Extended-spectrum Beta-lactamase Research Articles

Antibacterial Effects of Basil (Ocimum sanctum) Leaf Extract in Combination with Meropenem against Extended Spectrum Beta-Lactamase (ESBL) Producing Klebsiella pneumoniae

Highlights: O. sanctum leaf extract does not significantly increase its antibacterial effects when combined with meropenem against ESBL-producing K. pneumonia. O. sanctum leaf extract has different antibacterial effects depending on geological and environmental factors. Abstract Introduction: Antibiotic usage for bacterial infections causes antibiotic resistance in bacteria. One is extended-spectrum beta-lactamase (ESBL), which produces Klebsiella pneumoniae, a pathogen responsible for increased antibiotic resistance. Basil (Ocimum sanctum) is a candidate for combination therapy. It has been proven to have antibacterial effects. However, its combination with antibiotics is rarely researched. This study evaluated the antibacterial effects of O. sanctum leaf extract and meropenem combination against ESBL-producing K. pneumoniae. Methods: This study used the disk diffusion method. The extract was tested for each experiment at 6%, 4%, 2%, 1%, and 0.5% concentration. The research was divided into two experiments to evaluate the antibacterial effects of O. sanctum leaf extract (n = 18) and its combination with meropenem against ESBL-producing K. pneumoniae (n = 18). The data was analyzed using the International Business Machines Corporation (IBM) Statistical Package for the Social Sciences (SPSS) version 26.0 for Windows. A probability (p) value of < 0.050 was considered significant. Results: The first experiment showed that the extract had the largest antibacterial effect at 0.5% concentration (n = 18, p = 0.007). Meanwhile, the second experiment showed that the combination of the extract and meropenem did not have significant antibacterial effects (n = 18, p = 0.597). Conclusion: O. sanctum leaf extract has viable antibacterial effects, but its combination with meropenem does not significantly improve its antibacterial effects against ESBL-producing K. pneumonia.

Antibacterial Resistance and Extended-Spectrum Beta-Lactamase (ESBL) Phenotypes in Enterobacteriaceae Isolated from Fecal Samples of Humans and Animals in Selected Local Government Areas of Nasarawa State, Nigeria

It is quite alarming the increasing rate of antibacterial resistance all over the world considering the public health threat and the re-emergence of multi-drug resistant Enterobacteriaceae. The aim of this study is Antibacterial resistance and phenotypic detection of Extended Spectrum Beta-Lactamase (ESBL) producing Enterobacteriaceae isolated from human and animal fecal samples in selected local government areas of Nasarawa state, Nigeria was carried out in the study. Hundred (100) samples comprising human and animal (goats, cattle, and chicken) were collected and 55 samples were multidrug resistant. A commercial biochemical kit (Eneterosystem 18R) was used for the isolation and identification of Enterobacteriaceae. Kirby Bauer Disk Diffusion Method was used for antibacterial susceptibility testing of Enterobacteriaceae isolates. The Double Disc Synergy Test (DDST) method was also used for the phenotypic confirmation test of Extended Spectrum Beta Lactamase (ESBL). The occurrence of multidrug-resistant isolates shows that Escherichia coli (100.00%) which is the highest, Proteus mirabilis (14.54%), Klebsiella pneumoniae, and Salmonella enterica (10.90%), while the occurrence of Shigella flexneri (9.09%) was the lowest. The Enterobacteriaceae isolates were more resistant to Cefuroxime, Cefexime, Amoxicillin Clavulanate, and Imipenem/Cilastatin with percentage resistance ranges from 66.6% - 100%. The occurrence of ESBL producers shows that Escherichia coli (60.00%) and Proteus mirabilis (62.5%) were high while Shigella flexneri (20.0%) had a low occurrence of ESBL. The sale and in-discriminate use of antibiotics without a prescription is an important regulatory issue in the abuse of antibiotics for both humans and animals. The Beta-Lactam and gentamycin antibiotics were not effective against the Multi-Drug Resistant (MDR) isolates and most of the isolates were ESBL producers.

Extended-spectrum beta-lactamase (ESBL) production and virulence genes profile of avian pathogenic Escherichia coli (APEC) isolated from broiler chickens in eastern Thailand

Avian pathogenic Escherichia coli (APEC) causes colibacillosis, resulting in extensive economic losses for the broiler industry. To date, there is little information in Thailand about virulence-associated genes and antibiotic resistance of APEC strains. Here, this study aimed to investigate the virulence genes and extended-spectrum beta-lactamase (ESBL) characteristics of APEC isolated from broilers. This study used multiplex polymerase chain reaction (PCR) to determine the presence of virulence genes and resistance genes in APEC. Furthermore, the disc diffusion method examined ESBL phenotypes of APEC and antibiotic resistance profiles against 17 antimicrobials. In this study, E. coli was isolated 11 (6.32%) of 174 broiler visceral organs. All E. coli isolates were tested for five APEC-virulence-associated genes (iroN, ompT, hlyF, iutA, and iss). Eight E. coli isolates from broilers with colibacillosis-associated lesions carried two (iroN and ompT) of the APEC virulence genes. One APEC virulence gene (hlyF) was found in E. coli isolates from broilers without lesions. Possibly no individual virulence gene was specific to the APEC strain. Interestingly, the papC, previously detected in humans with uropathogenic E. coli, was found in an APEC isolate. All APEC isolates were ESBL-producing E. coli, and then they were tested for four beta-lactamase-encoding genes (blaTEM, blaCTX-M, blaOXA, and blaSHV). The blaTEM and blaCTX-M were identified in 81.81% (9/11) of the isolates, whereas blaOXA or blaSHV were not detected in any isolate. All APEC showed multi-drug resistance (MDR) phenotypes, especially chloramphenicol, erythromycin, and sulfamethoxazole-trimethoprim. Although antibiotics were not recently used, MDR might be encouraged by horizontally transferring antibiotic-resistance genes. In addition, fluoroquinolone resistances were found in APEC isolates which could transfer resistance genes to humans via the food chain. This study indicates that APEC isolates contain several virulence and bla genes and should be surveillance to prevent the transmission of those genes to humans.

Comparative analysis of Salmonella contamination in pork slaughtering facilities: Implications for food safety

Salmonella is a major cause of global gastroenteritis, with pig-based products being significant sources. The Hazard Analysis Critical Control Points (HACCP) standard ensures food safety by identifying Salmonella contamination dynamics and improving control strategies. A study in Thailand found that pork in HACCP-certified slaughterhouses had significantly reduced Salmonella contamination and antimicrobial resistance (AMR) compared to in non-HACCP facilities. However, both types of slaughterhouses still experience Salmonella contamination during the slaughtering process, with limited information available on contamination sources and critical steps. The current study compared Salmonella distribution and AMR profiles in two Thai pig slaughterhouses, one of which implemented HACCP. A total of 570 samples, including feces, carcass swabs, and environmental samples, were cultured for Salmonella and investigated for AMR. The overall prevalence was 24.7%, being significantly lower at the HACCP-compliant slaughterhouse 1 (S1, 13.7%) compared to the non-compliant slaughterhouse 2 (S2, 41.2%) (P = 0.03). Slaughterhouse-specific Salmonella prevalence varied across slaughtering steps, with S2 showing high contamination rates in carcasses post-final washing, persisting until post-cutting. Additionally, specific strains were found in various locations, including feces, carcasses, workers' hands, cutting boards, and knives before and during operations. Conversely, carcasses post-final washing in S1 had the lowest contamination. Real-Time PCR identified higher Salmonella contamination and a greater copy number in S2-cutting carcasses compared to in S1. Overall, 42.6% of isolates were multidrug-resistant, showing strong resistance to ampicillin and tetracycline, while 8.51% produced extended-spectrum beta-lactamase (ESBL). It is evident that continuous monitoring and control measures are crucial to mitigate contamination and antibiotic resistance in foodborne pathogens such as Salmonella. The current study confirms that adherence to HACCP in slaughterhouses significantly reduces Salmonella in pork products. However, contamination levels rose notably post-cutting in both HACCP and non-HACCP facilities, with equipment, other carcasses, and workers' hands acting as sources of contamination. Rigorous monitoring during cutting processes is essential to identify and mitigate contamination sources, thereby improving food safety in meat processing facilities.

Diversity and Antibiotic Susceptibility of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae from the Urine of Human Population Using Bacteria-Contaminated Water in Butembo Urban Area (DR Congo, Central Africa)

Aims: This study aims to investigate the extended-spectrum beta-lactamases (ESBLs)-producing Enterobacteriaceae in the human urine samples and to understand the potential influence of the contaminated water the urine donors use. Study Design and Methodology: One hundred urine samples from females and the same number from males were collected from donors of different age groups and analysed. One hundred urine samples from females and the same number from males were collected from donors of different age groups. The enterobacterial isolation was done on CLED agar medium. Their identification was carried out first by the enzymatic technic, then using MALDI–TOF MS method. ESBLs-producing enterobacteria were identified by searching the synergy between Clavulanic acid and 3rd generation Cephalosporins. The antimicrobial susceptibility tests were carried out using the disk diffusion method. Results: In male urine samples, the identified ESBLs-producing strains belong to E. coli, E. agglomerans and K. pneumoniae. They were in 11 samples: 3 from donors aged 20 years or less, 3 also from those aged 21 to 40 years as well as in those aged 41 to 60 years, and 2 in samples from donors aged over than 60 years. In female urine samples, the identified ESBLs-producing strains belong to Escherichia coli, Enterobacter agglomerans, Yersinia frederiksenii, Salmonella typhi and Klebsiella pneumoniae. They were in 27 samples: 3 from donors aged 20 years or less, 9 from those aged 21 to 40 years, 12 from those aged 41 to 60 years, and 3 from donors aged over than 60 years. All the strains were sensitive to Imipenem and resistant to Cefotaxim, Ceftazidin, Amoxicillin+Clavulanic acid and Nitrofurantoin. The Multi-Antibiotic Resistance (MAR) index varied from 0.416 (E. coli strains, youngest male donors) to 0.916 (E. coli strains, old female donors). Most of strains react differently from one antibiotic to another (P<0.05), with the exception of E. coli strains from most samples. Most of the identified ESBLs-producing species have been reported in surface and groundwater the population use. These water resources could play a role in the human urinary tract infections. Conclusion: There is a serious health problem with great MAR indices for all bacterial species identified. Treating water before use could reduce the viability and spreading of ESBLs-producing bacteria and genes into the population.

Salmonella Prevalence and antibiotic resistance profile in raw poultry meat sold in North Lebanon: Insights from the COVID-19 pandemic and economic crisis

Salmonella-related foodborne illness is a significant public health concern, with the primary source of human infection being animal-based food products, particularly chicken meat. Lebanon is currently experiencing a dual crisis: the COVID-19 pandemic and an unprecedented economic crisis, which has resulted in substantial challenges to the public health system and food safety. This study aims to assess the prevalence and antibiotic resistance profile of Salmonella in raw poultry meat sold in North Lebanon during this dual crisis. A cross-sectional study was carried out between May 2021 and April 2022 across six different districts in North Lebanon. A total of 288 whole, unprocessed chickens were examined. The isolation and identification of Salmonella isolates were done based on cultural and biochemical properties. All isolates were subjected to antimicrobial susceptibility testing and phenotypic assays for Extended-Spectrum Beta-lactamase (ESBL) detection. The prevalence of Salmonella in raw poultry meat purchased in North Lebanon reached 18.05 % (52/288). The dry season and chilled chicken were significantly associated with an increased risk of Salmonella contamination (P < 0.05). Additionally, 34.61 % of the isolates were potential ESBL producers, and 57.69 % exhibited multidrug resistance (MDR). This study highlights the existence of MDR in chicken meat in North Lebanon, posing a potential health risk if undercooked chicken meat is consumed. This emphasizes the importance of the implementation of preventive strategies and hygienic procedures throughout the food chain to reduce the risk of Salmonella spp. contamination in chicken meats and its potential transmission to humans.

Molecular Characteristics of Cephalosporin Resistant Escherichia coli and Klebsiella pneumoniae Isolated from Children in a Tertiary Care Centre of Central Kerala, India

Aims: The study was aimed to determine the molecular characteristics of extended-spectrum beta-lactamases (ESBL) producing cephalosporin-resistant Escherichia coli and Klebsiella pneumoniae isolated from children below ten years of age. Background: Geographically diverse variations in the prevalence of ESBL genes were reported. No data were available on the prevalence of ESBL genes in central Kerala, India, among children below 10 years of age. Methods: A cross-sectional study was performed to analyze ESBL genes in cephalosporin-re-sistant E. coli and K. pneumoniae strains isolated from samples received in the Microbiology la-boratory of a tertiary care centre during the period between May 2021 and July 2022. The strains showed that ESBL + cephalosporin resistance was subjected to PCR-based genotyping for the genes such as bla (beta-lactamase) CTX-M-1, blaCTX-M-15, blaCTX-M-U, blaTEM, blaPER and SHV. Results: Among the total 228 samples analyzed, 136 (60%) had no growth. Ninety-two (40 %) samples showed growth of E. coli and K. pneumoniae. Among the isolates that showed growth, 39 (42%) were sensitive, and the remaining 53 (57%) were resistant to third-generation cephalospor-ins. Among the isolates showed resistance, 22 (42%) were ESBL positive and 31 (58%) were ESBL negative. Among the positive ESBL, nine E. coli strains (60%) were positive for CTX-M-15 and CTX-M-1. CTX-M-15 and CTX-M-U were present in six (85%) K. pneumoniae with ESBL +. Conclusion: E. coli and K. pneumoniae isolated from specimens of children below ten years of age showed 41-42% ESBL producers. Prevalent ESBL-producing genes in E. coli were CTX-M-15 and CTX-M-1. CTX-M-15 and CTX-M-U were prevalent in ESBL-producing K. pneumoniae.

Comparing Graph Sample and Aggregation (SAGE) and Graph Attention Networks in the Prediction of Drug-Gene Associations of Extended-Spectrum Beta-Lactamases in Periodontal Infections and Resistance.

Gram-negative bacteria exhibit more antibiotic resistance than gram-positive bacteria due to their cell wall structure and composition differences. Porins, or protein channels in these bacteria, can allow small, hydrophilic antibiotics to diffuse, affecting their susceptibility. Mutations in porin protein genes can also impair antibiotic entry. Predicting drug-gene associations of extended-spectrum beta-lactamases (ESBLs) is crucial as they confer resistance to beta-lactam antibiotics, challenging the treatment of infections. This aids clinicians in selecting suitable treatments, optimizing drug usage, enhancing patient outcomes, and controlling antibiotic resistance in healthcare settings. Graph-based neural networks can predict drug-gene associations in periodontal infections and resistance. The aim of the study was to predict drug-gene associations of ESBLs in periodontal infections and resistance. The study focuses on analyzing drug-gene associations using probes and drugs. The data was converted into graph language, assigning nodes and edges for drugs and genes. Graph neural networks (GNNs) and similar algorithms were implemented using Google Colab and Python. Cytoscape and CytoHubba are open-source software platforms used for network analysis and visualization. GNNswere used for tasks like node classification, link prediction, and graph-level prediction. Three graph-based models were used: graph convolutional network (GCN), Graph SAGE, and graph attention network (GAT). Each model was trained for 200 epochs using the Adam optimizer with a learning rate of 0.01 and a weight decay of 5e-4. The drug-gene association network has 57 nodes, 79 edges, and a 2.730 characteristic path length. Its structure, organization, and connectivity are analyzed using the GCN and Graph SAGE, which show high accuracy, precision, recall, and an F1-score of 0.94. GAT's performance metrics are lower, with an accuracy of 0.68, precision of 0.47, recall of 0.68, and F1-score of 0.56, suggesting that it may not be as effective in capturing drug-gene relationships. Compared to ESBLs, both GCN and Graph SAGE demonstrate excellent performance with accuracy, precision, recall, and an F1-score of 0.94. These results indicate that GCN and Graph SAGE are highly effective in predicting drug-gene associations related to ESBLs. GCN and Graph SAGE outperform GAT in predicting drug-gene associations for ESBLs. Improvements include data augmentation, regularization, and cross-validation. Ethical considerations, fairness, and open-source implementations are crucial for future research in precision periodontal treatment.

Genomic diversity and nutritional analysis of multi-drug resistant extended spectrum β-lactamase Producing-Klebsiella pneumoniae genes isolated from mastitic cattle milk in district peshawar, Pakistan

The increasing incidence of resistance extended spectrum-beta lactamase (ESBL) producing Klebsiella pneumonia become worldwide issue. The current study aimed to determine the genomic diversity of ESBL-producing K. pneumoniae in milk samples collected from cows with mastitis as well as their antibiotic sensitivity profiles and genetic identification in Peshawar, Pakistan. The california mastitis test (CMT) was initially used to verify the presence for mastitis in 700 collected milk samples. The molecular identification of the 16SrRNA gene confirmed 120/700 (17.14 %) propagation of K. pneumonia. Out of these isolates MDR ESBL-producing isolates were 60/120 (50 %). The lactose were found (M = 3.96 ± 0.28, SD = 2.19), followed by fats (M = 3.12 ± 0.11, SD = 0.90), protein (M = 5.97 ± 0.24, SD = 1.84), sodium (M = 55.74 ± 2.07, SD = 15.81), potassium (M = 138.5 ± 1.53, SD = 11.71), chloride (M = 0.74 ± 0.03, SD = 0.24), calcium (M = 10.27 ± 0.31, SD = 2.42), and chlorine (M = 2.80 ± 0.22, SD = 1.70), respectively. Amikacin (80 %), ceftazidime (71 %), and tetracycline (71 %) were shown to be the most effective antimicrobials against all of the isolates. The occurrence of the blaSHV gene was observed at 56.00 % whereas the blaTEM gene and blaCTX-M gene were 36.00 %, and 30.00 %. The distribution of blaCTX-M subgroup genes was followed by blaCTX-M-1 (38.00 %), blaCTX-M-9 (22.20 %), and blaCTX-M-15 (61.10 %). Co-occurrence of blaCTX-M+ blaSHV was (15.00 %), blaCTX-M+ blaTEM were (6.60 %), and blaSHV + blaTEM were (10.00 %), respectively. The inappropriate, prolonged and common use of antibiotics may apply selective pressure for propagation and the occurrence of resistant isolates.

Antibiotic Resistance Profiles of Escherichia coli and Salmonella spp. Isolated From Dairy Farms and Surroundings in a Rural Area of Western Anatolia, Turkey.

Background Antibiotic resistance is a significant public health issue worldwide. Antibiotic-resistant zoonotic bacteria such as Escherichia coli (E. coli), Campylobacter, Salmonella, Listeria, Coxiella, and Mycobacterium can be particularly isolated from biofertilizers. Epidemiological studies have shown that cases of foodborne infections and intoxications are significantly related to animal-derived foods. The presence of these species in aquatic environments indicates areas or organisms contaminated with animal or human feces. Especially, the presence of E. coli in aquatic environments has become a serious problem worldwide. Pathogenic strains of E. coli cause waterborne and foodborne diseases. Materials and methods This study included a total of 290 samples collected from five different dairy farms between April and September 2023 which comprised 20 samples of cow manure, 20 samples of milk, three samples of dairy workers' hand washing water, five samples ofsoil, five samples of water, and five samples of vegetables. The samples taken from the farms were homogenized with 0.1% peptone water at a ratio of 1/10. They were then cultured on xylose lysine deoxycholate (XLD), eosin methylene blue agar (EMB), and blood agar media, and gram-negative colonies were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and the VITEK2 automated system (BioMerieux Inc., Durham, NC). Amplification of the isolated DNA extracts was performed with A.B.T.™ 2X HS-PCR MasterMix (A.B.T Laboratory Industry, Arnavutköy, Turkey) in the SimpliAmp™ thermal cycler (Thermo Fischer Scientific Inc., Waltham, MA) and visualized by agarose gel electrophoresis. Results Among the 52E. coli strains isolated in our study, the highest antibiotic sensitivity rate was observed in meropenem, while the lowest sensitivity rates were determined in cefazolin and cefuroxime. While two of the Salmonella spp. (n = 2) isolates were found to be resistant to tetracycline, and one was found to be resistant to penicillin and ampicillin. No resistance to trimethoprim/sulfamethoxazole was detected in either isolate. Extended-spectrum beta-lactamases (ESBLs) were detected in only four (7.7%) E. coli strains. While tetA, tetB, and TEM genes were seen in almost all E. coli strains, they were not found in Salmonella spp. Conclusion In conclusion, our study revealed the presence of antimicrobial resistance genes in E. coli and Salmonella spp. isolates collected from various farms and environmental samples, which render the antimicrobials used for disease treatment ineffective. Consequently, research should be undertaken to prevent the development of new resistance genes in our country, as creating new medications and treatment strategies for these diseases is costly and time-intensive.

Sensitivity and specificity of Nanopore sequencing for detecting carbapenem and 3rd-generation cephalosporin-resistant Enterobacteriaceae in urine samples: Real-time simulation with public antimicrobial resistance gene database

ObjectivesTo evaluate the accuracy of beta-lactamase gene detection directly from urine samples by Nanopore sequencing. MethodsDNA was extracted from bacterial pellets in spun urine. The purified DNA was then sequenced in native form by a Nanopore sequencer (MinION) to identify the organisms and beta-lactamase genes. Results were compared to routine urine cultures and standard antimicrobial susceptibility tests (AST). ResultsWe processed 60 urine samples of which routine cultures grew Enterobacteriaceae, including 28 carbapenem-resistant (CRE), 17 extended-spectrum beta-lactamase (ESBL) or AmpC producing, and 15 non-ESBL/AmpC phenotypes. We excluded 7 samples with extremely low DNA amounts (<1 ng/μl) for a final case of 53 in total. The sensitivity of antimicrobial resistance gene detection within 6 h, the optimal duration from real-time simulation, of Nanopore sequencing for the diagnosis of carbapenem-resistant and ceftriaxone-resistant phenotypes was 73.9 % (95%CI 56.0–91.9 %) and 81.1 % (95%CI 68.5–93.7 %), while the specificity was 96.7 % (95%CI 90.2–100.0 %) and 56.3 % (95%CI 31.9–80.6 %), respectively. The median times for MinION to generate DNA reads containing carbapenemase and ESBL/AmpC genes were 93 min (IQR 17–245.5) and 99 min (IQR 31.25–269.75) after sequencing commencement, respectively. ConclusionsNanopore sequencing can identify bacterial genotypic resistance in urine and may enable clinicians to adjust antimicrobial therapy earlier than routine AST.

Challenges and trends in Gram-negative bacterial infections in critically neonates: A seven-and-a-half-year observational study

Analyze the incidence, risk factors, and fatality rates of bloodstream infections by Gram-negative bacteria (GNB-BSIs) in a Neonatal Intensive Care Unit. This study employs a retrospective cohort design utilizing records of neonates admitted to the Neonatal Intensive Care Unit between January 2015 and June 2022. Among 1,495 neonates, 5.2% developed GNB-BSIs. The average incidence of infection per1,000 patient-days was 2.9. Primary risk factors for infection thatincluded preceeding carbapenem use were significant risk factors (odds ratio=514.4; P<.01) and fourth-generation cephalosporins (odds ratio=66; P<.01). Among the 85 GNB, 75.3% were fermenters, and 24.7% were non-fermenters. Of the isolates, 14.1% produced extended-spectrum beta-lactamase, and 2.3% carbapenem-resistant. Infection correlated with prolonged hospital stays (10-39days) and increased mortality (10%-29.9%). The high incidence of GNB-BSIs was exacerbated by the preceeding use of broad-spectrum antimicrobials, increasing the presence of multidrug-resistant isolates and fatality rates. These findings emphasize the importance of active surveillance.

Extended-spectrum beta-lactamase and carbapenemase producing Klebsiella Pneumoniae isolated from patient with suspected urinary tract infection

Urinary tract infection (UTI) is the most common bacterial infections in humans and serious health problem in many parts of the world. Carbapenem resistant Enterobacteriaceae (CRE) are increasingly reported from healthcare facilities in Nepal and around the world. This study is carried out with an objective to assess the Prevalence of ESBL and carbapenemase producing Klebsiella pneumoniae isolate from patient with suspected of UTI. Total 880 urine sample was collected during 6-month period and processed and identified by using conventional microbiological procedure and biochemical test. ESBL screening isolates were done by using Ceftazidime and Ceftazidime with clavulanic acid. Production of ESBL was determine by combination Disc Test. For the confirmation of carbapenemase producing K. pneumoniae Modified Hodge Method, Carbapenem Inactivation Method and EDTA Impregnated Disc test (MBL) was performed. Out of total sample, 9.30% (82) showed significant growth. Prevalence of UTI seen more in male as compare to female patients. Out of positive isolates, 37.8% were K. pneumoniae. K. pneumoniae were resistance with Ceftriaxone (54.8%) followed by 51.61% Ceftazidime, Cotrimoxazole. Multi drug resistance (MDR) was found to be 54.83%. Out of 31 isolates, 10(32%) were confirmed as ESBL positive by Combination Disc Assay. 14 isolates were screened as carbapenemase producers but 2(14.28%) showed positive through Modified Hodge Method, 4(28.57%) Modified Carbapenem Inactivation Method and 7(50%) EDTA impregnated Disc Test (MBL). Analytically, the sensitivity of MHT, CIM, and MBL tests was 42.81%, 58.10%, and 76.96%, respectively, all with 100% specificity. Colistin was the drug of choice, with 93.54% of isolates showing sensitivity to it. Early detection of ESBL and MBL-producing isolates is crucial for reducing mortality rates and preventing the intra-hospital spread of these strains.

Isolation ofaerobic bacteria from surgical site infections following orthopaedic operations incats and dogs.

Surgical site infections (SSIs) cause significant complications inthe postoperative period inveterinary surgeries. Determining the aetiology ofinfections iscrucial for increasing treatment success rates. This study aims toassess treatment processes byidentifying the bacterial agents responsible for infections occurring incats and dogs after orthopaedic operations and todetermine the antimicrobial resistance profiles ofthese agents. Strains isolated from SSIs were retrospectively analysed inpatients brought tothe Surgical Clinic ofSiirt University Animal Health Application and Research Hospital between 2021 and 2023. The isolates were identified using MALDI-TOF MS. The disc diffusion method was applied todetermine the antimicrobial susceptibility ofthe isolates. Ahigh isolation rate was detected inmethicillin-resistant Staphylococcus pseudintermedius and extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae. According tothe antimicrobial susceptibility results, cephalosporin treatment was continued inonly one ofthe cases inwhich prophylactic treatment with cephalosporin group antibiotics was applied. Consequently, this study revealed that preoperative prophylactic antibiotic administration may not besufficient inpreventing surgical site infections. Diagnosis ofaetiological agents and evaluation ofantimicrobial susceptibility are essential inpreventing surgical site infections and determining effective treatment options.

Typical pneumonia among human immunodeficiency virus-infected patients in public hospitals in southern Ethiopia.

Typical pneumonia is a pressing issue in the treatment of human immunodeficiency virus (HIV) patients, especially in Sub-Saharan Africa, where it remains a significant menace. Addressing this problem is crucial in improving health outcomes and the reduction of the burden of diseases in this vulnerable category of patients. To determine the prevalence of community-acquired typical pneumonia among HIV patients in Public Hospitals in southern Ethiopia. A cross-sectional study was done among 386 HIV patients clinically suspected of typical pneumonia attending the anti-retroviral therapy (ART) clinics of two hospitals from March to September 2022. A pretested structured questionnaire was employed to collect the demographic, clinical, and behavioral data. Sputum samples were collected and inspected for bacteria following standard procedures, and antimicrobial susceptibility testing was performed employing the Kirby-Bauer disk diffusion method. Besides, extended-spectrum β-lactamase (ESβL) and carbapenemase-producing Gram-negative bacteria were inspected by the double disk synergy test and modified carbapenem inactivation method. Descriptive and inferential statistical analyses were also done. Overall, 39.1% (151/386) of sputum cultures (95% Confidence Interval: 32.4-44) were bacteriologically positive. A total of 151 bacteria were identified, comprising 72.8% (n = 110) of Gram-negative bacteria. The predominant isolate was Klebsiella pneumoniae (25.8%, n = 39), followed by Staphylococcus aureus (17.9%, n = 27); 59.6% (n = 90) of the entire isolates were multidrug-resistant (MDR). Forty percent (11/27) of S. aureus were methicillin-resistant S. aureus (MRSA), and 28.1% (n = 31) and 20.9% (n = 23) of Gram-negative bacteria were extended-spectrum beta-lactamases (ESBL) and carbapenemase producers, respectively. Occupational status, alcohol consumption, cluster of differentiation4 (CD4) Thymocyte cell count < 350, interruption of trimethoprim-sulfamethoxazole prophylaxis and antiretroviral treatment, and recent viral load ≥ 150 were found statistically significant. The higher rates of MDR, MRSA, ESBL, and carbapenem-resistant Enterobacterales (CRE) indicate that bacterial pneumonia is a vexing problem among HIV patients and therefore it is advisable to implement an antimicrobial stewardship program in the study area.

Acute Respiratory Distress Syndrome in a Patient with Myasthenia Gravis and Septic Shock: A Case Report

Introduction: Myasthenia gravis (MG) is an autoimmune disorder characterized by muscle weakness, often affecting respiratory and oropharyngeal muscles. This predisposition to respiratory compromise, coupled with impaired swallowing and the potential need for immunosuppressive therapies, increases the risk of pneumonia and subsequent sepsis in MG patients. Sepsis, in turn, is a significant risk factor for acute respiratory distress syndrome (ARDS), a severe lung condition with high mortality. Case presentation: We present the case of a 47-year-old male with a 4-year history of MG who was admitted to our hospital with progressive dyspnea and dysphagia. His condition deteriorated rapidly, leading to septic shock and respiratory failure necessitating invasive mechanical ventilation. Blood cultures identified Klebsiella pneumoniae with extended-spectrum beta-lactamase (ESBL) production. Despite aggressive treatment, including therapeutic plasma exchange (TPE), the patient's hospital course was complicated. Conclusion: This case underscores the critical importance of vigilant monitoring and early intervention in MG patients presenting with respiratory symptoms or signs of infection. Prompt recognition and aggressive management of sepsis are crucial to mitigate the risk of ARDS and improve outcomes in this vulnerable patient population.

Distribution of ESBL-producing and non-ESBL-producing Klebsiella pneumoniae isolated from sputum specimens in the Zainoel Abidin General Hospital, Banda Aceh, Indonesia

Abstract. Suhartono S, Hayati Z, Mahdani W, Andini F. 2024. Distribution of ESBL-Producing and Non-ESBL-Producing Klebsiella pneumoniae isolated from sputum specimens in the Zainoel Abidin General Hospital, Banda Aceh, Indonesia. Biodiversitas 25: 3247-3254. Klebsiella pneumoniae, a significant nosocomial pathogen, particularly in the case of strains producing extended-spectrum beta-lactamase (ESBL), was the focus of this study. The study aimed to determine the distributions stratified by patients’ age, sex, hospital wards, and the antimicrobial susceptibility pattern of ESBL and non-ESBL K. pneumoniae isolated from expectorate sputum at Zainoel Abidin General Hospital (RSUD dr. Zainoel Abidin), Banda Aceh, Indonesia, from January 2021 to January 2024. The prevalence of ESBL-producing K. pneumoniae was 79.84% (103/129 isolates), with the highest incidence found in patients aged &gt;45 years (74.76%) and males (67.96%). The prevalence of non-ESBL-producing K. pneumoniae was 20.16% (26 isolates), with the highest incidence in patients aged &gt;45 years (76.92%) and males (62.23%). The surgical ward had the highest incidence of ESBL-producing K. pneumoniae (14.56%), followed by the cardiopulmonary ward (14.56%). The cardiopulmonary ward also had the highest incidence of non-ESBL-producing K. pneumoniae (38.47%). The ESBL-producing K. pneumoniae isolates were more sensitive to amikacin than non-ESBL isolates, while the non-ESBL-producing K. pneumoniae isolates were sensitive to cefotaxime, amikacin, and tobramycin. The increasing prevalence of ESBL-producing K. pneumoniae underscores the significance of the study's findings, providing valuable insights into the epidemiology and antibiotic susceptibility profiles of K. pneumoniae in this clinical setting and the importance of reasonable antibiotic use.

Prevalence and characterization of quinolone resistance and integrons in clinical Gram-negative isolates from Gaza strip, Palestine.

Gram-negative bacteria with quinolone resistance and extended-spectrum beta-lactamases (ESBLs) present significant treatment challenges. This study evaluated the prevalence and characteristics of quinolone resistance in Gram-negative strains, investigating the relationship between plasmid-mediated quinolone resistance (PMQR), ESBLs, and integrons. We collected 146 Gram-negative isolates from patients in three Palestinian hospitals. For quinolone resistance isolates, the presence and characterization of PMQR, β-lactamase genes and integrons were studied by PCR and sequencing. Out of 146 clinical isolates, 64 (43.8%) were resistant to quinolones, with 62 (97%) being multidrug-resistant (MDR) and 33 (51.5%) ESBL-producers. PMQR-encoding genes were present in 45 (70.3%) isolates, including aac(6')-Ib-cr (26.6%), qnrA (18.8%), qnrS1 (20.8%), and qnrB (6.4%). BlaCTX-M genes were detected in 50% (32/64) of isolates, with blaCTX-M-15 being the most common. BlaTEM-1, blaSHV-1 and blaVIM genes were found in 13, 6, and 4 isolates, respectively. Class I integrons were found in 31/64 (48%) of isolates, with 14 containing gene cassettes conferring resistance to trimethoprim (dhfr17, dfrA12, dfrA1) and aminoglycosides resistance genes (aadA1, aadA2, aadA5, and aadA6). This study found a high rate of quinolone resistance, ESBL and integrons in clinical Gram-negative isolates from our hospitals. Urgent measures are crucial, including implementing an antimicrobial resistance surveillance system, to control and continuously monitor the development of antimicrobial resistance.

A Pilot Analysis for a Multicentric, Retrospective Study on Biodiversity and Difficult-to-Treat Pathogens in Burn Centers across the United States (MICROBE).

Following burn injury, patients are at increased risk of infection and are often cited as having a high incidence of difficult-to-treat pathogens (DTp). The purpose of this study is to determine the incidence of DTp after burn injury, which factors are associated with their development, and subsequent outcomes. This single-center, retrospective study assessed patients with thermal or inhalation injury who had a positive culture resulting in initiation of treatment (i.e., excision, topical, or systemic antimicrobials). Demographic data, pathogen and resistance profiles, and prior exposure to topical and systemic antimicrobials were collected. Pathogens were considered DTp if they were multi-drug-resistant (MDR), extensively drug-resistant (XDR), methicillin-resistant Staphylococcus aureus (MRSA), extended-spectrum beta-lactamase (ESBL)-producing, AmpC-producing, carbapenem-resistant, difficult-to-treat resistance (DTR) Pseudomonas sp., carbapenem-resistant Acinetobacter baumannii (CRAB), or Stenotrophomonas spp. Sixty-five patients who grew 376 pathogens were included in the final analysis. Two-hundred thirteen (56.7%) pathogens were considered DTp. Prior exposure to 7 of the 11 collected topical antimicrobials and 9 of 11 systemic antimicrobial classes were significantly associated with future development of a DTp. This remained true for six and eight, respectively, after controlling for significant covariates via logistic regression. As there were only four deaths, a Cox-proportional hazard analysis was not feasible. The Kaplan-Meier plot according to DTp revealed a clear divergence in mortality (Log rank p = 0.0583). In this analysis, exposure to topical and systemic antibiotics was associated with the development of DTp. The results from this pilot study will inform the next iteration of multicenter study.

Effects of household concrete floors on maternal and child health - the CRADLE trial: a randomised controlled trial protocol.

Early life soil-transmitted helminth infection and diarrhea are associated with growth faltering, anemia, impaired child development, and mortality. Exposure to fecally contaminated soil inside the home may be a key contributor to enteric infections, and a large fraction of rural homes in low-income countries have soil floors. The objective of this study is to measure the effect of installing concrete floors in homes with soil floors on child soil-transmitted helminth infection and other maternal and child health outcomes in rural Bangladesh. The Cement-based flooRs AnD chiLd hEalth (CRADLE) trial is an individually randomised trial in Sirajganj and Tangail districts, Bangladesh. Households with a pregnant woman, a soil floor, walls that are not made of mud will be eligible, and no plan to relocate for 3 years. We will randomise 800 households to intervention or control (1:1) within geographic blocks of 10 households to account for strong geographic clustering of enteric infection. Laboratory staff and data analysts will be blinded; participants will be unblinded. We will install concrete floors when the birth cohort is in utero and measure outcomes at child ages 3, 6, 12, 18, and 24 months. The primary outcome is prevalence of any soil-transmitted helminth infection (Ascaris lumbricoides, Necator americanus, or Trichuris trichiura) detected by qPCR at 6, 12, 18, or 24 months follow-up in the birth cohort. Secondary outcomes include household floor and child hand contamination with E. coli, extended-spectrum beta-lactamase producing E. coli, and soil-transmitted helminth DNA; child diarrhea, growth, and cognitive development; and maternal stress and depression. Study protocols have been approved by institutional review boards at Stanford University and the International Centre for Diarrheal Disease Research, Bangladesh (icddr,b). We will report findings on ClinicalTrials.gov, in peer-reviewed publications, and in stakeholder workshops in Bangladesh. NCT05372068, pre-results.