Antibiotics

19 pages, 2941 KB

Open AccessArticle

Inhaled Ciprofloxacin as an Alternative Treatment for Infection with Coxiella burnetii

by Rachel E. Ireland, Kevin R. Bewley, M. Gill Hartley, Karleigh A. Hamblin, Stuart J. Armstrong, Michelle Nelson, Thomas R. Laws, Isobel H. Norville, Francisco J. Salguero, James D. Blanchard, Francis Dayton, Igor Gonda, Helen S. Atkins and Sarah V. Harding

Antibiotics 2026, 15(3), 293; https://doi.org/10.3390/antibiotics15030293 - 13 Mar 2026

Abstract

Background/Objectives: Q fever, caused by Coxiella burnetii, is typically treated with doxycycline, but its efficacy is limited in chronic cases and may be poorly tolerated. Systemic ciprofloxacin shows limited activity for acute Q fever. However, inhaled liposomal formulations may provide therapeutic benefit. [...] Read more.

Background/Objectives: Q fever, caused by Coxiella burnetii, is typically treated with doxycycline, but its efficacy is limited in chronic cases and may be poorly tolerated. Systemic ciprofloxacin shows limited activity for acute Q fever. However, inhaled liposomal formulations may provide therapeutic benefit. Methods: Two inhaled ciprofloxacin formulations (Lipoquin^® and Apulmiq^®) were evaluated in an A/J mouse model of Q fever and compared with intraperitoneal ciprofloxacin and oral doxycycline. Initially, pharmacokinetic studies were performed to determine an appropriate dosing regimen for the inhaled ciprofloxacin formulations. A separate cohort of mice were then infected with C. burnetii and treated once daily via nebulisation with Lipoquin or Apulmiq, initiated at 24, 48, or 72 h post-challenge. Clinical signs, weight change, splenomegaly, bacterial burden, and lung histopathology were evaluated. Results: Pharmacokinetic analysis confirmed sustained lung concentrations of inhaled ciprofloxacin, supporting once-daily dosing. Inhaled Lipoquin and Apulmiq significantly reduced clinical signs, weight loss, splenomegaly, and pulmonary bacterial burden compared to untreated controls and doxycycline-treated mice. Histopathology revealed decreased lung inflammation and lesion severity following inhalational dosing. Systemic ciprofloxacin slightly reduced splenic bacterial burden but was less effective in controlling pulmonary infection. Conclusions: Inhaled liposomal ciprofloxacin demonstrated superior protection and reduced respiratory manifestations of Q fever compared to doxycycline and systemic ciprofloxacin. These findings suggest inhaled formulations may represent a viable alternative for the treatment of Q fever pneumonia. Further studies are needed to evaluate clinical applicability and long-term outcomes. Full article

(This article belongs to the Section Novel Antimicrobial Agents)

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15 pages, 1269 KB

Open AccessArticle

Emergence of Clinically Macrolide-Unresponsive Mycoplasma pneumoniae Segmental/Lobar Pneumonia and COVID-19 Pneumonia in Children in Taiwan, 2024–2025

by Hao-Yuan Lee, Chien-Chin Chen, Shu-Hua Ko, Yu-Ling Huang, En-Pen Chang, Cheng-Yi Hsu, Jia Ru Wu, Wei-Hsin Chen, Yu-Chau Hsu, Meng-Yen Li, Yu-Lung Hsu, Wen-Yuan Lee and Chyi-Liang Chen

Antibiotics 2026, 15(3), 292; https://doi.org/10.3390/antibiotics15030292 - 13 Mar 2026

Abstract

Background: To date, no study has compared the clinical characteristics of Mycoplasma pneumoniae-associated segmental/lobar pneumonia, Mycoplasma bronchopneumonia, and COVID-19 pneumonia primarily caused by the NB.1.8.1 variant in children. Methods: We examined the epidemiologic trends of pneumonia, segmental/lobar pneumonia, and COVID-19 pneumonia [...] Read more.

Background: To date, no study has compared the clinical characteristics of Mycoplasma pneumoniae-associated segmental/lobar pneumonia, Mycoplasma bronchopneumonia, and COVID-19 pneumonia primarily caused by the NB.1.8.1 variant in children. Methods: We examined the epidemiologic trends of pneumonia, segmental/lobar pneumonia, and COVID-19 pneumonia at a teaching hospital from 2015 to 2025. In addition, we compared the clinical characteristics of children hospitalized with Mycoplasma segmental/lobar pneumonia, Mycoplasma bronchopneumonia, and COVID-19 pneumonia during the NB.1.8.1 variant wave in 2024–2025. Results: Between 2015 and 2024, 10,601 pneumonia cases were identified, including 525 cases of segmental/lobar pneumonia and 162 cases of COVID-19 pneumonia. An outbreak of segmental/lobar M. pneumoniae pneumonia and COVID-19 pneumonia occurred in Taiwan during 2024–2025. Starting in early 2025, monthly Mycoplasma positivity rates among children with segmental/lobar pneumonia and bronchopneumonia exceeded 60%. Mycoplasma pneumonia predominantly affected children aged 6–11 years, whereas COVID-19 pneumonia mainly occurred in those younger than 3 years of age. Fever, cough, and rhinorrhea were the most common symptoms in all groups, limiting clinical differentiation. Children with segmental/lobar Mycoplasma pneumonia were more likely to present with prolonged fever (>5 days), lymphocytopenia, a neutrophil-to-lymphocyte ratio (NLR) ≥ 3, and elevated C-reactive protein (CRP) levels, each of which was strongly associated with macrolide non-response (all p < 0.001). Conclusions: Children with segmental/lobar Mycoplasma pneumonia demonstrated more severe clinical manifestations. Segmental/lobar involvement and inflammatory markers, such as lymphocytopenia, elevated NLR, and increased CRP levels, were associated with macrolide non-response. These indicators may help guide therapeutic decision-making in pediatric M. pneumoniae pneumonia. Full article

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17 pages, 2383 KB

Open AccessArticle

Deficiency of the Mycobacterial Lipoarabinomannan Biosynthesis Glycosyltransferase MptC Enhances Antibacterial Immune Response and Rifapicin Antibiotic Susceptibility

by Jiaxin Hu, Hongliang Chen, Zhongkun Li, Hao Sun, Yi-Cheng Sun and Xiao-Lian Zhang

Antibiotics 2026, 15(3), 291; https://doi.org/10.3390/antibiotics15030291 - 13 Mar 2026

Abstract

Background/Objectives: The mycobacterial complex cell envelope serves as a formidable barrier against host immunity and antibiotics. Lipomannan (LM) and lipoarabinomannan (LAM) are key structural components of the mycobacterial envelope and potent immunomodulators. The mycobacterial lipoarabinomannan biosynthesis mannosyltransferase MptC modifies the multiple α-(1→2)-linked branched [...] Read more.

Background/Objectives: The mycobacterial complex cell envelope serves as a formidable barrier against host immunity and antibiotics. Lipomannan (LM) and lipoarabinomannan (LAM) are key structural components of the mycobacterial envelope and potent immunomodulators. The mycobacterial lipoarabinomannan biosynthesis mannosyltransferase MptC modifies the multiple α-(1→2)-linked branched mannan residues of LAM in the mycobacteria. However, the role of MptC in mycobacterial infectivity, antibiotic susceptibility and host immune regulation remains poorly understood. Methods: An mptC (also named MSMEG_4247) knockout Mycobacterium smegmatis mc²-155 (M. smeg) strain (designated as M. smegΔmptC) was generated using CRISPR–Cas12a technology. The effects of MptC on bacterial physiology, cell wall permeability, drug sensitivity, immune cell function, and survival during infection are analyzed through glycogen staining, drug sensitivity tests, and cellular and mouse infection models. Results: MptC deficiency results in a loss of LM and increase in LAM synthesis. The M. smegΔmptC mutant strain exhibits enhanced cell wall permeability and reduces hydrophobicity. Functionally, the mptC knockout strain increases the intracellular cytokines (IFN-γ, TNF-a and IL-17) production of T cells in mice. Consequently, results based on both macrophage and mouse infection models demonstrate that the M. smegΔmptC strain has less bacterial loads and higher susceptibility to antibiotic rifampicin. Conclusions: Mannosyltransferase MptC plays an important role in maintaining cell wall integrity (via LM/LAM synthesis), regulating T cell responses, and influencing antibiotic susceptibility in mycobacteria. Full article

(This article belongs to the Special Issue Rethinking Tuberculosis Treatment: Integrating Antimicrobial Compounds, Host Responses, and Comorbidities)

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10 pages, 571 KB

Open AccessArticle

Screening and Cohorting of CRE Patients: The Strategic Role of Bed Management in a Monocentric Pre–Post Observational Study

by Salvatore Altavilla, Nicoletta Di Pietro, Daniela Loconsole and Francesco Di Gennaro

Antibiotics 2026, 15(3), 290; https://doi.org/10.3390/antibiotics15030290 - 12 Mar 2026

Abstract

Background: Carbapenem-resistant Enterobacterales (CRE) require early identification and appropriate patient placement to prevent in-hospital transmission. Bed Management plays a key organizational role in coordinating screening results and isolation strategies; however, evidence on its impact on patient flow and isolation practices remains limited. Aim: [...] Read more.

Background: Carbapenem-resistant Enterobacterales (CRE) require early identification and appropriate patient placement to prevent in-hospital transmission. Bed Management plays a key organizational role in coordinating screening results and isolation strategies; however, evidence on its impact on patient flow and isolation practices remains limited. Aim: To evaluate whether the implementation of a Bed Management-coordinated structured CRE screening pathway was associated with changes in patient placement appropriateness and time to admission (TTA). Methods: We conducted a retrospective cohort study including all patients with a positive rectal swab for CRE during two study periods (PRE: 2024; POST: 2025) in a tertiary care hospital. A structured CRE screening pathway coordinated by Bed Management was implemented in the POST period. Primary outcomes were cohort isolation rates and TTA. Continuous variables were compared using the Mann–Whitney U test and categorical variables using the chi-square test. Results: A total of 158 CRE-positive patients were included (69 in the PRE period and 89 in the POST period). Patient characteristics were comparable between periods (median age 75 years [IQR 64–81] vs. 73 years [IQR 64–82]; female sex 33.3% vs. 44.9%, p = 0.189). Cohort isolation rates were higher in the POST period. Median time to admission (TTA) decreased from 74.8 h (IQR 47.2–124.6) in PRE to 70.7 h (IQR 35.1–139.9) in POST; however, this difference did not reach statistical significance (Mann–Whitney U test, p = 0.630). Conclusions: A Bed Management-coordinated CRE screening pathway was associated with improved cohort isolation practices and an observed, non-significant reduction in TTA. These findings suggest that integrating infection prevention workflows with centralized bed allocation may be feasible without adversely affecting admission timeliness. Further studies with larger samples and longer observation periods are warranted. Full article

(This article belongs to the Special Issue Antibiotics in Pathogen Control: From Infection to Antimicrobial Resistance)

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13 pages, 241 KB

Open AccessArticle

Critical Analysis of Fixed-Dose Antibiotic Combinations Sold in Kinshasa—Democratic Republic of the Congo

by Jocelyn Kakumba Mankulu, Dadit Kitenge Ive, Freddy Mugisho Kasago, Exaucé Mpuya Mpuya, Bertin K. Mfuamba, Jean-Pierre Mufusama Koy Sita, Patient Ciza Hamuli, Trésor Kimbeni Malongo, Jérémie Mbinze Kindenge, Jean-Marie Liesse Iyamba and Didi Mana Kialengila

Antibiotics 2026, 15(3), 289; https://doi.org/10.3390/antibiotics15030289 - 12 Mar 2026

Abstract

Background: Fixed-dose combination drugs (FDCs) are combinations of two or more active ingredients in a single dosage form. These formulations have proven effective in combating the development of resistance in diseases such as tuberculosis and malaria. Despite the benefits observed in the [...] Read more.

Background: Fixed-dose combination drugs (FDCs) are combinations of two or more active ingredients in a single dosage form. These formulations have proven effective in combating the development of resistance in diseases such as tuberculosis and malaria. Despite the benefits observed in the aforementioned cases, fixed-dose antibiotics combinations (FDACs) are increasingly raising questions about their rationality. This is the case for several FDACs listed in the AWaRe classification as not recommended, which unfortunately remain available on the pharmaceutical market, particularly in low- and middle-income countries like the Democratic Republic of Congo (DRC). Objectives: To identify the essential medicines available in pharmacies open to the public in the city of Kinshasa and to assess their inclusion in the DRC’s National List of Essential Medicines (NLEM) and in the World Health Organization’s (WHO) List of Essential Medicines (LEM). The rationality of the FDACs circulating in the city of Kinshasa were also evaluated based on the 2023 AWaRe classification. Methods: A cross-sectional and descriptive study was conducted between February and October 2025 in Kinshasa. For this purpose, fifty registered pharmacies open to the public were selected by systematic random sampling as the research sample. Data collection consisted of completing a data collection form after we had provided the pharmacies’ owners with the necessary explanations regarding the importance of the study and guaranteed their anonymity. Results: The controlled FDACs encountered comprised 27 specialties across 15 different formulations. Out of 15 formulations, 12 (80%) were included on the WHO list of non-recommended antibiotics and were not included in the DRC’s NLEM nor in the WHO’s LEM. Some had been withdrawn from the market in their countries of manufacture. Of the 15 FDACs evaluated for their rationality and compliance, the injectable FDACs presented problems related to the relevance and completeness of information contained on their packaging. On their primary packaging, there was a significant difference in the expiration dates of the powder and sterile water for injection contained in the combination pack, ranging from 6 to 36 months. Furthermore, the secondary packaging lacked data related to the sterile water for injection contained in the combination pack. In addition, several medications contained the same therapeutic combination. For injectable FDAC, for example, the combination Ceftriaxone-Sulbactam was represented by eight medications. For oral FDACs, the combination Sulfamethoxazole-Trimethoprim was represented by seven medications. Globally, 100% of these drug combinations originated from India. Conclusions: Fifteen varieties of FDACs were available in Kinshasa, most of which (80%) were unsuitable. It is important that public health authorities address this situation and develop stricter guidelines for granting marketing authorizations, particularly for FDACs. Full article

(This article belongs to the Special Issue Antimicrobial Stewardship—from Projects to Standard of Care)

18 pages, 696 KB

Open AccessArticle

Analysis of Antibiotic Consumption Trends and Pathogens’ Epidemiological Profile Within a Multidisciplinary Clinical Hospital from Romania

by Andreea-Roxana Ungureanu, Andreea-Alina Dumitru, Emma-Adriana Ozon, Andrei-Tudor Rogoz, Raluca-Narcisa Anghel, Elena Ciucu, Ancuța-Cătălina Fița and Nicoleta-Mirela Blebea

Antibiotics 2026, 15(3), 288; https://doi.org/10.3390/antibiotics15030288 - 12 Mar 2026

Abstract

Background/Objectives: In the broad and current context of antimicrobial resistance, antibiotic management and therapeutic surveillance are essential in hospitals. The present study (five-year retrospective, 2020–2024) aimed to analyze antibiotic consumption in relation to pathogens identified in a multidisciplinary hospital. Results: In terms of [...] Read more.

Background/Objectives: In the broad and current context of antimicrobial resistance, antibiotic management and therapeutic surveillance are essential in hospitals. The present study (five-year retrospective, 2020–2024) aimed to analyze antibiotic consumption in relation to pathogens identified in a multidisciplinary hospital. Results: In terms of antibiotic consumption (overall 2020–2024), although initially Watch antibiotics were predominantly used, a decrease was observed in favor of Access class antibiotics (sharply increase from 2022 to 2023 and maximum in 2024). For Reserve antibiotics, only slight annual fluctuations were observed, but there was an important reduction in colistin consumption. The most used were cephalosporins (cefazolin, cefuroxime and ceftriaxone), carbapenems (meropenem and ertapenem), vancomycin and linezolid. Regarding pathogens, the most notable were: Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus spp., Pseudomonas aeruginosa. Among the ESKAPE bacteria, Acinetobacter baumannii was the least frequent in our samples. ESKAPE bacteria predominantly colonized specimens from the respiratory tract, digestive tract, skin and soft tissue. Resistant strains were observed, mainly Methicillin-resistant Staphylococcus aureus (MRSA) and Extended-Spectrum Beta-Lactamase (ESBL) Klebsiella spp., but no alarming increases in number were recorded in the analyzed period. Methods: The analysis was carried out using tools recommended by the World Health Organisation (Access Watch Reserve antibiotics classification (AWaRe); Bacterial Priority Pathogen List (BBPL); Defined Daily Dose (DDD)), Average Annual Percent Change (AAPC) calculation and ESKAPE classification (bacteria group: Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter spp.). Conclusions: Relatively stable trends in bacterial isolates and resistant strains over five years (2020–2024) are consistent with effective antimicrobial stewardship practices. Full article

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19 pages, 2727 KB

Open AccessArticle

Plasmid-Driven Resistome Diversity in 9700 Escherichia coli Genomes Across Phylogroups and Sequence Types

by Adel Azour, Ghassan M. Matar and Melhem Bilen

Antibiotics 2026, 15(3), 287; https://doi.org/10.3390/antibiotics15030287 - 12 Mar 2026

Abstract

Background/Objectives: Plasmids are key vehicles for the dissemination of antimicrobial resistance (AMR), yet their contribution to the global resistome architecture of Escherichia coli remains poorly resolved. This study aimed to quantify how plasmid backbones shape the distribution, mobility, and stabilization of resistance [...] Read more.

Background/Objectives: Plasmids are key vehicles for the dissemination of antimicrobial resistance (AMR), yet their contribution to the global resistome architecture of Escherichia coli remains poorly resolved. This study aimed to quantify how plasmid backbones shape the distribution, mobility, and stabilization of resistance genes across diverse phylogenetic backgrounds. Methods: We analyze 9700 high-quality genomes spanning major phylogroups and sequence types. Plasmidome reconstruction was integrated with lineage-resolved antimicrobial resistance gene (ARG) mapping to characterize plasmid–ARG associations and evolutionary patterns. Results: Although most antimicrobial resistance genes (ARGs) are chromosomal, plasmids disproportionately encode clinically important determinants including bla_NDM-5, mcr-1.1, and multiple bla_CTX-M alleles that show strong, recurrent associations with a restricted set of backbone families, most notably IncX3, IncX4, IncI, and IncF. These conserved plasmid–gene modules recur across phylogenetic backgrounds and continental scales. We identify a marked divergence in evolutionary strategies: generalist phylogroups (A, B1, D) maintain plasmid-rich and highly diverse resistomes, whereas globally dominant Extraintestinal Pathogenic E. coli (ExPEC) clones such as ST131 and ST410 exhibit reduced plasmid dependency and frequent chromosomal integration of extended-spectrum β-lactamase (ESBL) genes, particularly bla_CTX-M-15, consistent with a shift toward vertically stabilized resistomes. By integrating plasmidome reconstruction with lineage-resolved ARG mapping, this study delivers the most extensive plasmid-focused resistome analysis to date, revealing highly modular plasmid–ARG networks structured around a small number of high-risk backbone types. These backbones account for the majority of globally relevant ARGs, including 64.6% of bla_NDM-5 and 76.4% of mcr-1.1 detections. Conclusions: Together, our findings establish plasmid lineages rather than individual genes or clones as central units of AMR dissemination and critical targets for future genomic surveillance and intervention strategies. Full article

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17 pages, 354 KB

Open AccessArticle

Multicenter Analytical Performance Evaluation of the BD Phoenix NMIC-461 Panel for Carbapenemase Classification and Antimicrobial Susceptibility Testing of Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter spp.

by Jingjia Zhang, Liying Sun, Ge Zhang, Wei Kang, Tong Wang, Jin Li, Haotian Gao, Qiwen Yang, Kuixia Sun, Qian Wang and Hongli Sun

Antibiotics 2026, 15(3), 286; https://doi.org/10.3390/antibiotics15030286 - 12 Mar 2026

Abstract

Objectives: To evaluate the capability of the BD Phoenix NMIC-461 panel in the detection and classification of carbapenemase production and antimicrobial susceptibility testing of 10 antimicrobial agents among Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter spp. Methods: A total of 714 non-repetitive clinical [...] Read more.

Objectives: To evaluate the capability of the BD Phoenix NMIC-461 panel in the detection and classification of carbapenemase production and antimicrobial susceptibility testing of 10 antimicrobial agents among Enterobacterales, Pseudomonas aeruginosa, and Acinetobacter spp. Methods: A total of 714 non-repetitive clinical isolates from three tertiary hospitals in China were enrolled. Carbapenemase production was confirmed by the modified carbapenem inactivation method (mCIM), while carbapenemase typing was validated by polymerase chain reaction (PCR) and Sanger sequencing. Antimicrobial susceptibility testing (AST) for ten antimicrobial agents was performed using broth microdilution (BMD) as the reference method. Results: The sensitivity and specificity of carbapenemase detection were 98.8% (95% CI, 96.6–99.6) and 92.4% (95% CI, 89.5–94.6) separately compared to sequencing. Classification accuracy was compromised by carbapenemase-positive unclassified strains, particularly reducing sensitivity for Enterobacterales. Excluding unclassified strains, the sensitivity and specificity were: for class A, 100% (95% CI, 94.0–100) and 97.3% (95% CI, 95.6–98.4); for class B, 97.1% (95% CI, 89.7–99.2) and 97.6% (95% CI, 96.0–98.6); and for class D, 94.0% (95% CI, 87.9–97.3) and 99.1% (95% CI, 97.8–99.7). The panel was subject to limitations for carbapenemase detection when applied to Pseudomonas aeruginosa. The NMIC-461 panel demonstrated excellent performance for ten BMD-evaluated agents across four bacterial categories, with essential agreement (EA) exceeding 95% and category agreement (CA) exceeding 90% except for Levofloxacin, and major error (ME) and very major error (VME) rates below 3% and 1.5%, respectively. Conclusions: The BD Phoenix NMIC-461 panel provides reliable AST results for commonly encountered Gram-negative bacterial isolates. Regarding carbapenemase detection, the panel demonstrates high sensitivity but only moderate specificity in classifying carbapenemase-producing organisms (CPO), with a relatively high proportion of positive unclassified isolates among Enterobacterales and low specificity for P. aeruginosa. Overall, the implementation of NMIC-461 testing holds promise for significantly reducing turnaround time in both carbapenemase detection and classification. Full article

(This article belongs to the Special Issue Multi-Drug Resistant Gram-Negative Infections: Molecular Epidemiology, Microbiological Diagnosis, and Antimicrobial Treatment, 2nd Edition)

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15 pages, 1136 KB

Open AccessArticle

MYCOPLASMA IST3 Results and Antimicrobial Susceptibility in PCR-Positive Urine Samples for Ureaplasma spp.

by Rukiye Berkem and Tuğçe Özyol Atkaya

Antibiotics 2026, 15(3), 285; https://doi.org/10.3390/antibiotics15030285 - 11 Mar 2026

Abstract

Background: Ureaplasma spp. and Mycoplasma hominis are urogenital pathogens that may be missed by routine culture, particularly in patients with genitourinary symptoms in whom conventional methods fail to identify an etiologic agent. Limited routine implementation of targeted diagnostics and antimicrobial susceptibility testing (AST) [...] Read more.

Background: Ureaplasma spp. and Mycoplasma hominis are urogenital pathogens that may be missed by routine culture, particularly in patients with genitourinary symptoms in whom conventional methods fail to identify an etiologic agent. Limited routine implementation of targeted diagnostics and antimicrobial susceptibility testing (AST) for these organisms may contribute to diagnostic uncertainty and treatment failure. Methods: Seventy-five midstream urine samples submitted for suspected urinary tract infection and positive for Ureaplasma spp. according to a q-PCR urinary panel (Bioeksen, İstanbul, Türkiye) were tested the same day with MYCOPLASMA IST3 (bioMérieux, Marcy-l’Étoile, France) to assess growth and antimicrobial susceptibility. Results: q-PCR detected U. parvum in 54/75 (72%), U. urealyticum in 15/75 (20%), and both species in 6/75 (8%); M. hominis was not included in the PCR panel. MYCOPLASMA IST3 showed growth in 70/75 samples (positive percent agreement, 93.33%), while 5/75 (discordance, 6.66%) showed no growth. Among culture-positive samples, 57/70 (81.42%) yielded Ureaplasma spp. alone, and 13/70 (18.58%) yielded Ureaplasma spp. together with M. hominis. Resistance to levofloxacin and tetracycline was observed in 15.7% and 12.9% of Ureaplasma spp. isolates, respectively; resistance to moxifloxacin, erythromycin, and telithromycin was observed in 2.9% of isolates for each agent. In M. hominis isolates, no resistance to levofloxacin, moxifloxacin, or tetracycline was observed, whereas clindamycin resistance was observed in 7.7% of isolates. Conclusions: In addition to intrinsic resistance, acquired antimicrobial resistance in Ureaplasma and Mycoplasma species appears to be increasing; therefore, treatment decisions should be guided by AST whenever feasible. Clinical laboratories should implement appropriate diagnostic methods for these organisms and perform susceptibility testing when indicated to support clinical decision making and optimize antimicrobial selection. Full article

(This article belongs to the Special Issue Advancing and Standardising Antimicrobial Susceptibility and Resistance Detection Methodology)

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5 pages, 488 KB

Open AccessCorrection

Correction: McSorley, J.C. Analysis of ESAC-Net/EARS-Net Data from 29 EEA Countries for Spatiotemporal Associations Between Antimicrobial Use and Resistance—Implications for Antimicrobial Stewardship? Antibiotics 2025, 14, 399

by James C. McSorley

Antibiotics 2026, 15(3), 284; https://doi.org/10.3390/antibiotics15030284 - 11 Mar 2026

Abstract

1 [...] Full article

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24 pages, 22713 KB

Open AccessArticle

Revitalizing Trimethoprim/Sulfamethoxazole via Nanotechnology for Improved Pharmacokinetics and Antibacterial Efficacy

by Yaxin Zhou, Jing Xu, Guonian Dai, Bing Li, Weiwei Wang, Bintao Zhai, Shulin Chen and Jiyu Zhang

Antibiotics 2026, 15(3), 283; https://doi.org/10.3390/antibiotics15030283 - 10 Mar 2026

Abstract

Objective: The therapeutic efficacy of the classic antibiotic combination trimethoprim/sulfamethoxazole (TMP/SMZ) is often limited by the significant pharmacokinetic mismatch. In this study, a polyethylene glycol-polylactic-co-glycolic acid (PEG-PLGA) nanodelivery system was employed to improve the pharmacokinetic matching of TMP and SMZ. The investigation [...] Read more.

Objective: The therapeutic efficacy of the classic antibiotic combination trimethoprim/sulfamethoxazole (TMP/SMZ) is often limited by the significant pharmacokinetic mismatch. In this study, a polyethylene glycol-polylactic-co-glycolic acid (PEG-PLGA) nanodelivery system was employed to improve the pharmacokinetic matching of TMP and SMZ. The investigation also evaluated the enhanced in vivo antibacterial efficacy of this formulation. Methods: Ultra-High Performance Liquid Chromatography–Tandem Mass Spectrometry (UPLC-MS/MS) was employed to systematically characterize the absorption, distribution, and excretion profiles of PEG-PLGA-loaded TMP nanoparticles (NPs) in rats. In vitro antibacterial activity was assessed against Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus). In vivo efficacy and biosafety of the TMP NPs/SMZ regimen were evaluated using a murine E. coli infection model via survival monitoring, biochemical assays, and histopathology. Results: Pharmacokinetic analysis revealed that TMP NPs achieved a relative bioavailability of 193.05% and extended the elimination half-life by 3.37-fold compared to free TMP. Tissue distribution showed significantly increased drug accumulation in the liver, spleen, and kidneys, with renal clearance as the primary excretion pathway (73.89%). In vitro, the nano-formulation reduced the minimum inhibitory concentration (MIC) by 2-4-fold and shortened the bactericidal duration from 12 to 8 h. In vivo, the TMP NPs/SMZ combination significantly improved survival rates, accelerated recovery, and alleviated infection-induced organ damage without systemic toxicity. Conclusions: This nanotechnology-based strategy effectively aligns the pharmacokinetics of TMP and SMZ, prolongs their synergistic window, and enhances biosafety, offering a viable approach to revitalize classic antibiotic combinations. Full article

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18 pages, 570 KB

Open AccessArticle

Occurrence of Antimicrobial Resistance in Indicator Bacteria and Campylobacter spp. Isolated from Commercial Raw-Meat-Based Food for Dogs and Cats in Belgium

by Junjia He, Ilias Chantziaras, Cristina Garcia-Graells, Moniek Ringenier, Suzanne Dewulf, Filip Boyen, Jeroen Dewulf and Cécile Boland

Antibiotics 2026, 15(3), 282; https://doi.org/10.3390/antibiotics15030282 - 10 Mar 2026

Abstract

Background: Raw-meat-based diets (RMBDs) for companion animals have gained popularity but may serve as vehicles for antimicrobial-resistant (AMR) bacteria, posing risks to animal and public health. This study investigated the occurrence and risk factors of AMR in indicator bacteria (Escherichia coli, [...] Read more.

Background: Raw-meat-based diets (RMBDs) for companion animals have gained popularity but may serve as vehicles for antimicrobial-resistant (AMR) bacteria, posing risks to animal and public health. This study investigated the occurrence and risk factors of AMR in indicator bacteria (Escherichia coli, Enterococcus faecalis, Enterococcus faecium) and Campylobacter spp. from commercial RMBD products. Methods: In 2023, 50 RMBD samples were collected in Belgium, representing 21 brands from five countries. After both selective and non-selective isolation and MALDI-TOF identification, antimicrobial susceptibility testing of the isolates was performed using broth microdilution. Results: From non-selective media, E. coli was found in 45 samples (90.0%), E. faecalis in 31 samples (62.0%), E. faecium in 23 samples (46.0%), and Campylobacter spp. in 3 samples (6.0%). Among these, one E. faecalis strain with acquired resistance to vancomycin and daptomycin was isolated. Multidrug resistance (MDR) was identified in 17 isolates from 15 samples (30.0%), including 14 MDR E. coli, 1 MDR E. faecalis, and 2 MDR E. faecium. From selective media, presumptive ESBL/AmpC-producing E. coli were detected in 17 samples (34.0%), and 5 E. faecium from linezolid-supplemented media were confirmed by the broth microdilution method. Samples from Belgian origin showed significantly higher E. faecium prevalence (76.5%) compared to Dutch samples (21.4%) (OR = 11.9, p < 0.001). Minor livestock sources were associated with increased MDR risk (OR = 5.52, p = 0.016). Conclusions: Commercial RMBDs in Belgium exhibit widespread bacterial contamination with concerning AMR patterns. These findings highlight the need for improved production standards in the RMBD industry and the need to raise awareness in pet owners. Full article

(This article belongs to the Special Issue Risks and Current Knowledge of Antimicrobial Resistance in Companion Animals)

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23 pages, 1106 KB

Open AccessReview

Revisiting the Fight Against Acinetobacter baumannii: Emerging Non-Antibiotic Strategies

by Victor Hugo Montini, Laura Santana Buso, Pedro Henrique Takata, Gabriel Henrique Maximino Santos, Bruna Carolina Gonçalves, Thiago Hideo Endo, Mariana Homem de Mello Santos, Eliana Carolina Vespero, Renata Katsuko Takayama Kobayashi and Gerson Nakazato

Antibiotics 2026, 15(3), 281; https://doi.org/10.3390/antibiotics15030281 - 10 Mar 2026

Abstract

This review discusses emerging in vitro and in vivo strategies for the control of Acinetobacter baumannii, a critical multidrug-resistant pathogen; the increasing isolation of strains resistant to multiple drugs, including newly developed and last-resort antibiotics, has highlighted the urgent need to pursue [...] Read more.

This review discusses emerging in vitro and in vivo strategies for the control of Acinetobacter baumannii, a critical multidrug-resistant pathogen; the increasing isolation of strains resistant to multiple drugs, including newly developed and last-resort antibiotics, has highlighted the urgent need to pursue adjunctive therapeutic technologies. The article aims to provide an overview of alternative control approaches beyond conventional antibiotics. Emphasis is placed on strategies based on the disruption of essential metabolic pathways, nanotechnology-based approaches such as antibiotic-coated nanoparticles, in vivo bacteriophage therapy, and drug repurposing, specifically compounds such as selective serotonin reuptake inhibitors (SSRIs), as a means of exploiting already approved pharmaceuticals. By synthesizing recent findings, this review highlights current advances in the development of innovative therapeutic strategies against A. baumannii infections. Full article

(This article belongs to the Topic Antimicrobial Agents and Nanomaterials—2nd Edition)

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15 pages, 676 KB

Open AccessArticle

Pentoxifylline and Tocopherol for the Management of Medication-Related Osteonecrosis of the Jaw (MRONJ): A Retrospective Clinical Audit

by Niccolò Lombardi, Virina Basta, Chiara Morelli, Giulia Ghidini, Giovanni Lodi and Elena M. Varoni

Antibiotics 2026, 15(3), 280; https://doi.org/10.3390/antibiotics15030280 - 10 Mar 2026

Abstract

Background/Objectives: Medication-related osteonecrosis of the jaw (MRONJ) is a challenging complication in patients receiving antiresorptive therapy. Management strategies range from conservative pharmacological approaches to extensive surgical resection of necrotic bone. This clinical audit retrospectively evaluated the clinical outcomes of patients undergoing sequestrectomy [...] Read more.

Background/Objectives: Medication-related osteonecrosis of the jaw (MRONJ) is a challenging complication in patients receiving antiresorptive therapy. Management strategies range from conservative pharmacological approaches to extensive surgical resection of necrotic bone. This clinical audit retrospectively evaluated the clinical outcomes of patients undergoing sequestrectomy for MRONJ, comparing those treated with antibiotics alone with those receiving antibiotics in combination with the pentoxifylline–tocopherol (PENTO) protocol. The PENTO protocol was introduced at our institution in 2021 and has since been routinely prescribed for all MRONJ patients. Methods: We analyzed 92 MRONJ sites treated with sequestrectomy. Conservative management consisted of antibiotic therapy, with or without adjunctive PENTO (pentoxifylline 800 mg/day and tocopherol 800 IU/day, administered both preoperatively and postoperatively). The primary outcome was healing at three months post-surgery, while the secondary outcome was disease recurrence during longer-term follow-up. Results: Complete healing was achieved in 56 of the 92 sites, with a mean follow-up of 9.98 ± 12.76 months among healed cases. No statistically significant differences in healing rates were observed between the PENTO and antibiotic-only groups. The overall recurrence rate was 12.5%, with no significant difference between the groups. Conclusions: Overall, surgical management of MRONJ resulted in favorable outcomes in a substantial proportion of patients. Within the limitations of this retrospective clinical audit, the addition of PENTO to antibiotic therapy appeared generally well tolerated, but could not result in a significant improvement in healing rates or reduction in recurrences, compared with antibiotic therapy alone, in this patient cohort. Full article

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17 pages, 2628 KB

Open AccessArticle

Probiotic and Postbiotic Interactions of Lactobacillus Strains with Candida albicans: Antifungal Effects Through Microbial Competition

by Andrea Vega-Vásconez, Diana Lucinda Castillo-Patiño, Javier Alberto Garza-Cervantes, Arlette Santacruz and José Rubén Morones-Ramírez

Antibiotics 2026, 15(3), 279; https://doi.org/10.3390/antibiotics15030279 - 10 Mar 2026

Abstract

Background: Candida albicans is the most clinically significant opportunistic fungal pathogen, and the growing resistance to conventional antifungals, particularly azoles and echinocandins, highlights the urgent need for alternative therapeutic strategies. Although lactic acid bacteria (LAB) have shown inhibitory potential against C. albicans [...] Read more.

Background: Candida albicans is the most clinically significant opportunistic fungal pathogen, and the growing resistance to conventional antifungals, particularly azoles and echinocandins, highlights the urgent need for alternative therapeutic strategies. Although lactic acid bacteria (LAB) have shown inhibitory potential against C. albicans, the relative contributions of live probiotics, heat-inactivated postbiotics, and cell-free supernatants (CFSs) have rarely been compared in parallel under physiologically relevant conditions against a clinical oral isolate. Results: This study systematically evaluated the antifungal activity of Lactiplantibacillus plantarum 299V, Lactobacillus delbrueckii subsp. bulgaricus ATCC 11842, and Lactobacillus acidophilus ATCC 4356 using co-culture assays, minimum inhibitory concentration tests, agar well diffusion assays, and optical microscopy. L. plantarum achieved the strongest inhibitory effect in co-culture, reducing C. albicans viability by 2.39 log₁₀ CFU/mL after 24 h, correlating with the greatest acidification of the culture medium. Methods: CFS from L. acidophilus inhibited fungal growth by 79.01% at native pH, declining to 28.35% upon neutralization to pH 7, confirming that antifungal efficacy is largely pH-dependent and driven by undissociated organic acids. At probiotic concentrations of 1 × 10⁹ CFU/mL, all strains completely suppressed fungal growth. Heat-inactivated postbiotics exhibited up to 95.14% inhibition in MIC assays; however, microscopic analysis revealed coaggregation between postbiotic and fungal cells, which likely interfered with optical density measurements. Conclusions: These findings establish that LAB-mediated antifungal activity is multifactorial and assay-dependent, and highlight the importance of distinguishing between probiotic, postbiotic, and CFS effects when developing LAB-based antifungal strategies. Full article

(This article belongs to the Special Issue Antibiotics: Fighting Disease and Promoting Health by Embracing the Two Sides of the Same Coin)

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16 pages, 1069 KB

Open AccessArticle

Nitric Oxide Donor Spermine-NONOate Elicits Endogenous Dispersal-Associated Transcriptional Responses to Promote Biofilm Dispersal in Pseudomonas aeruginosa

by Xavier Bertran Forga, Kathryn E. Fairfull-Smith, Jilong Qin and Makrina Totsika

Antibiotics 2026, 15(3), 278; https://doi.org/10.3390/antibiotics15030278 - 9 Mar 2026

Abstract

Background/Objectives: Bacterial biofilms are structured communities of sessile cells embedded in a self-produced extracellular matrix. Within biofilms, bacteria become highly tolerant toenvironmental stressors such as host immune responses and antimicrobial treatments. In response to specific cues, however, biofilm cells can revert to [...] Read more.

Background/Objectives: Bacterial biofilms are structured communities of sessile cells embedded in a self-produced extracellular matrix. Within biofilms, bacteria become highly tolerant toenvironmental stressors such as host immune responses and antimicrobial treatments. In response to specific cues, however, biofilm cells can revert to a planktonic free-swimming lifestyle through a process termed biofilm dispersal. When dispersed cells escape the biofilm matrix, they lose biofilm-associated antibiotic tolerance, a major barrier to treating medical biofilms. As such, dispersal-inducing compounds like nitric oxide (NO) are actively investigated as adjuvants to potentiate the biofilm-eradicating activity of existing antibiotics. We recently characterised the transcriptomic responses elicited during spontaneous biofilm dispersal in closed culture-grown Pseudomonas aeruginosa biofilms. Here, we evaluated the transcriptional profiles of P. aeruginosa biofilms treated with the NO donor Spermine-NONOate (SP-NONO) and the nitroxide C-TEMPO, an NO analogue, to determine potential pathways involved in NO-mediated dispersal. Methods: Dispersal activity on P. aeruginosa PAO1 biofilms by SP-NONOate and C-TEMPO was quantified by crystal violet staining. Cellular responses to each compound were profiled by RNA-seq on treated and untreated cells. Results: While both compounds disrupted the transcription of ANR-regulated energy metabolism pathways, only SP-NONO activated canonical NO-regulated responses. Considering that only SP-NONO showed biofilm dispersal activity in this culture system, we investigated shared transcriptional shifts in SP-NONO-treated and spontaneously dispersed biofilms to identify pathways likely involved in central dispersal responses. These mostly included genes involved in the catabolism of branched-chain amino acids (leucine, valine, isoleucine) and lysine, as well as 9 of 14 genes previously defined as transcriptional biomarkers of spontaneous biofilm dispersal. Conclusions: This study suggests that NO disrupts biofilm maturation by prematurely stimulating central pathways of spontaneous biofilm dispersal and highlights this set of biomarkers as robust indicators of dispersal responses. Full article

(This article belongs to the Section Antibiofilm Strategies)

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14 pages, 1224 KB

Open AccessArticle

Comprehensive Shotgun Metagenomic Profiling of Antibiotic Resistance Genes in Sheep and Goat Farming Environments

by Sara Gomes-Gonçalves, Jaqueline T. Bento, Guilherme Moreira, Joana Mourão, Rita Cruz, Fernando Esteves, Alexandra Lameira Baptista, Maria Aires Pereira, Pedro Caseiro, Pedro Carreira, Luís Figueira and João R. Mesquita

Antibiotics 2026, 15(3), 277; https://doi.org/10.3390/antibiotics15030277 - 9 Mar 2026

Abstract

Background: Antimicrobial resistance (AMR) is a growing global health concern, driven in part by antibiotic use in animal production systems. Despite its relevance, the microbiome and resistome of small ruminant farm environments remain largely underexplored. Methods: In this study, shotgun metagenomics was applied [...] Read more.

Background: Antimicrobial resistance (AMR) is a growing global health concern, driven in part by antibiotic use in animal production systems. Despite its relevance, the microbiome and resistome of small ruminant farm environments remain largely underexplored. Methods: In this study, shotgun metagenomics was applied to environmental samples from 46 sheep, goat and mixed-species farms across 14 municipalities in central Portugal. Results: Microbial profiling revealed a well-preserved microbiome with Pseudomonadota, Actinomycetota, Bacteroidota and Bacillota (syn. Proteobacteria, Actinobacteria, Bacteroidetes and Firmicutes respectively) as the most dominant phylum across different farm types. Regarding AMR, a total of 706 unique antimicrobial resistance genes (ARGs), covering 15 antibiotic classes, were detected. Tetracycline, aminoglycoside and macrolide resistance genes dominated across all samples, forming a conserved core resistome. While overall resistome profiles were broadly similar among farm types, significant differences were observed in specific ARG classes, such as pleuromutilin and fosfomycin. Conclusions: These findings highlight small ruminant farm environments as potential reservoirs of clinically relevant ARGs, including WHO highest priority critically important antimicrobial (HPCIA) resistance genes for macrolides (mph(c), erm(f), erm(b)) and fluoroquinolones (qnrD1), as well as critically important antimicrobial (CIA) resistance genes for glycopeptides (vanR-SC, vanR-O) and aminoglycosides (str, aadA), supporting the need to incorporate these environments into surveillance strategies. Full article

(This article belongs to the Special Issue Genomic Surveillance of Antimicrobial Resistance (AMR))

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20 pages, 1688 KB

Open AccessArticle

Overcoming Multidrug Resistance by Bacterial Efflux Pump Inhibitors in Clinical Escherichia coli Strains

by Nikoletta Szemerédi, Márta Nové, Danhui Heo, László Orosz, József Sóki and Gabriella Spengler

Antibiotics 2026, 15(3), 276; https://doi.org/10.3390/antibiotics15030276 - 9 Mar 2026

Abstract

Background/Objectives: Antimicrobial resistance (AMR) is an escalating global threat driven by antibiotic misuse and bacterial adaptation. Efflux pumps are major contributors to multidrug resistance in Escherichia coli, as they expel antibiotics and reduce their intracellular activity. This study examined efflux-mediated resistance [...] Read more.

Background/Objectives: Antimicrobial resistance (AMR) is an escalating global threat driven by antibiotic misuse and bacterial adaptation. Efflux pumps are major contributors to multidrug resistance in Escherichia coli, as they expel antibiotics and reduce their intracellular activity. This study examined efflux-mediated resistance in extended-spectrum beta-lactamase (ESBL)-producing E. coli and evaluated the potential of several efflux pump inhibitors (EPIs)—promethazine (PMZ), thioridazine (TZ), carbonyl cyanide m-chlorophenyl hydrazine (CCCP), reserpine (RES), and phenyl-arginine-β-naphthylamide (PAβN)—as therapeutic adjuncts. Methods: Antibacterial and anti-biofilm activities of EPIs were tested using broth microdilution, real-time fluorimetry, and crystal violet assays, while ceftriaxone–PMZ interactions were assessed by checkerboard analysis. Results: TZ and CCCP showed strain-specific antibacterial activity, whereas PMZ, RES, and PAβN did not exert any effect. PMZ, TZ, and especially CCCP effectively inhibited efflux pump function, while RES and PAβN were less active. Biofilm inhibition varied between strains, with PMZ and TZ producing moderate reductions. We observed a quite weak synergism between ciprofloxacin, ceftriaxone, and PMZ; however, the result was not significant. Conclusions: Overall, the results highlight the central role of efflux pumps in ESBL-producing E. coli and indicate that EPIs can reverse resistance (e.g., PMZ) and exhibit potent anti-biofilm activity and show additive interactions with antibiotics. However, further studies are needed to optimize their safety, pharmacokinetics, and antibiotic pairing for potential clinical use. Full article

(This article belongs to the Special Issue Drivers of Virulence and Antimicrobial Resistance in Emerging and Re-Emerging Pathogens)

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17 pages, 2418 KB

Open AccessArticle

Characterization of an Escherichia coli ST156 Isolate Harboring an IncHI2-Type Plasmid Co-Carrying bla_NDM-5 and mcr-1.1 Genes from Urban Wastewater Treatment Plants in Fengxian, Shanghai

by Qingyuan Zhang, Xiaohong Xie, Lixin Tao, Jian Wang, Yuan Shi, Huangfei Sheng, Chuanlong Liu, Hongwei Zhao, Meihua Liu and Jun Feng

Antibiotics 2026, 15(3), 275; https://doi.org/10.3390/antibiotics15030275 - 6 Mar 2026

Abstract

Background: The emergence of carbapenem-resistant enterobacteriaceae (CRE) co-harboring the mcr-1.1 gene and carbapenemase-encoding genes poses a severe threat to public health. Urban wastewater treatment plants (WWTPs) act as natural reservoirs and hotspots for the dissemination of antimicrobial resistance genes (ARGs). This study [...] Read more.

Background: The emergence of carbapenem-resistant enterobacteriaceae (CRE) co-harboring the mcr-1.1 gene and carbapenemase-encoding genes poses a severe threat to public health. Urban wastewater treatment plants (WWTPs) act as natural reservoirs and hotspots for the dissemination of antimicrobial resistance genes (ARGs). This study aimed to elucidate the molecular characteristics of CRE carrying mcr-1.1 in urban WWTPs. Methods: Samples were collected from the influent of urban WWTPs in Fengxian, Shanghai, from April 2024 to March 2025. mcr-1.1-positive Escherichia coli (E. coli) isolates were screened using real-time PCR, and their antimicrobial susceptibility was determined via the broth microdilution method. Plasmid conjugation assays were performed with E. coli C600 as the recipient strain. Whole-genome sequencing (WGS) was carried out to analyze the molecular characteristics of mcr-1.1-positive E. coli isolates. Results: A total of 312 samples were collected, and 5 (1.6%) mcr-1.1-positive E. coli isolates were identified. All isolates were multidrug-resistant (MDR) but susceptible to tigecycline (TIG). WGS of strain EC0176 (sequence type 156 [ST156], enteroaggregative E. coli [EAEC]) detected the presence of bla_NDM-5, bla_TEM-1, bla_CTX-M-55, and mcr-1.1 as well as related virulence genes. Further analysis revealed that pEC0176 was an IncHI2-type plasmid co-harboring mcr-1.1, bla_NDM-5, arr-3, aph(4)-Ia, aph(3′)-Ia, aac(3)-IVa, and mph(A). The plasmid pEC0176 harbored similar backbones as p20014-MCR, p2017.03.02CC_1, pSC2017167-mcr-256k, pEC17CM13_MCR and pGDE043-mcr1, including the type IV secretion system (T4SS) and IncHI-type conjugal transfer genes. Conjugation experiments confirmed that pEC0176 could be horizontally transferred into E. coli C600, with an average transfer efficiency of 3.3 × 10⁻². Phylogenetic analysis showed that the MCR-1 protein of EC0176 is closely related to that of two human-derived E. coli strains from China (GenBank accession: AVR64822.1 and WP_076611062.1). Conclusions: To our knowledge, this is the first report of E. coli ST156 carrying an IncHI2-type plasmid co-harboring mcr-1.1 and bla_NDM-5 from urban WWTPs in Fengxian, Shanghai. Our findings underscore the severe status of bacterial antimicrobial resistance and emphasize the necessity of enhancing antimicrobial resistance surveillance in urban WWTPs. Full article

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