The occurrence of Malassezia spp. bloodstream infections (BSIs) in neonatal intensive care unit was evaluated by using pediatric Isolator, BacT/Alert systems and central venous catheter (CVC) culture. The efficacy of BacT/Alert system in detecting Malassezia was assessed by conventional procedures, culturing 1 ml of bottle content before incubation and by studying the survival of Malassezia spp. strains in BacT/Alert bottles. Of the 492 neonates enrolled, blood was collected by pediatric Isolator (290 patients; group I) or by BacT/Alert bottles (202 patients; group II). The survival of Malassezia furfur and Malassezia pachydermatis in BacT/Alert bottles was evaluated by culturing the inoculum suspension (from 106 to 10 colony-forming units, cfu/ml) and assessing the cfu/ml for 15 days. In total, 15 Malassezia BSIs were detected, of which six (2.1%) from both blood and CVC culture in Dixon agar (DixA) in patients belong to group I (blood collected by paediatric Isolator tube) and nine (4.4%) only from CVC culture in DixA in patients of group II (blood collected by BacT/Alert bottle). Only one patient (0.5%) from group II scored positive for M. furfur also by culturing in DixA 1 ml blood content of BacT/Alert bottle before incubation in BacT/Alert system.M. furfur population size in BacT/Alert bottles decreased during the incubation time, whereas that of M. pachydermatis increased. The BacT/Alert system detected M. pachydermatis even at very low concentration (i.e., 10 cfu/ml) but not any positive blood culture for M. furfur. For a correct diagnosis of Malassezia furfur BSI, the blood should be culture in lipid-enriched fungal medium, and the BacT/Alert system implemented by adding lipid substrates to increase the method sensibility. Finally, CVC cultures on lipid-supplemented media may be proposed as a routine procedure to diagnose the Malassezia fungemia.

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.

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.

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.

Human papillomavirus (HPV) is considered the most important risk factor for the development of ano-genital region cancer in both women and men. Whereas low-risk genotypes are responsible for cutaneous and genital lesions, high-risk genotypes are associated with ano-genital cancer. The aim of this study was to retrospectively analyse the prevalence, genotype distribution and temporal dynamics of HPV infection in 2312 specimens from 2312 subjects (2149 women and 163 men) who attended the laboratory of molecular biology, UOC Microbiology and Virology, Azienda Ospedaliera-Universitaria, Policlinico of Bari, Italy. HPV DNA detection and genotyping was performed using a multiplex real-time PCR assay. In all, 1123/2312 subjects (48.57%) resulted positive for HPV DNA. In particular, HPV DNA was detected in (1056) 49.14% of females and (67) 41.10% of males. HPV co-infections were detected in 565 (24.44%) patients. High-risk and low-risk HPV genotypes were detected in 887 (38.37%) and 600 (25.95%) patients, respectively. The most prevalent HPV genotypes were HPV-42 (10.29%), HPV-16 (8.56%), HPV-31 (7.40%) and HPV-53 (7.14%). Statistically significant differences between female and male patients were not detected. Moreover, HPV prevalence remained constant in time while HPV-16, but not HPV-6, 11 and 18, showed a decreasing trend from 2013 (11.24%) to 2016 (6.67%). Other HPV genotypes showed some complex and different patterns. Our data showed an unusually high frequency of HPV-42 and a high prevalence of HPV infection in the patients analysed. Although evidence of a decreasing trend of HPV-16 could be a consequence of anti-HPV vaccination, corroboration from further studies will be needed. Moreover, the small number of studied males and the similarity to females in terms of HPV prevalence suggest that more active HPV screening and anti-HPV vaccination in the male population should be considered important tools to eliminate HPV sexual transmission.

Background: The aim of this study was the rapid identification of blaKPC gene in 38 Klebsiella pneumoniae clinical isolates with reduced susceptibility to carbapenems. The modified Hodge Test (MHT) was carried out to phenotypically determine whether resistance to carbapenems was mediated by a carbapenemase. The detection of the blaKPC gene was performed by real-time acid nucleic sequence-based amplification (NASBATM), specifically designed for the detection of KPC RNA target. Results: Thirty-two/38 isolates evaluated by MHT showed the production of carbapenemases, while all the strains exhibited the production of KPC by inhibition test with phenylboronic acid (the combined disk test with IPM/IPM plus phenylboronic acid). The detection of blaKPC gene by Nuclisens EasyQ KPC yielded positive results in 38/38 (100%) strains. The presence of blaKPC gene was confirmed in all K. pneumoniae isolates when tested by the gold standard PCR assay. Conclusions: In consideration of the serious challenge represented by infections due to K. pneumoniae it appears necessary the rapid identification of carbapenemases in clinical settings as it is made possible by the use of NASBATM assay.

PURPOSE: Genital tract infections are globally a major cause of morbidity in sexually active individuals. The aim of this study was to investigate the prevalence and associations of co-infections of Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma hominis (MH), Mycoplasma genitalium, Ureaplasma urealyticum (UU) and Ureaplasma parvum (UP) in specimens collected from female (SF) and male (SM) patients. METHODS: 1575 samples from 1575 individuals from the geographical area around Bari, Apulia region in Southern Italy, were collected and analyzed by a multiplex Real-Time PCR (mRT-PCR) (AnyplexTM II STI-7, Seegene, Inc., Seoul, Korea) assay. RESULTS: 455/1575 (28.89%) samples resulted positive for at least one of the targets named above. Statistically significant differences in prevalence of the pathogens between SF and SM were not detected except for UP (24.92% in SF vs 8.91% in SM). Prevalence of co-infections was 6.84 and 3.96% in SF and SM, respectively. Moreover, MH presence in SF, but not in SM, was associated with UU and UP. CONCLUSIONS: Our data suggest different patterns of infections between females and male and the importance of an increased vigilance of sexually transmitted pathogens to reduce the burden on general population and the sequelae or the complications on reproductive organs.

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.

The tuberculosis of the ear is rare, and in most cases the clinical picture resembles that of a chronic otitis media. The diagnosis is often delayed, and this can lead to irreversible complications such as hearing loss and/or facial paralysis. In view of its rare occurrence, we report a case of primary tuberculous otitis media in a 87-year-old female patient. The diagnosis was made on the basis of both histological and microbiological findings. In particular, gene amplification techniques such as real-time polymerase chain reaction are useful method for rapid diagnosis and detecting tuberculous bacilli usually present at very low number. Early diagnosis is essential for the prompt institution of antituberculous therapy.