PURPOSE: To report a case of sympathetic ophthalmia (SO) after a severe Acanthamoeba keratitis. METHODS/DESIGN: Interventional case report. RESULTS: A 59-year-old white woman, wearing contact lenses, developed a severe Acanthamoeba keratitis in the left eye, which involved the limbus, and required 8 months of intensive antiamoeba therapy; the condition resolved leaving a painful, phthisical eye with complete corneal neovascularization. Six months later, the patient presented with pain, blurred vision, and photophobia in the right eye. Slitlamp examination of the right eye revealed granulomatous uveitis. On the suspicion of an SO, treatment with high-dose topical and oral corticosteroids and immunosuppressants was started. After 3 months, the eye is stable, with a visual acuity of 20/50, and the patient is taking prednisolone 7.5 mg per day and cyclophosphamide 50 mg per day. CONCULSIONS: Development of SO in the absence of previous trauma or surgery is rare. Our case is the first report of a clinically diagnosed SO after an episode of severe Acanthamoeba keratitis.

This study evaluated the diagnostic performances of an ELISA method and a molecular method for the detection of verotoxin in faecal samples during an outbreak of haemolytic-uraemic syndrome (HUS) occurring in Apulia, Southern Italy. Two of the 16 faecal samples were positive for verotoxin when analysed by ELISA and resulted PCR positive for stx1, stx2, eaeA and serogroup O26. The other 14 faecal samples resulted negative with both tests. The detection of verotoxin in faecal samples by ELISA is a simple, sensitive, specific and rapid method (2 hours) of considerable utility for routine clinical testing laboratories without access to more specialized diagnostic procedures.

Sixty-two multidrug resistant Salmonella enterica serovar Typhimurium strains isolated from 255 clinical strains collected in Southern Italy in 2006–2008 were characterised for antimicrobial resistance genes, pulsotype, and phage type.Most strains (83.9%) were resistant to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline (resistance pattern ACSSuT) encoded in 88.5% by the PSE-1, floR, aadA2, sul1, and tet(G) gene cluster harboured by the Salmonella Genomic Island (SGI1). In 11.5% of strains, the resistance was encoded by the InH-like integron (OXA-30-aadA1) and the catA1, sul1, and tet(B) genes. STYMXB.0061 (75%) and DT120 (84.6%) were the prevalent pulsotype and phage type identified in these strains, respectively. Five other resistance patterns were also found either in single or in a low number of isolates with TEM, dfrA12, strAB, sul2, tet(A), and tet(B) encoding for the associated ampicillin, trimethoprim, streptomycin, sulfamethoxazole, and tetracycline resistances, respectively. The pandemic DT104 clone, resistance pattern ACSSuT encoded by SGI1, has largely been identified in Italy since 1992, while strains DT120, resistance pattern ACSSuT (encoded by SGI1), have never been previously reported in Italy. In Europe, clinical S. Typhimurium strains DT120 have mainly been reported from sporadic outbreaks linked to the consumption of pork products.However, none of these strains were STYMXB.0061 and the antimicrobial resistance was not linked to SGI1.Theprevalent identification and persistence ofDT120 isolates would suggest, in Southern Italy, a phage type shifting of the pandemic DT104 clone pulsotype STYMXB.0061.Additionally, these findings raise epidemiological concern about the potential diffusion of these emerging multidrug resistant (SGI linked) DT120 strains.

Bacterial intestinal overgrowth syndrome (SIBO) treatment is based on antibiotics. Probiotics have been shown to give similar results, whilst no study is available about prebiotics. This study evaluated the addition of probiotics or prebiotics to antibiotics on SIBO symptoms in a 6-month follow-up. We enrolled 40 patients (14 males and 26 females) reporting abdominal compliant without gastrointestinal diseases/alarm symptoms. SIBO was diagnosed by the agreement of lactulose and glucose breath tests. Patients were randomly divided into two groups homogeneous for sex and age: group 1 received Rifaximin 400 mg/day for 7 days/month followed by Lactobacillus casei for 7 days more and group 2 antibiotic followed by short chain fructo-oligosaccharides. All patients recorded a questionnaire for subjective symptom evaluation according to Rome III criteria and Bristol scale for stool characters before the study and after 6 months. Statistics: Student's t and Fisher's exact tests. In group 1, a significant improvement was obtained in 5 out of 6 symptoms, whilst in group 2 in 4 out of 6 symptoms (nausea and number of bowel movements failed to improve). Despite we observed a trend of probiotics to be more effective than prebiotics, the difference in the percentage of improved symptoms was not significant (83,3% vs 66.6%; p= 0.57). Our preliminary data show a good outcome with sequential antibioticprobiotic/ prebiotic administration in patients with SIBO.

We evaluated the usefulness of a rapid immunochromatographic pneumococcal urinary antigen test (UAT) for the diagnosis of pneumonia over a period of five years. The UAT was positive in 32 (2.3%) urine samples obtained from 1414 patients. In 46 of these 1414 patients results of UAT and/or sputum/pleural fluid culture and/or blood culture and/or procalcitonin levels were available and therefore the study was concentrated on these patients. A concordance between UAT positivity and the presence of Streptococcus pneumoniae in the sputum was observed in only 4 of 46 (8.7%) patients for which both urine and sputum samples were analyzed. A discordant result (UAT positive and absence of S. pneumoniae in sputum samples) was recorded in 8 of 46 (17.4 %) patients. UAT negative results with sputum culture positive for S. pneumoniae were recorded in 28.3% of patients. In 20 patients, UAT tested positive but sputum culture was not performed. A concordance between UAT positivity and the isolation of S. pneumoniae from blood was seen in 2 of 46 patients whereas a discordant result (UAT positive and blood culture negative) was seen in 12 (26.1%) patients. A concordance between the UAT and high levels (≥2ng/ml) of procalcitonin was observed in 4 out of 46 patients, whereas a positive UAT result and a procalcitonin negative result were observed in 2 patients. In our experience the UAT allows the detection of the etiological agent of pneumonia, and also when sputum and/or blood cultures are negative for S. pneumoniae, when the clinical picture is suggestive of alveolar pneumonia.

not available

Acanthamoeba keratitis (AK) is an ocular disease caused by members of a genus of free-living amoebae and it is associated predominantly with contact lens (CL) use. This study reports 55 cases of AK diagnosed in Italy. Genotype identification was carried out by PCR assay followed by sequence analysis of the 18S rRNA gene using the genus specific primers JDP1 and JDP2. Genotype assignment was based on phenetic analysis of the ASA.S1 subset of the small-subunit rRNA gene sequences. The material has been collected at the Polyclinic Tor Vergata of Rome for a total of 19 isolates and at the Polyclinic Hospital of Bari (36 isolates). Thirty-three out of the 55 genetically characterized isolates were assigned to the genotype T4. Ten isolates were identified as belonging to the genotype T15 thus confirming the first association between the genotype T15 and human amoebic keratitis previously described from the same area. We underline the occurrence of the genotype T3 and T11 identified for the first time in the country

The aim of the work was to compare H. pylori clarithromycin-resistance according two methods. Etest was performed on H. pylori isolated from gastric biopsy samples. TaqMan Real-Time-PCR (RT-PCR) was performed on paraffin-embedded gastric biopsy samples of the same patients. Forty-seven out of 88 strains were resistant to clarithromycin by Etest, whereas RT-PCR detected this resistance on paraffin-embedded specimens of 50 patients. RT-PCR performed on paraffin-embedded biopsy specimens of 47 patients infected with H. pylori resistant to clarithromycin as detected by Etest, revealed the presence of a resistant strain only in 40 samples. RT-PCR performed on samples of 41 patients harbouring clarithromycin-susceptible H. pylori strains showed the presence of 31 susceptible and 10 resistant strains. RT-PCR detected 18 cases with heteroresistant status. The difference between the two tests in detecting clarithromycin-resistance was not statistically significant even if RT-PCR detected more resistant cases. The genotyping resistance on paraffin-embedded gastric biopsy specimens may be used to establish resistance to clarithromycin before the treatment when culture and susceptibility testing are not available. In case of failure of an empirical clarithromycin-based triple antimicrobial treatment, RT-PCR performed on paraffin-embedded biopsy sample will establish the primary resistance to clarithromycin. In addition, this test can be useful for epidemiological investigation as well as for monitoring the evolution of clarithromycin resistance along the time. The recommended first-line treatment for H. pylori infection is a proton pump inhibitor combined with amoxicillin and clarithromycin. Unfortunately, primary clarithromycin resistance is increasing worldwide, and it is regarded as the main factor reducing the efficacy of eradication therapy [1-3]. In vitro clarithromycin susceptibility testing are currently based on the agar dilution method or gradient diffusion susceptibility testing (Etest) performed on H. pylori isolated from biopsy specimens [4]. Even if agar dilution is the method recommended by Clinical Laboratory Standard Institute (CLSI), this method and Etest have yielded similar results [5]. Furthermore, the sensitivity of the culture of H. pylori from gastric biopsy samples may be reduced, even in expert hands, by strain fastidiousness, by low bacterial density due to past antibiotic treatments, by loss of viability or overgrowth of contaminating bacteria deriving from delayed transport of the specimens [4]. Recently, a polymerase chain reaction has been developed as an alternative tool for the detection of clarithromycin resistance not only on gastric biopsy specimens but also on paraffin-embedded gastric biopsy samples, thus offering the possibility of performing studies on archival materials [6-8]. This technique accurately assesses mutations in the peptidyltransferase region encoded in domain V of H. pylori 23S ribosomal RNA gene conferring clarithromycin resistance. Despite a dozen of point mutations having been identified, three of them – namely A2143G, A2142G, A2142C – have been shown to be responsible for 90% of cases of primary clarithromycin resistance in Western countries. Moreover, PCR is more sensitive in detecting mixed populations with clarithromycin susceptible and clarithromycin resistant strains (i.e. heteroresistant status) in the gastric biopsy samples [7-9]. The aim of this study was to compare clarithromycin resistance evaluated by Etest performed on H. pylori isolated from gastric biopsy samples with the results obtained by TaqMan Real-Time-PCR (RT-PCR) performed on paraffin-embedded gastric biopsy samples of the same patients.

Objective Resistance to antibiotics is the major cause of treatment failure of Helicobacter pylori infection. A study was conducted to assess prospectively the antibacterial resistance rates of H pylori in Europe and to study the link between outpatient antibiotic use and resistance levels in different countries. Design Primary antibiotic resistance rates of H pylori were determined from April 2008 to June 2009 in 18 European countries. Data on yearly and cumulative use over several years of systemic antibacterial agents in ambulatory care for the period 2001e8 were expressed in Defined Daily Doses (DDD) per 1000 inhabitants per day. The fit of models and the degree of ecological association between antibiotic use and resistance data were assessed using generalised linear mixed models. Results Of 2204 patients included, H pylori resistance rates for adults were 17.5% for clarithromycin, 14.1% for levofloxacin and 34.9% for metronidazole, and were significantly higher for clarithromycin and levofloxacin in Western/Central and Southern Europe (>20%) than in Northern European countries (<10%). Model fit improved for each additional year of antibiotic use accumulated, but the best fit was obtained for 2005. A significant association was found between outpatient quinolone use and the proportion of levofloxacin resistance (p¼0.0013) and between the use of longacting macrolides only and clarithromycin resistance (p¼0.036). Conclusion In many countries the high rate of clarithromycin resistance no longer allows its empirical use in standard anti-H pylori regimens. The knowledge of outpatient antibiotic consumption may provide a simple tool to predict the susceptibility of H pylori to quinolones and to macrolides and to adapt the treatment strategies

Background. Gram-negative bacteria susceptible only to colistin (COS) are emerging causes of severe nosocomial infections, reviving interest in the use of colistin. However, consensus on the most effective way to administer colistin has not yet been reached. Methods. All patients who had sepsis due to COS gram-negative bacteria or minimally susceptible gram-negative bacteria and received intravenous colistimethate sodium (CMS) were prospectively enrolled. The CMS dosing schedule was based on a loading dose of 9 MU and a 9-MU twice-daily fractioned maintenance dose, titrated on renal function. For each CMS course, clinical cure, bacteriological clearance, daily serum creatinine clearance, and estimated creatinine clearance were recorded. Results. Twenty-eight infectious episodes due to Acinetobacter baumannii (46.4%), Klebsiella pneumoniae (46.4%), and Pseudomonas aeruginosa (7.2%) were analyzed. The main types of infection were bloodstream infection (64.3%) and ventilator-associated pneumonia (35.7%). Clinical cure was observed in 23 cases (82.1%). Acute kidney injury developed during 5 treatment courses (17.8%), did not require renal replacement therapy, and subsided within 10 days from CMS discontinuation. No correlation was found between variation in serum creatinine level (from baseline to peak) and daily and cumulative doses of CMS, and between variation in serum creatinine level (from baseline to peak) and duration of CMS treatment. Conclusions. Our study shows that in severe infections due to COS gram-negative bacteria, the high-dose, extended-interval CMS regimen has a high efficacy, without significant renal toxicity.

Since human papillomavirus (HPV) is the central casual factor in cervical cancer, understanding the epidemiology and geographical area distribution of the most prevalent HPV genotypes constitutes an important step towards development of strategies of prevention.

Free-living amoebae (FLA) are protozoa ubiquitous in Nature, isolated from a variety of environments worldwide. In addition to their natural distribution, some species have been found to be pathogenic to humans. In the present study a survey was conducted in order to evaluate the presence and to characterize at molecular level the isolates of amoebic organisms collected from different water sources in Italy. A total of 160 water samples were analyzed by culture and microscopic examination. FLA were found in 46 (28.7%) of the investigated water samples. Groundwater, well waters, and ornamental fountain waters were the sources with higher prevalence rates (85.7%, 50.0%, and 45.9%, respectively). Identification of FLA species/genotypes, based on the 18S rDNA regions, allowed to identify 18 (39.1%) Acanthamoeba isolates (genotypes T4 and T15) and 21 (45.6%) Vermamoeba vermiformis isolates. Other FLA species, including Vahlkampfia sp. and Naegleria spp., previously reported in Italy, were not recovered. The occurrence of potentially pathogenic free-living amoebae in habitats related to human population, as reported in the present study, supports the relevance of FLA as a potential health threat to humans.

PURPOSE: Collection of an endometrial specimen for investigating infectious agents in the endometrial cavity is an invasive technique that is at times difficult and painful. In order to avoid the need for endometrial sampling in the cases of suspected or evident endometrial pathology, the aim of this study is to investigate the reliability of cervical cultures for detecting infectious agents present at the endometrial level, comparing the results between cervical cultures and endometrial cultures in women with clinical signs of endometrial inflammation. METHODS: In a prospective diagnostic study, in the period from January 2009 to October 2010, we enrolled 404 women referred to the Department of Obstetrics and Gynecology for diagnostic hysteroscopy. All the patients underwent cervical and endometrial sampling. Cultures for common bacteria, Neisseria gonorrhoeae, yeast, and Ureaplasma urealyticum were performed. RESULTS: The most frequent infectious agents detected at the endometrial level were common bacteria, which accounted for 69% of all cases. In particular, streptococci were found in 27% of cases, and bacteria from intestinal flora (Enterococcus faecalis and Escherichia coli) was recovered in 31% of cases. U. urealyticum was detected in 10% and Mycoplasma in only one patient (0.2% of cases). No cases of N. gonorrhoeae were found. CONCLUSIONS: Cervical culture has a low concordance with endometrial culture. In fact in only 33% of cases was the microorganism found in the cervix the same as that found in the endometrium. These results infer that an endometrial culture is a useful investigative tool for determining the microorganisms in endometrial pathology.

We investigated the presence of Chlamydophila pneumoniae antibodies in 125 patients with cardiovascular disease and in 128 controls. C. pneumoniae antibodies were measured by microimmunofluorescence assay. A significantly high prevalence of IgG C. pneumoniae antibodies at titre &gt; or = 8 was found in patients (84%) in comparison to controls (47.6%). Considering as cut-off the IgG titre &gt; or = 32, 52% of patients with coronaropathies and 18.75% of controls resulted positive (p &lt; 0.0001). IgA C. pneumoniae antibodies were found in patients and controls without statistically significant differences. High C. pneumoniae antibodies (titre &gt; or = 256) were found in 11% of patients with acute myocardial infarction (AMI) and in none of the controls. In patients, the percentage of IgG and IgA seropositivity increased with age and decreased in patients aged &gt; 70 years. Only patients with AMI are at risk of having antibodies against C. pneumoniae (OR = 6.69). None of the known risk factors for cardiovascular disease was significantly associated with C. pneumoniae seropositivity IgG. This is the first report in our area on the possible association of C. pneumoniae IgG seropositivity and acute ischemic events.

Abstract Background: H. pylori antibiotic resistance is an important factor in the treatment failure, therefore is important to know the local pattern of this resistance. Material and Methods: A total of 111 patients were studied. Ninety- one H. pylori strains isolated from patients, including 12 from children, having previous repeated treatment failure and 20 strains were isolated from naïve patients, were studied. Antibiotic susceptibility including those to tigecycline, was determinated by E-Test. Results: In treated adult and children patients the resistance rates were respectively 81% and 91.6% for clarithromycin; 27.8% and 41% for amoxicillin; 67.1% and 16.7% for metronidazole; 38% and 8.3% for levofloxacin; 5.1% and 0% for tetracycline. Primary resistance, in naïve adult patients was 50% for clarithromycin, 10% for amoxicillin, 20% for metronidazole, 30% for levofloxacin and 0% for tetracycline. Tigecycline has shown good activity, in vitro, against H. pylori (MIC90 = 0.064 mg/L). Conclusion: The resistance rates found in H. pylori, in our area, are very high both in naïve and treated patients. Few papers have reported the tigecycline susceptibility in H. pylori. The good activity and the lack of resistance to tigecycline found in our study, may consider this antibiotic a “rescue” therapy, saving the use of other antibiotics such as rifabutin, a drug used for the treatment of tuberculosis.