Antiviral susceptibility of clade 2.3.4.4b highly pathogenic avian influenza A(H5N1) viruses isolated from birds and mammals in the United States, 2022

Clade 2.3.4.4b highly pathogenic avian influenza (HPAI) A(H5N1) viruses that are responsible for devastating outbreaks in birds and mammals pose a potential threat to public health. Here, we evaluated their susceptibility to influenza antivirals. Of 1,015 sequences of HPAI A(H5N1) viruses collected in the United States during 2022, eight viruses (∼0.8%) had a molecular marker of drug resistance to an FDA-approved antiviral: three adamantane-resistant (M2-V27A), four oseltamivir-resistant (NA-H275Y), and one baloxavir-resistant (PA-I38T). Additionally, 31 viruses contained mutations that may reduce susceptibility to inhibitors of neuraminidase (NA) (n = 20) or cap-dependent endonuclease (CEN) (n = 11). A panel of 22 representative viruses was tested phenotypically. Overall, clade 2.3.4.4b A(H5N1) viruses lacking recognized resistance mutations were susceptible to FDA-approved antivirals. Oseltamivir was least potent at inhibiting NA activity, while the investigational NA inhibitor AV5080 was most potent, including against NA mutants. A novel NA substitution T438N conferred 12-fold reduced inhibition by zanamivir, and in combination with the known marker N295S, synergistically affected susceptibility to all five NA inhibitors. In cell culture-based assays HINT and IRINA, the PA-I38T virus displayed 75- to 108-fold and 37- to 78-fold reduced susceptibility to CEN inhibitors, baloxavir and the investigational AV5116, respectively. Viruses with PA-I38M or PA-A37T showed 5- to 10-fold reduced susceptibilities. As HPAI A(H5N1) viruses continue to circulate and evolve, close monitoring of drug susceptibility is needed for risk assessment and to inform decisions regarding antiviral stockpiling

Technosols on mining wastes in the subarctic: Efficiency of remediation under Cu-Ni atmospheric pollution

The copper-nickel factory's emissions in the Murmansk region, Russia, led to the degradation of plant cover and topsoil with the subsequent formation of industrial barrens. In this study, the industrial barrens were remediated by means of Technosol engineering, when grasses were sown on the two different types of mining wastes (carbonatite and serpentinite-magnesite) covered by hydroponic vermiculite. The serpentinite-magnesite waste was significantly different from the carbonatite waste in the content of silicon (Si) and manganese (Mn), pH, and texture. Both wastes had an alkaline pH level and high content of calcium (Ca) and magnesium (Mg). The vegetation and Technosol properties at the remediated sites were analyzed in 2017 and compared to the initial state (2010 year) to assess the efficiency of the long-term remediation. The quality and sustainability of Technosols based on the serpentinite-magnesite wastes were substantially higher compared to the carbonatite-based Technosol. Biomass and a projective cover of the grass community depended on Si content in the original mining waste and were found to be higher in the serpentinite-magnesite Technosol. The content of organic carbon and its fractions, microbial biomass and basal respiration after seven years of Technosol evolution was comparable to natural values. These parameters were directly related to plant cover state and were inversely proportional to copper (Cu) content in Technosol. The Technosol development led to the reduction of nickel (Ni) and Cu migration in soil-plant ecosystems due to neutralization and adsorption properties of mining wastes and phytostabilization by underground parts of grass communities. The Technosol development in its early stage of pedogenesis indicates the efficiency of applied remediation technology to the degraded acidic soil under the conditions of industrial atmospheric pollution

Carbon dioxide emission and soil microbial respiration activity of Chernozems under anthropogenic transformation of terrestrial ecosystems

The total soil CO2 emission (EM) and portion of microbial respiration were measured (in situ; May, June, July 2015) in Chernozems typical of virgin steppe, oak forest, bare fallow and urban ecosystems (Kursk region, Russia). In soil samples (upper 10 cm layer), the soil microbial biomass carbon (Cmic), basal respiration (BR) and fungi-to-bacteria ratio were determined and the specific microbial respiration (BR / Cmic = qCO2) was calculated. The EM was varied from 2.0 (fallow) to 23.2 (steppe) g СО2 m-2 d-1. The portion of microbial respiration in EM was reached in average 83, 51 and 60% for forest, steppe and urban, respectively. The soil Cmic and BR were decreased along a gradient of ecosystems transformation (by 4 and 2 times less, respectively), while the qCO2 of urban soil was higher (in average by 42%) compared to steppe, forest and fallow. In urban soil the Cmic portion in soil Сorg and Сfungi-to-Сorg ratio were by 2.6 and 2.4 times less than those for steppe. The relationship between microbial respiration and BR values in Chernozems of various ecosystems was significant (R2 = 0.57)

Organic matter accumulation by alkaline-constructed soils in heavily metal-polluted area of Subarctic zone

Purpose: The research aimed to investigate properties and functions of soils constructed from alkaline mining wastes of different origin to remediate the industrial barren resulted from long-term emissions of the copper-nickel factory in the Subarctic region (Kola Peninsula, Russia). Conventional indicators of the remediation effectiveness (pH and metal content in geochemical fractions) were related to the indicators of soil functions such as biomass production, accumulation of organic carbon, microbial activity, and soil respiration. Materials and methods: The experimental area included two sites with polluted and degraded Podzol and Histosol soils located in 1.5 and 0.7 km from the nonferrous (Cu-Ni) smelter, respectively. At the sites, artificial soil constructions were made from mining wastes or quarry sand covered by the vermiculite layer with lawn grasses planted on top. Plant biomass was collected every year starting from the experiment set-up. In 5 to 8 years, soil samples were collected on the layer basis, and chemical, biological, and morphological properties were analyzed. Sequential fractionation of metals was conducted using a modified Tessier’s scheme. The microbial biomass and its respiration activity were determined. Micromorphological studies were conducted using an optical microscope. Soil respiration was measured on-site by IRGA with simultaneous observations of soil moisture and temperature. Results: The plant growth and residues' deposition at both experimental sites triggered carbon accumulation and resulted in 2–3 times higher content of organic carbon in the upper constructed soil layer compared to the initial content in mining wastes. Carbon accumulation was a key driver for the development of soil microbial communities and had a positive effect on the metal immobilization. This effect was strengthened by high pH inherited from the alkaline wastes and resulted in the performance of constructed soils as geochemical barriers. In their upper layers, where the root biomass was the highest, about 30–60% of Cu and Ni were bound by organic matter. In the underlying polluted soil, the most toxic water-soluble metal fraction was completely neutralized; and the metal concentrations in exchangeable fraction decreased by a factor of four improving the habitat conditions of the microbiome. Organic matter accumulation by clay material with the formation of organo-mineral films was found in the vermiculite-lizardite variant. Conclusion: Soil constructions made from alkaline mining wastes in the Subarctic supported the development of plant and microbial communities, organic matter accumulation, and metal immobilization. This technology allows protecting the environment from further pollution under the continuous emissions of the copper-nickel factory

Consequences of Grazing Cessation for Soil Environment and Vegetation in a Subalpine Grassland Ecosystem

Areas covered by seminatural grasslands have been in constant decline for decades in Europe. This trend is particularly strong for mountain territories, where such traditional agricultural practices as cattle grazing are no longer economically feasible. This study was conducted in the subalpine pasture of Cinte Tesino (TN, Italy), where local farmers have applied the following different management strategies: shorter and longer grazing durations during the season and a complete abandonment for the last 15 years. We aimed to study how these different management strategies impact the functioning and diversity of vegetation and the chemical and biological characteristics of the soil. Species richness was higher in plots subjected to longer grazing with a prevalence of D. caespitosa in terms of biomass share. A decline in species richness in abandoned plots was accompanied by an increase in the share of other graminoids in collected biomass. A concomitant increase in leaf N concentration and light availability in grazed plots resulted in higher photosynthetic efficiency in some species, as revealed by the δ13C of plant tissues. Soils under grazing were characterised by a higher concentration of total and extractable N, almost doubled microbial biomass C and increased extracellular enzymes activity, evidencing nutrient cycling mobilization. While the microbial pool was characterised by lower mineralization rates, C was lost from the soil with 15 years of abandonment. The longer grazing season demonstrated to be the most beneficial, promoting species richness, C accumulation and better soil microbial functioning. A change in soil pH from strongly acidic to moderately acidic with longer grazing is likely one of the important factors adding to the success in the functioning of primary producers and decomposers in this site

Microscopic fungi in big cities : Biodiversity, source, and relation to pollution by potentially toxic metals

For the first time, a mycological analysis of outdoor urban environment (air, leaves, sealed surfaces) was carried in the cities of subarctic (Murmansk) and temperate (Moscow) climatic zones. The chemical composition of dust deposited on leaves of dominant tree species was taken as an indicator of the air quality. Assessment of the complex impact of factors (climate zone, type of substrate, anthropogenic load) on the quantitative and qualitative parameters of mycobiome was performed. Compared to Moscow, Murmansk was characterized by an increased number and concentrations of pollutants in the deposited dust. The number of culturable airborne fungi in Murmansk was substantially lower than in Moscow. Half of the species belonged to the opportunistic in both cities. Most dangerous opportunistic fungi were absent in the air of recreational zones but present on leaves surface and in road dust in all assessed zones of the cities. Dust chemical composition affected the diversity of fungi species. While the relationship of biological parameters with concentration of potentially toxic metals was generally negative, Cd increased the fraction of opportunistic fungi in road dust. The study revealed an importance of substrate in determining the sensitivity of outdoor mycobiome to pollution and highlighted its biological characteristics sensitive to climate

Diversity of Microbial Communities, PAHs, and Metals in Road and Leaf Dust of Functional Zones of Moscow and Murmansk

The impact of geographical factors, functional zoning, and biotope type on the diversity of microbial communities and chemical components in the dust of urban ecosystems was studied. Comprehensive analyses of bacterial and fungal communities, polycyclic aromatic hydrocarbons (PAHs), and metals in road and leaf dust in three urban zones of Murmansk and Moscow with contrasting anthropogenic load were conducted. We found that the structure of bacterial communities affected the functional zoning of the city, biotope type, and geographical components. Fungal communities were instead impacted only by biotope type. Our findings revealed that the structure of fungal communities was mostly impacted by PAHs whereas bacterial communities were sensitive to metals. Bacteria of the genus Sphingomonas in road and leaf dust as indicators of the ecological state of the urban ecosystems were proposed

Polycyclic Aromatic Hydrocarbon-Degrading Bacteria in Three Different Functional Zones of the Cities of Moscow and Murmansk

We performed a comparative study of the total bacterial communities and communities of cultivable polycyclic aromatic hydrocarbons (PAH)-degrading bacteria in different functional zones of Moscow and Murmansk that were formed under the influence of the PAH composition in road and leaf dust. The PAHs were determined by high-performance liquid chromatography (HPLC); the bacterial communities’ diversity was assessed by metabarcoding. The degraders were isolated by their direct plating on a medium with the PAHs. The PAH total quantity declined in the leaf dust from the traffic to the recreational zone. For the road dust, a negative gradient with pollution was observed for Rhodococcus and Acinetobacter degraders and for their relative abundance in the microbiome for the functional zones of Moscow. The opposite effect was observed in the Murmansk leaf dust for the Rothia and Pseudomonas degraders and in the Moscow road dust for Microbacterium. The PCA and linear regression analyses showed that the Micrococcus degraders in the dust were sensitive to anthropogenic pollution, so they can be used as a tool for monitoring anthropogenic changes in the biosphere. The data on the degraders’ and microbial communities’ diversity suggest that minor degrading strains can play a key role in PAH degradation

Phylloplane Biodiversity and Activity in the City at Different Distances from the Traffic Pollution Source

The phylloplane is an integrated part of green infrastructure which interacts with plant health. Taxonomic characterization of the phylloplane with the aim to link it to ecosystem functioning under anthropogenic pressure is not sufficient because only active microorganisms drive biochemical processes. Activity of the phylloplane remains largely overlooked. We aimed to study the interactions among the biological characteristics of the phylloplane: taxonomic diversity, functional diversity and activity, and the pollution grade. Leaves of Betula pendula were sampled in Moscow at increasing distances from the road. For determination of phylloplane activity and functional diversity, a MicroResp tool was utilized. Taxonomic diversity of the phylloplane was assessed with a combination of microorganism cultivation and molecular techniques. Increase of anthropogenic load resulted in higher microbial respiration and lower DNA amount, which could be viewed as relative inefficiency of phylloplane functioning in comparison to less contaminated areas. Taxonomic diversity declined with road vicinity, similar to the functional diversity pattern. The content of Zn in leaf dust better explained the variation in phylloplane activity and the amount of DNA. Functional diversity was linked to variation in nutrient content. The fraction of pathogenic fungi of the phylloplane was not correlated with any of the studied elements, while it was significantly high at the roadsides. The bacterial classes Gammaproteobacteria and Cytophagia, as well as the Dothideomycetes class of fungi, are exposed to the maximal effect of distance from the highway. This study demonstrated the sensitivity of the phylloplane to road vicinity, which combines the effects of contaminants (mainly Zn according to this study) and potential stressful air microclimatic conditions (e.g., low relative air humidity, high temperature, and UV level). Microbial activity and taxonomic diversity of the phylloplane could be considered as an additional tool for bioindication

Multilayer design of crN/MoN superhard protective coatings and their characterisation

Multilayer CrN/MoN transition metal nitride coatings were studied in this research. Films were deposited by vacuum arc deposition (Arc-PVD) from Cr and Mo cathodes in nitrogen atmosphere p = 0.4 Pa. Three series of samples with different values of negative bias voltage (−20, −150, and −300 V) applied to the surface were fabricated. Each series has samples with 11, 22, 44, 88, 180 and 354 layers while total thickness was maintained with the same value. Samples were studied by scanning electron microscopy (SEM) on cross-sections and coatings surface, energy-dispersive X-ray spectroscopy (EDS), electron backscatter diffraction (EBSD), high-resolution transmission electron microscopy (HRTEM), X-ray diffraction (XRD), micro-indentation. Two main cubic phases of γ-Mo N and cubic CrN were detected. It was observed that the crystal growth orientation changes while the negative bias voltage of the substrate decreases. The maximum values of hardness (38–42 GPa) among the studied samples were obtained for coatings with a minimal individual layer thickness of 20 nm deposited at U = −20 V. N 2 bFCT - Sumy State University(0116U002621). Foundation of Science and Technology (FCT) of Portugal [references NORTE-01- 0145-FEDER-022096, SFRH/BD/129614/2017], Network of Extreme Conditions Laboratories (NECL) and by Ukrainian state budget programs [No. 0116U006816, 0118U003579 and 0116U002621]. Partial support by COST Action CA15102 is also greatly appreciate