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Oil wastes produced in large amounts in the processes of oil extraction, refining, and transportation are of great environmental concern because of their mutagenicity, toxicity, high fire hazardousness, and hydrophobicity. About 40% of these wastes contain radionuclides; however, the effects of oil products and radionuclides on soil microorganisms are frequently studied separately. The effects on various microbial parameters of raw waste containing 575 g of total petroleum hydrocarbons (TPH) kg−1 waste, 4.4 of 226Ra, 2.8 of 232Th, and 1.3 kBq kg−1 of 40K and its treated variant (1.6 g kg−1 of TPH, 7.9 of 226Ra, 3.9 of 232Th, and 183 kBq kg−1 of 40K) were examined in a leaching column experiment to separate the effects of hydrocarbons from those of radioactive elements. The raw waste sample (H) was collected from tanks during cleaning and maintenance, and a treated waste sample (R) was obtained from equipment for oil waste treatment. Thermal steam treatment is used in the production yard to reduce the oil content. The disposal of H waste samples on the soil surface led to an increase in the TPH content in soil: it became 3.5, 2.8, and 2.2 times higher in the upper (0–20 cm), middle (20–40 cm), and lower (40–60cm) layers, respectively. Activity concentrations of 226Ra and 232Th increased in soil sampled from both H- and R- columns in comparison to their concentrations in control soil. The activity concentrations of these two elements in samples taken from the upper and middle layers were much higher for the R-column compared to the H-column, despite the fact that the amount of waste added to the columns was equalized with respect to the activity concentrations of radionuclides. The H waste containing both TPH and radionuclides affected the functioning of the soil microbial community, and the effect was more pronounced in the upper layer of the column. Metabolic quotient and cellulase activity were the most sensitive microbial parameters as their levels were changed 5–1.4 times in comparison to control ones. Changes in soil functional characteristics caused by the treated waste containing mainly radionuclides were not observed. PCR-SSCP (polymerase chain reaction – single strand conformation polymorphism) analysis followed by MDS (metric multidimensional scaling) and clustering analysis revealed that the shifts in microbial community structure were affected by both hydrocarbons and radioactivity. Thus, molecular methods permitted to reveal the effects on soil microbial community not only from hydrocarbons, which significantly altered functional characteristics of soil microbiome, but also from radioactive elements.

Protists, known as microeukaryotes, are a significant portion of soil microbial communities. They are crucial predators of bacteria and depend on bacterial community dynamics for the growth and evolution of protistan communities. In parallel, increased levels of atmospheric CO 2 significantly impact bacterial metabolic activity in rhizosphere soils. In this study, we investigated the effect of elevated atmospheric CO 2 levels on the metabolically active protist community composition and function and their co-occurrences with bacteria from bulk and rhizosphere soils from the Giessen Free-Air CO 2 enrichment grassland experiment. Metabarcoding sequencing data analyses of partial 18S rRNA from total soil RNA showed that elevated CO 2 concentrations stimulated only a few ASVs of phagotrophic predators of bacteria and other microeukaryotes, affecting protist community composition (P = 0.006, PERMANOVA). In parallel, phagotrophic and parasitic lineages appeared slightly favoured under ambient CO 2 conditions, results that were corroborated by microbial signature analyses. Cross-comparisons of protist-bacteria co-occurrences showed mostly negative relations between prokaryotes and microeukaryotes, indicating that the ongoing increase in atmospheric CO 2 will lead to changes in microbial soil communities and their interactions, potentially cascading to higher trophic levels in soil systems., (© 2024. The Author(s).)

Protists are important key players in the microbial loop and influence their environment by grazing, which leads to the return of nutrients into the soil and reduces pathogen pressure on plants. Specifically, protists on and around plant roots are important for plants' development and growth. For this study, the fourth most important crop in the world, Hordeum vulgare , was selected. Seeds of H. vulgare were inoculated with Acanthamoeba castellanii alone or with additional soil bacteria at the beginning and during the experiment. The germination of the seeds and the growth of the plants in pouches were monitored over 3 weeks. No differences were found in leaf growth, root growth, root and leaf nitrogen content or ammonia content of the liquid from the pouches. In contrast, the relative increase in root and leaf dry weight showed a small difference compared to the controls. The results of this experiment demonstrated that seed inoculation with A. castellanii alone or with additional unidentified soil bacteria did not have a major effect on the growth and development of barley. Nevertheless, small changes in plant development were detected, indicating that A. castellanii should be considered for further investigation of co-inoculations with plant growth-promoting bacteria and additional nutrients., Competing Interests: The authors declare that there are no conflicts of interest., (Copyright © 2024 The Authors.)

Modern crops might have lost some of their functional traits, required for interacting with beneficial microbes, as a result of the genotypic/phenotypic modifications that occurred during domestication. Here, we studied the bacterial and fungal microbiota in the rhizosphere of two cultivated wheat species (Triticum aestivum and T. durum) and their respective ancestors (Aegilops tauschii and T. dicoccoides), in three experimental fields, by using metabarcoding of 16S rRNA genes and ITS2, coupled with co-occurrence network analysis. Moreover, the abundance of bacterial genes involved in N- and P-cycles was estimated by quantitative PCR, and urease, alkaline phosphatase and phosphomonoesterase activities were assessed by enzymatic tests. The relationships between microbiota and environmental metadata were tested by correlation analysis. The assemblage of core microbiota was affected by both site and plant species. No significant differences in the abundance of potential fungal pathogens between wild and cultivated wheat species were found; however, co-occurrence analysis showed more bacterial-fungal negative correlations in the wild species. Concerning functions, the nitrogen denitrification nirS gene was consistently more abundant in the rhizosphere of A. tauschii than T. aestivum. Urease activity was higher in the rhizosphere of each wild wheat species in at least two of the research locations. Several microbiota members, including potentially beneficial taxa such as Lysobacter and new taxa such as Blastocatellaceae, were found to be strongly correlated to rhizospheric soil metadata. Our results showed that a functional microbiome shift occurred as a result of wheat domestication. Notably, these changes also included the reduction of the natural biocontrol potential of rhizosphere-associated bacteria against pathogenic fungi, suggesting that domestication disrupted the equilibrium of plant-microbe relationships that had been established during million years of co-evolution., (Copyright © 2024 The Authors. Published by Elsevier GmbH.. All rights reserved.)

Monitoring of bioinoculants once released into the field remains largely unexplored; thus, more information is required about their survival and interactions after root colonization. Therefore, specific primers were used to perform a long-term tracking to elucidate the effect of Hartmannibacter diazotrophicus on wheat and barley production at two experimental organic agriculture field stations. Three factors were evaluated: organic fertilizer application (with and without), row spacing (15 and 50 cm), and bacterial inoculation (H. diazotrophicus and control without bacteria). Hartmannibacter diazotrophicus was detected by quantitative polymerase chain reaction on the roots (up to 5 × 105 copies g-1 dry weight) until advanced developmental stages under field conditions during two seasons, and mostly in one farm. Correlation analysis showed a significant effect of H. diazotrophicus copy numbers on the yield parameters straw yield (increase of 453 kg ha-1 in wheat compared to the mean) and crude grain protein concentration (increase of 0.30% in wheat and 0.80% in barley compared to the mean). Our findings showed an apparently constant presence of H. diazotrophicus on both wheat and barley roots until 273 and 119 days after seeding, respectively, and its addition and concentration in the roots are associated with higher yields in one crop., (© The Author(s) 2024. Published by Oxford University Press on behalf of FEMS.)

The use of antibiotics unbalances the intestinal microbiota. Probiotics, prebiotics, and synbiotics are alternatives for these unbalances. The effects of a new synbiotic composed of probiotic Saccharomyces boulardii CNCM I-745 and fructans from Agave salmiana (fAs) as prebiotics were assessed to modulate the intestinal microbiota. Two probiotic presentations, the commercial probiotic (CP) and the microencapsulated probiotic (MP) to improve those effects, were used to prepare the synbiotics and feed Wistar rats subjected to antibiotics (AB). Eight groups were studied, including five controls and three groups to modulate the microbiota after the use of antibiotics: G5: AB + MP-synbiotic, G6: AB + CP-synbiotic, and G8: AB + fAs. All treatments were administered daily for 7 days. On days 7 and 21, euthanasia was performed, cecum tissue was recovered and used to evaluate histological analysis and to study microphotograph by TEM, and finally, bacterial DNA was extracted and 16S rRNA gene metabarcode sequencing was performed. Histological analysis showed less epithelial damage and more abundance of the intestinal microbiota in the groups G5, G6, and G8 in comparison with the AB control group after 7 days. Microphotograph of the cecum at 2 weeks post treatment showed that G5 and G6 presented beneficial effects in epithelial reconstruction. Interestingly, in the groups that used the synbiotic without AB (G3 and G4) in addition to contributing to the recovery of the autochthonous microbiota, it promotes the development of beneficial microorganisms; those results were also achieved in the groups that used the synbiotic with AB enhancing the bacterial diversity and regulating the impact of AB., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The obligate biotrophic fungal pathogen Blumeria graminis causes the powdery mildew disease of cereals, which results in large crop losses. Control of B. graminis in barley is mainly achieved by fungicide treatment and by breeding resistant varieties. Vampyrellid amoebae, just like mycophagous protists, are able to consume a variety of fungi. To reveal the impact of some selected fungus-consuming protists on Blumeria graminis f. sp. hordei (Bgh), and to evaluate the possibility of using these protists as biological agents in the future, their feeding behaviour on B. graminis spores on barley leaves was investigated. An experiment was carried out with five different protist isolates (Leptophrys vorax, Platyreta germanica, Theratromyxa weberi U 11, Theratromyxa weberi G7.2 and Acanthamoeba castellanii) and four matched controls, including the food sources of the cultures and the medium. Ten-day-old leaves of barley (Hordeum vulgare cv. Golden Promise) were first inoculated with Blumeria graminis (f. sp. hordei race A6) spores, then treated with protists and fungal colonies on the leaf surfaces were counted under the microscope after 5 days. The isolates L. vorax, P. germanica, and T. weberi U11 did not show a significant reduction in the number of powdery mildew colonies whereas the isolates T. weberi G7.2 and A. castellanii significantly reduced the number of powdery mildew colonies on the leaf surfaces compared to their respective controls. This indicates that these two isolates are capable of reducing B. graminis colonies on barley leaves and are suitable candidates for further investigation for possible use as biological agents. Nevertheless, the susceptibility to dryness and the cell division rate should be considered during the optimisation of the next steps like application procedure and whole plant treatment., (© 2023. The Author(s).)

One gram-negative strain designated Bb-Pol-6  T was isolated from birch (Betula pendula) pollen at Giessen area, Germany. The analysis of 16S rRNA gene-based phylogenies indicated the next-relative genera were Robbsia, Chitinasiproducens, Pararobbsia and Paraburkholderia (96-95.6%). Further comparative genome analysis and phylogenetic tree-based methods revealed its phylogenetic position under the genus Robbsia. The genome of strain Bb-Pol-6  T was 5.04 Mbp with 4401 predicted coding sequences and a G + C content of 65.31 mol%. Average amino acid identity, average nucleotide identity, digital DNA-DNA hybridization and percentage of conserved proteins values to Robbsia andropogonis DSM 9511  T were 68.0, 72.5, 22.7 and 65.85%, respectively. Strain Bb-Pol-6  T was rod-shaped, non-motile, facultative anaerobic and grew optimally at 28 °C and pH 6-7. Ubiquinone 8 was the major respiratory quinone and the major cellular fatty acids were C 16:0 , C 19:0 cyclo ω7c, C 17:0 cyclo ω7c and C 17:1 ω6c. The dominant polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and an unidentified aminophospholipid. Based on the genomic physiological and phenotypic characteristics, strain Bb-Pol-6  T was considered a novel species under the genus Robbsia, for which the name Robbsia betulipollinis sp. nov. was proposed. The type strain is Bb-Pol-6  T (= LMG 32774  T  = DSM 114812  T )., (© 2023. The Author(s).)

Dysbiosis has been implicated in childhood obesity. Oral intake of fermented milk containing Lacticaseibacillus casei strain Shirota preserves gut microbiota (GM) diversity in children and adults. This study was a double-blind trial involving 37 overweight or obese children aged 6-10 years. Children were followed over a 6-week intervention period in which they received different fermented milk products containing L. casei Shirota: 10 in the first group received just L. casei Shirota; 13 received L. casei Shirota with 3 g/day of inulin ( L. casei +inulin); and 14 received L. casei Shirota with 3 g/day of fructans from Agave salmiana ( L. casei +fructans). Principal component analysis showed the relationship between microbial abundance, GM metabolites, and other obesity-related markers. Supplementation with probiotics and synbiotics improved the HDL-cholesterol levels of overweight and obese children, although no changes in body composition were detected. We observed an increase in butyrate or propionate concentrations in the L. casei +fructans group compared to the end of the intervention ( P <0.03). A diminished level of ANGPTL4 within the L. casei +fructans group ( P =0.04) was also found, but no differences when lipopolysaccharide-binding protein was evaluated. The FFAR2+ cell frequency decreased between baseline and at the end of 6-week intervention in L. casei +inulin ( P =0.02) and L. casei +fructans groups ( P =0.04). In contrast, the percentage of CD14+FFAR3+ frequency increased in the same groups ( P =0.04). The L. casei Shirota with inulin or fructans modulates GM, which improves the lipid profile and changes at a molecular level, such as expression of FFAR3 and FFAR2, ANGPTL4, propionate, and butyrate. It, therefore, could be considered an interesting therapeutic possibility for treating childhood overweight and obesity. The study was registered at ClinicalTrials.gov (ID: NCT05423015).

Background: Elevated carbon dioxide concentrations (eCO 2 ), one of the main causes of climate change, have several consequences for both vine and cover crops in vineyards and potentially also for the soil microbiome. Hence soil samples were taken from a vineyard free-air CO 2 enrichment (VineyardFACE) study in Geisenheim and examined for possible changes in the soil active bacterial composition (cDNA of 16S rRNA) using a metabarcoding approach. Soil samples were taken from the areas between the rows of vines with and without cover cropping from plots exposed to either eCO 2 or ambient CO 2 (aCO 2 )., Results: Diversity indices and redundancy analysis (RDA) demonstrated that eCO 2 changed the active soil bacterial diversity in grapevine soil with cover crops (p-value 0.007). In contrast, the bacterial composition in bare soil was unaffected. In addition, the microbial soil respiration (p-values 0.04-0.003) and the ammonium concentration (p-value 0.003) were significantly different in the samples where cover crops were present and exposed to eCO 2 . Moreover, under eCO 2 conditions, qPCR results showed a significant decrease in 16S rRNA copy numbers and transcripts for enzymes involved in N 2 fixation and NO 2 - reduction were observed using qPCR. Co-occurrence analysis revealed a shift in the number, strength, and patterns of microbial interactions under eCO 2 conditions, mainly represented by a reduction in the number of interacting ASVs and the number of interactions., Conclusions: The results of this study demonstrate that eCO 2 concentrations changed the active soil bacterial composition, which could have future influence on both soil properties and wine quality., (© 2023. The Author(s).)

Sensitization to pollen allergens has been increasing in Europe every year. Most studies in this field are related to climate change, phenology, allergens associated with different pollens, and allergic disorders. As a plant microhabitat, pollen is colonized by diverse microorganisms, including endotoxin-producing bacteria which may contribute to pollen allergy (pollinosis). Therefore, bacteria isolated from high allergenic and low allergenic plant pollen, as well as the pollen itself with all microbial inhabitants, were used to assess the effect of the pollen by measuring the endotoxins lipopolysaccharides (LPS) and lipoteichoic acid (LTA) concentrations and their effect on chemokine and cytokine release from transwell cultured epithelial A549 cells as a model of epithelial lung barrier. High allergenic pollen showed a significantly higher level of bacterial endotoxins; interestingly, the endotoxin level found in the bacterial isolates from high allergenic pollen was significantly higher compared to that of bacteria from low allergenic pollen. Moreover, bacterial LPS concentrations across different pollen species positively correlated with the LPS concentration across their corresponding bacterial isolates. Selected bacterial isolates from hazel pollen (HA5, HA13, and HA7) co-cultured with A549 cells induced a potent concentration-dependent release of the chemokine interleukin-8 and monocyte chemotactic protein-1 as well as the cytokine TNF-alpha and interleukin-2 to both apical and basal compartments of the transwell model. This study clearly shows the role of bacteria and bacterial endotoxins in the pollen allergy as well as seasonal allergic rhinitis., (© 2022. The Author(s).)

Soil organisms play an important role in the equilibrium and cycling of nutrients. Because elevated CO 2 (eCO 2 ) affects plant metabolism, including rhizodeposition, it directly impacts the soil microbiome and microbial processes. Therefore, eCO 2 directly influences the cycling of different elements in terrestrial ecosystems. Hence, possible changes in the cycles of carbon (C), nitrogen (N), and sulfur (S) were analyzed, alongside the assessment of changes in the composition and structure of the soil microbiome through a functional metatranscriptomics approach (cDNA from mRNA) from soil samples taken at the Giessen free-air CO 2 enrichment (Gi-FACE) experiment. Results showed changes in the expression of C cycle genes under eCO 2 with an increase in the transcript abundance for carbohydrate and amino acid uptake, and degradation, alongside an increase in the transcript abundance for cellulose, chitin, and lignin degradation and prokaryotic carbon fixation. In addition, N cycle changes included a decrease in the transcript abundance of N 2 O reductase, involved in the last step of the denitrification process, which explains the increase of N 2 O emissions in the Gi-FACE. Also, a shift in nitrate ( NO 3 - ) metabolism occurred, with an increase in transcript abundance for the dissimilatory NO 3 - reduction to ammonium ( NH 4 + ) (DNRA) pathway. S metabolism showed increased transcripts for sulfate ( SO 4 2 - ) assimilation under eCO 2 conditions. Furthermore, soil bacteriome, mycobiome, and virome significantly differed between ambient and elevated CO 2 conditions. The results exhibited the effects of eCO 2 on the transcript abundance of C, N, and S cycles, and the soil microbiome. This finding showed a direct connection between eCO 2 and the increased greenhouse gas emission, as well as the importance of soil nutrient availability to maintain the balance of soil ecosystems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Rosado-Porto, Ratering, Moser, Deppe, Müller and Schnell.)

Elevated levels of atmospheric CO 2 lead to the increase of plant photosynthetic rates, carbon inputs into soil and root exudation. In this work, the effects of rising atmospheric CO 2 levels on the metabolic active soil microbiome have been investigated at the Giessen free-air CO 2 enrichment (Gi-FACE) experiment on a permanent grassland site near Giessen, Germany. The aim was to assess the effects of increased C supply into the soil, due to elevated CO 2 , on the active soil microbiome composition. RNA extraction and 16S rRNA (cDNA) metabarcoding sequencing were performed from bulk and rhizosphere soils, and the obtained data were processed for a compositional data analysis calculating diversity indices and differential abundance analyses. The structure of the metabolic active microbiome in the rhizospheric soil showed a clear separation between elevated and ambient CO 2 (p = 0.002); increased atmospheric CO 2 concentration exerted a significant influence on the microbiomes differentiation (p = 0.01). In contrast, elevated CO 2 had no major influence on the structure of the bulk soil microbiome (p = 0.097). Differential abundance results demonstrated that 42 bacterial genera were stimulated under elevated CO 2 . The RNA-based metabarcoding approach used in this research showed that the ongoing atmospheric CO 2 increase of climate change will significantly shift the microbiome structure in the rhizosphere., (© 2021. The Author(s).)

The seed-transmitted microorganisms and the microbiome of the soil in which the plant grows are major drivers of the rhizosphere microbiome, a crucial component of the plant holobiont. The seed-borne microbiome can be even coevolved with the host plant as a result of adaptation and vertical transmission over generations. The reduced genome diversity and crossing events during domestication might have influenced plant traits that are important for root colonization by seed-borne microbes and also rhizosphere recruitment of microbes from the bulk soil. However, the impact of the breeding on seed-transmitted microbiome composition and the plant ability of microbiome selection from the soil remain unknown. Here, we analyzed both endorhiza and rhizosphere microbiome of two couples of genetically related wild and cultivated wheat species ( Aegilops tauschii / Triticum aestivum and T. dicoccoides / T. durum ) grown in three locations, using 16S rRNA gene and ITS2 metabarcoding, to assess the relative contribution of seed-borne and soil-derived microbes to the assemblage of the rhizosphere microbiome. We found that more bacterial and fungal ASVs are transmitted from seed to the endosphere of all species compared with the rhizosphere, and these transmitted ASVs were species-specific regardless of location. Only in one location, more microbial seed transmission occurred also in the rhizosphere of A. tauschii compared with other species. Concerning soil-derived microbiome, the most distinct microbial genera occurred in the rhizosphere of A. tauschii compared with other species in all locations. The rhizosphere of genetically connected wheat species was enriched with similar taxa, differently between locations. Our results demonstrate that host plant criteria for soil bank's and seed-originated microbiome recruitment depend on both plants' genotype and availability of microorganisms in a particular environment. This study also provides indications of coevolution between the host plant and its associated microbiome resulting from the vertical transmission of seed-originated taxa., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Abdullaeva, Ratering, Ambika Manirajan, Rosado-Porto, Schnell and Cardinale.)

In this study, the taxonomic and functional diversity of methanogenic archaea in two parallel 120 l fermenters operated at different temperatures and fed with maize silage was estimated by mcrA metabarcoding analysis using two typical primer pairs (ML and MLA) amplifying part of the functional methyl coenzyme M reductase (mcrA) gene. The alpha diversity indices showed that the ML primer pair detected a higher Operational Taxonomic Unit (OTU) abundance compared to the MLA primer pair and methanogen diversity was significantly lower in the 60 °C fermenters. The beta diversity analysis showed the methanogenic community clustered together at 50 °C and 40° and was statistically different from the 60 °C community. Similar, to alpha diversity, beta diversity was also significantly different between primer pairs. At all temperatures analysed, the primer pairs showed a different abundance of the different methanogenic OTUs, e.g. more OTUs relative to Methanoculleus sp. with the ML primer pair, and more OTUs corresponding to Methanobacterium sp. with the MLA primer pair. Moreover, OTUs corresponding to Methanosphaera sp. and Methanobrevibacter sp. were found only by using ML primer pair, while the MLA primer pair detected sequences corresponding to Methanothrix sp., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

Microbially contaminated washing machines and mild laundering conditions facilitate the survival and growth of microorganisms on laundry, promoting undesired side effects such as malodor formation. Clearly, a deeper understanding of the functionality and hygienic relevance of the laundry microbiota necessitates the analysis of the microbial gene expression on textiles after washing, which-to the best of our knowledge-has not been performed before. In this pilot case study, we used single-end RNA sequencing to generate de novo transcriptomes of the bacterial communities remaining on polyester and cotton fabrics washed in a domestic washing machine in mild conditions and subsequently incubated under moist conditions for 72 h. Two common de novo transcriptome assemblers were used. The final assemblies included 22,321 Trinity isoforms and 12,600 Spades isoforms. A large part of these isoforms could be assigned to the SwissProt database, and was further categorized into "molecular function", "biological process" and "cellular component" using Gene Ontology (GO) terms. In addition, differential gene expression was used to show the difference in the pairwise comparison of the two tissue types. When comparing the assemblies generated with the two assemblers, the annotation results were relatively similar. However, there were clear differences between the de novo assemblies regarding differential gene expression.

A Gram-stain-negative bacterium, designated I-24 T , was isolated from soil of a natural salt meadow. Strain I-24 T was aerobic, non-motile, rod-shaped, catalase-positive, oxidase-positive and grew optimally at pH 7 and 25 °C. Comparative 16S rRNA gene analysis indicated that strain I-24 T has closest similarities to Spirosoma agri KCTC 52727 T (95.9 %) and Spirosoma terrae KCTC 52035 T (95.5 %). Strain I-24 T contained summed feature 3 (C 16 : 1  ω 7 c /C 16 : 1  ω 6 c ) and C 16 : 1  ω 5 c as the major fatty acids, the predominant respiratory quinone was menaquinone MK-7, and the major polar lipids were phosphatidylethanolamine as well as an unidentified phosphoaminolipid. The draft genome of strain I-24 T consists of 10 326 072 base pairs with 9153 predicted coding sequences and a G+C content of 47.7 mol%. Clear distinctions between strain I-24 T and S. agri KCTC 52727 T or S. terrae KCTC 52035 T were shown in the pairwise average nucleotide identity results with values of 76.71 and 74.01 %, respectively. Moreover, the digital DNA-DNA relatedness values to these strains were 20.8 and 19.0 %. Based on its phenotypic, genotypic and chemotaxonomic characteristics, strain I-24 T represents a novel species of the genus Spirosoma , for which the name Spirosoma endbachense sp. nov. is proposed. The type strain is I-24 T (DSM 111055 T =KCTC 72613 T ).

Spirosoma agri S7-3-3 (KCTC 52727) and Spirosoma terrae 15J9-4 (KCTC 52035) are type strains isolated from an apple orchard and beach soil in South Korea, respectively; their draft genome sequences were assembled and annotated. The draft genome sequences of S7-3-3 T (7,239,915 bp; G+C content, 50.6%) and 15J9-4 T (7,551,610 bp; G+C content, 47.3%) are reported., (Copyright © 2020 Rojas et al.)

Healthy Wistar rats were supplemented during 20 weeks with commercial inulin (I) and Agave tequilana fructans (CAT), experimental fructans from A. tequilana (EAT) and A. salmiana (AS) mature stems, rice starch 10% (RS), and standard feed for rodents (C). Feed intake was kept steady, but with I, body weight and abdominal adipose tissue (6.01 g) decreased at the end. Glucose (mg/dL) (C, 120.52; I, 110.69; CAT, 105.75; EAT, 115.48; AS, 101.63; and RS, 121.82), total cholesterol (C, 89.89; I, 64.48; CAT, 68.04; EAT, 68.74; AS, 68.04; and RS, 82), and triglycerides (C, 84.03; I, 59.52; CAT, 68.56; EAT, 59.08; AS, 75.27; and RS, 81.8) kept being normal and without differences between fructans. At the end, there was a significant increase in lactic acid bacteria when the I and AS groups were compared to the C group (C, 9.18; I, 10.64; CAT, 10.34; EAT, 10.36; AS, 10.49; and RS, 9.62 log 10 CFU/g of feces). In addition, with fructans, there was an accelerated process in feces emptiness, Lieberkühn crypts kept their morphology, and there was an increment of goblet cells., Competing Interests: The authors declare no competing financial interest., (Copyright © 2020 American Chemical Society.)

The effects of different agronomic practices, such as fertilization regimes, can be experimentally tested in long-term experiments (LTE). Here, we aimed to evaluate the effect of different nitrogen fertilizations on the bacterial microbiota in both rhizosphere and bulk soil of sugar beet, in the Giessen-LTE (Germany). Fertilization treatments included mineral-N, manure, mineral-N + manure and no N-amendment. Metabarcoding and co-occurrence analysis of 16S rRNA genes, qPCR of amoA , nirK , nirS , nosZ -I and nosZ -II genes and soil physico-chemical analyses were performed. The effect of the fertilization treatments was more evident in the bulk soil, involving 33.1% of the microbiota. Co-occurrence analysis showed a rhizosphere cluster, dominated by Proteobacteria, Actinobacteria and Verrucomicrobia (hub taxa: Betaproteobacteriales ) , and a bulk soil cluster, dominated by Acidobacteria , Gemmatominadetes and " Latescibacteria " (hub taxa: Acidobacteria ). In the bulk soil, mineral N-fertilization reduced nirK , amoA , nosZ -I and nosZ -II genes. Thirteen Operational taxonomic units (OTUs) showed 23 negative correlations with gene relative abundances. These OTUs likely represent opportunistic species that profited from the amended mineral-N and outgrew the species carrying N-cycle genes. Our results indicate trajectories for future research on soil microbiome in LTE and add new experimental evidence that will be helpful for sustainable management of nitrogen fertilizations on arable soils.

Modern, mainly sustainability-driven trends, such as low-temperature washing or bleach-free liquid detergents, facilitate microbial survival of the laundry processes. Favourable growth conditions like humidity, warmth and sufficient nutrients also contribute to microbial colonization of washing machines. Such colonization might lead to negatively perceived staining, corrosion of washing machine parts and surfaces, as well as machine and laundry malodour. In this study, we characterized the bacterial community of 13 domestic washing machines at four different sampling sites (detergent drawer, door seal, sump and fibres collected from the washing solution) using 16S rRNA gene pyrosequencing and statistically analysed associations with environmental and user-dependent factors. Across 50 investigated samples, the bacterial community turned out to be significantly site-dependent with the highest alpha diversity found inside the detergent drawer, followed by sump, textile fibres isolated from the washing solution, and door seal. Surprisingly, out of all other investigated factors only the monthly number of wash cycles at temperatures ≥ 60 °C showed a significant influence on the community structure. A higher number of hot wash cycles per month increased microbial diversity, especially inside the detergent drawer. Potential reasons and the hygienic relevance of this finding need to be assessed in future studies.

Pleurotus ostreatus is one of the most cultivated edible mushrooms worldwide and few approaches have been done to analyze bacterial influence during its cultivation. Therefore, bacteria from commercial spawn, mycelial-colonized straw and fruiting bodies from healthy productive samples were counted, isolated and tested for their mycelial growth promoting ability. Bacterial cell numbers at different steps of the process showed low bacterial cell numbers in spawn and in fruiting bodies inner tissue compared to the high concentration in mycelial-colonized straw. The majority of the 38 isolates belonged to phyla Firmicutes and Actinobacteria were identified as Bacillus, Paenibacillus and Micromonospora species. Similarly, 16S rRNA gene bacterial clones obtained from mycelial biomass DNA samples showed bacterial presence of various genera including Bacillus and Paenibacillus. In the mycelial growth promoting ability tests, 30 isolates negatively affected mycelial growth, two isolates showed no effect on mycelial growth, and six isolates promoted mycelial growth. Moreover, mycelial thickness was influenced in different ways by the bacterial growth. In general, nearly all isolates growth-preventing were isolated from healthy spawn and mycelial-colonized straw, whereas fruiting bodies were the best source for isolation of mycelial growth-promoting bacteria. Characterization of bacterial isolates revealed that growth-preventing isolates exhibited various enzymatic activities in comparison with positive influencing bacteria that exhibited none or weak enzymatic activities. In addition, the influence of volatile compounds being present in the headspace of bi-plate co-cultures on P. ostreatus mycelial growth was demonstrated. The effect of isolates, that promoted mycelial growth in co-cultivation, to reduce P. ostreatus spawn running time, was evaluated on sterilized rye seeds. Results showed that not all mycelial promoted isolates were able to significantly promote P. ostreatus colonization. However, isolate M46F identified as Micromonospora lupini significantly reduce spawn running time. This is one of few studies to estimate cultivable bacteria from healthy samples of P. ostreatus cultivation, to evaluate a bacterial effect on mycelial growth, to show that fruiting bodies are a good source for mycelia growth-promoting isolates, and the first to report a shorter P. ostreatus spawn running time due to bacterial inoculation., Competing Interests: Declaration of Competing Interest The authors declare no potential conflict of interest, (Copyright © 2020 Elsevier GmbH. All rights reserved.)

Parkinson's disease (PD) is one of the most common neurodegenerative disorders. PD patients suffer from gastrointestinal dysfunctions and alterations of the autonomous nervous system, especially its part in the gut wall, i.e., the enteric nervous system (ENS). Such alterations and functional gastrointestinal deficits often occur years before the classical clinical symptoms of PD appear. Until now, only little is known about PD-associated changes in gut microbiota composition and their potential implication in PD development. In order to increase knowledge in this field, fecal samples of 34 PD patients and 25 healthy, age-matched control persons were investigated. Here, the V4 and V5 hypervariable region of bacterial 16S rRNA genes was PCR-amplified and sequenced using an Ion Torrent PGM platform. Within the PD group, we observed a relative decrease in bacterial taxa which are linked to health-promoting, anti-inflammatory, neuroprotective or other beneficial effects on the epithelial barrier, such as Faecalibacterium and Fusicatenibacter . Both taxa were lowered in PD patients with elevated levels of the fecal inflammation marker calprotectin. In addition, we observed an increase in shares of the Clostridiales family XI and their affiliated members in these samples. Finally, we found that the relative abundances of the bacterial genera Peptoniphilus, Finegoldia, Faecalibacterium Fusicatenibacter, Anaerococcus, Bifidobacterium, Enterococcus , and Ruminococcus were significantly influenced by medication with L-dopa and entacapone, respectively. Our data confirm previously reported effects of COMT inhibitors on the fecal microbiota of PD patients and suggest a possible effect of L-dopa medication on the relative abundance of several bacterial genera., Competing Interests: Competing interestsThe authors declare that this research was conducted in the absence of commercial or financial relationships that could be construed as potential conflicts of interest., (© The Author(s) 2019.)

Strain E19 T described as Hartmannibacter diazotrophicus gen. nov. sp. nov. was isolated from the rhizosphere of Plantago winteri from a natural salt meadow in a nature protection area. Strain E19 T is a plant growth-promoting rhizobacterium able to colonize the rhizosphere of barley and to promote its growth only under salt stress conditions. To gain insights into the genetic bases of plant growth promotion and its lifestyle at the rhizosphere under salty conditions, we determined the complete genome sequence using two complementary sequencing platforms (Ilumina MiSeq and PacBio RSII ). The E19 T genome comprises one circular chromosome and one plasmid containing several genes involved in salt adaptation and genes related to plant growth-promoting traits under salt stress. Based on previous experiments, ACC deaminase activity was identified as a main mechanism of E19 T to promote plant growth under salt stress. Interestingly, no genes classically reported to encode for ACC deaminase activity are present. In general, the E19 T genome provides information to confirm, discover, and better understand many of its previously evaluated traits involved in plant growth promotion under salt stress. Furthermore, the complete E19 T genome sequence helps to define its previously reported unclear 16S rRNA gene-based phylogenetic affiliation. Hartmannibacter forms a distinct subcluster with genera Methylobrevis , Pleomorphomonas , Oharaeibacter , and Mongoliimonas subclustered with genera belonging to Rhizobiales ., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2019 Christian Suarez et al.)

Phosphate is a macronutrient and often the limiting growing factor of many ecosystems. The aim of this work was to assess the effect of various phosphate sources on the active bacterial microbiota of barley rhizosphere and endorhiza. Barley was grown on poor soil supplemented with either Ca(H 2 PO 4 ) 2 (CaP), Gafsa rock phosphate (Gafsa), sodium hexaphytate (NaHex), or not amended (P0). RNA was extracted and cDNA synthesized via reverse transcription from both rhizosphere and endorhiza of barley roots; the obtained 16S rRNA cDNA was sequenced by Ion Torrent and analyzed with QIIME and co-occurrence network analysis. Phosphatase activity was measured in the rhizosphere. The phosphate source significantly affected alpha- and beta-diversities of the active microbiota, especially in the rhizosphere. CaP enriched the relative abundance of a broad range of taxa, while NaHex and Gafsa specifically enriched one dominant Massilia-related OTU. Co-occurrence network analysis showed that the most abundant OTUs were affected by phosphate source and, at the same time, were low connected to other OTUs (thus they were relatively "independent" from other bacteria); this indicates a successful adaptation to the specific abiotic conditions. In the rhizosphere, the phosphatase activities were correlated to several OTUs. Moreover, the phosphodiesterase/alk. phosphomonoesterase ratio was highly correlated to the dominance index of the microbiota and to the relative abundance of the dominant Massilia OTU. This study shows the differential response of the rhizosphere- and endorhiza bacterial microbiota of barley to various phosphate sources in soil, thus providing insights onto this largely unknown aspect of the soil microbiome ecology and plant-microbe interactions.

The aim of the present study was to prove the long-term efficiency of the amendments zerovalent iron grit, zeolite, and Divergan® for trace metal remediation in heavily contaminated soils and to attain a recovery of microbial functionality and diversity by remediation. For immobilization of the trace metals the amendments zerovalent iron grit, natural zeolite, and Divergan® were used. Trace metal total and mobile contents were determined and bacterial communities were assessed after a SIP experiment with 13 C-labelled wheat root by Ion-Torrent Sequencing targeting the bacterial 16S rRNA gene and two trace metal resistant genes for copper and cadmium (copA and czcA gene). The results show that the remediation effect of the three amendments is still stable after five years. The mobile trace metal contents were significantly (≤0.001) reduced in all treatments, except the Cu content in the zeolite treatment. A higher diversity in active metabolizing and growing soil bacteria was observed in remediated soils as compared to the non-remediated control, especially for the Divergan® treatment. The bacterial genera Kribbella, Glycomyces, Inquilinus, Nocardioides, and Lysobacter are the most significantly enriched genera in the 13 C fractions of the treated samples. The occurrence of bacterial families, which could be identified carrying efflux-mediated metal resistance genes for Cd/Zn and Cu, were reduced in the remediated soils as compared to the non-remediated control. The most abundant bacterial family for the copA and the czcA gene is Xanthomonadaceae. The pH-value and the trace metal concentration could be identified as key drivers of bacterial community composition, and functions in trace metal contaminated soils and remediated soils., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Recycling of process wastes will be in future an essential step to meet the demands for valuable metals of a growing market. Depending on their particle sizes incineration slags are already used to recover metals but particle size fractions below 4 mm are still difficult to recycle. Therefore, different particle size fractions (mesh size 2 and 4 mm, high energy grinded) of dry discharged slags were used for bioleaching with and without the pure cultures Acidithiobacillus ferrooxidans or Leptospirillum ferrooxidans or a mixture of Acidithiobacillus thiooxidans and Acidithiobacillus ferrooxidans in batch cultures. Regarding Al, Cr, Cu, Ni, Mn and the rare earth elements Ce, La and Er, bioleaching was significantly more successful with iron oxidizing bacteria compared to abiotic controls. Metal mobilization for Al, Cu, Mn, Cr and Er with bacteria was between 70 and 100% and for Ce, Ni and La around 50% almost after 7 days, making an industrial application for the high concentrated metals like Al and Cu feasible. In addition to the recovery of valuable metals, a reduction in cost of landfilling was identified. After treatment of the slag with the microorganisms, concentrations of harmful substances in the residues could be reduced and thus a classification in lower safety levels regarding the LAGA or EU regulations was calculated., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

A Gram-negative bacterium, strain HA7 T , was isolated from the microhabitat of common hazel (Corylus avellana L.) pollen. HA7 T was found to be an aerobic, rod-shaped, catalase-positive, oxidase-negative bacterium with an optimum growth temperature of 25 °C and pH of 7. The nearly complete 16S rRNA gene sequence of HA7 T strain showed the closest similarities to Spirosoma linguale DSM 74 T (97.4 %) and Spirosoma fluviale DSM 29961 T (97.43 %). The major fatty acids (>5 %) were C16 : 1ω5c, summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH), iso-C15 : 0 and iso-C17 : 0 3-OH. The major polar lipids were an unidentified aminophospholipid and phosphatidylethanolamine. The major respiratory quinone detected was menaquinone MK-7 (95 %). The draft genome sequence included 8 794 837 bases, which contained 3665 predicted coding sequences and had a G+C content of 47.9 mol%. The genome-based comparison between HA7 T and S. linguale DSM 74 T and S. fluviale DSM 29961 T with pairwise average nucleotide identity indicated a clear distinction, between 76.2-76.3 %. Moreover, the digital DNA-DNA relatedness of HA7 T to these strains was 26.5 and 25.1 %. Based on the differential genotypic, phenotypic and chemotaxonomic properties to closely related type strains, strain HA7 T ought to be assigned with the status of a new species, for which the name Spirosomapollinicola sp. nov. is proposed. The type strain is HA7 T (DSM 105799 T =LMG 30282 T ).

Flower pollen represents a unique microbial habitat, however the factors driving microbial assemblages and microbe-microbe interactions remain largely unexplored. Here we compared the structure and diversity of the bacterial-fungal microbiome between eight different pollen species (four wind-pollinated and four insect-pollinated) from close geographical locations, using high-throughput sequencing of the 16S the rRNA gene fragment (bacteria) and the internal transcribed spacer 2 (ITS2, fungi). Proteobacteria and Ascomycota were the most abundant bacterial and fungal phyla, respectively. Pseudomonas (bacterial) and Cladosporium (fungal) were the most abundant genera. Both bacterial and fungal microbiota were significantly influenced by plant species and pollination type, but showed a core microbiome consisting of 12 bacterial and 33 fungal genera. Co-occurrence analysis highlighted significant inter- and intra-kingdom interactions, and the interaction network was shaped by four bacterial hub taxa: Methylobacterium (two OTUs), Friedmanniella and Rosenbergiella. Rosenbergiella prevailed in insect-pollinated pollen and was negatively correlated with the other hubs, indicating habitat complementarity. Inter-kingdom co-occurrence showed a predominant effect of fungal on bacterial taxa. This study enhances our basic knowledge of pollen microbiota, and poses the basis for further inter- and intra-kingdom interaction studies in the plant reproductive organs.

A Gram-negative bacterium, designated E130 T , was isolated from rhizospheric soil of Plantago winteri Wirtg. from a natural salt meadow as part of an investigation on rhizospheric bacteria from salt-resistant plant species and evaluation of their plant growth-promoting abilities. Cells were rods, non-motile, aerobic, and oxidase and catalase positive, grew in a temperature range of between 4 and 37 °C, and in the presence of 0.5-5 % NaCl (w/v). Based on 16S rRNA gene sequence analysis, strain E130 T is affiliated within the genus Ancylobacter, sharing the highest similarity with Ancylobacter rudongensis DSM 17131 T (97.6 %), Ancylobacter defluvii CCUG 63806 T (97.5 %) and Ancylobacter dichloromethanicus DSM 21507 T (97.4 %). The DNA G+C content of strain E130 T was 65.1 mol%. Its respiratory quinones were Q-9 and Q-10 and its major polar lipids comprised phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine and unidentified phospholipid. Major fatty acids of the strains E130 T were C12 : 0, C16 : 0, C18 : 1ω7c and C19 : 0cycloω8c. The DNA-DNA relatedness of E130 T to A. rudongensis DSM 17131 T , A. defluvii CCUG 63806 T and A. dichloromethanicus DSM 21507 T was 29.2, 21.2 and 32.2 % respectively. On the basis of our polyphasic taxonomic study the new isolate represents a novel species, for which the name Ancylobacter pratisalsi sp. nov. is proposed. The type strain is E130 T (LMG 29367 T =DSM 102029 T ).

Diverse microorganisms colonise the different plant-microhabitats, such as rhizosphere and phyllosphere, and play key roles for the host. However, bacteria associated with pollen are poorly investigated, despite its ecological, commercial and medical relevance. Due to structure and nutritive composition, pollen provides a unique microhabitat. Here the bacterial abundance, community structure, diversity and colonization pattern of birch, rye, rapes and autumn crocus pollens were examined, by using cultivation, high-throughput sequencing and microscopy. Cultivated bacteria belonged to Proteobacteria, Actinobacteria and Firmicutes, with remarkable differences at species level between pollen species. High-throughput sequencing of 16S rRNA gene amplicon libraries showed Proteobacteria as the dominant phylum in all pollen species, followed by Actinobacteria, Acidobacteria and Firmicutes. Both plant species and pollination type significant influenced structure and diversity of the pollen microbiota. The insect-pollinated species possessed a more similar microbiota in comparison to the wind-pollinated ones, suggesting a levelling effect by insect vectors. Scanning electron microscopy as well as fluorescent in situ hybridisation coupled with confocal laser scanning microscopy (FISH-CLSM) indicated the tectum surface as the preferred niche of bacterial colonisation. This work is the most comprehensive study of pollen microbiology, and strongly increases our knowledge on one of the less investigated plant-microhabitats., (© 2016 Society for Applied Microbiology and John Wiley & Sons Ltd.)

From the rhizosphere of two salt tolerant plant species, Hordeum secalinum and Plantago winteri growing in a naturally salt meadow, 100 strains were isolation on enrichment media for various plant growth-promoting (PGP) functions (ACC deaminase activity, auxin synthesis, calcium phosphate mobilization and nitrogen fixation). Based on the taxonomic affiliation of the isolated bacteria and their enrichment medium 22 isolates were selected to test their growth promotion effect on the crop barley (Hordeum vulgare) under salt stress in pot experiment. In parallel the isolates were characterized in pure culture for their plant growth-promoting activities. Surprisingly the best promotors did not display a promising set of PGP activities. Isolates with multiple PGP-activities in pure culture like Microbacterium natoriense strain E38 and Pseudomonas brassicacearum strain E8 did not promote plant growth. The most effective isolate was strain E108 identified as Curtobacterium flaccumfaciens, which increased barley growth up to 300%. In pure culture strain E108 showed only two out of six plant growth promoting activities and would have been neglected. Our results highlight that screening based on pure culture assays may not be suitable for recognition of best plant growth promotion candidates and could preclude the detection of both new PGPR and new plant promotion mechanisms., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Two parallel, stable operating biogas reactors were fed with increasing amounts of maize silage to monitor microbial community changes caused by overloading. Changes of microorganisms diversity revealed by SSCP (single strand conformation polymorphism) indicating an acidification before and during the pH-value decrease. The earliest indicator was the appearance of a Methanosarcina thermophila-related species. Diversity of dominant fermenting bacteria within Bacteroidetes, Firmicutes and other Bacteria decreased upon overloading. Some species became dominant directly before and during acidification and thus could be suitable as possible indicator organisms for detection of futurity acidification. Those bacteria were related to Prolixibacter bellariivorans and Streptococcus infantarius subsp. infantarius. An early detection of community shifts will allow better feeding management for optimal biogas production., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

A phosphate-mobilizing, Gram-negative bacterium was isolated from rhizospheric soil of Plantago winteri from a natural salt meadow as part of an investigation of rhizospheric bacteria from salt-resistant plant species and evaluation of their plant-growth-promoting abilities. Cells were rods, motile, strictly aerobic, oxidase-positive and catalase-negative. Phylogenetic analysis based on 16S rRNA gene sequences showed that strain E19(T) was distinct from other taxa within the class Alphaproteobacteria. Strain E19(T) showed less than 93.5 % 16S rRNA gene sequence similarity with members of the genera Rhizobium (≤93.5 %), Labrenzia (≤93.1 %), Stappia (≤93.1 %), Aureimonas (≤93.1 %) and Mesorhizobium (≤93.0 %) and was most closely related to Rhizobium rhizoryzae (93.5 % 16S rRNA gene sequence similarity to the type strain). The sole respiratory quinone was Q-10, and the polar lipids comprised phosphatidylglycerol, phosphatidylcholine, phosphatidylethanolamine, an aminolipid and an unidentified phospholipid. Major fatty acids were C18 : 1ω7c (71.4 %), summed feature 2 (C14 : 0 3-OH and/or iso-C16 : 1; 8.3 %), C20 : 0 (7.9 %) and C16 : 0 (6.1 %). The DNA G+C content of strain E19(T) was 59.9±0.7 mol%. The capacity for nitrogen fixation was confirmed by the presence of the nifH gene and the acetylene reduction assay. On the basis of the results of our polyphasic taxonomic study, the new isolate represents a novel genus and species, for which the name Hartmannibacter diazotrophicus gen. nov., sp. nov. is proposed. The type strain of Hartmannibacter diazotrophicus is E19(T) ( = LMG 27460(T) = KACC 17263(T))., (© 2014 IUMS.)

Two motile, Gram-staining-negative, aerobic, rod-shaped bacteria designated strains E48(T) and E49(T) were isolated from the rhizosphere of Hordeum secalinum from a natural salt meadow near Münzenberg, Germany. 16S rRNA gene sequence similarity analysis revealed that strains E48(T) and E49(T) shared similarities of 97.6 % with Rheinheimera pacifica KMM 1406(T) and 98.5 % with Rheinheimera nanhaiensis E407-8(T), respectively. Major fatty acids of strain E48(T) were C16 : 0, summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) and C17 : 1ω8c, and of strain E49(T) were C16 : 0, summed feature 3 (C16 : 1ω7c and/or iso-C15 : 0 2-OH) and C18 : 1ω7c. The DNA G+C contents were 50.5 mol% (E48(T)) and 50.0 mol% (E49(T)). Strains E48(T) and E49(T) grew at 4-37 °C (optimum 28 °C) and with 0-6 % NaCl (optimum 0-3 %) and 0-5 % NaCl (optimum 0-3 %), respectively. The potential for nitrogen fixation by strains E48(T) and E49(T) was evaluated by molecular techniques and the acetylene reduction assay. The DNA-DNA hybridization, physiological and molecular data demonstrated that strains E48(T) and E49(T) represent two novel species of the genus Rheinheimera, and therefore the names Rheinheimera hassiensis sp. nov. (type strain E48(T) = LMG 27268(T) = KACC 17070(T)) and Rheinheimera muenzenbergensis sp. nov. (type strain E49(T) = LMG 27269(T) = KACC 17071(T)) are proposed.

Two Gram-reaction-negative, aerobic, nitrogen-fixing, rod-shaped bacteria, designated strains E20 and E50(T), were isolated from the rhizosphere of salt meadow plants Plantago winteri and Hordeum secalinum, respectively, near Münzenberg, Germany. Based on the 16S rRNA gene sequence analysis both strains E20 and E50(T) are affiliated with the genus Cellvibrio, sharing the highest similarity with Cellvibrio gandavensis LMG 18551(T) (96.4%) and (97.1%), respectively. Strains E20 and E50(T) were oxidase and catalase-positive, grew at a temperature range between 16 and 37 °C and in the presence of 0-5% NaCl (w/v). The DNA G+C contents were 52.1 mol% (E20) and 51.6 mol% (E50(T)). Major fatty acids of strains E20 and E50(T) were summed feature 3 (C16 : 1ω7c and/or iso-C(15 : 0) 2-OH), C(16 : 0), C(18 : 1)ω7c, C(12 : 0), C(18 : 0) and C(12 : 0) 3-OH. The DNA-DNA relatedness of the strains to Cellvibrio gandavensis LMG 18551(T) was 39% for strain E20 and 58% for strain E50(T). The nitrogen fixation capability of strains E20 and E50(T) was confirmed by the acetylene reduction assay. On the basis of our polyphasic taxonomic study, strains E20 and E50(T) represent a novel species of the genus Cellvibrio, for which the name Cellvibrio diazotrophicus is proposed. The type strain of Cellvibrio diazotrophicus is E50(T) ( = LMG 27267(T) = KACC 17069(T)). An emended description of the genus Cellvibrio is proposed based on the capability of fixing nitrogen and growth in presence of up to 5% NaCl (w/v).

Water shortage necessitated South Indian sago factory owners, extracting starch out of cassava tubers, to install biogas plants where a starch utilizing microbial community multiplies and reduces the biological oxygen demand (BOD) of the waste waters by presently about 30%. The purification efficiency of sago factory waste waters, rich in solid particles and having wide C/N ratios, around 250, through unstirred biogas plants needs to be improved. Our approach was to apply instead of animal slurry nitrate (NO3(-)) and nitrous oxide (N2O) as external N-sources anticipating a better N-distribution in the unstirred biogas plants. Estimated cell numbers, bacterial community changes, on the basis of 16S rRNA gene clone libraries and changing CO2-, CH4-, N2O releases due to the presence of nitrate or N2O suggest that acid tolerant Lactobacillus spp. dominate the biogas plant inflows (pH 3.5). They were very less or not found in the outflows (pH 7.3). Assumingly, the phyla Bacteroidetes (Prevotella spp.), Proteobacteria (Rhizobium spp., Defluvibacter sp.), Firmicutes (Megasphaera spp., Dialister spp., Clostridium spp.) and Synergistetes (Thermanaerovibrio spp.), not-detectable in the biogas plant inflows, replaced them. Anaerobes, about 400cellsml(-1) in the inflows, increased to about 10(6)cellsml(-1) in the outflows. The methane formation, as confirmed by the incubation experiments, suggests that methanogens must have been present among the anaerobes. In the biogas plant in- and outflows also about 300cellsml(-1) denitrifying bacteria and up to 10(4)cfu fungi were found. Despite the low number of denitrifying bacteria nitrate added to the biogas plant in- and outflows was widely consumed and added N2O decreased considerably. Thus, wide C/N ratios substrates like sago factory waste waters keep the N2O emissions low by using N2O either as electron acceptor or by incorporating it into the growing biomass what needs to be confirmed. The biogas plant inflow samples have emitted tentatively more CO2 and the outflow samples released more CH4., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Methanogenic communities in 200L biogas reactors containing liquid manure were investigated for 33 d. The reactors were consecutively fed with casein, starch and cream. Real-time PCR with primers targeting the gene for methyl coenzyme-M reductase (mcrA) resulted in copy numbers of up to 2.1×10(9) g dry mass(-1). Single strand conformation polymorphism (SSCP) analysis revealed a stable community consisting of few hydrogenotrophic methanogens. One of the two most abundant species was closely related to Methanospirillum hungatei, whereas the other one was only distantly related to other methanogens, with Methanopyrus kandleri being the closest cultivated relative. Most probable number (MPN) cultivations were accomplished with a sample from a 600 m(3) reactor from which all manures used in the experiments originated, and equal cell counts of ca. 10(9) g dry mass(-1) were found for cultivations with acetate, H(2) and methanol. SSCP analysis of these samples and sequencing of the DNA bands identified different hydrogenotrophic methanogens in all samples, and acetoclastic methanogens closely related to Methanosarcina mazei in the samples cultivated with acetate and methanol. As the acetoclastic species were not found in any other SSCP sample, it was supposed that the ammonia values in the manure of the laboratory biogas reactor, which ranged from 2.48 to 3.61 g NH(4)-NL(-1), inhibited the growth of the acetoclastic methanogens., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

With a growing world population and global warming, we are challenged to increase food production while reducing greenhouse gas (GHG) emissions. We studied the effects of biochar (BC) and hydrochar (HC) produced via pyrolysis or hydrothermal carbonization, respectively, on GHG fluxes in three laboratory incubation studies. In the first experiment, ryegrass was grown in sandy loam mixed with equal amounts of a nitrogen-rich peanut hull BC, compost, BC+compost, double compost, or no addition (control); wetting-drying cycles and N fertilization were applied. Biochar with or without compost significantly reduced NO emissions and did not change the CH uptake, whereas ryegrass yield was significantly increased. In the second experiment, 0% (control) or 8% (w/w) of BC (peanut hull, maize, wood chip, or charcoal) or 8% HC (beet chips or bark) was mixed into a soil and incubated at 65% water-holding capacity (WHC) for 140 d. Treatments included simulated plowing and N fertilization. All BCs reduced NO emissions by ∼60%. Hydrochars reduced NO emissions only initially but significantly increased them after N fertilization to 302% (HC-beet) and 155% (HC-bark) of the control emissions, respectively. Large HC-associated CO emissions suggested that microbial activity was stimulated and that HC was less stable than BC. In the third experiment, nutrient-rich peanut hull BC addition and incubation over 1.5 yr at high WHCs did not promote NO emissions. However, NO emissions were significantly increased with BC after NHNO addition. In conclusion, BC reduced NO emissions and improved the GHG-to-yield ratio under field-relevant conditions. However, the risk of increased NO emissions with HC addition must be carefully evaluated., (Copyright © by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.)

In the present study, bacterial communities in 200-liter biogas reactors containing liquid manure consecutively fed with casein, starch, and cream were investigated over a period of up to 33 days. A 16S rRNA gene clone library identified Bacteroidetes and Firmicutes as the most abundant bacterial groups in the starting material, at 58.9% and 30.1% of sequences, respectively. The community development of both groups was monitored by real-time PCR and single-strand conformation polymorphism (SSCP) analysis. The Firmicutes and Bacteroidetes communities were unexpectedly stable and hardly influenced by batch-feeding events. The continuous feeding of starch led to community shifts that nevertheless contributed to a stable reactor performance. A longer starving period and a change in the pH value resulted in further community shifts within the Bacteroidetes but did not influence the Firmicutes. Predominant DNA bands from SSCP gels were cloned and sequenced. Sequences related to Peptococcaceae, Cytophagales, and Petrimonas sulfuriphila were found in all samples from all experiments. Real-time PCR demonstrated the abundance of members of the phylum Bacteroidetes and also reflected changes in gene copy numbers in conjunction with a changing pH value and acetate accumulation.

Methane in the biosphere is mainly produced by prokaryotic methanogenic archaea, biomass burning, coal and oil extraction, and to a lesser extent by eukaryotic plants. Here we demonstrate that saprotrophic fungi produce methane without the involvement of methanogenic archaea. Fluorescence in situ hybridization, confocal laser-scanning microscopy and quantitative real-time PCR confirm no contribution from microbial contamination or endosymbionts. Our results suggest a common methane formation pathway in fungal cells under aerobic conditions and thus identify fungi as another source of methane in the environment. Stable carbon isotope labelling experiments reveal methionine as a precursor of methane in fungi. These findings of an aerobic fungus-derived methane formation pathway open another avenue in methane research and will further assist with current efforts in the identification of the processes involved and their ecological implications.

Twenty-two isolates of the family Enterobacteriaceae, with focus on Cronobacter isolated from infant formula and the environment of milk powder plants, were comparatively identified using API 32E (bioMérieux, Marcy l'Etoile, France), 16S rRNA gene sequencing (Accugenix, Newark, USA), and matrix-assisted laser desorption and ionization-time-of-flight mass spectrometry (MALDI-TOF MS; Mabritec, Riehen, Switzerland and AnagnosTec, Potsdam, Germany). With API 32E, 22% of the isolates were assigned to species, 64% were assigned to a genus, and 14% could not be discriminated at any taxonomic level. Both 16S rRNA gene sequencing and MALDI-TOF MS assigned 100% of the isolates to species, but the identifications based on MALDI-TOF MS results were more discriminating and unequivocal. Our data indicate that MALDI-TOF MS provides the most rapid and unambiguous identification of Cronobacter and closely related Enterobacteriaceae isolates.

Our novel approach for taxonomic identification of uncultured bacteria harboring specific physiological features in complex environmental samples combines cell collection by laser microdissection and subsequent DNA analysis. The newly developed approach was successfully tested for collection and phylogenetic characterization of polyphosphate-accumulating bacteria in activated sludge and lake sediment.

Iron(III) profiles of flooded paddy soil incubated in the greenhouse indicated oxidation of iron(II) in the upper 6 mm soil layer. Measurement of oxygen with a Clark-type microelectrode showed that oxygen was only responsible for the oxidation of iron(II) in the upper 3 mm. In the soil beneath, nitrate could be used as electron acceptor instead of oxygen for the oxidation of the iron(II). Nitrate was still available 3 mm below the soil surface, and denitrifying activity was indicated by higher concentrations of nitrite between 3 and 6 mm soil depth. Nitrate was generated by nitrification from ammonium. Ammonium concentrations increased beneath 6 mm soil depth, indicating ammonium release and diffusion from deeper soil layers. High concentrations of ammonium were also found at the surface, probably resulting from N2 fixation by cyanobacteria. Experimental adjustment of the nitrate concentration in the flooding water to 200 microM stimulated nitrate-dependent iron(II) oxidation, which was indicated by significantly lower iron(II) concentrations in soil layers in which nitrate-dependent iron(II) oxidation was proposed. Soil incubated in the dark showed high iron(III) concentrations only in the layer where oxygen was still available. In this soil, the nitrogen pool was depleted because of the lack of N2 fixation by cyanobacteria. In contrast, soil incubated in the dark with 500 microM nitrate in the flooding water showed significantly higher iron(II) and significantly lower iron(II) concentrations in the anoxic soil layers, indicating nitrate-dependent iron(II) oxidation. Anoxic incubations of soil with nitrate in the flooding water also showed high concentrations of iron(II) and low concentrations of iron(II) in the upper 3 mm. As oxygen was excluded in anoxic incubations, the high iron(III) concentrations are a sign of the activity of nitrate-dependent iron(II) oxidizers. The presence of these bacteria in non-amended soil was also indicated by the most probable number (MPN) counts of nitrate-dependent iron(II) oxidizers in the layer of 3-4 mm soil depth, which revealed 1.6 x 10(6) bacteria g(-1) dry weight.