Activity of vancomycin combined with linezolid against clinical vancomycin-resistant Enterococcus strains.

Introduction: Because multi-drug-resistant Gram-positive bacteria have been isolated frequently worldwide and are difficult to treat, alternative treatment choices are required. Combination antibiotherapies have a distinct advantage over monotherapies in terms of their broad spectrum and synergistic effect. In the present study, it was aimed to investigate the in vitro activity of vancomycin combined with linezolid against clinical vancomycin-resistant enterococci (VRE) strains with high-level aminoglycoside resistance. Material and methods: A total of 30 randomly selected clinical VRE strains were studied. Susceptibility to agents tested was investigated using broth microdilution assay. The inoculum of strain was adjusted to approximately 5 × 105 CFU/ml in the wells. The results were interpreted in accordance with Clinical and Laboratory Standards Institute guidelines. In vitro activities of antibiotics in combination were assessed using the broth microcheckerboard technique. The fractional inhibitory concentration indexes (FICIs) were interpreted as follows: synergism, FICI ≤ 0.5; additive/indifference, FICI ≤ 0.5 - ≤ 4; antagonism, FICI > 4. Results: All strains were resistant to vancomycin and susceptible to linezolid. The MIC50,90 and MICrange values of antimicrobials were 512, 512, and 512-1024 µg/ml for vancomycin; 2, 2, and 2-4 µg/ml for linezolid. The rate of synergy was found to be 46.6% (14/30) for linezolid combined with vancomycin. No antagonism was observed. Conclusions: The results of the study suggest that this combination may contribute to the treatment of VRE infections for their synergistic effect and because no antagonism was observed.

Molecular Characterization of High-Level Aminoglycoside Resistance among Enterococcus Species.

Background Enterococci are nosocomial pathogen. They can develop high-level resistance to aminoglycoside by producing aminoglycoside modifying enzymes (AMEs). In enterococci, high level resistance to aminoglycosides is mediated by acquisition of plasmid mediated genes encoding for aminoglycoside modifying enzymes (AMEs). High level gentamicin resistance (MIC ≥ 500µg /mL) is predominantly mediated by aac(6')-Ie-aph(2″)-Ia, encoding the bifunctional aminoglycoside modifying enzyme AAC(6')-APH(2″). This enzyme eliminates the synergistic activity of gentamicin when combined with a cell wall active agent. Other AME genes such as aph(2″)-Ib, aph(2″)-Ic, aph(2″)-Id and ant(4')-1a have also been detected in enterococci. Objective This study was carried out to determine the diverse prevalence of AME and their pattern of occurrence in the clinical isolates of Enterococci . Materials and Methods A total number of 150 clinical isolates were included in this study. Susceptibility to various antibiotics was determined by disc diffusion. Minimum Inhibitory Concentration (MIC) was ascertained by agar dilution method. Polymerase chain reaction was done to screen the following AMEs (aac(6')-Ie-aph(2″)-Ia; aph(2″)-Ib; aph(2″)-Ic; aph(2″)-Id and aph(3')- IIIa genes) . Results 51.3% of the study isolates exhibited high level gentamicin resistance. Polymerase chain reaction revealed that aph(3')-111a is the most prevalent AME, followed by aac(6')-1e-aph(2″)-1a . The combination of both the genes were detected in 44.1% of the study isolates. The rest of the AMEs and their combinations were not encountered in this study. 8.6% of the study isolates did not harbour any AME genes screened for, but was phenotypically resistant to gentamicin. In contrast 31.3% anchored the AME genes but phenotypically appeared susceptible to gentamicin. Conclusion This study indicates the high- level aminoglycoside resistance disseminated among Enterococci in our geographical region. It also emphasizes the detection of AMEs by PCR is mandatory because strains that appear susceptible by disc diffusion and/or MIC method may harbour one or more AMEs genes leading to therapeutic failure.

Characteristics of High-Level Aminoglycoside-Resistant Enterococcus faecalis Isolated from Bulk Tank Milk in Korea.

Enterococci, which are considered environmental mastitis-causing pathogens, have easily acquired aminoglycoside-resistant genes that encode various aminoglycoside-modifying enzymes (AME). Therefore, this study was conducted to compare the distribution of high-level aminoglycoside-resistant (HLAR) and multidrug-resistant (MDR) Enterococcus faecalis (E. faecalis) bacteria isolated from bulk tank milk in four dairy companies in Korea. Moreover, it analyzed the characteristics of their antimicrobial resistance genes and virulence factors. Among the 301 E. faecalis bacteria studied, 185 (61.5%) showed HLAR with no significant differences among the dairy companies. Furthermore, 129 (69.7%) of the 185 HLAR E. faecalis showed MDR without significant differences among companies. In contrast, HLAR E. faecalis from companies A, B, and C were significantly higher in resistance to the four classes than those in company D, which had the highest MDR ability against the three antimicrobial classes (p < 0.05). In addition, in the distribution of AME genes, 72 (38.9%) and 36 (19.5%) of the isolates carried both aac(6')Ie-aph(2″)-la and ant(6)-Ia genes, and the ant (6)-Ia gene alone, respectively, with significant differences among the companies (p < 0.05). In the distribution of virulence genes, the ace (99.5%), efa A (98.9%), and cad 1 (98.4%) genes were significantly prevalent (p < 0.05). Thus, our results support that an advanced management program by companies is required to minimize the dissemination of antimicrobial resistance and virulence factors.

High-Level Aminoglycoside Resistance in Human Clinical Klebsiella pneumoniae Complex Isolates and Characteristics of armA-Carrying IncHI5 Plasmids.

Aminoglycosides are important options for treating life-threatening infections. However, high levels of aminoglycoside resistance (HLAR) among Klebsiella pneumoniae isolates have been observed to be increasing frequently. In this study, a total of 292 isolates of the K. pneumoniae complex from a teaching hospital in China were analyzed. Among these isolates, the percentage of HLAR strains was 13.7% (40/292), and 15 aminoglycoside resistance genes were identified among the HLAR strains, with rmtB being the most dominant resistance gene (70%, 28/40). We also described an armA-carrying Klebsiella variicola strain KP2757 that exhibited a high-level resistance to all aminoglycosides tested. Whole-genome sequencing of KP2757 demonstrated that the strain contained one chromosome and three plasmids, with all the aminoglycoside resistance genes (including two copies of armA and six AME genes) being located on a conjugative plasmid, p2757-346, belonging to type IncHI5. Comparative genomic analysis of eight IncHI5 plasmids showed that six of them carried two copies of the intact armA gene in the complete or truncated Tn1548 transposon. To the best of our knowledge, for the first time, we observed that two copies of armA together with six AME genes coexisted on the same plasmid in a strain of K. variicola with HLAR. Comparative genomic analysis of eight armA-carrying IncHI5 plasmids isolated from humans and sediment was performed, suggesting the potential for dissemination of these plasmids among bacteria from different sources. These results demonstrated the necessity of monitoring the prevalence of IncHI5 plasmids to restrict their worldwide dissemination.

How do I manage a patient with enterococcal bacteraemia?

BACKGROUND: Enterococcal bacteraemia (EB) is common, particularly in the nosocomial setting, and its management poses a challenge for clinicians and microbiologists. OBJECTIVES: The aim was to summarize the more relevant features of EB and to provide a practical state-of-the-art on the topics that more directly affect its management. SOURCES: Pubmed articles from inception to 31 May 2020. CONTENT: The following topics are covered: epidemiological, clinical and microbiological characteristics and factors associated with prognosis of EB; diagnosis and work-up, including the use of echocardiography to rule out endocarditis; antibiotic management with special focus on antimicrobial resistance and complicated EB; and the role of infectious disease consultation and the use of bundles in EB. In addition, three clinical vignettes are presented to illustrate the practical application of the guidance provided, and major gaps in the current evidence supporting EB management are discussed. IMPLICATIONS: EB is associated with large burdens of morbidity and mortality, particularly among fragile and immunosuppressed patients presenting complicated bacteraemia due to multidrug-resistant enterococci. Most cases of EB are caused by Enterococcus faecalis, followed by E. faecium. EB often presents as polymicrobial bacteraemia. Rapidly identifying patients at risk of EB is crucial for timely application of diagnostic techniques and empiric therapy. Early alert systems and rapid diagnostic techniques, such as matrix-assisted desorption ionization-time of flight mass spectrometry, especially if used together with infectious disease consultation within bundles, appear to improve management and prognosis of EB. Echocardiography is also key in the work-up of EB and should probably be more extensively used, although its exact indications in EB are still debated. Multidisciplinary approaches are warranted due to the complexity and severity of EB.

Prevalence of High-Level Aminoglycoside Resistance and Genes Encoding Aminoglycoside-Modifying Enzymes in Enterococcus faecalis and Enterococcus faecium Isolated in a University Hospital in Tokyo.

High-level aminoglycoside resistance (HLAR) limits treatment options for invasive enterococcal infections. We examined the prevalence of HLAR, carriage of genes encoding aminoglycoside-modifying enzymes, and production of ß-lactamase using the disk diffusion method, polymerase chain reaction, and a nitrocefin-based test, respectively, in Enterococcus faecalis and Enterococcus faecium isolated from patients at a university hospital in Tokyo in 2010. Of the 100 E. faecalis isolates analyzed, 30 isolates had high-level resistance (HLR) to gentamicin, and 22 isolates had HLR to streptomycin. Of the 40 E. faecium isolates analyzed, 9 isolates had HLR to gentamicin, and 9 isolates had HLR to streptomycin. Of the 39 gentamicin-HLR enterococcal isolates, 24 isolates were non-HLR to streptomycin. All 39 isolates with HLR to gentamicin as well as 19 of 101 without HLR carried aac(6')-Ie-aph(2'')-Ia. Carriage of ant(6')-Ia was confirmed in 25 of 31 streptomycin-HLR isolates. Production of ß-lactamase was documented in none of the E. faecalis and E. faecium isolates. Whole-genome sequencing analysis revealed that all but one E. faecalis isolate that carried aac(6')-Ie-aph(2'')-Ia and ant(6')-Ia belonged to sequence type (ST) 4 (n = 8), ST16 (n = 4), or ST179 (n = 9). Nevertheless, most of the pairs of isolates had > 10 single-nucleotide polymorphisms even among the isolates of the same ST.

Assessment of Tedizolid In Vitro Activity and Resistance Mechanisms against a Collection of Enterococcus spp. Causing Invasive Infections, Including Isolates Requiring an Optimized Dosing Strategy for Daptomycin from U.S. and European Medical Centers, 2016 to 2018.

High-level aminoglycoside resistance was noted in 30.0% of Enterococcus faecalis and 25.2% of Enterococcus faecium isolates. Only 3.3% and 2.1% of E. faecalis isolates had elevated daptomycin MIC (≥2 mg/liter) and vancomycin resistance, respectively. In contrast, 37.4% to 40.3% of E. faecium isolates exhibited these phenotypes. Tedizolid inhibited 98.9% to 100.0% of enterococci causing serious invasive infections, including resistant subsets. Oxazolidinone resistance was mainly driven by G2576T; however, optrA and poxtA genes were also detected, including poxtA in the United States and Turkey.

Detection of antibiotic resistance profiles and aminoglycoside-modifying enzyme (AME) genes in high-level aminoglycoside-resistant (HLAR) enterococci isolated from raw milk and traditional cheeses in Turkey.

The aim of this study was isolation and identification of the high-level aminoglycoside-resistant (HLAR) enterococci in raw milk and dairy products and to analyze their antibiotic resistance and the presence of aminoglycoside-modifying enzyme (AME) genes. A total of 59 HLAR enterococci were isolated from raw milk and traditional cheese samples. Thirty-nine of the 59 HLAR enterococci were isolated on streptomycin-containing agar medium, while the other 20 HLAR strains were isolated on gentamicin containing agar medium. The 59 HLAR enterococci were identified as 26 E. faecalis (44.07%), 18 E. faecium (30.51%), 13 E. durans (22.03%), and two E. gallinarum (3.39%) by species-specific PCR. Disk diffusion tests showed that teicoplanin were the most effective antibiotics used in this study, while 89.83% of isolates were found to be resistant to tetracycline. High rates of multiple antibiotic resistance were detected in HLAR isolates. Minimum inhibitory concentration (MIC) values of HLAR enterococci against streptomycin and gentamicin were found in the range of 64 to > 4096 µg/mL. Forty-seven (79.66%) of the 59 HLAR enterococci were found to be both high-level streptomycin-resistant (HLSR) and high-level gentamicin-resistant (HLGR) by MIC tests. However, no correlation was found between the results of the disk diffusion and MIC tests for gentamicin and streptomycin in some HLAR strains. The aph(3')-IIIa (94.92%) was found to be most prevalent AME gene followed by ant(4')-Ia (45.76%), ant(6')-Ia (20.34%) and aph(2'')-Ic (10.17%). None of the isolates contained the aac(6')-Ie-aph(2'')-Ia, aph(2'')-Ib or aph(2'')-Id genes. None of the AME-encoding genes were identified in E. durans RG20.1, E. faecalis RG22.4, or RG26.1. In conclusion, HLAR enterococci strains isolated in this study may act as reservoirs in the dissemination of antibiotic resistance genes.

High-level aminoglycoside resistance and distribution of aminoglycoside resistance genes among Enterococcus spp. clinical isolates in Tehran, Iran.

OBJECTIVES: Enterococci have gained attention during the past decade as important nosocomial pathogens. Their increasing prevalence has been paralleled by the occurrence of multidrug-resistant and high-level aminoglycoside-resistant strains. This study isolated Enterococcus spp. from hospital samples and determined their antibiotic resistance profile, focusing on aminoglycosides, and associated resistance mechanisms. METHODS: A total of 195 enterococci from hospital samples in Tehran were studied. Isolates were identified by biochemical reactions. Antimicrobial resistance was determined by disk diffusion. The vancomycin MIC for vancomycin-resistant isolates was determined by agar dilution. Detection of aminoglycoside resistance genes and intI1 and intI2 gene was performed by PCR. RESULTS: The majority of isolates were Enterococcus faecalis (65.1%), followed by Enterococcus faecium (31.8%), Enterococcus gallinarum (2.6%) and Enterococcus solitarius (0.5%). According to antibiogram results, 42.1% of isolates were high-level gentamicin-resistant (HLGR) and 40.5% were high-level streptomycin-resistant (HLSR). There was a high prevalence of aac(6')-Ie-aph(2")-Ia (96.3%) among HLGR isolates. ant(6)-Ia and aadA were identified in 93.7% and 64.6% of HLSR isolates, respectively. aph(2'')-Ic was detected in 7 isolates (3.6%) and aph(2'')-Ib in only 4 isolates (2.1%); no isolates harboured aph(2'')-Id, intI1 or intI2. CONCLUSION: Multidrug resistance was higher among HLGR and HLSR isolates compared with non-HLGR and non-HLSR isolates, which may result in limited treatment options. More than 50% of isolates were susceptible to aminoglycosides, thus correct identification in clinical laboratories and administration of these antibiotics can result in decreased used of antibiotics such as vancomycin and linezolid and help to reduce the emergence of resistance to these drugs.

Study of vancomycin and high-level aminoglycoside-resistant Enterococcus species and evaluation of a rapid spot test for enterococci from Central Referral Hospital, Sikkim, India.

BACKGROUND: Enterococcus is an important pathogen, and with its emergence of resistance to multiple antimicrobials, the management of infection is becoming increasingly difficult. AIM: The aim of the study is to determine the prevalence, antibiotic resistance, and risk factors associated with enterococcal infection or colonization. MATERIALS AND METHODS: In this prospective study, samples from inpatients were screened for resistant enterococci. Antibiotic susceptibility testing was performed using the disc diffusion method and minimum inhibitory concentration by the agar dilution method. A modification of a test tube method of sodium chloride-esculin hydrolysis to a spot test was evaluated for its rapidity and reliability in the presumptive diagnosis of enterococci. STATISTICAL ANALYSIS USED: Fisher's exact test was used for continuous (Student's t-test) and categorical variables. Multivariate analysis was performed with logistic regression using IBM SPSS 20.0 software (Armonk, NY, USA). RESULTS: Enterococcus species were isolated from 182 samples: Enterococcus faecalis (68.7%), Enterococcus faecium (20.9%), Enterococcus gallinarum (6%), and Enterococcus durans (4.4%). Maximum resistance was to ciprofloxacin (59.3%) and least to linezolid (0.5%). The isolation rate of vancomycin-resistant enterococci (VRE) was 13.7%; 30.2% and 20.9% were of high-level gentamicin and streptomycin, respectively. All 182 Enterococcus species gave positive results within 30-60 min by the rapid spot test. CONCLUSIONS: Overall, high-level aminoglycoside resistance (HLAR) was observed more than glycopeptide resistance. Surveillance strategies need to be upgraded and implemented in order to prevent the emergence and further spread of not only VRE but also HLAR enterococci in the hospital. The spot test gave reliable and rapid results in presumptive identification of enterococci.

High Prevalence of the aac(6')-Ie-aph(2'')-Ia Gene in Hospital Isolates of Enterococcus faecalis Co-Resistant to Gentamicin and Penicillin.

Objectives: This study aimed to characterize the molecular mechanism of resistance to gentamicin among penicillin-resistant, ampicillin-susceptible Enterococcus faecalis (PRASEF) isolates by investigating the presence of the aac(6')-Ie-aph(2'')-Ia gene. The co-resistance to antimicrobials of other classes was also evaluated. Results: Among the 151 isolates evaluated, 70 were PRASEF and 81 were penicillin-susceptible and ampicillin-susceptible E. faecalis (PSASEF). No ß-lactamase producing isolate was detected. Eighty-three (55.0%) and 35 (23.2%) out of the 151 E. faecalis isolates showed high-level gentamicin resistance (HLGR) and high-level streptomycin resistance (HLSR) phenotypes. However, a significantly higher rate of PRASEF (88.6%) showed HLGR phenotype in comparison with PSASEF (23.5%) (p < 0.01). Conversely, a significantly lower rate of PRASEF (14.3%) showing HLSR was observed in comparison with PSASEF (30.9%) (p = 0.02). The prevalence of isolates displaying multidrug resistance (MDR) phenotype was significantly higher (p < 0.01) in the group of PRASEF (81.4%) than in PSASEF (18.6%). The majority of PSASEF (61.9%) and PRASEF (90.3%) isolates showing HLGR phenotype was harboring the aac(6')-Ie-aph(2'')-Ia gene, which encodes a bifunctional enzyme that inactivates all aminoglycosides except streptomycin. Conclusion: The aac(6')-Ie-aph(2'')-Ia gene was prevalent among the Brazilian PRASEF isolates that usually exhibit co-resistance to gentamicin and to multiple other drugs.

High level aminoglycoside resistant enterococci in hospital-acquired urinary tract infections in Mansoura, Egypt.

INTRODUCTION: Resistance to different antimicrobial agents is increasing in enterococci and effective treatment represents a major health concern. The aim of this study was to determine the antimicrobial resistance patterns and the frequency of high level aminoglycoside resistance (HLAR) among enterococci. METHODS: A total of 80 enterococcal isolates, (73 Enterococcus faecalis, 7 Enterococcus faecium) were collected from patients with hospital acquired urinary tract infections (UTI) at Mansoura University hospitals in Egypt. Antimicrobial susceptibility testing was performed via the disc diffusion method. PCR was used for identification of species and detection of aminoglycoside-modifying enzymes genes (AME). RESULTS: All enterococcal isolates were sensitive to vancomycin and linezolid. Fifty-three isolates exhibited HLAR. Our results show that HLAR was mediated by the presence of multiple AMEs genes. The aac(6')-Ie-aph(2')-Ia gene was associated with aph(3')-IIIa and ant(6)-Ia gene in 69% of HLAR isolates. CONCLUSION: This study showed that enterococci isolated from hospital acquired UTI were resistant to multiple antibiotics. Furthermore, the frequency of high level gentamicin resistance (HLGR) was higher than high level of streptomycin resistance (HLSR). The most common AME genes were aph(3')-IIIa and ant(6)-Ia followed by aac(6')-Ie-aph(2')-Ia.

Linezolid and Vancomycin Resistant Enterococci: A Therapeutic Problem.

INTRODUCTION: Enterococci are recognized as opportunistic pathogens, as well as commensals in both humans and animals. They are an important cause of nosocomial infections, difficult to treat, as the organism have intrinsic and acquired resistance to many antibiotics. AIM: To isolate and identify clinically relevant Enterococcus up to species level from all the clinical samples processed in the microbiology laboratory and also to study their resistance pattern. MATERIALS AND METHODS: A prospective study was carried out for a period of one year from May 2014 to April 2015 at the Department of Microbiology, Subharti Medical College, Meerut, Uttar Pradesh, India. A total of 200 isolates of Enterococcus species from 15342 clinical samples obtained from IPD/OPD patients irrespective of age, having suspicion of bacterial infection were processed in the microbiology laboratory. Identification was done with standard biochemical methods. Antibiotic susceptibility testing was done on Muller Hilton agar plate by Kirby Bauer disc diffusion method. High-Level Gentamycin Resistance (HLGR) and High-Level Streptomycin Resistance (HLSR) were further confirmed by Agar dilution method and Broth microdilution method. Vancomycin and linezolid resistance was further confirmed by Agar dilution method and MIC was calculated by using VITEK 2, Biomerieux. All methodology was followed as per Clinical and Laboratory Standards Institute (CLSI) M100-S 24, 2014 guidelines. RESULTS: E. faecalis (n=169, 84.5%) was the predominant species isolated, followed by E. faecium (n=27, 13.5%) and E. casseliflavus (n=4, 2%). A total of 25 (12.5%) isolates were HLGR, 13 (6.5%) isolates were HLSR and 62 (31%) isolates were HLGR+HLSR. Vancomycin resistance was found in 14 (7%) isolates of which 11 (78.5%) were Van A and 3 (21.4%) were Van B, detected phenotypically as per relative MIC of vancomycin and teicoplanin. Linezolid resistance was seen in 4 (2%) of isolates which were vancomycin as well as high-level gentamycin and high-level streptomycin resistant. CONCLUSION: Isolation of Enterococcus species resistant to most of the higher antibiotics like vancomycin and linezolid, with high prevalence of High-Level Aminoglycoside Resistance (HLAR), from hospitalized patients is a major concern as such isolates have limited or no therapeutic option.

Conjugal transfer of aac(6')Ie-aph(2″)Ia gene from native species and mechanism of regulation and cross resistance in Enterococcus faecalis MCC3063 by real time-PCR.

High level aminoglycoside resistance (HLAR) in the lactic acid bacteria (LAB) derived from food animals is detrimental. The aim of this study was to investigate the localization and conjugal transfer of aminoglycoside resistance genes, aac(6')Ie-aph(2″)Ia and aph(3')IIIa in different Enterococcus species. The cross resistance patterns in Enterococcus faecalis MCC3063 to clinically important aminoglycosides by real time PCR were also studied. Southern hybridization experiments revealed the presence of aac(6')Ie-aph(2″)Ia and aph(3')IIIa genes conferring HLAR in high molecular weight plasmids except in Lactobacillus plantarum. The plasmid encoded bifunctional aac(6')Ie-aph(2″)Ia gene was transferable from Enterococcus avium (n = 2), E. cecorum (n = 1), E. faecalis (n = 1) and Pediococcus lolii (n = 1) species into the recipient strain; E. faecalis JH2-2 by filter mating experiments thus indicating the possible risks of gene transfer into pathogenic strains. Molecular analysis of cross resistance patterns in native isolate of E. faecalis MCC3063 carrying aac(6')Ie-aph(2″)Ia and aph(3')IIIa gene was displayed by quantification of the mRNA levels in this study. For this, the culture was induced with increasing concentrations of gentamicin, kanamycin and streptomycin (2048, 4096, 8192, 16384 µg/mL) individually. The increasing concentrations of gentamicin and kanamycin induced the expression of the aac(6')Ie-aph(2″)Ia and aph(3')IIIa resistance genes, respectively. Interestingly, it was observed that induction with streptomycin triggered a significant fold increase in the expression of the aph(3')IIIa gene which otherwise was not known to modify the aminoglycoside. This is noteworthy as streptomycin was found to confer cross resistance to structurally unrelated kanamycin. Also, expression of the aph(3')IIIa gene when induced with streptomycin, revealed that bacteria harbouring this gene will be able to overcome streptomycin bactericidal action at specific concentrations. HLAR in E. faecalis MCC3063 may be due to the combined expression of both the aac(6')Ie-aph(2″)Ia and aph(3')IIIa genes which could be therapeutically challenging. A combined expression of both the genes in E. faecalis MCC3063 may yield HLAR which could be therapeutically challenging. The study highlights the significant alterations in the mRNA expression levels of aac(6')Ie-aph(2″)Ia and aph(3')IIIa in resistant pathogens, upon exposure to clinically vital aminoglycosides.

Antimicrobial resistance in Enterococcus strains isolated from healthy domestic dogs.

Enterococci are opportunistic bacteria that cause severe infections in animals and humans, capable to acquire, express, and transfer antimicrobial resistance. Susceptibility to 21 antimicrobial agents was tested by the disk diffusion method in 222 Enterococcus spp. strains isolated from the fecal samples of 287 healthy domestic dogs. Vancomycin and ampicillin minimum inhibitory concentrations (MICs) and high-level aminoglycoside resistance (HLAR) tests were also performed. Isolates showed resistance mainly to streptomycin (88.7%), neomycin (80.6%), and tetracycline (69.4%). Forty-two (18.9%) isolates showed an HLAR to streptomycin and 15 (6.7%) to gentamicin. Vancomycin and ampicillin MIC values showed 1 and 18 resistant strains, respectively. One hundred and thirty-six (61.2%) strains were classified as multidrug resistant and six (2.7%) strains as possibly extensively drug-resistant bacteria. Enterococcus faecium and Enterococcus faecalis were the most prevalent antimicrobial resistant species. Companion animals, which often live in close contact with their owners and share the same environment, represent a serious source of enterococci resistant to several antibiotics; for this reason, they may be a hazard for public health by providing a conduit for the entrance of resistance genes into the community.

The prevalence of aminoglycoside-modifying enzyme and virulence genes among enterococci with high-level aminoglycoside resistance in Inner Mongolia, China

ABSTRACT This study highlights the prevalence of aminoglycoside-modifying enzyme genes and virulence determinants among clinical enterococci with high-level aminoglycoside resistance in Inner Mongolia, China. Screening for high-level aminoglycoside resistance against 117 enterococcal clinical isolates was performed using the agar-screening method. Out of the 117 enterococcal isolates, 46 were selected for further detection and determination of the distribution of aminoglycoside-modifying enzyme-encoding genes and virulence determinants using polymerase chain reaction -based methods. Enterococcus faecium and Enterococcus faecalis were identified as the species of greatest clinical importance. The aac(6')-Ie-aph(2")-Ia and ant(6')-Ia genes were found to be the most common aminoglycoside-modifying enzyme genes among high-level gentamicin resistance and high-level streptomycin resistance isolates, respectively. Moreover, gelE was the most common virulence gene among high-level aminoglycoside resistance isolates. Compared to Enterococcus faecium, Enterococcus faecalis harbored multiple virulence determinants. The results further indicated no correlation between aminoglycoside-modifying enzyme gene profiles and the distribution of virulence genes among the enterococcal isolates with high-level gentamicin resistance or high-level streptomycin resistance evaluated in our study.

The prevalence of aminoglycoside-modifying enzyme and virulence genes among enterococci with high-level aminoglycoside resistance in Inner Mongolia, China.

This study highlights the prevalence of aminoglycoside-modifying enzyme genes and virulence determinants among clinical enterococci with high-level aminoglycoside resistance in Inner Mongolia, China. Screening for high-level aminoglycoside resistance against 117 enterococcal clinical isolates was performed using the agar-screening method. Out of the 117 enterococcal isolates, 46 were selected for further detection and determination of the distribution of aminoglycoside-modifying enzyme-encoding genes and virulence determinants using polymerase chain reaction -based methods. Enterococcus faecium and Enterococcus faecalis were identified as the species of greatest clinical importance. The aac(6')-Ie-aph(2″)-Ia and ant(6')-Ia genes were found to be the most common aminoglycoside-modifying enzyme genes among high-level gentamicin resistance and high-level streptomycin resistance isolates, respectively. Moreover, gelE was the most common virulence gene among high-level aminoglycoside resistance isolates. Compared to Enterococcus faecium, Enterococcus faecalis harbored multiple virulence determinants. The results further indicated no correlation between aminoglycoside-modifying enzyme gene profiles and the distribution of virulence genes among the enterococcal isolates with high-level gentamicin resistance or high-level streptomycin resistance evaluated in our study.

High-level aminoglycoside resistance in Enterococcus faecalis and Enterococcus faecium causing invasive infection: Twelve-year surveillance in the Minami Ibaraki Area.

We examined prevalence of high-level aminoglycoside resistance (HLAR) in Enterococcus faecalis and Enterococcus faecium causing invasive infection in the Minami Ibaraki Area. Ten strains of both species each, recovered from the blood or the cerebrospinal fluid between 2003 and 2014, were randomly selected every year. High-level resistance to gentamicin (HLR-GM) and streptomycin (HLR-SM) was detected in 34% (41 of 120 strains) and 18% (21) of E. faecalis and 9% (11) and 39% (48) of E. faecium, respectively. In comparisons of the proportions among three four-year periods, HLR-SM among E. faecium was significantly lower in the 2011-2014 period. All strains with HLR-GM were positive for the aac(6')-Ie-aph(2″)-Ia gene. The ant(6')-Ia gene was detected in all with HLR-SM except for one E. faecalis strain. The present study showed that prevalence of HLR-GM among E. faecalis and E. faecium causing invasive infection in this area was nearly equivalent to that described in previous studies in Japan and that proportions of strains with HLAR did not vary during the study period except for that of HLR-SM among E. faecium.

Clinico-Microbiological Investigation of Catheter Associated Urinary Tract Infection by Enterococcus faecalis: vanA Genotype.

Prolonged hospitalization and exposure to third generation cephalosporins are reported to facilitate the acquisition and colonization of Vancomycin Resistant Enterococci (VRE). Though VRE is not uncommon in India, urinary tract infection with a vanA genotype is a cause of serious concern as VRE co-exhibit resistance to aminoglycosides. In India, majority of the VRE isolates recovered from hospitalized patients include Enterococcus faecium. We report a case of catheter associated urinary tract infection by an endogenous, multidrug resistant E. faecalis of vanA genotype following prolonged hospitalization, ICU stay, catheterisation and exposure to 3G cephalosporin and metronidazole. The patient responded to linezolid therapy.

Emergence of multidrug resistant enterococci at a tertiary care centre.

BACKGROUND: Enterococci have assumed great clinical importance because of their increasing resistance to various antimicrobial agents. Thus, knowledge about the antibiogram of these multidrug resistant isolates is of utmost importance in formulating an effective antibiotic policy to treat these infections and reducing the morbidity and mortality. Aim of this study was to assess the antimicrobial resistance pattern of enterococci and determine the prevalence of multidrug resistance among them. METHODS: This cross sectional study was carried out from August 2011 to February 2014, in which 200 non-repetitive clinical isolates of enterococci were included. Antimicrobial susceptibility testing was done by disc diffusion method. Minimum inhibitory concentration (MIC) of gentamicin, streptomycin, vancomycin, teicoplanin and linezolid was determined by E-test method. RESULTS: The prevalence of multidrug resistance among enterococcal isolates was found to be 63%. Varying levels of resistance was seen to various antibiotics. Most of the isolates were resistant to penicillin (95%), ampicillin (95%) and cotrimoxazole (90%). High level aminoglycoside resistance (HLAR) and glycopeptide resistance was seen in 39% and 14% isolates respectively. Only 4 isolates (2%) were found to be resistant to linezolid. CONCLUSION: The prevalence of multidrug resistance among enterococci was found to be 63%, the resistance being more common in Enterococcus faecium as compared to Enterococcus faecalis. The study highlights the emergence and increased prevalence of multidrug resistant enterococci which pose a serious therapeutic challenge.