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PubMed2006

Antimicrobial resistance among faecal enterococci from healthy individuals in Portugal.

Analysis of 247 faecal enterococcal isolates from 99 healthy Portuguese individuals during 2001 revealed the presence of enterococci resistant to vancomycin (5%) and highly resistant to streptomycin (52%), kanamycin (40%) or gentamicin (11%). Most isolates were also resistant to tetracycline, erythromycin, ciprofloxacin and quinupristin-dalfopristin. The vanA (two Tn1546 types), vanC1, erm(B), aac(6')-aph(2'')-Ia, aph(3')-IIIa, vat(E) and vat(D) genes were detected. Enterococcus faecalis and Enterococcus faecium isolates with high-level resistance to gentamicin were related to Portuguese poultry isolates described previously. E. faecium isolates that were highly resistant to vancomycin or gentamicin harboured different housekeeping purK alleles associated previously with different hosts.

Animals↗
PubMed2006

Multiple antibiotic resistances of Enterococcus 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.

Anti-Bacterial Agents↗
PubMed1999

Comparison of simple and rapid methods for identifying enterococci intrinsically resistant to vancomycin.

Three different methodologies, reduction of litmus milk (LM) and acidification of arabinose (ARA), acidification of methyl-alpha-D-glucopyranoside (MGP), and rapid motility (RM), for differentiating isolates of Enterococcus casseliflavus and Enterococcus gallinarum (intrinsically vancomycin-resistant enterococci [IVRE]) from Enterococcus faecalis and Enterococcus faecium were evaluated. All 33 isolates of E. faecalis tested reduced LM within 4 h and were negative in all other tests, while the 53 isolates of E. faecium were ARA positive only. In contrast, 45 of 46 (98%) IVRE isolates examined (26 E. casseliflavus and 20 E. gallinarum isolates) acidified MGP, 41 of 46 (89%) were LM and ARA positive, and 45 of 46 (98%) were RM positive. Acidification of MGP was therefore the single most useful test for differentiating IVRE from vancomycin-resistant E. faecium and E. faecalis; however, a combination of LM-ARA and RM testing enabled the correct designation of organisms without the need for overnight incubation.

Anti-Bacterial Agents↗
PubMed2006

Antimicrobial disinfection effect of a laundering procedure for hospital textiles against various indicator bacteria and fungi using different substrates for simulating human excrements.

Recent studies confirm the increase of nosocomial infections and microbial resistance. One of the possible causes is infected textiles due to inappropriate laundering procedures. Most Slovenian laundries use thermal laundering procedures with high energy and water consumption to disinfect hospital textiles. In addition to this fact, there is an increasing number of hospital textiles composed of cotton/polyester blends that cannot endure high temperatures of thermal disinfection. On the other hand, decreasing the temperature of laundering procedures enhances the possibility of pathogenic microorganisms to survive the laundering procedure. In our research, we determined the antimicrobic laundering effect by simulating a common laundering procedure for hospital textiles in the laboratory washing machine at different temperatures by the use of bioindicators. Enterococcus faecium, Staphylococcus aureus, Mycobacterium terrae, Enterobacter aerogenes, and Pseudomonas aeruginosa were used for determining the antibacterial laundering effect. Candida albicans was used for determining the antifungal laundering effect. Swine blood, artificial sweat, and swine fat were used as substrates for simulating human excrements and were inoculated together with the chosen microorganisms onto cotton pieces to simulate real laundering conditions. It was found that E. faecium, S. aureus, E. aerogenes, and P. aeruginosa survived at 60 degrees C, but no microorganisms were found at 75 degrees C.

Animals↗
PubMed2001

Evaluation of the components of a commercial probiotic in gnotobiotic mice experimentally challenged with Salmonella enterica subsp. enterica ser. Typhimurium.

Vitacanis((R)), a probiotic preparation containing a Lactobacillus acidophilus, an Enterococcus faecium and a Saccharomyces cerevisiae, has been developed for the prevention of intestinal disorders in dogs and cats. In the present study, these microorganisms were tested jointly or singly during experimental infection of gnotobiotic mice with Salmonella Typhimurium. Four experimental groups consisting of animals given probiotics jointly or singly and a control group consisting of germfree mice were used. The groups were treated with one or three of the microorganisms (experimental) or PBS (control) 10 days before intragastric challenge with a suspension containing about 10(2) cells of the bacterial pathogen. A higher survival (P<0.05) was observed in gnotobiotic mice given E. faecium (82%). All the animals in the other groups died after the challenge but the survival time was longer (P<0.05) for groups given all three of the microorganisms (7.4+/-2.4 days) or given only L. acidophilus (7.2+/-2.9 days) than for the control mice (4.4+/-1.1 days) and the mice that received S. cerevisiae (4.9+/-1.6 days) mice. The survival data agreed with the histopathological findings which showed more severe liver and intestinal lesions in control mice and in mice given Saccharomyces. In vitro antagonistic assays showed inhibition growth of E. faecium and S. Typhimurium around the colonies of L. acidophilus and for S. Typhimurium around the colonies of E. faecium. However, in vivo, S. Typhimurium became similarly established in the digestive tract of gnotobiotic mice at levels ranging from 10(8) to 10(10)CFU/g of feces and remained at these high levels until the animals died or were sacrificed. Among the three probiotic components of the commercial product Vitacanis((R)), E. faecium was the only one that provided protection against challenge with S. Typhimurium. Protection was not due to the reduction of the intestinal populations of the pathogenic bacteria.

Animals↗
PubMed2006

Shortcut detection of the vanB gene cluster in enterococci by a duplex real-time PCR assay.

AIM: To develop a robust, simple and rapid method for detection of vanB in enterococci. METHODS: A real-time duplex PCR assay for the simultaneous detection of Enterococcus faecium and vanB resistance genotype in enterococci was developed in conjunction with a simple method for DNA extraction. The assay was tested on 130 fresh plate cultures of clinical isolates of enterococci and other Gram-positive bacteria. RESULTS: Forty-eight isolates of vanB E. faecium from 32 different patients and three isolates of vanB E. faecalis were detected within 1 hour. All isolates of E. faecium were identified correctly. CONCLUSION: This simple method for the detection of resistance mediated by vanB is a potentially useful method that is suitable for use in the diagnostic microbiology laboratory.

Anti-Bacterial Agents↗
PubMed1999

[Successful modification of human intestinal microflora with oral administration of lactic acid bacteria].

Lactic acid bacteria in food can transiently colonize the intestine and exert health beneficial (probiotic) effects. These include: 1. Lactose digestion, improvement of diarrheal disorders (including traveller's diarrhea), prophylaxis of intestinal and urogenital infections--as a result of formation or reconstruction of a balanced indigenous microflora. 2. Inhibition of the mutagenicity of the intestinal contents and reduction of the incidence of intestinal tumours. 3. Immunomodulatory effects resulting in the improved host resistance. 4. Depression of the serum cholesterol level. The most of these effects were observed in a group of adult subjects administered daily by a lyophilized Enterococcus faecium M-74 in the form of waffles (Dr. Ebi) during nine weeks of a double blind placebo controlled clinical trial. The bacterium temporarily colonized the host intestine and its secretion in stool persisted for six weeks after the last dose. The mean activities of beta-D-glucuronidase in stools of subjects given waffles containing enterococci were reduced comparing to stools of placebo subjects. After six weeks of daily eating the waffles with enterococci, an increased production of superoxide and other reactive oxygen intermediates by peripheral neutrophils was observed. The increase corresponded in time with an elevated formation of IgG by peripheral blood mononuclear cells after polyclonal activation with mitogenes. Higher activities of myeloperoxidase and elastase in peripheral neutrophils were also ascertained during eating of waffles containing of E. faecium M-74. Hence, intake of E. faecium M-74 in the form of waffles may have an significant immunostimulatory effect on both phagocytosis performed by neutrophils and antibody production. (Tab. 6, Ref. 29.)

Administration, Oral↗
PubMed1996

Quantitative analysis of the metabolism of soluble cytoplasmic peptidoglycan precursors of glycopeptide-resistant enterococci.

Transposon Tn1546 from Enterococcus faecium BM4147 mediates high-level resistance to the glycopeptide antibiotics vancomycin and teicoplanin. Tn 1546 encodes a dehydrogenase (VanH) and a ligase (VanA) that synthesize D-alanyl-D-lactate (D-Ala-D-Lac), a D,D-dipeptidase (VanX) that hydrolyses D-Ala-D-Ala and a two-component regulatory system (VanR-VanS) that controls transcription of the vanHAX operon. Strains of Enterococcus faecalis harbouring various copy numbers of the vanRSHAX cluster were tested to determine if there was a correlation between the levels of resistance to glycopeptides, the levels of expression of the corresponding resistance genes and the relative proportions of the different cytoplasmic peptidoglycan precursors. Increased transcription of the vanHAX operon was associated with increased incorporation of D-Ala-D-Lac into peptidoglycan precursors to the detriment of D-Ala-D-Ala, and with a gradual increase in the vancomycin-resistance levels. More complete elimination of D-Ala-D-Ala-containing precursors was required for teicoplanin resistance. The VanY and VanZ proteins also encoded by Tn1546 were not effectors of the regulation of the vanHAX operon but contributed to vancomycin and teicoplanin resistance, respectively. Differences at the regulatory level accounted for phenotypic diversity in acquired glycopeptide resistance by production of D-lac-ending precursors.

Anti-Bacterial Agents↗
PubMed2000

Selective pressure by antibiotic use in livestock.

Selective pressure exerted by the use of antibiotics as growth promoters in food animals appears to have created large reservoirs of transferable antibiotic resistance in these ecosystems. This first became evident for oxytetracycline and later for the streptothricin antibiotic nurseothricin, for which a transfer of relevant resistance determinants (sat genes) to bacterial pathogens of humans was demonstrated. With the emergence of glycopeptide resistance in Enterococcus faecium outside hospitals, a large reservoir of transferable resistance (vanA gene cluster) was identified in animal husbandry due to the use of avoparcin as feed additive. The spread of resistance, which reaches the human enterococcal flora via meat products, is probably due to the dissemination of the vanA gene cluster integrated into different conjugative plasmids among a variety of different strains. Streptogramin resistance associated with the resistance genes vatA and vatG has been found in E. faecium of animal and of clinical origin. Because virginiamycin has been used as growth promoter in animals but streptogramins have been used infrequently in human medicine, this again suggests an animal origin of resistance. Since the use of avoparcin ended, a decline in the rates of glycopeptide-resistant E. faecium (GREF) from animals and humans in the community has been recorded. This supports the ban of antibacterial growth promoters that might interfere with human chemotherapy that has been introduced in European Union countries.

Animals↗
PubMed2002

In vitro activity of AZD2563, a novel oxazolidinone, against European Gram-positive cocci.

AZD2563 is a new oxazolidinone that has targeted activity against Gram-positive bacteria. The in vitro activity of AZD2563 and nine comparators against 1543 European enterococcal, staphylococcal and streptococcal isolates was determined. The compound is a potent oxazolidinone, with no isolate tested displaying an MIC > 4 mg/L and 94.4% having an MIC < or =2 mg/L. Compared with linezolid, the distribution of MICs for all species of bacteria tested, with the exception of Enterococcus faecium, was shifted by one or two dilution steps to lower values for AZD2563. Thus, this oxazolidinone is a promising new antibiotic for the treatment of infections with Gram-positive cocci.

Anti-Bacterial Agents↗
PubMedDate not supplied

[The use of the polymerase chain reaction method for the identification of representatives of the genus Enterococcus].

The possibility of the use of repetitive extragenic palindrome (REP)-specific primers to PCR-fingerprint analysis of Enterococcus genomes has been shown for the first time. Presence of the similar PCR products, and first of all the fragment 950 bp in size, has been observed in all the strains studied. Differences in PCR-product composition displayed genetical heterogeneity between Enterococcus faecium strains. These data evidence for the wide distribution of the REP, and rather high level of homology of the REP sequences among enterobacterial genomes.

Animals↗
PubMed2006

Genes encoding bacteriocins and their expression and potential virulence factors of Enterococci isolated from wood pigeons (Columba palumbus).

Samples of the intestinal content and carcasses of wood pigeons (Columba palumbus) were evaluated for enterococci with antimicrobial activity. Enterococcus faecium comprised the largest enterococcal species with antagonistic activity, followed by Enterococcusfaecalis and Enterococcus columbae. PCR amplification of genes coding bacteriocins and determination of their nucleotide sequence, and the use of specific antipeptide bacteriocin antibodies and a noncompetitive indirect enzyme-linked immunosorbent assay, permitted characterization of enterococci coding that described bacteriocins and their expression. The efaAfm determinant was the only virulence gene detected in E. faecium, whereas E. faecalis showed a larger number of virulence determinants, and E. columbae did not carry any of the virulence genes examined. Although all E. faecalis isolates manifested a potent direct antimicrobial activity, no activity was detected in supernatants of producer cells. Purification of the antagonistic activity of E. columbae PLCH2 showed multiple chromatographic fragments after matrix-assisted laser desorption-ionization time-of-flight mass spectrometry analysis, suggesting the active peptide(s) had not yet purified to homogeneity. Bacteriocinogenic E. faecium and E. columbae isolates may be considered hygienic for production of enterocins and potentially safe due to their low incidence of potential virulence genes and susceptibility of most relevant clinical antibiotics. However, the presence among the enterococci of E. faecalis strains with a potent antagonistic activity and multiple virulence factors is an issue that must be considered further.

Animals↗
PubMed2003

Identification of potent and broad-spectrum antibiotics from SAR studies of a synthetic vancomycin analogue.

Dimeric vancomycin analogues based on a lead compound identified from a library of synthetic analogues of vancomycin have up to 60-fold greater activity than vancomycin against vancomycin-resistant Enterococcus faecium (VRE, VanA phenotype). Simplified analogues have also been prepared and found to maintain activity against VRE and have broad-spectrum antibiotic activity.

Anti-Bacterial Agents↗
PubMed2001

Pestalone, a new antibiotic produced by a marine fungus in response to bacterial challenge.

The isolation and structure determination of a new chlorinated benzophenone antibiotic, pestalone (1), is described. The new compound was produced by a cultured marine fungus only when a unicellular marine bacterium, strain CNJ-328, was co-cultured in the fungal fermentation. The fungus, isolated from the surface of the brown alga Rosenvingea sp. collected in the Bahamas Islands, was identified as an undescribed member of the genus Pestalotia. The structure of 1, initially assigned with only modest confidence by combined spectral and chemical data, was confirmed by single-crystal X-ray analysis. Pestalone (1) exhibits moderate in vitro cytotoxicity in the National Cancer Institute's 60 human tumor cell line screen (mean GI(50) = 6.0 microM). More importantly, pestalone shows potent antibiotic activity against methicillin-resistant Staphylococcus aureus (MIC = 37 ng/mL) and vancomycin-resistant Enterococcus faecium (MIC = 78 ng/mL), indicating that pestalone should be evaluated in advanced models of infectious disease.

Anti-Bacterial Agents↗
PubMed2001

Substrate specyfity of peptidases synthesised by Enterococcus facieum.

The strain Enterococcus faecium successfully used in youghurt production was the subject of studies. It results from the comparison of the activities of peptidases synthesised by E. faecium that aminopeptidases dominated Their average activity was about 2.5-fold higher than the average activity of dipeptidases. The presence of carboxy- and tripeptidases is also important from the technological point of view.

Amino Acid Sequence↗
PubMed1996

Vancomycin-resistant enterococcus: case reviews after transplantation.

Increased and inappropriate use of vancomycin during the past 35 years has led to the development of vancomycin-resistant enterococcal (VRE) species that are difficult and expensive to treat. This article presents case reviews of patients who experienced vancomycin-resistant enterococcus faecium (VREF) infections during prolonged hospitalizations after transplantation. The risk factors for the development of VRE infection, prevention measures, and the interventions to be implemented once VRE infection is diagnosed are discussed.

Adult↗
PubMed2004

[A case of probable bacterial translocation-associated sepsis in healthy adult].

We report a patient with bacterial translocation-associated sepsis who was healthy and did not have any related-background. The 57-year-old male had been well until 16 hours before admission, when nausea and vomiting gradually developed and increased in intensity. In the morning of May 22, 2002, he had shaking chills, temperature of 38.6 degrees C and watery diarrhea, and was admitted to Kawasaki Municipal Hospital. On admission, temperature was 40.7 degrees C but otherwise physical examination revealed no particular abnormality. Laboratory data showed total white blood cells of 28,400/microliter, platelet count of 130,000/microliter, creatinine of 2.0 mg/dl and C-reactive protein of 7.5 mg/dl. 1 g of cefmetazole was administered every eight hours. In the early morning of May 23, he suddenly went into shock. At that time, laboratory findings revealed total white blood cells of 33,700/microliter, platelet count of 65,000/microliter, C-reactive protein of 24.9 mg/dl, creatinine of 5.6 mg/dl and serum potassium concentration of 5.7 mEq/l. Gram positive cocci and gram negative rods were isolated from blood culture obtained on admission. Cefmetazole was changed to 1.5 g/day of imipenem/cilastatin sodium and 600 mg/day of clindamycin. In addition, hemodialysis and endotoxin removal with an adsorbent column using polymyxin B were performed. Bacteria detected in the blood on admission were identified as Klebsiela oxytoca and Enterococcus faecium. Imipenem/cilastatin sodium and clindamycin were continued for 13 days. The patient recovered fully and was discharged on June 11. This case suggests that bacterial translocation-associated sepsis might occur even in a hitherto healthy adult.

Bacterial Translocation↗