Reuse of sewage sludge is a general trend and land application is an essential way to reuse sludge. The outbreak of coronavirus disease has raised concerns about human pathogens and their serious threat to public health. The risk of pathogenic bacterial contamination from land application of municipal sludge has not been well assessed. The purpose of this study was to investigate the presence of pathogenic bacteria in municipal sewage sludge and to examine the survival potential of certain multidrug-resistant enteroaggregative Escherichia coli (EAEC) strain isolated from sewage sludge during heat treatment. The sewage sludge produced in the two wastewater treatment plants contained pathogenic bacteria such as pathogenic E. coli, Shigella flexneri, and Citrobacter freundii. The environmental strain of EAEC isolated from the sludge was resistant to eight types of antibiotics. It could also enter the dormant state after 4.5 h of treatment at 55 °C and regrow at 37 °C, while maintaining its antibiotic resistance. Our results indicate that the dormancy of EAEC might be why it is heat-resistant and could not be killed completely during the sludge heat treatment process. Owing to the regrowth of the dormant pathogenic bacteria, it is risky to apply the sludge to land even if the sludge is heat-treated, and there is also a risk of spreading antibiotic resistance.

Freshwater mussels are among the most endangered groups of fauna anywhere in world. The indiscriminate use of antibiotics has led to the emergence of resistant strains. These antibiotic-resistant bacteria play a key role in increasing the risk allied with the use of surface water and in spread of resistance genes. Two endangered freshwater mussel species, Margaritifera margaritifera and Potomida littoralis, were sampled at 4 sampling sites along a 50 km stretch of River Tua. Water samples were taken at same sites. Of the total of 135 isolates, 64.44% (39.26% from water and 25.19% from mussels) were coliform bacteria. Site T1, with the lowest concentration of coliform bacteria, and site T2 were the only ones where M. margaritifera was found. No E. coli isolates were found in this species and the pattern between water and mussels was similar. P. littoralis, which was present at T3/T4 sites, is the one that faces the highest concentration of bacterial toxins, which are found in treated wastewater effluents and around population centers. Sites T3/T4 have the isolates (water and mussels) with the highest resistance pattern, mainly to β-lactams. Water and P. littoralis isolates (T3/T4) showed resistance to penicillins and their combination with clavulanic acid, and to cephalosporins, precisely to a fourth generation of cephalosporin antibiotics. The analysis provides important information on the risk to water systems, as well as the need to investigate possible management measures. It is suggested that future studies on the health status of freshwater bivalves should incorporate measures to indicate bacteriological water quality.

The intestinal microbiota has a key role in human health via the interaction with the somatic and immune cells in the digestive tract environment. Food, through matrix effect, nutrient and non-nutrient molecules, is a key regulator of microbiota diversity. As a food contaminant, the pesticide chlorpyrifos (CPF) has an effect on the composition of the intestinal microbiota and induces perturbation of microbiota. Prebiotics (and notably inulin) are known for their ability to promote an equilibrium of the microbiota that favours saccharolytic bacteria. The SHIME® dynamic in vitro model of the human intestine was exposed to CPF and inulin concomitantly for 30 days, in order to assess variations in both the bacterial populations and their metabolites. Various analyses of the microbiota (notably temporal temperature gradient gel electrophoresis) revealed a protective effect of the prebiotic through inhibition of the enterobacterial (E. coli) population. Bifidobacteria were only temporarily inhibited at D15 and recovered at D30. Although other potentially beneficial populations (lactobacilli) were not greatly modified, their activity and that of the saccharolytic bacteria in general were highlighted by an increase in levels of short-chain fatty acids and more specifically butyrate. Given the known role of host-microbiota communication, CPF's impact on the body's homeostasis remains to be determined.

Bacteria are candidates for the biotransformation of environmental arsenic (As), while As metabolism in bacteria is not yet fully understood. In this study, we sequenced the genome of an As-resistant bacterium strain Stenotrophomonas maltophilia SCSIOOM isolated from the fish gut. After arsenate (As(V)) exposure, S. maltophilia transformed As(V) to organoarsenicals, along with the significant change of the expression of 40 genes, including the upregulation of arsH, arsRBC and betIBA. The heterogeneous expression of arsH and arsRBC increased As resistance of E. coli AW3110 by increasing As efflux and transformation. E. coli AW3110 (pET-betIBA) could transform inorganic As into dimethylarsinate (DMA) and nontoxic arsenobetaine (AsB), which suggested that AsB could be synthesized through the synthetic pathway of its analog-glycine betaine. In addition, the existence of arsRBC, betIBA and arsH reduced the reactive oxygen species (ROS) induced by As exposure. In total, these results demonstrated that S. maltophilia adopted an As metabolism strategy by reducing As accumulation and synthesizing less toxic As species. We first reported the production and potential synthetic pathway of AsB in bacteria, which improved our knowledge of As toxicology in microorganisms.

The bacteria toxicity of nanoscale zero-valent iron (nZVI) can be changed during its application in water treatment but the toxicity mechanism is still not well understood, particularly under anaerobic conditions. Here, the toxicity of nZVI and its aging products towards Escherichia coli (E. coli) and the mechanisms of extracellular and intracellular reactive oxygen species (ROS) damage were deeply probed in the presence and absence of oxygen in ultrapure water. Under aerobic conditions, the ROS damage primarily caused by the generation of extracellular free •OH can be a major contributor to the toxicity of nZVI to E. coli. By contrast, in anaerobic nZVI treatment system, the intracellular •OH can be quenched by benzoic acid which is a cell permeable quencher and the electron spin resonance (ESR) signals of 5,5-dimethy-1-pyrroline (DMPO)- •OH were evidently observed in system with the addition of F⁻ which could desorb the surface •OH into solution. It indicated that the intracellular •OH adsorbed on the particle surface can also play an indispensable role in inactivating cells under anaerobic conditions. Moreover, nZVI can steeply decline the membrane potential, causing severe membrane disruption and therefore resulting in the stronger toxicity in anaerobic conditions. Furthermore, the chemical composition transformation of nZVI and generation of benign iron corrosion products (e.g., Fe₃O₄, γ-Fe₂O₃, γ-FeOOH) are mainly responsible for the reduced toxicity with the increasing aging time. These results provide insights into the extracellular and intracellular ROS damage occurred in aerobic and anaerobic nZVI treatment systems, offering more perspective to the risk assessment of nZVI application.

The black soldier fly (BSF) Hermetia illucens has a strong tolerance to cadmium stress. This helps to use BSF in entomoremediation of heavy metal pollution. Rich metallothionein (MT) proteins were thought to be important for some insects to endure the toxicity of heavy metal. We identified and characterized three MTs genes in BSF (BSFMTs), including BSFMT1, BSFMT2A, and BSFMT2B. Molecular modeling was used to predict metal binding sites. Phylogenetic analysis was used to identify gene families. Overexpression of the recombinant black soldier fly metallothioneins was found to confer Cd tolerance in Escherichia coli. Finally, functions of BSFMTs in BSF were explored through RNA interference (RNAi). RNAi results of BSFMT2B showed that the larval fresh weight decreased significantly, and the larvae mortality increased significantly. This study suggests that BSFMTs have important properties in Cd detoxification and tolerance in BSF. Further characterization analyses of physiological function about metallothioneins are necessary in BSF and other insects.

The treatment of industrial waste and harmful bacteria is an important topic due to the release of toxins from the industrial pollutants that damage the water resources. These harmful sources frighten the life of every organism which was later developed as the carcinogenic and mutagenic agents. Therefore, the current study focuses on the breakdown or degradation of 4-chlorophenol and the antibacterial activity against Escherichia coli (E. coli). As a well-known catalyst, pure titanium-di-oxide (TiO₂) had not shown the photocatalytic activity in the visible light region. Hence, band position of TiO₂ need to be shifted to bring out the absorption in the visible light region. For this purpose, the n-type TiO₂ nanocrystalline material's band gap got varied by adding different ratios of p-type CuO. The result had appeared in the formation of p (CuO) – n (TiO₂) junction synthesized from sol-gel followed by chemical precipitation methods. The optical band gap value was determined by Kubelka-Munk (K-M) plot through UV–Vis diffusive reflectance spectroscopy (DRS). Further, the comprehensive mechanism and the results of photocatalytic and antibacterial activities were discussed in detail. These investigations are made for tuning the TiO₂ catalyst towards improving or eliminating the existing various environmental damages.

The environment is considered a major reservoir of antimicrobial resistant microorganisms (AMR) and antimicrobial resistance genes (ARG). Colistin, a “last resort” antibiotic, is used for the treatment of severe infections caused by multidrug-resistant Gram-negative bacteria. The global dissemination of mobile colistin resistance genes (mcr) in natural and non-natural environments is a major setback in the fight against antimicrobial resistance. Hitherto, there is a limited number of studies screening this resistance determinant in bacteria from wildlife. In this study, we describe for the first time the detection of plasmid-mediated colistin resistance in Escherichia coli from wild ungulates in Portugal, which are also widely distributed across Europe. This information is critical to identify the importance of ungulates in the dissemination of resistant bacteria, and their corresponding genes, across the environment. Here, 151 resistant-Enterobacteriaceae isolated from 181 samples collected from different wild ungulate species throughout Portugal were screened for mcr genes. Four mcr-1-positive Escherichia coli were detected from four fallow deer individuals that were sampled in the same hunting ground. These four isolates harboured mcr-1-related IncP plasmids belonging to sequencing types ST155, ST533 and ST345 (n = 2), suggesting bacterial and/or plasmid circulation. All mcr-1-positive E. coli also showed other resistance phenotypes, including MDR, including the B1 commensal phylogenetic profile. All mcr-1-positive E. coli show additional resistance phenotypes, including MDR, including the B1 commensal phylogenetic profile. Our findings are upsetting, highlighting the global dissemination of colistin resistance genes in the whole ecosystem, which, under the One Health framework, emphasizes the urgent need for effective implementation of AMR surveillance and control in the human-animal-environment interfaces.

Microplastics (MPs; <5 mm) released into freshwaters from anthropogenic sources accumulate in sediments, where they may pose an environmental threat to benthic organisms, such as nematodes. Several studies have examined the effects of nano- and microplastics on the nematode Caenorhabditis elegans, whereas reduced food availability was suggested as a possible explanation for the observed inhibitory effects. Therefore, this study should clarify whether micro-beads of different sizes (1.0 and 6.0 μm in diameter) and materials (polystyrene PS, silica) are able to interfere with the feeding of C. elegans on its bacterial diet (Escherichia coli), and, by this, lowering its consumption rate within 7 h of exposure. Moreover, it was examined whether an inhibited bacterial consumption was caused by a reduction of the nematode’s pumping rate, as a primary indicator of food ingestion. Bacterial consumption by C. elegans was significantly decreased in the presence of 1.0- and 6.0-μm PS beads (49–67% lower bacterial consumption compared to control), whereas in the presence of 1.0-μm silica beads feeding was not impeded. Interestingly, the pumping rate was significantly lower in the presence of non-ingestible 6.0-μm PS beads with 161 ± 16 pumps min⁻¹, while it was largely unchanged for nematodes exposed to ingestible 1.0-μm PS beads with 205 ± 12 pumps min⁻¹, compared to control conditions with 210 ± 18 pumps min⁻¹, respectively. As reduced bacterial consumption leads to generally lower energy reserves in C. elegans, these results allow to link observed inhibitory effects of MPs on the nematodes to a lower food availability. Such indirect, food-web related, effects of MPs should raise concern of ecological consequences in natural habitats, where temporal food deficiencies can occur. Consequently, disturbances in food availability and feeding efficiency should be regarded as important parameters in environmental risk assessments focusing on MPs.

The spread of antibiotic resistance genes (ARGs) is a global environmental issue. Dissolved biochar is more likely to contact bacteria in water, producing ecological risks. This study explored the effects of dissolved biochar on ARGs transfer in bacteria. Conjugative transfer efficiency was significantly different following treatment with different types of dissolved biochar. Typically, humic acid-like substance in dissolved biochar can significantly improve the transfer efficiency of ARGs between bacteria. When the concentration of dissolved biochar was ≤10 mg biochar/mL, humic acid-like substance substantially promoted ARGs transfer. An increase in dissolved biochar concentration weakened the ARGs transfer from humic acid-like substance. The inhibitory effects of small-molecule matters dominated, decreasing conjugative transfer frequency. At a concentration of 100 mg biochar/mL, the conjugative transfer efficiency of all treatments was lower than that of control. Compared with corn straw dissolved biochar, there were more transconjugants in pine sawdust dissolved biochar. Following treatment with 10 mg biochar/mL pine sawdust dissolved biochar, the number of transconjugants was at its maximum; approximately 7.3 folds higher than the control. We also explored mechanisms by which dissolved biochar impacts conjugative transfer. Due to the complex composition of dissolved biochar, its effects on the expression of conjugative transfer-related genes were also dynamic. This study investigates the ecological risk of biochar and guides its scientific application.