Hepatitis C Epidemiology: Insights from a Comprehensive Cohort Study in ASST Melegnano and Martesana, Lombardia Region, Northern Italy.

Hepatitis C virus (HCV) infection is a significant public health problem affecting 58 million people worldwide, including 3.9 million in Europe. Many of these infections go undiagnosed because chronic infection is often asymptomatic. This observational cohort study presents a detailed examination of hepatitis C virus epidemiology in Lombardia (Italy) and was conducted within the ASST "Melegnano e della Martesana". The study involved comprehensive HCV screening of 3290 patients accessing the collection points and/or hospitalized in the facilities of the ASST from 20 May 2022 to 13 April 2023. Screening was conducted using serological assays. The prevalence of anti-HCV-positive patients (HCV-Ab) and then HCV-positive patients (RNA) was calculated. Chi-square tests examined the associations between continuous and categorical variables. Logistic regression was used to evaluate the influence of demographic and geographic variables as predictors of HCV positivity. The study revealed an overall HCV-Ab prevalence of 0.912% (CI (0.59-1.24%)) in the examined cohort, of whom 15.15% (two females and three males) were positive for HCV RNA. The prevalence of HCV RNA positivity was 0.152% (CI (0.05-0.35%)). Sex disparity was evident, with male patients exhibiting a higher prevalence compared to females, confirmed by logistic regression (0.0147 vs. 0.0061-OR = 2.44; CI (0.0059-0.0124)). Age stratification indicated an ascending trend in prevalence with age, peaking at 1.35% in individuals aged over 50. These findings underscore the critical need for targeted HCV screening, contributing valuable insights to the global epidemiology of HCV in the era of DAAs.

First report of Kocuria marina bloodstream infection unrelated to a central venous catheter: a mini-review on an emerging and under-recognized opportunistic pathogen.

We report a case of sepsis by Kocuria marina in a 45-year-old woman carrying a midline venous catheter to provide total parenteral nutrition. Despite the finding of K. marina bacteraemia, no bacteria grew from the culture of the catheter tip. As the patient was affected with severe leg ulcers, K. marina bloodstream infection from a skin breakage other than CVC was supposed. K. marina is an emerging opportunistic agent deserving attention and probably under-recognized, as it can be misdiagnosed as Staphylococcus if only conventional microbiological analyses are performed for bacterial identification.

Isolation of Enterobacter aerogenes carrying blaTEM-1 and blaKPC-3 genes recovered from a hospital Intensive Care Unit.

Enterobacter aerogenes has recently emerged as an important hospital pathogen. In this study, we showed the emergence of E. aerogenes isolates carrying the blaKPC gene in patients colonized by carbapenem-resistant Klebsiella pneumoniae strains. Two multiresistant E. aerogenes isolates were recovered from bronchial aspirates of two patients hospitalized in the Intensive Care Unit at the "Santa Maria della Scaletta" Hospital, Imola. The antimicrobial susceptibility test showed the high resistance to carbapenems and double-disk synergy test confirmed the phenotype of KPC and AmpC production. Other investigation revealed that ESBL and blaKPC genes were carried on the conjugative pKpQIL plasmid. This is a relevant report in Italy that describes a nosocomial infection due to the production of KPC beta-lactamases by an E. aerogenes isolate in patients previously colonized by K. pneumoniae carbapenem-resistant. In conclusion, it's necessary a continuous monitoring of multidrug-resistant strains for the detection of any KPC-producing bacteria that could expand the circulation of carbapenem-resistant pathogens.

Urinary Schistosomiasis in an Adolescent Refugee from Africa: An Uncommon Cause of Hematuria and an Emerging Infectious Disease in Europe.

We report a case of urinary schistosomiasis in an adolescent refugee from Gambia (arrived to Italy illegally), who was brought to the Emergency Department of our hospital. The patient complained of gross hematuria and, in the absence of clinical evidence of bacterial urinary infection, was admitted to the pediatric ward, considering his provenience and social setting. An appropriate collection and microscopic analysis of urine samples led to the detection of bilharzia. Much attention should be paid to this emerging disease in Europe by physicians in order to recognize and treat it timely, which could prevent future and higher costs for public health systems and could reduce the potential risk of environmental spreading. In fact, there are some areas in Italy where the parasite can find its intermediate host to complete its lifecycle.

A rare case of infant sepsis due to the emm-89 genotype of Group A Streptococcus within a community-acquired cluster.

Invasive Group A Streptococcus disease is a severe and sometimes life-threatening infection with only few cases reported in literature. We describe the case of a 49-day-old male infant with invasive Group A Streptococcus infection characterized by acute otitis media and development of septicemia within a probably community-acquired cluster. The causative agent resulted to be a rare emm-89 genotype of Streptococcus pyogenes. Group A Streptococcus must be considered responsible for sepsis in newborns and young infants.

Anti-chlamydial IgG Neutralizing Ability in Nonzoonotic Atypical Community Acquired Respiratory Tract Infections.

Chlamydophila pneumoniae is a pathogenic agent, involved in various types of infection. This study has evaluated the ability of IgG antibodies in outpatient, with acute respiratory tract infections from C. pneumoniae, to neutralize in vitro purified elementary bodies of this bacterium, revealing a good neutralizing performance of IgG antibodies.

Selectivity evaluation of a new chromogenic medium to detect group B Streptococcus.

BACKGROUND: Group B Streptococcus (GBS) is an important pathogen that causes serious infections in newborns. Pregnant screening and intrapartum antibiotic prophylaxis are actually the strategies to prevent GBS disease in neonates because vaccination is under investigation. MATERIALS AND METHODS: Simultaneously, 156 isolates of GBS and 156 isolates other than GBS covering 17 different species, were tested to evaluate the selectivity of a new chromogenic medium to screen GBS. RESULTS: The new new chromogenic medium showed an excellent performance, exhibiting a very high level of inclusivity (100%) and exclusivity (96.1%).

Typing of Ochrobactrum anthropi clinical isolates using automated repetitive extragenic palindromic-polymerase chain reaction DNA fingerprinting and matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry.

BACKGROUND: Ochrobactrum anthropi (O. anthropi), is a non-fermenting gram-negative bacillus usually found in the environment. Nevertheless, during the past decade it has been identified as pathogenic to immunocompromised patients. In this study, we assessed the usefulness of the automated repetitive extragenic palindromic-polymerase chain reaction (rep-PCR-based DiversiLab™ system, bioMèrieux, France) and of matrix-assisted laser desorption/ionization-time-of-flight (MALDI-TOF MS) for typing of twentythree O. anthropi clinical isolates that we found over a four-months period (from April 2011 to August 2011) in bacteriemic patients admitted in the same operative unit of our hospital. Pulsed-field gel electrophoresis (PFGE), commonly accepted as the gold standard technique for typing, was also used. Analysis was carried out using the Pearson correlation coefficient to determine the distance matrice and the unweighted pair group method with arithmetic mean (UPGMA) to generate dendogram. RESULTS: Rep-PCR analysis identified four different patterns: three that clustered together with 97% or more pattern similarity, and one whose members showed < 95% pattern similarity. Interestingly, strains isolated later (from 11/06/2011 to 24/08/2011) displayed a pattern with 99% similarity. MALDI-TOF MS evaluation clustered the twentythree strains of O. anthropi into a single group containing four distinct subgroups, each comprising the majority of strains clustering below 5 distance levels, indicating a high similarity between the isolates. CONCLUSIONS: Our results indicate that these isolates are clonally-related and the methods used afforded a valuable contribution to the epidemiology, prevention and control of the infections caused by this pathogen.

Clonal dissemination of Klebsiella pneumoniae ST512 carrying blaKPC-3 in a hospital in southern Italy.

Strains of Klebsiella pneumoniae producing KPC-carbapenemase have emerged as one of the most important multidrug-resistant Gram-negative nosocomial pathogens. Here, we report the first isolation and subsequent dissemination of a K. pneumoniae ST512 producing KPC-3 carbapenemase in a hospital in southern Italy. Isolates were obtained from blood, throat swabs, sputum, catheters, and urine of patients admitted to different hospital wards. Antimicrobial MICs were determined for all isolates by automated systems and confirmed by Etest. Carbapenemase production was confirmed by the modified Hodge test and by a disc synergy test, and carbapenemase genes were investigated by PCR. All isolates were characterized by pulse-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) analysis. Most isolates were multidrug resistant with exception of some isolates intermediately susceptible to gentamicin, tigecycline, and trimethoprim-sulfamethoxazole. PCR analysis showed that isolates harbored the bla(KPC-3) gene associated with bla(TEM) and bla(SVH). PFGE and MLST showed that all isolates belonged to the same ST512 clone recently described in Israel.

In vitro antimicrobial profile of Ureaplasma urealyticum from genital tract of childbearing-aged women in Northern and Southern Italy.

Ureaplasma urealyticum is an opportunistic pathogen during pregnancy and in newborns. Other clinical problems related to U. urealyticum infections are: no susceptibility to cell wall-active drugs, limits of antibiotic treatment in pregnancy, and spread of antimicrobial resistance. In addition, the results of antimicrobial susceptibility against U. urealyticum from various countries are few and controversial. The antimicrobial susceptibility of U. urealyticum, isolated from cervical swabs and collected from outpatient childbearing-aged women in Italy from 2009 to 2012, was performed against fluoroquinolones, macrolides, streptogramin and tetracyclines, using an available biochemical commercial kit and a specific solid culture medium, to improve the therapeutic management of these pathogenic agents. Ureaplasma urealyticum was detected in 49.4% of samples, but significant bacterial load was revealed in 29.8%. In vitro tetracyclines showed the best activity against U. urealyticum, followed by streptogramin, macrolides, and fluoroquinolones.

Rapid and reliable MALDI-TOF mass spectrometry identification of Candida non-albicans isolates from bloodstream infections.

Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) fingerprinting has recently become an effective instrument for rapid microbiological diagnostics and in particular for identification of micro-organisms directly in a positive blood culture. The aim of the study was to evaluate a collection of 82 stored yeast isolates from bloodstream infection, by MALDI-TOF MS; 21 isolates were identified also directly from positive blood cultures and in the presence of other co-infecting micro-organisms. Of the 82 isolates grown on plates, 64 (76%) were correctly identified by the Vitek II system and 82 (100%) by MALDI-TOF MS; when the two methods gave different results, the isolate was identified by PCR. MALDI-TOF MS was unreliable in identifying two isolates (Candida glabrata and Candida parapsilosis) directly from blood culture; however, direct analysis from positive blood culture samples was fast and effective for the identification of yeast, which is of great importance for early and adequate treatment.

Molecular and phenotypic characterization of methicillin-resistant Staphylococcus aureus from surgical site infections.

BACKGROUND: Nosocomial infections represent an important problem for the health of hospitalized patients. Peri-operative infections--those occurring during surgery or in the post-operative period--account for 15%-20% of cases. Most surgical site infections (SSIs) are caused by endogenous gram-positive microorganisms, in particular, Staphylococcus aureus, S. epidermidis, and other coagulase-negative staphylococci that are part of the flora of the skin. METHODS: A retrospective study was conducted from January 2006 to December 2010 to describe the epidemiology of methicillin-resistant S. aureus (MRSA) in SSIs. The MRSA isolates were analyzed by a combination of two genotyping methods: SCCmec and pulsed-field gel electrophoresis (PFGE). Also, biofilm-forming ability was analyzed for all isolates as an indicator of their ability to persist despite antibiotic treatment. RESULTS: During the study period, 1,793 swabs from SSIs were analyzed, and S. aureus was identified in 318/987 positive specimens (32%). Methicillin resistance was observed in 10% of the S. aureus isolates (n=33). Analysis by PFGE revealed that isolates with the same SCCmec type were unrelated. Instead, biofilm-forming ability tests showed that SCCmec type I MRSA had the highest ability to form a film. CONCLUSIONS: The strains analyzed in our study showed a homogeneous pattern of SCCmec type. The difference in ability to produce biofilm between strains of SCCmec type I and isolates with other SCCmecs was substantial. This virulence factor could have critical implications for the formation and persistence of chronic SSIs.

Different mutations in mucA gene of Pseudomonas aeruginosa mucoid strains in cystic fibrosis patients and their effect on algU gene expression.

Alginate biosynthesis in Pseudomonas aeruginosa is a highly regulated process in which algU and mucA genes are key elements. Mutations in mucA gene determine alginate operon overexpression and exopolysaccharide overproduction. In our study, 119 strains of P. aeruginosa were isolated from sputa of 96 cystic fibrosis patients and 84/119 showed nonmucoid phenotype, while 35/119 showed mucoid phenotypes. mucA gene was amplified and sequenced in all strains revealing mutations in 29/35 mucoid strains (82%) and in one non-mucoid strain. 4/29 strains showed mutations never described that generated premature stop and much shorter MucA proteins. In all mutated strains, algU gene expression was analyzed to determine if mutations in mucA, resulting in a strong loss of its protein, could significantly influence its function and subsequently the biosynthetic pathways under algU control. Analysis of algU expression disclosed that the length significantly affects the expression of genes involved in the production of alginate and in the motility and hence survival of P. aeruginosa strains in cystic fibrosis lungs.

Ambroxol influences voriconazole resistance of Candida parapsilosis biofilm.

The ability to form biofilm on different surfaces is typical of most Candida species. Microscopic structure and genetic aspects of fungal biofilms have been the object of many studies because of very high resistance to antimycotic agents because of the scarce permeability of the external matrix and to the alterations in cell metabolism. In our study, 31 isolates of Candida parapsilosis, isolated from bloodstream infections, were tested for their ability to produce biofilm and were found to be good producers. The susceptibility to voriconazole, assayed by colorimetrical XTT assay, revealed a very elevated minimum inhibitory concentrations for sessile cells in comparison with planktonic ones. The addition of ambroxol, a mucolytic agent, increased the susceptibility of biofilm forming cells to voriconazole. Expression of the efflux pump genes CDR and MDR was analyzed in biofilms alone or treated with ambroxol, evidencing a role of ambroxol in the expression of genes involved in azole resistance mechanisms of C. parapsilosis biofilms. In conclusion, our data seem to encourage the use of different substances in combination with classical antimycotics, with the aim of finding a solution to the increasing problem of the resistance of biofilms formed on medical devices by nonalbicans Candida species.

On the possibility that H1 histone interaction with DNA occurs through phosphates connecting lysine and arginine side chain groups.

Gel filtration and velocity of sedimentation analyses on native and on lysine- and arginine-modified forms of the annelid worm Chaetopterus variopedatus sperm H1 histone indicate that anion-mediated lysine-arginine interactions play a relevant role in the stabilization of the oligomeric states of the molecule. CD spectroscopy shows that phosphate anions are at least an order of magnitude more efficient than chloride as negatively charged groups connecting H1 lysines and arginines. Acetylation of lysines, although not altering grossly the H1 properties, causes a tenfold decrease of the structuring efficiency of phosphates. This suggests that DNA phosphates may be sandwiched between lysine and arginine groups of H1 histone when this molecule binds to chromatin, constituting a relevant parameter for the reciprocal stabilization of the protein and of the chromatin higher order structures.

Methylation profile of P. lividus sea urchin genes during development.

Methylation pattern has been studied in two genes of sea urchin Paracentrotus lividus using sodium bisulfite method to understand the possible role of DNA methylation during invertebrate development. Three regions of the gene for the hatching enzyme have been analyzed and all of them resulted unmethylated in embryos at different stages of development. Four CpG rich regions have been studied in the gene for DNA methyltransferase: upstream, upstream-exon1, intron 1 and exon 20. The upstream-exon 1 region is always unmethylated, while intron 1 and exon 20 are heavy methylated. Only the upstream fragment changed its pattern of methylation during development. For none of the studied regions the reported data show a general direct correlation between gene expression and methylation process during development.