Carbapenems Research Articles

Identification of a potential inhibitor for New Delhi metallo-β-lactamase 1 (NDM-1) from FDA approved chemical library- a drug repurposing approach to combat carbapenem resistance

Superbugs producing New Delhi metallo-β-lactamase 1 (NDM-1) enzyme is a growing crisis, that is adversely affecting the global health care system. NDM-1 empowers the bacteria to inactivate entire arsenal of β-lactam antibiotics including carbapenem (the last resort antibiotic) and remains ineffective to all the available β lactamase inhibitors used in the clinics. Limited therapeutic option available for rapidly disseminating NDM-1 producing bacteria makes it imperative to identify a potential inhibitor for NDM-1 enzyme. With drug repurposing approach, in this study, we used virtual screening of available Food and Drug Administration (FDA) approved chemical library (ZINC12 database) and captured ‘adapalene’ (FDA drug) as a potent inhibitor candidate for NDM-1 enzyme. Active site docking with NDM-1, showed adapalene with binding energy −9.21 kcal/mol and interacting with key amino acid residues (Asp124, His122, His189, His250, Cys208) in the active site of NDM-1. Further, molecular dynamic simulation of NDM-1 docked with the adapalene at 100 ns displayed a stable conformation dynamic, with relative RMSD and RMSF in the acceptable range. Subsequently, in vitro enzyme assays using recombinant NDM-1 protein demonstrated inhibition of NDM-1 by adapalene. Further, the combination of adapalene plus meropenem (carbapenem antibiotic) showed synergistic effect against the NDM-1 producing carbapenem (meropenem) resistant clinical isolates (Escherichia coli and Klebsiella pneumoniae). Overall, our data indicated that adapalene can be a potential inhibitor candidate for NDM-1 enzyme that can contribute to the development of a suitable adjuvant to save the activity of carbapenem antibiotic against infections caused by NDM-1 positive gram-negative bacteria. Communicated by Ramaswamy H. Sarma

Bacterial and Antibiogram Profile of Urinary Tract Infection Patients in Tertiary Hospital, Surabaya, Indonesia

Highlights: Most urinary tract infections are caused by Gram-negative bacteria with E. coli being the most common bacteria. Antibiotics with the highest susceptibility for Gram-negative bacteria were ertapenem, meropenem, amikacin, and imipenem. Antibiotics with the highest susceptibility for Gram-positive bacteria mainly were susceptible to chloramphenicol,streptomycin, vancomycin, rifampin, tigecycline, teicoplanin, and ampicillin. Abstract: Urinary tract infection (UTI) is one of the most common bacterial infections. Inappropriate antibiotic use for UTI treatment may lead to antibiotic resistance. This study aimed to provide an updated bacterial and antibiogram profile from urine specimens of patients diagnosed with UTI. This study was a retrospective study using urine culture and antibiotic susceptibility test results obtained from Clinical Microbiology Laboratory in a tertiary general hospital in Surabaya, Indonesia for a two-month period patients. Most aged more than and/or equal to 59 years, in both sexes. Gram-negative bacteria, particularly , was being the most between June to July 2019. There were 215 patients with significant urine culture results of 54.4% from Escherichia coli female bacteria were carbapenem antibiotics and amikacin, while teicoplanin and vancomycin were some antibiotics susceptible to gramcommon bacteria that caused UTI, followed by K. pneumoniae. Some antibiotics with the highest susceptibility to gram-negative positive bacteria. This study result indicated that there was an urge to conduct local antibiogram profile investigation due to the low susceptibilities shown in recent empirical therapy recommendations, such as trimethoprim sulfamethoxazole, fluoroquinolone, nitrofurantoin, and fosfomycin.

Enhancement of Acinetobacter baumannii biofilm growth by cephem antibiotics via enrichment of protein and extracellular DNA in the biofilm matrices.

AimsThe aims were to determine the effects of subinhibitory concentrations of eight cephem and carbapenem antibiotics on the biofilm formation of Acinetobacter baumannii cells and examine their effects on pre‐established biofilms.Methods and ResultsEffects of antibiotics on biofilm formation were assayed using microtitre plates with polystyrene peg‐lids. Cefmetazole, ceftriaxone, ceftazidime and cefpirome increased the biomass of pre‐established biofilms on pegs in the range of their sub‐minimum inhibitory concentrations (MICs), whereas none increased biofilm formation by planktonic cells. Carbapenems had a negative effect. The constituents of antibiotic‐induced biofilms were analysed. Ceftriaxone or ceftazidime treatment markedly increased the matrix constituent amounts in the biofilms (carbohydrate, 2.7‐fold; protein, 8.9–12.7‐fold; lipid, 3.3–3.6‐fold; DNA, 9.1–12.2‐fold; outer membrane vesicles, 2.7–3.8‐fold and viable cells, 6.8–10.1‐fold). The antibiotic‐enhanced biofilms had increased outer membrane protein A and were resistant to the anti‐biofilm effect of azithromycin.ConclusionsSome cephems increased the biomass of pre‐established biofilms in the ranges of their sub‐MICs. The antibiotic‐enhanced biofilms possessed more virulent characteristics than normal biofilms.Significance and Impact of the StudyIncomplete administration of certain cephems following biofilm‐related Ac. baumannii infections could adversely cause exacerbated and chronic clinical results.

Proteomics profiling of ertapenem challenged major porin deficient carbapenem-resistant Klebsiella pneumoniae

Carbapenem-resistant Klebsiella pneumoniae (CRKP) is an urgent threat to human health. Major outer membrane proteins (OMPs) porin mutation is one important resistance mechanism of CRKP, and may also affect the inhibition activity of β-lactam and β-lactamase inhibitor combinations. The ertapenem-resistant K. pneumoniae strain 2018B120 with major porin mutations was isolated from a clinical patient. Genomic and time-series proteomic analyses were conducted to retrieve the ertapenem-challenged response of 2018B120. The abundance changing of proteins from PTS systems, ABC transporters, the autoinducer 2 (AI-2) quorum sensing system, and antioxidant systems can be observed. Overexpression of alternative porins was also noticed to balance major porins' defection. These findings added a detailed regulation network in bacterial resistance mechanisms and gave new insights into bypass adaptation mechanisms the porin deficient bacteria adopted under carbapenem antibiotics pressure. SignificanceOuter membrane porins deficiency is an important mechanism of carbapenem resistance in K. pneumoniae. Comprehensive genomic and proteomic profiling of an ertapenem-resistant K. pneumoniae strain 2018B120 gives a detailed systematic regulation network in bacterial resistance mechanisms. Overexpression of alternative porins to balance major porins' defection was noticed, giving new insights into bypass adaptation mechanisms of porin deficient bacteria.

Detection and characterisation of Bordetella hinzii in line-related bacteraemia and respiratory tract infection in Australia

Bordetella hinzii has emerged as an unusual cause of infection in immunocompromised patients, previously linked to zoonotic transmission. Antimicrobial susceptibility and genetic diversity of B.hinzii are poorly understood. This study reports phenotypic and genomic characteristics of the first four Australian isolates of B.hinzii obtained from elderly immunocompromised patients. Bordetella hinzii isolates were identified by MALDI-TOF and whole genome sequencing (WGS). Antibiotic susceptibility testing was performed using disk diffusion or E-test. Genomes of B.hinzii were analysed in global context. A phylogenetic tree was constructed of all isolates using Roary and a maximum-likelihood tree was generated from the core-snp alignment. Bordetella hinzii minimum inhibitory concentrations (MICs) were largely uniform with high MICs to ampicillin, ceftriaxone and ciprofloxacin and low MICs to meropenem and piperacillin-tazobactam. Genomic analysis of isolate sequences divided strains analysed into two phylogenetically distinct groups, with one Australian B.hinzii isolate (AUS-4) assigned to Group 1, and the remaining isolates (AUS1-AUS3 and AUS-5) to Group 2. Single nucleotide polymorphism (SNP) analysis revealed two isolates, AUS-1 and AUS-2, were closely related with 14 SNP differences between them. All other Australian isolates were unrelated to each and all other isolates from the international dataset. Bordetella hinzii appears to pose a risk to immunocompromised individuals but remains susceptible to extended spectrum β-lactam and carbapenem antibiotics. Genomic analysis suggested a dissemination of genetically distinct strains.

Pathogen Distribution and Antimicrobial Resistance of Early Onset Sepsis in Very Premature Infants: A Real-World Study.

Early onset sepsis (EOS) remains a potentially fatal newborn condition, especially in very preterm infants. Data on the pathogen distribution and antibiotic susceptibility patterns of EOS among very preterm infants are scarce but essential for the choice of empirical antibiotic administration. We sought to assess the epidemiologic characteristics and antibiotic susceptibility patterns of pathogens causing EOS among a cohort of very preterm infants in China. This prospective, observational study included a cohort of infants born at a gestational age (GA) less than 32weeks of 32 newborn intensive care units (NICUs) in China between January1, 2018 and December31, 2020. EOS was defined by isolation of pathogenic species from blood culture within 72h of birth. A total of 108 EOS cases (18.4 per 1000 admissions) were identified among 5865 very preterm infants. Incidence of EOS increased with the decrease of GA and birthweight. Escherichia coli (n = 44, 40.7%) was the most common pathogen, followed by Klebsiella spp. (n = 10, 9.3%). The distribution and proportion of pathogenic bacteria varied significantly by GA. E.coli and Klebsiella spp. showed high resistance to ampicillin and third-generation cephalosporins, while they showed good susceptibility to carbapenem antibiotics and piperacillin-tazobactam. Our data demonstrated that pathogens causing neonatal EOS showed high rates of resistance to ampicillin and third-generation cephalosporins. This raised questions about the best empirical antibiotic choice for preterm infants suspected of having EOS in low- and middle-income countries (LMICs).

In Vitro Efficacy of Biocompatible Zinc Ion Chelating Molecules as Metallo-β-Lactamase Inhibitor among NDM Producing Escherichia coli.

Objective This article assesses the effectiveness of captopril, tetracycline, and ciprofloxacin as metallo-β-lactamase (MBL) inhibitors against New Delhi metallo-β-lactamase (NDM)-producing Escherichia coli. Materials and Methods Twenty-four well-characterized carbapenem-resistant E. coli isolates which produced NDM ( n = 21) and Oxa-48-like enzymes ( n = 3) were used to assess the inhibitors. The positive control organism was designed by cloning the NDM gene into pET-24a plasmid and transforming it into expression vector E. coli BL21. All the proposed inhibitors were assessed for their interaction with MBLs using checkerboard minimum inhibitory concentration (MIC) assay with imipenem and meropenem. The fractional inhibitory concentration (FIC) index was calculated to assess the activity of molecules. Results The E. coli BL21 (DE3) pET-24a- bla NDM showed carbapenem resistance upon isopropyl β-D-1-thiogalactopyranoside induction and had MIC of 32 µg/mL for both imipenem and meropenem. For the test isolates, ∑FIC values of imipenem and meropenem with ethylenediaminetetraacetic acid (EDTA) ranged from 0.039 to 0.266 and 0.023 to 0.156, respectively. At a 256 µg/mL concentration, captopril had ∑FIC index value for imipenem and meropenem as 0.133 to 0.375 and 0.133 to 0.188, respectively. The tetracycline and ciprofloxacin in combination with meropenem/imipenem showed indifferent results. Conclusion Among the three molecules tested, captopril had MBL inhibitory activity, but the concentration required for inhibition was beyond the therapeutic safety levels. Ciprofloxacin and tetracycline had weak or no MBL inhibitory activity. Checkerboard MIC of EDTA with carbapenem antibiotic and control organism with NDM enzyme production helped us create a reference system for comparing and assessing the results of potential MBL inhibitors in future.

Microbial Resistance to Carbapenems in Effluents from Gynaecological, Paediatric and Surgical Hospital Units.

The aim of this work is to identify and count antimicrobial resistance (AMR) in hospital effluents (HEs) of 2 units of the University Hospital Mohamed VI the Mother and Child Hospital (MCH) and the Ar-razi Surgical Hospital (ArzH), and to compare the two hospital units in terms of ARMs and seasonal variation. Each HE was sampled during 2016 and 2017. After identification of the pathogenic strains and determination of AMR, the results were reported for 24 ABs, including 3 carbapenems (CBP), and their consumption rates. The Predicted environmental concentration (PEC) rate of carbapenems in the HE of the study sites is calculated. A comparative analysis of the AMR of the isolated bacterial species was performed and related to the evolution of PEC in HEs. In the ArzH effluents:15 strains isolated, 7 are carbanepenem-resistant Enterobacteria (CRE) and are resistant to at least one of the 3 carbapenems tested. ArzH and MCH effluents respectively show some similarities: 26.87% and 28.57% of isolated bacteria are resistant to ertapenem while 43.48% and 57.14% are resistant to meropenem. However, for imipenem, the MCH effluent has a higher percentage of bacterial antibiotic resistance than ArzH. In addition, the percentage of resistance in each hospital unit effluent is mainly in relation with the increasing antibiotic consumption and predicted environmental values PEC for very antibiotic in each unit in the same period.

Risk factors and resistance patterns of invasive Acinetobacter Baumannii infection in Children

Objective: To understand the risk factors and antibiotics-resistant patterns of invasive Acinetobacter baumannii infection in Children. Methods: This retrospective study was conducted in 6 tertiary hospitals from January 2016 to December 2018. The basic information, clinical data and the results of antimicrobial susceptibility testing were collected from the 98 pediatric inpatients with Acinetobacter baumannii isolated from blood or cerebrospinal fluid and analyzed. According to the susceptibility of the infected strains to carbapenems, they were divided into carbapenem-sensitive Acinetobacter baumannii (CSAB) group and carbapenem-resistant Acinetobacter baumannii (CRAB) group. According to the possible sources of infection, they were divided into nosocomial infection group and community infection group. Chi-square test or Fisher exact test were used to analyze categorical variables and rank sum test were used to analyze continuous variables. The risk factors of invasive CRAB infection in children were analyzed by Logistic regression. Result: There were 56 males and 42 females in 98 cases. The onset age of patients was 8 (2, 24) months. There were 62 cases (63%) from rural area. A total of 87 cases (89%) were confirmed with bloodstream infection, and 12 cases (12%) confirmed with meningitis (1 case was accompanied with bloodstream infection). In these patients, 66 cases (67%) received invasive medical procedures or surgery, 54 cases (55%) received carbapenems-containing therapy. Twenty-four cases were infected with CRAB, and 74 cases with CSAB. The onset age of cases in CRAB group was lower than that in CSAB group (4 (1, 9) vs. 10 (4, 24) months, Z=-2.16, P=0.031). The proportions of hospitalization in intensive care unit, carbapenem antibiotics using, pneumonia and adverse prognosis in CRAB group were higher than those in CSAB group (6 cases (25%) vs. 4 cases (5%), 18 cases (75%) vs. 36 cases (49%), 17 cases (71%) vs. 17 cases (23%), 6 cases (25%) vs. 4 cases (5%), χ2=5.61, 5.09, 18.32, 5.61, all P<0.05). Seventy-seven cases were nosocomial infection and 21 cases were hospital-acquired infection. The proportion of children hospitalized in high-risk wards for nosocomial infections, length of hospitalization, number of antimicrobial therapy received and duration of antimicrobial therapy were higher in the hospital associated infection group than those in the community acquired infection group (all P<0.05). Logistic regression analysis showed that children from rural area (OR=8.42, 95%CI 1.45-48.88), prior mechanical ventilation (OR=12.62, 95%CI 1.31-121.76), and prior antibiotic therapy (OR=4.90, 95%CI 1.35-17.72) were independent risk factors for CRAB infection. The resistance percentage of CSAB isolates to many classes of antibiotics was <6% except to gentamicin, which was as high as 20% (13/65). All CRAB isolates of resistant to ampicillin-sulbactam (20/20), cefepime (23/23), piperacillin (17/17), meropenem (23/23) and imipenem (24/24) were 100%. The resistance percentage to other antibiotics were up to 42%-96%. Conclusions: Most of invasive Acinetobacter baumannii infection in children in China are hospital-acquired. The outcome of invasive CRAB infection was poorer than that of CSAB infection. The drug resistance rate of CRAB strains isolated is high. Living in rural area, prior invasive mechanical ventilation and prior antibiotic therapy were independent risk factors for invasive CRAB infection. The prevention and control of nosocomial infection and appropriate use of antibiotics to reduce Acinetobacter baumannii infection.

Structure-Activity Relationship of Penem Antibiotic Side Chains Used against Mycobacteria Reveals Highly Active Compounds.

The rise of antibiotic-resistant Mycobacterium tuberculosis and non-tuberculous mycobacterial infections has placed ever-increasing importance on discovering new antibiotics to treat these diseases. Recently, a new penem, T405, was discovered to have strong antimicrobial activity against M. tuberculosis and Mycobacteroides abscessus. Here, a penem library of C2 side-chain variants was synthesized, and their antimicrobial activities were evaluated against M. tuberculosis H37Rv and M. abscessus ATCC 19977. Several new penems with antimicrobial activity stronger than the standard-of-care carbapenem antibiotics were identified with some candidates improving on the activity of the lead compound, T405. Moreover, many candidates showed little or no increase in the minimum inhibitory concentration in the presence of serum compared to the highly protein-bound T405. The penems with the strongest activity identified in this study were then biochemically characterized by reaction with the representative l,d-transpeptidase LdtMt2 and the representative penicillin-binding protein d,d-carboxypeptidase DacB2.

Predictive Value of a Quick Pitt Bacteremia Score for Prognosis of Patients with Bloodstream Infection Secondary to Urinary Tract Infection: A Retrospective Cohort Study.

PurposeTo analyze the effect of a quick Pitt bacteremia score (qpitt) on the prognosis of patients with bloodstream infection (BSI) secondary to urinary tract infection (UTI) and to further explore its use in aiding appropriate selection of initial antibiotic treatment.MethodsMedical records of patients with BSIs secondary to UTIs who were admitted to our hospital from January 2018 to December 2020 were retrospectively collected. To screen for independent risk factors, logistic analysis was conducted on statistically significant variables. The receiver operating characteristic (ROC) curve was drawn with prognosis and death as the state variables to evaluate the predictive value. Patients were grouped by qpitt 2-point cutoff, to explore the impact of initial antimicrobial treatment regimens on poor prognosis and death in different subgroups. Poor prognosis was defined as a hospital length of stay (HLOS) ≥14 days or death within 28 days from BSI onset (ie, 28-day death).ResultsA total of 266 patients were included in this study. In BSIs secondary to UTIs, we observed a pathogenic composition of 77.44% Gram-negative bacteria, 19.55% Gram-positive bacteria, and 3.01% fungi. The qpitt had poor predictive value for poor prognosis [area under ROC (AUROC) = 0.653, p < 0.001], while it had a high predictive value for death (AUROC = 0.890, p < 0.001). For patients with a qpitt ≥2, the poor prognosis and death rates of patients who were initially treated with carbapenem antibiotics were lower (p < 0.01). In comparison, initial treatment with carbapenem antibiotics had no significant effect on prognosis and death rates in patients with qpitt <2 (p > 0.1).ConclusionThe qpitt is highly predictive for death in patients with BSIs secondary to UTIs and can be used to inform first-line antibiotic treatment strategy.

Carbapenems versus Cephalosporin or Piperacillin-Tazobactam as Perioperative Antibiotic Prophylaxis in Liver Transplant Recipients with Model for End-Stage Liver Disease Scores of ≥30: A Retrospective Study in a Chinese Population.

ObjectivePerioperative prophylaxis, commonly with a third-generation cephalosporin plus ampicillin or piperacillin-tazobactam, is usually employed to prevent infections in liver transplantation (LT) recipients. Patients with a high Model for End-Stage Liver Disease (MELD) score have an increased infection risk after LT. However, whether carbapenems could be used as surgical prophylaxis in these high-risk patients remains unclear. Therefore, this study aimed at comparing the effectiveness of carbapenems with that of cephalosporin or piperacillin-tazobactam for surgical prophylaxis in high-risk LT recipients with a MELD score ≥30.Design or MethodsThis retrospective study included adult patients with a MELD score ≥30 who underwent LT between May 2018 and September 2020. We comparatively analyzed the infection rate and outcome between patients using cefoperazone-sulbactam or piperacillin-tazobactam and those using carbapenems as surgical prophylaxis.ResultsThis study included 105 LT recipients. Seventy-eight and 27 patients used non-carbapenem and carbapenem antibiotics, respectively, as surgical prophylaxis. The corresponding infection incidence rates within 30 days were 38.5% and 66.7% (p = 0.011). Multivariate analysis revealed that reoperation and the Child–Pugh score were independent risk factors for infections within 30 days after LT. The following four risk factors were associated with the 180-day post-LT survival: MELD score, vascular complication, intra-abdominal bleeding, and infection with carbapenem-resistant organisms (CROs). There was no significant difference in CRO infection incidence between the carbapenem and non-carbapenem groups (18.5% vs 11.5%; p = 0.345).ConclusionCarbapenem use as surgical prophylaxis was not associated with infection incidence within 30 days after LT, 180-day post-LT survival or CRO infection. Therefore, carbapenems are not superior to cephalosporin or piperacillin-tazobactam for perioperative antibiotic prophylaxis in LT recipients with a MELD score ≥30.

DETECTION OF CARBAPENEMASE PRODUCING ENTEROBACTERALES AMONG THE CLINICAL ISOLATES OF DIARRHOEAGENIC ESCHERICHIA COLI CAUSING ACUTE GASTROENTERITIS IN CHILDREN BELOW 2 YEARS IN TERTIARY CARE HOSPITAL OF NORTH EAST INDIA

Introduction: Enterobacterales that test resistant to at least one of the carbapenem antibiotics (ertapenem, meropenem, doripenem, or imipenem) are called Carbapenem resistant Enterobacterales (CRE) and if they produce a carbapenemase (an enzyme that can make them resistant to carbapenem antibiotics) they are called Carpenemase producing Enterobacterales (CPE). Children with CRE strains in fecal samples are considered as a high risk group by World Health Organization (WHO), which can spread CRE by intimate contact and travel. This cross-sectional study was conducted in the Department of Microbiology, RIMS, Imphal, Manipur, India fromMethods: Jan 2020 to Feb 2022. A total of 157 children under 2 years of age whose stool culture was positive for diarrhoeagenic Escherichia coli were included in the study. The modied carbapenem inactivation method (mCIM) has been done for detection of carbapenemase producers and the addition of EDTA in eCIM to further differentiate between serine and metallo-β-lactamase producers. Out of 157Result and Discussion: Diarrhoegenic E.coli (DEC) ,Carbapenem resistance was seen in 9 isolates i.e 5.7 %. Out of these 9 isolates, 3 were MBL producers tested by the phenotypic test mCIM and eCIM. All the three MBL producers carried bla NDM-1 gene. mCIM/eCIM assay is designed to simultaneously detect and distinguish the different types of carbapenemases. Carbapenemase genes are often located on plasmids that can be exchanged between Enterobacteriaceae and other Gram-negative bacteria. Carbapenem-resistant K. pneumoniae are currently more frequent and more likely to cause healthcareassociated outbreaks, carbapenem-resistant E. coli pose a greater risk for spread in the community. Conclusion: Screening for carbapenemase producer using mCIM and eCIM essay is important along with infection control measure such as active surveillance through rectal screening for CRE carriage on hospital admission, contact precautions, hand hygiene, patient isolation, environmental sanitation, case notication/agging, antibiotic restriction.

Novel Insight of Transcription Factor PtrA on Pathogenicity and Carbapenems Resistance in Pseudomonas aeruginosa.

IntroductionGlobally, Pseudomonas aeruginosa (PA) is emerging as a predominant nosocomial pathogen that often induces aggressive and even deadly infections. Pseudomonas type III repressor A (PtrA) can be activated specifically by copper ions and interacts with type-III transcriptional activator ExsA. This study aims to provide insight into the PtrA-mediated regulation of the pathogenicity and antibiotics resistance of PA.Methods and ResultsThe results of transcriptome sequencing analyses and real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) showed that PtrA plays a dual regulatory role in the virulence systems of PA: negatively regulates the type-III secretion system (T3SS) and positively regulates the quorum-sensing system (QS). The ptrA mutant attenuated extracellular virulence related to QS like pyocyanin, elastase, rhamnolipids, proteolytic activity, and biofilm production. According to adhesion and invasion experiments, PtrA can not only contribute to the adhesiveness but also the invasive of PA. Moreover, the PtrA-mediated regulation of PA pathogenicity was determined both in vivo and in vitro through cytotoxicity and Galleria mellonella survival experiments. In addition, apart from virulence, PtrA was found to influence the carbapenems resistance of PA. After deleting ptrA, the minimum inhibitory concentration (MIC) of carbapenems antibiotics was decreased by 2-fold, while a 2–8 fold increase was noted for the complemented strain.ConclusionOur findings establish that PtrA exerts a regulatory role in both pathogenicity and carbapenems resistance of PA. This work may shed light on a novel target for the clinical treatment of PA.

Class 1,2 Integron Genes Distribution in Pseudomonas aeruginosa Isolated from Clinical Specimens.

Pseudomonas aeruginosa, a Gram-negative human pathogen, P. aeruginosa is lone of the furthermost common hospital pathogens also is a chief concern, particularly in immune-compromised patients. The purpose of this study was to identify phenotypic and genotypic antibiotic resistance in Pseudomonas aeruginosa isolated from wound infection The bacterial isolates (30) were obtained from patients admitted to Mirjan Medical City in Babylon, Iraq (burns, wound unit). was identified biochemically and morphologically, and the isolates were subjected to standard bacteriological culturing processes on blood and MacConkey agar plates for 24-48 hours at 37oC for isolation and purification, Viteck 2 compact system confirmed the isolates and antibacterial sensitivity as well. These findings revealed that P.aeruginosa has a high rate of penicillin resistance, with a resistance rate of (100percent ) isolates. Advanced resistance to cephalosporin antibiotics was also found in resistant isolates of Cefoxitin, Ceftriaxone (75%), Ceftazidime, and cefepime (85%). For carbapenem antibiotics, had a high resistance rate (90percent). Aminoglycosides have variable resistance to Amikacin (60percent), Gentamicin (70 percent), and tobramycine resistant (90%). The findings revealed that all Pseudomonas aeruginosa isolates tested positive for Class 1,2 Integron resistance genes. with positive results (35%) for Int1 and 35% for Int2 (25%). This study found that P.aeruginosa has a high rate of resistance to Penicillins, Cephalosporin, Carbapenem and Aminoglycosides antibiotics.

Analysis of clinical characteristics, pathogen infection, and drug sensitivity of Marine injury patients: A cross-sectional study.

The infection rate is high in patients injured at sea, and because of the unique distribution of marine microorganisms, the infection is often not easily controlled effectively with the empirical application of antibiotics. This study aims to consider the clinical characteristics and pathogen infection and drug susceptibility of patients injured at sea. From 2019 to 2021, there were 635 patients injured at sea in Rizhao People’s Hospital. We assess the patient’s basic condition, while performing bacterial culture and drug susceptibility testing on wound exudate or pus from infected patients. Among the 635 patients injured at sea, 195 people were infected, and the infection rate was 30.71%. Infected patients are usually older, have longer prehospital visits, and have lower normal levels of red blood cells, hemoglobin, total protein, and albumin. The causes of injury in infected patients were mainly avulsion and puncture injuries, and the types of injuries were mainly bone fracture, vascular injury, and nerve injury. A total of 305 strains of pathogenic bacteria were cultured in 195 patients. Gram-negative bacteria accounted for 77.05% (235 strains), of which Proteus was the most. Gram-positive bacteria accounted for 22.95% (70 strains), of which Staphylococcus aureus was the most. Gram-negative bacilli were sensitive to aminoglycosides, lactam antibiotics, carbapenems antibiotics, sulfonamides, quinolones, fourth-generation cephalosporins, and antibacterial drugs containing enzyme inhibitors, while most of the bacteria were resistant to penicillins, first-generation cephalosporins, and second-generation cephalosporins. Gram-positive bacteria were sensitive to quinuptin/dafoptin, rifampicin, linezolid, gentamicin, tigacycline, and vancomycin but resistant to penicillin antibiotics. Due to the particularity of marine injuries, patients are prone to infection. Pathogen culture and drug sensitivity analysis play an important role in guiding antiinfective treatment for marine injured patients.

Occurrence and Characteristics of Carbapenem-Resistant Klebsiella pneumoniae Strains Isolated from Hospitalized Patients in Poland-A Single Centre Study.

The global emergence and spread of genes responsible for the production of ESBL (extended-spectrum beta-lactamases) and carbapenemases in Klebsiella pneumoniae isolates poses a serious threat to public health. The aim of this study was to retrospectively analyze the frequency of occurrence and drug resistance of selected alarm agents isolated from patients of the specialist hospital in Wrocław. A total of 13,528 clinical materials collected from patients of a specialist hospital in Wrocław were analyzed in the period from 1 January 2020 to 31 December 2020. Overall, 3894 bacterial strains were isolated from clinical materials, including 416 K. pneumoniae isolates. K. pneumoniae that showed resistance to ETP (ertapenem) and/or MEM (meropenem) were tested using phenotypic tests for the detection of KPC (carbapenemase-producing Klebsiella), MBL (metallo-β-lactamase) and OXA-48 (oxacilinase-48) carbapenemases. In the case of a positive or doubtful result of the phenotypic test, immunochromatographic tests and the CarbaNP test were performed. In total, 58 K. pneumoniae isolates resistant to 1 or more carbapenem antibiotics were isolated. Of the 58 strains, 16 (27.6%) were isolated from rectal swabs conducted on CPE (carbapenemase-producing Enterobacteriaceae) carriers. In the case of CRE (carbapenem-resistant Enterobacteriaceae) K. pneumoniae, carbapenemases were detected in 28/58 (48.3%) isolates. Notably, 23/28 K. pneumoniae isolates produced MBL/NDM (New Delhi metallo-β-lactamase) (82.1%), 5/28 produced VIM (Verona-intergon-encoded metallo-β-lactamase) (14.3%), and one produced MBL/NDM + OXA-48. Carbapenemases were detected in 13 of 16 (81.3%) carbapenem-resistant K. pneumoniae isolates derived from rectal swabs. The significant participation of CRE and CPE isolates in the infections proves the need to test patients admitted to hospital wards for their status as a CPE carrier in order to limit the emergence of new epidemic outbreaks.

Characterization of &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; isolated from positive samples of hemocultures and cerebrospinal fluid of children

Introduction. Infections of the bloodstream and central nervous system (CNS) caused by Pseudomonas aeruginosa are associated with a serious patient conditions and are often accompanied by high mortality.Aim. Molecular genetic characterization of P. aeruginosa isolated from positive samples of blood cultures and cerebrospinal fluid of patients under 18 years of age from intensive care units of hospitals.Materials and methods. We conducted a retrospective study of bacteremia and CNS infection cases associated with P. aeruginosa from 2014 to 2021. 24 clinical isolates of P. aeruginosa from positive blood cultures and CSF were analyzed. MICs of antibiotics were determined by serial microdilution in broth. Identification of the genes of carbapenemase was carried out using real-time PCR. Virulence genes were determined by PCR. Population diversity was assessed by MLST.Results. More than 70% of isolates showed resistance to carbapenem antibiotics. The phenotype of multiple drug resistance had 25% of the isolates. Extreme resistance was shown by 54% of isolates. The detection rate of metallo-β-lactamases (MBL) was 54%. Based on PCR data, 33% of the strains were found to have the ExoU type, and 67% had the ExoS type. According to MLST, 16 genotypes were identified. The structure was dominated by two sequence types ST654 (29%) and ST235 (12.5%). The structure of patients was dominated by children with surgical pathology — 16 cases, and there were eight somatic patients. Fatal outcome was observed in 28% of cases with bacteremia and CNS infection associated with P. aeruginosa.Conclusion. P. aeruginosa isolates from positive blood cultures and CSF samples are highly resistant to antibiotics; virulence genes were found in all isolates. Strains of high epidemic risk prevailed in the studied sample. More than a quarter of the described clinical cases had an unfavorable outcome.

Development of non-linear empirical models to estimate the abundance of carbapenem resistance genes during anaerobic digestion of wastewater sludge at mesophilic and thermophilic temperatures

Antibiotic resistance genes (ARGs) encode resistance against antibiotic drugs in bacteria. Carbapenem antibiotics are one of the last lines of defence against bacterial infections and carbapenem resistance genes (CRGs) accumulate in sludge during wastewater treatment. Anaerobic digestion is the most used method for sludge treatment, and it is important to know the abundance and removal of CRGs in anaerobically digested sludge before land application due to their public health relevance. However, gene quantification is time-consuming, costly, and sensitive to contamination. Previous research indicates a nexus between ARGs, solids, and nutrients in sludge. This study aimed to develop multiple regression models to estimate the abundance of CRGs from solids and nutrient concentrations in sludge, which are routinely measured for the monitoring and design of anaerobic digesters. Batch and continuous reactors were operated at mesophilic and thermophilic temperatures. Sludge samples were taken during digestion. CRGs (blaGES, blaOXA-48 and blaIMP-27), 16S rRNA and integron class 1 were quantified. Solids, chemical oxygen demand, orthophosphate and ammonia were measured as well. Anaerobic digestion reduced the quantity of the genes. First and second-order Kinetic rate constants (K1 and K2) were affected by temperature, and K1 absolute value followed the order blaOXA-48 > blaGES > Int 1. Multiple regression modelling was conducted in linear (LM) and non-linear (NLM) modes. This study presents the first NLM models to simulate the quantities of CRGs during anaerobic digestion of wastewater sludge at mesophilic and thermophilic temperatures. Adjusted R2 ranged 0.352–0.988 and 0.916–0.996 for the LM and NLM models, respectively. NLM models estimated the gene abundance more accurately than LM models, and they may be useful for researchers and practitioners to estimate the removal rate of target genes under different operational conditions.

Application of 1,1-ADEQUATE and DFT to correct 13 C misassignments of carbonyl chemical shifts for carbapenem antibiotics.

Prior to the development of sensitive proton-detected 2D NMR experiments, assigning 13 C signals could be a significant challenge, and mistakes have occurred even for prominent compound classes. In this study, 1,1-ADEQUATE data were used to unambiguously reassign the 13 C chemical shifts for the β-lactam carbonyl at the C-7 position and the proximal carboxylate at the C-10 position of the carbapenems, meropenem and imipenem. Density functional theory (DFT) was then investigated to provide sufficiently accurate 13 C chemical shift predictions, allowing for the carbonyl signal reassignment of thienamycin.