Detection of 16S rRNA Methylase Genes in Gram-Negative Bacilli Isolated from Hospitals in Changchun, China

Methylation of 16S rRNA is an important mechanism of aminoglycoside resistance among gram-negative pathogens. In this report, 16S rRNA methylase genes were amplified using PCR among gram-negative bacillus isolates from hospitals in the Changchun area of China and 16S rRNA methylase genotypes (armA, rmtB, rmtA, rmtC, rmtD, and npmA) were identified by direct sequencing. Fifty of the isolates (43.1%) harbored 16S rRNA methylase genes. The common 16S rRNA methylase genes were armA and rmtB (12.1% and 31.0%, respectively), whereas the rmtA, rmtC, rmtD, and npmA genes were absent from the sample. It suggests that the predominant 16S rRNA methylase genes among gramnegative bacilli in the Changchun area are armA and rmtB.

Detection of the Production of Klebsiella Pneumoniae Carbapenemase, New Delhi Metallo-Beta-Lactamase and Oxacillinase-48-Type Carbapenemases by Gram-Negative Bacilli in Resource-Limited Setting

Background: The increasing resistance of bacteria to various antibiotics is a worldwide public health issue. Carbapenems that have elicited great hope in treating infections caused by multidrug-resistant germs have seen their efficacy narrowed over time with the emergence of other novel resistance mechanisms, notably the production of Carbapenemases. Methods: A prospective cross-sectional study was conducted from May 2017 to May 2018 in Douala (Cameroon) to detect carbapenemase-producing Gram-negative bacilli. Isolated strains were identified using the Vitek2TM system. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method on agar plates with 20 selected commercially available antibiotic discs. The bacterial strains were tested for the production of three Carbapenemases (OXA-48, NDM, KPC), using an immuno-chromatographic technique, with the “RESIST-3 O.K.N. K-SeT” rapid detection kit. Results: During the study period, 1687 strains of Gram-negative bacilli were isolated in selected laboratories with a total of 200 multi-resistant strains identified (11.9%). Among the multi-resistant strains, E. coli was the species most represented in Enterobacteriaceae (27.5%) followed by K. pneumoniae (15.5%) and the non-fermenting Gram-negative bacilli were predominantly P. aeruginosa (20.5%). These strains mainly came from urine and pus, i.e. 41% and 32% respectively. Thirty-two (16%) strains produced one of the Carbapenemases with a higher frequency at the General Hospital (84%). NDM-type carbapenemase was the most frequently identified (8.5%), OXA-48 type 7.5%, and no KPC production was observed. Among the Enterobacteriaceae 22.9% produced Carbapenemases and only 5.1% of the non-fermenting bacilli produced these enzymes. The isolates strains were completely resistant to all antibiotics except Amikacin and Fosfomycin. The strains producing the NDM-type carbapenemase showed higher rates of resistance to almost all of the antibiotics tested. Conclusion: Multidrug-resistant strains are experiencing an increase in evolution. The apparition of strains producing Carbapenemases prominently, the NDM and OXA-48 favor this increase. The activities of antibiotics with high efficacies on these strains are low.

Metallo-β-lactamase producing nonfermentative gram-negative bacteria:An increasing clinical threat among hospitalized patients

Objective:To detect and evaluate the various methods for metallo-β-lactamases(MBL) production in Pseudomonas aeruginosa(P.aeruginosa) and Acinetobacter species.Methods:A total of 109 P.aeruginosa and 85 Acinetobacter species were screened for imipenem resistance by Kirby- Bauer disc diffusion methods.Detection of MBL production was(lone by imipenem-EDTA combined disc test,double disc synerygy test(DDST) and imipenem-EDTA MBL E test.Results: A total of 63(57.8%) strains of P.aeruginosa and 46(54.1%) strains of Acinetobacter spp.were found to be resistant to imipenem.Of the 63 imipenem resistant P.aeruginosa tested for MBL production.44(69.89;) were found to be positive and among 46 imipenem resistant Acinetobacter. 19(41.3%) were shown to be the MBL producers.Conclusions:Imipenem-EDTA combined disc test and MBL E test are equally effective for MBL detection in both P.aeruginosa and Acinetobacter spp.,but given the cost-constraints,combined disc can be used as a convenient screening method in the clinical microbiology laboratory.

Serratia marcescens and other non-AACEK GNB endocarditis: A case report and review of literature

BACKGROUND Non-Aggregatibacter aphrophilus,Aggregatibacter actinomycetemcomitans,Cardiobacterium hominis,Eikenella corrodens,Kingella spp.(non-AACEK)gramnegative bacilli(GNBs)are an infrequent and challenging cause of endocarditis associated previously with mainly intravenous drug use.Currently,this pathology has increasingly become a healthcare-associated issue.Current guidelines do not clearly define the management of non-AACEK GNB endocarditis due to a lack of prospective trials.We review characteristics,outcomes and treatment of non-AACEK GNB endocarditis,in particular Serratia marcescens endocarditis.CASE SUMMARY We describe the case report of a 46-year-old man who presented to the emergency department with high-grade fever and a purulent exudate on an intracardiac device site.Serratia marcescens mitral valve endocarditis as a consequence of complicated generator pocket infection was diagnosed.The patient was treated with complete device removal and a long course of broadspectrum antibiotics for 6 wk after surgery with intravenous piperacillintazobactam and ciprofloxacin,which was later switched to oral ciprofloxacin and sulfamethoxazole-trimethoprim.The patient had complete resolution of symptoms and inflammatory parameters at the end of the treatment and at follow-up.CONCLUSION Long-term dual-antibiotic therapy containing a beta-lactam is indicated for most non-AACEK GNB endocarditis, whereas valve surgery may not be necessary inall patients.

The Increased Frequency of Carbapenem Resistant Non Fermenting Gram Negative Pathogens as Causes of Health Care Associated Infections in Adult Cancer Patients

Background and Aim: Multi drug resistant Non fermenting gram negative bacilli (NFGNB) have emerged as a major cause of health-care associated infections especially in immunocompromised hosts. The aim of the study was to investigate the prevalence of NFGNB as a cause of health-care associated infections (HAI) in cancer patients and determine their resistance pattern. Patients and Methods: During the study period, 158 NFGNB isolates were collected. Microscan Walk Away 9 was used for identification and testing for the metallo-β-lactamases (MBLs) was done by Imipenem-EDTA combined disk synergy test (CDST-IPM). Results: NFGNB represented 29.0% of infections caused by gram negative organisms. Carbapenem resistance, the multi-drug resistant (MDR) phenotype, and MBL production were documented in 70%, 63%, and 59% of NFGNB isolates, respectively. MDR-NFGNB rates were significantly higher among hospitalized patients, medical department and those with longer duration of hospital stay (p = 0.034, 0.026, 0.019;respectively) than non MDR-NFGNB. Conclusion: A high level of carbapenem and multi-drug resistance were detected among the non-fermenter pathogens isolated from hospitalized cases and were more frequently encountered in high risk adult cancer patients requiring longer duration of hospitalization. The MDR-NFGNB are constituting important causes of health-care associated infections in cancer patients.

Overview of Multidrug-Resistant Pseudomonas aeruginosa and Novel Therapeutic Approaches

Gram-negative bacilli Pseudomonas aeruginosa is an important pathogen in hospitalized patients, contributing to their morbidity and mortality due to its multiple resistance mechanisms. Therefore, as therapeutic options become restricted, the search for new agents is a priority. Latterly an accelerated increase in frequency of multidrug-resistant clinical strains has severely limited the availability of therapeutic options. Several in vitro and in vitro studies evaluating the efficacy of different antimicrobials agents and development of vaccines against P. aeruginosa have been reported as novel approaches, such as inhibition of virulence factor expression or inhibition of their metabolic pathways.