The possible influence of LuxS in the in vivo virulence of rabbit enteropathogenic Escherichia coli

Attaching and effacing (A/E) organisms, such as rabbit enteropathogenic Escherichia coli (EPEC), human EPEC or enterohemorrhagic E. coli (EHEC) share attaching and effacing phenotype and LEE pathogenicity island responsible for A/E. The present study was undertaken to investigate the impact of the LuxS quorum sensing (QS) signaling system in vitro and in vivo pathogenicity of A/E organisms using rabbit EPEC (rEPEC) strain E22 (O103:H2). Analysis of the bioluminescence indicated abolished production of the QS signal AI-2 by luxS mutant (E22DeltaluxS). Strain E22Deltalux also exhibited impaired expression of several normally secreted proteins and reduced adherence to cultured HeLa cells. Complementation of the intact luxS gene to E22DeltaluxS restored secreted protein expression comparable to the WT type but not adherence to HeLa cells. In experimentally infected rabbits, the isogenic luxS mutant induced clinical illness and intimate adherence to the intestinal mucosa, albeit to a less extent, comparable to that seen with the parent virulent strain. It is worth noting that reduced fecal bacterial shedding, mucosal adherence and improved cumulative weight gain were seen for the mutant strain complemented with luxS when compared to the WT. It appears that the luxS gene is not essential for in vivo pathogenicity by rEPEC where exogenous QS signals are present in the gut. The impact of AI-2 provided by multicopy plasmid on bacterial virulence is discussed.     

Modelling diapause termination and phenology of the Japanese beetle,Popillia japonica

We deve?loped a mechanistic, stage-structured model simulating the phenology of Popillia japonica. The model simulates the influence of soil temperature on the larval diapause termination and on the development rate function of post-overwintering larvae and pupae. Model parameters are estimated based on literature evidence for pupae development and on a parameterisation process that allows estimating parameters for larval diapause termination and for the development rate function (and the related uncertainty) of post-overwintering larvae. Data used for model parameterisation and validation refer to time-series adult trap catches collected during the P. japonica monitoring programme performed by the Phytosanitary Service of Lombardy Region within the infested area in Lombardy (Italy) from 2015 to 2019. A total of 12 randomly selected locations are used to estimate biologically realistic model parameters (parameterisation dataset). We applied a Jackknife nonparametric resampling procedure on the parameterisation dataset to quantify uncertainty associated with parameters’ estimates. Parameterised model is then validated on time-series adult trap catches data referring to a different set of 12 randomly selected locations (validation dataset) surveyed in Lombardy. The model successfully predicted the beginning of adult emergence and the overall curve of adult emergence in the validation dataset. The model presented can support the definition of the best timing for the implementation of monitoring and control activities for the local and the area-wide management of P. japonica.

     

NITRATE UPTAKE KINETICS AND METABOLIC PARAMETERS IN TWO RICE VARIETIES GROWN IN HIGH AND LOW NITRATE

A study was conducted on the effect of nitrate (NO₃) levels on nitrate uptake kinetics and nitrogen (N) metabolism in two rice varieties, Piaui (landrace) and IAC-47 (improved). At 27 days after germination (DAG) N supply was suspended for 72 h, and then restored as 0.2 or 2 mM nitrate. The nitrate uptake kinetics was determined by the depletion method. Plants were harvested at 0, 6, and 24 h. Plants of the Piaui variety under 0.2 mM nitrate showed higher V_max and lower K_M, indicating higher efficiency of nitrate uptake at low supply. In the sheaths of both varieties, there was a greater accumulation of nitrate and lower activities of nitrate reductase and glutamine synthetase. The V-H⁺-ATPase activity increased between 0 and 6 h accompanying the nitrate influx, suggesting that the activity of this proton pump is necessary for the antiport activity (H⁺/2NO^? ₃) involved in the accumulation of nitrate into vacuoles.     

Targeting QseC signaling and virulence for antibiotic development

Many bacterial pathogens rely on a conserved membrane histidine sensor kinase, QseC, to respond to host adrenergic signaling molecules and bacterial signals in order to promote the expression of virulence factors. Using a high-throughput screen, we identified a small molecule, LED209, that inhibits the binding of signals to QseC, preventing its autophosphorylation and consequently inhibiting QseC-mediated activation of virulence gene expression. LED209 is not toxic and does not inhibit pathogen growth; however, this compound markedly inhibits the virulence of several pathogens in vitro and in vivo in animals. Inhibition of signaling offers a strategy for the development of broad-spectrum antimicrobial drugs.     

Production of selected short-rotation wood crop species and quality of obtained biomass

Short-rotation coppice (SRC) represents an important source of wood biomass. Many uncertainties create barriers to farmers establishing SRC plantations, especially under Mediterranean climate conditions. In this study, five species and respective genotypes were analyzed. The five species were Fraxinus angustifolia, Robinia pseudoacacia, Salix alba, Populus nigra (Limatola), and Populus × euroamericana, with the genotypes of the latter being Grimminge, Vesten, Hoogvorst, Muur. For the plantations studied, two different harvesting systems were replicated: the single machine pass, or cut-and-chip (CC); and the double machine pass, for which the tree are cut first, then chipped later (chip of stored trees CS). In the CC, fresh trees were harvested and chipped by Claas Jaguar 880. In the CS, dry trees were chipped by Farmi Forest CH 260. Within the same site, in climatic conditions and low-input management, the best result in terms of biomass yield was obtained from the black poplar Limatola. Both wood typologies and harvesting systems affected the chip quality. The disk chipper, when working on dried biomass, produced lower quality chips than the other chipping device in terms of particle size. In the chips obtained, there was an increase in the number of chips classified as being “larger size” and “oversized”, and a decrease in the percentage of those classified as “accept” (45–3 mm fraction). The chips obtained from dried trees were of better energy quality compared with the same biomass obtained from the fresh trees in terms of heating value and ash content.