A review of organophosphonates, their natural and anthropogenic sources, environmental fate and impact on microbial greenhouse gases emissions - Identifying knowledge gaps.

Organophosphonates (OPs) are a unique group of natural and synthetic compounds, characterised by the presence of a stable, hard-to-cleave bond between the carbon and phosphorus atoms. OPs exhibit high resistance to abiotic degradation, excellent chelating properties and high biological activity. Despite the huge and increasing scale of OP production and use worldwide, little is known about their transportation and fate in the environment. Available data are dominated by information concerning the most recognised organophosphonate - the herbicide glyphosate - while other OPs have received little attention. In this paper, a comprehensive review of the current state of knowledge about natural and artificial OPs is presented (including glyphosate). Based on the available literature, a number of knowledge gaps have been identified that need to be filled in order to understand the environmental effects of these abundant compounds. Special attention has been given to GHG-related processes, with a particular focus on CH4. This stems from the recent discovery of OP-dependent CH4 production in aqueous environments under aerobic conditions. The process has changed the perception of the biogeochemical cycle of CH4, since it was previously thought that biological methane formation was only possible under anaerobic conditions. However, there is a lack of knowledge on whether OP-associated methane is also formed in soils. Moreover, it remains unclear whether anthropogenic OPs affect the CH4 cycle, a concern of significant importance in the context of the increasing rate of global warming. The literature examined in this review also calls for additional research into the date of OPs in waste and sewage and in their impact on environmental microbiomes.

Dynamics of Methane-Consuming Biomes from Wieliczka Formation: Environmental and Enrichment Studies.

The rocks surrounding Wieliczka salt deposits are an extreme, deep subsurface ecosystem that as we studied previously harbors many microorganisms, including methanotrophs. In the presented research bacterial community structure of the Wieliczka Salt Mine was determined as well as the methanotrophic activity of the natural microbiome. Finally, an enrichment culture of methane-consuming methanotrophs was obtained. The research material used in this study consisted of rocks surrounding salt deposits in the Wieliczka Salt Mine. DNA was extracted directly from the pristine rock material, as well as from rocks incubated in an atmosphere containing methane and mineral medium, and from a methanotrophic enrichment culture from this ecosystem. As a result, the study describes the composition of the microbiome in the rocks surrounding the salt deposits, while also explaining how biodiversity changes during the enrichment culture of the methanotrophic bacterial community. The contribution of methanotrophic bacteria ranged from 2.614% in the environmental sample to 64.696% in the bacterial culture. The methanotrophic enrichment culture was predominantly composed of methanotrophs from the genera Methylomonas (48.848%) and Methylomicrobium (15.636%) with methane oxidation rates from 3.353 ± 0.105 to 4.200 ± 0.505 µmol CH4 mL-1 day-1.

Application of nitrogen-rich sunflower husks biochar promotes methane oxidation and increases abundance of Methylobacter in nitrogen-poor soil.

The area of sunflower crops is steadily increasing. A beneficial way of managing sunflower waste biomass could be its use as a feedstock for biochar production. Biochar is currently being considered as an additive for improving soil parameters, including the ability to oxidise methane (CH4) - one of the key greenhouse gases (GHG). Despite the high production of sunflower husk, there is still insufficient information on the impact of sunflower husk biochar on the soil environment, especially on the methanotrophy process. To fill this knowledge gap, an experiment was designed to evaluate the effects of addition of sunflower husk biochar (produced at 450-550 °C) at a wide range of doses (1-100 Mg ha-1) to Haplic Luvisol. In the presented study, the CH4 oxidation potential of soil with and without sunflower husk biochar was investigated at 60 and 100% water holding capacity (WHC), and with the addition of 1% CH4 (v/v). The comprehensive study included GHG exchange (CH4 and CO2), physicochemical properties of soil (pH, soil organic carbon (SOC), dissolved organic carbon (DOC), nitrate nitrogen (NO3--N), WHC), and the structure of soil microbial communities. That study showed that even low biochar doses (5 and 10 Mg ha-1) were sufficient to enhance pH, SOC, DOC and NO3--N content. Importantly, sunflower husk biochar was significant source of NO3--N, which soil concentration increased from 9.40 ± 0.09 mg NO3--N kg-1 for the control to even 19.40 ± 0.26 mg NO3--N kg-1 (for 100 Mg ha-1). Significant improvement of WHC (by 11.0-12.4%) was observed after biochar addition at doses of 60 Mg ha-1 and higher. At 60% WHC, application of biochar at a dose of 40 Mg ha-1 brought significant improvements in CH4 oxidation rate, which was 4.89 ± 0.37 mg CH4-C kg-1 d-1. Higher biochar doses were correlated with further improvement of CH4 oxidation rates, which at 100 Mg ha-1 was seventeen-fold higher (8.36 ± 0.84 mg CH4-C kg-1 d-1) than in the biochar-free control (0.48 ± 0.28 mg CH4-C kg-1 d-1). CO2 emissions were not proportional to biochar doses and only grew circa (ca.) twofold from 3.16 to 6.90 mg CO2-C kg-1 d-1 at 100 Mg ha-1. Above 60 Mg ha-1, the diversity of methanotrophic communities increased, with Methylobacter becoming the most abundant genus, which was as high as 7.45%. This is the first, such advanced and multifaceted study of the wide range of sunflower husk biochar doses on Haplic Luvisol. The positive correlation between soil conditions, methanotroph abundance and CH4 oxidation confirmed the multifaceted, positive effect of sunflower husk biochar on Haplic Luvisol. Sunflower husk biochar can be successfully used for Haplic Luvisol supplementation. This additive facilitates soil protection against degradation and has the potential to mitigate GHG emissions.

Biochar dose determines methane uptake and methanotroph abundance in Haplic Luvisol.

Biochar promotes C sequestration and improvement of soil properties. Nevertheless, the effects of biochar addition on soil condition are poorly understood, especially with respect to greenhouse gas (GHG) emissions. A large proportion of GHG emissions derive from agriculture and, thus, recognition of the effect of biochar addition to soil on GHG emissions from terrestrial ecosystems is an important issue. The purpose of our study was to evaluate the short- and long-term effects of biochar application on soil in aspects of: GHG exchange (CH4 and CO2), basic physicochemical soil properties and structure of microbial communities in Haplic Luvisol. Soil was collected from fallow fields enriched with three doses of wood offcuts biochar (10, 20 and 30 Mg ha-1) and incubated at two moisture levels (60 and 100% WHC) with the addition of 1% CH4. To evaluate the influence of biochar aging in soil, the samples were analysed directly (short-term response) and five years (long-term response) after amendment. Generally, biochar addition increased soil pH, redox potential (Eh), organic carbon (SOC) and dissolved organic carbon (DOC) contents. Under 60% WHC, direct biochar application to the soil resulted in a clear improvement in the CH4 uptake rate. In contrast to that (at 100% WHC) methane uptake rates were twofold decreased. The positive effect was reduced due to biochar aging in the soil, but five years after application, at 60% WHC and the highest biochar dose (30 Mg ha-1) still significantly enhanced CH4 oxidation. From a short-term perspective, biochar application increased CO2 emissions, but after five years this effect was not observed. Microbial tests confirmed that the improvement in CH4 oxidation was correlated with methanotroph abundance in the soil. Moreover, an increase of Methylocystis abundance in the soil enriched with biochar along with enhanced CH4 uptake rates confirm the positive biochar influence on methanotrophic communities.

Biodegradation of Different Types of Plastics by Tenebrio molitor Insect.

Looking for new, sustainable ways to utilize plastics is still a very pertinent topic considering the amount of plastics produced in the world. One of the newest and intriguing possibility is the use of insects in biodegradation of plastics, which can be named entomoremediation. The aim of this work was to demonstrate the ability of the insect Tenebrio molitor to biodegrade different, real plastic waste. The types of plastic waste used were: remains of thermal building insulation polystyrene foam (PS), two types of polyurethane (kitchen sponge as PU1 and commercial thermal insulation foam as PU2), and polyethylene foam (PE), which has been used as packaging material. After 58 days, the efficiency of mass reduction for all of the investigated plastics was 46.5%, 41.0%, 53.2%, and 69.7% for PS, PU1, PU2, and PE, respectively (with a dose of 0.0052 g of each plastic per 1 mealworm larvae). Both larvae and imago were active plastic eaters. However, in order to shorten the duration of the experiment and increase the specific consumption rate, the two forms of the insect should not be combined together in one container.

A survey of greenhouse gases production in central European lignites.

Due to changes in the energy market, it is projected that lignite excavation will be reduced in the near future. Cessation of exploitation is associated with restitution of natural water conditions and flooding of the resources left in the mines. Flooded lignite mines are a potential source of greenhouse gases (GHG) (CH4, CO2 and N2O), which should be monitored due to growing environmental concerns. Here, we aim to recognize GHG release from the lignites collected from the main deposits of Poland, Slovenia and Serbia. GHG production was studied along with a range of physical and chemical parameters that are crucial for microbial growth and activity. The microcosm experiments showed that the main gas emitted from the lignites was carbon dioxide. Daily CO2 production was highly variable. The highest values were recorded for detroxylitic lignite collected from the Konin deposit (402.05 nmol CO2 g-1 day-1) while the lowest were for the Kolubara xylitic lignite (19.64 nmol CO2 g-1 day-1). Methane production was much lower and ranged from nearly zero to 66.75 nmol g dry mass-1 d-1. Nitrous oxide production was not detected. It was found that CO2 production, being a general measure of microbial activity, was positively affected by NO3- concentration and redox potential. With respect to methane formation, the lower atmospheric oxygen exposure of the sample from the Velenje underground mine compared to the samples from the opencast mines has been identified as a possible cause of the high methane production. The overall global warming potential (GWP) of the gases released by xylitic lignite was lowest among the samples. Preferential extraction of the detritic lignites is suggested as a means to reduce GHG emissions from the abandoned lignite mines.

Microbial Involvement in Carbon Transformation via CH4 and CO2 in Saline Sedimentary Pool.

Methane and carbon dioxide are one of the most important greenhouse gases and significant components of the carbon cycle. Biogeochemical methane transformation may occur even in the extreme conditions of deep subsurface ecosystems. This study presents methane-related biological processes in saline sediments of the Miocene Wieliczka Formation, Poland. Rock samples (W2, W3, and W4) differed in lithology (clayey salt with veins of fibrous salt and lenses of gypsum and anhydrite; siltstone and sandstone; siltstone with veins of fibrous salt and lenses of anhydrite) and the accompanying salt type (spiza salts or green salt). Microbial communities present in the Miocene strata were studied using activity measurements and high throughput sequencing. Biological activity (i.e., carbon dioxide and methane production or methane oxidation) occurred in all of the studied clayey salt and siltstone samples but mainly under water-saturated conditions. Microcosm studies performed at elevated moisture created more convenient conditions for the activity of both methanogenic and methanotrophic microorganisms than the intact sediments. This points to the fact that water activity is an important factor regulating microbial activity in saline subsurface sediments. Generally, respiration was higher in anaerobic conditions and ranged from 36 ± 2 (W2200%t.w.c) to 48 ± 4 (W3200%t.w.c) nmol CO2 gdw-1 day-1. Methanogenic activity was the highest in siltstone and sandstone (W3, 0.025 ± 0.018 nmol CH4 gdw-1 day-1), while aerobic methanotrophic activity was the highest in siltstone with salt and anhydrite (W4, 220 ± 66 nmol CH4 gdw-1 day-1). The relative abundance of CH4-utilizing microorganisms (Methylomicrobium, Methylomonas, Methylocystis) constituted 0.7-3.6% of all taxa. Methanogens were represented by Methanobacterium (0.01-0.5%). The methane-related microbes were accompanied by a significant number of unclassified microorganisms (3-64%) and those of the Bacillus genus (4.5-91%). The stable isotope composition of the CO2 and CH4 trapped in the sediments suggests that methane oxidation could have influenced δ13CCH4, especially in W3 and W4.

Influence of pipe material on biofilm microbial communities found in drinking water supply system.

The biofilms and water samples from a model installation built of PVC-U, PE-HD and cast iron pipes were investigated using standard heterotrophic plate count and 16S rRNA Next Generation Sequencing. The results of the high throughput identification imply that the construction material strongly influences the microbiome composition. PVC-U and PE-HD pipes were dominated with Proteobacteria (54-60%) while the cast pipe was overgrown by Nitrospirae (64%). It was deduced that the plastic pipes create a more convenient environment for the potentially pathogenic taxa than the cast iron. The 7-year old biofilms were described as complex habitats with sharp oxidation-reduction gradients, where co-existence of methanogenic and methanotrophic microbiota takes place. Furthermore, it was found that the drinking water distribution systems (DWDS) are a useful tool for studying the ecology of rare bacterial phyla. New ecophysiological aspects were described for Aquihabitans, Thermogutta and Vampirovibrio. The discrepancy between identity of HPC-derived bacteria and NGS-revealed composition of biofilm and water microbiomes point to the need of introducing new diagnostical protocols to enable proper assessment of the drinking water safety, especially in DWDSs operating without disinfection.

Biochar addition reinforces microbial interspecies cooperation in methanation of sugar beet waste (pulp).

Biogas production and microbial community structure were analyzed as an effect of biochar addition to a fermentation sludge containing sugar beet pulp. Positive effects of the treatment including an increase in process efficiency and better biogas quality were noted. The effect of biochar on AD (anaerobic digestion process) microbial communities was investigated after total DNA extraction from biochar-amended fermentation mixtures by PCR amplification of bacterial 16S rRNA gene fragments and Illumina amplicon sequencing. A combination of microbiological and physico-chemical analyses was used to study the mechanism by which biochar influences the process of anaerobic digestion of sugar beep pulp. It was found that the main reason of the changes in biogas production was the reshaping of the microbial communities, in particular enrichment of Bacteroidales and Clostridiales. It was proposed that biochar, in addition to being a conductor for mediating interspecies electron transfer, serves also as a habitat for hydrolytic bacteria. It was elucidated that the main driving force for the preferential colonization of biochar surfaces is its hydrophobicity. The presented research indicates the high potential of biochar to stimulate the methane fermentation process.

Changes in the Substrate Source Reveal Novel Interactions in the Sediment-Derived Methanogenic Microbial Community.

Methanogenesis occurs in many natural environments and is used in biotechnology for biogas production. The efficiency of methane production depends on the microbiome structure that determines interspecies electron transfer. In this research, the microbial community retrieved from mining subsidence reservoir sediment was used to establish enrichment cultures on media containing different carbon sources (tryptone, yeast extract, acetate, CO2/H2). The microbiome composition and methane production rate of the cultures were screened as a function of the substrate and transition stage. The relationships between the microorganisms involved in methane formation were the major focus of this study. Methanogenic consortia were identified by next generation sequencing (NGS) and functional genes connected with organic matter transformation were predicted using the PICRUSt approach and annotated in the KEGG. The methane production rate (exceeding 12.8 mg CH4 L-1 d-1) was highest in the culture grown with tryptone, yeast extract, and CO2/H2. The analysis of communities that developed on various carbon sources casts new light on the ecophysiology of the recently described bacterial phylum Caldiserica and methanogenic Archaea representing the genera Methanomassiliicoccus and Methanothrix. Furthermore, it is hypothesized that representatives of Caldiserica may support hydrogenotrophic methanogenesis.