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In vitro antimicrobial activity of moxifloxacin compared to other quinolones against recent clinical bacterial isolates from hospitalized and community-based cancer patients.

The in vitro spectrum of moxifloxacin (a C-8-methoxyquinolone) was compared to that of ciprofloxacin and levofloxacin against 924 recent clinical isolates from cancer patients. Moxifloxacin was more active than the comparator agents against Gram-positive pathogens, with potent activity against Aerococcus spp., Listeria monocytogenes, Micrococcus spp., Rhodococcus equi, and Stomatococcus mucilaginous, methicillin-susceptible Staphylococcus spp., all beta hemolytic streptococci, viridans streptococci and Streptococcus pneumoniae. It also had good to moderate activity against Bacillus spp., Corynebacterium spp., Enterococcus faecalis, and methicillin-resistant staphylococci. Although ciprofloxacin was the most active agent tested against the Enterobacteriaceae, moxifloxacin inhibited the majority of these isolates at < or =2.0 microg/ml. Moxifloxacin was the least active of the three agents tested against Pseudomonas aeruginosa, but had significant activity against other non-fermentative Gram-negative bacilli including Acinetobacter spp., Flavobacterium spp., Pseudomonas spp. other than P. aeruginosa, and Stenotrophomonas maltophilia. The overall broad spectrum of moxifloxacin, and its availability for both oral and parenteral administration, warrants its evaluation for the prevention and treatment of infections in cancer patients.

Anti-Infective Agents↗

Frequency, epidemiology, risk factors, clinical and bacteriological features of enterococcal disease in patients with HIV infection in a decade survey.

In order to assess the frequency and clinical significance of Enterococcus spp. infection during HIV disease, the epidemiological features, risk factors, microbiological issues, and therapeutic perspectives of all the 148 consecutive episodes observed in the past decade were analyzed. The overall frequency of these complications (which involved the genito-urinary tract in over 75% of cases) regarded 5.3% of all admission for HIV disease, with a clear prevalence of Enterococcus faecalis as the causative agent (86.5% of episodes), and a proportionally elevated frequency of polymicrobial infection (45.9% of cases). Among the 148 cultured bacterial strains, a complete susceptibility to glycopeptide antibiotics was documented, together with favorable sensitivity levels against semisynthetic penicillins, followed by chloramphenicol, macrolides, and clindamycin. An advanced underlying HIV disease characterized by a concurrent, severe immunodeficiency, concomitant, prolonged neurological complications, hospitalization itself (with prevalence of nosocomial infection), recourse to invasive diagnostic and/or therapeutic procedures, and prior administration of broad spectrum antimicrobial agents, all seem to support HIV-associated enterococcal disease (mostly involving the genito-urinary tract). The adjunct of neutropenia, a very low CD4+ lymphocyte count, and severe AIDS-defining illnesses represented significant risk factors for hematogenous dissemination of this bacterial infection with potentially life-threatening consequences (16.1% of lethal cases among our septic patients).

AIDS-Related Opportunistic Infections↗

Chemiluminescence of enterococci isolates from freshwater.

All Enterococcus spp., isolated from environmental water samples (n=81), emitted a high chemiluminescence signal in the presence of luminol (10(-2) M). Kinetic studies of chemiluminescence show a close correlation between chemiluminescence and growth curves during the exponential phase, with a maximum chemiluminescence reached just before bacterial growth entered in the stationary phase. On the other hand, genera closely related to Enterococcus such as Streptococcus or Lactococcus produced a very weak chemiluminescent signal. Chemiluminescence of enterococci could therefore offer a rapid test, in aiding the identification of the genus Enterococcus and in the survey of the microbiological quality of water supplies.

Bacterial Typing Techniques↗

Impact of integrated fish farming on antimicrobial resistance in a pond environment.

Integrated fish farming combines livestock production with fish farming. Animal manure is shed directly into a fish pond as fertilizer and supports the growth of photosynthetic organisms. The livestock, mainly chickens and pigs, is often fed feed containing growth promoters. In this study we investigated the impact of integrated fish farming on the levels of antimicrobial-resistant bacteria in a pond environment. One integrated broiler chicken-fish farm was studied for 2 months immediately after the start of a new fish production cycle. A significant increase over time in the resistance to six different antimicrobials was found for the indicator organism Acinetobacter spp. isolated from composite water-sediment samples. The initial resistance levels prior to the new production cycle were 1 to 5%. After 2 months the levels of resistance to oxytetracycline and sulfamethoxazole reached 100%, and the levels of resistance to ciprofloxacin were more than 80%. The long-term effects of resistance on integrated farming were studied on seven additional farms. The resistance levels were particularly high among Enterococcus spp. and were also high among Acinetobacter spp. isolated from water-sediment samples compared to the resistance levels at four control farms. In conclusion, integrated fish farming seems to favor antimicrobial-resistant bacteria in the pond environment. This could be attributed to the selective pressure of antimicrobials in the pond environment and/or to the introduction of antimicrobial-resistant bacteria from animal manure. Potential risks to human health were not addressed in this study and remain to be elucidated.

Acinetobacter↗

Antimicrobial resistance of Enterococcus species isolated from produce.

The purpose of this study was to characterize the antibiotic resistance profiles of Enterococcus species isolated from fresh produce harvested in the southwestern United States. Among the 185 Enterococcus isolates obtained, 97 (52%) were Enterococcus faecium, 38 (21%) were Enterococcus faecalis, and 50 (27%) were other Enterococcus species. Of human clinical importance, E. faecium strains had a much higher prevalence of resistance to ciprofloxacin, tetracycline, and nitrofurantoin than E. faecalis. E. faecalis strains had a low prevalence of resistance to antibiotics used to treat E. faecalis infections of both clinical and of agricultural relevance, excluding its intrinsic resistance patterns. Thirty-four percent of the isolates had multiple-drug-resistance patterns, excluding intrinsic resistance. Data on the prevalence and types of antibiotic resistance in Enterococcus species isolated from fresh produce may be used to describe baseline antibiotic susceptibility profiles associated with Enterococcus spp. isolated from the environment. The data collected may also help elucidate the role of foods in the transmission of antibiotic-resistant strains to human populations.

Anti-Bacterial Agents↗

Proficiency of clinical laboratories in and near Monterrey, Mexico, to detect vancomycin-resistant enterococci.

Early detection of vancomycin-resistant enterococci is important for preventing its spread among hospitalized patients. We surveyed the ability of eight hospital laboratories in and near Monterrey, Mexico, to detect vancomycin resistance in Enterococcus spp. and found that although laboratories can reliably detect high-level vancomycin resistance, many have difficulty detecting low-level resistance.

Anti-Bacterial Agents↗

Development of a novel amplified fragment length polymorphism (AFLP) typing method for enterococci isolates from cattle faeces and evaluation of the single versus pooled faecal sampling approach.

A novel and highly reproducible amplified fragment length polymorphism (AFLP) typing approach was developed for typing of Enterococcus strains from the environment. Pooled and corresponding single faecal sample isolates were analysed to test the efficiency and coverage of dominant isolates for future sampling procedures. AFLP development was based on the selection of appropriate restriction enzymes and the design of adaptors and primers which was supported by in silico optimisation of selective bases using Enterococcus spp. genome data. Three optimal combinations of selective bases at the 3' end of the designed primers (i.e., CC, GG, CG) could be determined. AFLP fragment analysis using a capillary sequencer and intralane standardisation resulted in excellent methodical stability (> or =98% similarity for GG and > or =94% similarity for CC). Furthermore, the developed typing method was evaluated on 16 type trains of the genera Enterococcus and Streptococcus and 398 faecal isolates of cow pats from five alpine pastures in a karstic catchment area. Statistical analysis revealed a discrimination capacity of DI > or =0.95 (Simpson Diversity Index) and a reproducibility level of > or =94% similarity indicating the methods high typing capacity and robustness. Results of the comparative analysis of single and pooled faecal samples indicate that for a "strain to strain" based faecal source tracking, pooled faecal samples rather than single faecal samples are likely to be the most efficient sampling strategy for collecting the abundant corresponding strains.

Animals↗

In vitro activities of DX-619 and comparison quinolones against gram-positive cocci.

The in vitro activity of the novel quinolone DX-619 was compared to those of currently available quinolones against U.S. clinical isolates of Staphylococcus aureus, coagulase-negative staphylococci, Enterococcus spp., Streptococcus pyogenes, and Streptococcus pneumoniae. DX-619 was the most potent quinolone overall, indicating possible utility as an anti-gram-positive quinolone.

Anti-Infective Agents↗

[Year to year changes in isolation frequencies of different bacteria from postoperative infections].

Bacterial species isolated from postoperative infections during the period from July 1982 to June 1993 were studied regarding isolation frequencies of different species in different years in a joint research project involving nine (9) university hospitals in Japan. The obtained results are summarized as follows. (1) Altogether, 1,453 strains were isolated with 153 strains obtained during the most recent one year from July of 1992 through June of 1993. The most numerous source of isolation was postoperative intra-abdominal sepsis. (2) Gram-positive cocci were the most often isolated, among them methicillin-resistant Staphylococcus aureus was isolated in increasing frequencies until June, 1992. (3) In the last one year, Enterococcus spp. was isolated the most from postoperative infections followed by Staphylococcus spp. and Pseudomonas spp. Rates of isolation of anaerobic bacteria were relatively low with annual isolation frequencies ranging 8 to 15 per cent.

Digestive System Diseases↗

Microbiological evaluation of bottled non-carbonated ("still") water from domestic brands in Greece.

The microbiological quality of 1,527 samples of bottled non-carbonated ('still') mineral water, purchased from retail outlets and derived from 10 manufacturing companies in Greece, was investigated during the period 1995-2003. Applying the membrane filter technique, the aliquots of water samples (250 ml) were analyzed for the presence and enumeration of total coliforms, Escherichia coli, Enterococcus spp. and Pseudomonas aeruginosa. Also, aerobic bacteria were counted as Heterotrophic Plate Count (HPC) ml(-1) at 22 and 37 degrees C. Positive samples for the parameters tested varied significantly among brands with an overall percentage of 13.95% bottled water samples noncompliant with the Greek water regulation. Microorganisms isolated from the samples tested were identified as species of Pseudomonas, Aeromonas, Pasteurella, Citrobacter, Flavobacterium, Providencia and Enterococcus. The most frequent isolated microorganism during the period of the study was P. aeruginosa. Generally, bacterial load of the samples tested ranged in low levels. The purpose of the current study was to evaluate the microbiological quality of the bottled water provided by domestic brands in the Greek market during the period 1995-2003.

Bacteria↗

Influence of transferable genetic determinants on the outcome of typing methods commonly used for Enterococcus faecium.

A variety of methods is used for a molecular typing of Enterococcus spp. and related gram-positive bacteria including macrorestriction analysis using pulsed-field gel electrophoresis (PFGE), ribotyping, rapid amplification of polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP). To test the influence of transferable determinants on the outcome of different typing methods commonly used for enterococci, we established a homogenous strain collection of 24 transconjugants resulting from filter matings with antibiotic-resistant Enterococcus faecium. As expected, AFLP, RAPD, and PFGE all identified our model bacteria as strongly related. However, distinct differences in the resolving and discriminatory power of the tested methods could be clearly addressed. In PFGE, 22 of 24 transconjugants possessed less than a three-band difference to the recipient pattern and would be regarded as strongly related. Three different RAPD PCRs were tested; in two reactions, identical patterns for all transconjugants and the recipient were produced. One RAPD PCR produced an identical pattern for 18 transconjugants and the recipient and a clearly different pattern for the remaining 6 transconjugants due to a newly appearing fragment resulting from acquisition of the tetL gene. AFLP clusters all transconjugants into a group of major relatedness. Percent similarities were highly dependent on the method used for calculating the similarity coefficient (curve-based versus band-based similarity coefficient). Fragment patterns of digested plasmids showed the possession of nonidentical plasmids in most transconjugants. PFGE still could be recommended as the method of choice. Nevertheless, the more-modern AFLP approach produces patterns of comparable discriminatory power while possessing some advantages over PFGE (less-time-consuming internal standards). Plasmid fingerprints can be included to subdifferentiate enterococcal isolates possessing identical macrorestriction and PCR typing patterns.

Anti-Bacterial Agents↗

Spectrum of aetiological agents of postoperative endophthalmitis and antibiotic susceptibility of bacterial isolates.

PURPOSE: To determine the spectrum of infectious agents of postoperative endophthalmitis, the relationship with the time of onset of symptoms after surgery and the antibiotic susceptibilities of the aerobic bacterial isolates. METHODS: A retrospective review of microbiological records from January 1995 to December 1998 yielded 173 isolates from intraocular specimen of 170 patients with culture-proven postoperative endophthalmitis. Antibiotic susceptibility of these isolates was determined for various ocular antibiotics using the Kirby-Bauer disk-diffusion test. Based on the time of onset of illness, clinical presentation was classified into acute, delayed and chronic. RESULTS: Among 170 cases, 71 (41.7%) were attributable to gram-negative, 64 (37.6%) to gram-positive bacteria, and 37 (21.8%) to fungi. Gram-negative bacteria included P. aeruginosa (29;17.1%), other Pseudomonas spp (15;8.8%), non-fermenters (18;10.6%) and others (10;5.8%). Among these, 40 of 72 (55.5%) were sensitive to gentamicin, 47 of 72 (65.2%) to cefotaxime, 47 of 69 (68.1%) to amikacin, 52 of 71 (73.2%) to ciprofloxacin, and 25 of 40 (62.5%) to ceftazidime. The gram-positive bacteria included S. epidermidis (22;12.9%), S. aureus (13;7.6%), P. acnes (10;5.9%), Enterococcus spp (4;2.3%), Streptococcus spp (7;4.1%) and others (8;4.8%). Among these, 41 of 53 (77.3%) were sensitive to gentamicin, 47 of 53 (88.6%) to cefotaxime, 46 of 52 (88.4%) to ciprofloxacin, 38 of 41 (92.6%) to cefazolin and 27 of 37 (72.9%) to ceftazidime. All gram-positive bacteria were sensitive to vancomycin. CONCLUSION: In this large series of postoperative endophthalmitis, gram-negative bacilli followed by fungi accounted for the largest number of cases. A high degree of resistance of gram-negative bacilli to gentamicin, cefotaxime, amikacin and ceftazidime was recorded.

Anti-Bacterial Agents↗

Enterococcus gilvus sp. nov. and Enterococcus pallens sp. nov. isolated from human clinical specimens.

Light yellow-pigmented (strain PQ1) and yellow-pigmented (strain PQ2), gram-positive, non-spore-forming, nonmotile bacteria consisting of pairs or chains of cocci were isolated from the bile of a patient with cholecystitis (PQ1) and the peritoneal dialysate of another patient with peritonitis (PQ2). Morphologically and biochemically, the organisms phenotypically belonged to the genus Eterococcus. Whole-cell protein (WCP) analysis and sequence analysis of a segment of the 16S rRNA gene suggested that they are new species within the genus Enterococcus. PQ1 and PQ2 displayed less than 70% identities to other enterococcal species by WCP analysis. Sequence analysis showed that PQ1 shared the highest level of sequence similarity with Enterococcus raffinosus and E. malodoratus (sequence similarities of 99.8% to these two species). Sequence analysis of PQ2 showed that it had the highest degrees of sequence identity with the group I enterococci E. malodoratus (98.7%), E. raffinosus (98.6%), E. avium (98.6%), and E. pseudoavium (98.6%). PQ1 and PQ2 can be differentiated from the other Enterococcus spp. in groups II, III, IV, and V by their phenotypic characteristics: PQ1 and PQ2 produce acid from mannitol and sorbose and do not hydrolyze arginine, placing them in group I. The yellow pigmentation differentiates these strains from the other group I enterococci. PQ1 and PQ2 can be differentiated from each other since PQ1 does not produce acid from arabinose, whereas PQ2 does. Also, PQ1 is Enterococcus Accuprobe assay positive and pyrrolidonyl-beta-naphthylamide hydrolysis positive, whereas PQ2 is negative by these assays. The name Enterococcus gilvus sp. nov. is proposed for strain PQ1, and the name Enterococcus pallens sp. nov. is proposed for strain PQ2. Type strains have been deposited in culture collections as E. gilvus ATCC BAA-350 (CCUG 45553) and E. pallens ATCC BAA-351 (CCUG 45554).

Bacterial Proteins↗

[Cryptosporidium y Giardia in surface water].

The aim of this work was to determine Giardia and Cryptosporidium levels in surface water used either as recreational or drinking water source, and to estimate the relationship with bacteriological and physicochemical parameters determining water quality. This study was carried out during 22 months, in Santa Fe and Santo Tomé cities (Santa Fe Province, Argentina). Cryptosporidium from 100% of the recreational samples (44-2404 oocysts/100 l) and from 92% of water sources (< 20-539 oocysts/100 l) was detected, while Giardia was found in lower concentrations and frequencies: 72% (< 20-670 cysts/100 l) and 31% (< 20-65 cysts/100 l), respectively. Statistically significant relationships were found between these parasites and organic matter, turbidity and bacteriological indicators of contamination (Escherichia coli, streptococci, Enterococcus spp, Pseudomonas aeruginosa). These relationships, however, vary according to the sampling place, the source and levels of contamination.

Animals↗

Evaluation of antibiotic resistance analysis and ribotyping for identification of faecal pollution sources in an urban watershed.

AIMS: The accuracy of ribotyping and antibiotic resistance analysis (ARA) for prediction of sources of faecal bacterial pollution in an urban southern California watershed was determined using blinded proficiency samples. METHODS AND RESULTS: Antibiotic resistance patterns and HindIII ribotypes of Escherichia coli (n = 997), and antibiotic resistance patterns of Enterococcus spp. (n = 3657) were used to construct libraries from sewage samples and from faeces of seagulls, dogs, cats, horses and humans within the watershed. The three libraries were analysed to determine the accuracy of host source prediction. The internal accuracy of the libraries (average rate of correct classification, ARCC) with six source categories was 44% for E. coli ARA, 69% for E. coli ribotyping and 48% for Enterococcus ARA. Each library's predictive ability towards isolates that were not part of the library was determined using a blinded proficiency panel of 97 E. coli and 99 Enterococcus isolates. Twenty-eight per cent (by ARA) and 27% (by ribotyping) of the E. coli proficiency isolates were assigned to the correct source category. Sixteen per cent were assigned to the same source category by both methods, and 6% were assigned to the correct category. Addition of 2480 E. coli isolates to the ARA library did not improve the ARCC or proficiency accuracy. In contrast, 45% of Enterococcus proficiency isolates were correctly identified by ARA. CONCLUSIONS: None of the methods performed well enough on the proficiency panel to be judged ready for application to environmental samples. SIGNIFICANCE AND IMPACT OF THE STUDY: Most microbial source tracking (MST) studies published have demonstrated library accuracy solely by the internal ARCC measurement. Low rates of correct classification for E. coli proficiency isolates compared with the ARCCs of the libraries indicate that testing of bacteria from samples that are not represented in the library, such as blinded proficiency samples, is necessary to accurately measure predictive ability. The library-based MST methods used in this study may not be suited for determination of the source(s) of faecal pollution in large, urban watersheds.

Animals↗

Comparison of a rabbit model of bacterial endocarditis and an in vitro infection model with simulated endocardial vegetations.

Animal models are commonly used to determine the efficacy of various antimicrobial agents for treatment of bacterial endocarditis. Previously we have utilized an in vitro infection model, which incorporates simulated endocardial vegetations (SEVs) to evaluate the pharmacodynamics of various antibiotics. In the present study, we compared four experimental rabbit endocarditis protocols to an in vitro infection model in an effort to determine if these models are comparable. We have evaluated the activity of clinafloxacin, trovafloxacin, sparfloxacin, and ciprofloxacin in rabbit models against Staphylococcus aureus and Enterococcus spp. In vitro models were performed simulating the antibiotic pharmacokinetics obtained in the in vivo studies. Models were dosed the same as rabbit models, and SEVs were evaluated at the same time the rabbit vegetations were examined. Clinafloxacin and trovafloxacin were evaluated against methicillin-susceptible (MSSA1199) and -resistant (MRSA494) strains of S. aureus. Ciprofloxacin was studied against MSSA1199 and MSSA487. Sparfloxacin and clinafloxacin were evaluated against Enterococcus faecium SF2149 and Enterococcus faecalis WH245, respectively. We found that reductions in SEV bacterial density obtained in the in vitro model were similar to those obtained in rabbit vegetations, indicating that the SEV model may be a valuable tool for assessing antibiotic potential in the treatment of bacterial endocarditis.

Animal Testing Alternatives↗

[Bacteria isolated from surgical infections and its susceptibilities to antimicrobial agents--special references to bacteria isolated between April 2003 and March 2004].

Tendency of isolated bacteria from infections in general surgery during the period from April 2004 to March 2005 were investigated in a multicenter study in Japan, and the following results were obtained. In this series, 645 strains including 17 strains of Candida spp. were isolated from 226 (79.0%) of 286 patients with surgical infections. Three hundred and seventeen strains were isolated from primary infections, and 345 strains were isolated from postoperative infections. From primary infections, anaerobic Gram-positive bacteria and anaerobic Gram-negative bacteria were predominant, while aerobic Gram-positive bacteria were predominant from postoperative infections. The isolation rate of aerobic Gram-positive bacteria, such as Enterococcus spp. and Staphylococcus aureus were higher from both types of infections. Among anaerobic Gram-positive bacteria, the isolation rate of Peptostreptococcus spp. was the highest from both types of infections. Among aerobic Gram-negative bacteria, Escherichia coli was the most predominantly isolated from primary infections, followed by Pseudomonas aeruginosa, Klebsiella pneumoniae and Citrobacter freundii in this order, and from postoperative infections, P. aeruginosa was the most predominantly isolated, followed by E. coli, E. cloacae, and K. pneumoniae. Among anaerobic Gram-negative bacteria, the isolation rate of Bacteroides fragilis group was the highest from both primary infections followed by Bilophila wadsworthia. While the isolation rate of B. fragilis group was also the highest from postoperative infections, the following bacteria were Bacteroides thetaiotaomicron and B. wadsworthia in this order. In this series, we noticed no vancomycin-resistant Gram-positive cocci, but a few strains of moderately arbekacin-resistant MRSA. Carbapenem-resistant P. aeruginosa but not multidrug-resistant was seen in 13.3 per cents. Also cefazolin-resistant E. coli probably producing extended spectrum beta-lactamase was seen in 7.0 per cents. We should be carefully followed up the facts that an increasing isolation rates of B. fragilis group and B. wadsworthia which were resistant to both penicillins and cephems.

Anti-Bacterial Agents↗

Antibacterial activity of cefpodoxime in vitro.

Cefpodoxime proxetil is a new orally administered prodrug which is absorbed and de-esterified by the intestinal mucosa to release the third-generation cephalosporin, cefpodoxime, and which is undergoing in vitro and in vivo evaluations. Using the standard agar dilution method, we compared the in vitro activity of this drug with other oral cephalosporins and quinolones against 637 recent clinical isolates from Kaohsiung Veterans General Hospital in Taiwan. Against Escherichia coli and Klebsiella pneumoniae, cefpodoxime showed excellent activity, inhibiting over 90% of these isolates at 1 mg/l. Like other oral drugs of its class, it had little activity against Pseudomonas aeruginosa and Acinetobacter spp. Against Haemophilus influenzae, irrespective of beta-lactamase production, its activity was similar to comparative drugs. Against methicillin-susceptible Staphylococcus aureus, cefpodoxime showed moderate activity, inhibiting 90% of these isolates at 4 mg/l, whereas it was inactive against methicillin-resistant S. aureus. However, all cephalosporins have shown little in vivo activity against methicillin-resistant S. aureus regardless of in vitro results. Cefpodoxime was inactive against Enterococcus spp. Against other streptococci, its activity was similar to other oral cephalosporins and quinolones tested. The results of this in vitro study indicated that oral administration of cefpodoxime should be an ideal agent in the empirical outpatient treatment for community-acquired cutaneous, respiratory and urinary tract infections.

Anti-Bacterial Agents↗