Role of glucose and CcpA in capsule expression and virulence of Streptococcus suis

Streptococcus suis is one of the most important pathogens in pigs and is also an emerging zoonotic agent. After crossing the epithelial barrier, S. suis causes bacteraemia, resulting in meningitis, endocarditis and bronchopneumonia. Since the host environment seems to be an important regulatory component for virulence, we related expression of virulence determinants of S. suis to glucose availability during growth and to the sugar metabolism regulator catabolite control protein A (CcpA). We found that expression of the virulence-associated genes arcB, representing arcABC operon expression, cps2A, representing capsular locus expression, as well as sly, ofs, sao and epf, differed significantly between exponential and early stationary growth of a highly virulent serotype 2 strain. Deletion of ccpA altered the expression of the surface-associated virulence factors arcB, sao and eno, as well as the two currently proven virulence factors in pigs, ofs and cps2A, in early exponential growth. Global expression analysis using a cDNA expression array revealed 259 differentially expressed genes in early exponential growth, of which 141 were more highly expressed in the CcpA mutant strain 10¿ccpA and 118 were expressed to a lower extent. Interestingly, among the latter genes, 18 could be related to capsule and cell wall synthesis. Correspondingly, electron microscopy characterization of strain 10¿ccpA revealed a markedly reduced thickness of the capsule. This phenotype correlated with enhanced binding to porcine plasma proteins and a reduced resistance to killing by porcine neutrophils. Taken together, our data demonstrate that CcpA has a significant effect on the capsule synthesis and virulence properties of S. suis

Sequence of a putative human housekeeping gene (HK33) localized on chromosome 1

A gene (X33) localized on human chromosome 1 has been detected by crossreaction of its fusion protein with a monospecific antiserum directed against human vitamin-D-binding protein (hDBP; group-specific component). Its cDNA sequence analysis showed no evident homologies neither to the sequence encoding hDBP nor to any other sequence. The largest cDNA clone of 3.2 kb includes a 897-bp coding region and a large 3’ untranslated region with at least four polyadenylation sites. Further cDNA amplification using PCR demonstrated a total cDNA length of approx. 3.7 kb. Northern blot analysis revealed signals at about 2.2-2.5 kb and 4.0 kb, the shorter transcripts representing mRNAs using one of the two polyadenylation sites at about 2.0 kb. Synthesis of the 299-amino-acid polypeptide (33 kDa) in the bacterial host, with subsequent Western blot analysis, verified the sequence-specific recognition by the hDBPspecific antiserum. The search of protein databanks revealed no homology of HK33 to any known sequence. Since the gene is transcribed in all cells and tissues tested so far, it is a strong candidate for another housekeeping gene

Nitrous oxide production in soil isolates of nitrate-ammonifying bacteria

Here we provide the first demonstration of the potential for N2O production by soil-isolated nitrate-ammonifying bacteria under different C and N availabilities, building on characterizations informed from model strains. The potential for soil-isolated Bacillus sp. and Citrobacter sp. to reduce NO3-, and produce NH4+, NO2- and N2O was examined in batch and continuous (chemostat) cultures under different C-to-NO3- ratios, NO3--limiting (5 mM) and NO3--sufficient (22 mM) conditions. C-to-NO3- ratio had a major influence on the products of nitrate ammonification, with NO2-, rather than NH4+, being the major product at low C-to-NO3- ratios in batch cultures. N2O production was maximum and accompanied by high NO2- production under C-limitation/NO3-sufficiency conditions in chemostat cultures. In media with lower C-to-NO3-N ratios (5- and 10-to-1) up to 2.7% or 5.0% of NO3- was reduced to N2O by Bacillus sp. and Citrobacter sp., respectively, but these reduction efficiencies were only 0.1% or 0.7% at higher C-to-NO3- ratios (25- and 50-to-1). As the highest N2O production did not occur under the same C-to-NO3- conditions as highest NH4+ production we suggest that a re-evaluation may be necessary of the environmental conditions under which nitrate ammonification contributes to N2O emission from soil

Hydrodynamic controls on alluvial ridge construction and avulsion likelihood in meandering river floodplains

This is the author accepted manuscript. The final version is available from Geological Society of America via the DOI in this record.Existing models of alluvial stratigraphy often neglect the hydrodynamic controls on channel belt and floodplain sedimentation, and predict avulsion using topographic metrics, such as channel belt super-elevation (the ratio of alluvial ridge height to channel depth). This study provides a first demonstration of the potential for simulating long-term river floodplain evolution (over >500 floods) using a process-based hydrodynamic model. Simulations considered alluvial ridge construction during the period leading up to an avulsion, and assess the controls on avulsion likelihood. Results illustrate that the balance between within-channel and overbank sedimentation exerts a key control on both super-elevation ratios and on the conveyance of water and sediment to the floodplain. Rapid overbank sedimentation creates high alluvial ridges with deep channels, leading to lower apparent super-elevation ratios, and implying a reduced likelihood of avulsion. However, channel deepening also drives a reduction in channel belt–floodplain connectivity, so that conveyance of water to the distal floodplain is concentrated in a declining number of channel breaches, which may favor avulsion. These results suggest that, while super-elevation ratios in excess of a threshold value may be a necessary condition for a meandering river avulsion, the likelihood of avulsion may not be greatest where the super-elevation ratio is maximized. Instead, optimal conditions for avulsion may depend on channel-floodplain hydrodynamic connectivity, determined by the balance between coarse (channel bed–forming) and fine (floodplain-constructing) sediment delivery. These results highlight a need to rethink the representation of avulsion in existing models of alluvial architecture.This work was funded by UK Natural Environment Research Council grants NE/H009108/1 and NE/H007288/1

454-Pyrosequencing: A Molecular Battiscope for Freshwater Viral Ecology

Viruses, the most abundant biological entities on the planet, are capable of infecting organisms from all three branches of life, although the majority infect bacteria where the greatest degree of cellular diversity lies. However, the characterization and assessment of viral diversity in natural environments is only beginning to become a possibility. Through the development of a novel technique for the harvest of viral DNA and the application of 454 pyrosequencing, a snapshot of the diversity of the DNA viruses harvested from a standing pond on a cattle farm has been obtained. A high abundance of viral genotypes (785) were present within the virome. The absolute numbers of lambdoid and Shiga toxin (Stx) encoding phages detected suggested that the depth of sequencing had enabled recovery of only ca. 8% of the total virus population, numbers that agreed within less than an order of magnitude with predictions made by rarefaction analysis. The most abundant viral genotypes in the pond were bacteriophages (93.7%). The predominant viral genotypes infecting higher life forms found in association with the farm were pathogens that cause disease in cattle and humans, e.g. members of the Herpesviridae. The techniques and analysis described here provide a fresh approach to the monitoring of viral populations in the aquatic environment, with the potential to become integral to the development of risk analysis tools for monitoring the dissemination of viral agents of animal, plant and human diseases

Modelling flow through a permeable bed: a combined physical-numerical approach

River hydrodynamicsTurbulent open channel flow and transport phenomen