Experimental study on the quasi-steady approximation of glottal flows.

To examine the quasi-steady approximation of the glottal flow, widely used in the modeling of vocal fold oscillations, intraglottal pressure distributions were measured in a scaled-up static vocal fold model under time-varying flow conditions. The left and right vocal folds were slightly open and set to a symmetric and oblique configuration with a divergence angle. To realize time-varying flow conditions, the flow rate was sinusoidally modulated with a frequency of 2 and 10 Hz, which correspond to 112.5 and 562.5 Hz, respectively, in real life. Measurements of the intraglottal pressures under both steady and time-varying flows revealed that the pressure profiles of the time-varying flow conditions are non-distinguishable from those of the steady flow conditions as far as they have the same subglottal pressure as an input pressure. The air-jet separation point was also non-distinguishable between the steady and the time-varying flow conditions. Our study therefore suggests that the time-varying glottal flow can be approximated as a series of steady flow states with a matching subglottal pressure in the range of normal vocalization frequencies. Since the glottal closure was not taken into account in the present experiment, our argument is valid except for such a critical situation.

Influence of level difference due to vocal folds angular asymmetry on auto-oscillating replicas.

Dysphonia is often caused by level difference between left and right vocal folds, which are positioned on different angles with respect to the transverse plane, resulting in angular asymmetry. Unilateral vocal fold paralysis may cause such angular asymmetry. In this case, the normal vocal fold is located on the transverse plane, whereas the paralyzed vocal fold is rotated in the sagittal plane as its posterior edge is moved up to the superior direction. The effect of such angular asymmetry (up to 25°) between the left and right vocal fold on the auto-oscillation is experimentally studied using mechanical replicas. For all replicas, it is observed that, as full contact between vocal folds is lost, increase of angular asymmetry results in a decrease of the signal-to-noise ratio, an increase of the total harmonic distortion rate, and an increase of the oscillation threshold pressure. These general tendencies are in agreement with clinical findings reported for vertical level difference during phonation. In analogy to the preceding experimental study in which vocal folds are spaced in parallel with a vertical trade-off, a formula is proposed to describe the oscillation threshold as a function of angular asymmetry.

Adult invasive and noninvasive infections due to Streptococcus dysgalactiae subsp. equisimilis in France from 2006 to 2010.

We characterized 182 Streptococcus dysgalactiae subsp. equisimilis isolates and analyzed the epidemiological data on the corresponding infections. stG6, stG485, and stG6792 were the 3 most prevalent invasive emm types among the 27 different emm types recovered. High rates of antimicrobial resistance were observed for macrolides (26.4%) and tetracycline (34.6%).

Preeminence of Staphylococcus aureus in infective endocarditis: a 1-year population-based survey.

BACKGROUND: Observational studies showed that the profile of infective endocarditis (IE) significantly changed over the past decades. However, most studies involved referral centers. We conducted a population-based study to control for this referral bias. The objective was to update the description of characteristics of IE in France and to compare the profile of community-acquired versus healthcare-associated IE. METHODS: A prospective population-based observational study conducted in all medical facilities from 7 French regions (32% of French individuals aged ≥18 years) identified 497 adults with Duke-Li-definite IE who were first admitted to the hospital in 2008. Main measures included age-standardized and sex-standardized incidence of IE and multivariate Cox regression analysis for risk factors of in-hospital death. RESULTS: The age-standardized and sex-standardized annual incidence of IE was 33.8 (95% confidence interval [CI], 30.8-36.9) cases per million inhabitants. The incidence was highest in men aged 75-79 years. A majority of patients had no previously known heart disease. Staphylococci were the most common causal agents, accounting for 36.2% of cases (Staphylococcus aureus, 26.6%; coagulase-negative staphylococci, 9.7%). Healthcare-associated IE represented 26.7% of all cases and exhibited a clinical pattern significantly different from that of community-acquired IE. S. aureus as the causal agent of IE was the most important factor associated with in-hospital death in community-acquired IE (hazard ratio [HR], 2.82 [95% CI, 1.72-4.61]) and the single factor in healthcare-associated IE (HR, 2.54 [95% CI, 1.33-4.85]). CONCLUSIONS: S. aureus became both the leading cause and the most important prognostic factor of IE, and healthcare-associated IE appeared as a major subgroup of the disease.

Globicatella sanguinis meningitis associated with human carriage.

Globicatella sanguinis is a rare cause of acute meningitis. We demonstrated human carriage of Globicatella by identifying cefotaxime-resistant strains in groin and rectal specimens 9 months after invasive infection. The pathogenic strain isolated from the cerebrospinal fluid and the carriage strains were accurately identified by sodA gene sequence analysis.

Clinical and microbiological characteristics of severe Streptococcus pyogenes disease in Europe.

In an attempt to compare the epidemiology of severe Streptococcus pyogenes infection within Europe, prospective data were collected through the Strep-EURO program. Surveillance for severe cases of S. pyogenes infection diagnosed during 2003 and 2004 was undertaken in 11 countries across Europe by using a standardized case definition and questionnaire. Patient data as well as bacterial isolates were collected and characterized by T and M/emm typing, and selected strains were analyzed for the presence of superantigen genes. Data were analyzed to compare the clinical and microbiological patterns of the infections across the participating countries. A total of 4,353 isolates were collected from 5,521 cases with severe S. pyogenes infections who were identified. A wide diversity of M/emm types (n = 104) was found among the S. pyogenes clinical isolates, but the M/emm type distribution varied broadly between participating countries. The 10 most predominant M/emm types were M/emm type 1 (M/emm1), M/emm28, M/emm3, M/emm89, M/emm87, M/emm12, M/emm4, M/emm83, M/emm81, and M/emm5, in descending order. A correlation was found between some specific disease manifestations, the age of the patients, and the emm types. Although streptococcal toxic shock syndrome and necrotizing fasciitis were caused by a large number of types, they were particularly associated with M/emm1 and M/emm3. The emm types included in the 26-valent vaccine under development were generally well represented in the present material; 16 of the vaccine types accounted for 69% of isolates. The Strep-EURO collaborative program has contributed to enhancement of the knowledge of the spread of invasive disease caused by S. pyogenes within Europe and encourages future surveillance by the notification of cases and the characterization of strains, which are important for vaccination strategies and other health care issues.

Acute appendicitis, an unusual presentation of Streptococcus pyogenes infection.

The incidence of invasive streptococcal pyogenic infections has been on a worldwide increase. The most frequent clinical manifestations start with cutaneous septicemias. We report an unusual case of Streptococcus pyogenes-based septicemia, secondary to appendicitis with rapidly evolving peritonitis.

Epidemiology of severe Streptococcus pyogenes disease in Europe.

The past 2 decades have brought worrying increases in severe Streptococcus pyogenes diseases globally. To investigate and compare the epidemiological patterns of these diseases within Europe, data were collected through a European Union FP-5-funded program (Strep-EURO). Prospective population-based surveillance of severe S. pyogenes infection diagnosed during 2003 and 2004 was undertaken in 11 countries across Europe (Cyprus, the Czech Republic, Denmark, Finland, France, Germany, Greece, Italy, Romania, Sweden, and the United Kingdom) using a standardized case definition. A total of 5,522 cases were identified across the 11 countries during this period. Rates of reported infection varied, reaching 3/100,000 population in the northern European countries. Seasonal patterns of infection showed remarkable congruence between countries. The risk of infection was highest among the elderly, and rates were higher in males than in females in most countries. Skin lesions/wounds were the most common predisposing factor, reported in 25% of cases; 21% had no predisposing factors reported. Skin and soft tissue were the most common foci of infection, with 32% of patients having cellulitis and 8% necrotizing fasciitis. The overall 7-day case fatality rate was 19%; it was 44% among patients who developed streptococcal toxic shock syndrome. The findings from Strep-EURO confirm a high incidence of severe S. pyogenes disease in Europe. Furthermore, these results have identified targets for public health intervention, as well as raising awareness of severe S. pyogenes disease across Europe.

Streptococcus sinensis: an emerging agent of infective endocarditis.

The identification by conventional methods of viridans streptococcal species, which are rarely encountered clinically, requires confirmation by genomic methods. We characterized a strain of Streptococcus sinensis responsible for infective endocarditis by sequencing both the 16S rRNA and the manganese-dependent superoxide dismutase genes.

Molecular characterization of Streptococcus pyogenes isolates to investigate an outbreak of puerperal sepsis.

OBJECTIVE: To describe microbiological characteristics and epidemiologic features of an outbreak of postpartum endometritis. METHODS: Various markers were investigated in five patients and three throat carriage isolates of Streptococcus pyogenes obtained during an outbreak of endometritis occurring in a 13-week period. Molecular characterization included biotyping, T-serotyping, emm gene sequence and restriction, pulsed-field gel electrophoresis (PFGE), and random amplified polymorphic DNA (RAPD) analysis. RESULTS: Biotype, T-serotype, and genotypic data (emm analysis, PFGE, and RAPD analysis) revealed a close relationship among the isolates from three patients, suggesting that cross-contamination had occurred. These isolates were biotype 1, T type 28, and emm type 28. The isolates from one patient and one carrier differed from those of the index patient by minor variations of the emm amplicon restriction pattern, PFGE pattern, or RAPD pattern. The remaining isolates were phenotypically and genetically different. CONCLUSION: Identification of different isolates demonstrated that different strains may circulate simultaneously during a true outbreak and that the predominant strain might persist for several months.

Impact of susceptibility to antibiotics of streptococci & enterococci isolated from patients with infective endocarditis on antibiotic treatment.

BACKGROUND & OBJECTIVES: Streptococci and enterococci are the most frequent pathogens causing infective endocarditis. In order to update the recommendations for both curative and prophylaxis treatment, the susceptibility to antibiotics of the most prevalent species of Streptococcaceae isolated from the patients with infective endocarditis was determined. METHODS: Streptococcal and enterococcal isolates (n=133) isolated from confirmed cases of infective endocarditis during a one-year prospective survey conducted in 1999 in France were studied. The identification of 106 streptococci and 27 enterococci to the species level was carried out by conventional methods. Their susceptibility to ten antibiotics used in curative or prophylactic treatment was measured. Minimal inhibitory concentrations were determined by agar dilution method. RESULTS: All the streptococcal and enterococcal isolates were susceptible to 4 mg/l or less of penicillin or amoxicillin. High levels of resistance to aminoglycosides were observed in two species, Streptococcus gallolyticus subsp. gallolyticus and Enterococcus faecalis. All isolates were susceptible to glycopeptides. Resistance to erythromycin, clindamycin, and pristinamycin was restricted to some species. INTERPRETATION & CONCLUSION: Curative treatments recommended for streptococcal or enterococcal endocarditis, including penicillin, amoxicillin or vancomycin in association with gentamicin were found to be appropriate for 98.5 per cent of cases. The emergence of erythromycin resistance in oral streptococci led to the use of pristinamycin in oral prophylactic treatment in patients allergic to beta-lactams.

Epidemiological markers of group A streptococcal infections in France.

BACKGROUND & OBJECTIVES: A limited number of biotypes, T-types, and emm-types have been found to be associated with invasive isolates of group A streptococci, confirming the involvement of the M protein in virulence and its importance as an epidemiological marker. In this study, the epidemiological markers in the clinical isolates of group A streptococci were compared in invasive and non invasive isolates. METHODS: From 1998 to 2001, 141 invasive and 353 non invasive isolates in France were studied and their biotype, T-type, and emm-type were determined. RESULTS: The invasive isolates were mostly obtained from blood whereas the non invasive isolates were isolated from throat. Most of the isolates were of biotype 1. The invasive isolates were mostly of the T-type 1 associated with emm-type 1. The T-type 4 associated with emm-type 4 and the T-type 28 associated with emm-type 28 were also frequent. Invasive isolates responsible for puerperal sepsis and non invasive isolates were mostly of the T-type 28 associated with emm-type 28. INTERPRETATION & CONCLUSION: This study confirms the high prevalence of isolates of biotype 1, T-type 1, and emm-type 1 among invasive isolates of group A streptococci. The emm-type 28 associated with T-type 28 was frequently observed in non-invasive isolates. A prospective study is being conducted to update the prevalence of the different emm-types in France, which will be of importance for the development of future vaccines.

New group D streptococcal species.

BACKGROUND & OBJECTIVES: The heterogeneity of group D streptococci led to the identification of various biotypes of Streptococcus equinus and Streptococcus bovis and to the description of new species. The objective of the present study was to improve the phenotypic delineation between species and to clarify their respective phylogenetic position. METHODS: Physiological and genomic analyses were carried out in 84 representative strains of the group D streptococci. Biotypes were determined with the API 20 strep and rapid ID 32 STREP systems of identification. Quantitative DNA-DNA hybridization under stringent conditions and values of the deltaT(m) allowed to delineate species and subspecies. The phylogenic position of the different genomic groups was determined by comparing the sequences of their 16S rDNA. RESULTS: Four DNA-clusters, including seven species or subspecies, were characterized. Differential associations of biochemical characters allowed their identification. S. equinus and the type strain of S. bovis belonged to a single species. S. gallolyticus, S. bovis biotype II.2, and S. macedonicus formed a single DNA-cluster including three different subspecies. These were designated as S. gallolyticus subsp. gallolyticus, S. gallolyticus subsp. pasteurianus, and S. gallolyticus subsp. macedonicus. The two other DNA-clusters corresponded to the two subspecies of S. infantarius, and to S. alactolyticus. INTERPRETATION & CONCLUSION: This study presented a new classification associated with an identification scheme of group D streptococci. The changes in this classification demonstrate the interest of a polyphagic approach of the bacterial identification.

Molecular mechanisms of tetracycline and macrolide resistance and emm characterization of Streptococcus pyogenes isolates in Tunisia.

Streptococcus pyogenes or group A Streptococcus, a major human pathogen, remains susceptible to beta-lactams, but resistance to other antibiotics is becoming more common. The purpose of this study was to characterize both phenotypic and genotypic epidemiological markers of group A Streptococcus and to identify the mechanisms of resistance to macrolides and tetracyclines. A total of 103 strains, isolated at Charles Nicolle University Hospital of Tunis, were investigated. The rate of resistance to erythromycin was low (5%), whereas a high rate of tetracycline resistance was found (70%). All the macrolide-resistant isolates expressed the constitutive macrolide, lincosamide, and streptograminB phenotype and harbored the erm(B) gene. Resistance to tetracycline was mainly due to the tet(M) gene, which is commonly associated with the conjugative transposon Tn916. No significant association was found between erm(B) and tet(M) genes. The tetracycline-resistant strains belonged to 28 distinct emm types. Among them, emm118 was the most prevalent type, followed by emm42, std432, emm76, and emm18. However, emm1, emm4, emm6, emm28, and emm3 were the most frequent types among tetracycline susceptible isolates. Only emm118 and emm42 types (p ≤ 0.05) were significantly associated with resistance to tetracycline.

Streptococcus henryi sp. nov. and Streptococcus caballi sp. nov., isolated from the hindgut of horses with oligofructose-induced laminitis.

Four Gram-positive, catalase-negative, coccoid-shaped isolates were obtained from the caecum and rectum of horses with oligofructose-induced equine laminitis. Phenotypic and phylogenetic studies were performed on these isolates. Initial biochemical profiling assigned two of the isolates to Streptococcus bovis. The other two isolates, however, could not be assigned conclusively on the basis of their biochemical profiles. Gene sequence analysis demonstrated that the four new isolates were related most closely to Streptococcus suis based on the 16S rRNA gene and to Streptococcus orisratti based on the manganese-dependent superoxide dismutase gene (sodA). Sequence divergence values from recognized Streptococcus species based on these two genes were >3 and >13%, respectively, for all four isolates. Phylogenetic and phenotypic analyses demonstrated that the four isolates formed two distinct clonal groups that are suggested to represent two novel species of the genus Streptococcus. The names proposed for these organisms are Streptococcus henryi sp. nov. (type strain 126(T) =ATCC BAA-1484(T) =DSM 19005(T)) and Streptococcus caballi sp. nov. (type strain 151(T) =ATCC BAA-1485(T) =DSM 19004(T)).

Prospective evaluation of a polymerase chain reaction-ELISA targeted to Aspergillus fumigatus and Aspergillus flavus for the early diagnosis of invasive aspergillosis in patients with hematological malignancies.

BACKGROUND: Current laboratory and radiological methods for diagnosis of invasive aspergillosis (IA) lack sensitivity and specificity. METHODS: We prospectively evaluated the diagnostic value of twice-weekly screening for circulating Aspergillus fumigatus and A. flavus DNA with a polymerase chain reaction-enzyme-linked immunosorbent assay (PCR-ELISA). RESULTS: Among the 201 adult patients with hematological malignancies who were included in the study, 55 IA cases were diagnosed. On the basis of the analysis of 1205 serum samples from 167 patients, the sensitivity, specificity, and positive and negative predictive values of the PCR-ELISA for proven and probable IA cases were 63.6%, 89.7%, 63.6%, and 89.7%, respectively, when samples with 2 consecutive positive results were used. The use of a combination of the PCR-ELISA and a galactomannan (GM) assay increased the sensitivity to 83.3%, increased the negative predictive value to 97.6%, and decreased the specificity to 69.8%. In most patients with IA, PCR-ELISA positivity anticipated or was simultaneous with the initiation of antifungal therapy, the abnormalities found by computed tomography, the mycological/histological diagnosis, and the GM positivity. Overall, 56.3% of the patients had at least 1 positive sample, and the false single-positive rate was 44.8%. CONCLUSIONS: In addition to serial screening for GM antigenemia and radiological surveillance, PCR-ELISA may improve the rates of early diagnosis of IA and the management of patients with hematological malignancies.

Bacteremia caused by an undescribed species of Janibacter.

A yellow-pigmented rod- to coccoid-shaped coryneform microorganism was isolated from the blood of a patient with acute myeloid leukemia. It was identified by 16S rRNA gene sequencing as a previously undescribed species of Janibacter. The isolate was susceptible to penicillins, aminoglycosides, fluoroquinolones, and glycopeptides.

Clonal spread of emm type 28 isolates of Streptococcus pyogenes that are multiresistant to antibiotics.

Fifty-three pharyngitis-related and invasive isolates of Streptococcus pyogenes that are resistant to bacitracin were collected. They were also resistant to streptomycin, kanamycin, macrolides, lincosamides, and streptogramin B. These multiresistant isolates were of emm type 28 and clonally related as shown by pulsed-field gel electrophoresis.

Identification of major Streptococcal species by rrn-amplified ribosomal DNA restriction analysis.

Amplified ribosomal DNA restriction analysis (rrn-ARDRA) is based on PCR amplification and restriction of a fragment of rRNA genes including 16S and 23S genes and the intergenic spacer. rrn-ARDRA was evaluated for the identification of species within the genus Streptococcus: A total of 148 type and reference strains of pyogenic, oral, and group D streptococci were examined in order to construct a database for identification of streptococci. The amplified product was a single band approximately 4500 bp long. This amplicon was digested separately with three (HhaI, MboII, and Sau3A) restriction endonucleases. Respectively, 27, 26, and 28 major patterns were observed after HhaI, MboII, and Sau3A restrictions. Streptococcal strains belonging to different species had different patterns or different combination of patterns. An identification system based upon a combination of the three restriction patterns in a single database was then proposed. rrn-ARDRA was successfully applied to 11 clinical isolates whose identification to the species level was difficult to obtain by phenotypic analysis. Using a database of well-characterized strains, rrn-ARDRA is a powerful method for the identification of streptococcal isolates.