Antibiotic Resistance Mechanisms Research Articles

Cross-talk of MLST and transcriptome unveiling antibiotic resistance mechanism of carbapenem resistance Acinetobacter baumannii clinical strains isolated in Guiyang, China.

Acinetobacter baumannii (A. baumannii) is an important opportunistic pathogen causing nosocomial infection in the clinic. The occurrence rate of antibiotic resistance is increasing year by year, resulting in a highly serious situation of bacterial resistance. To better understand the local epidemiology of multidrug-resistant A. baumannii, an investigation was conducted on the antibiotic resistance of different types of A. baumannii and its relationship with the genes of A. baumannii. Furthermore, the molecular mechanism underlying antibiotic resistance in A. baumannii was investigated through transcriptome analysis. These results showed that a total of 9 STs were detected. It was found that 99% of the strains isolated in the hospital belonged to the same STs, and the clone complex CC208 was widely distributed in various departments and all kinds of samples. Furthermore, these A. baumannii strains showed high resistance to ertapenem, biapenem, meropenem, and imipenem, among which the resistance to ertapenem was the strongest. The detection rate of bla OXA-51 gene in these carbapenem resistance A. baumannii (CRAB) reached 100%; Additionally, the transcriptome results showed that the resistance genes were up-regulated in resistance strains, and these genes involved in biofilm formation, efflux pumps, peptidoglycan biosynthesis, and chaperonin synthesis. These results suggest that the CC208 STs were the main clonal complex, and showed high carbapenem antibiotic resistance. All these resistant strains were distributed in various departments, but most of them were distributed in intensive care units (ICU). The bla OXA-23 was the main antibiotic resistance genotype; In summary, the epidemic trend of clinical A. baumannii in Guiyang, China was analyzed from the molecular level, and the resistance mechanism of A. baumannii to carbapenem antibiotics was analyzed with transcriptome, which provided a theoretical basis for better control of A. baumannii.

Detection of antibiotic resistance mechanism and research on new anti-infection measures based on big data analysis

Abstract In this paper, a prototype network model is used for event extraction of antibiotic resistance and new anti-infection information, and also limited or small amount of labeled data is used to obtain a prediction model with excellent generalization performance. The Bi-LSTM framework for antibiotic resistance element detection is a mainstream sequence annotation framework that combines bidirectional long- and short-term memory networks, and its main idea is to obtain contextual information through bidirectional long- and short-term memory networks, and to fuse antibiotic as well as bacterial entity information, and trigger word information for the assignment of element roles. In the keyword co-occurrence analysis, combination therapy and phage had the highest frequency with a centrality of 0.57 and 0.59, respectively, indicating that phage-antibiotic therapy effectively treated patients with drug-resistant bacterial infections. There was a high correlation (r=0.57) between the number of days of ICU stay and the number of days of phage-antibiotic combination therapy, which was further analyzed to show that the number of days of fever, the number of days of indwelling urinary catheter, and the number of days of mechanical ventilation, phage-antibiotic combination days are more important influencing factors.

Comparison of the antibiotic resistance mechanisms in a gram-positive and a gram-negative bacterium by gene networks analysis.

Nowadays, the emergence of some microbial species resistant to antibiotics, both gram-positive and gram-negative bacteria, is due to changes in molecular activities, biological processes and their cellular structure in order to survive. The aim of the gene network analysis for the drug-resistant Enterococcus faecium as gram-positive and Salmonella Typhimurium as gram-negative bacteria was to gain insights into the important interactions between hub genes involved in key molecular pathways associated with cellular adaptations and the comparison of survival mechanisms of these two bacteria exposed to ciprofloxacin. To identify the gene clusters and hub genes, the gene networks in drug-resistant E. faecium and S. Typhimurium were analyzed using Cytoscape. Subsequently, the putative regulatory elements were found by examining the promoter regions of the hub genes and their gene ontology (GO) was determined. In addition, the interaction between milRNAs and up-regulated genes was predicted. RcsC and D920_01853 have been identified as the most important of the hub genes in S. Typhimurium and E. faecium, respectively. The enrichment analysis of hub genes revealed the importance of efflux pumps, and different enzymatic and binding activities in both bacteria. However, E. faecium specifically increases phospholipid biosynthesis and isopentenyl diphosphate biosynthesis, whereas S. Typhimurium focuses on phosphorelay signal transduction, transcriptional regulation, and protein autophosphorylation. The similarities in the GO findings of the promoters suggest common pathways for survival and basic physiological functions of both bacteria, including peptidoglycan production, glucose transport and cellular homeostasis. The genes with the most interactions with milRNAs include dpiB, rcsC and kdpD in S. Typhimurium and EFAU004_01228, EFAU004_02016 and EFAU004_00870 in E. faecium, respectively. The results showed that gram-positive and gram-negative bacteria have different mechanisms to survive under antibiotic stress. By deciphering their intricate adaptations, we can develop more effective therapeutic approaches and combat the challenges posed by multidrug-resistant bacteria.

Genomic Characteristics of a Carbapenem-Resistant Klebsiella pneumoniae Co-Carrying bla NDM-5 and bla KPC-2 Capsular Type KL25 Recovered from a County Level Hospital in China.

Hypervirulent carbapenem-resistant K. pneumoniae (hv-CRKP) has been spreading rapidly worldwide. Here, we investigated the genomic characteristics of ST11 K. pneumoniae isolate SM117 with capsular serotype KL25, co-carrying bla NDM-5, two copies of bla KPC-2 and multiple plasmid-borne virulence genes from a county level hospital in China. Antimicrobial susceptibility of K. pneumoniae SM117 was evaluated. The Illumina NovaSeq 6000 and Oxford Nanopore MinION platforms were applied to sequence the genome and then de novo assembled. The genome sequence was annotated using the NCBI Prokaryotic Genome Annotation Pipeline and further subjected to identify the sequence type (ST), capsular type, antibiotic resistance genes, plasmid replicon types and virulence genes. The phylogenetic analysis was performed based on the core genome single nucleotide polymorphisms (cgSNPs) using CSI Phylogeny 1.4, and further visualized by Interactive Tree of Life (iTOL) V5 web server. The whole-genome sequence of K. pneumoniae SM117 is made up of eight contigs totaling 6,104,486 bp, contain a 5,612,620 bp single chromosome and seven plasmids. The isolate was assigned to ST11 with capsular serotype KL25, co-carrying including bla NDM-5, bla KPC-2 and multiple plasmid-borne virulence genes including rmpA2 and aerobactin genes iucABCD-iutA. The coexistence of bla KPC and bla NDM in K. pneumoniae strains exhibit a high degree of resistance to β-lactam antibiotics. The strain SM117 also carries multiple antibiotic resistance genes, making it resistant to all antibiotics except polymyxin. The closest relative of K. pneumoniae C793 was identified in 2023 from a hospital surface sample in Zhejiang, China, with just 52 SNPs difference. This study reported the genomic characteristics of a multidrug-resistant ST11 K. pneumoniae with capsular serotype KL25, co-carrying bla NDM-5, two copies of bla KPC-2 genes and multiple plasmid-borne virulence genes in China. These findings will provide important knowledge of the antibiotic resistance mechanisms, genomic epidemiological characteristics and transmission dynamics of multidrug-resistant K. pneumoniae.

In Vitro Effects of Silver Nanoparticles on Pathogenic Bacteria and on Metabolic Activity and Viability of Human Mesenchymal Stem Cells.

The rapid development of nanotechnology has led to the use of silver nanoparticles (Ag-NPs) in various biomedical fields. However, the effect of Ag-NPs on human mesenchymal stem cells (hMSCs) is not fully understood. Moreover, too frequent an exposure to products containing nanosilver in sublethal amounts raises widespread concerns that it will lead to the development of silver-resistant microorganisms. Therefore, this study aimed to evaluate the mechanism of action of Ag-NPs on hMSCs by analyzing the cellular uptake of Ag-NPs by the cells and its effect on their viability and to assess antimicrobial activity of Ag-NPs against emerging bacterial strains, including multidrug-resistant pathogens. For metabolic activity and viability evaluation, hMSCs were incubated with different concentrations of Ag-NPs (14 μg/mL, 7 μg/mL, and 3.5 μg/mL) for 10 min., 1 h and 24 h and subsequently analyzed for their viability by live-dead staining and metabolic activity by the MTS assay. The effect of Ag-NPs on bacterial pathogens was studied by determining their minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). In conclusion, it was observed that exposure of hMSCs to Ag-NPs of size <10 nm has no cytotoxic effect on the metabolic activity of the cells at the concentration of 3.5 μg/mL, with minimal cytotoxic effect being observed at the concentration of 14 μg/mL after 24 h of incubation. Our findings also confirmed that Ag-NPs at the concentration of 4 μg/mL are effective broad-spectrum bactericidal agents, regardless of the antibiotic-resistance mechanism present in bacteria.

THE PERSPECTIVE OF N-CHLOROTAURINE INFLUENCE ON MULTI-DRUF RESISTANT MICROORGANISMS ASSOCIATED WITH DENTAL IMPLANTATION FAILURE

Introduction: Microorganisms colonizing the surface of dental implants can form biofilms that protect their microcolonies from mechanical impact, antimicrobial compounds, and immune factors. Prophylactic use of antibiotics during dental implantation can exert selective pressure on the development of antimicrobial resistance. In this context, N-chlorotaurine (NCT) is a promising compound to combat resistant agents.&#x0D; The aim is to investigate the efficacy of NCT against pathogens associated with dental implant complications.&#x0D; Materials and methods. A buffered 1% NCT solution adjusted to pH 7.1 was used for the study. Multidrug-resistant cultures (MDR) were identified. Determination of NCT activity was carried out by the agar and suspension method.&#x0D; The results. After standard antibiotic susceptibility testing, cultures classified as MDR were selected. 12 cultures were included in the MDR group, including seven S. aureus, three K. pneumoniae and three A. baumannii. MDR cultures of S. aureus had MRSA profile. Production of carbapenemases was confirmed in all MDR K. pneumoniae isolates, 2 of them showed production of metallo-β-lactamases. MDR isolates A. baumannii were also producers of metallo-β-lactamases. All 12 investigated MDR cultures were sensitive to NCT, the diameter of the zones of growth retardation was at least 15 mm. No significant difference was found between the effectiveness of 1% NCT solution against microorganisms with different mechanisms of drug resistance (t-test, p&gt;0.05). When conducting studies in a liquid environment, a 1% NCT solution showed similar results, all MDR cultures were sensitive and showed a decrease in the microbial load (CFU/ml) by at least 2 log10 steps already 15 minutes after the start of the study. Notably, there was no significant difference in the efficacy of NCT against conditionally susceptible and MDR agents.&#x0D; Conclusions. Reducing the threat and burden of MDR pathogens is one of the goals announced by WHO and the Centres for Disease Control. Our study clearly confirms that various antibiotic resistance mechanisms do not affect the bactericidal activity of NCT, making it promising for the treatment of resistant infections.

Effect of Different Culture Media on Pseudomonas aeruginosa Biofilm Formation

The opportunistic pathogen Pseudomonas aeruginosa (PA) causes nosocomial infections, and it is the most common pathogen that can form biofilm. PA biofilm formation is important as an environmental bacterium in hospital wastewater, in vivo, in the environment, and in infection control. Besides many antibiotic resistance mechanisms, biofilms may play an important role as in PA forming biofilms that have a minimum inhibitory concentration (MIC) for antibiotics up to 1,000-fold higher than that of planktonic bacteria. Multiple biofilm-specific mechanisms contribute to the high levels of antibiotic resistance. Therefore, PA biofilm-associated infections lead to important clinical outcomes. The aim was to investigate the efficacy of four different culture media used in two biofilm formation protocols on the assessment of biofilm production by 11 PA isolated from hospital wastewater. The crystal violet microtiter plate-based method was used to evaluate the quantification of the biofilm formation capacity of PA. Results of culture media used in the formation of biofilm capacity were; TSB with %1 glucose 0.0 %, 63.6%, and 36.4%; BHI 18.2%, 36.4%, and 45.5%; LBB 9.1%, 27.3%, and 63.6% of isolates were strong, moderate, and weak biofilm producers, respectively. However, in MHB, 27.3%, 63.6%, and 9.1% of isolates were moderate, weak, and non-biofilm producers, respectively. The biofilm levels in protocol one were higher than the other protocol used (OD570). PA biofilm formation and quantification in these media used may help to search for antibiofilm agents in laboratories to prevent the spread of antimicrobial resistance, develop effective precautions, and prevent PA infections in hospitals.

Microfluidic Giant Polymer Vesicles Equipped with Biopores for High-Throughput Screening of Bacteria.

Understanding the mechanisms of antibiotic resistance is critical for the development of new therapeutics. Traditional methods for testing bacteria are often limited in their efficiency and reusability. Single bacterial cells can be studied at high throughput using double emulsions, although the lack of control over the oil shell permeability and limited access to the droplet interior present serious drawbacks. Here, a straightforward strategy for studying bacteria-encapsulating double emulsion-templated giant unilamellar vesicles (GUVs) is introduced. This microfluidic approach serves to simultaneously load bacteria inside synthetic GUVs and to permeabilize their membrane with the pore-forming peptide melittin. This enables antibiotic delivery or the influx of fresh medium into the GUV lumen for highly parallel cultivation and antimicrobial efficacy testing. Polymer-based GUVs proved to be efficient culture and analysis microvessels, as microfluidics allow easy selection and encapsulation of bacteria and rapid modification of culture conditions for antibiotic development. Further, a method for in situ profiling of biofilms within GUVs for high-throughput screening is demonstrated. Conceivably, synthetic GUVs equipped with biopores can serve as a foundation for the high-throughput screening of bacterial colony interactions during biofilm formation and for investigating the effect of antibiotics on biofilms.

Molecular characterization of the multi-drug resistant Myroides odoratimimus isolates: a whole genome sequence-based study to confirm carbapenem resistance.

The bacteria belonging to the Myroides genus are opportunistic pathogens causing community or hospital-acquired infections that result in treatment failure due to antibiotic resistance. This study aimed to investigate molecular mechanisms of antibiotic resistance, clonal relatedness, and the biofilm forming capacity of the 51 multi-drug resistant Myroides odoratimimus. All isolates were screened for blaKPC, blaOXA, blaVIM, blaIMP, blaMUS, blaTUS, blaNDM, and blaB genes by using PCR amplification. Whole genome sequencing (WGS) was applied on three randomly selected isolates for further investigation of antibiotic resistance mechanisms. Clonal relatedness was analyzed by Pulsed-field gel electrophoresis (PFGE) and the microtiter plate method was used to demonstrate biofilm formation. All isolates were positive for biofilm formation. PCR analysis resulted in a positive for only the blaMUS-1 gene. WGS identified blaMUS-1, erm(F), ere(D), tet(X), and sul2 genes in all strains tested. Moreover, the genomic analyses of three strains revealed that genomes contained a large number of virulence factors (VFs). PFGE yielded a clustering rate of 96%. High clonal relatedness, biofilm formation, and multi-drug resistance properties may lead to the predominance of these opportunistic pathogens in hospital environments and make them cause nosocomial infections.

Antibiotic adjuvants: synergistic tool to combat multi-drug resistant pathogens.

The rise of multi-drug resistant (MDR) pathogens poses a significant challenge to the field of infectious disease treatment. To overcome this problem, novel strategies are being explored to enhance the effectiveness of antibiotics. Antibiotic adjuvants have emerged as a promising approach to combat MDR pathogens by acting synergistically with antibiotics. This review focuses on the role of antibiotic adjuvants as a synergistic tool in the fight against MDR pathogens. Adjuvants refer to compounds or agents that enhance the activity of antibiotics, either by potentiating their effects or by targeting the mechanisms of antibiotic resistance. The utilization of antibiotic adjuvants offers several advantages. Firstly, they can restore the effectiveness of existing antibiotics against resistant strains. Adjuvants can inhibit the mechanisms that confer resistance, making the pathogens susceptible to the action of antibiotics. Secondly, adjuvants can enhance the activity of antibiotics by improving their penetration into bacterial cells, increasing their stability, or inhibiting efflux pumps that expel antibiotics from bacterial cells. Various types of antibiotic adjuvants have been investigated, including efflux pump inhibitors, resistance-modifying agents, and compounds that disrupt bacterial biofilms. These adjuvants can act synergistically with antibiotics, resulting in increased antibacterial activity and overcoming resistance mechanisms. In conclusion, antibiotic adjuvants have the potential to revolutionize the treatment of MDR pathogens. By enhancing the efficacy of antibiotics, adjuvants offer a promising strategy to combat the growing threat of antibiotic resistance. Further research and development in this field are crucial to harness the full potential of antibiotic adjuvants and bring them closer to clinical application.

Staphylococcus aureus functional amyloids catalyze degradation of β-lactam antibiotics

Antibiotic resistance of bacteria is considered one of the most alarming developments in modern medicine. While varied pathways for bacteria acquiring antibiotic resistance have been identified, there still are open questions concerning the mechanisms underlying resistance. Here, we show that alpha phenol-soluble modulins (PSMαs), functional bacterial amyloids secreted by Staphylococcus aureus, catalyze hydrolysis of β-lactams, a prominent class of antibiotic compounds. Specifically, we show that PSMα2 and, particularly, PSMα3 catalyze hydrolysis of the amide-like bond of the four membered β-lactam ring of nitrocefin, an antibiotic β-lactam surrogate. Examination of the catalytic activities of several PSMα3 variants allowed mapping of the active sites on the amyloid fibrils’ surface, specifically underscoring the key roles of the cross-α fibril organization, and the combined electrostatic and nucleophilic functions of the lysine arrays. Molecular dynamics simulations further illuminate the structural features of β-lactam association upon the fibril surface. Complementary experimental data underscore the generality of the functional amyloid-mediated catalytic phenomenon, demonstrating hydrolysis of clinically employed β-lactams by PSMα3 fibrils, and illustrating antibiotic degradation in actual S. aureus biofilms and live bacteria environments. Overall, this study unveils functional amyloids as catalytic agents inducing degradation of β-lactam antibiotics, underlying possible antibiotic resistance mechanisms associated with bacterial biofilms.

A brief insight into Citrobacter species - a growing threat to public health

Citrobacter spp. are Gram-negative, non-spore forming, rod-shaped, facultative anaerobic bacteria from the Enterobacteriaceae family often found in soil, sewage, sludge, water, food, and the intestinal tracts of animals and humans. Several members of Citrobacter spp. especially C. freundii, C. koseri, C. braakii are frequently detected in newborn illnesses, urinary tract infections, and patients with severe underlying conditions, including hypertension, diabetes, cancer, and respiratory infections, or those who are immunocompromised. Strains of Citrobacter spp. can spread vertically or horizontally from carriers or other hospital sources and thus cause nosocomial infections in hospital settings. A total of 19 Citrobacter genomospecies have been recognized based on genomics. It has been noted that the Citrobacter genus acquired antimicrobial resistance and virulence, including invasion, colonization, biofilm formation, and toxin production. The recent emergence and spread of antimicrobial resistance to β-lactams, carbapenems, fluoroquinolones, aminoglycosides, and colistin in Citrobacter spp. through chromosomal and plasmid-mediated resistance limits the empiric treatment options. Therefore, combination therapy involving costly and potentially hazardous antibiotics poses significant challenges in treating Citrobacter infections. Here we summarized the nomenclature of Citrobacter spp., clinical manifestations, epidemiology, pathogenesis, antibiotic resistance mechanisms, and treatments from various clinical samples. This review will expand our knowledge of the genomics and epidemiology of Citrobacter spp., enabling improved control of infections and the spread of these organisms.

Novel Approaches for Combating Antibiotic Resistance in Pathogenic Bacteria

Antibiotic resistance has historically been mostly linked to the established mechanisms such target alteration, efflux pumps, and enzymatic inactivation. Also, the ability of bacteria to form biofilm is a formidable challenge to antibiotic efficacy. The extracellular matrix forms a protective barrier that renders most antibiotics ineffective, by limiting the interaction of the drug to its target. The emergence of novel antibiotic resistance mechanisms in pathogenic bacteria and its rapid propagation, by means of horizontal gene transfer or mobile genetic elements, is a constant challenge to the medical community. Bacteria are able to exchange genetic material, conferring to antibiotic resistance, at a startling rate that helps them adapt quickly. This is especially dangerous when antibiotics use is abused. The field of antibiotic resistance has experienced a notable shift in the past few years, owing to the identification of new resistance mechanisms utilized by pathogenic bacteria to impede the effectiveness of antibiotics. Over the past decade researchers have been rigorously studying these resistance mechanisms and working on strategies to overcome the ordeal. Notably, nanoparticles research targeting antibiofilm therapy to combat the antibiotic-resistance mechanisms has been truly innovative. There have also been positive reports on utilizing the CRISPR-Cas as a promising next-generation antibiotic that can be used to eliminate antibiotic resistant genes (ARGs) and resensitize bacterial populations to antibiotics. This review aims for a comprehensive understanding of the ongoing research on antibiotic resistance mechanisms and strategies to counter them, the remaining challenges for widespread clinical implementation and the importance of continued research.

Prevalence of biofilm producing Acinetobacter baumannii clinical isolates: A systematic review and meta-analysis.

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. 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. 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%). 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.

Biological characteristics of the enteroaggregative &lt;i&gt;escherichia coli &lt;/i&gt;ont:h30 18-726 (no. B-8857) strain isolated from a patient with ulcerative colitis and a new method of pcr identification

The aim of the work is to assess biological properties of the enteroaggregativeEscherichia coli(EAgEC) strain isolated from the colon mucosa biopsy material from a patient with ulcerative colitis, and to develop a new method for PCR identification ofE. colistrains.&#x0D; Materials and methods.The study involved 46 patients with a verified ulcerative colitis. Isolation of 87E. colistrains was carried out by using colon biopsy material obtained during a standard endoscopic procedure. The description of the biochemical bacterial properties was combined with the molecular genetic detection of virulence determinants and presence of antibiotic resistance mechanisms. Using the “AmpliSense®Escherichiose — FL” reagent kit, a screening for the presence of diarrheagenicE. colistrains was performed, which revealed the presence of a single strain of EAgEC. The methodological approach to creating a new method for strain identification was based on the search for a unique genetic sequence within the glutamate decarboxylase(gad)gene.&#x0D; Results.E. coli18-726 can be described biochemically typical to its species. The studied strain was characterized by a multiple resistance phenotype (MDR) to antibiotics, as well as a lack of sensitivity to five bacteriophages. TheE. coli18-726 strain had a unique sequence of the gene encoding the O-antigen, which differed from 188 known O-antigens (ONT) and, by the identity of the nucleotide sequence of the gene encoding the synthesis of the H-antigen, the strain belonged to the H30 serovar. Thus, the antigenic formula of strain EAgEC 18-726 was expressed as ONT:H30. TheE. coli18-726 strain contained a significant spectrum of genes that enable properties of virulence(iss, capU, aggA, aggB, aggC, aggD, aap, aar)and resistance to antibiotics(blaCTX-M-15, blaTEM-1B, aadA1, aadA5, mph(A), catB3). A new method for identifying EAgEC in chronic bowel disease is based on the polymerase chain reaction method using primers for a specific region of bacterial glutamate decarboxylase(gad)gene.Conclusion.1) The biochemical, antigenic and molecular genetic properties ofE. coliONT:H30 18-726 strain (No. B-8857) isolated from a patient with histomorphologically diagnosed ulcerative colitis were analyzed. 2) A method for PCR identification of EAgEC strains based on the detection of a specific region within the bacterial genomic glutamate decarboxylase gene has been created and patented.

The Prevalence of Alert Pathogens and Microbial Resistance Mechanisms: A Three-Year Retrospective Study in a General Hospital in Poland.

The development of antibiotic resistance mechanisms hinders the treatment process. So far, there is limited data on the problem of bacterial resistance in hospitals in Central and Eastern Europe. Therefore, this study aimed to assess the prevalence of resistance mechanisms and alert pathogens based on reports regarding cultures of samples collected from general hospital patients in Poland in the period 2019-2021. This study examined the prevalence of resistance mechanisms and alert pathogens and the structure of microorganisms, including the type of diagnostic material in the hospital department. The frequency of occurrence and the trends were analysed based on Cochran's Q-test, relative change and the average annual rate of change (AARC). Of all 14,471 cultures, 3875 were positive for bacteria, and 737 were characterised by resistance mechanisms (19.0%). Alert pathogens were identified in 983 cases (24.6%), including pathogens isolated from blood samples. The most commonlyisolated bacteria were Escherichia coli (>20% of positive cultures), Enterococcus faecalis (7%), and Klebsiella pneumoniae (6%) increasing over time; Staphylococcus aureus (13%) was also found, but its proportion was decreasing over time. Extended-spectrum β-lactamase (ESBL) was the most frequent resistance mechanism in Internal Medicine (IM) (p < 0.001) and the Intensive Care Unit (ICU) (p < 0.01), as well as in ICU-COVID; this increased over the study period (AARC ↑34.9%). Methicillin-resistant Staphylococcus aureus (MRSA) (AARC ↓50.82%) and AmpC beta-lactamase (AARC ↓24.77%) prevalence fell over time. Also, the number of alert pathogens was dominant in the IM (p < 0.01) and ICU (p < 0.001). The most common alert pathogen was ESBL-positive E. coli. Gram-negative rods constitute a significant epidemiological problem in hospitals, especially the growing trend of ESBL in IM and ICU, which calls for increased control of sanitary procedures.

2780. Factors associated with Staphylococcus aureus susceptibility to two lytic bacteriophages; Comparison of phage host susceptibility using a reference set of S. aureus

Abstract Background Timely access is a major concern when considering phage therapy. Using susceptible phage is critical for the outcomes of phage therapy. For timely access to susceptible phages, identifying factors associated with phage susceptibility and building a reference set (RS) to compare host ranges is needed. Methods Phage K, which was a well-documented lytic S. aureus phage, and phage PBSA02, which was a new S. aureus phage from the Korean phage biobank, were used. The efficiency of plating (EOP), the relative activity of phage against specific strain/host strain, and growth kinetics assays were used to determine phage susceptibility. The isolates with high lytic activity (HLA) were defined as susceptible (EOP &amp;gt; 1%). The phage PBSA02 and phage K susceptible groups were compared to the non-susceptible groups to determine the associating factors of phage susceptibility. Results We constructed a methicillin-resistant S. aureus (MRSA) RS using the Australian Group on Antimicrobial Resistance (AGAR) repository. We determined the predominant sequence types (ST) of MRSA in Australia in 2021; ST22 accounted for 90.1% of HA-MRSA and 5 STs for 67.9% of CA-MRSA (figure 1). Then, we randomly selected 39 AGAR MRSA isolates corresponding to each ST from each region in Australia (figure 1). Phage PBSA02 exhibited HLA against 56.4% of the 39 RS isolates, while phage K exhibited HLA against 30.8%. Phage PBSA02 showed HLA against 100% of ST93 and ST30 isolates following ST1 (62.5%), ST22 (37.5%), ST5 (20%) and ST45 (16.7%); phage K showed HLA against 100% of ST30 and 62.5% of ST22 isolates but against none of the ST93 isolates. Phage PBSA02 susceptibility correlated with the ST of MRSA isolates and ciprofloxacin susceptibility (table 1). Isolates from pneumonia/empyema tended to show phage PBSA02 susceptibility (table 1). Phage K susceptibility was also significantly associated with the ST (table 2). Conclusion Our study demonstrated that phage susceptibility was associated with the STs of MRSA, and the RS could be useful for a standardised comparison of the phage host range. Phage PBSA02 with broad susceptibility to the RS could be useful for phage therapy. Further work includes antibiotic synergy testing and resistance mechanism identification in these isolates. Disclosures All Authors: No reported disclosures

Metabolomics Method in Understanding and Sensitizing Carbapenem-Resistant Acinetobacter baumannii to Meropenem.

Carbapenem-resistant Acinetobacter baumannii (CRAB) strains are prevalent worldwide and represent a major threat to public health. However, treatment options for infections caused by CRAB are very limited as they are resistant to most of the commonly used antibiotics. Consequently, understanding the mechanisms underlying carbapenem resistance and restoring bacterial susceptibility to carbapenems hold immense importance. The present study used gas chromatography-mass spectrometry (GC-MS)-based metabolomics to investigate the metabolic mechanisms of antibiotic resistance in clinically isolated CRAB. Inactivation of the pyruvate cycle and purine metabolism is the most typical characteristic of CRAB. The CRAB exhibited a reduction in the activity of enzymes involved in the pyruvate cycle, proton motive force, and ATP levels. This decline in central carbon metabolism resulted in a decrease in the metabolic flux of the α-ketoglutarate-glutamate-glutamine pathway toward purine metabolism, ultimately leading to a decline in adenine nucleotide interconversion. Exogenous adenosine monophosphate (AMP) and adenosine triphosphate (ATP) enhance the killing efficacy of Meropenem against CRAB. The combination of ATP and Meropenem also has a synergistic effect on eliminating CRAB persisters and the biofilm, as well as protecting mice against peritonitis-sepsis. This study presents a novel therapeutic modality to treat infections caused by CRAB based on the metabolism reprogramming strategy.

Progress on mechanisms of antibiotic resistance in Clostridioides difficile.

Clostridioides difficile (CD) is one of the most common pathogens causing health-care-associated infectious diarrhea and is listed by the U.S. Centers for Disease Control and Prevention as an urgent antibiotic resistance (AR) threat. Many resistance genes can be transferred between different CD strains present in the clinical setting, community, and environment. The antimicrobial resistance (AMR) of CD continues to evolve with the emergence and acquisition of new drug resistance mechanisms. CD has developed diverse drug resistance mechanisms, such as drug alteration, modification of the target site, and extrusion of drugs via efflux pumps. Researches have provided comprehensive knowledge about resistance mechanisms of macrolides and quinolones in CD. However, the mechanisms of resistance for metronidazole, vancomycin, and other therapeutic antibiotics against Clostridioides difficile infection (CDI) are only beginning to be elucidated. Some previously unfound mechanisms, such as plasmid-mediated drug resistance in CD, may also play an important role. In this review, we summarize the research progress on drug resistance mechanisms of CD with antimicrobial drugs already used clinically, such as metronidazole, vancomycin, and fidaxomicin, thereby providing the references for the clinical treatment and prevention of CDI, as well as the development of new antibacterial drugs and detection kits for drug resistant bacteria.

Morphological Characterization of Calcium Carbonate Nanoparticles Derived from Blood Cockle Shells (Anadara granosa)

Blood cockle shells are a type of shellfish with distinctive bright red color that is commonly found in coastal areas of Asia, a rich source of protein, vitamins, and minerals, including calcium carbonate. Calcium carbonate (CaCO3) is a common mineral supplement that slowly paved its way into nano research to support bone health that also used in the treatment of bone infections due to its inherent properties; reduces inflammation, promotes bone healing, crucial role in the immune system, and slow degradability. The latest invention on CaCO3 from cockle shells, is an alternative mode of drug delivery to circumvent existing antibiotic resistance mechanisms and allow direct drug delivery to the target site, using locally sourced ingredients to reduce long-term high production costs. A top-down approach CaCO3 nanoparticles from blood cockle (Anadara granosa) shells were developed, known as calcium carbonate nanoparticles (CNPs). This research aims to characterize the morphology of CNPs from blood cockle shells via Transmission Electron Microscopy (TEM), Scanning Electron Microscopy (SEM), and Atomic Force Microscopy (AFM). A homogeneously spherical CNPs. with a mean diameter of 36.5 ± 2.3 nm was observed on TEM, so as FESEM. From AFM, the mean diameter for CNPs was 38.6 ± 3.6 nm The CNPs were spherical in shape, homogenously scattered, less agglomerate, porous with rough surfaces. The size and morphology of the CNPs from blood cockle shells were suitable for transport carrier in situ bone infection, according to the results.