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UNLABELLED: Diazotrophic bacteria can reduce atmospheric nitrogen into ammonia enabling bioavailability of the essential element. Many diazotrophs closely associate with plant roots increasing nitrogen availability, acting as plant growth promoters. These associations have the potential to reduce the need for costly synthetic fertilizers if they could be engineered for agricultural applications. However, despite the importance of diazotrophic bacteria, genetic tools are poorly developed in a limited number of species, in turn narrowing the crops and root microbiomes that can be targeted. Here, we report optimized protocols and plasmids to manipulate phylogenetically diverse diazotrophs with the goal of enabling synthetic biology and genetic engineering. Three broad-host-range plasmids can be used across multiple diazotrophs, with the identification of one specific plasmid (containing origin of replication RK2 and a kanamycin resistance marker) showing the highest degree of compatibility across bacteria tested. We then demonstrated modular expression by testing seven promoters and eleven ribosomal binding sites using proxy fluorescent proteins. Finally, we tested four small molecule inducible systems to report expression in three diazotrophs and demonstrated genome editing in Klebsiella michiganensis M5al. ONE-SENTENCE SUMMARY: In this study, broad-host plasmids and synthetic genetic parts were leveraged to enable expression tools in a library of diazotrophic bacteria.

Background: Clinically important lineages in Klebsiella, especially those expressing multi-drug resistance (MDR), pose severe threats to public health worldwide. They arose from the co-evolution of the vertically inherited core genome and horizontal gene transfers by plasmids, which has not been systematically explored. Methods: We designed KleTy, which consists of dedicated typing schemes for both the core genome and plasmids in Klebsiella. We compared the performance of KleTy with many state-of-the-art pipelines using both simulated and real data. Results: Employing KleTy, we genotyped 33,272 Klebsiella genomes, categorizing them into 1773 distinct populations and predicting the presence of 87,410 plasmids from 837 clusters (PCs). Notably, Klebsiella is the center of the plasmid-exchange network within Enterobacteriaceae. Our results associated the international emergence of prevalent Klebsiella populations with only four carbapenem-resistance (CR) PCs, two hypervirulent PCs, and two hvCR-PCs encoding both carbapenemase and hypervirulence. Furthermore, we observed the ongoing international emergence of blaNDM, accompanied by the replacement of the previously dominant population, blaKPC-encoding HC1360_8 (CC258), during 2003–2018, with the emerging blaNDM-encoding HC1360_3 (CC147) thereafter. Additionally, expansions of hypervirulent carbapenem-resistant Klebsiella pneumoniae (hvCRKP) were evidenced in both populations, driven by plasmids of MDR-hypervirulence convergences. Conclusions: The study illuminates how the global genetic landscape of Klebsiella has been shaped by the co-evolution of both the core genome and the plasmids, underscoring the importance of surveillance and control of the dissemination of plasmids for curtailing the emergence of hvCRKPs. [ABSTRACT FROM AUTHOR]

Acupoint catgut embedding (ACE) is a safe and effective method for treating obesity. However, how it modulates intestinal flora and adiponectin remains unclear. We employed 16s rRNA sequencing technology to investigate ACE induced changes in intestinal flora and its association with adiponectin in subjects who received real (n = 41) and Sham (n = 41) stimulation. After treatment, the ACE group’s body weight, BMI, WC, HC, WHtR, and WHR were significantly lower than those of the Control group (p < .01), there was statistically significant differences in ADPN between the two groups (p < .01). ACE reduces the species abundance and increased the diversity and differences of the gut microbiota in perimenopausal patients with central obesity. Most notably, there was an increase in Kosakonia and Klebsiella after ACE treatment in the patients. Significant negatively correlations were found between body weight/waist circumference and adiponectin. Increases in Klebsiella and Kosakonia were positively correlated with adiponectin, and were negatively correlated with body weight/waist circumference. Our results showed increases in Klebsiella and Kosakonia were correlated with body weight/waist circumference and adiponectin. These findings suggest that ACE-induced weight loss is probably in part associated with increases in adiponectin, Klebsiella and Kosakonia.Trial re-registration: www.chictr.org.cn, ID: ChiCTR2400087718, Registration Date: August 2nd 2024. [ABSTRACT FROM AUTHOR]

Background: Antimicrobial resistance (AMR) can lead to fatal consequences. AMR genes carriage by phenotypically susceptible bacteria, such as Extended-Spectrum β-Lactamases (ESBL)s in Enterobacteriaceae , have potential implications for AMR spread and therapeutic outcomes. This phenomenon should be investigated. Methods: Positive blood cultures from hospitalized patients in a Malaysian tertiary center between April 2022 and March 2023 were reviewed. A total of 137 clinical isolates of Escherichia coli (E.coli), Klebsiella pneumoniae (K.pneumoniae), and Klebsiella oxytoca were included. The antibiotic susceptibility and ESBL phenotypes were determined by disk diffusion method and the identification of genotypes by multiplex polymerase chain reaction. The clinical characteristics and outcome information were extracted by reviewing patients' medical records to evaluate the clinical significance of the ESBL genotype-positive but phenotype-negative isolates in bacteremia. Results: All 137 isolates were positive for at least one genotype (bla CTX-M, n = 71, 51.8%; bla SHV, n = 87, 63.5%; bla TEM, n = 95, 69.3%; bla OXA-1, n = 38, 27.7%). While bla CTX-M was proportionately higher in the ESBL phenotype-positive isolates than ESBL phenotype-negative isolates (33/37, 89.2% vs 38/100, 38%; p < 0.001), more than half of those harboring bla CTX-M remained susceptible to third-generation cephalosporins (3GC). The sensitivity (Sen) of bla CTX-M for ESBL phenotypes prediction was 89.19% (95% confidence interval [CI], 74.58 - 96.97%); however, specificity (Sp) was low (46.47%; 95% CI 39.75 - 53.32). The patient characteristics were similar among 98 ESBL phenotype-negative cases, except that the non- bla CTX-M carrier group had significantly more renal impairment (0/37 vs 7/61, p = 0.043) and gastrointestinal sources of bacteremia (9/37 vs 27/61, p = 0.047). No differences were observed in infection severity, in-hospital mortality, and length of stay (LOS) between the bla CTX-M and non- bla CTX-M carrier groups. Conclusion: The current study provides insight into the gene carriage in E.coli and Klebsiella species clinical isolates, including bla CTX-M genotypes in antibiotic-susceptible strains from a Malaysian hospital. The ESBL encoding genotypes such as bla CTX-M presented substantially beyond one-third of the ESBL phenotype-negative or 3GC susceptible E.coli and K.pneumoniae isolated from bloodstream infection. Although clinical outcomes were not worsened with bla CTX-M genotype-positive but ESBL phenotype-negative isolates in bacteremia, the potential implications for AMR spread deserve further investigation. [ABSTRACT FROM AUTHOR]

Introduction: Klebsiella poses a significant global threat due to its high antibiotic resistance rate. In recent years, researchers have been seeking alternative antimicrobial agents, leading to the introduction of natural compounds such as monoterpenes, specifically thymol and carvacrol. This review aims to illustrate the potential antimicrobial, anti-biofilm, and synergistic traits of thymol and carvacrol in combat against Klebsiella. Methods: Searching PubMed, Scopus, and Web of Science, we reviewed available evidence on the antibacterial effects of thymol, carvacrol, or combined with other compounds against Klebsiella until May 2024. Reference checking was performed after the inclusion of studies. Minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), fractional inhibitory concentration (FIC), and anti-biofilm activity were gathered, and the MBC/MIC ratio was calculated to assess the bactericidal efficacy. Results: We retrieved 38 articles out of 2,652 studies screened. The gathered data assessed the anti-microbial activity of thymol, carvacrol, and both compounds in 17, 10, and 11 studies, respectively. The mean (± standard deviation) non-weighted MIC was 475.46 μg/mL (±509.95) out of 60 MIC for thymol and 279.26 μg/mL (±434.38) out of 68 MIC for carvacrol. Thymol and carvacrol showed anti-biofilm activities in the forms of disruption, inhibition, and mass reduction of biofilms. The MBC/MIC ratio was lower than 4 in 45 out of 47 cases, showing high bactericidal efficacy. FIC values were gathered for 68 combinations of thymol and carvacrol with other compounds, and they were mostly synergistic or additive. Conclusion: Thymol and carvacrol alone or in combination with other compounds, specifically known antibiotics, show great antimicrobial activity. [ABSTRACT FROM AUTHOR]

Klebsiella species are becoming a major global public health concern. In particular, the increase in multidrug‐resistant strains is a cause for concern. This study was aimed at determining the antibiotic susceptibility of two different isolates of Klebsiella quasipneumoniae subsp. similipneumoniae and determining the virulence characteristics and bacterial morphology under subminimum inhibitory concentrations (sub‐MICs) of antibiotics. In this study, two multidrug‐resistant K. quasipneumoniae subsp. similipneumoniae isolates were identified, one of which was clinical, and the other was isolated from freshwater. The MICs of the antibiotics meropenem, chloramphenicol, ciprofloxacin, and kanamycin were determined for these isolates. The effects of the sub‐MICs on the virulence and morphological characteristics of the bacteria were investigated in comparison with K. pneumoniae (ATCC 13883). The MICs of meropenem, chloramphenicol, ciprofloxacin, and kanamycin were 0.04, 20, 2, and 8 μg/mL in the clinical isolate; 0.2, 15, 5, and 2 μg/mL in the freshwater isolate; and 0.03, 3, 0.1, and 0.3 μg/mL for K. pneumoniae. The biofilm‐forming ability of K. quasipneumoniae subsp. similipneumoniae isolates decreased with antibiotic sub‐MICs. Siderophore activity increased only with MIC/4 of kanamycin and MIC/2 of chloramphenicol in the clinical isolate (p > 0.05). Furthermore, bacterial morphology and expression of virulence genes were affected differently by the sub‐MICs. This study showed that biofilm formation decreased and that the changes in bacterial morphology and expression of virulence genes were very different in the presence of 1/2 and 1/4 sub‐MIC antibiotics. [ABSTRACT FROM AUTHOR]

The genomic comparison of two Klebsiella michiganensis clinical isolates recovered from the same patient, one resistant to piperacillin-tazobactam and intermediate to cefotaxime, the other resistant to ceftazidime but susceptible to piperacillin-tazobactam, revealed one mutation in the blaOXY-1-24 gene accounting for a L169M substitution in the Ω loop. Cloning experiment in Escherichia coli demonstrated the contribution of this mutation to the hydrolysis spectrum extension towards ceftazidime and cefepime, whereas the resistance to piperacillin-tazobactam was reduced. To the best of our knowledge, this study shows for the first time that ceftazidime resistance can occur in vivo from OXY-1 precursor by structural alteration. [ABSTRACT FROM AUTHOR]

Isolating high-performance phenol degradation microorganisms with high salt tolerance and studying their resistance mechanisms are urgent issues. To address these issues, a typical bacteria (Klebsiella sp. YP-1) with high salt and phenol tolerance was isolated. Its strategies for resisting high salt and high phenol stress were studied. The results indicated that Klebsiella sp. YP-1 was able to degrade 1000 mg/L phenol within 44 h at 70 g/L NaCl, which was faster than most microorganisms reported in the literature. Lyxose secreted by Klebsiella sp. YP-1 played an important role on assisting Klebsiella sp. YP-1 to resist stress. Lyxose increased phenol degradation rate by microorganisms due to its protection on cell membrane. Quantum chemical calculation results indicated that lyxose was more likely attacked by free radical than cell membrane. In addition, lyxose could bind to the cell membrane through hydrogen bonds. Thus, lyxose prevented reactive oxygen species from harming cell membranes. Moreover, lyxose has broad protective effect on microbial cell membranes. This study provides a novel idea for microorganisms to resist oxidative stresses. [Display omitted] • A strain of Klebsiella sp. YP1 with high salt-phenol tolerance was isolated. • A novel strategy for Klebsiella sp. to resist stress was presented: secreting lyxose. • Lyxose improves the ability of microbe to degrade high salt-phenol wastewater. • The mechanism of lyxose protecting cell membrane was revealed for the first time. • Lyxose has broad protection effect on microbial cell membranes. [ABSTRACT FROM AUTHOR]

Aims Methods and Results Conclusions Hepatic inflammatory pseudotumours (IPTs) are nonneoplastic hepatic masses characterized by variably fibroblastic stroma and inflammatory infiltrate, hypothesized to arise as part of a response to infection or prior surgery. The aim of this study was to evaluate the clinicopathologic features and outcomes of biopsy‐proven hepatic IPT as well as other cases with IPT‐like histologic features.A database search at our institution identified cases with a pathologic diagnosis of hepatic IPT (n = 80) between 2000 and 2023. Histologic features (stromal quality, inflammatory cell components, granulomas, and necrosis) were evaluated. Past medical and surgical history, microbiologic studies, and outcomes were reviewed retrospectively. Patients frequently had a past medical history of malignancy (34%), biliary disease (15%), or prior intraabdominal surgery (24%), and often presented with multifocal hepatic lesions (36%). Variable inflammatory backgrounds were present, including histiocytic (36%), lymphoplasmacytic (34%), or neutrophilic (24%). Specific organisms were identified in 15% of cases, most commonly Klebsiella and Staphylococcus species. Most patients with available clinical follow‐up demonstrated radiologic resolution and/or had repeat negative biopsy; a minority of patients (8%) were subsequently diagnosed with neoplastic hepatic lesions. No significant association was seen between histologic features and the subsequent clinical or pathologic diagnosis of hepatic neoplastic lesions.Hepatic IPT is a heterogeneous entity that can present in a variety of clinical scenarios and show a wide morphologic spectrum. These lesions often regress spontaneously or with antibiotics. A subset of cases with hepatic IPT‐like histologic features were subsequently diagnosed with malignancy, emphasizing the need for continued follow‐up and repeat biopsy depending on clinical and radiologic features. [ABSTRACT FROM AUTHOR]

Background: Klebsiella quasipneumoniae (previously known as K. pneumoniae K6) strains are among the multidrug-resistant hypervirulent bacterial pathogens. Phage therapy can help treat infections caused by such pathogens. Here we report some aspects of virology and therapeutic potentials of vB_KquU_ 9K11K6, a bacteriophage that infects Klebsiella quasipneumoniae. Methods: K. quasipneumoniae (ATCC 700603) was used to screen wastewater lytic phages. The isolate vB_KquU_çKuK6 that consistently created large clear plaques was characterized using standard virological and molecular methods. Results: vB_KquU_çKuK6 has a complex capsid with an icosahedral head (~60 nm) and a slender tail (~140 nm x 10 nm). The phage has a 51% AT-rich linear dsDNA genome (51,251 bp) containing 121 open reading frames. The genome contains genes encoding spanin, endolysin, and holin proteins necessary for lytic infection and a recombinase gene possibly involved in lysogenic infection. vB_KquU_çKuK6 is stable at -80 to +67°C, pH 4-9, and brief exposure to one volume percent of chloroform. vB_KquU_çKuK6 has a narrow host range. Its lytic infection cycle involves a latency of 20 min and a burst size of 435 plaqueforming units. The phage can cause lysogenic infection, and the resulting lysogens are resistant to lytic infection by vB_KquU_çKuK6. vB_KquU_çKuK6 reduces the host cells' ability to form biofilm but fails to eliminate that ability. vB_KquU_çKuK6 demonstrates phage-antibiotic synergy and reduces the minimum inhibitory concentration of chloramphenicol and neomycin sulfate by about 8 folds. Conclusion: vB_KquU_çKuK6 cannot be directly used for phage therapy because it is a temperate bacteriophage. However, genetically modified strains of vB_KquU_çKuK6 alone or combined with antibiotics or other lytic Klebsiella phages can have therapeutic utilities in treating K. quasipneumoniae infections. [ABSTRACT FROM AUTHOR]

Infections with Enterobacterales (E) are increasingly difficult to treat due to antimicrobial resistance. After ceftriaxone replaced chloramphenicol (CHL) as empiric therapy for suspected sepsis in Malawi in 2004, extended-spectrum beta-lactamase (ESBL)-E rapidly emerged. Concurrently, resistance to CHL in Escherichia coli and Klebsiella spp. decreased, raising the possibility of CHL re-introduction. However, many phenotypically susceptible isolates still carry CHL acetyltransferase (cat) genes. To understand the molecular mechanisms and stability of this re-emerging CHL susceptibility we use a combination of genomics, phenotypic susceptibility assays, experimental evolution, and functional assays for CAT activity. Here, we show that of 840 Malawian E. coli and Klebsiella spp. isolates, 31% have discordant CHL susceptibility genotype–phenotype, and we select a subset of 42 isolates for in-depth analysis. Stable degradation of cat genes by insertion sequences leads to re-emergence of CHL susceptibility. Our study suggests that CHL could be reintroduced as a reserve agent for critically ill patients with ESBL-E infections in Malawi and similar settings and highlights the ongoing challenges in inferring antimicrobial resistance from sequence data. In this work, authors probe the molecular mechanism of re-emerging CHL susceptibility in Malawian Enterobacterales isolates, where they identify a stable truncation of resistance genes by insertion sequences. [ABSTRACT FROM AUTHOR]

Background Environmental disinfection is essential for reducing spread of healthcare-associated infections (HAIs). Previous studies report conflicting results regarding the effects of ultraviolet (UV) light in reducing infections. This trial evaluated the impact of adding pulsed-xenon UV (PX-UV) to standard terminal cleaning in reducing environmentally implicated HAIs (eiHAIs). Methods The Lowering the Acquisition of MDROs with Pulsed-xenon (LAMP) trial was conducted in 2 hospitals (15 inpatient wards) utilizing a cluster-randomized, controlled, double-blinded, interventional crossover trial comparing standard terminal cleaning followed by either PX-UV (intervention arm) or sham (control arm) disinfection. The primary outcome was incidence of eiHAIs from clinical microbiology tests on the fourth day of stay or later or within 3 days after discharge from the study unit. EiHAIs included clinical cultures positive for vancomycin-resistant enterococci, extended-spectrum beta-lactamase–producing Escherichia coli or Klebsiella pneumonia , methicillin-resistant Staphylococcus aureus , and Acinetobacter baumannii , and stool polymerase chain reaction (PCR) positive for Clostridiodes difficile. Results Between 18 May 2017 and 7 January 2020, 25 732 patients were included, with an incidence of 601 eiHAIs and 180 954 patient-days. There was no difference in the rate of eiHAIs in the intervention and sham arms (3.49 vs 3.17 infections/1000 patient-days, respectively; RR, 1.10; 95% CI,.94–1.29; P =.23). Study results were similar when stratified by eiHAI type, hospital, and unit type. Conclusions The LAMP study failed to demonstrate an effect of the addition of UV light disinfection following terminal cleaning on reductions in rates of eiHAIs. Further investigations targeting hospital environmental surfaces and the role of no-touch technology to reduce HAIs are needed. Clinical Trials Registration. ClinicalTrials.gov identifier: NCT03349268; R01HS024709. [ABSTRACT FROM AUTHOR]

Recycled manure solids (RMS) are dried cow dung processed using a manure dewatering machine and subsequently sun-dried to ~ 20% moisture. Benefits of RMS include abundant availability, low cost, and eco-friendliness, but its use as bedding material for cows is hindered by a moisture content that promotes microbial growth. This in vitro study evaluated impacts of calcium hydroxide (CH; 5 and 7.5%) and sodium hydrosulphate (SHS; 6 and 8%), independently and in combinations, at various depths of RMS, on physicochemical and microbial properties. The CH-treated groups had increased pH and reduced moisture on Day 0. Incorporating 7.5% CH + 6% SHS at 15–20 cm, and 7.5% CH + 8% SHS at all depths, effectively suppressed Escherichia coli and Klebsiella spp. Furthermore, a combination of 7.5% CH + 8% SHS at 20 cm inhibited coliform growth, whereas 7.5% CH with 6% SHS inhibited Streptococcus spp. In conclusion, a combination of 7.5% CH with either 6 or 8% SHS at a depth of 15 cm in RMS was particularly effective in controlling environmental mastitis-causing pathogens, specifically E. coli and Klebsiella spp. [ABSTRACT FROM AUTHOR]

This study aims to determine the pathogen organisms' profile and risk factors for infection after acute ischemic stroke (AIS) during the COVID-19 pandemic because of few studies. We conducted a retrospective cross-sectional study using the medical records of AIS inpatients at the National Brain Center Hospital Prof. Dr. dr. Mahar Mardjono, Jakarta, Indonesia, from 2020-2021. We found the species of pathogen organisms based on the positive growth of microbiological cultures of various specimens. Among 479 AIS patients, the infection prevalence was 12.3%. This study found the common pathogenic organisms were Gram-negative bacteria, and there were drugresistant strains in S. aureus, S. epidermidis, K. pneumoniae, and E. coli. The risk factors for infection in COVID-19-infected patients were pneumonia (OR 6.89, 95% CI 1.49-31.79, p = 0.013) and intensive care stay (OR 0.13, 95% CI 0.05-0.36, p < 0.001); meanwhile, in non-COVID-19 patients were HIV comorbidity (OR 1.55, 95% CI 1.18-2.06, p = 0.002), leukocytosis (OR 0.07, 95% CI 0.01-0.43, p = 0.004), use of CVC (OR 0.29, 95% CI 0.12-0.68, p = 0.005), use of steroids (OR 0.21, 95% CI 0.06-0.70, p = 0.011), and tracheostomy (OR 0.17, 95% CI 0.05-0.62, p = 0.007). To sum up, the growth of pathogenic organisms indicated that the prevalence of infections after AIS during the COVID-19 pandemic did not increase. The risk factor for infections depends on the characteristics of patients, whether they have COVID-19 or have not been infected. [ABSTRACT FROM AUTHOR]

To evaluate the effect of disposable cystoscopes on the rate of symptomatic urinary tract infections (UTI) following post-renal transplant cystoscopic stent removal. We performed a retrospective study of post-renal transplant cystoscopic stent removals in our outpatient clinic from March 2019 to March 2022. Our clinic converted to disposable cystoscopes in October 2021. All outpatient, phone, and portal encounters were reviewed for 30 days following the procedure. The primary outcome was the number of post-procedural symptomatic UTI within 30 days of the procedure. Symptomatic UTI was defined as fever, dysuria, or hematuria accompanied by a positive urine culture. A total of 323 patients had post-transplant stent removals including 123 with reusable scopes and 200 with disposable scopes. Around 1.6% (2/123) of patients with a reusable cystoscope experienced symptomatic UTI's. They had positive urine cultures for Escherichia coli and Klebsiella. 2.0% (4/200) of patients with a disposable cystoscopy had a symptomatic UTI. The 3 types of positive urine cultures they experienced were E Coli , Klebsiella, and Enterococcus. The conversion from reusable to disposable cystoscopes did not decrease symptomatic UTI following renal transplant stent removal. [ABSTRACT FROM AUTHOR]

This study continues surveillance of antimicrobial resistance associated with combat injuries in Ukraine. To compare species composition, antibiotic resistance profiles, and emergence of new resistance genes between 2014–2020 and 2022–2023. This was a retrospective multi-centre microbiological survey in Ukrainian hospitals. Antibiotic susceptibility, whole-genome sequencing and multi-locus sequence typing were conducted on 154 organisms obtained from 125 casualties between 2022 and 2023. The data revealed a predominance of Gram-negative bacteria, particularly Acinetobacter baumannii (35.7%), Pseudomonas aeruginosa (14.9%) and Klebsiella pneumoniae (20.7%). High levels of carbapenem resistance were observed among A. baumannii {meropenem 72.2% [39/54, 95% confidence interval (CI) 58.4–83.5]; imipenem 66.7% (36/54, 95% CI 52.5–78.9)}, K. pneumoniae [meropenem 90.6% (29/32, 95% CI 75.0–98.0); imipenem 81.2% (26/32, 95% CI 63.6–92.8)] and P. aeruginosa [meropenem 47.8% (11/23, 95% CI 26.8–69.4); imipenem 60.8% (14/23, 95% CI 38.5–80.3)] strains. A. baumannii sequence type (ST)-78 and ST-400 were prevalent from 2014 to 2020, while five strains of ST-1077 were newly identified in 2022–2023. P. aeruginosa strains showed diversity across 16 STs, with ST-773 increasing in frequency and new STs emerging, but lacking carbapenemase genes. K. pneumoniae exhibited increased genetic diversity over time, with three STs from 2014 to 2020 and six new STs, including bla NDM-1 , bla OXA-48 and bla KPC2 carriers, in 2022–2023. The prevalence of multi-drug-resistant isolates with STs associated with a high risk of global dissemination is increasing. [ABSTRACT FROM AUTHOR]

Background/Objectives: Urinary tract infection (UTI) is a prevalent microbial infection in medical practise, leading to significant patient morbidity and increased treatment costs, particularly in developing countries. This retrospective study, conducted at a tertiary care hospital in Dhaka, Bangladesh, aims to examine the antimicrobial resistance (AMR) patterns of uropathogens and evaluate whether these patterns are influenced by demographic factors such as gender, age, or patient status. Methods: Standard microbiological techniques were used to identify uropathogens, and AMR patterns were determined using the Kirby–Bauer disc diffusion method. Results: Out of 6549 urine samples, 1001 cultures were positive. The infection was more prevalent in females compared to males. The incidence of UTIs in children aged 0–10 years accounted for 12.59% of the total cases, with this age group also exhibiting the highest rate of polymicrobial infections. Among the bacterial uropathogens, 71.19% of isolates were multidrug resistant (MDR) and 84.27% were resistant to at least one antibiotic. Escherichia coli (n = 544, 73.90% MDR) and Klebsiella species (n = 143, 48.95% MDR) were the most common Gram-negative uropathogens, while Enterococcus species (n = 78, 94.87% MDR) was the predominant Gram-positive isolate in this study. Our results indicate that most uropathogens showed resistance against ceftazidime, followed by cefuroxime, trimethoprim-sulfamethoxazole, amoxicillin-clavulanate, and netilmicin. Moderate levels of resistance were observed against ciprofloxacin, levofloxacin, aztreonam, and cefpodoxime. Conclusions: Amikacin was observed to be effective against Gram-negative uropathogens, whereas cefixime was more active against Gram-positive microorganisms, such as Enterococcus species. Moreover, a principal coordinate analysis (PCoA) depicted no significant influence of gender, patient status, or age on AMR patterns. For the continued usefulness of most antibiotics, periodic analysis of the AMR patterns of uropathogens can help assess the rise of MDR bacteria, and therefore guide the selection of appropriate antibiotic treatment strategies. [ABSTRACT FROM AUTHOR]

Purpose: Children with congenital heart disease (CHD) from low- to middle-income countries (LMIC) are suspected to have a high prevalence of antibiotic-resistant microorganisms (ARMOs) carriage, but data are currently lacking. Carriage of ARMOs could impact the post-operative course in pediatric intensive care unit (PICU). The aim of the study was to assess the prevalence of ARMOs carriage in children with CHD from LMIC and its impact on post-operative outcomes. Methods: This was a retrospective monocentric study from 01/2019 to 12/2022. Included patients were children (0–18 years) from a LMIC admitted after CHD surgery and with AMRO screening performed the week before. Infections and post-operative evolution were compared based on ARMOs carriage status. Findings: Among 224 surgeries (median age 38.5 months (IQR 22–85.5)), ARMOs carriage was evidenced in 95 cases (42.4%). Main organisms isolated were Extended Spectrum Beta-Lactamase (ESBL) producing E. coli (75/224) 33.5%)) and ESBL-K. pneumoniae (30/224) 13.4%)). Median mechanical ventilation duration was 1 day (IQR 0–1), PICU stay 3 days (IQR 2–4) and hospital stay 6.5 days (IQR 5–10). A total of 17 infectious episodes occurred in 15 patients, mostly consisting in hospital-acquired pneumonia (HAP) (12/17). Only two infections were caused by a colonizing ARMO. Occurrence of infections and patients' outcome were similar between ARMO carriers and non-carriers. Higher use of carbapenems (6 (6.3%) vs 1 (0.8%), p = 0.04) and a trend to a higher use of vancomycin (14 (13.7%) vs 9 (6.9%), p = 0.04) in case of ARMOs carriage. Applying current guidelines, negative swab screening could have led to sparing most of empirical vancomycin therapy (11/12) for HAP based on current guidelines. Conclusion: Prevalence of AMROs carriage is high in children from LMIC and has a limited impact on patients' outcome. However, ARMOs carriage leads to higher consumption of antibiotics. Screening may help saving use of broad-spectrum antibiotic in non-carrier patients. [ABSTRACT FROM AUTHOR]

Objective: We aimed to determine the demographic data, mortality, and morbidity of early- and late-neonatal sepsis cases, the etiologic agents in these cases, and the antibiotic susceptibility of these agents. Methods: This study was conducted retrospectively in a tertiary neonatal intensive care unit (NICU). The demographic, clinical, and laboratory data of newborns diagnosed with culture-proven sepsis within 24 months were evaluated. Results: Two hundred and eleven culture data points belonging to 197 infants were evaluated. Forty percent of the infants had a history of premature birth. The most common clinical findings were respiratory distress and feeding intolerance. Coagulase-negative staphylococci (CoNS) were detected most frequently as early- and late-sepsis agents. The most common Gram-negative bacteria detected as late-sepsis agents were Klebsiella spp. and Escherichia coli (E. coli). The overall mortality rate was 10%. Conclusions: Neonatal sepsis continues to have high mortality rates in tertiary NICUs. CoNS was the most common agent, highlighting the importance of developing and maintaining personnel training and handwashing practices. It will be important to consider the resistance rates of Klebsiella spp., the most common Gram-negative agent in late-onset sepsis (LOS) cases, to commonly used antibiotics in empirical treatments. [ABSTRACT FROM AUTHOR]

Sterile insect technique (SIT) is a useful strategy for preventing and mitigative establishment of invasive insect species. SIT of the pest tephritid Mediterranean fruit fly, Ceratitis capitata (Wiedemann, 1824) WiedemannWiedemann , has been effective in preventing population establishment in vulnerable agricultural areas of the United States. However, irradiation-induced sterilization can have detrimental impacts resulting in reduced performance metrics. Mediterranean fruit fly males reared for SIT have been shown to have differences in their microbiomes relative to other population sources, which has been postulated to be a factor in how well flies compete with wild conspecifics. To identify baseline performance metrics on the effects of irradiation on the gut microbiome of mass-reared flies in Hawai'i, a study was performed to assess performance metrics and microbiome (bacterial 16S rRNA) variation across multiple timepoints. Mediterranean fruit fly pupae were selected from mass-reared trays intended for release, and paired samples were either irradiated or remained as controls and transported to the laboratory for evaluation. Irradiated flies exhibited fewer successful fliers, more rapid mortality rates, and were less active relative to control nonirradiated flies. Contrary to initial expectations, irradiation did not exert substantial impacts on the composition or diversity of bacterial reads. Samples were primarily comprised of sequences classified as Klebsiella and there were low levels of both read and taxonomic diversity relative to other 16S surveys of medfly. Although this study does not demonstrate a strong effect of irradiation alone on the Mediterranean fruit fly microbiome, there are several explanations for this discrepancy. [ABSTRACT FROM AUTHOR]

Introduction Case Description Discussion Conclusion This case examines the sonographic and clinical challenge of diagnosing a Klebsiella pnumoniae pyogenic liver abscess with systemic metastatic infection.The patient in this case study is an 81‐year‐old man who presented with intermittent rigors. Following radiological and clinical assessments, a Klebsiella pnumoniae pyogenic liver abscess, with evidence of systemic metastatic infection, was diagnosed. Sonographic features of the liver abscess were atypical for a cystic lesion and instead appeared as a solid mass, raising the possibility of malignancy. Treatment of intravenous ceftriaxone infusions resulted in full resolution of the liver lesion.The discussion criticises the terminology when describing hepatic lesions, which result from a Klebsiella pnumoniae infection. The term hepatic phlegmon is appropriate when a liver lesion caused by bacterial infection demonstrates a solid appearance on radiologic imaging. The term hepatic abscess is appropriate in cases where liver lesions caused by bacterial infection demonstrate a fluid filled core on radiologic imaging. Differentiation of these terms is important when treating the underlying lesion as a phlegmon, in contrast to an abscess, cannot be drained because it contains no pus. The variable sonographic appearances of Klebsiella pnumoniae pyogenic abscesses were also examined. Despite reported sonographic appearances in the literature, none are sufficient to distinguish a pyogenic liver abscess from malignancy without further investigation.Ultrasound operators should be aware of the variable sonographic appearances of a Klebsiella pnumoniae liver abscess and how these features, combined with non‐homogenous terminology, can obfuscate the correct diagnosis. [ABSTRACT FROM AUTHOR]

Background: Urinary tract infections (UTIs) are the second most common infection, affecting 150 million people each year worldwide. Enterobacteriaceae species expressing extended-spectrum β-lactamases (ESBLs) are on the rise across the globe and are becoming a severe problem in the therapeutic management of clinical cases of urinary tract infection. Knowledge of the prevalence and antibiogram profile of such isolates is essential to develop an appropriate treatment methodology. This study aimed to investigate the prevalence of Enterobacteriaceae isolates exhibiting ESBL and their selective oral antibiogram profile at the district general hospital, Polonnaruwa. Results: A total of 4386 urine specimens received to the Microbiology Laboratory during the study period. Among them, 1081 (24.6%) showed positive results for urine culture while 200/1081 specimens showed ESBL isolates. Out of the selected 200 specimen's majority (67.5%) of samples received from the In-Patient Department. There were 200 patients and reported that 115 (57.5%) were females and 85 (42.5%) were males. The majority (51%) of the patients belong to the age group of 55–74 years. Among the ESBLs positive specimens, the majority 74.5% (n = 149) identified organisms were E. coli followed by Klebsiella spp.17.5% (n = 35), Enterobacteriaceae 7% (n = 14) and only1% (n = 2) isolate of Proteus spp. Mecillinam (87.92%) and Nitrofurantoin (83.2%) showed higher effectiveness against E. coli. Nitrofurantoin showed the highest effectiveness against Klebsiella spp. (40%), other Enterobacteriaceae spp. (100%). Proteus spp. showed 100% effectiveness and resistance respectively against Ciprofloxacin, Cotrimoxazole and Nitrofurantoin. Conclusion: The most predominant ESBLs producing uro-pathogen was the E. coli in the study setting and E. coli had higher sensitivity rate against Mecillinam. Among currently used oral antibiotics Nitrofurantoin was the best choice for UTIs caused by ESBL producers. [ABSTRACT FROM AUTHOR]

Introduction: Urinary tract infections are most commonly caused by the uropathogenic bacterium Escherichia coli (UPEC), primarily affecting otherwise healthy women (50% of women will experience a urinary tract infection). These infections can become chronic and are highly recurrent. Objective: To compare the frequency of urinary infections caused by E. coli with those caused by other pathogenic bacteria. Materials and Methods: The study was conducted at the Biotest Laboratory Institute in Novi Sad. Data were obtained from archives for the period from January 2, 2023, to December 31, 2023. A total of 10,624 patients were included in the study. Of the total number of patients, 2,143 samples were positive for bacteria, representing 20% of the total number of analyses, while 8,499 samples were negative for bacterial presence in urine, representing 80% of the total number of patients tested. Results: Of the 2,143 samples positive for bacterial presence in urine: 1,740 samples had gram-negative bacteria isolated, and 403 samples had gram-positive bacteria isolated. This indicates that gram-negative bacteria were isolated in a large number of patients (81%), while gram-positive bacteria were isolated in 19% of patients. Of the 1,740 samples with isolated gram-negative bacteria: 1,392 samples had Escherichia coli, 224 samples had Klebsiella, 88 samples had Proteus, and 36 samples had other gram-negative bacteria. This shows that uropathogenic E. coli strains are by far the most common cause of urinary infections, representing 80% of the total number of gram-negative cultures. Klebsiella follows with 13%, Proteus with 5%, and other gram-negative bacteria with 2%. Conclusion: Urinary tract infections, encompassing various infectious etiologies, represent a significant threat to human health. Urinary tract infections are most commonly caused by the uropathogen Escherichia coli. [ABSTRACT FROM AUTHOR]

Background: Bacterial pneumonia is among the leading causes of morbidity and mortality worldwide. The extensive misuse and overuse of antibiotics observed during the Corona Virus Disease 2019 (COVID-19) pandemic may have changed the patterns of pathogens causing bacterial pneumonia and their antibiotic susceptibility profiles. This study was designed to establish the prevalence of culture-confirmed bacterial pneumonia and describe their antimicrobial susceptibility profile in adult patients who presented with signs and symptoms of lower respiratory tract infections (LRTIs) during the COVID-19 pandemic. Methodology: This hospital-based cross-sectional study was conducted from July 2021 to July 2022 at a zonal referral hospital and two district hospitals in Mwanza, Tanzania. Demographic and clinical data were collected using a standardized questionnaire. Sputum samples were processed by conventional culture followed by the identification of isolates and antibiotic susceptibility testing. Descriptive data analysis was performed using STATA version 15.0. Results: A total of 286 patients with a median age of 40 (IQR 29–60) years were enrolled in the study. More than half of the patients enrolled were females (52.4%, n = 150). The overall prevalence of bacterial pneumonia was 34.3% (n = 98). The majority of the bacterial pathogens isolated were Gram-negative bacteria (GNB) (61.2%, 60/98), with a predominance of Klebsiella spp., 38.8% (38/98), followed by Streptococcus pyogenes (21.4%, 21/98). Multi drug resistant (MDR) bacteria were detected in 72/98 (73.5%) of the isolates. The proportions of GNB-resistant strains were 60.0% (36/60) for ciprofloxacin, 60% (36/60) for amoxicillin, 60% (36/60) for amoxicillin, 68.3% (41/60) for trimethoprim-sulfamethoxazole and 58.3% (35/60) for ceftriaxone. Conclusion: One-third of the patients with signs and symptoms of LRTIs had laboratory-confirmed bacterial pneumonia with a predominance of Gram negative MDR bacteria. This calls for continuous antimicrobial resistance (AMR) surveillance and antimicrobial stewardship programs in the study setting and other settings in developing countries as important strategies for tackling AMR. [ABSTRACT FROM AUTHOR]

Background: The presence of contamination and microorganisms at any stage of processing renders a method unsafe, leading to a high risk of cross-transmission and cross-infection. Objective: The objective of this study was to assess the cleaning quality of aspirator instruments used in neurosurgical procedures. Methods: The experimental study was conducted at the materials and sterilization center, as well as the microbiology laboratory, of a philanthropic hospital in Brazil. A study protocol was implemented, which involved the analysis of 10 samples of Yasargil aspirators with varying dimensions. The samples were subjected to protein tests to detect the presence of organic matter and microbiological analysis. Descriptive statistics were used to analyze the data. Results: The results indicated that 40% of the instruments tested positive for protein after manual cleaning. Furthermore, after automated cleaning, samples showed an increased microbiological load, with Escherichia coli accounting for 20% and Klebsiella aerogenes for 10% of the identified microorganisms. Conclusion: This study provides evidence of failures in the cleaning process of healthcare products and highlights the presence of biofilm in the pipes, thereby compromising the drinking water quality standard. [ABSTRACT FROM AUTHOR]

Introduction: The increasing burden associated with drug-resistant bacteria is a global concern, and this issue has been highlighted across the world in the political arena through the Global Action Plan on antimicrobial resistance (AMR). Antimicrobial agents play a pivotal role and can be groundbreaking if chosen and administered correctly. Case description: Here, we describe a patient diagnosed with bacterial infection with an atypical susceptibility pattern. Conclusion: Imipenem-susceptible meropenem-resistant Klebsiella pneumoniae (ISMRK) is an unusual strain with varying acquired susceptibility. Comprehensive workup, knowledge, and appropriate treatment have been shown to lead to better outcome. [ABSTRACT FROM AUTHOR]

Background: The escalating resistance of uropathogens in pediatric febrile urinary tract infection (F-UTI) is a global concern. This study examined changing trends in F-UTI epidemiology and resistance patterns among Israeli pediatric inpatients over a decade. Methods: Demographic, clinical, and laboratory data for children between 3 months and 18 years old with febrile UTI from 2010 to 2021 were retrieved from electronic medical records. Results: A total of 761 cases of F-UTI were identified (702 females, mean age 43 months). Escherichia coli was the most common pathogen (85.9%), followed by Pseudomonas aeruginosa (3.5%) and Klebsiella pneumoniae (3.4%). Compared with the non-complicated UTI group, the complicated UTI group had significantly higher rates of Pseudomonas aeruginosa (5.3% vs. 1.0%, p = 0.002) and Klebsiella pneumoniae (4.6% vs. 1.6%, p = 0.03). Antibiotic resistance analysis revealed significant differences between the groups: resistance to cephalexin was higher in the complicated UTI group (19.3%) compared with the non-complicated UTI group (13.4%, p = 0.03). Notably, relatively low resistance rates were observed for ceftriaxone (4.4%) and gentamicin (6.0%). Over time, a significant decreasing trend in resistance to ampicillin was observed (slope = −0.0193, p = 0.011). No significant trends were found for trimethoprim–sulfamethoxazole, cephalexin, amoxicillin–clavulanic acid, ceftriaxone, and cefuroxime. Conclusions: Significant differences in pathogen distribution and resistance patterns between complicated UTI and non-complicated UTI groups highlight the need for continuous resistance monitoring and adherence to local guidelines. For the treatment of severe community F-UTI, ceftriaxone could be a reasonable option for first-onset F-UTI. Further studies are needed to implement antibiotic stewardship and optimize usage. [ABSTRACT FROM AUTHOR]

Background. In the context of increasing antimicrobial resistance (AMR), whole-genome sequencing (WGS) of bacteria is considered a highly accurate and comprehensive surveillance method for detecting and tracking the spread of resistant pathogens. Two primary sequencing technologies exist: short-read sequencing (50–300 base pairs) and long-read sequencing (thousands of base pairs). The former, based on Illumina sequencing platforms (ISPs), provides extensive coverage and high accuracy for detecting single nucleotide polymorphisms (SNPs) and small insertions/deletions, but is limited by its read length. The latter, based on platforms such as Oxford Nanopore Technologies (ONT), enables the assembly of genomes, particularly those with repetitive regions and structural variants, although its accuracy has historically been lower. Results. We performed a head-to-head comparison of these techniques to sequence the K. pneumoniae VS17 isolate, focusing on blaNDM resistance gene alleles in the context of a surveillance program. Discrepancies between the ISP (blaNDM-4 allele identified) and ONT (blaNDM-1 and blaNDM-5 alleles identified) were observed. Conjugation assays and Sanger sequencing, used as the gold standard, confirmed the validity of ONT results. This study demonstrates the importance of long-read or hybrid assemblies for accurate carbapenemase resistance gene identification and highlights the limitations of short reads in the context of gene duplications or multiple alleles. Conclusions. In this proof-of-concept study, we conclude that recent long-read sequencing technology may outperform standard short-read sequencing for the accurate identification of carbapenemase alleles. Such information is crucial given the rising prevalence of strains producing multiple carbapenemases, especially as WGS is increasingly used for epidemiological surveillance and infection control. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the microbiological quality of coconut water sold from street carts equipped with cooling coils or refrigerated at bakeries in the Grande Vitória Region, Brazil. Additionally, it assessed the phenotypic and genotypic antimicrobial resistance profiles of isolated enterobacteria. The results indicated that coconut water sold at street carts had lower microbiological quality compared to refrigerated samples, as evidenced by significantly higher counts of mesophilic microorganisms. Using MALDI-TOF, the following opportunistic pathogens were identified: Citrobacter freundii, Enterobacter bugandensis, E. kobei, E. roggenkampii, Klebsiella pneumoniae, and Kluyvera ascorbata. Three isolates—E. bugandensis, K. pneumoniae, and K. ascorbata—were classified as multidrug-resistant (MDR). Widespread resistance to β-lactams and cephalosporins was detected, and some isolates were resistant to quinolones, nitrofurans, and phosphonic acids. The gene blaCTX-M-2 was detected in C. freundii, E. bugandensis, E. kobei, and K. ascorbata. However, genes blaNDM, blaKPC, blaCMY-1, and blaCMY-2 were not detected in any isolate. The findings underscore the need to enhance good manufacturing practices in this sector to control the spread of antimicrobial resistance (AMR). To our knowledge, this is the first study documenting the presence of potentially pathogenic enterobacteria in coconut water samples and their associated phenotypic and genotypic AMR profiles. [ABSTRACT FROM AUTHOR]

Abstract Carbapenem-resistant Klebsiella pneumoniae (CRKp) infections continue to be an important cause of mortality. In this retrospective study, the effect of carbapenem or colistin resistance on mortality in Klebsiella pneumoniae bacteremia and combined meropenem + colistin administration in CRKp bacteremia was evaluated. In addition to that, a mathematical model is applied to explore the relationships between the resistance and mortality. A total of 139 adult patients diagnosed with K. pneumoniae bacteremia(73 carbapenem sensitive and 66 carbapenem resistant) between 01/01/2000 and 31/07/2019 were included in the study. The 30-day mortality in entire cohort were 19.4%. 30-day mortality was significantly higher in the carbapenem resistant-colistin sensitive group and in the carbapenem resistant-colistin resistant group compared to the carbapenem susceptible group. Meropenem + colistin combination was administered to 37 (95%) of carbapenem resistant–colistin sensitive (n = 39) and 25 (93%) of carbapenem resistant–colistin resistant patients(n = 27). Notably, mortality was not significantly affected regardless of whether CRKp was colistin sensitive and whether a high dose and prolonged infusion of meropenem was administered. Mortality is higher in carbapenem resistant Klebsiella pneumoniae bacteremia compared to carbapenem susceptible group. In cases of combined meropenem and colistin administration, high dose and prolonged infusion of meropenem is not superior to standard dose and infusion in both carbapenem resistant–colistin sensitive and carbapenem resistant–colistin resistant K. pneumoniae bacteremia.

• Sb(III) oxidation was mediated by AQDS and phenolic. • · OH, O 2 −• and SQ−• were found as dominant free radicals intermediates. • Oxidation was accelerated by free radical catalysis and electron transfer. • H 2 O 2 was dominant oxidant for Sb(III) oxidation in metabolites. • The secondary mineral mopungite formation sequesters part of Sb(V). Microbial oxidation and the mechanism of Sb(III) are key governing elements in biogeochemical cycling. A novel Sb oxidizing bacterium, Klebsiella aerogenes HC10, was attracted early and revealed that extracellular metabolites were the main fractions driving Sb oxidation. However, linkages between the extracellular metabolite driven Sb oxidation process and mechanism remain elusive. Here, model phenolic and quinone compounds, i.e., anthraquinone-2,6-disulfonate (AQDS) and hydroquinone (HYD), representing extracellular oxidants secreted by K. aerogenes HC10, were chosen to further study the Sb(III) oxidation mechanism. N 2 purging and free radical quenching showed that oxygen-induced oxidation accounted for 36.78% of Sb(III) in the metabolite reaction system, while hydroxyl free radicals (· OH) accounted for 15.52%. · OH and H 2 O 2 are the main driving factors for Sb oxidation. Radical quenching, methanol purification and electron paramagnetic resonance (EPR) analysis revealed that · OH, superoxide radical (O 2 •−) and semiquinone (SQ−•) were reactive intermediates of the phenolic induced oxidation process. Phenolic-induced ROS are one of the main oxidants in metabolites. Cyclic voltammetry (CV) showed that electron transfer of quinone also mediated Sb(III) oxidation. Part of Sb(V) was scavenged by the formation of the secondary Sb(V)-bearing mineral mopungite [NaSb(OH) 6 ] in the incubation system. Our study demonstrates the microbial role of oxidation detoxification and mineralization of Sb and provides scientific references for the biochemical remediation of Sb-contaminated soil. [Display omitted] [ABSTRACT FROM AUTHOR]

Introduction: The aim of this study was to describe a very rare case of endogenous bacterial endophthalmitis caused by Klebsiella pneumoniae in both eyes with a difference in the onset of symptoms of 14 months in an immunocompetent patient. Case Presentation: A 66-year-old immunocompetent man presented with asynchronous bilateral endogenous endophthalmitis produced by the K. pneumoniae bacterium at the starting point of a liver abscess after cholecystectomy surgery, causing endophthalmitis 1 year and 2 months apart between an eye and another. The first was diffuse anteroposterior endophthalmitis in the left eye that ended in visual loss and phthisis bulbi due to delayed initial diagnosis and established treatment, and the second was focal endophthalmitis in the right eye that preserved the organ and resulted in a vision of 20/20 due to early suspected diagnosis and rapid instituted treatment. Conclusion: To our knowledge, this is the first published case of a long asynchronous bilateral endogenous bacterial endophthalmitis caused by K. pneumoniae with a prolonged difference of 14 months in the onset of symptoms between one eye and another. This case is a vision-threatening ophthalmologic emergency that can be associated with life-threatening systemic morbidities. The early diagnosis of infection represents a challenge for clinicians, ophthalmologists, and microbiologists.

Abstract Background The human oral and nasal cavities can act as reservoirs for opportunistic pathogens capable of causing acute infection. These microbes asymptomatically colonize the human oral and nasal cavities which facilitates transmission within human populations via the environment, and they routinely possess clinically significant antibiotic resistance genes. Among these opportunistic pathogens, the Klebsiella genus stands out as a notable example, with its members frequently linked to nosocomial infections and multidrug resistance. As with many colonizing opportunistic pathogens, the essential transmission factors influencing the spread of Klebsiella species among both healthy and diseased individuals remain unclear. Results Here, we explored a possible explanation by investigating the ability of oral and nasal Klebsiella species to outcompete their native microbial community members under in vitro starvation conditions, which could be analogous to external hospital environments or the microenvironment of mechanical ventilators. When K. pneumoniae and K. aerogenes were present within a healthy human oral or nasal sample, the bacterial community composition shifted dramatically under starvation conditions and typically became enriched in Klebsiella species. Furthermore, introducing K. pneumoniae exogenously into a native microbial community lacking K. pneumoniae, even at low inoculum, led to repeated enrichment under starvation. Precise monitoring of K. pneumoniae within these communities undergoing starvation indicated rapid initial growth and prolonged viability compared to other members of the microbiome. K. pneumoniae strains isolated from healthy individuals’ oral and nasal cavities also exhibited resistance to multiple classes of antibiotics and were genetically similar to clinical and gut isolates. In addition, we found that in the absence of Klebsiella species, other understudied opportunistic pathogens, such as Peptostreptococcus, increased in relative abundance under starvation conditions. Conclusions Our findings establish an environmental and microbiome community circumstance that allows for the enrichment of Klebsiella species and other opportunistic pathogens. Klebsiella’s enrichment may hinge on its ability to quickly outgrow other members of the microbiome. The ability to outcompete other commensal bacteria and to persist under harsh environmental conditions could be an important factor that contributes to enhanced transmission in both commensal and pathogenic contexts. Video Abstract

Background. The outcome of complex treatment of implant-associated infection (IAI) depends on various factors, but one of the most important is the etiology of the inflammatory process. Treatment of orthopedic infection caused by Gram-negative microorganisms in general and representatives of the family Enterobacteriaceae in particular causes many difficulties, one of which is the rapid growth of antibiotic resistance. Aim of the study — to evaluate the factors affecting the course and prognosis of implant-associated infection caused by Klebsiella spp. Methods. We performed a retrospective analysis of case histories of 85 patients who underwent treatment of IAI caused by Klebsiella spp. in the clinical departments of the center from January 1, 2017 to December 31, 2021. According to the results of the telephone survey, the patients were divided into 2 main groups depending on the outcome of the 2-year follow-up period determined in accordance with the Delphi criteria. Results. It was found that the prognosis was significantly worsened by the number of sanitizing surgical interventions in the anamnesis (p = 0.022), the need for sanitizing intervention in the early postoperative period (p0.001) and the presence of Klebsiella spp. growth in postoperative culture tests (p = 0.002), hypoalbuminemia on 7-14 days after the surgery (p = 0.008). The administration of trimethoprim-sulfamethoxazole for the outpatient treament stage significantly improved the outcome (p = 0.038), which is most likely due to a high proportion of polymicrobial associations — 69.5%. Conclusions. There is a statistically significant direct relationship between the probability of an unfavorable treatment outcome of patients with IAI caused by Klebsiella spp. and the number of sanitizing surgical interventions in the anamnesis, low serum albumin (g/l) on 7-14 days after the operation, revision intervention in the early postoperative period, positive growth of Klebsiella spp. in postoperative culture tests. The probability of a favorable outcome was increased by the prescription of trimethoprim-sulfamethoxazole for oral administration at the outpatient stage.

Background: Invasive Klebsiella pneumoniae syndrome is a significant endemic disease in Taiwan. Intestinal colonization of virulent clones that cause this phenomenon has been demonstrated in asymptomatic adults. Comparisons of healthy adults and children with stool K. pneumoniae colonization have rarely been reported. We aimed to evaluate the frequency and abundance of K. pneumoniae in the stool of adults and children by stool microbiota analysis. Methods: Healthy volunteers and their children without antibiotic exposure within 3 months were recruited in a Taiwanese medical center. Stool samples were sent for gut microbiota analysis, using amplification of V3–V4 hypervariable regions of 16sRNA followed by high-throughput sequence. Rectal/stool swabs were sent for K. pneumoniae culture and identification by matrix-assisted laser desorption ionization–time-of-flight mass spectrometry (MALDI-TOF MS). Results: Fifty-five adults with a mean age of 46.9 years (range, 23.1–72.1 years) and 20 children with a mean age of 2.3 years (range, 0.9–5.8) were enrolled, and 29 adults and 6 children had positive K. pneumoniae swabs. Children had lower microbiota diversity than adults, including higher abundance of phylum Actinobacteria and Proteobacteria, and lower Bacteriodetes. For genus comparison, higher abundance of Escherichia, Streptococcus, Enterococcus and Bifidobacterium were found in children, but the composite abundance of Klebsiella in adults (median: 0.0156, range: 0–0.031) and in children (median: 0.0067, range: 0–0.043) were similar. Klebsiella abundance was significantly higher in participants with positive swabs (p

Background: cultivated and wild plants are used to treat different ailments. The Astragalus genus is found in temperate and dry climates; thus, it is found in Egypt and the arab world. Astragalus caprinus has a good amount of bioactive chemicals, which may help explain its therapeutic effects in reducing the risk of consequences from disease. Method: The phytochemical investigation of the herb and roots of Astragalus caprinus L. included the analytical characterization for the petroleum ether components by GC/MS, unsaponifiable matter (unsap. fraction), and fatty acids (FAME) investigation by GLC analysis. Main flavonoids were chromatographically isolated from ethyl acetate and n-butanol extracts. In vitro antimicrobial activity has been tested against the Gram-positive bacteria Staphylococcus aureus and Streptococcus mutans for different plant extracts, the Gram-negative bacteria Pseudomonas aeruginosa and Klebsiella pneumonia, the fungus Candida albicans and Aspergillus niger, and the Escherichia coli bacterium. Metabolite cytotoxicity was examined using the MTT assay against HepG-2 (human liver carcinoma) and MCF-7 (breast carcinoma). Results: Identifying the important components of the herb and root petroleum ether extracts was achieved. Using column chromatography, luteolin, cosmosiin (apigenin-7-O-glucoside), and cynaroside (luteolin-7-O-glucoside) were separated and identified using UV, NMR, and Mass Spectroscopy. Root extracts displayed potential antimicrobial activity against most of the tested pathogens. Both extracts (herb and roots) were active against the MCF-7 cell line and HepG-2 cell line with IC50 62.5 ± 0.64 and 72.4 ± 2.3 µg/ml, and 75.9 ± 2.5 and 96.8 ± 4.2 µg/ml, respectively. Conclusion: Astragalus caprinus seems to be a promising source of bioactive compounds that could potentially aid in preventing disease complications and address common health issues in developing countries. Moreover, the various parts of this plant could be utilized as natural raw materials for producing health-boosting products that could address common health issues in developing countries. [ABSTRACT FROM AUTHOR]

The antibiofilm and antimicrobial properties of tropical honey types including Malaysian stingless bee honey remain explicitly unexplored when compared with Apies honey. The antibiofilm and antimicrobial activities of the Malaysian Trigona honey were characterized with two stinging bee honey types (Centaurea hyalolepis and Citrus honeys) from Jordan. The antibiofilm and antimicrobial investigations were conducted on a set of seven microbial strains; five bacterial species of Pseudomonas aeruginosa ATCC 10145, Streptococcus pyogenes ATCC 19615, Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 13883, and two fungal strains Candida albicans ATCC 10231 and Candida krusei ATCC 14243. The antimicrobial investigations revealed a broad spectrum activity for Trigona honey against Gram-positive, Gram-negative, and fungal strains over the two honey types. One-way ANOVA showed a significant difference (p < 0.001) in the zone of inhibition ranging from 9 to 25 mm and minimum inhibition activity (MIC) ranged from 9.4–29.6% (w/v) against the microbial strains. Moreover, the addition of honey to established biofilms has induced a degradation activity in the biofilm mass. Two-way ANOVA showed a significant biofilm degradation proportion (p < 0.001) ranging from 1.3% to 91.3% following treatment with Trigona honey and the other honey types in relevance to the concentration ranging from 10% to 50% (w/v). Moreover, the antibiofilm activity was highly consistent with MIC affecting bacterial growth inhibition. In conclusion, a robust antimicrobial and antibiofilm activity for Trigona stingless bee honey over the stinging bee Centaurea hyalolepis and Citrus honeys is noticed which endows the usage of Trigona honey in the antimicrobial industry. [ABSTRACT FROM AUTHOR]

Two healthy Landrace pigs anaesthetized with propofol suffered rapid onset of fatal sepsis. Clinical signs included severe arterial hypotension, loss of peripheral oxygenation, low end-tidal CO2, clinical onset of pulmonary oedema and cardiac dysfunction. Gross and histopathological examination revealed loss of vascular integrity with severe lung oedema and congestion, haemorrhages in several organs and fluid leakage into body cavities. Large numbers of Gram-negative bacteria, primarily Klebsiella sp., were present in the anaesthetic infusion containing propofol and were also cultured from internal organs of both pigs. The propofol was likely contaminated by bacteria after inappropriate handling and storage in the operating room. This report illustrates the potential for severe nosocomial infection when applying propofol in animals and humans and may serve as a reminder of the importance of strict aseptic practice in general, and specifically in the handling of this anaesthetic agent. [ABSTRACT FROM AUTHOR]

Ibuprofen (IBU) is a significant contaminant frequently found in wastewater treatment plants due to its widespread use and limited removal during treatment processes. This leads to its discharge into the environment, causing considerable environmental concerns. The use of microorganisms has recently been recognized as a sustainable method for mitigating IBU contamination in wastewater. In this study, new bacteria capable of growing in a solid medium with IBU as the only carbon source and removing IBU from a liquid medium were isolated from environmental samples, including soil, marine, mine, and olive mill wastewater. Four bacterial strains, namely Klebsiella pneumoniae TIBU2.1, Klebsiella variicola LOIBU1.1, Pseudomonas aeruginosa LOIBU1.2, and Mycolicibacterium aubagnense HPB1.1, were identified through 16S rRNA gene sequencing. These strains demonstrated significant IBU removal efficiencies, ranging from 60 to 100% within 14 days, starting from an initial IBU concentration of 5 mg per litre. These bacteria have not been previously reported in the literature as IBU degraders, making this work a valuable contribution to further studies in the field of bioremediation in environments contaminated by IBU. Based on the IBU removal results, the most promising bacteria, K. pneumoniae TIBU2.1 and M. aubagnense HPB1.1, were selected for an in silico analysis to identify genes potentially involved in IBU biodegradation. Interestingly, in the tests with TIBU2.1, a peak of IBU transformation product(s) was detected by high‐performance liquid chromatography, while in the tests with HPB1.1, it was not detected. The emerging peak was analysed by liquid chromatography–mass spectrometry, indicating the presence of possible conjugates between intermediates of IBU biodegradation. The proteins encoded on their whole‐genome sequences were aligned with proteins involved in an IBU‐degrading pathway reported in bacteria with respective catabolic genes. The analysis indicated that strain HPB1.1 possesses genes encoding proteins similar to most enzymes reported associated with the IBU metabolic pathways used as reference bacteria, while strain TIBU2.1 has genes encoding proteins similar to enzymes involved in both the upper and the lower part of that pathway. Notably, in the tests with the strain having more candidate genes encoding IBU‐catabolic enzymes, no IBU transformation products were detected, while in the tests with the strain having fewer of these genes, detection occurred. [ABSTRACT FROM AUTHOR]

We study the bacterial respiration through the numerical solution of the Fairen–Velarde coupled nonlinear differential equations. The instantaneous concentrations of the oxygen and the nutrients are computed. The fixed point solution and the stable limit cycle are found in different parameter ranges as predicted by the linearized differential equations. In a specified range of parameters, it is observed that the system spends some time near the stable limit cycle and eventually reaches the stable fixed point. This metastability has been investigated systematically. Interestingly, it is observed that the system exhibits two distinctly different time scales in reaching the stable fixed points. The slow time scale of the metastable lifetime near the stable limit cycle and a fast time scale (after leaving the zone of limit cycle) in rushing towards the stable fixed point. The gross residence time, near the limit cycle (described by a slow time scale), can be reduced by varying the concentrations of nutrients. This idea can be used to control the harmful metastable lifespan of active bacteria. [ABSTRACT FROM AUTHOR]

Objectives: Rhinoscleroma is classically described as a chronic granulomatous disease caused by Klebsiella rhinoscleromatis which primarily affects the nose and nasopharynx. When present, tracheal manifestations will be seen late in the disease course rather than on initial presentation. We describe a rare case of nonendemic rhinoscleroma that presented with tracheal lesions as an initial manifestation of disease. Methods: Case report and literature review. Results: An 88‐year‐old male presented with longstanding dysphonia. Flexible laryngoscopy demonstrated a septal perforation and diffuse glottic lesions. CT neck demonstrated a nonobstructive polypoid tracheal lesion and mucosal thickening of the paranasal sinuses. Biopsy confirmed an atypical lympho‐histiocytic proliferation and microorganisms within macrophages on Grocott methenamine silver and Steiner stains consistent with rhinoscleroma. He was referred for rheumatology and pulmonology consultation. Conclusion: Systemic diseases rarely affect the trachea, and even less frequently is a tracheal lesion identified as the initial manifestation of disease. The most common systemic diseases that affect the trachea include relapsing polychondritis, granulomatosis with polyangiitis, amyloidosis, and inflammatory bowel disease. The literature surrounding nasolaryngotracheal rhinoscleroma is limited, especially in nonendemic areas. It is necessary to include unusual etiologies of airway lesions in the differential diagnosis, which warrants comprehensive airway evaluation including biopsy. Level of Evidence: 4. [ABSTRACT FROM AUTHOR]

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Aims: The emergence of antibiotics resistance in Acinetobacter baumannii and Klebsiella pneumoniae threatens the efficacy of the treatment regime. This study was carried out to evaluate the antibiotic resistance and phylogenetic relatedness in the metallo-β-lactamase (MBL) producers K. pneumoniae and A. baumannii. Methodology and results: In the current study, nine clinical samples from inpatients at Lady Reading Hospital (LRH) were taken from the City Medical Laboratory, Peshawar (Pakistan). Morphological, biochemical and molecular identification of the bacterial strains showed that 6 isolates were K. pneumoniae while 3 were A. baumannii. The phylogenetic tree was constructed using software Finch TV, Codon code aligner and Mega X. Antibiotic susceptibility profiles showed that 89% of isolates were multidrug-resistant (MDR). Sixty-seven (67%) of K. pneumoniae isolates were positive for the Modified Hodge test (MHT), Combined-disc test (CDT) and MBL test. The minimum inhibitory concentration (MIC) of metallo-β-lactamase positive strains were recorded up to 256 μg/mL. MIC values for K. pneumoniae were relatively higher as compared to A. baumannii. Conclusion, significance and impact of study: The presence of a high percentage of MBL producer multidrugresistant K. pneumoniae and A. baumannii isolates reported in this study may indicate a major problem for clinicians by complicating the treatment of infections in hospital settings and need immediate attention. [ABSTRACT FROM AUTHOR]

Background: Neonatal sepsis is a critical health issue worldwide, with high morbidity and mortality rates, especially in developing countries. Effective management of neonatal sepsis includes timely administration of appropriate antibiotics based on antimicrobial susceptibility patterns. Aims and Objectives: This cross-sectional study aimed to investigate the choice of empiric and definitive antibiotics in neonatal septicemia, assess bacterial susceptibility patterns, and correlation with clinical outcomes, and highlight the importance of antimicrobial stewardship. Materials and Methods: The study was conducted at a tertiary care center and enrolled neonates diagnosed with septicemia over 2 months. Blood cultures were obtained, and antibiotics were initiated empirically based on local guidelines. Definitive antibiotics were started after culture sensitivity reports and compared with empiric treatment for clinical outcomes and bacteriological profiles were analyzed with appropriate statistical methods. Results: In this study, Piperacillin-Tazobactam and Amikacin were used as empiric treatment in all cases. Meropenem showed high sensitivity in definitive treatment post-culture. No significant difference between empiric and definitive treatment groups was observed in clinical outcomes. Conclusion: This study found no significant difference in clinical outcomes between empiric and definitive antibiotic regimens for neonatal sepsis. However, high meropenem susceptibility suggests the potential for more targeted therapy. Regularly updating local antibiogram, data remain crucial to optimize empiric treatment and combat resistance in this vulnerable population. [ABSTRACT FROM AUTHOR]

To provide a scientific basis for controlling mulberry bacterial blight in Bazhong, Sichuan, China (BSC), this study aimed to isolate and purify pathogenic bacteria from diseased branches of mulberry trees in the region and to clarify their taxonomic status using morphological observation, physiological and biochemical detection, molecular-level identification, and the construction of a phylogenetic tree. A total of 218 bacterial strains were isolated from samples of diseased mulberry branches. Of these, 7 strains were identified as pathogenic bacteria based on pathogenicity tests conducted in accordance with Koch's postulates. Preliminary findings from the analysis of the 16S rRNA sequence indicated that the 7 pathogenic bacteria are members of Klebsiella spp. Morphological observation revealed that the pathogenic bacteria were oval-shaped and had capsules but no spores. They could secrete pectinase, cellulase, and protease and were able to utilize D-glucose, D-mannose, D-maltose, and D-Cellobiose. The 7 strains of pathogenic bacteria exhibited the highest homology with Klebsiella oxytoca. This study identifies Klebsiella oxytoca as the causative agent of mulberry bacterial blight in BSC, laying the foundation for the prevention and control of this pathogen and further investigation into its pathogenic mechanism. Supplementary Material Supplementary Material File [ABSTRACT FROM AUTHOR]

The increase in bacterial resistance is currently a global burden for the health care system. In order to evaluate the resistance rates of several bacteria from the most encountered cultures in clinical practice, we performed a retrospective analysis of all of the positive cultures from the year 2021 in a tertiary care hospital in Romania. Our analysis captured 3299 positive cultures. The median age of the patients was 62 years (IQR: 41–71 years old) with a slight predominance among females (53.1%). Overall, the most common cultures were urocultures, wound secretion cultures and blood cultures, and the most common identified bacteria were Escherichia coli, Staphylococcus aureus and Klebsiella spp. Positive cultures with the highest resistance rates were found in the bronchial aspirate cultures, catheter tip cultures, urocultures and blood cultures. Escherichia coli (n = 996) had the highest resistance to ampicillin (19.8%) and trimetoprim-sulfametoxazole (16.4%), while Staphylococcus aureus (n = 698) presented the highest resistance rates to clindamycin (27.4%) and oxaciline (19.7%). Klebsiella (n = 481) presented the highest resistance rates to piperaciline-tazobactam (25.2%) and ampicillin (20.4%), whereas Acinetobacter baumanii (n = 123) presented a resistance rate of more than 50% to carbapenems, gentamicin, ciprofloxacin and ceftazidime. The aim of our study was to identify bacterial resistance rates in order to provide updated clinical data to guide physicians in choosing the best empirical antibiotic treatment, especially in the west part of Romania. [ABSTRACT FROM AUTHOR]

The common intestinal pathogen Klebsiella pneumoniae (K. pneumoniae) is one of the leading causes of fatal superbug infections that can resist the effects of commonly prescribed medicines. The uncontrolled use or misuse of antibiotics has increased the prevalence of drug-resistant K. pneumoniae strains in the environment. In the quest to search for alternative therapeutics for treating these drug-resistant infections, bacteriophages (bacterial viruses) emerged as potential candidates for in phage therapy against Klebsiella. The effective formulation of phage therapy against drug-resistant Klebsiella infections demands thorough characterization and screening of many bacteriophages. To contribute effectively to the formulation of successful phage therapy against superbug infections by K. pneumoniae, this study includes the isolation and characterization of a novel lytic bacteriophage MKP-1 to consider its potential to be used as therapeutics in treating drug-resistant Klebsiella infections. Morphologically, having a capsid attached to a long non-contractile tail, it was found to be a siphovirus that belongs to the class Caudoviricetes and showed infectivity against different strains of the target host bacterium. Comparatively, this double-stranded DNA phage has a large burst size and is quite stable in various physiological conditions. More interestingly, it has the potential to degrade the tough biofilms formed by K. pneumoniae (Klebsiella pneumoniae subsp. pneumoniae (Schroeter) Trevisan [ATCC 15380]) significantly. Thus, the following study would contribute effectively to considering phage MKP-1 as a potential candidate for phage therapy against Klebsiella infection. [ABSTRACT FROM AUTHOR]

Background: (R,R)-2,3-butanediol (BDO) is employed in a variety of applications and is gaining prominence due to its unique physicochemical features. The use of glycerol as a carbon source for 2,3-BDO production in Klebsiella pneumoniae has been limited, since 1,3-propanediol (PDO) is generated during glycerol fermentation. Results: In this study, the inactivation of the budC gene in K. pneumoniae increased the production rate of (R,R)-2,3-BDO from 21.92 ± 2.10 to 92.05 ± 1.20%. The major isomer form of K. pneumoniae (meso-2,3-BDO) was shifted to (R,R)-2,3-BDO. The purity of (R,R)-2,3-BDO was examined by agitation speed, and 98.54% of (R,R)-2,3-BDO was obtained at 500 rpm. However, as the cultivation period got longer, the purity of (R,R)-2,3-BDO declined. For this problem, a two-step agitation speed control strategy (adjusted from 500 to 400 rpm after 24 h) and over-expression of the dhaD gene involved in (R,R)-2,3-BDO biosynthesis were used. Nevertheless, the purity of (R,R)-2,3-BDO still gradually decreased over time. Finally, when pure glycerol was replaced with crude glycerol, the titer of 89.47 g/L of (R,R)-2,3-BDO (1.69 g/L of meso-2,3-BDO), productivity of 1.24 g/L/h, and yield of 0.35 g/g consumed crude glycerol was achieved while maintaining a purity of 98% or higher. Conclusions: This study is meaningful in that it demonstrated the highest production and productivity among studies in that produced (R,R)-2,3-BDO with a high purity in Klebsiella sp. strains. In addition, to the best of our knowledge, this is the first study to produce (R,R)-2,3-BDO using glycerol as the sole carbon source. [ABSTRACT FROM AUTHOR]

Background Little is known about the risk of progression from carbapenemase-producing Enterobacterales (CPE) carriage to CPE bloodstream infection (BSI) outside of high-risk settings. We aimed to determine the incidence of CPE BSI among CPE carriers and to assess whether the incidence differs by carbapenemase, species, and setting. Methods We conducted a nationwide population-based retrospective cohort study using national databases. The cohort consisted of all patients in Israel with CPE detected by screening from 1 January 2020 to 10 October 2022. We calculated the cumulative incidence of CPE BSI within 1 year among CPE carriers. We used a competing-risks model with BSI as the outcome and death as the competing risk. Results The study included 6828 CPE carriers. The cumulative incidence of CPE BSI was 2.4% (95% confidence interval [CI], 2.1–2.8). Compared with Klebsiella pneumoniae carbapenemase (KPC), the subhazard of BSI was lower for New Delhi metallo-β-lactamase (NDM) (adjusted subhazard ratio [aSHR], 0.72; 95% CI,.49–1.05) and oxacillinase-48-like (OXA-48-like) (aSHR, 0.60; 95% CI,.32–1.12) but these differences did not reach statistical significance. Compared with K. pneumoniae , the subhazard of BSI was lower for carriers of carbapenemase-producing Escherichia coli (aSHR, 0.33; 95% CI,.21–.52). The subhazard of BSI was higher among patients with CPE carriage first detected in intensive care units (aSHR, 2.10; 95% CI, 1.27–3.49) or oncology/hematology wards (aSHR, 3.95; 95% CI, 2.51–6.22) compared with medical wards. Conclusions The risk of CPE BSI among CPE carriers is lower than previously reported in studies that focused on high-risk patients and settings. The risk of BSI differs significantly by bacterial species and setting, but not by carbapenemase. [ABSTRACT FROM AUTHOR]

Gram-negative antibiotic resistance continues to grow as a global problem due to the evolution and spread of β-lactamases. The early β-lactamase inhibitors (BLIs) are characterized by spectra limited to class A β-lactamases and ineffective against carbapenemases and most extended spectrum β-lactamases. In order to address this therapeutic need, newer BLIs were developed with the goal of treating carbapenemase producing, carbapenem resistant organisms (CRO), specifically targeting the Klebsiella pneumoniae carbapenemase (KPC). These BL/BLI combination drugs, avibactam/avibactam, meropenem/vaborbactam, and imipenem/relebactam, have proven to be indispensable tools in this effort. However, non-KPC mechanisms of resistance are rising in prevalence and increasingly challenging to treat. It is critical for clinicians to understand the unique spectra of these BL/BLIs with respect to non-KPC CRO. In Part 1of this 2-part series, we describe the non-KPC attributes of the newer BL/BLIs with a focus on utility against Enterobacterales and Pseudomonas aeruginosa. [ABSTRACT FROM AUTHOR]