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15 pages, 4666 KB  
Article
Alleviation of Saline–Alkaline Stress in Alfalfa by a Consortium of Plant-Growth-Promoting Rhizobacteria
by Lingjuan Han, Yixuan Li, Zheng Ma, Bin Li, Yinping Liang, Peng Gao and Xiang Zhao
Plants 2025, 14(17), 2744; https://doi.org/10.3390/plants14172744 - 2 Sep 2025
Abstract
Soil salinization critically threatens global agricultural productivity by impairing plant growth and soil fertility. This study investigated the potential of a consortium, comprising Acinetobacter calcoaceticus DP25, Staphylococcus epidermidis DP28, and Enterobacter hormaechei DP29, to enhance the saline–alkali tolerance of alfalfa and improve soil [...] Read more.
Soil salinization critically threatens global agricultural productivity by impairing plant growth and soil fertility. This study investigated the potential of a consortium, comprising Acinetobacter calcoaceticus DP25, Staphylococcus epidermidis DP28, and Enterobacter hormaechei DP29, to enhance the saline–alkali tolerance of alfalfa and improve soil properties. The experiments comprised five germination treatments (saline control, each strain alone, consortium) and three pot treatments (non-saline control, saline control, consortium). Under saline–alkali stress, co-inoculation with the consortium significantly (p < 0.05) increased alfalfa seed germination rates, emergence rates, and biomass (shoot and root dry weight), while promoting root development. Physiological analyses revealed that the bacterial consortium mitigated stress-induced damage by enhancing photosynthetic efficiency, chlorophyll content, and antioxidant enzyme activities (superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT)), while decreasing malondialdehyde (MDA) levels. Moreover, the inoculant improved osmoprotectant accumulation (soluble sugars, soluble proteins, and proline) and modulated soil properties by reducing pH and electrical conductivity (EC), while elevating nutrient availability and soil enzyme activities. Correlation and principal component analyses (PCA) confirmed strong associations among improved plant growth, physiological traits, and soil health. These findings demonstrate that the bacterial consortium effectively alleviates saline–alkali stress in alfalfa by improving soil health, offering a sustainable strategy for ecological restoration and improving agricultural productivity in saline–alkali regions. Full article
(This article belongs to the Special Issue Horticultural Plant Physiology and Molecular Biology—2nd Edition)
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13 pages, 2986 KB  
Article
Endophyte Diversity and Resistance to Pine Wilt Disease in Coniferous Trees
by Shuting Zhao, Chao Wang, Qunqun Guo, Yanxin Pan, Meng Zhang, Huiyu Wang, Jiayi Yu, Ronggui Li and Guicai Du
Forests 2025, 16(9), 1403; https://doi.org/10.3390/f16091403 - 2 Sep 2025
Abstract
Pine wilt disease (PWD) is a serious forest disease caused by pine wood nematode (PWN). To examine the relationship between coniferous endophytes and PWD resistance, this study investigated endophytic bacterial and fungal communities in five conifer species: two Japanese black pine populations ( [...] Read more.
Pine wilt disease (PWD) is a serious forest disease caused by pine wood nematode (PWN). To examine the relationship between coniferous endophytes and PWD resistance, this study investigated endophytic bacterial and fungal communities in five conifer species: two Japanese black pine populations (Pinus thunbergii from Qingdao University, PQ, and Fushan Forest Park, PF), Chinese arborvitae (Platycladus orientalis, PO), cedar (Cedrus deodara, CD), and Masson pine (Pinus massoniana, PM). Results showed a strong correlation between endophytic microbial diversity and PWD resistance. PO with high PWD resistance hosted the most unique bacterial species, while PM with low PWD resistance had the fewest unique bacteria and significantly lower ACE and Shannon indices. At the bacterial genus level, dominant genera in resistant conifers often showed high nematocidal activity, whereas those in susceptible plants boosted nematode reproduction. PQ featured the unique dominant genus Pantoea, and PO’s unique Acinetobacter and the shared genus Bacillus (with CD) both displayed high toxicity to PWNs. In contrast, PF’s Pseudomonas and PM’s Stenotrophomonas significantly promoted nematode reproduction. Fungal community analysis revealed that the unique endophytic fungi in PQ are more abundant than those in PF, and the Shannon index of its endophytic fungi is comparable to that of CD and significantly higher than that of PF. PF’s dominant fungal genus Pestalotiopsis might facilitate nematode invasion, and its fungal Shannon index is significantly lower than PQ’s. Eight bacterial strains were isolated from these five conifer plants, with six highly nematocidal strains originating from PQ, CD, and PO. This study offers evidence that endophytic microbial communities critically influence PWD resistance, offering a microbial basis for developing resistant conifer cultivars through microbiome engineering. Full article
(This article belongs to the Section Forest Biodiversity)
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24 pages, 2071 KB  
Article
Increased Antimicrobial Consumption, Isolation Rate, and Resistance Profiles of Multi-Drug Resistant Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii During the COVID-19 Pandemic in a Tertiary Healthcare Institution
by Predrag Savic, Ljiljana Gojkovic Bukarica, Predrag Stevanovic, Teodora Vitorovic, Zoran Bukumiric, Olivera Vucicevic, Nenad Milanov, Vladimir Zivanovic, Ana Bukarica and Milos Gostimirovic
Antibiotics 2025, 14(9), 871; https://doi.org/10.3390/antibiotics14090871 - 29 Aug 2025
Viewed by 261
Abstract
Background: The aims of this paper are to examine the impact of the COVID-19 pandemic on the non-rational use of antibiotics and potential alterations in the antibiotic resistance profiles of multi-drug resistant (MDR) isolates of Klebsiella pneumoniae (KPN), Pseudomonas aeruginosa (PAE), and Acinetobacter [...] Read more.
Background: The aims of this paper are to examine the impact of the COVID-19 pandemic on the non-rational use of antibiotics and potential alterations in the antibiotic resistance profiles of multi-drug resistant (MDR) isolates of Klebsiella pneumoniae (KPN), Pseudomonas aeruginosa (PAE), and Acinetobacter baumannii (ABA). Material and Methods: This study was conducted at the tertiary University Hospital “Dr Dragisa Misovic-Dedinje” (Belgrade, Serbia) and was divided into three periods: pre-pandemic (1.4.2019–31.3.2020, period I), COVID-19 pandemic (1.4.2020–31.3.2021, period II), and COVID-19 pandemic-second phase (1.4.2021–31.3.2022, period III). Cultures were taken from each patient with clinically suspected infection (symptoms, biochemical markers of infection). All departments of the hospital were included in this study. Based on the source, all microbiological specimens were divided into 1° blood, 2° respiratory tract (tracheal aspirate, bronchoalveolar lavage fluid, throat, sputum), 3° central-line catheter, 4° urine, 5° urinary catheter, 6° skin and soft tissue, and 6° other (peritoneal fluid, drainage sample, bioptate, bile, incisions, fistulas, and abscesses). After the isolation of bacterial strains from the samples, an antibiotic sensitivity test was performed for each individual isolate with the automated Vitek® 2 COMPACT. Antibiotic consumption testing was performed by the WHO guideline equations (ATC/DDD). Results: A total of 2196 strains of KPN, PAE, and ABA from 41,144 hospitalized patients were isolated (23.6% of the number of total isolates). The number of ABA isolates statistically increased (p = 0.021), while the number of PAE isolates statistically decreased (p = 0.003) during the pandemic. An increase in the percentage of MDR strains was observed for KPN (p = 0.028) and PAE (p = 0.027). There has been an increase in the antibiotic resistance of KPN for piperacillin–tazobactam, the third and fourth generations of cephalosporins (ceftriaxone, ceftazidime, and cefepime), all carbapanems (imipenem, meropenem, and ertapenem), and levofloxacin; of PAE for imipenem; and of ABA for amikacin. Total antibiotic consumption increased (from 755 DBD to 1300 DBD, +72%), especially in the watch and reserve group of antibiotics. The highest increases were noted for vancomycin, levofloxacin, azithromycin, and meropenem. MV positively correlated with the increased occurrence of MDR KPN (r = 0.35, p = 0.009) and MDR PAE (r = 0.43, p = 0.009) but not for MDR ABA (r = 0.09, p = 0.614). There has been a statistically significant increase in the Candida sp. isolates, but the prevalence of Clostridium difficile infection remained unchanged. Conclusions: The COVID-19 pandemic has influenced the increase in total and MDR strains of KPN, ABA, and PAE and worsened their antibiotic resistance profiles. An increase in the consumption of both total and specific antibiotics was observed, mostly of fluoroquinolones and carbapenems. A positive correlation between the number of patients on MV and an increase in MDR KPN and MDR PAE strains was noted. It is necessary to adopt and demand the implementation of appropriate antimicrobial stewardship interventions to decrease the resistance of intrahospital pathogens to antibiotics. Full article
(This article belongs to the Special Issue Antimicrobial Stewardship in the Management of Bloodstream Infections)
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16 pages, 1905 KB  
Article
Microbiological and Clinical Characteristics of Pediatric Sepsis Patients with and without Septic Shock: A Retrospective Study at a Tertiary Pediatric Hospital in China
by Kai-Cheng Peng, Qin-Yuan Li, Lin Chen, Yan Zhao, Hui Liu, Zhen-Xuan Kong and Zheng-Xiu Luo
Children 2025, 12(9), 1146; https://doi.org/10.3390/children12091146 - 28 Aug 2025
Viewed by 165
Abstract
Background: Pediatric sepsis, a life-threatening condition, often progresses to septic shock. However, microbiological and clinical profiles between pediatric sepsis patients with and without septic shock remain underexplored. Methods: This retrospective cohort study included 1200 pediatric sepsis patients (Phoenix Sepsis Score ≥ 2) from [...] Read more.
Background: Pediatric sepsis, a life-threatening condition, often progresses to septic shock. However, microbiological and clinical profiles between pediatric sepsis patients with and without septic shock remain underexplored. Methods: This retrospective cohort study included 1200 pediatric sepsis patients (Phoenix Sepsis Score ≥ 2) from the Children’s Hospital of Chongqing Medical University between June 2018 and June 2023. Pediatric sepsis patients with septic shock were diagnosed based on the Phoenix Cardiovascular Score being ≥1. Clinical data and pathogens were taken from the electronic medical records and analyzed. Univariate and multivariable logistic regressions were conducted to identify the risk factors for septic shock. Results: Septic shock patients had longer hospital stays (14.8 vs. 12.0 days, p = 0.003) and higher mortality (17.4% vs. 4.7%, p < 0.001) when compared to non-septic-shock patients. Among these two groups, the pathogen prevalence revealed that bacterial pathogens dominated (48.9%), followed by viruses (10.3%). Acinetobacter baumannii, Escherichia coli, and Staphylococcus aureus remained the predominant pathogens; Pseudomonas aeruginosa and Mycoplasma pneumoniae were also increased. Combination antibiotic therapy was most frequent in patients with viral and fungal sepsis (79% and 86.5%, respectively). Patients with fungal sepsis had significantly longer hospital stays than those with viral sepsis (20.74 vs. 12.97 days, p = 0.017). Multivariable analysis identified that elevated lactate (OR = 1.49, 95% CI = 1.29–1.75) and pulmonary infection (OR = 2.25, 95% CI = 1.35–3.73) were independent risk factors for septic shock. Conclusions: Children with septic shock had higher mortality and prolonged hospitalization, with distinct microbiological profiles when compared with patients in the non-septic-shock group. Elevated lactate and presence of pulmonary infection are independent risk factors for septic shock. Early recognition of high-risk patients and tailored antimicrobial strategies are critical for improving outcomes. Full article
(This article belongs to the Section Pediatric Allergy and Immunology)
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13 pages, 810 KB  
Article
Optimization of 16S RNA Sequencing and Evaluation of Metagenomic Analysis with Kraken 2 and KrakenUniq
by Nasserdine Papa Mze, Cécile Fernand-Laurent, Sonnentrucker Maxence, Olfa Zanzouri, Solen Daugabel and Stéphanie Marque Juillet
Diagnostics 2025, 15(17), 2175; https://doi.org/10.3390/diagnostics15172175 - 27 Aug 2025
Viewed by 758
Abstract
Background/Objectives: 16S ribosomal RNA sequencing has, for several years, been the main means of identifying bacterial and archaeal species. Low-throughput Sanger sequencing is often used for the detection and identification of microbial species, but this technique has several limitations. The use of [...] Read more.
Background/Objectives: 16S ribosomal RNA sequencing has, for several years, been the main means of identifying bacterial and archaeal species. Low-throughput Sanger sequencing is often used for the detection and identification of microbial species, but this technique has several limitations. The use of high-throughput sequencers may be a good alternative to improve patient identification, especially for polyclonal infections and management. Kraken 2 and KrakenUniq are free, high-throughput tools providing a very rapid and accurate classification for metagenomic analyses. However, Kraken 2 can present false-positive results relative to KrakenUniq, which can be limiting in hospital settings requiring high levels of accuracy. The aim of this study was to establish an alternative next-generation sequencing technique to replace Sanger sequencing and to confirm that KrakenUniq is an excellent analysis tool that does not present false results relative to Kraken 2. Methods: DNA was extracted from reference bacterial samples for Laboratory Quality Controls (QCMDs) and the V2-V3 and V3-V4 regions of the 16S ribosomal gene were amplified. Amplified products were sequenced with the Illumina 16S Metagenomic Sequencing protocol with minor modifications to adapt and sequence an Illumina 16S library with a small 500-cycle nano-flow cell. The raw files (Fastq) were analyzed on a commercial Smartgene platform for comparison with Kraken 2 and KrakenUniq results. KrakenUniq was used with a standard bacterial database and with the 16S-specific Silva138, RDP11.5, and Greengenes 13.5 databases. Results: Seven of the eight (87.5%) QCMDs were correctly sequenced and identified by Sanger sequencing. The remaining QCMD, QCMD6, could not be identified through Sanger sequencing. All QCMDs were correctly sequenced and identified by MiSeq with the commercial Smartgene analysis platform. QCMD6 contained two bacteria, Acinetobacter and Klebsiella. KrakenUniq identification results were identical to those of Smartgene, whereas Kraken 2 yielded 25% false-positive results. Conclusions: If Sanger identification fails, MiSeq with a small nano-flow cell is a very good alternative for the identification of bacterial species. KrakenUniq is a free, fast, and easy-to-use tool for identifying and classifying bacterial infections. Full article
(This article belongs to the Section Pathology and Molecular Diagnostics)
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23 pages, 3322 KB  
Article
Genetic Diversity, Extended-Spectrum Beta-Lactamase (ESBL) Screening, and Potential Public Health Implications of Gram-Negative Bacteria Recovered from Man-Made Lakes and Surrounding Vegetables
by Ahou Cinthia Inès Yebouet, Kouakou Romain Fossou, Zaka Ghislaine Claude Kouadjo-Zézé, Anthony Ifeanyi Okoh and Adolphe Zézé
Microorganisms 2025, 13(9), 1997; https://doi.org/10.3390/microorganisms13091997 - 27 Aug 2025
Viewed by 310
Abstract
The emergence and dissemination of extended-spectrum beta-lactamase (ESBL)-producing bacteria pose a major public health threat, necessitating a One Health approach to addressing this threat. Thus, the diversity, ESBL production, and potential public health implications of Gram-negative bacteria recovered from man-made lakes and surrounding [...] Read more.
The emergence and dissemination of extended-spectrum beta-lactamase (ESBL)-producing bacteria pose a major public health threat, necessitating a One Health approach to addressing this threat. Thus, the diversity, ESBL production, and potential public health implications of Gram-negative bacteria recovered from man-made lakes and surrounding lettuce in Yamoussoukro, Côte d’Ivoire were assessed in this study. Also, the lakes’ physicochemical parameters were assessed and correlated with bacteria community using Pearson correlation. A total of 68 Gram-negative bacterial isolates were recovered from the samples and identified via 16S rDNA gene sequencing. Phylogenetic analysis suggested multiple genus-/species-level variations within the isolates. Escherichia coli was the most prevalent in lake water (39.5%), while Acinetobacter was the dominant genus in lettuce (30%). E. coli isolates showed high resistance to ampicillin (90.9%), cefepime (72.7%), cefotaxime (68.2%), and aztreonam (63.6%). Moreover, ESBL production was confirmed in E. coli isolates (22.05%), predominantly mediated by the blaCTX-M gene. Multidrug-resistant phenotypes were widespread, yielding similar multiple antibiotic resistance index (MARI) values in water (0.27–0.63) and lettuce (0.27–0.81). These data indicate high environmental contamination, which unfortunately is not being taken into account by lettuce producers according to an interview. Statistical analyses showed a significant relationship between bacterial diversity and lakes’ physicochemical parameters, including dissolved oxygen, pH, and turbidity. The basic education level of farmers, the prevalence of ESBL-producing E. coli, and the high prevalence of MDR Gram-negative bacteria in both environmental and crop sources in Yamoussoukro underscore the need for both integrated surveillance and management strategies to mitigate potential microbial public health risks within a One Health framework. Full article
(This article belongs to the Special Issue Bacterial Antibiotic Resistance, Second Edition)
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13 pages, 538 KB  
Article
The Skin Microbiome Profile of Contact Sports Athletes—Focus on Sexual Dimorphism and Athlete–Non-Athlete Differences
by Irina Kalabiska, Dorina Annar, Gergely Babszky, Matyas Jokai, Zoltan Borbas, Gergely Hajdu, Fanny Zselyke Ratz-Sulyok, Csilla Jang-Kapuy, Gergely Palinkas, Harjit Pal Bhattoa and Annamaria Zsakai
Sports 2025, 13(9), 288; https://doi.org/10.3390/sports13090288 - 26 Aug 2025
Viewed by 976
Abstract
Background: Athletes’ skin is exposed to increased microbial challenges due to rigorous physical activity, perspiration, constant “skin-to-skin” contact, frequent showering, use of hygiene products, and environmental factors present in training settings. This study aims to characterize the skin microbiome communities of young wrestlers [...] Read more.
Background: Athletes’ skin is exposed to increased microbial challenges due to rigorous physical activity, perspiration, constant “skin-to-skin” contact, frequent showering, use of hygiene products, and environmental factors present in training settings. This study aims to characterize the skin microbiome communities of young wrestlers and kickboxers in comparison with their non-athlete age-peers. Methods: A total of 56 combat sport athletes (30 males and 26 females, mean age ± SD = 18.2 ± 1.5 years) and 25 non-athlete youths (control group: 13 males and 12 females, mean age ± SD = 19.8 ± 1.2 years) voluntarily consented to participate in the study conducted by our research team in 2023 and 2024. The skin microbiome analysis involved standardized sampling, DNA isolation, molecular sequencing, and bioinformatic analysis, thus enabling detailed characterization and comparison of the skin microbial community in contact sports athletes and the control group. Results: Our results revealed notable sexual dimorphism in the skin microbiome composition of youth. Males showed a higher relative abundance of bacterial genera associated with nosocomial infections and respiratory diseases, while females had more skin inflammation- and infection-related genera (relative abundances in males vs. in females: Corynebacterium—12.0 vs. 7.2; Luteimonas—4.4. vs. 1.4; Paracoccus—8.8 vs. 5.0; Psychrobacter—6.3 vs. 4.4; Cutibacterium—6.4 vs. 11.4; Kocuria—1.6 vs. 3.9; Micrococcus—5.8 vs. 8.5; Pseudomonas—1.2 vs. 3.4; Streptococcus 3.3 vs. 6.2). We also found skin microbiome differences between athletes and non-athletes in both sexes: wrestlers, who experience frequent skin-to-skin contact and wear less covering sportswear, had microbiome profiles distinct from both kickboxers and non-athletes (relative abundances in athletes vs. in non-athletes: Psychrobacter—7.3 vs. 0.4; Staphylococcus 9.5 vs. 18.5; predominance of genera by sports type: relative abundance of Cutibacterium and Streptococcus was higher in kickboxers, and relative abundance of Acinetobacter, Enhydrobacter, Micrococcus, and Enhydrobacter was higher in wrestlers). Bacteria linked to skin infections (e.g., Aliterella, Arthrobacter, and Empedobacter) were present in around 30% of wrestlers and kickboxers but were absent in the control group. Conclusions: These results underscore the heightened risk of skin infections in contact sports and highlight the importance of regular microbiome monitoring and hygiene protocols among young athletes. Full article
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22 pages, 4867 KB  
Article
Comparative Analysis of Compound Probiotics, Seasonal Variation, and Age on Gut Microbial Composition and Function in Endangered Forest Musk Deer
by Feng Jiang, Pengfei Song, Zhenyuan Cai, Guosheng Wu, Shunfu He, Haifeng Gu, Hongmei Gao and Tongzuo Zhang
Microorganisms 2025, 13(9), 1991; https://doi.org/10.3390/microorganisms13091991 - 26 Aug 2025
Viewed by 369
Abstract
Due to persistent poaching and habitat fragmentation, wild forest musk deer (Moschus berezovskii) in China have sharply declined; although captive breeding helps, frequent gut diseases limit further expansion. This study used high-throughput 16S rRNA sequencing to analyze the effects of age, [...] Read more.
Due to persistent poaching and habitat fragmentation, wild forest musk deer (Moschus berezovskii) in China have sharply declined; although captive breeding helps, frequent gut diseases limit further expansion. This study used high-throughput 16S rRNA sequencing to analyze the effects of age, season variation, and compound probiotics on the gut microbiota of captive individuals. The results demonstrated that compound probiotics exerted a significantly greater influence on gut microbial composition, α-diversity, and functional variation compared to the effects of age or seasonal factors. β-diversity analysis confirmed greater differences between probiotic-treated and control groups than among age or seasonal groups. Microbial community assembly was mainly driven by deterministic processes, with stochastic processes also playing a role in winter. Compound probiotics markedly reshaped dominant bacterial taxa at both phylum and genus levels, with Acinetobacter identified as a key biomarker. They also significantly modulated metabolic and phenotypic traits, decreasing functions related to Gram-positive and aerobic bacteria while enhancing those linked to Gram-negative characteristics. Environmental correlation analysis further demonstrated that compound probiotics exerted a stronger influence than both age and seasonal factors. The findings underscore the value of dietary and probiotic strategies for enhancing gut health and resilience in endangered forest musk deer. Full article
(This article belongs to the Section Gut Microbiota)
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32 pages, 1122 KB  
Review
Carbapenem-Resistant Acinetobacter baumannii: Virulence Factors, Molecular Epidemiology, and Latest Updates in Treatment Options
by Theodoros Karampatakis, Katerina Tsergouli and Payam Behzadi
Microorganisms 2025, 13(9), 1983; https://doi.org/10.3390/microorganisms13091983 - 26 Aug 2025
Viewed by 739
Abstract
Acinetobacter baumannii is a Gram-negative, non-motile pathogen commonly associated with healthcare settings. It is capable of causing severe infections, particularly in immunocompromised and critically ill individuals, and is linked to poor clinical outcomes. Infections caused by carbapenem-resistant A. baumannii (CRAB) represent a major [...] Read more.
Acinetobacter baumannii is a Gram-negative, non-motile pathogen commonly associated with healthcare settings. It is capable of causing severe infections, particularly in immunocompromised and critically ill individuals, and is linked to poor clinical outcomes. Infections caused by carbapenem-resistant A. baumannii (CRAB) represent a major public health concern due to limited treatment options and high resistance rates. Several virulence determinants contribute to CRAB’s pathogenicity, including capsular exopolysaccharide (CPS), lipopolysaccharide (LPS), lipooligosaccharide (LOS), efflux pumps, outer membrane proteins (OMPs), pili, metal acquisition systems, two-component regulatory systems (TCSs), and secretion systems (SSs). The dominant resistance mechanism in CRAB involves the production of carbapenemases, most notably oxacillinase-23 (OXA-23) and metallo-β-lactamases (MBLs) such as Verona integron-encoded MBL (VIM) and New Delhi MBL (NDM). Accurate identification of these resistance mechanisms is crucial for guiding effective antimicrobial therapy. Potential treatment options include older agents like polymyxins, ampicillin–sulbactam, high-dose carbapenems, tigecycline, and minocycline, along with newer antimicrobials such as eravacycline, cefiderocol, and aztreonam–avibactam. This review aims to explore the virulence mechanisms and molecular pathogenesis of CRAB, while also presenting recent developments in its epidemiology and available antimicrobial therapies. Full article
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24 pages, 1750 KB  
Systematic Review
A 15-Year One Health Approach to Antimicrobial Resistance in Kuwait from Hospitals to Environmental Contexts: A Systematic Review
by Ahmad Al-Dhumair, Mohammad Al-Hasan, Hanan Al-Khalaifah and Qadriya Al-Mutawa
Life 2025, 15(9), 1344; https://doi.org/10.3390/life15091344 - 25 Aug 2025
Viewed by 462
Abstract
Kuwait has reported a problematic increase in the prevalence of Antimicrobial Resistance (AMR). However, the absence of studies that analyze AMR from combined human, agricultural (animal), and environmental domains limits our ability to assess the extent of the problem in Kuwait. Therefore, this [...] Read more.
Kuwait has reported a problematic increase in the prevalence of Antimicrobial Resistance (AMR). However, the absence of studies that analyze AMR from combined human, agricultural (animal), and environmental domains limits our ability to assess the extent of the problem in Kuwait. Therefore, this systematic review provides a comprehensive insight into the AMR status in Kuwait regarding humans, agriculture (animals), and the environment from the perspective of the One Health approach. A systematic search was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines to identify the relevant literature on AMR in Kuwait. Multiple online electronic databases, including the Cochrane Library, Google Scholar, Web of Science, PubMed, BioMed Central (BMC), and Scopus, were searched to perform a narrative synthesis and meta-analysis. Twenty-eight studies published between 2009 and 2024 were included in this study. Domain-wise distribution varied, with 11 studies related to clinical settings, 11 to the environment, 4 to agricultural (animal), and 2 to both clinical and community settings. The narrative synthesis indicated a high occurrence of AMR bacteria in human, agriculture (animal), and environmental domains. In human domains, the dominant AMR isolates belonged to four Gram-negative species: E. coli, K. pneumoniae, P. aeruginosa, and Acinetobacter baumannii. In agriculture (animals), Salmonella isolates from poultry display high resistance to cefotaxime, ampicillin, and amoxicillin. Camel milk analyses revealed that 80% of bacterial isolates are resistant to antibiotics such as penicillin, tetracyclines, and carbapenems. An environmental analysis of sewage, seawater, sediment, and aerosol samples documented widespread antibiotic resistance genes (ARGs) with resistance mechanisms such as extended-spectrum beta-lactamase, carbapenemases, and colistin. The cross-domain analysis identified the overlapping of ARGs. Regarding the One Health approach, none of the studies used this approach to interlink these sectors. Meanwhile, the meta-analysis indicated a high resistance rate in humans (34.05%, 95% CI (22.81 to 46.27, p < 0.0001, I2 = 98.94%)), agriculture (animals) (67.42%, 95% CI (30.30 to 94.93, p < 0.0001, I2 = 97.40%)), and environment (69.86%, 95% CI (48.80 to 87.26, p < 0.0001, I2 = 98.78%)). The reported spread of AMR and the overlap of resistance genes among isolates across the domains demonstrate the interconnected nature of AMR in Kuwait. These findings underscore the need to adopt the One Health approach to strengthen surveillance, implement control measures, and enhance public education strategies to address the complexity of AMR challenges in Kuwait. Full article
(This article belongs to the Section Pharmaceutical Science)
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12 pages, 3430 KB  
Article
Deciphering Bacterial Community Succession and Pathogen Dynamics in ICU Ventilator Circuits Through Full-Length 16S rRNA Sequencing for Mitigating the Risk of Nosocomial Infections
by Hsin-Chi Tsai, Jung-Sheng Chen, Gwo-Jong Hsu, Bashir Hussain, I-Ching Lin, Tsui-Kang Hsu, Jing Han, Shih-Wei Huang, Chin-Chia Wu and Bing-Mu Hsu
Microorganisms 2025, 13(9), 1982; https://doi.org/10.3390/microorganisms13091982 - 25 Aug 2025
Viewed by 365
Abstract
The rapid evolution of ventilators and their circuits, coupled with varying maximum usage durations set by different hospitals globally, poses a significant risk for the proliferation and transmission of nosocomial infections in intensive care settings. This study investigated temporal changes in bacterial community [...] Read more.
The rapid evolution of ventilators and their circuits, coupled with varying maximum usage durations set by different hospitals globally, poses a significant risk for the proliferation and transmission of nosocomial infections in intensive care settings. This study investigated temporal changes in bacterial community structure and predicted metabolic functions in ventilator circuits over a three-week period, with a specific focus on ESKAPE pathogens. The results of full-length 16S rRNA sequencing revealed dynamic shifts in bacterial communities, with an increased bacterial diversity and unique species prevalence in week-2 compared to week-1 and week-3. However, a marked emergence of pathogenic bacteria, including Serratia marcescens and Chryseobacterium indologenes, was observed in week-3 compared to week-1 and week-2. Additionally, the abundance of ESKAPE pathogens, including Klebsiella pneumoniae and Acinetobacter baumannii, was higher in week-3 compared to week-1 and week-2. Furthermore, the PCR analysis revealed a higher detection rate of Pseudomonas aeruginosa and K. pneumoniae in week-3 than in the previous weeks. FAPROTAX analysis further revealed a high abundance of specific functions associated with the pathogens of pneumonia, nosocomial, and septicemia in week-3 compared to the other two weeks, suggesting a shift toward more virulent or opportunistic pathogens with increased utilization of ventilator circuits. These findings highlight the microbial risks associated with prolonged use of ventilator circuits, underscoring the need for continuous microbial surveillance throughout their usage, and provide a foundation for optimizing infection control strategies in intensive care settings. Full article
(This article belongs to the Special Issue The Molecular Epidemiology of Infectious Diseases)
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35 pages, 4318 KB  
Article
Episode- and Hospital-Level Modeling of Pan-Resistant Healthcare-Associated Infections (2020–2024) Using TabTransformer and Attention-Based LSTM Forecasting
by Nicoleta Luchian, Camer Salim, Alina Plesea Condratovici, Constantin Marcu, Călin Gheorghe Buzea, Mădalina Nicoleta Matei, Ciprian Adrian Dinu, Mădălina Duceac (Covrig), Eva Maria Elkan, Dragoș Ioan Rusu, Lăcrămioara Ochiuz and Letiția Doina Duceac
Diagnostics 2025, 15(17), 2138; https://doi.org/10.3390/diagnostics15172138 - 25 Aug 2025
Viewed by 375
Abstract
Background: Pan-drug-resistant (PDR) Acinetobacterinfections are an escalating ICU threat, demanding both patient-level triage and facility-wide forecasting. Objective: The aim of this study was to build a dual-scale AI framework that (i) predicts PDR status at infection onset and (ii) forecasts hospital-level [...] Read more.
Background: Pan-drug-resistant (PDR) Acinetobacterinfections are an escalating ICU threat, demanding both patient-level triage and facility-wide forecasting. Objective: The aim of this study was to build a dual-scale AI framework that (i) predicts PDR status at infection onset and (ii) forecasts hospital-level PDR burden through 2027. Methods: We retrospectively analyzed 270 Acinetobacter infection episodes (2020–2024) with 65 predictors spanning demographics, timelines, infection type, resistance-class flags, and a 25-drug antibiogram. TabTransformer and XGBoost were trained on 2020–2023 episodes (n = 210), evaluated by stratified 5-fold CV, and externally tested on 2024 episodes (n = 60). Metrics included AUROC, AUPRC, accuracy, and recall at 90% specificity; AUROC was optimism-corrected via 0.632 + bootstrap and DeLong-tested for drift. SHAP values quantified feature impact. Weekly PDR incidence was forecast with an attention–LSTM model retrained monthly (200 weekly origins, 4-week horizon) and benchmarked against seasonal-naïve, Prophet, and SARIMA models (MAPE and RMSE). Quarterly projections (TFT-lite) extended forecasts to 2027. Results: The CV AUROC was 0.924 (optimism-corrected 0.874); an ensemble of TabTransformer + XGBoost reached 0.958. The 2024 AUROC fell to 0.586 (p < 0.001), coinciding with a PDR prevalence drop (75→38%) and three covariates with PSIs > 1.0. Isotonic recalibration improved the Brier score from 0.326 to 0.207 and yielded a net benefit equivalent to 26 unnecessary isolation-days averted per 100 ICU admissions at a 0.20 threshold. SHAP highlighted Ampicillin/Sulbactam resistance, unknown acquisition mode, and device-related infection as dominant drivers. The attention–LSTM achieved a median weekly MAE of 0.10 (IQR: 0.028–0.985) vs. 1.00 for the seasonal-naïve rule, outperforming it on 48.5% of weeks and surpassing Prophet and SARIMA (MAPE = 6.2%, RMSE = 0.032). TFT-lite projected a ≥ 25% PDR tipping point in 2025 Q1 with a sustained rise in 2027. Conclusions: The proposed framework delivers explainable patient-level PDR risk scores and competitive 4-week and multi-year incidence forecasts despite temporal drift, supporting antimicrobial stewardship and ICU capacity planning. Shrinkage and bootstrap correction were applied to address the small sample size (EPV = 2.1), which poses an overfitting risk. Continuous recalibration and multi-center validation remain priorities. Full article
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
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11 pages, 714 KB  
Article
Active Microbiological Surveillance for Contrasting Multi-Drug-Resistant Pathogens: Comparison Between a Multiplex Real-Time PCR Method and Culture
by Gaetano Maugeri, Maddalena Calvo, Guido Scalia and Stefania Stefani
Diagnostics 2025, 15(17), 2128; https://doi.org/10.3390/diagnostics15172128 - 22 Aug 2025
Viewed by 378
Abstract
Background/Objectives. Multi-drug-resistant (MDR) microorganisms pose a significant challenge in healthcare settings, particularly with beta-lactam-resistant Gram-negative bacteria and glycopeptide-resistant enterococci. Culture represents the most reliable technique in determining their presence within surveillance swabs. However, it requires a long time-to-result (TTR) and shows low [...] Read more.
Background/Objectives. Multi-drug-resistant (MDR) microorganisms pose a significant challenge in healthcare settings, particularly with beta-lactam-resistant Gram-negative bacteria and glycopeptide-resistant enterococci. Culture represents the most reliable technique in determining their presence within surveillance swabs. However, it requires a long time-to-result (TTR) and shows low sensitivity. Molecular techniques integrate diagnostic procedures, allowing TTR reduction and precise identification of genes. Methods. During our usual surveillance campaign, we had the opportunity to evaluate the Allplex Entero-DR assay (Seegene Inc., Seoul, Republic of Korea) and the Entero-DR Plus assay (Arrow Diagnostics srl, Genova, Italy) molecular kits for the detection of extended-β-lactamases (ESBL), carbapenem- and vancomycin-resistant genes, as well as Acinetobacter spp. and Pseudomonas aeruginosa spp. identification directly from rectal swabs. A comparison between these tests and the culture-based routine completed the study. Results. The analysis included 300 rectal swabs from the University Hospital Policlinico (Catania, Italy). One hundred and eighty-eight samples (62.6%) resulted as positive for at least one Allplex™ target, reaching optimal sensitivity and negative predictive value (100%). Our results underlined the ubiquitous blaCTX-M and van genes presence and demonstrated the diffusion of double-carbapenemases genes and metallo-β-lactamases-producing strains. In our epidemiological setting, few data were collected about carbapenem-resistant P. aeruginosa and Acinetobacter spp., which require further evaluations on simultaneous respiratory colonization and higher sample numbers. Conclusions. Our analysis highlighted the importance of combining conventional and advanced diagnostic methods in investigating MDR pathogens. The right approach should be based on the prevalence and variability of resistance mechanisms within a specific epidemiological area. Remarkably, molecular screenings may exclude negative samples within high-risk areas due to a significant negative predictive value. Full article
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19 pages, 1877 KB  
Article
Abscopal Brain Proteomic Changes Associated with Microbiome Alterations Induced by Gastrointestinal Acute Radiation Syndrome in Swine
by Kathleen Hatch, Timothy S. Horseman, Babita Parajuli, Erin K. Murphy, Robert N. Cole, Robert N. O’Meally, Daniel P. Perl, David M. Burmeister and Diego Iacono
Int. J. Mol. Sci. 2025, 26(17), 8121; https://doi.org/10.3390/ijms26178121 - 22 Aug 2025
Viewed by 212
Abstract
Emerging research highlights the gut microbiota’s critical role in modulating brain activity via the gut–brain axis. This study explores whether targeted gastrointestinal irradiation induces abscopal effects on the brain proteome, revealing microbiota-mediated neurobiological changes. Male Sinclair minipigs were randomized to receive either sham [...] Read more.
Emerging research highlights the gut microbiota’s critical role in modulating brain activity via the gut–brain axis. This study explores whether targeted gastrointestinal irradiation induces abscopal effects on the brain proteome, revealing microbiota-mediated neurobiological changes. Male Sinclair minipigs were randomized to receive either sham treatment (n = 6) or 8 Gy lower hemibody (gut-targeted) irradiation (n = 5). Over 14 days, rectal swabs were collected to monitor microbiota dynamics, followed by frontal cortex proteomic analysis. Irradiation altered gut microbiota composition, notably reducing Chlamydiae and Firmicutes phyla, while increasing Coriobacteriaceae and Acinetobacter. Proteomic analysis identified 75 differentially abundant proteins in the frontal cortex, including a significant decrease in pannexin-1 (PANX1), suggesting modulation of the NLRP3 inflammasome pathway. Functional enrichment analysis revealed immune and neurotransmission-related changes linked to microbial shifts. These results demonstrate that gut-targeted radiation can remotely affect brain protein expression, emphasizing the microbiota’s role in neuroimmune regulation and pointing to novel therapeutic opportunities in gut–brain axis disorders. Full article
(This article belongs to the Special Issue Emerging Roles of the Gut-Brain Axis (GBA) in Health and Disease)
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15 pages, 2355 KB  
Article
Biodegradation of Carbon Tetrachloride in Groundwater: Microbial Community Shifts and Functional Genes Involvement in Enhanced Reductive Dechlorination
by Zhengwei Liu, Mingbo Sun, Wei Wang, Shaolei Zhao, Yan Xie, Xiaoyu Lin, Jingru Liu and Shucai Zhang
Toxics 2025, 13(8), 704; https://doi.org/10.3390/toxics13080704 - 21 Aug 2025
Viewed by 277
Abstract
Carbon tetrachloride (CT) is a toxic volatile chlorinated hydrocarbon, posing a serious hazard to ecosystem and human health. This study discussed the bioremediation possibility of groundwater contaminated by CT. Enhanced reductive dechlorination bioremediation (ERD) was used to promote the reductive dechlorination process of [...] Read more.
Carbon tetrachloride (CT) is a toxic volatile chlorinated hydrocarbon, posing a serious hazard to ecosystem and human health. This study discussed the bioremediation possibility of groundwater contaminated by CT. Enhanced reductive dechlorination bioremediation (ERD) was used to promote the reductive dechlorination process of CT by adding yeast extract as a supplementary electron donor. The microcosm samples of the Control and Experi group were setup in the experiment, and the CT degradation efficiency and microbial community structure changes over 150 days were monitored. The results showed that the Experi group achieved complete degradation of CT within 40 days, while the control group had no significant change. By analyzing the physical and chemical indexes such as VFAs, sulfate ions, oxidation–reduction potential, pH value and so on, the key changes in the degradation process of CT were revealed. Microbial community analysis showed that specific microorganisms such as Acinetobacter johnsonii, Aeromonas media and Enterobacter mori played a significant role in the degradation of CT. They may produce hydrogen through fermentation to provide electron donors for the reductive dechlorination of CT. In addition, the genes of reductive dehalogenase synthase related to CT degradation were also identified, which provided molecular evidence for understanding the biodegradation mechanism of CT. The results deliver a scientific basis for optimizing the bioremediation strategy of CT-contaminated groundwater. Full article
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