ESBL Producers Research Articles

THE CONTINUING MENACE OF EXTENDED SPECTRUM BETA LACTAMASE (ESBLS) - A CENTRE BASED STUDY

Abstract:
 Objective: To determine the occurrence of Extended-spectrum β-lactamase (ESBL) producing gram negative bacteria (GNB) and their sensitivity patterns in a tertiary care hospital.
 Methods: In this cross-sectional study, conducted in the Microbiology section of Pathology lab, in a tertiary care hospital Lahore, various specimens such as blood, body fluids, urine, and pus were included during a period starting from October 2020 till September 2021. The samples having growth of gram negative bacteria were processed further for determination of ESBL production. After isolation, antimicrobial susceptibility pattern was determined for each isolate.
 Results: GNB were isolated from 130 samples, among those 46 (35.4%) were labelled as ESBL. The commonest ESBL producing bacteria was E. coli, followed by pseudomonas aeruginosa found in 17.39% samples. In E. Coli, Imipenem (IPM) showed highest sensitivity of 84.8%, other sensitive drugs were Amikacin (AK), gentamicin (CN), and Tazobactam-piperacillin(TZP). For Pseudomonas aeruginosa, Imipenem and Tazobactam-piperacillin showed sensitivity of 75%, AK and CN were 50% sensitive. Against Klebsiella pneumoniae , IPM was 100% sensitive, AK and TZP were 85.7% sensitive. IPM, CN, and AK were 100% sensitive against Morganella morganii. AK and CN were 100% sensitive against Citrobacter species.
 Conclusion: It was concluded that, there was significant prevalence of ESBL producing strains amongst Gram negative bacterial infections. E. Coli and Pseudomonas aeruginosa were the commonest ESBL producers. IPM, AK, and TZP were sensitive against majority of the ESBL producing GNB.

Antibiotic resistance patterns in nosocomial infections: preliminary data from Hospital of Al-Hoceima, Morocco.

Nosocomial infections or healthcare-associated infections (HCAIs) represent a public health problem. The burden of antibiotic resistance in these infections is still unclear in Morocco. The objective of this study was to describe antibiotic susceptibility of the main bacteria responsible for nosocomial infections in order to propose prevention measures. Data were collected from 1519 laboratory records including hospital inpatients suspected of nosocomial infections in Mohamed V Hospital of Al-Hoceima between January 2016 and December 2020. The data were analyzed using SPSS software version 25. Bacteriological test samples included 65.5% of urine, 27.2% of pleural fluid, 4.5% of pus, and 2.8% of protected distal swab. Two hundred and twenty-seven (15%) samples were culture-positive. The bacteria isolated were mainly enterobacteria (Escherichia coli, 43.6% and Klebsiella pneumoniae, 13%), non-fermentative Gram-negative bacteria (Pseudomonas aeruginosa, 10.8%), and Staphylococcus aureus (24.3%). Extended spectrum beta-lactamases (ESBLs)-producing Enterobacteriaceae represented 25.4% and those resistant to other families of antibiotics accounted for 12.5%. In our study, we reported 17% ESBL producers among urinary infections. Methicillin-resistant Staphylococcus aureus accounted for 22.2%. Pseudomonas aeruginosa that were resistant to ticarcillin, ceftazidime, and imipenem represented 29% of the cases. Our results showed a higher frequency of resistance. A microbiological surveillance system is highly needed to identify bacterial niches in the hospital environment at Mohamed V Hospital.

Phenotypic detection of extended-spectrum beta-lactamase produced by Klebsiella pneumoniae isolated from different clinical samples from intensive care unit, Khamis Military Hospital, Abha, Kingdom of Saudi Arabia

Background: Klebsiella pneumoniae is an opportunistic pathogen that is one of the commonest causes of infection in intensive care units. K. pneumoniae infection have contributed considerably to morbidity and mortality in patients with chronic illnesses. The coexistence of virulence factors and the genetic determinant of antibiotic resistance complicates treatment outcomes. The emergence of pathogenic multi-drug resistant K. pneumoniae poses a great threat to the healthcare system. Thus, the aim of this study was to isolate and to phenotypically detect the extended-spectrum beta-lactamase produced by K. pneumoniae that are isolated from different clinical samples from ICU in Khamis Military Hospital. Methods: A retrospective observational study was conducted at Khamis Military Hospital from December 2021 to April 2022. Clinical samples collected within the ICU underwent microbiological identification phase to confirm the presence of K. pneumoniae. This was followed by antibiotic susceptibility testing to various antimicrobial agents to detect the production of carbapenemases (KPC, NDM-1, OXA-48) and extended spectrum beta lactamase (ESBL). The collected data were subjected to statistical analysis using appropriate tools. Descriptive statistics were used to summarize the demographic and clinical characteristics of the patient population from whom the samples were obtained. Results: A total of 114 isolates of K. pneumoniae were identified from different clinical samples of ICU patients. The percentage of ESBL production constituted 28.94% (33 isolates). Regarding the distribution of carbapenemases production, 43 isolates (37.7%) had the KPC and NDM carbapenemases, while 34 isolates (29.8%) had the OXA-48 carbapenemases. Conclusions: The high prevalence of K. pneumoniae represents a threat to ICU patients, thus, this issue should be treated with urgent attention. Our data informs the need for regular surveillance of antibiotic resistance in pathogenic bacteria in clinical settings for meaningful control of emergence and spreading of multi-drug-resistant K. pneumoniae.

Impact of Solidago virgaurea Extract on Biofilm Formation for ESBL-Pseudomonas aeruginosa: An In Vitro Model Study.

The increasing disparity between antimicrobial resistance (AMR) and the development of new antimicrobials continues to pose a significant global health concern. However, plant extracts have shown promise in combating this issue either through their inherent antimicrobial activity or by serving as potential reservoirs of effective antimicrobial compounds. These compounds have the ability to target pathogenic biofilms and inhibit the production of extended-spectrum β -lactamases (ESBLs). However, there is limited research available on the antibacterial properties of goldenrod extract. Thus, the objective of this study was to investigate the impact of S. virgaurea (SV) extract on the viability and ability to form biofilms of ESBL-Pseudomonas aeruginosa (P. aeruginosa). A cross-sectional study was conducted from August 2022 to March 2023. The broth microdilution method was employed to determine the minimum inhibitory concentration (MIC) of the (SV) extract. Subsequently, the minimum bactericidal concentration (MBC) was determined based on the MIC values obtained. The antibiotic susceptibility of bacteria was evaluated using the Kirby disk diffusion assay and an Antimicrobial Susceptibility Testing (AST) card in conjunction with the Vitek-2 compact system. Biofilm formation was evaluated using Congo red and a 96-well Elisa plate, while the presence of extended-spectrum β-lactamases (ESBLs) was estimated by measuring the reduction of nitrocefin at a wavelength of 390 nm. In addition, treatment of biofilm and ESBL activity with SV extract using 96-well Elisa plate and nitrocefin hydrolyzing, respectively. The resistance rates of P. aeruginosa isolates to the tested antibiotics were as follows: Levofloxacin 33%, Ciprofloxacin 40%, Amikacin 49%, Meropenem 50%, Cefepime 70%, Ceftazidime 75%, Cefotaxime 85%, Piperacillin-Tazobactam 90%, Amoxiclav 97%, Ampicillin 99%, Ceftriaxone 100%. The prevalence of MDR-P. aeruginosa, XDR-P. aeruginosa, PDR-P. aeruginosa and non-MDR-PA were 40% (n = 40), 7% (n = 7), 3% (n = 3) and 50% (n = 50), respectively. From the GC-MS results, it was observed that the presence of Octadecane, Clioquinol, Glycerol tricaprylate, hexadecanoic acid, cis-13-octadecenoic acid, oleic acid and Propanamide were the major components in the Solidago extract. In the determination of plant crude extracts, the values ranged between 0.25 and 64 mg/mL against bacteria. The resulting activity of the extract showed a significant statistical relationship at a p-value ≤ 0.01 against ESBL production and biofilm formation in P. aeruginosa. The S. virgaurea extract exhibited effectiveness in inhibiting biofilm formation and combating P. aeruginosa strains that produce extended-spectrum β-lactamases (ESBLs).

Prevalence and characterization of ESBL-producing Escherichia coli in healthy pregnant women and hospital environments in Benin: an approach based on Tricycle.

Extended-Spectrum Beta-Lactamase (ESBL)-producing Enterobacterales are recognized as significant pathogens due to their resistance to multiple antibiotics. This study aimed to determine the prevalence of ESBL-producing Escherichia coli (E. coli) in different settings, including healthy pregnant women, the food chain, and the environment of tertiary hospitals in Benin. Samples were collected from various sources, including fecal samples from healthy pregnant women, food samples from hospital canteens, and hospital effluents from four tertiary hospitals in southern Benin. Fecal samples were plated on MacConkey agar supplemented with cefotaxime (4 μg/mL), while food and water samples were plated on Tryptone Bile X agar supplemented with cefotaxime (4 μg/mL). Urea indole tests were used for preliminary identification of E. coli colonies, followed by confirmation of ESBL production using the double disk synergy technique. Antibiotic susceptibility testing of ESBL-producing E. coli strains was conducted using the disk diffusion method on MH agar. Polymerase Chain Reaction (PCR) was used to investigate the presence of ESBL-encoding genes. Among the 296 fecal samples collected from four tertiary hospitals, ESBL-producing E. coli was isolated from 22.30% (66) of the samples. All E. coli isolates from hospital effluents exhibited ESBL production, while ESBL-producing E. coli was not detected in food and drinking water samples. The analysis of variable associations showed no significant associations (p > 0.05) for the studied factors. Antibiotic susceptibility testing revealed high resistance rates among the ESBL-Ec isolates against several tested antibiotics, including amoxicillin, aztreonam, ceftriaxone, ciprofloxacin, and trimethoprim-sulfamethoxazole. However, most isolates remained susceptible to ertapenem, amoxicillin-clavulanate, and imipenem. The most prevalent ESBL-encoding genes were blaTEM (37.50%), blaOXA-1 (19.44%), and blaSHV (11.11%), while a smaller proportion of isolates carried blaCTXM-1/blaCTXM-15 (5.55%) and blaCTXM-9. This study provides insights into the prevalence of ESBL-producing E. coli carriage in the feces of healthy pregnant women in southern Benin. Additionally, it highlights hospital wastewater as a potential reservoir of ESBL-producing bacteria in the environment. The detection of ESBL-producing E. coli in hospital effluents raises concerns about the dissemination of antibiotic resistance genes into the environment. The high resistance rates observed among ESBL-Ec isolates against commonly used antibiotics emphasize the urgent need for antimicrobial stewardship and infection control measures. The identification of prevalent ESBL-encoding genes contributes to understanding the genetic basis of ESBL resistance in the studied population. Further research is warranted to explore the mechanisms of transmission and potential interventions to mitigate the spread of ESBL-producing Enterobacterales.

Antimicrobial Resistance Pattern and Empirical Antibiotic Treatments in Neonatal Sepsis: A Retrospective, Single-Center, 12-Year Study.

Neonatal sepsis is an important cause of morbidity and mortality in neonatal intensive care units (NICUs). Continuous evaluation of antimicrobial resistance (AMR) profiles is advised to implement antimicrobial stewardship (AMS) programs and establish effective empiric antibiotic protocols. AMS may reduce AMR in NICUs and improve sepsis outcomes. In this retrospective observational study, we report data on culture-positive neonatal sepsis, assessing differences after the implementation of an AMS program (2011-2016 vs. 2017-2022). A total of 215 positive bacterial cultures from 169 infants were retrieved, with 79 early-onset (36.7%) and 136 late-onset (63.3%) sepsis episodes. Frequent causative agents for early-onset sepsis were S. agalactiae and E. coli, all susceptible to empiric treatment. Late-onset sepsis was mainly caused by Enterobacterales and S. aureus. Aminoglycosides, cefotaxime, and piperacillin-tazobactam resistance among Enterobacterales was substantially low; S. aureus was mostly susceptible to oxacillin and vancomycin. There were no differences in mortality and multidrug-resistant pathogens rates between the two study periods. There were five episodes of fungal late-onset sepsis, mostly due to C. albicans, of which one was fatal. The microbial distribution pattern and AMR profiles overlapped with other European studies. Because susceptibility patterns are rapidly changing worldwide, with the emerging threat of Methicillin-resistant S. aureus and extended-spectrum beta-lactamases producers, infection prevention and control practices and AMS strategies require continuous optimization to limit selection pressure and AMR escalation.

Beta-lactam Resistance Profile among Klebsiella pneumoniae Clinical Isolates from Alexandria, Egypt.

Klebsiella pneumoniae is a major drug-resistant human pathogen accountable for a wide range of infections. In this cross-sectional study, we aimed to determine the phenotypic and genotypic features of β-lactamase-producing K.pneumoniae clinical isolates from Alexandria, Egypt. A total of 50 nonduplicated clinical isolates of K.pneumoniae were obtained from various specimens. They were identified biochemically and by biotyping using mass spectrometry. For molecular characterization, plasmid profile analysis was performed. Screening for extended spectrum β-lactamases (ESBLs), carbapenemases and AmpC production was carried out phenotypically and genotypically. Correlation analysis was performed to assess the relationship between phenotype, genotype and resistance patterns among the studied isolates. The dendrogram demonstrated 38 distinct plasmid profiles among 62% of our isolates. According to antimicrobial susceptibility testing, 90% of isolates were multi/extensive-drug resistant. Nineteen out of 50 (38%) were resistant to cefoxitin, while only 10 (20%) were resistant to imipenem. All isolates were susceptible to colistin. Phenotypically, ESBL producers (78%) were the most common, followed by carbapenemase producers (24%). Genotypically, the most common ESBL gene was blaSHV (90%), followed by blaCTX-Mu (74%), while the most common carbapenemase genes were blaNDM (56%) and blaOXA-48 (54%). No blaKPC or blaIMP were detected. Plasmid-mediated AmpC resistance was confirmed in only two out of 19 cefoxitin-resistant isolates. Both the blaNDM and blaOXA.48 genes were significantly positive correlated (rho = 0.56, p = 0.004). Absence of blaKPC among carbapenem resistant K.pneumoniae isolates in Alexandria, Egypt. AmpC production is not the main factor behind the resistance to cefoxitin among our isolates.

Phenotypic detection of extended spectrum beta-lactamase production in clinical and non - clinical isolates of klebsiella species of animal origins in Abeokuta, Nigeria.

Production of extended-spectrum β-lactamases (ESBLs) confer resistance to β-lactam antibiotics. This study investigated the presence of ESBL-producing Klebsiella spp in clinical and non-clinical samples from different animal species in Abeokuta, Nigeria. The species of Klebsiella were determined by biochemical characterization (Oxoid Microbact GNB 24E®), while phenotypic ESBL-production was confirmed by using the cefpodoxime and cefpodoxime/clavulanic acid combination disc kit. Ninety-five Klebsiella isolates were investigated in this study. Fifty-five (57.9%) of the isolates were obtained from the faeces of apparently healthy animals while 40 (42.1%) were from clinical samples. The Klebsiella isolates were identified as follows: K. oxytoca (34.7%), K. pneumoniae (26.3%), K. ozaenae (18.9%), K. terrigena (13.7%), K. rhinooscleromatis (5.3%), K. planticola (1.1%). Eight (8.4%) out of the 95 isolates of Klebsiella spp were identified as ESBL-producers. These included four isolates of Klebsiella pneumoniae, three of Klebsiella oxytoca and one of Klebsiella ozaenae. Five out of the eight ESBL-producing isolates were from clinical samples while three isolates were from the faeces of apparently healthy animals. The ESBL-producing isolates were 100% resistant to ampicillin, cefotaxime, ceftazidime, cefpodoxime, streptpmycin, tetracycline, compound sulfonamide, trimethoprim/sulphomethoxazole, and nalidixic acid. Apparently healthy carriers and sick animals can serve as sources of transmission of ESBL-producing Klebsiella spp to other animals and humans.

In Vitro Activity of New β-Lactamase Inhibitor Combinations against blaNDM, blaKPC, and ESBL-Producing Enterobacteriales Uropathogens.

Antibiotic resistance in uropathogens has increased substantially and severely affected treatment of urinary tract infections (UTIs). Lately, some new formulations, including meropenem/vaborbactam (MEV), ceftazidime/avibactam (CZA), and ceftolozane/tazobactam (C/T) have been introduced to treat infections caused by drug-resistant pathogens. This study was designed to screen Enterobacteriales isolates from UTI patients and to assess their antimicrobial resistance pattern, particularly against the mentioned (new) antibiotics. Phenotypic screening of extended-spectrum β-lactamase (ESBL) and carbapenem resistance was followed by inhibitor-based assays to detect K. pneumoniae carbapenemase (KPC), metallo-β-lactamase (MBL), and class D oxacillinases (OXA). Among 289 Enterobacteriales, E. coli (66.4%) was the most predominant pathogen, followed by K. pneumoniae (13.8%) and P. mirabilis (8.3%). The isolates showed higher resistance to penicillins and cephalosporins (70-87%) than to non-β-lactam antimicrobials (33.2-41.5%). NDM production was a common feature among carbapenem-resistant (CR) isolates, followed by KPC and OXA. ESBL producers were susceptible to the tested new antibiotics, but NDM-positive isolates appeared resistant to these combinations. KPC-producers showed resistance to only C/T. ESBLs and carbapenemase encoding genes were located on plasmids and most of the genes were successfully transferred to recipient cells. This study revealed that MEV and CZA had significant activity against ESBL and KPC producers.

Oral fosfomycin for treatment of acute bacterial prostatitis caused by multidrug-resistant Enterobacterales.

To assess the feasibility of oral fosfomycin-tromethamine (FT) for the management of acute bacterial prostatitis (ABP) caused by multidrug-resistant (MDR) Enterobacterales. An observational study of adult patients diagnosed with ABP from Vall d'Hebron University Hospital (Barcelona, Spain), treated with oral FT. The primary outcome was clinical cure defined as symptom relief at the control visit, 2-4 weeks post-end of treatment. Secondary outcomes included microbiological cure, relapse, and adverse events related to the treatment. Eighteen patients with ABP caused by Enterobacterales (15 Escherichia coli and three Klebsiella pneumoniae) were included. Microorganisms were MDR bacteria [14 extended-spectrum beta-lactamase (ESBL) producers and two carbapenemase producing K. pneumoniae]. Patients received treatment with FT 3 g/48 hours during a median of 14 days (Q25-Q75, 12-17.75). Fifteen patients received a lead-in phase of intravenous suitable antimicrobial during a median of 7 days (Q25-Q75, 3.75-8). No patient had to stop treatment due to adverse events, and the only side effect reported in two patients was diarrhea. Clinical cure was achieved in all (18/18) patients and microbiological cure in 11/12 patients. After a median of follow-up of 5 months (Q25-Q75, 2-11), 2/18 patients relapsed with an orchitis and a new episode of ABP. FT is an attractive step-down therapy for ABP in patients with resistance or side effects to first-line drugs. The availability of oral treatment could reduce the use of the carbapenems, with a benefit in the quality of life of the patient, health costs, and an ecological impact. IMPORTANCE We present a brief but largest and interesting experience in which we use fosfomycin-tromethamine (FT) for the treatment of acute bacterial prostatitis (ABP) due to multiresistant bacteria. Our study provides new data that help to consider FT as a plausible alternative for treating ABP in patients with resistance or side effects to first-line drugs. The availability of an alternative oral treatment to avoid the use of the carbapenems could have important benefits in terms of quality of life of the patient, health costs, and an ecological impact.

Acinetobacter baumannii producing ESBLs and carbapenemases in the Intensive Care Units developing fosfomycin and colistin resistance

Acinetobacter baumannii is responsible for causing difficult-to-treat healthcare-associated infections globally, owing to its resistance to antibiotics. The intensive care unit (ICU) settings mediate spread of multidrug resistance (MDR) strains. This research aimed to evaluate non-susceptible colistin and fosfomycin A. baumannii, harboring extended-spectrum beta-lactamases (ESBLs) and carbapenemases in ICU setting. During the period of 2019-2021, this study obtained 200 A. baumanni isolates out of 1410 burns samples from an ICU setting. The antibiotic sensitivity, ESBLs and carbapenemase production were determined using clinical and laboratory standards institute (CLSI) 2020. The colistin (mcr-1 and mcr-2) and fosfomycin (fosA3) resistance genes was amplified. The highest resistance was to ceftazidime (98%), cefepime (86%), tetracycline (84%), levofloxacin (78%) and piperacillin-tazobactam (76%), while the highest sensitivity was to meropenem (63%) and tigecycline (62%). ESBL production was determined in 94% and carbapenemases were observed in 54% of A. baumannii. Four isolates (2%) were found to carry the mcr-1 gene, and three isolates (1.5%) were found to carry the mcr-2 gene. Moreover, the fosA3 was not detected in the isolates. This study showed that MDR A. baumannii was high in ICU settings. The spread of antibiotics considered the last line of defense against infections is a concern that necessitates surveillance and control measures.

Antimicrobial resistance among common clinical isolates from Wayanad district

: The emerging multi-drug resistant variants in different clinical isolates is leading to increased morbidity and mortality, due failure in treatment. The paucity of an accurate data of antimicrobial resistance from different geographical areas is a major setback to its control and management. The aim of this study was to analyse the occurrence of drug resistant organisms from different clinical samples in the district of Wayanad, and also to determine the most prevalent and emerging bacterial pathogens among them. : A seven-month retrospective study of different bacterial isolates from various clinical samples was conducted in a tertiary care hospital in Wayanad district. Clinical samples taken for the study included urine, pus, sputum and blood. : Data from 2125 clinical samples were studied, in which 661 were urine samples, 910 were pus samples, 225 were blood samples and 339 were sputum samples. The predominant bacteria identified from urine sample was among which the prevalence of extended spectrum beta lactamase (ESBL) producing was 40.61%, and the metalobetalactamase (MBL) producers 3.45%. was the predominant bacteria in the pus samples, in which Methicillin resistant (MRSA) was found to be 3.92%. was the most predominant bacteria in the blood samples, in which ESBL producing was noted as 3.57%. Klebsiella species were the predominant bacteria in the sputum samples, in which ESBL producing was 16.79% and MBL producers were 3.82%.: The study helped to identify the most predominant antibiotic resistant strains from each of the clinical samples in a resource limited setting like Wayanad. Similar studies would help in successfully formulating treatment strategies against bacterial infections, thereby reducing morbidity and mortality in patients.

Fecal carriage of extended spectrum beta-lactamase and fluoroquinolone resistant gene in non-typhoidal Salmonella enterica isolates from food-producing animals and humans

This study seeks to determine the feacal carriage of extended spectrum beta-lactamase and fluoroquinolone resistant non-typhoidal Salmonella enterica isolates from food-producing animals and humans. A total of three hundred (300) fecal samples were collected using sterile universal containers from food-producing animals namely (Chicken [100], Pig [100] and humans (100) from Onicha Local Government Area of Ebonyi State and analyzed for the presence of non-typhoidal Salmonella enterica using standard microbiological techniques. Phenotypic detection of extended-spectrum beta-lactamase (ESBL) were done by disc diffusion and Double Disk Synergy Test. Molecular characterization for ESBL and fluoroquinolone-resistant genes were done by PCR with specific primers. The result shows that non-typhoidal Salmonella species (NTS) accounted for 25 % and 17 % in poultry and pig fecal sample respectively while 60 % and 40% were phenotypic ESBL producers respectively. When compared statistically there is significant difference among isolates confirmed ESBL-positive (P˂ 0.05). Also, none of the 16 (58 %) NTS isolated from humans harbored ESBL phenotype. PCR analysis with β-lactam specific primer detected the presence of blaOXA 50 % and 50 %, blaSHV 36 %, and 64 %, blaTEM 43 % and 57 %, blaCTX-M 36 % and 64 % in poultry and pig respectively. Fluoroquinolone resistant gene QnrA was present in 0 and 100 % of poultry and pig respectively. QnrB was 40 % and 60 % in poultry and pig isolates respectively. QnrS was present in 64 % isolates of poultry and 13 % isolates in pig. The high prevalence of genes encoding beta-lactamases and fluoroquinolone resistance (TEM, SHV, CTX-M and OXA, (qnrA, qnrB and qnrS) were present more in poultry and pig than in humans and demonstrate a significant public health threat from consumption of food-producing animal harboring such pathogenic resistant genotype if not properly controlled.
 Keywords: Extended spectrum beta-lactamase, Fluoroquinolone, non-typhoidal Salmonella enterica, feacal carriage

Analyses of Extended-Spectrum-β-Lactamase, Metallo-β-Lactamase, and AmpC-β-Lactamase Producing Enterobacteriaceae from the Dairy Value Chain in India.

The consumption of milk contaminated with antibiotic-resistant bacteria poses a significant health threat to humans. This study aimed to investigate the prevalence of Enterobacteriaceae producing β-lactamases (ESBL, MBL, and AmpC) in cow and buffalo milk samples from two Indian states, Haryana and Assam. A total of 401 milk samples were collected from dairy farmers and vendors in the specified districts. Microbiological assays, antibiotic susceptibility testing, and PCR-based genotyping were employed to analyze 421 Gram-negative bacterial isolates. The overall prevalence of β-lactamase genes was 10% (confidence interval (CI) (7-13)), with higher rates in Haryana (13%, CI (9-19)) compared to Assam (7%, CI (4-11)). The identified β-lactamase genes in isolates were blaCMY, blaMOX, blaFOX, blaEBC, and blaDHA, associated with AmpC production. Additionally, blaCTX-M1, blaSHV, and blaTEM were detected as ESBL producers, while blaVIM, blaIMP, blaSPM, blaSIM, and blaGIM were identified as MBL producers. Notably, Shigella spp. were the dominant β-lactamase producers among identified Enterobacteriaceae. This study highlights the presence of various prevalent β-lactamase genes in milk isolates, indicating the potential risk of antimicrobial-resistant bacteria in dairy products. The presence of β-lactam resistance raises concern as this could restrict antibiotic options for treatment. The discordance between genotypic and phenotypic methods emphasizes the necessity for comprehensive approaches that integrate both techniques to accurately assess antibiotic resistance. Urgent collaborative action incorporating rational and regulated use of antibiotics across the dairy value chain is required to address the global challenge of β-lactam resistance.

High prevalence of Mucosa-Associated extended-spectrum β-Lactamase-producing Escherichia coli and Klebsiella pneumoniae among Iranain patients with inflammatory bowel disease (IBD)

BackgroundSeveral pieces of evidence suggest that certain pathobionts belonging to Enterobacterales are associated with the development and progression of inflammatory bowel diseases (IBD). Extended-spectrum β-lactamases (ESBLs) ESBLs are frequently found in the Enterobacterales members, particularly in Escherichia coli and Klebsiella spp., and might trigger antibiotic-induced perturbations of the intestinal microbiota and led to more severe disease activity in IBD. Therefore, the severity of IBD could be influenced by ESBL-producing Enterobacterales, and hence, this study aimed to investigate the presence of ESBLs and carbapenemases among mucosa-associated E. coli and Klebsiella pneumoniae isolated from colonic biopsies of Iranian patients with IBD.MethodsIn this cross-sectional study, E. coli and K. pneumoniae were isolated from inflamed ileum and/or colon tissue of patients with IBD, including Ulcerative colitis (UC) and Crohn’s disease (CD), during colonoscopy. Demographic data and clinical characteristics were recorded, and UC and CD disease activity and extent were evaluated according to the full Mayo score and Crohn’s disease activity index (CDAI), respectively. Phenotypic and molecular detection of ESBL- and carbapenemase-producing E. coli and Klebsiella pneumoniae were carried out. Disease activity and other clinical and microbial features were compared in patients with and without gut colonization with ESBL producers.ResultsA total of 83 IBD patients, including 67 UC and 16 CD, were enrolled in the initial analysis. Intestinal colonization with ESBL-producing E. coli and/or Klebsiella pneumoniae was found in 37 (55.2%) of UC and 9 (56.2%) of DC patients – mostly harbored E. coli containing the blaCTX−M and blaTEM genes. UC patients with intestinal colonization with ESBL-producers had more severe disease compared with patients without colonization. Moreover, 10.2% of tested E. coli and 34.8% of K. pneumoniea were recognized as potential carbapenemase producers.ConclusionIntestinal colonization with ESBL producers could arise disease activity in IBD patients. Further large-scale case-control studies should be performed to investigate the possible confounding factors that could contribute to this outcome.

Antimicrobial Resistance and Plasmid Profiling of ESBL Producing Klebsiella spp. of Bovine Origin of Eastern Plain Zone of Uttar Pradesh

Emergence of antimicrobial resistance in animal husbandry is of utmost concern as it has resulted in therapeutic failure and nowadays it is one of the latest challenges faced by scientific community across the world. Extensive use of antibiotics without performing antibiotic susceptibility testing may lead to development of multidrug resistant (MDR) organisms. This study was undertaken to assess the AMR pattern and their correlation with plasmid profiling in ESBL producing Klebsiella spp. from bovine origin. Total 160 samples were collected from 2 district of Eastern Plain Zone of Uttar Pradesh (India) and 63 (39.37%) isolates were confirmed as Klebsiella spp. by conventional methods (cultural characteristics on selective media & biochemical tests). Antimicrobial susceptibility testing of confirmed isolates revealed 82.0%-94.0% resistance to 3rd & 4th generation cephalosporin, 100% to ampicillin and all isolates were found 100% sensitive to aminoglycosides, chloramphenicol and polypeptides class of antibiotics. Total 49 (77.8%) isolates were confirmed as ESBL producers using DDST and ESBL E-strip test, out of which 45 (71.43%) isolates revealed the presence of bla-CTX-M gp genes by PCR analysis. On plasmid profiling, total 82.82% isolates revealed the presence of plasmid with molecular weight between 2.5 kb to >10 kb and number 1-2. These high molecular weight plasmids are mostly situated on conjugative plasmid, which are horizontally transferred from one species to another in highly populous country like India.

Bacteriological Profile of Surgical Site Infection Following Gastrointestinal Surgery and Their Antibiogram.

Surgical site infection is one of the common complication following abdominal surgery. It causes great morbidity and mortality, further increasing prevalence of multidrug resistant bacteria have made its management very challenging. The current study aims to identify causative agent responsible for surgical site infection and their antibiotic resistance patterns. This study was conducted among patients developing surgical site infection following gastrointestinal surgery in Tribhuvan university teaching hospital over a period of one year. The samples were collected and processed according to standard methods. The bacterial pathogens with their antimicrobial susceptibility were determined and resistant pattern like methicillin resistant Staphylococcus aureus and extended spectrum beta lactamase were further detected. A total of 832 patients had under gone gastrointestinal surgery during the study period. Among them, 162 cases (19.5%) developed surgical site infection and 125 cases showed growth in culture. A total of 160 aerobic bacteria were isolated; Escherichia coli (29.9%) was the commonest organism with 40.8% being extended spectrum beta lactamase producer and 47.4% of Staphylococcus aureus were methicillin resistant. About 75.9% (85/112) of gram negative bacteria and 60.4% (29/48) gram positive bacteria were multi drug resistant. The burden of multi drug resistant bacteria causing surgical site infection is high which needs to be addressed timely. Good surveillance of bacterial antibiogram and rational antimicrobial use is necessary to reduce emergence and spread of resistant bacteria.

Resistome-based surveillance identifies ESKAPE pathogens as the predominant gram-negative organisms circulating in veterinary hospitals.

Healthcare-associated infections (HCAIs) associated with extended-spectrum cephalosporin-resistant gram-negative (ESC-R GN) bacteria are an emerging concern in veterinary hospitals, especially in companion animal intensive care units (ICUs). To understand the molecular epidemiology of ESC-R GN isolates in two veterinary hospitals (equine and small animal), a 6-month pilot study was performed during which fecal and environmental samples were obtained twice from selected patients, upon ICU admission and after 48 h of hospitalization. In total, 295 ESC-R GNs were analyzed using the Acuitas Resistome® Test (OpGen, Maryland, US), a PCR-based assay screening for 50 antimicrobial resistance gene families encoding for production of extended-spectrum beta-lactamase (ESBLs), TEM/SHV/OXA or AmpC beta-lactamases and carbapenemases. Combining organism identification and antimicrobial susceptibility data to genotyping results, unique "Acuitas profiles" were generated that can be used for fast typing the isolates and tracking transmission events. ESKAPE GN pathogens were the most prevalent ESC-R GN isolates circulating in both the small animal and equine hospitals, consisting of Enterobacter cloacae complex (21.7%), Pseudomonas aeruginosa (20%), Klebsiella pneumoniae (15.9%), and Acinetobacter baumannii complex (13.6%) followed by Escherichia coli (12.2%), most harboring a combination of genes encoding for beta-lactamases and ESBLs. Some ESKAPE genotypes showed likely intra-hospital transmission, including E. cloacae (two genotypes, one carrying SHV4, SHV5, and TEM7 and the other TEM1, TEM3, and TEM7 enzymes) in the equine and K. pneumoniae (SHV1, SHV5, and DHA1-positive) in the small animal ICUs, respectively. Furthermore, P. aeruginosa (carrying OXA-50), A. baumannii complex (OXA-51), and E. coli (CTX-M-1) genotypes were isolated across both hospitals, suggesting possible transfer mediated via movement of staff and students. Importantly, isolates carrying transmissible resistance to last-resort antimicrobials (i.e. carbapenems) were identified within the hospital environments, consisting of three environmental Acinetobacter spp. harboring blaOXA - 23 and one clinical E. coli with blaOXA - 48. We describe the widespread occurrence of ESKAPE gram-negative organisms in veterinary ICU patients and hospital environments. Findings from this project provide baseline data on the epidemiology of ESKAPE pathogens in veterinary settings, which can inform infection control policies to aid in patient management and prevent transmission of nosocomial infections associated with these pathogens.

Bacterial etiology, antimicrobial resistance and factors associated with community acquired pneumonia among adult hospitalized patients in Southwest Ethiopia.

Antibiotic resistance is a significant problem that restricts the options for treating bacterial pneumonia. This research aimed to determine the bacterial causes of pneumonia and antibiotic resistance among hospitalized patients in southwest Ethiopia. We collected and analyzed 150 sputum samples from individuals with community-acquired pneumonia from April 1st to October 30th, 2019. Standard bacteriological procedures were used to identify the bacteria. Kirby Bauer's disk diffusion method was used to assess the bacteria's susceptibility patterns. Production of carbapenemase and extended-spectrum-lactamase were confirmed phenotypically. Odds ratios and the chi-square test were computed. On the whole, bacterial pathogens were verified in 50% of the sputum samples. The predominant bacterial isolates were Klebsiella species, followed by Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus pneumoniae. About 77.5% of isolates were multidrug resistant. Moreover, 40.5% and 10.8% of the isolates were ESBL and carbapenemase producers, respectively. Aging, tobacco smoking, previous history of pneumonia, heart disease, and chronic respiratory disease had association with sputum culture-positivity. As a result, it is important to regularly monitor the bacterial etiologies and their patterns of resistance. Additionally, sociodemographic and clinical characteristics should all be taken into account while managing patients with pneumonia empirically in this context.

The Impact of ESBLs-Positive Escherichia coli's Resistance to Cefepime and Its Guidance for Clinical Treatment.

Escherichia coli (E. coli) is a common pathogen in bloodstream infections (BSI), and the production of extended-spectrum beta-lactamases (ESBLs) is its main mechanism of resistance. However, the impact of different ESBL genotypes of E. coli on the resistance to Cefepime (FEP) remains unclear. A total of 2356 cases of BSI patients were collected. The experimental group included 188 ESBL-positive E. coli strains that were resistant to FEP but sensitive to ceftazidime (CAZ). Antibiotic usage and resistance rates were evaluated through antimicrobial susceptibility testing and antibiotic usage records. The ESBL genotypes were identified, and the minimum inhibitory concentration (MIC) and mutant prevention concentration (MPC) of FEP were determined. In ESBL-positive E. coli, three ESBL genotypes were identified: 188 strains of CTX-M, 130 strains of TEM-1, and 26 strains of OXA-10. Among them, 124 strains carried both CTX-M-9 and TEM-1 genotypes, 22 strains carried two CTX-M genotypes (CTX-M-1 and CTX-M-2), 20 strains carried both CTX-M-9 and OXA-10, and 6 strains carried three genotypes (CTX-M-9, CTX-TEM-1, and OXA-10). The MIC50, MIC90, MPC50, and MPC90 of the 188 ESBL-positive E. coli were 64, 256, 128, and 528, respectively. The MIC values ranged from 32 to 256, while the MPC values ranged from 64 to 528. The MIC50, MIC90, MPC50, and MPC90 of the 40 ESBL-negative E. coli were 0.5, 1, 64, and 128, respectively; the MIC values ranged from 0.25 to 4, while the MPC values ranged from 32 to 256, respectively. ESBL-positive E. coli induces an increase in the MIC value of FEP, leading to an increase in FEP resistance. The inoculation effect also causes a significant increase in the MPC value of FEP, especially the increase in selection index value, indicating selective enrichment and amplification of drug-resistant mutants, resulting in clinical treatment failure.