ESBL Screening Research Articles

Maternal Colonization of Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae in Preterm Rupture of Membranes.

Maternal colonization by extended-spectrum β-lactamase-producing Enterobacteriaceae (ESBL-E) has risen, and the antimicrobial resistance of ESBL-E is significant. We aimed to evaluate the rates of ESBL-E colonization among women with preterm premature rupture of membranes (PPROM) and of maternal-neonatal vertical transmission. We also aimed to compare obstetrical and neonatal complications among ESBL-E positive versus negative maternal colonization in pregnancies complicated by PPROM. This retrospective study included women with PPROM who were admitted from 2018 to 2022 for expectant management and were screened for ESBL-E recto-vaginal colonization on their admission. Obstetrical and neonatal outcomes were compared between positive and negative ESBL-E pregnancies. Neonatal outcomes were compared between positive and negative ESBL-E neonates. Of 118 women with PPROM, 27 (23%) had positive ESBL-E cultures. ESBL-E isolates (cultures from the placenta, cord, amnion, or uterus) were more common in colonized versus non-colonized ESBL-E mothers (55.6% vs. 11.0%, P < 0.001). ESBL-E isolates were more common in neonates of mothers with positive versus negative ESBL-E cultures (33.3% vs. 4.2%, P= 0.017). A higher proportion of neonates of ESBL-E positive than ESBL-E negative mothers needed antibiotic treatment in the neonatal intensive care unit. Neonatal ESBL-E colonization at birth was apredictor of longer stays in the neonatal intensive care unit (P=0.006). In women with PPROM, maternal-ESBL-E colonization was a significant risk factor for neonatal colonization and was associated with neonatal morbidity. The high maternal colonization rate in PPROM raises the need for routine maternal ESBL screening. Future studies should establish the ideal empiric antibiotic regimen in the neonatal intensive care unit for neonates born to ESBL-E colonized mothers.

Risk factors for Beta-lactam and carbapenem-resistant bacteria causing post-burn infections

Background: Burn patients are predisposed to infection because of the loss of skin barrier protection and low immunity. This study aims to evaluate the risk factors of post-burn infections due to beta-lactam and carbapenem-resistant bacteria (CR) isolated from a major burn unit.Methods: The descriptive study was conducted at Jinnah Hospital Lahore’s Burn and Reconstructive Surgery Centre (JB &amp; RSC) and the Institute of Microbiology and Molecular Genetics (MMG), University of the Punjab, Lahore. The clinical specimens of wound swabs, including the age, gender, socio-economic status, residence, occupation, hospital stay, wards, burn degree, total burnt surface area (TBSA%), etiology of burn, and other factors were recorded. Antimicrobial susceptibility testing (AST), ESBL and CR screening (Beta- lactam and carbapenem resistant bacteria) were performed using CLSI guidelines 2017.Results: The most dominant strain was Pseudomonas spp. 26.5% (n=106), followed by 15% (n=60) Klebsiella spp., and 7.5% (n=30) Acinetobacter spp. with ESBL-producing strains. Compared to nosocomial infections, community-acquired infections were observed in 314 (78.5%) patients with more ESBL production. There was a significant relation between burnt people having an age of ≤ 29 years with ESBL (p=0.985) and carbapenemases (p=0.694) infections. None of the other factors, like low socioeconomic status and more hospital stays had a significant correlation (p &gt; 0.05).Concluded: It has been concluded that MDR gram-negative strains play a serious role in burn infections and demand suitable methods to decrease their number. Pseudomonas spp. was the most common post-burn infection causing bacteria. Unfortunately, a high frequency of burn infections was observed in the patients at a young age.Keywords: Post burn infections; Gram negative; MDR; Beta- lactam; Carbapenem resistant

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

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

Colonization by Extended-Spectrum β-Lactamase-Producing Enterobacterales and Bacteremia in Hematopoietic Stem Cell Transplant Recipients.

Assessing the risk of multidrug-resistant colonization and infections is pivotal for optimizing empirical therapy in hematopoietic stem cell transplants (HSCTs). Limited data exist on extended-spectrum β-lactamase-producing Enterobacterales (ESBL-E) colonization in this population. This study aimed to assess whether ESBL-E colonization constitutes a risk factor for ESBL-E bloodstream infection (BSI) and to evaluate ESBL-E colonization in HSCT recipients. A retrospective analysis of ESBL-E colonization and BSI in HSCT patients was conducted from August 2019 to June 2022. Weekly swabs were collected and cultured on chromogenic selective media, with PCR identifying the β-lactamase genes. Pulsed-field gel electrophoresis (PFGE) and whole-genome sequencing (WGS) assessed the colonizing strains' similarities. Of 222 evaluated HSCT patients, 59.45% were colonized by ESBL-E, with 48.4% at admission. The predominant β-lactamase genes were blaTEM (52%) and blaSHV (20%). PFGE analysis did not reveal predominant clusters in 26 E. coli and 15 K. pneumoniae strains. WGS identified ST16 and ST11 as the predominant sequence types among K. pneumoniae. Thirty-three patients developed thirty-five Enterobacterales-BSIs, with nine being third-generation cephalosporin-resistant. No association was found between ESBL-E colonization and ESBL-BSI (p = 0.087). Although the patients presented a high colonization rate of ESBL-E upon admission, no association between colonization and infection were found. Thus, it seems that ESBL screening is not a useful strategy to assess risk factors and guide therapy for ESBL-BSI in HSCT-patients.

Multidrug-Resistant Bacteria in Surgical Intensive Care Units: Antibiotic Susceptibility and β-Lactamase Characterization.

Multidrug-resistant (MDR) bacteria of the utmost importance are extended-spectrum β-lactamase (ESBL) and carbapenemase-producing Enterobacterales (CRE), carbapenem-resistant Acinetobacter baumannii (CRAB), carbapenem-resistant Pseudomonas aeruginosa (CRPA), methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus spp. (VRE). In this study, an evaluation of MDR bacteria in surgical intensive care units in a tertiary referral hospital was conducted. The study aimed to characterize β-lactamases and other resistance traits of Gram-negative bacteria isolated in surgical intensive care units (ICUs). Disk diffusion and the broth dilution method were used for antibiotic susceptibility testing, whereas ESBL screening was performed through a double disk synergy test and an inhibitor-based test with clavulanic acid. A total of 119 MDR bacterial isolates were analysed. ESBL production was observed in half of the Proteus mirabilis, 90% of the Klebsiella pneumoniae and all of the Enterobacter cloacae and Escherichia coli isolates. OXA-48 carbapenemase, carried by the L plasmid, was detected in 34 K. pneumoniae and one E. coli and Enterobacter cloacae complex isolates, whereas NDM occurred sporadically and was identified in three K. pneumoniae isolates. OXA-48 positive isolates coharboured ESBLs belonging to the CTX-M family in all but one isolate. OXA-23 carbapenemase was confirmed in all A. baumannii isolates. The findings of this study provide valuable insight of resistance determinants of Enterobacterales and A. baumannii which will enhance surveillance and intervention strategies that are necessary to curb the ever-growing carbapenem resistance rates.

The Prevalence of Virulence Factors among ESBLs-Producing Klebsiella pneumoniae Isolated from Khorramabad Hospitals, Iran.

Klebsiella pneumoniae is one of the opportunistic bacteria that cause hospital infections. Various virulence factors are involved in its pathogenesis. The purpose of our study was to investigate the prevalence of virulence factor genes entB, mrkD, magA, kfu, iutA, rmpA, ybtS, and k2 among ESBLs-producing Klebsiella pneumoniae isolated from clinical samples of Khorramabad hospitals in Iran. In this descriptive, analytical study, the extended-spectrum β-lactamases (ESBLs) screening phenotypic test was performed by disc diffusion method, and virulence factor genes were detected by PCR method for Klebsiella pneumoniae isolates. Finally, the obtained data were statistically analyzed by SPSS software (version 21) using the chi-squared test. One hundred and ten K. pneumoniae strains were isolated from urine 69 (62.7%), sputum 16 (14.5%), blood 10 (9.1%), wound 9 (8.2%), tissue 3 (2.7%), body fluids 2 (1.8%), and catheter 1(0.9%). Based on an ESBL screening phenotypic test, 57 (51.8%) K. pneumoniae isolates were ESBLs-producing. Among 57 ESBLs-positive K. pneumoniae, the occurrence of mrkD, entB, ybtS, iutA, kfu, k2, and rmpA genes were 88 (80%), 87 (79.1%), 65 (59.1%), 42 (38.2%), 19 (17.3%), 5 (4.5%), 3 (2.7%), and 2 (1.8%), respectively. The findings of this study revealed, there is no significant association between the occurrence of virulence genes or source of samples with production of ESBLs among K. pneumoniae isolates.

Associations between β-Lactamase Types of Acinetobacter baumannii and Antimicrobial Resistance.

Background and objective: Acinetobacter baumannii (A. baumannii) is an important nosocomial pathogen that not only possesses intrinsic resistance to many classes of antibiotics, but is also capable of rapidly developing antimicrobial resistance during treatment. The aim of this study was to determine the characteristics of resistance of A. baumannii strains to β-lactams and other tested antibiotics, to evaluate the associations between the phenotypes of resistance to β-lactams and other tested antibiotics, and to evaluate the changes in antibiotic resistance of A. baumannii strains over 5 years by comparing the periods of 2016-2017 and 2020-2021. Materials and methods: A total of 233 A. baumannii strains were isolated from different clinical specimens of patients treated at the Hospital of Lithuanian University of Health Sciences in 2016-2017 (n = 130) and 2021-2022 (n = 103). All clinical cultures positive for A. baumannii were analyzed. The type of β-lactamase was detected by phenotypic methods using ESBL plus AmpC screen disk tests and the combination meropenem disk test. Results: In both periods, all A. baumannii strains were resistant to ciprofloxacin; resistance to carbapenems, piperacillin/tazobactam, gentamicin, and tobramycin was noted in more than 80% of strains. A comparison of two periods showed that the percentages of A. baumannii strains producing two or three types of β-lactamases were significantly greater in 2021-2022 than in 2016-2017 (94.2% and 5.8% vs. 17.7% and 2.3%, respectively, p < 0.001). Isolates producing two or three types of β-lactamases were more often resistant to tigecycline, tetracycline, and doxycycline than strains producing one type of β-lactamase (p < 0.001). Conclusions: The frequency of isolation of A. baumannii strains producing two different types of β-lactamases (AmpC plus KPC, AmpC plus ESBL, or ESBL plus KPC) or three types of β-lactamases (AmpC, KPC, and ESBL) and the resistance rates to ampicillin/sulbactam, tigecycline, tetracycline, and doxycycline were significantly greater in 2020-2021 as compared with 2016-2017. The production of two or three types of β-lactamases by A. baumannii strains was associated with higher resistance rates to tetracyclines.

Comparison of Antibiotic-Resistant Pattern of Extended Spectrum Beta-Lactamase and Carbapenem-Resistant Escherichia Coli Isolates from Clinical and Non-Clinical Sources

The increasing rate of antibiotic resistance among E. coli especially those mediated by extended spectrum beta-lactamases and carbapenem-resistant (CR) presents a major threat to public health and healthcare delivery globally. The aim of this research was to compare the antibiotic resistant pattern of extended spectrum beta-lactamase and carbapenem-resistant Escherichia coli isolates from clinical and non-clinical sources. A total of two hundred and fifteen (215) clinical and non-clinical samples were collected for the study. The collected samples were analysis using Standard Microbiological protocol for isolation and identification. Phenotypic detection of ESBL Production and carbapenem resistant was performed using Double Disk Synergy Test (DDST) and Modified Hodge Test (MHT) respectively. Antibiogram studies of ESBL producing and Carbapenem-resistant E. coli was determined using the Kirby–Bauer disk diffusion method. The result of isolation and characterization revealed higher occurrence rate of clinical isolates of E. coli 66.1 % over non-clinical sample 54.0%. Phenotypic ESBL screening of isolated E. coli revealed overall detection rate of 35(30.4 %) and 13(13.0 %) in clinical and non-clinical source respectively while overall detection rate of carbapenem resistant ESBL producers accounted 9.6 % and 8.0 % in clinical and non-clinical isolates respectively. The isolates exhibited high percentage of resistance to nitrofurantion 100 %, cefepime 100 % colistin 66.7 %, amikacin 50.0 % and also exhibit MDR with MARI value of ≥ 0.5. Comparison of antibiotic resistant pattern of carbapenem-resistant extended spectrum beta-lactamase producing E. coli isolates from clinical and non-clinical sources showed no statistical significant difference P-value &lt; 0.05 but were 100 % susceptible to ciprofloxacin and gentamicin. The pattern of similar MDR portrayed by clinical and non-clinical isolate should not be overlooked due to its hyper-motility and virulence domination leading to possible AMR transfer to other bacterial organisms and also, appropriate measures such as pretreatment of animal excrement before being used as fertilizers and the quality of irrigation water, hospital effluent discharge need to be taken into consideration to prevent the spread of resistant determinant.&#x0D; Keywords: Escherichia coli, Extended Spectrum Beta-Lactamase, Carbapenem-Resistant, clinical, non-clinical

Pathogenic Bacteria and Their Antibiotic Resistance Patterns in Milk, Yoghurt and Milk Contact Surfaces in Debre Berhan Town, Ethiopia.

Bacterial contamination of milk is a primary culprit for causing foodborne illnesses, presenting a significant health hazard for millions of individuals around the globe. The level and variety of microorganisms present in raw milk determine its degree of contamination and the potential health risks it poses. A cross-sectional survey was conducted from February to August. A questionnaire was used to collect data on socio-demographic characteristics and hygiene practices from milk distributors and traders. Raw milk, yoghurt, swabs from milk containers and drinking cups were collected and processed for bacterial isolation and identification, antibiotic susceptibility testing, MDR screening and confirmation, ESBL screening and confirmation. Finally, all data were pooled and analyzed using SPSS software version 25. A total of 120 samples of fresh milk, yogurt and cotton swabs from milk containers and cups were collected. A total of 80 bacterial isolates were isolated from 120 samples. Among the bacteria isolated, S. aureus 17 (21.3%), E. coli 17 (21.3%), S. epidermidis 14 (17.5%), Klebsiella spp. 9 (11.3%) and Salmonella spp. 7 (8.8%) were detected most often. High rate of contamination was observed in fresh milk 23 (28.8%) and yogurt 23 (28.8%). All isolates were resistant to at least one antibiotic tested. Comparatively, high rates of resistance were observed in all isolates to the most commonly prescribed antibiotics in Ethiopia. However, lower rates of resistance have been observed for recently introduced antibiotics in Ethiopia. Of the isolates, 20 (25.0%) were resistant to eight or more antibiotics. While 16 (20.0%), 12 (15.0%), 9 (11.3%) isolates were resistant to two, three and five antibiotics, respectively. Of the bacteria isolated, 52/80 (65.0%) were MDR, 25/49 (51.0%) were screened for ESBL production, and 20/49 (40.8%) isolates were confirmed as ESBL producer. This study showed a high rate ofbacterial isolates along with MDR and ESBL-producing strains in raw milk, yoghurt, milk container swabs and drinking cup swab samples, associated with poor hygiene and sanitation practices.

High Levels of Multidrug-Resistant and Beta-Lactamase-Producing Bacteria in Meat and Meat Contact Surfaces, Debre Berhan Town, Ethiopia.

Over the years, microbial contamination caused by foodborne bacteria has led to a significant number of food recalls, particularly for meat items that have been related to outbreaks of deadly diseases. Animals often carry Salmonella and Escherichia coli bacteria in their intestines, and these bacteria contaminate raw meat during slaughter. In addition, pathogens such as Staphylococcus aureus can contaminate meat processing equipment and utensils and spread to raw meat. A cross-sectional study was undertaken between 30 February and 15 March 2022. Sanitary conditions of abattoir and butchers and food handlers' hygienic practices were assessed using a structured questionnaire. An equal number of meat, swabs from carcasses, knife, weighing balance and cutting board samples (24 each, 120 total) were collected from abattoir and butcher shops. The collected samples were processed for bacterial isolation, antimicrobial susceptibility testing, MDR screening and confirmation, and ESBL screening and confirmation. Finally, SPSS software version 25 was used to compile and analyze the data. Descriptive data from surveys and laboratory procedures were cross-tabulated and summarized using statistical tables and figure. A total of 76 bacteria were isolated from 120 samples. Of all bacteria isolated, S. aureus 16 (21.1%). E. coli 13 (17.1%), and S. epidermidis 12 (15.8%) were the most prevalent. The rate of bacterial contamination was high in meat 18 (23.7%), carcasses 15 (19.7%) and weighing balance 15 (19.7%), respectively. Among the isolates, 18 (23.7%) were resistant for eight and more antibiotics. While, 17 (22.4%), 7(9.2%) and 4(5.3%) of the isolates were resistant for two and three, four and five, and six and seven antibiotics, respectively. Of bacteria isolated, 51/76 (67.1%) were MDR, 23/48 (47.9%) were screened for ESBL production and 13/48 (27.1%) isolates were confirmed as ESBL producer. Multidrug-resistant bacterial contamination was common in meat and meat contact surfaces, which was exacerbated by inadequate sanitation and hygiene practices.

Occurrence of extended-spectrum β-lactamase-producing Enterobacteriaceae in raw milk from cows with subclinical mastitis in northeast Brazil.

Extended-spectrum β-lactamase (ESBL)-producing Gram negative bacteria are becoming increasingly important in veterinary and human medicine because they can hydrolyze the third generation β-lactams, penicillins, and monobactams. The aim of this study was to identify ESBL-producing Enterobacteriaceae in raw cow milk samples from northeast Brazil. Twenty-six bacterial isolates belonging to the Enterobacteriaceae family were obtained from milk samples from 257 cows with subclinical mastitis. Using microbiological tests, 53.85% (14/26) were identified as Escherichia coli, 15.38% (4/26) as Proteus mirabilis, 26.92% (7/26) as Klebsiella spp., and 3.85% (1/26) as Citrobacter spp. Of all the isolates, 61.54% (16/26) were positive in the ESBL screening test, of which 12.5% (2/16) were positive in the double-disc synergy test using three types of cephalosporins and amoxicillin/clavulanic acid. The two isolates were identified as Klebsiella spp. Among all the isolates, 53.85% (14/26) were positive for one or both ESBL-encoding genes, blaSHV and blaTEM; among these, 71.43% (10/14) were identified as E. coli. This study demonstrates that ESBL-producing bacteria can be found in raw cow milk from northeast Brazil. Cows with subclinical mastitis should be recognized as reservoirs of these strains, which can propagate to humans.

Smartphone-operated affordable PCR thermal cycler for the detection of antimicrobial resistant bacterial genes.

Antimicrobial resistance (AMR) is a global public health threat. Surveillance of AMR requires affordable, rapid, and user-friendly diagnostic methods. Our aim was to develop a low-cost thermocycler to perform polymerase chain reaction (PCR). We developed a smartphone-operated PCR thermal cycler using locally available recycled materials. The thermal cycler was used for the amplification for three bacterial genes-bla-TEM, bla-CTXM and 16s rRNA in human urine samples. The performance of custom-built thermal cycler was compared with commercial thermal cycler. The thermal cycler was portable (<1kg weight), required 12 V power supply, 25 μL of solution, and cost only USD50.0. Temperature and time conditions were instructed using a custom-built smartphone application. The ramping rate of was 0.23°C for heating and 0.43°C for cooling. The reported temperatures were within ± 0.5°C of set temperature. The human urine samples were highly resistance and multi-resistant. Nearly 46% (n = 54) E. coli isolates were positive in ESBL screening test. The custom-built thermocycler was able to accurately predict the presence of bla-TEM, bla-CTXM genes, and 16s rRNA (n = 6). We developed and demonstrated a portable, low-cost, easy-to-use, and smartphone-operated PCR thermal cycler. Since it is portable, it can be used in remote location and field settings, including places without stable power supply. The use of the thermal cycler system can be extended, beyond the detection of AMR genes, e.g., in clinical diagnosis, genetics, forensic analysis, and environmental protection.

Detection of extended spectrum beta-lactamase producing organisms and its antibiotic resistance pattern among Enterobacteriaceae isolates in a teaching hospital of Eastern India

Introduction: ESBL-producing Enterobacteriaceae have been responsible for innumerable outbreaks of infection throughout the world and are becoming a real challenge to control the spread of infection. Aim To determine the prevalence of ESBL producing organisms among Enterobacteriaceae isolated from different clinical samples and to detect their antibiotic susceptibility patterns. Material and Methods The study was conducted in the Department of Microbiology, College of Medicine and JNM hospital. A total of 241 samples showing pure growth of Enterobacteriaceae were included in the study. Identification followed by antibiotic susceptibility tests were performed according to CLSI guidelines. Organisms showing reduced susceptibility to third generation cephalosporins were further tested by double disc synergy test to detect ESBL production and those ESBL screening positive isolates were further tested by Phenotypic confirmatory test according to CLSI 2019 guidelines. Results: Prevalence of ESBL producing Enterobacteriaceae was found to be 31.04% with the highest rate among E.coli (34.5%) followed by K. pneumoniae (27.6%). and P. mirabilis (24.2%) .All the ESBL isolates were sensitive to meropenem and imipenem. Nitrofurantoin (84.2%) was found to be most effective drug for ESBL isolates from urine samples. Among β-Lactam/ β-Lactam inhibitor drugs, amoxicillin/clavulanic acid was found to be less sensitive compared to cefoperazone/sulbactam (67%) . Most of the ESBL isolates were found to be multidrug resistant. Conclusion: Screening of all the gram-negative organisms for ESBL detection and judicial use of antibiotics as per sensitivity report is utmost important to prevent the spread of resistance strains in the community as well as in hospital. Formulation of infection control committee in every hospital may help to combat the present situation.

Five-Hour Detection of Intestinal Colonization with Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae Using the β-Lacta Phenotypic Test: the BLESSED Study.

Extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae (ESBL-PE) intestinal colonization is of particular concern as it negatively impacts morbidity and is the main source of external cross-contamination in hospitalized patients. Contact isolation strategies may be caught out due to the turnaround time needed by laboratories to report intestinal colonization, during which patients may be inappropriately isolated or not isolated. Here, we developed a protocol combining enrichment by a rapid selective subculture of rectal swab medium and realization of a β-Lacta test on the obtained bacterial pellet (named the BLESSED protocol). The performances of this protocol were validated in vitro on 12 ESBL-PE strains spiked into calibrated sample suspensions and confirmed in clinical settings using 155 rectal swabs, of which 23 (reference method) and 31 (postenrichment broth culture) came from ESBL-PE carriers. In vitro, the protocol detected, with 100% sensitivity, the presence of the 12 ESBL-PE strains from 104 CFU/mL. In the clinical validation cohort, 22 out of the 23 (reference method) and 28 out of the 31 (postenrichment broth culture) ESBL-PE-positive rectal samples were accurately detected. The diagnostic performances for ESBL-PE detection, considering all ESBL-PE carriers, were 90% sensitivity, 98% specificity, an 87% positive predictive value, and a 98% negative predictive value. Our protocol is a rapid and low-cost method that can detect intestinal colonization with ESBL-PE in less than 5 h more accurately than the reference method, opening the field for further studies assessing a rapid and targeted isolation strategy applied only to patients with a positive BLESSED protocol result. IMPORTANCE To both improve the efficiency of contact isolation among ESBL-PE carriers and avoid the unnecessary isolation of noncolonized patients, we should reduce the turnaround time of ESBL screening in laboratories and improve the sensitivity of diagnostic methods. The development of rapid and low-cost methods that satisfy these two goals is a promising approach. In this study, we developed such a technique and report its good diagnostic performance, opening the door for further studies assessing a rapid and targeted isolation strategy applied in a few hours only for patients truly colonized with ESBL-producing bacteria.

Comparative study of C-reactive protein and complete blood count in cancer and non-cancer patients followed by antibiogram analysis of isolated bacterial pathogens

Cancer is a group of diseases involving abnormal cell growth with the potential to invade or spread to other parts of the body. This study focuses on the analysis of blood cell and their proteins followed by antibiotic susceptibility and ESBL detection in Cancer and non-Cancer patients. In this study, a total 200 blood and urine samples were screened out for bacteria especially ESBL producing bacteria. Out of the tested samples, different bacterial pathogens were identified and among the isolates E coli were (13.33%), S. aureus (11.66%), P. aeruginosa (11.66%), salmonella (10%), bacillus spps (9.16%), Enterobacter spps (8.33%), Mycobacterium Spps (7.5%), S. Pyogene (7.5%), H. pylori (6.66%), Klebsiella spp. (5.83%), S. epidermidis (4.16 %) and Shigella was (4.16%). The ESBL producing bacteria among the isolated 120 bacterial species were only 14 bacterial isolates are ESBL producers which are E. coli (06), P. aeruginosa (04). Enterobacter spp (04), Klebsiella spp (03) and Shigella were only (01). The 100 blood samples are also analysis through CRP test in which 50 blood samples taken from cancer patients and 50 from non-cancer patients. In total 50 cancer sample CRP analysed that 21 patents have the Lowest (0.92 mg/L), 08 was (0.92-1.93 mg/L) 11 was (1.94-3.69 mg/L) and10 patients were on the Highest (&gt;3.69 mg/L) risk. In 50 non-cancer blood sample the CRP level are 09 individual were (0.92 mg/L), 11 individual (0.92-1.93 mg/L), 25 individual (1.94-3.69 mg/L) and 03 patients were Highest (&gt;3.69 mg/L) risk. Maximum resistivity (91%) was showed by penillion (P) and maximum sensitivity (78%) was showed by rifampicin (RD) against isolated bacterial pathogens. It is recommended that Continuous ESBL screening and supervision are necessary at hospital settings to observe and develop approaches for observing and controlling the spread of ESBL generating bacteria.

413. Cost effectiveness of rectal screening for ESBL producing organism in preventing urosepsis following transrectal prostate biopsy

Abstract Background Urosepsis following transrectal ultrasound guided prostate biopsy (TRUS PB) occurs in 0.3-3.1% of patients. Our center gives ciprofloxacin and ceftriaxone prophylaxis. With rise in urinary pathogens resistant to fluoroquinolones or producing extended spectrum beta lactam (ESBL+), prophylactic regimen might need changing. Studies have highlighted cost effectiveness of rectal screening for ESBL+ organism prior to TRUS PB to determine target antibiotic. We compare our center’s incidence of urosepsis admissions following TRUS PB to the expected rate and calculate cost-effectiveness of pre-procedure screening to prevent urosepsis admissions. Methods All patients with CPT billing code 55700 (biopsy, prostate; needle or punch, single or multiple, any approach) from January 2020 through January 2021 were identified retrospectively. Charts were queried for emergency department or urgent care visits and hospital admissions within two weeks of TRUS PB and admissions reviewed for evidence of urosepsis and ESBL+ cultures. Hospitalization cost was compared to the extrapolated cost of doing pre-procedural rectal swabs. Cost of rectal swab and culture was estimated at $39.02 per swab, and identification and susceptibility at $67 per isolate. Results 33 of 1593 patients presented to our institution within 2 weeks of TRUS PB over a 13 month period. 3 were admitted for urosepsis post TRUS PB (2 with ESBL+ infection, 1 with no growth on culture), 12 had ED visits relating to their TRUS PB without urosepsis, (2 due to UTI with other organisms) and 18 visits were unrelated to TRUS PB. Our urosepsis admission rate was 0.19% (for ESBL+ 0.13%). Cost of the 3 hospitalizations was $37,910.92 ($27,063.76 for 2 ESBL+ infections). Rectal swabs would in theory have prevented 2 admissions. Cost of swab and culture would be $62,158.86. 50% of the swabs would need further testing to identify ESBL+ organism, for an additional cost of $53,365.50. Conclusion Given lower than expected urosepsis admission rates at our institution after TRUS PB, pre-procedure rectal swabs would not be cost effective, especially since some admissions may be unavoidable. The cost analysis is an underestimate as a visit some days prior to procedure would be scheduled and the staff resource utilization to collect the swab is not accounted for. Disclosures All Authors: No reported disclosures.

Prevalence of Extended-Spectrum of Beta-Lactamase-producing class a genes in Escherichia coli and Klebsiella pneumoniae isolated from patients urine samples in United Arab Emirates

The current study aimed to determine the prevalence of class A genes in the Extended Spectrum of β-Lactamase producing E. coli and K. pneumoniae isolated from Patients Urine Samples in United Arab Emirates. A total of 3457 urine samples were collected from October 2018 to January 2019. ESBL screening and confirmatory test were done using MicroScan WalkAway automated bacterial identification system. The ESBL class A gene primers were used to detect the blaCTX-M, blaSHV &amp; blaTEM. Out of the collected urine samples, 474 were reported positive for urinary tract infection. A total of 130 samples were identified as Gram negative ESBL producing species, Escherichia coli (n=107) and Klebsiella pneumoniae (n=23). Amid these 130 ESBL producers, the class A ESBL genes blaTEM, blaCTX-M &amp; blaSHV were detected in 78 isolates of which 59 were E. coli and 19 K. pneumoniae. In the identified 59 E. coli isolates, the most predominant gene was blaTEM, followed by blaCTX-M and blaSHV, and these three genes were detected together in 02 E. coli. In 10 E. coli and 11 K. pneumoniae two class A genes were detected in combination of blaCTX-M + blaSHV, blaSHV + blaTEM and blaTEM + blaCTX-M.

Relationship Of Gentamycin Antibiotic Exposure To: Escherichia Coli Bacteria Resistant To Antibiotic Gentamycin And Escerichia Coli ESBL In Vitro

Background: The development of bacteria that have been resistant to antibiotics can complicate the treatment process. Either causes of antibiotic resistance is inappropriate use of antibiotics. Gentamycin is an aminoglycoside-derived antibiotic which its role is very significant for gram-negative bacteria. Repeated use of gentamycin antibiotics can cause changes the effectiveness of gentamycin so that non ESBL-Gentamycin susceptible Escherichia Coli will change into ESBL-Gentamycin resistant Escherichia coli. Objective: This study aims to prove that repeated exposure to Gentamycin in vitro will change non ESBL-Gentamycin susceptible Escherichia Coli into ESBL Gentamycin resistant Escherichia Coli. Methods: This was an experimental study with 30 samples of non ESBL-Gentamycin susceptible Escherichia coli isolates identified from the Phoenix. Non ESBL- Gentamycin susceptible Escherichia coli was tested by giving exposure to Gentamycin for 14 days, then ESBL screening was tested by Cefotaxime exposure to the results of Gentamycin exposure. Result: There were 4 isolates of Escherichia coli which experienced changes in phenotype into Gentamycin resistant Escherichia coli. The rest of it still susceptible to Gentamycin on days 2, 4 and 10. Furthermore, the Escherichia coli isolates were both susceptible to Gentamycin and those that had phenotypic changes become resistant to Gentamycin after exposed to Cefotaxime as an ESBL screening. There are 8 (26.7%) isolates that are still susceptible to Cefotaxime and 18 (60%) isolates that have been transformed into ESBL-Gentamycin susceptible Escherichia coli. Isolates of 4 (13.3%) Gentamycin-resistant Escherichia coli are then exposed to Cefotaxime and obtained all isolates is resistant to Cefotaxime. Conclusion: Repeated exposure of Gentamycin for 14 days in vitro was not significantly related to the phenotypic changes of non ESBL-Gentamycin susceptible Escherichia coli isolates into ESBL-Gentamycin resistant Escherichia coli (P = 0.550, Phi=0.237). Keywords: Escherichia coli, Susceptible, Resistant, Gentamycin, ESBL

Molecular typing of clinical and environmental isolates of Klebsiella pneumoniae producing ESBLs by PFGE.

Objective(s): Klebsiella pneumoniae is the common cause of pneumonia in hospitalized patients, particularly in intensive care units (ICU). The infection can transfer by medical equipment such as mechanical ventilators. This study aimed to investigate the molecular typing of the extended-spectrum beta-lactamase-producing K. pneumoniae isolates recovered from Beheshti Hospital, Kashan, Iran. Materials and Methods: K. pneumoniae isolates producing ESBLs have been collected from the samples obtained from Shahid Beheshti hospital, Kashan, Iran. Antimicrobial susceptibility was determined using the Kirby Bauer disk diffusion method. The presence of ESBLs was evaluated using CLSI for ESBL screening by the double-disk diffusion method. Molecular typing was conducted by pulsed-field gel electrophoresis (PFGE). In total, 89 K. pneumoniae isolates were recovered, of which 47.1% were ESBL producers.Results:Results showed that all of the clinical and environmental isolates were resistant to ceftriaxone, meropenem, cefazolin, cefotaxime, cephalothin, and piperacillin-tazobactam. All isolates were grouped under four clusters (A-D). The major cluster was related to the C cluster with 22 isolates (19 clinical and 3 environmental). Seventy-two percent of isolates were from the ICU ward. There was no correlation between antibiotic resistance patterns and PFGE clusters (P=0.2).Conclusion:We observed a common molecular signature among both clinical and environmental K. pneumoniae isolates, indicating a similar genotype and likely a common origin for ESBL producer isolates found in different hospital wards. Therefore, hospitals need to implement an effective infection control system to decrease the spreading of ESBL strains within the hospitals and subsequently the transmission of the infection to patients.

Antimicrobial resistant Shigella in North India since the turn of the 21st century

PurposeThe ubiquitous presence and rampant spread of antibiotic resistant strains of Shigella spp is a major public health concern. Therefore, monitoring the trends of antimicrobial resistance in them is essential. MethodsA total of 15440 stool samples were inoculated on MacConkey agar, lysine deoxycholate agar and Selenite F enrichment broth from 2001 to 2015.Out of 491 shigellae isolated, 250 isolates were recovered from our culture collection. Antimicrobial susceptibility was performed by Kirby Bauer disc diffusion method, E-test and phenotypic resistance screening for ESBL and AmpC production was performed. For the detection of beta-lactamase genes, PCR for blaTEM, blaSHV, blaOXA, blaCTX-M-15, CMY-2 and mphA PCR in isolates with decreased susceptibility to azithromycin(DSA) was performed. ResultsS. flexneri (n ​= ​173) was most common, followed by S.dysenteriae (n ​= ​24), S.sonnei (n ​= ​23), S.boydii (n ​= ​10) and Non agglutinating Shigella (NAG, n ​= ​20). A see-saw pattern in the prevalence of S. flexneri and S. dysenteriae and rising prevalence of S. sonnei and NAG was seen. Majority (77%) of the isolates had MICs >4 ​mg/L for ciprofloxacin and >50% had high MIC90 (12 ​mg/L) for ceftriaxone and cefepime (8 ​mg/L). Nearly 20% of S.flexneri were resistant to third generation cephalosporin by disc diffusion and 33.7% had MIC ≥1 ​μg/mL. Among the ceftriaxone resistant isolates (n ​= ​29) the blaTEM beta-lactamase resistance gene was seen in all, blaCTX-M-15 in 37%, blaCMY-2 in 45.6% and blaOXA in 52%. The first report of DSA at our institute was in 2001 (n ​= ​1, 2.5%) which increased to 35.1% (n ​= ​40) in 2011–15. The isolates with DSA included S. flexneri (n ​= ​40), S. boydii (n ​= ​4) and S. sonnei (n ​= ​1) and plasmid mediated resistance to azithromycin by mphA gene was detected in 19 out of 40 isolates of S. flexneri. ConclusionGlobal emergence of resistance Shigella is a matter of concern and calls for systematic monitoring and periodic updates of countrywide and local antibiogram.