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Background and Aim: Raw milk can be a source of food-borne disease transmission and a medium for spreading antibiotic-resistant bacteria. Staphylococcus aureus and Escherichia coli are bacteria that have the pathogenic ability to attack host cells and are capable of harboring antibiotic-resistant genes. This study estimated the prevalence and antibiotic resistance of S. aureus and E. coli isolated from raw milk in East Java, Indonesia. Materials and Methods: Two hundred and fifty raw milk samples were collected from five dairy farms in East Java. S. aureus and E. coli were isolated using their respective selective media, whereas antibiotic susceptibility testing was performed using the Kirby–Bauer disk diffusion method. The methicillin-resistant S. aureus (MRSA) was confirmed using the oxacillin resistance screen agar test, and extended-spectrum beta-lactamase (ESBL)-producing E. coli was determined using the double-disk synergy test. The presence of mecA and blaTEM genes were screened by the polymerase chain reaction method. Results: Results indicated that the prevalence of S. aureus was 138 (55.2%) and that E. coli was 176 (70.4%). Of the 138 S. aureus isolated, 27 (19.6%) were MRSA, and among the 176 E. coli isolates identified, 3 (1.7%) were ESBL producers. The mecA gene was observed in 2 (7.4%) MRSA and all 3 (100%) ESBL-producing E. coli isolated harbored blaTEM genes. Conclusion: The presence of MRSA and ESBL-producing E. coli in raw milk is a serious public health threat, and public awareness should be raised about the dangers posed by these pathogenic organisms.

Introduction: In hemodialysis (HD) patients, catheter-related bloodstream infections (CRBSI) cause significant morbidity and mortality. Objectives: We analyzed dialysis CRBSI incidence, associated factors, and the spectrum of the causative organisms. Patients and Methods: Patients aged ≥18 years either on maintenance HD or with acute kidney injury having CRBSI (NKF-KDOQI criteria) were included in this prospective observational study and patients with other infections were excluded. Blood, catheter tip culture, and antibiogram were analyzed. All patients were initially treated with antibiotics covering both gram-positive and gram-negative pathogens. Results: Of 921 catheters (882 patients) included, 212 (23%) had CRBSI, of which 69(32.5%) and 143 (67.5%) had culture-positive probable CRBSI and culture-negative possible CRBSI respectively. 131 (61.8%) were

Background: The worldwide dissemination of extended spectrum beta-lactamase producing Enterobacteriales (ESBL-E) is of major concern. Microbiota may play a role in the host resistance to colonization with ESBL-E, but the underlying mechanisms remain unknown. We aimed to compare the gut microbiota composition between ESBL-producing E. coli or K. pneumoniae carriers and ESBL-E non-carriers according to the bacterial species. Results: Among 255 patients included, 11 (4,3%) were colonized with ESBL-producing E. coli and 6 (2,4%) with ESBL-producing K. pneumoniae, which were compared with age- and sex-matched ESBL-E non carriers. While no significant differences were found between ESBL-producing E. coli carriers and non-carriers, gut bacteriobiota α-diversity was decreased in ESBL-K. pneumoniae faecal carriers compared both with non-carriers (p = 0.05), and with ESBL-producing E. coli carriers. The presence of Sellimonas intestinalis was associated with the absence of ESBL-producing E. coli fecal carriage. Campylobacter ureolyticus, Campylobacter hominis, bacteria belonging to Clostridium cluster XI and Saccharomyces sp. were associated with the absence of ESBL-producing K. pneumoniae faecal carriage. Conclusions: The composition of the gut microbiota differs between ESBL-producing E. coli and K. pneumoniae faecal carriers suggesting that microbial species should be taken into account when investigating the role of gut microbiota in resistance to gut colonization with ESBL-E. Trial registration number: NCT04131569, date of registration: October 18, 2019. [ABSTRACT FROM AUTHOR]

Background: The aim of this study was to investigate the prevalence and risk factors of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae related urinary tract infections (UTI) in adult cancer patients. Methods: We conducted a retrospective study of three cancer hospitals centered on Cancer Hospital of Chinese Academy of Medical Sciences from 2015 to 2019. The clinical characters, risk factors and antimicrobial susceptibility of ESBL-producing Enterobacteriaceae UTI in adult cancer patients were described and analyzed. Results: A total of 4967 specimens of UTI were evaluated, of which 909 were positive. After excluding multiple infection bacteria, non-conforming strains, inconsistent pathological information, no drug sensitivity test or medical records, 358 episodes remained. Among them, 160 episodes belonged to ESBL-producing Enterobacteriaceae, while 198 were classified into non-ESBL group. The prevalence of ESBL UTI circled around 39.73 to 53.03% for 5 years. Subgroup analysis by tumor type revealed that 62.5% of isolates from patients with urological tumors were ESBL positive. Multivariate analysis showed that tumor metastasis (OR 3.41, 95%CI 1.84–6.30), urological cancer (OR 2.96, 95%CI 1.34–6.53), indwelling catheter (OR 2.08, 95%CI 1.22–3.55) and surgery or invasive manipulation (OR 1.98, 95%CI 1.13–3.50) were the independent risk factors. According to antimicrobial sensitivity, meropenem, imipenem and piperacillin/tazobactam were the most commonly used antibiotics for ESBL-producing Enterobacteriaceae UTI. Conclusions: In view of the high prevalence, clinicians should be alert to the occurrence of ESBL UTI, especially for patients with urological cancer or metastatic tumors. Regular replacement of urinary catheters, reduction of unnecessary invasive operations and selection of appropriate antibiotics are the necessary conditions to deal with the occurrence of ESBL UTI in adult cancer patients. [ABSTRACT FROM AUTHOR]

Background There is increasing morbidity and mortality attributed to escalating incidence of Escherichia coli bloodstream infection (BSI). The epidemiology of E. coli BSI is dynamic and differs across populations. This study aimed to describe this epidemiology in Queensland, Australia. Methods Incident E. coli BSIs (new or recurring ≥30 days from previous BSI) in adult (≥15 years) Queenslanders were identified from 2000 to 2019 using Queensland Health databases. Incidence rates, crude and standardized by age and gender, were calculated. Negative binomial regressions were performed to determine predictors of E. coli BSI incidence. Results From 2000 to 2019, 30 350 E. coli BSIs in 27 793 patients were detected; the standardized incidence rate almost doubled from 34.1 to 65.9 cases per 100 000 residents. Predictors of higher incidence rate were older age (≥65 years), comorbidity, and community-onset infection. Despite holding these factors constant, the incidence rate was estimated to increase 4% (adjusted incidence rate ratio [IRR], 1.04; 95% CI, 1.03–1.04) annually over the study period. Approximately 4.2% of E. coli isolates produced extended-spectrum beta-lactamase (ESBL-Ec), with most (95%) detected after 2010. The incidence rate of ESBL-Ec increased 25% (IRR, 1.25%; 95% CI, 1.2–1.3) annually, significantly faster than that of non-producers. Amikacin and carbapenems remain effective in vitro against ESBL-Ec BSI in Queensland. Conclusions The rise in E. coli BSIs is driven both by a higher infection rate and shifting epidemiology toward community-onset infections. These are likely attributed to an aging Australian population with increasing chronic comorbidity. The rapid expansion of ESBL-Ec in recent years is concerning and should be acknowledged for its implication in the community. [ABSTRACT FROM AUTHOR]

Background: Extended-spectrum β lactamases (ESBLs), have the ability to hydrolyze and cause resistance to various types of the β-lactam antibiotics, including the extended-spectrum (or third-generation) cephalosporins (e.g., cefotaxime, ceftriaxone, ceftazidime) and monobactams (e.g., aztreonam). ESBL-producing Gram negative bacteria is still posing significant therapeutic challenges. Objectives: To assess the prevalence and molecular characteristics of ESBL producing Gram negative bacilli, isolated from a cohort of pediatric patients in Gaza hospitals. Methods: A total of 322 isolates of Gram-negative bacilli were collected from four referral pediatric hospitals in Gaza, namely: Al-Nasr, Al-Rantisi, Al-Durra and Beit Hanoun hospitals. These isolates were tested for ESBL production using the double disk synergy and CHROMagar phenotypic methods. Molecular characterization of the ESBL producing strains was performed using PCR targeting the CTX-M, TEM and SHV genes. Antibiotic profile was done using Kirby Bauer method according to Clinical and Laboratory Standard Institute. Results: Out of 322 isolates tested by phenotypic methods, 166 (51.6%) were ESBL positive. The prevalence of ESBL production in Al-Nasr, Al-Rantisi, Al-Durra and Beit Hanoun hospitals was 54%, 52.5%, 45.5% and 52.8% respectively. The prevalence of ESBL production among Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter spp., Proteus mirabilis, Enterobacter spp., Citrobacter spp., and Serratia marcescens is 55.3%, 63.4%, 17.8%, 57.1%, 33.3%, 28.5%, 38.4%, and 4% respectively. ESBL production among urine, pus, blood, CSF and sputum was 53.3%, 55.2%, 47.4%, 33.3%, and 25% respectively. Out of the 322 isolates, 144 were screened for CTX-M, TEM and SHV production. Using PCR, 85 (59%) had at least one gene. The prevalence rate of CTX-M, TEM and SHV genes was 60%, 57.6%, and 38.3% respectively. Meropenem and amikacin were highest rates of susceptibility antibiotics against ESBLs producers (83.1% and 82.5% respectively), while the least effective antibiotics were amoxicillin (3.1%) and cephalexin (13.9%). Moreover, ESBLs producers showed high resistance rate to cefotaxime, ceftriaxone and ceftazidime (79.5%, 78.9% and 79.5% respectively). Conclusion: Our results show high prevalence of ESBL production among Gram negative bacilli isolated from children in different pediatric hospitals in Gaza strip. A substantial level of resistance to first and second generation cephalosporins was also observed. This ascertains the need for a rational antibiotic prescription and consumption policy. [ABSTRACT FROM AUTHOR]

Flomoxef is generally used to treat abdominal infections and as antibiotic prophylaxis during lower gastrointestinal surgery. It is reportedly effective against extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae and an increasingly valuable alternative to carbapenems. However, its abdominal pharmacokinetics remain unclear. Herein, pharmacokinetic analysis of flomoxef in the abdominal tissue was conducted to simulate dosing regimens for pharmacodynamic target attainment in abdominal sites. Flomoxef (1 g) was administered intravenously to a patient 30 min before commencing elective lower gastrointestinal surgery. Samples of plasma, peritoneal fluid, peritoneum, and subcutaneous adipose tissue were collected during surgery. The flomoxef tissue concentrations were measured. Accordingly, non-compartmental and compartmental pharmacokinetic parameters were calculated, and simulations were conducted to evaluate site-specific pharmacodynamic target values. Overall, 41 plasma samples, 34 peritoneal fluid samples, 38 peritoneum samples, and 41 subcutaneous adipose samples from 10 patients were collected. The mean peritoneal fluid-to-plasma ratio in the areas under the drug concentration-time curve was 0.68, the mean peritoneum-to-plasma ratio was 0.40, and the mean subcutaneous adipose tissue-to-plasma was 0.16. The simulation based on these results showed the dosing regimens (q8h [3 g/day] and q6h [4 g/day]) achieved the bactericidal effect (% T > minimum inhibitory concentration [MIC] = 40%) in all tissues at an MIC of 1 mg/L. We elucidated the pharmacokinetics of flomoxef and simulated pharmacodynamics target attainment in the abdominal tissue. This study provides evidence concerning the use of optimal dosing regimens for treating abdominal infection caused by strains like ESBL-producing bacteria. [ABSTRACT FROM AUTHOR]

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Armaghane-danesh, Yasuj University of Original Article Molecular Identification of CTX-M and SHV Betalactamase Genes in Escherichia Coli Clinical Isolates in Isfahan Moradi A, Ghiasian M*, Ghandehari F 1Department of Microbiology, Falavarjan Branch, Islamic Azad University, Isfahan, Iran Received: 25 Dec 2021 Accepted: 04 Jul 2022 Abstract Background & aim: Antimicrobial resistance is a worldwide problem causing health threats. Escherichia coli is one of the most important bacteria that causes resistance problems. The aim of the present study was to evaluate the frequency of Escherichia coli, a extended spectrum beta-lactamase producing CTX-M and SHV, in hospital and non-hospital clinical specimens in Isfahan. Methods: In the present cross-sectional descriptive study conducted in 2019, 100 samples of hospital and non-hospital of Escherichia coli strains were collected and confirmatory biochemical tests were performed to identify these strains. Resistance of strains to common antibiotics was assessed by disk diffusion method according to CLSI instructions. To perform phenotypic confirmation test of extended spectrum beta-lactamase producing strains, the combined disk method based on CLSI instructions was used. PCR was performed with specific primers to examined of encoding genes presence of SHV and CTX-M. The collected data were analyzed using statistical tests based on one-way analysis of variance and independent t-test. Results: From the studied samples, the highest antibiotic resistance was related to cefotaxime (45%) and the least antibiotic resistance was related to cefepime (39%). The results of the initial screening test showed that out of 100 Escherichia coli approved by differential tests, 38 (38%) were resistant to cephalosporin representatives and possibly produced extended-spectrum beta-lactamase. Among these samples, 33 (86.8%) after Combined disk method were confirmed as extended spectrum beta–lactamases (ESBL) producing, of which 69.7% (23 strains) had both genes encoding CTX- M and SHV had 15.2% (5 strains) of the gene encoding CTX-M and 12.1% (4 strains) had the gene encoding SHV. In one sample (3%) none of the genes encoding CTX-M and SHV were present. Conclusion: Based on the performed experiments and statistical analysis on hospital and non-hospital isolates, it was concluded that the SHV and CTX-M beta-lactamase genes in Escherichia coli had a significant increase in frequency. There was a significant relationship between SHV and CTX-M genes in hospital and non-hospital samples, so that the frequency of SHV and CTX-M genes in Escherichia coli was higher in hospital samples than non-hospital samples. There was no significant relationship between the genes identified and the type of isolates collected.

Introduction: Urinary tract infections (UTIs) are the most prevalent infections in patients worldwide. Escherichia coli is considered as the most prevalent etiological agent of UTIs. The prevalence of extended-spectrum beta-lactamase (ESBL)-producing isolates is a health care concern worldwide. The aims of this study was to determine antibiotic resistance profiles and the prevalence of the blaCTXM, blaSHV, and blaTEM genes in E. coli isolated from clinical samples obtained from patients with UTIs. Methods: From September 2019 to March 2021, a total of 1200 urine samples were collected and analyzed from patients suspected of having UTI referred to Amir-Al-Momenin hospital of Zabol, Sistan and Baluchistan province, south-east of Iran. Antibiotic resistance patterns and the prevalence of the blaCTXM, blaSHV and blaTEM genes were determined using the disk diffusion method and PCR, respectively. Results: The isolates were mostly resistance against ampicillin and trimethoprim-sulfamethoxazole, with 66% and 54% of all isolates being resistant. Also, the isolates investigated were highly susceptible against meropenem and gentamycin (95%). The prevalence of the blaCTX, blaSHV, and blaTEM genes was 22%, 30%, and 24%, respectively. Conclusion: Resistance against ampicillin and trimethoprim-sulfamethoxazole was high, therefore their prescription must be restricted. In addition, the expressions of the blaCTXM, blaSHV, and blaTEM genes were alarmingly high. In order to control the spread of infections by these isolates, constant monitoring of antibiotic resistance patterns is necessary.

Abstract Background Extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) are a well-known cause of healthcare-associated infections. The implementation of single-occupancy rooms is believed to decrease the spread of ESBL-E. Additionally, implementation of single-occupancy rooms is expected to reduce the need for intra-hospital patient transfers. We studied the impact of a new hospital with 100% single-occupancy rooms on the acquisition of ESBL-E and on intra-hospital patient transfers. Methods In 2018, the Erasmus MC University Medical Center moved from an old, 1200-bed hospital with mainly multiple-occupancy rooms, to a newly constructed 522-bed hospital with 100% single-occupancy rooms. Adult patients admitted between January 2018 and September 2019 with an expected hospitalization of ≥ 48 h were asked to participate in this study. Perianal samples were taken at admission and discharge. Patient characteristics and clinical information, including number of intra-hospital patient transfers, were collected from the patients’ electronic health records. Results Five hundred and ninety-seven patients were included, 225 in the old and 372 in the new hospital building. Fifty-one (8.5%) ESBL-E carriers were identified. Thirty-four (66.7%) patients were already positive at admission, of which 23 without recent hospitalization. Twenty patients acquired an ESBL-E, seven (3.1%) in the old and 13 (3.5%) in the new hospital building (P = 0.801). Forty-one (80.4%) carriers were only detected by the active screening performed during this study. Only 10 (19.6%) patients, six before and four during hospitalization, showed ESBL-E in a clinical sample taken on medical indication. Fifty-six (24.9%) patients were transferred to other rooms in the old hospital, compared to 53 (14.2%) in the new hospital building (P = 0.001). Intra-hospital patient transfers were associated with ESBL-E acquisition (OR 3.18, 95%CI 1.27–7.98), with increasing odds when transferred twice or more. Conclusion Transitioning to 100% single-occupancy rooms did not decrease ESBL-E acquisition, but did significantly decrease the number of intra-hospital patient transfers. The latter was associated with lower odds on ESBL-E acquisition. ESBL-E carriers remained largely unidentified through clinical samples. Trial registration This study was retrospectively registered in the Dutch National Trial Register on 24-02-2020, with registration number NL8406.

Demissew Shenkute,1 Melese Hailu Legese,2 Berhanu Yitayew,1 Asaye Mitiku,3 Getabalew Engidaye,4 Saba Gebremichael,5 Daniel Asrat,6 Yimtubezinash Woldeamanuel6 1Department of Medical Laboratory Science, College of Health Sciences, Debre Berhan University, Debre Berhan, Ethiopia; 2Department of Medical Laboratory Sciences, College of Health Sciences, Addis Ababa University, Addis Ababa, Ethiopia; 3Department of Medical Laboratory Science, College of Medicine and Health Sciences, Dilla University, Dilla, Ethiopia; 4Department of Medical Laboratory Science, Debre Berhan Health Science College, Debre Berhan, Ethiopia; 5Department of Medical Laboratory Science, College of Medicine and Health Sciences, Wollo University, Dessie, Ethiopia; 6Department of Microbiology, Immunology, and Parasitology, College of Health Sciences Addis Ababa University, Addis Ababa, EthiopiaCorrespondence: Demissew Shenkute, Email demissewshen@gmail.comBackground: Gastrointestinal colonization rate of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) is the major risk factor for infection and dissemination of resistance clones in healthcare facilities. This study aimed to investigate the magnitude of the fecal carriage of ESBL-PE and associated factors among hospitalized patients at Debre Berhan Comprehensive Specialized Hospital, North Shoa, Amhara Regional State, Ethiopia.Methods: A hospital-based cross-sectional study was conducted among 383 hospitalized patients from November 2020 to March 2021. Stool sample or rectal swab was aseptically collected and cultured on different culture media for isolation of Enterobacteriaceae. Identification was done by conventional biochemical tests. Screening of extended-spectrum beta-lactamase (ESBL) production was done by using cefotaxime and ceftazidime and confirmed by the combination disk method. Data analysis was performed by Statistical Package for Social Sciences software version 25 and a P-value ≤ 0.05 was considered as statistically significant.Results: From the total of 383 hospitalized patients, a total of 347 Enterobacteriaceae were isolated. The overall gastrointestinal colonization rate of ESBL-PE was 47.3% (164/347). The predominant ESBL-PE were E. coli 54.9% (90/164) and K. pneumoniae 33.5% (55/164). The overall multi-drug resistance rate (MDR) was 87.8% (305/347). The highest resistance was observed to ampicillin (98.3%), followed by gentamicin (80.7%), and tetracycline (73.3%), respectively. ESBL-PE were highly susceptible to meropenem (90.2%) and imipenem (89.0%). History of antibiotic use in the past 3 months (p< 0.001), admission in the neonatal intensive care unit (p=0.023), and presence of chronic disease (p< 0.001) were independently associated with fecal carriage of ESBL-PE.Conclusion: The magnitude of ESBL-PE and MDR was high in the study area. Meropenem and imipenem were active against ESBL-PE. Therefore, strict infection control measure is needed in the study area to limit the infection and dissemination of ESBL-PE.Keywords: fecal carriage, extended-spectrum beta-lactamase, Enterobacteriaceae, Ethiopia, hospitalized patients, associated factors

• Stable dominance of O25:H4/ fimH30 ST131 over a 10-y period after implementation of the '4C' antimicrobial stewardship policy. • Emergence of non-ST131 isolates within O25:H4/ fimH30. • Stable ST131 resistance to extended-spectrum beta-lactams, but significantly increased susceptibility aminoglycoside and aztreonam resistance in later years. • ST131 isolates carried a broader array of virulence genes associated with bacterial adaptation but exhibited lower and declining growth fitness over time, suggesting a trade-off between resistance and fitness. Extended-spectrum β-lactamase-producing Escherichia coli remains a major cause of hospital-acquired bloodstream infections in countries with high antimicrobial stewardship compliance. Isolates from bloodstream infections that occurred between 2010 and 2020 in a tertiary-level hospital in North-East Scotland soon after introduction of the '4C' antimicrobial stewardship policy were analysed for phylogenetic structure, antimicrobial resistance, plasmid, and virulence gene carriage. Growth fitness was measured in kinetic assays. Non-metric-multidimensional-scaling was used to evaluate clonal relationships, antimicrobial resistance, and virulence profiles in early and later years after the 4C policy introduction. Clonal and fitness trends over the study period were determined by generalised additive modelling. The relationship between clonal type, antimicrobial resistance, and fitness was evaluated by linear regression. Three hierarchical phylogenetic clusters were identified, with the most dominant cluster (O25:H4/ fimH30) including all, and nearly exclusively, Clade C ST131 isolates as well as minor non-ST131 sequence types. The prevalence of ST131 was largely stable over the study period. Resistance to aminoglycosides and aztreonam in ST131 was lower (P = 0.019 and P = 0.004, respectively) during later years (2016–2020) by 28% on average compared to early years soon after 4C policy implementation (2010–2014). Carriage of virulence factors involved in bacterial adaptation was higher in ST131 compared to non-ST131 but mostly stable in early vs. later years. Growth fitness of ST131 was lower than non-ST131 and declined steadily in later years (P < 0.0001). Despite stable virulence factor carriage, population structure, and resistance to cephalosporins, we show increased susceptibility of ST131 to aminoglycosides and aztreonam and concurrent fitness decline years after the introduction of the 4C policy. [Display omitted] [ABSTRACT FROM AUTHOR]

The increasing prevalence of antimicrobial resistance and the limited availability of new antimicrobial agents have created an urgent need for new approaches to combat these issues. One such approach involves reevaluating the use of old antibiotics to ensure their appropriate usage and maximize their effectiveness, as older antibiotics could help alleviate the burden on newer agents. An example of such an antibiotic is chloramphenicol (CHL), which is rarely used due to its hematological toxicity. In the current study, we employed a previously published transposon mutant library in MG1655/pTF2::blaCTX-M-1, containing over 315,000 unique transposon insertions, to identify the genetic factors that play an important role during growth in the presence of CHL. The list of conditionally essential genes, collectively referred to as the secondary resistome (SR), included 67 genes. To validate our findings, we conducted gene knockout experiments on six genes: arcA, hfq, acrZ, cls, mdfA, and nlpI. Deleting these genes resulted in increased susceptibility to CHL as demonstrated by MIC estimations and growth experiments, suggesting that targeting the products encoded from these genes may reduce the dose of CHL needed for treatment and hence reduce the toxicity associated with CHL treatment. Thus, the gene products are indicated as targets for antibiotic adjuvants to favor the use of CHL in modern medicine.

Mihret Tilahun, Alemu Gedefie, Habitye Bisetegn, Habtu Debash Department of Medical Laboratory Sciences, College of Medicine and Health Science, Wollo University, Dessie, EthiopiaCorrespondence: Mihret Tilahun, Department of Medical Laboratory Science College of Medicine and Health Sciences, Wollo University, PO.Box: 1145, Dessie, Ethiopia, Tel +251 920988307, Fax +251 333115250, Email tilahunmihret21@gmail.comBackground: The emergence of extended spectrum beta lactamase and carbapenemase production of Acinetobacter and Pseudomonas aeruginosa is a great concern and major cause of nosocomial infections due to its ability to production of extended spectrum beta lactamase and carbapenemase enzymes.Objective: To assess Emergence of high prevalence of extended-spectrum beta-lactamase and Carbapenemase producing Acinetobacter species and Pseudomonas aeruginosa among hospitalized patients at Dessie Comprehensive Specialized Hospital, North-East Ethiopia.Materials and Methods: A hospital-based cross-sectional study was conducted from February–August 2021 at Dessie Referral Hospital in the North eastern Ethiopia. A total of 423 clinical samples taken from admitted patients. Clinical specimens were collected aseptically and inoculated on blood agar and MacConkey agar media. Antimicrobial susceptibility test, ESBL and carbapenemase production were performed as CLSI guideline. The data were entered into the Epi-data and imported to Statistical Package for Social Science version 25. P value of < 0.05 with odds ratio and 95% confidence interval was considered as statistically significant.Results: Out of 423 clinical specimens 17.7% (75/423) were culture positive nosocomial infection. The proportions of nosocomial infection were higher in blood stream followed by wound infection which accounted 24.6%, 20.8%, respectively. Overall, 32/75 (42.7%) and 19/75 (25.3%) patients had infection with ESBL and CP producing bacterial infection. P. aeruginosa was the most predominant isolated bacteria 46/75 (52.9%). The overall multidrug resistance rate of the isolated bacteria was 88% (66/75). The majority of highest resistance rate was Piperacillin tazobactam 50 (66.7%) and Aztreonam 26 (56.5%), respectively, while least resistance rate was Amikacin 27 (36%).Conclusion: The incidence rates of ESBL, carbapenemase production and antimicrobial resistant Acinetobacter species and P. aeruginosa infections are high. Therefore, treatment should be based on culture and antimicrobial test result and minimize the use of antibiotics empirically.Keywords: nosocomial infections, extended-spectrum beta-lactamase, carbapenemase, Acinetobacter, Pseudomonas aeruginosa, drug resistance

ABSTRACT: Objectives: This study aimed to clarify the characteristics of Tn7-derivatives transposons in MDR Proteus mirabilis strains isolated from anal swabs of chicken and swine in China from 2015–2020. Methods: The Tn7 tnsA gene was screened in 207 P. mirabilis isolates by polymerase chain reaction (PCR). These strains were subjected to antimicrobial susceptibility testing. Illumina Hiseq (200 × coverage) was used for genome sequencing. Transposon maps were completed by PCR and Sanger sequencing and analysed by BLAST. Results: The Tn7 tnsA gene was detected in 21 strains by PCR. Eight novel Tn7-derivatives, named Tn6667, Tn6668, Tn6669, Tn6670, Tn7095, Tn7096, Tn7097 and Tn7098, were characterised. Three types of hybrid class 2/1 integrons were found at the right end of Tn7 derivatives. A novel Tn7-like transposon Tn6666 with an active integrase gene intI2, whose transposition module shows 93% nucleotide identity to the corresponding region of Tn7, was characterised in three strains. Tn6666 is also found next to Tn7097 or Tn7098 in the chromosomes of two clonally related P. mirabilis strains. The number of resistance genes carried by the novel transposons varied from 1 to 18. A novel variant of class A extended-spectrum beta-lactamase gene, blaPER-16, with eight base substitutions compared with blaPER-12, was harboured by Tn7098. Conclusion: Our study characterised diverse novel Tn7-derivatives and a new Tn7-like transposon in P. mirabilis. An active integrase gene intI2 might promote the diversification of Tn7-like transposons. More attention should be paid to the prevalence and evolution of Tn7-derivatives and Tn7-like transposons and antimicrobial resistance genes they carry.

Abstract Background Urinary tract infections (UTIs), which are usually caused by bacteria in the Enterobacterales family, are a common reason for outpatient visits. Appropriate empiric therapy for UTIs requires an understanding of antibiotic resistance in the community. In this nationwide study, we examined trends in antibiotic resistance in urinary Enterobacterales isolates from ambulatory patients in the United States (US). Methods We analyzed the antimicrobial susceptibility profiles (extended-spectrum beta-lactamase [ESBL]-producing phenotype and not susceptible [NS] to beta-lactams, trimethoprim/sulfamethoxazole [TMP/SMX], fluoroquinolones [FQ], or nitrofurantoin [NFT]) of 30-day non-duplicate Enterobacterales isolates from urine cultures tested at ambulatory centers in the BD Insights Research Database (2011–2020). The outcome of interest was the percentage of resistant isolates by pathogen and year. Multi-variable generalized estimating equation models were used to assess trends in resistance over time and by additional covariates. Results A total of 338 US facilities provided data for > 2.2 million urinary Enterobacterales isolates during the 10-year study. Almost three-quarters (72.8%) of Enterobacterales isolates were Escherichia coli. Overall unadjusted resistance rates in Enterobacterales isolates were 57.5%, 23.1%, 20.6%, and 20.2% for beta-lactams, TMP/SMX, FQ, and NFT, respectively, and 6.9% had an ESBL-producing phenotype. Resistance to two or more antibiotic classes occurred in 16.4% of isolates and 5.5% were resistant to three or more classes. Among isolates with an ESBL-producing phenotype, 70.1%, 59.9%, and 33.5% were NS to FQ, TMP/SMX, and NFT, respectively. In multivariable models, ESBL-producing and NFT NS Enterobacterales isolates increased significantly (both P

Background: Lower respiratory tract infections (LRTIs) are one of the most frequent infections seen in humans. Bacterial etiology of these infections is one of the major causes of morbidity and mortality. The emergence of drug resistance among bacteria is increasing throughout the world causing more severe infections because of their continuous mutation and multidrug resistance nature. Objectives: This study was focused on obtaining a comprehensive insight into the Gram-negative bacillary profile of LRTIs, their prevalence, and their antibiotic susceptibility patterns. Materials and Methods: The study was conducted for a duration of 6 months. Samples were obtained from patients of all ages and both sexes presenting with symptomatology suggestive of LRTIs. Following conventional culture, the isolated organisms were identified by various preliminary identification methods and biochemical tests. Antimicrobial sensitivity testing of Gram-negative isolates was performed by standard methods as recommended by CLSI 2019. Results: Out of the 1724 LRT specimens evaluated, 307 (17.80%) were culture positive. Our study showed that Gram-negative bacilli are the predominant cause (97.70%) of LRTIs with Klebsiella pneumoniae (42%) as the major pathogen followed by Escherichia coli (31.66%), Pseudomonas aeruginosa (25.33%), and Acinetobacter baumannii (1%). Extended-spectrum beta-lactamase production was detected in 3.33% and metallo-beta-lactamase in 2% of the total GNB pathogens. Conclusion: For effective management of LRTIs, an ultimate and detailed bacteriological diagnosis along with antimicrobial susceptibility testing is required to overcome the global problem of antibiotic resistance. [ABSTRACT FROM AUTHOR]

Transplant recipients are highly susceptible to multidrug-resistant (MDR) related infections. The lack of early appropriate antimicrobial treatment may contribute to the high mortality due to MDR-related infections in transplant recipients especially in case of metallo-β-lactamases. In this review, we present the current state of knowledge concerning multidrug-resistant Gram negative bacilli's risk management in the care of solid-organ transplant recipients and suggest control strategies. We searched for studies treating MDR g-negative bacilli related infections in the renal and hepatic transplant patient population. We included randomized and observational studies. Solid-organ transplant is the best therapeutic option for patients diagnosed with end-stage organ disease. While the incidence of opportunistic infections is decreasing due to better prevention, the burden of "classical" infections related to MDR bacteria especially related to Gram-negative bacteria is constantly increasing. Over the last two decades, various MDR pathogens have emerged as a relevant cause of infection in this specific population associated with significant mortality. Several factors related to the management of transplant donor candidates and recipients increase the risk of MDR infections in transplant recipients. The awareness of this high susceptibility of transplant recipients to MDR-related infections challenges the choice of empirical therapy, while its appropriateness can only be validated a posteriori. Indeed, the lack of early appropriate antimicrobial treatment may contribute to the high mortality due to MDR-related infections in transplant recipients especially in case of metallo-β-lactamases. Multidrug-resistant Gram-negative bacteria are associated with high morbidity and mortality in solid organ transplant recipients. It seems important to identify patients at risk of colonization/MDR bacteria to evaluate strategies to limit the risk of secondary infections and to minimize the inappropriate use of broad-spectrum antibiotics. [ABSTRACT FROM AUTHOR]

Background: Avibactam, relebactam and vaborbactam are β-lactamase inhibitors that proved their efficiency against KPC-producing Enterobacterales. Regarding their inhibitor activity towards Ambler's class A extended spectrum β-lactamases (ESBL) and Ambler's class C cephalosporinase (AmpC), they should be active on most of the carbapenem-resistant non-carbapenemase-producing Enterobacterales (CR non-CPE). Objectives: Determine the in vitro activity of ceftazidime-avibactam, imipenem-relebactam and meropenem-vaborbactam and comparators against CR non-CPE. Methods: MICs to ceftazidime/avibactam, imipenem/relebactam, meropenem/vaborbactam, but also temocillin, ceftolozane/tazobactam, ertapenem, colistin, eravacycline and tigecycline were determined by broth microdilution (ThermoFisher) on a collection of 284 CR non-CPE (inhibition zone diameter < 22 mm to meropenem). Whole genome sequencing was performed on 90 isolates to assess the genetic diversity as well as resistome. Results: According to EUCAST breakpoints, susceptibility rates of ceftazidime, imipenem, meropenem and ertapenem used at standard dose were 0.7%, 45.1%, 14.8% and 2.5%, respectively. Increased exposure of ceftazidime, imipenem and meropenem led to reach 3.5%, 68.3% and 67.7% susceptibility, respectively. Using the EUCAST clinical breakpoints, susceptibility rates of ceftazidime/avibactam, imipenem/relebactam and meropenem/vaborbactam were 88.4%, 81.0% and 80.6%, respectively. Susceptibility rates of temocillin, ceftolozane/tazobactam, tigecycline, eravacycline, and colistin were 0%, 4.6%, 27.8%, 54.9% and 90.1%. MICs distributions with and without the presence of the inhibitor demonstrated a better ability of avibactam and relebactam compared to vaborbactam to restore susceptibility to the associated β-lactam. Conclusions: This study demonstrated the in vitro efficacy of ceftazidime/avibactam, imipenem/relebactam and to a lesser extent meropenem/vaborbactam against CR non-CPE. Moreover, to test all β-lactams/β-lactamases inhibitors combinations without a priori for CRE, non-CPE is crucial since resistance to one of the β-lactam/β-lactamase inhibitor combinations does not predict resistance to another molecule, depending on the resistance mechanisms involved. [ABSTRACT FROM AUTHOR]

The spread of extended-spectrum beta-lactamase (ESBL)-producing bacteria in the environment has been recognized as a challenge to public health. The aim of the present study was to assess the occurrence of ESBL-producing Escherichia coli and Klebsiella pneumoniae from selected water bodies receiving hospital effluents in Kerala, India. Nearly 69.8% of Enterobacteriaceae isolates were multi-drug resistant by the Kirby-Bauer disc diffusion method. The double disc synergy test was used to detect the ESBL production and the genes responsible for imparting resistance were detected by PCR. Conjugation experiments confirmed the mechanism of plasmid-mediated transfer of resistance. The prevalence of ESBL production in E. coli and K. pneumoniae was 49.2 and 46.8%, respectively. Among the ESBL-encoding genes, blaCTX-M was the most prevalent group followed by blaTEM, blaOXA, blaCMY, and blaSHV. The results suggest that healthcare settings are one of the key contributors to the spread of ESBL-producing bacteria, not only through cross-transmission and ingestion of antibiotics but also through the discharge of waste without a proper treatment, leading to harmful effects on the aquatic environment. The high prevalence of ESBL-producing Enterobacteriaceae with resistance genes in public water bodies even post-treatment poses a serious threat. [ABSTRACT FROM AUTHOR]

Background: Extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-E) are a well-known cause of healthcare-associated infections. The implementation of single-occupancy rooms is believed to decrease the spread of ESBL-E. Additionally, implementation of single-occupancy rooms is expected to reduce the need for intra-hospital patient transfers. We studied the impact of a new hospital with 100% single-occupancy rooms on the acquisition of ESBL-E and on intra-hospital patient transfers. Methods: In 2018, the Erasmus MC University Medical Center moved from an old, 1200-bed hospital with mainly multiple-occupancy rooms, to a newly constructed 522-bed hospital with 100% single-occupancy rooms. Adult patients admitted between January 2018 and September 2019 with an expected hospitalization of ≥ 48 h were asked to participate in this study. Perianal samples were taken at admission and discharge. Patient characteristics and clinical information, including number of intra-hospital patient transfers, were collected from the patients' electronic health records. Results: Five hundred and ninety-seven patients were included, 225 in the old and 372 in the new hospital building. Fifty-one (8.5%) ESBL-E carriers were identified. Thirty-four (66.7%) patients were already positive at admission, of which 23 without recent hospitalization. Twenty patients acquired an ESBL-E, seven (3.1%) in the old and 13 (3.5%) in the new hospital building (P = 0.801). Forty-one (80.4%) carriers were only detected by the active screening performed during this study. Only 10 (19.6%) patients, six before and four during hospitalization, showed ESBL-E in a clinical sample taken on medical indication. Fifty-six (24.9%) patients were transferred to other rooms in the old hospital, compared to 53 (14.2%) in the new hospital building (P = 0.001). Intra-hospital patient transfers were associated with ESBL-E acquisition (OR 3.18, 95%CI 1.27–7.98), with increasing odds when transferred twice or more. Conclusion: Transitioning to 100% single-occupancy rooms did not decrease ESBL-E acquisition, but did significantly decrease the number of intra-hospital patient transfers. The latter was associated with lower odds on ESBL-E acquisition. ESBL-E carriers remained largely unidentified through clinical samples. Trial registration: This study was retrospectively registered in the Dutch National Trial Register on 24-02-2020, with registration number NL8406. [ABSTRACT FROM AUTHOR]

Background: Efforts to combat antimicrobial resistance (AMR) should be based on the One Health approach, involving human health, animal health, and the environment. In Ghana, previous studies on AMR have given little attention to animal source food, a major route of transmission of antibiotic-resistant zoonotic pathogens. The aim of this study was to investigate the occurrence of multidrug-resistant (MDR) bacteria in meat sold in Accra. Methods: This was a cross-sectional study in which 270 meat samples (90 each of beef, goat meat, and chicken) were collected, and investigated for contamination with multidrug-resistant bacteria. The bacteria were subjected to susceptibility testing against amikacin (30 µg), ampicillin (10 µg), amoxicillin-clavulanate (20/10 µg), cefuroxime (30 µg), ceftriaxone (30 µg), ceftazidime (30 µg), cefepime (30 µg), ciprofloxacin (5 µg), trimethoprim-sulfamethoxazole (1.25/23.75 µg), ertapenem (10 µg), meropenem (10 µg), imipenem (10 µg), tigecycline (15 µg), and gentamicin (10 µg). Results: Thirty-two different types of bacteria, totalling 558, were isolated, the predominant being Escherichia coli (44.6%), Aeromonas hydrophila (19.9%), Vibrio cholerae (3.4%), Aeromonas veronii (3.2%), and Klebsiella pneumoniae (3.1%). The prevalence of MDR among the contaminating bacteria was 14.9%. The MDR distribution among the predominant bacteria was Escherichia coli (18.7%), Aeromonas hydrophila (11.1%), Vibrio cholerae and Aeromonas veronii (0.0% each), and K. pneumoniae (5.6%). Moreover, 2.0% of the contaminating bacteria were extended-spectrum beta-lactamase (ESBL) producers, all of which occurred in the chicken samples, and their distribution was: Escherichia coli (1.3%), Klebsiella pneumoniae, Pantoea spp., Enterobacter cloacae, and Serratia plymuthica (0.2% each). Conclusions: The meat samples were heavily contaminated with Escherichia coli and Aeromonas hydrophila, and less frequently, with Vibrio cholerae, Klebsiella pneumoniae, and other organisms. The prevalence of multidrug-resistant bacteria was moderate (14.9%), while that of ESBL producers was low (2%). [ABSTRACT FROM AUTHOR]

The aim of our study was to delineate an outbreak of gastroenteritis caused by Shigella flexneri and affecting sixteen persons between May and June 2014 in Bilbao, Spain. All patients exhibited symptoms after consuming kebab in the same kebab shop. The outbreak is described through the clinical cases, the microbiological and molecular genetic diagnosis, and the epidemiologic investigation. Minimum inhibitory concentrations for ampicillin, amoxicillin plus clavulanic acid, third and fourth generation cephalosporins, carbapenems, monobactams, aminoglycosides, fluoroquinolones, co-trimoxazole, colistin and tigecycline were measured. The S. flexneri strains were screened by PCR for TEM, SHV, CTX-M beta-lactamases and plasmidic AmpCs and aac(6′)-Ib gene. Serotyping, pulsed field gel-electrophoresis, conjugation assay, plasmid sizing by S1 enzyme digestion and Southern blot hybridization were accomplished. All the S. flexneri isolates proved to be serotype 2 and produced extended-spectrum beta-lactamase (ESBL). Carbapenems, fluoroquinolones, tigecycline, colistin, and co-trimoxazole remained active antibiotics. All the strains harboured bla CTX-M-15 and bla OXA-1 genes. The strains hosted two high-molecular weight plasmids of 100 and 230 kb, respectively. According to the hybridization assay bla CTX-M-15 was located on the plasmid of 230 kb. The identical pulsotype verified the presence of outbreak. Remarkable, that one of the food handlers has travelled recently to Pakistan, where ESBL-producing Shigella strains had been reported previously. To the best of our knowledge, this is the first outbreak caused by CTX-M-15-expressing S. flexneri in Spain and as well as in Europe. [ABSTRACT FROM AUTHOR]

BACKGROUND: This study aimed to determine the prevalence of extended-spectrum-beta-lactamase (ESBL) and carbapenem-resistant gram-negative bacteria (GNB) isolated from patients at the King Faisal Hospital in Makkah, Saudi Arabia. METHODS: In this cross-sectional study, a total of 298 patients admitted to the intensive care unit for 48 hours and who had a central venous catheter were selected using a census sampling method. Only patients with ESBL and carbapenem-resistant GNB-isolated organisms (175 patients) were included. The susceptibility test of GNB was carried out according to the standard recommendations. The identified strains were tested in-vitro against several antimicrobial drugs. Statistical analysis was performed using SPSS version 24. RESULTS: 36(20.6%) of samples were ESBL-producing GNB, whereas 139(79.4%) were carbapenem-resistant GNB. The pooled proportional estimates of ESBL-producing GNB Escherichia coli, Klebsiella pneumoniae, and other GNB were 44.4%, 41.6%, and 14.0%, respectively; the pooled proportional estimates of carbapenem resistance GNB Klebsiella pneumoniae, Acinetobacter baumannii complex/hemolyticus and other GNB were 82.8%, 10.8%, and 6.4%, respectively. All ESBL-producing GNB and carbapenem-resistance GNB were multidrug-resistant pathogens. The highest carbapenem resistance GNB 139(100%) was to ampicillin, and the lowest 122(87.7%) was to Amoxicillin/clavulanic acid (Amox/clav). All ESBL-producing GNB 36 (100%) were resistant to cefotaxime, and 35 (97.2%) were resistant to ampicillin, cefuroxime, cefepime, and ceftazidime. Additionally, the effective antibiotic against ESBL-producing GNB was imipenem. CONCLUSION: Antibiotic utilization measures appear to contribute to the control of the emergence of multidrug-resistant pathogens such as ESBL and carbapenem-resistant GNB. Strict adherence to well-accepted infection control guidelines along with caution in using broad-spectrum antimicrobial agents represents the best strategy for preventing the emergence and spread of multidrug-resistant pathogens. [ABSTRACT FROM AUTHOR]

Background and Aim: Acinetobacter baumannii is one of the most important pathogens found in ICUs as it is responsible for nosocomial infection with a high mortality rate and a rising level of resistance to most antibiotics. One of the main mechanisms of resistance is the production of extended-spectrum beta-lactamase (ESBL) genes such as veb, per, tem, and shv. The current study aimed to determine the antibiotic resistance pattern and the frequency of per, tem, veb, and shv genes in A. baumannii strains isolated from Zahedan, Iran. Materials and Methods: The current study was conducted on 150 strains of A. baumannii isolated from different clinical samples from January to September 2018. The antibiotic resistance pattern was determined by the disk diffusion method for 18 antibiotics and the minimum inhibitory concentration for colistin. Later, the bacterial genome was extracted, and PCR detected ESBL genes. Results: Out of the 150 studied isolates, 141 were A. baumannii and only nine isolates were A. nosocomialis. A. baumannii strains were strongly resistant to many selected antibiotics such as CAZ (99.3%), CTX (97.9), PTZ (97.2%), SXT (97.2%), IMI (97.2%), and LEV (96.5%); while this value was low for a few antibiotics including MN (69.5%), DXT (52.5%), SAM (21.3), and TN (16.7). The eb, per, tem, and shv genes were detected in 20 (13.3%), 15 (10%), 23 (15.4%), and 11 (7.3%) isolates, respectively. Conclusion: The isolates were highly resistant to ceftazidime, ceftriaxone, piperacillin-tazobactam, cotrimoxazole, imipenem, and levofloxacin; in addition, the frequency of per, veb, tem, and genes was lower in the current study than those reported in other regions. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas aeruginosa is a human opportunistic pathogen that is known to be responsible for various diseases. However, its antibiotic-resistant isolates often cause serious infections. Methods: This study for the first time investigated a total of 80 P. aeruginosa isolates collected from patients admitted to Amir AlMomenin hospital. The isolates were identified by biochemical assays. The combination disc test method was used to measure antibiotic susceptibility and confirm the presence of extended spectrum-beta lactamases-producing enzymes. Also, the presence of enzymeproducing genes bla CTXM-1, bla CTXM-2, bla CTXM-3, bla SHV, and bla OXA of the target enzymes was examined using polymerase chain reaction. Results: Out of 80 P. aeruginosa isolates, 32 isolates (40%) were beta-lactamase generators. Resistance to the studied antibiotics was found to be 97.5%, 90%, 81.3%, 75%, 75%, 72.5%, 60%, 52.5%, 50%, 32.5%, 28.8%, and 0% for amoxicillin, amoxiclav cephalexin, nitrofurantoin, cotrimoxazole, azithromycin, ceftriaxone, cefotaxime, gentamicin, ceftazidime, ciprofloxacin, and imipenem, respectively. Therefore, the highest antibiotic resistance was against amoxicillin, co-amoxiclav, and cephalexin, respectively, while the lowest was detected for imipenem. Besides, 17.5% of the studied isolates were multidrug-resistant (MDR). Among extended-spectrum beta-lactamases-producing genes, bla CTXM-3 displayed the highest frequency of 84.4%. Conclusion: The findings demonstrated the wide resistance of P. aeruginosa isolates against various antibiotic classes. According to the results, it is suggested to identify different patterns of antibiotic resistance of P. aeruginosa isolates prior to the onset of treatment for any P. aeruginosa-related infections. [ABSTRACT FROM AUTHOR]

Background: Bacteremia caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli ( E. coli) can lead to bloodstream infection and subsequent sepsis which increases morbidity and mortality. Evaluation of risk factors of infection by ESBL-producing E. coli is important as it can decrease inappropriate antibiotic use and mortality rates. This study aimed to identify the risk factors and mortality of bacteremia caused by ESBL-producing E. coli. Methods: This retrospective cohort study included inpatients with confirmed E. coli blood culture examinations from two referral hospitals in Jakarta, Indonesia. Data suspected as risk factors for ESBL-producing E. coli bacteremia (utilization of medical devices, age, Charlson Comorbidity Index, history of hospitalization, and history of antibiotic therapy) were collected for analysis. Clinical profiles and independent risk factors of ESBL-producing E. coli bacteremia associated mortality were also evaluated. Results: A total of 116 subjects were analyzed with 81% aged ≥18 years old. The most common source of infection was the gastrointestinal and intra-abdominal tracts. Malignancy as comorbidity was present in 46.6% subjects. Significant risk factors for developing ESBL-producing E. coli bacteremia were history of antibiotic therapy and utilization of medical devices. The proportion of mortality in ESBL-producing E. coli bacteremia was 55.7% with age and sepsis as its independent risk factors. Conclusions: History of antibiotic therapy and utilization of medical devices were significant risk factors for ESBL-producing E. coli bacteremia. The proportion of mortality in ESBL-producing E. coli bacteremia patients was 55.7% with its independent risk factors being age and sepsis.

Background: Escherichia coli (E. coli)  infections are becoming difficult to treat due to the bacterium's biofilm-forming capabilities and rising resistance to multiple antibiotics, posing a growing clinical challenge. This study assessed the antimicrobial resistance and biofilm formation by Escherichia coli isolates from patients at a hospital in Tigray, Northern Ethiopia., Method: From patients exhibiting signs of bacterial infection, while excluding recent antibiotic users or those with incomplete data, 417 clinical samples comprised of 84 blood, 108 pus, and 225 urine samples were obtained in a cross-sectional study. The combination disc method was used to test extended-spectrum beta-lactamase (ESBL) production, and Ampicillin C (AmpC) enzyme presence was confirmed with cefoxitin and cefotaxime discs. Data analysis was conducted with SPSS version 22 software, applying ANOVA and logistic regression, with significance set at p<0.05., Result: Among the 417 samples, 109 (26.1%) tested positive for Escherichia coli . These isolates showed high resistance to ampicillin (84.4%) but lower resistance to meropenem (9.17%). ESBL was detected in 46.8% of isolates and AmpC in 54%, with 48 (44%) isolates positive for both. Strong biofilm formation occurred in 76% of isolates, while only 2.75% were weak producers. Biofilm strength correlated significantly with prior antibiotic use (p=0.028), ward type (p=0.001), and use of indwelling devices (p=0.000)., Conclusion: In northern Ethiopia, Escherichia coli isolates showed resistance to major antibiotic classes like beta-lactams, fluoroquinolones, and aminoglycosides. This high resistance and biofilm development highlight the critical need for interventions to curb resistance spread, with a focus on antibiofilm research and enhanced infection prevention measures., Competing Interests: Human subjects: Consent for treatment and open access publication was obtained or waived by all participants in this study. Institutional Review Board of the College of Health Sciences, Addis Ababa University issued approval 033/19/SoP. Animal subjects: All authors have confirmed that this study did not involve animal subjects or tissue. Conflicts of interest: In compliance with the ICMJE uniform disclosure form, all authors declare the following: Payment/services info: All authors have declared that no financial support was received from any organization for the submitted work. Financial relationships: All authors have declared that they have no financial relationships at present or within the previous three years with any organizations that might have an interest in the submitted work. Other relationships: All authors have declared that there are no other relationships or activities that could appear to have influenced the submitted work., (Copyright © 2024, Gebremariam et al.)

Background: The increasing prevalence of multidrug-resistant (MDR) Escherichia coli strains, especially extended-spectrum beta-lactamase (ESBL)-producing strains, has become a global health concern. This study was aimed to determine the frequency of blaCTX-M, blaTEM, and blaSHV genes among E. coli isolates from urinary tract infection (UTI) and evaluate their antibiotic resistance pattern. Methods: Totally 98 E. coli isolates were recovered from urine samples of UTI-diagnosed patients. Antibiotic resistance and ESBL production were evaluated by disk diffusion and combined disk methods according to the Clinical Laboratory Standards Institute guidelines. The biofilm formation ability of isolates was assessed using the tube adherence method. ESBL-positive isolates were screened for blaTEM, blaCTX-M, and blaSHV genes by polymerase chain reaction. Results: Among the examined isolates, 25 (25.5%) were detected as ESBL producers and harbored at least one of the studied genes. The blaCTX-M was the predominant (44%) gene, followed by blaTEM (24%) and blaSHV (8%). The isolates revealed variable resistance levels to all antimicrobials, out of which 55.1% were conferred a high resistance rate to different antibiotic classes and considered MDR. Phenotypically, 42.85% of the isolates were biofilm formers, of which the majority (38%) formed moderate biofilms. Conclusions: This study showed that the ESBL-positive isolates were more resistant to some first-line antibiotics, and this highlights the necessity to control and monitor the prescribed antibiotics used for empirical treatment for UTI patients.

Aim: In this prospective study we aimed to compare the effectiveness of VITEK-2 (bioMérieux, France) automated system, double-disc synergy (DDST) versus combined disc test (CDT) in detecting the Extended-Spectrum Beta-Lactamase (ESBL) positivity in Escherichia coli and Klebsiella pneumoniae strains isolated from various clinical samples. Material and Method: E.coli and K. pneumoniae strains inoculated on Mueller Hinton Agar plate. Susceptibility tests were performed with the VITEK 2 (BioMérieux, France) system before. Afterward, EBSL positivity was investigated manually DDST and CDT. Minimal inhibitor concentration (MIC) results of three tests were compared with each other according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) criteria. Results: 184 E. coli and K.pneumoniae strains, elevated. 92.9% of 98 patients with VITEK 2 positive results were positive with combined disc and DDS method, 100% of the 86 patients with negative results of VITEK 2 were negative with combined disc and MDC. Conclusion: VITEK 2 was found to have a sensitivity of 100%, a specificity of 92.4%, a positive predictive value of 92.8% and a negative predictive value of 100%. VITEK 2 was found to be compatible with validation tests for ESBL positivity.

Background and Objectives: Upper urinary tract infections (UTIs) that require in-patient care can be expensive. Comorbid conditions such as diabetes as well as UTI due to extended-spectrum beta-lactamase (ESBL) producing bacteria may affect costs. The quality of life of patients with this condition has not been described. Methods: This was a cost of illness study that prospectively evaluated patients admitted with upper UTI to a medical ward in a tertiary care hospital. Direct medical and nonmedical costs, indirect costs were collected to make the total cost per admission. Quality of life was assessed using the World Health Organization Quality of Life-BREF score. We also compared costs between those with and without diabetes or ESBL infection. Results: Between March 2016 and July 2017, 92 eligible patients were included in the study. The average age was 55.8 years; two thirds were diabetics. The mean overall cost of a single admission for upper UTI was INR.88, 330.2 (1370.4 USD). This was INR.96, 193.0 (1492.6 USD) and INR.1, 03,154.9 (1600.4 USD) among those with diabetes mellitus and ESBL infection, respectively. The cost was higher among those with diabetes and ESBL than those without; this difference reached statistically significance for the ESBL group. The quality of life was affected in all domains; the psychological being most affected among diabetics and ESBL infected. Conclusions: The mean total cost of admission for an upper UTI in a tertiary care hospital in South India was INR 88,330 (1370.4 USD). This is higher if the patient has diabetes or ESBL organism causing the UTI. Quality of life is clearly reduced especially in the psychological domain.

Background and Aim: The production of beta-lactamase enzymes, such as extended-spectrum beta-lactamase (ESBL), adenosine-monophosphate-cyclic (AmpC), and Klebsiella pneumoniae carbapenemase (KPC), is one of the most important mechanisms of bacterial resistance to antimicrobials. Gram-negative bacteria show significant resistance due to various intrinsic and acquired factors. These intrinsic factors include low permeability of the outer membrane, various efflux systems, and the production of beta-lactamases, while acquired factors include chromosomal mutation and acquisition of resistance genes by horizontal transfer. Mobile elements such as plasmids, integrative conjugative elements, mobilizable islands, or transposable elements are involved in horizontal transfer. At present, the Gram-negative pathogens of most concern are Acinetobacter baumannii, Pseudomonas aeruginosa, and those belonging to the Enterobacteriaceae family (e.g., Escherichia coli, K. pneumoniae, and Proteus mirabilis). This study aimed to evaluate the profile of antimicrobial resistance and the production of the enzymes ESBL, AmpC, and KPC, in 21 gram-negative bacteria isolated from domestic animals treated at the University Veterinary Hospital (HVU) of the Federal University of Western Bahia (UFOB). Materials and Methods: The biological samples (21) were inoculated to brain heart infusion broth, blood agar, and MacConkey agar and incubated for 24-72 h at 37°C. Gram staining and identification through biochemical tests and matrix-associated laser desorption/ionization time-of-flight mass spectrometry were conducted. To evaluate the antimicrobial resistance profile, the disk diffusion method was used, and 25 antibiotics were employed. For the detection of ESBL, the disk approximation method was applied using chromogenic agar. The presence of KPC was observed using chromogenic agar and the Hodge test. For AmpC evaluation, the disk approximation method was used. Results: The most isolated agent was E. coli (66.66%, 14/21), followed by K. pneumoniae and P. mirabilis (both 14.29%, 3/21), and then Pasteurella spp. (4.76%, 1/21). The bacterial isolates showed high levels of resistance against clindamycin, penicillin, imipenem, polymyxin, cefoxitin, gentamycin, cefotaxime, ceftazidime, cephalothin, ceftriaxone, ciprofloxacin, trimethoprim/sulfamethoxazole, chloramphenicol, and tetracycline. The best effectiveness rates were observed for cefepime, streptomycin, amoxicillin-clavulanate, aztreonam, nalidixic acid, tobramycin, levofloxacin, amikacin, and meropenem. All biological isolates showed multiple resistance to at least three of the antibiotics tested (3/25), and some showed resistance to 24 of the antibiotics tested (24/25). Among the 21 pathogens analyzed, 8 were ESBL producers (38.09%); of these, 6 were identified as E. coli (28.57%), and 2 were identified as K. pneumoniae (9.52%). Two strains of K. pneumoniae produced both ESBL and KPC. None of the isolates were producers of AmpC. Conclusion: The results found in the present work raise concern about the level of antimicrobial resistance among pathogens isolated from domestic animals in Brazil. The results highlight the need for the development and implementation of anti-resistance strategies to avoid the dissemination of multiresistant pathogens, including the prudent use of antimicrobials and the implementation of bacterial culture, antimicrobial sensitivity, and phenotypic tests for the detection of beta-lactamase enzymes in bacteria isolated from animals.

Abstract Background Increasing rates of antibiotic resistance in Gram-negative organisms due to the presence of extended-spectrum beta-lactamases (ESBL), hyperproduction of AmpC enzymes, carbapenemases and other mechanisms of resistance are identified in common hospital- and healthcare-associated pathogens including Enterobacteriaceae, Pseudomonas aeruginosa and Acinetobacter baumannii. Cefiderocol is a novel siderophore cephalosporin antibiotic with a catechol moiety on the 3-position side chain. Cefiderocol has been shown to be potent in vitro against a broad range of Gram-negative organisms, including carbapenem-resistant Enterobacteriaceae (CRE) and multi-drug-resistant (MDR) P. aeruginosa and A. baumannii. Recent clinical data has shown cefiderocol to be effective in the setting of complicated urinary tract infections and nosocomial pneumonia, but it has not yet been studied as treatment of bloodstream infection. Methods This study will use a multicentre, open-label non-inferiority trial design comparing cefiderocol and standard of care antibiotics. Eligible participants will be adult inpatients who are diagnosed with a bloodstream infection with a Gram-negative organism on the basis of a positive blood culture result where the acquisition meets the definition for healthcare-associated or hospital-acquired. It will compare cefiderocol with the current standard of care (SOC) antibiotic regimen according to the patient’s treating clinician. Eligible participants will be randomised 1:1 to cefiderocol or SOC and receive 5–14 days of antibiotic therapy. Trial recruitment will occur in at least 20 sites in ten countries (Australia, Malaysia, Singapore, Thailand, Turkey and Greece). The sample size has been derived from an estimated 14 day, all-cause mortality rate of 10% in the control group, and a non-inferiority margin of 10% difference in the two groups. A minimum of 284 patients are required in total to achieve 80% power with a two-sided alpha level of 0.05. Data describing demographic information, risk factors, concomitant antibiotics, illness scores, microbiology, multidrug-resistant organism screening, discharge and mortality will be collected. Discussion With increasing antimicrobial resistance, there is a need for the development of new antibiotics with broad activity against Gram-negative pathogens such as cefiderocol. By selecting a population at risk for multi-drug-resistant pathogens and commencing study treatment early in the clinical illness (within 48 h of index blood culture) the trial hopes to provide guidance to clinicians of the efficacy of this novel agent. Trial registration The GAME CHANGER trial is registered under the US National Institute of Health ClinicalTrials.gov register, reference number NCT03869437 . Registered on March 11, 2019.

Urinary tract infection (UTI) causes significant renal damage and disease severity is compounded by antimicrobial resistance (AMR) and other comorbidities in the patient. Blood group antigens secreted in body fluids (secretor status) are known to play a role in bacterial adhesion and we studied its influence on AMR in UTI. A total of 2758 patients with UTI were studied with urine culture, qualitative and semiquantitative urine microscopy, serum creatinine and secretor status in saliva samples by adsorption-inhibition method. Of these, AMR from 300 patients with E. coli infection were assessed as per CLSI 2019 guidelines and extended-spectrum beta-lactamase (ESBL) genes (bla TEM, bla CTX-M, bla SHV) and NDM1 genes were studied using TaqMan probes in Real-time polymerase chain reaction. Patients with UTI were followed up for two weeks. Female patients had higher predilection (57%) for E. coli infection while patients with diabetes or non-secretors had none. In our study, ESBL producers were seen in 62% of the E. coli isolates and fosfomycin had 100% susceptibility. Non-secretors were significantly associated with acute kidney injury (AKI), AMR and ESBL genes. Multidrug-resistance (MDR) was noted in 127/160 (79.4%) ESBL and 17/18 (94%) NDM1 gene encoding strains. Quantitative urine microscopy scoring predicted AKI both at presentation and at end of follow up. ESBL producers were common in our study population and non-secretors had a significant association with AMR genes. Urine microscopy scoring system may be a useful tool to predict AKI in patients with UTI.

Foodborne infections caused by Salmonella spp. are among the most common foodborne diseases in the world. We isolated a lytic phage against extended-spectrum beta-lactam producing S. Enteritidis strain PT1 derived from chicken carcass. Results from electronmicrography indicated that phiPT1 belonged to the family, Siphoviridae, in the order, Caudovirales. Phage phiPT1 was stable at temperatures from 4 °C to 60 °C and inactivated at 90 °C. phiPT1 retained a high titer from pH 4 to pH 10 for at least 1 h. Nevertheless, it displayed a significant decrease (p 0.05). The phage-antibiotic combination analysis (PAS) indicated that synergism was not detected at higher phiPT1 titer (1012 PFU/mL) with all tested antibiotics at all subinhibitory concentrations. However, synergistic activities were recorded at 0.25 × MIC of four tested antibiotics: cefixime, gentamicin, ciprofloxacin, and aztreonam in combination with phage at 104, 106 and 108 PFU/mL (ΣFIC ≤0.5). Synergism was detected for all antibiotics (0.1 × MIC) except meropenem and colistin in combination with phiPT1 at 104, 106 and 108 PFU/mL (ΣFIC ≤0.5). Synergism also displayed at the lowest concentrations of all antibiotics (0.01 MIC) in combination with phiPT1 at all titers except 1012 PFU/mL. Such characteristic features make phiPT1 to be a potential candidate for therapeutic uses.

Previous studies have shown high acquisition risks of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-E) among international travelers visiting antimicrobial resistance (AMR) hotspots. Although antibiotic use and travelers’ diarrhea have shown to influence the ESBL-E acquisition risk, it remains largely unknown whether successful colonization of ESBL-E during travel is associated with the composition, functional capacity and resilience of the traveler’s microbiome. The microbiome of pre- and post-travel fecal samples from 190 international travelers visiting Africa or Asia was profiled using whole metagenome shotgun sequencing. A metagenomics species concept approach was used to determine the microbial composition, population diversity and functional capacity before travel and how it is altered longitudinally. Eleven travelers were positive for ESBL-E before travel and removed from the analysis. Neither the microbial richness (Chao1), diversity (effective Shannon) and community structure (Bray–Curtis dissimilarity) in pretravel samples nor the longitudinal change of these metrics during travel were predictive for ESBL-E acquisition. A zero-inflated two-step beta-regression model was used to determine how the longitudinal change in both prevalence and abundance of each taxon was related to ESBL acquisition. There were detected increases in both the prevalence and abundance of Citrobacter freundii and two members of the genus Bacteroides, in association with remaining uncolonized by ESBL-E. These results highlight the potential of these individual microbes as a microbial consortium to prevent the acquisition of ESBL-E. The ability to alter a person’s colonization resistance to a bacterium could be key to intervention strategies that aim to minimize the spread of MDR bacteria.

Klebsiella pneumoniae is a common cause of health-care associated infections. The rise of antibiotic resistance and the ability to form biofilm among K. pneumoniae strains are two key factors associated with antibiotic treatment failure. The present study investigates the antibiofilm activity of 1,8-cineole against preformed biofilms of multidrug-resistant extended-spectrum β-lactamase-producing K. pneumoniae clinical isolates. To evaluate the antibiofilm activity, cellular viability was analyzed by colony-forming units counting and live/dead staining. In addition, biofilm biomass was evaluated by crystal violet and the biofilm matrix was stained with calcofluor white and observed by confocal laser scanning microscopy. A time- and concentration-dependent effect of the phytochemical over biofilm cell viability was observed revealing that 1% (v/v) 1,8-cineole during 1 h was the optimal treatment condition displaying a significant reduction of cell viability in the preformed biofilms (2.5–5.3 log cfu/cm2). Furthermore, confocal laser scanning microscopy after SYTO-9 and propidium iodide staining showed that 1,8-cineole was capable of killing bacteria throughout all layers of the biofilm. The compound also caused a biofilm disruption (30–62% biomass reduction determined by crystal violet staining) and a significant decrease in biofilm matrix density. Altogether, our results demonstrate that 1,8-cineole is a promising candidate as a novel antibiofilm agent against multidrug-resistant K. pneumoniae strains producing extended-spectrum β-lactamases, given its capability to disrupt the structure and to kill cells within the biofilm.

Background: Antimicrobial resistance (AMR) is one of the top 10 public health threats. One approach to tackling the AMR menace could involve expanding the range of AMR surveillance domains to include hospital wastewater (HWW), a domain that has largely been overlooked by researchers. Aim: To evaluate the occurrence of multidrug-resistant bacteria in hospital wastewater of the Korle Bu Teaching Hospital (KBTH). Methodology: This was a longitudinal study involving 288 HWW samples consecutively collected across 12 weeks from the pool of wastewater emanating from 2 critical care units of KBTH—The Child Health Unit and the Maternity Unit—on Mondays and Thursdays, each week. The samples were cultured for bacteria, which were identified using the Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) technique and subjected to antimicrobial susceptibility testing via the Kirby-Bauer method. Results: In total, 294 bacteria of 23 different types, all being Gram-negative, were isolated from the 288 samples. The predominant ones were Escherichia coli (30.6%, n = 90), Klebsiella pneumoniae (11.2%, n = 33), Citrobacter freundii (10.9%, n = 32), Alcaligenes faecalis (5.8%, n = 17), and Pseudomonas mendocina (5.4%, n = 16). The prevalence of multidrug resistance among the isolates was 55.4% (n = 163). Moreover, the prevalence of extended-spectrum beta-lactamase (ESBL) producers was 15.6% (n = 46). E. coli accounted for the most ESBL-producing organisms (28.9%, n = 26). Conclusion: The wastewater generated by the Maternity and Child Health Units of KBTH harbored a wide range of multidrug resistant bacteria, with a good proportion of these being ESBL producers, and the predominant one being E. coli. The study thus identifies the wastewater of KBTH as an important source of multidrug resistant organisms, and underscores the significance of appropriate treatment of wastewater of the hospital and other clinical, and related settings prior to its discharge. [ABSTRACT FROM AUTHOR]

Background: Antimicrobial resistance is associated with increased morbidity in secondary infections and is a global threat owning to the ubiquitous nature of resistance genes in the environment. Recent estimate put the deaths associated with bacterial antimicrobial resistance in 2019 at 4.95 million worldwide. Lymphatic filariasis (LF), a Neglected Tropical Disease (NTD), is associated with the poor living in the tropical regions of the world. LF patients are prone to developing acute dermatolymphangioadenitis (ADLA), a condition that puts them at risk of developing secondary bacterial infections due to skin peeling. ADLA particularly worsens the prognosis of patients leading to usage of antibiotics as a therapeutic intervention. This may result in inappropriate usage of antibiotics due to self-medication and non-compliance; exacerbating antimicrobial resistance in LF patients. In this perspective, we assessed the possibilities of antimicrobial resistance in LF patients. We focused on antibiotic usage, antibiotic resistance in Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa isolates and looked at genes (mecA and Extended-spectrum beta-lactamase [blaCTX-M, blaSHV and blaTEM]) coding for resistance in multi-drug resistant (MDR) bacterial isolates. Results: Of the sixty (60) participants, fifty-four (n = 54, 90%) were within 31–60 years of age, twenty (n = 20, 33.33%) were unemployed and thirty-eight (n = 38, 50.67%) had wounds aged (in months) seven (7) months and above. Amoxicillin (54%) and chloramphenicol (22%) were the most frequently used antibiotics for self-medication. Staphylococcus aureus isolates (n = 26) were mostly resistant to penicillin (n = 23, 88.46%) and least resistant to erythromycin (n = 2, 7.69%). Escherichia coli isolates (n = 5) were resistant to tetracycline (n = 5, 100%) and ampicillin (n = 5, 100%) but were sensitive to meropenem (n = 5, 100%). Pseudomonas aeruginosa isolates (n = 8) were most resistant to meropenem (n = 3, 37.50%) and to a lesser ciprofloxacin (n = 2, 25%), gentamicin (n = 2, 25%) and ceftazidime (n = 2, 25%). Multi-drug resistant methicillin resistant Staphylococcus aureus (MRSA), cephalosporin resistant Escherichia coli. and carbapenem resistant Pseudomonas aeruginosa were four (n = 4, 15.38%), two (n = 2, 40%) and two (n = 2, 25%) respectively. ESBL (blaCTX-M) and mecA genes were implicated in the resistance mechanism of Escherichia coli and MRSA, respectively. Conclusion: The findings show presence of MDR isolates from LF patients presenting with chronic wounds; thus, the need to prioritize resistance of MDR bacteria into treatment strategies optimizing morbidity management protocols. This could guide antibiotic selection for treating LF patients presenting with ADLA. [ABSTRACT FROM AUTHOR]

Background: Ventilator Associated Pneumonia (VAP) is one of the common nosocomial infections associated with high morbidity due to multidrug resistant pathogens. Rapid spread of resistance to broad spectrum beta-lactams in pathogenic strains causes antibiotics ineffectiveness and increased severity of illness. The CTX-M is the most dominant Extended Spectrum ß Lactamase (ESBL) among Enterobacteriaceae in many regions of the world. The aim of the study was to identify the occurrence of ESBL. Materials and methods: This prospective study was conducted in the Department of Microbiology, Shadan Institute of Medical Sciences Teaching Hospital and Research Center. A total of 300 consecutive urine samples were screened from patients with symptomatic UTI. Clean-catch midstream urine samples were collected in sterile disposable container and processed within one hour. Semi quantitative loop measuring 2.2 mm diameter with a holding capacity of 0.005 ml was employed to culture urine on CLED agar and MacConkey's agar. The inoculated plates were incubated over night at 37°C. Isolates in significant number (colony count ≥ 105 CFU/ml) was identified by standard procedures. Results: In our study high susceptibility of meropenem (74.3%) and imipenem (100%) was seen and least was Ciprofloxacin 41.4%. Double disc synergy test and Inhibitory potentiated disc diffusion Test screening test for ESBL production, 70 (43.7%) isolates were resistant to at least one of the three representative cephalosporin drugs. The highest resistance was observed with cefpodoxime (n=57; 40.1%) followed by ceftazidime (n=52; 37.9%) and cefotaxime (n=39; 28.4%). Out of the three cephalosporins tested in the study, ceftazidime was found to be the better antibiotic drug for the identification of ESBL production by both Double disc synergy test and Inhibitory potentiated disc diffusion Test. Conclusion: This study showed that the ESBL-positive isolates were more resistant to some first-line antibiotics, and this highlights the necessity to control and monitor the prescribed antibiotics used for empirical treatment for UTI patients. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance (AMR) is one of the top global health threats of the 21th century. Recent studies are increasingly reporting the rise in extended-spectrum beta-lactamases producing Enterobacteriaceae (ESBLs-Ent) in dairy cattle and humans in the USA. The causes of the increased prevalence of ESBLs-Ent infections in humans and commensal ESBLs-Ent in dairy cattle farms are mostly unknown. However, the extensive use of beta-lactam antibiotics, especially third-generation cephalosporins (3GCs) in dairy farms and human health, can be implicated as a major driver for the rise in ESBLs-Ent. The rise in ESBLs-Ent, particularly ESBLs-Escherichia coli and ESBLs-Klebsiella species in the USA dairy cattle is not only an animal health issue but also a serious public health concern. The ESBLs-E. coli and -Klebsiella spp. can be transmitted to humans through direct contact with carrier animals or indirectly through the food chain or via the environment. The USA Centers for Disease Control and Prevention reports also showed continuous increase in community-associated human infections caused by ESBLs-Ent. Some studies attributed the elevated prevalence of ESBLs-Ent infections in humans to the frequent use of 3GCs in dairy farms. However, the status of ESBLs-Ent in dairy cattle and their contribution to human infections caused by ESBLs-producing enteric bacteria in the USA is the subject of further study. The aims of this review are to give in-depth insights into the status of ESBL-Ent in the USA dairy farms and its implication for public health and to highlight some critical research gaps that need to be addressed. [ABSTRACT FROM AUTHOR]

Background and Objectives: Upper urinary tract infections (UTIs) that require in-patient care can be expensive. Comorbid conditions such as diabetes as well as UTI due to extended-spectrum beta-lactamase (ESBL) producing bacteria may affect costs. The quality of life of patients with this condition has not been described. Methods: This was a cost of illness study that prospectively evaluated patients admitted with upper UTI to a medical ward in a tertiary care hospital. Direct medical and nonmedical costs, indirect costs were collected to make the total cost per admission. Quality of life was assessed using the World Health Organization Quality of Life-BREF score. We also compared costs between those with and without diabetes or ESBL infection. Results: Between March 2016 and July 2017, 92 eligible patients were included in the study. The average age was 55.8 years; two thirds were diabetics. The mean overall cost of a single admission for upper UTI was INR.88, 330.2 (1370.4 USD). This was INR.96, 193.0 (1492.6 USD) and INR.1, 03,154.9 (1600.4 USD) among those with diabetes mellitus and ESBL infection, respectively. The cost was higher among those with diabetes and ESBL than those without; this difference reached statistically significance for the ESBL group. The quality of life was affected in all domains; the psychological being most affected among diabetics and ESBL infected. Conclusions: The mean total cost of admission for an upper UTI in a tertiary care hospital in South India was INR 88,330 (1370.4 USD). This is higher if the patient has diabetes or ESBL organism causing the UTI. Quality of life is clearly reduced especially in the psychological domain. [ABSTRACT FROM AUTHOR]

Background: Cefoxitin is active against some extended-spectrum beta-lactamase-producing Enterobacterales (ESBL-PE), but has not been evaluated so far in the intensive care unit (ICU) settings. Data upon its pharmacokinetics (PK), tolerance and efficacy in critical conditions are scanty. We performed a retrospective single-center study in a university hospital medical ICU, in subjects presenting with cefoxitin-susceptible ESBL-PE infection and treated with cefoxitin. The primary aim was to determine cefoxitin PK. Secondary endpoints were efficacy, tolerance, and emergence of cephamycin-resistance. Results: Forty-one patients were included in this study, mainly with ESBL-PE pneumonia (35 patients, 85%). Cefoxitin was administered during a median [interquartile range (IQR)] duration of 5 [4–7] days. Cefoxitin serum concentrations strongly depended on renal function. Target serum concentration (> 5 × minimum inhibitory concentration (MIC) 24 h after cefoxitin onset was obtained in 34 patients (83%), using a median [IQR] daily dose of 6 [6–6] g with continuous administration. The standard dosage of 6 g/24 h was not sufficient to achieve the PK/PD target serum concentration for MIC up to 4–8 mg/L, except in patients with severe renal impairment and those treated with renal replacement therapy. Treatment failure occurred in 26 cases (63%), among whom 12 patients (29%) died, 13 patients (32%) were switched to alternative antibiotic therapy and 11 patients (27%) presented with relapse of infection with the same ESBL-PE. Serious adverse events attributed to cefoxitin occurred in 7 patients (17%). Acquisition of cephamycin-resistance with the same Enterobacterales was identified in 13 patients (32%), and was associated with underdosage. Conclusion: Continuous administration of large doses of cefoxitin appears necessary to achieve the PK/PD target in patients with normal renal function. Renal status, MIC determination and therapeutic drug monitoring may be useful for treatment individualization in this setting. The treatment failure rate was 63%. The cefoxitin safety profile was favorable, but we observed a high rate of cephamycin-resistance emergence. [ABSTRACT FROM AUTHOR]

The incidence of urinary tract infections (UTIs) caused by extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E. coli) bacteria has become a global problem and has increased in recent years. The purpose of this study was to analyze the risk factors for the incidence of UTI in inpatients at Dr. Soetomo General Academic Hospital, Surabaya. This research was an analytic descriptive study. Specimens were sent to the Clinical Microbiology Laboratory using sterile containers and processed according to standard laboratory procedures. It resulted in ESBL-producing E. coli that were used as a case group and non ESBL-producing E. coli as a control group. The identification and testing of antibiotic susceptibility were carried out using the BD Phoenix™ Automated Microbiology System. Ninety-four bacterial isolates were collected, consisting of 54 (57.4%) ESBL-producing E. coli bacteria and 40 (42.6%) non ESBL-producing E. coli bacteria. Recurrent UTIs (OR = 4.31; ρ =0.002; 95% CI = 1.68-11.04) and catheter use (ρ = 0.049; OR = 4.250; 95% CI = 1.050-17.210) were used as independent risk factors caused by ESBL-producing E. coli bacteria. Recurrent UTIs and catheter use were dependent risk factors caused by ESBL-producing E. coli bacteria. [ABSTRACT FROM AUTHOR]

Introduction: The incidence of orthopaedic implantation and implantation associated infections were correspondingly increasing among the elderly and trauma patients. The resistance among the pathogens pose a unique challenge to the clinicians in the management of the infection. Extended-Spectrum Beta-Lactamase (ESBL), AmpC beta-lactamases (AmpC), Metallo Beta-Lactamases (MBL) and Methicillin-Resistant Staphylococcus aureus (MRSA) producing bacterial pathogens are responsible for a high rate of relapsing infections and outbreaks of nosocomial infections. Aim: To focus on accounting the incidence of ESBL, AmpC, MBL and MRSA and its antibiogram for effective management of implant associated infection. Materials and Methods: The present study was a prospective observational study which was conducted using various samples like pus, wound swab, serous discharge. These samples were collected under aseptic precautions in sterile containers for the period of one year (February 2015-February 2016). A total of 200 samples were inoculated on Blood agar and McConkey agar and identified. The isolates were tested for detection of ESBL, AmpC, MBL and MRSA as per standard protocol. The statistical analysis was performed using Statistical Package for the Social Sciences (SPSS) software version 20.0 Chicago, USA. Results: Most commonly used implants were intramedullary interlocking nail 138 (65.7%) followed by dynamic compression plates 23 (11.0%) and anatomical plates 18 (8.6%). Of the total of 200 samples, 190 (95%) samples yielded monomicrobial isolates and 10 (5%) samples yielded polymicrobial isolate. gram negative isolates 108 (51.4%) were marginally higher than gram positive isolates 102 (48.6%). The predominant isolate was S. aureus 81 (38.5%) followed by Klebsiella spp. 28 (13.3%) and Pseudomonas aeruginosa 21 (10.0%). In gram negative isolates, 76 (70.4%) were ESBL and/or AmpC producers. Of which, 48 (44.4%) were ESBL and AmpC co-producers. There were no MBL producing isolates. In gram positive isolates, majority were S. aureus 81 (79.4%) followed by Coagulase-negative Staphylococcus 21 (20.6%). Out of 81 S. aureus isolates, 46 (56.8%) were MRSA. Out of 21 CoNS, majority 17 (81%) isolates were methicillin-resistant. Implant failures were observed in 28 (14%) cases. Out of 28, majority 12 (42.8%) of the implant failures were MRSA. Conclusion: High rates of ESBL, AmpC and MRSA infections associated with implant surgeries indicate the necessity to formulate antibiotic policies and control measures. ESBL and AmpC producing strains were found to show higher rates of resistance to the various class of antibiotics when compared to non-ESBL and non-AmpC producers. MRSA isolates were found to show higher rates of resistance to various classes of antibiotics when compared to MSSA.

Abstract Background The prevalence of infections caused by extended-spectrum beta-lactamase producing Escherichia coli (ESBL-E. coli) is increasing worldwide, but the setting in which this increase is occurring is not well defined. We compared trends and risk factors for ESBL-E. coli bacteriuria in community vs healthcare settings. Methods We collected electronic health record data on all patients with E. coli isolated from urine cultures in a safety-net public healthcare system from January 2014 to March 2020. All analyses were stratified by healthcare-onset/associated (bacteriuria diagnosed > 48 h after hospital admission or in an individual hospitalized in the past 90 days or in a skilled nursing facility resident, N = 1277) or community-onset bacteriuria (bacteriuria diagnosed 65 (RR 1.88, CI 1.17, 3.05), male gender (RR 2.12, CI 1.65, 2.73), and Latinx race/ethnicity (RR 1.52, CI 0.99, 2.33) were associated with community-onset ESBL-E. coli. Only male gender (RR 1.53, CI 1.03, 2.26) was associated with healthcare-onset/associated ESBL-E. coli. Conclusions ESBL-E. coli bacteriuria frequency increased at a faster rate in healthcare-associated settings than in the community between 2014 and 2020. Male gender was associated with ESBL-E. coli bacteriuria in both settings, but additional risks—age > 65 and Latinx race/ethnicity—were observed only in the community.

The infection with extended-spectrum beta-lactamase-producing Escherichia coli is associated with higher mortality, longer length of hospital-stay and increased costs compared to infection with antibiotic-susceptible E. coli. Here, the draft genome of ESBL-producing E. coli circulating at local hospital is reported. The strain was detected as containing the genes of antibiotic resistance TEM, CTX-M-1, and CTX-M-9. The 5,136,548-bp genome, with a GC content of 50.59%, comprised 4987 protein-coding genes, four ribosomal RNA, and 66 transfer RNA. The ResFinder was successfully predicted fourteen antimicrobial genes in the E. coli INF191/17/A genome. Sequence data has been deposited in the GenBank database under the accession number JAIEXV000000000. The BioProject ID in the GenBank database is PRJNA752944. The raw data was sequenced using Ilumina MiSeq and submitted to the NCBI SRA database (SRX11797310), which is publicly available.

Abstract Background Highly empiric use of carbapenem in pyogenic liver abscess (PLA) is widespread problem. However, few studies have examined the association between blood culture and carbapenem use in patients with PLA in China. Thus, we conducted this observational study. Methods The data of patients diagnosed with PLA at two comprehensive tertiary care centers from 2014 to 2020 were retrospectively collected. Demographic and clinical data were analyzed, and univariate and multivariate analyses were performed to investigate the association between blood culture and carbapenem use. Subgroup analysis was conducted to explore whether the effect is different in sepsis. Results Blood culture was performed in 110 (46.0%) patients, of whom 44 (40.0%) patients had positive results for bacterial culture. Extended-spectrum beta-lactamase (ESBL)-positive blood culture isolates were detected in 8 (7.3%) patients. The positivity rate of blood culture in sepsis was higher than in non-sepsis (58.1% vs. 32.9%, P = 0.015). Fewer patients who had a blood culture received carbapenem treatment in comparison to patients without blood culture (19.1% vs. 31.8%, P = 0.026). Multivariate analysis showed that blood culture was independently associated with less carbapenem exposure (adjusted odds ratio [OR] = 0.33, 95% confidence interval [CI]: 0.16–0.68, P = 0.003), and this effect remained significant in the sepsis subgroup (adjusted OR = 0.17, 95% CI: 0.05–0.53, P = 0.002). Conclusion Blood culture had a high positivity rate and was associated with less carbapenem use in PLA, especially those who developed sepsis. More attention should be paid to performing early blood culture and less carbapenem use in PLA.

Urinary tract infections (UTIs) are one of the most frequent bacterial infections in the world, both in the hospital and community settings. Uropathogenic Escherichia coli (UPEC) are the predominant etiological agents causing UTIs. Extended-spectrum beta-lactamase (ESBL) production is a prominent mechanism of resistance that hinders the antimicrobial treatment of UTIs caused by UPEC and poses a substantial danger to the arsenal of antibiotics now in use. As bacteria have several methods to counteract the effects of antibiotics, identifying new potential drug targets may help in the design of new antimicrobial agents, and in the control of the rising trend of antimicrobial resistance (AMR). The public availability of the entire genome sequences of humans and many disease-causing organisms has accelerated the hunt for viable therapeutic targets. Using a unique, hierarchical, in silico technique using computational tools, we discovered and described potential therapeutic drug targets against the ESBL-producing UPEC strain NA114. Three different sets of proteins (chokepoint, virulence, and resistance genes) were explored in phase 1. In phase 2, proteins shortlisted from phase 1 were analyzed for their essentiality, non-homology to the human genome, and gut flora. In phase 3, the further shortlisted putative drug targets were qualitatively characterized, including their subcellular location, broad-spectrum potential, and druggability evaluations. We found seven distinct targets for the pathogen that showed no similarity to the human proteome. Thus, possibilities for cross-reactivity between a target-specific antibacterial and human proteins were minimized. The subcellular locations of two targets, ECNA114_0085 and ECNA114_1060, were predicted as cytoplasmic and periplasmic, respectively. These proteins play an important role in bacterial peptidoglycan biosynthesis and inositol phosphate metabolism, and can be used in the design of drugs against these bacteria. Inhibition of these proteins will be helpful to combat infections caused by MDR UPEC.

Due to the rapid spread of CTX-M type ESBLs, the rate of resistance to third-generation cephalosporin has increased among Gram-negative bacteria, especially in Escherichia coli, and there is a need to find ways to re-sensitize ESBL E. coli to cephalosporin treatment. A previous study showed that genes involved in protein synthesis were significantly up-regulated in the presence of subinhibitory concentration of cefotaxime (CTX) in a CTX-M-1-producing E. coli. In this study, the interaction between CTX and gentamicin (GEN), targeting protein synthesis, was evaluated in MG1655/pTF2, and the MIC of CTX was strongly reduced (128-fold) in the presence of this combnation therapy. Since the underlying mechanism behind this synergy is not known, we constructed a saturated transposon mutant library in MG1655/pTF2::blaCTX-M-1 containing 315,925 unique transposon insertions to measure mutant depletion upon exposure to CTX, GEN, and combination treatment of CTX and GEN by Transposon Directed Insertion-site Sequencing (TraDIS). We identified 57 genes that were depleted (log2FC ≤ −2 and with q.value ≤ 0.01) during exposure to CTX, 18 for GEN, and 31 for combination treatment of CTX and GEN. For validation, we deleted eight genes that were either uniquely identified in combination treatment, overlapped with monotherapy of GEN, or were shared between combination treatment and monotherapy with CTX and GEN. Of these genes, we found that the inactivation of dnaK, mnmA, rsgA, and ybeD increased the efficacy of both CTX and GEN treatment, the inactivation of cpxR and yafN increased the efficacy of only CTX, and the inactivation of mnmA, rsgA, and ybeD resulted in increased synergy between CTX and GEN. Thus, the study points to putative targets for helper drugs that can restore susceptibility to these important drugs, and it indicates that genes involved in protein synthesis are essential for the synergy between these two drugs. In summary, the study identified mutants that sensitize ESBL-producing E. coli to CTX and a combination of CTX and GEN, and it increased our understanding of the mechanism behind synergy between β-lactam and aminoglycoside drugs. This forms a framework for developing new strategies to combat infections caused by resistant bacteria.