Isolation of a newLegionella species from thermally altered waters

Legionella-like bacteria were isolated from thermally altered water in Pennsylvania and Minnesota. These organisms were isolated from tissues of infected guinea pigs after intraperitoneal inoculation of water concentrates. While the cultural and morphological characteristics indicated the isolates wereLegionella, they did not react with antibodies prepared against known species of the genus. Antisera prepared against one of the isolates reacted maximally with the other ten isolates. Fluorescent antibody analysis of water concentrates from geographically disparate sites indicated that the environmental distribution is broad and that concentrations of the Oak Ridge strain ofLegionella were similar to those of serogroups 1, 2, 3, and 4 ofLegionella pneumophila.     

Antimicrobial activity of some Indian plants

Antimicrobial activity of 105 Indian plant species was tested. Among them, 30 showed antibacterial activity; 20 of these exhibited antifungal action as well. Seeds ofCarum copticum, stem ofPinus longifolia, roots ofPlumbago zeylanica andSaussurea lappa, and rhizome ofAlpinia officinarum have considerable antifungal activity, especially against pathogenic fungi. Antibiotic activity against a wide variety of microorganisms—pathogenic and nonpathogenic gram-positive and gram-negative bacteria, yeast, and fungi—was also noted with leaves ofLawsonia inermis, roots ofPlumbago zeylanica, and fruits ofTamarindus indica,Terminalia belerica, andEmblica officinalis.     

Morphological differentiation of flower buds in Apple-Trees

The paper deals with the study of morphological differentiation of flower buds in 10 varieties of apple-trees in relation to the growth of summer shoots and the earliness of varieties. The data obtained indicate that in the warmer regions of Slovakia the critical season of transformation of flower buds from vegetative to generative forms takes place, on the average, between July 20 and 29, and, at the same time, the variety Yellow Transparent deviates from the average four week-earlier. According to the author we can speak neither of a positive relation between the onset of morphological differentiation and the active growth of summer shoots nor of the relation between the earliness of the variety in maturing and the onset of differentiation. The morphological differentiation may be classified into five phases: 1^st phase — the formation of the growing point primordium, 2^nd phase — the formation of the cup on sepal primordia, 3^rd phase — the formation of petal primordium, 4^th phase — the formation of carpel and stamen primordia, 6^th phase — completion of the differentiation through substantial enlargement of flower organs before the onset of winter.     

Diversity and variability of methanogens during the shift from mesophilic to thermohilic conditions while biogas production

Anaerobic digestion (AD) is the most popular path of organic waste disposal. It is often used in wastewater treatment plants for excessive sludge removal. Methanogenic fermentation had usually been performed under mesophilic conditions, but in the past few years the thermophilic processes have become more popular due to economics and sludge sanitation. Methanogens, the group of microorganisms responsible for methane production, are thought to be sensitive to temperature change and it has already been proven that the communities performing methanogenesis under mesophilic and thermophilic conditions differ. But in most cases the research performed on methanogen diversity and changeability was undertaken in two separate anaerobic chambers for meso- and thermophilic conditions. It is also known that there is a group of microorganisms performing AD which are insensitive to temperature. Also the linkage between digester performance and its microbial content and community changeability is still not fully understood. That is why in this experiment we analyzed the bacterial community performing methanogenesis in a pilot scale anaerobic chamber during the shift from mesophilic to thermophilic conditions to point at the group of temperature tolerant microorganisms and their performance. The research was performed with PCR–DGGE (polymerase chain reaction–denaturing gradient gel electrophoresis). It occurred that the community biodiversity decreased together with a temperature increase. The changes were coherent for both the total bacteria community and methanogens. These bacterial shifts were also convergent with biogas production—it decreased in the beginning of the thermophilic phase with the bacterial biodiversity decrease and increased when the community seemed to be restored. DGGE results suggest that among a wide variety of microorganisms involved in AD there is a GC-rich group relatively insensitive towards temperature change, able to adapt quickly to shifts in temperature and perform AD effectively. The studies of this microbial group could be a step forward in developing more efficient anaerobic digestion technology.     

The application of DNA sequence data for the identification of benthic nematodes from the North Sea

Nematodes or roundworms represent one of the most diverse and dominant taxon in marine benthic habitats. Whereas a morphological identification of many species is challenging, the application of molecular markers represents a promising approach for species discrimination and identification. In this study, we used an integrative taxonomic approach, combining both molecular and morphological methods, to characterize nematodes of distinct sex and ontogenetic stages from three sampling sites of the North Sea. Morphospecies were discriminated after first visual determination, followed by a molecular analysis of the nuclear 28S rDNA: D2–D3 marker. By linking each sequence to a morphological voucher, discordant morphological identification was subjected to a so-called reverse taxonomic approach. Molecular operational taxonomic units (MOTUs) and morphospecies were compared for all of the three sampling sites to assess concordance of methodology. In total, 32 MOTUs and 26 morphospecies were assigned, of which 12 taxa were identified as described species. Both approaches showed high concordance in taxon assignment (84.4 %) except for a cluster comprising various Sabatieria species. Our study revealed the high potential of the analyzed fragment as a useful molecular marker for the identification of the North Sea nematodes and highlighted the applicability of this combined taxonomic approach in general.     

Biosynthesis,characterization and antimicrobial activity of silver nanoparticles by Streptomyces sp. SS2

In the present study the microbial biosynthesis of silver nanoparticles (AgNPs) by secondary metabolites of Streptomyces sp. SS2 in an eco-friendly approach has been reported. The Streptomyces sp. SS2 was isolated from the soil sediment of Similipal Biosphere Reserve. The identification of this strain was determined by phenotypical characteristics (morphological and biochemical) and molecular characterization method using 16 s rDNA sequencing. The morphological study was also done by high-resolution scanning electron microscopy. The preliminary characterization of biosynthesized silver nanoparticle was carried out using UV–Vis spectrum analysis, which showed an absorption peak at 420 nm corresponding to plasmon absorption of silver. The average size and charge (zeta potential) of the particles were found to be 67.95 ± 18.52 nm and ?17.7 ± 5.30 mV, respectively. The functional groups were identified by FTIR studies and their morphology (round and spherical shape) was determined by scanning electron microscopy. The synthesized AgNPs exhibited excellent antibacterial activity against Escherichia coli (MTCC 1089), Bacillus subtilis (MTCC 7164), Staphylococcus epidermis (MTCC 3615), Vibrio cholerae (MTCC 3904) and Staphylococcus aureus (MTCC 1144). These biotechnological approaches of synthesis of nanoparticles can direct a new path in biomaterial sciences and enrich biomedical applications.     

Combining immunofluorescence with in situ proximity ligation assay: a novel imaging approach to monitor protein–protein interactions in relation to subcellular localization

The in situ Proximity Ligation Assay (PLA) is suited for visualizing protein–protein interactions and post-translational protein modifications in both tissue sections and in vitro cell cultures. Accurate identification and quantification of protein–protein interactions are critical for in vitro cell analysis, especially when studying the dynamic involvement of proteins in various processes, including cell proliferation, differentiation, and apoptosis. Here, we monitored the interactions between protein kinase-Cζ (PKCζ) and Bcl10 protein in untreated and etoposide (VP-16)-treated C4-I cells by means of a new combined morphological approach and validated it by taking stock of our previous proteomic and biochemical work (Chiarini et al. in J Proteome Res 11:3996–4012, 2012). We first analyzed the colocalization of PKCζ and Bcl10 proteins through classical immunofluorescent colocalization analysis. On the basis of these results, we developed a novel imaging approach combining immunofluorescence (IF) techniques with in situ PLA to identify the PKCζ·Bcl10 complexes at the level of a specific subcellular compartment, i.e., the nuclear envelope (NE). By this means, we could show that the amount of PKCζ·Bcl10 complexes localized at the NE of C4-I cells during proliferation or after treatment with VP-16 closely corresponded to our previous purely biochemical results. Hence, the present findings demonstrate that the combination of in situ PLA with classical IF detection is a novel powerful analytical tool allowing to morphologically demonstrate new specific protein–protein interactions at level of subcellular organelles, the complexes functions of which can next be clarified through proteomic/biochemical approaches.     

Coupling scanning electron microscopy with DNA bar coding: a novel approach for thrips identification

The small size and cryptic nature of thrips pests often make their monitoring and the identification process difficult. With the aim to obtain accurate identification of various pest species in the order Thysanoptera, a pilot study was conducted on an invasive thrips species Scirtothrips dorsalis Hood in Florida. The specific objective of this pilot study was to assess if the thrips specimens processed to be observed under scanning electron microscopy (SEM) can be further utilized for DNA based assays. Larvae and adults of S. dorsalis were subjected to traditional morphological identification using high resolution SEM prior to their DNA extraction. Sequence results of both mtCO1 and ITS rDNA of individual larva and adult S. dorsalis were found to be in agreement with the taxonomic identification conducted using SEM and each result confirmed the other technique. Our results suggest that steps involved during specimen preparation and observation under SEM does not impact DNA analysis of the sample. The two techniques together could be used for the correct identification of various thrips species using the same specimen. The proof of concept was further tested on three other important thrips species Frankliniella occidentalis (Pergande), F. schultzei (Trybom) and Thrips palmi Karny, which confirmed the robustness of the technique. The technique has broader significance for different areas under entomological sciences especially medical, forensic and taxonomic studies where the samples put under a high beam of ions for investigation are often required for DNA analysis.     

Is the insular endemic Psidium socorrense (Myrtaceae) at risk of extinction through hybridization?

Natural hybridization between an insular endemic species and a widely distributed congener may endanger the endemic through genetic assimilation or outbreeding depression. Furthermore, hybrids can exhibit complex morphological variation, causing taxonomic problems in the identification of the involved taxa. In this work, we used a combination of leaf morphological and molecular markers (RAPD) to establish the differentiation between Psidium sp. aff. sartorianum and the insular endemic P. socorrense. It was also determined if hybridization between these taxa occurs in the southern slope of Isla Socorro, Mexico. Plant collection was carried along an altitudinal gradient (100–800 m). We collected eight populations separated 100 m a.s.l. apart from each other; 25 individuals were collected per population. Psidium socorrense and P. sp. aff. sartorianum differed significantly in all but two morphological characters measured. Also, a high number of diagnostic RAPD markers were found for each taxon. These results suggest that two Psidium species occur at Isla Socorro. Furthermore, both morphological and RAPD markers revealed a hybrid zone located in the southern slope of Isla Socorro (400–700 m a.s.l.) with an asymmetrical pattern of gene flow towards P. socorrense. We suggest that the disturbance caused by the sheep population in the mixed stand favors the establishment of hybrids. We further discuss whether hybridization represents a threat to the insular endemic P. socorrense.     

Magainin 2 Induces Bacterial Cell Death Showing Apoptotic Properties

Magainin 2 is pore-forming antimicrobial peptide on lipid matrix of bacterial membrane, secreted from the skin of the African clawed frog Xenopus laevis. The aim of this study was to investigate a new concept for antibacterial mechanisms called bacterial apoptosis-like cell death. We examined the morphological changes induced by magainin 2 in Escherichia coli, regarding apoptosis. Specifically, phosphatidylserine externalization from the inner to outer membrane surface was detected by Annexin V staining, and DNA fragmentation and chromatin condensation was detected by TUNEL and DAPI assay. We also found much mechanistic evidence to support the hypothesis that magainin 2 induces bacterial apoptosis-like death—including disturbance of membrane detected by DiBAC₄(3), caspase activation observed by FITC-VAD-FMK staining, and analyzing the role of RecA in bacterial apoptosis-like death through the RecA expression assay by Western blot—in E. Coli when treated with magainin 2. On the basis of these results, magainin 2 exerts antibacterial activity with a new mechanism which is bacterial apoptosis-like death. Searching antimicrobial agents with novel mechanisms of action can be an effective strategy to coping with the emergence of new resistance mechanisms. Magainin 2 deserves further research as a potential antimicrobial therapeutic agent.     

A study of Schizoblastosporion starkeyi-henricii isolates from northern and southern hemispheres of the Earth

A species Schizoblastosporion starkeyi-henricii is common in peats and peaty soils. Fifteen strains of this species isolated from Europe and East Falkland Island were found homogenous in cultural, morphological and physiological characters, with the exception of one strain that was sensitive to elevated osmotic pressure. Analyses of the D1/D2 region of the 26S rDNA and internal transcribed spacers (ITS) confirm that these strains and also Asian isolates are conspecific. Our results show that ecological factors but not geographic distances determine the global distribution of yeast fungi.     

O-Succinyl-l-homoserine-based C4-chemical production: succinic acid,homoserine lactone, γ-butyrolactone, γ-butyrolactone derivatives,and 1,4-butanediol

There has been a significant global interest to produce bulk chemicals from renewable resources using engineered microorganisms. Large research programs have been launched by academia and industry towards this goal. Particularly, C4 chemicals such as succinic acid (SA) and 1,4-butanediol have been leading the path towards the commercialization of biobased technology with the effort of replacing chemical production. Here we present O-Succinyl-l-homoserine (SH) as a new, potentially important platform biochemical and demonstrate its central role as an intermediate in the production of SA, homoserine lactone (HSL), γ-butyrolactone (GBL) and its derivatives, and 1,4-butanediol (BDO). This technology encompasses (1) the genetic manipulation of Escherichia coli to produce SH with high productivity, (2) hydrolysis into SA and homoserine (HS) or homoserine lactone hydrochloride, and (3) chemical conversion of either HS or homoserine lactone HCL (HSL·HCl) into drop-in chemicals in polymer industry. This production strategy with environmental benefits is discussed in the perspective of targeting of fermented product and a process direction compared to petroleum-based chemical conversion, which may reduce the overall manufacturing cost.     

Sons and daughters beyond your control: episodes in the prehistory of the attention deficit/hyperactivity syndrome

Hyperactive and inattentive children have been discussed in both the pedagogic and medical literature since the nineteenth century, and many controversies associated with attention deficit and hyperactivity disorder (ADHD) have been repeatedly analyzed in different contexts. The ‘prehistory’ of the ADHD concept—that is, up to the definition of ADHD in DSM-III and of the corresponding ‘hyperkinetic disorder’ in ICD-9—is outlined, with an emphasis on the literature not previously discussed in English language reviews of the subject.     

Bacterial Diversity in the Rhizosphere of Cucumbers Grown in Soils Covering a Wide Range of Cucumber Cropping Histories and Environmental Conditions

Rhizosphere microorganisms in soils are important for plant growth. However, the importance of rhizosphere microorganisms is still underestimated since many microorganisms associated with plant roots cannot be cultured and since the microbial diversity in the rhizosphere can be influenced by several factors, such as the cropping history, biogeography, and agricultural practice. Here, we characterized the rhizosphere bacterial diversity of cucumber plants grown in soils covering a wide range of cucumber cropping histories and environmental conditions by using pyrosequencing of bacterial 16S rRNA genes. We also tested the effects of compost addition and/or bacterial inoculation on the bacterial diversity in the rhizosphere. We identified an average of approximately 8,883 reads per sample, corresponding to around 4,993 molecular operational taxonomic units per sample. The Proteobacteria was the most abundant phylum in almost all soils. The abundances of the phyla Bacteroidetes, Actinobacteria, Firmicutes, Acidobacteria, and Verrucomicrobia varied among the samples, and together with Proteobacteria, these phyla were the six most abundant phyla in almost all analyzed samples. Analyzing all the sample libraries together, the predominant genera found were Flavobacterium, Ohtaekwangia, Opitutus, Gp6, Steroidobacter, and Acidovorax. Overall, compost and microbial amendments increased shoot biomass when compared to untreated soils. However, compost addition decreased the bacterial α-diversity in most soils (but for three soils compost increased diversity), and no statistical effect of microbial amendment on the bacterial α-diversity was found. Moreover, soil amendments did not significantly influence the bacterial β-diversity. Soil organic content appeared more important than compost and microbial amendments in shaping the structure of bacterial communities in the rhizosphere of cucumber.     

Morphological forms of two macrophytes (yellow water-lily and arrowhead) along velocity gradient

We investigated morphological response of yellow water-lily and arrowhead to water velocity gradient in the lowland We?na river (Western Poland). Percentage cover of floating and submerged forms of yellow water-lily and arrowhead had been measured in randomly selected sites of 16 m². We analyzed 62 stands of Nuphar lutea and 80 stands of Sagittaria sagittifolia using 13 environmental variables. Redundancy analysis (RDA), the Monte Carlo permutation test, ranges of chemical and hydrological data, Generalized Additive Models (GAMs) and transfer function were used to describe reaction of investigated species to water velocity. Among the analyzed parameters of microhabitats with the floating and submerged forms of investigated species, velocity, pH, water colour, hydration and organic matter in bottom sediments were statistically significant. In S. sagittifolia case, the velocity was the most important parameter, while in N. lutea — both velocity and content of organic matter in river substrate were statistically significant.     

Geometric morphometric analysis of the freshwater prawn Macrobrachium borellii (Decapoda: Palaemonidae) at a microgeographical scale in a floodplain system

Morphometric studies are useful for delineating the shapes of various populations and species over geographical ranges and as evidence of regional differences in crustaceans. Hydrological fluctuations in a floodplain system modulate the dispersal and presence of decapods among habitats and constitute an important macrofactor that regulate other environmental variables and which could explain the richness, distribution and abundance of organisms that live in these systems. Morphological variations among populations of the freshwater prawn Macrobrachium borellii in a floodplain system at a microgeographical scale were studied using geometric morphometrics. Carapace structure was represented using 16 digitised landmarks. Allometry and sexual dimorphism was tested. Variation in shape was explored via Principal Component Analysis. Canonical Variate Analyses was applied to compare the differences in shape between species’ populations. The correlations and covariations among shapes and hydrometric level, current velocity, geographical location and hydrologic distances were analysed. The average carapace shape was different between sexes in all sites. Populations that were near each other in terms of hydrological distance had similar shapes, but all of the populations differed in shape from the farthest population. The environmental variables were not good predictors of the carapace shape. Instead, the shape was strongly related to the hydrologic distance and geographical location. The swimming characteristics of these prawns and their passive movements, together with the dynamics of a floodplain system, explain the low morphological variation between populations in this study. The dynamic characteristics of the system influence the dispersal of the prawns and allow populations to remain connected.     

Biosystematic studies of the Rumex acetosella complex V. Cytogeography and morphology in the Czech Socialistic Republic and a part of Lower Austria (Austria)

A cytogeographical and morphological (fruit-types and leaf-shape) study of the crítícal Rumex acetosella aggregate has been made on the basis of 38 population samples, for the most part from the Czech Socialistic Republic. In the genecological respect the populations from Bohemia and those from North and West Moravia fit into the pattern earlier found in West and Central Europe (i.e. they are 6x, mainly angiocarpous, and have non-dissected leaf lobes). Samples from the xerothermic region of South Moravia and parts of Slovakia and adjacent Austria are thought to be related to the Balkan population group (mainly 4x, gymnocarpous and with much dissected—multifid—leaf lobes). Evidence of introgressíve hybridisation from the 4x into the 6x ploidy-level has been found in the region of the well known migration route through the Moravian Gate.