Long-term and combined heavy-metal contamination forms a unique microbiome and resistome: A case study in a Yellow River tributary sediments.

Microorganisms have developed mechanisms to adapt to environmental stress, but how microbial communities adapt to long-term and combined heavy-metal contamination under natural environmental conditions remains unclear. Specifically, this study analyzed the characteristics of heavy metal composition, microbial community, and heavy metal resistance genes (MRGs) in sediments along Mang River, a tributary of the Yellow River, which has been heavily polluted by industrial production for more than 40 years. The results showed that the concentrations of Cr, Zn, Pb, Cu and As in most sediments were higher than the ambient background values. Bringing the heavy metals speciation and concentration into the risk evaluation method, two-thirds of the sediment samples were at or above the moderate risk level, and the ecological risk of combined heavy metals in the sediments decreased along the river stream. The high ecological risk of heavy metals affected the microbial community structure, metabolic pathways and MRG distribution. The formation of a HM-resistant microbiome possibly occurred through the spread of insertion sequences (ISs) carrying multiple MRGs, the types of ISs carrying MRGs outnumber those of plasmids, and the quantity of MRGs on ISs is also higher than that on plasmids. These findings could improve our understanding of the adaptation mechanism of microbial communities to long-term combined heavy metal contamination.

Development of a turn-on fluorescent probe for the imaging of intracellular hypochlorous acid (HClO) during ferroptosis.

Ferroptosis is a burgeoning iron-dependent cell death form, and has close relation with hypochlorous acid (HClO). Exploring the fluctuation of the HClO level in living cells during ferroptosis could contribute to the profound study of the biological functions of HClO during ferroptosis. Here, we present a turn-on probe (RH-C) for the imaging of intracellular HClO during ferroptosis. The probe RH-C utilized the N,N-dimethylthiocarbamate group as a selective recognition site for HClO, and displayed desirable sensitivity and selectivity to HClO. The probe RH-C could detect the exogenous and endogenous HClO in living cells. Furthermore, RH-C was competent in monitoring the changes of endogenous HClO level during the process of ferroptosis. Biological imaging results suggested that erastin-induced ferroptosis can result in the excessive production of the endogenous HClO, and ferrostatin-1 (Fer-1) and vitamin E (VE) could block the massive accumulation of HClO in living cells.

PLEKHM2 deficiency induces impaired mitochondrial clearance and elevated ROS levels in human iPSC-derived cardiomyocytes.

Pleckstrin homology domain-containing family M member 2 (PLEKHM2) is an essential adaptor for lysosomal trafficking and its homozygous truncation have been reported to cause early onset dilated cardiomyopathy (DCM). However, the molecular mechanism of PLEKHM2 deficiency in DCM pathogenesis and progression is poorly understood. Here, we generated an in vitro model of PLEKHM2 knockout (KO) induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to elucidate the potential pathogenic mechanism of PLEKHM2-deficient cardiomyopathy. PLEKHM2-KO hiPSC-CMs developed disease phenotypes with reduced contractility and impaired calcium handling. Subsequent RNA sequencing (RNA-seq) analysis revealed altered expression of genes involved in mitochondrial function, autophagy and apoptosis in PLEKHM2-KO hiPSC-CMs. Further molecular experiments confirmed PLEKHM2 deficiency impaired autophagy and resulted in accumulation of damaged mitochondria, which triggered increased reactive oxygen species (ROS) levels and decreased mitochondrial membrane potential (Δψm). Importantly, the elevated ROS levels caused oxidative stress-induced damage to nearby healthy mitochondria, resulting in extensive Δψm destabilization, and ultimately leading to impaired mitochondrial function and myocardial contractility. Moreover, ROS inhibition attenuated oxidative stress-induced mitochondrial damage, thereby partially rescued PLEKHM2 deficiency-induced disease phenotypes. Remarkably, PLEKHM2-WT overexpression restored autophagic flux and rescued mitochondrial function and myocardial contractility in PLEKHM2-KO hiPSC-CMs. Taken together, these results suggested that impaired mitochondrial clearance and increased ROS levels play important roles in PLEKHM2-deficient cardiomyopathy, and PLEKHM2-WT overexpression can improve mitochondrial function and rescue PLEKHM2-deficient cardiomyopathy.

Green synthesis of a chlorfenapyr chitosan nanopesticide for maize root application: Reducing environmental pollution and risks to nontarget organisms.

Chlorfenapyr (CHL) is a pyrrole insecticide with a novel structure that is used to control resistant pests. However, its weak systemic activity limits its application to crop roots. Herein, a novel CHL formulation with improved effective utilization rates and suitability for root application is developed to avoid or reduce contamination caused by pesticide spraying. Accordingly, we prepared CHL@CS/CMCS nanoparticle (NP) suspensions with a particle size of approximately 100 nm using chitosan (CS) and carboxymethyl chitosan (CMCS). These suspensions exhibited better thermal stability, adhesion, permeability and systemic activity than a CHL suspension concentrate (CHL-SC). The nanoformulation deposition rate on maize leaves after spraying was 12.28 mg/kg, significantly higher than that of CHL-SC. The nanosuspension was effectively absorbed and transported by roots after irrigation and was suitable for root application. The efficacy was 89.46-92.36 % against Spodoptera frugiperda at 7 d, 7.5-17.5 times higher than that of CHL-SC. Furthermore, the CHL@CS/CMCS nanosuspension was safer for earthworms. These results suggest that chitosan-based nanoformulations improve the efficacy, utilization efficiency and active period of CHL control, providing a new approach for CHL application, reducing pollutant dispersal and the environmental impacts of pesticide application and facilitating sustainable agricultural production.

Improved Calculations of Heavy Metal Toxicity Coefficients for Evaluating Potential Ecological Risk in Sediments Based on Seven Major Chinese Water Systems.

Several methods have been used to assess heavy metal contamination in sediments. However, an assessment that considers both composite heavy metal speciation and concentration is necessary to accurately study ecological risks. This study improved the potential ecological risk index method and calculated the toxicity coefficients of seven heavy metals: Arsenic (As), Cadmium (Cd), Chromium (Cr), Copper (Cu), Nickel (Ni), Lead (Pb), and Zinc (Zn). The newly calculated toxicity coefficients were validated by using previously published heavy metal distribution data of the Henan section of the Yellow River. The calculation procedure is based on the principle that the abundance of heavy metals in the environment and their bioavailable forms affect the toxicity of heavy metals. The toxicity coefficients for the seven heavy metals were calculated as follows: As = 10, Cd = 20, Cr = 5, Cu = 2, Ni = 5, Pb = 5, Zn = 1. Ecological risk assessment of the Henan section of the Yellow River using the improved toxicity coefficients revealed that the ecological risk of Cd and total heavy metals is higher than previous calculations, reaching the strength and moderate risk levels, respectively. The improved potential ecological risk index method is more sensitive to heavy metal pollution and thus provides a better indication of ecological risk. This is a necessary improvement to provide more accurate pollution assessments.

Nano-pesticide carrier O-Carboxymethyl chitosan is indigestible in Apis cerana cerana and affects intestinal flora.

O-Carboxymethyl chitosan nanoparticles (O-CMC-NPs), which are organic pesticide carriers, have excellent application potential. Exploring the effects of O-CMC-NPs on non-target organisms, such as Apis cerana cerana, is critical for their effective application; however, such studies are limited. This study investigated the stress response of A. cerana Fabricius after O-CMC-NPs ingestion. The administration of high O-CMC-NP concentrations enhanced the activities of antioxidant and detoxifying enzymes in A. cerana, with the activity of glutathione-S-transferase increasing by 54.43 %-64.33 % after one day. The transit of O-CMC-NPs into the A. cerana midgut resulted in their deposition and adherence to the intestinal wall, as they cluster and precipitate in acidic conditions. The population of Gillianella bacteria in the middle intestine was remarkably reduced after 6 d of administration of high O-CMC-NP concentrations. Contrastingly, the abundance of Bifidobacteria and Lactobacillus in the rectum significantly increased. These results indicate that the intake of high concentrations of O-CMC-NPs causes a stress response in A. cerana and affects the relative abundance of crucial intestinal flora, which may pose a potential risk to the colony. This implies that even nanomaterials with favorable biocompatibility should be applied reasonably within a specific range to avoid adverse effects on the environment and non-target organisms in the context of large-scale research and promotion of nanomaterials.

Rotenone encapsulated in pH-responsive alginate-based microspheres reduces toxicity to zebrafish.

Rotenone is a botanical pesticide and has long been used for control of insect pests and also as a natural piscicide for management of fish populations in many countries. Field application for pest control, however, often encounters the movement of rotenone into surface water due to spray drift or surface runoff after rainfall, which could potentially result in water pollution and unexpected death of fishes. To minimize its effect on freshwater and the problem of fish dying, one solution was to encapsulate rotenone in specific microspheres, limiting its release and reducing its toxicity since rotenone can be quickly degraded under sunlight. In this study, pH-responsive alginate-based microspheres were synthesized to encapsulating rotenone, which were designated as rotenone beads. The rotenone beads, along with alginate beads (devoid of rotenone) were characterized and evaluated for their responses to pH and effects on zebrafish. Results showed that the microspheres had high loading efficiency (4.41%, w/w) for rotenone, and rotenone beads well responded to solution pH levels. The cumulative release rates of rotenone from the beads were 27.91%, 42.72%, and 90.24% at pH 5.5, 7.0, and 9.0, respectively. Under acidic conditions, the rotenone release rate was lower due to hydrogen bonding. On the contrary, rotenone became more quickly released at the high pH due to intermolecular repulsion. The toxicity of rotenone beads to zebrafish and fish embryos at a pH of 5.5 was reduced by 2- and 4-fold than chemical rotenone. Since pH levels in most freshwater lakes, ponds, and streams vary from 6 to 8, rotenone release from the beads in such freshwater could be limited. Thus, the synthesized rotenone beads could be relatively safely used for pest control with limited effects on freshwater fishers.

Novel application of biodegradable chitosan in agriculture: Using green nanopesticides to control Solenopsis invicta.

Botanical pesticides are biological pesticides that are environment friendly. However, their instability and short persistence limit their application. In this study, pH sensitive chitosan based rotenone (Rot) nanoparticles (CS/CMCS/Rot-NPs) were prepared using chitosan and carboxymethyl chitosan to take advantage of the acidic nature of the red fire ant midgut. Chitosan based nanoparticles showed photoprotective and slow sustained release effects on Rot and significantly increased the insecticidal activity of Rot against red fire ants. The 24-96hLC50 of CS/CMCS/Rot-NPs against red fire ants was 3.28-6.84 fold that of Rot. The CS/CMCS/Rot-NPs significantly reduced the venom alkaloid content of red fire ants and their living environment and weakened their survival by increasing their survival cost in the ecological environment. Nanotechnology combined with botanical pesticides can be used as a novel, safe, effective, and ecofriendly method to control red fire ants.

Becoming a Doctor During a Pandemic: Impact on Medical Student Social Identity Formation.

The COVID-19 pandemic required modifications to undergraduate medical education that likely affected medical students' social identity formation (SIF). SIF is shaped by experiences throughout the medical education continuum. This commentary explores factors potentially affecting medical student SIF during the COVID-19 pandemic focusing on students' perceptions of being part of the healthcare team, their role in medicine, and their engagement during the pandemic. Based on such considerations, we propose that educators should aim to design effective learning environments to support a full educational experience that encompasses acquiring medical knowledge and building strong social identities even during a pandemic.

Industrial aerobic composting and the addition of microbial agents largely reduce the risks of heavy metal and ARG transfer through livestock manure.

Composting is an effective and necessary modality in the recycling of agricultural wastes such as livestock manure, furfural, and straw. However, the risks of heavy metals (HMs) and antibiotic resistance genes (ARGs) during industrial-scale composting process have not been adequately assessed, especially with the addition of bacterial agents. In this study, changes in HMs toxicity, ARGs propagation and microbial community structure during industrial-scale aerobic composting of livestock manure were firstly investigated with various substrates addition. Moreover, the effect of the addition of bacterial agents (Bacillus strains) was evaluated. The results showed that industrial aerobic composting process could immobilize various HMs with different extents and significantly reduce the levels of ARGs such as intl1 and oqxB genes. The addition of Bacillus strains could further reduce the levels of most detected ARGs and the bioavailability of Cu and Cr, and the relative abundance of ereA and tetA was undetectable in some materials. After composting, the main bacterial community structures were similar among different substrates irrespective of bacterial agents and indicated that the composting process was the main driver for their change. This study provides a scientific reference for the safe reuse of livestock manure.

The combination of thymol and cinnamaldehyde reduces the survival and virulence of Listeria monocytogenes on autoclaved chicken breast.

AIMS: To reveal the antibacterial mechanism of the combination of thymol and cinnamaldehyde to Listeria monocytogenes ATCC 19115 on autoclaved chicken breast. METHODS AND RESULTS: In this study, L. monocytogenes ATCC 19115 on autoclaved chicken breast was exposed to the stress of 125 µg/ml thymol and 125 µg/ml cinnamaldehyde, and transcriptome analysis was used to reveal the crucial antibacterial mechanism. According to the results, 1303 significantly differentially expressed genes (DEGs) were identified. Treated by thymol and cinnamaldehyde in combination, pyrimidine and branched-chain amino acid biosynthesis of L. monocytogenes were thwarted which impairs its nucleic acid biosynthesis and intracellular metabolism. The up-regulated DEGs involved in membrane composition and function contributed to membrane repair. Besides, pyruvate catabolism and TCA cycle were restrained which brought about the disturbance of amino acid metabolism. ABC transporters were also perturbed, for instance, the uptake of cysteine, D-methionine, and betaine was activated, while the uptake of vitamin, iron, and carnitine was repressed. Thus, L. monocytogenes tended to activate PTS, glycolysis, glycerol catabolism, and pentose phosphate pathways to obtain energy to adapt to the hostile condition. Noticeably, DEGs involved in virulence factors were totally down-regulated, including genes devoted to encoding flagella, chemotaxis, biofilm formation, internalin as well as virulence gene clusters. CONCLUSIONS: The combination of thymol and cinnamaldehyde is effective to reduce the survival and potential virulence of L. monocytogenes on autoclaved chicken breast. SIGNIFICANCE AND IMPACT OF STUDY: This work contributes to providing theoretical information for the application and optimization of thymol and cinnamaldehyde in ready-to-eat meat products to inhibit L. monocytogenes.

A review on slip boundary conditions at the nanoscale: recent development and applications.

The slip boundary condition for nanoflows is a key component of nanohydrodynamics theory, and can play a significant role in the design and fabrication of nanofluidic devices. In this review, focused on the slip boundary conditions for nanoconfined liquid flows, we firstly summarize some basic concepts about slip length including its definition and categories. Then, the effects of different interfacial properties on slip length are analyzed. On strong hydrophilic surfaces, a negative slip length exists and varies with the external driving force. In addition, depending on whether there is a true slip length, the amplitude of surface roughness has different influences on the effective slip length. The composition of surface textures, including isotropic and anisotropic textures, can also affect the effective slip length. Finally, potential applications of nanofluidics with a tunable slip length are discussed and future directions related to slip boundary conditions for nanoscale flow systems are addressed.

Enhanced Control of the Fungus Gnat Bradysia odoriphaga (Diptera: Sciaridae) by Co-Application of Clothianidin and Hexaflumuron.

The fungus gnat is a major pest of chive in China. Its control has been relied heavily on the application of clothianidin. Due to the intensive application, its control efficacy become reduced. The present study was intended to evaluate co-drenching of clothianidin with hexaflumuron on absorption and dissipation of clothianidin in chive plants and soils and determine the effect of such application on control efficacies. Chive production fields in Guangdong and Hubei Provinces were drenched with clothianidin alone and a mixture of clothianidin and hexaflumuron at low application rates. Concentrations of clothianidin in chive plants and soils were analyzed by HPLC. Results showed that co-application had higher control efficacies against the fungus gnat than clothianidin alone. The co-application enhanced clothianidin absorption and dissipation and extended the half-lives of clothianidin in chive. It was likely that hexaflumuron protected chive roots from larva damage, and healthy roots absorbed more clothianidin, resulting in the extension of the half-lives. Additionally, the terminal residues of clothianidin in chive after 14 days of application were lower than the maximum residue limit in chive set by the Codex Alimentarius Commission. This study for the first time documented that co-application of clothianidin and hexaflumuron improved chive plants in absorption and dissipation of clothianidin and enhanced fungus gnat control efficacies.

Floating chitosan-alginate microspheres loaded with chlorantraniliprole effectively control Chilo suppressalis (Walker) and Sesamia inferens (Walker) in rice fields.

Striped rice stem borer, Chilo suppressalis (Walker) and pink stem borer, Sesamia inferens (Walker) are two important pests, causing substantial yield loss in rice production. Application of conventional synthetic pesticides, such as suspension concentrates and water-dispersible granules, is a primary method for control of the two pests. Due to the flow of water in rice field, spray drift, and soil adsorption, applied such pesticides are often out of the target, resulting in low control efficacy, potential contamination of soil or surface water, and also threat to human health. Thus, there is an urgent need for developing environmentally friendly and highly targeted pesticide formulations to meet the challenges. The present study synthesized chlorantraniliprole loaded chitosan-alginate floating hydrogel microspheres (CCAM) through physical embedding, ionic crosslinking, and incorporation of citronellol as an oil phase. The morphology, particle size, entrapment efficiency, loading capacity, in vitro slow-release kinetics, and floating ability of the CCAM were tested in laboratory conditions. The CCAM and two commercial formulations (suspended and granulated) of chlorantraniliprole were respectively evaluated in two rice fields located in two provinces of China. The CCAM was able to float on the surface of rice field, gather around rice stems, and slowly release chlorantraniliprole, which resulted in significantly higher concentrations of chlorantraniliprole in rice stems and leaves for a prolonged time than suspended and granulated controls. The application of CCAM provided an on-target control of both striped stem borer and pink stem borer. Furthermore, CCAM application had very low residue of chlorantraniliprole in soils. As far as is known, this is the first report of chlorantraniliprole loaded on chitosan-alginate floating hydrogel microspheres for rice stem borer control. Our results indicate that the synthesized CCAM could potentially be used as a controlled-release product for effective control of the two rice pests, while reducing the residual chlorantraniliprole in the soil and avoiding pesticide drift.

Analysis of methods for quantifying yeast cell concentration in complex lignocellulosic fermentation processes.

Cell mass and viability are tightly linked to the productivity of fermentation processes. In 2nd generation lignocellulose-based media quantitative measurement of cell concentration is challenging because of particles, auto-fluorescence, and intrinsic colour and turbidity of the media. We systematically evaluated several methods for quantifying total and viable yeast cell concentrations to validate their use in lignocellulosic media. Several automated cell counting systems and stain-based viability tests had very limited applicability in such samples. In contrast, manual cell enumeration in a hemocytometer, plating and enumeration of colony forming units, qPCR, and in situ dielectric spectroscopy were further investigated. Parameter optimization to measurements in synthetic lignocellulosic media, which mimicked typical lignocellulosic fermentation conditions, resulted in statistically significant calibration models with good predictive capacity for these four methods. Manual enumeration of cells in a hemocytometer and of CFU were further validated for quantitative assessment of cell numbers in simultaneous saccharification and fermentation experiments on steam-exploded wheat straw. Furthermore, quantitative correlations could be established between these variables and in situ permittivity. In contrast, qPCR quantification suffered from inconsistent DNA extraction from the lignocellulosic slurries. Development of reliable and validated cell quantification methods and understanding their strengths and limitations in lignocellulosic contexts, will enable further development, optimization, and control of lignocellulose-based fermentation processes.

Residue and dissipation of two formulations of emamectin benzoate in tender cowpea and old cowpea and a risk assessment of dietary intake.

The dissipation and residue levels of emamectin benzoate emulsifiable concentrate (EC) and microemulsion (ME) formulations in tender cowpeas and old cowpeas were investigated under field conditions. The decline curves of emamectin benzoate residues in cowpea corresponded to first-order kinetics. The dissipation rate of emamectin benzoate in tender cowpeas was faster than that in old cowpeas. The half-lives of the EC were 1.34-1.39 d and 1.74-2.31 d in tender cowpea and old cowpea, respectively. For the ME, the half-lives were 1.39-1.51 d and 2.08-2.67 d, respectively. The risk of adult intake of emamectin benzoate from cowpea is within the acceptable limits of the human body. Compared to tender cowpeas, the risk of eating old cowpeas is higher. Emamectin benzoate (EC) is recommended for cowpeas when the intention is to harvest tender cowpeas, while both formulations are acceptable for cowpeas when the intention is to harvest old cowpeas.

Insecticidal efficacy and mechanism of nanoparticles synthesized from chitosan and carboxymethyl chitosan against Solenopsis invicta (Hymenoptera: Formicidae).

The efficacy and mode of action of biodegradable chitosan (CS) and carboxymethyl chitosan (CMCS) organic polymer nanoparticles (NPs) on insects were studied. The prepared CS/CMCS-NPs were spherical with a particle size of 142.1 ± 2.0 nm. The swelling test showed that they were pH-sensitive, and the swelling rate was 554 % at pH 4.5. It was found that CS/CMCS-NPs had insecticidal efficacy against red fire ants (S. invicta). The mortality of red fire ants on the 6th day after treatment with 0.2 % and 0.06 % CS/CMCS-NPs suspensions was 98.33 ± 1.67 % and 48.33 ± 3.33 %, respectively. After CS/CMCS-NPs treatment, the food intake, growth, and development of red fire ants were inhibited; the midgut was significantly expanded; and the activity of digestive enzymes in the midgut was decreased. Our findings suggest that CS/CMCS-NPs mainly inhibited the digestion function of the midgut, leading to the death of red fire ants.

Responses of bacterial and bacteriophage communities to long-term exposure to antimicrobial agents in wastewater treatment systems.

The occurrence of antibacterial agents has received increasing concern due to their possible threats to human health. However, the effects of antibacterial residues on the evolution and dynamics between bacteria and bacteriophages in wastewater treatment systems have seldom been researched. Especially for phages, little is known about their response to antimicrobial exposure. In this study, two identical anoxic-aerobic wastewater treatment systems were established to evaluate the responses of bacterial and phage communities to long-term exposure to antimicrobial agents. The results indicated simultaneous exposure to combined antimicrobials significantly inhibited (p < 0.05) the abundance of phages and bacteria. Metagenomic sequencing analysis indicated the community of bacteria and phages changed greatly at the genus level due to combined antibacterial exposure. Additionally, long-term exposure to antimicrobial agents promoted the attachment of receptor-binding protein genes to Klebsiella, Escherichia and Salmonella (which were all members of Enterobacteriaceae). Compared to that in the control system, the numbers of receptor-binding protein genes on their possible phages (such as Lambdalikevirus and P2likevirus) were also obviously higher when the microorganisms were exposed to antimicrobials. The results are helpful to understanding the microbial communities and tracking the relationship of phage-bacterial host systems, especially under the pressure of antimicrobial exposure.

Endoplasmic reticulum-specific fluorescent probe for the two-photon imaging of endogenous superoxide anion (O2•-) in live cells and zebrafishes.

ER stress has close relation with various metabolic diseases including obesity and insulin resistance, and could result in the abnormal production of ROS including O2-. Real-time and in situ detection of endogenous O2- in ER is vitally important for revealing the physiological roles of O2- during ER stress. Herein, we present an ER-specific two-photon probe (ER-Rs) for the detection of endogenous O2- in living cells and zebrafishes. The probe ER-Rs employed triflate as the response site for O2-, and used p-methylsulfonamide as the ER-specific moiety. In response to O2-, the triflate group of the probe ER-Rs was transformed to hydroxyl and the turn-on fluorescence was produced. The probe ER-Rs displayed highly sensitive and selective response to O2-, and could be employed as an ER-specific two-photon probe for the visualization of endogenous O2- in live cells, tissues and zebrafishes.

Resistance of multidrug resistant Escherichia coli to environmental nanoscale TiO2 and ZnO.

Excessive production and utilization of nanoparticles (NPs) at industrial and household levels releases substantial quantities of NPs into the environment. These can be harmful to different types of organisms and cause adverse effects on ecosystems. Purchased TiO2 and ZnO NPs were characterized via XRD, XPS, FESEM, and Zeta potential. This study elucidates how multidrug resistant Escherichia coli LM13, which was recovered from livestock manure, counteracts the antibacterial activities of TiO2 and ZnO NPs to survive in the environment. E. coli ATCC25922, which is susceptible to antibiotics, was used as control. A dose-response experiment showed that the antibacterial activity of TiO2 was lower than that of ZnO NPs and, LM13 was more resistant to NPs than ATCC25922. An AcrAB-TolC efflux pump along with its regulation genes helped LM13 to minimize NP toxicity. Flow cytometry findings also indicated that the intensity of the side-scatter light parameter increased with TiO2 and ZnO NPs in a dose dependent manner, suggesting NP uptake by the both strains. The generation of reactive oxygen species in LM13 was several-fold lower than in ATCC25922, suggesting that reactive oxygen species mainly contribute to the toxicity mechanism. These results illustrate the necessity to evaluate the impacts of NPs on the survival capacity of bacteria and on the resistance genes in bacteria with higher NP resistance than NP susceptible bacteria.