High Abundance Of Antibiotic Resistance Genes Research Articles

Seasonal variation of antibiotic resistance genes in activated sludge of a full-scale municipal wastewater treatment plant: Contribution of activated sludge functional taxa and clinically relevant taxa

It has been demonstrated that antibiotic resistance genes (ARGs) exhibit seasonal variations in municipal wastewater treatment plants (MWTPs), but their relationship to bacterial phylogeny structure remains unclear. Using advanced metagenomic techniques and machine learning approach, the current study conducted a year-long investigation to explore the relationship between ARGs and the bacterial community of activated sludge in a full-scale MWTP in Beijing, where seasonal dynamics are remarkable. High abundance of ARGs, notably the clinically relevant high-risk ARGs, was observed in winter and spring, the cold season in Beijing. Seasonal patterns were also observed in the diversity of ARGs and the overall bacterial community. Machine learning-based random forest classification models were utilized to identify biomarkers for ARGs and bacterial genera as indicators of seasonal differences. Subsequent analysis of the relationship between ARGs and bacterial biomarkers was examined using random forest regression models. Results showed that the enrichment of potential pathogens such as Mycobacterium, Clostridium and Pseudomonas was high in winter and spring, strongly contributing to the abundance of high-risk ARGs (ermB, aac(6′)-I, tetM, blaTEM, and mefA) during cold season. Conversely, functional taxa associated with activated sludge, such as Thauera, displayed seasonal fluctuations and a preference for ARGs with minor clinical implications. Metagenomic binning further illustrated the contribution of Mycobacterium to ARG enrichment in cold season. Our findings highlight the collective impact of human-derived clinically relevant taxa and functional bacterial taxa in activated sludge on the seasonal dynamics of ARGs in MWTPs. Additionally, this study offers valuable insights into the safe disposal of the excess sludge from MWTPs.

Metagenomic exploration of Andaman region of the Indian Ocean

Ocean microbiome is crucial for global biogeochemical cycles and primary productivity. Despite numerous studies investigating the global ocean microbiomes, the microbiome composition of the Andaman region of the Indian Ocean remains largely unexplored. While this region harbors pristine biological diversity, the escalating anthropogenic activities along coastal habitats exert an influence on the microbial ecology and impact the aquatic ecosystems. We investigated the microbiome composition in the coastal waters of the Andaman Islands by 16S rRNA gene amplicon and metagenomic shotgun sequencing approaches and compared it with the Tara Oceans Consortium. In the coastal waters of the Andaman Islands, a significantly higher abundance and diversity of Synechococcus species was observed with a higher abundance of photosynthesis pigment-related genes to adapt to variable light conditions and nutrition. In contrast, Prochlorococcus species showed higher abundance in open ocean water samples of the Indian Ocean region, with a relatively limited functional diversity. A higher abundance of antibiotic-resistance genes was also noted in the coastal waters region. We also updated the ocean microbiome gene catalog with 93,172 unique genes from the Andaman coastal water microbiome. This study provides valuable insights into the Indian Ocean microbiome and supplements the global marine microbial ecosystem studies.

Ozone micron bubble pretreatment for antibiotic resistance genes reduction in hospital wastewater treatment

Ozone micron bubble (OMB) treatment offers a promising approach to effectively eliminate Antibiotic Resistance Genes (ARGs) from infectious medical wastewater and mitigate the threat of drug resistance transmission. This study evaluated the effectiveness of OMB treatment for reducing ARGs from infectious medical wastewater in laboratory and on-site pilot treatment setups. In part, the presence of antibiotic residues in a hospital wastewater treatment plant (WWTP) and the impact of hospital wastewater on the distribution of ARGs in a wastewater collection system were also investigated. The results of wastewater collection system survey revealed a high prevalence of ARGs in the system, particularly mcr-1, largely originating from medical wastewater discharges. Furthermore, analysis of antibiotic residues in the hospital wastewater treatment system showed significant accumulation, particularly of quinolone antibiotics, in the biomass of the biological oxidation tank, suggesting a potential risk of ARG proliferation within the system. Comparison of wastewater samples from domestic and hospital WWTPs revealed a relatively higher abundance of ARGs in the latter, with differences ranging from 2.2 to sixfold between corresponding locations in the treatment plants. Notably, the biological oxidation unit of both WWTPs exhibited a greater proportion of ARGs among all sampled points, indicating the potential proliferation of ARGs within the biomass of the treatment units. ARG degradation experiments showed that OMB treatment resulted in a significantly lower CT value (9.3 mg O3 L−1 min) compared to ozone coarse bubble treatment (102 mg O3 L−1 min) under identical test conditions. Moreover, the use of OMB on site significantly reduced the accumulation of ARGs in hospital wastewater, underscoring its potential as an effective solution for mitigating ARG spread.

Assessing the nonlinear association of environmental factors with antibiotic resistance genes (ARGs) in the Yangtze River Mouth, China

The emergence of antibacterial resistance (ABR) is an urgent and complex public health challenge worldwide. Antibiotic resistant genes (ARGs) are considered as a new pollutant by the WHO because of their wide distribution and emerging prevalence. The role of environmental factors in developing ARGs in bacterial populations is still poorly understood. Therefore, the relationship between environmental factors and bacteria should be explored to combat ABR and propose more tailored solutions in a specific region. Here, we collected and analyzed surface water samples from Yangtze Delta, China during 2021, and assessed the nonlinear association of environmental factors with ARGs through a sigmoid model. A high abundance of ARGs was detected. Amoxicillin, phosphorus (P), chromium (Cr), manganese (Mn), calcium (Ca), and strontium (Sr) were found to be strongly associated with ARGs and identified as potential key contributors to ARG detection. Our findings suggest that the suppression of ARGs may be achieved by decreasing the concentration of phosphorus in surface water. Additionally, Group 2A light metals (e.g., magnesium and calcium) may be candidates for the development of eco-friendly reagents for controlling antibiotic resistance in the future.

Transmission and retention of antibiotic resistance genes (ARGs) in chicken and sheep manure composting

Transmission of ARGs during composting with different feedstocks (i.e., sheep manure (SM), chicken manure (CM) and mixed manure (MM, SM:CM = 3:1 ratio) was studied by metagenomic sequencing. 53 subtypes of ARGs for 22 types of antibiotics were identified as commonly present in these compost mixes; among them, CM had higher abundance of ARGs, 1.69 times than that in SM, while the whole elimination rate of CM, MM and SM were 55.2%, 54.7% and 42.9%, respectively. More than 50 subtypes of ARGs (with 8.6%, 11.4% and 20.9% abundance in the initial stage in CM, MM and SM composting) were “diehard” ARGs, and their abundance grew significantly to 56.5%, 63.2% and 69.9% at the mature stage. These “diehard” ARGs were transferred from initial hosts of pathogenic and/or probiotic bacteria to final hosts of thermophilic bacteria, by horizontal gene transfer (HGT) via mobile gene elements (MGEs), and became rooted in composting products.

Plastic wastes and surface antibiotic resistance genes pollution in mangrove environments.

Mangroves are located at the intersection of land and sea and are also heavily affected by plastic wastes. Biofilms of plastic wastes in mangroves are reservoirs for antibiotic resistance genes (ARGs). In this study, plastic wastes and ARG pollution were investigated from three typical mangrove areas in Zhanjiang, South China. Transparent was the dominant colors of plastic wastes in three mangroves. Fragment and film shape accounted for 57.73-88.23% of plastic waste samples in mangroves. In addition, 39.50% of plastic wastes in protected area mangroves are PS. The metagenomic results shows that the 175 ARGs were found on plastic wastes of the three mangroves, the abundance accounting for 91.11% of the total ARGs. The abundance of Vibrio accounted for 2.31% of the total bacteria genera in aquaculture pond area mangrove. Correlation analysis shows that a microbe can carry multiple ARGs that may improve resistance to antibiotics. Microbes are the potential hosts of most ARGs, suggesting that ARGs can be transmitted by microbes. Because the mangroves are closely related to human activities and the high abundance of ARGs on plastic increases the ecological risks, people should improve plastic waste management and prevent the spread of ARGs by reducing plastic pollution.

The unique features of aerobic granule sludge contribute to simultaneous antibiotic removal and mitigation of antibiotic resistance genes enrichment

Antimicrobial resistance is now an increasingly serious threat to public health. In this study, the distribution and enrichment of antibiotic resistance genes (ARGs) in aerobic granular sludges (AGS) during sulfamethoxazole (SMX) biodegradation were examined. The removal percentage of SMX kept higher than 99 % at the loading of 750 μg/L after the formation of AGS. Through quantitative polymerase chain reaction (qPCR) and metagenomics, we showed that higher abundance of ARGs were in flocs (143 ARG subtypes, 3.08 ± 0.99 × 107 copies per ngDNA) and the outer AGS (111 ARG subtypes, 3.50 ± 0.25 × 107 copies per ngDNA) than that in the inner AGS (102 ARG subtypes, 8.10 ± 0.79 × 106 copies per ngDNA). Two mechanisms may explain the simultaneous antibiotic removal and mitigation of ARG enrichment by AGS. Firstly, dense aggregated structure of AGS and the abundant extracellular polymeric substances (EPS) play an important role in mitigating ARGs enrichment by preventing antibiotic transfer into the inner space of AGS. The deterministic mechanisms drive AGS bacterial community assemble (βNTI > 2) in a direction toward accumulating EPS producers. Secondly, the higher abundance of antibiotic co-metabolic genes in AGS than flocs revealed that functional bacteria in the outer AGS may decrease antibiotic pressures for the inner bacteria by co-metabolic degradation of antibiotics through nitrification and denitrification. This work discovered that AGS harbor ARG-independent resistance and degradation of antibiotics, and provided a new perspective for biological wastewater treatment.

Deciphering of microbial community and antibiotic resistance genes to the selection pressures of antibiotic concentration in aerobic activated sludge reactors

A high antibiotic concentration promotes the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs) in the wastewater while reducing the ARG abundance due to the production of resistance. However, the two-way effects of antibiotic concentration on the abundance of ARGs with different antibiotics are still lacking. Herein, the fate of ARGs and the corresponding composition of the microbial community (i.e., resistome and potential host bacteria for HGT) were investigated by adding two typical antibiotics (i.e., sulfadiazine, SDZ, and tetracycline hydrochloride, TC-HCl) under different concentrations. Results indicated that the initial antibiotic concentration for two antibiotics had different effects on the abundance of the corresponding ARGs (i.e., tet or sul); however, it was affected by the final antibiotic concentration, and a higher final antibiotic concentration would have a higher ARG abundance for both two antibiotic treatments. A higher initial antibiotic concentration would decrease the abundance but improve the diversity of the microbial community, resulting in the different biodegradation abilities of the corresponding antibiotic and subsequently affecting the resistome of microbial community, resulting in altering the abundance of ARGs (i.e., tet or sul). On the other hand, a higher final antibiotic concentration had a higher abundance of potential host bacteria, resulting in a higher abundance of ARGs (i.e., tet or sul). This study provides new insights into the mechanisms underlying the alteration of ARG abundance to the selection pressures of antibiotic concentration in aerobic activated sludge reactors.

High occurrence of antibiotic resistance genes in intensive aquaculture of hybrid snakehead fish

The increasing abundance of antibiotic resistance genes (ARGs), which are regarded as new pollutants, has raised public health concern. The use of antibiotics in aquaculture has promoted the evolution and spread of ARGs. The occurrence and abundance of ARGs in aquaculture has attracted extensive attention. However, the distribution and transmission of ARGs in aquaculture require further study. This study analyzed water and sediment from intensive culture of hybrid snakehead fish farm in Zhongshan, South China. Twenty-two types of ARGs were detected in all environmental samples. The relative abundance of sulfonamide resistance genes (sul1 and sul2) was the highest, ranging from 3.37×10-2 to 8.53×100 copies/16S rRNA gene. High occurrence of quinolones, phenicols, tetracycline resistance genes, and class 1 integrase gene (intI1) was also observed in pond water samples. This implies that pond water is one of the main reservoirs and origins of ARGs in the aquatic environment. Proteobacteria was the most abundant phylum in all the environmental samples, and its relative abundance ranged from 24.05% to 41.84%. Network and canonical correspondence analyses showed that a high abundance of ARGs (int1, sul1, sul2, qacEΔ1, aac6, and oqxA) was positively correlated with Proteobacteria, Cyanobacteria, and Bacteroidetes, and the abundance of Proteobacteria, Actinobacteria, and Patescibacteria was positively correlated with environmental factors (sulfide, nitrite nitrogen, pH, free chlorine, and ammonia nitrogen). These findings demonstrate the prevalence and persistence of ARGs in intensive fish farming in southern China. This suggests that ARG levels and microbiological community composition in aquaculture should be conventionally determined to assess potential risks to public health.

Tracking antibiotic resistance genes in microplastic-contaminated soil

Agricultural soils and microplastics (MPs) are hotspots for antibiotic resistance genes (ARGs). Plastic mulch is the most important source of MPs in agricultural soil. ARGs, mobile genetic elements (MGEs), and their host profiles in long-term mulch MP-exposed soils remain unclear. In the present study, metagenomics was used to investigate the distribution patterns of ARGs and MGEs in eight Chinese provinces with a long history of plastic mulch use. A total of 204 subtypes of ARGs and thousands of MGEs (14 integrons, 28 insertions, and 2993 plasmids) were identified. A similar diversity of ARGs was found among MPs film-contaminated sites. The types of ARGs with a high abundance were more concentrated, and multidrug resistance genes were the dominant ARGs. Soils from regions with a longer history of plastic film use (such as Xinjiang province) had a higher abundance of ARGs and MGEs. The distribution of ARGs and MGEs exhibited a modular network distribution pattern. A total of 27 ARG subtypes and 29 MGEs showed co-occurrence network relationships. More than 10 common hosts of ARGs and MGEs, such as Pseudomonas, were found, and their abundances were highest in three provinces, including Xinjiang. This study may help elucidate the impact mechanism of long-term MP residues on the occurrence and spread of ARGs in soil.

Distribution and migration of antibiotic resistance genes, as well as their correlation with microbial communities in swine farm and its surrounding environments

The prevalence and correlation of antibiotic resistance genes (ARGs) in pig farm wastewater treatment plants (WWTPs) and surrounding environment were investigated using metagenomics and real time quantitative PCR (q-PCR). The hosts of ARGs were also studied in this study. The abundance of ARGs decreased significantly in the anoxic/oxic (A/O) process and disinfection tank of WWTPs. New ARGs emerged in wastewater that passed though the anaerobic reactor. The abundances of ARGs in the soils and water near pig farm were 10- and 35-fold higher than those in the control, respectively. The abundance of ARGs in wells near pig farm were an order of magnitude higher than that in the control. Similarly, a high abundance of ARGs was detected in swine manure. After composting, most of the ARGs were eliminated, but sul1 increased 10.5-fold. A high-throughput analysis revealed that the pig farm altered the microbial community structure in the surrounding environment, with 52% and 37% of the operational taxonomic units (OTUs) endemic to the soil and water samples near pig farm in comparison with these data in the control, respectively. The phyla Proteobacteria, Choroflexi, and Actinobacteriota dominated the water and soil samples. In addition, three pathogenic genera were found in the surrounding soil and water samples. A metagenomic analysis identified 14 types of ARGs (>1%), with the highest proportion of multidrug ARGs at 47%. A total of 28 subtypes of ARGs were detected (>1%), with macB the most prevalent. The correlation analysis revealed that several key phyla, including Proteobacteria, Actinobacteria and Acidobacteria, were the main potential hosts and posed a positive correlation with the ARGs. Efflux pumps (60–66%) were the primary resistance mechanism, and each resistance mechanism was distributed in similar proportions in the microbial community.

Diversity and abundance of antibiotic resistance genes and their relationship with nutrients and land use of the inflow rivers of Taihu Lake.

Taihu Lake is the third largest freshwater lake in China and an important source for drinking water, flood protection, aquaculture, agriculture, and other activities. This lake is connected to many principal and small rivers with inflow from west and outflow on the eastern side of the lake and these inflow rivers are believed to significantly contribute to the water pollution of the lake. This study was aimed at assessing the diversity and abundance of antibiotic resistance genes (ARGs) and mobile genetic elements (MGEs), and their relationship with water quality parameters and land use patterns. Water samples were collected from 10 major inflow rivers and the source water protection area of the Taihu Lake in spring and summer 2019. High-throughput profiling was used to detect and quantify 384 ARGs and MGEs and in addition, 11 water quality parameters were analyzed. The results showed that the number of ARGs/MGEs detected in each inflow river ranged from 105 to 185 in spring and 107 to 180 in summer. The aminoglycoside resistance genes were the most dominant types ARGs detected followed by beta-lactam resistance, multidrug resistance, macrolide-lincosamide-streptogramin B (MLSB) resistance genes, which contributed to 65% of the ARGs. The water quality parameters showed significant correlation with absolute abundance of ARGs. Furthermore, significant correlation between ARGs and MGEs were also observed which demonstrates potential gene transfer among organisms through horizontal gene transfer via MGEs. ARGs showed strong positive correlation with cultivated and industrial lands whereas, negative correlation was observed with river, lake, forest, land for green buffer, and land for port and harbor. The overall results indicate that the inflow rivers of Taihu Lake are polluted by various sources including multiple nutrients and high abundance of ARGs, which needs attention for better management of the inflow rivers of this lake.

Shotgun metagenomic analysis reveals the prevalence of antibiotic resistance genes and mobile genetic elements in full scale hospital wastewater treatment plants

Wastewater treatment plants are considered as hotspots of emerging antimicrobial genes and mobile genetic elements. We used a shotgun metagenomic approach to examine the wide-spectrum profiles of ARGs (antibiotic resistance genes) and MGEs (mobile genetic elements) in activated sludge samples from two different hospital trains at the wastewater treatment plants (WWTPs) in Daegu, South Korea. The influent activated sludge and effluent of two trains (six samples in total) at WWTPs receiving domestic sewage wastewater (SWW) and hospital wastewater (HWW) samples collected at multiple periods were subjected to high throughput 16S rRNA metagenome sequencing for microbial community diversity. Cloacibacterium caeni and Lewinella nigricans were predominant in SWW effluents, while Bacillus subtilis and Staphylococcus epidermidis were predominant in HWW effluents based on the Miseq platform. Totally, 20,011 reads and 28,545 metagenomic sequence reads were assigned to 25 known ARG types in the SWW2 and HWW5 samples, respectively. The higher abundance of ARGs, including multidrug resistance (>53%, MDR), macrolide-lincosamide-streptogramin (>9%, MLS), beta-lactam (>3.3%), bacitracin (>4.4%), and tetracycline (>3.4%), confirmed the use of these antibiotics in human medicine. In total, 190 subtypes belonging to 23 antibiotic classes were detected in both SWW2 and HWW5 samples. RpoB2, MacB, and multidrug (MDR) ABC transporter shared the maximum matched genes in both activated sludge samples. The high abundance of MGEs, such as a gene transfer agent (GTA) (four times higher), transposable elements (1.6 times higher), plasmid related functions (3.8 times higher), and phages (two times higher) in HWW5 than in SWW2, revealed a risk of horizontal gene transfer in HWW. Domestic wastewater from hospital patients also influenced the abundance of ARGs and MGEs in the activated sludge process.

Ozone pretreatment of wastewater containing aromatics reduces antibiotic resistance genes in bioreactors: The example of p-aminophenol

Aromatic matters are widely present in wastewater, especially industrial wastewater, and may lead to a high abundance of antibiotic resistance genes (ARGs) in wastewater treatment bioreactors and stimulate horizontal transfers of ARGs. Here, we investigated a practical approach that applying ozone pretreatment to mitigate ARGs in bioreactors treating wastewater containing a typical aromatic pollutant, p-aminophenol (PAP). The results showed that ozone pretreatment could effectively reduce the aromaticity of wastewater, and the relative abundance of ARGs in the bioreactor fed with ozone treated wastewater decreased by over 70% compared to the control reactor. Multidrug, quinolone, mupirocin, polymyxin, aminoglycoside, glycopeptide, beta-lactam, and trimethoprim resistance genes were all reduced in the bioreactors receiving wastewater pretreated by ozone. Metagenomic analysis suggested that the reduction of ARGs could be attributed to the co-occurrence of ARGs and aromatic degradation genes in bacteria. Furthermore, we expanded our analysis to investigate 71 metagenomes from different environments, and the results indicated that the impact of aromatics on ARG abundance widely occurs in various ecosystems and confirmed that high levels of aromatics could lead to high abundance of ARGs. Taken together, our work confirmed that the aromatics played critical roles in selecting ARGs and proposed a feasible approach to reduce ARGs in wastewater treatment bioreactors.

Human Milk Microbiota: Transferring the Antibiotic Resistome to Infants.

Commensal bacterial population is believed to be a reservoir for antibiotic resistance genes (ARGs). The infant gut microbiota has relatively higher abundance of ARGs than the adults. These genes can get transferred from commensals to pathogens by horizontal gene transfer, which magnifies the spectrum of antibiotic resistance in the environment. The presence of ARGs in neo-nates and infants, with no prior antibiotic exposure, questions their origin in the naïve commensal population. Breast milk microbiota that is responsible for the initial seeding of infant gut microbiota has also been found to harbour a vast array of ARGs. This review discusses the recent findings that indicate the potential of breast milk microbiota to act as a vehicle for transmission of ARGs to infants.

Distribution characteristics of antibiotic resistant bacteria and genes in fresh and composted manures of livestock farms

Livestock manure is a major reservoir of antibiotic resistant bacteria (ARB) and antibiotic resistance genes (ARGs). This study investigated the distribution characteristics of ARB, ARGs in fresh and composted manures of traditional breading industry in rural areas in China. Samples collected were naturally piled without professional composting, and will be applied to farmland. The real-time quantitative polymerase chain reaction (qPCR) results showed the presence of ten target ARGs and two mobile genetic elements (MGEs) in the tested manure samples. The relative abundance of tetracycline and sulfonamide resistance genes (TRGs and SRGs) was generally higher than that of macrolide resistance genes (MRGs), followed by quinolone resistance genes (QRGs). There were significant positive correlations between the abundance of sul1, sul2, tetW and MGEs (intl1, intl2). In addition, the distribution of target ARGs was associated with the residual concentrations of doxycycline (DOX), sulfamethazine (SM2), enrofloxacin (ENR) and tylosin (TYL). Overall, a total of 24 bacterial genera were identified. The resistance rates of ARB were 17.79%–83.70% for SM2, followed 0.40%–63.77% for TYL, 0.36%–43.90% for DOX and 0.00%–13.36% for ENR, which showed a significant dose-effect. This study also demonstrated that the abundance of clinically relevant ARB and ARGs in chicken, swine and cow fresh manures significantly greater than that in composted manures, and chicken and swine manures had higher proportion of ARB and higher abundance of ARGs than that in cow manures.

Pharmaceutical exposure changed antibiotic resistance genes and bacterial communities in soil-surface- and overhead-irrigated greenhouse lettuce

New classes of emerging contaminants such as pharmaceuticals, antibiotic resistant bacteria (ARB), and antibiotic resistance genes (ARGs) have received increasing attention due to rapid increases of their abundance in agroecosystems. As food consumption is a direct exposure pathway of pharmaceuticals, ARB, and ARGs to humans, it is important to understand changes of bacterial communities and ARG profiles in food crops produced with contaminated soils and waters. This study examined the level and type of ARGs and bacterial community composition in soil, and lettuce shoots and roots under soil-surface or overhead irrigation with pharmaceuticals-contaminated water, using high throughput qPCR and 16S rRNA amplicon sequencing techniques, respectively. In total 52 ARG subtypes were detected in the soil, lettuce shoot and root samples, with mobile genetic elements (MGEs), and macrolide-lincosamide-streptogramin B (MLSB) and multidrug resistance (MDR) genes as dominant types. The overall abundance and diversity of ARGs and bacteria associated with lettuce shoots under soil-surface irrigation were lower than those under overhead irrigation, indicating soil-surface irrigation may have lower risks of producing food crops with high abundance of ARGs. ARG profiles and bacterial communities were sensitive to pharmaceutical exposure, but no consistent patterns of changes were observed. MGE intl1 was consistently more abundant with pharmaceutical exposure than in the absence of pharmaceuticals. Pharmaceutical exposure enriched Proteobacteria (specifically Methylophilaceae) and decreased bacterial alpha diversity. Finally, there were significant interplays among bacteria community, antibiotic concentrations, and ARG abundance possibly involving hotspots including Sphingomonadaceae, Pirellulaceae, and Chitinophagaceae, MGEs (intl1 and tnpA_1) and MDR genes (mexF and oprJ).

Dynamic transport of antibiotics and antibiotic resistance genes under different treatment processes in a typical pharmaceutical wastewater treatment plant.

The propagation of antibiotic resistance is a challenge for human health worldwide, which has drawn much attention on the reduction of the resistance genes. To understand their occurrence during different treatment processes, in this study, four classes of antibiotics (tetracyclines, sulfonamides, quinolones, and macrolides), eight antibiotic resistance genes (ARGs) (tetB, tetW, sul1, sul2, gyrA, qepA, ermB, and ermF), and two mobile elements (int1 and int2) were investigated in a typical pharmaceutical plant. The total concentrations of antibiotics were detected in the range of 2.6×102 to 2.5×103ng/L in the treatment processes, and the high abundance of ARGs was detected in the biological treatment unit. The dynamic trend analysis showed that antibiotics were partially removed in the anaerobic/aerobic processes, where ARGs were proliferated. The abundance of tetB and gyrA genes was positively correlated with pH and EC (p< 0.05), and the tetW, sul1 and sul2 genes were significantly correlated with TOC, TN, and DO (p< 0.05), indicating the influence of physicochemical properties of the solution on the levels of ARG subtypes. The phylogenetic analysis showed that the tetW clones had high homology with some pathogenic microorganisms, such as Klebsiella pneumonia and Neisseria meningitides, which would threaten human health. Results indicated that the horizontal transfer acted as a major driver in the ARGs evolution.

Cultivation-dependent and high-throughput sequencing approaches studying the co-occurrence of antibiotic resistance genes in municipal sewage system.

During the past years, antibiotic-resistant bacteria (ARB) leading for the spreading of antibiotic resistance genes (ARGs) became a global problem, especially multidrug-resistant (MDR) bacteria are considered the prime culprit of antibiotic resistance. However, the correlation between the antibiotic-resistant phenotype and the ARG profiles remains poorly understood. In the present study, metagenomic functional screening and metagenomic analysis of coliforms were combined to explore the phenotype and genotype of the ARBs from municipal sewage. Our results showed that the ARG co-occurrence was widespread in the municipal sewage. The present study also highlighted the high abundance of ARGs from antibiotic resistance coliforms especially the MDR coliforms with ARG level of 33.8±4.2 copies per cell. The ARG profiles and the antibiotic resistance phenotypes of the isolated antibiotic resistant coliforms were also correlated and indicated that the resistance to the related antibiotic (ampicillin, kanamycin, erythromycin, chloramphenicol, and tetracycline) was mostly contributed by the ARGs belonging to the subtypes of β-lactamase, aminoglycoside 3-phosphotransferase, phosphotransferase type 2, chloramphenicol acetyltransferase, tetA, etc.

Feasibility of lettuce cultivation in sophoroliplid-enhanced washed soil originally polluted with Cd, antibiotics, and antibiotic-resistant genes

Vegetable cultivation in soils polluted with heavy metals, antibiotics and a high abundance of antibiotic-resistance genes (ARGs) can seriously threaten human health through the food chain. Therefore, novel techniques that not only remediate soil, but also ensure food security are urgently required. In the present study, two successive washings with 20gL−1 of sophoroliplid solution plus ultrasonication (35kHz) were effective in extracting 71.2% Cd, 88.2% tetracycline, 96.6% sulfadiazine, and 100% roxithromycin. Simultaneously, relative abundance of ARGs (tetM, tetX, sulI, and sulII) was decreased to 10−7–10−8 (ARG copies/16S copies). Further, lettuce cultivation in the 2nd washed soil showed significant improvement in vegetable growth indices (fresh/dry weight, root surface area, chlorophyll content and soluble protein content) and a decrease in isolate counts for antibiotic-resistant bacterial endophytes and ARG abundance in lettuce tissues. This combined cleanup strategy provides an environmentally friendly technology for ensuring vegetable security in washed soils.