Volume 71, Issue 6

DNA was extracted using a Silica Bead Extraction kit from bone samples taken from a Slavonic-Avar individual found at the archaeological site of Cífer-Pác. The analysed skeletal remains from the grave number 62/79 belong to a young adult male (20–30 years at death) and are dated to the 8 th –9 th century anno Domini. The isolated ancient DNA (aDNA) was amplified by a targeted PCR with a primer pair designed to recognize the Mycobacterium tuberculosis complex insertion sequence IS6110. The aim of this molecular approach was to test and optimize a methodology for aDNA M. tuberculosis complex extraction from bone samples with osteological evidence of tuberculosis. Despite of the currently biased authenticity of the mentioned fragment, in this case study we prove that macroscopic evidence for tuberculosis additionally supported by a positive result of molecular testing can be considered authentic enough to be the proof of a tuberculosis infection caused by MTBC ( Mycobacterium tuberculosis complex) bacteria when additional skeletal trauma and changes potentially caused by MOTT (mycobacteria other than tuberculosis) bacteria can be excluded. Positivity was confirmed in all of the three samples (thoracic vertebrae, lumbar vertebrae and right femur). Our results confirmed the diagnosis of tuberculosis of the spine and right hip joint. This is the first molecular evidence for the occurrence of tuberculosis on the territory of Slovakia. Through this bio-molecular approach we wish to provide a basis for aDNA examinations on other skeletal collections and provide epidemiological data concerning historical populations living on the territory of Slovakia.

Glaciers constitute freshwater reservoirs. They are the main contributors to sea level rise, play an important role in global carbon cycle and are the source of water for people. Despite extreme conditions, they are also a habitat for a number of cryophilic organisms. To fill the gap in the knowledge on the biota on glaciers in Caucasus, the aim of the study was to: (i) isolate and enumerate culturable heterotrophs; (ii) determine the number of coliforms and enterococci bacteria; and (iii) analyze total microbial 16S rRNA gene in cryoconite sediments, ice and gravel. The material was collected from two glaciers in Caucasus (Georgia, Svaneti region). Bacteria Aeromonas sp. and Pseudomonas sp. were found. The total number of culturable heterotrophic bacteria ranged from 0.2 × 10 1 colony forming units per mL in ice and gravel of the Chalaadi Glacier to 3.9 x 10 3 colony forming units per mL in a cryoconite sediments of the Adishi Glacier. The average number of 16S rRNA gene copies ranged from 5.3 × 10 5 /g to 5.3 × 10 6 /g in ice and gravel from Chalaadi Glacier and from 3.1 × 10 5 /g to 1.5 × 10 7 /g in a cryoconite of the Adishi Glacier. The 16S rRNA gene of five Pseudomonas sp. isolates was sequenced. Moreover, in the analyzed cryoconite material, a rare collembolan Gnathisotoma sp. was found, which constitutes the first record of springtails in cryoconite holes in the Northern hemisphere.

To enhance the expression level of recombinant feruloyl esterase in Pichia pastoris , the Aspergillus niger feruloyl esterase ( AnFaeA ) gene was optimized based on the codon usage bias of P. pastoris and synthesized. The synthetic AnFaeA-syn was inserted into the pPICZαA and then integrated into P. pastoris X33. SDS-PAGE analysis of recombinant feruloyl esterase displayed a single protein band with an apparent molecular mass of 40.0 kDa. Western blot analysis confirmed that the expressed protein was the target protein. Furthermore, we showed that codon optimization increased enzyme yield for about 6.4-fold (35.12 U/mL vs 5.51 U/mL) in shake flask cultivation, compared to the non-codon-optimized control. Taken together, this study showed that the yield of feruloyl esterase could be increased by codon optimization.

In this study we focused on the comparison of the physiological responses to excess nitrogen (N) comparing Cladonia arbuscula subsp. mitis and Cladonia furcata . Both lichens were exposed to increased amounts of nitrogen (NH 4 NO 3 ) for five weeks. Different concentrations of nitrogen were applied by spraying the lichen thalli during this period. After the treatment, the physiological parameters of the lichen, like chlorophyll a fluorescence, chlorophyll b integrity, content of soluble proteins, ergosterol, thiobarbituric acid reactive substances (TBARS) and content of secondary metabolites were measured. We found out that lichens responded differently to nitrogen excess concerning photobiont and mycobiont. The mycobiont of C. arbuscula subsp. mitis seems to be more sensitive to the nitrogen excess than C. furcata based on decreased ergosterol content. We concluded that the mycobiont is more sensitive to nitrogen excess than the photobiont in case of the tested lichen species in a middle term experiment.

The species-rich grass genus Sesleria is one of the most intricate genera of the European flora. Almost all species of the genus Sesleria are polyploid; only two species from the Alps, previously segregated as separate genera Psilathera and Sesleriella , are diploid. In addition, there is controversy about two diploid counts of S. comosa from the Bulgarian Rila and Pirin mountain ranges. These are the single reports of diploidy in Sesleria s. str., which would have significant impact on the future phylogenetic reconstruction of the genus’ evolutionary history. In order to confirm or reject these reports, we determined the relative DNA-content for 67 individuals of S. comosa from the two Bulgarian localities, from which diploid cytotypes have been reported. All analyzed individuals were clearly separated into two ploidy levels: DNA-tetraploids and DNA-octoploids. These results are in accordance with our previous comprehensive study on genome sizes in the genus, but are in stark contrast to the previous reports. Therefore, we suggest that the diploid counts rather refer to co-occurring, morphologically somewhat similar diploid Alopecurus gerardii , which has probably overgrown and outcompeted the slower growing and more susceptible S. comosa during cultivation before root tips were fixed for chromosome counting.

RCD1 protein confers multiple stress tolerance to plant, while playing in it developmental roles as well. After an extensive characterization of its gene in A. thaliana , role of its orthologues under varying stresses have also been characterized in rice, wheat and tomato plants. RCD1 of A. thaliana with its rice homologue has been found consensually involved in providing plant tolerance against oxidative and osmotic stresses; however, its stress-tolerance roles under other environmental stresses seem rather contrasting. This is also the case with the expression profile, primary protein structure and binding affinities of these two homologues. Such differences in behaviours of the RCD1 homologues raises need to study their evolutionary status, in an effort to infer relative conservation of their functions. To elucidate this pattern, available RCD1 homologues have been subjected to Maximum Parsimony and Bayesian Inference-based analysis in this research. Conservation status of the SRO1/RCD1 type-specific WWE domain has also been explored using the available sequences in online databases along with the WWE-encoding sequences obtained in sugarcane, wheat and rice using single set of primers designed on conserved flanking regions. Results suggest a high conservation of these homologues within plant families, suggesting family-specific conservation of their functionalities in demonstrating stress responses.

Nucleolus was regarded as a mere ribosome-producing factory for many decades however, in the past two decades; this has come to the light that it is a plurifunctional nuclear domain which plays vital role in many cellular processes. Despite many advances, much remains to be revealed about the nucleolus association with cellular processes. While exploring the chromosomal diversity in the high altitude plants of the Indian cold desert region in North West Himalayas we came across 140 wild plants, out of these, pollen mother cell (PMC) in 31 plants showed the occurrence of additional nucleoli. Coincidently, all these plants also depicted the phenomenon of cytomixis i.e. inter-PMC chromatin material extrusion/transfer among/between adjacent meiocytes. Interestingly, it was noticed that nucleolus accompanied the chromatin material during process of cytomixis. During, and after the migration of nucleolus, change in its shape, size and number has also been discussed. So the current paper here in correlated the inter-PMC migration of nucleolus to the occurrence of cytomixis.

MicroRNAs (miRNAs) play key roles in plant responses to biotic and abiotic stresses by modulating their own expression and a wide array of target mRNAs. Reverse transcription quantitative real-time PCR is a sensitive and widely used method to study miRNA expression profile. Accurate analysis and interpretation of results require the selection of an appropriate reference gene. Reference genes selected should have a constant expression level under different stress conditions. Six reference candidates, including two miRNAs (miRNA 156 and miRNA 172), an rRNA (5S rRNA), a snRNA (U6) and two protein coding genes (actin and protein phosphatase 2A), were selected for normalization of miRNA expression in Lablab purpureus . The expression stability of candidate reference genes was investigated in ten samples and analysed using NormFinder, BestKeeper and hkgFinder softwares. The analyses suggested that miRNA 156 is the appropriate reference gene, as it had better expression stability than protein-coding genes, and other non-coding RNAs.

For a long time ecologists have questioned on the variations of biodiversity across the latitudinal gradient. Recently, it has emerged that the changes in β -diversity are caused simply by changes in the sizes of species pools. In this study, the species pool size and the fractal nature of ecosystems was combined to clarify some general patterns of this gradient. Considering temperature, humidity and NPP as the main variables of an ecosystem niche and as the axis of the polygon in the Cartesian plane, it is possible to build fractal hypervolumes, whose fractal dimension rises up to three moving towards the equator. It follows that the best figure that graphically synthesizes the evolutionary forces that fit this ecosystem hypervolume is the fractal cauliflower.

A new species of oribatid mites of the subgenus Neoribates ( Neoribates ) (Oribatida: Parakalummidae), Neoribates ( Neoribates ) peruensis Ermilov sp. n., is described from soil and leaf litter in forests of Peru and Ecuador. This species differs from N . ( Neoribates ) fulvus Sellnick, 1923 by the smaller body size and the presence of prodorsal longitudinal ridge and straight anterior margin of notogaster. An identification key to the Neotropical species of Neoribates ( Neoribates ) is presented.

Habitat specialists and generalists are known to differ in their width of environmental tolerance and their representation can vary along with the ecological contrast of habitats. In this study we explore factors shaping patterns of species richness and abundance of aquatic macroinvertebrate habitat specialists and generalists at isolated spring fens, separately for spring patch and spring brook mesohabitats at each site. We also examined habitat contrast of these unique island-like communities by the comparison of spring fen specialists and habitat generalists shared between the two spring fen mesohabitats and the nearest stream to each of 13 selects fen sites. Aquatic macroinvertebrates (Clitellata, Ephemeroptera, Plecoptera, Trichoptera, and Diptera) were investigated at 62 isolated spring fens, with 357 taxa identified in more than 172,000 individuals collected. We found that specialists experienced a stronger relation to local environmental conditions (i.e. the amount of dissolved oxygen and water conductivity) at both spring mesohabitats than generalists, primarily responding to fen habitat size. In contrast, responses of species abundances at spring patches and spring brooks differed as the abundances were controlled by the amount of oxygen in spring patches and by habitat size in spring brooks. Based on Trichoptera and Diptera assemblages we found a similar contrast between both spring fen sites and nearby streams. Our results suggest a higher resilience of specialist populations in well oxygenated sites and their competitive advantage over generalists at these sites, which stresses the importance to prevent any significant decrease of oxygenation (e.g., by eutrophication or drainage), especially in spring patches.

The first overview on the Heteroptera of peat bogs of the Belarusian Lakeland is presented. Five natural oligotrophic peat bogs were studied in 2012–2014 by entomological sweep-net. A total of almost 2,612 true bugs were sampled, representing 83 species of 13 families. Ten species (12.11–36.72% of the individuals) were tyrphobiontic or tyrphophilous. The five most abundant species represented 71.39–82.30% of the individuals. The most common species were Stictopleurus crassicornis (14.56–59.63%) and Stephanitis oberti (10.25–36.21%). The number of individuals of adult Heteroptera in peat bogs was low in spring and high in July-August. The natural peat bog offered favorable conditions to phytophagous (72.29%) polyphagous (60.00%) true bugs species. The most preferred host plants were ericales shrubs.

Monoacylglycerol O-acyltransferase 2 (MOGAT2) plays critical roles in lipid homeostasis. We reported a cDNA designed BmMOGAT2 encoding an MOGAT2 homologue cloned from the fat body of the silkworm Bombyx mori , by using conserved domain homology search method. The resultant BmMOGAT2 was translated to a protein encoding 352 amino acids with a theoretical isoelectric point of 9.04 and a molecular weight of 39,944.48 Da. Homology analysis revealed that BmMOGAT2 exhibits higher similarity on the amino acid level to those of other species already reported; 48% identity with Homo sapiens , 46% with Mus musculus , 50% with Danio rerio , and 42% with Drosophila melanogaster . The expression of BmMOGAT2 was detected in all tissues tested with 2-fold higher expression in the post-silk gland, as compared to others, and stronger expression of the larval fat body at 1 st instar, as compared with other stages. The BmMOGAT2 is a predicted endoplasmic reticulum (ER) transmembrane protein with two ER transmembrane domains; BmMOGAT2 gene expression increases in response to ER stress-inducible drugs. To our knowledge, this is the first report of Bombyx mori MOGAT2 cDNA, BmMOGAT2 , and its associated molecular characterization.

The aim of this study was to identify the lingual surface and posterior lingual glands in the red kangaroo (Macropus rufus) using light and scanning electron microscopy (SEM). Haematoxylin and eosin (H&E), Masson-Goldner trichrome and Azan trichrome stainings were used for the histological examinations of both the papillae and concomitant glands, located beneath the body of the papillae. The samples were stained also with Alcian blue pH 2.5 and periodic acid-Schiff for histochemical observations. A scanning electron microscope (Tesla BS - 300) was used to examine the lingual surface. Small, giant and elongated filiform mechanical papillae and fungiform, vallate and foliate gustatory papillae were recognized. No typical conical or lenticular papillae were found on the lingual surface. The distribution of each group of papillae varied on the whole surface of the tongue. The majority of the fungiform papillae resembled “bud-like” shapes, and rotund taste buds were present within the fungiform papillae epithelium. The SEM study showed that the apical part of the vallate papilla was oval with an irregular surface. Additionally, the internal wall of this papilla had many intraepithelial cylindrical or rod-shaped taste buds. The papillary leaves did not have any typical appearance, however, the histological study showed the presence of intra-epithelial taste buds in the wall of the foliate papillae area. Furthermore, two types of posterior mixed mucoserous and serous lingual glands were observed. In conclusion, the morphology of the lingual surface as well as the structure of the lingual glands facilitates the consumption of particular foods in the red kangaroo. Additionally, there are typical microstructural features of the red kangaroo tongue, which are different than in other marsupials.

Of all alien parrots, the rose-ringed parakeet Psittacula krameri is the most successful invasive species, with established populations in at least 35 countries. This paper reports a study, conducted from March to August 2015, on the population size in Lisbon (Portugal) using roost counts. We found only one important roost for this parrot in Campo Grande, where a maximum of 644 individuals were counted in July. The recorded information suggests that this species has increased considerably since the last counts done. The present status of this species in the city may lead to a further expansion into new nearby areas.

Introduced fish represent a major threat for aquatic ecosystems and species and their impact on semiaquatic vertebrates, such as amphibians, has been extensively studied. On the other hand, little is known about their impact on semiaquatic mammals, such as water shrews ( Neomys spp., Soricidae), although small mammals have been reported in their diet. The present note reports on a case of predation on an adult Eurasian water shrew Neomys fodiens by a large sized brown trout Salmo trutta belonging to a population introduced in a small pastureland pond in an Italian mountain area. The authors considered that shrews, including relatively large species such as Neomys spp., can be susceptible to predation by introduced fish and they use this finding to discuss about direct and indirect impacts of introduced fish on the populations of shrews inhabiting originally fishless habitats such as headwaters.