Variability in molecular characteristics of Hepatitis E virus quasispecies could modify viral surface properties and transmission.

Hepatitis E virus (HEV) usually causes self-limited liver diseases but can also result in severe cases. Genotypes 1 (G1) and 2 circulate in developing countries are human-restricted and waterborne, while zoonotic G3 and G4 circulating in industrialized countries preferentially infect human through consumption of contaminated meat. Our aims were to identify amino acid patterns in HEV variants that could be involved in pathogenicity or in transmission modes, related to their impact on antigenicity and viral surface hydrophobicity. HEV sequences from human (n = 37) and environmental origins (wild boar [n = 3], pig slaughterhouse effluent [n = 6] and urban wastewater [n = 2]) were collected for the characterization of quasispecies using ultra-deep sequencing (ORF2/ORF3 overlap). Predictive and functional assays were carried out to investigate viral particle antigenicity and hydrophobicity. Most quasispecies showed a major variant while a mixture was observed in urban wastewater and in one chronically infected patient. Amino acid signatures were identified, as a rabbit-linked HEV pattern in two infected patients, or the S68L (ORF2) / H81C (ORF3) residue mostly identified in wild boars. By comparison with environmental strains, molecular patterns less likely represented in humans were identified. Patterns impacting viral hydrophobicity and/or antigenicity were also observed, and the higher hydrophobicity of HEV naked particles compared with the enveloped forms was demonstrated. HEV variants isolated from human and environment present molecular patterns that could impact their surface properties as well as their transmission. These molecular patterns may concern only one minor variant of a quasispecies and could emerge under selective pressure.

Inactivation of murine norovirus and hepatitis A virus on fresh raspberries by gaseous ozone treatment.

Raspberries are vulnerable products for which industrial treatment solutions ensuring both food safety and sensory quality are not easily applicable. Raspberries have been associated with numerous foodborne outbreaks in recent decades. Ozone has been proven effective as a drinking water treatment against pathogenic microorganisms. Nevertheless, to date, little information is available regarding the effect of gaseous ozone on viruses in food matrices. A comparison of the effect of gaseous ozone on murine norovirus (MNV-1) and hepatitis A virus (HAV) adsorbed on fresh raspberries was performed. Infectious MNV-1 was highly inactivated (>3.3 log10) by ozone (3 ppm, 1 min). The raspberry matrix seems to enhance inactivation by ozone compared to water. The same treatment was observed to have little effect on HAV even for the highest dose under the tested conditions (5 ppm, 3 min). Ozone treatment (5 ppm, 3 min) did not affect the appearance of raspberries even after three days post-treatment. No ozone effect was observed on the genomes detected by RT-PCR on both tested viruses, irrespective of the matrix or tested doses used. Gaseous ozone could therefore be a good candidate for human norovirus inactivation on raspberries but new conditions are needed for it to have significant effects on HAV inactivation.

Functional breadth and home-field advantage generate functional differences among soil microbial decomposers.

In addition to the effect of litter quality (LQ) on decomposition, increasing evidence is demonstrating that carbon mineralization can be influenced by the past resource history, mainly through following two processes: (1) decomposer communities from recalcitrant litter environments may have a wider functional ability to decompose a wide range of litter species than those originating from richer environments, i.e., the functional breadth (FB) hypothesis; and/or (2) decomposer communities may be specialized towards the litter they most frequently encounter, i.e., the home-field advantage (HFA) hypothesis. Nevertheless, the functional dissimilarities among contrasting microbial communities, which are generated by the FB and the HFA, have rarely been simultaneously quantified in the same experiment, and their relative contributions over time have never been assessed. To test these hypotheses, we conducted a reciprocal transplant decomposition experiment under controlled conditions using litter and soil originating from four ecosystems along a land-use gradient (forest, plantation, grassland, and cropland) and one additional treatment using 13C-labelled flax litter allowing us to assess the priming effect (PE) in each ecosystem. We found substantial effects of LQ on carbon mineralization (more than two-thirds of the explained variance), whereas the contribution of the soil type was fairly low (less than one-tenth), suggesting that the contrasting soil microbial communities play only a minor role in regulating decomposition rates. Although the results on PE showed that we overestimated litter-derived CO2 fluxes, litter-microbe interactions contributed significantly to the unexplained variance observed in carbon mineralization models. The magnitudes of FB and HFA were relatively similar, but the directions of these mechanisms were sometimes opposite depending on the litter and soil types. FB and HFA estimates calculated on parietal sugar mass loss were positively correlated with those calculated on enzymatic activity, confirming the idea that the interaction between litter quality and microbial community structure may modify the trajectory of carbon mineralization via enzymatic synthesis. We conclude that although litter quality was the predominant factor controlling litter mineralization, the local microbial communities and interactions with their substrates can explain a small (< 5%) but noticeable portion of carbon fluxes.

Improving selectivity of dopamine D3 receptor ligands.

The seminal human dopamine D3 receptor (hD3R) ligand BP 897 has shown interesting properties during clinical trials. However, its lack of selectivity towards human adrenergic receptor impedes further development. Two approaches were followed to increase hD3R selectivity. The lead optimisation succeeded, we disclose here ligands with subnanomolar potency for D3R, combined with a good selectivity for the closely related human dopamine D2 and human adrenergic alpha-1 receptors.

Synthesis and evaluation of a 2-benzothiazolylphenylmethyl ether class of histamine H4 receptor antagonists.

Synthesis and biological evaluation of a new class of histamine H4 receptor ligands, distinct from the previously reported chemotypes, are described. A virtual screening of our corporate compound collection identified a hit with an undesired dual H3R/H4R activity. Chemical exploration led to the discovery of a more potent and selective 2-benzothiazolylphenylmethyl ether lead compound.

The societal impact of herpes zoster and postherpetic neuralgia on patients, life partners, and children of patients in Germany.

The aim of this study was to assess the impact of herpes zoster (HZ) and postherpetic neuralgia (PHN) on the daily activities of patients and family members who care for them. Some former patients and family members participated in face-to-face interviews or in a T-group meeting (qualitative phase) and some participated in telephone interviews (quantitative phase). They all expressed feelings of helplessness and frustration mixed with depression, sadness, or rage. Many of the former patients said their lives stopped, in contrast to family members who said that their lives were busy and stressful. Family members caring for patients with PHN were more psychologically stressed than those caring for patients with HZ. Although former patients appreciated the psychological and emotional support given by their family members, they underestimated the impact that their disease had on them. Former patients and their family never forgot this illness and its considerable impact on their lives, particularly when PHN occurred. We need to raise the awareness of the general public about the real life impact of HZ and PHN and their often severe, debilitating consequences and the potential benefits from vaccination.

Earthworms and plants can decrease soil greenhouse gas emissions by modulating soil moisture fluctuations and soil macroporosity in a mesocosm experiment.

Earthworms can stimulate microbial activity and hence greenhouse gas (GHG) emissions from soils. However, the extent of this effect in the presence of plants and soil moisture fluctuations, which are influenced by earthworm burrowing activity, remains uncertain. Here, we report the effects of earthworms (without, anecic, endogeic, both) and plants (with, without) on GHG (CO2, N2O) emissions in a 3-month greenhouse mesocosm experiment simulating a simplified agricultural context. The mesocosms allowed for water drainage at the bottom to account for the earthworm engineering effect on water flow during two drying-wetting cycles. N2O cumulative emissions were 34.6% and 44.8% lower when both earthworm species and only endogeic species were present, respectively, and 19.8% lower in the presence of plants. The presence of the endogeic species alone or in combination with the anecic species slightly reduced CO2 emissions by 5.9% and 11.4%, respectively, and the presence of plants increased emissions by 6%. Earthworms, plants and soil water content interactively affected weekly N2O emissions, an effect controlled by increased soil dryness due to drainage via earthworm burrows and mesocosm evapotranspiration. Soil macroporosity (measured by X-ray tomography) was affected by earthworm species-specific burrowing activity. Both GHG emissions decreased with topsoil macropore volume, presumably due to reduced moisture and microbial activity. N2O emissions decreased with macropore volume in the deepest layer, likely due to the presence of fewer anaerobic microsites. Our results indicate that, under experimental conditions allowing for plant and earthworm engineering effects on soil moisture, earthworms do not increase GHG emissions, and endogeic earthworms may even reduce N2O emissions.

Assessing infectivity of emerging enveloped viruses in wastewater and sewage sludge: Relevance and procedures.

The emergence of SARS-CoV-2 has heightened the need to evaluate the detection of enveloped viruses in the environment, particularly in wastewater, within the context of wastewater-based epidemiology. The studies published over the past 80 years focused primarily on non-enveloped viruses due to their ability to survive longer in environmental matrices such as wastewater or sludge compared to enveloped viruses. However, different enveloped viruses survive in the environment for different lengths of time. Therefore, it is crucial to be prepared to assess the potential infectious risk that may arise from future emerging enveloped viruses. This will require appropriate tools, notably suitable viral concentration methods that do not compromise virus infectivity. This review has a dual purpose: first, to gather all the available literature on the survival of infectious enveloped viruses, specifically at different pH and temperature conditions, and in contact with detergents; second, to select suitable concentration methods for evaluating the infectivity of these viruses in wastewater and sludge. The methodology used in this data collection review followed the systematic approach outlined in the PRISMA (Preferred Reporting Items for Systematic Review and Meta-Analysis) guidelines. Concentration methods cited in the data gathered are more tailored towards detecting the enveloped viruses' genome. There is a lack of suitable methods for detecting infectious enveloped viruses in wastewater and sludge. Ultrafiltration, ultracentrifugation, and polyethylene glycol precipitation methods, under specific/defined conditions, appear to be relevant approaches. Further studies are necessary to validate reliable concentration methods for detecting infectious enveloped viruses. The choice of culture system is also crucial for detection sensitivity. The data also show that the survival of infectious enveloped viruses, though lower than that of non-enveloped ones, may enable environmental transmission. Experimental data on a wide range of enveloped viruses is required due to the variability in virus persistence in the environment.

A new understanding of somatic coliphages belonging to the Microviridae family in urban wastewater.

Somatic coliphages (SC) and F-specific RNA coliphages (FRNAPH) have been included in regulations or guidelines by several developed countries as a way of monitoring water safety and the microbiological quality of shellfish harvesting waters. SC are highly diverse in their morphology, size and genome. The Microviridae family contains three genera of phages (Alphatrevirus, Gequatrovirus, and Sinsheimervirus), all having a capsid of similar morphology (icosahedral) and size (25-30 nm in diameter) to that of common pathogenic enteric viruses. Three PCR assays specific for each genus of Microviridae were designed to study these phages in raw and treated wastewater (WW) in order to gain knowledge about the diversity and prevalence of Microviridae among SC, as well as their inactivation and removal during WW treatments. Among the four wastewater treatment plants (WWTPs) monitored here, two WWTPs applied disinfection by UV light as tertiary treatment. First, we noticed that Microviridae represented 10 to 30 % of infectious SC in both raw and treated WW. Microviridae appeared to behave in the same way as all SC during these WW treatments. As expected, the highest inactivation, at least 4 log10, was achieved for infectious Microviridae and SC in both WWTPs using UV disinfection. PCR assays showed that the highest removal of Microviridae reached about 4 log10, but the phage removal can vary greatly between WWTPs using similar treatments. This work forms the basis for a broader evaluation of Microviridae as a viral indicator of water treatment efficiency and WW reuse.

Novel and highly potent histamine H3 receptor ligands. Part 3: an alcohol function to improve the pharmacokinetic profile.

Synthesis and biological evaluation of potent histamine H3 receptor antagonists incorporating a hydroxyl function are described. Compounds in this series exhibited nanomolar binding affinities for human receptor, illustrating a new possible component for the H3 pharmacophore. As demonstrated with compound BP1.4160 (cyclohexanol 19), the introduction of an alcohol function counter-intuitively allowed to reach high in vivo efficiency and favorable pharmacokinetic profile with reduced half-life.

Determination of the binding mode and interacting amino-acids for dibasic H3 receptor antagonists.

Due to its involvement in major CNS functions, the histamine H3 receptor (H3R) is the subject of intensive medicinal chemistry investigation, supported by the range of modern drug discovery tools, such as receptor modeling and ligand docking. Although the receptor models described to date share a majority of common traits, they display discrete alternatives in amino-acid conformation, rendering ligand binding modes quite different. Such variations impede structure-based drug design in the H3R field. In the present study, we used a combination of medicinal chemistry, receptor-guided and ligand-based methods to elucidate the binding mode of antagonists. The approaches converged towards a ligand orientation perpendicular to the membrane plane, bridging Glu206 of the transmembrane helix 5 to acidic amino acids of the extracellular loops. This consensus will help future structure-based drug design for H3R ligands.

Two-day detection of infectious enteric and non-enteric adenoviruses by improved ICC-qPCR.

In order to provide a more suitable response to public health concerns, we improved the detection of infectious human adenoviruses in water by optimising the commonly used integrated cell culture-PCR method. Risk evaluation studies seek for rapid detection of infectious adenoviruses, including the enteric types 40 and 41 that are considered as the second most common agents of gastroenteritis in children next to rotaviruses. The here-employed 293A cell line used for infectious status assessment showed its ability to multiply adenoviruses including type 41. Two modifications were moreover applied to the workflow for viral detection. The first occurred at the nucleic acid extraction step performed directly on all infected cells, while the second was the application of real-time quantitative PCR as detection tool. All adaptations led to a 3-day reduction of the response delay and an improved sensitivity especially for the enteric adenoviral types. The infectious status of laboratory strain types 2 and 41 was demonstrated by a more than 2-log10 increase in genome quantity. These conclusions were confirmed and reinforced by the analysis of water samples applying the improved assay. Naturally occurring infectious adenoviruses were detected in wastewater and river water, within 2 days. Types belonging to the species human adenoviruses C and type 31 were observed, but the most frequently identified type was 41 (71 % of identified sequences, n = 34). This highlights the usefulness of our method for a wide range of types, and especially for the most prevalent and public health-relevant enteric adenoviruses.

Is plant biomass input driving soil organic matter formation processes in grassland soil under contrasting management?

Grassland management practices vary in stocking rates and plant removal strategies (grazing versus mowing). They influence organic matter (OM) inputs, which were postulated as main controls of soil organic carbon (SOC) sequestration and might therefore control SOC stabilization. The aim of this study was to test this hypothesis by investigating the impacts of grassland harvesting regimes on parameters related to soil microbial functioning and soil organic matter (SOM) formation processes. We used a thirteen-year experiment in Central France under contrasting management (unmanaged, grazing with two intensities, mowing, bare fallow) to establish a carbon input gradient based on biomass leftovers after harvest. We investigated microbial biomass, basal respiration and enzyme activities as indicators of microbial functioning, and amino sugar content and composition as indicator of persistent SOM formation and origin through necromass accumulation. Responses of these parameters to carbon input along the gradient were contrasting and in most cases unrelated. Only the microbial C/N ratio and amino sugar contents showed a linear response indicating that they are influenced by plant-derived OM input. Other parameters were most probably more influenced by root activity, presence of herbivores, and/or physicochemical changes following management activities impacting soil microbial functioning. Grassland harvesting strategies influence SOC sequestration not only by changing carbon input quantity, but also through their effects on belowground processes possibly related to changing carbon input types and physiochemical soil properties.

Improved rock phosphate dissolution from organic acids is driven by nitrate assimilation of bacteria isolated from nitrate and CaCO3-rich soil.

Until now, the solubilization capacities of insoluble mineral P by soil microorganisms have been screened in vitro with media containing NH4+ as a nitrogen source. This presence of NH4+ will lead to an acidification of the medium responsible for the solubilization of the insoluble P. However, besides proton release, the production of organic acids can play a very important role in the release of free P. This physiological mechanism can largely depend on the source of nitrogen (NH4+vs NO3-) assimilated by the bacteria but the influence of the N source on the production of organic acids has yet to be studied. Our aim was to investigate if the N source assimilated by bacteria and the soil characteristics such as the dominant N source (NH4+vs NO3-) and CaCO3 contents might influence the bacterial capacities to solubilize rock phosphate. To fill this objective, we screened the capacity of bacteria isolated from 3 soils to solubilize rock phosphate in vitro in presence of NH4+or NO3-. Then, we selected the most efficient bacterial strains to identify and quantify the release of organic anions into the medium. Among the two hundred and forty-three bacterial strains isolated from the 3 soils, nine and seven isolates were identified with the highest % rock phosphate-solubilization values with NH4+ or NO3- as the sole N-source. Only one strain was able to release free Pi with NH4+ or NO3- as the sole N-source. The most predominant organic acids released by almost all isolates were gluconic acid, lactic acid, glycolic acid, acetic acid, formic acid and pyruvic acid regardless the N-source. However, with NO3- as source of N, the highest concentrations on those acids were found together with the highest release of free Pi into the medium. Molecular analysis of 16S rRNA indicated that almost all strains belonged to Bacillus and Paenibacillus genera. The PCA analysis between soil properties and bacterial capacities to release organic acids and free Pi also revealed that soil factors such as CaCO3 and soil NO3- content positively influenced the release of organic acids by bacteria grown in vitro. Our results concluded that the bacterial rock phosphate-solubilization was intimately related to organic acids production which in turn seemed to be driven by the assimilation of NO3- by bacteria. Therefore, the N-source might be considered a key factor to take into consideration during the screening and selection of suitable strains involved in the P-solubilization.

Toward better monitoring of human noroviruses and F-specific RNA bacteriophages in aquatic environments using bivalve mollusks and passive samplers: A case study.

Monitoring pathogenic enteric viruses in continental and marine water bodies is essential to control the viral contamination of human populations. Human Noroviruses (NoV) are the main enteric viruses present in surface waters and foodstuff. In a context of global change, it is currently a challenge to improve the management of viral pollutions in aquatic environments and thereby limit the contamination of vulnerable water bodies or foodstuffs. The aim of this study is to evaluate the potential of specific accumulation systems for improving the detection of NoV in water bodies, compared to direct water analyses. Passive samplers (Zetapor filters) and three species of bivalve molluscan shellfish (BMS) (Dreissena polymorpha, Mytilus edulis and Crassostreas gigas) were used as accumulation systems to determine their performance in monitoring continental and marine waters for viruses. F-specific RNA bacteriophages (FRNAPH) were also analyzed since they are described as indicators of NoV hazard in many studies. During a one-year study in a specific area frequently affected by fecal pollution, twelve campaigns of exposure of passive samplers and BMS in continental and coastal waters were conducted. Using suitable methods, NoV (genome) and FRNAPH (infectious and genome) were detected in these accumulation systems and in water at the same time points to determine the frequency of detection but also to gain a better understanding of viral pollution in this area. The reliability of FRNAPH as a NoV indicator was also investigated. Our results clearly showed that BMS were significantly better than passive samplers and direct water analyses for monitoring NoV and FRNAPH contamination in water bodies. A dilution of viral pollution between the continental and the coastal area was observed and can be explained by the distance from the source of the pollution. Viral pollution is clearly greater during the winter period, and stakeholders should take this into consideration in their attempts to limit the contamination of food and water. A significant correlation was once again shown between NoV and FRNAPH genomes in BMS, confirming the reliability of FRNAPH as a NoV indicator. Moreover, a strong correlation was observed between NoV genomes and infectious FRNAPH, suggesting recent viral pollution since infectious particles had not been inactivated at sufficient levels in the environment. More generally, this study shows the value of using BMS as an active method for improving knowledge on the behavior of viral contamination in water bodies, the ranking of the contamination sources, and the vulnerability of downstream water bodies.

SARS-CoV-2 Variability in Patients and Wastewaters-Potential Immuno-Modulation during the Shift from Delta to Omicron.

The continuous emergence of SARS-CoV-2 variants favors potential co-infections and/or viral mutation events, leading to possible new biological properties. The aim of this work was to characterize SARS-CoV-2 genetic variability during the Delta-Omicron shift in patients and in a neighboring wastewater treatment plant (WWTP) in the same urban area. The surveillance of SARS-CoV-2 was performed by routine screening of positive samples by single nucleotide polymorphism analysis within the S gene. Moreover, additionally to national systematic whole genome sequencing (WGS) once a week in SARS-CoV-2-positive patients, WGS was also applied when mutational profiles were difficult to interpret by routine screening. Thus, WGS was performed on 414 respiratory samples and on four wastewater samples, northeastern France. This allowed us to report (i) the temporally concordant Delta to Omicron viral shift in patients and wastewaters; (ii) the characterization of 21J (Delta) and 21K (Omicron)/BA.1-21L (Omicron)/BA.2-BA.4 mixtures from humans or environmental samples; (iii) the mapping of composite mutations and the predicted impact on immune properties in the viral Spike protein.