Antibiotic Selection Pressure Research Articles

ESBL-producing strains isolated from imported cases of enteric fever in England and Wales reveal multiple chromosomal integrations of blaCTX-M-15 in XDR Salmonella Typhi.

There are approximately 300 cases of enteric fever reported annually from England and Wales; most are imported infections. Clinical management of enteric fever remains a challenge with the emergence of ESBL-producing strains, especially XDR Salmonella Typhi from Sindh, Pakistan. All strains of S. Typhi and Salmonella Paratyphi A isolated from cases presenting with symptoms of enteric fever in England and Wales, between 1 April 2014 and 31 March 2020, were characterized using WGS. Antibiotic susceptibility testing was performed using an agar dilution method. ESBL strains contributed to 69 cases of enteric fever (S. Typhi n = 68, S. Paratyphi A n = 1); 68 were imported (Pakistan n = 64, Iraq n = 2, Bangladesh n = 1 and India n = 1). Ages ranged from 1 to 56 years, 36/69 (52%) were children, 52% were female and the duration of hospital stay ranged from 1 to 23 days. The ESBL phenotype was conferred by the presence of blaCTX-M-15 (S. Typhi n = 67 and S. Paratyphi A n = 1) or blaCTX-M-55 (S. Typhi n = 1). An IncY plasmid harbouring blaCTX-M-15 and qnr was detected in 56 strains from Pakistan. The IncY plasmid was absent in the remaining strains and there was evidence of a 4 kb ISEcpl-blaCTX-M-15-tnp gene cassette insertion into the chromosome at one of three integration points. Chromosomal integration of blaCTX-M-15 within the XDR Sindh strains may lead to the maintenance of resistance in the absence of antibiotic selection pressure. Empirical treatment of cases of complicated enteric fever returning from Pakistan will henceforth have to include a carbapenem.

Molecular characterization of two novel NDM-1-producing atypical enteroaggregative Escherichia coli isolates from patients

To investigate NDM-1-producing atypical Enteroaggregative Escherichia coli (aEAEC) of sequence type 349 from hospitalized patients, the isolates 13ZX28 and 13ZX36 were subjected to antimicrobial susceptibility testing, conjugation and whole genome sequencing. Only one single nucleotide mutation was detected in chromosomes despite different plasmid profiles. Both isolates were positive for blaNDM-1 mediating resistance to carbapenem. A novel plasmid p13ZX28–272 (~272-kb) from 13ZX28 encodes blaNDM-1. Interestingly, its sequence was identical to the two plasmids p13ZX36–200 (~200-kb) and p13ZX36–70 (~70-kb) from 13ZX36. Formation of the former episome possibly involved homologous recombination through a 4948-bp large fragment located on each of the two latter plasmids. Furthermore, plasmid p13ZX28–272 could be resolved into a ~ 98-kb daughter plasmid by IS26 rearrangement following conjugation. The plasticity of the plasmids is recognized, which warrants further investigation to evaluate the underlying public health risk and understand how antibiotic selection pressure drives this process.

An alternative index for evaluating AMU and anti-methicillin-resistant Staphylococcus aureus agent use: A study based on the National Database of Health Insurance Claims and Specific Health Checkups data of Japan

IntroductionAnti-methicillin-resistant Staphylococcus aureus (MRSA) agents have different doses and administration periods. Thus, it is difficult to evaluate antimicrobial use (AMU) of anti-MRSA agents using defined daily doses per 1000 inhabitants per day (DID) or days of therapy per 1000 inhabitants per day (DOTID). This study aimed to evaluate the relationship between anti-MRSA agent use and resistant bacteria using the number of patients per 1000 inhabitants per day (PID) as an alternative index of AMU. MethodsAMU data for anti-MRSA agents were collected from the National Database of Health Insurance Claims and Specific Health Checkups (NDB) in 2016. The relationship between PID and DID or DOTID was evaluated. The number of patients with MRSA isolated was obtained from Japan Nosocomial Infections Surveillance, and their correlation with PID was analyzed. The rate of anti-MRSA agent use in each prefecture was investigated. ResultsPID showed a significant linear relationship with both DID and DOTID (all p < 0.0001). PID was significantly correlated with the number of patients with MRSA isolated. Additionally, the rate of anti-MRSA agent use was markedly different in each region. ConclusionsPID is not affected by doses and administration periods, and thus may be an alternative index for the selective pressure of antibiotics. Evaluating AMU using PID based on NDB data will help in the development of effective antimicrobial resistance measures.

Antimicrobial Sensitivity Profile Of Uropathogens In A Macapá Laboratory, Amapá, Brazilian Amazon

Urinary tract infections (UTI) represent a worldwide health problem. Microbial resistance, due to selective antibiotic pressure, has a direct influence on the evolution and impact of these infections. The objective of this work was to identify the antimicrobial susceptibility profile of uropathogens isolated in uroculture samples from a private laboratory in the city of Macapá/AP. It is a quantitative, retrospective and cross-sectional study, using a laboratory database. Data were collected from results of urocultures with antibiogram analyzed from January to December 2019. The present study evaluated 3,510 urocultures, carried out during the study period, of which 1,269 had bacterial growth, equivalent to 36.15%. Among the positive results, we found the bacteria Escherichia coli (66.59%), Staphylococcus aureus (32.62%), mixed colonies of E. coli and S. aureus (0.47%), Staphylococcus saprophyticus (0.24%) and Serratia marcescens (0.08%). The male gender was responsible for 16.35% (n = 574) of the urocultures analyzed, while the female population was 83.65% (n = 2936). When analyzing the distribution of bacteria isolated by gender, males were considered a protective factor with 42% less chance of presenting bacteria in urine. The bacteria E. coli was the predominant pathogen in these infections in both sexes and in all age groups.

Insights into enhanced biodegradation of sulfadimethoxine by catalyst: Transcriptomic responses and free radical interactions

The occurrence of sulfonamides in the environment is a severe global threat to public health due to the increasing prevalence of antibiotic selection pressure that may lead to the development of antibiotic resistance. We report an enhanced biodegradation of sulfadimethoxine (SDM) by Phanerochaete chrysosporium (Pc) with lignocellulosic biomass (Lb) using Fe3O4-ZSM-5 as a catalyst (Pc/Fe3O4-ZSM-5/Lb). SDM was completely degraded within 4 days at pH 7.0 in the Pc/Fe3O4-ZSM-5/Lb system. Transcriptomic, metabolites and free radical analyses were performed to explore the detailed molecular mechanisms of SDM degradation. A total of 246 genes of Pc in the Pc/Fe3O4-ZSM-5/Lb system exhibited significant upregulation compared to that in Pc alone. Upregulated genes encoding cellulases, cytochrome P450, cellobiose quinone oxidoreductase, and cellobiose dehydrogenase were involved in SDM degradation in the Pc/Fe3O4-ZSM-5/Lb system. In addition, genes encoding glutathione S-transferase and cytochrome P450 genes related to oxidative stress and detoxification were all significantly upregulated (P < 0.01). Electron paramagnetic resonance revealed the generation of OH suggesting a free radical pathway could be catalyzed by Fe3O4-ZSM-5 and the enzymes. These findings of catalyst-assisted SDM biodegradation will be valuable for remediation of antibiotics from contaminated wastewater.

Transferable blaCTX-M Carrying Multidrug Resistant Escherichia coli from Pig Population of North Eastern Region of India

Background: We investigated the occurrence of blaCTX-M carrying extended spectrum beta lactamase (ESBL) producing Escherichia coli in pigs from 8 North-eastern states of India with special emphasis on the transferability of ESBL gene from resistant E. coli strains to the susceptible Salmonella strains by in vitro and in vivo.Methods: Fecal samples (n=790) were collected from pigs reared under organized and unorganized farming set up of entire North-eastern region of India. All the samples were processed for isolation and identification of E. coli. All the isolates were subjected to antimicrobial sensitivity assay by disc diffusion method followed by determination of ESBLs producing ability by double disc synergy test (DDST). All the ESBLs producing isolates were screened for blaCTX-M gene by PCR using specific primers. The representative blaCTX-M gene positive isolates were used as donor to determine the ability to transfer of resistance gene in Salmonella by in vitro and in vivo assays with and without antibiotic selection pressure. Result: A total of 2,291 E. coli was isolated, of which 1113 and 1178 were from organized and unorganized farms, respectively. Majority of the isolates were multi-drug resistant with highest resistance against amoxicillin (84.81%) followed by cefalexin (77.17%), sulphafurazole (56.79%), piperacillin (46.40%), tetracycline (38.29%) and cefexime (35.66%). Isolates from unorganized farms showed higher resistance than the isolates recovered from organized farms. A total of 654 (28.55%) isolates were confirmed as ESBL producers by double disc synergy test (DDST) method, of which 65 (2.84%) isolates were positive for blaCTX-M gene. Genotypically, isolates with specific amino acids substitution revealed variation in their antibiotic susceptibility by phenotypic method. blaCTX-M gene could be successfully transferred horizontally from E. coli (donor) to Salmonella (recipient) by in vitro (3.6±2.07x10-8 to 4.4±2.88x10-8 transconjugate per donor) and in vivo method. By in vivo method in pig model, the frequency of transfer was higher under the antibiotic selection pressure (6.6±3.05x10-5 to 7.2±1.92x10-5 trans-conjugants per donor) than without antibiotic pressure (5.6±2.3x10-4 to 6.8±3.35x10-4 trans-conjugants per donor).

A framework for identifying the recent origins of mobile antibiotic resistance genes

Since the introduction of antibiotics as therapeutic agents, many bacterial pathogens have developed resistance to antibiotics. Mobile resistance genes, acquired through horizontal gene transfer, play an important role in this process. Understanding from which bacterial taxa these genes were mobilized, and whether their origin taxa share common traits, is critical for predicting which environments and conditions contribute to the emergence of novel resistance genes. This knowledge may prove valuable for limiting or delaying future transfer of novel resistance genes into pathogens. The literature on the origins of mobile resistance genes is scattered and based on evidence of variable quality. Here, we summarize, amend and scrutinize the evidence for 37 proposed origins of mobile resistance genes. Using state-of-the-art genomic analyses, we supplement and evaluate the evidence based on well-defined criteria. Nineteen percent of reported origins did not fulfill the criteria to confidently assign the respective origin. Of the curated origin taxa, >90% have been associated with infection in humans or domestic animals, some taxa being the origin of several different resistance genes. The clinical emergence of these resistance genes appears to be a consequence of antibiotic selection pressure on taxa that are permanently or transiently associated with the human/domestic animal microbiome.

Microdiversity of Enterococcus faecalis isolates in cases of infective endocarditis: selection of non-synonymous mutations and large deletions is associated with phenotypic modifications

ABSTRACT Context: Today, infective endocarditis (IE) caused by Enterococcus faecalis represents 10% of all IE and is marked by its difficult management and the frequency of relapses. Although the precise reasons for that remain to be elucidated, the evolution of the culprit strain under selective pressure through microdiversification could be, at least in part, involved. Material and methods: To further study the in situ genetic microdiversity and its possible phenotypic manifestations in E. faecalis IE, we sequenced and compared multiple isolates from the valves, blood culture and joint fluid of five patients who underwent valvular surgery. Growth rate and early biofilm production of selected isolates were also compared. Results: By sequencing a total of 58 E. faecalis genomes, we detected a considerable genomic microdiversity, not only among strains from different anatomical origins, but also between isolates from the same studied cardiac valves. Interestingly, deletions of thousands of bases including the well-known virulence factors ebpA/B/C, and srtC, as well as other large prophage sequences containing genes coding for proteins implicated in platelet binding (PlbA and PlbB) were evidenced. The study of mutations helped unveil common patterns in genes related to the cell cycle as well as central metabolism, suggesting an evolutionary convergence in these isolates. As expected, such modifications were associated with a significant impact on the in-vitro phenotypic heterogeneity, growth, and early biofilm production. Conclusion: Genome modifications associated with phenotypic variations may allow bacterial adaptation to both antibiotic and immune selective pressures, and thus promote relapses.

182. increasing Odds of Resistance for Subsequent Urinary e. Coli Isolates

BackgroundAnnual cumulative antibiograms are routinely used by clinicians to guide selection of empirical antibiotic therapies. CLSI guidelines recommend that these antibiograms to analyze data yearly, include only final, verified results, include bacterial species with > 30 isolates and to include only the first isolate for each species/patient instance per analysis period. Handling multiple isolates from individual patients in cumulative antibiograms is a controversial topic within the antimicrobial stewardship community. Current practice favors removing subsequent isolates, thereby discarding data reflecting impact of selective antibiotic pressure on resistance patterns in recurring urinary tract infection (UTI).In this study we analyzed a five-year data set of deidentified outpatient antibiotic results from a commercial laboratory to determine whether there were significant differences in resistance patterns between first and subsequent isolates from the same patient.MethodsThe 5-year antibiotic susceptibility data was restricted to urinary Escherichia coli (EC) isolates. Patient occurrence(s) of urinary EC were categorized by frequency: 1st occurrence, 2nd occurrence, 3rd occurrence, and 4th or greater occurrence. A logistic regression analysis using a binary outcome for resistance and independent variable of patient isolate occurrence was run for amoxicillin-clavulanate, ampicillin, ceftriaxone, ciprofloxacin, gentamicin, levofloxacin, nitrofurantoin, and trimethoprim-sulfa.ResultsFrom a logistic regression analysis, we estimate that for each occurrence in the data, an isolate’s odds of resistance were higher for every increase in a patient’s number of occurrences in the data for all antibiotics reported with p values < 0.0001.Table 1: Odds ratios (OR) of resistance for each subsequent urinary EC isolate occurrence over 5 years ConclusionOur findings suggest that individuals with higher numbers of urinary EC occurrences have more resistant EC than the first EC occurrence, with effects that vary by antibiotic class. Although traditional antibiograms include only the first occurrence of urinary EC from a single patient, this approach may underestimate levels of reservoir resistance in a community. Such an underestimation likely impacts efficacy of empiric therapeutic choice, healthcare outcomes, and cost.Disclosures All Authors: No reported disclosures

201. Healthcare utilization outcomes of patients prescribed fluoroquinolones on discharge from the hospital to nursing homes

BackgroundFluoroquinolones (FQs) are frequently prescribed in nursing homes (NHs) despite concerns regarding broad spectrum antibiotic selective pressure, increased risk of Clostridioides difficile infection, and other adverse events. NH antibiotics are also frequently initiated in hospitals prior to NH admission. We quantified the frequency and outcomes of patients prescribed FQs on discharge from the hospital to NHs.MethodsThis was a retrospective cohort study of adult (age ≥ 18 years) inpatients prescribed a FQ on discharge from Oregon Health & Science University Hospital (OHSU) to a NH between 1/1/2016 and 12/31/2018. Study data were collected from a repository of electronic health record data. The outcome of interest was a composite of 30-day hospital readmission or emergency department (ED) visit to OHSU. Associations were quantified using odds ratios (ORs) and 95% confidence intervals (CIs).ResultsAmong 9,546 patients discharged to a NH, 2,410 (25%) were prescribed at least one antibiotic and 423 (17.6%) were prescribed a FQ. Of these patients, 36.9% were age ≤ 65, 53% were male, 11.6% received a specialty infectious diseases consultation, 34.8% had a surgical diagnosis, and 49.7% had a hospital length of stay > 7 days. The most prevalent comorbidities were cancer (30.5%), chronic obstructive pulmonary disease (29.6%), and renal disease (26%). The most prevalent FQs prescribed were ciprofloxacin (56.7%), levofloxacin (40.2%), and moxifloxacin (3.1%). Duration of NH therapy > 7 days occurred in 37.6% of patients. The most common infectious diagnoses were bloodstream infection and endocarditis (39%), pneumonia (17%), and urinary tract infection (14.2%). Of patients prescribed a FQ, 276 (65.3%) had an ED visit or hospital admission to index facility within 30 days of discharge. Patients who were ≤ 65 years old (OR 2.3, 95% CI 1.4–3.5), male (OR 1.6, 95% CI 1.1–2.5), had comorbid renal disease (OR 1.8, 95% CI 1.1–2.9), or osteomyelitis as infectious diagnosis (OR 2.4, 95% CI 1.0–5.7) were more likely to have a 30-day ED visit or hospital admission.ConclusionPatients prescribed FQs on discharge to NHs frequently returned to the hospital for an ED visit or inpatient admission within 30 days of discharge.Disclosures All Authors: No reported disclosures

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.

TonB-Dependent Receptors Affect the Spontaneous Oxytetracycline Resistance Evolution in Aeromonas hydrophila.

It is well known that most microbial populations develop their intrinsic antibiotics resistance at low concentrations in antibiotics environments, but the factors influencing spontaneous resistance are still largely unknown. In this study, Aeromonas hydrophila strains with different resistance levels to oxytetracycline (OXY) were induced by sublethal antibiotic selection pressure, and differential expression of proteins was compared among them using iTRAQ-based quantitative proteomics. Our following bioinformatic analysis showed that energy metabolism-related proteins were downregulated, while several iron-related proteins were upregulated in high OXY-resistant strains. To further investigate the role of spontaneous OXY resistance evolution, four TonB-dependent receptor-coded genes were deleted, and their OXY susceptibility capabilities and antibiotic evolutionary assays were performed, respectively. Our results showed that the deletion of these genes did not affect the susceptibility to OXY, but showed different evolution rates in the spontaneous OXY evolution compared with wild-type strain, especially for AHA_0971 and AHA_4251. Therefore, this study indicates the important role of TonB-dependent receptor proteins during the bacterial antibiotics resistance evolution and may provide a new prophylactic strategy against the development of antibiotic resistance.

Expansion of a Subset Within the C2 Subclade of Escherichia coli Sequence Type 131 (ST131) Is Driving the Increasing Rates of Aminoglycoside Resistance

BackgroundSequence type 131 (ST131) of Escherichia coli is a pandemic clone that drives the increasing rates of antibiotic resistance. While the pervasiveness of ST131 clade C, especially subclades C2 and C1-M27, has been demonstrated in numerous global surveys, no report about the ST131 clades and their virotypes has been published from Iran so far.MethodsA collection of 73 consecutive ST131 isolates from extraintestinal specimens was investigated for determination of virotypes, antibiotic susceptibility patterns, resistance/virulence determinants, and clade subsets.ResultsMost of the isolates belonged to subclade C2 (33/73; 45.2%), which had the highest virulence factor (VF) scores and resistance rates, followed by C1-M27 (18; 24.6%), C1-non-M27 (14; 19.1%), and A (8; 10.9%). The distinctive profiles of subclade C2 virulence genes were revealed by principle coordinates analysis testing. The distribution of the hlyA virulence gene among subclade C2 was not uniform, so that positive strains (21; 63.6%) showed significantly higher rates of resistance (blaCTX-M-15, blaOXA-1, aac(6’)-Ib-cr, aac(6’)-Ib, aac(3)-IIa) and virulence (hra, tia/hek, K5, cnf, papGII, papC) markers and gentamicin/tobramycin resistance. Virotype C as the most common virotype (34; 46.5%) was predominant among the subclade C1 population, while virotypes E and F (21; 28.7%) were detected among subclade C2, which had the highest VF scores and aminoglycoside resistance rates.ConclusionsThe appearance of virotypes E and F among subclade C2 strains with higher rates of aminoglycoside resistance/virulence gene content shows the shifting dynamics of this pandemic clone in response to antibiotic selection pressure by establishing subsets with higher survival potential.

Antimicrobial resistance surveillance, intensive care units of a tertiary care hospital, Islamabad

Abstract Background Hospital Acquired Infections (HAIs) are the emerging threat in the health care facilities around the globe. Most of Intensive Care Unit(ICU)patients are frequently on broad spectrum antimicrobials, this induces selective antibiotic pressure which leads to development of Antimicrobial Resistance (AMR) among the microorganisms of ICUs. Objective was to study the occurrence of different types of HAIs in patients admitted to various ICUs of and the AMR pattern of the bacterial pathogens isolated from them. Methods It was a retrospective descriptive study based on culture reports of the patients admitted to surgical, Medical ICU of tertiary care hospital during period from March 2019 to October 2019. Results Most common culture positive clinical specimen received was tracheal aspirate(35.3%) followed by exudate(22.7%).Pseudomonas from blood specimens were the most common organisms isolated; whereas Escherichia coli was the predominant organism found in urine, exudate and sterile fluid specimens. About 25% infections were HAIs, out of which pneumonia(7.24%)was the most common. Analysis of antimicrobial susceptibility pattern revealed that most of Gram-Negative Bacilli(GNB) was Multi Drug Resistant(MDR) resistant to three or more class of antibiotics such as cephalosporins, aminoglycosides, tetracyclines and fluoroquinolones. Conclusions Increasing trend of AMR among hospital acquired pathogens such as MDR-GNBs,MRSA and VRE pose a great threat to critically ill patients of ICUs. Study on AMR surveillance is the need of the hour as it helps the centers to generate local antibiogram which further helps in formulating the national data. Hence, such studies will be a stepping stone in establishing antimicrobial stewardship and regulate the antimicrobial resistance in future Key messages Study on AMR surveillance is the need of the hour. such studies will be a stepping stone in establishing antimicrobial stewardship and regulate the antimicrobial resistance in future.

Deciphering the mobility and bacterial hosts of antibiotic resistance genes under antibiotic selection pressure by metagenomic assembly and binning approaches

The presence of antibiotics can exert significant selection pressure on the emergence and spread of antibiotic resistance genes (ARGs) and antibiotic resistant bacteria (ARB). However, co-selection effects for ARGs, the mobility of ARGs and the identification of ARG hosts under high antibiotic selection pressures are poorly understood. Here, metagenomic assembly and binning approaches were used to comprehensively decipher the prevalence of ARGs and their potential mobility and hosts in activated sludge reactors treating antibiotic production wastewater. We found the abundance of different ARG types in antibiotic treatments varied greatly and certain antibiotic pressure promoted the co-selection for the non-corresponding types of ARGs. Antibiotic selection pressures significantly increased the abundance and proportions of ARGs mediated by plasmids (57.9%), which were more prevalent than those encoded in chromosomes (19.2%). The results indicated that plasmids and chromosomes had a tendency to carry different types of ARGs. Moreover, higher co-occurrence frequency of ARGs and MGEs revealed that antibiotics enhanced the mobility potential of ARGs mediated by both plasmids and integrative and conjugative elements. Among the 689 metagenome-assembled genomes (MAGs) with high estimated quality, 119 MAGs assigning to nine bacterial phyla were identified as the ARG hosts and 33 MAGs exhibited possible multi-resistance to antibiotics. Some ARG types tended to be carried by certain bacteria (e.g. bacitracin resistance genes carried by the family Burkholderiaceae) and thus showed a pronounced host-specific pattern. This study enhances the understanding of the mobility and hosts of ARGs and provides important insights into the risk assessment and management of antibiotic resistance.

Antibiotic-resistant Enterobacteriaceae from diseased freshwater goldfish.

Goldfish farming gained more attention among the ornamental fishes in aquaculture industry. The occurrence of bacterial infections and further antimicrobial treatment lead to the major crisis of antibiotic resistance in aquaculture. We have isolated diverse enterobacteriaceae groups which affect the goldfish and identified their response towards 46 antimicrobials of 15 different classes. Thirteen significant bacterial isolates such as Edwardsiella tarda, Serratia marcescens, Klebsiella aerogenes, Proteus penneri, P. hauseri, Enterobacter cloacae, E. cancerogenus, E. ludwigii, Citrobacter freundii, E. coli, Kluyvera cryocrescens, Plesiomonas shigelloides and Providencia vermicola were recovered from the infected fish with the Shannon-wiener diversity index of 2.556. Multiple antibiotic resistance (MAR) index was found to be maximum for P. penneri (0.87) and minimum for C. freundii and E. cloacae (0.22), highlighting the hyper antibiotic selection pressure in the farm. The minimum concentration of antibiotics required to inhibit most of the resistant isolates was found to be > 256 mcg/ml. All the isolates were susceptible towards ciprofloxacin. Plasmid curing and further AMR tests could reveal the location of antibiotic resistance genes mainly as plasmids which determine the large extent of AMR spread through horizontal gene transfer. This study is the first of its kind to investigate the antimicrobial resistance profile of enterobacteriaceae recovered from goldfish, before and after plasmid curing.

Effects of Antibiotics on the Bacterial Community, Metabolic Functions and Antibiotic Resistance Genes in Mariculture Sediments during Enrichment Culturing

The effect of antibiotics on the diversity and functioning of indigenous microorganisms in the environment has attracted much attention. In this study, effects of exposure to six different antibiotics on the bacterial community, metabolic functions and antibiotic resistance genes (ARGs) in marine sediments during enrichment culturing were investigated. Classical culture-dependent method and high-throughput 16S rRNA gene sequencing method were both applied. In the culture-dependent analysis, the obtained 1549 isolates belonged to four phyla (Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria) and 155 genera. Proteobacteria and Firmicutes were the dominant phyla. The diversity and abundance of obtained bacteria after antibiotic processing exhibited different degrees of decrease. Enrichment culturing for different time could also affect the bacterial community composition. Some genera of bacteria were not isolated in the control group, but they could be isolated in the antibiotic-treated groups. In high-throughput 16S rRNA gene amplicon sequencing analyses, all the effective reads were clustered into 2822 OTUs at 97% similarity cutoff; they were annotated to 49 phyla, 103 class, 220 orders, 347 families, 624 genera and 1122 species. An alpha diversity analysis indicated that the community diversity and richness decreased under antibiotic exposure. The changes at the genus level were much more obvious. Only 48 genera of 129 genera were shared by all the samples. A total of 29 genera which were not detected in the initial control sample could be detected in at least one antibiotic-treated group. SIMPER analysis showed that OTU2543 and OTU1450 were the most common taxa to the dissimilarity of bacterial community between antibiotic-treated groups and the control group. OTU2034 and OUT2543 were the most contributive taxa to dissimilarity of groups incubating for different time. Metabolism was the predominant bacterial function. A total of 30 ARGs were detected in the samples. This study mainly focused on the changes of microbiota under the selective pressure of antibiotics for different time and the results demonstrated that the antibiotic could affect the bacterial diversity and richness in the marine ecosystem.

Emerging nanobiomaterials against bacterial infections in postantibiotic era

AbstractAlthough numerous antibiotics have been developed and applied in clinic, bacterial infections are still serious threats to human health, due to the rapidly growing antibiotic resistance all over the world. The causes of antibiotic resistance include two main aspects: the formation of bacterial biofilms and the self‐evolution of bacteria under the antibiotic selection pressure. It is of great significance to develop effective strategies to treat the bacterial infections with serious antibiotic resistance. With excellent performance, such as size effect, specific physicochemical property, easy modification, and so on, nanomaterials exhibit enormous potential as enhancers or therapeutic agents to treat severe drug‐resistant bacterial infections. In this review, the underlying causes for the antibiotic resistance are fully summarized and discussed. Subsequently, the promising therapeutic methods by using nanomaterials are provided and discussed to treat bacterial infections with serious antibiotic resistance.

Correlation between Exogenous Compounds and the Horizontal Transfer of Plasmid-Borne Antibiotic Resistance Genes.

The global spread of antibiotic resistance has posed a serious threat to public healthcare and undermined decades of progress made in the fight against bacterial infections. It has been demonstrated that the lack of novel effective antibiotics and rapid spread of antibiotic resistance genes via horizontal transfer in the ecosystem are mainly responsible for this crisis. Notably, plasmid-mediated horizontal transfer of antibiotic resistance genes (ARGs) is recognized as the most dominant dissemination pathway of ARGs in humans, animals and environmental settings. Antibiotic selective pressure has always been regarded as one of the crucial contributors to promoting the dissemination of antibiotic resistance through horizontal gene transfer (HGT). However, the roles of exogenous compounds and particularly non-antibiotic drugs in the spread of ARGs are still underappreciated. In this review, we first summarize the major pathways of HGT in bacteria, including conjugation, transformation, transduction and vesiduction. Subsequently, an overview of these compounds capable of promoting the HGT is presented, which guides to the formulation of more reasonable dosing regimens and drug residue standards in clinical practice. By contrast, these compounds that display an inhibition effect on HGT are also highlighted, which provides a unique strategy to minimize the spread of ARGs. Lastly, we discuss the implementations and challenges in bringing these HGT inhibitors into clinical trials.

Variation and distribution of antibiotic resistance genes and their potential hosts in microbial electrolysis cells treating sewage sludge

Microbial electrolysis cells (MECs) system is an emerging pollution control technology. However, information on the variation of antibiotic resistance genes (ARGs) in MECs treating sewage sludge is still very limited. In this study, the fate of ARGs and their correlation with microbes in MECs under different applied voltages (0–1.5 V) were studied. Most target ARGs were effectively removed, but tetB, tetM and tetQ were enriched up to 2.05 log units in suspended sludge. Most ARGs were mainly distributed on electrodes, except tetQ and tetM enriched in suspended sludge. The selective pressure of residual antibiotics in the sewage sludge was negligible. Horizontal gene transfer was validated for the spread of sul1, sul2, tetA and tetC in MECs. Network analysis revealed that the potential hosts of ARGs mainly belonged to Bacteroidetes, Firmicutes and Proteobacteria. Some genera related to electron transfer were newly found to be the potential ARGs hosts in MECs.