Place of International Congresses in the Diffusion of Knowledge in Infectious Diseases.

Through digital resources, physicians, microbiologists, and researchers around the world can stay up-to-date with the newest developments in their field and are therefore less dependent on medical congresses as a provider of knowledge and education. The role of the medical congress in spreading knowledge in the face of this changing environment needs to be reexamined. The result is a new paradigm that thinks about the dissemination of medical knowledge and discovery as ongoing conversations between professionals and their extended networks, rather than activities that happen only during the congress. Even though the tools we use to deliver information and knowledge are rapidly evolving, there is confidence in the lasting value of meetings for medical professionals. Medical congresses are environments uniquely conducive to generating new ideas and solutions to problems. As organizers explore new ways of sharing knowledge globally, it is crucial that the high quality of medical congresses be maintained.

Field evaluation of two point-of-care tests for syphilis among men who have sex with men, Verona, Italy.

OBJECTIVES: The incidence of HIV and syphilis among men who have sex with men (MSM) in Europe has recently increased. Rapid point-of-care tests (POCTs) for syphilis can improve access to screening. The purpose of this study was to evaluate the performance of two syphilis POCTs compared with laboratory tests among MSM. METHODS: The study was undertaken in Verona, Italy. Asymptomatic MSM, potentially exposed to syphilis, were enrolled prospectively. The POCTs evaluated were SD Bioline Syphilis 3.0 and Chembio DPP Syphilis Screen & Confirm Assay on both serum and fingerprick blood. The results of the POCTs were read by the naked eye by two independent readers and their concordance assessed. RESULTS: A total of 289 MSM were enrolled in the study. Based on laboratory tests, 35 MSM (12.1%) were TPPA-positive alone and 16 (5.5%) were both Treponema pallidum particle agglutination test (TPPA) and rapid plasma reagin (RPR)-positive. The specificities of both POCTs were above 99% on both serum and fingerstick blood specimens, while sensitivities varied considerably. The sensitivity of the SD Bioline test was lower on fingerprick blood (51.4% and 54.3%, readers 1 and 2, respectively) compared with that on serum (80.0% and 82.9%). In contrast, the Chembio test exhibited similar sensitivity values for serum and fingerprick samples (57.7% and 64.0% on serum vs 65.4% and 69.2% on fingerprick for the treponemal component; 63.6% on both samples by both readers for the non-treponemal component). The positive predictive value ranged between 100% and 93.9% for the treponemal component of both syphilis POCTs, but was lower (76.3%-100%)%) for the non-treponemal component of the Chembio POCT. The negative predictive value surpassed 90% for both tests on both samples. The agreement between readers was very high (>99%). CONCLUSION: The diagnostic performance of the syphilis POCTs was lower than expected; however, considering the prevalence of syphilis among MSM, POCTs should be recommended to improve syphilis detection among MSM.

The importance of understanding the human-animal interface : from early hominins to global citizens.

The complex relationships between the human and animal species have never ceased to evolve since the emergence of the human species and have resulted in a human-animal interface that has promoted the cross-species transmission, emergence and eventual evolution of a plethora of infectious pathogens. Remarkably, most of the characteristics of the human-animal interface-as we know it today-have been established long before the end of our species pre-historical development took place, to be relentlessly shaped throughout the history of our species. More recently, changes affecting the modern human population worldwide as well as their dramatic impact on the global environment have taken domestication, agriculture, urbanization, industrialization, and colonization to unprecedented levels. This has created a unique global multi-faceted human-animal interface, associated with a major epidemiological transition that is accompanied by an unexpected rise of new and emerging infectious diseases. Importantly, these developments are largely paralleled by medical, technological, and scientific progress, continuously spurred by our never-ending combat against pathogens. The human-animal interface has most likely contributed significantly to the evolutionary shaping and historical development of our species. Investment in a better understanding of this human-animal interface will offer humankind a future head-start in the never-ending battle against infectious diseases.

Co-infection by two linezolid-resistant coagulase-negative staphylococci with two different resistance determinants.

Linezolid resistance among Gram-positive pathogens is being reported with increasing frequency. We examined 14 linezolid-resistant coagulase-negative staphylococci (CoNS) isolated from blood cultures obtained from patients admitted to the Intensive Care Unit of Vicenza General Hospital, Italy. The species identification yielded 10 Staphylococcus epidermidis, 3 Staphylococcus hominis, and 1 Staphylococcus capitis. Minimal inhibitory concentrations of linezolid ranged between 16 and 32 mg/l. By sequencing domain V of the 23S rRNA gene, 4 isolates were found to harbour a G2576T mutation and 10 isolates a G2447T mutation. None of the strains under study presented either the cfr gene or cardinal mutations in the L3, L4, or L22 riboproteins. In this clinical collection of linezolid-resistant CoNS the G2447T mutation was dominantly associated with S. epidermidis, while the G2576T mutation was found in other CoNS species. Two different CoNS species endowed with either mutation were isolated from 2 patients.

Pilot implementation of the e-Bug Project in Italy.

A pilot of the e-Bug programme has been performed in Italy. Three thousand copies of e-Bug packs have been printed and distributed to primary and first-level secondary schools in three north-eastern regions of the country (Veneto, Trentino Alto Adige and Friuli Venezia Giulia). Following the programme design in the pack, lessons based on the e-Bug resources were delivered for 1 week in a primary school in Verona. This practical approach has been adopted by other schools in these regions, in the cities of Trieste and Udine. Excellent feedback on the e-Bug packs has been received from both students and teachers. The National Italian Institute of Health has requested a further 800 primary and 800 secondary e-Bug packs for distribution to different regions.

Supranational surveillance of antimicrobial resistance: The legacy of the last decade and proposals for the future.

Antimicrobial resistance among clinically important bacteria is widely acknowledged as a major global public health threat. A decade ago, several supranational surveillance initiatives were introduced. Few of them are still ongoing, and only one features an interactive database in the public domain. No public surveillance system monitors resistance trends among non-invasive isolates on a supranational level. Although the relevance of measuring antimicrobial resistance in invasive isolates is undisputable and there is a large consensus on sampling techniques for these isolates, surveillance systems monitoring invasive infections will only have low sensitivity for early detection of emerging resistance trends, also missing an important opportunity for intervention. Surveillance of resistance patterns should ideally include characterization of important clones involved in the dissemination of resistance. This review also emphasizes important methodological issues to be considered whenever performing surveillance, and provides general recommendations applicable to surveillance at all levels.

Molecular characterization of carbapenem-resistant Klebsiella pneumoniae ST14 and ST512 causing bloodstream infections in ICU and surgery wards of a tertiary university hospital of Verona (northern Italy): co-production of KPC-3, OXA-48, and CTX-M-15 ß-lactamases.

Klebsiella pneumoniae strain is an important opportunistic pathogen that causes severe nosocomial infections. In the present study a molecular characterization of carbapenem resistant K. pneumoniae, isolated from blood samples of hospitalized patients of Verona University Hospital, was performed. The simultaneous presence of SHV-1/CTX-M-15/KPC-3 and SHV-1/CTX-M-15/OXA-48 serin-ß-lactamases was ascertained in the 89% and 11% of K. pneumoniae ST512 and K. pneumoniae ST14, respectively. Molecular characterization of bla genes showed that blaKPC-3 was found in Tn4401a transposon with the tnpR, tnpA, ISKpn6, and ISKpn7 mobile elements whereas blaCTX-M-15 was detected downstream ISEcp1 genetic element. A class 1 integron with a gene cassette of 780 bp corresponding to aadA2 gene was identified in 33 K. pneumoniae ST512 isolates.

Exploring the wound healing, anti-inflammatory, anti-pathogenic and proteomic effects of lactic acid bacteria on keratinocytes.

The topical application of lactic acid bacteria (LAB) is recognized as a useful approach to improve skin health. This work aims to characterize by a multidisciplinary approach, the wound healing, anti-inflammatory, anti-pathogens and proteomic effects of six LAB lysates, belonging to the genus Lactobacillus. Our results demonstrated that the lysates of tested LAB stimulated the proliferation of keratinocytes, and that L. plantarum SGL 07 and L. salivarius SGL 19 accelerated the re-epithelization by inducing keratinocyte migration. The bacterial lysates also reduced the secretion of specific pro-inflammatory mediators from keratinocytes. Furthermore, viable L. salivarius SGL 19 and L. fermentum SGL 10 had anti-pathogenic effects against S. aureus and S. pyogenes, while L. brevis SGL 12 and L. paracasei SGL 04 inhibited S. aureus and S. pyogenes, respectively. The tested lactobacilli lysates also induced specific proteome modulation of the exposed keratinocytes, involving dysregulation of proteins (such as interleukin enhancer-binding factor 2 and ATP-dependent RNA helicase) and pathways (such as cytokine, NF-kB, Hedgehog, and RUNX signaling) associated with their specific wound healing and anti-inflammatory effects. This study indicates the different potential of selected lactobacilli, suggesting that they may be successfully used in the future together with conventional therapies to bring relief from skin disorders.

Outbreak of Saprochaete clavata Sepsis in Hematology Patients: Combined Use of MALDI-TOF and Sequencing Strategy to Identify and Correlate the Episodes.

INTRODUCTION: New fungal species are increasingly reported in immunocompromised patients. Saprochaete clavata (S. clavata), an ascomycetous fungus formerly called Geotrichum clavatum, is intrinsically resistant to echinocandins and is often misidentified. OBJECTIVE: We describe a cluster of seven S. clavata infections in hospitalized hematology patients who developed this rare fungemia within a span of 11 months. Three of the seven patients died. Identification of the isolates was determined only with the Saramis database of VitekMS system and sequencing of the internal transcribed spacer (ITS) region. Clonal relatedness of the isolates was determined by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF) analysis; clonal correlation between the strains was investigated by means of phylogenetic analysis, based on single-nucleotide variants (SNPs). Clinical presentation, 1-3 ß-D-glucan (BG) and galactomannan (GM) antigen results and analysis of possible sources of contamination are also described with a prospective case-control study of the outbreak. RESULTS: MALDI-TOF MS-Vitek (bioMerieux, Marcy l'Etoile, France) failed to identify the six isolates, while SARAMIS (bioMerieux, Marcy l'Etoile, France) identified the isolates as S. clavata. Initially, Vitek 2 identified the strains as Geotrichum capitatum in two of the seven cases. Molecular identification gave 99% homology with S. clavata. BG was positive in three out of six patients (range 159 to >523 pg/ml), GM results were always negative. All the isolates were resistant to echinocandins (anidulafungin, micafungin, and caspofungin) and Fluconazole, but susceptible to Flucytosine and Voriconazole. One isolate showed acquired resistance to Flucytosine and Amphotericin B during treatment. Both the correlation-based dendrograms obtained by MALDI-TOF MS (Bruker Daltonics) and MS-Vitek not only clustered six of the seven bloodstream infection (BSI) isolates in the same group, but also showed their strong relatedness. Phylogenetic analysis using SNPrelate showed that the seven samples recorded during the investigation period clustered together. We observed a split between one case and the remainder with a node supported by a z-score of 2.3 (p-value = 0.021) and 16 mutations unique to each branch. CONCLUSION: The use of proteomics for identification and evaluation of strain clonality in outbreaks of rare pathogens is a promising alternative to laborious and time-consuming molecular methods, even if molecular whole-genome sequencing (WGS) typing will still remain the reference method for rare emergent pathogens.

Unusual resistance patterns in macrolide-resistant Streptococcus pyogenes harbouring erm(A).

BACKGROUND: We identified erm(A)-harbouring Streptococcus pyogenes that expressed three variant phenotypes: (1) low-level resistance to erythromycin (MICs 1-4 mg/L) but high azithromycin MICs in absolute terms (16-64 mg/L; n=6); (2) same as (1) but with a high clindamycin MIC (256 mg/L; n=1); and (3) high-level constitutive MLS (cMLS) resistance (n=1). Here we analysed the genetic basis of these novel phenotypes. METHODS: The presence of erm(A) and the absence of macrolide/lincosamide resistance genes erm(B), mef and cfr were confirmed by PCR. erm(A), 23S rRNA, L4 and L22 genes were sequenced. Mutant erm(A) genes were cloned and electrotransformed into the macrolide-susceptible Escherichia coli AG100A. Clonality was determined by emm typing and PFGE. Effects of the identified mutations on free energy changes (DeltaG) and putative configurations of the leader sequence were studied in silico. RESULTS: Point mutations (G98A, A137C, C140T and G205A) were observed in the erm(A) regulatory region of all eight erm(A)-harbouring S. pyogenes. Five and two isolates belonged to emm77 and emm89 clones, respectively, and one isolate was an emm1. E. coli transformed with mutant erm(A) harbouring G98A, A137C or C140T mutations (phenotypes 1 and 2) did not express high-level azithromycin or clindamycin resistance. However, cMLS resistance was clearly observed in transformants with erm(A) harbouring both A137C and G205A mutations (phenotype 3). In silico analysis showed that DeltaG was minor except for the G205A mutation. Secondary structure predictions further showed that the A137C and G205A mutations together abolished the hairpin sequestering the ribosome-binding and initiation sites of the erm(A) gene, explaining the cMLS phenotype 3. CONCLUSIONS: We report point mutations in the erm(A) regulatory region leading to constitutive methylase expression and the presence of additional, as yet unidentified mechanisms mediating high-level azithromycin and clindamycin resistance in erm(A)-harbouring S. pyogenes.

Investigating the Proteomic Profile of HT-29 Colon Cancer Cells After Lactobacillus kefiri SGL 13 Exposure Using the SWATH Method.

Despite some studies revealed that kefir acts on different cancers, such as colorectal cancer, the proteomic changes that occur in the colon cancer cells remain to be explored. In this study, the proteomic analysis was combined with determination of kefir characteristics (e.g., adhesion capacity, gastrointestinal and antibiotic resistances), in order to confirm its use as a probiotic. Therefore, a label-free strategy based on SWATH-MS was applied to investigate the proteomic profile of HT-29 cells after exposure for 24 h to a specific strain of Lactobacillus kefiri named SGL 13. We identified a total of 60 differentially expressed proteins in HT-29 cells, among which most are located into the extracellular exosome, playing important/crucial roles in translation and cell adhesion, as indicated by the enrichment analysis. The eIF2 and retinoid X receptor activation pathways appeared to be correlated with the anti-tumoral effect of SGL 13. Immunoblot analysis showed an increase in Bax and a decrease in caspase 3 and mutant p53, and ELISA assay revealed inhibition of IL-8 secretion from HT-29 cells stimulated with LPS upon SGL 13 treatment, suggesting pro-apoptotic and anti-inflammatory properties of kefir. In conclusion, the results of this study, the first of its kind using co-culture of kefir and colon cancer cells, demonstrate that L. kefiri SGL 13 possesses probiotic potency and contribute to elucidate the molecular mechanisms involved in the L. kefiri-colon cancer cell interactions.

Hospital antibiotic management in north-eastern Italy--results of the ABS maturity survey of the ABS International group.

BACKGROUND: The ABS International group conducted a survey to estimate the prevalence and characteristics of country-specific hospital antibiotic management programs. This paper summarizes the results for the north-eastern area of Italy. METHODS: The survey was conducted in January and February 2008. A questionnaire with items related to hospital antibiotic management was sent to the medical administrators of two regions, namely Veneto and Trentino Alto Adige, which are further subdivided into smaller administrative areas. MAIN FINDINGS: Ten out of 82 questionnaires were returned and nine were evaluable. The mean total score for all items in the areas under consideration was 3.69 (median: 3.58; range 2.58-4.50). Top values and high mean scores were achieved for diagnostics and control of antibiotic consumption. Organizational structures such as well defined antibiotic-related roles and communication tools are missing to a large degree. CONCLUSIONS: The findings of this questionnaire-based survey underline the considerable potential for further improvement of antibiotic stewardship in north-eastern Italy.

Non-compliance with antibiotic therapy for acute community infections: a global survey.

A global patient survey of non-compliance with antibiotic therapy for acute community infections included 4514 adult respondents (aged 18-99 years) in 11 countries. Admitted non-compliance (ANC) was reported in 912/4088 (22.3%) of cases but varied widely between countries. Multivariate analysis identified five independent variables associated with ANC: country, daily dosage regimen, age, attitudes to doctors and attitudes to antibiotics. ANC ranged from 44.0% in China to 9.9% in The Netherlands, and from 14.9% in those prescribed once-daily regimens to 27.0% for three or more daily doses. There was a negative correlation between ANC and age. Analysis of the attitudes to doctors confirmed previous findings that involving the patient in the management of their infection can improve overall compliance. The study identified seven key attitudes to antibiotic use with the potential to improve compliance. However, there was a poor understanding in 10 of the 11 countries of how non-compliance can increase the potential for resistance development. Segmentation and cluster analysis identified four psychographic profiles influencing compliant behaviour, which varied across countries. The global picture of antibiotic non-compliance and psychographic profiling should help identify areas for targeted, country-specific patient educational programmes as well as those areas where physicians can improve their interaction with their patients.

A global survey of antibiotic leftovers in the outpatient setting.

We performed a global survey of outpatients who had taken self-administered antibiotics within the last 12 months in order to identify factors that lead to possession of leftover antibiotics in the community. The study included 4,514 subjects aged 18-99 years. Of 4,192 respondents not currently taking antibiotics, 53.7% admitted having leftover antibiotics, of whom 77.0% saved them, 4.6% gave them away and 18.4% threw them away. Living in a country where antibiotics are dispensed in fixed packs rather than exact numbers of pills as well as believing that leftover antibiotics can be saved and used again were the strongest predictors for possession of leftovers. There was also a marked detrimental effect of lack of information from the doctor and/or pharmacist. This investigation suggests that dispensing of antibiotics in exact numbers of doses should be recommended in addition to the development of relevant information campaigns addressing patients' false beliefs about leftovers and the provision of basic information about the importance of completing antibiotic treatment.

Metallo-beta-lactamases as emerging resistance determinants in Gram-negative pathogens: open issues.

The rapid spread of acquired metallo-beta-lactamases (MBLs) among major Gram-negative pathogens is a matter of particular concern worldwide and primarily in Europe, one of first continents where the emergence of acquired MBLs has been reported and possibly the geographical area where the increasing diversity of these enzymes and the number of bacterial species affected are most impressive. This spread has not been paralleled by accuracy/standardisation of detection methods, completeness of epidemiological knowledge or a clear understanding of what MBL production entails in terms of clinical impact, hospital infection control and antimicrobial chemotherapy. A number of European experts in the field met to review the current knowledge on this phenomenon, to point out open issues and to reinforce and relate to one another the existing activities set forth by research institutes, scientific societies and European Union-driven networks.

[Antibacterial activity and pharmacological features of fluoroquinolones and considerations on their use in a nosocomial setting]. / Attivita antibatterica e caratteristiche farmacologiche dei fluorochinoloni e considerazioni sul loro impiego ospedaliero.

The need for appropriate use of antimicrobials limiting the insurgence, selection and spread of bacterial resistance and preventing its diffusion to the community is increasingly widely felt. Based on some CDC expert recommendations, an antibiotic therapy should be driven by the following criteria: scientifically proven clinical evidence, high probability of being beneficial to patients, safety and tolerability, drug and regimen choice made according to the infection to be treated, and cost/efficacy ratio. In this article, we compare the microbiological and pharmacological characteristics of the three most frequently used fluoroquinolones in Italy - ciprofloxacin, levofloxacin, and moxifloxacin - highlighting the main differences and peculiar aspects for each of them. We describe the microbiological profile, providing details on bacterial resistance mechanisms, the pharmacological profile, giving details on tolerability, and the clinical profile, showing the individual therapeutic efficacy, underlining the main differences in their pharmacodynamic and pharmacokinetic aspects. The main national and international guidelines recommend ciprofloxacin and levofloxacin and, to a lesser extent, moxifloxacin, for treating nosocomial infections localised in different organs and apparatuses, including severe infections in critical patients. In addition, they represent a basic therapeutic option for treatment of community-acquired infections, such as urinary and respiratory tract infections.

Multi-drug-resistant Gram-negative bacteria causing urinary tract infections: a review.

Urinary tract infections (UTIs) are among the most frequent infectious diseases affecting humans, and represent an important public health problem with a substantial economic burden. Due to the high empiric use of antibiotics for the treatment of UTI, antibacterial resistance of Enterobacteriaceae, specifically the main uropathogens Escherichia coli and Klebsiella pneumoniae, has significantly increased worldwide. In this article the worldwide epidemiology of resistant Gram-negative bacteria causing UTIs, with a special focus on extended spectrum beta lactamase (ESBL) positive pathogens, as well as new threats such as multi-drug-resistant (MDR) clones (e.g. E. coli 131 (ST131) and K. pneumoniae ST258), are reviewed. The increased prevalence of MDR Enterobacteriaceae, limiting available treatment options for infections caused by these organisms, and the lack of new antibiotics provide good rationale for using older antibiotics, such as fosfomycin, that have been shown to retain some activity against MDR bacteria.

blaNDM-1 Carriage on IncR Plasmid in Enterobacteriaceae Strains.

Four NDM-1-producing Enterobacteriaceae strains (three Klebsiella pneumoniae and one Citrobacter koseri) were isolated between 2009 and 2011 through a nationwide surveillance for carbapenem-resistant Enterobacteriaceae in Croatia to study the molecular genetic background of blaNDM and the responsible plasmid types. Phenotypically, the clinical strains proved to be multidrug resistant. All strains remained susceptible to tigecycline and colistin. The clinical strains harbored variable antibiotic resistance determinants, notably, blaNDM-1, blaTEM-1, blaSHV-1, blaSHV-12, blaOXA-1, blaOXA-9, blaCTX-M-15, blaCMY-4, qnrB1, and aac(6')Ib-cr in different combinations. Two K. pneumoniae belonged to sequence type ST15 and one strain to ST16. As for the plasmid types, C. koseri and one of the ST15 K. pneumoniae carried IncR, and the second ST15 K. pneumoniae carried IncR and colE. The K. pneumoniae ST16 strain hosted A/C and colE plasmids. The blaNDM-1 gene was detected on conjugative high-molecular-weight plasmids, namely, A/C and IncR types. It is noteworthy that this is the first description of K. pneumoniae ST16 expressing NDM-1 in Europe. Remarkably, our study underscores the importance of the IncR plasmid as a reservoir of multidrug resistance. To the best of our knowledge, the IncR plasmid carrying blaNDM-1 in C. koseri is reported for the first time.