Abundant Resistance Genes Research Articles

Exploring Urban Coastal Areas: Investigating the Urban Coastal Areas as a Reservoirs of Antibiotic Resistance Genes★

Antibiotic resistance genes (ARGs) have long served as adaptive defensive mechanisms among bacteria, enabling their survival and propagation in challenging environments. The consequences of inefficient wastewater treatment have culminated the emergence of untreatable and lethal extensively drug-resistant. To understand the relationship between wastewater effluent and marine ecosystems, we conducted a study to monitor the diversity and prevalence of common ARGs in Hong Kong's urban coastal areas at different seasons.Our findings revealed that sul 1 was the most abundant resistance gene, with an average relative abundance of 4.45×10-2 per 16s rRNA gene copy. Moreover, temperature, dissolved oxygen, and salinity were key factors influencing seasonal variations in total ARGs abundance. The influence of environmental factors varied based on ARGs' association with Intl1, with Intl1-associated ARGs strongly correlating with temperature and dissolved oxygen. Notably, despite their abundance, sul1 and mphA exhibited similar correlations with both Intl1 and key environmental factors, suggesting these ARGs share a common dissemination mechanism. Moreover, the robust association between resistance genes and mobile genetic elements (MGE) could potentially act as a valuable indicator for assessing the efficacy of removing ARGs in wastewater treatment methods when operating under carefully optimized environmental parameters.

Overexpression of Wild Soybean Expansin Gene GsEXLB14 Enhanced the Tolerance of Transgenic Soybean Hairy Roots to Salt and Drought Stresses.

As a type of cell-wall-relaxing protein that is widely present in plants, expansins have been shown to actively participate in the regulation of plant growth and responses to environmental stress. Wild soybeans have long existed in the wild environment and possess abundant resistance gene resources, which hold significant value for the improvement of cultivated soybean germplasm. In our previous study, we found that the wild soybean expansin gene GsEXLB14 is specifically transcribed in roots, and its transcription level significantly increases under salt and drought stress. To further identify the function of GsEXLB14, in this study, we cloned the CDS sequence of this gene. The transcription pattern of GsEXLB14 in the roots of wild soybean under salt and drought stress was analyzed by qRT-PCR. Using an Agrobacterium rhizogenes-mediated genetic transformation, we obtained soybean hairy roots overexpressing GsEXLB14. Under 150 mM NaCl- and 100 mM mannitol-simulated drought stress, the relative growth values of the number, length, and weight of transgenic soybean hairy roots were significantly higher than those of the control group. We obtained the transcriptomes of transgenic and wild-type soybean hairy roots under normal growth conditions and under salt and drought stress through RNA sequencing. A transcriptomic analysis showed that the transcription of genes encoding expansins (EXPB family), peroxidase, H+-transporting ATPase, and other genes was significantly upregulated in transgenic hairy roots under salt stress. Under drought stress, the transcription of expansin (EXPB/LB family) genes increased in transgenic hairy roots. In addition, the transcription of genes encoding peroxidases, calcium/calmodulin-dependent protein kinases, and dehydration-responsive proteins increased significantly. The results of qRT-PCR also confirmed that the transcription pattern of the above genes was consistent with the transcriptome. The differences in the transcript levels of the above genes may be the potential reason for the strong tolerance of soybean hairy roots overexpressing the GsEXLB14 gene under salt and drought stress. In conclusion, the expansin GsEXLB14 can be used as a valuable candidate gene for the molecular breeding of soybeans.

Evaluation of the Rhizosphere Resistome of Cultivated Soils Polluted with Antibiotics from Reclaimed Wastewater

The use of reclaimed wastewater to irrigate crops is a valuable option due to water scarcity. However, the presence of antibiotics residues that are not removed in the tertiary treatments reaches crop soils when irrigated and it poses a serious concern for human health. Crops rhizosphere is considered a hotspot of antibiotic resistant genes (ARG) being in addition a link to plant phyllosphere and human microbiome. Understanding the structure of the soil microbiota is crucial before applying any bioremediation or biostimulation strategy. The aim of this work was firstly to confirm the presence of antibiotics residues in soil and fruits in two greenhouses at the south of Spain irrigated with reclaimed water. Secondly, to characterize the rhizosphere microbiome of three crops (Capsicum annuum, Cucumis melo and Solanum melongena) cultured in those greenhouses. Finally, a predictive functional analysis was done using PICRUSt2 to figure out the rhizosphere resistome. The presence of residues of antibiotics was confirmed both soil and fruits. Antibiotics absorbed by plants correlated with those on soil. The most abundant resistance gene was the multidrug in all the three crops tested. Compatibility of basidiomycete fungi (i.e., Pleurotus) with soil bacteria could be indicative of their possible use for restoration of agricultural soils polluted with antibiotics.

Effects of different concentrations of biological maturity agents on nitrogen and microbial diversity of Auricularia heimuer residue compost

This study investigated the effects of different concentrations of biological maturity agents on the composting process of Auricularia heimuer residue by adding them to the composting process. By measuring the changes in physical and chemical indicators and microbial diversity during composting, the results showed that the addition of biological maturity agents had a certain promoting effect on compost temperature, humidity, pH, seed germination index, and vitality index. Appropriate composting days can promote the accumulation of ammonium nitrogen. The carbon content of humin and E4/E6 of treatments A, B, and E were significantly higher than those of the initial treatment. D0.CK treatment had the most types of resistance genes and the most abundant resistance genes. As composting progresses, the abundance of 13 resistance genes decreased. Adding high concentrations of biological maturity agents can activate the defense mechanism during the composting process, greatly ensuring the safety of fungi residue as a fertilizer.

Previous Antibiotic Exposure Reshapes the Population Structure of Infecting Uropathogenic Escherichia coli Strains by Selecting for Antibiotic Resistance over Urovirulence.

Antibiotic therapy is the standard of care for urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC). However, previous antibiotic therapy may impart a selective pressure that influences the population structure and pathogenic potential of infecting UPEC strains. Here, we conducted a 3-year study using whole-genome-sequencing analysis and retrospective medical record review to characterize how antibiotic exposure influenced the phenotypic antibiotic resistance, acquired resistome, virulome, and population structure of 88 UTI-causing E. coli strains from dogs. A majority of UTI-associated E. coli strains were from phylogroup B2 and clustered within sequence type 372. Previous antibiotic exposure was associated with a population shift toward UPEC from phylogroups other than the typical urovirulent phylogroup B2. The specific virulence profiles within the accessory virulome that were associated with antibiotic use were elicited by the effect of antibiotics on UPEC phylogenetic structure. Among phylogroup B2, antibiotic exposure increased the quantity of genes within the resistome and the odds of developing reduced susceptibility to at least one antibiotic. Non-B2 UPEC strains harbored a more diverse and greater resistome that conferred reduced susceptibility to multiple antibiotic classes following antibiotic exposure. Collectively, these data suggest that previous antibiotic exposure establishes an environment that provides a selective edge to non-B2 UPEC strains through their diverse and abundant antibiotic resistance genes, despite their lack of urovirulence genes. Our findings highlight the necessity for judicious use of antibiotics as we uncover another mechanism by which antibiotic exposure and resistance can influence the dynamics of bacterial infectious disease. IMPORTANCE Urinary tract infections (UTIs) are one of the most common infections of dogs and humans. While antibiotic therapy is the standard of care for UTIs and other infections, antibiotic exposure may influence the pathogenic profile of subsequent infections. We used whole-genome sequencing and retrospective medical record review to characterize the effect of systemic antibiotic therapy on the resistance, virulence, and population structure of 88 UTI-causing UPEC strains isolated from dogs. Our results indicate that antibiotic exposure alters the population structure of infecting UPEC strains, providing a selective edge for non-B2 phylogroups that harbor diverse and abundant resistance gene catalogues but fewer urovirulence genes. These findings highlight how antibiotic resistance can influence pathogen infection dynamics and have clinical implications for the judicious use of antibiotics for bacterial infections.

Emergence of Extensively Drug-Resistant ST170 Citrobacter portucalensis with Plasmids pK218-KPC, pK218-NDM, and pK218-SHV from a Tertiary Hospital, China.

ABSTRACTThe objective of this study is to characterize the molecular mechanism of a clinical carbapenem-resistant Citrobacter portucalensis strain K218, which coproduces KPC and NDM carbapenemases. K218 was isolated from a patient's blood sample in a Chinese tertiary hospital. Carbapenemases were detected by the immunocolloidal gold technique. The MIC values were determined by VITEK2. Whole-genome sequencing was performed on K218 and sequence data were analyzed using phylogenetics and extensive genomic comparison. This study reveals that K218 contains a single 5.08 Mb chromosome (51.8% GC content) and four plasmids, pK218-KPC (106 Kb), pK218-NDM (111 Kb), pK218-SHV (191 Kb), and pK218-NR (5 Kb). Twenty-nine types of antibiotic resistance genes were carried on K218, including blaKPC-2 harbored on pK218-KPC and blaNDM-1 harbored on pK218-NDM. Detailed comparison of related plasmids of pK218-KPC, pK218-NDM, and pK218-SHV showed that they shared similar conserved backbone regions, respectively. Comprehensive annotation revealed large accessory modules were recombined on the genome of K218. Further analysis speculated that mobile genetic elements bearing abundant resistance genes facilitated the formation of these accessory modules. In conclusion, this study provides an in-depth understanding of the genomic characterization of K218, an extensively drug-resistant C. portucalensis strain coproducing NDM and KPC carbapenemase. To the best of our knowledge, this is the first report of C. portucalensis strain coharboring blaKPC-2 and blaNDM-1 from the clinical setting.IMPORTANCE This is the first report of extensively drug-resistant C. portucalensis harboring both blaKPC-2 and blaNDM-1. This study will not only extend the understanding of the structural dissection of plasmids and chromosomes carried in C. portucalensis, but also expand knowledge of the genetic environment of the blaKPC-2 and blaNDM-1 genes. blaKPC-2 and blaNDM-1 genes have been suggested to facilitate the propagation and persistence of their host bacteria under different antimicrobial selection pressures. Large accessory regions carrying blaKPC-2 and blaNDM-1 genes have become hot spots for transposition and integration, and their structural variation and evolution should receive attention. The multidrug-resistant plasmids pK218-KPC, pK218-NDM, and pK218-SHV with several multidrug resistance regions and the chromosome cK218 with two novel transposons Tn7410 and Tn7411 contribute to the formation of extensively drug-resistant C. portucalensis.

A Novel Wheat-Rye 2R (2D) Disomic Substitution Line Pyramids Two Types of Resistance to Powdery Mildew.

Powdery mildew, caused by Blumeria graminis f. sp. tritici, is a devastating disease of wheat that seriously affects yield and quality worldwide. Because of the extensive growth of wheat cultivars with homogeneous genetic background, exploring novel resistant resources from wheat relatives has become important for increasing the genetic diversity of wheat. Rye (Secale cereale) is a wheat relative possessing abundant resistance genes because of its high variation. Wheat line AL69, resistant to powdery mildew, was developed by crossing, backcrossing, and self-pollination for multiple generations between hexaploid triticale Zhongsi 237 and common wheat cultivar Zimai 17. Through genomic in situ hybridization (GISH) and multicolor fluorescence in situ hybridization (FISH), nondenaturing FISH, multicolor GISH, and selection with specific molecular markers, AL69 was determined to be a wheat-rye 2R (2D) disomic substitution line. Testing with different B. graminis f. sp. tritici isolates and genetic analysis showed that the all-stage resistance (also called seedling resistance) of AL69 was conferred by the cataloged powdery mildew resistance gene Pm4b derived from Zimai 17, and its adult-plant resistance was derived from the alien chromosome 2R of Zhongsi 237, which was found to be different from the previously reported rye-derived Pm genes, including Pm7 on 2RL. In addition, AL69 showed improved spike number per plant, spike length, fertile spikelet number per spike, kernel number per spike, and grain yield per plant compared with its wheat parent Zimai 17. An elite line S251 combining powdery mildew resistance with excellent agronomic performance was selected from the progenies of AL69 and wheat cultivar Jimai 22. Therefore, AL69 has two types of resistance genes to powdery mildew and improved agronomic traits through pyramiding and thus can be used as a promising genetic stock for wheat breeding.

Genome-Wide Identification of Expansin Genes in Wild Soybean (Glycine soja) and Functional Characterization of Expansin B1 (GsEXPB1) in Soybean Hair Root

Wild soybean, the progenitor and close relative of cultivated soybean, has an excellent environmental adaptation ability and abundant resistance genes. Expansins, as a class of cell wall relaxation proteins, have important functions in regulating plant growth and stress resistance. In the present study, we identified a total of 75 members of the expansin family on the basis of recent genomic data published for wild soybean. The predicted results of promoter elements structure showed that wild soybean expansin may be associated with plant hormones, stress responses, and growth. Basal transcriptome data of vegetative organs suggest that the transcription of expansin members has some organ specificity. Meanwhile, the transcripts of some members had strong responses to salt, low temperature and drought stress. We screened and obtained an expansin gene, GsEXPB1, which is transcribed specifically in roots and actively responds to salt stress. The results of A. tumefaciens transient transfection showed that this protein was localized in the cell wall of onion epidermal cells. We initially analyzed the function of GsEXPB1 by a soybean hairy root transformation assay and found that overexpression of GsEXPB1 significantly increased the number of hairy roots, root length, root weight, and the tolerance to salt stress. This research provides a foundation for subsequent studies of expansins in wild soybean.

Study of the relationship between antibiotic resistance markers and virulence markers in NDM-positive Klebsiella pneumoniae strains circulating in various waters and human loci

Introduction. The propagation of multi-resistance to antibiotics among hospital isolates of Klebsiella pneumoniae (K. pneumoniae) is a subject of growing concern worldwide. At present, growing data of association between resistance and hypervirulence in clinical isolates of K. pneumoniae emerges. However, the occurrence of these pathogens in the environment remains an open question. The aim of this study was to evaluate and compare antibiotic resistance determinants occurrence in Klebsiella pneumoniae isolates from water sources (environmental and sewage), human sources (practically healthy people and patients with inflaammatory bowel disease (IBD), and extraintestinal infections (ExII)). Materials and methods. The PCR assay of carbapenemase genes IMP, NDM, VIM, KPC, OXA-48 was performed with the commercial “Amplisense” kits according to the manufacturer's instructions. The assay was used to evaluate the occurrence of antibiotic-resistance genes in 223 isolates of Klebsiella pneumoniae from various sources: 42 isolates from sewage, 19 isolates from surface water sources, 30 isolates from biological material (blood, urine, surgical wounds, bronchoalveolar lavage) of patients with extraintestinal infections (ExII), 69 isolates from patients with inflammatory bowel diseases (IBD), and 63 isolates from faeces of practically healthy people. Results. The ExII group revealed various antibiotic resistance genes. The most prevalent gene was OXA (30% had this gene only, other 26,6% had also KPC or NDM). NDM as the only resistance gene was observed in 23,3% of ExII isolates. KPC gene was observed in 3,3% of ExII group. Two isolates from IBD group contained NDM gene along with VIM gene. Only NDM gene was found in all the other groups of Klebsiella pneumoniae isolates (13-28% isolates in every group, no statistical difference). NDM was shown to be associated with virulence genes iutA and rmpA that are responsible for iron consumption and hypermucoid phenotype. Conclusion. The most abundant resistance genes in the studied Klebsiella pneumoniae isolates were NDM (13.5%) and OXA (8%). At the same time, NDM was the only gene found in all groups (11-28%). NDM metallobeta-lactamase gene was associated with rmpA and iutA genes, giving an example of the connection between virulence and resistance properties. A significant amount of resistant isolates from healthy donors and surface waters indicates the need for additional study of the role of NDM positive isolates in pathogenicity of Klebsiella pneumoniae.

Genomic Characterization of a Proteus sp. Strain of Animal Origin Co-Carrying blaNDM-1 and lnu(G).

The emergence of carbapenem-resistant Proteus represents a serious threat to global public health due to limited antibiotic treatment options. Here, we characterize a Proteus isolate NMG38-2 of swine origin that exhibits extensive drug resistance, including carbapenems. Whole-genome sequencing based on Illumina and MinION platforms showed that NMG38-2 contains 24 acquired antibiotic resistance genes and three plasmids, among which, pNDM_NMG38-2, a pPvSC3-like plasmid, is transferable and co-carries blaNDM-1 and lnu(G). Sequence analysis of pPvSC3-like plasmids showed that they share a conserved backbone but have a diverse accessory module with complex chimera structures bearing abundant resistance genes, which are facilitated by transposons and/or homologous recombination. The acquisition of blaNDM-1 in pNDM_NMG38-2 was due to the ISCR1-mediated integration event. Comprehensive analysis of the lnu(G)-bearing cassettes carried by bacterial plasmids or chromosomes revealed a diversification of its genetic contexts, with Tn6260 and ISPst2 elements being the leading contributors to the dissemination of lnu(G) in Enterococcus and Enterobacteriaceae, respectively. In conclusion, this study provides a better understanding of the genetic features of pPvSC3-like plasmids, which represent a novel plasmid group as a vehicle mediating the dissemination of blaNDM-1 among bacteria species. Moreover, our results highlight the central roles of Tn6260 and ISPst2 in the spread of lnu(G).

A Potential High-Risk Clone of Pseudomonas aeruginosa ST463.

Pseudomonas aeruginosa is one of the most common opportunistic pathogens, which causes severe nosocomial infections because of its well-known multidrug-resistance and hypervirulence. It is critical to curate routinely the epidemic P. aeruginosa clones encountered in the clinic. The aim of the present study was to investigate the connection between virulence factors and antimicrobial resistance profiles in epidemic clones. Herein, we found that ST463 (O4), ST1212 (O11), and ST244 (O5) were prevalent in 30 isolates derived from non-cystic fibrosis patients, based on multilocus sequence type (MLST) and serotype analysis. All isolates were multidrug-resistant (MDR) and each was resistance to at least three classes of antibiotics in antimicrobial susceptibility tests, which was consistent with the presence of the abundant resistance genes, such as blaOXA–50, blaPAO, aph(3′), catB7, fosA, crpP, and blaKPC–2. Notably, all blaKPC–2 genes were located between ISKpn6-like and ISKpn8-like mobile genetic elements. In addition, classical exotoxins encoded by exoU, exoS, and pldA were present in 43.44% (13/40), 83.33% (25/30), and 70% (21/30) of the isolates, respectively. The expression of phz operons encoding the typical toxin, pyocyanin, was observed in 60% of isolates (18/30) and was quantified using triple quadrupole liquid chromatograph mass (LC/MS) assays. Interestingly, compared with other MLST types, all ST463 isolates harbored exoU, exoS and pldA, and produced pyocyanin ranging from 0.2 to 3.2 μg/mL. Finally, we evaluated the potential toxicity of these isolates using hemolysis tests and Galleria mellonella larvae infection models. The results showed that ST463 isolates were more virulent than other isolates. In conclusion, pyocyanin-producing ST463 P. aeruginosa, carrying diverse virulence genes, is a potential high-risk clone.

Discovery and analysis of NBS-LRR gene family in sweet potato genome

<p indent=0mm>The NBS-LRR gene families possess the most abundant resistance genes in plants. Members of the NBS-LRR gene families contain nucleotide-binding site (NBS) and leucine-leucine repeat (LRR) domains. The genome of sweet potato (<italic>Ipomoea batatas</italic>) cultivar has been sequenced but the genes have not been annotated yet. In this study, we predicted the exons of sweet potato genome and obtained the proteins sequences, which were then used to identify and analyze genes of NBS-LRR family. There were 379 genes within NBS-LRR family, amounting to 0.212% of the total genes of sweet potato. The number of the N type, NL type, CNL type, TNL type and PN type was 120, 103, 133, 22, and 1, respectively. All of the chromosomes had NBS-LRR family genes but varied in number and 60.9% of them were clustered. NBS-LRR genes included 15 conservative domains and the genes were conservative within N terminal domain. The results provide references for further studies on the function of NBS-LRR family genes and resistance breeding of sweet potato.

Occurrence and distribution of tetracycline resistance determinants and their pollution profile in the aquaculture environment in Sri Lanka

Tetracyclines (TET) have been extensively used in aquaculture for chemotherapy against various fish diseases. The occurrence of tetracycline contamination levels in the aquaculture farms in Sri Lanka were analyzed using High Performance Liquid Chromatography (HPLC). The presence of antibiotic-resistant bacteria was identified using the 16SrRNA gene sequencing and the corresponding antibiotic resistance genes (ARGs) were screened at each location using PCR method. The concentration of TET in aquaculture farms ranged between 0.001± 0.001 - 0.112± 0.017 ppm whereas Bacillus and Staphylococcus were the most dominant bacterial genera as TET resistant bacteria. Acinetobacter sp., Achromabacter sp., Staphylococcus sp., Micrococcus sp. were also identified as the resistant bacteria. In the present study, tet (M) and tet (A) were abundant resistance genes (11/16 isolates), followed by tet (S) (5/16) and tet (B) (4/16). It was found that tet (M) is one of the most widely distributed tetracycline resistance determinant in the aquatic environment in Sri Lanka.

Prevalence of Antibiotic Resistance Genes and Their Association with Antibiotics in a Wastewater Treatment Plant: Process Distribution and Analysis

Effluents from wastewater treatment plants has been identified as a main point-source of antibiotics and antibiotic resistance genes in natural water environments. In this study, a typical municipal sewage treatment system in south China was taken as the research object to investigate the effects of each treatment unit on eight target antibiotics (sulfamethoxazole, sulfamethazine, tetracycline hydrochloride, oxytetracycline dihydrate, norfloxacin, ofloxacin, clarithromycin, roxithromycin), 17 antibiotic resistance genes (ARGs) and class 1 integron genes in the system using Accelerated Solvent Extraction-Solid phase extraction-Ultra high Performance Liquid Chromatography-Tandem mass spectrometry (ASE-SPE-UPLC-MS/MS) and real-time fluorescent quantitative PCR (qPCR) and the correlation between them. Seven antibiotics (mainly sulfonamides and tetracyclines, 4.19–141.97 ng·L−1) were detected in the influent, while only sulfamethoxazole, sulfamethazine, ofloxacin, and clarithromycin were detected in the effluent (3.11–16.61 ng·L−1). The tetracycline antibiotics in the wastewater treatment plant (WWTP) were transferred to the sludge phase by adsorption, in which tetracycline hydrochloride and oxytetracycline dihydrate were mostly removed in the aerobic and anaerobic stages, while sulfamethoxazole was mainly removed through biological transformation. Sul I was the most abundant resistance gene, but the WWTP had no obvious effect on its removal. Anaerobic treatment was found to play an important role in tetA, tetQ, and tetX removal. Moreover, correlation analysis revealed that the relative abundance of tetX was significantly correlated with clarithromycin (p = 0.039) and ofloxacin (p = 0.028), while that of tetQ was significantly correlated with sulfamethazine (p = 0.007) and sulfamethoxazole (p = 0.001), and that of tetC was significantly correlated with the class 1 integron gene (p = 0.014). Overall, the results presented herein provide a reference for improving the antibiotics and ARGs removal efficiency of WWTPs in south China.

Insights into ecological role of a new deltaproteobacterial order Candidatus Acidulodesulfobacterales by metagenomics and metatranscriptomics

Several abundant but yet uncultivated bacterial groups exist in extreme iron- and sulfur-rich environments, and the physiology, biodiversity, and ecological roles of these bacteria remain a mystery. Here we retrieved four metagenome-assembled genomes (MAGs) from an artificial acid mine drainage (AMD) system, and propose they belong to a new deltaproteobacterial order, Candidatus Acidulodesulfobacterales. The distribution pattern of Ca. Acidulodesulfobacterales in AMDs across Southeast China correlated strongly with ferrous iron. Reconstructed metabolic pathways and gene expression profiles showed that they were likely facultatively anaerobic autotrophs capable of nitrogen fixation. In addition to dissimilatory sulfate reduction, encoded by dsrAB, dsrD, dsrL, and dsrEFH genes, these microorganisms might also oxidize sulfide, depending on oxygen concentration and/or oxidation reduction potential. Several genes with homology to those involved in iron metabolism were also identified, suggesting their potential role in iron cycling. In addition, the expression of abundant resistance genes revealed the mechanisms of adaptation and response to the extreme environmental stresses endured by these organisms in the AMD environment. These findings shed light on the distribution, diversity, and potential ecological role of the new order Ca. Acidulodesulfobacterales in nature.

Metagenomic Insights Into the Contribution of Phages to Antibiotic Resistance in Water Samples Related to Swine Feedlot Wastewater Treatment.

In this study, we examined the types of antibiotic resistance genes (ARGs) possessed by bacteria and bacteriophages in swine feedlot wastewater before and after treatment using a metagenomics approach. We found that the relative abundance of ARGs in bacterial DNA in all water samples was significantly higher than that in phages DNA (>10.6-fold), and wastewater treatment did not significantly change the relative abundance of bacterial- or phage-associated ARGs. We further detected the distribution and diversity of the different types of ARGs according to the class of antibiotics to which they confer resistance, the tetracycline resistance genes were the most abundant resistance genes and phages were more likely to harbor ATP-binding cassette transporter family and ribosomal protection genes. Moreover, the colistin resistance gene mcr-1 was also detected in the phage population. When assessing the contribution of phages in spreading different groups of ARGs, β-lactamase resistance genes had a relatively high spreading ability even though the abundance was low. These findings possibly indicated that phages not only could serve as important reservoir of ARG but also carry particular ARGs in swine feedlot wastewater, and this phenomenon is independent of the environment.

Impact of Wastewater Treatment on the Prevalence of Integrons and the Genetic Diversity of Integron Gene Cassettes.

The integron platform allows the acquisition, expression, and dissemination of antibiotic resistance genes within gene cassettes. Wastewater treatment plants (WWTPs) contain abundant resistance genes; however, knowledge about the impacts of wastewater treatment on integrons and their gene cassettes is limited. In this study, by using clone library analysis and high-throughput sequencing, we investigated the abundance of class 1, 2, and 3 integrons and their corresponding gene cassettes in three urban WWTPs. Our results showed that class 1 integrons were most abundant in WWTPs and that wastewater treatment significantly reduced the abundance of all integrons. The WWTP influents harbored the highest diversity of class 1 integron gene cassettes, whereas class 3 integron gene cassettes exhibited highest diversity in activated sludge. Most of the gene cassette arrays detected in class 1 integrons were novel. Aminoglycoside, beta-lactam, and trimethoprim resistance genes were highly prevalent in class 1 integron gene cassettes, while class 3 integrons mainly carried beta-lactam resistance gene cassettes. A core class 1 integron resistance gene cassette pool persisted during wastewater treatment, implying that these resistance genes could have high potential to spread into environments through WWTPs. These data provide new insights into the impact of wastewater treatment on integron pools and highlight the need for surveillance of resistance genes within both class 1 and 3 integrons.IMPORTANCE Wastewater treatment plants represent a significant sink and transport medium for antibiotic resistance bacteria and genes spreading into environments. Integrons are important genetic elements involved in the evolution of antibiotic resistance. To better understand the impact of wastewater treatment on integrons and their gene cassette contexts, we conducted clone library construction and high-throughput sequencing to analyze gene cassette contexts for class 1 and class 3 integrons during the wastewater treatment process. This study comprehensively profiled the distribution of integrons and their gene cassettes (especially class 3 integrons) in influents, activated sludge, and effluents of conventional municipal wastewater treatment plants. We further demonstrated that while wastewater treatment significantly reduced the abundance of integrons and the diversity of associated gene cassettes, a large fraction of integrons persisted in wastewater effluents and were consequentially discharged into downstream natural environments.

The antibiotic resistance of heterotrophic bacteria in tap waters in London

Abstract This study assessed the occurrence and prevalence of antibiotic-resistant bacteria (ARBs) and antibiotic resistance genes (ARGs) in tap water sampled across London, United Kingdom. Sampling was conducted seasonally from nine locations spread geographically across the city. ARBs and ARGs (tet(A), dfrA7, and sul1) were detected in all sampling locations in all sampling rounds. Resistance to trimethoprim was the highest among the tested antibiotics and the sul1 gene was the most abundant resistance gene detected. Several opportunistic pathogens were identified amongst the ARBs in the water samples, including Pseudomonas aeruginosa and Stenotrophomonas maltophilia.

Contamination profiles of antibiotic resistance genes in the sediments at a catchment scale

The aim of this study was to investigate the contamination profiles of tetracycline, sulfonamide, and macrolide resistance genes, as well as integrons in sediments of Dongjiang River basin of South China by real time quantitative polymerase chain reaction. sul2 was the most abundant resistance gene, with the average concentration of 6.97×108 copies/g and 1.00×108 copies/g in the dry and wet seasons, respectively, followed by ermF, sul3, sul1, intI1, tetA, ermB, tetX, tetM, tetQ, tetO, tetW, tetS, ermC, and tetB. The abundance of intI2 gene was the lowest in the sediment samples. Significant correlations existed between the ARGs and sediment properties as well as metals (Cu and Zn) and corresponding antibiotic classes, suggesting that the contamination of ARGs is related to chemical pollution of the sediments in the river basin. Principal component analysis showed distinct groupings of the sampling sites, reflecting that human activities are the key player in the dissemination of ARGs in the catchment environment.