Search

With the emergence of multidrug-resistant bacteria, the World Health Organization published a catalog of microorganisms urgently needing new antibiotics, with the carbapenem-resistant Acinetobacter baumannii designated as "critical". Such isolates, frequently detected in healthcare settings, pose a global pandemic threat. One way to facilitate a systemic view of bacterial metabolism and allow the development of new therapeutics is to apply constraint-based modeling. Here, we developed a versatile workflow to build high-quality and simulation-ready genome-scale metabolic models. We applied our workflow to create a metabolic model for A. baumannii and validated its predictive capabilities using experimental nutrient utilization and gene essentiality data. Our analysis showed that our model iACB23LX could recapitulate cellular metabolic phenotypes observed during in vitro experiments, while positive biomass production rates were observed and experimentally validated in various growth media. We further defined a minimal set of compounds that increase A. baumannii's cellular biomass and identified putative essential genes with no human counterparts, offering new candidates for future antimicrobial development. Finally, we assembled and curated the first collection of metabolic reconstructions for distinct A. baumannii strains and analyzed their growth characteristics. The presented models are in a standardized and well-curated format, enhancing their usability for multi-strain network reconstruction. Author summary: The emergence of multidrug-resistant bacteria, particularly carbapenem-resistant Acinetobacter baumannii, has become a severe global health threat. This pressing issue necessitated the development of new antibiotics, as highlighted by the World Health Organization. To address this need, we aimed to create comprehensive metabolic models to better understand bacterial metabolism and aid in developing novel therapeutic strategies. In this study, we developed a versatile workflow to construct high-quality, simulation-ready genome-scale metabolic models for bacterial pathogens. Applying this workflow, we constructed a metabolic model for A. baumannii and validated its accuracy using experimental data. The model successfully replicated observed metabolic phenotypes and identified essential genes without human counterparts, suggesting potential targets for new antibiotics. Additionally, we assembled and curated the first collection of metabolic reconstructions for distinct A. baumannii strains, analyzing their growth characteristics. These standardized and well-curated models enhance usability, facilitating multi-strain network reconstruction and further research. These findings provide a robust tool for understanding A. baumannii's metabolism, guiding the development of new antimicrobial therapies. [ABSTRACT FROM AUTHOR]

The quorum sensing (QS) system mediated by the abaI gene in Acinetobacter baumannii is crucial for various physiological and pathogenic processes. In this study, we constructed a stable markerless abaI knockout mutant (ΔabaI) strain using a pEXKm5-based allele replacement method to investigate the impact of abaI on A. baumannii. Proteomic analysis revealed significant alterations in protein expression between the wild type (WT) and ΔabaI mutant strains, particularly in proteins associated with membrane structure, antibiotic resistance, and virulence. Notably, the downregulation of key outer membrane proteins such as SurA, OmpA, OmpW, and BamA suggests potential vulnerabilities in outer membrane integrity, which correlate with structural abnormalities in the ΔabaI mutant strain, including irregular cell shapes and compromised membrane integrity, observed by scanning and transmission electron microscopy. Furthermore, diminished expression of regulatory proteins such as OmpR and GacA-GacS highlights the broader regulatory networks affected by abaI deletion. Functional assays revealed impaired biofilm formation and surface-associated motility in the mutant strain, indicative of altered colonization capabilities. Interestingly, the mutant showed a complex antibiotic susceptibility profile. While it demonstrated increased susceptibility to membrane-targeting antibiotics, its response to beta-lactams was more nuanced. Despite increased expression of metallo-beta-lactamase (MBL) superfamily proteins and DcaP-like protein, the mutant unexpectedly showed lower MICs for carbapenems (imipenem and meropenem) compared to the wild-type strain. This suggests that abaI deletion affects antibiotic susceptibility through multiple, potentially competing mechanisms. Further investigation is needed to fully elucidate the interplay between quorum sensing, antibiotic resistance genes, and overall antibiotic susceptibility in A. baumannii. Our findings underscore the multifaceted role of the abaI gene in modulating various cellular processes and highlight its significance in A. baumannii physiology, pathogenesis, and antibiotic resistance. Targeting the abaI QS system may offer novel therapeutic strategies for this clinically significant pathogen. [ABSTRACT FROM AUTHOR]

Background: A previous study highlighted the role of antibiotic-induced dysbiosis in the tick microbiota, facilitating the transstadial transmission of Babesia microti from nymph to adult in Haemaphysalis longicornis. This study builds on previous findings by analyzing sequence data from an earlier study to investigate bacterial interactions that could be linked to enhanced transstadial transmission of Babesia in ticks. The study employed antibiotic-treated (AT) and control-treated (CT) Haemaphysalis longicornis ticks to investigate shifts in microbial community assembly. Network analysis techniques were utilized to assess bacterial interactions, comparing network centrality measures between AT and CT groups, alongside studying network robustness and connectivity loss. Additionally, functional profiling was conducted to evaluate metabolic diversity in response to antibiotic treatment. Results: The analysis revealed notable changes in microbial community assembly in response to antibiotic treatment. Antibiotic-treated (AT) ticks displayed a greater number of connected nodes but fewer correlations compared to control-treated (CT) ticks, indicating a less interactive yet more connected microbial community. Network centrality measures such as degree, betweenness, closeness, and eigenvector centrality, differed significantly between AT and CT groups, suggesting alterations in local network dynamics due to antibiotic intervention. Coxiella and Acinetobacter exhibited disrupted connectivity and roles, with the former showing reduced interactions in AT group and the latter displaying a loss of connected nodes, emphasizing their crucial roles in microbial network stability. Robustness tests against node removal showed decreased stability in AT networks, particularly under directed attacks, confirming a susceptibility of the microbial community to disturbances. Functional profile analysis further indicated a higher diversity and richness in metabolic capabilities in the AT group, reflecting potential shifts in microbial metabolism as a consequence of antimicrobial treatment. Conclusions: Our findings support that bacterial interaction traits boosting the transstadial transmission of Babesia could be associated with reduced colonization resistance. The disrupted microbial interactions and decreased network robustness in AT ticks suggest critical vulnerabilities that could be targeted for managing tick-borne diseases. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a critical opportunistic pathogen associated with nosocomial infections. The high rates of antibiotic-resistance acquisition make most antibiotics ineffective. Thus, new medical countermeasures are urgently needed. Outer membrane proteins (OMPs) are prime candidates for developing novel drug targets and antibacterial strategies. However, there are substantial gaps in our knowledge of A. baumannii OMPs. This study reports the impact of OmpA-like protein on bacterial physiology and virulence in A. baumannii strain AB5075. We found that PsaB (ABUW_0505) negatively correlates to stress tolerance, while ArfA (ABUW_2730) significantly affects bacterial stiffness, cell shape, and cell envelope thickness. Furthermore, we expand our knowledge on YiaD (ABUW_3045), demonstrating structural and virulence roles of this porin, in addition to meropenem resistance. This study provides solid foundations for understanding how uncharacterized OMPs contribute to A. baumannii's physiological and pathological processes, aiding the development of innovative therapeutic strategies against A. baumannii infections.We provide foundations for understanding how the outer membrane porins PsaB, ArfA, and YiaD contribute to A. baumannii physiology and pathology, potentially aiding the development of novel therapeutic strategies. [ABSTRACT FROM AUTHOR]

Introduction and aim of study: Chronic wounds are increasingly challenging global healthcare. These wounds, which take over 3 months to heal, are complicated by untreated infections and the formation of biofilm, hindering healing and antibiotic effectiveness. To tackle these issues, new treatments like bacteriophage therapy are being explored. Bacteriophages, viruses that target bacteria, offer promise in overcoming antibiotic resistance. However, their use presents challenges that need to be addressed. Material and methods: Our review is based on the analysis of materials collected in Pubmed, Elsevier and other scientific articles using keywords: “chronic wound”, “chronic wounds infection”, “biofilms”, “MDR”, “bacteriophage”, “phage therapy”. Conclusions: The rise of chronic wounds due to resistant infections poses a significant challenge for patients and healthcare systems. Multidrug-resistant bacteria, often forming biofilm, evade current treatments, urging the search for alternatives. Phage therapy, showing efficacy against various stubborn infections, including those from surgery or diabetes, gains attention. Advanced delivery systems enhance targeted treatment, while phage cocktails improve effectiveness, especially against multiple resistant strains. Safety is generally observed, but larger trials are needed. Though not a replacement for antibiotics, phage therapy offers hope, needing robust clinical validation. While challenges exist, its societal, commercial, and economic benefits suggest a promising future beyond clinical use. [ABSTRACT FROM AUTHOR]

A growing increase in the number of serious infections caused by multidrug resistant bacteria (MDR) is challenging our society. Despite efforts to discover novel therapeutic options, few antibiotics targeting MDR have been approved by the Food and Drug Administration (FDA). Lactic acid bacteria have emerged as a promising therapeutic alternative due to their demonstrated ability to combat MDR pathogens in vitro. Our previous co-culture studies showed Lacticaseibacillus rhamnosus CRL 2244 as having a potent killing effect against carbapenem-resistant Acinetobacter baumannii (CRAB) strains. Here we report that cell-free conditioned media (CFCM) samples obtained from Lcb. rhamnosus CRL 2244 cultures incubated at different times display antimicrobial activity against 43 different pathogens, including CRAB, methicillin-resistant Staphylococcus aureus (MRSA) and carbapenemase Klebsiella pneumoniae (KPC)-positive strains. Furthermore, transwell and ultrafiltration analyses together with physical and chemical/biochemical tests showed that Lcb. rhamnosus CRL 2244 secretes a <3 kDa metabolite(s) whose antimicrobial activity is not significantly impaired by mild changes in pH, temperature and various enzymatic treatments. Furthermore, sensitivity and time-kill assays showed that the bactericidal activity of the Lcb. rhamnosus CRL 2244 metabolite(s) enhances the activity of some current FDA approved antibiotics. We hypothesize that this observation could be due to the effects of Lcb. rhamnosus CRL 2244 metabolite(s) on cell morphology and the enhanced transcriptional expression of genes coding for the phenylacetate (PAA) and histidine catabolic Hut pathways, metal acquisition and biofilm formation, all of which are associated with bacterial virulence. Interestingly, the extracellular presence of Lcb. rhamnosus CRL 2244 induced the transcription of the gene coding for the CidA/LgrA protein, which is involved in programmed cell death in some bacteria. Overall, the findings presented in this report underscore the promising potential of the compound(s) released by Lcb. rhamnosus CRL2244 as an alternative and/or complementary option to treat infections caused by A. baumannii as well as other MDR bacterial pathogens. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii is a health threat due to its antibiotic resistance. Herein, antibiotic susceptibility and its association with the Toxin-antitoxin (TA) system genes in A. baumannii clinical isolates from Iran were investigated. Next, we prepared meropenem-loaded chitosan nanoparticles (MP-CS) and investigated their antibacterial effects against meropenem-susceptible bacterial isolates. Methods: Out of 240 clinical specimens, 60 A. baumannii isolates were assessed. Antibiotic resistance of the isolates against conventional antibiotics was determined alongside investigating the presence of three TA system genes (mazEF, relBE, and higBA). Chitosan nanoparticles were characterized in terms of size, zeta potential, encapsulation efficiency, and meropenem release activity. Their antibacterial effects were assessed using the well diffusion method, minimum inhibitory concentration (MIC), and colony-forming unit (CFU) counting. Their cytotoxic effects and biocompatibility index were determined via the MTT, LDH, and ROS formation assays. Results: Ampicillin, ceftazidime, and colistin were the least effective, and amikacin and tobramycin were the most effective antibiotics. Out of the 60 isolates, 10 (16.7%), 5 (8.3%), and 45 (75%) were multidrug-resistant (MDR), extensively drug-resistant (XDR), and pandrug-resistant (PDR), respectively. TA system genes had no significant effect on antibiotic resistance. MP-CS nanoparticles demonstrated an average size of 191.5 and zeta potential of 27.3 mV alongside a maximum encapsulation efficiency of 88.32% and release rate of 69.57%. MP-CS nanoparticles mediated similar antibacterial effects, as compared with free meropenem, against the A. baumannii isolates with significantly lower levels of meropenem. MP-CS nanoparticles remarkably prevented A549 and NCI-H292 cell infection by the A. baumannii isolates alongside demonstrating a favorable biocompatibility index. Conclusion: Antibiotic-loaded nanoparticles should be further designed and investigated to increase their antibacterial effect against A. baumannii and assess their safety and applicability in vivo settings. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a non-fermentative Gram-negative bacterium that can cause nosocomial infections in critically ill patients. Carbapenem-resistant A. baumannii (CRAB) has spread rapidly in clinical settings and has become a key concern. The main objective of this study was to identify the distribution of integrons and biofilm-formation-related virulence genes in CRAB isolates. A total of 269 A. baumannii isolates (219 isolates of CRAB and 50 isolates of carbapenem-sensitive A. baumannii (CSAB)) were collected. Carbapenemase genes (blaKPC, blaVIM, blaIMP, blaNDM, and blaOXA-23-like) and biofilm-formation-related virulence genes (abal, bfms, bap, and cusE) were screened with PCR. Class 1 integron was screened with PCR, and common promoters and gene cassette arrays were determined with restriction pattern analysis combined with primer walking sequencing. Whole-genome sequencing was conducted, and data were analyzed for a blaOXA-23-like-negative isolate. All 219 CRAB isolates were negative for blaKPC, blaVIM, blaIMP, and blaNDM, while blaOXA-23-like was detected in 218 isolates. The detection rates for abal, bfms, bap, and cusE in 219 CRAB were 93.15%, 63.93%, 88.13%, and 77.63%, respectively. Class 1 integron was detected in 75 CRAB (34.25%) and in 3 CSAB. The single gene cassette array aacA4-catB8-aadA1 with relatively strong PcH2 promoter was detected in class 1 integrons. The blaOXA-23-like-negative CRAB isolate was revealed to be a new sequence type (Oxford 3272, Pasteur 2520) carrying blaOXA-72, blaOXA-259, and blaADC-26. In conclusion, blaOXA-23-like was the main reason for CRAB's resistance to carbapenems. A new (Oxford 3272, Pasteur 2520) CRAB sequence type carrying the blaOXA-72, blaOXA-259, and blaADC-26 was reported. [ABSTRACT FROM AUTHOR]

Background: Biofilm development by bacteria is considered to be an essential stage in the bacterial infection. Acinetobacter nosocomialis is an important nosocomial pathogen causing a variety of human infections. However, characteristics and specific determinants of biofilm development have been poorly characterized in A. nosocomialis. Objective: The aim of this study was to investigate the factors involved in the biofilm development by A. nosocomialis. Methods: Library of random transposon mutants was constructed using the Tn5 mutagenesis. The mutant strains, in which the ability of biofilm formation was significantly impaired, were screened by gentian violet staining. The roles of BfmR and BfmS were determined by constructing a bfmR and bfmS deletion mutant and analyzing the effects of bfmR and bfmS mutation on the biofilm development and motility of A. nosocomialis. Results: We identified a biofilm-defective mutant in which a transposon insertion inactivated an open reading frame encoding the BfmR in a two-component regulatory system consisting of BfmR and BfmS. The bfmR mutant revealed a significant reduction in biofilm formation and motility compared to wild-type strain. Deficiency in the biofilm formation and motility of the bfmR mutant was restored by single copy bfmR complementation. In contrast, the bfmS mutant had no effect on biofilm formation. Conclusion: A. nosocomialis has a two-component regulatory system, BfmRS. BfmR is a response regulator required for the initial attachment and maturation of biofilm during the biofilm development as well as the bacterial growth. BfmR could be a potential drug target for A. nosocomialis infection. [ABSTRACT FROM AUTHOR]

Acinetobacter is an important nosocomial pathogen causing health care associated infections. It is highly antibiotic resistant gram-negative bacilli. The study was done to determine the prevalence of Acinetobacter species isolated from various clinical samples with their antibiotic susceptibility pattern. To determine the antimicrobial susceptibility pattern of the isolated Acinetobacter species and also the multidrug resistant mechanisms by phenotypic characterization. The retrospective study was carried out in patients diagnosed with Acinetobacter infection in the Microbiology Department, Krishna Institute of Medical Sciences, Krishna Vishwa Vidyapeeth, Deemed To Be University, Karad, a tertiary care hospital, including the clinical departments, during the period of two years from November 2020 till November 2022. Organism identification, biochemical test, antibiotic resistance pattern and phenotypic tests such as ESBL, MBL and Carbapenemase production were performed as per the Clinical and Laboratory Standards Institute (CLSI). The Modified Hodge test (MHT) was performed for Carbapenemase production detection in Acinetobacter species. A total of 150 Acinetobacter species were isolated from clinical samples. Acinetobacter baumannii was the most prevalent species 138 (92%), followed by Acinetobacter lwoffii 10 (7%) and Acinetobacter hemolyticus 2 (1%). The isolates showed highest resistance to Ampicillin 130 (87%) and sensitive to Colistin 113 (75.3%). Most of the isolates of of Acinetobacter baumannii showed maximum ESBL 21(14%) and MBL 75 (93%) production. Modified Hodge test showed positive results in Acinetobacter baumannii, only 11 (7%). The study showed that Acinetobacter baumannii was the most prevalent species showing drug resistance by phenotypic detection methods. At present Acinetobacter is a frequent pathogen in hospital acquired infections in critically ill patients admitted to ICU. The isolates of Acinetobacter species in our study showed resistant to most commonly used antibiotics. The study showed that ESBL production in Acinetobacter was 22 (15%) and MBL 80 (53%). Most Acinetobacter isolates were Multi Drug Resistant (MDR). MDR Acinetobacter is widely increasing due to inappropriate use of antibiotics in healthcare hospital. In our study, detection of carbapenemase by Modified Hodge test was positive in 11 (7%) isolates. [ABSTRACT FROM AUTHOR]

Objective: The use of essential oils for the preparation of different nanoemulsions (NEa, NEb and NEc) and detect their biological activities. Methods: Nanoemulsiona were prepared by mixing the essential oil (10%) of Coriandrum seed of Coriandrum sativum, Ginger roots of Zingiber officinale, or Achillea leaves of Achillea clavennae), sterile water, and surfactants (Brij 30, Span 20, Triton X-100, Tween 60) in addition to the 0.1M Tris-HCl buffer (pH 7.22). The three prepared nanoemulsions were examined and characterized. Their antibacterial activities were examined against some multidrug-resistant Gram-positive and negative bacteria. Results: The maximum activity was recorded by NEb against Serratia marcescens, Staphylococcus aurous, Micrococcus luteus, Enterococcus faecalis, Streptococcus pyogenes, and Staphylococcus saprophyticus while lower activity was obtained against Klebsiella pneumonia and Escherichia coli. The minimum inhibitory concentrations (MICs) of the NEb ranged from 40--60 µl/ml of the nutrient broth. NEb affects the cell counts and morphology of S. marcescens and destroys the bacterial cells by reducing cell respiration and enhancing the cell permeability and leakage of protein, DNA, and potassium of the bacterial cell membrane. NEb recorded no toxicity against Artimia salina and HaCaT cell line. It decreases biofilm formation by 51%, bacterial infection, and cell attachment. Conclusion: This study suggests that NEs have great antibacterial activity against some human pathogens and process different modes of action, thus they can be used effectively to treat different bacterial infections. [ABSTRACT FROM AUTHOR]

Acinetobacter pneumonia is a significant healthcare-associated infection that poses a considerable challenge to clinicians due to its multidrug-resistant nature. Recent world events, such as the COVID-19 pandemic, have highlighted the need for effective treatment and management strategies for Acinetobacter pneumonia. In this review, we discuss lessons learned from recent world events, particularly the COVID-19 pandemic, in the context of the treatment and management of Acinetobacter pneumonia. We performed an extensive literature review to uncover studies and information pertinent to the topic. The COVID-19 pandemic underscored the importance of infection control measures in healthcare settings, including proper hand hygiene, isolation protocols, and personal protective equipment use, to prevent the spread of multidrug-resistant pathogens like Acinetobacter. Additionally, the pandemic highlighted the crucial role of antimicrobial stewardship programs in optimizing antibiotic use and curbing the emergence of resistance. Advances in diagnostic techniques, such as rapid molecular testing, have also proven valuable in identifying Acinetobacter infections promptly. Furthermore, due to the limited availability of antibiotics for treating infections caused A. baumannii, alternative strategies are needed like the use of antimicrobial peptides, bacteriophages and their enzymes, nanoparticles, photodynamic and chelate therapy. Recent world events, particularly the COVID-19 pandemic, have provided valuable insights into the treatment and management of Acinetobacter pneumonia. These lessons emphasize the significance of infection control, antimicrobial stewardship, and early diagnostics in combating this challenging infection. [ABSTRACT FROM AUTHOR]

Introduction: Biofilm formation is an important virulence factor of Acinetobacter baumannii. Here, we examined the biofilm formation of archived A. baumannii causing ventilator‐associated pneumonia (VAP). Methods: Eighteen and twenty isolates of A. baumannii causing bacteremic pneumonia and non‐bacteremic pneumonia were included, respectively. Antimicrobial susceptibility testing was performed by broth microdilution method, while biofilm formation was evaluated by microtiter dish biofilm formation assay. Results: All 38 isolates were still susceptible to colistin and tigecycline, whereas almost all isolates were non‐susceptible (intermediate to resistant) to several antimicrobial agents, especially ceftriaxone and cefotaxime. Approximately, 44% of bacteremic isolates and 50% of non‐bacteremic isolates were classified as carbapenem‐resistant A. baumannii (CRAB). Biofilm formation was detected in 42% of the studied isolates. Bacteremia among the patients infected with biofilm‐producing isolates was significantly higher than in those infected with non‐biofilm‐producing isolates. The antimicrobial susceptibilities of A. baumannii with biofilm formation were lower than those without biofilm formation, but the differences did not have statistical significance. The patients infected with non‐biofilm‐producing isolates had good clinical and non‐clinical outcomes than those infected with biofilm‐producing isolates. The survival rate of patients diagnosed with VAP due to biofilm‐producing A. baumannii was lower than in those patients diagnosed with VAP due to non‐biofilm‐producing isolates. Conclusion: Biofilm formation of A. baumannii causing VAP was associated with antimicrobial resistance and bacteremia as well as unfavorable clinical outcomes. [ABSTRACT FROM AUTHOR]

Methylotrophs are a diverse group of bacteria that abundantly colonize the phyllosphere and have great potential to withstand UV irradiation because of their pigmented nature and ability to promote plant growth through various mechanisms. The present study investigated the effects of UVB radiation on plant growth-promoting (PGP) properties of methylotrophic bacteria and the growth of Vigna radiata L. A total of 55 methylotrophic bacteria were isolated from desert plants, and 15 methylotrophs were resistant to UVB radiation for 4 h. All UVB-resistant methylotrophs possess a methyldehydrogenase gene. Identification based on 16S rRNA gene sequencing revealed that all 15 UVB-resistant methylotrophs belonged to the genera Methylorubrum (07), Methylobacterium (07), and Rhodococcus (01). Screening of methylotrophs for PGP activity in the presence and absence of UVB radiation revealed that all isolates showed ACC deaminase activity and growth on a nitrogen-free medium. Furthermore, the production of IAA-like substances ranged from 8.62 to 85.76 µg/mL, siderophore production increased from 3.47 to 65.75% compared to the control. Seed germination assay with V. radiata L. (mung bean) exposed to UVB radiation revealed that methylotrophs improved seed germination, root length, and shoot length compared to the control. The present findings revealed that the isolates SD3, SD2, KD1, KD5, UK1, and UK3 reduced the deleterious effects of UVB radiation on mung bean plants and can be used to protect seedlings from UVB radiation for sustainable agriculture. [ABSTRACT FROM AUTHOR]

Introduction: Acinetobacter spp. has emerged as an important hospital-acquired and opportunistic agent due to its survival capability in adverse conditions, saprophytic presence, and increasing resistance to antimicrobials. The irrational use of antibiotics, along with biofilm formation, plays an important role in producing Extensively Drug-resistant (XDR) and Multidrug-Resistant (MDR) Acinetobacter species, especially Acinetobacter calcoaceticus-baumannii complex (Acb complex), in the hospital environment, contributing to morbidity and mortality. Aim: To detect biofilm production among Acinetobacter species isolated from various clinical samples received from Intensive Care Units (ICUs) as well as their antibiotic sensitivity pattern. Materials and Methods: A cross-sectional study was conducted in the Microbiology Department at Teerthanker Mahaveer Medical College and Research Centre, Moradabad, Uttar Pradesh, India from January 2022 to April 2023. Patients of all age groups and both genders were included in the study after obtaining informed consent. Clinical specimens, including endotracheal secretions, endotracheal tips, pus, urine, sputum, tissue, and other body fluids, were collected from ICUs where Acinetobacter spp. was detected. A total of 223 cases were included. The specimens were collected using clean, leak-proof, sterile containers with proper aseptic precautions. Antimicrobial susceptibility testing was performed according to the guidelines set by the Clinical and Laboratory Standards Institute (CLSI) in 2022 and identifies MDR and XDR strains. The biofilm production of isolates was determined using a quantitative adherence assay. The data generated was entered into Microsoft excel, and statistical analysis was conducted using International Business Machines (IBM) Statistical Package for Social Sciences (SPSS) statistical software version 28.0. The results were then presented as descriptive statistics. Results: Among the 223 Acinetobacter spp. isolates, 159 (71.3%) were identified as Acb complex (Acinetobacter calcoaceticus-baumannii complex), followed by A. lwofi with 39 (17.5%) isolates and A. haemolyticus with 16 (7.2%) isolates. Acinetobacter showed resistance, in descending order of frequency, to amoxicillin+clavulanic acid (211 isolates, 94.6%), ciprofloxacin (211 isolates, 94.6%), trimethoprim-sulfamethoxazole (208 isolates, 93.3%), cefotaxime (198 isolates, 88.8%), cefepime (187 isolates, 83.8%), gentamycin (178 isolates, 79.8%), amikacin (152 isolates, 68.2%), piperacillin-tazobactam (68 isolates, 30.5%), imipenem (72 isolates, 32.3%), meropenem (71 isolates, 31.8%), polymyxin B (12 isolates, 5.4%), and colistin (2 isolates, 0.9%). The maximum antibiotic resistance was observed in Acb complex, with 208 (93.3%) strains being MDR producers and 32 (14.3%) strains being XDR producers. Biofilm production was observed in 214 isolates (95.9%), with 127 (56.9%) exhibiting strong biofilm production 63 (28.2%) showing moderate biofilm production, and 24 (10.8%) showing weak biofilm production. All MDR strains were found to produce biofilm, and out of those, 127 (61.1%) exhibited strong biofilm production. Among the XDR strains, all 32 (100%) were found to produce strong biofilm. Conclusion: In conclusion, Acinetobacter spp. has a high propensity for developing MDR, and the formation of biofilms further aids the organism in surviving under strenuous conditions, making it difficult to treat. Therefore, regular surveillance of Hospital-acquired Infections (HAI), the prevention of misuse and overuse of antibiotics, prescribing antibiotics based on antibiogram patterns, formulating antibiotic policies, and implementing bundle care approaches for the prevention of HAI are crucial in preventing antibiotic resistance. [ABSTRACT FROM AUTHOR]

Background: Multidrug-resistant Acinetobacter baumannii rarely causes community-acquired pneumonia. Here, we report the clinical and genomic characteristics of a multidrug-resistant A. baumannii strain responsible for community-acquired pneumonia in a 31-year-old healthy young women.Methods:A. baumannii strain W2LL was recovered from the alveolar lavage fluid sample of a hospitalized patient with pulmonary infection. Growth rate studies were conducted under various conditions, and virulence assessments were performed using Galleria Mellonella larvae. Whole Genome Sequencing (WGS) was carried out using Oxford Nanopore MinIon and Illumina HiSeq. In silico multilocus sequence typing (MLST), plasmid replicons, antimicrobial resistance genes, and virulence genes were determined using the BacWGSTdb webserver. Phylogenetic analysis between strain W2LL and other closely related A. baumannii genomes retrieved from NCBI database was performed.Results: WGS identified strain W2LL as a rare sporadic lineage sequence type (ST) 1431. In addition to the detection of the β-lactamase gene (blaOXA-98) on the chromosome, blaOXA-58 was found on a 92,034 bp plasmid. Antimicrobial susceptibility testing revealed this strain was resistant to cephalosporins and carbapenems, with initial treatment using cefoxitin proving ineffective. Subsequent treatment with piperacillin-sulbactam combined with levofloxacin led to gradual improvement. Compared to A. baumannii ATCC 17978, W2LL exhibited similar growth rates at 37°C and 42°C, as well as in the presence of zinc. However, strain W2LL exhibited higher virulence phenotype compared to ATCC 17978 in G. mellonella model. The closest relative of A. baumannii W2LL was CAM180_1, another isolate recovered from Cambodia, which differed by 191 SNPs.Conclusion: W2LL is a rare ST1431 carbapenem-resistant A. baumannii strain recovered from a patient with no prior hospitalization or typical risk factors. This underscores the growing menace posed by carbapenem-resistant A. baumannii, no longer limited to hospitalized patients, potentially impacting the broader, younger population. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii, the first human pathogen to be designated as a "red-alert" pathogen, is on the critical priority list of pathogens requiring new antibiotics. Biofilm-associated diseases are the most common infections caused by the antibiotic-resistant bacteria A. baumannii. Multidrug-resistant strains are more easily transmitted around the world due to A. baumannii's ability to produce biofilms, which allows it to develop antibiotic resistance mechanisms and thrive in healthcare environments. As a result, A. baumannii infections are becoming increasingly common in hospital settings allover the world. As a result, a comprehensive systematic review and meta-analysis were carried out to determine the global prevalence of biofilm-producing A. baumannii clinical isolates. Methods: Articles were extensively searched in bibliographic databases and grey literatures using entry terms or phrases. Studies meeting eligibility criteria were extracted in MS Excel and exported into STATA version 12 software for statistical analysis. A random-effects model was used to compute the pooled prevalence of biofilm-producing A. baumannii clinical isolates. The heterogeneity was quantified by using the I2 value. Publication bias was assessed using a funnel plot and Egger's test. Sensitivity analysis was done to assess the impact of a single study on pooled effect size. Result: Of the 862 studies identified, 26 studies consisted of 2123 A.baumannii clinical isolates of which 1456 were biofilm-producing. The pooled prevalence of biofilm-producing A.baumannii clinical isolates was 65.63% (95% CI = 56.70%-74.56%). There was substantial heterogeneity with an I2 value of 98.1%. Moreover, 41.34%, 33.57%, and 27.63% of isolates of strong, mild, and weak producers of biofilm. Higher prevalence was found in studies published after 2014 (66.31%); Western Pacific region (76.17%); and Asia (66.22%) followed by the African continent (57.29%). Conclusion: The pooled prevalence of biofilm-producing A. baumannii clinical isolates has risen alarmingly, posing a public health risk. This indicates the burden of biofilm-producing A. baumannii infections urges routine screening and appropriate treatment for better management of hospitalized patients, as well as effective controlling of the emergence of drug resistance. Furthermore, this finding is an alert call for the stakeholders to develop strong infection prevention and antibiotics stewardship programs for the prevention and control of biofilm-producing bacterial infections. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii (A. baumannii) is an opportunistic pathogen that poses dangerous health threat. It is a main cause of biofilm-associated infections that are mostly resistant to antibiotic therapy. Because of its capacity to form biofilm on biotic and abiotic surfaces, it has been linked to most nosocomial infections such as ventilator-associated pneumonia, urinary tract infections, bacteremia, meningitis, wound infections, soft tissue infections, and peritonitis. The biofilm refers to an organized group of microbial cells that are embedded in an exopolymeric substance made of protein, extracellular DNA, and polysaccharide. Bacterial cells in biofilms are resistant to chemicals, phagocytosis, and other elements of the body's innate and acquired immune systems posing treatment challenges. Biofilm formation in A. baumannii is a complicated process that is influenced by a variety of factors such as outer membrane protein A, poly-β-(1,6)-N acetyl glucosamine (PAGE), biofilm-associated protein, two-component system (Bfm/S BfmR), chaperone–usher (Csu) pilus assembly system of pili, BlaPER-1 belonging to β-lactamase family, extracellular polymeric substance, and the quorum sensing system. Several biofilm-associated genes influence antimicrobial susceptibility, implying a link between biofilm formation and antimicrobial resistance. Short conclusion: This review describes the complex biofilm system of A. baumannii, which gives it a survival advantage and increases its colonization ability. Also, it demonstrates various extrinsic and intrinsic factors that function and regulate the biofilm machinery of A. baumannii. Furthermore, this study considers prospective ways for preventing biofilm development on relevant medical equipment, as well as potential therapeutic strategies for eradicating mature biofilms, which can aid in the treatment of biofilm-associated A. baumannii infection. [ABSTRACT FROM AUTHOR]

It is now clearly recognized that light modulates the physiology of many bacterial chemotrophs, either directly or indirectly. An interesting case are bacterial pathogens of clinical relevance. This work summarizes, discusses, and provides novel complementary information to what is currently known about light sensing and responses in critical human pathogens such as Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus aureus. These pathogens are associated with severe hospital and community infections difficult to treat due to resistance to multiple drugs. Moreover, light responses in Brucella abortus, an important animal and human pathogen, are also compiled. Evidence recovered so far indicates that light modulates aspects related to pathogenesis, persistence, and antibiotic susceptibility in these pathogens; such as motility, biofilm formation, iron uptake, tolerance to antibiotics, hemolysis and virulence. The pathogens elicit differential responses to light depending likely on their pathophysiology, ability to cause disease and characteristics of the host. The response to light is not restricted to discrete physiological traits but is global. In higher organisms, light provides spatial and temporal information. Then, it is crucial to understand what information light is providing in these bacterial pathogens. Our current hypothesis postulates that light serves as a signal that allows these pathogens to synchronize their behavior to the circadian rhythm of the host, to optimize infection. Advances on the molecular mechanism of light signal transduction and physiological responses to light, as well as in the relation between light and bacterial infection, would not only enlarge our understanding of bacterial pathogenesis but also could potentially provide alternative treatment options for infectious illnesses. [ABSTRACT FROM AUTHOR]

Carbapenem-resistant Acinetobacter baumannii (CRAB) is a recognized nosocomial pathogen with limited antibiotic treatment options. Lactic acid bacteria (LAB) constitute a promising therapeutic alternative. Here we studied the antibacterial properties of a collection of LAB strains using phenotypic and transcriptomic analysis against A. baumannii clinical strains. One strain, Lacticaseibacillus rhamnosus CRL 2244, demonstrated a potent inhibitory capacity on A. baumannii with a significant killing activity. Scanning electron microscopy images showed changes in the morphology of A. baumannii with an increased formation of outer membrane vesicles. Significant changes in the expression levels of a wide variety of genes were also observed. Interestingly, most of the modified genes were involved in a metabolic pathway known to be associated with the survival of A. baumannii. The paa operon, Hut system, and fatty acid degradation were some of the pathways that were induced. The analysis reveals the impact of Lcb. rhamnosus CRL 2244 on A. baumannii response, resulting in bacterial stress and subsequent cell death. These findings highlight the antibacterial properties of Lcb. rhamnosus CRL 2244 and its potential as an alternative or complementary strategy for treating infections. Further exploration and development of LAB as a treatment option could provide valuable alternatives for combating CRAB infections. [ABSTRACT FROM AUTHOR]

Plant–pathogen interaction is influenced by multiple environmental factors, including temperature and light. Recent works have shown that light modulates not only the defense response of plants but also the pathogens virulence. Xanthomonas citri subsp. citri (Xcc) is the bacterium responsible for citrus canker, an important plant disease worldwide. The Xcc genome presents four genes encoding putative photoreceptors: one bacteriophytochrome and three blue light photoreceptors, one LOV and two BLUFs (bluf1: XAC2120 and bluf2: XAC3278). The presence of two BLUFs proteins is an outstanding feature of Xcc. In this work we show that the bluf2 gene is functional. The mutant strain, XccΔbluf2, was constructed demonstrating that BLUF2 regulates swimming-type motility, adhesion to leaves, exopolysaccharide production and biofilm formation, features involved in the Xcc virulence processes. An important aspect during the plant–pathogen interaction is the oxidative response of the host and the consequent reaction of the pathogen. We observed that ROS detoxification is regulated by Xcc bluf2 gene. The phenotypes of disease in orange plants produced by WT and XccΔbluf2 strains were evaluated, observing different phenotypes. Altogether, these results show that BLUF2 negatively regulates virulence during citrus canker. This work constitutes the first report on BLUF-like receptors in plant pathogenic bacteria. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the antibiotic resistance and biofilm formation of Acinetobacter calcoaceticus—A. baumannii (ACB) complex isolates recovered from a university hospital in Pelotas, RS, Brazil. The species were confirmed using gyrB multiplex and blaOXA-51-like genes PCR. The presence of the bfmRS virulence gene was evaluated by the PCR, and the isolates were classified based on their biofilm-forming ability on polystyrene (PO) and glass surfaces (TM). Out of 50 ACB complex isolates evaluated, 41 were identified as A. baumannii and nine as A. nosocomialis. The bfmRS gene was detected in 97.6% (40/41) of A. baumannii and 33.3% (3/9) of A. nosocomialis species. Forty-nine isolates exhibited a multidrug-resistant (MDR) profile, while one A. nosocomialis isolate presented an extensively drug-resistant (XDR) profile. All isolates were able of forming biofilms on PO surfaces and 98% (49/50) on TM surfaces. A significant correlation was observed between biofilm production on PO and TM surfaces (P < 0.05). However, no correlation was found between biofilms forming and the presence of the bfmRS gene or displaying a certain antibiotic resistance profile. In conclusion, A. baumannii and A. nosocomialis are frequent species causing nosocomial infections in a hospital in Pelotas, RS, Brazil, and both are capable of forming biofilms. [ABSTRACT FROM AUTHOR]

Biofilm-related infections substantially contribute to bacterial illnesses, with estimates indicating that at least 80% of such diseases are linked to biofilms. Biofilms exhibit unique metabolic patterns that set them apart from their planktonic counterparts, resulting in significant metabolic reprogramming during biofilm formation. Differential glycolytic enzymes suggest that central metabolic processes are markedly different in biofilms and planktonic cells. The glycolytic enzyme glyceraldehyde-3-phosphate dehydrogenase (GAPDH) is highly expressed in Staphylococcus aureus biofilm progenitors, indicating that changes in glycolysis activity play a role in biofilm development. Notably, an important consideration is a correlation between elevated cyclic di-guanylate monophosphate (c-di-GMP) activity and biofilm formation in various bacteria. C-di-GMP plays a critical role in maintaining the persistence of Pseudomonas aeruginosa biofilms by regulating alginate production, a significant biofilm matrix component. Furthermore, it has been demonstrated that S. aureus biofilm development is initiated by several tricarboxylic acid (TCA) intermediates in a FnbA-dependent manner. Finally, Glucose 6-phosphatase (G6P) boosts the phosphorylation of histidine-containing protein (HPr) by increasing the activity of HPr kinase, enhancing its interaction with CcpA, and resulting in biofilm development through polysaccharide intercellular adhesion (PIA) accumulation and icaADBC transcription. Therefore, studying the metabolic changes associated with biofilm development is crucial for understanding the complex mechanisms involved in biofilm formation and identifying potential targets for intervention. Accordingly, this review aims to provide a comprehensive overview of recent advances in metabolomic profiling of biofilms, including emerging trends, prevailing challenges, and the identification of potential targets for anti-biofilm strategies. [ABSTRACT FROM AUTHOR]

The Gram-negative bacterial pathogen Acinetobacter baumannii is a major cause of hospital-acquired opportunistic infections. The increasing spread of pan-drug resistant strains makes A. baumannii top-ranking among the ESKAPE pathogens for which novel routes of treatment are urgently needed. Comparative genomics approaches have successfully identified genetic changes coinciding with the emergence of pathogenicity in Acinetobacter. Genes that are prevalent both in pathogenic and a-pathogenic Acinetobacter species were not considered ignoring that virulence factors may emerge by the modification of evolutionarily old and widespread proteins. Here, we increased the resolution of comparative genomics analyses to also include lineage-specific changes in protein feature architectures. Using type IVa pili (T4aP) as an example, we show that three pilus components, among them the pilus tip adhesin ComC, vary in their Pfam domain annotation within the genus Acinetobacter. In most pathogenic Acinetobacter isolates, ComC displays a von Willebrand Factor type A domain harboring a finger-like protrusion, and we provide experimental evidence that this finger conveys virulence-related functions in A. baumannii. All three genes are part of an evolutionary cassette, which has been replaced at least twice during A. baumannii diversification. The resulting strain-specific differences in T4aP layout suggests differences in the way how individual strains interact with their host. Our study underpins the hypothesis that A. baumannii uses T4aP for host infection as it was shown previously for other pathogens. It also indicates that many more functional complexes may exist whose precise functions have been adjusted by modifying individual components on the domain level. Author summary: Type IVa pili (T4aP) are hair-like, extendable, and retractable appendages that many bacteria use for interacting with their environment. Several human pathogens have independently recruited these pili for processes related to host infection, but the modifications necessary to turn T4aP into virulence factors are largely unknown. Here, we studied if and how T4aP components have changed in the nosocomial pathogen A. baumannii compared to its largely a-pathogenic relatives in the Acinetobacter genus. Most A. baumannii isolates have T4aP with a pilus tip adhesin containing a protein domain variant not seen outside the pathogenic clade. This variant appears essential for bacterial motility and contributes to host cell adhesion and natural competence. However, some isolates have T4aP resembling those of largely a-pathogenic species in this genus. This indicates that the way these pili are used during infection processes differs between A. baumannii isolates probably as a consequence of niche adaptation. In a broader perspective, our findings highlight that many relevant genetic differences between pathogens and their a-pathogenic relatives emerge only on the domain- and sub-domain level. Thus, existing comparative genomics studies have likely uncovered only the tip of the iceberg of genetic determinants that contribute to A. baumannii virulence. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a Gram-negative bacterial pathogen that exhibits high intrinsic resistance to antimicrobials, with treatment often requiring the use of last-resort antibiotics. Antibiotic-resistant strains have become increasingly prevalent, underscoring a need for new therapeutic interventions. The aim of this study was to use A. baumannii outer membrane vesicles as immunogens to generate single-domain antibodies (VHHs) against bacterial cell surface targets. Llama immunization with the outer membrane vesicle preparations from four A. baumannii strains (ATCC 19606, ATCC 17961, ATCC 17975, and LAC-4) elicited a strong heavy-chain IgG response, and VHHs were selected against cell surface and/or extracellular targets. For one VHH, OMV81, the target antigen was identified using a combination of gel electrophoresis, mass spectrometry, and binding studies. Using these techniques, OMV81 was shown to specifically recognize CsuA/B, a protein subunit of the Csu pilus, with an equilibrium dissociation constant of 17 nM. OMV81 specifically bound to intact A. baumannii cells, highlighting its potential use as a targeting agent. We anticipate the ability to generate antigen-specific antibodies against cell surface A. baumannii targets could provide tools for further study and treatment of this pathogen. Key points: •Llama immunization with bacterial OMV preparations for VHH generation •A. baumannii CsuA/B, a pilus subunit, identified by mass spectrometry as VHH target •High-affinity and specific VHH binding to CsuA/B and A. baumannii cells [ABSTRACT FROM AUTHOR]

The Acinetobacter baumannii bacteriophage Aci01-1, which belongs to the genus Saclayvirus of the order Caudoviricetes, has an icosahedral head and a contractile rigid tail. We report that Aci01-1 has, attached to the tail conical tip, a remarkable 146-nm-long flexible fiber with seven beads and a terminal knot. Its putative gene coding for a 241.36-kDa tail fiber protein is homologous to genes in Aci01-1-related and unrelated phages. Analysis of its 3D structure using AlphaFold provides a structural model for the fiber observed by electron microscopy. We also identified a putative receptor of the phage on the bacterial capsule that is hypothesized to interact with the Aci01-1 long fiber. [ABSTRACT FROM AUTHOR]

Motivation Wastewater treatment plants (WWTPs) harbor a dense and diverse microbial community. They constantly receive antimicrobial residues and resistant strains, and therefore provide conditions for horizontal gene transfer (HGT) of antimicrobial resistance (AMR) determinants. This facilitates the transmission of clinically important genes between, e.g. enteric and environmental bacteria, and vice versa. Despite the clinical importance, tools for predicting HGT remain underdeveloped. Results In this study, we examined to which extent water cycle microbial community composition, as inferred by partial 16S rRNA gene sequences, can predict plasmid permissiveness, i.e. the ability of cells to receive a plasmid through conjugation, based on data from standardized filter mating assays using fluorescent bio-reporter plasmids. We leveraged a range of machine learning models for predicting the permissiveness for each taxon in the community, representing the range of hosts a plasmid is able to transfer to, for three broad host-range resistance IncP plasmids (pKJK5, pB10, and RP4). Our results indicate that the predicted permissiveness from the best performing model (random forest) showed a moderate-to-strong average correlation of 0.49 for pB10 [95% confidence interval (CI): 0.44–0.55], 0.43 for pKJK5 (0.95% CI: 0.41–0.49), and 0.53 for RP4 (0.95% CI: 0.48–0.57) with the experimental permissiveness in the unseen test dataset. Predictive phylogenetic signals occurred despite the broad host-range nature of these plasmids. Our results provide a framework that contributes to the assessment of the risk of AMR pollution in wastewater systems. Availability and implementation The predictive tool is available as an application at https://github.com/DaneshMoradigaravand/PlasmidPerm. [ABSTRACT FROM AUTHOR]

Background: The gut microbiota plays an important role in human health, as it can affect host immunity and susceptibility to infectious diseases. Invasive intestinal candidiasis is strongly associated with gut microbiota homeostasis. However, the nature of the interaction between Candida albicans and gut bacteria remains unclear. Objective: This review aimed to determine the nature of interaction and the effects of gut bacteria on C. albicans so as to comprehend an approach to reducing intestinal invasive infection by C. albicans. Methods: This review examined 11 common gut bacteria's interactions with C. albicans, including Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecalis, Staphylococcus aureus, Salmonella spp., Helicobacter pylori, Lactobacillus spp., Bacteroides spp., Clostridium difficile, and Streptococcus spp. Results: Most of the studied bacteria demonstrated both synergistic and antagonistic effects with C. albicans, and just a few bacteria such as P. aeruginosa, Salmonella spp., and Lactobacillus spp. demonstrated only antagonism against C. albicans. Conclusions: Based on the nature of interactions reported so far by the literature between gut bacteria and C. albicans, it is expected to provide new ideas for the prevention and treatment of invasive intestinal candidiasis. [ABSTRACT FROM AUTHOR]

Bacterial pneumonia is a common infection of the lower respiratory tract that can afflict patients of all ages. Multidrug-resistant strains of Acinetobacter baumannii are increasingly responsible for causing nosocomial pneumonias, thus posing an urgent threat. Alveolar macrophages play a critical role in overcoming respiratory infections caused by this pathogen. Recently, we and others have shown that new clinical isolates of A. baumannii, but not the common lab strain ATCC 19606 (19606), can persist and replicate in macrophages within spacious vacuoles that we called AcinetobacterContaining Vacuoles (ACV). In this work, we demonstrate that the modern A. baumannii clinical isolate 398, but not the lab strain 19606, can infect alveolar macrophages and produce ACVs in vivo in a murine pneumonia model. Both strains initially interact with the alveolar macrophage endocytic pathway, as indicated by EEA1 and LAMP1 markers; however, the fate of these strains diverges at a later stage. While 19606 is eliminated in an autophagy pathway, 398 replicates in ACVs and are not degraded. We show that 398 reverts the natural acidification of the phagosome by secreting large amounts of ammonia, a by-product of amino acid catabolism. We propose that this ability to survive within macrophages may be critical for the persistence of clinical A. baumannii isolates in the lung during a respiratory infection. Author summary: Acinetobacter baumannii is an important nosocomial pathogen with the highest rate of multi-drug resistance among Gram negative bacteria. Although A. baumannii is capable to cause multiple types of infection, pneumonia is the main manifestation of this pathogen. We have recently shown that recent A. baumannii clinical isolates are able to multiply and persist in macrophages in vitro. Here we show that in a murine respiratory infection model, A. baumannii can also create niches within resident macrophages of the lung. Furthermore, we demonstrate that A. baumannii manipulates the macrophage endocytic pathway to create a replicative niche. Recent clinical isolates can withstand the acidic environment on the vacuole and neutralize it by secreting ammonia to the vacuolar lumen, which allows A. baumannii to further multiply inside the macrophage. The ability to survive and multiply within macrophages could be important to avoid the host immune system, avoid clearance, and withstand antibiotic treatment as many antibiotics cannot penetrate host cells. [ABSTRACT FROM AUTHOR]

Phage have gained renewed interest as an adjunctive treatment for life-threatening infections with the resistant nosocomial pathogen Acinetobacter baumannii. Our understanding of how A. baumannii defends against phage remains limited, although this information could lead to improved antimicrobial therapies. To address this problem, we identified genome-wide determinants of phage susceptibility in A. baumannii using Tn-seq. These studies focused on the lytic phage Loki, which targets Acinetobacter by unknown mechanisms. We identified 41 candidate loci that increase susceptibility to Loki when disrupted, and 10 that decrease susceptibility. Combined with spontaneous resistance mapping, our results support the model that Loki uses the K3 capsule as an essential receptor, and that capsule modulation provides A. baumannii with strategies to control vulnerability to phage. A key center of this control is transcriptional regulation of capsule synthesis and phage virulence by the global regulator BfmRS. Mutations hyperactivating BfmRS simultaneously increase capsule levels, Loki adsorption, Loki replication, and host killing, while BfmRS-inactivating mutations have the opposite effect, reducing capsule and blocking Loki infection. We identified novel BfmRS-activating mutations, including knockouts of a T2 RNase protein and the disulfide formation enzyme DsbA, that hypersensitize bacteria to phage challenge. We further found that mutation of a glycosyltransferase known to alter capsule structure and bacterial virulence can also cause complete phage resistance. Finally, additional factors including lipooligosaccharide and Lon protease act independently of capsule modulation to interfere with Loki infection. This work demonstrates that regulatory and structural modulation of capsule, known to alter A. baumannii virulence, is also a major determinant of susceptibility to phage. Author summary: Antibiotic-resistant infections with Acinetobacter baumannii are a major problem in critical care units and have increased in frequency during the COVID-19 pandemic. The virulence of these infections depends on a polysaccharide capsule surrounding the bacterium. Phage, or viruses that kill bacteria, represent a promising alternative therapy against highly antibiotic-resistant A. baumannii infections, and A. baumannii-specific phage often target the capsule. Here, we use high-throughput genetics to analyze how A. baumannii defends against phage and identify ways to potentiate their killing activity. We found that stressing the bacteria in ways that cause augmented production of capsule also causes hyper-susceptibility to phage. By contrast, turning off the stress response, or mutating the capsule structure, causes complete phage resistance. Altering another surface structure, lipooligosaccharide, or an intracellular protease also enhances phage attack. Modulating the amounts or makeup of capsular polysaccharide is known to influence virulence in A. baumannii. This work thus uncovers a connection between phage pressure and the evolution of virulence in A. baumannii, and it identifies control mechanisms that may be leveraged for improving future phage-based antimicrobial therapies. [ABSTRACT FROM AUTHOR]

Acinetobacter species is currently ranked as high-priority pathogen for their extraordinary ability to become resistant to almost all existing antibiotics. The diverse range of effectors secreted by Acinetobacter spp. constitutes a significant proportion of the virulence arsenal. Therefore, our study aims to characterize the secretome of Acinetobacter pittii S-30. Analysis of extracellular secreted proteins of A. pittii S-30 revealed the presence of transporter proteins, outer membrane proteins, molecular chaperones, porins, and several proteins of unknown function. Additionally, proteins related to metabolism, as well as those involved in gene expression and protein translation, type VI secretion system (T6SS) proteins, and stress response-related proteins were also identified in the secretome. The comprehensive analysis of secretome revealed putative protein antigens which could elicit substantial immune response. The limited availability of effective antibiotics and the worldwide growth of secretome data make this approach appealing in the development of effective vaccines against Acinetobacter and other bacterial pathogens. [ABSTRACT FROM AUTHOR]

Background: Acinetobacter baumannii is one of the main causes of healthcare-associated infections that threaten public health, and carbapenems, such as meropenem, have been a therapeutic option for these infections. Therapeutic failure is mainly due to the antimicrobial resistance of A. baumannii, as well as the presence of persister cells. Persisters constitute a fraction of the bacterial population that present a transient phenotype capable of tolerating supra-lethal concentrations of antibiotics. Some proteins have been suggested to be involved in the onset and/or maintenance of this phenotype. Thus, we investigated the mRNA levels of the adeB (AdeABC efflux pump component), ompA, and ompW (outer membrane proteins) in A. baumannii cells before and after exposure to meropenem. Results: We found a significant increase (p-value < 0.05) in the expression of ompA (> 5.5-fold) and ompW (> 10.5-fold) in persisters. However, adeB did not show significantly different expression levels when comparing treated and untreated cells. Therefore, we suggest that these outer membrane proteins, especially OmpW, could be part of the mechanism of A. baumannii persisters to deal with the presence of high doses of meropenem. We also observed in the Galleria mellonella larvae model that persister cells are more virulent than regular ones, as evidenced by their LD50 values. Conclusions: Taken together, these data contribute to the understanding of the phenotypic features of A. baumannii persisters and their relation to virulence, as well as highlight OmpW and OmpA as potential targets for drug development against A. baumannii persisters. [ABSTRACT FROM AUTHOR]

Chaperone-Usher Pathway (CUP) pili are major adhesins in Gram-negative bacteria, mediating bacterial adherence to biotic and abiotic surfaces. While classical CUP pili have been extensively characterized, little is known about so-called archaic CUP pili, which are phylogenetically widespread and promote biofilm formation by several human pathogens. In this study, we present the electron cryomicroscopy structure of the archaic CupE pilus from the opportunistic human pathogen Pseudomonas aeruginosa. We show that CupE1 subunits within the pilus are arranged in a zigzag architecture, containing an N-terminal donor β-strand extending from each subunit into the next, where it is anchored by hydrophobic interactions, with comparatively weaker interactions at the rest of the inter-subunit interface. Imaging CupE pili on the surface of P. aeruginosa cells using electron cryotomography shows that CupE pili adopt variable curvatures in response to their environment, which might facilitate their role in promoting cellular attachment. Finally, bioinformatic analysis shows the widespread abundance of cupE genes in isolates of P. aeruginosa and the co-occurrence of cupE with other cup clusters, suggesting interdependence of cup pili in regulating bacterial adherence within biofilms. Taken together, our study provides insights into the architecture of archaic CUP pili, providing a structural basis for understanding their role in promoting cellular adhesion and biofilm formation in P. aeruginosa. Author summary: Many bacteria adhere to surfaces or host cells using filamentous structures termed pili that extend from the bacterial cell and anchor them to their target. Previous studies have characterised various Chaperone-Usher Pathway (CUP) pili, which are common in Gram-negative bacteria. However, little is known about the so-called archaic CUP pili, which are the most widespread type. Archaic CUP pili help maintain the architecture of multicellular bacterial aggregates termed biofilms formed by the pathogen Pseudomonas aeruginosa and many others. In this study, we present a cryo-EM structure of the archaic CUP pilus CupE from P. aeruginosa, providing a structural basis of how the CupE1 protein forms zigzag-shaped, extended pili. By imaging CupE pili on P. aeruginosa cells using electron cryotomography, we show that pili can adopt variable long-range curvature, which may help their role in providing cohesion between cells within the biofilm. Furthermore, structural modelling provides insights into the roles of minor pilin subunits encoded within the cupE operon. These results will help advance our understanding of bacterial pili structure and function. [ABSTRACT FROM AUTHOR]

Background: Biofilm-associated protein (Bap) in Acinetobacter baumannii is an essential factor in biofilm production and persistence in the hospital environment. Objectives: This study aimed to detect the bap gene in A. baumannii by real-time polymerase chain reaction (PCR) from clinical specimens in Khorramabad, Iran. Methods: This cross-sectional study was performed during April 2017 - April 2018 on 43 A. baumannii strains from clinical samples collected and identified by microbiological and biochemical tests. The expression of the bap gene was evaluated by real-time PCR. Data were analyzed with SPSS version 24. Results: Out of 43 A. baumannii strains, 23, 8, 3, 3, 3, and 3 samples were isolated from chest sputum, wounds, urine, tissues, blood, and, respectively. According to the PCR results, all isolates had the bap gene except one. Real-time PCR showed significant differences in the expression of the bap gene between A. baumannii isolates from diverse clinical samples. The highest expression of the bap gene was found in chest sputum and wound samples and had a significant difference with other samples (P < 0.0001). Conclusions:We observed the bap gene in most strains, with the high expression of this gene in chest sputum and wound samples. Therefore, further studies are recommended to find strategies to inhibit the expression of this gene and biofilm formation, which help treat infections caused by biofilm-forming A. baumannii strains. [ABSTRACT FROM AUTHOR]

Here, we present the genomic characterization of an Acinetobacter bohemicus strain QAC-21b which was isolated in the presence of a quaternary alky-ammonium compound (QAAC) from manure of a conventional German pig farm. The genetic determinants for QAAC, heavy metal and antibiotic resistances are reported based of the whole genome shotgun sequence and physiological growth tests. A. bohemicus QAC-21b grew in a species typical manner well at environmental temperatures but not at 37 °C. The strain showed tolerance to QAACs and copper but was susceptible to antibiotics relevant for Acinetobacter treatments. The genome of QAC-21b contained several Acinetobacter typical QAAC and heavy metal transporting efflux pumps coding genes, but no key genes for acquired antimicrobial resistances. The high genomic content of transferable genetic elements indicates that this bacterium can be involved in the transmission of antimicrobial resistances, if it is released with manure as organic fertilizer on agricultural fields. The genetic content of the strain was compared to that of two other A. bohemicus strains, the type strain ANC 3994T, isolated from forest soil, and KCTC 42081, originally described as A. pakistanensis, a metal resistant strain isolated from a wastewater treatment pond. In contrast to the forest soil strain, both strains from anthropogenically impacted sources showed genetic features indicating their evolutionary adaptation to the anthropogenically impacted environments. Strain QAC-21b will be used as model strain to study the transmission of antimicrobial resistance to environmentally adapted Acinetobacter in agricultural environments receiving high content of pollutants with organic fertilizers from livestock husbandry. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a major causative agent of nosocomial infections and its outer membrane vesicles (AbOMVs) have been shown to be involved in pathogenicity by transporting virulence factors and transferring information for communication between pathogens and host cells. Despite the fact that the infected sites of A. baumannii such as lungs and skin soft tissues are hypoxic, most studies on AbOMV virulence have used AbOMVs prepared under aerobic conditions. The present study aims to elucidate the protein profile and pathogenic impact of AbOMVs released under hypoxic condition. AbOMVs were isolated from A. baumannii under normoxic and hypoxic conditions, and their protein profiles were compared. The different effects of both normoxic and hypoxic AbOMVs in cytokine response from mouse macrophages, cytotoxicity to the human lung epithelial cells, and bacterial invasion were then investigated. Our results showed that A. baumannii under hypoxia released larger amounts of OMVs with different protein profiles. Although the cytotoxic effect of AbOMVs from normoxia and hypoxia were comparable, AbOMVs from normoxia induced higher TNF-α production and invasion of Staphylococcus aureus and Pseudomonas aeruginosa than those from hypoxia. On the other hand, AbOMVs significantly enhanced A. baumannii invasion into lung epithelial cells in a dose-dependent manner. These results clearly demonstrate that AbOMVs released from normoxic and hypoxic have different impacts in pathogenesis. This finding provides new insight into the complex interactions between A. baumannii, coinfecting pathogens and host cells via OMVs, in particular the different pathogenic effects of AbOMVs under normoxic and hypoxic conditions. [ABSTRACT FROM AUTHOR]

Biofilm production facilitates microbial colonization of wounds and catheters. Acinetobacter baumannii produces high levels of biofilm and causes difficult-to-treat nosocomial infections. Candida albicans is another strong biofilm producer which may facilitate A. baumannii adhesion by providing hyphae-mediated OmpA-binding sites. Here we tested the potential of 2′-hydroxychalcones to inhibit dual-species biofilm production of A. baumannii and Candida spp., and further predicted the mechanism of structure-related difference in activity. The results suggest that 2′-hydroxychalcones exhibit potent activity against Candida spp./A. baumannii dual-species biofilm production. Particularly active was trifluoromethyl-substituted derivative (p-CF3), which decreased C. albicans/A. baumannii biomass produced on vein-indwelling parts of the central venous catheterization set by up to 99%. Further, higher OmpA-binding affinity was also calculated for p-CF3, which together with demonstrated significant ompA-downregulating activity, suggests that superior antibiofilm activity of this chalcone against the tested dual-species community of A. baumannii is mediated through the OmpA. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a nosocomial pathogen that can be resistant to antibiotics by rapidly modulating its anti-drug mechanisms. The multidrug-resistant A. baumannii has been considered one of the most threatening pathogens to our society. Biofilm formation and persistent cells within the biofilm matrix are recognized as intractable problems, especially in hospital-acquired infections. Poly-β-1,6-N-acetyl-glucosamine (PNAG) is one of the important building blocks in A. baumannii’s biofilm. Here, we discover a protein phosphoryl-regulation on PNAG deacetylase, AbPgaB1, in which residue Ser411 was phosphorylated. The phosphoryl-regulation on AbPgaB1 modulates the product turnover rate in which deacetylated PNAG is produced and reflected in biofilm production. We further uncovered the PgaB deficient A. baumannii strain shows the lowest level of biofilm production but has a high minimal inhibition concentration to antibiotic colistin and tetracycline. Based on bactericidal post-antibiotic effects and time-dependent killing assays with antibacterial drugs, we claim that the PgaB-deficient A. baumannii converts to colistin-tolerant cells. This study utilizes a biofilm-independent colistin-tolerant model of A. baumannii to further investigate its characteristics and mechanisms to better understand clinical outcomes. [ABSTRACT FROM AUTHOR]

Acinetobacter baumannii is a strictly aerobic, nonmotile, nonfermenting, gram-negative bacillus. It is a highly infectious and invasive pathogen with high mortality and morbidity rates among immunodeficient patients. Due to increasing levels of drug resistance and the inefficiency of existing antimicrobial treatments, it is crucial to develop novel agents to control this pathogen. Several recent studies have investigated virulence factors that are associated with the pathogenesis of A. baumannii , and could thus serve as novel therapeutic targets. The present review comprehensively summarizes the current understanding of these virulence factors and their mechanisms in A. baumannii. We also highlight factors that could be potential therapeutic targets, as well as list candidate virulence factors for future researchers and clinical practitioners. [ABSTRACT FROM AUTHOR]