Analyses of dryland biological soil crusts highlight lichens as an important regulator of microbial communities

Biological soil crusts (BSCs) provide important ecosystem services in dryland regions, including erosion control and contribution to nitrogen and CO₂ fixation. As soil microorganisms are still rarely studied within the context of biodiversity planning, we describe, as a contribution to the Soil Crust International project, an approach that addresses this gap in biodiversity assessments. The purpose of the present study was a characterization of prokaryotic communities of BSCs formed by two species of lichenized fungi, Psora decipiens and Toninia sedifolia, in relation to surrounding BSCs and the below-crust soil layer from Tabernas basin (Almería, Spain). Microbial community profiles were determined using 454 pyrosequencing targeting the V4 hypervariable region of the bacterial and archaeal 16S rRNA gene. The majority of the 65,497 sequences obtained belonged to Proteobacteria (mainly Alphaproteobacteria), Actinobacteria, Bacteroidetes and Cyanobacteria. Cyanobacteria were more abundant at the soil surface but rare in below-crust soils, whilst below-crust soils harbored significantly more Acidobacteria, Verrucomicrobia, Gemmatimonadetes, Planctomycetes and Armatimonadetes. Additionally, terricolous lichens were investigated using fluorescence in situ hybridization in conjunction with confocal laser scanning microscopy, the objective being to illustrate bacterial niches in BSC-forming lichens. Bacteria were mainly present at the upper cortex of the squamules and attachment organs. Our findings indicate that the composition of soil prokaryotes varies at a small scale not only in adjacent soil layers but also in BSC-forming lichen species. Furthermore, bacteria were shown to be attached to fungal structures, probably representing a case of fungal-bacterial interaction.     

Heterogeneity in ess transcriptional organization and variable contribution of the Ess/Type VII protein secretion system to virulence across closely related Staphylocccus aureus strains

The Type VII protein secretion system, found in Gram‐positive bacteria, secretes small proteins, containing a conserved W‐x‐G amino acid sequence motif, to the growth medium. Staphylococcus aureus has a conserved Type VII secretion system, termed Ess, which is dispensable for laboratory growth but required for virulence. In this study we show that there are unexpected differences in the organization of the ess gene cluster between closely related strains of S. aureus. We further show that in laboratory growth medium different strains of S. aureus secrete the EsxA and EsxC substrate proteins at different growth points, and that the Ess system in strain Newman is inactive under these conditions. Systematic deletion analysis in S. aureus RN6390 is consistent with the EsaA, EsaB, EssA, EssB, EssC and EsxA proteins comprising core components of the secretion machinery in this strain. Finally we demonstrate that the Ess secretion machinery of two S. aureus strains, RN6390 and COL, is important for nasal colonization and virulence in the murine lung pneumonia model. Surprisingly, however, the secretion system plays no role in the virulence of strain SA113 under the same conditions.     

Immune Evasion,Stress Resistance,and Efficient Nutrient Acquisition Are Crucial for Intracellular Survival of Candida glabrata within Macrophages

Candida glabrata is both a human fungal commensal and an opportunistic pathogen which can withstand activities of the immune system. For example, C. glabrata can survive phagocytosis and replicates within macrophages. However, the mechanisms underlying intracellular survival remain unclear. In this work, we used a functional genomic approach to identify C. glabrata determinants necessary for survival within human monocyte-derived macrophages by screening a set of 433 deletion mutants. We identified 23 genes which are required to resist killing by macrophages. Based on homologies to Saccharomyces cerevisiae orthologs, these genes are putatively involved in cell wall biosynthesis, calcium homeostasis, nutritional and stress response, protein glycosylation, or iron homeostasis. Mutants were further characterized using a series of in vitro assays to elucidate the genes'' functions in survival. We investigated different parameters of C. glabrata-phagocyte interactions: uptake by macrophages, replication within macrophages, phagosomal pH, and recognition of mutant cells by macrophages as indicated by production of reactive oxygen species and tumor necrosis factor alpha (TNF-α). We further studied the cell surface integrity of mutant cells, their ability to grow under nutrient-limited conditions, and their susceptibility to stress conditions mirroring the harsh environment inside a phagosome. Additionally, resistance to killing by neutrophils was analyzed. Our data support the view that immune evasion is a key aspect of C. glabrata virulence and that increased immune recognition causes increased antifungal activities by macrophages. Furthermore, stress resistance and efficient nutrient acquisition, in particular, iron uptake, are crucial for intraphagosomal survival of C. glabrata.     

Role of thylakoid Ndh complex and peroxidase in the protection against photo-oxidative stress: fluorescence and enzyme activities in wild-type and ndhF-deficient tobacco

An Ndh-deficient mutant of tobacco ( Nicotiana tabacum cv. Petit Havana) was prepared by disrupting the ndhF gene in a transplastomic approach. The mutant (Δ ndhF ) showed 10% of the Ndh complex activity (EC 1.6.5.3) and 8% of the NDH-F polypeptide of that of non-transformed plants. However, in Δ ndhF , NDH-A, another Ndh polypeptide, was still present at 50% of the level in non-transformed plants. Δ ndhF tobacco showed higher sensitivity than non-transformed plants to photo-oxidative stress (as judged by chlorophyll bleaching) caused by increased light intensity and paraquat applications. These photo-oxidative treatments increased the amount and activity of the Ndh complex, thylakoid peroxidase, post-illumination chlorophyll fluorescence and non-photochemical quenching (NPQ) of chlorophyll fluorescence in non-transformed but not in Δ ndhF tobacco. Highly stressed non-transformed plants showed a rapid post-rise decline of chlorophyll fluorescence, probably indicating a re-oxidation of reduced plastoquinone. The results indicate that, in normal plants, the Ndh complex and thylakoid peroxidase (EC 1.11.1.7) provide and remove electrons, respectively, to balance the redox level of the intermediates of cyclic electron transport. In this way, they optimize the generation of the transmembrane H⁺ gradient of thylakoids and, as a consequence, increase the NPQ and the protection against photo-oxidative stress.     

Intraspecific differences of Asian/Australian Phragmites australis subgroups reveal no potentially invasive traits

Hydrobiologia - Phragmites australis is a cosmopolitan plant species with high intraspecific diversity and phenotypic plasticity. Due to its variability and large ecological niche breadth,...     

The reproductive biology of Cyclops vicinus

Investigations of the reproductive biology of Cyclops vicinusrevealed that mated females oscillate between gravid and nongravidreproductive conditions. The gravid condition can be dividedin two recognizable phases: gravid/nonovigerous and gravid/ovigerous,the former phase being shortest. The maturation of new oocytestakes place when the old egg sacs are still being carried; thisensures a rapid clutch succession. Females which remain unmated,extrude few eggs, in no case complete egg sacs, and remain gravidthus conserving oocytes. Females which mate only once, showa similar reproductive pattern (clutch size and clutch succession)to those which remain combined with males, and thus have theopportunity to remate, but tend to produce fewer clutches. Malesare able to fertilize 3–4 females day^–1. Matingcapacity of males is possibly limited by the time needed tofill a new spermatophore. Short-term starvation (5 days) lengthenedclutch-to-clutch periods and diminished clutch size. When thestarvation period started in the gravid/ovigerous phase, a normalclutch was extruded but no new oocytes matured during starvation,indicating that the energy for egg maturations is provided inthe first part of the reproductive cycle.     

Use of space and behavior of weaned piglets kept in enriched two-level housing system

In this study, the possibility of introducing an elevated platform to a piglet pen was explored as a way of increasing available space and creating functional areas. On the platform, nine different manipulable materials were offered. In four batches, 40 weaned piglets were kept for five weeks in the two-level pen. Video recordings were taken two days per week. In the afternoon, more piglets were on the platform than in the morning or at night (7.2 ± 0.1 vs. 4.9 ± 0.1 vs. 0.6 ± 0.1 piglets/5 minutes; p < .05). The area under the platform was preferred more in the morning and at night than in the afternoon (18.5 ± 0.1 vs. 21.6 ± 0.2 vs. 12.5 ± 0.1 piglets/5 minutes; p < .05). Up to 36 piglets were counted there simultaneously, mainly in the recumbent position. On and under the platform, air velocity and ammonia concentration were within the recommended ranges. The study concluded that a two-level pen is a feasible option to increase space allowance and to create functional areas in a piglet pen.     

Experimental Sepsis Impairs Humoral Memory in Mice

Patients with sepsis are often immune suppressed, and experimental mouse models of sepsis also display this feature. However, acute sepsis in mice is also characterized by a generalized B cell activation and plasma cell differentiation, resulting in a marked increase in serum antibody concentration. Its effects on humoral memory are not clearly defined. We measured the effects of experimental sepsis on long-term immunological memory for a defined antigen: we induced colon ascendens stent peritonitis (CASP) 8 weeks after 2 rounds of immunization with ovalbumin. Four weeks later, the antigen-specific bone marrow plasma cell count had doubled in immunized non-septic animals, but remained unchanged in immunized septic animals. Sepsis also caused a decrease in antigen-specific serum antibody concentration. We conclude that sepsis weakens humoral memory by impeding the antigen-specific plasma cell pool’s development, which is not complete 8 weeks after secondary immunization.     

S100A11 plays a role in homologous recombination and genome maintenance by influencing the persistence of RAD51 in DNA repair foci

The repair of DNA double-strand breaks (DSBs) by homologous recombination (HR) is an essential process in maintenance of chromosomal stability. A key player of HR is the strand exchange factor RAD51 whose assembly at sites of DNA damage is tightly regulated. We detected an endogenous complex of RAD51 with the calcium-binding protein S100A11, which is localized at sites of DNA repair in HaCaT cells as well as in normal human epidermal keratinocytes (NHEK) synchronized in S phase. In biochemical assays, we revealed that S100A11 enhanced the RAD51 strand exchange activity. When cells expressing a S100A11 mutant lacking the ability to bind Ca²⁺, a prolonged persistence of RAD51 in repair sites and nuclear γH2AX foci was observed suggesting an incomplete DNA repair. The same phenotype became apparent when S100A11 was depleted by RNA interference. Furthermore, down-regulation of S100A11 resulted in both reduced sister chromatid exchange confirming the restriction of the recombination capacity of the cells, and in an increase of chromosomal aberrations reflecting the functional requirement of S100A11 for the maintenance of genomic stability. Our data indicate that S100A11 is involved in homologous recombination by regulating the appearance of RAD51 in DSB repair sites. This function requires the calcium-binding activity of S100A11.