Occurrence Of Antibiotic Resistance Genes Research Articles

Prediction of antibiotic-resistance genes occurrence at a recreational beach with deep learning models

Antibiotic resistance genes (ARGs) have been reported to threaten the public health of beachgoers worldwide. Although ARG monitoring and beach guidelines are necessary, substantial efforts are required for ARG sampling and analysis. Accordingly, in this study, we predicted ARGs occurrence that are primarily found on the coast after rainfall using a conventional long short-term memory (LSTM), LSTM-convolutional neural network (CNN) hybrid model, and input attention (IA)-LSTM. To develop the models, 10 categories of environmental data collected at 30-min intervals and concentration data of 4 types of major ARGs (i.e., aac(6′-Ib-cr), blaTEM, sul1, and tetX) obtained at the Gwangalli Beach in South Korea, between 2018 and 2019 were used. When individually predicting ARGs occurrence, the conventional LSTM and IA-LSTM exhibited poor R2 values during training and testing. In contrast, the LSTM-CNN exhibited a 2–6-times improvement in accuracy over those of the conventional LSTM and IA-LSTM. However, when predicting all ARGs occurrence simultaneously, the IA-LSTM model exhibited a superior performance overall compared to that of LSTM-CNN. Additionally, the influence of environmental variables on prediction was investigated using the IA-LSTM model, and the ranges of input variables that affect each ARG were identified. Consequently, this study demonstrated the possibility of predicting the occurrence and distribution of major ARGs at the beach based on various environmental variables, and the results are expected to contribute to management of ARG occurrence at a recreational beach.

Upraising a silent pollution: Antibiotic resistance at coastal environments and transference to long-distance migratory shorebirds.

Large amounts of antibiotics from different sources have been released into coastal environments, especially in high human-populated areas, but comprehensive studies of antibiotic footprint in wildlife are scarce. Here we assess occurrence of antibiotic resistant bacteria (ARB) and antibiotic resistance gene (ARG) both in sediments and gut microbiota of a long-distance migratory shorebird species in two coastal wetlands at a sparsely-populated area in Pacific Patagonian coasts with contrasting potential antibiotic sources, especially from aquaculture. We found 62% of sediment samples showing ARB, and ARGs similarly occurring in sediments at both bays. However multi-resistant ARB were found only at sediments in the bay surrounding aquaculture operations. An 87% of cloacal bird samples showed at least one ARB, with 63% being multi-resistant and some of them with a high potential pathogenicity. ARGs were present in 46% of the samples from birds, with similar multi-resistant frequencies among bays. Besides specific differences mainly associated to antibiotics used in salmon aquaculture that boosted ARB in sediments, ARB and ARGs occurrence was overall similar at two bays with contrasting main human activities, in spite of being a comparatively low human-populated area. Therefore, our results reinforce the idea that the antibiotic footprint may be widespread at a global scale and can extend beyond the geographical influence of antibiotic sources, especially at coastal environments where migratory shorebirds act both as reservoirs and potential spreaders of antibiotic resistance.

Response of antibiotic and heavy metal resistance genes to tetracyclines and copper in substrate-free hydroponic microcosms with Myriophyllum aquaticum

Constructed wetlands for antibiotics and heavy metals removal have become important reservoirs of antibiotic resistance genes (ARGs) and heavy metal resistance genes (MRGs), especially in the substrates. Here, substrate-free hydroponic microcosms of Myriophyllum aquaticum were established; tetracyclines (TCs) and Cu(II) were added to evaluate the behaviours of ARGs and MRGs in the microcosms. Several ARGs, MRGs, and mobile genetic elements (MGE) were detected in the biofilms attached to the plants, ranging from 0.5 to 2.3 × 108 copies/g dry weight. ARGs and MRGs exhibited higher relative abundances in the effluent suspended solids (SS); however, their absolute amounts were much lower than those in conventionally constructed wetlands. Microcosms with TCs and Cu(II) exhibited a higher level of resistant genes than those with compound added singularly owing to co-selection pressure. The existence of TCs and copper significantly changed the microbial communities in the microcosms. The exogenous input of TC/Cu(II) and microbial community structure were the factors driving the occurrence of ARGs, whereas MRGs were more correlated with the copper addition. Thus, reducing the exogenous inputs of antibiotics /heavy metals and SS of the effluent is suggested for the mitigation of resistant genes in phytoremediation technologies working in the absence of conventional substrates.

Bacterial communities, potential pathogens and antibiotic resistance genes of silver-loaded stainless steel pipe wall biofilm in domestic hot water system

Silver-loaded stainless steel (Ag-SS) was applied in domestic hot water system (DHWS) to control the biofilm contamination. The antibacterial properties of Ag-SS, as well as the bacterial communities, potential pathogens and antibiotic resistance genes (ARGs) of pipe wall biofilm were investigated, with SS as control. Ag-SS maintained a high antibacterial efficiency on heterotrophic bacteria (70.60 ± 13.22 %) and Escherichia coli (73.56 ± 10.62 %) by contact bacteriostasis. Dominant phyla exhibited a lower relative abundance in Ag-SS than SS. The bacteria were further classified into abundant (≥ 1 %), moderately abundant (0.1∼1 %) and rare (≤ 0.1 %) taxa to investigate their responses to silver. There were 15.33 % of shared operational taxonomic units (OTUs) belonged to abundant taxa, while most unique OTUs (> 80 %) of each sample were identified as rare taxa. As the primary abundant genera, Mesorhizobium and Bradyrhizobium exhibited resistant and sensitive to silver, respectively. Most dominant moderately abundant and rare genera were resistant to silver. Moreover, among 19 detected potential pathogens, Legionella, Parachlamydia and Aeromonas were sensitive to silver, while Pseudomonas and Acinetobacter were resistant to silver. Furthermore, a total of 48 ARGs were detected and they exhibited similar distributions in both samples. The total abundance of ARGs in Ag-SS increased by 0.08 fold compared with that in SS, with efflux pumps as the main resistance mechanisms. This study highlights the different responses of abundant, moderately abundant and rare taxa to silver, and the occurrences of ARGs, confirming that Ag-SS was an effective disinfection strategy for biosafety protection of DHWS.

Shifts in bacterial communities and antibiotic resistance genes in surface water and gut microbiota of guppies (Poecilia reticulata) in the upper Rio Uberabinha, Brazil

Anthropogenic activities especially water pollution can affect the diversity and composition of microbial communities and promote the spread of antibiotic resistance genes (ARGs). In this study, water samples and guppies (Poecilia reticulata) were sampled from six sampling sites along the Uberabinha River in southeastern Brazil, both microbial communities and ARGs of surface waters and intestinal microbiota of guppies (Poecilia reticulata) were detected. According to the results of 16S rRNA amplicon sequencing, Proteobacteria, Bacteroidetes, Firmicutes and Actinobacteria were dominant phyla in both water and intestinal microbiota, but the abundance of putative pathogens was higher at heavily polluted sites. Up to 83% of bacteria in intestinal microbiota originated from water microbiota; this proportion was relatively higher in less polluted compared to polluted environments. ARGs providing resistance of tetracyclines and quinolones were dominant in both water and gut microbiota. The relative abundances of class I integrons and ARGs were as high as 1.74 × 10−1/16S rRNA copies and 3.61 × 10−1/16S rRNA copies, respectively, at heavily polluted sites. Correlation analysis suggests that integrons and bacteria play key roles in explaining the widespread occurrence of ARGs in the surface, but not in intestinal microbiota. We could rule out the class I integrons a potential intermediary bridge for ARGs between both types of microbiomes. Our results highlight the tight link in microbial communities and ARGs between ambient microbiota of stream ecosystems and intestinal microbiota of fish. Our study could have far-reaching consequences for fisheries and consumer safety and calls for investigations of gut microbiota of target species of both commercial fisheries and recreational (hobby) angling.

Effects of polyethylene microplastics on the fate of antibiotic resistance genes and microbial communities in anaerobic digestion of dairy wastes

Microplastics (MPs) have been confirmed to affect the digestion performance of the anaerobic digestion (AD) and to influence the occurrence of antibiotic resistance genes (ARGs) in the environment. In this study, the effects of polyethylene (PE) MPs on digestion performance and the fate of ARGs and microbial communities in thermophilic AD (55 °C) and hyperthermophilic AD (65 °C) of dairy wastes were investigated. The results showed that the effects of PE MPs were similar at different temperatures. At 55 °C and 65 °C, the presence of PE MPs improved methane production by 8.4% and 41.2% and enhanced COD removal rates to 52.8% and 52.4%, respectively, mainly by increasing the abundances of microorganisms involved in hydrolysis, acidogenesis and acid utilization (e.g. Clostridium , Dechloromonas and Caldicoprobacter ). PE MPs significantly increased the fold changes in the abundances of tet C, tet G and tet W from 0.71 to 12.63 to 1.15–28.62. According to redundancy analysis and correlation analysis, free ammonia contributed the most (51% of the total variation) to the changes in host bacteria, which probably changed the fate of ARGs in different AD system. This study indicated the discharge of PE MPs presents a novel challenge in reducing the risks of ARG transmission. • The effects of polyethylene microplastics on anaerobic digestion were investigated. • Polyethylene efficiently increased the methane production and COD removal rate. • Polyethylene significantly increased the abundances of tet C, tet G and tet W. • Polyethylene shifted the microbial communities involving gas production.

Occurrence of Antibiotic Resistance Genes and Bacterial Community Structure of Different Sludge Samples During Microwave Pretreatment-Anaerobic Digestion

The waste sludge of municipal wastewater treatment plants is an important reservoir for antibiotic resistance genes (ARG). It is necessary to explore the fate of ARG, microbial community succession, and the correlations between them. Therefore, the distribution of ARG and the microbial community structure of waste sludge from wastewater treatment plants with A2O and A2O-MBR processes during microwave pretreatment and anaerobic digestion were studied in this research. The results showed that the occurrence of ARG and the microbial community structure were quite different in the waste sludge of A2O and A2O-MBR processes. The microwave pretreatment did not change the microbial community much, whereas the community structure of the digested sludge with pretreatment showed significant differences. Anaerobic digestion had a conformity effect on the distribution of ARG and MGE in the digested sludge with or without pretreatment. Among genes, ermF, qnrS, and blaNDM-1 were the most difficult to be reduced ARG and were prone to propagation during anaerobic digestion. The influence of biomass, ammonia nitrogen, and phosphorus on the distribution of ARG and MGE was higher than that of other environmental factors. The sludge characteristics also showed important impacts on the microbial community, especially on some genera with specific functions. These results could help people to better understand the spread and control of ARG during sludge anaerobic digestion.

Determination of antibiotic resistance genes in a WWTP-impacted river in surface water, sediment, and biofilm: Influence of seasonality and water quality

Many pathogenic bacteria are adapted to live in aquatic habitats, which makes rivers possible sources and spread pathways of antibiotic resistance, since they usually receive effluents from wastewater treatment plants (WWTP), possibly containing antibiotic residues and also antibiotic-resistant bacteria. This study investigates different monitoring strategies to identify the occurrence of antibiotic-resistant bacteria in rivers. We analyzed the presence of 13 antibiotic resistance genes (ARGs) and seven gene markers for facultative pathogenic bacteria (FPB) with qPCR in sampling sites upstream and downstream of a small WWTP in Southern Germany. Five sampling campaigns were conducted from February to June 2019. Surface water, sediment, and biofilm samples were analyzed. The biofilm was collected from an artificial sampler placed in the river. blaTEM, ermB, tetM, and sul1 genes were detected in all samples analyzed. The results showed there was a previous background in the river, but the WWTP and the water quality of the river influenced the concentration and occurrence of ARGs and FPB. Genes representing resistance against strong or last-resort antibiotics, such as mecA, blaCMY-2, blaKPC-3, and mcr-1, and multidrug resistance were also detected, mainly in samples collected downstream of the WWTP. Downstream of the WWTP, the occurrence of ARG and FPB correlated with ammoniacal nitrogen, while upstream of the WWTP correlated with turbidity, suspended solids, and seasonal factors such as UVA radiation and the presence of macrophytes. Biofilm samples presented higher abundances of ARGs and FPB. The biofilm sampler was efficient and allowed to collect biofilms from specific periods, which helped to identify seasonal patterns.

Selection of antibiotic resistance genes on biodegradable and non-biodegradable microplastics

Growing evidence have demonstrated that microplastics in the marine ecosystem can provide novel substrates for biofilm formation, potentially facilitating the spread of antibiotic resistance. However, the occurrence of antibiotic resistance genes (ARGs) in the biofilm on microplastics has not been fully explored. This study used the metagenomic data of biodegradable and non-biodegradable microplastics staged at a coastal lagoon in the northern Gulf of Mexico to profile the ARGs and their bacterial hosts. The abundance and Shannon diversity of ARGs on biodegradable poly hydroxy alkanoate (PHA) and non-biodegradable polyethylene terephthalate (PET) have no significant differences. Nevertheless, the abundance of multidrug resistance genes on PET (3.05 copies per 16S rRNA) was statistically higher than that on PHA (2.05). Beta diversity showed that the overall pattern of resistome on PHA was significantly distinct with that on PET. Procrustes analysis suggested a good-fit correlation between ARG profiles and bacterial community composition. The host-tracking analysis identified that Pseudomonas was always the major host for glycopeptide and multidrug resistance genes in PET and PHA biofilms, whereas the primary host for macrolide-lincosamide-streptogramin (MLS) changed to Desulfovibrio on PET. This study provided the first metagenomic insights into the ARGs and their hosts on biodegradable and non-biodegradable microplastics, suggesting that both two types of plastics harbor ARGs with preferences.

Accumulation of antibiotic resistance genes in full-scale drinking water biological activated carbon (BAC) filters during backwash cycles

Biological activated carbon (BAC) filtration, a process widely used in drinking water treatment, was recently reported to harbor antibiotic resistance genes (ARGs). This emerging contamination is poorly understood. This study was conducted to investigate the occurrence of ARGs and bacterial community in full-scale BAC filters during the backwash cycle using high-throughput qPCR and high-throughput sequencing. A total of 178 ARGs were detected in all biofilm samples, with relative abundance ranging from 0.1 to 1.37 copies per 16S rRNA and absolute abundance ranging from 4.48 × 107 to 3.09 × 109 copies/g carbon. Biofilms sampled from different filters shared most detected ARGs and dominant genera including Bryobacter, Pedomicrobium, Reyranella, and Terrimonas, though their bacterial community structure differed significantly. After backwashing, the relative ARGs abundance increased by 1.5- to 3.8-folds and the absolute ARGs abundance increased by 0.90- to 1.12-logs in all biofilm samples during filter ripening, indicating that ARGs accumulated in filters during this period. Redundancy analysis suggested that such ARGs accumulation was mainly driven by horizontal gene transfer in winter, but highly correlated with the increasing relative abundance of genera Bryobacter and Acidibacter in summer. It was observed that 80.6 %-89.3% of the detected ARGs persisted in the filters despite of the backwashing. Given the high richness and relative abundance of ARGs in BAC filter and the ineffectiveness of backwashing in ARG removal, more stringent downstream disinfection strategies are deserved and more research is necessary to assess potential human health risks due to the persistence of ARGs in drinking water.

Microplastics act as vectors for antibiotic resistance genes in landfill leachate: The enhanced roles of the long-term aging process.

Microplastics (MPs) are found to be ubiquitous and serve as vectors for other contaminants, and the inevitable aging process changes MP properties and fates. However, whether the MPs in aging process affects the fates of antibiotic resistance gene (ARGs) in aquatic environments is poorly understood. Herein, the physicochemical property alteration of MPs being aged in landfill leachate, an important reservoir of MPs and ARGs, was investigated, and microbial community evolution and ARGs occurrence of MP surface during the aging process were analyzed. Aging process remarkably altered surface properties, including increasing specific surface areas, causing the formation of oxygen-containing groups, and changing surface morphology, which further increased the probability of microbial colonization. The bacterial assemblage on MPs showed higher biofilm-forming and pathogenic potential compared to leachate. ARGs quantification results suggested that MPs exhibited selective enrichment for ARGs in a ratio of 5.7-103 folds, and the aging process enhanced the enrichment potential. Further co-occurrence networks suggested that the existence of non-random, closer and more stable ARGs-bacterial taxa relations on MP surface affected the ARG transmission. The study of ARG partitioning on MPs indicated that extracellular DNA was a nonnegligible reservoir of ARGs attached on MP surface, and that biofilm bacterial community influenced ARGs partitioning pattern during the aging process. This study confirmed that the aging process could enhance the potential of MPs as vectors for ARGs, which would promote the holistic understanding of MP behavior and risk in natural environments.

Reponses of microbial community and antibiotic resistance genes to the selection pressures of ampicillin, cephalexin and chloramphenicol in activated sludge reactors

High concentrations of antibiotics can exert strong selection pressures on the microbial community and promote the emergence and dissemination of antibiotic resistance genes (ARGs). The activated sludge reactors treating ampicillin, cephalexin and chloramphenicol production wastewater were established to investigate the responses of microbial community, ARGs and mobile genetic elements (MGEs) to antibiotics. Antibiotic selection pressures significantly declined the microbial diversity and changed microbial community structures. Based on metagenomic analysis, a total of 500 ARG subtypes affiliated with 18 ARG types were identified and 63 ARGs were shared by all samples. The substantial increase of ARG abundance and the shifts of ARG profiles were significantly correlated with antibiotic types and concentrations. The evident enrichment of non-corresponding ARG types suggested the strong co-selection effects of the target antibiotics. Additionally, metagenomic analysis revealed the occurrence of 104 MGEs belonging to various types and the five dominant MGEs were tnpA, intI1, tniA, tniB and IS91. The ARG–MGE co-occurrence associations implied the potential mobility of ARGs. Network analysis also demonstrated that five ARG types (aminoglycoside, beta-lactam, chloramphenicol, multidrug and tetracycline resistance genes) tended to co-occur internally and the obvious co-occurrence patterns among different ARG types indicated the potential for resistance co-selection. Moreover, 15 bacterial genera were speculated as the hosts of diverse ARGs. This study provides a comprehensive overview of the occurrence of ARGs and MGEs and is valuable for the risk assessment and management of antibiotic resistance.

Horizontal and vertical gene transfer drive sediment antibiotic resistome in an urban lagoon system

Rapid urbanization has resulted in pervasive occurrence of antibiotic resistance genes (ARGs) in urban aquatic ecosystems. However, limited information is available concerning the ARG profiles and the forces responsible for their assembly in urban landscape lagoon systems. Here, we employed high-throughput quantitative PCR (HT-qPCR) to characterize the spatial variations of ARGs in surface and core sediments of Yundang Lagoon, China. The results indicated that the average richness and absolute abundance of ARGs were 11 and 53 times higher in the lagoon sediments as compared to pristine reference Tibetan lake sediments, highlighting the role of anthropogenic activities in ARG pollution. Co-occurrence network analysis indicated that various anaerobic prokaryotic genera belonging to Alpha-, Deltaproteobacteria, Bacteroidetes, Euryarchaeota, Firmicutes and Synergistetes were the potential hosts of ARGs. The partial least squares-path modeling (PLS-PM) analysis revealed positive and negative indirect effects of physicochemical factors and heavy metals on the lagoon ARG profiles, via biotic factors, respectively. The horizontal (mediated by mobile genetic elements) and vertical (mediated by prokaryotic communities) gene transfer may directly contribute the most to drive the abundance and composition of ARGs, respectively. Furthermore, the neutral community model demonstrated that the assembly of sediment ARG communities was jointly governed by deterministic and stochastic processes. Overall, this study provides novel insights into the diversity and distribution of ARGs in the benthic habitat of urban lagoon systems and underlying mechanisms for the spread and proliferation of ARGs.

Response of Cyperus involucratus to sulfamethoxazole and ofloxacin-contaminated environments: Growth physiology, transportation, and microbial community

Plant-microbe is a complementary coupling system for antibiotics removing in constructed wetlands (CWs), but how plant and rhizosphere microbiomes respond to antibiotics exposure and the occurrence of ARGs in this microenvironment have seldom been researched. Thus, the response of the plant-microbe coupling system to different levels of antibiotics (sulfamethoxazole (SMZ) and ofloxacin (OFL)) was investigated. The results showed that two antibiotic stressors have hormetic effects on plant growth, physiology, and microbial community evolution, and the antibiotic toxic effects presented as SMZ + OFL > SMZ > OFL. Antibiotic accumulation in the plants was in the order of roots > stems > leaves. Notably, the root attachments affected antibiotic transportation. The accumulation of antibiotics in the under-ground parts affected the rhizosphere microbial community structure, and the microorganisms were more sensitive to SMZ + OFL than the plants, with inflection points of 0.5 mg L−1 and 1 mg L−1, respectively. Pseudomonas was highly resistant to antibiotics, while Acidovorax and Devosia may play a role in antibiotic degradation. Correlation analysis and network analysis showed that antibiotic enrichment and the bacterial community contributed significantly to the abundance of antibiotic-resistant genes (ARGs), further revealing the co-occurrence of int1, ARGs, and the potential bacterial hosts.

The Toothbrush Microbiome: Impact of User Age, Period of Use and Bristle Material on the Microbial Communities of Toothbrushes.

Toothbrushes play a central role in oral hygiene and must be considered one of the most common articles of daily use. We analysed the bacterial colonization of used toothbrushes by next generation sequencing (NGS) and by cultivation on different media. Furthermore, we determined the occurrence of antibiotic resistance genes (ARGs) and the impact of different bristle materials on microbial growth and survival. NGS data revealed that Enterobacteriaceae, Micrococcaceae, Actinomycetaceae, and Streptococcaceae comprise major parts of the toothbrush microbiome. The composition of the microbiome differed depending on the period of use or user age. While higher fractions of Actinomycetales, Lactobacillales, and Enterobacterales were found after shorter periods, Micrococcales dominated on both toothbrushes used for more than four weeks and on toothbrushes of older users, while in-vitro tests revealed increasing counts of Micrococcus on all bristle materials as well. Compared to other environments, we found a rather low frequency of ARGs. We determined bacterial counts between 1.42 × 106 and 1.19 × 107 cfu/toothbrush on used toothbrushes and no significant effect of different bristles materials on bacterial survival or growth. Our study illustrates that toothbrushes harbor various microorganisms and that both period of use and user age might affect the microbial composition.

Spatial distribution of antibiotic and heavy metal resistance genes in the Black Sea

Antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB) are worldwide considered as emerging contaminants of large interest, and a primary threat to human health. It is becoming clear that the environment plays a central role in the transmission, spread, and evolution of antibiotic resistance. Although marine systems have been largely investigated, only a few studies have considered the presence of ARGs in meso- and bathypelagic waters. To date, no molecular based studies have yet been made to investigate the occurrence of ARGs in the Black Sea, the largest meromictic basin in the world, receiving water from a number of important European rivers and their residues of anthropogenic activities in permanently stratified mesopelagic water masses. In this study, we determined the presence and the abundance of five ARGs (blaCTXM, ermB, qnrS, sul2, tetA) and of the heavy metal resistance gene (HMRG) czcA, in different sampling sites in the eastern and western Black Sea, at several depths (up to 1000 m) and various distances from the shoreline. Three ARGs (blaCTXM, sul2, and tetA) and czcA were present in at least 43% of the analysed samples, whereas ermB and qnrS were never detected. In particular, sul2 abundances increased significantly in coastal location, whereas tetA increased with sampling depth. These findings point out the Black Sea as a source of ARGs and HMRGs distributed along the whole water column.

Abundance, diversity and mobility potential of antibiotic resistance genes in pristine Tibetan Plateau soil as revealed by soil metagenomics.

Widespread occurrence of antibiotic resistance genes (ARGs) has become an important clinical issue. Studying ARGs in pristine soil environments can help to better understand the intrinsic soil resistome. In this study, 10 soil samples were collected from a high elevation and relatively pristine Tibetan area, and metagenomic sequencing and bioinformatic analyses were conducted to investigate the microbial diversity, the abundance and diversity of ARGs and the mobility potential of ARGs as indicated by different mobile genetic elements (MGEs). A total of 48 ARG types with a relative abundance of 0.05-0.28 copies of ARG/copy of 16S rRNA genes were detected in Tibetan soil samples. The observed ARGs were mainly associated with antibiotics that included glycopeptide and rifamycin; the most abundant ARGs were vanRO and vanSO. Low abundance of MGEs and potentially plasmid-related ARGs indicated a low horizontal gene transfer risk of ARGs in the pristine soil. Pearson correlation and redundancy analyses showed that temperature and total organic carbon were the major environmental factors controlling both microbial diversity and ARG abundance and diversity.

Occurrence and distribution of antibiotic resistance genes in lakes and reservoirs from water-receiving area of Eastern Route of the South-to-North Water Diversion Project, Northern China

Abstract To characterize antibiotic resistance genes (ARGs) in surface waterbodies in Shandong Province, and to understand the occurrence, diversity and resistance mechanism of ARGs, nine aquatic ecosystems were selected and evaluated regarding occurrence of ARGs in the period from 2018 to 2019. Thirty-three types and 242 subtypes of ARGs were detected in the nine waterbodies with the most subtypes in Wohushan (242) and Jihongtan (235), and the fewest ARGs in Datun (213) and Mishan (221). Among the top nine ARG types, MacB, BcrA, CarA and TetW were the dominant ARGs in these samples. Four groups of the study sites were distinguished by cluster analysis, indicating different distribution characteristics of ARGs in these waterbodies, and regional differences in ARG contamination. Efflux pumps were the predominant resistance mechanism in all the waterbodies in terms of both subtype number (49.2%) and abundance (70.4%), followed by operon, antibiotic inactivation and others. Our findings provide comprehensive information to better understand the contamination level of ARGs on a local level as well as for global considerations, and may be practically helpful for the prevention and control of ARG pollution.

Critical roles of cyanobacteria as reservoir and source for antibiotic resistance genes

The widespread occurrence of antibiotic resistance genes (ARGs) throughout aquatic environments has raised global concerns for public health, but understanding of the emergence and propagation of ARGs in diverse environmental media remains limited. This study investigated the occurrence and spatio-temporal patterns of six classes of ARGs in cyanobacteria isolated from Taihu Lake. Tetracycline and sulfonamide resistance genes were identified as dominant ARGs. The abundance of ARGs in cyanobacteria was significantly higher in the bloom period than in the non-bloom period. The contribution and persistence of ARGs were higher in extracellular DNA (eDNA) than in intracellular DNA (iDNA) from cyanobacteria. Cyanobacteria-associated eDNA carrying ARGs was more stable at lower temperature. The relative abundances of ARGs in Microcystis and Synechococcus, the dominant genera of cyanobacterial blooms in Taihu Lake, were significantly higher than those in other cyanobacterial strains. The conjugative transfer efficiency for bacterial assimilation of ARGs in cyanobacteria was facilitated by increasing temperature and cyanobacterial cell concentration. Our results demonstrated that cyanobacteria could act as a significant reservoir and source for the acquisition and dissemination of ARGs in aquatic environments, hence the definition of negative ecological effects of cyanobacterial blooms was expanded.

Antibiotic Resistance Genes Occurrence in Wastewaters from Selected Pharmaceutical Facilities in Nigeria

The proliferation of antibiotic-resistant bacteria (ARB) and the prevalence of antibiotic resistance genes (ARGs) in wastewaters are well-established factors that contribute to the reduced potency of antibiotics used in healthcare worldwide. The human health risk associated with the proliferation of ARB and ARGs need to be understood in order to design mitigation measures to combat their dissemination. Using the PCR analysis of genomic DNA, the prevalence of 41 ARGs active against the commonly used six classes of antibiotics was evaluated in 60 bacterial isolates obtained from pharmaceutical wastewaters in Nigeria. The ARGs most frequently detected from the bacterial isolates in each of the antibiotic classes under study include catA1 (58.3%); sulI (31.7%); tet(E) (30%); aac(3)-IV (28.3%); ermC (20%); blaTEM, blaCTX-M, blaNDM-1 at 18.3% each; which encode for resistance to chloramphenicol, sulfonamides, tetracycline, aminoglycoside, macrolide-lincosamide-streptogramin and β-lactams and penicillins, respectively. Acinetobacter spp., accession number MH396735 expressed the highest number of ARGs of all the bacterial isolates, having at least one gene that encodes for resistance to all the classes of antibiotics in the study. This study highlights wide distribution of ARB and ARGs to the antibiotics tested in the wastewater, making pharmaceutical wastewater reservoirs of ARGs which could potentially be transferred from commensal microorganisms to human pathogens.