Volume 71, Issue 2

This paper delivers molecular biological study of antimicrobial resistance in clinical isolates of Campylobacter spp. in Slovakia. During the year 2015 a total of 828 Campylobacter strains collected from five regions in Slovakia were examined for resistance to ciprofloxacin (CIP), tetracycline (TET), erythromycin (ERY) and ampicillin with sulbactam (SAM). The highest incidence of Campylobacter resistance was observed for CIP (64.8%), followed by TET (30.5%) and SAM (26.3%). All strains were susceptible to ERY. Twenty selected antibiotic-resistant strains were examined for the presence Campylobacter β -lactamase OXA-61, multidrug efflux pump cmeB and TET tet (O) genes by PCR. From twenty CIP-resistant strains, the cmeB gene was present in 12, while the chromosomal OXA-61 β -lactamase occurred in 11 strains from 18 SAM-resistant Campylobacters, and tet (O) was present in 14 strains from 18 TET-resistant ones investigated. All three genes together occurred within a single strain in five Campylobacter jejuni and one Campylobacter coli strains isolated from eastern Slovakia. Clonal relatedness of strains was detected by a MALDI biotyper, which revealed two clusters of C. jejuni and one cluster of C. coli . The results showed that the epidemiologically important strains with high fluoroquinolone resistance occurred among the Campylobacter strains. An important finding is represented also by the evidence of the OXA-61 β -lactamase in more than one half of investigated strains, because SAM is frequently used for therapy in children and pregnant women.

Natural dyes and pigments are emerged as an important alternative to potentially harmful synthetic dyes. There is a growing demand for eco-friendly/non-toxic colourants specifically for textile dyeing because of its sustainability, green chemistry and improved eco-balances. The present study aimed to evaluate the dyeing potential of violet pigment from Chromobacterium violaceum UTM5. Dye-ability of violet pigment on different fabrics, i.e. pure cotton, pure silk, pure rayon, jacquard rayon, silk satin, cotton and polyester were tested using mordants and colour fastness tests were performed. Results from this study showed that violet pigment is capable of dyeing both natural and synthetic fibres. The use of slake lime mordant resulted in darker colour for cotton and silk satin. The colour fastness test showed fair to excellent (3/4) rating for fabrics dyed with violet pigment. This study demonstrates that violet pigment may have commercial demand in the textile industry.

The essential oil composition and in vitro antioxidant activity of eight important betel vine landraces, such as Balia, Chandrakala, Dandabalunga, Desibangla, Godibangla, Karpada local, Maghai and Nahua were evaluated in this research. GC and GC-MS analysis of the essential oil resulted in identification of 50 compounds representing 95.97% of the oil. Major compounds were eugenol, terpinolene, terpinene, cadinene, caryophyllene, 3-carene, etc. The plant essential oil was also subjected to screening for the evaluation of their antioxidant activities using 2,2-diphenyl-1-picrylhydrazyl assay. The highest antioxidant activity was shown by Balia landraces, whereas the lowest activity by Godibangla landraces. Essential oil showed a great variety of compounds in their chemical composition and antioxidant activity. This might be due to the affect of different environmental factors and soil nutrients of betel vine cultivating zones. The GC-MS chromatogram with antioxidant activity of the above betel vine landraces would not only help to select the elite chemotypes, but also to enhance their future industrial applications.

A specific and sensitive conventional and real-time PCR assays were developed for the detection and quantification of Rhizoctonia solani populations infecting pulses. Five different sets of R. solani specific molecular primers ARSF1&R1, ARSF2&R2 and ARSF3&R3, ARSF4&R4 and ARSF5&R5 were designed from the internal transcribed spacer sequences of R. solani isolates associated with pulse crops representing 7 anastomosis groups (AG1, AG2-2, AG2-2LP, AG2-3, AG3, AG4 and AG5). The designed primers yielded variable amplicons ranging from 174 to 563 bp specific to R. solani populations. The markers proved to be highly sensitive to R. solani and were able to detect the pathogen from both mycelium and infected mung bean plants at various concentrations of genomic DNA ranging from 0.025 to 1.0 ng using conventional PCR. The markers were also found suitable for real-time PCR assay with enhanced sensitivity up to 1.24 pg genomic DNA. The efficacy of markers for early detection of R. solani infection in mung bean plants was also tested and the marker ARSF4&R4 was the most sensitive in detecting the pathogen at 24 h after artificial inoculation under field and at 6 h after artificial inoculation under in vitro conditions. The radioactive and fluorescein amidite labelled probes for markers ARSF4&R4 and ARSF5&R5 were designed and tested for cross-reactivity and sensitivity by Southern blotting and real-time PCR analyses, respectively. Both probes proved to be highly sensitive and specific only to R. solani . Both quantitative PCR and probe-based diagnostics have been developed for R. solani detection in pulses.

Study describes protocols for in vitro propagation of Gentiana utriculosa L. via axillary shoot multiplication and indirect somatic embryogenesis. Shoot cultures were established from seedling epicotyl explants cultured on MS medium supplemented with 0.25 mg L −1 BA and 0.1 mg L −1 IAA. Medium containing 2% sucrose and 0.2 mg L −1 BA improved multiple shoot production, providing 2.3 shoots per explant. The highest rooting (29.6%) was obtained on medium with 1/2 MS mineral salts and 0.5 mg L −1 NAA. Somatic embryogenesis was induced using different explants, including immature seeds as well as leaves and roots from shoot cultures. Following auxin treatment with either 1.0 mg L −1 2,4-D (immature seeds and leaves) or 0.1 mg L −1 NAA (roots), explants produced embryogenic calli which upon transfer to plant growth regulator-free medium allowed embryo conversion into plantlets. The best embryogenic response (82%) was obtained in calli derived from leaves cultured with their abaxial surface in contact with medium, whereas the highest embryo conversion rate (68%) was recorded for calli induced on immature seed explants. Histological analysis in all explant types revealed development of proembryogenic cell complexes at callus periphery, giving rise to somatic embryos. The presence of embryos at various stages of development indicated asynchronous somatic embryogenesis in G . utriculosa . Derooted embryo-derived plantlets placed on medium with 0.2 mg L −1 BA multiplied further as shoot cultures.

The best duration of cold treatment for germination of Saxifraga aizoides and S. paniculata was investigated comparing seeds collected from Alpine and Apennine populations. For S. aizoides seeds of both provenances, the highest germination was obtained after 12 weeks of cold treatment, and Apennine seeds showed the best performance. For S. paniculata , the effect of cold treatment differed greatly in relation to provenance: Alpine seeds showed theirs highest germination after 20 weeks, while Apennine seeds showed their best, albeit low, germination percentage after 8–16 weeks. For both species, a maximum germination plateau was reached 20 days after sowing. Thus, for ex situ conservation and restoration projects in the Apennines, we suppose that S. aizoides seeds should be collected from the location closest to the restoration site and S. paniculata seeds from the Alps. These results also suggest that specific responses to cold treatment exist within the genus Saxifraga in relation to provenance.

Ribosome-inactivating proteins (RIPs), a type of defensive chemicals, play key roles in host defense mechanisms against foreign pathogenic invaders and plant stress resistance. Alpha-momorcharin, a member of typical type I RIPs isolated from Momordica charantia , showed strongly antifungal, antitumor, and antibacterial activities both in vitro and in vivo . In this study, the expression level of alpha-momorcharin gene in response to environmental stress and phytohormones were examined through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting analyses. Results showed that α -momorcharin gene expression was up-regulated in response to salinity and osmotic stresses. Phytohormones, including abscisic acid and gibberellic acid, also enhanced α -momorcharin expression, while the expression level was not affected by salicylic acid. It’s speculated that the α -momorcharin gene might be functional in plant basal or defense resistance against environmental stresses, and it also involved in hormonal signaling pathways. This research might be exploited in further genetic engineering programs and plant protection.

Living roots grow in soil, which is a heterogeneous environment containing a wide variety of physical barriers. Roots must avoid these barriers to grow: first, they adopt a characteristic S-shape that can be described by the angle between the root tip and the barrier (i.e., the tip-to-barrier angle); then, they move parallel to the barrier by keeping the sensitive tip in contact with the barrier until it has been circumvented. We investigated this avoidance response in the primary roots of maize ( Zea mays ) by considering flat barriers oriented at 45, 60 and 90 degrees with respect to the gravity vector. We measured the root tip orientation during growth by using time-lapse imaging and specially developed tip-tracking software (9 trials for each value of the barrier orientation). Remarkably, we found that the S-shapes formed by the roots were characterized by the same tip-to-barrier angle regardless of the barrier orientation: namely, 21.96 ± 2.97, 21.48 ± 4.75 and 20.81 ± 9.39 degrees for barriers oriented at 45, 60 and 90 degrees, respectively. We also considered the root growth after bypassing the barrier; for the barrier at 90 degrees, we observed a gravitropic recovery. Furthermore, we used a mathematical model to quantify the characteristic time of S-shape formation (95 min on average) and gravitropic recovery (approximately 42 min); the obtained values are consistent with those of previous studies. Our results suggest that the avoidance response develops with respect to a reference frame associated with the barrier. From a biological viewpoint, the reason the root adopts the specifically observed tip-to-barrier angle is unclear, but we speculate that maize root optimizes energy expenditure during the penetration of a medium.

The earthworm genus Satchellius Gates, 1975 is reviewed. On the basis of the presence of posterolateral calciferous sacs in segment 10 the species Dendrobaena alvaradoi Moreno, Jesus & Diaz Cosin, 1982 and Dendrobaena madeirensis (Michaelsen, 1891) are transferred to the genus Satchellius as comb. novae. The specimens reported as Allolobophora madeirensis by Zicsi (1969) from Madeira Island (Portugal) are described as Zophoscolex zicsianus sp. n.

Serum of mud crab, Scylla serrata had antibacterial activity against certain host specific bacteria as well as other known crustacean pathogenic bacteria. The respective antibacterial molecule was isolated by ion-exchange chromatographic method using diethylaminoethyl-cellulose matrix and eluted with gradient increase of sodium chloride (0-0.5 M). There were three major peaks on the elution profile and fractions from second major peak exhibit differential antibacterial response against Bacillus flexus N3, Escherichia coli , Vibrio harveyi and Vibrio vulnificus . The electrophoretic analyses of this fraction resulted in single protein band and the native molecular weight of this protein was found to be approximately 305 kDa. The bathocuproine sulfonic acid staining (a specific stain for copper), copper/protein ratio (0.129%) reveals this isolated antibacterial protein to be hemocyanin. This antibacterial hemocyanin is also capable of binding to cations, namely Ca 2+ , K + , Mg 2+ , Mn 2+ and Zn 2+ apart from carrying Fe 2+ ions. This native antibacterial hemocyanin was found to contain three or four subunits with possible molecular size of 70-98 kDa under reducing conditions. These findings indicate that the respiratory protein hemocyanin in the presence of cations functions as humoral immune molecule and the detailed investigation would reveal additional immune functional characteristics of this respiratory protein as well as subunits-specific functions of this antibacterial hemocyanin.

The two-year dynamics of soil collembolan communities were studied at five sites along the mesoclimatic gradient in a deep karst valley, from the karst plateau to the bottom of the deep karst gorge in the Zádiel Valley, Slovak Karst, Slovakia. Within the local gradient the annual soil temperature means ranged from +8.88 ± 6.72°C on the plateau (pasture) to +5.16 ± 6.24°C at the bottom of the gorge (maple-hornbeam wood). Repeated ANOVA confirmed a significant effect of site, year, month and combined factors (year × month, year × month × site) on Collembola abundance ( P < 0.0001). The marked spring minimum in abundance dynamics and species richness of communities recorded in the gorge were not observed at the thermophilous sites. The collembolan abundance means at both sites in the gorge were extremely low and significantly different ( P < 0.001) from those of the rest of the study period, delayed into the early summer as a direct effect of mesoclimatic inversion within the karst valley. During the drier year a more marked gradient was observed in the deep karst valley, leading to greater changes in the dynamics of dominant species Isotomiella minor and Parisotoma notabilis .

Bromeliads accumulate water in the leaf axils (forming phytotelmata), which may then form a habitat for many animal species of both invertebrates and vertebrates. In temperate regions, bromeliads are commonly displayed in palm houses and distributed by florist wholesalers. Taking into account the fact that global plant trade can be a source of spreading alien species, the phytotelmata that had formed within the bromeliads were examined here in order to study the protists and invertebrates associated with this habitat. In the examined Bromeliaceae micro-reservoirs representatives of testate amoebae (Euglyphida and Arcellinida), Gastrotricha, Nematoda, Rotifera, Polychaeta, Cladocera, Copepoda and mites (Oribatida, Mesostigmata and Heterostigmatina) were found. Additionally, unidentified Nemertea, Turbellaria, Insecta larvae and single representative of Mollusca were also found. Thirty taxa are recorded for the first time from bromeliads micro-reservoirs. Among the identified taxa, the gastrotrich Chaetonotus (Hystricochaetonotus ) furcatus Kisielewski, 1991 was found, which was previously known only from bromeliads in the natural environment in South America. Nematodes belonging to Clarkus spp. were found in the Bromeliaceae reservoirs for the first time. Additionally, the tropical mite Phthiracarus (Phthiracarus ) phoxos Niedbała, 2004, previously known from Cuba in the natural environment and other greenhouses in Poland, was also found in the studied phytotelmata. Steneotarsonemus ananas (Tryon, 1898) sensu Beer, 1954 was recorded from Poland for the first time. Some of the mite specimens, e.g., Mesostigmata and Heterostigmatina, had most likely accidentally been caught in the phytotelmatum, which acted like a trap; but others, especially freshwater fauna representatives can be a constant element of the phytotelmata merocenoses. Our study provides comprehensive analysis of fauna living in these exceptional micro-habitats, and it underlines the possibility of passive invertebrate dispersal within bromeliad micro-reservoirs and between different localities, e.g., florist shops and markets as well as greenhouses.

In years 2010–2013, we collected mosquito larvae and adults at selected locations in Eastern Slovakia. We have identified 22 mosquito species belonging to 6 genera: Aedes (4 species), Anopheles (4 species), Coquillettidia (1 species), Culiseta (1 species), Culex (3 species) and Ochlerotatus (11 species), with eudominant representation of Culex pipiens / Cx. torrentium (42.36%), Aedes vexans (36.65%) and Ochlerotatus sticticus (12.15%). As for larvae, we collected 16 species belonging to 5 genera: Aedes (4 species), Anopheles (1 species), Culex (2 species), Culiseta (1 species) and Ochlerotatus (8 species). Out of the total number of 29,155 specimens of identified larvae, eudominant species were again Cx. pipiens (33.80%), Ae. vexans (19.27%) and Oc. cantans (15.99%). Seasonal changes in the population dynamics of individual species of larvae, as well as adults, reflect different climatic conditions of local sites.

In order to determine effects of different types of supplementary feed on the histology of liver and intestine of carp reared in a semi-intensive system, a six-month field experiment was carried out. Carp yearlings were fed different supplemental feeds: cereals (CF), pelleted (PF) and extruded (EF) compound feeds. Hepatocytes histomorphometry revealed larger nuclear profile area of EF-fed fish compared to other two groups, while cytoplasmic profile area of the hepatocytes decreased in following sequence: CF > EF > PF. Lower nucleus/cytoplasm ratio was found in CF-fed fish compared to EF-and PF-fed fish. Striking, season dependant finding, was that regardless of the type of supplementary feed the number of lipid droplets in hepatocytes was highest in July and August, lipid depletion occurred in September and October, while glycogen was present in hepatocytes throughout the study. No size difference between groups was observed for enterocytes height, absorptive surface height and number of goblet cells in tunica mucosa , but a strong relationship between length of intestinal folds and fish weight was found. Histological analysis revealed a difference among three types of supplemental feed used in this field study: extruded or steam-pelleted feed induced better nutritional status of common carp in comparison to cereals. At the same time, histomorphometry indicated some differences between EF-and PF-fed fish.

The concentrations of six elements (Ca, Mg, Zn, Cu, Pb and Cd) in the eggshells of breeding red-crowned cranes ( Grus japonensis ) in China’s Zhalong Wetland were examined. Two macronutrients, namely, Ca and Mg, were predominant in the eggshells, with corresponding concentrations varying from 127 g kg –1 to 323 gkg –1 and 0.33 g kg –1 to 5.51 gkg –1 (dry weight, dw), respectively. The Cu and Zn contents were in the range of below the detection limit (ND) to 8.05 mg kg −1 and ND to 511 mg kg −1 dw, respectively. The maximal Pb and Cd concentrations in the eggshells of the red-crowned crane exceeded a level considered to be potentially toxic in birds (i.e., 1.7 mg kg −1 for Pb and 0.34–0.91 mg kg −1 dw for Cd), ranging from ND–3.20 mg kg −1 dw for Pb and ND–1.20 mg kg −1 dw for Cd. The principal components analysis results revealed the complex chemical associations between two essential (Cu and Zn) and nonessential (Pb and Cd) elements in the eggshells of G.japonensis in China. Increased Pb and Cd levels in the bodies and eggshells of the red-crowned cranes suggest this species in their colonies are exposed to these contaminants, although which had not induced to obvious eggshell thinning in the examined species.