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Antimicrobial activity of everninomicin against clinical isolates of Enterococcus spp., Staphylococcus spp., and Streptococcus spp. tested by Etest.

The in-vitro antimicrobial activity of everninomicin, a novel oligosaccharide antibiotic, was tested against clinical isolates of 11 methicillin-susceptible Staphylococcus aureus (MSSA) strains, 11 methicillin-resistant S. aureus (MRSA) strains, 22 Staphylococcus epidermidis strains, 23 Enterococcus faecalis strains, 23 Enterococcus faecium strains, 23 Enterococcus avium strains, 27 Streptococcus pneumoniae strains, 22 Streptococcus pyogenes strains, and 20 Streptococcus agalactiae strains. The minimum inhibitory concentrations (MICs) of everninomicin were determined by Etest and compared with those of vancomycin, teicoplanin, and minocycline. The Etest showed that everninomicin exhibited excellent activity (MIC, <0.016 to 1.5 microg/ml) against the gram-positive cocci tested. The antimicrobial activity of everninomicin against S. aureus and S. epidermidis was stronger than that of vancomycin, teicoplanin, and minocycline, in particular against MRSA, with an MIC50 of 0.38 microg/ml and an MIC90 of 0.5 microg/ml. Against strains of enterococci and streptococci, everninomicin was more active than vancomycin, but as active as teicoplanin.

Aminoglycosides↗

Delayed-onset streptococcus pyogenes endophthalmitis following Ahmed glaucoma valve implantation.

BACKGROUND: To report a case of delayed-onset Streptococcus pyogenes endophthalmitis following implantation of an Ahmed glaucoma valve. CASE: A 10-year-old patient presented with acute endophthalmitis 1 year after Ahmed glaucoma valve implantation. OBSERVATIONS: The conjunctiva and Tenon's capsule over the valve plate had been penetrated by one of the polypropylene fixation sutures. The valve was removed, and pars plana vitrectomy was performed. Vitreous specimens and removal of the discharge over the plate revealed Streptococcus pyogenes. CONCLUSIONS: This is the first documented case of Streptococcus pyogenes endophthalmitis following Ahmed glaucoma valve implantation. We think the conjunctival buttonhole caused by the polypropylene suture provided an entry site for the infection.

Child↗

Perinatal mortality in Victoria, Australia: role of group B Streptococcus.

Group B beta-hemolytic streptococcus is the most common infective cause of neonatal morbidity and mortality. It is therefore surprising that no agreement exists for an approach to its prevention. There is also increasing evidence that occult infection may play an etiologic role in premature rupture of the membranes and preterm labor. In this report we review the role of group B beta-hemolytic streptococcal sepsis as a cause of perinatal wastage in the state of Victoria, Australia during the period 1982 to 1987. Group B beta-hemolytic streptococcus accounted for 1.7% of overall perinatal deaths, and for 30.3% (77 of 254) perinatal deaths directly attributable to infection. By comparison, over the same 6-year period, erythroblastosis accounted for 0.5% of perinatal wastage and there were only two deaths as a result of congenital syphilis. The true incidence of lethal group B beta-hemolytic streptococcal infection is probably greater because of the absence of histologic and bacteriologic studies in many perinatal deaths. We believe that intrapartum chemoprophylaxis with penicillin of all group B beta-hemolytic streptococcus-positive carrier mothers would significantly reduce neonatal morbidity and mortality from this cause.

Cause of Death↗

Microbial surface interactions: reduction of the haemagglutination activity of the oral bacterium Fusobacterium nucleatum by absorption with Streptococcus and Bacteroides.

Oral strains of Fusobacterium nucleatum showed haemagglutination (HA) of sheep red blood cells and attachment of HA-active F. nucleatum fragments to other microorganisms allowed a means of studying microbial surface interactions. HA-active sonicated fragments (SF) prepared from F. nucleatum were mixed with whole cell suspensions of 48 bacterial strains and, after incubation, the whole cells were separated from the non-absorbed fragments by differential centrifugation. Attachment of F. nucleatum fragments to the cells was indicated by a reduction in the HA activity of the SF in the supernatant fluid remaining after absorption with whole cells. HA activity of the microbial cells used for absorption and the detection of F. nucleatum fragments on these cells by an indirect fluorescent antibody technique provided further evidence of attachment. Of the 48 strains tested, 10 absorbed F. nucleatum HA-active fragments. They included Bacteroides gingivalis, Bacteroides fragilis subsp. distasonis, Bacteroides corrodens, Streptococcus morbillorum, Streptococcus sanguis (Blackburn and JC 74) and Streptococcus mutans AHT, BHT, 10449 and 6715. Chelators revealed that F. nucleatum attached to the microorganisms via a Ca2+-dependent interaction. Sugar inhibition demonstrated that F. nucleatum attached to the microorganisms via a D-galactose-containing moiety on their surface. A reduction in the absorption of F. nucleatum HA-active fragments by Strep. mutans grown in a higher concentration of sucrose was observed.

Absorption↗

The effect of smokeless-tobacco extracts on the growth of oral bacteria of the genus Streptococcus.

Aqueous tobacco extracts were used to supplement a basic salts solution (BSS) and a microbial medium. Thin-layer chromatography revealed sucrose in only one of four extracts. Discs saturated with extracts (0.1-50 mg/ml) failed to inhibit growth of any of the micro-organisms. Supplementation (10 mg/ml) of BSS with the tobaccos lacking sucrose resulted in augmented growth of Streptococcus mutans, Streptococcus salivarius and Streptococcus sanguis whereas the sucrose-containing brand augmented only Strep. sanguis growth. Thus extracts of these smokeless tobaccos would serve as a growth substrate for three species of oral streptococci which are frequently associated with human dental caries.

Bacteriological Techniques↗

Binding studies of macrolides, lincosamides and streptogramins to Streptococcus G group using [3H]-erythromycin.

Parameters of [3H]-erythromycin binding to Streptococcus are determined in vivo using both equilibrium and kinetic methods. This binding is saturable, reversible and independent of energetic systems. Whatever the methods used, the binding parameters are identical as 14 nM for the dissociation constant of the complex erythromycin-Streptococcus and a density of binding sites of 11,865 molecules/cell. Other macrolides, streptogramins and lincosamides competitively displaced bound [3H]-erythromycin suggesting that these compounds share common binding sites on the bacteria. In parallel, the MIC values of these antibiotics against Streptococcus are determined by agar dilution method in Mueller-Hinton medium with 5% of horse blood in order to compare the binding and microbiological parameters. A strong correlation (n = 0.863) has been found between the corresponding inhibition constants and MIC values. Such binding studies could be used in conjunction with microbiological assays for primary screening of active analogous or other compounds with interfere with [3H]-erythromycin binding to the bacteria.

Anti-Bacterial Agents↗

Purification and characterization of recombinant cell surface protein antigen A of Streptococcus sobrinus B13N.

1. It has been reported that immunization of rhesus monkeys with the surface protein antigen I/II from Streptococcus mutans significantly reduced dental caries. 2. The surface protein antigen A (SpaA) from Streptococcus sobrinus is known to correspond antigenically to I/II. MD51 is an Escherichia coli host containing pMD51, a plasmid encoding the SpaA gene from Streptococcus sobrinus B13N. 3. The recombinant SpaA (rSpaA) was purified from cell extracts of Escherichia coli clone MD51. 4. The purified recombinant SpaA was homogeneous with a molecular weight of 210 kDa according to SDS-PAGE and had an isoelectric point of 4.2 based on isoelectric focusing. 5. Amino acid composition of rSpaA showed a relatively high amount of hydrophobic amino acids (39.7%).

Amino Acids↗

Enhanced antenatal detection of group B streptococcus colonization.

OBJECTIVE: To improve culture methods for the detection of group B streptococcus colonization. METHODS: This study prospectively compared the standard culture medium, a blood agar plate, to a selective culture medium, Todd Hewitt broth with antibiotics, and compared vaginal culture with rectal culture at the first prenatal exam. RESULTS: Of the 383 vaginal swabs received for evaluation of the two culture media, 78 (20.4%) were positive for group B streptococcus. The detection rates of the blood agar plate method and the Todd Hewitt broth with antibiotics were 64.1 and 97.4%, respectively. Using the Todd Hewitt broth with antibiotics, an additional 94 patients were cultured vaginally and rectally. Twenty-nine (30.9%) had positive cultures. The rate of detection was 58.6% for the vaginal culture, 89.7% for the rectal culture, and 100% for both culture sites combined. CONCLUSION: These data indicate that culture detection of group B streptococcus can be improved by using both a selective broth medium and a dual vaginal and rectal culture.

Carrier State↗

Evaluation of the modified Visuwell Strep-A enzyme immunoassay for detection of group-A Streptococcus from throat swabs.

The modified Visuwell Strep-A enzyme immunoassay (EIA) was compared with culture for detection of group-A Streptococcus from throat swabs. Throat swabs in modified Stuarts medium obtained after culture at two institutions were tested in Visuwell. Cumulative results were n = 417, sensitivity 87.8%, specificity 89.9% predictive value positive (PVP) 67.9%, predictive value negative (PVN) 96.8%, and accuracy 89.5%. At another site, swabs were delivered to the laboratory without transport medium, cultured, and subsequently tested by Visuwell (n = 202, sensitivity 79.6%, specificity 84.5%, PVP 65.2%, PVN 91.9%, accuracy 83.2%). When 1+ culture-positive specimens were considered negative, the sensitivity and PVN increased from 79.6% to 90.2% and 91.9% to 97.1% respectively. Overall performance of the modified Visuwell was comparable with that of the initial assay for throat swabs transported with or without modified Stuarts medium. Cross reaction with organisms other than group-A Streptococcus normally found in the oropharynx was negligible in Visuwell and the limit of detection of group-A Streptococcus was 5 x 10(4) colony-forming units.

Cross Reactions↗

Molecular characterization and functional analysis of the manganese-containing superoxide dismutase gene (sodA) from Streptococcus thermophilus AO54.

This report describes the isolation, sequencing, and functional analysis of the sodA gene, encoding Mn-superoxide dismutase, from Streptococcus thermophilus AO54. The gene was found to encode a 201 amino acid polypeptide with 88 and 83% identity to SodA from Streptococcus mutans and Streptococcus agalacticae, respectively. Primer extension analysis revealed a transcriptional start site 27 nucleotides upstream of initiation codon. The gene was expressed in Escherichia coli and was able to rescue the growth of a sodAsodB mutant in a minimal-medium containing 10(-6)M paraquat. A sodA mutant of S. thermophilus was constructed and found to be more sensitive to aerobic growth than its parent strain. Supplementing the medium with MnCl(2) improved the growth of the mutant, only under microaerophilic conditions. The results suggest that sodA is essential for the aerobic growth of S. thermophilus. In the absence of functional SodA, manganese ions may provide partial protection against oxygen toxicity.

Amino Acid Sequence↗

Evaluation of the BD Phoenix Automated Microbiology System SMIC/ID panel for identification and antimicrobial susceptibility testing of Streptococcus spp.

The BD Phoenix Automated Microbiology System SMIC/ID panel was evaluated for identification and antimicrobial susceptibility testing (AST) of various streptococci. A group of 97 consecutive clinical isolates of Streptococcus pneumoniae, 23 Streptococcus pyogenes, 24 Streptococcus agalactiae, and 34 viridans streptococci were collected and comparisons made with routine manual methods used in the clinical microbiology laboratory. Overall, in 162 (91%) of 178 isolates, Phoenix identification results demonstrated agreement. For AST results for the 162 isolates that demonstrated identification concordance, the overall essential agreement rate was 98.5%; the category agreement was 94.9%; and the very major error, major error, and minor error rates were 0%, 0.15%, and 5.8%, respectively. Although relatively high minor error rates were observed with S. pneumoniae and beta-lactams, 79.2% of the 77 minor errors were the result of a single log(2) dilution difference. The Phoenix SMIC/ID panel performed favorably and demonstrated the advantages of automation and simple methodology.

Anti-Bacterial Agents↗

Lack of mitogenic activity of speG- and speG(dys)-positive Streptococcus dysgalactiae subspecies equisimilis isolates from patients with invasive infections.

In recent years, Streptococcus dysgalactiae subspecies equisimilis has been isolated with an increasing frequency as the cause of invasive streptococcal diseases. For 46 S. dysgalactiae subspecies equisimilis isolates from invasive infections and four isolates from superficial infections, the presence of emm/emmL genes and of genes encoding various different streptococcal superantigens was determined by polymerase chain reaction (PCR). Subsequently, PCR products were identified by DNA sequencing, and the expression of mRNA from superantigen genes was assessed by reverse transcriptase-PCR. The mitogenic activity of S. dysgalactiae subspecies equisimilis was assessed by [3H]thymidine incorporation into human lymphocytes and compared with that of Streptococcus agalactiae and Streptococcus pyogenes. All S. dysgalactiae subspecies equisimilis isolates studied harbored an emm/emmL gene. Only in six of the S. dysgalactiae subspecies equisimilis isolates from invasive infections, speG was detected by PCR, two of which were further identified as speGdys by sequencing of the PCR product. None of the S. dysgalactiae subspecies equisimilis isolates harbored any of the genes speA, speB, speC, speF, speH, speI, speJ, speK, speL, speM, smeZ, or ssa of S. pyogenes. In contrast to S. pyogenes, no expression of speG or speGdys mRNA, respectively, was detected in the reverse transcriptase-PCR assay for any of the speG- or speGdys-positive S. dysgalactiae subspecies equisimilis isolates. Moreover, S. dysgalactiae subspecies equisimilis and S. agalactiae revealed no or very low mitogenic activity, while S. pyogenes was a very powerful inducer of proliferative responses. These findings support the hypothesis that the pathogenicity of S. dysgalactiae subspecies equisimilis may be associated in part with the presence of emm/emmL genes, and suggest that the severity of S. dysgalactiae subspecies equisimilis invasive infections is not mediated by superantigen-induced mitogenicity.

Antigens, Bacterial↗

A GapC chimera retains the properties of the Streptococcus uberis wild-type GapC protein.

The GapC products of Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis share considerable homology at the DNA and amino acid levels. The high similarity at the protein level suggests that one GapC protein might be used as a single antigen to protect dairy cows against infections with the contagious S. agalactiae and the environmental S. dysgalactiae and S. uberis strains. Despite their similarities, immunization with the S. dysgalactiae GapC did not protect dairy cows from a challenge with S. uberis, suggesting the presence of regions in GapC that are involved in species-specific protection. To produce a single antigen that can be used to protect against all streptococcal mastitis infections, we constructed a GapC chimeric protein using the S. uberis GapC product as the backbone followed by non-conserved peptide regions from the S. agalactiae and S. dysgalactiae GapC proteins. We report that the chimeric GapC protein retains the enzymatic activity of the S. uberis GapC protein. In addition, we fused the chimera to the OmpF and LipoF transport sequences of Escherichia coli and the GapC chimeras were present in membrane fractions of E. coli. These extracts could be the basis of an antigen preparation for use in mastitis vaccines.

Amino Acid Sequence↗

Efficacy of Streptococcus agalactiae (group B) vaccine in tilapia (Oreochromis niloticus) by intraperitoneal and bath immersion administration.

We evaluated the effectiveness of a Streptococcus agalactiae vaccine in tilapia (Oreochromis niloticus) for prevention of streptococcal disease. The vaccine was prepared from formalin-killed cells and concentrated extracellular products (greater than 3 kDa) of a single isolate of S. agalactiae (ARS-KU-MU-11B). Intraperitoneal (IP) and bath immersion (BI) vaccine trials were conducted at two temperatures, 32 and 26 degrees C, and mean fish weights, 5 and 30 g. Control tilapia were injected with tryptic soy broth. Thirty gram tilapia vaccinated and challenged by IP injection with 1.5 x 10(4) colony-forming units (CFU)/fish of Streptococcus agalactiae at 30 days post-immunization had a relative percent survival (RPS) of 80. Smaller tilapia vaccinated and challenged under similar conditions had an RPS of 25. An RPS of zero was noted in 30 g fish IP vaccinated with Streptococcus iniae and IP challenged with S. agalactiae. The 5 and 30 g tilapia bath immunized with S. agalactiae and IP challenged with 3.6 x 10(5) and 1.7 x 10(6) CFU/fish of S. agalactiae had RPS values of 34. Intraperitoneal administration of the vaccine provided efficacious protection only in the 30 g tilapia regardless of whether the fish were immunized and challenged at 26 or 32 degrees C. Bath immunization of both 5 and 30 g tilapia resulted in RPS values that were two times lower than those achieved with IP vaccination. The results of this study suggest that there is a lack of cross-protection of S. iniae bacterins against S. agalactiae challenge. Protection against S. agalactiae infection is, however, provided through vaccination with a S. agalactiae modified bacterin vaccine.

Animals↗

Identification of Streptococcus spp. causing bovine mastitis by PCR-RFLP of 16S-23S ribosomal DNA.

Traditional microbiological methods for identification of Streptococcus spp. causing bovine mastitis have been demonstrated to be less than highly reliable. PCR-RFLP analysis of 16S-23S ribosomal DNA was used to characterise seven reference strains of streptococcal mastitis pathogens as well as four reference strains of other gram-positive, catalase-negative cocci of bovine origin to allow comparative identification of field isolates. RFLP analysis of PCR products, using a combination of two restriction endonucleases in single reactions (HaeIII and AluI, HaeIII and RsaI or AluI and RsaI) generated unique patterns for species of Streptococcus, Enterococcus and Lactococcus. One hundred field isolates of Streptococcus spp. collected from cows with clinical or subclinical mastitis were tested. Fifty-seven isolates, classified by conventional tests as S. uberis, were identified as 47 S. uberis and six S. parauberis by their unique RFLP patterns. The remaining four isolates had RFLP patterns distinct from the reference strains and two of these were identified as closely related to S. iniae and two to Aerococcus viridans by 16S rRNA sequencing. Conventional identification of 17 S. agalactiae and 26 S. dysgalactiae subsp. dysgalactiae was confirmed by RFLP. Use of a combination of restriction enzymes in a single tube enabled the rapid, accurate, cost effective and easily performed identification of all major streptococcal mastitis pathogens.

Animals↗

[Necrotizing subcutaneous infection by Streptococcus agalactiae].

Necrotizing soft tissue infections constitute some of the most potentially threatening infections that may be acquired in the community or in the hospital milieu as they are associated with a high mortality rate. In most cases they are produced by Streptococcus pyogenes. We report a case of a necrotizing soft tissue infection caused by Streptococcus agalactiae (group B beta hemolytic streptococcus) that involved the leg of an elderly man with chronic lymphatic leukemia and diabetes mellitus. The lesions notably improved after initiating intravenous antibiotic treatment with amoxicillin-clavunate and clindamycin.

Aged↗

Effects of alterations of zinc-to-phosphorus ratios and meconium content on group B Streptococcus growth in human amniotic fluid in vitro.

Several authors have described the bacterial growth-altering properties of amniotic fluid. We examined Group B Streptococcus growth in aseptically obtained amniotic fluid in vitro after altering its zinc, phosphorus, and meconium contents. Zinc and phosphorus levels were calculated in amniotic fluid and in meconium. Separate solutions of zinc and phosphorus were added to yield concentrations of 0.7, 7.0, 70, and 700 mumol. The solutions were incubated with Group B Streptococcus III 893 and Escherichia coli C5 strains, and 24-hour growth curves were plotted. Meconium, 0.5 mg/ml, was added to each amniotic fluid + zinc and amniotic fluid + phosphorus solution, and growth curves were plotted. The rate of proliferation of Group B Streptococcus varied directly with the zinc concentration (700 = 70 greater than 7 greater than 0.7 mumol) and inversely with the phosphorus content (700 less than 70 less than 7 = 0.7 mumol). Meconium enhanced the proliferative effect of zinc and hindered the inhibitory effect of phosphorus. Thus one possible mechanism whereby meconium enhances bacterial growth in amniotic fluid may be by alteration of zinc-to-phosphorus ratios.

Amniotic Fluid↗

Brain abscess caused by Streptococcus intermedius: two case reports.

BACKGROUND: Although Streptococcus intermedius has been recognized as an important pathogen for abscess formation outside the central nervous System, the Streptococcus milleri group, of which it is a member, has not been recognized as a specific pathogen for brain abscess, often thought to be caused by Streptococcus viridans, which includes other streptococcal species. CASE DESCRIPTION: Two cases of brain abscess in the left frontal lobe caused by S. intermedius, which responded well to antimicrobial treatment combined with needle aspiration, are presented. In the first patient, the predisposing disease was paranasal sinusitis of the frontal and ethmoid sinuses. In the second patient, the source of the pathogen was not detected despite extensive examination. The patients underwent aspiration of pus under ultrasound guidance in the first patient, and via a computed tomography-guided stereotactic procedure in the second patient. They subsequently received appropriate antimicrobial therapy against S. intermedius isolated from the pus culture. Both patients were discharged without any neurological deficits. CONCLUSION: It is important to recognize S. intermedius as a pathogen of brain abscess, and to be aware of its predisposing factors, i.e., mucosal disturbance and liver abscess.

Adolescent↗