Volume 70, Issue 2

A total of 152 halophilic bacteria were isolated from Yuncheng Salt Lake, China. Phylogenetic analysis based on 16S rRNA gene sequence comparisons revealed that thirty-four strains were related to the phylum Firmicutes and belonged to three families, Bacillaceae, Clostridiaceae and Staphylococcaceae. The other strains were identified as the members of Halomonadaceae and Idiomarinaceae, which belonged to the phylum γ-Proteobacteria. Nine strains showed <97% similarity of 16S rRNA gene sequence compared to other published species, which might represent novel species. The halophilic isolates exhibited various hydrolytic activities. A total of 74, 15, 70, 18, 23 and 3 strains were found to produce extracellular amylase, protease, lipase, cellulase, pectinase and DNAase, respectively. Most hydrolase-producers were members of the genus Halomonas. Combined hydrolytic activities were shown by some strains. Screening of antimicrobial activity indicated that 3, 6, 15, 12, 15 and 16 of halophilic isolates could inhibit Staphylococcus aureus, Escherichia coli, Candida albicans, Fusarium moniliforme, Fusarium semitectum and Fusarium oxysporum, respectively. Results from the present study indicated that halophilic bacteria may be developed as promising sources for novel biocatalysts or bioactive substances in the field of biotechnology.

In this study, a keratinolytic protease gene (named as kerD) from Aspergillus niger was cloned. The full-length coding sequence of kerD consists of 1,251 bp and encodes a protein with 416 amino acid residues with a molecular mass of 43,831 Da. A DNA fragment encoding mature kerD without its signal sequence was inserted into the expression vector pET30a(+) and successfully expressed in Escherichia coli. The recombinant protein was purified to approximately 100% purity using Ni-IDA affinity chromatography, and identified by Western blot. The recombinant enzyme had an optimal pH of 8.0 and was stable at pH 7.0-9.0. It exhibited an optimal temperature for activity of 70 °C and was stable at 30-50 °C. It was highly inhibited by 1,10-phenanthroline, ethylenediaminetetraacetic acid and sodium dodecyl sulphate, but activated by phenylmethanesulfonyl fluoride, Mg2+, Fe2+, Mn2+, Cu2+, Zn2+, Ca2+, dithiothreitol, Triton X-100, dimethyl sulfoxide and isopropyl alcohol. The recombinant enzyme could hydrolyse a broad range of protein substrates.

Effect of sucrose and its abscisic acid (ABA) dependence was studied for seed germination, growth and antioxidant related parameters in two wheat cultivars PBW343 (ABA-lesser sensitive) and C306 (ABA-higher sensitive) by using sucrose (Suc) with or without sodium tungstate (inhibitor for ABA biosynthesis) in culture medium. Exogenous Suc (at 1.5% and 3%) inhibited germination in both cultivars where ABA-dependency for such inhibitions was seen only under 3% sucrose in PBW343 and 1.5% sucrose in C306. Exogenous Suc at 1.5% stimulated growth, which was ABA-dependent in both cultivars. Exogenous Suc at 3% inhibited growth in both cultivars but such inhibitions were ABA-dependent only in PBW343. Suc supply (3%) decreased antioxidant enzymes and H 2 O 2 in PBW343 but increased these in C306. These changes were ABA-dependent. Suc supply (3%) increased reduced state of ascorbate in shoots and roots of PBW343 and in shoots of C306, and such increases were ABA-dependent. Suc-induced increases of proline were ABA-independent in both cultivars and were higher in C306.

The deleterious effects of soil salinity on biodiversity and agricultural productivity make it essential to devise strategies that would reduce its impact and expansion. For sustainable agriculture on dryland salinity affected areas, the use of native salt-tolerant plant species of economic value could provide a solution. In this work, the salinity tolerance potential of fifteen different Australian Acacia (wattle) species, pre-selected on the basis of phylogenetic relationships, were tested. The effects of salinity stress on morphological and physiological parameters, such as shoot and root lengths, shoot and root biomass, tissue water content, tolerance indices and accumulation of major cations (sodium, potassium, magnesium, calcium), all of which are indicators of salt stress response, were investigated. The species exhibited varied responses to the various parameters, some species consistently indicative of salt tolerance. Consolidating the tolerance indices based on morphological and physiological effects of salt stress, a method was developed to arbitrarily rank the species for their tolerance. Two highly tolerant, three tolerant and six moderately tolerant species could thus be identified. The work supports use of phylogenetics for initial screening for large genera, such as Acacia, and provides an experimental methodology for identification of candidate species for environmental stress tolerance and other applications.

To investigate the phylogenetic relationships among Elymus and related diploid genera, the genome donor of Elymus, and the evolutionary history of polyploid Elymus species, nuclear ribosomal internal transcribed spacer (ITS) sequences were analyzed for 10 Elymus species, together with 17 diploid taxa from 5 monogenomic genera. The phylogenetic analyses (Neighbor-Joining) supported two major clades (St and H). Sequence diversity and genealogical analysis suggested that (1) Elymus species were unambiguously closely related to Pseudoroegeria; (2) Pse. stipifolia might be serve as the St genome donor of polyploid Elymus species; (3) the Y genome might be originated from ancestral lineage of Pseudoroegneria (St); (4) the ITS sequences of Elymus were evolutionarily distinct and may clarify parental lineages and phylogenetic relationships in Elymus.

Lead (Pb), a widespread contaminant in terrestrial landscape, is highly detrimental to plant and animal life. Specifically, Pb-contaminated soils cause a sharp decrease in crop productivity, thereby posing a serious risk to agriculture. A study was planned to investigate the toxic effect of Pb 2+ (0, 16, 40 and 80 mg L −1 ) in the seedlings of maize (Zea mays), in terms of induced physiological and biochemical changes at initial hours of treatment (0-8 h). Increased accumulation of malondialdehyde (MDA) served as an indicator of cellular peroxidation. At 80 mg L −1 Pb 2+ , MDA content enhanced over the control by 175% after 2 h of exposure and increased further to 461% greater over the control after 8 h of exposure. Elevated superoxide ion (O −· 2 ) and H 2 O 2 contents suggested oxidative damage to the plants. The level of H 2 O 2 increased over control by 70%, 80%, 135% and 182% at 2, 4, 6, and 8 h after exposure to 16 mg L −1 Pb 2+ , respectively. In situ histochemical localization confirmed the level of lipid peroxides, increased accumulation of O −· 2 and loss of membrane integrity upon Pb 2+ treatment. Pb 2+ -induced oxidative stress triggered significant changes in the activities of antioxidant enzymes. A concentration-dependent increase was observed in the activities of the superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidase (GPX) and glutathione reductase (GR) in response to Pb 2+ , whereas catalases (CAT) was not able to provide protection against oxidative stress. These observations imply that Pb 2+ -induced oxidative stress during initial period (0-8 h) of exposure involved ROS accumulation and upregulation of scavenging enzymes except CAT as a defense against Pb 2+ -toxicity.

The interaction between zinc and cadmium was studied in tomato leaves (Solanum lycopersicum), in a hydroponic medium. Ten-day-old seedlings were treated with 25 μM CdCl 2 associated to different concentrations of ZnCl 2 (10, 50, 100, and 150 μM). We first observed that seven days after the metallic exposure Zn supply clearly reduced Cd concentration in leaves and simultaneously increased Zn uptake. Moreover, the addition of 10 μM ZnCl 2 relieved the leaves growth inhibition observed under Cd stress. At the same concentration (10 μM), Zn repaired the Cd damages on the membrane lipid content and fatty acids composition. In fact, Zn supplementation at low level (10 μM) restored and enhanced the quantities of the total lipid (TL), galactolipids (GL), phospholipids (PL) and neutral lipids (NL) that were reduced by 25 μM CdCl 2 . There was also a substantial amelioration in total lipid biosynthesis that was noticeably altered with Cd treatment. In 10 μM Zn-Cd treated leaves the [1-14C]acetate incorporation in total lipids as well as in all lipid classes was similar to that of control leaves. Likewise, levels of tri-unsaturated fatty acids that dropped in total lipids of Cd treated leaves showed an increase after supplementation of 10 μM Zn. Moreover, the beneficial effect of the same dose of Zn (10 μM) on Cd toxicity was confirmed by a significant decrease in the oxidative stress state which was marked by a reduction in thiobarbituric acid-reacting substances (TBARS) levels and a reduction in lipoxygenase (LOX, E.C.1.11.1.7) activity that was significantly enhanced by Cd treatment. These results suggest that 10 μM Zn supplementation proved to be inhibitory for Cd-induced oxidative stress and beneficial for the plant’s survival. However, higher Zn concentrations (50, 100 and 150 μM) and Cd are synergistic in their effect on plant growth parameters and lipid content.

Floodplain forests are sensitive to changes within the surrounding environment and contain the most highly invaded habitats. The overall aim of this study was to characterise floristic composition of floodplain forest along two different riparian corridors. The studied river ecosystems were influenced by human disturbances, but they have historically different hydrological management practices (e.g., damming and water regime management). We hypothesised that different hydrological management practices affect the composition and diversity of vegetation and influence multiple ecosystem functions and services in floodplain forests. A detailed study of the vascular plant species diversity of floodplain forests in the lower parts of two riparian corridors of the Thaya and Morava Rivers (South Moravia) was conducted. Altogether, 853 species of vascular plants were recorded, including 121 species of woody plants. We found 111 species that are protected by law or threatened according to the Czech Red List. We found 230 alien species, out of which 125 are archaeophytes and 105 are neophytes. Thirty-nine species are invasive; however, the most frequent group comprises naturalised archaeophytes (78). Differences in the effects of environmental factors on species richness and the proportions of alien and endangered species were tested using generalised linear models (GLMs). Differences in species composition in the two distinct riparian corridors were examined using non-metric multidimensional scaling (NMDS). Although only small differences were observed in the vegetation composition, we observed some differences in species richness and species composition between the riparian corridors. The most obvious difference was a higher proportion of alien species in the Morava River corridor than in the Thaya River corridor. In contrast, the proportion of endangered species richness was higher in the Thaya River corridor. We assume that the most probable explanation of the differences is the unique water management history for each river corridor.

The last version of Red List was published already more than 10 years ago but the knowledge of the flora of Slovakia has substantially improved due to large-scale mapping projects and taxonomical research. Therefore an updated fifth edition of the Red List is presented in this paper. For evaluation, we accepted IUCN categories (ER, CR, EN, VU, NT, LC, DD, NA, NE) and criteria (A-D), which were used first time in the history of Slovak Red Lists of ferns and flowering plants. In total 1 218 taxa are listed (the complete list of taxa is given in Electronic Appendix 1), which is nearly 34 % of the total number of 3 619 native taxa and archaeophytes of the flora of Slovakia. More than 80 taxa (7 %) were not confirmed in the flora (66 Regionally Extinct, 17 Probably Regionally Extinct), 155 taxa are evaluated as Critically Endangered (13 %), 171 Endangered (14 %) and 201 taxa are regarded Vulnerable (17 %). The largest group represent 347 taxa evaluated as Near Threatened (29 %), 162 taxa are assessed as Least Concern (13 %), 91 taxa are in category Data Deficient (7 %), and 8 taxa belong to categories of Not Evaluated and Not Applicable (1 %). In this Red List, 27 taxa are included as new for the flora of Slovakia and 14 taxa were rediscovered. Short history of Slovak Red lists of rare and threatened plants is also briefly mentioned.

Distribution pattern, prevalence and intensity of parasitaemia of heteroxenous apicomplexan blood parasite Hemolivia mauritanica and its vector tick Hyalomma aegyptium have been studied in 264 tortoises (212 Testudo graeca and 52 T. horsfieldii) throughout the Iranian territory. In T. graeca the highest prevalence and intensity of parasitaemia for H. mauritanica were recorded in the temperate mid and northern parts of the Iranian plateau, while the lowest values were found in the arid central part of Iran. No Hemolivia-positive samples were found in T. horsfieldii. Hyalomma aegyptium ticks were diagnosed in both tortoise species. The highest prevalence and intensity of infestation with H. aegyptium were recorded in western part of Iran, where climatic conditions are not so arid compared to central and eastern parts. No significant differences were found in intensity of parasitaemia of H. mauritanica between males and females. Similarly, no differences in intensity of parasitaemia were detected between different age groups (adults; more than 10 years, young; 5-10 years and juveniles; less that 5 years). Prevalence of H. mauritanica and H. aegyptium in adults was higher than in young and in juveniles. While intensity of infestation with H. aegyptium was significantly higher in adult tortoises compare to juveniles, no significant difference were found between intensity of H. aegyptium infestation in male and female tortoises.

The genus Ledermuelleriopsis Willmann, 1952 is recorded for the first time in South America. Ledermuelleriopsis verricula is redescribed based on material from Rio de Janeiro, Brazil.

The influence of agricultural management form on the structure and the species diversity of ground beetle (Carabidae) assemblages was studied. The research was carried out in the vicinity of Očová village (Central Slovakia) in 2005-2006. Carabid beetles were pitfall-trapped in four pairs of plots with the same crop, but conventionally and naturefriendly managed. In total, 54,376 individuals of 68 species of 30 genera were collected. The significantly positive effect of nature-friendly management on the number of carabid beetle individuals and species diversity was observed only in the plots with arable soil. Carabid beetles reached a higher species diversity in the plots with a higher pH and humus content. These parameters were typical of nature-friendly managed fields, and of the conventionally managed permanent grass stands. In general, number of carabid beetle individuals and species was higher in arable soils than in permanent grass stands.

The plant Jurinea cyanoides (L.) Reichenbach (Asteraceae), protected under the European Commission Directive on the conservation of natural habitats and of wild fauna and flora (92/43/EEC), is critically endangered in Central Europe. The centre of its continuous range of distribution is in Ukraine and in a part of European Russia. Natural isolated populations have refugial distribution in the Czech Republic and Germany. Interactions of insects from different feeding guilds (i.e., phytophages, pollinators, parasitoids, predators) with J. cyanoides were studied in central Ukraine, Czech Republic and Germany. The insect community identified on J. cyanoides and differences in its composition on robust populations of plant in contrast to sparsely populated periphery of its range are discussed. Altogether 78 species belonging to six orders (Coleoptera, Diptera, Hemiptera, Hymenoptera, Lepidoptera and Neuroptera) were recorded. Larvae of the pyralid moth (genus Dioryctria), tephritid fruit flies (Acanthiophilus helianthi), and several polyphagous Heteroptera species are identified as main taxa feeding on inflorescences and seeds of J. cyanoides, with a potential to reduce the plant’s sexual reproduction. The impact of phytophagous insects is, however, considered only a secondary reason for the decline of the Czech populations of J. cyanoides on which a fairly low number of insect species were recorded in comparison with Ukraine and Germany; habitat loss and inbreeding effect are probably major negative factors. Several new host plant-insect and host-parasitoid associations are reported for insects on J. cyanoides.

On the basis of several preclinical and clinical studies, we have assumed that phytochemicals may play an important role in plasma lipid metabolism. The aim of this study was to evaluate the effects of dietary administered Chlorella pyrenoidosa, young barley and fruit peel polyphenols from Flavin7 on plasma lipid metabolism in breast cancer model in female rats. The phytopharmaceuticals were dietary administered at two different concentrations (0.3% and 3%). The administration of drugs lasted for the whole duration of the experiment (14-15 weeks) until autopsy. At the end of the experiments blood was collected from animals and serum lipid parameters were evaluated. Flavin7 in lower dose significantly decreased LDL-cholesterol and in higher dose significantly decreased triacylglycerol and VLDL-cholesterol levels. Chlorella at the higher dose significantly decreased LDL- and VLDL-cholesterol and triacylglycerol levels. Young barley significantly decreased LDL cholesterol (at the higher dose); on the other hand, a tendency of increased serum triacylglycerol and VLDLcholesterol levels was found (in both doses). Our results pointed to significant beneficial effects of fruit peel polyphenols from Flavin7 and C. pyrenoidosa on plasma lipid metabolism in female rats. Further research is needed to elucidate the health benefits of phytochemicals in whole foods.

Diabetes mellitus (DM) is considered as a risk of cardiovascular diseases due to the expression of certain pathological factors, including hormones, such as serotonin. The aim of the current study was to evaluate the bridging role of serotonin between DM and cardiac valvulopathy via reactive oxygen species (ROS) generation. Eighty diabetic patients and 50 healthy individuals along with experimental models were studied. DM was induced in the rat model via intraperitoneal alloxan injection and subsequently treated with serotonin. ROS, superoxide dismutase and brain natriuretic peptide (BNP) expressions analyses as well as histopathologies were also performed. An increase in ROS concentration and down-regulation of antioxidant enzyme activities in diabetic patients as well as serotonin-treated diabetic experimental models were observed. The heart valves of the serotonin-treated diabetic rats were impaired along with BNP expression when compared to the normal subjects. The current study suggested that endogenous enhanced serotonin level may contribute as a bridge between diabetes and cardiovascular diseases via acceleration of free radical generation in diabetic conditions.

Parasitological studies were carried out on pulli (n = 20) collected on a breeding colony near the Włocławski Reservoir on the Vistula River (Central Poland). Larvae of the nematode Contracaecum rudolphii (mean intensity of infection 9.3) were detected in the stomachs of all chicks, with adult worms present in two chicks. Larvae of the nematode Porrocaecum ardeae (mean intensity 3.1), the tapeworm Paradilepis scolecina (58.5) and flukes of five genera: Tylodelphys, Posthodiplostomum, Apharyngostrigea, Paryphostomum and Echinochasmus (mean intensity in total 28.1) were found in the intestines of all chicks. The studies demonstrated that grey heron chicks are infected with parasites very early in life, which may adversely impact their health. The presence of adult C. rudolphii nematodes indicates a transfer of parasites from adult birds to chicks during feeding.