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Keywords = AmpC beta-lactamase

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17 pages, 325 KB  
Article
Prevalence and Antimicrobial Resistance Profiles of E. coli, P. mirabilis, and E. cloacae Complex Isolated from Dogs with Otitis Externa
by Ionela Popa, Ionica Iancu, Alexandru Gligor, Kalman Imre, Emil Tîrziu, Timea Bochiș, Călin Pop, Janos Degi, Andrei Ivan, Michael Dahma, Ana-Maria Plotuna, Sebastian Alexandru Popa, Marius Pentea, Viorel Herman and Ileana Nichita
Antibiotics 2026, 15(4), 343; https://doi.org/10.3390/antibiotics15040343 - 27 Mar 2026
Cited by 1 | Viewed by 935
Abstract
Background/Objectives: Antimicrobial resistance (AMR) in companion animals is an emerging public health threat due to zoonotic potential and limited therapeutic options. Dogs with otitis externa may harbor multidrug-resistant (MDR) bacteria, including Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), [...] Read more.
Background/Objectives: Antimicrobial resistance (AMR) in companion animals is an emerging public health threat due to zoonotic potential and limited therapeutic options. Dogs with otitis externa may harbor multidrug-resistant (MDR) bacteria, including Escherichia coli (E. coli), Proteus mirabilis (P. mirabilis), and Enterobacter cloacae complex (E. cloacae complex), some producing extended-spectrum beta-lactamase (ESBL) or AmpC β-lactamases. This study aimed to assess the prevalence, AMR patterns, MDR occurrence, β-lactamase production, and co-infection profiles of these pathogens in canine otitis externa. Methods: Ear canal samples were collected from 592 dogs presenting clinical signs of otitis externa, with one sample per dog included in the analysis. Samples were collected from veterinary clinics in Timiș County, Romania, from 2022 to 2025. Samples were cultured on blood agar and MacConkey agar, followed by biochemical testing and MALDI-TOF mass spectrometry for bacterial identification. Antimicrobial susceptibility testing against 15 agents across six classes was performed using the VITEK® 2 system. MDR and β-lactamase production (ESBL, AmpC) were determined according to CLSI 2018 veterinary guidelines. Co-isolation with bacterial and fungal species were recorded. Results: E. coli, P. mirabilis, and E. cloacae complex were isolated in 9.12%, 6.25%, and 1.2% of cases, respectively. E. coli exhibited the highest resistance to aminoglycosides (tobramycin 72.2%, gentamicin 61.1%) and full susceptibility to carbapenems. P. mirabilis showed the highest resistance to ampicillin (54%) and trimethoprim + sulfamethoxazole (46%), with complete susceptibility to carbapenems and fluoroquinolones. E. cloacae complex displayed universal resistance to cephalosporins but remained susceptible to non-cephalosporin β-lactams (piperacillin–tazobactam), carbapenems and aminoglycosides. MDR prevalence was 35.2% for E. coli, 18.9% for P. mirabilis, and 14.3% for the E. cloacae complex. ESBL production was detected in 13% of E. coli and 8.1% of P. mirabilis isolates, while all E. cloacae complex isolates were AmpC-positive. Co-isolations were common, primarily involving Staphylococcus pseudintermedius (S. pseudintermedius) and Malassezia pachydermatis (M. pachydermatis). Conclusions: MDR and β-lactamase-producing bacteria were identified in dogs with otitis externa, emphasizing the importance of routine antimicrobial susceptibility testing, targeted therapy based on local resistance profiles, and continuous AMR surveillance to prevent treatment failure and mitigate zoonotic risk. Full article
42 pages, 3257 KB  
Systematic Review
Antimicrobial Resistance in Selected Enterobacteriaceae from Broilers and Their Environment: ESBL, AmpC, Carbapenemases, Colistin, and Fluoroquinolone Resistance—A Systematic Review and Meta-Analysis
by Julia von Kiparski, Nunzio Sarnino, Diana Vargas, Aleksandra Atanasova and Roswitha Merle
Antibiotics 2025, 14(12), 1268; https://doi.org/10.3390/antibiotics14121268 - 15 Dec 2025
Cited by 2 | Viewed by 1487
Abstract
Background/Objectives: Antimicrobial resistance (AMR) threatens global public health. This systematic review and meta-analysis, as part of the “ENVIRE” project (interventions to control the dynamics of antimicrobial resistance from chickens through the environment), assesses the prevalence of phenotypic and genotypic resistance, including extended-spectrum beta-lactamases [...] Read more.
Background/Objectives: Antimicrobial resistance (AMR) threatens global public health. This systematic review and meta-analysis, as part of the “ENVIRE” project (interventions to control the dynamics of antimicrobial resistance from chickens through the environment), assesses the prevalence of phenotypic and genotypic resistance, including extended-spectrum beta-lactamases (ESBLs), AmpC beta-lactamases, carbapenemases, colistin, and fluoroquinolone resistance, in broiler chickens and their environment. Methods: The analysis covers the years 2002–2022, focusing on Escherichia (E.) coli, Klebsiella spp., Enterobacter spp., and Citrobacter spp. in fecal, meat, environmental, and other-than-feces samples from observational studies published in PubMed and Web of Science. Quality assessment was performed using the Alberta Heritage Foundation criteria. Results: Data from 170 studies, conducted in Europe, North Africa, and North America, were included. The most frequently studied resistance was to beta-lactam, with focus on ESBL-producing and AmpC beta-lactamase isolates. The pooled prevalence of ESBL-resistant E. coli observed in meat samples at 41% and in fecal samples at 38% demonstrated significant heterogeneity between the studies. The negative binomial regression analysis of prevalence data revealed significantly higher ESBL-producing E. coli rates in European meat samples compared to North African samples. Conclusions: This systematic review revealed substantial variation in prevalence and emphasizes the need for standardized surveillance systems and robust study designs. Full article
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24 pages, 17742 KB  
Article
The Rapid CarbaLux Combination Test to Uncover Bacterial Resistance and Heteroresistance Prior to Antibiotic Treatment
by Hans Rudolf Pfaendler and Hans-Ulrich Schmidt
Diagnostics 2025, 15(20), 2624; https://doi.org/10.3390/diagnostics15202624 - 17 Oct 2025
Viewed by 1209
Abstract
Background/Objectives: In this proof-of-concept study, the objective was to evaluate the phenotypic CarbaLux combination rapid test in terms of guiding the therapy of infections caused by multidrug-resistant Gram-negative bacteria with carbapenemase inhibitors and carbapenems, and to compare its results and practicability with standard [...] Read more.
Background/Objectives: In this proof-of-concept study, the objective was to evaluate the phenotypic CarbaLux combination rapid test in terms of guiding the therapy of infections caused by multidrug-resistant Gram-negative bacteria with carbapenemase inhibitors and carbapenems, and to compare its results and practicability with standard diagnostic methods. Methods: In the classical CarbaLux test, a fluorescent carbapenem serves as a UV–visible diagnostic surrogate for clinically used carbapenem antibiotics. When exposed to extracted carbapenemases from bacterial colony growth on agar plates, fluorescence rapidly disappears, showing whether monotherapy with carbapenems is possible or must be rejected. It was expected that a specific inhibitor that protects imipenem or meropenem from enzymatic deactivation during antibacterial therapy would perform the same in vitro with fluorescent carbapenem and preserve its fluorescence. The new additional CarbaLux combination test is used if the classic test is positive for carbapenemases: a classic test tube pre-dosed with fluorescent carbapenem is spiked with cloxacillin; with recently launched carbapenemase inhibitors, e.g., avibactam, relebactam, zidebactam, nacubactam, or vaborbactam; or with picolinic acid. Fourteen Enterobacterales and six Acinetobacter baumannii isolates were analyzed. Results: At fixed concentrations, the new inhibitors protected fluorescent carbapenem from bacterial KPC-mediated inactivation and partially from AmpC beta-lactamase-mediated inactivation. In addition, avibactam also effectively inhibited OXA-48-like enzymes. Cloxacillin selectively inhibited AmpC beta-lactamases extracted from Enterobacter complex species. Non-therapeutic picolinic acid was specific for metallo-beta-lactamases and thus identified infections by pathogens that cannot be treated with carbapenems alone or in combination. Conclusions: Inhibitor/fluorescent carbapenem mixtures corresponding to therapeutic inhibitor/carbapenem combinations allow us to visualize the efficacy of carbapenemase inhibitors. The in vitro results are consistent with clinical experience regarding combination therapy. Enzymatic assays provide a rapid yes/no answer for carbapenem mono- or combination therapy and offer several advantages over current carbapenemase testing methods. In contrast to PCR and lateral flow tests, which only target a selection of carbapenemases, enzymatic assays work by employing a reproducible phenotypic mechanism. They are simpler, broader in scope, and more cost-effective; they can also detect antimicrobial heteroresistance or AmpC beta-lactamase hyperproduction, which is normally undetected when performing automated antibiotic susceptibility testing. The new tests are suitable for clinical diagnosis, public health purposes, and infection control. Full article
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16 pages, 385 KB  
Article
Extended-Spectrum β-Lactamase-/AmpC-Producing Escherichia coli and Associated Risk Factors in Shelter Dogs: A Baseline Study in North Macedonia
by Ivana Shikoska, Sanja Duvnjak, Tom Koritnik, Bojana Chapkunovska, Jane Vlahov, Marija Ratkova Manovska, Aleksandar Cvetkovikj and Iskra Cvetkovikj
Microbiol. Res. 2025, 16(9), 206; https://doi.org/10.3390/microbiolres16090206 - 17 Sep 2025
Viewed by 1527
Abstract
Antimicrobial resistance (AMR) is a significant public health concern in companion animals, yet systematic surveillance in North Macedonia is lacking. This study investigated the prevalence of resistance in Escherichia coli isolated from 112 fecal samples from dogs in six shelters in North Macedonia [...] Read more.
Antimicrobial resistance (AMR) is a significant public health concern in companion animals, yet systematic surveillance in North Macedonia is lacking. This study investigated the prevalence of resistance in Escherichia coli isolated from 112 fecal samples from dogs in six shelters in North Macedonia and evaluated the associated risk factors, providing the first baseline dataset for this population. High resistance was observed for sulfamethoxazole (68.75%), ampicillin (52.68%), and ciprofloxacin (41.07%). Multidrug resistance was present in 50% of the isolates, with 17 (15.17%) confirmed as ESBL producers. Additionally, 18 isolates (16.1%) were identified as AmpC producers, 16 of which carried the blaCMY-2 gene. Notably, 72.2% of ESBL/AmpC isolates were resistant to ertapenem despite the absence of carbapenemase genes, a finding that warrants further investigation. Risk factors such as shared housing, longer shelter stays, and frequent empirical antimicrobial use were identified as probable contributors to the carriage of ESBL-/AmpC-producing E. coli. None of the shelters had antimicrobial stewardship protocols or routine diagnostic testing, revealing critical gaps in infection control and antimicrobial practices. These findings underscore the urgent need for antimicrobial stewardship and surveillance in North Macedonia’s companion animal populations within the One Health framework. Full article
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24 pages, 3027 KB  
Article
Resisting the Final Line: Phenotypic Detection of Resistance to Last-Resort Antimicrobials in Gram-Negative Bacteria Isolated from Wild Birds in Northern Italy
by Maria Cristina Rapi, Joel Filipe, Laura Filippone Pavesi, Stefano Raimondi, Maria Filippa Addis, Maria Pia Franciosini and Guido Grilli
Animals 2025, 15(15), 2289; https://doi.org/10.3390/ani15152289 - 5 Aug 2025
Cited by 2 | Viewed by 1943
Abstract
Antimicrobial resistance (AMR) is a growing global health threat, with wild birds increasingly recognized as potential reservoirs of resistant pathogens and as sentinels of environmental AMR. This study investigated the occurrence and AMR profiles of Gram-negative bacteria isolated from wild birds that died [...] Read more.
Antimicrobial resistance (AMR) is a growing global health threat, with wild birds increasingly recognized as potential reservoirs of resistant pathogens and as sentinels of environmental AMR. This study investigated the occurrence and AMR profiles of Gram-negative bacteria isolated from wild birds that died at the Wildlife Rescue Center in Vanzago, Lombardy, in 2024. Cloacal swabs were collected from 112 birds representing various ecological categories. A total of 157 Gram-negative bacteria were isolated and identified, including clinically relevant genera and species, such as Escherichia coli, Klebsiella pneumoniae, Enterobacter spp., Salmonella spp., Pseudomonas aeruginosa, and Acinetobacter baumannii. Antimicrobial susceptibility testing revealed resistance to first-line and critically important antimicrobials, including those exclusively authorized for human use. Notably, a phenotype compatible with Extended-Spectrum Beta-Lactamase (ESBL) production was detected in four out of ten (40%) K. pneumoniae isolates. In addition, 20 out of the 157 (12.7%) isolated bacteria phenotypically exhibited a resistance profile indicative of AmpC beta-lactamase (AmpC) production, including Enterobacter spp. and P. aeruginosa. Resistance patterns were particularly interesting in birds with carnivorous, scavenging, or migratory-associated behaviors. These findings highlight the role of wild birds in the ecology and dissemination of antimicrobial-resistant bacteria (ARB) and highlight the need for wildlife-based AMR monitoring programs as part of a One Health approach. Full article
(This article belongs to the Section Birds)
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9 pages, 234 KB  
Article
Should Cefoxitin Non-Susceptibility in Ceftriaxone-Susceptible E. coli and K. pneumoniae Prompt Concerns Regarding Plasmid-Mediated AmpC Resistance? A Genomic Characterization and Summary of Treatment Challenges in Singapore
by Jonathan Jinpeng Foo, Ying Ying Ong, Clement Kin Ming Tsui, David C. Lye, De Partha Pratim, Nurhidayah Binte Mohamed Yazid, Swaine L. Chen, Shawn Vasoo and Tat Ming Ng
Antibiotics 2025, 14(7), 722; https://doi.org/10.3390/antibiotics14070722 - 18 Jul 2025
Viewed by 2194
Abstract
Objectives: Plasmid-mediated AmpC beta-lactamases represent a growing clinical concern in Enterobacterales, with challenges in diagnostic approaches, limited data on clinical outcomes, and our incomplete understanding of their regulatory mechanisms warranting the need for further investigation. Methods: This retrospective study examined the genomic [...] Read more.
Objectives: Plasmid-mediated AmpC beta-lactamases represent a growing clinical concern in Enterobacterales, with challenges in diagnostic approaches, limited data on clinical outcomes, and our incomplete understanding of their regulatory mechanisms warranting the need for further investigation. Methods: This retrospective study examined the genomic and clinical characteristics of cefoxitin-non-susceptible, ceftriaxone-susceptible Escherichia coli and Klebsiella pneumoniae bloodstream isolates collected from a tertiary hospital in Singapore. Whole-genome sequencing was performed to detect ampC genes, subtypes, and associated regulatory elements. Results: Among 108 cefoxitin-non-susceptible isolates, only 15 (13.9%) harboured plasmid-mediated ampC, suggesting that cefoxitin non-susceptibility alone in ceftriaxone susceptible isolates was not predictive of ampC carriage. All plasmid-ampC isolates were from the blaDHA-1 subtype and carried ampR, a known transcriptional regulator of inducible beta-lactamase expression. Notably, five non-ampC carrying Klebsiella isolates displayed truncations in ompK35 and ompK36, which could potentially contribute to reduced cefoxitin susceptibility via porin loss. Conclusions: These findings underscore the limited diagnostic utility of cefoxitin susceptibility testing for detecting plasmid-mediated ampC producers and highlight the clinical relevance of regulatory genes such as ampR in mediating inducible resistance. The routine incorporation of molecular diagnostics or genome sequencing may be necessary to improve detection accuracy and inform antimicrobial stewardship strategies. Full article
19 pages, 1281 KB  
Article
Phenotypic and Genotypic Characterization of ESBL and AmpC β-Lactamase-Producing E. coli Isolates from Poultry in Northwestern Romania
by Anca Rus, Iulia-Maria Bucur, Kalman Imre, Andreea Talida Tirziu, Andrei Alexandru Ivan, Radu Valentin Gros, Alex Cristian Moza, Sebastian Alexandru Popa, Alexandra Ban-Cucerzan and Emil Tirziu
Antibiotics 2025, 14(6), 578; https://doi.org/10.3390/antibiotics14060578 - 5 Jun 2025
Cited by 7 | Viewed by 2266
Abstract
Background/Objectives: The widespread use of antibiotics in animal husbandry has led to an increase in antimicrobial-resistant Escherichia coli, particularly strains producing extended-spectrum β-lactamases (ESBL) and AmpC β-lactamases. This study aimed to isolate and characterize such strains from fecal samples of broiler [...] Read more.
Background/Objectives: The widespread use of antibiotics in animal husbandry has led to an increase in antimicrobial-resistant Escherichia coli, particularly strains producing extended-spectrum β-lactamases (ESBL) and AmpC β-lactamases. This study aimed to isolate and characterize such strains from fecal samples of broiler chickens (n = 71) and slaughtered turkeys (n = 31) in northwestern Romania. Methods: Antimicrobial susceptibility testing and PCR were used to evaluate phenotypic resistance patterns and detect the presence of resistance genes (AmpC, blaZ, and blaTEM). Results: The results showed that 55% of turkey and 61% of broiler isolates were presumptive ESBL/AmpC producers. Among all isolates, 50% were classified as extensively drug-resistant (XDR), 44% as multidrug-resistant (MDR), and only 6% were fully susceptible. Gene detection revealed an overall prevalence of 44.2% for AmpC, 72.7% for blaZ, and 58.1% for blaTEM, yielding a total penetrance of 51.09%. The diagnostic odds ratio (DOR) values, ranging from 0.67 to 81, suggest the efficacy of the antibiotic susceptibility testing method used in detecting the presence of these resistance genes. Conclusion: Overall, these findings highlight a significant burden of antimicrobial-resistant, poultry-associated E. coli strains, warranting stricter antimicrobial stewardship. Full article
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16 pages, 1028 KB  
Review
Characterization of Antibiotic Resistance in Shewanella Species: An Emerging Pathogen in Clinical and Environmental Settings
by Shahid Sher, Gary P. Richards, Salina Parveen and Henry N. Williams
Microorganisms 2025, 13(5), 1115; https://doi.org/10.3390/microorganisms13051115 - 13 May 2025
Cited by 12 | Viewed by 4624
Abstract
Antibiotic resistance is increasing at an alarming rate worldwide, in large part due to their misuse and improper disposal. Antibiotics administered to treat human and animal diseases, including feed supplements for the treatment or prevention of disease in farm animals, have contributed greatly [...] Read more.
Antibiotic resistance is increasing at an alarming rate worldwide, in large part due to their misuse and improper disposal. Antibiotics administered to treat human and animal diseases, including feed supplements for the treatment or prevention of disease in farm animals, have contributed greatly to the emergence of a multitude of antibiotic-resistant pathogens. Shewanella is one of many bacteria that have developed antibiotic resistance, and in some species, multiple-antibiotic resistance (MAR). Shewanella is a rod-shaped, Gram-negative, oxidase-positive, and H2S-producing bacterium that is naturally found in the marine environment. In humans, Shewanella spp. can cause skin and soft tissue infections, septicemia, cellulitis, osteomyelitis, and ear and wound infections. Some Shewanella have been shown to be resistant to a variety of antibiotics, including beta-lactams, aminoglycoside, quinolones, third- or fourth-generation cephalosporins, and carbapenems, due to the presence of genes such as the blaOXA-class D beta-lactamase-encoding gene, blaAmpC-class-C beta-lactamase-encoding gene, and the qnr gene. Bacteria can acquire and transmit these genes through different horizontal gene-transmission mechanisms such as transformation, transduction, and conjugation. The genes for antibiotic resistance are present on Shewanella chromosomes and plasmids. Apart from this, heavy metals such as arsenic, mercury, cadmium, and chromium can also increase antibiotic resistance in Shewanella due to co-selection processes such as co-resistance, cross resistance, and co-regulation mechanisms. Antibiotics and drugs enter Shewanella spp. through pores or gates in their cell wall and may be ejected from the bacteria by efflux pumps, which are the first line of bacterial defense against antibiotics. Multiple-drug resistant Shewanella can be particularly difficult to control. This review focuses on the phenotypic and genomic characteristics of Shewanella that are involved in the increase in antimicrobial resistance in this bacterium. Full article
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18 pages, 599 KB  
Article
Mapping Antimicrobial Resistance in Escherichia coli and Klebsiella pneumoniae from Complicated Urinary Tract Infections in Oman: Phenotypic and Genotypic Insights
by Nawal AL Shizawi, Zaaima AL Jabri, Fatima Khan, Hiba Sami, Turkiya AL Siyabi, Zakariya AL Muharrmi, Srinivasa Rao Sirasanagandla and Meher Rizvi
Diagnostics 2025, 15(9), 1062; https://doi.org/10.3390/diagnostics15091062 - 22 Apr 2025
Cited by 4 | Viewed by 3621
Abstract
Background: Mapping the local etiology and susceptibility of common pathogens causing complicated urinary tract infection (cUTI) is important for promoting evidence-based antimicrobial prescribing. Evaluating the prevalence of extended-spectrum beta-lactamase (ESBL), AmpC beta-lactamase (AmpC), and carbapenemase-producing Enterobacterales (CPEs) is equally important as it informs [...] Read more.
Background: Mapping the local etiology and susceptibility of common pathogens causing complicated urinary tract infection (cUTI) is important for promoting evidence-based antimicrobial prescribing. Evaluating the prevalence of extended-spectrum beta-lactamase (ESBL), AmpC beta-lactamase (AmpC), and carbapenemase-producing Enterobacterales (CPEs) is equally important as it informs treatment guidelines and empiric management. Whole genome sequencing (WGS) enhances antimicrobial resistance (AMR) surveillance by complementing phenotypic antimicrobial susceptibility testing, offering deeper insights into resistance mechanisms, transmissions, and evolutions. Integrating it into routine AMR monitoring can significantly improve global efforts to combat antimicrobial resistance. Methods: Antimicrobial susceptibility profiles of isolates from cUTI were collected from patients presenting with Sultan Qaboos University Hospital, Muscat and Suhar Hospital, Suhar, Oman. Automated systems as well as manual methods were used for detection of ESBL, AmpC, and CPE. ESBLs, AmpC β-lactamases, and CPEs were further detected by manual methods: double-disk synergy test for ESBL; disk approximation assay and D69C AmpC detection set for AmpC, and mCIM and KPC/IMP/NDM/VIM/OXA-48 Combo test kit for CPE. WGS was carried out in 11 FOX-resistant E. coli and (22 carbapenem-resistant K. pneumoniae) isolates with varying susceptibilities to identify circulating clades, AMR genes, and plasmids. Bioinformatic analysis was performed using online tools. Results: The susceptibility patterns of E. coli from cUTI were as follows: nitrofurantoin (96%), fosfomycin (100%), fluoroquinolones (44%), aminoglycosides (93%), piperacillin-tazobactam (95%), and carbapenems (98%). In comparison, susceptibility rates of K. pneumoniae were far lower: nitrofurantoin (38%), fosfomycin (89%), aminoglycosides (82%), piperacillin-tazobactam (72%), and carbapenems (83%). K. pneumoniae, however, was more susceptible to fluoroquinolones at 47% in comparison to E. coli. The prevalence of ESBL among E. coli and K. pneumoniae was 37.2% and CRE was 6.2% while the estimated prevalence of AmpC was 5.4%. It was observed that E. coli was the predominant ESBL and AmpC producer, while K. pneumoniae was the major carbapenem-resistant Enterobacterales (CREs) producer. No predominant multi-locus sequence typing (MLST) lineage was observed in AmpC-producing E. coli with nine E. coli MLST lineages being identified from eleven isolates: ST-10, ST-69, ST-77, ST-131, ST-156, ST-167, ST-361, ST-1125, and ST-2520. On the other hand, a less diverse MLST spectrum (ST-2096, ST-231, ST-147, ST-1770, and ST-111) was observed in the CRE K. pneumoniae. Among the five MLST lineages, ST-2096 (twelve isolates) and ST-147 (seven isolates) predominated. WGS revealed that DHA-1 was the predominant plasmid-mediated AmpC gene in E. coli, while OXA-232 and NDM-5 were the most common carbapenemase genes in K. pneumoniae. All E. coli DHA-1-positive isolates co-harbored the quinolone resistance gene qnrB4 and the sulfonamide resistance gene sul1 while no aminoglycoside resistance genes were detected. The majority of CPE CRE K. pneumoniae carried other β-lactamase genes, such as blaCTX-M-15, blaSHV, and blaTEM; all co-harbored the quinolone resistance gene OqxAB; and 77% carried the aminoglycoside resistance gene armA. Conclusions: Our results suggest that fosfomycin is an excellent empiric choice for treating complicated cystitis caused by both E. coli and K. pneumoniae, while nitrofurantoin is an appropriate choice for E. coli cystitis but not for K. pneumoniae. Aminoglycosides and piperacillin-tazobactam are excellent intravenous alternatives that spare carbapenems. DHA-1 was the predominant AmpC in E. coli, while OXA-232 and NDM-5 were the predominant carbapenemases in K. pneumoniae. In AmpC-producing E. coli, no MLST predominated, suggesting a significant flux in E. coli with lack of stable clades in this region. In contrast, ST-2096 and ST-147 predominated in CRE Klebsiella pneumoniae, suggesting a stable circulation of these in Oman. WGS profiling provides a deeper understanding of the genetic basis of resistance and enhances surveillance and offers comprehensive insights into pathogen evolution and transmission patterns. Full article
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20 pages, 12743 KB  
Article
Aeromonas Species Diversity, Virulence Characteristics, and Antimicrobial Susceptibility Patterns in Village Freshwater Aquaculture Ponds in North India
by Alka Nokhwal, Rajesh Kumar Vaid, Taruna Anand, Ravikant Verma and Rachna Gulati
Antibiotics 2025, 14(3), 294; https://doi.org/10.3390/antibiotics14030294 - 12 Mar 2025
Cited by 7 | Viewed by 4203
Abstract
Background/Objectives: Motile aeromonads are ubiquitous aquatic Gram-negative opportunistic pathogens with environmental, animal, aquatic, and human health implications. Methods: Motile aeromonads were isolated from village pond water samples (n = 100) of the Hisar district of Haryana state in India. Selective isolation and [...] Read more.
Background/Objectives: Motile aeromonads are ubiquitous aquatic Gram-negative opportunistic pathogens with environmental, animal, aquatic, and human health implications. Methods: Motile aeromonads were isolated from village pond water samples (n = 100) of the Hisar district of Haryana state in India. Selective isolation and enumeration were followed by biochemical and genotypic identification using gyrB gene; evaluation of seven putative virulence factors and antimicrobial resistance studies and determination of extended spectrum beta lactamase (ESBL) and AmpC beta lactamase (ACBL) enzyme-producing abilities took place. Results: The viable counts of motile aeromonads varied from 1.6 × 102 CFU/mL to 1.2 × 108 CFU/mL. Six species of Aeromonas were identified with high prevalence of A. veronii (74.7%), followed by A. caviae (8.9%), A. hydrophila (7.6), A. jandaei (5%), A. sobria (2.5%), and A. dhakensis (1.3%). PCR amplification of seven genes related to virulence indicated that the majority of the isolates were positive for enolase (eno, 98%), cytotoxic enterotoxin (act, 88%), and hemolysin (asa1, 86%). Many isolates were also positive for type III secretion system inner membrane component (ascV, 53%), ADP-ribosylating toxin (aexT, 47%), and extracellular hemolysin (ahh1, 4%). The antimicrobial resistance (AMR) profile of the isolated Aeromonas isolates indicated the high resistance observed to nalidixic acid (40.2%), cefoxitin (33%), and imipenem (6.2%). In addition, the occurrence of 10.3% ESBL, 32% ACBL, and 29.9% multi-drug resistant (MDR) isolates is alarming. Phylogenetic analysis of gyrB sequences of A. veronii isolates (n = 59) together with GenBank sequences of A. veronii from different geographical regions of the world indicated high genotypic diversity. Conclusions: the village aquaculture ponds in Hisar district have a high occurrence of MDR A. veronii, A. hydrophila, and A. caviae, posing significant animal and public health concern. Full article
(This article belongs to the Section Antibiotics in Animal Health)
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9 pages, 623 KB  
Communication
Phenotypic Ultra-Rapid Antimicrobial Susceptibility Testing for Ceftazidime–Avibactam: In Support of Antimicrobial Stewardship
by Inês Martins-Oliveira, Blanca Pérez-Viso, Rosário Gomes, David Abreu, Ana Silva-Dias, Rafael Cantón and Cidália Pina-Vaz
Microorganisms 2025, 13(2), 414; https://doi.org/10.3390/microorganisms13020414 - 13 Feb 2025
Viewed by 2040
Abstract
Ceftazidime–avibactam (CZA) is a potent broad-spectrum drug combination covering extended-spectrum β-lactamases, AmpC, and carbapenemases of class A and D, OXA-48-type producers. Rapid antimicrobial susceptibility testing is crucial for the timely de-escalation/escalation of therapy. We evaluate CZA susceptibility using the CE-IVD FASTgramneg kit (FASTinov [...] Read more.
Ceftazidime–avibactam (CZA) is a potent broad-spectrum drug combination covering extended-spectrum β-lactamases, AmpC, and carbapenemases of class A and D, OXA-48-type producers. Rapid antimicrobial susceptibility testing is crucial for the timely de-escalation/escalation of therapy. We evaluate CZA susceptibility using the CE-IVD FASTgramneg kit (FASTinov®), a ground-breaking 2 h assay, based on flow cytometry technology for antimicrobial susceptibility testing. The assay involved rapid bacterial extraction and purification from positive blood cultures (PBCs), followed by a 1 h 37 °C incubation and flow cytometry analysis (Cytoflex, Beckman-Coulter). The susceptibility report was generated using a proprietary software and interpreted using EUCAST and CLSI 2024 criteria. Sensitivity and specificity were calculated against a reference standardized method (disk diffusion) according to ISO20776-2:2021. Overall, 135 Enterobacterales and 73 Pseudomonas aeruginosa isolates were studied. Thirty-four isolates were resistant to CZA, including six P. aeruginosa and 28 Enterobacterales (24 metallo-beta-lactamase producers, three KPC variants, and one co-producing KPC+NDM). Sensitivity and specificity reached 100% when using EUCAST and CLSI criteria compared with the reference method. The FASTinov ultra-rapid susceptibility assay for CZA demonstrated excellent results, potentially enabling de-escalation/escalation even before the second dose. Combining the speed of a molecular assay with the comprehensive information of a phenotypic test offers valuable insights for treatment decisions. Full article
(This article belongs to the Section Antimicrobial Agents and Resistance)
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17 pages, 571 KB  
Review
Antibiotic Treatment of Infections Caused by AmpC-Producing Enterobacterales
by Gianpiero Tebano, Irene Zaghi, Monica Cricca and Francesco Cristini
Pharmacy 2024, 12(5), 142; https://doi.org/10.3390/pharmacy12050142 - 21 Sep 2024
Cited by 23 | Viewed by 24511
Abstract
AmpC enzymes are a class of beta-lactamases produced by Gram-negative bacteria, including several Enterobacterales. When produced in sufficient amounts, AmpCs can hydrolyze third-generation cephalosporins (3GCs) and piperacillin/tazobactam, causing resistance. In Enterobacterales, the AmpC gene can be chromosomal- or plasmid-encoded. Some species, particularly Enterobacter [...] Read more.
AmpC enzymes are a class of beta-lactamases produced by Gram-negative bacteria, including several Enterobacterales. When produced in sufficient amounts, AmpCs can hydrolyze third-generation cephalosporins (3GCs) and piperacillin/tazobactam, causing resistance. In Enterobacterales, the AmpC gene can be chromosomal- or plasmid-encoded. Some species, particularly Enterobacter cloacae complex, Klebsiella aerogenes, and Citrobacter freundii, harbor an inducible chromosomal AmpC gene. The expression of this gene can be derepressed during treatment with a beta-lactam, leading to AmpC overproduction and the consequent emergence of resistance to 3GCs and piperacillin/tazobactam during treatment. Because of this phenomenon, the use of carbapenems or cefepime is considered a safer option when treating these pathogens. However, many areas of uncertainty persist, including the risk of derepression related to each beta-lactam; the role of piperacillin/tazobactam compared to cefepime; the best option for severe or difficult-to-treat cases, such as high-inoculum infections (e.g., ventilator-associated pneumonia and undrainable abscesses); the role of de-escalation once clinical stability is obtained; and the best treatment for species with a lower risk of derepression during treatment (e.g., Serratia marcescens and Morganella morganii). The aim of this review is to collate the most relevant information about the microbiological properties of and therapeutic approach to AmpC-producing Enterobacterales in order to inform daily clinical practice. Full article
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11 pages, 1597 KB  
Article
Genomic Analysis of Enterobacter Species Isolated from Patients in United States Hospitals
by Fred C. Tenover and Isabella A. Tickler
Antibiotics 2024, 13(9), 865; https://doi.org/10.3390/antibiotics13090865 - 10 Sep 2024
Cited by 4 | Viewed by 2914
Abstract
We analyzed the whole genome sequences (WGS) and antibiograms of 35 Enterobacter isolates, including E. hormaechei and E. asburiae, and the recently described E. bugandensis, E. kobei, E. ludwigii, and E. roggenkampii species. Isolates were obtained from human blood [...] Read more.
We analyzed the whole genome sequences (WGS) and antibiograms of 35 Enterobacter isolates, including E. hormaechei and E. asburiae, and the recently described E. bugandensis, E. kobei, E. ludwigii, and E. roggenkampii species. Isolates were obtained from human blood and urinary tract infections in patients in the United States. Our goal was to understand the genetic diversity of antimicrobial resistance genes and virulence factors among the various species. Thirty-four of 35 isolates contained an AmpC class blaACT allele; however, the E. roggenkampii isolate contained blaMIR-5. Of the six Enterobacter isolates resistant to ertapenem, imipenem, and meropenem, four harbored a carbapenemase gene, including blaKPC or blaNDM. All four isolates were mCIM-positive. The remaining two isolates had alterations in ompC genes that may have contributed to the resistance phenotype. Interpretations of cefepime test results were variable when disk diffusion and automated broth microdilution results were compared due to the Clinical Laboratory and Standards Institute use of the “susceptible dose-dependent” classification. The diversity of the blaACT alleles paralleled species identifications, as did the presence of various virulence genes. The classification of recently described Enterobacter species is consistent with their resistance gene and virulence gene profiles. Full article
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15 pages, 769 KB  
Article
Extended-Spectrum Beta-Lactamase- and Plasmidic AmpC-Producing Enterobacterales among the Faecal Samples in the Bulgarian Community
by Petya Stankova, Lyudmila Boyanova, Daniela Atanasova, Sashka Mihaylova, Mariya Sredkova, Raina Gergova, Kalina Mihova and Rumyana Markovska
Microorganisms 2024, 12(9), 1777; https://doi.org/10.3390/microorganisms12091777 - 28 Aug 2024
Cited by 2 | Viewed by 2492
Abstract
The aim of the present work was to genetically characterise cefotaxime-resistant enterobacteria isolated from community carriers in Bulgaria. In total, 717 faecal samples from children and adults in five medical centres in Sofia, Pleven and Burgas were examined. Antimicrobial susceptibility was evaluated by [...] Read more.
The aim of the present work was to genetically characterise cefotaxime-resistant enterobacteria isolated from community carriers in Bulgaria. In total, 717 faecal samples from children and adults in five medical centres in Sofia, Pleven and Burgas were examined. Antimicrobial susceptibility was evaluated by the disk diffusion method. blaESBL or plasmidic AmpC (pAmpC) genes were detected by PCR and sequencing. MLST and ERIC-PCR were used to detect clonal relatedness. Among the faecal samples, 140 cefotaxime-resistant enterobacteria were found. The most frequently detected species was Escherichia coli (77.9%, 109/140 samples), followed by Klebsiella pneumoniae (7.9%, 11/140). Among the isolates, blaCTX-M-15 (37.1%) was predominant, followed by blaCTX-M-3 (19.2%), blaCTX-M-14 (10%), and blaCTX-M-27 (4.3 %). Genes encoding pAmpC were observed in 11.4% (blaDHA-1, 16/140) and in 1.4% (blaCMY-2, 2/140). The frequency of ESBL and pAmpC producers among the subjects was 14.6% and 2.5%, respectively. No carbapenem-resistant isolates were found. Four main clonal complexes (CC131, CC10, CC38, and CC155) were detected among E. coli isolates. The most common type was ST131, phylogroup B2 (16.5%). The increased frequency of ESBL- and pAmpC-producing enterobacteria in the community is a prerequisite for treatment failures of the associated infections and a good background for further studies. Full article
(This article belongs to the Special Issue Mechanisms of Antimicrobial Resistance in Bacterial Pathogens)
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11 pages, 779 KB  
Article
Clinical Outcomes of Patients with AmpC-Beta-Lactamase-Producing Enterobacterales Bacteremia Treated with Carbapenems versus Non-Carbapenem Regimens: A Single-Center Study
by Orjowan Shalabi, Livnat Kashat, Omer Murik, Shoshana Zevin, Marc V. Assous and Eli Ben-Chetrit
Antibiotics 2024, 13(8), 709; https://doi.org/10.3390/antibiotics13080709 - 29 Jul 2024
Cited by 5 | Viewed by 3512
Abstract
Introduction: Bloodstream infections caused by AmpC-producing Enterobacterales pose treatment challenges due to the risk of AmpC overproduction and treatment failure. Current guidelines recommend carbapenems or cefepime as optimal therapy. We aimed to evaluate empiric and definitive non-carbapenem regimens for these infections. Methods: In [...] Read more.
Introduction: Bloodstream infections caused by AmpC-producing Enterobacterales pose treatment challenges due to the risk of AmpC overproduction and treatment failure. Current guidelines recommend carbapenems or cefepime as optimal therapy. We aimed to evaluate empiric and definitive non-carbapenem regimens for these infections. Methods: In a retrospective study from June 2014 to March 2023, adult bacteremic patients with Enterobacter cloacae complex strains and Morganella morganii were evaluated. Demographic, clinical and lab data and outcomes were assessed. Results: The cohort comprised 120 bacteremic patients, 17 receiving empiric carbapenem and 103 non-carbapenem regimens. Both groups had similar Charlson and Norton scores and previous antimicrobial exposure. The most common sources of bacteremia were urinary, abdominal and central-line-associated sources. Empiric non-carbapenem regimens (primarily piperacillin–tazobactam and cephalosporins) were not associated with recurrent bacteremia or 30-day mortality. Definitive regimens included mainly carbapenems (n = 41) and ciprofloxacin (n = 46). Beta-lactams were administered to 25 patients. Recurrent bacteremia and 30-day mortality rates were similar among treatment groups. Ciprofloxacin showed comparable outcomes to carbapenems, however, severity of illness among these patients was lower. Conclusions: Empiric and definitive non-carbapenem regimens for bacteremia with AmpC-producing organisms were not associated with treatment failure or increased 30-day mortality. Ciprofloxacin appears promising for selected, stable patients, potentially enabling early discharge. Full article
(This article belongs to the Special Issue Epidemiology and Mechanism of Bacterial Resistance to Antibiotics)
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