Extended-spectrum Beta-lactamase Research Articles

Highly resistant Salmonella Heidelberg circulating in broiler farms in southern Brazil.

Salmonella Heidelberg, a serotype commonly found in Southern Brazil, is characterized by its high resistance and persistence in the poultry production. This study aimed to characterize the antimicrobial resistance of S. Heidelberg strains. In total, 100 strains isolated from poultry between 2020 and 2022 were evaluated. Phenotypic analyses were performed to determine the susceptibility of 16 antimicrobial agents and detect extended-spectrum beta-lactamase (ESBL)-producing strains. Molecular analyses were performed to detect 11 antimicrobial resistance genes (using polymerase chain reaction [PCR]) and integron class 1 genes (using real-time PCR). A total of 98% of isolates was classified as multidrug-resistant. All isolates were resistant to penicillin and lincomycin. High resistance rates (> 85%) were observed for tetracycline, doxycycline, cephalexin, amoxicillin, and ceftiofur. A significant increase (p < 0.05) in antimicrobial resistance is observed for amoxicillin, cephalexin, and ceftiofur between 2020 and 2022. No significant differences (p > 0.05) were observed in antimicrobial resistance with respect to the region of isolation, season, or company. In total, 25% of isolates were ESBL producers. Integron class 1 gene was detected in only one strain, whereas sul2 was detected in 99%, tet(A) in 66%, blaTEM in 37%, strB in 17%, cmlA in 15%, and tet(B) in 11% of the strains. Other genes were not detected or were detected in < 2% of the strains. The results showed a high overall resistance, which increased over the evaluated period. The high proportions of ESBL-producing and antimicrobial resistant strains represent a risk for highly-resistant S. Heidelberg dissemination across broiler flocks.

Public health concern of antimicrobial resistance and virulence determinants in E. coli isolates from oysters in Egypt

AbstractThe emergence of critical-priority E. coli, carrying a wide array of resistance and virulence factors through food sources, poses a significant challenge to public health. This study aimed to investigate the potential role of oysters sold in Egypt as a source for E. coli, identify their resistance and virulence-associated gene profiles, and assess associated zoonotic risks. A total of 33 pooled fresh oyster samples were obtained from various retail fish markets in Egypt and examined bacteriologically for the presence of E. coli. Antimicrobial resistance was performed by the disk-diffusion method, and the multiple antibiotic resistance index (MAR) was calculated. All isolates were screened for extended-spectrum beta-lactamase (ESBL) (blaTEM, blaSHV, blaCTX−M, and blaOXA−1), plasmid-mediated AmpC blaCMY−2, and carbapenemases (blaKPC, blaNDM, blaVIM, and blaOXA−48) genes by Polymerase chain reaction. Moreover, the presence of virulence-encoding genes was investigated. The virulent MDR strains were clustered using R with the pheatmap package. The prevalence of E. coli was 72.7% (24 out of 33), with 66.7% of the isolates classified as multi-drug resistant, and 75% exhibited MAR values exceeding the 0.2 threshold. Different antimicrobial sensitivity phenotypes and genotype profiles were identified in E. coli isolates. The most prevalent gene detected among all isolates was blaTEM (22/24, 91.7%). Notably, all non-ESBL producers were positive for blaCMY2. Carbapenem-resistant and carbapenem-intermediate strains were carbapenemase producers, with the predominance of the blaKPC gene (11/24, 45.8%). Remarkably, twelve out of sixteen virulence genes were identified, with papC (21/24, 87.5%) and sfa (16/24, 66.7%) genes being the most prevalent. Most isolates carry virulence genes primarily associated with extra-intestinal pathogenic E. coli (ExPEC) (87.5%) and enteropathogenic (EPEC) (70.8%) pathotypes. Four E. coli isolates exhibit cluster patterns. This study provides the first insight into the emergence of virulent MDR E. coli among oysters in Egypt. It underscores the potential role of oysters as a source for disseminating these strains within aquatic ecosystems, presenting a possible threat to public health.

Characterization of Extended Spectrum Beta-lactamase Uropathogens Isolated from Refugees with Urinary Tract Infections in Nakivale Refugee Settlement Camp, Southwestern Uganda

The World Health Organization estimates one in four individuals has had at least one urinary tract infection (UTI) episode requiring treatment with an antimicrobial agent. At Nakivale refugee camp, the overwhelming number of refugees often associated with poor living conditions predispose the refugees to urinary tract infections. This study determined the prevalence of UTIs, the antimicrobial susceptibility pattern of the isolated bacterial pathogens, the prevalence of Extended-Spectrum Beta-Lactamase (ESBL) bacteria, and the molecular characterization of genes encoding ESBLs among refugees in the Nakivale refugee settlement. This was a cross-sectional study that involved 216 outpatients who visited Nakivale Health Centre III between July and September 2020. The urine samples were received and examined at the microbiology laboratory of Mbarara University of Science and Technology. The urine samples were cultured and identified. Antibiotic susceptibility was carried out following CLSI recommended guidelines while the presence of genes encoding ESBL was detected using conventional PCR amplification. The prevalence of UTI was 24.1% (52/216). Staphylococcus aureus was the most prevalent causative agent, accounting for 22/52 (42.31%) of total isolates, followed by Escherichia coli 21/52(40.38%). Multidrug-resistant isolates accounted for 71.15% (37/52). A total of twenty-one isolates (70.0%) were extended spectrum beta-lactamase producing bacteria. The most prevalent genes were TEM beta-lactamase (blaTEM) and CTX-M beta-lactamase (blaCTX-M). The prevalence of UTI among refugees in the Nakivale settlement was high which calls for continuous epidemiological surveys to determine the prevalence of multi-drug resistance uropathogens including ESBL-producing organisms across refugee camps in Uganda.

Analysis of molecular mechanisms of delafloxacin resistance in Escherichia coli

In this study delafloxacin resistance mechanisms in Escherichia coli strains were analyzed. Delafloxacin is a new fluoroquinolone, that is approved for clinical application however, resistance against this agent is scarcely reported. In our study 37 E. coli strains were included and antimicrobial susceptibility testing was performed for ciprofloxacin, delafloxacin, levofloxacin, moxifloxacin, ceftazidime, cefotaxime, imipenem. Six delafloxacin resistant E. coli strains were selected for whole-genome sequencing and all of them exhibited resistance to other fluoroquinonlones and showed an extended-spectrum beta-lactamase phenotype. The six delafloxacin resistant E. coli strains belonged to different sequence types (STs) namely, ST131 (2 strains), ST57 (2 strains), ST162 and ST15840. Each delafloxacin resistant strain possessed multiple mutations in quinolone resistance-determining regions (QRDRs). Notably, three mutations in gyrA Ser83Leu, Asp87Asn and parC Ser80Ile were in strains of ST162, ST57 and ST15840. However, the two strains of ST131 carried five combined mutations namely, gyrA Ser83Leu, Asp87Asn, parC Ser80Ile, Glu84Val, parE Ile549Leu. Association of delafloxacin resistance and production of CTX-M-15 in ST131, CMY-2 in ST162 and ST15840 was detected. In this study a new ST, ST15840 of clonal complex 69 was identified. Our results demonstrate, that at least three mutations in QRDRs are required for delafloxacin resistance in E. coli.

"Molecular Characterization of Extended Spectrum Beta-Lactamase Resistance in Pediatric Shigella Isolates in Egypt".

Shigellosis is a major cause of morbidity and mortality among children, especially in developing countries. The increased extended-spectrum beta-lactamase (ESBL) resistance in Shigella poses a challenge for effective treatment. To examine the antibiotic resistance and ESBL profile of Shigella isolates from children with acute diarrhea. Shigella was isolated from stool cultures from pediatric patients suffering from acute diarrhea. The isolates were identified by bacteriological tests, serotyping, and multiplex polymerase chain reaction (PCR). The antimicrobial resistance was examined by disc diffusion. Phenotypic tests and PCR examined the ESBLs and CTX-M, SHV, and TEM genes. A total of 100 Shigella (10% prevalence rate) were isolated. The S. sonnei and S. dysenteries were the most prevalent species (33% and 31%, respectively), followed by S. flexneri (27%), and only 9% were S. boydii. The isolates had complete resistance (100%) to ampicillin. There was lower resistance to ciprofloxacin (24%), and no resistance to imipenem. By phenotypic tests, 54% of isolates had ESBL. By PCR, bla-CTX-M gene was the most prevalent (50%), followed by bla-TEM (48.1%). Only one isolate (1.9%) had the bla-SHV gene. The alarmingly high rates of antibiotic resistance and ESBL resistance among Shigella spp highlight the urgent need to restrict the unguided use of these drugs. Continuous monitoring of local and global antibiotic resistance patterns is required to prevent the spread of resistance.

Varicella challenges: A case of respiratory tract complications in an elderly patient

Varicella, caused by the varicella-zoster virus (VZV), is rarely reported in the elderly but often complicates with pneumonia. In this case report, we present a case of varicella pneumonia in the elderly. A 60-year-old man presented to the emergency room with vesicles filled with clear fluid that had appeared all over the body for the past four days. The patient also reported fever, headache, pain when swallowing, and itching. Initially, the rash consisted of pink macules that progressed to papules, vesicles, pustules, and crusts. The patient experienced a cough and shortness of breath one day before the hospital visit but did not report any chest or abdominal pain. Notably, the patient’s two grandchildren had a history of chickenpox. The patient had been living with diabetes for five years and was undergoing treatment with insulin injections. The patient was also an active smoker for 40 years. On physical examination, several vesicles with varying degrees of redness were observed, along with yellow-black crusts, pustules, and erosions. Diagnostic tests revealed multinucleated giant cells on a Tzanck smear; pulmonary inflammation with thickening of the right hilum on a chest X-ray; extended-spectrum beta-lactamases Klebsiella pneumoniae in the sputum culture; and elevated hemoglobin A1c, blood sugar level, and liver and kidney function markers on laboratory examination. Based on the clinical findings, the patient was diagnosed with varicella, community-acquired pneumonia, diabetes, acute renal failure, and elevated transaminase enzymes. The patient was treated with acyclovir, paracetamol, salicylate 2% powder, fusidic acid 2% cream, moxifloxacin, N-acetylcysteine, curcuma, and insulin. The patient’s clinical condition improved, the complaint of dyspnea decreased, and the patient was discharged to outpatient care after eight days of hospitalization. This case highlights that while varicella is rare in the elderly, it often presents with severe varicella pneumonia, making early detection and comprehensive management essential to prevent complications and mortality.

Prevalence, Risk Factors and Antibiotic Resistance of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli in Children Hospitalized with Urinary Tract Infection at King Abdulaziz University Hospital, Jeddah, Saudi Arabia

Background/Objectives: We aimed to assess the prevalence and risk factors for acquisition of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (E. coli) in children admitted with urinary tract infection (UTI) at a tertiary university hospital in Saudi Arabia, as well as to investigate antibiotic resistance patterns. Methods: This retrospective cross-sectional study involved hospitalized children aged 0–14 years from January 2018 to December 2022 with urine cultures that grew E. coli or ESBL-producing E. coli. Data of the antimicrobial susceptibility for isolated bacteria were collected. Results: This study analyzed 242 urine samples obtained from 119 children with E. coli UTIs. Of these, 20.7% (n = 50) were ESBL producers. Previous antibiotic use (last 3 months), prophylactic antibiotic use, prior UTI (last 3 months), recurrent UTIs, and underlying co-morbidities (p = 0.011, &lt;0.001, 0.025, &lt;0.001, and 0.013, respectively) had a significant relationship with increased risk of ESBL E. coli UTIs. Generally, the highest resistance rates in the ESBL-producing isolates were for ampicillin and third-generation cephalosporin. Conversely, all ESBL-positive isolates were sensitive to meropenem, with variable resistance rates to other antibiotics as amikacin, nitrofurantoin, quinolones and trimethoprim/sulfamethoxazole (2%, 8%, 56% and 64%, respectively). Conclusions: There is a high prevalence of ESBL production among children hospitalized with E. coli UTIs. Addressing ESBL UTI risk factors helps to recognize high-risk cases and enhance proper antibiotic use.

First-Genome Sequence Data of an Alcaligenes nematophilus Strain EBU-23 Encoding bla Gene Implicated in Acute Childhood Gastroenteritis.

Although an environmental contaminant, Alcaligenes sp. are now increasingly reported in clinical infections. Here, we present a whole-genome sequence of an extended spectrum beta-lactamase (ESBL) Alcaligenes nematophilus strain EBU-23 encoding beta-lactamase, bla gene, isolated from the stool of a hospitalized infant with acute gastroenteritis in a tertiary hospital in Nigeria. Antibiotics susceptibility test revealed that the isolate was resistant to the β-lactams, cephalosporins, and penicillin, β-lactam combination, and the fluoroquinolones. Genomic analysis exposed the presence of a class A beta-lactam hydrolase gene and multiple multidrug efflux permease which may be responsible for the beta-lactamase and multidrug resistance observed with the isolate. To the best our knowledge, we describe the first whole-genome sequence of a multidrug-resistant A. nematophilus implicated in acute childhood gastroenteritis in Nigeria.

New Insights into the Antibacterial Activity of Hydroxytyrosol Extracted from Olive Leaves: Molecular Docking Simulations of its Antibacterial Mechanisms.

This study investigated the biological activities of a hydroxytyrosol-rich extract from Olea europaea leaves, particularly its ability to eradicate severe pathogenic bacteria producing Extended-Spectrum Beta-Lactamases (ESBLs). The latter bacteria are emerging microorganisms that pose significant challenges due to their resistance to a broad range of potent therapeutic drugs. The extract was prepared through an accessible acid hydrolysis method. In vitro and In silico analyses through MIC, MBC analysis and molecular docking were conducted to evaluate the antibacterial properties. The extract showed remarkable antioxidant activity and significant antibacterial potential against reference species and ESBL bacteria. MIC and MBC calculations confirmed the extract's capacity to kill bacteria rather than just inhibit their growth. Further in silico analyzes demonstrated the high binding affinity of HT to the active sites of the gyrase B subunit and the peptidoglycan DD-transpeptidase domain from proteins located in the cytoplasm and the cell wall of the bacteria, respectively. Results confirmed the structure-activity relationship and the ability of HT to disrupt essential bacterial functions. This study validates the debated antimicrobial potential of HT and highlights its importance as a potential therapeutic agent against resistant bacteria, which is a critical area of research given the global challenge of antibiotic resistance.

Genetic profile of carbapenem-resistant &lt;i&gt;Acinetobacter baumannii&lt;/i&gt; strains

The purpose of the research: the molecular genetic characteristic ofA. baumanniicarbapenem-resistant strains, including clonal affiliation, resistome and virulome pattern analysis, description of genetic environment resistance determinants, comparative genetic analysis, and assessment of phylogenetic relationships are presented in the work. The studiedA. baumanniistrains belonged to sequence types ST2Pas/ST2062.2063Oxf(n = 5) and ST78Pas/ST1757Oxf(n = 2). The nucleotide sequences of the studiedA. baumanniiST2Pas/ST2062.2063Oxfstrains were grouped into a single cluster according to phylogenetic analysis. And the sequences of theA. baumanniiST78Pas/ST1757Oxfstrains were combined with the nucleotide sequence of theA. baumanniiAbCTX5 strain, isolated in France in 2015. The presence of intrinsic (OXA-51-like) and acquired carbapenemases genes was shown inA. baumanniistrains. In particular,blaOXA-23are identified in members of ST2Pas/ST2062.2063Oxf, and inA. baumanniiST78Pas/ST1757Oxfstrains —blaOXA-72as part of the plasmid DNA. TheA. baumanniiST78Passtrains possessed additional extended-spectrum beta-lactamase genes. Тhe CTX-M-115 cephalosporinase gene are present in both strains, and theA. baumanniistrain NNAb_2023.3 has theblaCARB-16gene. MostA. baumanniistrains are characterized by the presence of acquired genes for enzymatic modification of macrolides(mph(E), msr(E)), chloramphenicols(catB8), aminoglycosides(aadA, aph(3'')-Ib, aph(6)-Id, aac(6')-Ib, aph(3')-Ia, armA). A comparative analysis showed that inA. baumanniiST2Pas/ST2062.2063Oxfstrains the resistance determinants to macrolides and aminoglycosides are located in the Tn6279-like transposon, and the aminoglycosidases genesaph(3'')-Ib, aph(6)-Idare associated with theIS91-like mobile element. InA. baumanniiST78Pas/ST1757Oxfstrains, resistance genes to aminoglycosides, macrolides, sulfonamides, and beta-lactamase genes are grouped in a region from 60 to 80 Kb long between theglmSandhutCgenes. The presence of mutations in thegyrAandparCgenes associated with resistance to fluoroquinolones were characterized in allA. baumanniistrains. Thus, new knowledge about the genetic profile of carbapenem-resistantA. baumanniistrains representing epidemically significant international clonal lineage has been obtained.

Antibiotic resistance and the search for resistance and virulence genes in Escherichia coli strains isolated from agropastoral farms (Poultry, Cattle, and Fish) In Southern Benin

Resistance of animal-origin enterobacteria to antibiotics is experiencing a concerning global increase, with an escalating impact of extended-spectrum beta-lactamases (ESBL). Finding solutions to this problem requires reliable and diverse data. It is in this context that the present study was initiated to combat infections caused by Escherichia coli in agropastoral farms (poultry, cattle, and fish) in southern Benin. Samples consisting of cattle dung, poultry droppings, as well as water and sludge from fish farms were aseptically collected for various laboratory analyses. A total of 714 E. coli strains were isolated from 1,376 collected samples, with 434 strains from the Atlantique department and 280 strains from the Zou department. The isolation of strains was carried out according to the NF V08-050: 2009 standard. The identified strains were tested against various antibiotics using the Bauer and Kirby disk diffusion method on agar, as recommended by the WHO. Strains resistant to one or more antibiotics were selected for the search for resistance and virulence genes. Furthermore, multidrug resistance of the strains to antibiotics was also observed, particularly in the Beta-lactam class (100% resistance to AMC) and tetracyclines (90% resistance to tetracyclines). Additionally, genotypic detection revealed a total absence of virulence genes, whereas resistance genes such as beta-lactamase Sulfhydryl Variable (bla-SHV), beta-lactamase Temoniera (bla-TEM), and quinolone resistance gene B (qnrB) were more frequently detected at respectively 58%, 82% and 60%.

Current antibiotic resistance of Enterobacteriaceae isolated at Avicenna military hospital of Marrakesh

Enterobacteriaceae are bacteria found in the digestive system, known for causing infections and displaying antibiotic resistance. The World Health Organization (WHO) identifies strains resistant to third-generation cephalosporins and carbapenems as priority pathogens. This study, conducted at Avicenne Military Hospital of Marrakech, aimed to assess the epidemiological and antibiotic resistance profiles of Enterobacteriaceae over a two-year period (May 2020 - April 2022) to inform updated antibiotic use recommendations. This retrospective study analyzed 1,626 positive bacterial isolates from various hospital departments, including medical, surgical, intensive care, emergency units, and external samples. Of these, 1,163 (71.5%) were Enterobacteriaceae, predominantly Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae, primarily isolated from urine, pus, and blood cultures. Antibiotic resistance was assessed, revealing that 614 (52.8%) of Enterobacteriaceae were extended-spectrum beta-lactamase producers (ESBL-E), and 115 (9.9%) were carbapenemase-producing Enterobacteriaceae (CPE). The study’s findings align with existing literature, showing a higher prevalence of Enterobacteriaceae in males and frequent isolation of Escherichia coli. The rise in multidrug-resistant Enterobacteriaceae, particularly ESBL-E and CPE, poses risks such as therapeutic failure, increased morbidity and mortality, and significant economic impacts. To address this, the study emphasizes the importance of early detection, infection control measures, responsible antibiotic prescribing, and collaborative efforts across hospital departments to reduce the spread of resistant strains.

Spatial and temporal dynamics of the prevalence of resistance genes and gastrointestinal pathogens in stool samples of German deployment returnees

AbstractBackgroundThe exploratory study assessed trends in the abundance of CTX-M-type extended spectrum beta-lactamase (ESBL) and vancomycin-resistance genes vanA and vanB in the stool samples of German soldiers and police officers returning from predominantly tropical deployments next to the common diarrheagenic Escherichia (E.) coli pathovars enteropathogenic Escherichia coli (EPEC), enterotoxigenic E. coli (ETEC) and enteroaggregative E. coli (EAEC)) as well as rarely imported Vibrio spp. between 2006 and 2024.MethodsSurveillance was performed applying real-time polymerase chain reaction and results were stratified by World Health Organization region of deployment as well as by deployment period. For the latter, the study interval was divided into three pre-COVID-19-pandemic periods, the COVID-19-pandemic period and the post-COVID-19-pandemic period. Averaged prevalences were used as references.ResultsIn stool samples of 1817 deployed German soldiers and 117 police officers, averaged prevalences were 47.9% and 24.8% for the ESBL-type beta-lactamase blaCTX-M, 30.2% and 14.5% for vanB, 9.0% and 17.9% for EPEC, 3.4% and 12.8% for ETEC, 4.0% and 3.4% for EAEC as well as 2.0% and 3.4% for Vibrio spp., respectively. While resistance genes peaked during early deployments, maximum prevalences for enteropathogens were seen later.ConclusionsThe assessment suggested time- and region-dependence of the assessed parameters.

A single-center analysis of clonal transmission of carbapenem-resistant Acinetobacter baumannii among intensive care unit patients during the COVID-19 pandemic

Carbapenem-Resistant Acinetobacter baumannii (CRAB) outbreak in intensive care units (ICUs) is a significant problem for healthcare facilities. In this study, we aimed to investigate the occurrence of CRAB isolates among ICU-admitted patients during the three waves of the COVID-19 pandemic in Iran using Multiple-Locus Variable Number Tandem-Repeat Analysis (MLVA). We obtained 50 (A) baumannii isolates from tracheal aspirate and blood culture samples. In the disc diffusion method, all isolates were cefotaxime, ceftriaxone, and cefepime-resistant, while 98% (49/50) of isolates were resistant to piperacillin, piperacillin-tazobactam, ceftazidime, and ciprofloxacin. Levofloxacin and tobramycin resistance was found in 76% (38/50) of isolates. In the microbroth dilution test all isolates were resistant to imipenem, 98% (49/50) to meropenem, 68% (34/50) to colistin, and 20% (10/50) to polymyxin (B) Based on the PCR findings, all isolates harbored blaOXA−40, ISAba-1, and int-2 genes. There were no isolates found that have the blaOXA−58, blaOXA−143, blaVEB−1, blaVIM, and int-3 genes. Among Extended-spectrum beta-lactamases (ESBL) genes, blaCTX−M, blaTEM, blaSHV, blaGES, and blaPER−1 have a prevalence of 42% (21/50), 84% (42/50), 58% (29/50), 78% (39/50), and 54% (27/50), respectively. 74% (37/50) of the isolates had the blaOXA−23 gene, while all of the isolates carried the blaOXA−40 gene. Among MBL genes, blaIPM, blaGIM, blaSIM, and blaNDM−1 have a prevalence of 20% (10/50), 8% (4/50), 22% (11/50), and 60% (30/50), respectively. The prevalence of int-1 was documented as 74% (37/50). Accordingly, all isolates were identified as CRAB. The co-existence of blaOXA−23/int-2 and blaOXA−23/isaba-1 was 74% (37/50). The co-existence of blaNDM−1/ISAba-1 was observed in 30 (60%) isolates. Using an 80% similarity threshold on the dendrogram constructed through MLVA typing, all isolates were grouped into two clusters: cluster A with 9 isolates from wave 3, and cluster B with 41 isolates from waves 3, 4, and 5. Our study confirms a clonal transmission of CRAB during the study period and suggests using molecular typing methods like MLVA in healthcare settings to identify dominant clones, antibiotic resistance patterns, and transmission routes. This will help to better manage the emergence and spread of antibiotic-resistant strains in future outbreaks.

Occurrence, Multidrug Resistance, and Multilocus Sequence Typing of Extended-Spectrum Beta-Lactamase/AmpC-Producing Escherichia coli from Farmed Eggs.

The present study evaluated the occurrence, antibiogram profile, and sequence types (STs) of multidrug resistant (MDR) Escherichia coli from freshly laid eggs (n = 480), feed (n = 24), water (n = 24), poultry droppings (n = 24), and hand swab samples (n = 10) collected from 24 deep litter (DL) and caged poultry layer farms (12 per category) across Punjab, India. The overall E. coli contamination rate in DL and cage farms was 32% (95% confidence intervals [CI], 26.6-37.8%) and 16.7% (95% CI, 12.6-21.6%), respectively. The logistic regression analysis revealed that the DL system had higher odds of occurrence (odds ratio [OR]) of extended-spectrum beta-lactamase (ESBL) (2.195, 95% CI, 1.065, 4.522) and ESBL/AmpC coproducers (2.69, 95% CI, 1.122, 6.45) compared to the cage system. Additionally, isolates from the DL were 4.065 (95% CI, 1.477, 11.188) times more tetracycline resistant compared to the latter; however, resistance to amoxyclavulanate (OR, 0.437; 95% CI, 0.209, 0.912), and ampicillin (OR, 0.343; 95% CI, 0.163, 0.720) was lesser in DL system. Notably, around 97.7% and 87.2% of the isolates from the DL and cage system were MDR, with the DL system having 6.439 (95% CI, 1.246, 33.283) times more chances of harboring MDR E. coli. Additionally, among the resistance genes, the DL system demonstrated significantly high presence of blaAmpC (56%), qnrA/B/S (42.3%), and tetA/B (30.6%). Furthermore, multilocus sequence typing of 11 MDR isolates (n = 5, DL, and 6, cage) revealed the presence of 10 STs, of which ST10, ST155, and ST156 were found to be of public health importance. Therefore, the present study highlights the burden of MDR, ESBL, and AmpC-producing E. coli on poultry eggs and farm environment, which could be carried over to human handlers and consumers upon direct contact during handling and processing.

Water Quality and Prevalence of Extended Spectrum Beta Lactamase Producing Escherichia coli (ESBL E. coli) in Sungai Terengganu, Malaysia

Antimicrobial resistance (AMR) is a significant global issue impacting human, animal, and environmental health, as well as economic security. Water bodies, especially, are now recognized for their role in the spread of antimicrobial-resistant pathogens, though standardized monitoring methods are lacking. This study investigates the presence of extended-spectrum beta-lactamase (ESBL) producing Escherichia coli in the Sungai Terengganu River in Malaysia, focusing on water quality and ESBL E. coli monitoring. Sampling was conducted twice in 2023 at 13 locations along the river. Overall, the water quality index (WQI) classified the river as Class II (WQI = 81), with upstream regions rated as Class I, indicating minimal anthropogenic impact. Higher Biological Oxygen Demand (BOD), Chemical Oxygen Demand (COD), and Total Suspended Solids (TSS) in downstream areas suggested organic pollution from anthropogenic activities. Four metal elements exceeded the National Water Quality Standards (NWQS): Al (0.88 mg/L), Fe (3.45 mg/L), Cu (0.102 mg/L), and Hg (2.525 mg/L). No Pb, Cr, Se, Sn, or As were detected. Statistical analysis showed better water quality correlates with lower microbial contamination. ESBL E. coli was found in 8 of 13 sampling sites (61.5%), with the highest concentration at 800 CFU/100 mL. From 40 presumptive ESBL isolates tested, 87.5% (35 isolates) were positive for ESBL and carried additional drug resistance genes, including mcr-1, tetW, sul-1, and sul-2. These findings indicate a concerning presence of multidrug-resistant E. coli in the river, representing a potential public health risk.

Risk factors associated with multidrug-resistant Klebsiella pneumoniae infections: a multicenter observational study in Lebanese hospitals

BackgroundKlebsiella pneumoniae is a significant global public health burden, especially in low-income countries and regions with fragile healthcare infrastructures, due to its ability to cause severe infections, increase mortality rates, and its rising antimicrobial resistance. This study aimed to estimate the proportion of multidrug-resistant (MDR) K. pneumoniae infections and identify associated risk factors.MethodsData were retrospectively collected from three academic hospitals in Beirut, Lebanon, between January 2021 and September 2023 using a standardized form. Binary logistic regression was used to determine risk factors associated with MDR, extended-spectrum beta-lactamase (ESBL)-producing, and carbapenem-resistant K. pneumoniae (CRKP) infections.ResultsOut of 2,655 K. pneumoniae cases, 410 met the inclusion criteria. The primary infection sources were the urinary tract (58.3%) and the respiratory tract (12.4%). Among the isolates, 61% were MDR K. pneumoniae, with 7.3% being extensively drug-resistant, and 0.5% pandrug-resistant. Additionally, 36.8% were ESBL-producing, while 6.3% were CRKP. Predictors significantly associated with MDR K. pneumoniae infections included male sex (adjusted odds ratio [AOR] = 3.46, 95% CI = 1.01–11.86, P = 0.04), recent antibiotics use (AOR = 4.52, 95% CI = 1.65–12.36, P = 0.003), and recent cancer chemotherapy (AOR = 3.43, 95% CI = 1.25–9.42, P = 0.01). ESBL-producing infections were associated with age ≥ 65 years, higher Charlson Comorbidity Index (CCI), and recent antibiotic use. CRKP infections were linked to male sex, prior antibiotic use, and longer hospital stays prior to infection (all P < 0.05).ConclusionsMDR K. pneumoniae infections are steadily rising in Lebanon, along with an increase in ESBL-producing and CRKP cases. The main risk factors for MDR K. pneumoniae infections were male sex, recent antibiotic use, and cancer chemotherapy. ESBL-producing infections were associated with advanced age, higher CCI, and recent antibiotic use, while CRKP infections were linked to male sex, prior antibiotic use, and prolonged hospital stays. This situation is further exacerbated by inadequate healthcare infrastructure and suboptimal national surveillance. Strengthening local surveillance and implementing effective antibiotic stewardship programs are critical to managing this growing threat..

Extended-spectrum beta-lactamase- and carbapenemase-producing Escherichia coli isolates causing hospital- and community-acquired infections in Tunisia (2001-2019): expansion of CTX-M-15-C2 and CTX-M-27-C1 ST131 subclades.

We aimed to investigate the microbiological features of extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli (ESBL-EC) and carbapenemase-producing E. coli (CP-EC) causing hospital- and community-acquired infections in Tunisia over the last two decades. The study captured the emergence and expansion of the CTX-M-15-C2 ST131 subclade and successively the CTX-M-27-C1 ST131 subclade, which were responsible for the steady increase in the prevalence of ESBL-EC. However, the incidence of CP-EC remained stable over the study period with a highly diverse content in carbapenemase genes dominated by blaOXA-48-like. This is the first study to provide comprehensive data on the epidemiology of ESBL-EC and CP-EC in a North African country.

Living without pain: A 10-year study of congenital insensitivity to pain with anhidrosis.

Congenital Insensitivity to Pain with Anhidrosis (CIPA) is a rare hereditary neuropathy caused by NTRK1 gene mutations, predisposing patients to recurrent infections and chronic wounds. Long-term studies on microbial and clinical outcomes in CIPA are limited. This study presents analysis of infection patterns, antibiotic resistance, and clinical outcomes in CIPA patients. A comprehensive ten-year retrospective cohort study was conducted at Soroka University Medical Center, Beer Sheva, Israel, from January 2014 to January 2023. Electronic medical records were reviewed to identify 63 CIPA patients, all were of consanguineous Bedouin families. Data collection included demographic details, clinical presentations, genetic analysis, documentation of infections, wound sites, hospital duration, and surgical interventions. Staphylococcus aureus, notably methicillin-resistant, dominated, with Gram-negative bacteria common in lower limbs. The study noted reduced extended-spectrum beta-lactamase bacteria and linked demographic factors to infection traits, antibiotic resistance, and surgical needs. This study provides valuable insights into the clinical and microbial patterns of CIPA, highlighting dynamic shifts in microbial compositions and antibiotic resistance profiles over time. Staphylococcus aureus, and Gram-negative bacteria particularly in lower limb infections, pose significant challenges in patient management. The findings underscore the importance of tailored approaches to address evolving microbial profiles and optimize patient care in CIPA. Key Message: This is the largest cohort study on CIPA to date, highlighting the dominance of Staphylococcus aureus, including methicillin-resistant strains, and significant Gram-negative bacterial infections in lower limbs. Contribution to Literature: It draws parallels between infection dynamics in CIPA and diabetic foot ulcers, emphasizing similar challenges due to neuropathy and ischemia, enhancing understanding of infection susceptibility and management in neuropathic conditions. The findings inform clinical practices by detailing infection and resistance patterns, supporting the development of targeted treatment strategies to improve outcomes for CIPA and similar conditions.

A novel machine-learning aided platform for rapid detection of urine ESBLs and carbapenemases: URECA-LAMP.

Pathogenic gram-negative bacteria frequently carry genes encoding extended-spectrum beta-lactamases (ESBL) and/or carbapenemases. Of great concern are carbapenem resistant Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Despite the need for rapid AMR diagnostics globally, current molecular detection methods often require expensive equipment and trained personnel. Here, we present a novel machine-learning-aided platform for the rapid detection of ESBLs and carbapenemases using Loop-mediated isothermal Amplification (LAMP). The platform consists of (i) an affordable device for sample lysis, LAMP amplification, and visual fluorometric detection; (ii) a LAMP screening panel to detect the most common ESBL and carbapenemase genes; and (iii) a smartphone application for automated interpretation of results. Validation studies on clinical isolates and urine samples demonstrated percent positive and negative agreements above 95% for all targets. Accuracy, precision, and recall values of the machine learning model deployed in the smartphone application were all above 92%. Providing a simplified workflow, minimal operation training, and results in less than an hour, this study demonstrated the platform's feasibility for near-patient testing in resource-limited settings.IMPORTANCEExtended-spectrum beta-lactamases (ESBL) and carbapenemases confer resistance to third-generation cephalosporins and carbapenems in pathogenic Gram-negative bacteria such as Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, and Acinetobacter baumannii. Conventional antimicrobial susceptibility testing is based on phenotypic methods, and results can take several days to be obtained. Current genotypic detection methods can be rapid but require expensive equipment and trained personnel. In this study, we present a novel machine learning-aided platform for the rapid detection of ESBLs and carbapenemases using Loop-mediated isothermal Amplification (LAMP). The validation of the platform demonstrated percent positive and negative agreements above 95% for all targets. The newly developed platform provided a simplified workflow, minimal technical training, and results in less than an hour. This study demonstrated the platform's feasibility for rapid testing of ESBL and carbapenemases in bacteria and urine specimens.