Bile acid profiling as an effective biomarker for staging in pediatric inflammatory bowel disease.

Rapid and accurate clinical staging of pediatric patients with inflammatory bowel disease (IBD) is crucial to determine the appropriate therapeutic approach. This study aimed to identify effective, convenient biomarkers for staging IBD in pediatric patients. We recruited cohorts of pediatric patients with varying severities of IBD to compare the features of the intestinal microbiota and metabolites between the active and remitting disease stages. Metabolites with potential for staging were targeted for further assessment in both patients and colitis model mice. The performance of these markers was determined using machine learning and was validated in a separate patient cohort. Pediatric patients with IBD exhibited distinct gut microbiota structures at different stages of disease activity. The enterotypes of patients with remitting and active disease were Bacteroides-dominant and Escherichia-Shigella-dominant, respectively. The bile secretion pathway showed the most significant differences between the two stages. Fecal and serum bile acid (BA) levels were strongly related to disease activity in both children and mice. The ratio of primary BAs to secondary BAs in serum was developed as a novel comprehensive index, showing excellent diagnostic performance in stratifying IBD activity (0.84 area under the receiver operating characteristic curve in the primary cohort; 77% accuracy in the validation cohort). In conclusion, we report profound insights into the interactions between the gut microbiota and metabolites in pediatric IBD. Serum BAs have potential as biomarkers for classifying disease activity, and may facilitate the personalization of treatment for IBD.

Research Progress of Non-Noble Metal Catalysts for Carbon Dioxide Methanation.

The extensive utilization of fossil fuels has led to a rapid increase in atmospheric CO2 concentration, resulting in various environmental issues. To reduce reliance on fossil fuels and mitigate CO2 emissions, it is important to explore alternative methods of utilizing CO2 and H2 as raw materials to obtain high-value-added chemicals or fuels. One such method is CO2 methanation, which converts CO2 and H2 into methane (CH4), a valuable fuel and raw material for other chemicals. However, CO2 methanation faces challenges in terms of kinetics and thermodynamics. The reaction rate, CO2 conversion, and CH4 yield need to be improved to make the process more efficient. To overcome these challenges, the development of suitable catalysts is essential. Non-noble metal catalysts have gained significant attention due to their high catalytic activity and relatively low cost. In this paper, the thermodynamics and kinetics of the CO2 methanation reaction are discussed. The focus is primarily on reviewing Ni-based, Co-based, and other commonly used catalysts such as Fe-based. The effects of catalyst supports, preparation methods, and promoters on the catalytic performance of the methanation reaction are highlighted. Additionally, the paper summarizes the impact of reaction conditions such as temperature, pressure, space velocity, and H2/CO2 ratio on the catalyst performance. The mechanism of CO2 methanation is also summarized to provide a comprehensive understanding of the process. The objective of this paper is to deepen the understanding of non-noble metal catalysts in CO2 methanation reactions and provide insights for improving catalyst performance. By addressing the limitations of CO2 methanation and exploring the factors influencing catalyst effectiveness, researchers can develop more efficient and cost-effective catalysts for this reaction.

Systematic identification and characterization of five transcription factors mediating the oxidative stress response in Candida albicans.

Candida albicans is an opportunistic human fungal pathogen that causes superficial and systemic infections, particularly in immunocompromised individuals. In response to C. albicans infection, innate immune cells of the host produce and accumulate reactive oxygen species (ROS), which can lead to irreversible damage and apoptosis of fungal cells. Several transcription factors involved in this oxidative stress response have been identified; however, a systematic study to identify the transcription factors that mediate the oxidative stress response has not yet been conducted. Here, we screened a comprehensive transcription factor mutant library consisting of 211 transcription factor deletion mutant strains in the presence and absence of hydrogen peroxide (H2O2), a potent ROS inducer, and identified five transcription factors (Skn7, Dpb4, Cap1, Dal81, and Stp2) that are sensitive to H2O2. Genome-wide transcriptional profiling revealed that H2O2 induces a discrete set of differentially regulated genes among the five identified transcription factor mutant strains. Functional enrichment analysis identified KEGG pathways pertaining to glycolysis/gluconeogenesis, amino sugar and nucleotide sugar metabolism, and ribosome synthesis as the most enriched pathways. GO term analysis of the top common differentially expressed genes among the transcription factor mutant strains identified hexose catabolism and iron transport as the most enriched GO terms upon exposure to H2O2. This study is the first to systematically identify and characterise the transcription factors involved in the response to H2O2. Based on our transcriptional profiling results, we found that exposure to H2O2 modulates several downstream genes involved in fungal virulence. Overall, this study sheds new light on the metabolism, physiological functions, and cellular processes involved in the H2O2-induced oxidative stress response in C. albicans.

The regulatory subunits of CK2 complex mediate DNA damage response and virulence in Candida Glabrata.

BACKGROUND: Candida glabrata which belongs to normal microbiota, has caused significant concern worldwide due to its high prevalence and drug resistance in recent years. C. glabrata has developed many strategies to evade the clearance of the host immune system, thereby causing persistent infection. Although coping with the induced DNA damage is widely acknowledged to be important, the underlying mechanisms remain unclear. RESULTS: The present study provides hitherto undocumented evidence of the importance of the regulatory subunits of CgCK2 (CgCkb1 and CgCkb2) in response to DNA damage. Deletion of CgCKB1 or CgCKB2 enhanced cellular apoptosis and DNA breaks and led to cell cycle delay. In addition, deficiencies in survival upon phagocytosis were observed in Δckb1 and Δckb2 strains. Consistently, disruption of CgCKB1 and CgCKB2 attenuated the virulence of C. glabrata in mouse models of invasive candidiasis. Furthermore, global transcriptional profiling analysis revealed that CgCkb1 and CgCkb2 participate in cell cycle resumption and genomic stability. CONCLUSIONS: Overall, our findings suggest that the response to DNA damage stress is crucial for C. glabrata to survive in macrophages, leading to full virulence in vivo. The significance of this work lies in providing a better understanding of pathogenicity in C. glabrata-related candidiasis and expanding ideas for clinical therapies.

Deciphering the interaction impacts between antiseptic benzethonium chloride and biofilm nitrification system: Performance, resistance mechanisms and biodegradation.

Benzethonium chloride (BEC) is one of emerging bacteriostatic agents. BEC-bearing wastewater generated during sanitary applications in food and medication is easily combined with other wastewater streams to flow into wastewater treatment plants. This study focused on the long-term (231 days) impacts of BEC on the sequencing moving bed biofilm nitrification system. Nitrification performance was tolerant to low concentration of BEC (≤ 0.2 mg/L), but the nitrite oxidation was severely inhibited when the concentration of BEC was 1.0-2.0 mg/L. Partial nitrification maintained about 140 days with nitrite accumulation ratio over 80%, mainly caused by the inhibition of Nitrospira, Nitrotoga and Comammox. Notably, BEC exposure in the system might cause the co-selection of antibiotic resistance genes (ARGs) and disinfectant resistance genes (DRGs), and the resistance of biofilm system to BEC was strengthened by efflux pumps mechanism (qacEdelta1 and qacH) and antibiotic deactivation mechanism (aadA, aac(6')-Ib and blaTEM). Extracellular polymeric substances secretion and BEC biodegradation were also contributed to the system microorganisms resisting BEC exposure. In addition, Klebsiella, Enterobacter, Citrobacter and Pseudomonas were isolated and identified as BEC degrading bacteria. The metabolites of N,N-dimethylbenzylamine, N-benzylmethylamine and benzoic acid were identified, and the biodegradation pathway of BEC was proposed. This study brought new knowledge about the fate of BEC in biological treatment units and laid a foundation for its elimination from wastewater.

Metatranscriptome Revealed the Efficacy and Safety of a Prospective Approach for Agricultural Wastewater Reuse: Achieving Ammonia Retention during Biological Treatment by a Novel Natural Inhibitor Epsilon-Poly-l-Lysine.

Appropriate inhibitors might play important roles in achieving ammonia retention in biological wastewater treatment and its reuse in agriculture. In this study, the feasibility of epsilon-poly-l-lysine (ε-PL) as a novel natural ammonia oxidation inhibitor was investigated. Significant inhibition (ammonia oxidation inhibition rate was up to 96.83%) was achieved by treating the sludge with ε-PL (400 mg/L, 12 h soaking) only once and maintaining for six cycles. Meanwhile, the organic matter and nitrite removal was not affected. This method was effective under the common environmental conditions of biological wastewater treatment. Metatranscriptome uncovered the possible action mechanisms of ε-PL. The ammonia oxidation inhibition was due to the co-decrease of Nitrosomonas abundance, ammonia oxidation genes, and the cellular responses of Nitrosomonas. Thauera and Dechloromonas could adapt to ε-PL by stimulating stress responses, which maintained the organic matter and nitrite removal. Importantly, ε-PL did not cause the enhancement of antibiotic resistance genes and virulent factors. Therefore, ε-PL showed a great potential of ammonia retention, which could be applied in the biological treatment of wastewater for agricultural reuse.

Partial S(0)-driven autotrophic denitrification process facilitated the quick natural enrichment of anammox bacteria at room temperature.

Anaerobic ammonium oxidation (anammox) is well-known to be an environmental and promising biotechnology. However, the natural enrichment of anammox bacteria is still a challenging topic. In this study, partial S(0)-driven autotrophic denitrification (PSAD) was developed to stably supply nitrite, and natural enrichment of anammox bacteria was rapidly realized in a single sequencing moving bed biofilm reactor at room temperature. With the initiation of PSAD, anammox bacteria spontaneously emerged within 12 days, and PSAD-anammox coupling system was realized successfully. And then, the influent concentration of ammonium continuously increased to the same concentration as the nitrate, and the mean total nitrogen removal efficiency reached 92.77 %, which was mainly contributed by anammox. Moreover, the coupling of PSAD and anammox reduced the risk of sulfate emissions. cDNA high throughput sequencing revealed that the relative abundance of Candidatus Brocadia and Thiobacillus reached 39.03 % and 13.48 % at the 88th day. Oligotyping analysis illustrated that GATTTAAT and GTCCCA were the dominant Ca. Brocadia and Thiobacillus oligotypes in PSAD-anammox coupling system, respectively. DNA-based stable isotope probing further deciphered that Thiobacillus was the actual performer of PSAD and supported the nitrite for anammox bacteria in PSAD-anammox coupling system. Overall, this work provided a new strategy to naturally enrich anammox bacteria.

Different acesulfame potassium fate and antibiotic resistance propagation pattern in nitrifying and denitrifying sludge systems.

Acesulfame potassium (ACE-K) is a widely utilized sugar substitute with increasing demand, which is frequently detected in various environmental matrix due to recalcitrance. However, a general consensus on the contribution of nitrifying and denitrifying process to ACE-K removal is lacking. Therefore, ACE-K removal, its effects on antibiotic resistant genes (ARGs) propagation and microbial community in nitrifying sequencing batch reactor (N-SBR) and denitrifying sequencing batch reactor (D-SBR) inoculated with the identical activated sludge were investigated. In this study, ACE-K can be eliminated in N-SBR with satisfying removal efficiency (96.76 ± 8.33 %) after 13 d acclimation, while it remained persistent (average ACE-K removal efficiency of 2.24 ± 1.86 %) in D-SBR during 84 d exposure. Moreover, ACE-K hardly affected the performances of these two types of reactors and had little impact on nitrifying and denitrifying functional genes. However, initial contact with ACE-K would increase ARGs abundance, network analysis showed functional bacteria in each reactor were possible ARGs hosts. Potential ACE-K degrading genera Chelatococcus, Bosea and Aquamicrobium were found in both reactors. LefSe analysis showed that Phyllobacteriaceae containing Aquamicrobium genus was a differentially enriched family in N-SBR. This research might provide a perspective for better understanding factor affecting ACE-K fate in wastewater treatment process and its ecological risks.

Quaternary ammonium compounds promoted anoxic sludge granulation and altered propagation risk of intracellular and extracellular antibiotic resistance genes.

Surfactants could influence sludge morphology and disinfectants were linked to antibiotic resistance genes (ARGs). Thus, the response of activated sludge and ARGs to long-term quaternary ammonium compounds (QACs) exposure required further investigation, which is a popular surfactant and disinfectant. Here, three sequencing batch reactors were fed with 5 mg/L most frequently detected QACs (dodecyl trimethyl ammonium chloride (ATMAC C12), dodecyl benzyl dimethyl ammonium chloride (BAC C12) and didodecyl dimethyl ammonium chloride (DADMAC C12)) for 180 d. The long-term inhibitory effect on denitrification ranked: DADMAC C12 > BAC C12 > ATMAC C12. Besides, obvious granular sludge promoted by the increase of α-Helix/(ß-Sheet + Random coil) appeared in DADMAC C12 system. Moreover, intracellular ARGs increased when denitrification systems encountered QACs acutely but decreased in systems chronically exposed to QACs. Although replication and repair metabolism in ATMAC C12 system was higher, ATMAC C12 significantly promoted proliferation of extracellular ARGs. It was noteworthy that the propagation risk of extracellular ARGs in sludge increased significantly during sludge granulation process, and intracellular sul2 genes in sludge and water both increased with the granular diameter in DADMAC C12 system. The universal utilization of QACs may enhance antibiotic resistance of bacteria in wastewater treatment plants, deserving more attention.

Unraveling the impact and mechanism of antipyretic paracetamol on intergenera conjugative plasmid transfer.

Antimicrobial resistance has been considered as a great threat to biosecurity and human health. And the transmission of antibiotic resistance genes (ARGs) by conjugated plasmid is a key factor in the prevalence of antimicrobial resistance. Paracetamol (PRC), one of nonopioid analgesics, is an extensively used antipyretic and mild analgesic worldwide available for numerous prescriptions. It was unclear whether PRC could promote the spread of ARGs. Here, it was demonstrated that PRC promoted intergenera conjugative plasmid transfer in an established conjugation model. Both donor and recipient strains treated by PRC emerged the variations of reactive oxygen species (ROS), SOS response and cell membrane permeability. Correspondingly, transcriptome analysis revealed that the gene expression involved in cell membrane permeability and SOS response was up-regulated significantly after PRC exposure. More directly, PRC also increased the expressions of conjugation related genes of trbG and trbP in donor. This study proved for the first time that PRC could enhance the intergenera conjugative plasmid transfer. Collectively, these findings manifested the potential threat associated with the existence of non-antibiotic substance PRC, which could provide an important insight into antimicrobial resistance spread.

The fate and behavior mechanism of antibiotic resistance genes and microbial communities in flocs, aerobic granular and biofilm sludge under chloroxylenol pressure.

Chloroxylenol (PCMX), an antibacterial agent, has been widely detected in water environment and has toxic effects on biology and ecology. During 270 d, the influence of PCMX on the performance of three nitrification systems was investigated, including floc-based sequencing batch reactor (FSBR), aerobic granule-based SBR (AGSBR) and biofilm SBR (BSBR). The nitrification capability of three systems was inhibited by PCMX, but recovered after domestication, and PCMX made three systems realize partial nitrification for 10, 100 and 35 days, respectively. The extracellular polymeric substances of three systems increased first and then decreased with the increment of PCMX. The granular structure of AGSBR may be conducive to the enrichment of antibiotic resistance genes (ARGs), and almost all ARGs of BSBR were reduced during the addition of 5.0 mg/L PCMX. The microbial community results showed that Rhodococcus as potential degrading bacteria was continuously enriched in three systems. Piscinibacter was regarded as the potential antibiotic resistant bacteria, which was positively associated with multiple ARGs in three systems. Additionally, quaternary ammonium compounds resistance genes had a variety of positive correlations with bacteria in three systems. This study provided a new perspective for the usage and treatment of PCMX.

Distinct bacterial communities and resistance genes enriched by triclocarban-contaminated polyethylene microplastics in antibiotics and heavy metals polluted sewage environment.

Knowledge gaps still surround the question of what biofilms form on contaminated microplastics (MPs) in the antibiotics and (or) heavy metals polluted sewage. In this work, the clean polyethylene microplastics (PE MPs) and triclocarban (TCC)-contaminated PE MPs were cultured in the sewage containing only ampicillin (AMP), only copper (Cu) and both AMP and Cu for 28 days. The results showed that the TCC on PE MPs (with concentration of 2.48 mg/g PE MPs) did not impede the adhesion of the bacteria and the formation of biofilm. Moreover, many potential pathogenic bacteria (Aquabacterium and Pseudoxanthomonas) and potential resistant bacteria (Stenotrophomonas) were more likely to attach on TCC-contaminated PE MPs compared with clean PE MPs. In addition, biofilms of TCC-contaminated PE MPs had highest potential pathogenic functions. TCC-contaminated PE MPs also caused the increases of various resistance genes in both biofilm and sewage. The co-occurrence of TCC, AMP and Cu might exert a stronger selective pressure on bacterial communities and promote the co-selection of resistance genes. In addition, TCC-contaminated PE MPs resulted in higher abundance of five mobile genetic elements (MGEs) (intI1, intI3, tnpA-04, IS613 and trb-C) in sewage, which might further promote the transmission of resistance genes.

Higher spreading risk of antibacterial biocide and heavy metal resistance genes than antibiotic resistance genes in aerobic granular sludge.

Metagenomic approach was applied to simultaneously reveal the antibiotic resistance genes (ARGs) and antibacterial biocide & metal resistance genes (BMRGs), and the corresponding microbial hosts with high mobility during aerobic granular sludge (AGS) formation process. The results showed that the relative abundance of BMRGs was 88-123 times that of ARGs. AGS process was easier to enrich BMRGs, leading to a greater risk of drug resistance caused by BMRGs than that by ARGs. The enrichments of ARGs and BMRGs in AGS were closely related to several enhanced microbial metabolisms (i.e., cell motility, transposase and ATP-binding cassette transporters) and their corresponding regulatory genes. Several enhanced KEGG Orthologs (KO) functions, such as K01995, K01996, K01997 and K02002, might cause a positive impact on the spread of ARGs and BMRGs, and the main contributors were the largely enriched glycogens accumulating organisms. The first dominant ARGs (adeF) was carried by lots of microbial hosts, which might be enriched and propagated mainly through horizontal gene transfer. Candidatus Competibacter denitrificans simultaneously harbored ARG (cmx) and Cu related RGs (corR). Many enriched bacteria contained simultaneously multiple BMRGs (copR and corR) and mobile genetic elements (integrons and plasmids), granting them high mobility capabilities and contributing to the spread of BMRGs. This study might provide deeper understandings of the proliferation and mobility of ARGs and BMRGs, importantly, highlighted the status of BMRGs, which laid the foundation for the controlling widespread of resistance genes in AGS.

Feeding low-level benzethonium chloride can promote the start-up, fast recovery and long-term stable maintenance of partial nitrification for low-ammonium wastewater.

The establishment of stable partial nitrification (PN) is beneficial to promote the application of anaerobic ammonium oxidation, especially for low-ammonium wastewater. This study demonstrated an innovative approach for achieving PN through feeding low-level benzethonium chloride (BZC). PN was started and maintained for 125 days after the sequential feeding of 0.2 and 1 mg/L BZC for 50 days. The damaged PN recovered rapidly within eight days by feeding 2 mg/L BZC, and it thrived for more than 172 days, indicating that nitrite-oxidizing bacteria did not adapt to BZC. The removal of BZC changed from adsorption to degradation gradually. Increased extracellular polymeric substances secretion and altered protein secondary structures explained sludge granulation during BZC feeding, which may be closely related to long-term stable maintenance of PN. PICRUSt2 revealed the underlying microbial mechanisms in depth. Overall, this research proposed a novel scheme to achieve robust PN treating low-ammonium wastewater through feeding low-level BZC.

Fates of quaternary ammonium compound resistance genes and the corresponding resistant strain in partial nitrification/anammox system under pressure of hexadecyl trimethyl ammonium chloride.

Hexadecyl trimethyl ammonium chloride (ATMAC-C16) is a kind of quaternary ammonium compound (QACs) which is extensively consumed as disinfectants, antimicrobials and surfactants. Here, the partial nitrification/anammox (PN/A) system was exposed to different levels of ATMAC-C16 (0-10 mg/L) and the main objective was to reveal the long-term microbiological responses of PN/A system to ATMAC-C16, importantly, explore the tolerance of PN/A to ATMAC-C16 and the key resistant strain. Nitrogen removal efficiency was influenced by environmental and extreme levels of ATMAC-C16 through mainly affecting the anammox (hzsB) gene. Two types of anammox, Candidatus Jettenia and Candidatus Kuenenia, were enriched under the pressure of ATMAC-C16, which allowed PN/A system to maintain good nitrogen removal performance. ATMAC-C16 might cause the hormesis of entire microbial population in PN/A system, leading to the enhancement of cell viability. ATMAC-C16 decreased the relative abundances of most antibiotics resistance genes (ARGs) but significantly enriched QACs resistance genes (QRGs). The tolerance of PN/A system to ATMAC-C16 might be strengthened by inducing the efflux pumps encoding genes (qacH-01/02). Microbial hosts dynamic and co-selection mechanism among ARGs and QRGs resulted in the opposite trends of qacEdeltal-01/02 and qacH-01/02. Pseudoxanthomonas mexicana was identified as the ATMAC-C16 resistant strain, and its resistance to 10 mg/L ATMAC-C16 might not only obtain by capturing the qacH gene, but also benefit from its own efflux pump system. Therefore, from the perspective of the transmission of resistance genes, especially for QRGs, the spread risk of QRGs and ATMAC-C16 resistant strain in PN/A technique should be taken seriously.

Triclosan enriched resistance genes more easily than copper in the presence of environmental tetracycline in aerobic granular sludge system.

Triclosan (TCS) and copper (Cu2+) were exposed to aerobic granular sludge (AGS) system treating wastewater containing environmental tetracycline, respectively, to explore the different biochemical responses, more importantly, the fates of resistance genes (RGs) in AGS system. The results showed that TCS and Cu2+ could significantly inhibit the N and P removal in AGS system by reducing several key functional genes, including amoA gene of ammonia-oxidizing bacteria, Nitrospira and phosphorus accumulating organisms 16S rRNA genes. TCS caused higher degree of RGs' enrichment than Cu2+, which made the average total relative abundance of RGs of 1.38 ± 0.73 and 0.78 ± 0.24 in TCS and Cu system, respectively. Cu2+ could induce a wider range of horizontal gene transfer than TCS, leading to the detections of more potential hosts harboring RGs in Cu system. Cu system seemed to have stronger repair, immunity and defense ability than TCS system, which enabled it to have sufficient ability to trigger protection mechanism to realize self-protection, eventually the RGs also were controlled. Integron (intI1 and intI3) and plasmids (trb-C and IncQ) might cooperate with microorganisms and water quality parameters to enhance the enrichment of RGs in TCS system, however this interaction among various environmental factors was not obvious in Cu system, which might be responsible for the lower abundance of RGs. The increasing levels of TCS and Cu2+ in wastewater should be paid more attentions during the treatment of wastewater containing environmental tetracycline by AGS system. Especially for TCS, it had the ability to enrich RGs more easily than Cu2+, which should be prevented from entering wastewater treatment plants as far as possible, to avoid more serious proliferation and dissemination of various RGs.

Robustness of the partial nitrification-anammox system exposing to triclosan wastewater: Stress relieved by extracellular polymeric substances and resistance genes.

The partial nitrification-anammox (PN/A) process is a promising method for the treatment of municipal wastewater. It is necessary to clarify the responses of PN/A system to antimicrobial agent triclosan (TCS) widely existed in the influent of wastewater treatment plants. In this study, it was found that PN/A system was robust to cope with 0.5 mg/L TCS. Specifically, the control reactor reached 80% total nitrogen removal efficiency (TNRE) on day 107, while the reactor feeding with 0.5 mg/L TCS reached the same TNRE on day 84. The results of the activity test, high-throughput sequencing and DNA-based stable isotope probing showed that 0.5 mg/L TCS did not impede the performance of ammonia oxidizing archaea, ammonia oxidizing bacteria (Nitrosomonas) and anammox bacteria (Candidatus Brocadia and Ca. Kuenenia), but significant inhibited the nitrite oxidizing bacteria (Nitrospira and Ca. Nitrotoga) and denitrifying bacteria. The influent TCS led to the increase of EPS content and enrichment of four resistance genes (RGs) (intI1, sul1, mexB, and tnpA), which might be two principal mechanisms by which PN/A can resist TCS. In addition, functional bacteria carrying multiple RGs also contributed to the maintenance of PN/A system function. These findings improved the understandings of antimicrobial effects on the PN/A system.

Chloroxylenol at environmental concentrations can promote conjugative transfer of antibiotic resistance genes by multiple mechanisms.

The intergeneric conjugative transfer of antibiotic resistance genes (ARGs) is recognized as an important way to the dissemination of antibiotic resistance. However, it is unknown whether the extensive use of chloroxylenol (para-chloro-meta-xylenol, PCMX) in many pharmaceutical personal care products will lead to the spread of ARGs. In this study, the abilities and mechanisms of PCMX to accelerate the intergeneric conjugative transfer were investigated. Results showed that exposure of bacteria to environmental concentrations of PCMX (0.20-1.00 mg/L) can significantly stimulate the increase of conjugative transfer by 8.45-9.51 fold. The phenotypic experiments and genome-wide RNA sequencing revealed that 0.02-5.00 mg/L PCMX exposure could increase the content of alkaline phosphatase and malondialdehyde, which are characteristic products of cell wall and membrane damage. In addition, PCMX could lead to excessive production of reactive oxygen species (ROS) by 1.26-2.00 times, the superoxide dismutase and catalase produced by bacteria in response to oxidative stress were not enough to neutralize the damage of ROS, thus promoting the conjugative transfer. Gene Ontology enrichment analysis indicated that cell membrane permeability, pili, some chemical compounds transport and energy metabolism affected conjugative transfer. This study deepened the understanding of PCMX in promoting propagation of ARGs, and provided new perspectives for use and treatment of personal care products.

Enhancement of antibiotic resistance dissemination by artificial sweetener acesulfame potassium: Insights from cell membrane, enzyme, energy supply and transcriptomics.

The abuse of antibiotics on animals could induce the development of antibiotic resistant genes (ARGs) and antibiotic resistant bacteria (ARB), and acesulfame potassium (ACE) is the widely used artificial sweetener in animal feed. Generally speaking, ACE and ARB often coexist in livestock wastewater, however, the impact of the co-occurrence of ACE and ARB on the transmission of ARGs is still unknown. In this study, the effects of ACE on vertical gene transfer (VGT) and horizontal gene transfer (HGT) were both evaluated. For VGT, ACE may hinder the spread of sul gene in Pseudomonas HLS-6 by blocking ARB growth. As for HGT (from Escherichia coli DH5α to Pseudomonas HLS-6), environmentally relevant ACE concentration could facilitate the conjugative transfer. The underlying mechanisms of HGT were characterized by enhanced cell membrane permeability, reactive oxygen species overproduction, SOS response, energy supply, which were all further verified by the changes in transcription levels of related genes. Interestingly, intracellular Mg2+ in donor strain was found for the first time as an indicator for the conjugation occurrence in ACE treated mating system. This study may provide new insights into the role of ACE on ARGs proliferation and highlight its potential environmental impacts.

Triclocarban shifted the microbial communities and promoted the spread of antibiotic resistance genes in nitrifying granular sludge system.

Triclocarban (TCC) is in great market demand especially after the outbreak of COVID-19 pandemic, becoming an emerging pollutant. However, the impacts of TCC on the performance of nitrifying granular sludge system and the occurrence of antibiotic resistance genes (ARGs) were still unknown. This work explored the impacts of different concentrations of TCC on nitrifying granular sludge. Results showed that TCC suppressed the activities of ammonia-oxidizing microorganisms and decreased the abundance of Nitrospira. Adsorption was the main way for the removal of TCC and the biodegradation efficiency of TCC increased to 28.00% under 19.70 mg/L TCC addition. TCC enriched the ARGs and promoted the risks of their transferring in microorganisms. Pseudomonas might not only have strong resistance to TCC, but also propagate ARGs. The removal process of TCC and bacterial communities were important factors to promote the spread of ARGs. Thus, the existence of TCC presented a great environmental risk.