathered in the present study constitutes a useful baseline for the sustainable management of local wild stocks, namely by prohibiting collection of snails during the spawning season.

Cholinesterase (ChE) was characterized in whole bodies of adult riceland prawns, Macrobrachium lanchesteri, based on substrate preference and inhibitor sensitivity. M. lanchesteri ChE activity was mainly attributable to acetylcholinesterase (AChE) since it preferentially hydrolyzed an AChE-specific substrate, acetylthiocholine iodide, over s-butyrylthiocholine iodide and was sensitive to 1,5-bis-(4-allyldimethyl-ammoniumphenyl)-pentan-3-one dibromide, a specific inhibitor for AChE. The effect of chlorpyrifos exposure on M. lanchesteri were also investigated. The 24, 48, 72 and 96 h LC50 values for chlorpyrifos were 3.37, 2.76, 2.58 and 2.53 μg L−1, respectively. Based on these values, adult M. lanchesteri were exposed to 0.5, 1.5, 2.5 and 3.5 μg chlorpyrifos L−1 for 24, 48, 72 and 96 h. Chlorpyrifos appeared to induce drastic enzymatic responses in M. lanchesteri. AChE activity was inhibited after 24 h of exposure in a dose- and time-dependent manner. The levels of lipid peroxidation increased significantly after 24 h of exposure. However, the activities of the antioxidant enzyme, catalase, and the detoxification enzyme, glutathione S-transferase, were reduced. In conclusion, M. lanchesteri is sensitive to chlorpyrifos and can serve as a bioindicator species for freshwater environmental assessment.

Isolation, characterization and virulence of the culturable bacteria from entire tissues of larval Ostrinia nubilalis (Hübner) (Lepidoptera: Pyralidae) were studied to obtain new microbes for biological control. A total of 16 bacteria were isolated from living and dead larvae collected from different maize fields in the Eastern Black Sea Region of Turkey. The bacterial microbiota of O. nubilalis were identified as Pseudomonas aeruginosa (On1), Brevundimonas aurantiaca (On2), Chryseobacterium formosense (On3), Acinetobacter sp. (On4), Microbacterium thalassium (On5), Bacillus megaterium (On6), Serratia sp. (On7), Ochrobactrum sp. (On8), Variovorax paradoxus (On9), Corynebacterium glutamicum (On10), Paenibacillus sp. (On11), Alcaligenes faecalis (On12), Microbacterium testaceum (On13), Leucobacter sp. (On14), Leucobacter sp. (On15) and Serratia marcescens (On16) based on their morphological and biochemical characteristics. A partial sequence of the 16S rRNA gene was also determined to confirm strain identification. The highest insecticidal activities were obtained from P. aeruginosa On1 (80%), Serratia sp. On7 (60%), V. paradoxus On9 (50%) and S. marcescens On16 (50%) against larvae 14 days after treatment (p < 0.05). Also, the highest activity from previously isolated Bacillus species was observed from Bacillus thuringiensis subsp. tenebrionis Xd3 with 80% mortality within the same period (p < 0.05). Our results indicate that P. aeruginosa On1, Serratia sp. On7, V. paradoxus On9, S. marcescens On16 and B. thuringiensis subsp. tenebrionis Xd3 show potential for biocontrol of O. nubilalis.

Simulium degrangei Dorier & Grenier, 1960 was recorded in southern and central Europe and in the Crimea and Caucasus. Its distribution pattern is scattered. The variability of the mtDNA gene encoding cytochrome oxidase I was studied in populations of S. degrangei from the Western Carpathians in Slovakia and in the northern Hellenides of Greece. In the analyzed samples, 21 haplotypes were recorded, of which twenty were private and occurred in only one mountain range, but one haplotype was found in all three studied populations from the Western Carpathians. Both haplotypes from the Hellenides were private, but these were not isolated and they fit into the haplotype network of the Western Carpathians. Statistics of genetic variability, different designs of analysis of molecular variation with non-significant differences and the pair-wise genetic distances support the hypothesis that there are no differences between the analyzed populations. Similarly, using nested clade phylogeographical analysis, no genetic-geographical structure was found.

Comparisons of morphology, nuclear gene sequencing and microsatellites were used to identify 19 hybrids between the related species Cottus poecilopus and Cottus gobio in three rivers of the Odra River drainage basin. All hybrids were the results of backcrossing and no F1 generation hybrid was found. The resulting progeny are fertile and continue in backcrossing with parental conspecifics. A high representation of backcrosses appears to indicate a lack of reproduction barriers within the hybridization process. It was impossible to differentiate individual back-cross categories on the sole basis of six loci. Mitochondrial haplotypes indicate that this is not a one-way process. In our study, hybridization was asymmetrical in favour of Cottus poecilopus.

To identify a simple indicator for detection of follicular vitellogenesis stage of cultured Caspian brown trout, the changes of plasma calcium and estradiol-17β along with ovarian histology were investigated over a reproductive cycle. For this purpose, the blood and gonad samples were taken monthly over a six months period. According to results, the concentrations of plasma calcium increased significantly during October and November but then declined steadily until the end of experiment. Estradiol-17β concentrations were low throughout the experiment, a clear peak was observed in October. Histological observations showed that all examined females were in vitellogenesis when the peak of plasma calcium and estradiol-17β were observed. Also, significant positive relationship was found between plasma calcium and estradiol. The good fitness of plasma calcium and estradiol-17β peak with histological observations suggests that plasma calcium could be a good and simple indicator for detection of follicular vitellogenesis stage in the cultured Caspian brown trout.

In the Indian subcontinent there is hardly any study that compares the bird community structure of urban/suburban areas with those of forest habitat. The present survey identified diverse assemblages of birds in the Pauri district at different elevations. A total of 125 bird species belonging to 40 families including two least count species (Lophura leucomelanos and Pucrasia marcolopha) were recorded during this survey in the forest and urbanized habitats of Pauri District (Garhwal Hiamalaya) of Uttarakhand state, India. The high elevation (Pauri 1600–2100 m a.s.l.), mid elevation (Srikot-Khanda 900–1300 m a.s.l.) and low elevation (Srinagar 500–900 m a.s.l.) contributed 88.8%, 63.2% and 58.4% of the total species respectively. Rarefaction analysis and Shannon diversity index showed that the high elevation forest habitat had highest bird species richness (BSR) and bird species diversity (BSD) followed by the mid and then the low elevation forests. BSR and BSD fluctuated across seasons at all elevations but not across habitat types. Present study provides a base line data about avian community composition in urbanized and natural habitats along altitudinal gradient in the study area. This information may be useful to the conservation biologists for the better management and conservation of the avifauna in the Western Himalaya, a part of one of the hot biodiversity spots of the world.

Somatic features (morphometric or meristic features, i.e., body weight, length of body, tail, hind foot and ear) and reproduction characteristics (number of embryos in uterus, reproduction potential, duration of reproduction season) are the basic attributes of the biology of a species. In the present paper, biometry of somatic characteristics and reproduction potential of two mountain vole species, the Tatra vole (Microtus tatricus) and the snow vole (Chionomys nivalis) which originate from various sites of Slovakia, were investigated. Among somatic features, the body length, tail length, hind foot length, length of ear and body weight were assessed. The highest variability was found in body length and the lowest one in the hind foot length. These characteristics were evaluated separately for sub-adult and adult individuals, and males and females. Values of somatic features and body weight of both species are larger in males than in females. Throughout the life of these rodents, the ratio between the length of tail and length of body does not vary considerably. The reproductive potential of the two species is, compared to other small mammals, very low and the length of reproductive season is shorter in the extreme conditions of the mountain environment. Thanks to a sufficient data set, it was possible to evaluate the influence of altitude on somatic features as well as on reproduction potential of M. tatricus and it was found that the average values of somatic features and body weight increased with increasing altitude.

Common features of embryonic genome activation in mammalian and non-mammalian embryos are the colocalization of pre-assembled complexes of maternally inherited nucleolar proteins, the so-called nucleolus precursor bodies and de novo synthesized transcripts with ribosomal DNA. The de novo transcription of messenger RNA and ribosomal RNA proteins is required for the development of functional nuclei during the major activation of the embryonic genome. The aim of our work was to investigate to what extent. Autoradiography and transmission electron microscopy has been applied in in vitro produced bovine embryos. The embryos were cultured to the late 8-cell stage with: α-amanitin; a specific inhibitor of RNA-polymerases II and III transcription; actinomycin D; a specific inhibitor of RNA polymerase I transcription; and without inhibitors (control group). Nucleoplasm and nucleolar structures displayed strong autoradiographic labeling and showed the initial development of fibrillo-granular nucleoli in the control group. In α-amanitin groups, however, in both inhibited groups of embryos, lack of autoradiographic labeling and disintegrated nucleolus precursor bodies stage were observed. Our study of α-amanitin as well as in actinomycin D groups proves inhibition of transformation nucleolus precursor bodies to active nucleoli. From our results follows, actinomycin D is able to penetrate through zona pellucida, what was shown for the first time.