Seroprevalence study of anti diphtheria antibodies in two age-groups of Romanian adults.

BACKGROUND: Diphtheria represents a serious infectious disease with high epidemic potential. It is a vaccine preventable disease (a minimum vaccine coverage of 95% for children of 1 year and 90% in adults could prevent the disease). Diphtheria vaccination is included in the National Immunization Program (NIP). Complete vaccination for children consists in DTaP (diphtheria toxoid, tetanus toxoid and acellular pertussis vaccine) vaccine administration from the age of 2 months until 4 years and dT vaccine (tetanus toxoid and a reduced dose of diphtheria toxoid) at 14 years old. The aim of this paper was to highlight the protection against diphtheria of an age segment of the Romanian adult population (20 to 39 years old) using a seroprevalence study. MATERIALS AND METHODS: The Romanian subjects were selected from two age groups: 20-29 years (n = 219) and 30-39 years (n = 229), representative for all counties of Romania. The commercial kit Anti-Diphtheria Toxoid ELISA (IgG) (EUROIMMUN) was used to detect the antibodies of IgG class against diphtheria toxoid in the sera obtained from our subjects. RESULTS: We detected a 56.6% rate of positive sera (> 0.1 IU/ml--protection level) for the 20-29 age group and 31.7% positivity for the 30-39 age group. These data show a low protection level against diphtheria of the Romanian adult population, which decreases with age. The serologic data on preventable vaccine diseases are useful in order to evaluate the success of the immunization programs.

Methodology optimization and diversification for the investigation of virulence potential in Haemophilus influenzae clinical strains.

Ten Haemophilus influenzae strains were isolated from patients aged between 1.6 - 24 years, with various diagnoses (acute meningitis, acute upper respiratory infection, otitis media and acute sinusitis). Identification was based on phenotypic and molecular characteristics; antibiotic susceptibility testing was performed by diffusion method according to CLSI standards 2011 for seven antibiotics. The results of molecular testing showed that all the studied strains produced an amplicon of 1000 bp with ompP2 primers indicating that all strains were H. influenzae. For six strains, the PCR amplicon obtained with bexA specific primers, proving that the strains were capsulated. The results of phenotypic testing showed that four strains were ampicillin nonsusceptible and (beta-lactamase-positive. The virulence potential of H. influenzae clinical strains was investigated by phenotypic methods, including the assessment of the soluble virulence factors on specific media containing the biochemical substratum for the investigated enzymatic factor, as well as the adherence and invasion capacity to HeLa cells monolayer using Cravioto modified method. The studied strains exhibited mainly a diffuse adherence pattern and different adherence indexes. Interestingly, two strains isolated from the same pacient (blood and CSF) showed a different degree of invasiveness, the strain isolated from blood being 20 times more invasive than the one isolated from CSF.

[Macrolides, lincosamides, streptogramines (MLS): mechanisms of action and resistance]. / Macrolide, lincozamide, streptogramine (MLS): mod de actiune si mecanisme de rezistenta.

Macrolides, lincosamides and streptogramines are distinct antibiotic (AB) families, with different chemical structure, but with similar antibacterial spectre and mechanisms. Macrolides are natural products of secondary metabolism of several species of actynomyces; they represent a group of compounds with a lactonic ring of variable dimensions (12-22 atoms of C) that can bind, by means of glycosidic bonds, sacharridic and/or amino-sacharridic structures. Most of the MLS antibiotics are bacteriostatic. Their mechanisms consist in inhibiting protein synthesis. the target being 50 S subunit of the bacterial ribosome, the binding sites being different for the different MLS classes. Erythromycin (E) was introduced in therapy in 1952; quickly, several bacterial genera started developing resistance to E. Strains resistant to E were as well resistant to all macrolides and other antibiotics with different structures--lincosamides and streptogramines B--resistance phenotype called MLSB. The main molecular mechanisms for bacterial resistance to MLS are: (1) Target modification, coded by erm genes (>12 classes). In Gram-positive cocii MLSB resistance, regardless of erm gene, can be: inducible (i MLSB)--when the presence of the inductor AB is necessary for methylation enzyme production; constitutive (c MLSB)--when the methylation enzyme is continuously produced Distinction between iMLSB and cMLSB can be easily appreciated based on the phenotypic expression of bacteria. In streptococci--all MLSB antibiotics can act as methylase inductors. (2) The decrease of AB intracellular concentration by active efflux, coded by mef genes--also called M resistance phenotype, low level resistance (LLR). (3) AB inactivation (enzymatic modification of AB); there are different resistance phenotypes: MLSB +SA and L phenotype (in staphyilococci) or SA4 phenotype and L phenotype (in enterococci).

[Study of antibiotic resistance of enterococci strains received in 2009 in the National Reference Centre for Enterococci]. / Studiul rezistentei la antibiotice a tulpinilor de enterococi primite în anul 2009 la nivelul centrului national de Referinta pentru Enterococi.

UNLABELLED: Enterococci are a frequent cause of a variety of human infections, the most common being urinary tract infection followed by bacteraemia, meningitis and endocarditis from hospitalized patients. OBJECTIVE: The study and monitoring of antibiotic resistance of enterococci strains isolated and confirmed in 2009. MATERIALS AND METHODS: Identification of 30 strains received in 2009 was based on phenotypic characteristics (microscopy, culture and biochemical characters); antibiotic susceptibility testing was performed according to CLSI standards 2009 by diffusion test and MIC by agar dilution and E-test. RESULT: The strains belonged to the following species: E. faecalis (18), E. faecium (6), E. gallinarum (5), and E. durans (1), and were isolated from: blood cultures, urine, pus-wound, cerebrospinal fluid (CSF) and catheter. Antibiotic susceptibility testing showed susceptibility to different antibiotics depending on the species and also on the type of clinical samples. CONCLUSIONS: Due to the small number of strains, no resistance phenotypes could be determined. As emergence of antibiotic resistant enterococci is well known, continuous surveillance of antibiotic susceptibility and molecular study of this issue are required.

[Study of Streptococcus pneumoniae strains resistant to antibiotics analyzed at "Cantacuzino" NIRDMI during 2006-2008]. / Studiul tulpinilor de Streptococcus pneumoniae rezistente la antibiotice analizate la INCDMI "Cantacuzino" în perioada 2006-2008.

OBJECTIVE: The aim of the this study was the analysis of the resistance to antibiotics of Streptococcus pneumoniae isolated in last years. METHODS: 328 S. pneumoniae strains, coming from blood, CSF tracheal aspirate (TA), or sputum, pleural fluid (PL) and other samples (ear and sinus fluid) isolated in 2006-2008, were analyzed at INCDMI "Cantacuzino", National Reference Center for Streptococcus pneumoniae. Strains were tested for susceptibility to by agar diution method (minimal inhibitory concentration-MIC) to the following antibiotics: penicillin (Pc), erythromycin (Em), cephalothin (Kf). cefuroxim (Cxm), cefotaxim (Ctx), trimethoprim/sulfamethoxazol (Sxt), ofloxacin (Ojx), amoxicillin (Amx). tetracycline (Te), cloramphenicol (Cm), vancomycin (Va). RESULTS: The analysis of the results was done according to CLSI 2009. Pneumococci strains isolated from blood, CSF, TA or sputum and PL showed lower resistance level to antibiotics (38.8% Pc, 9.3% Cxm. 4.1% Ctx, 2.7% Amx. 24% Em, 2.4% Ofx, 68% Sxt) against those isolated from ear ans sinus fluid which revealed high levels of resistance (70% Pc, 11.2 % Cxm, 5.9 % Ctx, 3.4% Amx, 58.4 % Em. 3.8% Ofx, 73% Sxt). Strains resistant to penicillin, isolated from blood and CSF revealed the following aspects: 17% low level of resistance and 11 % high level of resistance. CONCLUSIONS. The most efficient antibiotics were Ctx, Amx and Oft. A continuous surveillance of pneumococci strains resistant to antibiotics is needed, as well as the use of an pneumococcal efficient vaccine.

[Trends of antimicrobial resistance in microbial strains isolated from invasive infections in Romania. Experience of participation in EARSS (European Antimicrobial Resistance Surveillance System), 2002-2008]. / Tendintele rezistentei la antibiotice a izolatelor din infectii invazive în România. Experienta participarii în Sistemul European de Supraveghere a Rezistentei la Antibiotice (EARSS) în perioada 2002-2008.

EARSS (European Antimicrobial Resistance Surveillance System) is the biggest antimicrobial resistance surveillance project in the world financed from public finds, aiming to provide validated and comparable official data on antimicrobial resistance of invasive microbial strains (isolated from blood and CSF), belonging to 6 indicator bacterial species, i.e.: S. aureus, E. coli, E. faecium/faecalis, Str. pneumoniae, Ps. aeruginosa, K. pneumoniae. Romania reported data to EARSS since 2002 so far. Though the number of participating laboratories increased progressively from 12 to 35, the number of hospitals which reported for EARSS. as the number of strains included in the data base remained steady and relatively low. This issue is related to the particular position of Romania in the European context, in respect of the very low number of blood cultures performed in hospitals. Our paper is presenting the trends of antimicrobial resistance in the indicator strains in the 2002-2008 interval. During the 2002-2008 interval, Romania reported to EARSS a total number of 1276 bacterial strains, distributed by species as follows: 513 S aureus, 369 E. coli, 128 Streptococcus pneumoniae, 127 Enterococcus spp.. 71 Klebsiella pneumoniae, 68 Pseudomonas aeruginosa. Klebsiella pneumoniae and Pseudomonas aeruginosa were reported, according to the EARSS protocol, only for the 2005-2008 interval. It is difficult to describe trends, specially in Enterococcus, Streptococcus pneumonaie and the 2 species collected only since 2005, because of the low number of isolates, but there are several results that are supporting us to claim that antimicrobial resistance in invasive isolates is a real problem in Romanian hospitals, like in other Central, Southern and South Eastern European countries: more than 25% of S. aureus strains resistant to methicilline, with more than 50% in some years, high aminoglycozides resistance in more than 70-80% of Enterococcus faecium invasive strains, more than 80% of strains resistant to 3rd generation cephalosporines etc.

Group B streptococcus colonization of Romanian women: phenotypic traits of isolates from vaginal swabs.

In the attempt to enrich the local contemporary laboratory data regarding the group B streptococcus (GBS) colonization, isolates obtained from the vaginal swab cultures were characterized for their serotype distribution and antibiotic susceptibility. The 100 GBS isolates analyzed were collected during a four-month period of year 2009 from women screened in ambulatory for vaginal carriage of GBS. The GBS isolates were classified based on their capsular polysaccharide structures using commercially available antisera. Susceptibility to penicillin, ampicillin, erithromycin, clindamycin, tetracycline, ofloxacin, and chloramphenicol was initially tested using antibiotic disk diffusion technique according to CLSI guidelines. Minimum inhibitory concentrations of erythromycin and tetracycline for the isolates with reduced susceptibility were evaluated according to the CLSI criteria and macrolide-lincosamide-streptogramin B (MLSB) resistance was investigated by a double-disk test with erythromycin and clindamycin disks. All the GBS isolates were serotypeable. Their distribution comprised six different serotypes of which serotypes II (26%), III (26%), and Ia (19%) prevailed and no serotype VI, VII, and VIII isolates were found. Overall, the GBS isolates were fully susceptible to penicillin and ampicillin, but the rates of susceptibility to the other antimicrobial agents tested were decreased, ranging from 87% for chloramphenicol to 5% for tetracycline. Reduced susceptibility to clindamycin and erythromycin was detected in 18% and 19% of isolates, respectively. For the latter, 84% displayed a constitutive MLSB phenotype, 11% had an inducible MLSB phenotype, and M phenotype was expressed by 5% of them. Erythromycin-resistant GBS isolates displayed concurrently resistance to at least one more antibiotic. In conclusion, according to our study the most frequent GBS serotypes isolated from the vaginal microflora were II and III, followed by serotype Ia. While the GBS isolates remain susceptible to beta-lactams, resistance to alternative antimicrobial drugs such as erythromycin and clindamycin seems to be an increasing concern for our region. Further phenotypic and genotypic studies are required to identify specific aspects of GBS strains colonizing or infecting the local population.

Correct complete denture rehabilitation, a chance for recovering abused tissues.

The prosthodontic treatment must provide for the edentulous patients bio-functional prosthetic restorations, bio-prophylactic for the surrounding tissues. In this aim, an edentulous patient must be submitted to a methodical clinical examination in order to establish the quality of hard and soft tissues, which will indicate the degree of difficulty of the prosthetic treatment. Additional investigation as a microbiologic examination and cephalometric radiographs can be useful in a modern investigation. In our daily practice, we are rarely confronted with a normal morphology of the denture bearing oral structures. The problem of managing abused tissues in a patient with morphologic abnormalities due to faulty prostheses is sometimes difficult to solve. Preventing the deterioration of oral status must be a condition in providing a chance for the success of the following rehabilitations, mainly in the situation when the complete edentulousness succeeds in a young or middle age patient.

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.

Molecular characterization of invasive and non-invasive Streptococcus pyogenes isolates from Romania.

In 2002, the Romanian National Reference Laboratory was invited to join the Strep-EURO project to study invasive Streptococcus pyogenes infections. During 2003 and 2004, a total of 33 isolates recovered from invasive disease were received from eight Romanian counties. For comparison, 102 isolates from non-invasive disease, as well as a collection of 12 old invasive strains (isolated between 1967 and 1980) were included. All isolates were characterized by several methods: T and emm typing, presence of the fibronectin-binding protein F1 gene (prtF1), serum opacity factor (sof), and superantigen (SAg) genes (speA, speB, speC, speF, speG, speH, ssa and smeZ). The recent invasive isolates exhibited 19 emm-types, of which emm1, emm81, emm76, emm49 and emm78 covered 57 % of the strains. Furthermore, multilocus sequence typing analysis revealed nine new sequence types, corresponding to emm types 1, 12, 49, 81, 92, 100, 106 and 119. The non-invasive isolates comprised 24 different emm types with a predominance of emm1 and 12; the old invasive strains were of eight emm types, of which four were unique for this group. All isolates harboured speB and speF; smeZ was detected in all invasive strains, except for the emm49 and emm81 isolates. The majority of isolates from carriers, and patients with pharyngitis were prtF1 positive, most of these (14 strains) being emm12. High tetracycline resistance rates were noted among both invasive and control isolates (54 % and 35 %, respectively), whereas macrolide resistance rates were low (3 % and 5 %, respectively). Active and continuing surveillance is required to provide an accurate assessment of the disease burden and to provide epidemiological data on the character of isolates in Romania.

[Involvement of soluble mediators of inflammation in the pathogenic agent interaction--immune system in acute bacterial meningitis]. / Implicarea mediatorilor solubili ai inflamatiei în interactia agent patogen--sistem imun din cadrul meningitelor infectioase.

Acute Bacterial Meningitis is a medical emergency, which warrants early diagnosis and aggressive therapy, which in most cases must be initiated as an "empirical" treatment. Such an approach needs permanent epidemiological surveillance due to the major variability of the etiological agents depending upon time, geographical areas and demographic characteristics of the population. A program for the surveillance of meningitis is in progress in Romania, but the available clinical inbformation is incomplete and not well documented by paraclinical data, poorly reflecting the real incidence of the disease. The specific anatomic localization of the disease has major influences on the antiinfectious immune response. Inflammation is involved in the disease pathogenesis, especially in promotion and evolution of neurological sequelae (neuronal demyelinisation and degeneration) even in case of pathogen clearance following antimicrobial therapy. Activation of the immune response in a immunologically "privileged "region can lead to the break of tolerance and induction of autoimmunity (neuronal degenerescence). On the other hand, an efficient immune response is necessary for the clearance of pathogenic agents. A detailed investigation of the interaction between pathogenic agents and the immune system in relation to the particular meningeal localization and also a study on the involvement of soluble mediators of inflammation (cytokines, chemokines) in the pathogenesis of meningitis might prove useful for differential diagnosis (viral or "aseptic" meningitis) and also for elucidating the mechanisms which that underlie the disease pathogenesis/neurological complications.

Whooping cough in South-East Romania: a 1-year study.

The incidence of whooping cough in Romania is substantially underestimated, and, as noted by the health authorities, this is mostly due to the lack of both awareness and biological diagnosis. We conducted a 1-year study in Bucharest in order to assess the circulation of Bordetella pertussis, the main etiological agent of whooping cough. Fifty-one subjects suspected of whooping cough were enrolled. Culture, real-time PCR, and enzyme-linked immunosorbent assay were used for laboratory diagnosis. Whooping cough patients (63%) were distributed among all age groups, and most were unvaccinated, incompletely vaccinated, or had been vaccinated more than 5 years previously. Bordetella holmesii DNA was detected in 22% of the bordetellosis cases; these patients included adults; teenagers; and, surprisingly, young children. B. pertussis isolates were similar to the clinical isolates currently circulating elsewhere in Europe. One isolate does not express pertactin, an antigen included in some acellular pertussis vaccines.

Tolerance to penicillin in Staphylococcus pyogenes isolated from skin infections.

The tolerance to penicillin of S. pyogenes has been mentioned in pharyngeal strains and associated by some authors with penicillin treatment failure in streptococcal angina. In this study we followed the presence of tolerance in S. pyogenes strains isolated from children hospitalized in the Clinic of Infantile Surgery and Clinic of Dermatology Iasi for skin infections. From 138 samples of pus examined we isolated 14 strains of S. pyogenes. In 7 strains we pointed out the tolerance to penicillin by both, beta-lactamase disc test and time-killing curve methods. We did not correlate to our cases the tolerance presence with penicillin treatment failure.