AmpC Resistance Research Articles

Conventional and unsupervised artificial intelligence analyses identified risk factors for antimicrobial resistance on dairy farms in the province of Québec, Canada

Antimicrobial resistance (AMR) is one of the greatest threats to global health worldwide and is threatening not only humans, but also animal production systems, including dairy farms. The objective of this paper was to describe risks factors associated with AMR on dairy farms in Québec, Canada. This observational cross-sectional study included 101 commercial dairy farms and took place between the springs of 2017 and 2018 for a one-year period. We explored risk factors such as farm practices and producer's knowledge (measured using a questionnaire), antimicrobial use (quantified using veterinary invoices), and the presence of Salmonella Dublin (tested by serology). We evaluated AMR with fecal Escherichia coli retrieved from pre-weaned calves and lactating cows using the following outcomes: the presence of extended-spectrum-β-lactamase/AmpC resistance and the number of resistances to antimicrobial classes. We used logistic regression models to evaluate the association between each risk factor and the 2 outcomes for the 2 types of samples (pre-weaned calves and lactating cows). Furthermore, we explored the relationships between these risk factors utilizing data dimensionality reduction and hierarchical clustering. Outputs of these analyses were used as regressors for AMR in regression models. While the results for univariate analyses were ambiguous, the unsupervised analysis naturally categorized the sample of farms according to their health/treatment status (dimension 1, explaining 12.9% of the variance) and herd size (dimension 2, explaining 7.8%). Three clusters of farms were identified (cluster 1: mainly healthy herds and low ceftiofur users, cluster 2: relatively high ceftiofur users, cluster 3: farms with a higher incidence of diseases and higher antimicrobial treatment rates). Dimension 1 and cluster membership were statistically associated with the presence of extended-spectrum-β-lactamase/AmpC resistance in lactating cows and in pre-weaned calves. Dimension 1 was also statistically associated with the number of resistances in lactating cows and in pre-weaned calves. This study highlights the complexity of analyzing risk factors associated with AMR. Our results suggest that the herd health status and the AMU-related practices used are associated with AMR in dairy farms. However, prospective studies are needed to confirm a causal relation.

Clinical, phenotypic, and genotypic characteristics of ESBL-producing Salmonella enterica bloodstream infections from Qatar

BackgroundResistant Salmonella infections are a major global public health challenge particularly for multidrug-resistant (MDR) isolates manifesting as bloodstream infections (BSIs). ObjectivesTo evaluate clinical, phenotypic, and genotypic characteristics of extended-spectrum beta-lactamase (ESBL) producing Salmonella enterica BSIs from Qatar. MethodsPhenotypic ESBL Salmonella enterica from adult patients presenting with positive BSIs were collected between January 2019 to May 2020. Microbiological identification and characterization were performed using standard methods while genetic characteristics were examined through whole genome sequencing studies. ResultsOf 151 episodes of Salmonella enterica BSI, 15 (10%) phenotypic ESBL isolates were collected. Recent travel was recorded in most cases (80%) with recent exposure to antimicrobials (27%). High-level resistance to quinolines, aminoglycosides, and cephalosporins was recorded (80-100%) while meropenem, tigecycline and colistin demonstrated universal susceptibility. Genomic evaluation demonstrated dominance of serotype Salmonella Typhi sequence type 1 (93%) while antimicrobial resistance genes revealed dominance of aminoglycoside resistance (100%), qnrS1 quinolones resistance (80%), blaCTX-M-15 ESBLs (86.7%), and paucity of AmpC resistance genes (6.7%). ConclusionsInvasive MDR Salmonella enterica is mainly imported, connected to patients from high prevalent regions with recent travel and antimicrobial use caused by specific resistant clones. In suspected cases of multidrug resistance, carbapenem therapy is recommended.

Yokenella regensburgei urinary tract infection in an immunocompetent patient: a case report

Yokenella regensburgei , belonging to the order Enterobacterales , is a rare and emerging human pathogen reported to cause both superficial and invasive infections. The 13 case reports in the literature worldwide highlight blood, bone and wound infections. To our knowledge this is the first case description of Y. regensburgei causing a urinary tract infection in a 69-year-old immunocompetent patient which was isolated in two separate specimens and identified using matrix-assisted laser desorption ionization time-of-flight MS. It was found to be susceptible to most antimicrobials but resistant to penicillin, amoxicillin-clavulanate, cefoxitin and colistin. Inducible chromosomal ampC resistance was demonstrated on disc approximation testing, and blaYOC-1 class C beta-lactamase, beta lactamase superfamily and MBL fold metallo-hydrolase genes were found on whole genome sequencing.

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

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

BlaPSZ-1, a novel AmpC gene identified from a Pantoea isolate.

Pantoea species of the family Erwiniaceae are well-known plant pathogens and animal and human conditional pathogens. Due to the widespread and continuous use of antimicrobials, multidrug-resistant strains continue to emerge, making clinical treatment difficult; therefore, there is an increasing need to clarify the mechanisms of drug resistance. A rabbit anal fecal sample was collected by a swab and the streak plate method was used to isolate single colonies. The standard agar dilution method was used to determine the minimum inhibitory concentrations (MICs) against antimicrobials. The complete genome sequence of the bacterium was obtained using Next-Generation Sequencing platforms. The potential resistance gene was annotated based on the Comprehensive Antibiotic Resistance Database (CARD) and verified by molecular cloning. The β-lactamase PSZ-1 was expressed via the pCold I expression vector and its enzyme kinetic parameters were analyzed. The genetic environment and evolutionary process of the novel resistance gene-related sequences were analyzed by bioinformatic methods. The isolate Pantoea endophytica X85 showed some degree of resistance to penicillins as well as cephalosporins. A novel AmpC resistance gene, designated blaPSZ-1 in this research, was identified to be encoded in the plasmid (pPEX85) of P. endophytica X85. BlaPSZ-1 showed resistance to penicillins and several first-, second-and third-generation cephalosporins as well as aztreonam, but it did not show resistance to the fourth-generation cephalosporins or carbapenems tested. Enzyme kinetic assays revealed that it could hydrolyze amoxicillin, penicillin G, cephalothin, and cefazolin, and its hydrolytic activity could be strongly inhibited by the inhibitor avibactam, which was generally consistent with antimicrobial susceptibility testing results. No hydrolytic activity was observed for third-generation cephalosporins or aztreonam. In this study, a novel AmpC β-lactamase gene, designated blaPSZ-1, was characterized and it was encoded in the plasmid of the bacterium P. endophytica X85. It shows resistance to penicillins and several cephalosporins. The discovery of novel drug resistance mechanisms can help guide the scientific use of drugs in animal husbandry and clinical practice, effectively avoiding the abuse of antimicrobials and thus preventing the further development and spread of bacterial resistance.

Beta-lactam and Fluoroquinolone Resistant Extraintestinal Escherichia coli from Broiler Chickens and Ducks: Public Health Threat

Escherichia coli (E. coli) is a zoonotic bacterial pathogen commonly found in human, birds and animals. The widespread use of broad-spectrum beta-lactams for poultry treatment, has resulted in the production of extended spectrum β-lactamases (ESBLs) in E. coli. In this study, we analyzed the antimicrobial resistance pattern of E. coli isolates from broiler chickens and ducks with clinical colibacillosis, and investigated the presence of beta-lactam (blaTEM, blaSHV, ampC, and blaCTX-M) and fluoroquinolone (qnrS and qepA) resistance genes using PCR. E. coli isolates showed a higher resistance against beta-lactams: penicillin G and ceftriaxone (97.5%), cephalexin (92.5%), ampicillin (77.5%), amoxicillin/clavulanic acid (62.5%), with somewhat lower resistance for nalidixic acid (45%), ciprofloxacin (35%), and norfloxacin (32.5%) from fluoroquinolone group. Isolates showed considerable sensitivity to cefotaxime (65%) and levofloxacin (55%). All tested isolates carried blaTEM, blaSHV and ampC resistance genes, and this was associated with the phenotypic resistance too. Although some isolates showed phenotypic intermediate resistance to fluoroquinolones, or resistance to only one agent from this class, the qnrS (83.3%) has been detected with high frequency. The blaCTX-M (27.87%) and qepA (16.7%) showed the lowest occurrence and had been detected in isolates from broiler chickens only. The occurrence of these resistant E. coli serotypes in broiler chickens and ducks pose a potential threat to poultry industry, and constitutes a public health risk to human consumers, slaughterhouse and poultry packing plant workers due to dissemination of beta-lactam and fluoroquinolone resistant E. coli serotypes.

Antimicrobial resistant bacteria recovered from retail ground meat products in the US include a Raoultella ornithinolytica co-harboring blaKPC-2 and blaNDM-5

Retail beef and pork, including processed products, can serve as vehicles for the zoonotic foodborne transmission of pathogens and antimicrobial resistant bacteria. However, processed and seasoned products like sausages, are not often included in research and surveillance programs. The objective of this study was to investigate retail ground beef and pork, including processed products, for the presence of common foodborne pathogens and antimicrobial resistant bacteria. We purchased 763 packages of fresh and fully cooked retail meat products during 29 visits to 17 grocery stores representing seven major grocery chains located in west and central Ohio. Each package of meat was evaluated for contamination with methicillin-resistant Staphylococcus aureus (MRSA), Salmonella spp., Enterobacteriaceae expressing extended-spectrum cephalosporin resistance, and carbapenemase-producing organisms (CPO). Only 3 of the 144 (2.1%) packages of fully cooked meat products contained any of these organisms, 1 with an extended-spectrum β-lactamase-producing (ESBL) Enterobacteriaceae and 2 with CPO. Among the 619 fresh meat products, we found that 85 (13.7%) packages were contaminated with MRSA, 19 (3.1%) with Salmonella, 136 (22.0%) with Enterobacteriaceae expressing an AmpC (blaCMY) resistance genotype, 25 (4.0%) with Enterobacteriaceae expressing an ESBL (blaCTX-M) resistance genotype, and 31 (5.0%) with CPO, primarily environmental organisms expressing intrinsic carbapenem resistance. However, one CPO, a Raoultella ornithinolytica, isolated from pork sausage co-harbored both blaKPC-2 and blaNDM-5 on IncN and IncX3 plasmids, respectively. Our findings suggest that fresh retail meat, including processed products can be important vehicles for the transmission of foodborne pathogens and antimicrobial resistant bacteria, including those with epidemic carbapenemase-producing genotypes.

Evaluation of Two Phenotypic Methods for the Detection of Plasmid-Mediated AmpC β-Lactamases among Enterobacteriaceae Isolates.

Objectives AmpC β-lactamases are cephalosporinases that confer resistance to cephalothin, cefazolin, cefoxitin, penicillin, and β-lactamase inhibitor-β-lactam combinations. Even though the AmpC resistance is reported, but the accurate occurrence of AmpC β-lactamases in Enterobacteriaceae members is still unknown. Techniques to identify AmpC producers are still evolving but not yet optimized for the clinical laboratory. Here we aimed to compare the test performance of two different phenotypic methods, that is inhibitor-based assay using boronic acid and disk approximation test for AmpC detection in Enterobacteriaceae isolates from a tertiary hospital microbiology laboratory. Materials and Methods The study includes 137 nonrepeat Enterobacteriaceae strains. Bacterial isolates, that yielded a zone diameter of less than 18 mm for cefoxitin by disk diffusion method were considered potential AmpC producers and further confirmed by phenotype methods—inhibitor-based assay using boronic acid and disk approximation test. A multiplex polymerase chain reaction was used to detect the most common plasmid-mediated AmpC genes: ACC, FOX, MOX, DHA, CIT, and EBC. Results Of the 137 clinical isolates, 58 (42.33%) were cefoxitin resistant, while 53.4 and 18.9% of the cefoxitin-resistant isolates were positive by inhibitor-based assay and disk approximation test. Multiplex PCR detected 42 (30.6%) isolates with AmpC genes. Of the 42 isolates, the inhibitor-based assay detected 25 (59.5%) isolates, while the disk approximation test detected nine (21.4%) isolates. Conclusion Our findings suggest that inhibitor-based assay using boronic acid can be used for the detection of the isolates that harbor AmpC β-lactamases. This method is cost-effective, simple to perform, and easy to interpret. Thus AmpC detection as a routine in clinical laboratories can help in appropriate therapeutic intervention and improved infection control.

Curcumin-meropenem synergy in carbapenem resistant Klebsiella pneumoniae curcumin-meropenem synergy.

Background and Objectives:The frequency of multiple resistant bacterial infections, including carbapenems, is increasing worldwide. As the decrease in treatment options causes difficulties in treatment, interest in new antimicrobials is increasing. One of the promising natural ingredients is curcumin. It is known to be effective in bacteria such as Pseudomonas aeruginosa, Escherichia coli, Burkholderia pseudomallei through efflux pump inhibition, toxin inhibition and enzymes. However, because its bioavailability is poor, it seffectiveness occurs in combination with antibiotics. In the study, the interaction of meropenem and curcumin in carbapenemase producing strains of Klebsiella pneumoniae was tested.Materials and Methods:Thirty-nine Klebsiella pneumoniae isolates, resistant to meropenem, were used in this study. From those 15 MBL, 6 KPC, 17 OXA-48 and 1 AmpC resistance pattern were detected by combination disk method. Meropenem and Curcumin MIC values were determined by liquid microdilution. Checkerboard liquid microdilution was used to determine the synergy between meropenem and curcumin.Results:Synergistic effects were observed in 4 isolates producing MBL, 3 isolates producing KPC, 4 isolates producing OXA-48, and 1 isolates producing AmpC (totally 12 isolates) according to the calculated FICI. No antagonistic effects were observed in any isolates.Conclusion:Curcumin was thought to be an alternative antimicrobial in combination therapies that would positively contribute to the treatment of bacterial infection. The effectiveness of this combination should be confirmed by other in vitro and clinical studies.

Evaluation of the VITEK 2 Advanced Expert System performance for predicting resistance mechanisms in Enterobacterales acquired from a hospital-based screening program.

There is limited literature examining the accuracy of the VITEK 2 Advanced Expert System (AES) in characterisation of β-lactamase resistance patterns. We present a prospective single centre study to better ascertain the performance characteristics of this program. The VITEK 2 AES interpretation was compared to established laboratory phenotypic methods. The overall sensitivity for detection of broad-spectrum β-lactamase by the AES was 95%, with a specificity of 78%. One or more discrepancies were noted in 36% of samples, with the majority of these (87/100) due to incorrect 'overcall' of a resistance mechanism. AES characterisation of AmpC resistance mechanisms was excellent. In contrast, the AES had poor specificity in classifying extended spectrum β-lactamases (ESBLs). As a screening aid, the AES can be a valuable tool. However, optimal use requires an adequate working knowledge of resistance mechanisms in order to correctly interpret and accept the result output.

Metallo-\u03b2-lactamase and AmpC genes in Escherichia coli, Klebsiella pneumoniae, and Pseudomonas aeruginosa isolates from abattoir and poultry origin in Nigeria

BackgroundGram-negative bacteria (GNB) including Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae represent the most relevant reservoir of resistance genes such as metallo-β-lactamase (MBL) and AmpC genes that give them the undue advantage to resist antimicrobial onslaught. This study aimed to investigate the occurrence of MBL (blaIMP-1, blaIMP-2, blaVIM-1, blaVIM-2) and AmpC (blaFOX, blaDHA, blaCMY, blaACC) resistance genes in aforementioned GNB collected from abattoir and poultry sources in Nigeria.ResultsIn total, 370 isolates were collected from abattoir tables (n = 130), anal region of cows (n = 120), and the cloacae of poultry birds (n = 120). The test isolates showed high rate of resistance to cephalosporins and carbapenems. The MBLs were phenotypically detected in 22 E. coli, 22 P. aeruginosa, and 18 K. pneumoniae isolates using combined disc test (CDT). However, only 11 E. coli, 24 P. aeruginosa, and 18 Klebsiella pneumoniae isolates were phenotypically confirmed to be AmpC producers using cefoxitin-cloxacillin double disk synergy test (CC-DDST). MBL encoding genes (particularly the blaIMP-1 genes and blaIMP-2 genes) were detected by polymerase chain reaction (PCR) in 12 (54.6%) E. coli, 15 (83.3%) K. pneumoniae, and 16 (72.7%) P. aeruginosa isolates. AmpC genes (particularly the blaCMY genes and blaFOX genes) were found in a total of 5 (29.4%) E. coli isolates, 5 (27.8%) isolates of K. pneumoniae, and 10 (41.7%) isolates of P. aeruginosa.ConclusionsOur study showed the circulation of MBL and AmpC genes in GNB from abattoir and poultry origin in Nigeria. Adoption of regular control policies is necessary to reduce the spread of these species as soon as possible, especially in poultry and slaughterhouses.

Antimicrobial Resistance Profile and Detection of Extended Spectrum and Amp C β-Lactamase Resistance Genes in Escherichia coli Isolated from Diarrheic Children in Lafia, Nasarawa State, Nigeria

Aims: This study evaluated the presence of extended spectrum β-lactamase (ESBL) and AmpC β-lactamase resistance genes in E. coli from stool of diarrheic children in some hospitals in Lafia metropolis, Nigeria.
 Methodology: A total of 70 stool samples of children were obtained from Dalhatu Araf Specialist Hospital, Lafia, M & D Hospital, Olivet Medical Centre and Sandaji Medical Centre, Lafia. Escherichia coli were isolated and identified using standard microbiological methods. Antimicrobial susceptibility of the isolates was tested using Clinical and Laboratory Standards Institute (CLSI) method. The phenotypic detection of ESBL and AmpC β-lactamase production in some antibiotic resistant isolates were carried out using disc method. The molecular detections of ESBL and AmpC resistance genes were carried out using Polymerase Chain reaction (PCR) method.
 Results: Of the 70 samples, the occurrence of E. coli was 100%. The isolates were highly resistant to ampicillin (97.14%), ciprofloxacin (90.00%), sulfamethoxazole/trimethoprim (84.29%), streptomycin (78.57%), amoxicillin/clavulanic acid (70.00%); moderate to gentamicin (38.57%), ceftazidime (37.14%) and cefotaxime (30.00%); and less resistant to cefoxitin (15.71%) and imipenem (8.57%). Twenty-one (30.00%) isolates were jointly resistant to both cefotaxime and ceftazidine. Of this number, 66.67% (14/21) were phenotypically confirmed ESBL producers; and the occurrences of ESBL resistance genes were: 7.14% (SHV), 42.86% (CTX-M) and 50.00% (TEM). Out of 11isolates resistant to cefoxitin, 4(36.36%) were phenotypically confirmed as AmpC β-lactamase producers; and the occurrence of AmpC genes were: 50.00% (CIT), 25.00% (FOX) and 25.00% (MOX).
 Conclusion: The isolates were least resistant to imipenem and cefoxitin and highly resistant to ampicillin, ciprofloxacin and sulfamethoxazole/trimethoprim. TEM and CTX-M ESBL genes were more frequent than SHV. CIT AmpC gene was more frequent than FOX and MOX.

Antibiotic Susceptibility Testing (AST) Reports: A Basis for Environmental/Epidemiological Surveillance and Infection Control Amongst Environmental Vibrio cholerae.

Distribution, investigation, surveillance and control (DISC) of cholera outbreaks in endemic/non-endemic regions has been a concerted approach towards the management of the causal pathogen. Relevant organization, government, health systems and the public have implemented several steps towards controlling the menace, yet pathogen continues to occur with diverse phenotypes/genotypes of high clinical and epidemiological relevance. The study determines antibiotic susceptibility/resistance pattern of Vibrio cholerae isolates retrieved from six domestic water sources between March and August 2018. Serological and molecular typing methods (polymerase chain reaction or PCR) were used to confirm the isolates identity. Antibiotic susceptibility testing was conducted using six commonly employed antibiotics of V. cholerae according to the recommendation of Clinical Laboratory Standard and European Committee for Antimicrobial Susceptibility Testing with other relevant antibiotics of investigative epidemiology and infection control, employing both disc diffusion test and PCR gene detection. Samples presumptive counts ranged between 1.10 to 7.91 log10 CFU/mL. Amongst the 759 presumptive isolates retrieved, sixty-one were confirmed as V. cholerae which were further serogrouped as Non-O1/Non-O139 V. cholerae. Various V. cholerae resistant phenotypes/genoytypes were detected vis: carbapenemase (CR-Vc; 31.1%/5.3%). New Delhi Metallobetalactamase (NDM-1-Vc; 23.0%/42.5%), extended spectrum betalactamase (ESBL-Vc; 42.6%/blaTEM:86,7%), chloramphenicol resistance (62.3%/Flor: 46.2%}, tetracycline resistance (70.5%/46.7%), AmpC resistance (21.0 (34.4%/56.7%)) and various other resistant genotypes/phenotypes. It was observed that more than 50% of the confirmed V. cholerae isolates possess resistance to two or more antibiotic classes/groups with multiple antibiotic resistance index (MARI) ranging from 0.031 to 0.5. This observation provides necessary information and updates for surveillance, planning and implementation of control strategies for cholera. It would also encourage decision making, formulation of policy by the government and cholera control authorities.

Characterization of Beta-lactamases among MDR gram negative bacilli from a tertiary care hospital in central Kerala

Infections and mortality associated with multidrug- resistant Gram-negative bacilli (MDR GNB) have dramatically increased in all parts of the world. The present study was conducted to screen the burden of Gram negative multidrug resistance in our tertiary care hospital and to pave a stepping stone for the infection control team to tackle the problem effectively. A total of 300 non-repetitive MDR GNB isolates from the specimens received in our diagnostic Microbiology lab oratory for a period of one year were screened phenotypically for Extended spectrum β -lactamases (ESBL), AmpC betalactamases and Carbapenemases. Colistin and Tigecycline susceptibility E-test were also performed randomly on some carbapenemase producing organisms. Among the 300 MDR GNB isolates, ESBL and AmpC producers were 56% and 24% respectively. E.coli was the most common organism with ESBL (73%) and AmpC (46%) resistance mechanisms. Carbapenemase producers were 14.6% with predominant isolate being Acinetobacter spp (40%). Among the risk factors, 13% patients had immunosuppressive conditions like diabetes, 31% had insitu urinary catheters, 57.7% and 29.4% had prior antibiotic usage and hospitalisations respectively. Twenty five carbapenemase producing MDR GNB tested for Colistin MIC were in the susceptible range. 71% of Acinetobacter spp tested for Tigecycline showed resistant MIC values. Multidrug resistance has a significant adverse impact on clinical outcomes of patients & leads to higher costs due to consumption of health-care resources. Adherence to simple measures like hand hygiene & careful attention to barrier precautions can help in preventing the spread of these infections to a greater extent.

Perfil de tolerancia al triclosán y detección de los genes MexA, MexC, AcrB y oqxA relacionados con la expresión de bombas de expulsión en aislados clínicos del género Enterobacter aerogenes y Enterobacter cloacae

Introducción: La resistencia a los antimicrobianos y la tolerancia a biocidas está dada por mecanismos comunes, generados por su uso en diferentes ambientes; mecanismos como la expresión de bombas de expulsión presentes en bacterias del género Enterobacter circulantes amenaza la eficacia de los antimicrobianos limitando las opciones de terapia antibiótica. Objetivos: Determinar el perfil de tolerancia al triclosán y detección de genes asociados a bombas de expulsión en aislados clínicos de Enterobacter aerogenes y Enterobacter cloacae. Materiales y Métodos: Se realizó un estudio observacional, descriptivo y de corte transversal, se determinaron perfiles de tolerancia al triclosán por microdilución, de susceptibilidad antimicrobiana, confirmación fenotípica de mecanismos de resistencia, por reacción en cadena de la polimerasa, se identificó la presencia de genes que codifican para bombas de expulsión. Resultados: El 17% correspondió a Enterobacter cloacae y el 6% Enterobacter aerogenes. El 93,7% de los aislados clínicos del género Enterobacter presentó el fenotipo de resistencia BLEE y AmpC. En el 81,3% de los aislamientos se obtuvo la presencia de al menos un gen relacionado con las expresión de bombas de expulsión, siendo frecuentes MexC y AcrB; no identificó presencia del gen oqxA. Conclusiones: La resistencia a diferentes grupos de antibióticos se identifica en especies de Enterobacter circulante, así la presencia de enzimas BLEE y AmpC, la presencia de genes relacionados con bombas de expulsión y la alta tolerancia al triclosán.

Spread of multidrug-resistant IncHI1 plasmids carrying ESBL gene blaCTX-M-1 and metabolism operon of prebiotic oligosaccharides in commensal Escherichia coli from healthy horses, France

The objective of the study was to identify the genetic determinants and characteristics of expanded-spectrum cephalosporin (ESC) resistance in commensal Escherichia coli from healthy horses in France in 2015. Faecal samples from 744 adult horses were screened for ESC-resistant E. coli isolates. The extended-spectrum beta-lactamase (ESBL)/AmpC resistance genes were identified using polymerase chain reaction (PCR) and sequencing. ESC phenotypes were horizontally transferred by conjugation or transformation. Plasmids carrying ESBL/AmpC genes were typed by PCR-based replicon typing, restriction fragment length polymorphism (RFLP), and plasmid multilocus sequence typing (pMLST). The ESC-resistant E. coli isolates were typed by XbaI macrorestriction analysis. Sixteen of 41 stables harboured at least one horse carrying ESC-resistant E. coli. The proportion of individually tested horses carrying ESC-resistant E. coli was 8.5% (28/328). Fifty non-redundant ESC-resistant E. coli isolates showing a great diversity of XbaI macrorestriction profiles belonged mainly to phylogroup B1, and were negative for major E. coli virulence genes, indicating they are commensal isolates. ESBL blaCTX-M genes were dominant (blaCTX-M-1, n=34; blaCTX-M-2, n=8; blaCTX-M-14, n=2) and located on conjugative plasmids belonging to various incompatibility groups (IncHI1, IncI1, IncN, IncY, or non-typeable). Among these, the multidrug-resistant IncHI1-pST9 plasmids were dominant and simultaneously harboured the blaCTX-M-1/2 genes and an operon enabling the metabolism of short-chain fructo-oligosaccharides (scFOS). In conclusion, commensal E. coli of French horses displayed a significant distribution of IncHI1-pST9 plasmids carrying both the blaCTX-M-1/2 gene and the fos metabolism operon. This finding highlights the risk of co-selection of multidrug-resistant IncHI1 plasmids carrying ESBL genes possibly mediated by the use of scFOS as prebiotic in horses.

Molecular characterization of multidrug-resistant Gram-negative pathogens in three tertiary hospitals in Cairo, Egypt

High rates of antimicrobial resistance (AMR) among Gram-negative pathogens (GNP) have been reported in Egypt. Antimicrobial surveillance and identifying the genetic basis of AMR provide important information to optimize patient care. In this study, we aimed to identify the beta-lactam resistance phenotypes and genotypes of multidrug-resistant (MDR) non-repetitive GNP from 3 tertiary hospitals in Egypt. WZe studied 495 non-repetitive MDR Gram-negative isolates from patients with complicated intra-abdominal infections (cIAI), complicated urinary tract infection (cUTI), and lower respiratory tract infection (LRTI), collected as part of the “Study for Monitoring Antimicrobial Resistance Trends” (SMART) conducted in 3 tertiary hospitals in Cairo, Egypt, from 2015 to 2016. Identification and susceptibility testing of GNP to antimicrobials were tested in each hospital laboratory and confirmed in a reference laboratory (International Health Management Associates (IHMA), Inc., Schaumburg, IL, USA). Molecular identification of extended-spectrum beta-lactamases (ESΒLs), AmpC, and carbapenem resistance genes was conducted in IHMA. Among the 495 MDR isolates, Klebsiella pneumoniae (K. pneumoniae) and Escherichia coli (E. coli) were the most common (52.7% and 44.2%). K. pneumoniae was most susceptible to colistin, amikacin, ertapenem, and imipenem (92.7%, 72.7%, 69.3%, and 64%, respectively). E. coli was most susceptible to colistin (100%), amikacin (94.1%), imipenem (90.4%), and ertapenem (83.6%). ESBL was detected in 96.2% and ESBL genotypes included blaCTX-M-15 (70.1%), blaTEM-OSBL (48.5%), blaSHV-OSBL (27.9%), and blaCTX-M-14 (10.7%). AmpC resistance genes were identified in 9.7% of the isolates, dominated by blaCMY-2 (5.7%). Carbapenem resistance genes were detected in 45.3% of the isolates. In K. pneumoniae, blaOXA-48 dominated (40.6%), followed by blaNDM-1 (23.7%) and blaOXA-232 (4.5%). In E. coli, the most frequent genes were blaNDM-5 (9.6%), blaOXA-181 (5.5%), blaOXA-244 (3.7%), and blaNDM-1 (3.7%). blaKPC-2 was identified in 0.4% of isolates. Notably, 32.3% of isolates carried more than one resistance gene. Our findings emphasize the continued need for molecular surveillance of MDR pathogens, implementation of strict infection control measures, and antimicrobial stewardship policies in our hospitals.

Occurrence of FOX AmpC gene among Pseudomonas aeruginosa isolates in abattoir samples from south-eastern Nigeria

In Nigeria, several investigations have been done about the prevalence of the AmpC enzyme in clinical isolates of Gram-negative bacteria; however, little information is available on the occurrence rate of this important enzyme in abattoir specimens that play a major role in the environmental pollution in Nigeria. This study aimed to evaluate the presence of FOX AmpC-producing Pseudomonas aeruginosa isolates from abattoir samples by both phenotypic method and polymerase chain reaction (PCR). In this study, 360 abattoir samples were analyzed for the isolation of P. aeruginosa strains. Antibiogram was carried out using the disk diffusion technique. The production of AmpC enzymes was phenotypically screened and confirmed using the cefoxitin--cloxacillin double-disk synergy test (CC-DDST). Finally, gene responsible for FOX AmpC enzyme production was investigated using PCR. A total of 147 (40.8%) isolates of P. aeruginosa was recovered from the abattoir samples. Ceftazidime and ciprofloxacin with 45.6 and 19% of susceptibility rates were the most and the less effective antibiotics, respectively. A total of 24 (16.3%) P. aeruginosa isolates were confirmed to phenotypically produce AmpC enzyme. However, the PCR result showed that only three (12.5%) of P. aeruginosa isolates harbored the FOX AmpC gene suggesting the attendance of other AmpC resistance genes. This study reported the first occurrence of P. aeruginosa isolates harboring the FOX AmpC gene in abattoir samples from south-eastern Nigeria. This incident requires the adoption of new policies and measures to prevent the further spread of strains carrying the AmpC gene.

Molecular Identification of MefE and AmpC Resistance Genes in ATCC Bacteria

Molecular Identification of <i>MefE </i>and <i>AmpC </i>Resistance Genes in ATCC Bacteria

Clinical outcomes following treatment of Enterobacter species pneumonia with piperacillin/tazobactam compared to cefepime or ertapenem

BackgroundEnterobacter spp. are a common cause of nosocomial pneumonia and treatment can be complicated by AmpC resistance. Carbapenems are the treatment of choice; however, alternatives are needed. Cefepime has been shown to be non-inferior to carbapenems. There are limited data to support the use of piperacillin/tazobactam. The objective of this study was to determine if piperacillin/tazobactam is non-inferior to cefepime or ertapenem for Enterobacter pneumonia treatment. ObjectivesTo compare the rate of clinical cure in patients with Enterobacter pneumonia receiving definitive treatment with piperacillin/tazobactam, cefepime, or ertapenem. Secondary outcomes included hospital mortality, infection-related length of stay, duration of mechanical ventilation, recurrent pneumonia, and resistance. MethodsRetrospective, single-center study. ResultsOf 114 patients included, 59 received definitive treatment with piperacillin/tazobactam and 55 received cefepime or ertapenem. There was no difference in the proportion of patients who achieved clinical cure in the piperacillin/tazobactam group compared to the cefepime or ertapenem group (76.3% vs. 87.3%, P = 0.13). Treatment group was not associated with clinical cure when controlling for confounders in multivariable logistic regression (adjusted odds ratio [OR] 0.59, 95% confidence interval [CI] 0.15-2.37). The rate of recurrent pneumonia was 11.4% in the piperacillin/tazobactam group and 6.7% in the cefepime or ertapenem group (P = 0.48). Other secondary outcomes did not differ between the groups. ConclusionsIn this retrospective study of patients with Enterobacter pneumonia, clinical cure with piperacillin/tazobactam was comparable to that with cefepime or ertapenem; however, a prospective trial with a larger population is needed to determine if definitive treatment with piperacillin/tazobactam is non-inferior to definitive treatment with cefepime or ertapenem.