New Delhi Metallo-β-lactamase-1(NDM-1) is an enzyme that hydrolyzes a wide range of β-lactams antibiotics, including carbapenems. The presence of the NDM-1 inhibits the potential of β–lactam antibiotics in treating infections caused by bacterial strains carrying such resistances, thus leaving minimal treatment options available. Due to this, the rapid distribution of NDM-1 harboring bacteria accounts for a significant public health menace worldwide. These bacteria have been detected in clinical specimens and environmental compartments where bacterial infections are ubiquitous. In this study, identification and absolute quantification of NDM-1 in sixteen lake sediment samples collected in and around Hyderabad, India, was carried out using a real-time quantitative polymerase chain reaction (qPCR), and the results were expressed in gene copy number/ng (nanogram) of template DNA. Thirteen samples (out of sixteen) displayed a positive signal for NDM-1 during the qPCR analysis with the highest gene copy number/ng of template DNA (71.8) being observed in the Amberpet STP. Three samples, samples from Durgamcheru lake, Kandi lake, and Singur dam, were negative for the NDM-1 during the qPCR analysis. Hierarchical clustering analysis was performed to categorize the sampling location into different clusters based on pollution sources and the observed results were expressed in the form of a dendrogram.

In recent years, Feammox has made it possible to remove NH₄⁺-N under anaerobic conditions; however, its application in practical wastewater treatment processes has not been extensively reported. In this study, an up-flow anaerobic biological filter based on limonite (Lim-UAF) was developed to facilitate long-term and stable treatment of domestic sewage. Lim-UAF achieved the highest removal efficiency of chemical oxygen demand (COD) and NH₄⁺-N at a hydraulic retention time (HRT) of 24 h (Stage II). Specifically, the COD and NH₄⁺-N content decreased from 240.8 and 30.0 mg/L to about 7.5 and 0.35 mg/L, respectively. To analyze the potential nitrogen removal mechanism, the Lim-UAF was divided into three layers according to the height of the reactor. The results showed that COD and NH₄⁺-N removal had remarkable characteristics in Lim-UAF. More than 55.0% of influent COD was removed in the lower layer (0–30 cm) of Lim-UAF, while 60.2% of NH₄⁺-N was removed in the middle layer (30–60 cm). Microbial community analysis showed that the community structure in the middle and upper layers (60–90 cm) was relatively similar, but quite different from that of the lower layer. Heterotrophic bacteria were dominant in the lower layer, whereas iron-reducing and iron-oxidizing bacteria were enriched in the upper and middle layers. The formation of secondary minerals (siderite and Fe(OH)₃) indicated that the Fe(III)/Fe(II) redox cycle occurred in Lim-UAF, which was triggered by the Feammox and NDFO processes. In summary, limonite was used to develop a single-stage wastewater treatment process for simultaneously removing organic matter and NH₄⁺-N, which has excellent application prospects in domestic sewage treatment.

Soil around the gold tailing due to the smelting process of wastewater and solid waste can lead to metal (loids) contamination, especially arsenic (As). Soil microorganisms have gradually evolved adaptive mechanisms in the process of long-term adaptation to As contamination. However, comprehensive investigations on As metabolism genes and their host microbial communities in soil profiles with different levels under long-term As contamination are lacking. There are selected three typical soil profiles (0–100 cm) with different metal (loids) contamination levels (L-low, M-moderate and H-high) around tailings in this research. It uses a Metagenomic approach to explore the adaptation mechanisms of arsenic metabolism genes and arsenic metabolism gene host microorganisms in both horizontal and vertical dimensions. The results showed that four categories of As metabolism genes were prevalent in soil profiles at different As contamination, with As reduction genes being the most abundant, followed by As oxidation genes, then respiration genes and methylation genes. The As metabolism genes arsBCR, aioE, arsPH, arrAB increased with the increase of metal (loid) contaminants concentration. Longitudinal arsA, arrA, aioA, arsM and acr3 increased in abundance in deep soil. Actinobacteria, Proteobacteria, Acidobacteria, and Chloroflexi were the dominant phylum of As metabolism gene host microorganisms. Different concentrations of metal (loid) contamination significantly affected the distribution of host As metabolism genes. Random forest prediction identified As as the most critical driver of As metabolism genes and their host microorganisms. Overall, this study provides a reference for a comprehensive investigation of the detoxification mechanisms of As metabolism microorganisms in soil profiles with different As contamination conditions, and is important for the development of As metabolism gene host microbial strains and engineering applications of microbial technologies to manage As contamination.

An increase in polycyclic aromatic hydrocarbon (PAH) pollution poses significant challenges to human and ecosystem health in the Three Gorges Reservoir (TGR) of the Yangtze River. Based on the combination of PAH analysis with qPCR and high-throughput sequencing of bacteria, 32 topsoil samples collected from 16 sites along the TGR were used to investigate the distribution and biodegradation pathways of PAHs in the water-level-fluctuation zone (WLFZ). The results indicated that the concentrations of PAHs were 43.8–228.2 and 30.8–206.3 ng/g soil (dry weight) under the high- and low-water-level (HWL and LWL) conditions, respectively. The PAH concentration in urban areas was higher than that in rural areas. Under both the HWL and LWL conditions, the abundance of the bamA gene, a biomarker of anaerobic PAH biodegradation, was significantly higher than that of the ring-hydroxylating-dioxygenase (RHD) gene, a biomarker of aerobic PAH biodegradation. The abundance of the bamA gene was significantly positively correlated with PAHs (R² = 0.8), and the biodegradation percentage of PAHs incubated anaerobically was greater than that in the aerobically incubated microcosm experiments. These data implicated a key role of the anaerobic pathway in PAH biodegradation. Co-occurrence network analysis suggested that anaerobic Anaerolineaceae, Dechloromonas, Bacteroidetes_vadin HA17 and Geobacter were key participants in the biodegradation of PAHs. The diversity analysis of functional bacteria based on the bamA gene and microcosm experiments further demonstrated that nitrate was the primary electron acceptor for PAH biodegradation. These findings provide a new perspective on the mechanism of PAH biodegradation in the TGR and knowledge that can be used to develop strategies for environmental management.

The control of agricultural pests is key to maintain economically viable crops. Increasing environmental awareness, however, is leading to more restrictive European policies regulating the use of certain pesticides due to their impact on human health and the soil system. Given this context, we evaluated the efficacy of three alternatives to the soil fumigant 1,3-dichloropropene (1,3-D), which is currently banned in Europe: two non-fumigant nematicides [oxamyl (OX) and fenamiphos (FEN)] and the soil fumigant dimethyl disulfide (DMDS). We analysed the efficiency of these pesticides against root-knot nematodes and soil fungal pathogens (determined by qPCR) as well as the soil biological quality after treatments application (estimated by enzyme activities). Among treatments, 1,3-D and DMDS significantly reduced nematode populations. FEN was more effective in sandy soil, while OX had no effect in any soil. OX and FEN had no effect on fungal pathogens, whereas DMDS reduced the abundance of Rhizoctonia solani and Fusarium solani at the root level in clay-loam soil. Soil quality decreased after treatment application but then recovered throughout the experiment, indicating the possible dissipation of the pesticides. Our findings support DMDS as a potential sustainable alternative for controlling root-knot nematodes and fungal pathogens due to its effectiveness in both studied soils, although its negative impact on soil biological quality in sandier soils must be taken into account.Main finding of the work. DMDS is a reliable alternative to 1,3-D for controlling agricultural pest but its inhibitory effect on soil enzyme activities varied according to the soil characteristics.

A molecular study on how the abundance of the dechlorinating culture KB-1 affects dechlorination rates in clay till is presented. DNA extracts showed changes in abundance of specific dechlorinators as well as their functional genes. Independently of the KB-1 added, the microbial dechlorinator abundance increased to the same level in all treatments. In the non-bioaugmented microcosms the reductive dehalogenase gene bvcA increased in abundance, but when KB-1 was added the related vcrA gene increased while bvcA genes did not increase. Modeling showed higher vinyl-chloride dechlorination rates and shorter time for complete dechlorination to ethene with higher initial concentration of KB-1 culture, while cis-dichloroethene dechlorination rates were not affected by KB-1 concentrations. This study provides high resolution abundance profiles of Dehalococcoides spp. (DHC) and functional genes, highlights the ecological behavior of KB-1 in clay till, and reinforces the importance of using multiple functional genes as biomarkers for reductive dechlorination.

Marine mammals in the Barents Sea region have among the highest levels of contaminants recorded in the Arctic and the Atlantic walrus (Odobenus rosmarus rosmarus) is one of the most contaminated species within this region. We therefore investigated the relationships bewteen blubber concentrations of lipophilic persistent organic pollutants (POPs) and plasma concentrations of perfluoroalkyl substances (PFASs) and markers of endocrine and immune functions in adult male Atlantic walruses (n = 38) from Svalbard, Norway. To do so, we assessed plasma concentrations of five forms of thyroid hormones and transcript levels of genes related to the endocrine and immune systems as endpoints; transcript levels of seven genes in blubber and 23 genes in blood cells were studied. Results indicated that plasma total thyroxine (TT4) concentrations and ratio of TT4 and reverse triiodothyronine decreased with increasing blubber concentrations of lipophilic POPs. Blood cell transcript levels of genes involved in the function of T and B cells (FC like receptors 2 and 5, cytotoxic T-lymphocyte associated protein 4 and protein tyrosine phosphatase non-receptor type 22) were increased with plasma PFAS concentrations. These results suggest that changes in thyroid and immune systems in adult male walruses are linked to current levels of contaminant exposure.

In developing countries, many urban rivers are suffering from heavy contamination by untreated sewage, which implies great microbial risks. However, information regarding the bacterial pathogen diversity and distribution in urban rivers is highly limited. In this study, 41 water samples of fifteen rivers and eight samples from two sewage treatment plants in Changzhou City of Yangtze River Delta were sampled. Next-generation sequencing and a self-built reference pathogen database were used to investigate the diversity of enteric and environmental pathogens. The results indicated that the studied urban rivers were harboring diverse potential pathogen species, which primarily included enteric pathogens in Arcobacter and Bacteroides, and environmental pathogens in Acinetobacter, Aeromonas and Pseudomonas. Quantification of twelve pathogens/indicators of interest by qPCR showed that Escherichia coli, Enterococcus faecalis, Campylobacter jejuni, Arcobacter cryaerophilus, Acinetobacter johnsonii, Acinetobacter lwoffii and Aeromonas spp. were abundant, with median values ranging from 3.30 to 5.85 log10 copies/100 mL, while Salmonella, Legionella pheumophila, Mycobacterium avium, Pseudomonas aeruginosa and Staphylococcus aureus were infrequently quantified. The pollution of nutrients and human intestinal microorganisms indicated by specific markers were found to be prevalent but with different levels in the rivers. The correlation analyses revealed that the diversity (p < 0.01) and concentrations (p < 0.05) of the enteric pathogens highly correlated to the human fecal marker abundances, which indicated that the enteric pathogens in the urban rivers were likely to have originated from domestic sewage. The environmental pathogens, which are different from the enteric ones, showed various distribution patterns, and some of them were more abundant in the rivers of rich nutrient. Our findings provide a comprehensive understanding of the bacterial pathogen distribution and influencing factors in urban rivers that are impacted by domestic sewage, thereby establishing the foundation for urban water management.

Antimicrobial resistance (AMR) is a serious problem for public and animal health, and also for the environment. Monitoring and reporting the occurrence of AMR determinants and bacteria with the potential to disseminate is a priority for health surveillance programs around the world and critical to the One Health concept. Wildlife is a reservoir of AMR, and human activities can strongly influence their resistome.The main goal of this work was to study the resistome of wild boar faecal microbiome, one of the most important game species in Europe using metagenomic and culturing approaches. The most abundant genes identified by the high-throughput qPCR array encode mobile genetic elements, including integrons, which can promote the dissemination of AMR determinants. A diverse set of genes (n = 62) conferring resistance to several classes of antibiotics (ARGs), some of them included in the WHO list of critically important antimicrobials were also detected. The most abundant ARGs confer resistance to tetracyclines and aminoglycosides. The phenotypic resistance of E. coli and Enterococcus spp. were also investigated, and together supported the metagenomic results.As the wild boar is an omnivorous animal, it can be a disseminator of AMR bacteria and ARGs to livestock, humans, and the environment. This study supports that wild boar can be a key sentinel species in ecosystems surveillance and should be included in National Action Plans to fight AMR, adopting a One Health approach.

Aquatic ecosystems and drinking water supply systems worldwide are increasingly affected by taste and odor episodes. In this study, molecular approaches including next-generation sequencing (NGS) and quantitative polymerase chain reaction (qPCR) were used to study the diversity and dynamics of cyanobacteria and 2-methylisoborneol (2-MIB)-producing cyanobacteria in Yuqiao Reservoir, a eutrophicated drinking water reservoir in Tianjin city, northern China. NGS revealed that the entire cyanobacterial community consisted of 16 genera, with Planktothrix (28.8%), Pseudanabaena (18.4%), Cylindrospermosis (7.8%), and Microcystis (7.6%) being the dominant genera, while microscopic examination identified only eight cyanobacterial genera. NGS of the 2-MIB synthesis gene revealed that Pseudanabaena and Planktothricoides were the main 2-MIB producers, with Pseudanabaena being dominant. This finding demonstrated that NGS can identify 2-MIB producers quickly and accurately and it can thus play an important role in the practical monitoring of aquatic ecology. The qPCR test showed 2-MIB synthesis gene with 4.27 × 10⁶ copies/L to 2.24 × 10⁹copies/L occurring at the three sampling sites. The mic gene copy number increased before the 2-MIB concentration increased, indicating that forecasting role in dealing with the 2-MIB concentration by gene copy number. Predicting 2-MIB by qPCR in the field must be verified with additional studies. The combination of NGS and qPCR can be an even more comprehensive method to provide early warning information to managers of reservoirs and water utilities facing taste and odor incidents. This is the first amplicon NGS dataset based on 2-MIB gene to study the diversity and dynamics of 2-MIB-producing cyanobacteria.