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Metagenomics can be used to monitor the spread of antibiotic resistance genes (ARGs). ARGs found in databases such as ResFinder and CARD primarily originate from culturable and pathogenic bacteria, while ARGs from non-culturable and non-pathogenic bacteria remain understudied. Functional metagenomics is based on phenotypic gene selection and can identify ARGs from non-culturable bacteria with a potentially low identity shared with known ARGs. In 2016, the ResFinderFG v1.0 database was created to collect ARGs from functional metagenomics studies. Here, we present the second version of the database, ResFinderFG v2.0, which is available on the Center of Genomic Epidemiology web server (https://cge.food.dtu.dk/services/ResFinderFG/). It comprises 3,913 ARGs identified by functional metagenomics from 50 carefully curated datasets. We assessed its potential to detect ARGs in comparison to other popular databases in gut, soil and water (marine + freshwater) Global Microbial Gene Catalogues (https://gmgc.embl.de). ResFinderFG v2.0 allowed for the detection of ARGs that were not detected using other databases. These included ARGs conferring resistance to beta-lactams, cycline, phenicol, glycopeptide/cycloserine and trimethoprim/sulfonamide. Thus, ResFinderFG v2.0 can be used to identify ARGs differing from those found in conventional databases and therefore improve the description of resistomes.GRAPHICAL ABSTRACTCaption: Additional use of ResFinderFG v2.0 database (composed of antibiotic resistance genes obtained with functional metagenomics) on the Center of Genomic Epidemiology webserver (https://cge.food.dtu.dk/services/ResFinderFG/), allows for more exhaustive resistome descriptions.

The recognition of microbiome inhabiting the healthy female bladder engendered the need for comprehensive characterization of the female urinary microbiome (FUM) in health and disease. Although previous studies reported FUM composition at different taxonomic levels, progress towards reliable identification at species level is highly required. The aim of this study was to comprehensively characterize bacterial species of FUM of healthy reproductive-age European women by two complementary methodologies i.e., extended culturomics and long-read third generation sequencing of near full-length 16S rRNA gene.A wide diversity of bacterial species was captured (297 species) with a median of 53 species/sample, including 16 putative uropathogens. Clustering FUM into community structure types revealed high inter-individual differences. Notably, there was not a single species common to all samples, although the Lactobacillus genus was detected in all samples. Lactobacillus crispatus, Lactobacillus iners and Lactobacillus mulieris were observed in high relative abundance in several samples as well as other species (e.g., Streptococcus agalactiae, Atopobium vaginae, Gardnerella vaginalis, Gardnerella swidsinskii), while more prevalent species were often low abundant members (e.g., Finegoldia magna). We captured remarkable richness within Corynebacterium spp. (25 species) and Lactobacillaceae (4 genera, 14 species). While amplicon sequencing allowed detection of more anaerobic species (e.g., 11 Peptoniphilus spp.), culturomics enabled the identification of recently recognized Gardnerella species and putative novel Corynebacterium species.This study provided fine-grained FUM profiling at species level and revealed detailed FUM structure, which is critical to unveil the potential relationship between specific microbiome members and urinary diseases/disorders.IMPORTANCEDespite evidence of the resident microbial community in the female lower urinary tract, bacterial species diversity and abundance in healthy women is still unclear. This study demonstrated that complementarity between optimized culture-dependent and –independent approaches is highly beneficial for comprehensive FUM species profiling by detecting higher FUM species diversity than previously reported, including identification of unreported Lactobacillaceae species and putative novel Corynebacterium species. Although some particular species were present in high relative abundance, low-abundant members were more prevalent. FUM classification into community structure types demonstrated high inter-individual differences in urinary microbiome composition among healthy women. We also report moderate correlation between culture-dependent and -independent derived data highlighting drawbacks resulting from each methodological approach. Our findings suggest that FUM bacterial diversity reported from previous studies may be underestimated. Finally, our results contribute to the fundamental knowledge of healthy FUM required for further exploration of the urinary microbiome role in urinary tract diseases.

Background To date, information on healthy female urinary microbiota is available mostly at genus level and at one time point. However, profound species-level characterization of healthy urinary microbiome and its stability over time are essential for further correct interpretation of its role in healthy urogenital tract. In this study, we investigated female urogenital microbiome (FUM) at two timepoints (within 2.5-year interval) in young asymptomatic European women. We used culturomics with accurate isolates’ identification (MALDI-TOF MS and gene markers sequencing) to understand species stability within healthy FUM. Results Extended culturomics of voided midstream urine sample pairs revealed a mean Shannon diversity index of 1.25 and mean of 19 species/sample (range 5–39 species; total of 115 species; 1830 isolates). High overall species variability between individuals was captured by beta diversity and a variety of community structure types, with the largest cluster characterized by Lactobacillus crispatus, often in combination with Gardnerella vaginalis or Gardnerella genomospecies 3. Significant FUM composition differences, related to Finegoldia magna and Streptococcus anginosus, according to smoking status were found. A high species variability within individuals (Shannon index SD > 0.5 in 7 out of 10 sample pairs) with a mean of 29% of shared species (range 9.1–41.7%) was observed. Moreover, 4 out of 10 sample pairs clustered in the same community structure type. The stable FUM sample pairs presented high abundance of Lactobacillus crispatus, Streptococcus agalactiae or Lactobacillus paragasseri and Bifidobacterium spp.. Moreover, Gardnerella vaginalis, Gardnerella genomospecies 3 or Gardnerella swidsinskii were often maintained within individuals in high abundance. Conclusions Shift in species composition at two distant timepoints was frequently observed among urogenital microbiome of European asymptomatic women. This suggests possible interchange of particular species in healthy FUM and the existence of multiple health-associated FUM compositions in certain individuals. Additionally, we provided additional evidence on resilience of particular bacterial communities and identified certain species more prone to persist in urogenital tract. This study revealed important details on the FUM composition complexity relevant for studies aiming to understand microbiota role in the urogenital tract health and for identification of eubiotic and dysbiotic FUM.

During a recent study on members of the genus

Landfarming bioremediation was performed over 2 years on soil heavily polluted with weathered oil and oil derivatives: 23200 mg kg-1 of mineral oil, 35300 mg kg-1 total hydrocarbons, and 8.65 mg kg-1 of total PAHs. During the experiment, mineral oil, total hydrocarbon and PAH concentrations decreased by approximately 53%, 27% and 72%, respectively. A GC/MS-Scan was used to identify the crude oil components that persist after bioremediation treatment of contaminated soil and the metabolites generated during this process. The data shows that in weathered-hydrocarbons contaminated soil, the number of initially detected compounds after the bioremediation process further decreased over a 2 year period, and at the same time several new compounds were observed at the end of experiment. Higher persistence was also shown for heavier n-alkanes and branched alkanes, which could be detected over a longer period of time. The analysis highlights the importance of n-alkanes, their substituted derivatives and polycyclic aromatic hydrocarbons as the most significant pollutants.

In this paper we examine the efficacy of the enhanced electrokinetic remediation of a sediment from the Tamis River contaminated with Ni and Cd. The moving anode technique and an increased cathodic compartment length were simultaneously used to enhance the technique. The removal efficacy increase appeared after the enhanced treatment for Ni (23%) and Cd (73%). The increased cathodic compartment length successfully prevented OH- ions migration in the sediment. Finally, sediment classification according to different risk assessment methodologies indicates that further remediation is required even after the enhanced treatment. Here we indicate that enhanced electroremediation resulted in the removal of efficacy increase but it is highly influenced by sediment characteristics. [Projekat Ministarstva nauke Republike Srbije, br. TR 037004 i br. III 043005]

This article presents a bench-scale investigation of nickel removal during electrokinetic (EK) remediation of sediment, and the dependency of removal efficacies upon the speciation of the metal, as influenced by the duration of the nickel-sediment interaction. The sediment used in this study was from the Grand Backa canal (Vojvodina, Republic of Serbia). The sediment used is anaerobic and the nickel pollution has been aged for several years, so it is mostly sorbed by sulphides and organic mater (57%). In EXP I, conventional EK remediation was conducted on this sediment for 7 days, but was ineffective (0% removal). EXP II investigated the influence of the duration of nickel sorption onto the sediment, by contaminating the sediment with additional nickel. In this sediment, nickel appeared mainly in the most mobile, acid-soluble fraction, and was thus available for migration towards the cathode. Consequently, conventional EK remediation of this sediment resulted in significantly better nickel removal (23%) than EXP I. During EXP III, the same spiked sediment was investigated using an increased applied current density, with no significant increase in removal efficacy. This study demonstrates that metal-sediment interaction duration affects efficacy of metal removal during EK remediation.