Mobile genetic elements (MGEs) such as plasmids or integrative conjugative elements (ICEs) are widely involved in the horizontal transfer of antibiotic resistant genes (ARGs), but their environmental host-range and reservoirs remain poorly known, as mainly assessed through the analysis of culturable and clinical bacterial isolates. In this study, we used a gradual approach for determining the environmental abundance and host-range of ICEs belonging to the SXT/R391 family, otherwise well known to bring ARGs in Vibrio spp. epidemic clones and other pathogens. First, by screening a set of aquatic bacteria libraries covering 1794 strains, we found that almost 1% of the isolates hosted an SXT/R391 element, all belonging to a narrow group of non-O1/non-O139 Vibrio cholerae. However, when SXT/R391 ICEs were then quantified in various aquatic communities, they appeared to be ubiquitous and relatively abundant, from 10⁻⁶ to 10⁻³ ICE copies per 16 S rDNA. Finally, the molecular exploration of the SXT/R391 host-range in two river ecosystems impacted by anthropogenic activities, using the single-cell genomic approach epicPCR, revealed several new SXT/R391 hosts mostly in the Proteobacteria phylum. Some, such as the pathogen Arcobacter cryaerophilus (Campylobacteraceae), have only been encountered in discharged treated wastewaters and downstream river waters, thus revealing a likely anthropogenic origin. Others, such as the non-pathogenic bacterium Neptunomonas acidivorans (Oceanospirillaceae), were solely identified in rivers waters upstream and downstream the treated wastewaters discharge points and may intrinsically belong to the SXT/R391 environmental reservoir. This work points out that not only the ICEs of the SXT/R391 family are more abundant in the environment than anticipated, but also that a variety of unsuspected hosts may well represent a missing link in the environmental dissemination of MGEs from and to bacteria of anthropogenic origin.

Current understanding of biochar's effect on antibiotic resistance genes (ARGs) in soil is limited, and whether the effect could change after long-term field aging remains largely unknown. In this study, we employed high-throughput quantitative PCR to assess the effect of biochar amendment on soil resistome as affected by three years of field aging. Application of fresh biochar significantly elevated the number and abundance of ARGs in the manured soil, but did not show such effect under pakchoi cultivation. The presence of aged biochar caused a marked reduction of ARGs only in the planted manured soil. Results of principal coordinate analysis and structural equation modeling indicate that biochar's effect on soil ARG profile was changed by field aging through altering soil microbial composition. These results highlight the necessity of considering aging effect of biochar during its on-farm application to mitigate soil antibiotic resistance.

Infection caused by pathogenic bacteria carrying antibiotic resistance genes (ARGs) is a serious challenge to human health. Water environment, including water and surface sediments, is an important repository of ARGs, and the activity of aquatic animal can affect the development of ARG pollution in the water environment. Macrobenthic invertebrates are an important component of aquatic ecosystems, and their effects on ARG development in aquatic environments remain unreported. The distribution of ARGs, including tetA gene, sul2 gene, and kan gene, in Chironomidae larvae is demonstrated in this study for the first time. The ARG distribution was related to sampling points, metal elements, and seasons. Animal models demonstrated that Chironomidae larvae enriched ARGs from water and passed them on to downstream predators in the food chain. Conjugative transfer mediated by resistant plasmids was crucial in the spread of ARG in Chironomidae larvae, and upregulated expression of trfAp gene and trbBp gene was the molecular mechanism. Escherichia in Proteobacteria and Flavobacterium in Bacteroidetes, which are gram-negative bacteria in Chironomidae larvae, are the primary host bacteria of ARGs confirmed via resistance screening and DNA sequencing of V4 region of 16S rRNA gene. Feeding experiments further confirmed that ARGs from Chironomidae larvae can be enriched in the fish gut. Research gaps in food chain between sediments and fish are addressed in this study, and Chironomidae larvae is an important enricher of ARGs in the freshwater environment.

Antibiotics in the environment usually co-exist with their transformation products with retained toxicity, raising concerns about environmental risks of their combined exposure. Herein, we reported a novel predictive approach for evaluating the individual and combined toxicity for photodegradation products of fluoroquinolone antibiotics (FQs). Quantitative structure-activity relationship (QSAR) models with promising predictive performance were constructed and validated using experimental data obtained with 13 FQs and 78 mixtures towards E. coli. A structural descriptor reflecting the interaction among FQ molecules and the target protein was employed in the QSAR models, which was obtained through molecular docking and thus provided a rational mechanistic explanation for these models. The predicted results indicated that the degradation products displayed varying degrees of changes compared to the parent FQs, while the combined toxicity of FQs and their degradation products was mostly additive. Furthermore, following UV irradiation the degradation products displayed elevated capacity of inducing resistance mutations in E. coli, though their overall toxicity was reduced. This result highlights the implications of antibiotic degradation products on resistance development in bacteria and stresses the importance of considering such impacts during environmental risk assessments of antibiotics.

Antibiotic resistance is a major public health concern. Triclosan is an antimicrobial compound with direct links to antibiotic resistance that was widely used in soaps in the U.S. until its ban by the U.S. Food and Drug Administration. Benzalkonium chloride (BAC), a quaternary ammonium compound, has widely replaced triclosan in soaps marketed as an antibacterial. BAC has been detected in surface waters and its presence will likely increase following increased use in soap products. The objective of this study was to determine the effect of BAC on relative abundance of antibiotic resistance in a bacterial community from a surface water used as a source for drinking water treatment. Bench-scale microcosm experiments were conducted with microbial communities amended with BAC at concentrations ranging from 0.1 μg L⁻¹ to 500 μg L⁻¹. Phenotypic antibiotic resistance was quantified by culturing bacteria in the presence of different antibiotics, and genotypic resistance was determined using qPCR to quantify antibiotic resistance genes (ARGs). BAC at concentrations ranging from 0.1 μg L⁻¹ to 500 μg L⁻¹ was found to positively select for bacteria resistant to ciprofloxacin and sulfamethoxazole, and negatively select against bacteria with resistance to six other antibiotics. Exposure to BAC for 14 days increased the relative abundance of sul1 and blaTEM. This study re-highlights the importance of employing both culture and non-culture-based techniques to identify selection for antibiotic resistance. The widespread use of BAC will likely impact antibiotic resistance profiles of bacteria in the environment, including in source waters used for drinking water, wastewater treatment plants, and natural waterways.

Penicillin fermentation dreg (PFD) is a solid waste discharged by pharmaceutical enterprises in the fermentation production process. Due to the residual antibiotic of PFD, the risk of antibiotic resistance bacteria (ARB) generation should be considered in the disposal process. High-throughput quantitative PCR (HT-qPCR) and 16S rRNA gene sequencing were performed to investigate the effect of PFD on the dynamics of antibiotic resistance genes (ARGs) and bacterial community during a lab-scale soil experiment. After the application of PFD, the bacterial number and diversity showed an obvious decrease in the initial days. The abundances of Streptomyces and Bacillus, which are the most widespread predicted source phyla of ARGs, increased remarkably from 4.42% to 2.59%–22.97% and 21.35%. The increase of ARGs was observed during the PFD application and the ARGs carried by PFD itself contributed to the initiation of soil ARGs. The results of redundancy analysis (RDA) show that the shift in bacterial community induced by variation of penicillin content is the primary driver shaping ARGs compositions.

Struvite (MgNH₄PO₃·6H₂O) crystallization is one of important methods of phosphorus recovery from wastewater. As to livestock wastewater, the high-strength occurrence of antibiotics and antibiotic resistance genes might induce struvite recovery to spread antibiotic resistance to the environment. However, limited information has been reported on the simultaneous transport of antibiotics and ARGs in struvite recovery. In the present study, tetracyclines (TCs) and tetracyclines antibiotic resistance genes (ARGs) were selected as the targeted pollutants, and their discrepant residues in struvite recovery from swine wastewater were investigated. TCs and ARGs were obviously detected, with their contents of 4.88–79.5 mg/kg and 6.99 × 10⁷–2.14 × 10¹¹ copies/g, notably higher than those of TCs 0.550–1.94 mg/kg and ARGs 3.98 × 10⁴–5.66 × 10⁷ copies/g obtained from synthetic wastewater. The correlational relationship revealed that predominant factors affecting TCs and ARGs transports were different. Results from network analyses indicated that among the total edges, the negative correlations between TCs and ARGs predominately occupied 18.0%. The redundancy analysis revealed that mineral components in the recovered products, including struvite, K-struvite and amorphous calcium phosphate, coupling with organic contents, displayed insignificant roles on TCs residues, where heavy metals exerted positive and remarkable functions to boost TCs migration. Unexpectedly, mineral components and heavy metals did not displayed significant promotion on ARGs transport as a whole.

Antibiotic resistance is known to impact treatment efficiency of Plesiomonas infections negatively with fatal outcomes. This study investigated antibiogram fingerprint of P. shigelloides (n = 182) isolated from three South Africa rivers using the disc diffusion technique. Environmental pollution and analogous health risk (given infections) that could associate with the freshwaters and empirical treatment of Plesiomonas were assessed using Antibiotic Resistance Index (ARI) and Multiple Antibiotic Resistance Indices (MARI), respectively. Thirteen EUCAST recommended (ERAs) and eleven non-recommended antibiotics (NAs) used as first line agents in the treatment of gastroenteritis and extraintestinal infections were tested. Resistance against ERAs decreased from cefoxitin (37.91%), cefuroxime (35.17%), cefepime (31.87%), ceftriaxone (29.67%), ciprofloxacin (18.13%), trimethoprim-sulfamethoxazole (10.44%), piperacillin/tazobactam (8.79%), ertapenem (4.95%), norfloxacin (4.40%), levofloxacin (2.75%), meropenem (1.10%) to imipenem (0.55%). The isolates had higher resistance (≥36.07%) against NAs but were susceptible to amikacin (67.58%), gentamycin (73.08%), and tetracycline (80.77%). MARI of the isolates were significantly different between ERAs and NAs (P-value < 0.05) and had an average of 0.17 ± 0.18 and 0.45 ± 0.13, respectively. About 33.87% and 95.63% of the isolates had MARI value from 0.23 to 0.62 and 0.27–0.82 to ERAs and NAs, respectively. Also, ERAs-based and NAs-based ARI across sampling units showed significantly different (P-value < 0.05) means of 0.18 ± 0.09 and 0.46 ± 0.05, respectively. MARI attributed low risk of empirical treatment to recommended antibiotics but higher risk to non-recommended antibiotics. Model estimated successful and unsuccessful empirical treatment of infections risks due to resistance in the isolates using recommended antibiotics as 65.93% and 34.07%, respectively; 1.65% and 98.35% in the case of non-recommended antibiotics, respectively. ARI based on recommended antibiotics identified potential environmental pollutions in a number of sites. Resistance in freshwater P. shigelloides especially against cephalosporin, quinolones and fluoroquinolones is distressing and might suggests high pollution of the freshwaters in the Eastern Cape Province.

Mcr-1 and blaNDM₋₁ antibiotic resistance genes (ARGs) confer resistance to colistins and carbapenems, which are often antibiotics used as a last resort in tertiary care hospitals. Dissemination of these two ARGs in drinking water supply systems and their effect on healthy gut bacteria are poorly studied. In this study, the dissemination of mcr-1 and blaNDM₋₁ in a drinking water supply system, and their effect on the antibiotic resistance of mouse gut bacteria are explored.Metagenome analysis revealed that source water (Taipu river and Jinze reservoir) was polluted with ARGs. Mcr-1 and blaNDM₋₁ can be disseminated through the water distribution system. Even advanced water treatments (ozone and biological activated carbon (BAC)) could not effectively remove mcr-1 and blaNDM₋₁. Low concentrations of chloramine disinfectants in the water distribution system were not effective at limiting ARG abundance. Mobile genetic elements were also found to play a major role in the dissemination of ARGs via horizontal gene transfer (HGT) throughout the water supply system. Statistical analysis revealed that there was no effect of temperature on the abundance of mcr-1 and blaNDM₋₁ throughout the water supply system.A last resort ARG, mcr-1 can disseminate from drinking water to the healthy mouse gut. The presence of mcr-1 in a strain belonging to Enterococcus hirae, which is different from the strain belonging to the Bacillus cereus group isolated from drinking water, strongly supports the phenomena of HGT inside the gut.This research provides novel insights into the role of drinking water in disseminating ARGs to the gut and strongly suggests that drinking water may also play a major role apart from other factors known to be involved in the prevalence of last resort ARGs in the gut.

The occurrence of Active Pharmaceutical Ingredients (APIs) in the environment is becoming a major area of concern due to their undesirable effects on non-target organisms. This study investigated the occurrence and risk of contamination by five antibiotics and three antiretrovirals drugs in a fast-growing peri-urban area in Kenya, with inadequate sewer system coverage. Due to poor sewage connectivity and poorly designed decentralized systems, wastewater is directly released in open drains. Water and sediment samples were collected from open surface water drains, while wastewater samples were collected from centralized wastewater treatment plants (WWTP). Solid-phase extraction and ultrasonic-assisted extraction for the aqueous and sediment samples respectively were carried out and extracts analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) using isotopically labeled internal standards. APIs were observed with the detection frequency ranging from 36% to 100%. High mean concentrations of 48.7 μg L⁻¹, 108 μg L⁻¹, and 532 μg L⁻¹ were observed in surface drains for Lamivudine (3 TC), Sulfamethoxazole (SMX), Ciprofloxacin (CIP) respectively. Drain sediments also showed high concentrations of APIs ranging from 2.1 to 13,100 μg kg⁻¹. APIs in this study exceeded those observed in existing literature studies. JKUAT WWTP removal efficiencies varied from −90.68% to 72.67%. Total APIs emission load of the study area was 3550 mg d⁻¹ with WWTP effluent contributing higher loads (2620 mg d⁻¹) than surface water drains (640 mg d⁻¹). Zidovudine (ZDV), nevirapine (NVP), and trimethoprim (TMP) loads in drains, however, exceeded WWTP effluent. Low to high ecotoxicity risk of the individual APIs were observed to the aquatic environment, with high risks for the development of antibiotic resistance in microbiome as determined by the risk quotient (RQ) approach. Risk management through efficient wastewater collection, conveyance, and treatment is necessary to suppress the measured concentrations.