Phylogeny of Microcystins: Evidence of a Biogeographical Trend?

Microcystins, the most prevalent cyanotoxins occurring worldwide, were first recorded in the species Microcystis aeruginosa. Its production has been reported in all continents; thus, we propose a comprehensive phylogenetic study to characterize M. aeruginosa microcystin-producing strains and establish whether or not the species has an historic biogeography. To accomplish this, we compared phylogenetically the nucleotide sequences of three genes of the mcy gene cluster (mcyA, mcyD and mcyG) from toxin producing M. aeruginosa strains across all the five continents. The obtained results provided valuable insight on the biogeography of M. aeruginosa produced microcystins: (i) the Asian strains showed to be distinct from the other continental groups indicating a genetically unique population and (ii) Asian strains were more related to European and North American strains. Moreover, the evidence of positive selection was determined in all the three mcy genes indicating that some functionality yet to be determined could be under selection for these genes.     

Activity of Ionotropic Glutamate Receptors in Retinal Cells: Effect of Ascorbate/Fe2+-Induced Oxidative Stress

Abstract: The effect of oxidative stress induced by the oxidant pair ascorbate/Fe²⁺ on the activity of ionotropic glutamate receptors was studied in cultured chick retina cells. The release of [³H]GABA and the increase of the intracellular free Na⁺ concentration ([Na⁺]_i), evoked by glutamate receptor agonists, were used as functional assays for the activity of the receptors. The results show that the maximal release of [³H]GABA evoked by kainate (KA; ～20% of the total) or AMPA (～11% of the total) was not different in control and peroxidized cells, whereas the EC₅₀ values determined for peroxidized cells (33.6 ± 1.7 and 8.0 ± 2.0 µM for KA and AMPA, respectively) were significantly lower than those determined under control conditions (54.1 ± 6.6 and 13.0 ± 2.2 µM for KA and AMPA, respectively). The maximal release of [³H]GABA evoked by NMDA under K⁺ depolarization was significantly higher in peroxidized cells (7.5 ± 0.5% of the total) as compared with control cells (4.0 ± 0.2% of the total), and the effect of oxidative stress was significantly reduced by a phospholipase A₂ inhibitor or by fatty acid-free bovine serum albumin. The change in the intracellular [Na⁺]_i evoked by saturating concentrations of NMDA under depolarizing conditions was significantly higher in peroxidized cells (8.9 ± 0.6 mM) than in control cells (5.9 ± 1.0 mM). KA, used at a subsaturating concentration (35 µM), evoked significantly greater increases of the [Na⁺]_i in peroxidized cells (11.8 ± 1.7 mM) than in control cells (7.1 ± 0.8 mM). A saturating concentration (150 µM) of this agonist triggered similar increases of the [Na⁺]_i in control and peroxidized cells. Accordingly, the maximal number of binding sites for (+)-5-[³H]methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine maleate ([³H]MK-801) was increased after peroxidation, whereas the maximal number of binding sites for [³H]KA was not affected by oxidative stress. These data suggest that under oxidative stress the activity of the ionotropic glutamate receptors is increased, with the NMDA receptor being the most affected by peroxidation.     

The importance of qualitative inventory sampling using electric fishing and nets in a large,tropical river (Brazil)

Qualitative fish sampling (single catch) using three methods (electric fishing, gillnets, seine nets) was assessed at three sites in the Ivai River (Paraná State, Brazil) to check their usefulness for quick inventory investigations. Electric fishing at a constant effort (15 min per each) was considered to be the best sampling technique. Taxon richness was calculated as the expected number of species using a rarefraction technique. Samples of 300–325 individuals, and 6–10 repetitions in neighbouring segments were sufficient. Even though electric fishing was conducted with the same effort in similar segments at each site there were many differences in species abundance. This suggests that the estimated fish specimen number obtained with constant effort is of limited validity on one sampling occasion and may be best considered as an ‘index of density’ only. The differences may partially be caused by the segments being located along the right and left bank of the large river (problem of different habitats). Qualitative and quantitative differences between electric fishing, gill-netting and seine-netting samples were very high at a high significance level. Nevertheless, as 22.4% taxa were caught only by gill or seine nets, these gears were important for complementing the species list. This revised version was published online in July 2006 with corrections to the Cover Date.     

Detection of cytokinins and auxin in plant tissues using histochemistry and immunocytochemistry

We report a new method for histochemical localization of cytokinins (CKs) in plant tissues based on bromophenol blue/silver nitrate staining. The method was validated by immunohistochemistry using anti-trans-zeatin riboside antibody. Indole-3-acetic acid (auxin, IAA) was localized by anti-IAA antibody in plant tissues as a proof for IAA histolocalization. We used root sections, because they are major sites of CKs synthesis, and insect galls of Piptadenia gonoacantha that accumulate IAA. Immunostaining confirmed the presence of zeatin and sites of accumulation of IAA indicated by histochemistry. The colors developed by histochemical reactions in free-hand sections of plant tissues were similar to those obtained by thin layer chromatography (TLC), which reinforced the reactive sites of zeatin. The histochemical method for detecting CKs is useful for galls and roots, whereas IAA detection is more efficient for gall tissues. Therefore, galls constitute a useful model for validating histochemical techniques due to their rapid cell cycles and relatively high accumulation of plant hormones.     

A Lectin-like Molecule is Discharged from Mucocysts of Tetrahymena pyriformis in the Presence of Insulin

ABSTRACT. By use of a monoclonal antibody directed against purified lectin from the sponge Geodia cydonium it was demonstrated that the mucocysts of Tetrahymena pyriformis contain a substance immunologically similar to that found in G. cydonium . In extracts of T. pyriformis the monoclonal antibody recognizes a 36 kDa protein; binding could be abolished by adsorption of the antibody with (i) crude extract, (ii) purified lectin from G. cydonium and (iii) a 29 aa long peptide. In addition the data show that 10^-6 M of insulin causes first the release of mucocyst material, which reacts with the lectin antibody, and second its subsequent redistribution on the surface of the somatic cilia and the oral field.     

The dependence of fish community structure and dynamics on floodplain and riparian ecotone zone in Parana River,Brazil

Flood intensity was a main factor determining access to the riparian/floodplain ecotonal resources of the upper Parana River, Brazil, and in consequence determining yield of the dominant trophic groups, which are fish feeding on flooded grasslands and on riparian fruits and leaves. Prey and predator density both declined in years of low floods, and predators did not recover until the next year of high flood, more slowly than in the case prey fish, most probably due to differences in life cycle length. The life cycle of one of the most important species, Curimba (Prochilodus lineatus — Characidae) depends on access to the floodplain lagoons and return to the reservoir after maturation for intensive growth. The riparian/floodplain habitat complexity and connectivity has great significance for fish community dynamics and fisheries yield, and may serve a reason to create a national park in the last floodplain section of the upper Parana River.     

Comparison of pome fruit orchard inhabiting spider assemblages at different geographical scales

1 The composition of pome fruit orchard inhabiting spider assemblages was investigated at different geographical scales (Holarctic, European, inter- and intraregional levels within Hungary) using previous faunistic studies and data collected in Hungary between 1995 and 1997. Samples in Hungary were taken from the canopy and herb layer of apple and pear orchards in five markedly different fruit-growing regions by beating and sweep-netting methods. 2 The composition of canopy spider assemblages of apple orchards was analysed for the Holartic region and found to be determined by latitude at family level, and by the main zoogoegraphical regions at genus level. At the European scale, both the genus and species composition changed along a north–south gradient. 3 A comparison among apple and pear orchards located in different regions in Hungary, showed that both foliage- and grass-dwelling spider assemblages varied considerably in species composition and dominance order. 4 Within the same region, both the foliage- and grass-dwelling spider assemblages showed moderate differences in apple and pear orchards submitted to different treatments. Although the assemblages of spiders inhabiting the canopy and the herbaceous layer can be unambiguously distinguished, some overlap still occurs. 5 We conclude that the composition of spider assemblages is basically determined by geographical location. Although both pesticide treatments and available prey densities can influence the population of spiders, such factors are of moderate importance when compared with the effect of regionality, even when considered at smaller scale. However, most members of the family Theridiidae and the large orb-weavers (Araneidae) decreased considerably in treated plots. Scale-specific differences are thus relevant in determining the composition of prey–predator systems in orchards, and should be taken into account when designing integrated pest management (IPM) programs for apple and pear orchards.     

Genetic characterization of <Emphasis Type="Italic">Microcystis aeruginosa</Emphasis> isolates from Portuguese freshwater systems

Cyanobacteria are microorganisms that pose a serious threat to the aquatic waterways through the production of dense blooms under eutrophic conditions and the release of toxic secondary metabolites—cyanotoxins. Within cyanobacteria, the colonial planktonic Microcystis aeruginosa is widely distributed in both fresh and brackish aquatic environments throughout the world being frequently observed in the Portuguese water systems. Apart from the well-established distribution of M. aeruginosa in Portugal, knowledge of its genetic diversity and population structure is unknown. Therefore, in this study twenty-seven strains were obtained from the North, Centre and South regions of Portugal and were subjected to extensive phylogenetic analyses using simultaneously four distinct genetic markers (16S rRNA, 16S-23S ITS, DNA gyrase subunit ß and cell division protein (ftsZ)) encompassing in total 2834 bp. With this work we characterized the phylogenetic relationship among the Portuguese strains, with the southern strains showing higher genetic structure relatively to the North and Centre strains. A total of fifteen genotypes were determined for M. aeruginosa in Portuguese water systems revealing a high genetic diversity. This is also the first study to report geographic variation on the population structure of the Portuguese M. aeruginosa.     

Phylogeography of the heavily poached African common pangolin (Pholidota,Manis tricuspis) reveals six cryptic lineages as traceable signatures of Pleistocene diversification

Knowledge on faunal diversification in African rainforests remains scarce. We used phylogeography to assess (i) the role of Pleistocene climatic oscillations in the diversification of the African common pangolin (Manis tricuspis) and (ii) the utility of our multilocus approach for taxonomic delineation and trade tracing of this heavily poached species. We sequenced 101 individuals for two mitochondrial DNA (mtDNA), two nuclear DNA and one Y‐borne gene fragments (totalizing 2602 bp). We used a time‐calibrated, Bayesian inference phylogenetic framework and conducted character‐based, genetic and phylogenetic delineation of species hypotheses within African common pangolins. We identified six geographic lineages partitioned into western Africa, Ghana, the Dahomey Gap, western central Africa, Gabon and central Africa, all diverging during the Middle to Late Pleistocene. MtDNA (cytochrome b + control region) was the sole locus to provide diagnostic characters for each of the six lineages. Tree‐based Bayesian delimitation methods using single‐ and multilocus approaches gave high support for ‘species’ level recognition of the six African common pangolin lineages. Although the diversification of African common pangolins occurred during Pleistocene cyclical glaciations, causative correlation with traditional rainforest refugia and riverine barriers in Africa was not straightforward. We conclude on the existence of six cryptic lineages within African common pangolins, which might be of major relevance for future conservation strategies. The high discriminative power of the mtDNA markers used in this study should allow an efficient molecular tracing of the regional origin of African common pangolin seizures.