Landscape of plasmids encoding ß-lactamases in disinfection residual Enterobacteriaceae from wastewater treatment plants.

Conventional disinfection processes, such as chlorination and UV radiation, are ineffective in controling antibiotic-resistant bacteria, especially disinfection residual Enterobacteriaceae (DRE) encoding ß-lactamases, some of which have been classified as "critical priority pathogens" by the World Health Organization. However, few studies have focused on the transferability, phenotype, and genetic characteristics of DRE-derived plasmids encoding ß-lactamases, especially extended-spectrum ß-lactamases and carbapenemases. In this study, we isolated 10 typical DRE harboring plasmid-mediated blaNDM, blaCTX-M, or blaTEM in post-disinfection effluent from two wastewater treatment plants (WWTPs), with transfer frequency ranging from 1.69 × 10-6 to 3.02 × 10-5. According to genomic maps of plasmids, all blaNDM and blaTEM were cascaded with IS26, and blaCTX-M was adjacent to ISEcp1 or IS26, indicating the important role of these elements in the movement of ß-lactamase-encoding genes. The presence of intact class 1 integrons on pWTPN-01 and pWTPC-03 suggested the ability of these DRE-derived plasmids to integrate other exogenous antibiotic resistance genes (ARGs). The coexistence of antibiotic, disinfectant, and heavy metal resistance genes on the same plasmid (e.g., pWTPT-03) implied the facilitating role of disinfectants and heavy metals in the transmission of DRE-derived ARGs. Notably, two plasmid transconjugants exhibited no discernible competitive fitness cost, suggesting a heightened environmental persistence. Furthermore, enhanced virulence induced by ß-lactamase-encoding plasmids in their hosts was confirmed using Galleria mellonella infection models, which might be attributed to plasmid-mediated virulence genes. Overall, this study describes the landscape of ß-lactamase-encoding plasmids in DRE, and highlights the urgent need for advanced control of DRE to keep environmental and ecological security.

Deciphering Multidrug-Resistant Plasmids in Disinfection Residual Bacteria from a Wastewater Treatment Plant.

Current disinfection processes pose an emerging environmental risk due to the ineffective removal of antibiotic-resistant bacteria, especially disinfection residual bacteria (DRB) carrying multidrug-resistant plasmids (MRPs). However, the characteristics of DRB-carried MRPs are poorly understood. In this study, qPCR analysis reveals that the total absolute abundance of four plasmids in postdisinfection effluent decreases by 1.15 log units, while their relative abundance increases by 0.11 copies/cell compared to investigated wastewater treatment plant (WWTP) influent. We obtain three distinctive DRB-carried MRPs (pWWTP-01-03) from postdisinfection effluent, each carrying 9-11 antibiotic-resistant genes (ARGs). pWWTP-01 contains all 11 ARGs within an ∼25 Kbp chimeric genomic island showing strong patterns of recombination with MRPs from foodborne outbreaks and hospitals. Antibiotic-, disinfectant-, and heavy-metal-resistant genes on the same plasmid underscore the potential roles of disinfectants and heavy metals in the coselection of ARGs. Additionally, pWWTP-02 harbors an adhesin-type virulence operon, implying risks of both antibiotic resistance and pathogenicity upon entering environments. Furthermore, some MRPs from DRB are capable of transferring and could confer selective advantages to recipients under environmentally relevant antibiotic pressure. Overall, this study advances our understanding of DRB-carried MRPs and highlights the imminent need to monitor and control wastewater MRPs for environmental security.

Ranking the risk of antibiotic resistance genes by metagenomic and multifactorial analysis in hospital wastewater systems.

Antimicrobial resistance poses a serious threat to human health. Hospital wastewater system (HWS) is an important source of antibiotic resistance genes (ARGs). The risk of ARGs in HWS is still an under-researched area. In this study, we collected publicly metagenomic datasets of 71 hospital wastewater samples from 18 hospitals in 13 cities. A total of 9838 contigs were identified to carry 383 unique ARGs across all samples, of which 2946 contigs were plasmid-like sequences. Concurrently, the primary hosts of ARGs within HWS were found to be Escherichia coli and Klebsiella pneumoniae. To further evaluate the risk of each ARG subtype, we proposed a risk assessment framework based on the importance of corresponding antibiotics as defined by the WHO and three other indicators - ARG abundance (A), mobility (M), and host pathogenicity (P). Ninety ARGs were identified as R1 ARGs having high-risk scores, which meant having a high abundance, high mobility, and carried by pathogens in HWS. Furthermore, 25% to 49% of genomes from critically important pathogens accessed from NCBI carried R1 ARGs. A significantly higher number of R1 ARGs was carried by pathogens in the effluents of municipal wastewater treatment plants from NCBI, highlighting the role of R1 ARGS in accelerating health and environmental risks.

Antibiotic resistance genes alterations in murine guts microbiome are associated with different types of drinking water.

Antibiotic resistance genes (ARGs) are emerging contaminants threatening public health and commonly found in drinking water. However, the effect of different types of drinking water on ARG alterations in the gut microbiome is unclear. This study examines this issue in murine models in three phases (phase I: acclimation using ddH2O; phase II: treatment using different types of water, i.e. river water (RW), tap water (TW) and commercial bottled water (CBW); and phase III: recovery using ddH2O) using high-throughput qPCR and 16S rRNA amplicon sequencing. Results reveal that exposure to different types of drinking water could lead to significant changes in the gut microbiome, mobile genetic elements (MGEs), and ARGs. In phase II, treatment of RW and TW significantly increased the abundance of aminoglycoside and tetracycline resistance genes in mice guts (P < 0.01). In the recovery phase, consuming distilled water was found to restore ARG profiles to a certain extent in mice guts. Procrustes, network, redundancy and variation partitioning analysis indicated that ARG alterations in mice guts might relate to MGEs and bacterial communities. Our work suggests that the type of drinking water consumed may play a crucial role in shaping ARGs in gut microbiomes, emphasizing the urgent need for access to clean drinking water to mitigate the growing threat of antimicrobial resistance.

Comprehensive profiling and risk assessment of antibiotic resistance genes in a drinking water watershed by integrated analysis of air-water-soil.

The prevalence of antibiotic resistance genes (ARGs) in diverse habitats threatens public health. Watersheds represent critical freshwater ecosystems that interact with both the soil and atmosphere. However, a holistic understanding of ARGs distribution across these environmental media is currently inadequate. We profiled ARGs and bacterial communities in air-water-soil in the same watershed area during four seasons using high-throughput qPCR and 16S rRNA gene sequencing. Our findings demonstrated that aminoglycoside resistance genes (58.5%) were dominant in water, and multidrug resistance genes (55.2% and 54.2%) were dominant in soil and air. Five ARGs and nineteen bacterial genera were consistently detected in all samples, were named as shared genes or bacteria. Co-occurrence Network analysis revealed the co-occurrence module of resistance genes, mobile genetic elements (MGEs), and potential bacterial hosts, indicating that shared genes and bacteria may persist and co-spread across different environmental media. The risk assessment framework, based on ARGs' abundance, detection rate, and mobility, identified 33 high-risk ARGs. This is essential to evaluate the health risks of ARGs and to develop strategies to limit the threat of antibiotic resistance. Our study offers new insights into the risks associated with ARGs in the environment and suggests that ARGs may depend on specific bacterial cohabitants that co-exist with MGEs to facilitate their spread across environmental interfaces.

Association between particulate matter (PM)2·5 air pollution and clinical antibiotic resistance: a global analysis.

BACKGROUND: Antibiotic resistance is an increasing global issue, causing millions of deaths worldwide every year. Particulate matter (PM)2·5 has diverse elements of antibiotic resistance that increase its spread after inhalation. However, understanding of the contribution of PM2·5 to global antibiotic resistance is poor. Through univariate and multivariable analysis, we aimed to present the first global estimates of antibiotic resistance and burden of premature deaths attributable to antibiotic resistance resulting from PM2·5 pollution. METHODS: For this global analysis, data on multiple potential predictors (ie, air pollution, antibiotic use, sanitation services, economics, health expenditure, population, education, climate, year, and region) were collected in 116 countries from 2000 to 2018 to estimate the effect of PM2·5 on antibiotic resistance via univariate and multivariable analysis. Data were obtained from ResistanceMap, European Centre for Disease Prevention and Control Surveillance Atlas (antimicrobial-resistance sources), and PLISA Health Information Platform for the Americas. Future global aggregate antibiotic resistance and premature mortality trends derived from PM2·5 in different scenarios (eg, 50% reduced antibiotic use or PM2·5 controlled to 5 µg/m3) were projected until 2050. FINDINGS: The final dataset included more than 11·5 million tested isolates. Raw antibiotic-resistance data included nine pathogens and 43 types of antibiotic agents. Significant correlations between PM2·5 and antibiotic resistance were consistent globally in most antibiotic-resistant bacteria (R2=0·42-0·76, p<0·0001), and correlations have strengthened over time. Antibiotic resistance derived from PM2·5 caused an estimated 0·48 (95% CI 0·34-0·60) million premature deaths and 18·2 (13·4-23·0) million years of life lost in 2018 worldwide, corresponding to an annual welfare loss of US$395 (290-500) billion due to premature deaths. The 5 µg/m3 target of concentration of PM2·5 in the air quality guidelines set by WHO, if reached in 2050, was estimated to reduce antibiotic resistance by 16·8% (95% CI 15·3-18·3) and avoid 23·4% (21·2-25·6) of premature deaths attributable to antibiotic resistance, equivalent to a saving of $640 (580-671) billion. INTERPRETATION: This analysis is the first to describe the association between PM2·5 and clinical antibiotic resistance globally. Results provide new pathways for antibiotic-resistance control from an environmental perspective. FUNDING: National Natural Science Foundation of China, Fundamental Research Funds for the Central Universities, Zhejiang University Global Partnership Fund, and China Postdoctoral Science Foundation.

3D ZnO/Activated Carbon Alginate Beads for the Removal of Antibiotic-Resistant Bacteria and Antibiotic Resistance Genes.

The worldwide prevalence of antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs) have become one of the most urgent issues for public health. Thus, it is critical to explore more sustainable methods with less toxicity for the long-term removal of both ARB and ARGs. In this study, we fabricated a novel material by encapsulating zinc oxide (ZnO) nanoflowers and activated carbon (AC) in an alginate biopolymer. When the dosage of ZnO was 1.0 g (≈2 g/L), the composite beads exhibited higher removal efficiency and a slight release of Zn2+ in water treatment. Fixed bed column experiments demonstrated that ZnO/AC alginate beads had excellent removal capacities. When the flow rate was 1 mL/min, and the initial concentration was 107 CFU/mL, the removal efficiency of ARB was 5.69-log, and the absolute abundance of ARGs was decreased by 2.44-2.74-log. Moreover, the mechanism demonstrated that ZnO significantly caused cell lysis, cytoplasmic leakage, and the increase of reactive oxygen species induced subsequent oxidative stress state. These findings suggested that ZnO/AC alginate beads can be a promising material for removing ARB and ARGs from wastewater with eco-friendly and sustainable properties.

Epidemiological Characteristics and Influencing Factors of Myopia Among Primary School Students in Southern China: A Longitudinal Study.

Objectives: To study the epidemiological characteristics and influencing factors of myopia to provide a scientific basis for the prevention and control of myopia. Methods: 7,597 students studying in grades 1-3 were followed up. Eye examinations and questionnaire surveys were conducted annually from 2019 to 2021. The influencing factors of myopia were analyzed by logistic regression model. Results: The prevalence of myopia among students in grades 1-3 in 2019 was 23.4%, which increased to 41.9% and 51.9% after the 1-and 2-year follow-up, respectively. The incidence of myopia and change in the spherical equivalent refraction (SER) were higher in 2020 than in 2021. The 2-year cumulative incidences of myopia were 2.5%, 10.1%, 15.5%, 36.3%, and 54.1% in students with a baseline SER >+1.50D, +1.00D to +1.50D, +0.50D to +1.00D, 0.00D to +0.50D, and -0.50D to 0.00D, respectively. Outdoor activities, sex, age, baseline SER, parental myopia, sleep time, and digital device exposure were associated with myopia. Conclusion: The prevalence of myopia demonstrated a rapid increase; thus, healthy habits and outdoor activities should be promoted for the prevention and control of myopia.

Microplastics exacerbate co-occurrence and horizontal transfer of antibiotic resistance genes.

Microplastic pollution is a rising environmental issue worldwide. Microplastics can provide a niche for the microbiome, especially for antibiotic-resistant bacteria, which could increase the transmission of antibiotic resistance genes (ARGs). However, the interactions between microplastics and ARGs are still indistinct in environmental settings. Microplastics were found to be significantly correlated with ARGs (p < 0.001), based on the analysis of samples taken from a chicken farm and its surrounding farmlands. Analysis of chicken feces revealed the highest abundance of microplastics (14.9 items/g) and ARGs (6.24 ×108 copies/g), suggesting that chicken farms could be the hotspot for the co-spread of microplastics and ARGs. Conjugative transfer experiments were performed to investigate the effects of microplastic exposure for different concentrations and sizes on the horizontal gene transfer (HGT) of ARGs between bacteria. Results showed that the microplastics significantly enhanced the bacterial conjugative transfer frequency by 1.4-1.7 folds indicating that microplastics could aggravate ARG dissemination in the environment. Potential mechanisms related to the up-regulation of rpoS, ompA, ompC, ompF, trbBp, traF, trfAp, traJ, and down-regulation of korA, korB, and trbA were induced by microplastics. These findings highlighted the co-occurrence of microplastics and ARGs in the agricultural environment and the exacerbation of ARGs' prevalence via rising the HGT derived from microplastics.

Deciphering basic and key traits of antibiotic resistome in influent and effluent of hospital wastewater treatment systems.

Hospital wastewater treatment system (HWTS) is an important source and environmental reservoir of clinically relevant antibiotic resistance genes (ARGs). However, how antibiotic resistome of clinical wastewater changed in HWTS is poorly understood. Herein, the basic quantitative traits (i.e., diversity and abundance) of ARGs in three HWTSs were profiled by metagenomics. In total, 709 ARG subtypes belonging to 20 ARG types were detected with relative abundance ranging from 1.12 × 10-5 to 7.33 × 10-1 copies/cell. Notably, most ARGs could not be significantly removed by chlorination treatment in the HWTS. These ARGs were identified to confer resistance to almost all major classes of antibiotics and include ARGs of last-resort antibiotics, such as blaNDM, mcr and tet(X) which were abundantly occurred in HWTS with 19, 5 and 7 variants, respectively. Moreover, qualitative analysis based on metagenome-assembled genome (MAG) analysis revealed that the putative hosts of the identified ARGs were broadly distributed into at least 8 dominant bacterial phyla. Of the 107 ARG-carrying MAGs recovered, 39 encoded multi-antibiotic resistance and 16 belonged to antibiotic resistant pathogens. Further analysis of co-occurrence patterns of ARGs with mobile genetic elements suggested their potential mobility. These key qualitative traits of ARGs provided further information about their phylogeny and genetic context. This study sheds light on the key traits of ARGs associated with resistance dissemination and pathogenicity and health risks of clinical wastewater.

Research trends and hotspots in the relationship between outdoor activities and myopia: A bibliometric analysis based on the web of science database from 2006 to 2021.

Objectives: This study aimed to explore the current status, hotspots, and emerging research trends regarding the relationship between outdoor activities and myopia. Methods: Publications on the relationship between outdoor activities and myopia from 2006 to 2021 were collected from the Web of Science Core Collection database. CiteSpace (version 6.1.R2) was used to performed a bibliometric analysis, and R software (version 4.1.0) was used to visualize the trends and hot map of publications. Results: A total of 640 publications were collected and analyzed in the present study. China was the major contributor (n = 204), followed by the United States of America (n = 181) and Australia (n = 137). The United States of America had the most extensive foreign cooperation (centrality = 0.25), followed by Australia (centrality = 0.20). The National University of Singapore contributed the largest number of publications (n = 48), followed by Sun Yat-Sen University (n = 41) and the Australian National University (n = 41). Among institutions, Cardiff University in the United Kingdom had the most extensive foreign cooperation (centrality = 0.12), followed by the National University of Singapore (centrality = 0.11). Saw S from Singapore had the largest number of publications (n = 39), followed by Morgan I from Australia (n = 27) and Jonas J from Germany (n = 23). Investigative ophthalmology & visual science is the most important journal to study the relationship between outdoor activities and myopia. "Global Prevalence of Myopia and High Myopia and Temporal Trends from 2000 through 2050" published by Holden BA was the most cited paper in this field with 177 citations. Co-occurrence and burst analyses of keywords showed that research trends and hotspots in this field focused mainly on "risk," "prevention" and "school". Conclusions: The influence of outdoor activities on myopia remains a concern. In the future, deeper cooperation between countries or institutions is required to explore the effects of outdoor activities on myopia. Outdoor activities for the prevention of myopia and reduction of the risk of myopia among school students may be the focus of future research.

The accuracy of the axial length and axial length/corneal radius ratio for myopia assessment among Chinese children.

Objectives: The aim of this study was to assess the association between axial length/corneal radius ratio (AL/CR ratio), AL, and refractive status and evaluate the accuracy of AL and AL/CR ratio for myopia assessment among Chinese children. Methods: A diagnostic trial was conducted in Shenzhen Eye Hospital from June 2020 to December 2020. Cycloplegic refraction and demographic characteristic survey were carried out, and AL and CR were measured. The Pearson correlation analysis between AL, AL/CR ratio, and spherical equivalent (SE) was carried out. The sensitivity, specificity, Youden index, positive predictive value, and negative predictive value of the AL/CR ratio and AL for myopia assessment were analyzed using cycloplegic refraction as the gold standard by drawing receiver operating characteristic (ROC) curves. Results: A total of 300 children aged 8-18 years participated in this study. The Pearson correlation coefficient between AL and SE was -0.667 (P < 0.05) and -0.754 (P < 0.05) between AL/CR ratio and SE. There were significant differences in SE, AL, and AL/CR ratio among different age groups (p < 0.05). SE decreased by 1.185 diopter (D) for every 1 mm increase in AL and decreased by 0.667 D for every 0.1 increase in the AL/CR ratio. Taking cycloplegic refraction SE ≤ -0.50 D as the gold standard for the diagnosis of myopia, the area under the ROC curve of AL for myopia assessment was 0.836 (95% confidence interval [CI]: 0.767-0.906), with specificity, sensitivity, and Youden index of 0.833, 0.767, and 0.600, respectively. The area under the ROC curve of AL/CR ratio for myopia assessment was 0.937 (95% CI: 0.878-0.996), with specificity, sensitivity, Youden index, positive predictive value, and negative predictive value of 0.703, 0.913, 0.622, 0.956, and 0.771, respectively. The area under the ROC curve of the combination of AL/CR ratio and parental myopia for myopia assessment was 0.976 (95% CI: 0.957-0.996). Conclusion: The correlation between SE and AL/CR ratio was stronger than that between SE and AL in children. The AL/CR ratio may be an alternative indicator for myopia assessment in children, and the combination of demographic factors and AL/CR ratio can improve the accuracy of myopia assessment.

Short-term inhalation exposure evaluations of airborne antibiotic resistance genes in environments.

Antibiotic resistance is a sword of Damocles that hangs over humans. In regards to airborne antibiotic resistance genes (AARGs), critical knowledge gaps still exist in the identification of hotspots and quantification of exposure levels in different environments. Here, we have studied the profiles of AARGs, mobile genetic elements (MGEs) and bacterial communities in various atmospheric environments by high throughput qPCR and 16S rRNA gene sequencing. We propose a new AARGs exposure dose calculation that uses short-term inhalation (STI). Swine farms and hospitals were high-risk areas where AARGs standardised abundance was more abundant than suburbs and urban areas. Additionally, resistance gene abundance in swine farm worker sputum was higher than that in healthy individuals in other environments. The correlation between AARGs with MGEs and bacteria was strong in suburbs but weak in livestock farms and hospitals. STI exposure analysis revealed that occupational intake of AARGs (via PM10) in swine farms and hospitals were 110 and 29 times higher than in suburbs, were 1.5 × 104, 5.6 × 104 and 5.1 × 102 copies, i.e., 61.9%, 75.1% and 10.7% of the overall daily inhalation intake, respectively. Our study comprehensively compares environmental differences in AARGs to identify high-risk areas, and forwardly proposes the STI exposure dose of AARGs to guide risk assessment.

Trends in Myopia Development Among Primary and Secondary School Students During the COVID-19 Pandemic: A Large-Scale Cross-Sectional Study.

Objectives: To evaluate myopia development among primary and secondary school students during the coronavirus 2019 (COVID-19) pandemic. Methods: A cross-sectional study was conducted to evaluate the development of myopia among students in Shenzhen, China during the COVID-19 outbreak. Results: The study included 1,472,957 and 1,573,824 students in 2019 and 2020, respectively. The prevalence of myopia was 46.9 and 50.5% in 2019 and 2020, respectively. The prevalence of myopia among students in the former Shenzhen Special Economic Zone (SEZ) was higher than that in areas outside the former Shenzhen SEZ (2019: 47.0 vs. 43.7%; 2020: 50.5 vs. 47.3%). The prevalence of myopia among girls was higher than that among boys (2019: 50.4 vs. 44.0%; 2020: 54.0 vs. 47.6%). The 50th percentile (P50) of spherical equivalent refraction (SER) in the right eye among girls was lower than that in boys. The prevalence of myopia continued to increase as the grade increased, with the greatest annual increase observed in Grades 2-5 (3.4-3.9%). The P50 of SER in the right eye of students decreased as the grade increased. Conclusions: The prevalence of myopia among students increased during the COVID-19 pandemic, especially in primary school Grades 2-5.

Mechanism and potential risk of antibiotic resistant bacteria carrying last resort antibiotic resistance genes under electrochemical treatment.

The significant rise in the number of antibiotic resistance genes (ARGs) that resulted from our abuse of antibiotics could do severe harm to public health as well as to the environment. We investigated removal efficiency and removal mechanism of electrochemical (EC) treatment based on 6 different bacteria isolated from hospital wastewater carrying 3 last resort ARGs including NDM-1, mcr-1 and tetX respectively. We found that the removal efficiency of ARGs increased with the increase of both voltage and electrolysis time while the maximum removal efficiency can reach 90%. The optimal treatment voltage and treatment time were 3 V and 120 min, respectively. Temperature, pH and other factors had little influence on the EC treatment process. The mechanism of EC treatment was explored from the macroscopic and microscopic levels by scanning electron microscopy (SEM) and flow cytometry. Our results showed that EC treatment significantly changed the permeability of cell membrane and caused cells successively experience early cell apoptosis, late cell apoptosis and cell necrosis. Moreover, compared with traditional disinfection methods, EC treatment had less potential risks. The conjugative transfer frequencies of cells were significantly reduced after treatment. Less than 1% of bacteria entered the viable but nonculturable (VBNC) state and less than 5% of intracellular ARGs (iARGs) turned into extracellular ARGs (eARGs). Our findings provide new insights into as well as important reference for future electrochemical treatment in removing ARB from hospital wastewater.

Mechanisms underlying the effect of chlorination and UV disinfection on VBNC state Escherichia coli isolated from hospital wastewater.

The occurrence of viable but non-culturable (VBNC) bacteria in the wastewater system poses a huge threat to environmental and public health, in particular in hospital wastewater treatment system (HWTS). HWTS-oriented studies have been conducted to assess the effectiveness of chlorination and UV disinfection using indigenous bacteria. Results revealed that the VBNC Escherichia coli and ARGs remained persistent even at high chlorination (12 mg/L for 2.5 h) and UV doses (1000 mJ/cm2). The molecular mechanisms underlying chlorination-/UV-induced VBNC state in E. coli were explored through the transcriptomics and results suggested that most energy-dependent physiological activities (e.g., metabolism) have been suppressed in VBNC E. coli, while the pathogenicity-related genes varied insignificantly compared to the culturable cells, indicating that the VBNC E. coli could potentially display pathogenicity. Further Galleria mellonella model experiment has confirmed that although the disinfection-induced VBNC state made cells less infectious, these cells could regain their pathogenicity after resuscitation. This in vitro study can be used as a reference for studies on infections from VBNC bacteria and highlights the health risk due to VBNC pathogens in hospital effluents. There is a need to develop effluent standards specifically for healthcare facilities, and a stricter downstream disinfection strategy should be considered for the removal of VBNC cells and ARGs in the effluent.

Landscape of genes in hospital wastewater breaking through the defense line of last-resort antibiotics.

Hospital wastewater contains abundant antibiotics, antibiotic resistance genes (ARGs), and pathogens. Last-resort antibiotic resistance genes (LARGs) include the New Delhi metallo-ß-lactamase gene blaNDM, mobile colistin resistance gene mcr and tigecycline resistance gene tet(X) which confers resistance to carbapenems, colistin and tigecycline. The presence and significance of LARGs in hospital wastewater treatment systems (HWTS) have not yet been systematically explored. Here, LARG variants were shown to be prevalent both influents and effluents of HWTS. A total of 989 Enterobacteriaceae isolates that confer resistance to last-resort antibiotics were collected from effluents and multiple genetic contexts of LARGs were analyzed. LARGs-carrying plasmids were confirmed to show high multidrug phenotypes and transferability. We also discovered the co-occurrence of plasmids harboring blaNDM-1 and mcr-1 in single Escherichia coli, as well as E. coli HM016 containing two unique mcr-1-carrying plasmids. This result might accelerate co-dissemination of LARGs under environmental selection pressure. Different core genetic arrangements in these strains suggest several evolutionary pathways in HWTS. The resistance functions of LARGs were confirmed in vitro and in vivo by mass spectrometry. This study provides novel insights into the diversity, genetic context and function of critical ARGs in HWTS. The results raise the concern that LARGs may further spread into the environment, thus, more stringent discharge standards and regulations for hospital wastewater are urgently needed.

Dissemination of antibiotic resistance genes in swimming pools and implication for human skin.

Swimming pools are crowd-gathering places that are associated with numerous outbreaks of water-borne diseases. Herein, we investigated the distribution of antibiotic resistance genes (ARGs) and bacterial communities in swimming pools and determined the influencing factors and potential human exposure. Sixteen swimming pools with different bather loads (0.01-0.16 person/m2·h) were investigated. Water samples were collected, before opening and after closing of the facilities, from six swimming pools, and skin samples were collected from volunteers. Comprehensive approaches, high-throughput qPCR and 16S rRNA gene sequencing, were used. The results showed that swimming pools contained a higher relative abundance (0.62 gene copies/16S rRNA) and absolute abundance (6.57×108 gene copies/L) of ARGs on average. Bather loads contributed to the increase of core ARGs, and the absolute abundance of ARGs significantly increased by 1.47-1.94 orders of magnitude when the bather load was more than 0.1 person/m2·h. Dermal contact was estimated as the main exposure route of ARGs. Eighteen ARGs that were not detected before swimming were found on human skin and remained after showering. Furthermore, the event intake burden of ARGs via dermal contact was higher than that via ingestion when swimming. This study provides an assessment of ARGs and antibiotic-resistant bacteria (ARB) in swimming pools and helps to define the health risks to swimmers.

Spread of antibiotic resistance genes and microbiota in airborne particulate matter, dust, and human airways in the urban hospital.

Antimicrobial resistance is an increasingly serious threat to public health worldwide. The presence of antibiotic resistance genes (ARGs) in human airways and relevant environments has not received significant attention. In this study, abundances of ARGs and microbes from airborne particulate matter, dust, and human airways in a hospital were profiled using high-throughput qPCR and 16S rRNA gene sequencing. More diverse ARGs and microbes in indoor dust and higher levels of ARGs in particulate matter PM10 and PM2.5 were observed. Macrolides and aminoglycoside resistance genes were the most abundant ARGs in the airway and environmental samples, respectively. Moreover, the co-occurrences of priority pathogens, ARGs, and mobile genetic elements (MGEs) were shown by the Network analysis. Campylobacter spp. and Staphylococcus spp. positively correlated with fluoroquinolone (vatC-02, mexD) and ß-lactams (blaZ, mecA) resistance genes, respectively. In this regard, based on SourceTracker analysis, inhalable particles contributed to 4.0% to 5.5% of ARGs in human airway samples, suggesting an important exchange between airborne inhalable particles and human commensals. This study may advance knowledge about ARGs in airborne particulate matter and dust associated environments, reveal their potential link between environments and humans, and provide a new sight and fundamental data for ARG risk assessment.

Temporal variation and sharing of antibiotic resistance genes between water and wild fish gut in a peri-urban river.

Antibiotic resistance genes (ARGs) as emergence contaminations have spread widely in the water environment. Wild fish may be recipients and communicators of ARGs in the water environment, however, the distribution and transmission of ARGs in the wild fish and relevant water environment were rarely reported. Here, we have profiled ARGs and bacterial communities in wild freshwater fish and relevant water in a peri-urban river using high-throughput qPCR and 16S rRNA gene sequence. A total of 80 and 220 unique ARG subtypes were identified in fish and water samples. Fish and water both showed significant ARG seasonal variations (P < 0.05). The highest absolute abundance of ARGs in fish and water occurred in summer (1.32 × 109 copies per g, on average) and autumn (9.04 × 106 copies per mL), respectively. In addition, the bipartite network analysis showed that 9 ARGs and 1 mobile genetic element continuously shared in fish and water. Furthermore, bacteria shared in fish and water were found to significantly correlate with shard ARGs. The findings demonstrate that bacteria and ARGs in fish and water could interconnect and ARGs might transfer between fish and water using bacteria as a spreading medium.