Airborne antibiotic resistome and microbiome in pharmaceutical factories.

Antimicrobial resistance is considered to be one of the biggest public health problems, and airborne transmission is an important but under-appreciated pathway for the spread of antibiotic resistance genes (ARGs) in the environment. Previous research has shown pharmaceutical factories to be a major source of ARGs and antibiotic resistant bacteria (ARB) in the surrounding receiving water and soil environments. Pharmaceutical factories are hotspots of antibiotic resistance, but the atmospheric transmission and its environmental risk remain more concerns. Here, we conducted a metagenomic investigation into the airborne microbiome and resistome in three pharmaceutical factories in China. Soil (average: 38.45%) and wastewater (average: 28.53%) were major contributors of airborne resistome. ARGs (vanR/vanS, blaOXA, and CfxA) conferring resistance to critically important clinically used antibiotics were identified in the air samples. The wastewater treatment area had significantly higher relative abundances of ARGs (average: 0.64 copies/16S rRNA). Approximately 28.2% of the detected airborne ARGs were found to be associated with plasmids, and this increased to about 50% in the wastewater treatment area. We have compiled a list of high-risk airborne ARGs found in pharmaceutical factories. Moreover, A total of 1,043 viral operational taxonomic units were identified and linked to 47 family-group taxa. Different CRISPR-Cas immune systems have been identified in bacterial hosts in response to phage infection. Similarly, higher phage abundance (average: 2451.70 PPM) was found in the air of the wastewater treatment area. Our data provide insights into the antibiotic resistance gene profiles and microbiome (bacterial and non-bacterial) in pharmaceutical factories and reveal the potential role of horizontal transfer in the spread of airborne ARGs, with implications for human and animal health.

Antibiotic resistome in landfill leachate and impact on groundwater.

Landfill leachate is a hotspot in antibiotic resistance development. However, little is known about antibiotic resistome and host pathogens in leachate and their effects on surrounding groundwater. Here, metagenomic sequencing was used to explore profiles, host bacteria, environmental risks and influencing factors of antibiotic resistome in raw and treated leachate and surrounding groundwater of three landfills. Results showed detection of a total of 324 antibiotic resistance genes (ARGs). The ARGs conferring resistance to multidrug (8.8 %-25.7 %), aminoglycoside (13.1 %-39.2 %), sulfonamide (10.0 %-20.9 %), tetracycline (5.7 %-34.4 %) and macrolide-lincosamide-streptogramin (MLS, 5.3 %-29.5 %) were dominant in raw leachate, while multidrug resistance genes were the major ARGs in treated leachate (64.1 %-83.0 %) and groundwater (28.7 %-76.6 %). Source tracking analysis suggests non-negligible influence of leachate on the ARGs in groundwater. The pathogens including Acinetobacter pittii, Pseudomonas stutzeri and P. alcaligenes were the major ARG-carrying hosts. Variance partitioning analysis indicates that the microbial community, abiotic variables and their interaction contributed most to the antibiotic resistance development. Our results shed light on the dissemination and driving mechanisms of ARGs from leachate to the groundwater, indicating that a comprehensive risk assessment and efficient treatment approaches are needed to deal with ARGs in landfill leachate and nearby groundwater. ENVIRONMENTAL IMPLICATIONS: Antibiotic resistance genes are found abundant in the landfill sites, and these genes could be disseminated into groundwater via leaching of wastewater and infiltration of leachate. This results in deterioration of groundwater quality and human health risks posed by these ARGs and related pathogens. Thus measures should be taken to minimize potential negative impacts of landfills on the surrounding environment.

Swine farming shifted the gut antibiotic resistome of local people.

Antibiotic resistance genes (ARGs) are prevalent in the livestock environment, but little is known about impacts of animal farming on the gut antibiotic resistome of local people. Here we conducted metagenomic sequencing to investigate gut microbiome and resistome of residents in a swine farming village as well as environmental relevance by comparing with a nearby non-farming village. Results showed a shift of gut microbiome towards unhealthy status in the residents of swine farming village, with an increased abundance and diversity in pathogens and ARGs. The resistome composition in human guts was more similar with that in swine feces and air than that in soil and water. Mobile gene elements were closely associated with the prevalence of gut resistome. Some plasmid-borne ARGs were colocalized in similar genetic contexts in gut and environmental samples. Metagenomic binning obtained 47 ARGs-carrying families in human guts, and therein Enterobacteriaceae posed the highest threats in antibiotic resistance and virulence. Several ARGs-carrying families were shared by gut and environmental samples (mainly in swine feces and air), and the ARGs were evolutionarily conservative within genera. The findings highlight that swine farming can shape gut resistome of local people with close linkage to farm environmental exposures.

Removal, fate, and bioavailability of fluoroquinolone antibiotics in a phytoremediation system with four wetland plants: Combing dynamic DGT and traditional methods.

Phytoremediation is considered an effective technology for remediating antibiotic-contaminated water; however, its underlying mechanisms remain poorly understood. Therefore, this study investigated the phytoremediation potential of fluoroquinolone antibiotics (FQs) by different wetland plant species. The phytoremediation rates of ΣFQs were 46-69 %, and rhizosphere microorganism degradation (accounting for 90-93 %) dominated the FQ removal over that of plant uptake and hydrolysis. Dissipation of the FQs in the hydroponic system followed a first-order kinetic model. The joint action of the more powerful absorptive capacity of plants and stronger microbial degradation ability in the rhizosphere was the reason that Cyperus papyrus showed significantly higher FQ phytoremediation rates than the other three plant species, which implied that the plant species is a critical factor affecting phytoremediation efficiency. The FQ distribution in plant tissues decreased from root > stem > leaf, suggesting that FQs were more concentrated in the roots than in the aboveground tissues. Negative correlations between the diffusive gradient in thin films and root concentrations implied that these wetland plant species took up FQs mainly via active transport mechanism (requiring some vectors, perhaps via exudates); whereas, the process of root-to-stem transfer and upward transport represented passive transport, which mainly depended on transpiration. These results facilitate an improved understanding of phytoremediation processes and improve their future applications.

Swine farm groundwater is a hidden hotspot for antibiotic-resistant pathogenic Acinetobacter.

Acinetobacter is present in the livestock environment, but little is known about their antibiotic resistance and pathogenic species in the farm groundwater. Here we investigated antibiotic resistance of Acinetobacter in the swine farm groundwater (JZPG) and residential groundwater (JZG) of a swine farming village, in comparison to a nearby (3.5 km) non-farming village (WTG) using metagenomic and culture-based approaches. Results showed that the abundance of antibiotic resistome in some JZG and all JZPG (~3.4 copies/16S rRNA gene) was higher than that in WTG (~0.7 copies/16S rRNA gene), indicating the influence of farming activities on both groundwater types. Acinetobacter accounted for ~95.7% of the bacteria in JZG and JZPG, but only ~8.0% in WTG. They were potential hosts of ~95.6% of the resistome in farm affected groundwater, which includes 99 ARG subtypes against 23 antibiotic classes. These ARGs were associated with diverse intrinsic and acquired resistance mechanisms, and the predominant ARGs were tetracyclines and fluoroquinolones resistance genes. Metagenomic binning analysis elucidated that non-baumannii Acinetobacter including A. oleivorans, A. beijerinckii, A. seifertii, A. bereziniae and A. modestus might pose environmental risks because of multidrug resistance, pathogenicity and massive existence in the groundwater. Antibiotic susceptibility tests showed that the isolated strains were resistant to multiple antibiotics including sulfamethoxazole (resistance ratio: 96.2%), levofloxacin (42.5%), gatifloxacin (39.0%), ciprofloxacin (32.6%), tetracycline (32.0%), doxycycline (29.0%) and ampicillin (12.0%) as well as last-resort polymyxin B (31.7%), colistin (24.1%) and tigecycline (4.1%). The findings highlight potential prevalence of groundwater-borne antibiotic-resistant pathogenic Acinetobacter in the livestock environment.

Mariculture affects antibiotic resistome and microbiome in the coastal environment.

Antibiotics are increasingly used and released into the marine environment due to the rapid development of mariculture, resulting in spread of antibiotic resistance. The pollution, distribution, and characteristics of antibiotics, antibiotic resistance genes (ARGs) and microbiomes have been investigated in this study. Results showed that 20 antibiotics were detected in Chinese coastal environment, with predominance of erythromycin-H2O, enrofloxacin and oxytetracycline. In coastal mariculture sites, antibiotic concentrations were significantly higher than in control sites, and more types of antibiotics were detected in the South than in the North of China. Residues of enrofloxacin, ciprofloxacin and sulfadiazine posed high resistance selection risks. ß-Lactam, multi-drug and tetracycline resistance genes were frequently detected with significantly higher abundance in the mariculture sites. Of the 262 detected ARGs, 10, 26, and 19 were ranked as high-risk, current-risk, future-risk, respectively. The main bacterial phyla were Proteobacteria and Bacteroidetes, of which 25 genera were zoonotic pathogens, with Arcobacter and Vibrio in particular ranking in the top10. Opportunistic pathogens were more widely distributed in the northern mariculture sites. Phyla of Proteobacteria and Bacteroidetes were the potential hosts of high-risk ARGs, while the conditional pathogens were associated with future-risk ARGs, indicating a potential threat to human health.

Airborne antibiotic resistome and human health risk in railway stations during COVID-19 pandemic.

Antimicrobial resistance is recognized as one of the greatest public health concerns. It is becoming an increasingly threat during the COVID-19 pandemic due to increasing usage of antimicrobials, such as antibiotics and disinfectants, in healthcare facilities or public spaces. To explore the characteristics of airborne antibiotic resistome in public transport systems, we assessed distribution and health risks of airborne antibiotic resistome and microbiome in railway stations before and after the pandemic outbreak by culture-independent and culture-dependent metagenomic analysis. Results showed that the diversity of airborne antibiotic resistance genes (ARGs) decreased following the pandemic, while the relative abundance of core ARGs increased. A total of 159 horizontally acquired ARGs, predominantly confering resistance to macrolides and aminoglycosides, were identified in the airborne bacteria and dust samples. Meanwhile, the abundance of horizontally acquired ARGs hosted by pathogens increased during the pandemic. A bloom of clinically important antibiotic (tigecycline and meropenem) resistant bacteria was found following the pandemic outbreak. 251 high-quality metagenome-assembled genomes (MAGs) were recovered from 27 metagenomes, and 86 genera and 125 species were classified. Relative abundance of ARG-carrying MAGs, taxonomically assigned to genus of Bacillus, Pseudomonas, Acinetobacter, and Staphylococcus, was found increased during the pandemic. Bayesian source tracking estimated that human skin and anthropogenic activities were presumptive resistome sources for the public transit air. Moreover, risk assessment based on resistome and microbiome data revealed elevated airborne health risks during the pandemic.

Airborne bacterial community and antibiotic resistome in the swine farming environment: Metagenomic insights into livestock relevance, pathogen hosts and public risks.

Globally extensive use of antibiotics has accelerated antimicrobial resistance (AMR) in the environment. As one of the biggest antibiotic consumers, livestock farms are hotspots in AMR prevalence, especially those in the atmosphere can transmit over long distances and pose inhalation risks to the public. Here, we collected total suspended particulates in swine farms and ambient air of an intensive swine farming area. Bacterial communities and antibiotic resistomes were analyzed using amplicon and metagenomic sequencing approaches. AMR risks and inhalation exposure to potential human-pathogenic antibiotic-resistant bacteria (HPARB) were subsequently estimated with comparison to the reported hospital samples. The results show that swine farms shaped the airborne bacterial community by increasing abundances, reducing diversities and shifting compositions. Swine feces contributed 77% of bacteria to swine farm air, and about 35% to ambient air. Airborne antibiotic resistomes in swine farms mainly conferred resistance to tetracyclines, aminoglycosides and lincosamides, and over 48% were originated from swine feces. Distinct to the hospital air, Firmicutes were dominant bacteria in swine farming environments with conditional pathogens including Clostridium, Streptococcus and Aerococcus being major hosts of antibiotic resistance genes (ARGs). Therein, genomes of S. alactolyticus carrying (transposase/recombinase-associated) ARGs and virulence factor genes were retrieved from the metagenomes of all swine feces and swine farm air samples, but they were not detected in any hospital air samples. This suggests the indication of S. alactolyticus in swine farming environments with potential hazards to human health. Swine farm air faced higher AMR risks than hospital air and swine feces. The inhalation intake of HPARB by a swine farm worker was about three orders of magnitude higher than a person who works in the hospital. Consequently, this study depicted atmospheric transmission of bacteria and antibiotic resistomes from swine feces to the environment.

The variations of antibiotics and antibiotic resistance genes in two subtropical large river basins of south China: Anthropogenic impacts and environmental risks.

Emission of antibiotics into riverine environments affects aquatic ecosystem functions and leads to the development of antibiotic resistance. Here, the profiles of forty-four antibiotics and eighteen antibiotic resistance genes (ARGs) were analyzed in two large rivers of the Pearl River System. In addition, the risks of ecotoxicity and resistance selection posed by the antibiotics were estimated. As compared to the reservoirs, the river sections close to the urban and livestock areas contained more antibiotics and ARGs. Seasonal variations of antibiotics (higher in the dry season) and relative ARGs (normalized by 16S rRNA gene, higher in the wet season) were found in the water, but not in the sediment. Sulfonamide resistance genes were the most prevalent ARGs in both river water and sediment. Antibiotic concentration was correlated with ARG abundance in the water, indicating that antibiotics play a critical role in ARG spread. In addition, oxytetracycline was the most abundant antibiotic with concentrations up to 2030 ng/L in the water and 2100 ng/g in the sediment respectively, and posed the highest risks for resistance selection. Oxytetracycline, tetracycline and sulfamethoxazole were expected to be more ecotoxicologically harmful to aquatic organisms, while ofloxacin, enrofloxacin, norfloxacin, chlortetracycline, oxytetracycline and tetracycline posed ecotoxicological risks in the sediment. The Nanliujiang river with intensive livestock activities was contaminated by antibiotics and ARGs and faced high ecotoxicological and resistance selection risks. Collectively, these findings reflect the impacts of anthropogenic activities on the spread of antibiotic resistance in large river basins.

Rapid target and non-target screening method for determination of emerging organic chemicals in fish.

An effective analytical method for target and non-target screening of multi-class emerging organic chemicals in aquaculture fish muscle samples using liquid chromatography coupled with high resolution time-of-flight mass spectrometry (LC-QTOF-MS/MS) was reported. Two sample pretreatment methods (QuEChERS and Captiva EMR-Lipid cartridge) were compared and assessed for 151 organic compounds covering a wide range of log Kow (-1.37-11.51) in freeze-dried fish samples. Captiva EMR-Lipid cartridge outperformed QuEChERS by high detection frequency (> 87.5%) and effective matrix removal with satisfactory standard deviation of the compounds (< 20%), and meanwhile provided acceptable recoveries for most organic chemicals at three spiking concentration levels (10, 50, and 100 ng g-1). Spiking experiments suggested that the developed non-target screening workflow showed further convincible identification results with over 83.3% of the chemicals confirmed at even low spiking levels. A list of 867 organic chemicals were tentatively identified in fish samples collected from fish ponds with 21 of them classified as high confidence levels (L1 and L2). The results showed that the simplified analytical strategy can be applied for the quantification and screening analysis of a broad range of emerging organic chemicals in fish samples.

Anthropogenic activities and seasonal properties jointly drive the assemblage of bacterial communities in subtropical river basins.

Anthropogenic activities have inevitably impacted riverine ecosystems, yet their overall contribution to the assemblage of bacterial communities at a large river basin scale remains unclear. In this study, 16S amplicon sequencing was implemented to investigate the bacterial ecosystems in paired water and sediment of North River and West River basins in South China., which contains various anthropogenic environments (e.g., rural/urban area, mining area and livestock area). Subsequently, the links between bacterial community and various types of emerging pollutants in river water were analyzed. The results show that the bacterial assemblage of water and sediment had their own properties that the bacterial community of sediment were mainly affected by seasonal properties, while the bacterial community of water were affected by both seasons and anthropogenic activities. Therein, the aquatic bacterial compositions and abundances were driven by changes in temperature, dissolved oxygen and the emerging pollutants. The dominant phyla Proteobacteria and Firmicutes exhibited adaptability to the mining-affected regions, therein many clades (e.g., Beijerinckiaceae, Acetobacteraceae and Mycobacteriaceae) were also prevalent in the livestock-affected and densely-populated regions. In addition, these two phyla presented associations to the antibiotic resistance in water. The levels of antibiotics, relative antibiotic resistance gens (ARGs) and non-antibiotic pharmaceuticals (NAPs) were closely related to bacterial community composition, diversity and functional diversity, indicating their drive in shifting bacterial communities. Collectively, this work provides a basis for understanding the contribution of anthropogenic activities in shifting bacterial community at a large river basin scale. Further, the results provide new insights for expansion of ecological assessment.

Antibiotics, antibiotic resistance genes and microbial community in grouper mariculture.

Increasing use of feed and medicine in mariculture could cause negative environmental impacts such as habitat modification, microbial disease development and antibiotic resistance. Here we investigated contamination of antibiotics and antibiotic resistance genes (ARGs), and composition of microbial community in grouper mariculture systems in Hainan province, China. Results showed detection of various antibiotic residues with the dominance of fluoroquinolones and tetracyclines in the six grouper cultivation systems. The concentrations of the detected antibiotics in the grouper mariculture water were significantly higher than those in the original seawater. Some of the detected antibiotics such as enrofloxacin, ciprofloxacin, ofloxacin, oxytetracycline and erythromycin in the mariculture water and/or sediment would pose high resistance selection risks. Sulfonamides resistance genes sul1 and sul2 were found to be predominant in water and sediment, while tetracycline resistance genes were prevalent in fish gill and gut. The dominant bacterial phyla in water and sediments were Bacteroides, Actinomycetes, and Proteobacteria, while the dominant ones in fish gill and gut were the Proteobacteria. Genera of Vibrio and Mycobacterium in the core microbiota were important zoonotic pathogens, and there was a significant positive correlation between Vibrio and ARGs. Phyla of Proteobacteria, Actinomyces, and Cyanobacteria were positively correlated to ARGs, indicating that these microorganisms are potential hosts of ARGs. The putative functions of microbiome related to antibiotic resistance and human diseases were significantly higher in fish than in the mariculture environment. This study suggests that mariculture system is a reservoir of ARGs, and the use of antibiotics in mariculture could induce the increase of antibiotic resistance and the prevalence of opportunistic pathogens.

The fate of sulfonamides in the process of phytoremediation in hydroponics.

Phytoremediation has been proven to be an alternative in-situ treatment technique for sulfonamide polluted wastewater. However, the fate of sulfonamides in the phytoremediation process of multiple sulfonamides coexistence is unclear. Therefore, the possibility and mechanism of phytoremediation of ten sulfonamides by different wetland plants through hydroponics were investigated in this study. The phytoremediation rates of Σsulfonamides by different wetland plants were from 44.5% to 56.9%. Mass balance analysis showed that rhizosphere biodegradation (90.2% - 92.2%) dominated the phytoremediation of Σsulfonamides, while hydrolysis (7.63% - 8.95%) and plant uptake (0.05% - 0.17%) accounted for a small proportion. It is worth mentioning that the dissipation of the target sulfonamides in the hydroponic system followed the first-order reaction kinetic model, with half-lives of 13.3 d to 53.3 d, which are close to or even lower than that of aerobic biodegradation in river water, sediment, and piggery wastewater. Six of the ten spiked sulfonamides were detected in plant samples demonstrated that the selective uptake of plants under the coexistence of multiple sulfonamides. The distribution of sulfonamides (concentrations and uptake amounts) in plant tissues followed the sequence of root > stem > leaf in this study, but the distribution in stems and leaves needs further study. The uptake and rhizosphere biodegradation of Cyperus papyrus to sulfonamides are optimally resulting that its phytoremediation rate is significantly higher than other plants (p < 0.05), which indicates that plant species is one of the key factors affecting the phytoremediation efficiency of sulfonamides. These findings verify the feasibility of phytoremediation of sulfonamides, and provide new insights into the fate of sulfonamides in the process of phytoremediation.

Antibiotic resistance genes in surface water and groundwater from mining affected environments.

Mining activities are known to generate a large amount of mine tailings and acid mine drainage which contain varieties of heavy metals. Heavy metals play an important role in co-selection for bacterial antibiotic resistance. However, the characteristics of antibiotic resistance genes (ARGs) in mining-affected water environments are still unclear. Here we investigated the pollution of metals, profiles of ARGs, mobile genetic elements (MGEs) and microbial community in mining-affected surface water and groundwater. The results showed that in the tested water samples, the concentrations of Zn and Mn were the highest, and Ni was the lowest. Higher abundances of ARGs with great proportion of sulfonamides, chloramphenicols and tetracyclines resistance genes were found in mining-affected water when compared with those without mining activities. Additionally, there were positive correlations between heavy metals (especially Ni, Zn and Mn) and these ARGs. Linear regression analysis suggested that MGEs were positively correlated with ARGs. In addition, total phosphorus was correlated with ARGs (p < 0.05). The microbial community was different between the mining-affected water and the reference (p < 0.05). Proteobacteria, Bacteroidetes and Actinobacteria were dominant phyla in the surface water and groundwater. Network analysis showed that many ARGs were significantly associated with these dominant bacteria, which suggested they might be potential hosts for these ARGs. These findings provide a clear evidence that the mining activities in the study area had a significant impact on surface water and groundwater to different degrees.

Highly enhanced biodegradation of pharmaceutical and personal care products in a novel tidal flow constructed wetland with baffle and plants.

Research on decentralized wastewaters deserves special focus due to the potential abundance of emerging organic pollutants including pharmaceutical and personal care products (PPCPs), which might pose serious threats to the local water bodies and even to human health. Constructed wetland (CW) is a common decentralized wastewater treatment technology, with a certain ability to eliminate PPCPs. Nonetheless, PPCPs removal in common CWs is frequently challenging, besides, the removal mechanism remains elusive. Based on our previous study, tidal flow constructed wetlands (TFCWs) is effective in nitrogen removal. Here, 3 TFCWs with different modifications (baffle, plants, both baffle and plants) were constructed to treat raw domestic sewage and specifically to evaluate the removal efficiencies and mechanism of PPCPs. 24 PPCPs including 7 antibiotics, 8 steroid hormones and 9 biocides were detected in the level of 1.10 ± 0.29 ng/L-799 ± 10.6 ng/L in the influents. Consequently, we found that modification with both baffle and plants significantly influenced the removal of PPCPs. Moreover, the highest removal rates of biocides (97.1 ± 0.29%), steroid hormones (99.8 ± 0.02%), and antibiotics (90.2 ± 1.60%) were achieved via both baffles and plants in TFCWs. Based on the mass balance analysis, microbial degradation dominated the removal of PPCPs with a percentage higher than 85.7%, followed by substrate adsorption (5.22 × 10-2-14.3%) and plant uptake (1.66 × 10-3-0.44%). Further, 16S rDNA sequencing analysis revealed that the presence of baffle and plants improve the removal efficiency of PPCPs by means of enhancing microbial diversity and changing dominant microorganisms. Moreover, Thaumarchaeota was potentially the key microorganism in the phylum level for PPCPs elimination by TFCWs through LEfSe (linear discriminant analysis (LDA) effect size) analysis. These findings provide new insights into the removal of PPCPs in CWs.

Trace analysis of 28 antibiotics in plant tissues (root, stem, leaf and seed) by optimized QuEChERS pretreatment with UHPLC-MS/MS detection.

Phytoremediation has proven to be an effective in-situ treatment technique for antibiotic contamination. Due to the immature methods of extracting multi-antibiotics in different plant tissues, the antibiotic absorption and transportation mechanism in the phytoremediation process has yet to be resolved. Therefore, an improved Quick, Easy, Cheap, Effective, Rugged and Safe (QuEChERS) pretreatment with ultra-high-performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS) detection method for 28 antibiotics in different plant tissues (root, stem, leaf and seed) was developed in this study. The optimized method showed satisfactory performance with recoveries for most antibiotics ranging from 70% to 130% (except sulfadoxine with 138 ± 8.84% in root, sulfameter with 68.9 ± 1.87% and sulfadoxine with 141 ± 10.0% in seed). The limits of detection (LODs) of the target compounds in root, stem, leaf and seed were 0.04 ± 0.02 ~ 2.50 ± 1.14 ng/g, 0.05 ± 0.02 ~ 1.78 ± 0.42 ng/g, 0.06 ± 0.01 ~ 2.50 ± 0.14 ng/g and 0.13 ± 0.10 ~ 3.64 ± 0.74 ng/g, respectively. This developed method was successfully applied to the determination of antibiotics in different tissues of hydroponic wetland plants exposed to antibiotics-spiked water for one-month. Sixteen of 28 spiked antibiotics were detected in plant tissue samples. Overall, of these 16 antibiotics, all were detected in root samples (from < LOQ to 1478 ± 353 ng/g), eleven in stem samples (from < LOQ to 425 ± 47.0 ng/g), and nine in leaf samples (from < LOQ to 429 ± 84.5 ng/g). This developed analytical method provided a robust tool for the simultaneous screening and determination of antibiotics in different plant tissues.

[Profiles and Risk of Antibiotic Resistance Genes in Domestic Wells in the Maozhou River Basin].

Intensive use of antibiotics promotes the occurrence and development of antibiotic resistance. Antibiotic resistance genes (ARGs) enter water environments from human and animal sources. Groundwater serves as an important water supply, while the profiles and risk of ARGs in groundwater remain unknown. The abundance and profiles of ARGs in 11 domestic wells in the Maozhou River basin of Shenzhen City were analyzed by high-throughput qPCR. The results showed that a total of 141 ARGs and 8 mobile genetic elements (MGEs) were detected, of which the genes corresponding resistance to sulfonamides, multidrugs, and aminoglycosides were the most abundant. In each well, the number of detected ARGs and MGEs ranged from 48 to 89, with an average of 68. When normalized by the abundance of 16S rRNA genes, it was found that each bacterium carried at least one ARG in the groundwater of W7, W8, and W10, while in W11, each bacterium carried at least four ARGs. Clinically relevant ARGs that code for resistance to glycopeptide (blaSHV, blaTEM, blaCTX, and blaOXA-1), ß-lactams (vanB and vanC-03), or chloramphenicol (floR) were found in groundwater. In addition, the abundance of sulfonamides, multidrugs, aminoglycosides, ß-lactam, and chloramphenicol resistance genes were positively correlated with the abundance of MGEs (P<0.01), suggesting that MGEs may promote the spread of ARGs in groundwater.

Untreated swine wastes changed antibiotic resistance and microbial community in the soils and impacted abundances of antibiotic resistance genes in the vegetables.

Animal waste fertilization is a traditional agricultural practice, which may have adverse effects to soil ecosystem. However, the side-effects of animal waste fertilization on vegetables are less studied. Here we selected a swine farming village for investigation with a nearby village without swine farming as comparison. In the swine farming village, the farmers use untreated swine manure and wastewater as fertilizers for vegetable cultivation. In the reference village, the farmers mainly use commercial organic fertilizers. The objective of this study is to assess the impacts of untreated swine waste fertilization on both soils and vegetables in terms of antibiotics, antibiotic resistance genes (ARGs) and bacterial microbial communities. The results indicate that untreated swine waste fertilization caused both antibiotic and ARG contaminations and changed the microbial community compositions in the soils. Varieties of tetracyclines and related resistance genes were detected especially in swine wastewater treated soils. The soil quality was impacted with the relations to bacterial abundances and microbial geochemical functions. Proteobacteria and Bacteroidetes were prevalent and positively correlated to ARGs in soils, indicating they were potential antibiotic resistant bacteria. Antibiotics and ARGs were detected in vegetables of both villages. The abundances of ARGs were relatively higher in some vegetable samples of the swine farming village than the reference village. In addition, intracellular parasites Rickettsiales with positive correlation to ARGs were prevalent in some vegetables of swine farming village, indicating potential health risks through eating contaminated vegetables. The results of this study suggest that untreated swine wastes may cause adverse effects to not only agricultural soils but also associated vegetables.

Contamination profile of antibiotic resistance genes in ground water in comparison with surface water.

Dissemination of antibiotic resistance genes (ARGs) in the water environment has become an increasing concern. There have been many reports on ARGs in surface water, but little is known about ARGs in groundwater. In this study, we investigated the profiles and abundance of ARGs in groundwater in comparison with those in surface water of Maozhou River using high-throughput quantitative PCR (HT-qPCR). Totally 127 ARGs and 10 MGEs were detected by HT-qPCR, and among them the sulfonamides, multidrug and aminoglycosides resistance genes were the dominant ARG types. According to the results of HT-qPCR, 18 frequently detected ARGs conferring resistance to 6 classes of antibiotics and 3 MGEs were further quantified by qPCR in the wet season and dry season. The absolute abundance ranged from 1.23 × 105 to 8.89 × 106 copies/mL in wet season and from 8.50 × 102 to 2.65 × 106 copies/mL in the dry season, with sul1 and sul2 being the most abundant ARGs. The absolute abundance of ARGs and MGEs has no significant difference between the wet season and dry season while the diversity of ARGs in the dry season was higher than that in the wet season (p < 0.05). Totally 141 and 150 ARGs were detected in the water and sediments of Maozhou River, respectively. A total of 116 ARGs were shared among the groundwater, river water, and sediment, which accounted for 67.1% of all detected genes. Redundancy analysis further demonstrated that the environmental factors contributed 70.7% of the total ARG variations. The findings of large shared ARGs, abundant Total Coliforms and large wastewater burden in the groundwater provide a clear evidence that anthropogenic activities had a significant impact on groundwater.