Drug resistance of 100 clinical strains of Enterococcus spp

The aim of this study was to evaluate the drug susceptibility of 100 Enterococcus spp. strains isolated from patients hospitalized in State Clinical Hospital No 1 in Warsaw. All strains were identified (API 20 STREP) and their susceptibility to antibiotics was tested (ATB STREP) in automatic ATB system. Additionally, PYRase activity, beta-lactamase production (in nitrocefin test), MICs for vancomycin and teicoplanin (E test), HLAR--high level aminoglycoside resistance and susceptibility to vancomycin, teicoplanin, piperacillin and piperacillin/tazobactam (disc diffusion method) were determined. E. faecalis ATCC 29212 was used as the control strain. Fifty E. faecalis, 45 E. faecium, 2 E. casseliflavus, 2 E. durans and 1 E. avium strain were cultured. All strains were PYRase-positive and beta-lactamase-negative. Ten isolates demonstrated intermediate susceptibility to vancomycin (6--E. faecalis and 4--E. faecium). One E. faecalis strain was intermediately susceptible to both glycopeptides. One E. casseliflavus strain showed low-level resistance to vancomycin, but this strain was susceptible to teicoplanin--phenotype Van C. HLAR strains were found among 31 E. faecalis and 40 E. faecium strains. 48 E. faecalis strains were susceptible to piperacillin and 49 to piperacillin/tazobactam. Whereas, 41 E. faecium were resistant to both these drugs. Thirty six per cent of isolates were resistant to penicillin and ampicillin, 73% to erythromycin, 87% to tetracycline, 89% to lincomycin and 56% to nitrofurantoin. Some discrepancies were noticed between the results of different methods applied for susceptibility testing--ATB system, E test and disc diffusion. These discrepancies concerned HLAR detection and susceptibility to glycopeptides determination. The best methods were: disc-diffusion for HLAR detection and E test for determination of resistance to vancomycin and teicoplanin. Increasing resistance to antimicrobial agents is observed in clinical Enterococcus spp. isolates cultured in our laboratory, especially in E. faecium strains. It is necessary to control the dissemination of multiresistant Enterococcus spp. strains in hospital wards.     

Antimicrobial resistance patterns among different Escherichia coli isolates in the Kingdom of Saudi Arabia

Antimicrobial resistance patterns among different Escherichia coli isolates in the Kingdom of Saudi Arabia. This study aimed to investigate the patterns of antimicrobial resistance in E. coli isolated from different samples, and to identify potential pathogenic isolates in Riyadh, Kingdom of Saudi Arabia (KSA). In total, 51 bacterial isolates were recovered from 113 samples of human urine, food (raw meat, raw chicken, raw egg surface, and fresh vegetables), water, and air. Twenty-four E. coli isolates were tested for susceptibility to 26 antibiotics. The air sample isolates were most resistant to amoxicillin, ampicillin, amoxicillin/clavulanic acid, amoxicillin/sulbactam, piperacillin/tazobactam, cefalotin, cefuroxime, cefoxitin, cefixime, nitrofurantoin, and trimethoprim/sulfamethoxazol. The isolates from vegetable samples were resistant to amoxicillin, ampicillin, amoxicillin/clavulanic acid, amoxicillin/sulbactam, cefalotin, cefuroxime, cefoxitin, and cefixime. By contrast, the isolates from the water samples were resistant only to amoxicillin and ampicillin. The isolates from the human urine samples were most frequently resistant to norfloxacin (80%) followed by amoxicillin and ampicillin (70%), trimethoprim/sulfamethoxazole (55%), ciprofloxacin and ofloxacin (50%), cefalotin (30%), cefuroxime, cefixime and cefotaxime (25%), ceftazidime, ceftriaxone, cefepime and aztreonam (20%), amoxicillin/clavulanic acid, piperacillin/tazobactam and gentamicin (10%), and amoxicillin/sulbactam and cefoxitin (5%). Almost all (23/25, 95.8%) (n = 23) of the isolates were multi-drug resistant (MDR) (i.e., resistant to 3 or more classes of antibiotics), and 16.7% (n = 4) of those were positive for extended spectrum β-lactamase (ESBL). Of the 4 ESBL-producers, 3 were positive for blaCTX_-M-15 and blaCTX_-M1_group, 2 were positive for blaCMY_-2, and 1 each was positive for blaCTX_{-M-2 group,} blaSHV, and blaOXA_-47. The quinolone resistance gene qnrS was detected in 25% (n = 6) of the E. coli strains isolated from urine (N = 5) and air (N = 1) samples. The considerable number of antimicrobial resistance genes detected among E. coli isolates tested here is alarming and should raise public health concern.     

Infectivity and resistance to antibiotics of bacterial strains isolated from patients hospitalised in intensive care units

The aim of this study was to evaluate the frequency of isolation and antimicrobial resistance testing of bacterial strains isolated from clinical specimens from patients hospitalized in three Intensive Care Units in Wroc?aw. The susceptibility of bacteria (107 strains) to selected antibiotics was determined. The results clearly show that non-fermentative rods were identified as the main agents causing pneumonia (58% of isolates). The second commonest pathogens were Gram-positive cocci (29%). The P. aeruginosa and E. cloacae strains were resistant to ampicillin, amoxicillin/clavulanate, cefuroxime and cefotaxime. All isolates of A. baumanii were susceptible only to imipenem. The rods of K. pneumoniae and E. coli were resistant to ampicillin, about 55% strains of both bacteria were sensitive to other antibiotics, except piperacillin/tazobactam, imipenem and ciprofloxacin. About 90% of methicillin resistant S. epidermidis strains were resistant to all antibiotics, except vancomycin (100% isolates were sensitive). ESBL were detected among E. cloace, K. pneumoniae and E. coli. We found P. aeruginosa rods producing MBL.     

Antimicrobial susceptibility of microorganisms isolated from urine of pediatric ward patients

The study was an analysis of the frequency of urine bacterial isolation in hospitalized children as well as an evaluation of their susceptibility to antibiotics used in urinary tract infections (UTI). The analysis focused on microbiological urine tests carried out between January 2006 and December 2008. Altogether, 311 strains were obtained, of which E. coli (50.8%) and E. faecalis (13.5%) were the most frequently isolates. The highest percentage of Enterobacteriaceae were sensitive to ceftazidime (92%); to a lesser degree to amoxicillin/clavulanic acid (85%), to trimethoprim/sulfamethoxazole (84%), to nitrofurantoin (82%), to cefuroxime (81%), to cefalotin (66%) whereas only 24% were sensitive to ampicillin. ESBLs were produced by 8% of all Enterobacteriaceae strains. P. aeruginosa strains were totally sensitive to ceftazidime; over 90% - to piperacillin and aminoglycosides, and 77% to carbenicillin. Staphylococci manifested 100% sensitivity to nitrofurantoin. Only 20% of S. aureus were sensitive to trimethoprim/sulfamethoxazole and to trimethoprim; in the case of S. epidermidis: 83% and 67% respectively. No resistant strains were found among S. agalactiae and E. faecalis. E. faecium strains, in turn, were resistant to ampicillin and often to nitrofurantoin (64%), to vancomycin (VanB; 45%) and to high aminoglycoside concentrations (HLAR; 45%).     

Surveillance and trends of antimicrobial resistance among clinical isolates of anaerobes in Kuwait hospitals from 2002 to 2007

The susceptibility trends for all anaerobes processed by the Anaerobe Reference Laboratory against various antibiotics were determined by using data for 2557 isolates referred by all government hospitals in Kuwait from 2002 to 2007. MIC were determined for the following anti-anaerobic antibiotics: amoxicillin–clavulanic acid, clindamycin, imipenem, meropenem, metronidazole, penicillin, piperacillin, piperacillin–tazobactam and vancomycin (for Gram-positive anaerobes only), using E-test method. The commonest isolates were Bacteroides fragilis (36.8%), followed by Peptostreptococcus spp. (21.9%), Bacteroides ovatus (15.5%) and Prevotella bivia (12.1%). In addition, Prevotella oralis and other Bacteroides spp. represented 8.5% and 8.1% of total number of isolates, respectively. Resistance rate varied among the antimicrobial agents and the species tested. The β-lactams, with the exception of penicillin, were the most active drugs. Piperacillin–tazobactam was the only antimicrobial agent to which all the isolates were uniformly susceptible. Imipenem and metronidazole were highly active with resistance rate of only <5% recorded against most isolates. However, 42.8, 55.8 and 9.3% of Clostridium difficile isolates were resistant to imipenem, clindamycin and meropenem, respectively. It is noteworthy that from 2002 to 2007, there was a gradual increase in resistance rates to clindamycin, amoxicillin–clavulanic acid and piperacillin among B. fragilis. Periodic surveillance of antibiotic resistance among the anaerobic bacteria is recommended as a guide to empiric antibiotic use and formulation of guideline for appropriate choice of antimicrobial therapy in anaerobic infections.     

Increased frequency of integrons and β-lactamase-coding genes among extraintestinal Escherichia coli isolated with a 7-year interval

We analyzed the level of antimicrobial resistance, and the presence of integrons and β-lactamase-coding genes in 69 clinically relevant Escherichia coli strains originating from extraintestinal infections isolated in 1999–2001 and 2008–2010. Comparison of the two groups showed significant differences in drug resistance frequency, and the presence of integron and β-lactamase-coding genes. The frequency of resistance to all antimicrobials beside imipenem, streptomycin, piperacillin/tazobactam, and sulfamethoxazole increased significantly, especially towards aminoglycosides, β-lactams and fluoroquinolones. Similarly, we noticed an increase in the number of strains with integrons from 31.6 to 80.7 %. The presence of integrase genes was associated with elevated frequency of resistance to each antimicrobial tested besides imipenem, piperacillin/tazobactam and ceftazidime. The presence of integrons was also associated with multidrug resistance phenotype. The genetic content of integrons comprised genes determining resistance toward aminoglycosides, sulfonamides and trimethoprim. Moreover, we noticed a significant increase in the frequency of bla _CTX-M β-lactamases, with appearance of bla _CTX-M-15 variant and newer plasmid-encoded β-lactamases like CMY-15 and DHA. The emergence of strains resistant to several classes of antimicrobials and carrying integrons, ESBL and AmpC β-lactamase-coding genes may predict the spread of isolates with limited treatment options.     

Profile and multidrug resistance determinants of Chryseobacterium indologenes from seawater and marine fauna

The aim of this study was to investigate the prevalence and genetic basis of multidrug resistance in Chryseobacterium indologenes from seawater and marine invertebrates used for human consumption, in Ka?tela Bay, Adriatic Sea, Croatia. Out of 16 samples of seawater, Mediterranean mussel (Mytilus galloprovincialis Lam.), Rayed Mediterranean limpets (Patella caerulea L.) and Purple sea urchins (Paracentrotus lividus Lam.) collected, 15 were positive for C. indologenes. In total, 41 isolates were randomly selected and tested for antibiotic susceptibility by disc-diffusion and broth microdilution methods. PCR was used to detect alleles encoding extended-spectrum (ESBLs) and metallo-β-lactamases (MBLs). The clonality of β-lactamase-producing strains was evaluated by random amplified polymorphic DNA (RAPD) analysis. All C. indologenes isolates showed multiple resistance to at least 9 out of 16 antibiotics tested. Lowest resistance rates were found for piperacillin (9.7 %) and ciprofloxacin (24.4 %), whereas only piperacillin/tazobactam and trimethoprim/sulfamethoxazole showed 100 % activity. More than half of isolates carried bla _IND-type gene, including 2 isolates carrying bla _IND-2 and 21 carrying bla _IND-7, that was identified as a major MBL genotype in isolates from Adriatic Sea. RAPD typing of IND-producing isolates revealed 6 major groups with no predominant clone in population. The presence of multidrug resistant and IND-producing C. indologenes in marine environment, including marine fauna, pose a risk for transmitting this opportunistic pathogen to humans through recreation or consummation of seafood. In addition, the antibiotic susceptibility test results have practical relevance for empirical treatment of C. indologenes infections.     

Molecular characterization of antibiotic resistance in clinical Salmonella typhi isolated in Ghana

Fifty-eight clinical Salmonella typhi strains isolated from patients suspected of suffering from typhoid fever were obtained at the Korle-Bu Teaching Hospital and the Noguchi Memorial Institute for Medical Research, both located in Ghana, Africa. Each isolate was examined for susceptibility to ampicillin, chloramphenicol, streptomycin, tetracycline, and trimethoprim/sulfamethoxazole by the disk diffusion assay. Five of the isolates were resistant to all five antibiotics while 10 isolates were resistant to ampicillin, chloramphenicol, and trimethoprim/sulfamethoxazole, which are considered 'first line' antibiotics in the treatment of typhoid fever. Thirty-four isolates were resistant to at least one of the antibiotics tested and 62% of these isolates possessed conjugable plasmids belonging to incompatibility group IncHI. Ninety percent of the conjugable plasmids conferred a multiple drug-resistant phenotype on the strains harboring them. Additionally, 14 strains contained plasmids that were transformable and six of them encoded multiple drug resistance. Our findings indicate that multiple drug resistance to the 'first line' antibiotics in S. typhi may be more prevalent in Africa than previously thought.     

Physiological characteristics ofClostridium bifermentans selectively isolated from California desert tortoise

A selective agar medium containing cycloserine (250 mg/L), sulfamethoxazole (76 mg/L), and trimethoprim (4 mg/L) was used for isolation ofClostridium bifermentans from the intestinal contents of California desert tortoise. Typical lecithinase positive colonies that appeared on the plates, were biochemically characterized with the API 20A System and a conventional procedure. The susceptibility of the isolates to 12 antimicrobial agents was determined by the broth microtitration technique using the ANA MIC System.C. bifermentans strains were shown to be highly susceptible to cefoxitin, cefotetan, ceftizoxime, cefotaxime, chloramphenicol, clindamycin, penicillin, metronidazole, piperacillin, ticarcillin, and mezlocillin. Less than 10% of the isolates exhibited resistance to tetracycline. All strains were found to be highly resistant to cycloserine, sulfamethoxazole, and trimethoprim. The GLC analysis of the culture supernatants for volatile fatty acids revealed the presence of formic, acetic, isobutyric, butyric, isovaleric, and isocaproic acids.     

Synergistic effect of polyethylenimine plus piperacillin/tazobactam on clinicalPseudomonas aeruginosa isolates resistant to piperacillin/tazobactam

We investigated thein vitro susceptibility of piperacillin/tazobactam in association with polyethylenimine (PEI), a synthetic polycation polymer, against 24 clinical isolates ofPsedomonas aeruginosa resistant to piperacillin/tazobactam. The strains were isolated from patients with respiratory tract infections. MIC of piperacillin/tazobactam was determined by agar dilution method in accordance with NCCLS methodology. An inoculum of 10⁴ CFU/spot with or without PEI (250 nM final concentration) was used. Killing curves were performed for 3 piperacillin/tazobactam resistant strains chosen for their different range of MIC values to piperacillin/tazobactam (128, 256 and 512 mg/L, respectively). Analyses were performed in duplicate using a concentration of antibiotic of 16 mg/L. The addition of PEI (250 nM) increased the susceptibility of piperacillin/tazobactam (from 8 to 32 folds) in all the strains tested. Moreover, in all the strains tested piperacillin/tazobactam in association with PEI showed a bactericidal activity before 6 h. Intrinsic resistance ofP. aeruginosa affects susceptibility to penicillins and is believed to entail broad spectrum impermeability. Different theories suggest that some degree of structure or organization exists in the periplasm ofP. aeruginosa and that this modulates both the level of intrinsic, impermeability-determined resistance expressed by a strain and the efficiency with which the β-lactamase can contact and destroy the incoming β-lactamase molecules. The explanation of this behaviour could be that PEI determines an increased permeability of the outer membrane ofP. aeruginosa; tazobactam inhibits β-lactamase allowing the antibacterial activity of piperacillin. In conclusion our data suggest that PEI can be expected to act as a vehicle for experimental drug delivery into cells, and as a potentiating agent for antibacterial agents that are normally excluded by Gram-negative bacteria.     

Antimicrobial resistance of Escherichia coli isolated from freshwaters and hospital effluents in Belgium

The purpose of this work was to evaluate the level of antimicrobial resistant Escherichia coli isolates in freshwaters and hospital effluents in Belgium. The samples were collected from 24 locations along the Ourthe, Vesdre, Amblève and Meuse rivers and in the wastewater effluents of several hospitals. The sampling stations in rivers were classified according to the dominant land covers of the rivers (rural, urban and forest areas). Two sampling campaigns were organized in May and October 2019 to highlight a possible seasonal effect. A total of 938 E. coli strains were isolated on Chromogenic Selective Tryptone Bile X-glucuronide (TBX) and TBX supplemented with amoxicillin (TBX+AMX) media. Disk diffusion assays were performed following the EUCAST's recommendations to assess the antimicrobial resistance against 12 antibiotics. A total of 32·7% of strains were at least resistant to one antibiotic and 24·6% were multiple antimicrobial resistant strains on TBX. The highest resistance rates were found for ampicillin (AMP), amoxicillin coupled with clavulanic acid (AMC) and sulfamethoxazole/trimethoprim (SXT). The lowest resistance rates were observed for meropenem (MEM) and ertapenem (ETP), which are last resort antibiotics. No significant difference was observed between both campaigns for the resistance rate to antibiotics.     

Multiple antibiotic resistances of <Emphasis Type="Italic">Enterococcus</Emphasis> isolates from raw or sand-filtered sewage

Fifty antibiotic-resistant Enterococcus strains were isolated from raw sewage of a wastewater treatment plant and from the same sewage after trickling through a 25-cm sand column, which retained >99% of the initial population. All 50 Enterococcus isolates were resistant against triple sulfa and trimethoprim/sulfamethoxazole and none were resistant against vancomycin. Most of the isolates from raw sewage were resistant to more antibiotics than the isolates from sand column effluent. One Enterococcus isolate from raw sewage (no. 61) and one Enterococcus isolate from sand column effluent (no. 95) had ten antibiotic resistances each. Isolate no. 95 maintained its resistances in the absence of antibiotics during the whole study. It was compared with isolate no. 70, which was one of the isolates, being resistant only against the two sulfonamides. Phenotypically and biochemically, the two organisms were strains of Enterococcus faecalis. Sequence analysis of partical 16S rDNA allowed alignment of isolate no. 95 as a strain of Enterococcus faecium and of isolate no. 70 as a strain of E. faecalis. E. faecium strain no. 95 carried at least six different plasmids, whereas for E. faecalis strain no. 70, no discrete plasmid band was seen on the gels.     

血培养病原菌构成及耐药性分析

了解2008年至2012年哈尔滨医科大学附属第一医院血培养常见病原菌构成及耐药性。对血培养分离出的病原菌进行鉴定,用MIC法、K-B法测定药物敏感性,用WHONET 5.6统计软件进行细菌菌谱及耐药性分析。共分离出病原菌4 245株;其中凝固酶阴性葡萄球菌最多,947株占22.3%;其次为大肠埃希菌822株,肺炎克雷伯菌520株,鲍曼不动杆菌195株,金黄色葡萄球菌142株;耐甲氧西林凝固酶阴性葡萄球菌(MRCNS)和耐甲氧西林金黄色葡萄球菌(MRSA)检出率分别为81.1%、38.8%,未发现耐万古霉素、利奈唑胺及替考拉宁的凝固酶阴性葡萄球菌和金黄色葡萄球菌;大肠埃希菌及肺炎克雷伯菌产ESBLs检出率分别为55.2%、53.8%,对头孢哌酮/舒巴坦、哌拉西林/他唑巴坦、头孢替坦、头孢西丁、阿米卡星的耐药率较低,对碳青霉烯类抗菌药物存在耐药现象;鲍曼不动杆菌对抗菌药物的耐药率普遍较高。及时、准确地对血培养分离出的病原菌进行监测,以便指导临床合理用药,控制耐药株的产生。     

Antimicrobial Susceptibility of Clostridium difficile from Different Sources

A total of 79 Clostridium difficile strains from healthy young and elderly adults, elderly patients without gastrointestinal disease, elderly patients receiving antibiotics without gastrointestinal complications, and elderly patients with antibiotic-associated diarrhea or pseudomembranous colitis were tested for their susceptibilities to 24 antimicrobial agents. All of the 79 strains were inhibited by low concentrations of rifampicin, metronidazole, fusidic acid, vancomycin, ampicillin, and penicillin G. The strains were highly resistant to aminoglycosides, trimethoprim, sulfamethoxazole, nalidixic acid, and cycloserine and often resistant to neomycin, cefoxitin, and cefalexin. Wide variations in the susceptibility of C. difficile strains to erythromycin, clindamycin, lincomycin, chloramphenicol, and tetracycline were found. Strains resistant to erythromycin, clindamycin, and lincomycin were more frequently found among strains isolated from elderly adults than those isolated from young adults, with particularly high frequency among strains isolated from elderly patients receiving antibiotics. None of the 23 strains isolated from healthy young adults was resistant to chloramphenicol. All of the 14 strains resistant to erythromycin, clindamycin, lincomycin, and chloramphenicol were sensitive to tetracycline and all of the 15 strains resistant to erythromycin, clindamycin, lincomycin, and tetracycline were sensitive to chloramphenicol. Only one out of 19 tetracycline-resistant strains was highly toxigenic, whereas 42 (70%) of 60 sensitive strains were highly toxigenic.     

A set of novel multiplex Taqman real-time PCRs for the detection of diarrhoeagenic Escherichia coli and its use in determining the prevalence of EPEC and EAEC in a university hospital

Background

Escherichia coli isolates of equine faecal origin were investigated for antibiotic resistance, resistance genes and their ability to perform horizontal transfer.

Methods

In total, 264 faecal samples were collected from 138 horses in hospital and community livery premises in northwest England, yielding 296 resistant E. coli isolates. Isolates were tested for susceptibility to antimicrobial drugs by disc diffusion and agar dilution methods in order to determine minimum inhibitory concentrations (MIC). PCR amplification was used to detect genes conferring resistance to: ampicillin (TEM and SHV beta-lactamase), chloramphenicol (catI, catII, catIII and cml), tetracycline (tetA, tetB, tetC, tetD, tet E and tetG), and trimethoprim (dfrA1, dfrA9, dfrA12, dfrA13, dfr7, and dfr17).

Results

The proportion of antibiotic resistant isolates, and multidrug resistant isolates (MDR) was significantly higher in hospital samples compared to livery samples (MDR: 48% of hospital isolates; 12% of livery isolates, p < 0.001). Resistance to ciprofloxacin and florfenicol were identified mostly within the MDR phenotypes. Resistance genes included dfr, TEM beta-lactamase, tet and cat, conferring resistance to trimethoprim, ampicillin, tetracycline and chloramphenicol, respectively. Within each antimicrobial resistance group, these genes occurred at frequencies of 93% (260/279), 91%, 86.8% and 73.5%, respectively; with 115/296 (38.8%) found to be MDR isolates. Conjugation experiments were performed on selected isolates and MDR phenotypes were readily transferred.

Conclusions

Our findings demonstrate that E. coli of equine faecal origin are commonly resistant to antibiotics used in human and veterinary medicine. Furthermore, our results suggest that most antibiotic resistance observed in equine E. coli is encoded by well-known and well-characterized resistant genes common to E. coli from man and domestic animals. These data support the ongoing concern about antimicrobial resistance, MDR, antimicrobial use in veterinary medicine and the zoonotic risk that horses could potentially pose to public health.     

Exploring the effectiveness of tazobactam against ceftazidime resistant Escherichia coli: insights from the comparison between susceptibility testing and beta-lactamase inhibition

Thirteen clinical isolates of Escherichia coli resistant to ceftazidime that possessed an AmpC and other (beta-lactamases were identified. The effectiveness of different formulations of piperacillin/tazobactam to other beta-lactams was compared. Antibiotic susceptibility testing, polymerase chain reaction, amplification of blaTEM, blaSHV and blaAmpC, and enzyme-linked immunosorbent assays to identify AmpC beta-lactamases were performed. Hydrolysis rates were obtained and residual enzymatic activity was determined. Cefepime and ertapenem were more active than piperacillin/tazobactam. In contrast, increasing the relative proportion of tazobactam improved susceptibility testing. Twenty micromolar tazobactam inhibited total beta-lactamase activity (as measured by nitrocefin hydrolysis rates) by greater than 75% against all isolates tested: in 11 of 13 E. coli isolates, total beta-lactamase activity was inhibited by 90%. The observed differences between MIC determinations and susceptibility to enzymatic inactivation by tazobactam against E. coli containing AmpC and other -lactamases may be due to the final tazobactam concentration achieved in the periplasmic space. Factors determining this are critical considerations in assessing beta-lactamase inhibitor potency.     

Toxin Production and Antibiotic Resistances in Escherichia coli Isolated from Bathing Areas Along the Coastline of the Oslo Fjord

The presence of enterovirulent and/or antibiotic resistant strains of Escherichia coli in recreational bathing waters would represent a clear health issue. In total, 144 E. coli isolated from 26 beaches along the inner Oslo fjord were examined for virulence determinants and resistance to clinically important antibiotics. No isolates possessed the genetic determinants associated with enterotoxigenic strains and none showed the prototypic sorbitol negative, O157:H7 phenotype. A small number (～1 %) produced alpha-hemolysin. Occurrences and patterns of antibiotic resistances were similar to those of E. coli isolated previously from environmental samples. In total, 6 % of the strains showed one or more clinically relevant resistances and 1.4 % were multi-drug resistant. Microarray analyses suggested that the resistance determinants were generally associated with mobile genetic elements. Resistant strains were not clonally related, and were, furthermore not concentrated at one or a few beach sites. This suggests that these strains are entering the waters at a low rate but in a widespread manner. The study demonstrates that resistant E. coli are present in coastal bathing waters where they can come into contact with bathers, and that the resistance determinants are potentially transferable. Some of the resistances registered in the study are to important antibiotics used in human medicine such as fluoroquinolones. The spread of antibiotic resistant genes, from the clinical setting to the environment, has clear implications with respect to the current management of bacterial infections and the long term value of antimicrobial therapy. The present study is the first of its kind in Norway.     

In vitro Activity of Moxifloxacin Against 179 Strains of Anaerobic Bacteria Found in Pulmonary Infections

The activity of moxifloxacin (BAY 12-8039), a new 8-methoxyquinolone, was determined using the NCCLS-approved Wadsworth brucella laked blood agar method and compared to the activities of metronidazole, penicillin G, piperacillin/tazobactam and trovafloxacin. Breakpoints used to define susceptible and resistant categories were, respectively: ≤ 8 and ≥ 32 μg/mL for metronidazole, ≤ 2 and ≥ 8 μg/mL for moxifloxacin and trovafloxacin, ≤ 0.5 and ≥ 2 μg/mL for penicillin G and ≤ 32 and ≥128 μg/mL for piperacillin/tazobactam. A total of 179 anaerobic isolates from pulmonary infections were tested. Piperacillin/tazobactam was the most active antimicrobial, inhibiting 99% of strains at the susceptible breakpoint. Ninety-seven percent of these isolates were susceptible to moxifloxacin; 96% to trovafloxacin, 89% to metronidazole and 43% to penicillin G. Geometric mean moxifloxacin MIC values forBacteroides fragilis and the B. fragilis group were 0.5 and 0.8 μg/mL, respectively. Eighty-eight percent of B. fragilis and 100% of other B. fragilis group species were susceptible to both moxifloxacin and trovafloxacin. All of the strains of B. fragilis and most of the other B. fragilis group species were resistant to penicillin G. At least 99% of other Bacteroides species, Prevotella, and Fusobacterium strains were susceptible to moxifloxacin, metronidazole, piperacillin/tazobactam and trovafloxacin (88% were susceptible to trovafloxacin at 2 μg/mL and all were susceptible at 4 μg/mL). The strains of Clostridium difficile andClostridium ramosum found in these specimens were both resistant to penicillin G but susceptible to the other agents. All strains of Peptostreptococcus species were susceptible to all of the agents except penicillin G. Activities of the agents against non-spore-forming Gram-positive rods at the intermediate breakpoint were, respectively, moxifloxacin-100%, metronidazole-49%, penicillin G-86%, piperacillin/tazobactam-100%, and trovafloxacin-97%. The promising in vitro activity of moxifloxacin against anaerobic pulmonary isolates warrants further investigation, including clinical correlation studies.     

Efficacy of Mastoparan-AF alone and in combination with clinically used antibiotics on nosocomial multidrug-resistant Acinetobacter baumannii

Emergence of multidrug-resistant Acinetobacter baumannii (MDRAB) has become a critical clinical problem worldwide and limited therapeutic options for infectious diseases caused by MDRAB. Therefore, there is an urgent need for the development of new antimicrobial agents or alternative therapy to combat MDRAB infection. The aim of this study was to investigate effects of Mastoparan-AF (MP-AF), an amphipathic peptide isolated from the hornet venom of Vespa affinis with broad-spectrum antimicrobial activity, on MDRAB. As compared with clinical used antibiotics, MP-AF exhibited potent antimicrobial activity at 2–16 μg/ml against the reference strain A. baumannii ATCC 15151 and seven MDRAB clinical isolates, especially the colistin-resistant MDRAB, E0158. The synergistic antimicrobial combination study revealed that MP-AF acted synergistically with specific antibiotics, e.g., ciprofloxacin, trimethoprim/sulfamethoxazole (SXT) or colistin against some isolates of the MDRAB. It was noteworthy when MP-AF combined with SXT exhibited synergistic activity against all SXT-resistant MDRAB isolates. The synergistic combination of MP-AF and antibiotics could reduce the dosage recommended of each antimicrobial agent and improve the safety of medications with ignorable adverse effects, such as colistin with nephrotoxicity in therapeutic dose. Furthermore, MP-AF combined with antibiotics with different antimicrobial mechanisms could reduce selective pressure of antibiotics on bacteria and prevent the emergence of antimicrobial-resistant strains. Importantly, we are the first finding that MP-AF could make MDRAB from the original non-susceptibility to SXT become sensitivity. In conclusion, MP-AF alone or in combination with other antibiotics, especially SXT, is a potential candidate against MDRAB infection in clinical medicine.     

Phenotypic and Molecular Characterization of Multidrug Resistant Klebsiella pneumoniae Isolated from a University Teaching Hospital,China

The multidrug-resistant rate of Klebsiella pneumoniae has risen rapidly worldwide. To better understand the multidrug resistance situation and molecular characterization of Klebsiella pneumoniae, a total of 153 Klebsiella pneumoniae isolates were collected, and drug susceptibility test was performed to detect its susceptibility patterns to 13 kinds of antibiotics. Phenotypic tests for carbapenemases ESBLs and AmpC enzyme-producing strains were performed to detect the resistance phenotype of the isolates. Then PCR amplification and sequencing analysis were performed for the drug resistance determinants. The results showed that 63 strains harbored bla _CTX-M gene, and 14 strains harbored bla _DHA gene. Moreover, there were 5 strains carrying bla _KPC gene, among which 4 strains carried bla _CTX-M, bla _DHA and bla _KPC genes, and these 4 strains were also resistant to imipenem. Our data indicated that drug-resistant Klebsiella pneumoniae were highly prevalent in the hospital. Thus it is warranted that surveillance of epidemiology of those resistant isolates should be a cause for concern, and appropriate drugs should be chosen.