OXA Genes Research Articles

The emergence of plasmid-encoded oxacillinase and carbapenemase among uropathogenic Escherichia coli (UPEC) isolated from hospitalized patients in the North of Iran

Carbapenemase enzyme production is responsible for resistance to carbapenem among Gram-negative bacteria. This study aimed to detect common carbapenemase and oxacilinase genes among uropathogenic E. coli (UPEC) isolated from hospitalized patients in Rasht, north of Iran. In the present study, from 2000 urine samples, 263 UPEC strains were isolated from inpatients with urinary tract infections (UTI) in 2020. The Kirby-Bauer disk diffusion susceptibility test was used to determine the sensitivity or resistance of isolates to antimicrobial compounds. The double-disk test confirmed extended-spectrum β lactamase (ESBL) production phenotypically, and the presence and distribution of genes encoding carbapenemase and oxacilinase were investigated using polymerase chain reaction (PCR). Based on the findings, 13/263 isolates (8 ESBL and five non-ESBL) showed a non-susceptible phenotype to at least one of the studied carbapenem group antibiotics, and 121 (46%) isolates were ESBL-producers. PCR for oxacilinase and carbapenemase genes was done on all 126 isolates, including ESBL-positive and carbapenem-resistant strains, in which 10 (7.9%) and 25 (19.8%) isolates harbored OXA-1 and IMP genes, respectively. Also, OXA-2, OXA-10, OXA-48, VIM, and NDM genes were not found in any studied isolates. IMP and OXA-1 genes among carbapenemase-producing isolates indicate the possible spread of antibiotic-resistant strains. Hence, identification and control of ESBL and carbapenemase-producing strains, although with almost low frequency due to plasmid genes encoding carbapenemase, is essential for infection control.

High prevalence of extensively drug resistant and extended spectrum beta lactamases (ESBLs) producing uropathogenic Escherichia coli isolated from Faisalabad, Pakistan.

Urinary tract infections (UTIs) are predominantly caused by uropathogenic Escherichia coli (E. coli). There is rapid increase in antimicrobial resistance in UTIs, also declared as a serious health threat by World Health Organization (WHO). Present study was designed to investigate the antimicrobial resistance status with specific focus on ESBLs and carbapenemases in local uropathogenic E. coli (UPEC) isolates. E. coli isolates were characterized from patients of all ages visiting diagnostic laboratories for urine examination. Demographic data was also recorded for each patient. Antibiograms were developed to observe antibiotic resistance in UPEC using Kirby Bauer disc diffusion technique. Double Disc Synergy test (DDST) was used for phenotypic ESBL test. ESBLs and carbapenemases genes were detected in UPEC using PCR. The PCR results were confirmed by sequencing. The UPEC isolates under study exhibited 78%, 77%, 74%, 72% and 55% resistance against cefotaxime, amoxicillin, erythromycin, ceftriaxone and cefixime, respectively. Resistance against colistin and meropenem was observed in 64% and 34% isolates, respectively. Phenotypic DDST identified 48% isolates as ESBLs producers. Genotypic characterization identified 70%, 74.4% and 49% prevalence of CTXM-1, TEM-1 and CTXM-15 genes respectively. One isolate was observed exhibiting co-existence of all ESBL genes. TEM-1 + CTXM-1 and TEM-1 + CTXM-1 + CTXM-15 + OXA-1 gene patterns were dominant among ESBLs. For carbapenem-resistance, 14% isolates indicated the presence of KPC whereas GES and VIM was detected in 7% and 3.4% isolates, respectively. In conclusion, our results present a high prevalence of extensively drug resistant UPEC isolates with a considerable percentage of ESBL producers. These findings propose the need of continuous surveillance for antimicrobial resistance and targeted antimicrobial therapy.

The European Union Summary Report on Antimicrobial Resistance in zoonotic and indicator bacteria from humans, animals and food in 2020/2021.

Antimicrobial resistance (AMR) data on zoonotic and indicator bacteria from humans, animals and food are collected annually by the EU Member States (MSs) and reporting countries, jointly analysed by EFSA and ECDC and presented in a yearly EU Summary Report. This report provides an overview of the main findings of the 2020-2021 harmonised AMR monitoring in Salmonella spp., Campylobacter jejuni and C. coli in humans and food-producing animals (broilers, laying hens and turkeys, fattening pigs and bovines under 1year of age) and relevant meat thereof. For animals and meat thereof, indicator E. coli data on the occurrence of AMR and presumptive Extended spectrum β-lactamases (ESBL)-/AmpC β-lactamases (AmpC)-/carbapenemases (CP)-producers, as well as the occurrence of methicillin-resistant Staphylococcus aureus are also analysed. In 2021, MSs submitted for the first time AMR data on E. coli isolates from meat sampled at border control posts. Where available, monitoring data from humans, food-producing animals and meat thereof were combined and compared at the EU level, with emphasis on multidrug resistance, complete susceptibility and combined resistance patterns to selected and critically important antimicrobials, as well as Salmonella and E. coli isolates exhibiting ESBL-/AmpC-/carbapenemase phenotypes. Resistance was frequently found to commonly used antimicrobials in Salmonella spp. and Campylobacter isolates from humans and animals. Combined resistance to critically important antimicrobials was mainly observed at low levels except in some Salmonella serotypes and in C. coli in some countries. The reporting of a number of CP-producing E.coli isolates (harbouring bla OXA-48, bla OXA-181, and bla NDM-5 genes) in pigs, bovines and meat thereof by a limited number of MSs (4) in 2021, requests a thorough follow-up. The temporal trend analyses in both key outcome indicators (rate of complete susceptibility and prevalence of ESBL-/AmpC- producers) showed that encouraging progress have been registered in reducing AMR in food-producing animals in several EU MSs over the last years.

Prevalence of Carbapenem-Resistant Enterobacteriaceae in Western Saudi Arabia and Increasing Trends in the Antimicrobial Resistance of Enterobacteriaceae.

The aim of the study is to estimate the prevalence rate of carbapenem-resistant Enterobacteriaceae (CRE) and to determine the types of carbapenemase genes present in patients admitted to King Abdulaziz Medical City (KAMC-J) and King Abdulaziz University Hospital (KAUH), both in Jeddah, Saudi Arabia. A total of 180 isolates were analyzed which were included on the basis of retrospective chart review of patients from KAMC-J and KAUH between 1st April 2017 to 30th March 2019. The prevalence of carbapenemase genes( blaIMP, blaVIM, blaKPC, blaNDM-1, and blaOXA-48)was evaluated by Xpert® Carba-R (Cepheid, Sunnyvale, CA, USA). We assessed the CRE prevalence and described their susceptibility to antimicrobial agents based on antibiogram reports. Results:Klebsiella pneumoniaeshowed a higher frequency of bla OXA-48 (79%) than bla NDM (11.7%) genes (p=0.007). The CRE prevalence in KAUH was 8% in 2017 and increased to 13% in 2018. In KAMC-J, the prevalence was 57% in 2018 and 61% in 2019. K. pneumoniae was found to be the most frequently isolated causative organism followed byEscherichia coli .The bla OXA-48 (76.1%) gene was predominantamong overall isolates followed by bla NDM (13.9%); both genes coexisted in 6.1% of the isolates. During the study period, the prevalence of CRE considerably rose in the two tertiary care institutions from western Saudi Arabia. In the CRE isolates, bla OXA-48was discovered to be the most common gene. We recommend an antimicrobial resistance surveillance system to detect the emergence of resistant genes through use of new rapid diagnostic tests and monitor antimicrobial use in order to improve clinical outcomes of CRE infections given the severity of infection associated with the CRE isolates as well as the limited treatment options available.

Colonization and Healthcare-Associated Infection of Carbapenem-Resistant Enterobacteriaceae, Data from Polish Hospital with High Incidence of Carbapenem-Resistant Enterobacteriaceae, Does Active Target Screening Matter?

The objective of the study was to analyse the incidence of carbapenem-resistant Enterobacteriaceae (CRE) at a provincial hospital from 2019-2021. Multiplex PCR was used to detect the presence of carbapenemase genes. There were 399 cases of CRE detected in total in the analysed period, including 104 healthcare-associated infections. Out of the isolated CRE, 97.7% were Klebsiella pneumoniae with OXA-48 or KPC genes. Overall, among the identified CRE genes, the most frequently present genes were the ones mediating oxacillinase OXA-48 (71%) and KPC (26%), and significantly less often New Delhi NDM metallo-β-lactamase (2.5%). Moreover, two isolates produced two carbapenemases, i.e., OXA-48 and KPC. The conducted research demonstrates that there is a constant need for continuous monitoring of the occurrence of CRE strains and the hospital antibiotic policy, as well as the implementation of procedures to prevent CRE transmission by medical personnel and hospital support staff.

OprL Gene Sequencing, Resistance Patterns, Virulence Genes, Quorum Sensing and Antibiotic Resistance Genes of XDR Pseudomonas aeruginosa Isolated from Broiler Chickens.

Pseudomonas aeruginosa is incriminated in septicemia, significant economic losses in the poultry production sector, and severe respiratory infections in humans. This study aimed to investigate the occurrence, oprL sequencing, antimicrobial resistance patterns, virulence-determinant, Quorum sensing, and antibiotic resistance genes of P. aeruginosa retrieved from broiler chickens. Two hundred samples were collected from 120 broiler chickens from broiler farms at Ismailia Governorate, Egypt. Consequently, the bacteriological examination was conducted and the obtained P. aeruginosa strains were tested for oprL gene sequencing, antibiogram, and PCR screening of virulence, Quorum sensing, and antibiotic resistance genes. The overall prevalence of P. aeruginosa in the examined birds was 28.3%. The oprL gene sequence analysis underlined that the tested strain expressed a notable genetic identity with various P. aeruginosa strains isolated from different geographical areas in the USA, India, China, Chile, and Ghana. PCR evidenced that the obtained P. aeruginosa strains, carrying virulence-related genes: oprL, toxA, aprA, phzM, and exoS in a prevalence of 100%, 100%, 42.5%, 33.3%, and 25.9%, respectively. Moreover, the recovered P. aeruginosa strains possessed the Quorum sensing genes: lasI, lasR, rhlI, and rhlR in a prevalence of 85.2%, 85.2%, 81.5%, and 81.5%, respectively. Furthermore, 40.7% of the isolated P. aeruginosa were XDR to seven antimicrobial classes, possessing sul1, bla TEM, tetA, bla CTX-M, bla OXA-1, and aadA1 genes. As we can tell,thisisthefirstreportemphasizing theevolutionofXDRP. aeruginosastrainsfrom broiler chicken in Egypt, which is supposed to be a serious threat to public health.The emerging XDR P. aeruginosa in poultry frequently harbored the oprL, toxA, and aprA virulence genes, the lasI, lasR, rhlI, and rhlR Quorum sensing genes, and the sul1, bla TEM, tetA, bla CTXM, bla OXA-1, and aadA1 resistance genes.

Surveillance of antibiotic resistance in the northern zone of Senegal: Molecular characterization of beta-lactam resistance genes in Enterobacteriaceae (extended spectrum beta-lactamase, plasmid cephalosporinases, carbapenemases).

Introduction: With the increase in bacterial resistance to antibiotics, policies are being implemented on a national scale. These resistances are multiple, especially in enterobacteria, and represent a real public health problem due to the difficulty of their treatment. The objective of this work was to identify ESBL, cephalosporinase and carbapenemase producing enterobacteria in laboratories in the northern part of Senegal and to compare the mechanisms involved. Materials and methods: This is a retrospective study conducted at the Saint-Louis Regional Hospital and the Microbiology Unit of the Pasteur Institute of Dakar. Bacterial strains were collected between January and June 2021 in eight bacteriology laboratories in the regions of Saint-Louis, Louga and Matam. Bacterial identification was done by studying morphological, cultural and biochemical characteristics using an API® 20 E gallery and the MALDI-TOFF technique. The antibiogram was performed by the disc diffusion method according to the CA-SFM version 2021 procedure including the search for specific resistances (ESBL). The ESBL genes (CTX, TEM, OXA, and SHV) were identified by polymerase chain reaction (PCR) with specific primers and then analyzed by gel electrophoresis. Results: Forty-five strains of Enterobacteriaceae were ESBL producers and each had at least one beta-lactam resistance gene. The CTX-M gene was found in all strains, followed by the OXA (64.44%), TEM (53.33%) and SHV (20%) genes. A simultaneous presence of these 4 genes was noted in 6 isolates (13.33%). Apart from resistance to ertapenem, all strains were sensitive to imipenem. For quinolones, almost all strains (93%) were resistant. Conclusion: These results show the need to reinforce the surveillance of ESBL-producing Enterobacteriaceae in the Senegal area. These important data should help to orientate strategies for the reduction of MRBs and their dissemination in the community.

MLST typing of Acinetobacter baumannii in a hospital outbreak

Background and objective: Carbapenem-resistant Acinetobacter baumannii (CRA) is a WHO-designated priority antibiotic-resistant pathogen. In this study, we performed Multi-Locus Sequence Typing (MLST) for the clinical and environmental CRA isolates recovered from an investigation of a hospital CRA outbreak. Methods: CRA was recovered from 58 patients and multiple environmental sites. Twenty-four clinical and 11 environmental isolates were selected for MLST. MALDI-TOF and OXA-51 gene PCR were used for identification of Acinetobacter baumannii. MLST was done by Sanger sequencing. Fisher’s exact test was used for analysis of categorical variables. Results: Extensive patient screening and environmental sampling identified the hospital laundry as the likely source of CRA outbreak across multiple wards. CRA isolates of ST345 from hospital laundry cooler machine was identical to that of 11 patients across different wards. CRA isolates of ST1145 from three laundry linen racks were identical to that of seven patients across different wards. CRA isolates of ST2207 found in another linen rack were identical to that in three other patients across different wards. Antimicrobial susceptibility patterns of CRA isolates could not accurately differentiate between different ST. Conclusion: MLST result demonstrated the link between contamination of laundry cooler and environment and the hospital polyclonal CRA outbreak.

Detection and homology analysis of carbapenem resistant Acinetobacter baumannii resistance gene.

To explore the carrying status and homology of carbapenem resistant Acinetobacter baumannii (CRAB) in our hospital. From January 2015 to December 2017, 52 strains of acinetobacter baumannii isolated from the bacteria room of the clinical laboratory of Baogang hospital in Inner Mongolia were selected as the research object. K-B disk diffusion method and Vitek-2 were used to determine the drug sensitivity of Acinetobacter baumannii. The drug resistance gene was detected by polymerase chain reaction (PCR) and its homology was analyzed by pulsed field gel electrophoresis (PFGE). Except for Cefoperazone/sulbactam, other antibiotics were resistant to ab. The detection rate of drug resistance gene class C β-lactamases (ADC) was 100%, and the higher detection rates of other drug resistance genes were class D β-lactamases (OXA)-51 (36 strains, 90.0%),disinfectant gene qacE△1-sull (32 strains, 80.0%), and klebsiella pneumoniae carbapenemase (KPC) gene was not detected. 2-8 drug resistance genes were detected in each CRAB strain, and the strains with 6 drug resistance genes were the most (15 strains, 37.5%); Among the detected drug-resistant gene combinations, ADC+OXA-23 + OXA-51 gene was detected at the same time (29 strains, 72.5%), followed by ADC+ intl1 + qacE △ 1-sull gene (26 strains, 65.0%), ADC + qacE △ 1-sull + ant (3 '') -i gene (19 strains, 47.5%), and 11 strains (27.5%). There were 19 different types in PFGE homology test, each type was 1-9 strains, including 9 strains of A5 type and 8 strains of A18 type, mainly from intensive care unit. CRAB in the hospital is highly resistant to common clinical antibiotics. OXA-23 and OXA-51 genes are most likely to be the main factors causing drug resistance of Acinetobacter baumannii in the hospital. Homology analysis showed that there was CRAB nosocomial infection transmission in different wards of the hospital.

Susceptibility profile of bla OXA-23 and metallo-β-lactamases co-harbouring isolates of carbapenem resistant Acinetobacter baumannii (CRAB) against standard drugs and combinations.

The rapid emergence of carbapenem resistant Acinetobacter baumannii (CRAB) has resulted in an alarming situation worldwide. Realizing the dearth of literature on susceptibility of CRAB in genetic context in the developing region, this study was performed to determine the susceptibility profile against standard drugs/combinations and the association of in-vitro drug synergy with the prevalent molecular determinants. A total of 356 clinical isolates of A. baumannii were studied. Confirmation of the isolates was done by amplifying recA and ITS region genes. Susceptibility against standard drugs was tested by Kirby Bauer disc diffusion. Minimum inhibitory concentration (MIC), MIC50 and MIC90 values against imipenem, meropenem, doripenem, ampicillin/sulbactam, minocycline, amikacin, polymyxin B, colistin and tigecycline was tested as per guidelines. Genes encoding enzymes classes A (bla GES, bla IMI/NMC-A, bla SME, bla KPC), B (bla IMP, bla VIM, bla NDM) and D (bla OXA-51, bla OXA-23 and bla OXA-58) were detected by multiplex polymerase chain reaction. Synergy against meropenem-sulbactam and meropenem-colistin combinations was done by checkerboard MIC method. Correlation of drug synergy and carbapenemase encoding genes was statistically analyzed. Of the total, resistance above 90% was noted against gentamicin, ciprofloxacin, levofloxacin, ceftazidime, cefepime, ceftriaxone, cotrimoxazole and piperacillin/tazobactam. By MIC, resistance rates from highest to lowest was seen against imipenem 89.04% (n=317), amikacin 80.33% (n=286), meropenem 79.49% (n=283), doripenem 77.80% (n=277), ampicillin/sulbactam 71.62% (n=255), tigecycline 55.61% (n=198), minocycline 14.04% (n=50), polymyxin B 10.11% (n=36), and colistin 2.52% (n=9). CRAB was 317 (89.04%), 81.46% (n=290) were multidrug resistant and 13.48% (n=48) were extensively drug resistant. All the CRAB isolates harboured bla OXA-51 gene (100%) and 94% (n=298) bla OXA-23 gene. The bla IMP gene was most prevalent 70.03% (n=222) followed by bla NDM, 59.62% (n=189). Majority (87.69%, 278) were co-producers of classes D and B carbapenemases, bla OXA-23 with bla IMP and bla NDM being the commonest. Synergy with meropenem-sulbactam and meropenem-colistin was 47% and 57% respectively. Reduced synergy (p= <0.0001) was noted for those harbouring bla OXA-51+blaOXA-23with bla NDM gene alone or co-producers. Presence of bla NDM gene was a significant cause of synergy loss in meropenem-sulbactam and meropenem-colistin. In bla NDM endemic regions, tigecycline, minocycline and polymyxins could be viable options against CRAB isolates with more than one carbapenemase encoding genes.

Trends in carbapenem resistance in Pre-COVID and COVID times in a tertiary care hospital in North India

BackgroundCarbapenem resistance is endemic in the Indian sub-continent. In this study, carbapenem resistance rates and the prevalence of different carbapenemases were determined in Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa during two periods; Pre-COVID (August to October 2019) and COVID (January to February 2021) in a north-Indian tertiary care hospital.MethodsDetails of patient demographics and clinical condition was collated from the Hospital Information System and detection of carbapenemases NDM, OXA-48, VIM, IMP and KPC was done by Polymerase chain reaction (PCR) in 152 and 138 non-consecutive carbapenem resistant isolates during the two study periods respectively. Conjugation assay and sequencing of NDM and OXA-48 gene was done on a few selected isolates.ResultsAs compared to Pre-COVID period, co-morbidities and the mortality rates were higher in patients harbouring carbapenem resistant organisms during the COVID period. The overall carbapenem resistance rate for all the four organisms increased from 23 to 41% between the two periods of study; with Pseudomonas aeruginosa and Klebsiella pneumoniae showing significant increase (p < 0.05). OXA-48, NDM and co-expression of NDM and OXA-48 were the most common genotypes detected. NDM-5 and OXA-232 were most common variants of NDM and OXA-48 family respectively during both the study periods.ConclusionHigher rate of carbapenem resistance in COVID times could be attributed to increase in number of patients with co-morbidities. However, genetic elements of carbapenem resistance largely remained the same in the two time periods.

Antibiogram, Genomic and Phylogeny of Stool and Seafood Isolates from Some Cholera-Prone Coastal Communities in Rivers State, Nigeria

Cholera is a significant public health threat across the globe, especially in coastal regions with poor water supply. This study was carried out to determine the antibiogram, genomic, and phylogeny of stool and seafood isolates from some cholera-prone coastal communities in Rivers State, Nigeria. A total of 400 stool samples and 42 different seafood were aseptically collected and examined using standard microbiology and molecular techniques. An antibiogram of isolates from seafood and stool samples was assayed. Genes for virulence, resistance, and relatedness of bacteria identified were also determined. The isolates from the stool and seafood were examined for susceptibility to some selected antibiotics. The findings showed the prevalence rate of cholera in the communities as follows: 16% in Kaa, 30% in Andoni, 4% in Ogu/Bolo, and 10% in Abua/Odual. The isolates from stool were susceptible to Ciprofloxacin and Gentamycin with a susceptibility rate of 94.12% each while 100% resistance was recorded against Amoxicillin-clavulanic acid, 94.12% against Amikacin and 88.24% against Colistin. For the sea foods, the isolates were susceptible to gentamycin and ciprofloxacin with a susceptibility rate of 91.43% and 82.86% respectively. Resistance was also recorded against Colistin (88.57%) and Azithromycin (82.86%). Testing the isolates for the presence of 16SrRNA genes showed that all were positive with 1500 bp 16SrRNA gene band size. TEM, OXA, SHV, and CTX-M resistant genes were detected whereas the virulence genes were TDH and AcrB. The phylogenetic analysis revealed isolates from seafood to be Aeromonas dhakensis, Vibrio parahaemolyticus, Vibrio azureus, and Providencia rettgeri, while in stool samples they were Enterobacter sichuanensis, Enterobacter hormaechei, Providencia sneebia, and Proteus vulgaris. Providencia stuartii, Proteus mirabilis, Pseudomonas aeruginosa, Escherichia coli, and Klebsiella pneumoniae were common isolates from both seafood and stool samples. This study has shown that not all reported cases of cholera are caused by Vibrio cholerae. Therefore, attention should be paid to other water-borne bacteria in every outbreak, especially in coastal communities.

Identification of β-lactam-resistant coding genes in the treatment plant by activated sludge process

Wastewater treatment plants are the most important release source of antibiotic-resistance genes (ARGs) into the environment. This study aimed to identify β-lactam-resistance genes in Qom province municipal wastewater and also to determine the effect of municipal wastewater treatment plants with different processes on reducing these pollutants. Sampling was performed according to the standard protocol and at constant temperature conditions. Nine genes resistant to the six groups of β-lactam antibiotics were selected, and polymerase chain reaction (PCR) test was performed to identify the presence/absence of ARGs. At the same time, real-time PCR test was also carried out to quantitatively measure three ARGs in wastewater and effluent samples. The obtained results showed that the highest and the lowest removal were related to blanps-1 (90.61%) and blaCTX-M-32 (65.93%), respectively; however, a decreasing trend in these two genes were showed in to blaoxa-1 with an increase in effluent unit (45.12%). The results showed that conventional biological wastewater treatment processes not only have little potential to reduce antibiotic-resistance genes during the treatment process but also these processes sometimes increase resistant genes. The results of the oxa-1 gene in this study are consistent with this trend. In addition, the sludge treatment unit was based on the results of sequencing Salmonella enterica and Klebsiella pneumoniae. Our study suggests that wastewater treatment plants using conventional chlorination do not favor the proliferation of antibiotic resistance bacteria and ARGs during wastewater treatment. Thus, equipping treatment plants with advanced processes will be efficient in reducing bacterial resistance.

Investigation of Antibiotic Resistance Profiles and Carbapenemase Resistance Genes in Acinetobacter Baumannii Strains Isolated From Clinical Samples

Objective: Acinetobacter baumannii is an important pathogen that can develop multiple drug resistance. Here, we aimed to investigate the antibiotic resistance profiles of A. baumannii strains isolated from the various clinics of our hospital and determine the class D beta-lactamase resistance genes causing carbapenem resistance. Methods: Between June-2016 and June-2017, 157 A. baumannii strains isolated from clinical specimens of our hospital were identified with automatic bacterial identification system and antibiograms were determined by the same system. Among the carbapenem resistant strains, blaOXA-51, blaOXA-58, blaOXA-23 and blaOXA-24 genes were also investigated by PCR method. Results: When we analyze the resistance profiles of the strains, we observed that the lowest resistance rate was against colistin with 5 (3.2%) strains. OXA-51 and OXA-23 genes were found positive in all isolates, while OXA-24 was found positive in 16 (32%) strains; OXA-58 was not detected in any of the strains. Conclusion: The most effective antibiotics for carbapenem resistant A. baumannii isolates were colistin, tigecycline and amikacin. Prevelance of OXA-24 enzyme gene was found higher than other similar studies. Monitoring antibiogram profiles and conducting molecular epidemiological studies may help us detect resistant bacteria at the source and reduce the development of resistance.

Clean clothes or dirty clothes? Outbreak investigation of carbapenem-resistant Acinetobacter baumannii related to laundry contamination through multilocus sequence typing (MLST).

To investigate the source in an outbreak of carbapenem-resistant Acinetobacter baumannii (CRA) in a general hospital due to contamination of a laundry evaporative cooler and the laundry environment using multilocus sequence typing (MLST). For CRA culture, clinical samples were collected from infected patients and close contacts, and environmental sampling was performed in patient surroundings and laundry facilities. MLST was used for the molecular typing of representative CRA isolates. Bacterial isolates with identical sequence types were considered epidemiologically linked and attributable to the same source. OXA genes in Acinetobacter baumannii were detected using polymerase chain reaction (PCR). In total, 58 patients were affected in this outbreak. The mean patient age was 75.3, and 50% were female. The most common diagnoses at admission were skin and soft-tissue infection (n = 12, 20.7%) and pneumonia (n = 12, 20.7%). OXA-23 was positive in 64.7% of isolates. A CRA isolate from the evaporative cooler in the laundry was identical to that of 11 patients across 3 wards, belonging to ST345. Isolates from 3 laundry linen racks were identical to those of 7 patients from 3 wards, classified as ST1145. Isolates found on another linen rack and a pajama shelf were identical to isolates from 3 other patients from 2 wards, belonging to ST2207. There was no significant difference between sequence type distributions of clinical and environmental isolates (P = .12), indicating high likelihood of CRA originating from the same source. MLST confirmed that contamination of the laundry evaporative cooler and surrounding environment caused a polyclonal CRA hospital outbreak. Hospital laundry is an important area for infection control and outbreak investigations of CRA.

The molecular analysis of antibiotic resistance and identification of the aerobic bacteria isolated from pleural fluids obtained from patients.

Pleural effusion is a common clinical condition due to various etiological causes. Infectious pleural effusion can be seen in 20-40% of patients. In this study, follow-up of patients admitted to our hospital and diagnosed with pleural effusion are reported. It was aimed to investigate the prevalence of bacteria isolated from patients with pleural effusion and their antibiotic resistance profiles. The pleural fluids obtained from the patients during surgical operations were analyzed microbiologically. Conventional culture, Vitek 2, 16S rRNA, and single Polymerase Chain Reaction (sPCR) were used for microbiological analysis. Twenty-two (12.2%) bacteria were isolated from 180 patients. The most prominent of them were 16 (8.8%) Klebsiella pneumoniae strains. As for the antibiotic sensitivity, gram-negative bacteria showed the highest sensitivity to colistin, while Gram-positive bacteria showed sensitivity to different antibiotics. In 16S rRNA PCR, 22 samples were found to be positive. In the analysis of antibiotic resistance genes, the OXA-48 gene was determined as the highest. In our region, it is essential to perform a microbiological analysis of the sample in patients with pleural effusion. It was thought that revealing both the phenotype and genotype of the antibiotic resistance of the patients was important in terms of treatment. In hospital surveillance, it was considered important to reveal and record the resistance gene profiles of the patients.

Characterization of a genomic Island carrying the tet(X4) gene in porcine Acinetobacter towneri co-harboring plasmid-borne bla NDM-1 and bla OXA-58 genes.

Tigecycline and carbapenems are last-resort antimicrobial agents to treat serious infections caused by multi-drug resistant bacterial pathogens. However, the co-occurrence of tigecycline and carbapenem resistance determinants challenges the clinical efficacy of these antimicrobial agents. In this study, we report the co-existence of tet(X4), blaNDM−1 and blaOXA−58 genes in the porcine Acinetobacter towneri isolate 19110F47. Sequence analysis revealed that tet(X4) gene, along with the florfenicol resistance gene floR, was flanked by three copies of IS91-like elements, which can form three different translocatable units (TUs), and were located in a 41,098-bp multidrug resistance region (MDRR) within a novel 100,354-bp genomic island (GI) region. TUs comprising floR-virD2-ISVsa3, hp-abh-tet(X4)-ISVsa3 and virD2-floR-ISVsa3-hp-abh-tet(X4)-ISVsa3 can be looped out from the chromosomal DNA and facilitate the transfer of the TU-based resistance genes into other plasmidic or chromosomal sites. In addition, the carbapenemase genes blaNDM−1 and blaOXA−58 were found on different non-conjugative multiresistance plasmids in this isolate, with the genetic contexts ISAba125-blaNDM−1-bleMBL-tnpR and ΔISAba3-blaOXA−58-ISAba3, respectively. The simultaneous occurrence of tet(X4), blaNDM−1 and blaOXA−58 in the same porcine A. towneri isolate emphasizes the importance of antimicrobial resistance surveillance in food-producing animals.

Metabolomic profiling of polymyxin-B in combination with meropenem and sulbactam against multi-drug resistant Acinetobacter baumannii.

Empirical therapies using polymyxins combined with other antibiotics are recommended in the treatment of Acinetobacter baumannii infections. In the present study, the synergistic activities of polymyxin-B, meropenem, and sulbactam as combination therapy were investigated using metabolomic analysis. The metabolome of A. baumannii was investigated after treatment with polymyxin-B alone (2 mg/l), meropenem (2 mg/l) alone, combination of polymyxin-B/meropenem at their clinical breakpoints, and triple-antibiotic combination of polymyxin-B/meropenem and 4 mg/l sulbactam. The triple-antibiotic combination significantly changed the metabolite levels involved in cell outer membrane and cell wall biosynthesis, including fatty acid, glycerophospholipid, lipopolysaccharide, peptidoglycan, and nucleotide within 15 min of administration. In contrast, significant changes in metabolome were observed after 1 h in sample treated with either meropenem or polymyxin-B alone. After 1 h of administration, the double and triple combination therapies significantly disrupted nucleotide and amino acid biosynthesis pathways as well as the central carbon metabolism, including pentose phosphate and glycolysis/gluconeogenesis pathways, and tricarboxylic acid cycle. The addition of sulbactam to polymyxin-B and meropenem combination appeared to be an early disruptor of A. baumannii metabolome, which paves the way for further antibiotic penetration into bacteria cells. Combination antibiotics consisting of sulbactam/meropenem/polymyxin-B can effectively confer susceptibility to A. baumannii harboring OXA-23 and other drug resistant genes. Metabolomic profiling reveals underlying mechanisms of synergistic effects of polymyxin-B combined with meropenem and sulbactam against multi-drug resistant A. baumannii.

Evaluation of ceftazidime/avibactam for treatment of carbapenemase-producing carbapenem-resistant Enterobacterales with OXA-48 and/or NDM genes with or without combination therapy.

BackgroundCarbapenem-resistant Enterobacterales (CRE) is an urgent public health threat of significant global concern. Few observational studies have evaluated the clinical outcomes for treatment of CRE harbouring OXA-48 or NDM genes with ceftazidime/avibactam. Previous findings showed lower 30 day mortality with ceftazidime/avibactam ranges between 8.3% and 22%.MethodThis single-centre retrospective cohort study included adult patients aged ≥18 years admitted to King Abdulaziz Medical City (KAMC) who had received ceftazidime/avibactam for at least 72 h for infections caused by CRE with genes encoding for carbapenemase production (CP-CRE).ResultsA total of 211 patients, mostly male (57%), having CP-CRE infections treated with ceftazidime/avibactam were included, with an average age of 62 years. More than 50% of patients were critically ill, for which 46% received invasive ventilation and 36% were on inotropes. The most frequent infectious disease was hospital/ventilator-acquired pneumonia with Klebsiella pneumoniae being the most frequent causative pathogen. The majority of isolates harboured OXA-48 (81%), followed by NDM ± OXA-48 (19%). The overall clinical cure and 30 day mortality was 78% and 21% respectively (stratified per gene: 79% and 21.6% for OXA-48 and 75% and 17.5% for NDM ± OXA-48).ConclusionsThis was the largest study that evaluated clinical outcomes associate with CP-CRE harbouring OXA-48 gene infections treated with ceftazidime/avibactam. Clinical cure and 30 day mortality were consistent with those of previous studies. Findings suggested that combination therapy with ceftazidime/avibactam had no direct impact on clinical outcomes for CP-CRE with OXA-48.

Faecal carriage of multidrug-resistant bacteria and associated risk factors: results from a point prevalence study.

Since 2003, incidences of carbapenemase-producing Gram-negative bacilli (CP-GNB) and vancomycin-resistant Enterococcus faecium (VRE) have steadily increased in France. We therefore conducted a point prevalence study to estimate carriage rates of CP-GNB, VRE and ESBL-producing Enterobacterales (ESBL-PE) and associated risk factors. Between September 2019 and January 2020, all inpatients hospitalized on a given day in 11 teaching hospitals in the Paris urban area were eligible. Patient interviews and rectal swab screening results were recorded by dedicated nurses. The swabs were plated onto selective chromogenic media and processed using the GeneXpert® system. Of 2396 patients, 364 (15.2%) yielded at least one multiresistant bacterial isolate, including 29 CP-GNB carriers (1.2%), 13 VRE carriers (0.5%) and 338 ESBL-PE carriers (14%). In 15 patients (4.4% of ESBL-PE carriers and 36.6% of CP-GNB/VRE carriers), concomitant CP-GNB/VRE and ESBL-PE carriage was observed. In 7/29 CP-GNB and 7/13 VRE carriers, carbapenemase production and vanA in the screening samples was only detected with Xpert® tests. The OXA-48 gene was predominant in 13/34 CP-GNB isolates from 29 carriers. From the 338 ESBL-PE carriers, 372 isolates were recovered, mainly Escherichia coli (61.2%). Among 379 children, 1.1% carried a CP-GNB/VRE strain, and 12.4% carried an ESBL strain. Previous hospitalization outside mainland France, previous antimicrobial treatment and previous ESBL-PE carriage were the main risk factors associated with CP-GNB and/or VRE carriage. The low CP-GNB and VRE prevalence likely reflects the French policy to limit intrahospital spread of CP-GNB and VRE strains.