Antibiotic-resistant Enterobacteriaceae Research Articles

Molecular detection of extended-spectrum β-lactamase-producing Escherichia coli from bat caves on Lombok Island.

The discovery of antibiotic-resistant Enterobacteriaceae bacteria in wild animals is an indication of their potential for wildlife as a reservoir. Bats are natural reservoir hosts and a source of infection for several microorganisms and have the potential to become vectors for the spread of zoonotic diseases. A study was conducted based on these characteristics to identify and detect the blaTEM gene in Eschericia coli isolated from bat excrements in Tanjung Ringgit Cave, East Lombok. Bat fecal samples were firstly inoculated onto eosin methylene blue agar media. Recovered bacterial isolates were further characterized using standard microbiological techniques. Antimicrobial susceptibility testing was done using the Kirby-Bauer disc diffusion method. blaTEM gene detection was carried out using polymerase chain reaction (PCR). Out of the 150 bat fecal samples obtained from Tanjung Ringgit cave, Lombok Island, Indonesia, 56 (37%) were positive for E. coli. Eight (8) out of the 56 E. coli isolates that underwent antimicrobial susceptibility testing using the disc diffusion method were confirmed to be multidrug-resistant as they exhibited resistance to at least three different classes of antibiotics. Out of the eight (8) multidrug resistance E. coli isolates recovered from fecal samples of bats, 2 (two) harbored the blaTEM gene. The discovery of the blaTEM gene in bat fecal samples indicates the potential for wild animals, especially bats, to spread ESBL resistance genes to the environment and to humans.

Resistance of Enterobacteriaceae to Antibiotics in Wastewaters From the Mindoube Municipal Landfill (Libreville, Gabon)

This study aimed to identify antibiotic-resistant Enterobacteriaceae in wastewater discharged into the landfill. 12 samples were collected from the stream point of tanker truck in The MINDOUBÉ municipal landfill to the river were carried out on 29 September 2022. Bacteriological assays performed at the IRET Microbiology Laboratory yielded one bacterial colony per sample once cultured and isolated. Their identification was as follows: Escherichia coli (58.3%), Serratia Ficaria/Pantoea spp (47.1%), Klebsiella oxytoca (33.3%) and Klebsiella pneumoniae (25%). Antibiotic susceptibility tests showed the presence of the following resistances: Ceftazidime and Cefepime (100%), Cefotaxime (54%), Amoxicillin and Clavulanic Acid (75%). Resistance to sulphonamides, quinolones and aminoglycosides was slight. Therefore, these wastewaters are the source of contamination of the neighbouring stream of the MINDOUBE landfill site. The promiscuity of this rubbish dump and the polluted river is thus having a deleterious effect on the health of the people living nearby, to the extent that they use the water from this river to meet their daily household needs. This suggests that the people living in this neighborhood and using the river water are regularly contaminated by multi-resistant bacteria. This could lead to therapeutic failures in managing infectious diseases triggered by these bacteria.

Assessing the additional health burden of antibiotic resistant Enterobacteriaceae in surface waters through an integrated QMRA and DALY approach

Antibiotic resistant Enterobacteriaceae pose a significant threat to public health. However, limited studies have evaluated the health risks associated with exposure to antibiotic-resistant bacteria (ARB), especially in natural environments. While quantitative microbial risk assessment (QMRA) assesses microbial risks in terms of the probability of infection, it does not account for the severity of health outcomes. In this study, a QMRA-DALY model was developed to integrate QMRA with health burden (disability-adjusted life years (DALY)) from infections caused by ARB. The model considers uncertainties in probability of infection and health burden assessment using Monte Carlo simulations. The study collected antimicrobial resistance (AMR) surveillance data from surface waters with different land uses. Results revealed water bodies with agricultural land use to be the main AMR hotspots, with the highest additional health burden observed in infections caused by meropenem-resistant E. coli (∆DALY = 0.0105 DALY/event) compared to antibiotic-susceptible E. coli. The estimated ∆DALY for antibiotic-resistant K. pneumoniae was lower than for antibiotic-resistant E. coli (highest ∆DALY = 0.00048 DALY/event). The study highlights the need for better evaluation of AMR associated health burden, and effective measures to mitigate the risks associated with antibiotic-resistant bacteria in natural environments.

The Prevalence of Multidrug-Resistant Enterobacteriaceae among Neonates in Kuwait.

Increasing numbers of neonates with serious bacterial infections, due to resistant bacteria, are associated with considerable morbidity and mortality rates. The aim of this study was to evaluate the prevalence of drug-resistant Enterobacteriaceae in the neonatal population and their mothers in Farwaniya Hospital in Kuwait and to determine the basis of resistance. Rectal screening swabs were taken from 242 mothers and 242 neonates in labor rooms and wards. Identification and sensitivity testing were performed using the VITEK® 2 system. Each isolate flagged with any resistance was subjected to the E-test susceptibility method. The detection of resistance genes was performed by PCR, and the Sanger sequencing method was used to identify mutations. Among 168 samples tested by the E-test method, no MDR Enterobacteriaceae were detected among the neonates, while 12 (13.6%) isolates from the mothers' samples were MDR. ESBL, aminoglycosides, fluoroquinolones, and folate pathway inhibitor resistance genes were detected, while beta-lactam-beta-lactamase inhibitor combinations, carbapenems, and tigecycline resistance genes were not. Our results showed that the prevalence of antibiotic resistance in Enterobacteriaceae obtained from neonates in Kuwait is low, and this is encouraging. Furthermore, it is possible to conclude that neonates are acquiring resistance mostly from the environment and after birth but not from their mothers.

Exposure to Non-Antimicrobial Drugs and Risk of Infection with Antibiotic-Resistant Enterobacteriaceae.

Antimicrobial resistance (AMR) has consistently been linked to antibiotic use. However, the roles of commonly prescribed non-antimicrobial drugs as drivers of AMR may be under-appreciated. Here, we studied a cohort of patients with community-acquired pyelonephritis and assessed the association of exposure to non-antimicrobial drugs at the time of hospital admission with infection with drug-resistant organisms (DRO). Associations identified on bivariate analyses were tested using a treatment effects estimator that models both outcome and treatment probability. Exposure to proton-pump inhibitors, beta-blockers, and antimetabolites was significantly associated with multiple resistance phenotypes. Clopidogrel, selective serotonin reuptake inhibitors, and anti-Xa agents were associated with single-drug resistance phenotypes. Antibiotic exposure and indwelling urinary catheters were covariates associated with AMR. Exposure to non-antimicrobial drugs significantly increased the probability of AMR in patients with no other risk factors for resistance. Non-antimicrobial drugs may affect the risk of infection with DRO through multiple mechanisms. If corroborated using additional datasets, these findings offer novel directions for predicting and mitigating AMR.

Profiling and antibiotic resistance of some human intestinal isolates

The cases of antibiotic resistance in Enterobacteriaceae are of great global concern. This study aimed at contributing towards the fight against antibiotic resistance and ameliorate the management/treatment of Enterobacteriaceae-linked diseases. Ten (10) rectal swab samples were collected from five (5) male and five (5) females and two isolates were identified from the samples collected. The isolates were identified through colonial, morphological and biochemical tests carried out following standard procedures. Isolates were investigated for their antibiotic resistance profile, Multiple Antibiotic Resistance indices (MARi), pathogenicity and their resistance genes were identified through molecular means using plasmid amplification and primers. Primers used include. ermB, BlaTem, qnrB genes Result obtained showed the isolates to be Escherichia coli and Klebsiella oxytoca. E. coli showed Alpha (α) heamolysis, while Klebsiella oxytoca showed gamma (γ) heamolysis . E. coli was resistant to 75% of the antibiotics used, Klebsiella oxytoca was sensitive to 42% of all the antibiotics. All the test organisms were resistant to all classes of Cephalosporins. The plasmid profiling revealed that all isolates have low molecular weight plasmids. The molecular fingerprinting of the isolates using gene primers viz -a viz ermB, BlaTem, qnrB genes showed E. coli to have resistance genes for macrolides (ermB gene) while none of the isolates had resistance factor against quinolones (qnrB gene). This study showed a high carriage of Enterobacteriaceae having phenotypic resistance with corresponding plasmid-borne resistance genes. The need to understand how bacteria adapt to the antibiotic environment will lead to new therapeutic strategies for antibiotic-resistant infections. Interventions measures to minimize the abundance of antibiotic-resistant commensals and opportunistic pathogens may include faecal microbiota transplantation and the use of live biotherapeutics.

A cross-sectional study of the antibiotic resistant prevalence of ESBL-producing Enterobacteriaceae in Vietnam

Introduction: Antibiotic resistance in Enterobacteriaceae producing extended spectrum beta-lactamase (ESBL) is increasing. Accurate evaluation of antibiotic resistance rates in various categories of bacteria assists medical physicians in recommending suitable indications for their medical problems, improving treatment efficiency, and minimizing dangers for patients. As a result, we undertook this research to assess the prevalence of ESBL-producing Enterobacteriaceae as well as the rate of antibiotic resistance in ESBL-producing Enterobacteriaceae. Methods: A cross-sectional study was conducted on 2716 patients at the An Giang Central General Hospital from June 2020 to June 2021. Data collection was based on interviews and used SPSS 18.0 and GraphPad Prism 9 for data analysis and presentation. Samples included urine, blood, sputum, and pus. Samples were treated with the Phoenix 100 automated machine to separate and identify samples. Results: The highest rate was 64.8% for Escherichia coli, followed by 30.2% for Klebsiella pneumoniae. Proteus mirabilis and Klebsiella oxytoca were found in 4.5% and 0.6% of the samples, respectively. Ampicillin resistance was greatest in E. coli (96.5%), K. pneumonia (92.4%), and K oxytoca (83.3%). The frequency of resistance to the other antibiotics was likewise extremely high, approaching 60%. Tobramycin, Amoxicillin/Clavulanate, Cefoxitin, and Nitrofurantoin were totally resistant to K. oxytoca in the ESBL-producing group. E. coli and K. pneumoniae with ESBL-producing genes also have a high antibiotic resistance rate of more than 50%. Conclusion: E. coli was the most common pathogenic bacteria. Most of the species of bacteria resisted Ampicillin.

Drug Resistance Mechanism of Enterobacteriaceae with Decreased Antibiotic Sensitivity.

To investigate the mechanism of antibiotic resistance in Enterobacteriaceae. Carbapenem Enterobacteriaceae bacteria isolated from a hospital from January 2015 to June 2020 were selected. Drug resistance phenotype test, drug sensitivity test, and conjugation test were used to observe the drug sensitivity results and the titer of acrB antibody. Finally, the data were statistically analyzed. All strains were resistant to ceftazidime, ceftriaxone, ertapenem, and aztreonam. 87.5% of the samples were resistant to piperacillin. Multisite sequence typing showed that 5 carbapenem-resistant Klebsiella pneumoniae belonged to 4 different types. The sequence types of kpn6099 and kpn6617 were the same. In the sensitivity comparison, Escherichia coli J53 was more sensitive to these two antibiotics, minimum inhibitory concentration values were 0.5 and 0.25 μg/ml, respectively. In addition, the sensitivity of E. coli J53 to carbapenems was slightly higher than that of kpn6617. The results showed that the enzyme-linked immunosorbent assay titer of acrB antibody was 1:40,000, and the preparation of acrB antibody was successful. Plasmid-mediated deletion of IMP-1 metallo-β-lactamase binding outer membrane protein is one of the main reasons for the decrease of antibiotic sensitivity.

Antimicrobial Resistance Profile of Enterobacteriaceae and Drinking Water Quality Among Households in Bule Hora Town, South Ethiopia.

BackgroundAntimicrobial resistance bacteria and unsafe water are contributing to the growing burden of diseases. Hence, isolation and antimicrobial susceptibility tests for drinking water samples are currently becoming vital to address the human health risks from drinking unsafe water.ObjectiveThis study aimed to assess the antimicrobial resistance profile of Enterobacteriaceae and quality of drinking water among households in Bule Hora Town, South Ethiopia.Methods and MaterialsA community-based cross-sectional study was used from June to August 2021. A total of 75 water samples were collected and analyzed at the Microbiological Laboratory of Bule Hora University within 6 hours. A membrane filtration method and biochemical tests were used for the isolation and identification of bacteria. An antimicrobial susceptibility test was performed using Kirby–Bauer disc-diffusion method on Mueller–Hinton Agar. Systematic and simple random probability sampling techniques were used to draw representative samples. The data were entered into Epi Data 3.1 and then exported to SPSS version 25 where univariate and bivariate analyses of the data were conducted.ResultsThis study indicated that 59 (78.6%), 30 (40%), and 20 (26.6%) of water samples from the storage of households were contaminated with total coliforms, thermo-tolerant coliforms, and Escherichia coli, respectively. Similarly, Salmonella accounted for 18 (24%), Shigella 12 (16%), and other members of the Enterobacteriaceae such as Proteus, Citrobacter, Klebsiella, Escherichia coli, and Enterobacter species were also isolated. All isolates of Enterobacteriaceae showed resistance to amoxicillin plus clavulanic acid and ampicillin. Besides, Shigella showed 10 (83.3%), 2 (16.6%) and 10 (83.3%) and Salmonella showed 16 (88.9%), 5 (27.8%) and 15 (83.3%) resistant to cotrimoxazole, chloramphenicol and tetracycline, respectively. All the tested drinking water samples had no free chlorine at all at point of use.ConclusionThere was contamination of drinking water with coliform and antibiotic-resistant Enterobacteriaceae in most surveyed households.

Patterns of Antibiotic Resistance in Enterobacteriaceae Isolates from Broiler Chicken in the West Region of Cameroon: A Cross-Sectional Study.

Background The emergence of multidrug-resistant food-borne pathogens of animal origin including Enterobacteriaceae is a growing concern. Identifying and monitoring resistance in isolates from human-related environments are of clinical and epidemiological significance in containing antimicrobial resistance. This study aimed to contribute towards the fight against antibiotic resistance and ameliorate the management/treatment of Enterobacteriaceae-linked diseases in Cameroon. Methods Cloacal swabs from healthy broilers were enriched in buffered-peptone-water and cultured on EMB agar. Antibiotic susceptibility was tested on Mueller-Hinton-Agar by disc diffusion. Plasmid-borne genes for extended-spectrum beta lactamase (ESBL) and resistance to Quinolones (PMQR) and Aminoglycosides were detected by standard endpoint polymerase chain reaction (PCR). Results A total of 394 isolates were identified belonging to 12 Enterobacteriaceae genera, the most prevalent were Escherichia coli (81/394 = 20.56%), Salmonella spp (74/394 = 18.78%), and Klebsiella spp (39/394 = 9.90%) respectively. Overall, 84/394 (21.32%) were ESBL producers, 164/394 (41.62%) were resistant to quinolones, 66/394 (16.75%) resistant to aminoglycosides with 44.0% (173/394) expressing MDR phenotype. Poor hygiene practice (OR 2.55, 95% CI: 1.67, 3.89, p=0.001) and rearing for >45 days, (OR = 7.98, 95% CI: 5.05, 12.6, p=0.001) were associated with increased carriage of MDR. Plasmid-borne resistance genes were detected in 76/84 (90.48%) of ESBL-producing isolates, 151/164 (92.07%) quinolone resistant isolates and 59/66 (89.39%) aminoglycoside resistant isolates with co-occurrence of two or more genes per isolate in 58/84 (69.05%) of ESBLs, 132/164 (80.49%) of quinolone resistant isolates and 28/66 (42.42%) of aminoglycoside resistant isolates. Conclusion This study found high carriage and widespread distribution of Enterobacteriaceae with ESBL and MDR in broiler chicken in the West Region of Cameroon. Most PMQR genes in bacteria were found at levels higher than is seen elsewhere, representing a risk in the wider human community.

Prevalence of antibiotic resistance in Enterobacteriaceae among the patients hospitalized in Al-Zahra hospital, Isfahan, Iran

Introduction: Enterobacteriaceae are a high prevalent gram-negative species worldwide, which play a major role in various infections. Objectives: The present study aims to assess the prevalence of antimicrobial resistance in Enterobacteriaceae at a tertiary hospital in Isfahan, Iran. Materials and Methods: This cross-sectional laboratory-based study was performed during 2020 in Al-Zahra hospital, Isfahan, Iran. The isolates of all of the clinical samples submitted for the bacterial culture containing Enterobacteriaceae strains were checked for antibiotic resistance during March 2018 to March 2019. Results: The results revealed that the specimens obtained from intensive care units (ICUs) exhibited more resistance than the others. Additionally, the highest abundant antibiotic resistance among inpatients was related to ampicillin/sulbactam, and carbapenem resistance was significantly high. Conclusion: The high prevalence of antibiotic resistance indicated the inappropriate antibiotic prescription in this center. The results emphasized the importance of preventive strategies and restrictions in over-the-counter antibiotic consumption.

W078 Clinically relevant antibiotic resistant enterobacteriaceae from hospital wastewater in metro manila, Philippines

W078 Clinically relevant antibiotic resistant enterobacteriaceae from hospital wastewater in metro manila, Philippines

High Carriage of Extended-Spectrum, Beta Lactamase-Producing, and Colistin-Resistant Enterobacteriaceae in Tibetan Outpatients with Diarrhea.

Antibiotic-resistant bacteria (ARB) and antibiotic-resistance genes (ARGs) have been detected in human-impacted habitats, especially in densely populated cities. The Qinghai–Tibet Plateau is located far from the heavily populated regions of China, and Tibetan residents have distinct dietary habits and gut microbes. Antibiotic-resistance monitoring in the Tibetan population is rare. Here, we collected stool samples from Tibetan outpatients with diarrhea. From 59 samples, 48 antibiotic-resistant Enterobacteriaceae isolates were obtained, including 19 extended-spectrum beta lactamase (ESBL)-producing isolates from 16 patients and 29 polymyxin-resistant isolates from 22 patients. Either ESBL or mcr genes were found in 17 Escherichia coli isolates, approximately 58.8% of which were multidrug-resistant, and ten incompatible plasmid types were found. The gene blaCTX-M was a common genotype in the ESBL-producing E. coli isolates. Four E. coli isolates contained mcr-1. The same mcr-1-carrying plasmid was found in distinct E. coli isolates obtained from the same sample, thus confirming horizontal transmission of mcr-1 between bacteria. Genomic clustering of E. coli isolates obtained from Lhasa, with strains from other regions providing evidence of clone spreading. Our results reveal a strong presence of ARB and ARGs in Tibetan outpatients with diarrhea, implying that ARB and ARGs should be monitored in the Tibetan population.

Genetic Characterization of Antibiotic Resistant Enterobacteriaceae Isolates From Bovine Animals and the Environment in Nigeria.

There is a link between antibiotic resistance in humans, livestock and the environment. This study was carried out to characterize antibiotic resistant bovine and environmental Enterobacteriaceae isolates from Edo state, Nigeria. A total of 109 consecutive isolates of Enterobacteriaceae were isolated from March–May 2015 from 150 fecal samples of healthy bovine animals from three farms at slaughter in Edo state Nigeria. Similarly, 43 Enterobacteriaceae isolates were also obtained from a total of 100 environmental samples from different sources. Isolates were recovered and identified from samples using standard microbiological techniques. Recovered isolates were pre-identified by the Microbact Gram-Negative identification system and confirmed with Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry and ribosomal multilocus sequence typing (rMLST). Antibiotic susceptibility testing was carried out by Kirby-Bauer method for 14 antibiotics. Whole genome sequencing (WGS) was carried out for isolate characterization and identification of resistance determinants. Out of 109 animal and 43 environmental Enterobacteriaceae isolates, 18 (17%) and 8 (19%) isolates based on selection criteria showed antibiotic resistance and were further investigated by whole genome sequencing (WGS). Resistance genes were detected in all (100%) of the resistant bovine and environmental Enterobacteriaceae isolates. The resistance determinants included β-lactamase genes, aminoglycoside modifying enzymes, qnr genes, sulfonamide, tetracycline and trimethoprim resistance genes, respectively. Out of the 18 and 8 resistant animal and environmental isolates 3 (17%) and 2 (25%) were multidrug resistant (MDR) and had resistance determinants which included efflux genes, regulatory systems modulating antibiotic efflux and antibiotic target alteration genes. Our study shows the dissemination of antibiotic resistance especially MDR strains among Nigerian bovine and environmental Enterobacteriaceae isolates. The presence of these resistant strains in animals and the environment constitute a serious health concern indicated by the difficult treatment options of the infections caused by these organisms. To the best of our knowledge we report the first detailed genomic characterization of antibiotic resistance in bovine and environmental Enterobacteriaceae isolates for Nigeria.

High level of intrinsic phenotypic antimicrobial resistance in enterobacteria from terrestrial wildlife in Gabonese national parks.

Data on the prevalence of antibiotic resistance in Enterobacteriaceae in African wildlife are still relatively limited. The aim of this study was to estimate the prevalence of phenotypic intrinsic and acquired antimicrobial resistance of enterobacteria from several species of terrestrial wild mammals in national parks of Gabon. Colony culture and isolation were done using MacConkey agar. Isolates were identified using the VITEK 2 and MALDI-TOF methods. Antibiotic susceptibility was analysed and interpreted according to the European Committee on Antimicrobial Susceptibility Testing guidelines. The preliminary test for ESBL-producing Enterobacteriaceae was performed by replicating enterobacterial colonies on MacConkey agar supplemented with 2 mg/L cefotaxime (MCA+CTX). Extended-spectrum beta-lactamase (ESBL) production was confirmed with the double-disc synergy test (DDST). The inhibition zone diameters were read with SirScan. Among the 130 bacterial colonies isolated from 125 fecal samples, 90 enterobacterial isolates were identified. Escherichia coli (61%) was the most prevalent, followed by Enterobacter cloacae (8%), Proteus mirabilis (8%), Klebsiella variicola (7%), Klebsiella aerogenes (7%), Klebsiella oxytoca (4%), Citrobacter freundii (3%), Klebsiella pneumoniae (1%) and Serratia marcescens (1%). Acquired resistance was carried by E. coli (11% of all E. coli isolates) and E. cloacae (3% of all E. cloacae) isolates, while intrinsic resistance was detected in all the other resistant isolates (n = 31); K. variicola, K. oxytoca, K. pneumoniae, E. cloacae, K. aerogenes, S. marcescens and P. mirabilis). Our data show that most strains isolated in protected areas in Gabon are wild type isolates and carry intrinsic resistance rather than acquired resistance.

High Fecal Contamination and High Levels of Antibiotic-Resistant Enterobacteriaceae in Water Consumed in the City of Maputo, Mozambique.

Simple SummaryThe high number of diarrheal disease cases in developing countries is related to sanitation conditions, consumption of untreated water, and poor individual and collective hygiene. In this study, the microbiological quality of water sold and consumed in the city of Maputo, Mozambique, and the antibiotic resistance profile of Enterobacteriaceae isolated from these samples were evaluated. The results showed the occurrence of microorganisms that indicate fecal contamination with enterococci, fecal coliforms, and Escherichia coli above the limit legally allowed for drinking water. The antibiotic resistance profile revealed the existence of antibiotic-resistant bacteria. These results show the need to improve the water supply system in the city of Maputo and to educate the population on hygiene to reduce health risks and promote well-being. In the city of Maputo, Mozambique, food and water are often sold on the streets. Street water is packaged, distributed, and sold not paying attention to good hygienic practices, and its consumption is often associated with the occurrence of diarrheal diseases. Coincidentally, the increase of diarrheal diseases promotes the inappropriate use of antibiotics that might cause the emergence of antibiotic-resistant bacterial strains. In this context, the present study aimed to assess the microbiological quality of water sold on the streets of Maputo, as well as the antibiotic resistance profile of selected Enterobacteriaceae isolates. The 118 water samples analyzed were from street home-bottled water (n = 81), municipal water distribution systems (tap water) (n = 25), and selected supply wells in several neighborhoods (n = 12). The samples were analyzed for total mesophilic microorganisms, fecal enterococci, fecal coliforms, Escherichia coli, and Vibrio spp. The results showed a high level of fecal contamination in all types of water samples. In home-bottled water, fecal coliforms were found in 88% of the samples, and E. coli in 66% of the samples. In tap water, fecal coliforms were found in 64%, and E. coli in 28% of the samples. In water from supply wells, fecal coliforms and E. coli were found in 83% of the samples. From 33 presumptive Vibrio spp. colonies, only three were identified as V. fluvialis. The remaining isolates belonged to Aeromonas spp. (n = 14) and Klebsiella spp. (n = 16). Of 44 selected Enterobacteriaceae isolates from water samples (28 isolates of E. coli and 16 isolates of Klebsiella spp.), 45.5% were not susceptible to the beta-lactams ampicillin and imipenem, 43.2% to amoxicillin, and 31.8% to amoxicillin/clavulanic acid. Regarding non-beta-lactam antibiotics, there was a high percentage of isolates with tolerance to tetracycline (52.3%) and azithromycin (31.8%). In conclusion, water in Maputo represents a risk for human health due to its high fecal contamination. This situation is made more serious by the fact that a relatively high percentage of isolates with multidrug resistance (40%) were found among Enterobacteriaceae. The dissemination of these results can raise awareness of the urgent need to reduce water contamination in Maputo and other cities in Mozambique.

On-Site Pilot Testing of Hospital Wastewater Ozonation to Reduce Pharmaceutical Residues and Antibiotic-Resistant Bacteria.

Hospital sewage constitutes an important point source for antibiotics and antibiotic-resistant bacteria due to the high antibiotic use. Antibiotic resistance can develop and cause problems in sewage systems within hospitals and municipal wastewater treatment plants, thus, interventions to treat hospital sewage on-site are important. Ozonation has proven effective in treating relatively clean wastewater, but the effect on untreated wastewater is unclear. Therefore, we piloted implementation of ozonation to treat wastewater in a tertiary hospital in Uppsala, Sweden. We measured active pharmaceutical ingredients (APIs) using liquid chromatography-mass spectrometry and antibiotic-resistant Enterobacteriaceae using selective culturing pre- and post-ozonation. Comparing low (1 m3/h) and high (2 m3/h) flow, we obtained a ‘dose-dependent’ effect of API reduction (significant reduction of 12/29 APIs using low and 2/29 APIs using high flow, and a mean reduction of antibiotics of 41% using low vs. 6% using high flow, 25% vs. 6% for all APIs). There was no significant difference in the amount of antibiotic-resistant Enterobacteiaceae pre- and post-ozonation. Our results demonstrate that ozonation of untreated wastewater can reduce API content. However, due to the moderate API decrease and numerous practical challenges in the on-site setting, this specific ozonation system is not suitable to implement at full scale in our hospital.

Antibiotic-Resistant Enterobacteriaceae in Wastewater of Abattoirs.

Antibiotic-resistant Enterobacteriaceae are regularly detected in livestock. As pathogens, they cause difficult-to-treat infections and, as commensals, they may serve as a source of resistance genes for other bacteria. Slaughterhouses produce significant amounts of wastewater containing antimicrobial-resistant bacteria (AMRB), which are released into the environment. We analyzed the wastewater from seven slaughterhouses (pig and poultry) for extended-spectrum β-lactamase (ESBL)-carrying and colistin-resistant Enterobacteriaceae. AMRB were regularly detected in pig and poultry slaughterhouse wastewaters monitored here. All 25 ESBL-producing bacterial strains (19 E. coli and six K. pneumoniae) isolated from poultry slaughterhouses were multidrug-resistant. In pig slaughterhouses 64% (12 of 21 E. coli [57%] and all four detected K. pneumoniae [100%]) were multidrug-resistant. Regarding colistin, resistant Enterobacteriaceae were detected in 54% of poultry and 21% of pig water samples. Carbapenem resistance was not detected. Resistant bacteria were found directly during discharge of wastewaters from abattoirs into water bodies highlighting the role of slaughterhouses for environmental surface water contamination.

Molecular detection of Escherichia coli Cause Urinary Tract Infections among Pregnant Women at Thi-Qar province, Iraq

The prevalence of antibiotic resistance in Enterobacteriaceae has increased sharply in recent years. Extendedspectrum ß-lactamase-producing Enterobacteriaceae include Escherichia coli has become especiallycommon. Although traditionally linked to risk factors such as prior hospitalization and antibiotic use, thesebacteria have become increasingly recognized in the community, especially as pathogens in urinary tractinfections (UTIs). In this study urine samples from 150pregnant women clinically diagnosed with UTIswere used for Gram staining, culture, API 20 E and singleplex PCR. Singleplex PCR was performed withprimers targeted to chuA and yjaA genes and anonymous DNA fragment TspE4C2 of E. coli. The positivesingleplex PCR products were identified by presence of 279 bp, 211 bp and 152 bp amplicons of chuA andyjaA genes and anonymous DNA fragment TspE4C2 for of E.coli. Conventional methods of Gram staining,culture and API 20E test showed positive result for E. coli in 35 (40%) out of 150 pregnant clinicallydiagnosed with urinary tract infection. PCR detected 24 (27.5%) out of the 35 (40%) samples that werepositive for E. coli. The majority of UTIs caused by spectrum ß-lactamase-producing Enterobacteriaceaeinclude E. coli was acquired in the community, so rapid, specific and sensitive molecular are urgently neededto better prevalence, prevent and treat these infections in Iraq

Microbiota restoration reduces antibiotic-resistant bacteria gut colonization in patients with recurrent Clostridioides difficile infection from the open-label PUNCH CD study

BackgroundOnce antibiotic-resistant bacteria become established within the gut microbiota, they can cause infections in the host and be transmitted to other people and the environment. Currently, there are no effective modalities for decreasing or preventing colonization by antibiotic-resistant bacteria. Intestinal microbiota restoration can prevent Clostridioides difficile infection (CDI) recurrences. Another potential application of microbiota restoration is suppression of non-C. difficile multidrug-resistant bacteria and overall decrease in the abundance of antibiotic resistance genes (the resistome) within the gut microbiota. This study characterizes the effects of RBX2660, a microbiota-based investigational therapeutic, on the composition and abundance of the gut microbiota and resistome, as well as multidrug-resistant organism carriage, after delivery to patients suffering from recurrent CDI.MethodsAn open-label, multi-center clinical trial in 11 centers in the USA for the safety and efficacy of RBX2660 on recurrent CDI was conducted. Fecal specimens from 29 of these subjects with recurrent CDI who received either one (N = 16) or two doses of RBX2660 (N = 13) were analyzed secondarily. Stool samples were collected prior to and at intervals up to 6 months post-therapy and analyzed in three ways: (1) 16S rRNA gene sequencing for microbiota taxonomic composition, (2) whole metagenome shotgun sequencing for functional pathways and antibiotic resistome content, and (3) selective and differential bacterial culturing followed by isolate genome sequencing to longitudinally track multidrug-resistant organisms.ResultsSuccessful prevention of CDI recurrence with RBX2660 correlated with taxonomic convergence of patient microbiota to the donor microbiota as measured by weighted UniFrac distance. RBX2660 dramatically reduced the abundance of antibiotic-resistant Enterobacteriaceae in the 2 months after administration. Fecal antibiotic resistance gene carriage decreased in direct relationship to the degree to which donor microbiota engrafted.ConclusionsMicrobiota-based therapeutics reduce resistance gene abundance and resistant organisms in the recipient gut microbiome. This approach could potentially reduce the risk of infections caused by resistant organisms within the patient and the transfer of resistance genes or pathogens to others.Trial registrationClinicalTrials.gov, NCT01925417; registered on August 19, 2013.