Volume 68, Issue 1

In the present study the biofilm-forming characteristics of 99 serotyped (DMC strains) and 41 genus level-identified (IS strains) Salmonella strains originating from Turkey were investigated. The strains were selected based on their ability to show the biofilm morphotype on Congo red agar plates. In addition, all strains were evaluated with regard to properties related to forming pellicle structures, physical differences of pellicles, any changes in the media associated with the formation of pellicles, and the presence of cellulose within the formed biofilm matrix as determined using 366 nm UV light. The Salmonella Typhimurium DMC4 strain was the best producer of biofilm grown on polystyrene microtiter plates (optical density at 595 nm: 3.418). In subsequent experiments industrial process conditions were used to investigate different morphotyped Salmonella strains’ biofilm-forming capability on stainless steel, a commonly preferred surface for the food industries, and on polystyrene surfaces. The effect of other important industrial conditions, such as temperature (5, 20, 37°C), pH (4.5, 5.5, 6.5, 7.4) and NaCl concentration (0.5, 1.5, 5.5, 10.5%) on the production of biofilm of the different morphotyped Salmonella strains (DMC4; red, dry and rough morphotyped S. Typhimurium, DMC12; brown, dry and rough morphotyped S. Infantis, DMC13; pink, dry and rough morphotyped S. subsp. Roughform) were also assessed. On the other hand, pH values exhibited variable effects on biofilm-forming features for different Salmonella strains on both polystyrene and stainless steel surfaces.

The incidence of infections due to organisms resistant to β-lactam antibiotics has increased sharply in recent years. The goal of this study was to investigate the β-lactam resistance in 151 Escherichia coli strains isolated from chickens over a two-year period. Extended spectrum β-lactamases (ESBLs) were present in 24 strains (16%), ESBL phenotype was identified by interpretative reading of minimal inhibitory concentration values of ceftriaxon (CRT ≥ 7.1 mg/L), ceftazidime (CAZ ≥ 3.4 mg/L) and ceftiofur (CFF ≥ 8.7 mg/L). PCR detection revealed the presence of the bla CMY-2 gene and CTX-M-1 group. We detected high resistance to ampicillin (92%), streptomycin (63%), tetracyclin (70%), ceftiofur (40%), floroquinolones (enrofloxacin 68%, ciprofloxacin 62%), florfenicol (18%), chloramphenicol (21%) and cotrimoxazol (43%). We also investigated the presence of virulence factors and mobile genetic elements, and performed plasmid replicon typing in 24 selected strains. The most prevalent integrase among the isolates was the integrase 1 with gene cassettes dfrA, aadA and genes sul1 and sul2. Plasmid mediated quinolone resistances (qnrS) were also detected in two strains. Plasmid typing showed that the Y and IncI1 were dominant plasmid replicons. The genes iss, kpsII, tsh, iutA were the most frequently detected virulence genes in ESBL-positive strains. These results demonstrate that broilers in Slovakian food markets and farms could be the source of ESBL-producing E. coli, as well as virulent and resistant strains representing a potential risk for the human population.

In the present study we focused on the impact of macrophyte cover (composed mainly of the Lemna genus) on phytoplankton taxonomic and functional diversity. Some important environmental parameters, mainly light (KdPAR), and the chemical conditions (pH, dissolved oxygen, ammonium, soluble and total forms of phosphorus) were closely related to the pleustophyte cover. Among them, the key factor in the phytoplankton ecology of the studied oxbow lakes was the dense macrophyte cover which strongly reduced the illumination of water. Neither differences in the mean nutrient concentrations between the lakes with FFP (Free Floating Plants) absence and those with FFP dominance nor significant relationships between nutrients and the phytoplankton structure were observed. The species composition of phytoplankton and the functional (FG) and morpho-functional (MFG) groups reflected the differences between the habitats connected with hydromacrophytes. The free-floating macrophyte cover favours mixotrophic and heterotrophic species, mainly Euglenophyta (coda W1 and W2) and chrysophytes (codon Ws) as well as shade-adapted cyanobacteria with the high tolerance of the low oxygen content (codon K). In lakes with FFP absence — taxa from Chlorophyta and Bacillariophyceae (associations X1, J, and D), or filamentous cyanobacteria (codon S1) dominated the phytoplankton. MFG were less related to the oxbow type and exhibited greater similarity between lakes independently of the presence or absence of FFP. Only unicellular Cyanoprokaryota which created MFG 4 and colonial chroococcales (MFG 5b and 5c) reached a greater percentage share, especially in oxbow lakes with FFP dominance.

Ranunculus cabrerensis is an endemic and endangered species of the Northwestern Iberian Peninsula. The molecular markers AFLP and ISSR were used to investigate the genetic diversity and population structure of four populations across its known distribution. Fifteen selective primer combinations of AFLP and seventeen ISSR primer combinations produced a total of 2830 and 103 unambiguously repeatable fragments respectively, of which 97.57 and 81.38% were polymorphic for both markers. The genetic diversity of R. cabrerensis at species level was high (H E = 0.294 by ISSR and H E = 0.191 by AFLP) and differentiation between sampled locations was also relatively high (G ST = 0.316 and 0.158 by ISSR and AFLP analysis respectively) compared to other studies of endangered and rare species using the same techniques. The analysis of molecular variance (AMOVA) indicated that the main genetic variation was within sampled locations (73% by AFLP; 52% by ISSR), even though the variation among locations was also significant. Principal Coordinates, NeighborNet and Bayesian analyses revealed a weak but significant relationship between the genetic structures of different populations in R. cabrerensis, with gene flow acting as a homogenizing force that prevents stronger differentiation of populations. Finally, suggestions for conservation strategies to preserve the genetic resources of this species are outlined.

Jatropha curcas (jatropha) is a multipurpose plant with potential as a raw material for biofuel. In the present study, a total of 43,349 expressed sequence tags (ESTs) from J. curcas were searched for type and frequency of simple sequence repeat (SSR) markers. Five thousand one hundred and seventy-five sequences were indentified to contain 6,108 SSRs with 90.8% simple and 9.2% compound repeat motifs. One hundred and sixty-three EST-SSRs were developed and used to evaluate the transferability and genetic relatedness among 4 accessions of J. curcas from China, Mexico, Thailand and Vietnam; 5 accessions of congeneric species, viz. J. gossypiifolia, dwarf J. integerrima, normal J. integerrima, J. multifida, J. podagrica; and Ricinus communis. The polymorphic markers showed 75.56–85.19% transferability among four species of Jatropha and 26.67% transferability across genera in Ricinus communis. Investigation of genetic relatedness showed that J. curcas and J. integerrima are closely related. EST-SSRs used in this study demonstrate a high efficiency of cross species/genera amplification and are useful for identifying genetic diversity of jatropha and its close taxa and to choose the desired related species for wide crossing to improve new varieties of jatropha. The markers can also be further exploited for genetic resource management and genetic improvement of related species/genera through marker-assisted breeding programs.

A cytotaxonomical description of Cyclopogon (Spiranthinae, Orchidaceae) is carried out through a deep karyotype analysis of four species from NE Argentina. Distinctive karyotype parameters concerning the chromosomes number, morphology, size and symmetry and the genome size associate to each taxon. Cyclopogon calophyllus (2n = 2x =28; 18m + 10sm), C. congestus (2n = 2x = 32; 26m + 6sm), C. elatus (2n = 2x = 28; 18m + 10sm) and C. oliganthus (2n = 4x = 64; 40m + 24sm) possess symmetrical karyotypes (i-mean = 40.01–42.84; A 1 = 0.24–032; r>2 = 0.06–0.29) and excluding C. congestus (A 2 = 0.26; R = 2.62) unimodality is the rule (A 2 = 0.12–0.20; R = 1.73–1.92). Diploid taxa show a terminal macrosatellite in the m pair no. 2 (large arm) and share a comparable mean chromosome length (ca. 2.75 μm) and genome size (ca. 40 μm), superior to the tetraploid C. oliganthus (ca. 2 and 32 μm, respectively). The novel data added to preceding cytological, morphological and molecular approaches involving Cyclopogon and those related taxa of Spiranthinae largely based on x = 23 support the hypothesis that the unusual 2n and the karyotype morphology of Cyclopogon is an evolutionary advance within Spiranthinae with a basic reduction to x = 14 or 16 by chromosome fusions. A polyploid-dysploid series added to dibasic hybridization explain the extant 2n diversity though a paleopolyploid series on x = 7–8 is also possible.

The frequency of parasitism of the European mistletoe, Viscum album L. subsp. album, in the city of Lodz, a typical major city in Poland, was investigated. The infection prevalence and intensity of the mistletoe Viscum album subsp. album on its main host, Acer saccharinum as a function of host size was also investigated. The parasite showed a strong preference for alien, planted tree species (i.e. A. saccharinum, Populus×canadensis and Robinia pseudoacacia). In 2009–2011, V. album subsp. album was observed on 28 host taxa of trees and shrubs, which represents the highest diversity of host trees in a single locality in the Poland. Within the studied area 2147 trees were infested by mistletoe. The distribution of mistletoes (V. album subsp. album) among A. saccharinum hosts is significantly aggregated. The intensity of mistletoe infection in the silver maple trees was affected by the individual tree characteristics, such as the height of the tree. The overall level of aggregation as indicated by the variance to mean ratio of mistletoe numbers per host fell within the midrange of values found in other published studies of host-mistletoe interactions. The higher mistletoe infection prevalence in taller trees results from differential dispersal of mistletoe seeds to tall trees as well as differential survival of established mistletoes on tall trees. The incidence of mistletoe was higher in city centre (zone of high density development) than it was on the outskirts of a city (outer marginal zone). It was found that the abundant occurrence of mistletoe was recorded in the stands of increased nitrogen input, while other stands have little or no mistletoe infection present. Thus, this mistletoe species uses both passive and active uptake, which may be a selective advantage in a nutrient-poor environment or on a nutrient-deficient host species.

Melampyrum pratense is one of the most successful root-hemiparasitic species of temperate Europe occurring in various habitats including both forest and open communities. The species shares its life history traits (large seeds, annuality, lack of clonality) with most of other hemiparasitic Orobanchaceae. Due to lack of vegetative propagation, the reproductive success of a M. pratense individual largely depends on the seed production. We used an individual-based observation of ontogenetic development of plants and analysed the development of spatial distribution of individuals to reveal links between fecundity of individuals and their vegetative growth in the context of population dynamics within one growing season. We demonstrated a tight dependency of seed production on vegetative growth and survival till the end of the growing season. Plants that were still alive and big (due to a high number of lateral branches) in the end of August produced the highest numbers of seeds. These were several times higher than the population average due to positively skewed distribution of seed production across individuals. The rate of premature mortality was rather low (below 50%) once seedlings successfully emerged and was most likely caused by intraspecific competition. By contrast, moderate level of herbivory (stem clipping by grazers) had a limited effect on the survival and the inflicted biomass losses seemed compensated. Therefore, despite being an annual, M. pratense appears well-adapted to its life in perennial-dominated forest communities, which is also underpinned by its hemiparasitic strategy providing essential resources and allowing to avoid below-ground competition.

Cadmium (Cd) has been identified as a significant pollutant due to its high solubility in water and soil and high toxicity to plants and animals. Rice, as one of the most important food crops, is grown in soils with variable levels of Cd and therefore, is important to discriminate the Cd tolerance of different rice cultivars to determine their suitability for cultivation in Cd-contaminated soils. This study investigates the primary mechanisms employed by four rice cultivars in attaining Cd tolerance. HA63 cultivar reduces Cd uptake by increasing Fe absorption through activation of phytosiderophores. T3028 cultivar accumulates the highest level of Cd in leaves while also activating its reactive oxygen species (ROS) scavenging system, including antioxidant enzymes and phytochelatins. In some rice cultivars (such as HA63), a cyanide-resistant respiration mechanism, important in Cd detoxification, was also promoted under the Cd stress. In conclusion, different rice cultivars may adopt different biochemical strategies and respond with different efficiency to Cd stress.

A rice lesion mimic mutant, lm3, was obtained by the mutagenesis of an indica cultivar, 93-11, using γ-ray radiation. Brownish lesions appeared on the leaves of lm3 at the young seedling stage and persisted until the ripening stage. The lm3 mutant was characterised by a shorter plant height and delayed heading compared with the wild-type 93-11. A genetic analysis indicated that the lesion mimic phenotype was controlled by a single recessive gene. Using simple sequence repeat (SSR) markers, the target gene LM3 was first located between marker RM5748 and RM14906 on chromosome 3. We then developed Insertion-Deletion (InDel) markers to fine-map LM3, and the locus was localised to a 29 kb region defined by two InDel markers, In12571 and In12600. Five ORFs were predicted in the candidate region, and DNA sequencing detected a single-nucleotide polymorphism (SNP) in the coding region of LOC Os03g21900. The SNP in the fourth exon (C in 93-11; T in lm3) of LOC_Os03g21900 results in the substitution of a proline (P) with a serine (S) at the 140th amino acid of the deduced uroporphyrinogen decarboxylase protein. We did not detect polymorphisms in the other predicted ORF regions between lm3 and 93-11. These results suggest that LOC_Os03g21900 is the most likely candidate gene for LM3.

2-C-methyl-D-erythritol 4-phosphate cytidyltransferase (MCT) catalyzes the third reaction in the plastidial non-mevalonate pathway, which provides the precursors for ajmalicine. A full-length cDNA encoding MCT (RvMCT) was identified from hairy roots of Rauvolfia verticillata. The full-length 1,499-bp cDNA of RvMCT had a 945-bp coding sequence that encoded a 314-amino-acid protein with an N-terminal chloroplast transit peptide of 67 amino acid residues. RvMCT exhibited homology with other plant MCTs at the levels of sequence and structure. The phylogenetic analysis revealed the plant MCTs could be divided into three separated clusters including gymnosperms, monocotyledons and dicotyledons. Gene expression of ajmalicine metabolism (DXR, MCT, MECS, HDS, HDR, STR and SGD) in hairy roots, roots, stems, old leaves, young leaves and barks was analyzed by quantitative PCR. All the seven genes had higher expression levels in hairy roots than in other plant organs. This suggested hairy roots of R. verticillata possessed more active alkaloid metabolism than other organs and it was the reason that hairy roots produced higher levels of ajmalicine. Furthermore, the expression of DXR, MECS, HDS, HDR, STR and SGD genes was not detected in stems (only MCT detected in stems), so it could be presumed that stem acted as a transporter tissue of ajmalicine. Finally, the colour complementation assay indicated that the function of RvMCT was the same as Arabidopsis MCT. Molecular cloning, characterization and functional identification of RvMCT will be helpful to understand more about the role of MCT involved in ajmalicine biosynthesis at the molecular level.

Beneficial effects of whole grains of cereals and pseudocereals and their fractions to human physiology are well known and broadly published. Especially secondary metabolites, dominantly from the category of phenolics (or polyphenols), beneficially influence the health physiology and/or prevent disease progress. Within the frame of this study, ten genotypes of four cereals or pseudocereals, respectively, were chosen for their antioxidant activity, determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power (FRAP) and β-carotene-linoleic acid bleaching model (BCLM) mechanisms. Tested genotypes were selected from primary collection based on their antioxidant activity values, as well as higher level of flavonoids or phenolic acids. The stability of antioxidant properties after thermic, acidic, and basic treatments was evaluated. The oat cultivar Sirene and buckwheat cultivar Bogatyr expressed high level of the antioxidant activity, but they lost it due to all types of treatment. Oppositely, treatments increased antioxidant activities in some samples, especially in oat cultivar Maris Oberon, wheat cultivar Ines and Karolinum, or partially in barley cultivars Kompakt (after basic treatment) and Jubilant (acidic and basic treatments). The lack of the antioxidant activity could be observed due to destruction of the key compounds responsible for the antioxidant effect, whereas the increasing activity could be seen due to release of the aglycons from glycosidic forms after treatment. The stability of antioxidant properties could be a valuable parameter of the raw material for manufacturing special foods with functional properties.

In this study pro/anti-oxidant activities of Thymus revolutus Célak (T. revolutus C.) essential oil and its two main components on Hep G2 cells were determined. Twelve components were identified in T. revolutus C. essential oil by GC and GC-MS and the main components of the oil were cymene (32.57%) and γ-terpinene (17.18%). The essential oil showed more cytotoxicity than its two main components on Hep G2 after 24, 48 and 72 hours incubations. Membrane damage effects of the essential oil and its two main components on Hep G2 cells were increased by incubation with IC10, IC50 and IC70 concentrations. Lower essential oil (<IC50) concentrations had cytoprotective and membrane protective effects against H2O2. The free radical scavenging activity of the essential oil of T. revolutus C. (EC50 = 250±0.2 μg/mL) was superior to its two main components (neither not active). Oxidation of linoleic acid was effectively inhibited by T. revolutus C. (72.8%), while inhibition values of linoleic acid oxidation were calculated as 23.4.% and 26.7% for cymene and Γ-terpinene, respectively. These findings suggest that essential oils and its two main components had both pro-oxidant and protective (antioxidant) effects and they exhibited those effects depending on concentration.

First detailed limnological study was performed from April 2003 to June 2008 in a 396.6 km long section of the Serbian Danube (divided in three parts; from 1260 r-km to 863.4 r-km) to examine community composition and spatial distribution of Mollusca with special attention to the expansion of Ponto-Caspian species, propagation of invasive and introduced species and occurrence of endemic species. Mollusca samples were collected at 15 sites in April, June, September and November. We investigated the spatial distributions and calculated the faunistic similarity of localities with respect to the community composition using the Sørensen Quotient of Similarity. Site variation in analyzed qualitative traits was examined using correspondence analysis. Additionally, the obtained Complete Linkage City-block (Manhattan) distances among sites/parts were subjected to UPGMA (unweighted pair-group method using arithmetic averages) cluster analysis. The class Gastropoda was represented by 18 species from two subclasses (Prosobranchia — six families and Pulmonata — three families).The class Bivalvia was represented by 15 species from four families of order Eulamellibranchiata. Mollusca were represented by 33 species belonging to 17 genera and 13 families. Out of five recorded Ponto-Caspian species in the studied Danube stretch, Lithoglyphus naticoides (Pfeiffer, 1828) and Dreissena polymorpha (Pallas, 1771) are assumed to be invasive species with the highest occurrence frequency (F = 1, each). Four new species in invertebrate fauna for the Danube, denoted as the introduced species — Neozoa, were identified: Theodoxus fluviatilis (L., 1758), Corbicula fluminalis (Müller, 1774), C. fluminea (Müller, 1774), and Sinanodonta woodiana (Lea, 1834). The only endemic species of Gastropoda found in the Danube was Viviparus acerosus Bourguignat, 1862.

For the first time the life cycle of the common land snail Trochulus hispidus was completely described in Central Europe (Poland). This is a semelparous species predominantly with an annual life cycle and the reproductive period lasting from April till October. The first young snails hatch in spring, grow rapidly in summer and reach ca. 4 whorls until winter. In spring of the next year they mature and reproduce. After that they die. There is hardly any growth from late autumn till early spring. The average proportional growth rate is ca. 0.3 whorl/month in the wild. The fastest growth is present in the youngest snails and then gradually decreases over the course of their age. Laboratory and field observations allowed for establishing the following life cycle parameters: eggs calcified, almost spherical, ca. 1.5 mm, laid in spring and summer in batches of between 1 and 47. Time to hatching is 6–24 days, hatching is asynchronous; newly-hatched snails have approximately 1.5 whorls. Analysis of food preferences revealed, that T. hispidus tends to restrict its diet during the life. Generally the youngest snails equally consumed leaves of all four tree species offered (Fraxinus excelsior, Acer pseudoplatanus, Tilia cordata and A. platanoides) whereas adults preferred F. excelsior over A. pseudoplatanus and A. platanoides.

Spatial patterns in macroinvertebrate communities and some abiotic factors were examined in three rheocrene springs and their springbrooks (Kraków-Częstochowa Upland, southern Poland). The mean discharge of particular springs ranged from 5 to 11 L s−1, and its annual fluctuations were small. Water temperature was very stable at all sampling sites. In the eucrenon the number of benthic taxa was the smallest (9–14 determined to the family level), but the densities were the highest (approx. 14000 ind. m−2). The biggest changes in macroinvertebrate composition were observed in the modified hypocrenon, which is an artificial pond. The lowest number of taxa were found in a natural, short springbrook with a nondiversified bottom substrate. The density of crenophilic taxa (Drusus trifidus, Dugesia gonocephala, Elmidae) diminished along the springbrooks, while the opposite trend was observed for ubiquitous taxa (some Oligochaeta, Asellus aquaticus and Chironomidae). Even in a very short natural springbrook (30 m), Drusus trifidus, the only species of Trichoptera found in the springs discussed here, goes through the entire development cycle. The strongest influence of a big river was observed at the outflow of one of the natural springbrooks, where the highest number of riverine oligochaete species were found. The benthic fauna of the springs studied here differed from that found in other springs in this area — the absence of the typical crenophilic species Bithynella austriaca (Gastropoda) and the presence of Gianus aquedulcis (Oligochaeta) may indicate the autonomy of the spring fauna in the Mstów area, possibly resulting from the postglacial geomorphological formation of this region or differences in habitat conditions.

A new Uropodina genus (Sinharaja gen. n.) is described on the basis of two new species (S. ceylonensis sp. n. and S. sinhala sp. n.) from Sri Lanka. The new genus differs by the strongly sclerotised dorsal lines, pentagonal pygidial shield, the shape and length of dorsal setae and the shape of first and fourth hypstomal setae, internal malae and epistome from the other known Uropodina genera.

In 1993–1995 and 2009–2011, during investigation of reproduction biology of hoopoe, 63 nests of this species were collected after fledging of the chicks in Slovakia and Austria and their arthropod fauna was analyzed. Altogether 5,481 individuals and 34 species of mesostigmatic mites were found in 82.5% of the nests examined. The nidicolous mite Androlaelaps casalis was most abundant and frequent, representing 91.7% of all individuals. The richest in species were saprophilous mites (64.7% of all recorded species), while ectoparasites of the genera Dermanyssus and Ornithonyssus represented only 0.4% of all mites. Only 8 beetle species represented by 65 individuals were found in 18 nests. The dominant trophical group were carnivores (mainly nidicolous Gnathoncus buyssoni) with almost identical representation in (86.2%), followed by a similar representation of necrophags (10.8%). Unlike nests of other birds, the typical nidicolous species Haploglossa puncticollis and the fungivors were absent due to the dry character of studied countryside and placement of the nests in boxes situated in vineyard cottages. Only one species of fleas, Ceratophyllus gallinae — parasitizing first of all in the passeriform birds and being particularly abundant in the cavity nesting birds was recorded in hoopoe nests.

This study deals with the species distribution and structure of mite communities living in the fur of small mammals. Mesostigmatic mites (Acarina: Mesostigmata) of several small mammal species were studied at several sites of Slovakia situated in the Podunajská nížna lowland. The aims deal with ecological evaluation of mesostigmatic mites and their small mammal hosts. The research was done from 2005 to 2007. Several small mammal species were caught at 16 sites in the research area. These represent 13 types of biotopes. On the captured small mammals we recorded 3,141 individuals of mesostigmatic mites (7 families and 22 species). The dominance and the diversity of the recorded mites were analysed. Mites being interpreted as the variable one were evaluated by cluster analysis and multifactor analysis (detrended correspondence analysis, principal components analysis, redundancy analysis). Small terrestrial mammals as hosts of mites represent “holders” of environmental factors, in the analysis they are being evaluated as explanatory variable mainly. They transform direct impact of the environment and for that reason the ecological evaluation of mites is specific comparing to other taxonomical and ecological groups of animals.

We investigated a community of syntopically occurring horseshoe bats (Rhinolophus hipposideros, R. euryale, R. ferrumequinum) in southern Slovakia. The faecal pellets of these bat species were collected in the field and later analysed under a dissecting microscope. The three species studied are known to be very similar as far as their ecology, echolocation and preferred habitats are concerned, but they diverge significantly in their body sizes. In this study, all three species fed predominantly on moths [59–80 percentage frequency (%f); 87–95 percentage volume (%vol)], but their diet compositions differed in the size of individuals consumed. The smallest bat species (R. hipposideros) fed only on the smallest moths (%f = 59; %vol = 87), the medium-sized species (R. euryale) mainly on medium-sized moths (%f = 60; %vol = 74) and the largest one (R. ferrumequinum) especially on the largest moths (%f = 54; %vol = 89). Despite similar preferred habitat and the main prey category, the rates of trophic niche overlap were surprisingly low. The trophic niche percentage overlap was 7–31% (computed from %f data) and 1–20% (computed from %vol data), respectively and suggests an extraordinary importance of mere divergences of bats in their body sizes for trophic niche partitioning and stable species coexistence.