Probiotic strains of Lactobacillus brevis and Lactobacillus plantarum as adjunct to non-surgical periodontal therapy: 3-month results of a randomized controlled clinical trial.

OBJECTIVES: To determine if periodontitis patients benefit from treatment with Lactobacillus brevis and Lactobacillus plantarum strains, applied into periodontal pockets as gel and thereafter taken as lozenges, as an adjunct to scaling and root planing (SRP). MATERIALS AND METHODS: In a double-blind, randomized, placebo-controlled trial, 40 patients received scaling and root planing (SRP) in two sessions within 7 days. Patients then received either probiotic gel and lozenges (n = 20) or placebo (n = 20). The primary outcome variable was the number of diseased sites (DS: PD > 4 mm + BOP) at the 3-month re-evaluation. The effects of gender, age, probiotic therapy, presence of Porphyromonas gingivalis or Aggregatibacter actinomycetemcomitans, smoking, tooth being a molar and interdental location were evaluated using a multivariate multilevel logistic regression model. RESULTS: The number of DS after 3 months was similar in the test (Me = 8, IQR = 5-11) and control (Me = 5, IQR = 1-10) groups. Both groups showed substantial but equivalent improvements in periodontal parameters. The logistic regression showed higher odds for the healing of gingival bleeding (OR = 2.12, p = 0.048) and lower odds for the healing of DS (OR = 0.51; p < 0.001) in the probiotic group. CONCLUSIONS: Patients with periodontitis benefit from adjunctive use of probiotics containing L. brevis and L. plantarum in terms of reduction of gingival bleeding. However, adjunctive probiotics increase the number of persisting diseased sites with PD > 4 mm and BOP. CLINICAL RELEVANCE: The adjunctive use of probiotics containing L. brevis and L. plantarum strains in treating chronic periodontitis results in a higher number of residual diseased sites when compared with SRP + placebo; its use is therefore unfounded.

Successful control of the first OXA-48 and/or NDM carbapenemase-producing Klebsiella pneumoniae outbreak in Slovenia 2014-2016.

BACKGROUND: Carbapenemase-producing Enterobacteriaceae (CPE) occur only sporadically in Slovenia. AIM: To describe the first Slovenian carbapenemase-producing (CP) Klebsiella pneumoniae and Escherichia coli outbreak which occurred at the tertiary teaching hospital University Medical Centre Ljubljana from October 2014 to April 2015. METHODS: A CPE-positive case was defined as any patient infected or colonized with CPE. A strict definition of a contact patient was adopted. Measures to prevent cross-transmission included cohorting of all CPE carriers with strict contact precautions and assignment of dedicated healthcare workers, cohorting of all contact patients until obtaining the result of screening cultures, systematic rectal screening of contact patients, and tagging of all CPE-positive cases and their contacts. Educational campaigns on CPEs were implemented. Clinical specimens were processed using standard procedures. Pulsed-field gel electrophoresis (PFGE) was used to determine relatedness. Multi-locus sequence typing was performed on CP K. pneumoniae isolates that belonged to different pulsotypes. FINDINGS: Before the outbreak was brought under control, 40 patients were colonized or infected with OXA-48 and/or New Delhi metallo-ß-lactamase (NDM)-producing CPE; in 38 patients OXA-48 and/or NDM-producing K. pneumoniae was detected, in seven OXA-48 and/or NDM-producing E. coli was found together with K. pneumoniae, and in two patients only CP E. coli was isolated. The outbreak was oligoclonal with two major CP K. pneumoniae clusters belonging to ST437 and ST147 in epidemiologically linked patients. CONCLUSION: Initial standard control measures failed to prevent the outbreak. Once the problem had been recognized, strict infection control measures and the education of healthcare workers contributed to the successful control of the outbreak.

Hepatitis C virus infection among pregnant women in Slovenia: study on 31,849 samples obtained in four screening rounds during 1999, 2003, 2009 and 2013.

The majority of people infected with hepatitis C virus (HCV) are unaware of their infection. Assessment of the prevalence of HCV infection in the general population and in key populations at increased risk is needed for evidence-based testing policies. Our objectives were to estimate the prevalence of antibodies to HCV (anti-HCV), the prevalence of HCV viraemia (HCV RNA), and to describe HCV genotype distribution among pregnant women in Slovenia. Unlinked anonymous testing was performed on residual sera obtained from 31,849 pregnant women for routine syphilis screening during 1999, 2003, 2009, and 2013. Anti-HCV reactive specimens were tested for HCV RNA and HCV genotypes were determined. Annual prevalence of anti-HCV ranged between 0.09% (95% confidence interval (CI): 0.03­0.18) in 2009 and 0.21% (95% CI: 0.12­0.34) in 2003 and HCV RNA positivity between 0.06% (95% CI: 0.02­0.14) in 2009 and 0.14% (95% CI: 0.07­0.25) in 2003. We observed no statistically significant differences in anti-HCV or HCV RNA prevalence between age groups (<20, 20­29 and ≥30 years) in any year and no trend in time. Of 29 HCV active infections, 19 were with genotype 1 and 10 with genotype 3. HCV infection among pregnant women was rare suggesting a low burden in the Slovenian general population. Antenatal screening for HCV in Slovenia could not be recommended.

Antibiotic susceptibility profiles of oral pathogens.

Periodontitis is a bacterial disease that can be treated with systemic antibiotics. The aim of this study was to establish the antibiotic susceptibility profiles of five periodontal pathogens to six commonly used antibiotics in periodontics. A total of 247 periodontal bacterial isolates were tested for susceptibility to the six antibiotics using the Etest method. MIC(50) and MIC(90) values (minimum inhibitory concentrations for 50% and 90% of the organisms, respectively) were calculated. Both European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) breakpoints were used in the study to interpret results. ß-Lactamase production was tested when amoxicillin resistance was found. MIC(90) values of the anaerobic bacteria were all well below breakpoint values, except for three isolates of Prevotella intermedia and one isolate of Fusobacterium nucleatum that were resistant to amoxicillin (CLSI breakpoints); these isolates were ß-lactamase-positive. Two isolates of the capnophilic Aggregatibacter actinomycetemcomitans appeared to be amoxicillin-resistant but failed to show ß-lactamase activity. Comparison with a previous study from The Netherlands showed minor differences in susceptibility profiles, but the MIC(90) values of A. actinomycetemcomitans for amoxicillin, clindamycin, azithromycin and tetracycline were higher. Geographical differences in the susceptibility profiles of Porphyromonas gingivalis and A. actinomycetemcomitans between European countries were noted. Comparison of European susceptibility profiles with that of a South American country (Colombia) revealed a much higher resistance in the latter. Owing to these differences in susceptibility profiles, it is of concern to regularly perform surveillance studies on antibiotic resistance.

Dissemination of New Delhi metallo-ß-lactamase-1-producing Acinetobacter baumannii in Europe.

Multidrug-resistant and New Delhi metallo-ß-lactamase 1 (NDM-1) -producing Acinetobacter baumannii are increasingly reported. A collection of five NDM-1-positive A. baumannii isolates recovered in four European countries were analysed. Genotyping was performed by pulsed-field gel electrophoresis, multiplex PCR sequence typing, Diversilab and multilocus sequence typing. Three distinct sequence types were identified. All isolates harboured a chromosomally located bla(NDM-1) gene within a Tn125-like transposon. One isolate co-expressed another unrelated carbapenemase OXA-23. This report constitutes the first epidemiological study of NDM-1-producing A. baumannii from four countries.

Does ertepenem alter the susceptibility of Pseudomonas aeruginosa to carbapenems?

We investigated the effect of ertapenem on carbapenem susceptibility of Pseudomonas aeruginosa. Antibiotic consumption was recorded monthly in defined daily doses (DDD)/100 patient-days in the infectious diseases (ID), abdominal surgery (AS), and surgical intensive care units (SICU) of a teaching hospital from January 2005 to December 2008. Trends of decreased susceptibility of P. aeruginosa were observed in all three units. After the introduction of ertapenem, the number of P. aeruginosa isolates/ 1000 patients-days per month increased in AS and in SICU (p=0.05). The increase in carbapenem non-susceptible isolates/1000 patients-days in the same units was less significant (p=0.07 and p=0.054). Correlations between ertapenem and the carbapenem non-susceptibility for the lagtime of 1 to 6 months ahead gave no significant result. In the SICU, 30% of variability of carbapenem non-susceptibility could be predicted by the consumption of ertapenem. There is no evidence that ertapenem alters the P. aeruginosa susceptibility to carbapenems, but the relationship deserves further observation.

A case of OXA-48 carbapenemase-producing Klebsiella pneumoniae in a patient transferred to Slovenia from Libya, November 2011.

We report the first documented case of OXA-48-producing Klebsiella pneumoniae in Slovenia isolated from rectal surveillance cultures from a patient transferred from Libya. The patient was colonised with both ESBL-producing Escherichia coli and ESBL- and OXA-48-producing K. pneumoniae. Three further patients were colonised with ESBL-producing E. coli. This underscores the importance of an early warning system on European level and screening upon admission of patients transferred across borders and between healthcare systems.

Antimicrobial susceptibility, beta-lactamase and enterotoxin production in Bacillus cereus isolates from clinical and food samples.

The antimicrobial susceptibility of 30 clinical and 30 food Bacillus cereus isolates was determined. All isolates were susceptible to streptomycin, ciprofloxacin and gentamicin, 90 % of them to clindamycin and vancomycin, and 67 % to erythromycin. All isolates were resistant to amoxicillin with clavulanic acid, ampicillin, cefotaxime, ciprofloxacin, cloxacillin, cefotaxime with clavulanic acid and penicillin. The MIC values (determined by E-tests) were 48-256 mg/L for ampicillin, 0.19-1.5 mg/L for gentamicin, 0.125-1.0 mg/L for clindamycin, 0.047-4.0 mg/L for erythromycin and 1.5-16 mg/L for vancomycin. The MICs 4.6-18.75 g/L were observed for penicillin using the microdilution method. The presence of metallo-beta-lactamases was detected by E-test for 100 % of strains. Nonhemolytic diarrheal enterotoxin (NHE) was produced by 98.3 % of strains, while 31.7 % of them produced hemolytic diarrheal enterotoxin (HBL). Clinical isolates produced 10 % more HBL than food isolates. The psychrotrophic strains isolated from food samples produced NHE at 6.5 degrees C in 73 % of cases.

Pre-vaccination distribution of human papillomavirus (HPV) genotypes in women with cervical intraepithelial neoplasia grade 3 (CIN 3) lesions in Slovenia.

BACKGROUND: High-risk human papillomaviruses (HPV) are the main etiological factor of cervical cancer. Cervical intraepithelial neoplasia grade 3 (CIN 3) is the latest pre-invasive stage of cervical cancer, with an approximately 20% progression rate to invasive cervical carcinoma. OBJECTIVE: To establish the pre-vaccination distribution of HPV genotypes in Slovenian women with CIN 3 lesions, in order to assess the potential benefit of prophylactic HPV vaccination in Slovenia, and to provide baseline data for monitoring the potential replacement of HPV genotypes under selective pressure of HPV vaccines. METHODS AND RESULTS: A total of 261 cervical swabs collected from women with histologically confirmed CIN 3 lesions were analyzed using four genotyping methods: the Abbott RealTime High Risk HPV Assay, the Innogenetics INNO-LiPA HPV Genotyping Extra Test, and the in-house PGMY09/11, and CPI/CPIIg polymerase chain reaction (PCR) and sequencing. Of 261 samples, 253 (96.9%) were HPV positive. The most common HPV genotype in CIN 3 lesions in the Slovenian samples was HPV-16 (59.0%), followed by HPV-31 (7.5%), HPV-33 (7.1%), HPV-58 (5.0%), and HPV-51 (4.0%). The presence of more than one HPV genotype was detected in 49/253 (19.4%) samples. Together, HPV-16 and HPV- 18 accounted for 67.4% of CIN 3 lesions in this Slovenian population. CONCLUSION: The high proportion of CIN 3 lesions caused by HPV-16 and HPV-18 should further support the recent decision to include the prophylactic vaccination against HPV in the national vaccination program in Slovenia.

The impact of total control of antibiotic prescribing by infectious disease specialist on antibiotic consumption and cost.

We analyzed the impact on antimicrobial consumption of antimicrobial therapy totally guided by an infectious disease specialist (iD). in a teaching hospital antibiotic consumption was recorded yearly from 1998 to 2005 in three units. Antibiotic policy was introduced in 1999. in Unit A, restricted antimicrobials were prescribed after approval by the head of the unit. the iD was involved as a consultant upon request. in unit b, restricted antimicrobials were approved by an iD. All other antimicrobials were prescribed by the physicians on the ward. in unit C, all antimicrobials were prescribed by an iD. A significant decreasing trend in antibiotic consumption in defined daily doses (DDD) per patient day and per admission, and cost of antimicrobials per patient was observed in unit C, a decreasing trend in antibiotic consumption per patient was observed in unit b as well. totally iD guided antimicrobial therapy is an efficient method of antimicrobial policy.

Nationwide survey of CTX-M-type extended-spectrum beta-lactamases among Klebsiella pneumoniae isolates in Slovenian hospitals.

Among 177 extended-spectrum beta-lactamase-producing Klebsiella pneumoniae isolates collected from 11 Slovenian hospitals in 2005 and 2006, 60 (34%), from eight hospitals, harbored genes for CTX-M enzymes, with bla(CTX-M-15) detected by sequencing. These 60 isolates comprised 11 pulsed-field gel electrophoresis-defined strains, with several clusters of closely related isolates. Plasmids encoding CTX-M-15 enzyme were highly transmissible.

Distribution of human papillomavirus (HPV) genotypes in genital warts from males in Slovenia.

BACKGROUND: Genital warts (GWs) are the most frequent benign tumors in the anogenital region of both males and females. Human papillomaviruses (HPV) are etiologically associated with the development of virtually all GWs. HPV-6 and HPV-11 are the most commonly detected HPV genotypes, but at least 20 other alpha-HPV genotypes have occasionally been found in GW tissue specimens. OBJECTIVE: There is limited knowledge of GWs in Slovenia. Thus in this study we tested 55 GW tissue specimens collected from the same number of male patients using 2 different PCR protocols to obtain the first data concerning HPV and GWs in Slovenia. MATERIAL AND METHODS: 55 GW tissue specimens were tested for the presence of HPV using PGMY09/PGMY11 and CPI/CPIIg polymerase chain reaction (PCR). HPV genotypes were determined using restriction fragment length polymorphism analysis of PGMY09/11 PCR products or by sequencing of the CPI/CPIIg PCR products. In some GWs, the genotyping results were also confirmed using the Linear Array HPV Genotyping Test. RESULTS: HPV DNA was detected in all 55 tissue specimens of GWs. HPV-6 or HPV-11 was detected in 53 cases of GWs, and HPV-44 and candHPV-91 in one GW each. HPV-6 was detected approximately 4 times more frequently than HPV-11. In addition, HPV-16, HPV-31, HPV-51, HPV-53, HPV-55, candHPV-62, HPV-66, HPV-70, HPV-73, and HPV-84 were detected in some GW specimens. According to the published data, our study is the first to report the presence of candHPV-62 and candHPV-91 in GW tissue specimens. CONCLUSIONS: Our study showed that HPV can be found in virtually all GW tissue specimens obtained from male patients in Slovenia. Because HPV-6 or HPV-11 was detected in 96.4% of GWs studied, it seems that, if a quadrivalent HPV vaccine proves to be effective in males, this vaccine could prevent the great majority of incidental GWs in males in Slovenia.

Retrospective and prospective evaluation of the Amplicor HPV test for detection of 13 high-risk human papillomavirus genotypes on 862 clinical samples.

BACKGROUND: Persistent infection with a subgroup of at least 15 high-risk human papillomavirus (HPV) genotypes is considered as a necessary although insufficient etiological factor in the development of cervical carcinoma. As a consequence, HPV testing has recently become an important part of the cervical carcinoma screening and detection algorithms. AIM OF THE STUDY: To evaluate the analytical performance of a recently developed polymerase chain reaction (PCR)-based Amplicor HPV test (Roche Molecular Systems) in comparison with the Hybrid Capture 2 HPV DNA test (hc2) (Digene Corporation) for the detection of 13 high-risk HPV genotypes. Inhouse consensus PGMY09/PGMY11 and CPI/IIg PCRs targeting two different HPV genes coupled with HPV genotyping were used as an HPV internal reference standard. MATERIALS AND METHODS: In the retrospective evaluation, 550 cervical scrape specimens with previously established HPV status were included. Additionally, 312 cervical scrape specimens were tested prospectively for the presence of 13 high-risk HPV genotypes by both hc2 and Amplicor HPV test. RESULTS: In the retrospective evaluation, the Amplicor HPV test results agreed almost completely with the HPV internal reference standard results. In the prospective evaluation performed on 312 samples, the concordant Amplicor and hc2 results were obtained in 85.9% of samples tested. CONCLUSION: In our hands, the Amplicor HPV test demonstrated high analytical sensitivity and specificity. The higher analytical specificity of Amplicor in comparison to that of hc2 can be considered clinically useful. Prospective studies with clinical endpoints are urgently needed to assess the clinical utility of the higher analytical sensitivity of the Amplicor HPV test for primary HPV screening and triaging patients with ASC-US.

Treatment of chronic hepatitis C: our experience.

BACKGROUND/AIMS: While an optimal treatment of chronic hepatitis C has not yet been established, it has been demonstrated that the interferon alpha/ribavirin combination is more effective than interferon alpha monotherapy. METHODOLOGY: One hundred and forty-three patients with chronic hepatitis C received the following treatment: eighty patients an 18-month monotherapy (3-month follow-up) and sixty-three patients a 12-month combined therapy (6-month follow-up). Therapeutic efficacy and adverse effects were compared. RESULTS: In 80 patients in the monotherapy group, complete response was achieved in 49.2%. This was reduced to 27.5% three months after therapy. Significant differences were observed in HCV 3 genotype where complete response was achieved in 12 out of 14 patients (p=0.01). With the combined therapy administered to 63 patients, complete response was achieved in 54.5%. This was reduced to 43.2% after 6 months of follow-up. Among the responders or partial responders, significant differences were observed with regard to age (p=0.0047) and subtype 1b (p=0.012). Comparing the groups of naive patients and relapsers, a statistically significant difference (p=0.027) was found in therapeutic efficacy. CONCLUSIONS: In the treatment of chronic hepatitis C, combined therapy proved more effective than monotherapy. This is, however, not yet a satisfactory solution.

Human papillomavirus DNA in oral squamous cell carcinomas and normal oral mucosa.

To elucidate the putative etiologic role of human papillomaviruses (HPV) in oral carcinogenesis, a comparative study was carried out on 62 tissue specimens of oral squamous cell carcinoma (OSCC) and on 62 specimens of histologically normal oral mucosa obtained from the individuals who matched the subjects with OSCC in age, gender, localization of obtained tissue specimens, drinking and smoking habits. Internal control amplification showed that amplifiable DNA was recovered from 59/62 and 61/62 tissue samples of OSCC and normal oral mucosa, respectively. The amplification with two different HPV L1 and one HPV E6 consensus primer sets showed the presence of the HPV DNA genotypes 16, 33, 58 in 5/59 (8.4%) OSCC specimens and HPV genotypes 11, 16, 31, 68 in 4/61 (6.6%) tissue samples of normal oral mucosa tested. In the study in which a comparative examination of the presence of HPV DNA was for the first time performed on the tissue samples of the patients with OSCC and the age- and gender-matched control subjects there was no significant difference in the prevalence of HPV DNA among both study groups. Our results suggest that occasional findings of HPV DNA in OSCC tissue specimens may be the result of an incidental HPV colonization of oral mucosa, rather than of viral infection, and that HPVs play a limited role in the etiopathogenesis of the majority of OSCC.

The IgG antibody profile to various antigen regions of hepatitis C virus differs in oral fluid and serum of patients with chronic hepatitis C.

Antibodies to hepatitis C virus (HCV) can be detected not only in serum but also in oral fluid. The aim of the study was to determine IgG antibody reactivity directed to six antigen regions of HCV in oral fluid and to evaluate the significance of the antibody pattern in oral fluid compared to serum. Oral fluid and serum samples of 32 HCV viremic patients were collected to detect antibodies to six antigen regions incorporated as antigen bands into modified commercial updated third generation line immuno-assay. Compared to serum, a significantly lower cumulative antibody response and reactivity to five HCV antigens was found in oral fluid. The significantly highest prevalence of oral fluid reactivity was recorded with antigen C1 (78%), whereas in serum the most significantly frequent reactivity was detected with antigen NS3 (100%). The absence of antibody reactivity with antigen E2 was similar in both body fluids. The discrepancy in antibody pattern to HCV antigens between oral fluid and serum indicates the possible existence of local viral replication, viral mutants, viral inhibitors in oral cavity and, most probably, leakage of the muco-vascular barrier.

Low prevalence of hepatitis C virus infection among human immunodeficiency virus type 1-infected individuals from Slovenia and Croatia.

The prevalence of hepatitis C virus (HCV) infection in the population of human immunodeficiency virus 1 (HIV-1)-infected individuals from Slovenia and Croatia was determined. One hundred and sixty-six out of a total of 188 Slovenian HIV-1-infected individuals and 120 subjects who were randomly chosen out of a total 342 Croatian HIV-1 antibodies-positive individuals were tested for HCV infection. Detection of HCV antibodies was carried out by a third generation enzyme-linked immunoassay (ELISA) and the positive samples were additionally tested by a third generation immuno-blot assay. Additionally, the presence of HCV RNA was determined in all serum samples by a qualitative polymerase chain reaction (PCR). Twenty-four (14.5%) out of 166 Slovenian and 18 (15.0%) out of 120 Croatian HIV-1-infected individuals were HCV antibodies-positive. Nineteen out of 24 (79.2%) Slovenian and 13 out of 18 (72.2%) Croatian anti-HCV positive individuals were also viremic. HCV RNA was not detected in any of 244 HCV antibodies-negative/HIV-1-infected individual from both countries. A significant difference in the prevalence of HCV infection between blood (77.8% in Slovenia and 66.7% in Croatia) and sexual exposure risk groups (1.6% in Slovenia and 6.6% in Croatia) was found in both countries. In a study carried out on the highest proportion of entire population of HIV-1-infected individuals from a certain country or geographic region, Slovenia and Croatia were identified as countries with the second and third lowest prevalence of HCV infection among HIV-1/HIV-2 infected individuals worldwide.

Second-generation Hybrid capture test and Amplicor monitor test generate highly correlated hepatitis B virus DNA levels.

The performance of the Digene Hybrid Capture II HBV DNA Test HC II and the Roche Cobas Amplicor Monitor Test (Cobas-HBV) was evaluated on 252 serum samples. One hundred and seventy-three samples were HBV DNA positive and 75 HBV DNA negative by both assays. Four samples were HBV DNA positive by Cobas-HBV only. Linear regression analysis showed that the HBV DNA concentrations obtained from both assays were significantly related (n=173, r=0.976, P<0.0001). The results of the study show that Hybrid capture II and Cobas-HBV could be used equally in the management for patients with chronic HBV infection.