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Abstract Background The emergence of multi-drug-resistant Klebsiella pneumoniae (MDR-KP) represents a serious clinical health concern. Antibiotic resistance and virulence interactions play a significant role in the pathogenesis of K. pneumoniae infections. Therefore, tracking the clinical resistome and virulome through monitoring antibiotic resistance genes (ARG) and virulence factors in the bacterial genome using computational analysis tools is critical for predicting the next epidemic. Methods In the current study, one hundred extended spectrum β-lactamase (ESBL)-producing clinical isolates were collected from Mansoura University Hospital, Egypt, in a six-month period from January to June 2022. One isolate was selected due to the high resistance phenotype, and the genetic features of MDR-KP recovered from hospitalized patient were investigated. Otherwise, the susceptibility to 25 antimicrobials was determined using the DL Antimicrobial Susceptibility Testing (AST) system. Whole genome sequencing (WGS) using Illumina NovaSeq 6000 was employed to provide genomic insights into K. pneumoniae WSF99 clinical isolate. Results The isolate K. pneumoniae WSF99 was phenotypically resistant to the antibiotics under investigation via antibiotic susceptibility testing. WGS analysis revealed that WSF99 total genome length was 5.7 Mb with an estimated 5,718 protein-coding genes and a G + C content of 56.98 mol%. Additionally, the allelic profile of the WSF99 isolate was allocated to the high-risk clone ST147. Furthermore, diverse antibiotic resistance genes were determined in the genome that explain the high-level resistance phenotypes. Several β-lactamase genes, including bla CTX−M−15, bla TEM−1, bla TEM−12, bla SHV−11, bla SHV−67, and bla OXA−9, were detected in the WSF99 isolate. Moreover, a single carbapenemase gene, bla NDM−5, was predicted in the genome, positioned within a mobile cassette. In addition, other resistance genes were predicted in the genome including, aac(6’)-Ib, aph(3’)-VI, sul1, sul2, fosA, aadA, arr-2, qnrS1, tetA and tetC. Four plasmid replicons CoIRNAI, IncFIB(K), IncFIB(pQil), and IncR were predicted in the genome. The draft genome analysis revealed the occurrence of genetic mobile elements positioned around the ARGs, suggesting the ease of dissemination via horizontal gene transfer. Conclusions This study reports a comprehensive pathogenomic analysis of MDR-KP isolated from a hospitalized patient. These findings could be relevant for future studies investigating the diversity of antimicrobial resistance and virulence in Egypt.

Background: The emergence of multi-drug-resistant Klebsiella pneumoniae (MDR-KP) represents a serious clinical health concern. Antibiotic resistance and virulence interactions play a significant role in the pathogenesis of K. pneumoniae infections. Therefore, tracking the clinical resistome and virulome through monitoring antibiotic resistance genes (ARG) and virulence factors in the bacterial genome using computational analysis tools is critical for predicting the next epidemic. Methods: In the current study, one hundred extended spectrum β-lactamase (ESBL)-producing clinical isolates were collected from Mansoura University Hospital, Egypt, in a six-month period from January to June 2022. One isolate was selected due to the high resistance phenotype, and the genetic features of MDR-KP recovered from hospitalized patient were investigated. Otherwise, the susceptibility to 25 antimicrobials was determined using the DL Antimicrobial Susceptibility Testing (AST) system. Whole genome sequencing (WGS) using Illumina NovaSeq 6000 was employed to provide genomic insights into K. pneumoniae WSF99 clinical isolate. Results: The isolate K. pneumoniae WSF99 was phenotypically resistant to the antibiotics under investigation via antibiotic susceptibility testing. WGS analysis revealed that WSF99 total genome length was 5.7 Mb with an estimated 5,718 protein-coding genes and a G + C content of 56.98 mol%. Additionally, the allelic profile of the WSF99 isolate was allocated to the high-risk clone ST147. Furthermore, diverse antibiotic resistance genes were determined in the genome that explain the high-level resistance phenotypes. Several β-lactamase genes, including blaCTX−M−15, blaTEM−1, blaTEM−12, blaSHV−11, blaSHV−67, and blaOXA−9, were detected in the WSF99 isolate. Moreover, a single carbapenemase gene, blaNDM−5, was predicted in the genome, positioned within a mobile cassette. In addition, other resistance genes were predicted in the genome including, aac(6')-Ib, aph(3')-VI, sul1, sul2, fosA, aadA, arr-2, qnrS1, tetA and tetC. Four plasmid replicons CoIRNAI, IncFIB(K), IncFIB(pQil), and IncR were predicted in the genome. The draft genome analysis revealed the occurrence of genetic mobile elements positioned around the ARGs, suggesting the ease of dissemination via horizontal gene transfer. Conclusions: This study reports a comprehensive pathogenomic analysis of MDR-KP isolated from a hospitalized patient. These findings could be relevant for future studies investigating the diversity of antimicrobial resistance and virulence in Egypt. [ABSTRACT FROM AUTHOR]

Abstract Background In recent years, there has been a growing interest in phage therapy as an effective therapeutic tool against colibacillosis caused by avian pathogenic Escherichia coli (APEC) which resulted from the increasing number of multidrug resistant (MDR) APEC strains. Methods In the present study, we reported the characterization of a new lytic bacteriophage (Escherichia phage AG- MK-2022. Basu) isolated from poultry slaughterhouse wastewater. In addition, the in vitro bacteriolytic activity of the newly isolated phage (Escherichia phage AG- MK-2022. Basu) and the Escherichia phage VaT-2019a isolate PE17 (GenBank: MK353636.1) were assessed against MDR- APEC strains (n = 100) isolated from broiler chickens with clinical signs of colibacillosis. Results Escherichia phage AG- MK-2022. Basu belongs to the Myoviridae family and exhibits a broad host range. Furthermore, the phage showed stability under a wide range of temperatures, pH values and different concentrations of NaCl. Genome analysis of the Escherichia phage AG- MK-2022. Basu revealed that the phage possesses no antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and any E. coli virulence associated genes. In vitro bacterial challenge tests demonstrated that two phages, the Escherichia phage VaT-2019a isolate PE17 and the Escherichia phage AG- MK-2022. Basu exhibited high bactericidal activity against APEC strains and lysed 95% of the tested APEC strains. Conclusions The current study findings indicate that both phages could be suggested as safe biocontrol agents and alternatives to antibiotics for controlling MDR-APEC strains isolated from broilers.

Background: In recent years, there has been a growing interest in phage therapy as an effective therapeutic tool against colibacillosis caused by avian pathogenic Escherichia coli (APEC) which resulted from the increasing number of multidrug resistant (MDR) APEC strains. Methods: In the present study, we reported the characterization of a new lytic bacteriophage (Escherichia phage AG- MK-2022. Basu) isolated from poultry slaughterhouse wastewater. In addition, the in vitro bacteriolytic activity of the newly isolated phage (Escherichia phage AG- MK-2022. Basu) and the Escherichia phage VaT-2019a isolate PE17 (GenBank: MK353636.1) were assessed against MDR- APEC strains (n = 100) isolated from broiler chickens with clinical signs of colibacillosis. Results : Escherichia phage AG- MK-2022. Basu belongs to the Myoviridae family and exhibits a broad host range. Furthermore, the phage showed stability under a wide range of temperatures, pH values and different concentrations of NaCl. Genome analysis of the Escherichia phage AG- MK-2022. Basu revealed that the phage possesses no antibiotic resistance genes (ARGs), mobile genetic elements (MGEs), and any E. coli virulence associated genes. In vitro bacterial challenge tests demonstrated that two phages, the Escherichia phage VaT-2019a isolate PE17 and the Escherichia phage AG- MK-2022. Basu exhibited high bactericidal activity against APEC strains and lysed 95% of the tested APEC strains. Conclusions: The current study findings indicate that both phages could be suggested as safe biocontrol agents and alternatives to antibiotics for controlling MDR-APEC strains isolated from broilers. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas aeruginosa is anaerobic, motile gram-negative rod which is responsible for 10% of all hospital acquired or nosocomial infections. Present study was conducted to determine the incidence, risk factors and antimicrobial resistance pattern of P. aeruginosa isolated from different clinical samples. Materials and method: A total of 7293 patients presenting with different complains in different OPD and IPD of our hospital were included in the study. From all the 7293patients' clinical samples such as pus, swab, urine, sputum, blood, cerebrospinal fluid, pleural fluid, peritoneal fluid, tissue biopsies and bronchial lavage were collected and received to our laboratory without delay for further processing. All the samples were processed for aerobic culture and organisms were identified by standard protocol. The P.eruginosa was tested for antimicrobial resistance by Kirby Bauer disc diffusion method according to CLSI guidelines. Results: Out of 7293 clinical samples cultured, 3865 (52.9%) yielded significant growth and the rest 3428 (47.1%) samples were either sterile or showed non-significant growth. From 3865 growth positive samples, a total of 355 (9.18%) P. aeruginosa were isolated. From 355 isolates, majority 249 isolates (70.14%) were isolated from IPD in the hospital and rest 106 isolates (29.85%) were isolated from out-patients (community-acquired infection). 206 (5.32%) of P. aeruginosa were isolated from pus and swab samples followed by urine 126 (3.26%) and blood 01 (0.02%) samples. The maximum age of patients infected with P. aeruginosa was 31-40 years old and male to female ratio was 1.25:1. Pseudomonas aeruginosa was highly resistant to Ceftazidime 81.97%, Cefepime 64.50%, Piperacillin 41.42%, Ciprofloxacin 44.50%, Levofloxacin 36.05%, and Amikacin 30.42%. Conclusion: This type of epidemiological study will help the better infection control strategies in hospitals and improve the knowledge of antibiotic resistance patterns among clinicians so that the appropriate antibiotic prescribed to the patients and also control of irrelevant, irrational use of antibiotic. Thus, there is a need for periodical antimicrobial surveillance to monitor the resistance patterns in tertiary care hospitals. Study shows the frequency of multidrug resistant among P. aeruginosa. [ABSTRACT FROM AUTHOR]

Introduction: The colonization of the oral cavity by potentially pathogenic antimicrobial-resistant bacteria in adolescents and its consequences is very poorly understood. The present study focused on the occurrence of oral colonization by Gram-negative bacilli (GNB) and their multidrug resistance, including the production of extended-spectrum β-lactamases (ESBLs) and carbapenemases, among healthy adolescents and risk factors associated with GNB colonization. Materials and methods: This study was conducted as part of "A program for the early detection of risk factors for lifestyle diseases SOPKARD-Junior" (SOPKARDJunior). Oral samples were collected from 182 adolescents from four public elementary schools in Sopot, Poland, aged 13-14 years. Bacterial strains were identified by the MALDI-TOF MS method. Screening of antimicrobial resistance was performed using a disk diffusion method. The NG-Test® CARBA-5 was used to detect and differentiate the five most widely distributed carbapenemases. Demographic and clinical data were collected and statistical analysis of risk factors was performed. Results: A total of 68 out of 182 (37.4%) healthy adolescents was documented oral colonization with Gram-negative bacilli, including 50/182 (27.5%) multidrug resistant (MDR-GNB) strains. Over 60% of oral carriage concerned three main genera Enterobacter spp., Pseudomonas spp., and Serratia spp., which were detected in 22.1%, 19.1%, and 19.1% of participants, respectively. Citrobacter spp., Escherichia coli, Klebsiella spp., Hafnia spp., Aeromonas spp., Acinetobacter spp., and Stenotrophomonas spp. were also isolated. The antimicrobial resistance to ampicillin (100%), ceftazidime (69.1%), meropenem (60.3%), gentamycin (60.3%), piperacillin/tazobactam (52.9%), and piperacillin (45.6%) were the most common. Among 73.5% GNB strains multidrug resistance was observed, including all Pseudomonas spp. strains. Among MDR-GNB, 30.4% were resistant to four groups of antibiotics, half of the MDR Pseudomonas spp. strains were resistant to 10 groups of antibiotics. Extended-spectrum β-lactamases were produced by Enterobacter cloacae, Klebsiella spp., and Serratia spp. (7.4%). Colonization by ESBLs-positive GNB strains was significantly associated with recurrent respiratory infections, nasal congestion, and bronchitis (p<0.05). Conclusion: Our study revealed high oral carriage of multi-drug resistant Gramnegative bacilli in healthy adolescents and the association of ESBL-producing strains with respiratory infections. Further studies on oral colonization with GNB are necessary due to the possibility of distinct infections and the acquisition of antibiotic resistance by resident microbiota. [ABSTRACT FROM AUTHOR]

IntroductionThe colonization of the oral cavity by potentially pathogenic antimicrobial-resistant bacteria in adolescents and its consequences is very poorly understood. The present study focused on the occurrence of oral colonization by Gram-negative bacilli (GNB) and their multidrug resistance, including the production of extended-spectrum β-lactamases (ESBLs) and carbapenemases, among healthy adolescents and risk factors associated with GNB colonization.Materials and methodsThis study was conducted as part of “A program for the early detection of risk factors for lifestyle diseases SOPKARD-Junior” (SOPKARD-Junior). Oral samples were collected from 182 adolescents from four public elementary schools in Sopot, Poland, aged 13-14 years. Bacterial strains were identified by the MALDI-TOF MS method. Screening of antimicrobial resistance was performed using a disk diffusion method. The NG-Test® CARBA-5 was used to detect and differentiate the five most widely distributed carbapenemases. Demographic and clinical data were collected and statistical analysis of risk factors was performed.ResultsA total of 68 out of 182 (37.4%) healthy adolescents was documented oral colonization with Gram-negative bacilli, including 50/182 (27.5%) multidrug resistant (MDR-GNB) strains. Over 60% of oral carriage concerned three main genera Enterobacter spp., Pseudomonas spp., and Serratia spp., which were detected in 22.1%, 19.1%, and 19.1% of participants, respectively. Citrobacter spp., Escherichia coli, Klebsiella spp., Hafnia spp., Aeromonas spp., Acinetobacter spp., and Stenotrophomonas spp. were also isolated. The antimicrobial resistance to ampicillin (100%), ceftazidime (69.1%), meropenem (60.3%), gentamycin (60.3%), piperacillin/tazobactam (52.9%), and piperacillin (45.6%) were the most common. Among 73.5% GNB strains multidrug resistance was observed, including all Pseudomonas spp. strains. Among MDR-GNB, 30.4% were resistant to four groups of antibiotics, half of the MDR Pseudomonas spp. strains were resistant to 10 groups of antibiotics. Extended-spectrum β-lactamases were produced by Enterobacter cloacae, Klebsiella spp., and Serratia spp. (7.4%). Colonization by ESBLs-positive GNB strains was significantly associated with recurrent respiratory infections, nasal congestion, and bronchitis (p

Arwa R Elmanakhly,1 Mahmoud M Bendary,2 Nesreen A Safwat,1 Eman Abu-Elnasr Awad,3 Majid Alhomrani,4,5 Abdulhakeem S Alamri,4,5 El-Sayed Khafagy,6,7 Hadil Faris Alotaibi,8 Fatma I Abou-Elazm9 1Department of Microbiology and Immunology, Faculty of Pharmacy, Modern University for Technology and Information, Cairo, 11559, Egypt; 2Microbiology and Immunology Department, Faculty of Pharmacy, Port Said University, Port Said, 42511, Egypt; 3Department of Internal Medicine, Faculty of Medicine for Girls, Al Azhar University, Cairo, 11559, Egypt; 4Department of Clinical Laboratories Sciences, The Faculty of Applied Medical Science, Taif University, Taif, 26432, Saudi Arabia; 5Centre of Biomedical Science Research (CBSR), Deanship of Scientific Research, Taif University, Taif, 26432, Saudi Arabia; 6Department of Pharmaceutics, College of pharmacy, Prince Sattam Bin Abdulaziz University, Al-kharj, 11942, Saudi Arabia; 7Department of Pharmaceutics and Industrial Pharmacy, Suez Canal University, Ismailia, 41522, Egypt; 8Department of Pharmaceutical Science, College of Pharmacy, Princess Nourah Bint Abdulrahman University, Riyadh, 11671, Saudi Arabia; 9Department of Microbiology and Immunology, Faculty of Pharmacy, Misr University for Science and Technology, Cairo, 11559, EgyptCorrespondence: Mahmoud M Bendary, Tel +1227550629, Email m.pendary@pharm.psu.edu.egBackground: Klebsiella pneumoniae (K. pneumoniae) is one of the most important pathogens in nosocomial infections. It has resistance to most antibiotics, even carbapenem, resulting in restricted therapeutic options.Purpose: We tried to assess the antimicrobial resistance and virulence fitness of carbapenem-resistant K. pneumoniae (CRKP) in addition to their phenotypic and genotypic diversity.Materials and Methods: The conventional methods, automated Vitek-32 system, and antimicrobial susceptibility pattern were used to detect CRKP isolates. Virulence and resistance genes profiles were created by using PCR technique. The correlation analysis was done by using R-program.Results: The antimicrobial resistance profile for all our K. pneumoniae isolates was shocking as the MDR and CRKP were the most prominent phenotypes. Unfortunately, high degrees of heterogeneity among our CRKP isolates were recorded, as 97.5% of them were differentiated into different clusters. We found a negative correlation between the existence of virulence and antimicrobial resistance genes. In contrast to sputum and urine CRPK isolates, the blood isolates showed high antimicrobial resistance and low virulence fitness. Finally, K. pneumoniae creates several outbreaks and crises in Egypt owing to the highly heterogeneity and the wide spread of multidrug-resistant (MDR) and multi-virulent CRKP phenotypes.Conclusion: Our results are significant and alarming to health organizations throughout the world for the severity and heterogeneity of K. pneumoniae infections. Therefore, the traditional method for treatment of CRKP infections must be renewed. Additionally, the treatment protocols must be well correlated with the site of infections, phenotypes, and genotypes of CRKP strains.Keywords: K. pneumoniae, virulence genes, heterogeneity, multidrug resistant, MDR, carbapenem resistance K. pneumonia, CRPK

Background: Ventilator-associated pneumonia (VAP) is the most common nosocomial infection diagnosed in intensive care units (ICU) worldwide. VAP is an infection that occurs in an ICU patient more than 48 hours after endotracheal intubation and mechanical ventilation. This study aims to determine the aetiological agents causing bacterial ventilator-associated pneumonia. Materials and Methods: A retrospective descriptive study was performed on all patients with VAP that were admitted to the ICU at Sungai Buloh Hospital that fit the inclusion criteria from January 2012 to December 2014. The data used for this study was reviewed after retrieving it from the information system (EHIS) of Sungai Buloh Hospital. Result: The results showed the Acinetobacter spp. were the most common pathogens isolated (33%) and 83.9% were multi-drug resistant (MDR). A high proportion (75%) of the Staphylococcus aureus isolates were methicillin-resistant Staphylococcus aureus (MRSA). Conclusion: The finding of the study can be used to guide clinicians in the management of patients with VAP. The pitfalls in using empiric antibiotics for suspicion of VAP are the potential for antibiotic overuse, emergence of resistance, unnecessary adverse effects and potential toxicity. The major goals of VAP management are early and, appropriate antibiotics in adequate doses followed by de-escalation based on microbiological culture results and the clinical response of the patient. [ABSTRACT FROM AUTHOR]

• Escherichia coli were isolated from 90 children under the age of 5 years presenting with diarrhoea at clinics and hospitals in Limpopo province, South Africa. • 98.8 % (164/166) of isolates were categorized as multidrug resistant. • 4.9 % (9/166) of isolates were categorized as extensively drug resistant. • 92.2 % (153/166) of isolates were phenotypically extended-spectrum β-lactamase producers. Globally, millions of children die as a result of diarrhoea and/or antimicrobial resistant infections. Diarrhoeagenic Escherichia coli (DEC) are responsible for a substantial proportion of cases of diarrhoea in South Africa and sub-Saharan Africa. Effective treatments (including the use of antimicrobials) are therefore essential. E. coli isolated from children under the age of five were subjected to antimicrobial susceptibility testing using the Vitek 2® compact automated system (bioMérieux Inc., France) and categorized as multidrug or extensively drug resistant (MDR or XDR). Almost all isolates (164/166, 98.8 %) were categorized as MDR with 4.9 % (9/166) categorized as XDR. The majority of isolates (153/166, 92.2 %) were also phenotypically classified as extended-spectrum β-lactamase (ESBL) producers. More than half of these isolates (78/153, 51.0 %) were subjected to PCR for genes associated with ESBL production. More than half (45/78, 57.7 %) of the isolates tested were PCR positive for at least one ESBL gene or gene group and 11.5 % (9/78) were positive for two ESBL genes or gene groups. There is a need to strengthen antimicrobial resistance surveillance in South Africa and improve infection prevention and control measures. There is also a need to review the current South African Treatment Guidelines as outlined by the South African Essential Drugs Programme. [Display omitted] [ABSTRACT FROM AUTHOR]

Objectives: The objective of this review was to compare the effectiveness of Colistin monotherapy and combination therapy for the treatment of multidrug-resistant gram-negative bacterial infections. Data sources: PubMed, Cochrane Library. Study eligibility, interventions, and exclusions: In this systematic review, we included all retrospective and prospective studies and randomized controlled trials (RCTs) that compared intravenous polymyxin monotherapy and combination therapy with any other antibiotic for treating multidrug-resistant infections. Studies using inhaled polymyxins with 5 or less than 5 patients were excluded. The primary outcome was 30-day all-cause mortality and if not reported at day 30 we extracted and documented the closest time point. Both crude outcome rates and adjusted effect estimates were extracted for mortality. Study appraisal, data extraction and synthesis: Search string used was "(Colistin OR polymyxin) AND (Enterobacteriaceae OR Klebsiella OR Acinetobacter OR Escherichia coli OR Pseudomonas) AND (random OR prospective OR retrospective OR cohort OR observational OR blind)." Thirty-nine studies were included in our analysis; out of which 6 RCTs were included and 9 studies used carbapenem as the adjunctive antibiotic. Each study was screened and reviewed for eligibility independently by two authors and data extrapolated on an Excel sheet. Results: The meta-analysis of polymyxin monotherapy vs. combination therapy in multidrug-resistant infections yielded an odds ratio (OR) of 0.81 (95% confidence interval [CI]: 0.65-1.01) with minimal heterogeneity (I² = 40%), whereas pooled analysis of this comparison in studies that included carbapenem as combination therapy yielded an OR of 0.64 (CI: 0.40-1.03; I² = 62%). Likewise, the pooled analysis of the RCTs yielded an OR of 0.82 (95% CI: 0.58-1.16, I² = 22%). All these showed no statistical significance. However, it was seen that polymyxin combination therapy was more effective in multidrug-resistant infections compared to polymyxin monotherapy. The effectiveness was more glaring when carbapenems were used as the combination drug instead of any other antibiotic and more so in many in vitro studies that used polymyxin combination therapy. Conclusion: Although statistically insignificant, it would be prudent to use polymyxin combination therapy to treat multidrug-resistant gram-negative bacilli (GNB) infection over monotherapy with preference to use carbapenem as the adjunct alongside polymyxins. [ABSTRACT FROM AUTHOR]

Introduction Antimicrobial resistance (AMR) has become a menace, spreading among bacterial species globally. AMR is now recognized as a silent pandemic responsible for treatment failures. Therefore, an effective surveillance mechanism is warranted to understand the bacterial species isolated from human clinical specimens. The present study employed next-generation sequencing (NGS) or whole-genome sequencing (WGS) to identify the resistance and virulence genes, sequence type, and serotypes. Methods This study included 18 multidrug-resistant (MDR) Klebsiella pneumoniae ( K . pneumoniae ) isolates obtained from patients suffering from different infections attending the Prathima Institute of Medical Sciences, Karimnagar, India. All isolates were identified, and antimicrobial susceptibility profiles were determined through conventional microbiological techniques and confirmed by automated systems. All the isolates were investigated using NGS or WGS to identify the genes coding for resistance, such as extended-spectrum beta-lactamases (ESBLs), metallo-beta-lactamases, and virulence genes. Multilocus sequence typing (MLST) was conducted to identify the sequence types, and Kleborate analysis was performed to confirm the species, genes for AMR, and virulence and evaluate the capsular polysaccharide (KL) and cell wall/lipopolysaccharide (O) serotypes carried by the isolates. Results The mean age of the patients was 46.11±20.35 years. Among the patients included, 12 (66.66%) were males and 6 (33.33%) were females. A high percentage (>50%) of hypervirulent K . pneumoniae (hvKp) strains that had genes coding for AMR and plasmids having the potential to carry  bla NDM  and resistance genes were observed. Among the isolates, 16 (88.88%) revealed the presence of multiple antibiotic-resistant genes with evidence of at least one gene coding for beta-lactamase resistance. There was a high prevalence of bla SHV (17/18; 94.44%) and bla CTX-M-15 (16/18; 88.88%) AMR genes. Other AMR genes identified included bla TEM (83.33%; 15/18) and bla OXA (14/18; 77.77%). Two (11.11%) strains each showed the presence of bla NDM-1 and bla NDM-5 genes. The virulence genes identified included  gapA , infB , mdh , pgi , phoE , rpoB , tonB , and ybt . The most frequent K . pneumoniae serotypes found were KL51:O1v2 (3/18, 16.66%), KL17:O1v1 (3/18, 16.66%), and KL64:O2v1 (3/18, 16.66%). KL64 (4/18; 22.22%) was the most common capsular serotype identified among the isolates. The most frequent MLST-based sequence type (ST) identified included ST-147 (5/18, 27.77%), followed by ST-231 (3/18, 16.66%) and ST-101 (2/18, 11.11%). Conclusions The molecular analysis of  K . pneumoniae  isolates revealed multiple AMR, plasmid, and virulence genes. Additionally, many global STs were noticed by MLST. The results noted a high prevalence of hvKp strains. Molecular characterization of bacterial strains using NGS/WGS is important to understand the epidemiology of bacterial strains and the antibiotic resistance and virulence genes they are potentially carrying. The data obtained from this study may be utilized to devise careful antibiotic-prescribing approaches and improve patient management practices., Competing Interests: Kempegowda Institute of Medical Sciences has made a memorandum of understanding (MOU) with Dr. Venkataramana Kandi, Prathima Institute of Medical Sciences, to collaborate under the project supported by the Global Health Research Unit (GHRU), United Kingdom, (Copyright © 2024, Moses et al.)

Every year millions of people die due to fatal waterborne diseases around the world especially in developing countries like India. Sikkim, a northeastern state of India, greatly depends on natural water sources. About 80% of the population of Sikkim depends on natural spring water for domestic as well as agricultural use. Recent waterborne disease outbreaks in the state raises a concerning question on water quality. In this study, we analyzed water quality especially for the detection of Enterobacteriaceae members from four districts of the state. Isolation with selective culture media techniques and taxonomic characterization of Enterobacteriaceae bacteria with 16S rRNA gene showed the prevalence of Escherichia coli (37.50%), Escherichia fergusonii (29.41%), Klebsiella oxytoca (36.93%), Citrobacter freundii (37.92%), Citrobacter amalonaticus (43.82%), Enterobacter sp. (43.82%), Morganella morganii (43.82%), Hafnia alvei (32.42%), Hafnia paralvei (38.74%), and Shigella flexneri (30.47%) in the spring water of Sikkim. Antibiotic susceptibility test (AST) showed resistance of the isolates to common antibiotics like ampicillin, amoxicillin as well as to third generation antibiotics like ceftazidime and carbapenem. None of the isolates showed resistance to chloramphenicol. E. coli isolated from spring water of Sikkim showed presence of different virulence genes such as stx1 (81.81%), elt (86.66%), and eae (66.66%) along with resistance gene for ampicillin (CITM) (80%), quinolones (qnrB) (44.44%), tetracycline (tetO) (66.66%), and streptomycin (aadA1) (66.66%). The data indicates a high incidence rate of multiple antibiotic resistant enteric bacteria in the spring water of Sikkim. Additionally, the presence of enteric bacteria in the water samples indicates widespread fecal contamination of the spring water.

Pseudomonas aeruginosa is a Gram-negative bacterium which is capable of developing a high level of antibiotic resistance. It has been placed on the WHO’s critical priority pathogen list and it is commonly found in ventilator-associated pneumonia infections, blood stream infections and other largely hospital-acquired illnesses. These infections are difficult to effectively treat due to their increasing antibiotic resistance and as such patients are often treated with antibiotic combination regimens. Methods: We conducted a systematic search with screening criteria using the Ovid search engine and the Embase, Ovid Medline, and APA PsycInfo databases. Results: It was found that in many cases the combination therapies were able to match or outperform the monotherapies and none performed noticeably worse than the monotherapies. However, the clinical studies were mostly small, only a few were prospective randomized clinical trials and statistical significance was lacking. Conclusions: It was concluded that combination therapies have a place in the treatment of these highly resistant bacteria and, in some cases, there is some evidence to suggest that they provide a more effective treatment than monotherapies.

Every year millions of people die due to fatal waterborne diseases around the world especially in developing countries like India. Sikkim, a northeastern state of India, greatly depends on natural water sources. About 80% of the population of Sikkim depends on natural spring water for domestic as well as agricultural use. Recent waterborne disease outbreaks in the state raises a concerning question on water quality. In this study, we analyzed water quality especially for the detection of Enterobacteriaceae members from four districts of the state. Isolation with selective culture media techniques and taxonomic characterization of Enterobacteriaceae bacteria with 16S rRNA gene showed the prevalence of Escherichia coli (37.50%), Escherichia fergusonii (29.41%), Klebsiella oxytoca (36.93%), Citrobacter freundii (37.92%), Citrobacter amalonaticus (43.82%), Enterobacter sp. (43.82%), Morganella morganii (43.82%), Hafnia alvei (32.42%), Hafnia paralvei (38.74%), and Shigella flexneri (30.47%) in the spring water of Sikkim. Antibiotic susceptibility test (AST) showed resistance of the isolates to common antibiotics like ampicillin, amoxicillin as well as to third generation antibiotics like ceftazidime and carbapenem. None of the isolates showed resistance to chloramphenicol. E. coli isolated from spring water of Sikkim showed presence of different virulence genes such as stx1 (81.81%), elt (86.66%), and eae (66.66%) along with resistance gene for ampicillin (CITM) (80%), quinolones (qnrB) (44.44%), tetracycline (tetO) (66.66%), and streptomycin (aadA1) (66.66%). The data indicates a high incidence rate of multiple antibiotic resistant enteric bacteria in the spring water of Sikkim. Additionally, the presence of enteric bacteria in the water samples indicates widespread fecal contamination of the spring water. [ABSTRACT FROM AUTHOR]

Due to their propensity for causing diarrheal illnesses and their rising susceptibility to antimicrobials, Shigella infections constitute a serious threat to global public health. This extensive study explores the frequency, antibiotic resistance, genetic evolution, and effects of Shigella infections on vulnerable groups. The research covers a wide range of geographical areas and sheds information on how the prevalence of Shigella species is evolving. Shigella strain antimicrobial resistance patterns are thoroughly examined. Multidrug resistance (MDR) has been found to often occur in investigations, especially when older antimicrobials are used. The improper use of antibiotics in China is blamed for the quick emergence of resistance, and variations in resistance rates have been seen across different geographical areas. Shigella strains' genetic makeup can be used to identify emerging trends and horizontal gene transfer's acquisition of resistance genes. Notably, S. sonnei exhibits the capacity to obtain resistance genes from nearby bacteria, increasing its capacity for infection. The study also emphasizes the difficulties in accurately serotyping Shigella strains due to inconsistencies between molecular and conventional serology. These results highlight the necessity of reliable diagnostic methods for monitoring Shigella infections. In conclusion, this study emphasizes how dynamic Shigella infections are, with varying patterns of occurrence, changing resistance landscapes, and genetic adaptability. In addition to tackling the rising problem of antibiotic resistance in Shigella infections, these findings are essential for guiding efforts for disease surveillance, prevention, and treatment., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2024, Satija et al.)

The aim of this study is to see the frequency, clinical presentation, and therapeutic response of extensively drug-resistant Salmonella enterica serovar Typhi and current susceptibility pattern of typhoidal Salmonella strains in our setup. This study was carried out at the Department of Medical Microbiology and Immunology and Department of Medicine, Pakistan Navy Ship (PNS) Shifa Hospital, Karachi, from January 1 to December 31, 2018. All the blood culture samples of patients (indoor and outdoor) with suspicion of enteric fever were processed. Isolates were cultured and identified using standard microbiological procedures. The antimicrobial sensitivity against the typhoidal Salmonellae was determined using Kirby-Bauer disc diffusion method as per the guidelines of Clinical and Laboratory Standards Institute (2018) and all the extensively drug-resistant (XDR) isolates were confirmed by Vitek 2 system. Clinical presentation and response to treatment of patients were followed. A total of 292 typhoidal Salmonella isolates were cultured. Resistance to ciprofloxacin against both Salmonella Typhi and Salmonella Paratyphi A was found to be very high (91%). Percentage of multidrug-resistant (MDR) isolates in Salmonella Typhi was 76% (182 isolates) and in Salmonella Paratyphi it was 34% (18 isolates). XDR isolates in Salmonella Typhi were significant that is 48% (115 isolates). Only 10 cases were given azithromycin who responded to treatment in mean 4.3 days. Out of 115 cases of XDR Salmonella Typhi, 103 patients were given parenteral meropenem and clinical response was seen in mean 5 days. The emergence and rapid spread of extensively drug-resistant Salmonella Typhi is alarming and highlights the significance of strict antimicrobial susceptibility surveillance programs with antimicrobial stewardship. [ABSTRACT FROM AUTHOR]

The antibiotic resistance is creating a serious challenge to treat diseases especially against multidrug resistant (MDR) E. coli. The present study was conducted to evaluate the antibacterial activity of seven different flowers ethanolic extracts named as Achillea millefolium, Bombax ceiba, Chrysanthemum cinerarifolium, Hyssopus officinalis, Rosa damascena Miller, Taraxacum officinale Weber and Woodfordia fruticosa against MDR E. coli through spot test and minimum inhibitory concentrations (MIC) test. The results showed all the seven extracts have significant antibacterial activity against MDR E. coli and control. The MIC of Rosa damascena Miller, Bombax ceiba was 3.125 mg/ml and Taraxacum officinale Weber was 12.5 mg/ml for MDR E. coli. Similarly, Achillea millefolium, Hyssopus officinalis, Chrysanthemum cinerarifolium MIC value was 25 mg/ml and 12.5 mg/ml for Woodfordia fruticosa Kurtz. The current study reveals that these flowers possess strong antibacterial activity and thus can be used as potential antimicrobial agent against various resistant bacterial pathogens. [ABSTRACT FROM AUTHOR]

Mycobacterium abscessus complex consist of three rapidly growing subspecies: M. abscessus, M. massiliense, and M. bolletii. They are clinically important human pathogens responsible for opportunistic pulmonary and skin and soft tissue infections. Treatment of M. abscessus infections is difficult due to in vitro resistance to most antimicrobial agents. Tedizolid (TZD) is a next-generation oxazolidinone antimicrobial with a wide spectrum of activity even against multidrug resistant Gram-positive bacteria. In this study, the in vitro activity of TZD against the M. abscessus complex (n = 130) was investigated. Susceptibility testing by broth microdilution showed lower TZD minimum inhibitory concentrations (MICs) when compared to linezolid. The MIC50 and MIC90 was 1 mg/L and 4 mg/L, respectively across all M. abscessus complex members, reflecting no difference in subspecies response to TZD. Pre-exposure of M. abscessus complex to subinhibitory concentrations of TZD did not trigger any inducible drug resistance. Single-drug time kill assays and bactericidal activity assays demonstrated bacteriostatic activity of TZD in all three M. abscessus subspecies, even at high drug concentrations of 4 to 8x MIC. Combination testing of TZD with clarithromycin, doxycycline and amikacin using the checkerboard approach showed no antagonistic interactions. TZD may be an effective therapeutic antimicrobial agent for the treatment of M. abscessus infections.

Mycobacterium abscessus complex consist of three rapidly growing subspecies: M. abscessus, M. massiliense , and M. bolletii. They are clinically important human pathogens responsible for opportunistic pulmonary and skin and soft tissue infections. Treatment of M. abscessus infections is difficult due to in vitro resistance to most antimicrobial agents. Tedizolid (TZD) is a next-generation oxazolidinone antimicrobial with a wide spectrum of activity even against multidrug resistant Gram-positive bacteria. In this study, the in vitro activity of TZD against the M. abscessus complex (n = 130) was investigated. Susceptibility testing by broth microdilution showed lower TZD minimum inhibitory concentrations (MICs) when compared to linezolid. The MIC50 and MIC90 was 1 mg/L and 4 mg/L, respectively across all M. abscessus complex members, reflecting no difference in subspecies response to TZD. Pre-exposure of M. abscessus complex to subinhibitory concentrations of TZD did not trigger any inducible drug resistance. Single-drug time kill assays and bactericidal activity assays demonstrated bacteriostatic activity of TZD in all three M. abscessus subspecies, even at high drug concentrations of 4 to 8x MIC. Combination testing of TZD with clarithromycin, doxycycline and amikacin using the checkerboard approach showed no antagonistic interactions. TZD may be an effective therapeutic antimicrobial agent for the treatment of M. abscessus infections. [ABSTRACT FROM AUTHOR]

A total of 189 Salmonella isolates were recovered from 627 samples which were collected from cecal contents of broilers, chicken carcasses, chicken meat after cutting step and frozen broiler chicken products along the slaughtering process at a slaughterhouse in Sichuan province of China. The Salmonella isolates were subjected to antimicrobial susceptibility testing to 10 categories of antimicrobial agents using the Kirby–Bauer disk diffusion method. Those antibiotics-resistant isolates were further investigated for the occurrence of resistance genes, the presence of class 1 integron as well as the associated gene cassettes, and the mutations within the gyrA and parC genes. Consequently, the prevalence of Salmonella was 30.14% (47.96% for cecal content, 18.78% for chicken carcasses, 31.33% for cutting meat and 14.00% for frozen meat, respectively). The predominant serotypes were S. Typhimurium (15.34%) and S . Enteritidis (69.84%). High resistance rates to the following drugs were observed: nalidixic acid (99.5%), ampicillin (87.8%), tetracycline (51.9%), ciprofloxacin (48.7%), trimethoprim/sulfamethoxazole (48.1%), and spectinomycin (34.4%). Antimicrobial resistance profiling showed that 60.8% of isolates were multidrug resistant (MDR), and MDR strains increased from 44.7% to 78.6% along the slaughtering line. 94.6% ( n = 157) of beta-lactam-resistant isolates harbored at least one resistance gene of bla TEM or bla CTX-M . The relatively low prevalence of aminoglycoside resistance genes ( aac(3)-II , aac(3)-IV , and ant(2″)-I ) was found in 49 (66.2%) of antibiotic-resistant isolates. The tetracycline resistance genes ( tet (A), tet (B), tet (C), and tet (G) and sulfonamide resistance genes ( sul1 , sul2 , and sul3 ) were identified in 84 (85.7%) and 89 (97.8%) antibiotic-resistant isolates respectively. floR was identified in 44 (97.8%) florfenicol-resistant isolates. Class 1 integron was detected in 37.4% ( n = 43) of the MDR isolates. Two different gene cassettes, bla OXA-30 - aadA1 (19 isolates) and bla OXA-30 -aadA1 / drfA1 - orfC (2 isolates), were identified in class 1 integron-positive isolates. 97.9% (184/188) of quinolone-resistant isolates had at least one mutation within gyrA or parC . Overall, antimicrobial resistance showed an increasing trend along the slaughtering process. The results showed that broiler chicken products in the slaughterhouse were contaminated with MDR Salmonella , which might originate from food producing animals to some extent, and cross-contamination during slaughter, and facilitate the dissemination of the resistance genes to consumers along the production chain, which suggests importance of controlling Salmonella during slaughter for public health, underlying strict hygiene method and HACCP management to reduce cross-contamination. [ABSTRACT FROM AUTHOR]

Background and Objective: Typhoid fever caused by Salmonella typhi (S. typhi) is an important cause of morbidity and mortality worldwide. The emergence of resistance against the first and second line of drugs has limited the range of drug choices for its treatment. Therefore, the present study was designed to assess the status of drug resistance in S. typhi isolated from the blood samples of pediatric patients reporting at a leading tertiary care hospital in Lahore. Methods: This descriptive study was conducted at the Pathology Department of Allama Iqbal Medical College, Lahore, Pakistan, over a period of 2 years. A total of 1,306 blood samples were aseptically collected from the pediatric population of Jinnah Hospital, Lahore, Pakistan, after obtaining informed assent from the parents/guardians. The samples were transferred into a Bact/Alert blood culture bottle. After the detection of microbial growth by the Bact/Alert 3-D system, bacterial isolates were identified by standard microbiological procedures. VITEK 2 compact automated identification and antimicrobial susceptibility testing instrument (bioMerieux) was used for the antimicrobial sensitivity testing. Data were entered and analyzed by using Statistical Package for the Social Sciences version 21.0. Results: A total of 235 out of 1,306 (17.9%) blood culture samples yielded bacterial growth, among which 62.5% (n = 147) were S. typhi. Among 147 S. typhi isolates, 70.6% were multidrug resistant, while 63% were extensively drug resistant (XDR). Maximum resistance against first-line drugs observed was for ampicillin (79.4) and co-trimoxazole (67.2). Conclusion: The emergence and spread of XDR S. typhi with a high level of resistance are quite alarming, leaving limited treatment options for the pediatric population. Moreover, increasing resistance to antibiotics demonstrates the quick waning of the efficacy of available first and second-line drugs in the treatment of typhoid fever. [ABSTRACT FROM AUTHOR]

Bioassay guided fractionation of an antimycobacterial extract of Foeniculum vulgare var dulce (Apiaceae) led to the isolation and characterization of 5-hydroxyfurano-coumarin. The chemical structure of this compound was elucidated by 1H and 13C (1D and 2D) Nuclear Magnetic Resonance (NMR) spectroscopy. In addition, the active fractions were analyzed by GC-MS and seventy eight compounds were identified; the major compounds were 1,3-benzenediol, 1-methoxycyclohexene, o-cymene, sorbic acid, 2-hydroxy-3-methyl-2-cyclopenten-1-one, estragole, limonene-10-ol and 3-methyl-2-cyclopenten-1-one. Twenty compounds identified in the active fractions were tested against one sensitive and three MDR strains of Mycobacterium tuberculosis using the Alamar Blue microassay. Compounds that showed some degree of antimycobacterial activity against all strains tested were the following: linoleic acid (MIC 100 µg/mL), oleic acid (MIC 100 µg/mL), 1,3-benzenediol (MIC 100–200 µg/mL), undecanal (MIC 50–200 µg/mL), and 2,4-undecadienal (MIC 25–50 µg/mL), the last being the most active compound. To our knowledge, this is the first report of the presence of 5-hydroxy-furanocoumarin in F. vulgare.

Background Following the pandemic caused by SARS-CoV-2, the emergence of and following the pandemic has required major modifications to healthcare systems and frameworks. Antimicrobials have more than a few potential roles in managing COVID-19. Experimental cures for the treatment of SARS-CoV-2 are being explored. The availability of limited data suggests that nosocomial infections are associated with a higher risk of death and severity of COVID-19. To fill this knowledge gap, we conducted a study to assess the spectrum of bacteriological isolates in different samples from COVID-19 patients. Our study aimed to evaluate the antibiotic resistance pattern of these bacterial isolates and compare the spectrum of bacteriological isolates in different samples and their antibiotic sensitivity pattern between COVID-19 and non-COVID-19 patients. Methodology An observational cross-sectional, partly retrospective, and partly prospective study was carried out in the bacteriology section of the Department of Microbiology, Indira Gandhi Institute of Medical Sciences (IGIMS), Patna, Bihar, for a total duration of six months from February 2021 to July 2021. The profile of pathogens isolated from 105 clinical samples from COVID-19 patients was studied. To detect COVID-19, RT-PCR was performed. All clinical specimens (urine, blood, pus, respiratory sample, etc.) were processed and cultured on different media to support the growth of the bacteria as per our standard operating procedures (SOP) for bacteriology samples. Antimicrobial susceptibility testing was carried out based on the Clinical and Laboratory Standards Institute. Results A total of 105 clinical samples were received in the bacteriology section of the Department of Microbiology, IGIMS, Patna, Bihar, from admitted COVID-19 patients. The mean age of study participants was 51.57 ± 14.76 years, and males (66.7%, 70/105) were more than females (33.3%, 35/105). The majority of the patients were 91 out of 105 (86.67%) from the ward and 14 from the ICU (13.33%). Of the total samples tested, 62 (59%) were urine samples, 26 (24.8%) were respiratory specimens, 13 (12.4%) were pus samples, 3 (2.9%) were body fluids, and 1 (1%) were tissue samples. Among the total pathogen isolates (n=57) obtained from patients with SARS-CoV-2 admitted to the ward and ICU, 56.14% (32) were gram-negative, 26.31% (15) were Candida , and 17.54% (10) were gram-positive pathogens. The most isolated pathogen was Escherichia coli (39.02%, 16/41) followed by Klebsiella pneumoniae (29.26%, 12/41), Acinetobacter baumannii (7.31%, 3/41), and Enterobacter cloacae (2.43%, 1/41). Enterococcus spp. as gram-positive bacteria were isolated in 21.95% (9/41) of patients. Among the gram-negative bacteria ( Enterobacterales ), the highest resistance was seen in ampicillin (100%,29/29). For non- Enterobacterales , the highest resistance was seen in ceftriaxone (66.66%,2/3). Enterococcus spp. showed maximum ciprofloxacin, and gentamicin (high level) resistance was 100% (9/9). Conclusion Secondary infections with resistant pathogens in COVID-19 patients highlight the importance of antimicrobial management programs focused on the optimal selection of empirical treatments based on culture reports., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Arun et al.)

Objective: B. contaminans was cultured from respiratory secretions and liquid docusate (Colace) in a Neurosurgical Intensive Care Unit (NICU) patient with community-acquired Legionnaire's disease but not from another bottle given to the patient. Unexpectedly, C. pelliculosa was cultured from two bottles, but not the B. contaminans bottle or respiratory secretions. Methods: B. cepacia, later identified as B. contaminans, was cultured from a bottle of liquid docusate (Colace) dispensed to a non-cystic fibrosis patient. His respiratory secretions were colonized with B. contaminans. Results: Eradication of B. contaminans colonization in the patient's respiratory secretions was attempted. With levofloxacin, B. contaminans developed multidrug resistance (MDR). Subsequent TMP-SMX therapy did not result in further MDR. Nine other ICU patients were given docusate from the same lot, but there were no other B. contaminans isolates. Conclusion: B. contaminans colonization of respiratory secretion may be difficult to eliminate. The significance of C. pelliculosa cultured from liquid docusate (Colace) remains to be elucidated. In this case, it appeared that B. contaminans may have inhibited the growth of C. pelliculosa in the same bottle. Others should be alerted to the possibility that C. pelliculosa may be present in B. contaminans-contaminated lots of liquid docusate (Colace). [ABSTRACT FROM AUTHOR]

The increasing incidence of infections caused by multidrug-resistant (MDR) or extremely drug-resistant (XDR) gram-negative organisms has led to the reemergence of colistin use. Clinical and demographic data were collected on 94 pediatric patients diagnosed with MDR or XDR gram-negative infections and treated with either a colistin-containing regimen (colistin group) or at least one antimicrobial agent other than colistin (noncolistin group). The overall clinical response rates were 65.8% in the colistin group and 70.0% in the noncolistin group ( P = 0.33). The infection-related mortality rates were 11% in the colistin group and 13.3% in the noncolistin group ( P = 0.74). There was no statistically significant difference in nephrotoxicity in the colistin and noncolistin groups. Colistin therapy was at least as effective and as safe as beta-lactam antibiotics or quinolones, with or without aminoglycosides, in the treatment of infections caused by gram-negative organisms and may be a therapeutic option in children. [ABSTRACT FROM AUTHOR]

Introduction Klebsiella pneumonia is one of the most prevalent bacteria that cause nosocomial infections, particularly in critically ill patients in the intensive care unit (ICU). Multi-drug-resistant Klebsiella pneumoniae (MDRKP) has become an urgent risk to public health as its prevalence has sharply surged around the globe in recent decades. Therefore, this research was conducted to evaluate shifts over a four-year period in drug susceptibility patterns among Klebsiella pneumoniae isolates from mechanically ventilated intensive care unit patients. Materials and methods This is a retrospective observational study conducted in a tertiary care multi-specialty hospital and teaching institute in North India and was approved by the institutional ethics committee. The research comprised Klebsiella pneumoniae isolates from endotracheal aspirates (ETA) of patients on mechanical ventilation admitted to the general intensive care unit (ICU) of our tertiary care facility. The data from January to June 2018 and January to June 2022 were collected. According to the antimicrobial resistance profile of the strains, they were categorized as susceptible, resistant to one or two antimicrobial categories, multidrug-resistant (MDR), extensively drug-resistant (XDR), or pan-drug-resistant (PDR). The criteria for MDR, XDR, and PDR were proposed by the European Centre for Disease Prevention and Control (ECDC). IBM Statistical Package for the Social Sciences (SPSS) for Windows, Version 24.0, Armonk, NY, IBM Corp., was used for data input and analysis. Results A total of 82 cases of Klebsiella pneumonia were included in the study. Of these 82 isolates, 40 were isolated over a period of six months from January to June 2018, and the remaining 42 were isolated from January to June 2022. Among the 2018 group, five strains (12.5%) were classified as susceptible, three (7.5%) as resistant, seven (17.5%) as MDR, and 25 (62.5%) as XDR. The highest percentages of antimicrobial resistance in the 2018 group were observed with amoxicillin/clavulanic acid (90%), ciprofloxacin (100%), piperacillin/tazobactam (92.5%), and cefoperazone/sulbactam (95%). In comparison, the 2022 group showed no strain as susceptible; nine strains (21.4%) were classified as resistant; three strains (7%) as MDR; and 30 strains (93%) were classified as XDR. There was a significant increase in resistance to amoxicillin, from 10% in 2018 to nil in 2022. Overall, the rate of resistant Klebsiella pneumonia (K. pneumonia) increased from 7.5% (3/40) in 2018 to 21.4% (9/42) in 2022, while XDR Klebsiella pneumonia among the mechanically ventilated ICU patients significantly increased from 62.5% (25/40) in 2018 to 71% (30/42) in 2022. Conclusion  K. pneumoniae antibiotic resistance is a real threat in Asia and requires close monitoring to be controlled. More careful attempts should be made to create a new generation of antimicrobials since the prevalence of resistance to existing medications is rising. Antibiotic resistance should be monitored and reported by healthcare institutions regularly., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Sharma et al.)

Introduction: Multiple antibiotic resistances among common bacterial pathogens have been established as an alarming public health problem elsewhere. This study was aimed to explore prevalence of Extended-spectrum beta-lactamase (ESBL) producers among multi-drug resistant (MDR) and plasmid profiling patterns of Escherichia coli isolated in Kathmandu Model hospital, Nepal over three months period. Methods: Altogether 49 isolates were randomly selected among the reported MDR E. coli and retested for their susceptibility pattern and ESBL production by Kirby-Bauer disc diffusion test. Minimum Inhibitory Concentration of the isolates towards (fluro) quinolones was determined by agar dilution method. Plasmids from isolates were extracted by alkaline lysis method. Results: All isolates were completely resistant to Amoxycillin but sensitive to Imipenem. A very high prevalence (44, 89.8%) of ESBL producers was detected. Most of the isolates were resistant to commonly used antibiotics such as Quinolones, Tetracyclines and Cotrimoxazole. Among Quinolones, the Minimum Inhibitory Concentration (MIC) range for Nalidixic acid, Ciprofloxacin and Ofloxacin were 512 - >4093, 8 - 4096 and 1 - 256 μg/ml respectively. All 31 (63.3%) plasmid-harboring isolates contained a >33.5 kb sized plasmid. Among them, seven isolates possessed multiple (2 - 7) plasmids. Overall, twelve different resistance patterns were observed among the bacteria. Based on the patterns, the high molecular weight plasmid seemed to contain most of the resistance genes. Conclusion: It is suspected that multi drug resistance and ESBL production in E. coli with resistance to Quinolones may be due to their high molecular weight plasmids. So, continuous antibiotic susceptibility test and surveillance of the plasmid and chromosome of E. coli is essential as plasmid analysis has been applied to determine the evolution and spread of antibiotic resistance among isolates. [ABSTRACT FROM AUTHOR]

Abstract: Retinoblastoma (RB) is a common malignant intraocular tumor primarily affecting children. Multidrug resistance (MDR) proteins (P-gp and MRPs) mediated chemoresistance have been considered as a major cause of treatment failure in treatment of RB. Ocular cells have shown good tolerability against moxifloxacin (MFX). Hence, the aim of present study was to investigate the effect of moxifloxacin on the functionality of MDR proteins. Furthermore, we have also examined an interaction of MFX with anticancer agents (Topotecan, etoposide and vinblastine) for RB treatment. For interaction of MFX with efflux transporter, model cell lines transfected with the efflux transporters (MDCK-MDR1 and MDCK-MRP2) were used to perform uptake and bi-directional transport experiments. Modulation of anticancer induced cell cytotoxicity, pro-inflammatory cytokines (IL-6 and IL-8) release and caspase-3 enzyme activity in presence of MFX was also evaluated. Result indicates that MFX is a substrate of both MDR1 and MRP2 efflux transporters. Furthermore elevation of anticancer uptake and bi-directional transport, reduction in IC50 cytotoxic value and modulation of antiproliferative and cytokines release in presence of MFX by anticancer agents was observed. Our results demonstrate that MFX may not only modulate the permeability of anticancer agents at efflux sites but it may also potentiate antiproliferative activity of anticancer agents in retinoblastoma cells. This study may be further extended to explore in vivo outcome of this finding. [Copyright &y& Elsevier]

Abstract: Use of nanoparticles is among the most promising strategies to overcome microbial drug resistance. This review article consists of three parts. The first part discusses the epidemiology of microbial drug resistance. The second part describes mechanisms of drug resistance used by microbes. The third part explains how nanoparticles can overcome this resistance, including the following: Nitric oxide-releasing nanoparticles (NO NPs), chitosan-containing nanoparticles (chitosan NPs), and metal-containing nanoparticles all use multiple mechanisms simultaneously to combat microbes, thereby making development of resistance to these nanoparticles unlikely. Packaging multiple antimicrobial agents within the same nanoparticle also makes development of resistance unlikely. Nanoparticles can overcome existing drug resistance mechanisms, including decreased uptake and increased efflux of drug from the microbial cell, biofilm formation, and intracellular bacteria. Finally, nanoparticles can target antimicrobial agents to the site of infection, so that higher doses of drug are given at the infected site, thereby overcoming resistance. [Copyright &y& Elsevier]

Abstract: Ethnopharmacological relevance.: Echinops giganteus, Imperata cylindrica, Piper capense and Xylopia aethiopica are four medicinal spices used in Cameroon to treat cancers. Aim of the study: The above plants previously displayed cytotoxicty against leukemia CCRF-CEM and CEM/ADR5000 cell lines as well as human pancreatic MiaPaCa-2 cells. The present study aims at emphasizing the study of the cytotoxicity and the modes of action of the above plants on a panel of ten cancer cell lines including various sensitive and drug-resistant phenotypes. The study has been extended to the isolation of the bioactive constituents from Echinops giganteus. Materials and methods: The cytotoxicity of the extracts was determined using a resazurin reduction assay, whereas the caspase-Glo assay was used to detect the activation of caspases 3/7, caspase 8 and caspase 9 in cells treated with the four extracts. Flow cytometry was used for cell cycle analysis and detection of apoptotic cells, analysis of mitochondrial membrane potential (MMP) as well as measurement of reactive oxygen species (ROS). Results: The four tested extracts inhibited the proliferation of all tested cancer cell lines including sensitive and drug-resistant phenotypes. Collateral sensitivity of cancer cells to the extract of Echinops giganteus was generally better than to doxorubicin. The recorded IC50 ranges were 3.29µg/mL [against human knockout clones HCT116 (p53 −/− ) colon cancer cells] to 14.32µg/mL (against human liver hepatocellular carcinoma HepG2 cells) for the crude extract from Echinops giganteus, 4.17µg/mL (against breast cancer cells transduced with control vector MDA-MB231 cells) to 19.45µg/mL (against MDA-MB-231 BCRP cells) for that of Piper capense, 4.11µg/mL (against leukemia CCRF-CEM cells) to 30.60µg/mL (against leukemia HL60AR cells) for Xylopia aethiopica, 3.28µg/mL [against HCT116 (p53 −/− ) cells] to 33.43µg/mL (against HepG2 cells) for Imperata cylindica and 0.11µg/mL (against CCRF-CEM cells) to 132.47µg/mL (against HL60AR cells) for doxorubicin. The four tested extracts induced apoptosis in CCRF-CEM cells via the alteration loss of MMP whilst that of Piper capense also enhanced the production of ROS. Conclusion: The studied plants are potential cytotoxic drugs that deserve more detailed exploration in the future, to develop novel anticancer drugs against sensitive and otherwise drug-resistant phenotypes. [Copyright &y& Elsevier]

Abstract: A rapid ELISA employing intact Pseudomonas aeruginosa (PA) is described that allows discrimination between strains harboring flagellin type a or b. All 52 PA strains known to harbor flagellin type b were positive in this ELISA when screened with a fully human monoclonal antibody (LST-007) targeting flagellin type b. Completion of this assay in only 6 h, from picking a single bacterial colony to a colorimetric product, could easily be adapted to a clinical laboratory setting and permit the appropriate choice of therapeutic monoclonal antibody versus its homologous flagellin target in PA-infected patients. [Copyright &y& Elsevier]

Bioassay guided fractionation of an antimycobacterial extract of Foeniculum vulgare var dulce (Apiaceae) led to the isolation and characterization of 5-hydroxyfuranocoumarin. The chemical structure of this compound was elucidated by 1H and 13C (1D and 2D) Nuclear Magnetic Resonance (NMR) spectroscopy. In addition, the active fractions were analyzed by GC-MS and seventy eight compounds were identified; the major compounds were 1,3-benzenediol, 1-methoxycyclohexene, o-cymene, sorbic acid, 2-hydroxy-3-methyl-2-cyclopenten-1-one, estragole, limonene-10-ol and 3-methyl-2- cyclopenten-1-one. Twenty compounds identified in the active fractions were tested against one sensitive and three MDR strains of Mycobacterium tuberculosis using the Alamar Blue microassay. Compounds that showed some degree of antimycobacterial activity against all strains tested were the following: linoleic acid (MIC 100 μg/mL), oleic acid (MIC 100 μg/mL), 1,3-benzenediol (MIC 100-200 μg/mL), undecanal (MIC 50-200 μg/mL), and 2,4-undecadienal (MIC 25-50 μg/mL), the last being the most active compound. To our knowledge, this is the first report of the presence of 5-hydroxyfuranocoumarin in F. vulgare. [ABSTRACT FROM AUTHOR]

Pseudomonas aeruginosa is a Gram-negative bacterium which is capable of developing a high level of antibiotic resistance. It has been placed on the WHO's critical priority pathogen list and it is commonly found in ventilator-associated pneumonia infections, blood stream infections and other largely hospital-acquired illnesses. These infections are difficult to effectively treat due to their increasing antibiotic resistance and as such patients are often treated with antibiotic combination regimens. Methods: We conducted a systematic search with screening criteria using the Ovid search engine and the Embase, Ovid Medline, and APA PsycInfo databases. Results: It was found that in many cases the combination therapies were able to match or outperform the monotherapies and none performed noticeably worse than the monotherapies. However, the clinical studies were mostly small, only a few were prospective randomized clinical trials and statistical significance was lacking. Conclusions: It was concluded that combination therapies have a place in the treatment of these highly resistant bacteria and, in some cases, there is some evidence to suggest that they provide a more effective treatment than monotherapies. [ABSTRACT FROM AUTHOR]

P-glycoprotein (P-gp), the human multidrug resistant (MDR1) gene product and cancer multidrug resistance-associated adenosine triphosphate (ATP)-binding cassette (ABC) transporter, is physiologically expressed on peripheral blood mononuclear cells, but its role in cellular immunity is only beginning to be elucidated. A role of P-gp in the secretion of several T-cell and antigen presenting cell-derived cytokines has been described, and additional functions of the molecule have been identified in lymphocyte survival and antigen presenting cell differentiation. Taken together, these findings provide compelling evidence that P-gp serves several distinct functions in the initiation of primary immune responses, and a critical role of the molecule in functional alloimmune responses is now established. Here, we will review the current understanding of P-gp function in alloimmune T-cell activation via both T-cell and antigen presenting cell-dependent mechanisms, which is relevant to the field of clinical transplantation, where P-gp has been found to be a marker of acute and chronic allograft rejection. Indeed, current in vitro findings raise the possibility that P-gp could represent a novel therapeutic target in acute and chronic allograft rejection, the major causes of allograft dysfunction and ultimate graft loss. [Copyright &y& Elsevier]

Drug resistance, one of the major obstacle in the successful anticancer therapy, can be observed at the outset of therapy (intrinsic resistance) or after exposure to the antitumor agent (acquired resistance). To gain a better insight into the mechanisms of intrinsic resistance we have analyzed two human cell types derived from untreated tumors: MCF-7 breast cancer and A549 non small cell lung cancer (NSCLC). We have examined: the cytotoxic effect induced by doxorubicin (DOX); the time course of drug accumulation by flow cytometry and intracellular drug distribution by confocal microscopy; the expression and distribution of proteins related to anthracycline resistance, such as P-gp (P-glycoprotein), MRP1 (multidrug resistance-associated protein) and LRP (lung resistance-related protein). The cytotoxicity assays showed that A549 cells were less sensitive than MCF-7 cells to the DOX treatment in agreement with the different DOX uptake. Moreover, while in A549 cells DOX was mostly located in well defined intracytoplasmic vesicles, in MCF-7 cells it was mainly revealed inside the nuclei. The analysis of P-gp and MRP expression did not show significant differences between the two cell lines while a high expression of LRP was detected at the nuclear envelope and cytoplasmic levels in A549 cells. These findings suggest that the lower sensitivity to DOX treatment showed by lung carcinoma cells could be ascribed to drug sequestration by LRP inside the cytoplasmic compartments. [ABSTRACT FROM AUTHOR]

Antibiotic resistance is rising at a pace that is difficult to cope with; circumvention of this issue requires fast and efficient alternatives to conventional antibiotics. Algae inhabit a wide span of ecosystems, which contributes to their ability to synthesize diverse classes of highly active biogenic metabolites. Here, for the first time, we reviewed all possible algal metabolites with broad spectra antibacterial activity against pathogenic bacteria, including antibiotic-resistant strains, and categorized different metabolites of both freshwater and marine algae, linking them on the basis of their target sites and mechanistic actions along with their probable nanoconjugates. Algae can be considered a boon for novel drug discovery in the era of antibiotic resistance, as various algal primary and secondary metabolites possess potential antibacterial properties. The diversity of these metabolites from indigenous sources provides a promising gateway enabling researchers and pharmaceutical companies to develop novel nontoxic, cost-effective and highly efficient antibacterial medicines. Unlabelled Image • The summary of the different types of algal metabolites isolated from fresh and marine water algae. • Nano-conjugates modified using these metabolites are reviewed. • Antibacterial activities of these are discussed. [ABSTRACT FROM AUTHOR]