A urokinase-associated outbreak of Ralstonia mannitolilytica bloodstream infections in haemodialysis patients in north-eastern Italy, January to April 2023.

An outbreak of Ralstonia mannitolilytica bloodstream infections occurred in four hospitals in north-eastern Italy, involving 20 haemodialysis patients with tunnelled central vascular catheter access. We identified as the outbreak source a batch of urokinase vials imported from India contaminated with R. mannitolilytica. Whole genome sequences of the clinical and urokinase strains were highly related, and only urokinase-treated patients were reported with R. mannitolilytica infections (attack rate = 34%; 95% confidence interval: 22.1-47.4). Discontinuation of the contaminated urokinase terminated the outbreak.

A combined role for low vitamin D and low albumin circulating levels as strong predictors of worse outcome in COVID-19 patients.

PURPOSE: We aimed to assess the combined role of vitamin D and albumin serum levels as predictors of COVID-19 disease progression. METHODS: We conducted a prospective observational study on adult patients hospitalized for SARS-CoV-2 pneumonia (March-September 2020). Vitamin D and albumin serum levels were measured on admission. These variables were categorized in albumin < 3.5 or ≥ 3.5 g/dL and vitamin D < 30 ng/mL or ≥ 30 ng/mL. We excluded patients with known bone diseases, renal failure, hypercalcemia and/or treated with antiepileptic drugs and steroids, and patients who received previous vitamin D supplementation. A composite outcome including any ventilatory support, PaO2/FiO2 ratio, and 60-day mortality was defined. RESULTS: Sixty-nine patients were enrolled, of whom 50% received non-invasive (NIV) or invasive mechanical ventilation (IMV), 10% died, whereas 89% and 66% presented low albumin and low vitamin D serum levels, respectively. No correlation between vitamin D and albumin levels was found. In multivariable logistic regression analyses adjusted for sex and age-corrected comorbidities, patients having albumin < 3.5 g/dL and vitamin D < 30 ng/mL showed a significant increased risk for all study outcomes, namely NIV/IMV (OR 3.815; 95% CI 1.122-12.966; p = 0.032), NIV/IMV or death (OR 3.173; 95% CI 1.002-10.043; p = 0.049) and PaO2/FIO2 ≤ 100 (OR 3.410; 95% CI 1.138-10.219; p = 0.029). CONCLUSION: The measurement of both vitamin D and serum albumin levels on COVID-19 patients' admission, and their combined evaluation, provides a simple prognostic tool that could be employed to guide prompt clinical decisions.

Discriminatory Value of Adiponectin to Leptin Ratio for COVID-19 Pneumonia.

Purpose: Obesity is a risk factor for severe coronavirus disease 2019 (COVID-19). Circulating adipokines have been associated with inflammatory burden and amplified or dysregulated immune responses. This study aimed to evaluate the discriminatory ability of adipokines to identify COVID-19 pneumonia and to assess disease severity. Methods: We conducted an observational case-control study, with a prospective design, and recruited patients with diagnosis of COVID-19 pneumonia (n = 48) and healthy controls (n = 36), who were matched by age, sex, and BMI. Leptin, adiponectin, IL-6, and TNF-α were measured by ELISA. Results: Patients with COVID-19 pneumonia had higher levels of leptin, lower adiponectin/leptin (Adpn/Lep) ratio, and higher expression of IL-6. Leptin had an acceptable discriminatory accuracy for COVID-19 pneumonia in patients with BMI >30 (AUC 0.74 [0.58, 0.90]) with a cutoff of 7852 pg/mL and it was associated with maximum respiratory support. By contrast, Adpn/Lep had an excellent discriminatory accuracy for COVID-19 pneumonia in patients with BMI <25 (AUC 0.9 [0.74, 1.06]) with a cutoff of 2.23. Conclusion: Our data indicate that high Adpn/Lep (>2.23) in lean patients is consistent with a state of good health, which decreases in case of inflammatory states, ranging from adipose tissue dysfunction with low-grade inflammation to COVID-19 pneumonia.

Case report: Long-lasting SARS-CoV-2 infection with post-COVID-19 condition in two patients with chronic lymphocytic leukemia: The emerging therapeutic role of casirivimab/imdevimab.

Post-coronavirus disease 2019 (post-COVID-19) condition, previously referred to as long COVID, includes a post-acute syndrome defined by the presence of non-specific symptoms occurring usually 3 months from the onset of the acute phase and lasting at least 2 months. Patients with chronic lymphocytic leukemia (CLL) represent a high-risk population for COVID-19. Moreover, the response to SARS-CoV-2 vaccination is often absent or inadequate. The introduction of monoclonal antibodies (mAbs) in the treatment landscape of COVID-19 allowed to reduce hospitalization and mortality in mild-moderate SARS-CoV-2 infection, but limited data are available in hematological patients. We here report the effective use of casirivimab/imdevimab (CI) in the treatment of two CLL patients with persistent infection and post-COVID-19 condition. Full genome sequencing of viral RNA from nasopharyngeal swabs was performed at the time of COVID-19 diagnosis and before the administration of CI. Both patients experienced persistent SARS-CoV-2 infection with no seroconversion for 8 and 7 months, respectively, associated with COVID symptoms. In both cases after the infusion of CI, we observed a rapid negativization of the nasal swabs, the resolution of post-COVID-19 condition, and the development of both the IgG against the trimeric spike protein and the receptor-binding domain (RBD) of the spike protein. The analysis of the viral genome in the period elapsed from the time of COVID-19 diagnosis and the administration of mAbs showed the development of new mutations, especially in the S gene. The genome variations observed during the time suggest a role of persistent SARS-CoV-2 infection as a possible source for the development of viral variants. The effects observed in these two patients appeared strongly related to passive immunity conferred by CI treatment permitting SARS-CoV-2 clearance and resolution of post-COVID-19 condition. On these grounds, passive anti-SARS-CoV-2 antibody treatment may represent as a possible therapeutic option in some patients with persistent SARS-CoV-2 infection.

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.

What are school children in Europe being taught about hygiene and antibiotic use?

e-Bug is a pan-European antibiotic and hygiene teaching resource that aims to reinforce awareness in school children of microbes, prudent antibiotic use, hygiene and the transmission of infection. Prior to the production of the resource, it was essential to examine the educational structure across each partner country and assess what school children were being taught on these topics. A questionnaire was devised for distribution to each European partner (Belgium, Czech Republic, Denmark, England, France, Greece, Italy, Poland, Portugal and Spain), exploring their educational structure and examining educational resources or campaigns currently available. From the data collected it was evident that the majority of European schools have structured hand hygiene practices in place from a young age. The curricula in all countries cover the topic of human health and hygiene, but limited information is provided on antibiotics and their prudent use. School educational resources that link to the national curriculum and implement National Advice to the Public campaigns in the classroom are limited. The Microbes en question mobile health education campaign in France is an example of a successful children's education campaign and an innovative programme. Evaluation of the impact of school education on attitude and change of behaviour is also limited throughout many European countries. Not enough is currently being done across Europe to educate school children on the importance of appropriate antibiotic use and antibiotic resistance. The data from this research were used to develop e-Bug, a European Union-funded antibiotic and hygiene teaching resource.

Development of an educational resource on microbes, hygiene and prudent antibiotic use for junior and senior school children.

Health promotion interventions aimed at children and young people have the potential to lay the foundations for healthy lifestyles. One such intervention, e-Bug, aims to provide schoolchildren with knowledge of prudent antibiotic use and how to reduce the spread of infection. Many children and schools approach learning in different ways; therefore, it is essential to research school needs and the variety of learning styles when creating any school resources. This article outlines the process involved during the development of a pan-European educational resource, and identifies the final pack layout, based on feedback from teacher focus groups, student questionnaires and European partner discussions.

Overview of e-Bug: an antibiotic and hygiene educational resource for schools.

Antibiotic resistance is an increasing community problem and is related to antibiotic use. If antibiotic use could be reduced, the tide of increasing resistance could be stemmed. e-Bug is a European project involving 18 European countries, partly funded by The Directorate-General for Health and Consumers (DG SANCO) of the European Commission. It aims to develop and disseminate across Europe a junior and senior school teaching pack and web site (hosting the lesson plans and complementary games) that teach young people about prudent antibiotic use, microbes, transmission of infection, hygiene and vaccines. The aim of e-Bug is to increase young people's understanding, through enjoyable activities, of why it is so important to use antibiotics correctly in order to control antibiotic resistance, and to have good hand and respiratory hygiene to help reduce the spread of infection. Within the senior school pack the sexual transmission of infections has also been included, as the peak age of chlamydial infection is in 16-24 year olds. Teachers, young people and the consortium of 18 countries were closely involved with agreeing learning outcomes and developing the resource activities. Young people helped create the characters and microbe artwork. The resources have been translated, adapted for and disseminated to schools across 10 countries in Europe, and endorsed by the relevant government departments of health and education. The web site has been accessed from >200 countries. The resources will be translated into all European Union languages, and have been used to promote European Antibiotic Awareness Day and better hand and respiratory hygiene during the influenza pandemic in 2009.

Developing e-Bug web games to teach microbiology.

As a complement to the e-Bug teaching pack, two e-Bug games were developed to provide content that aimed to entertain as well as to educate. A set of agreed learning outcomes (LOs) were provided by the scientific partners of the e-Bug Project and the games were developed using user-centred design techniques (the needs, wants and limitations of the potential game players were assessed at each stage of the design process). The e-Bug games were designed for two age groups: Junior (9-12 year olds); and Senior (13-15 year olds). A study using focus groups was done to gain an understanding as to the types of games enjoyed by the target users. According to the preliminary study, the Junior Game was developed as a platform game and the Senior Game was developed as a story-based detective game. The Junior Game consists of five levels, each associated with a set of LOs. Similarly, the Senior Game consists of four missions, each comprising five stages using problem-based learning techniques and LOs. In this paper, the process of development for each game is described in detail and an illustration is provided of how each game level or mission addresses the target LOs. Development of the games used feedback acquired from children in four schools across the UK (Glasgow, London and two in Gloucester). The children were selected according to their willingness to participate. European Partners of the e-Bug Project also provided further support, translation and requests for modifications. The knowledge gained of LOs and further evaluation of the games is continuing, and preliminary results are in press. The final versions of the games, translated into 11 European languages, are available online via www.e-bug.eu.

High fecal calprotectin levels are associated with SARS-CoV-2 intestinal shedding in COVID-19 patients: A proof-of-concept study.

BACKGROUND: One third of coronavirus disease 2019 (COVID-19) patients have gastrointestinal symptoms. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA has been detected in stool samples of approximately 50% of COVID-19 individuals. Fecal calprotectin is a marker of gastrointestinal inflammation in the general population. AIM: To investigate if fecal calprotectin correlates with SARS-CoV-2 intestinal shedding in COVID-19 patients with pneumonia. METHODS: Patients with SARS-CoV-2 pneumonia admitted to the Infectious Disease Unit (University Hospital of Trieste, Italy) from September to November 2020 were consecutively enrolled in the study. Fecal samples were collected and analyzed for quantification of fecal calprotectin (normal value < 50 mg/kg) and SARS-CoV-2 RNA presence by polymerase chain reaction (PCR). Inter-group differences were determined between patients with and without diarrhea and patients with and without detection of fecal SARS-CoV-2. RESULTS: We enrolled 51 adults (40 males) with SARS-CoV-2 pneumonia. Ten patients (20%) presented with diarrhea. Real-time-PCR of SARS-CoV-2 in stools was positive in 39 patients (76%), in all patients with diarrhea (100%) and in more than two thirds (29/41, 71%) of patients without diarrhea. Obesity was one of the most common comorbidities (13 patients, 25%); all obese patients (100%) (P = 0.021) tested positive for fecal SARS-CoV-2. Median fecal calprotectin levels were 60 mg/kg [interquartile range (IQR) 21; 108]; higher fecal calprotectin levels were found in the group with SARS-CoV-2 in stools (74 mg/kg, IQR 29; 132.5) compared to the group without SARS-CoV-2 (39 mg/kg, IQR 14; 71) (P < 0.001). CONCLUSION: High fecal calprotectin levels among COVID-19 patients correlate with SARS-CoV-2 detection in stools supporting the hypothesis that this virus can lead to bowel inflammation and potentially to the 'leaky gut' syndrome.

Performance of Vitek 2 in antimicrobial susceptibility testing of Pseudomonas aeruginosa isolates with different mechanisms of beta-lactam resistance.

A total of 78 isolates of Pseudomonas aeruginosa grouped according to the phenotype for ceftazidime and imipenem susceptibility/resistance were used to assess the accuracy of the Vitek 2 system in antimicrobial susceptibility testing. Comparisons were made with a MIC gradient test for piperacillin-tazobactam, ceftazidime, aztreonam, imipenem, meropenem, gentamicin, and ciprofloxacin. For the total of 546 isolate-antimicrobial combinations tested, the category agreement was 83.6%, with 2.0, 1.6, and 12.8% very major, major, and minor errors, respectively. Vitek 2 accuracy was influenced differently by the mechanism responsible for resistance, and interpretation of the results in relation to phenotype could improve the performance of the system.

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.

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.

Triazole resistance in Aspergillus fumigatus isolates from patients with cystic fibrosis in Italy.

BACKGROUND: Aspergillus fumigatus is frequently recovered from respiratory secretions of cystic fibrosis (CF) patients. Azole resistance has been increasingly reported. OBJECTIVES: To assess the prevalence of azole resistance in A. fumigatus isolates from patients followed by two CF centers of northern Italy. METHODS: 423 isolates (220 patients) were screened for azole resistance. Resistance was confirmed with the EUCAST method and cyp51A gene sequencing. Microsatellite genotyping was performed and results were compared with those of environmental resistant isolates. RESULTS: No resistance was detected in one center, while 8.2% of the patients of the other center harbored resistant isolates. The TR34/L98H alteration in the cyp51A gene, present in seven cases, resulted associated with poor in-vitro activity of all tested azoles. CONCLUSIONS: The environmental origin of the resistance seems to be probable since azole resistance was found also in naïve patients and an identical microsatellite genotype in clinical and environmental isolates was observed.

Cross-reactivity of Nocardia spp. in the fungal (1-3)-ß-d-glucan assay performed on cerebral spinal fluid.

Cerebral spinal fluid from a patient affected by a brain abscess caused by Nocardia abscessus gave a positive result for (1-3)-ß-d-glucan (BG) assay, in absence of any fungal infection. This study aimed to assess whether Nocardia spp. show cross-reactivity with BG assay. All Nocardia spp. analyzed provided positive reactions.

Activities of 16-membered ring macrolides and telithromycin against different genotypes of erythromycin-susceptible and erythromycin-resistant Streptococcus pyogenes and Streptococcus pneumoniae.

OBJECTIVES: To test four 16-membered macrolides (josamycin, spiramycin, midecamycin and rokitamycin) along with other compounds in the same class (erythromycin, clarithromycin, roxithromycin and azithromycin) plus clindamycin and telithromycin, against Streptococcus pyogenes and Streptococcus pneumoniae isolates with well-characterized resistance genotypes. METHODS: Four hundred and eighty-six isolates of S. pyogenes and 375 isolates of S. pneumoniae were assayed for their macrolide susceptibilities and investigated by PCR to detect their different erythromycin resistance genes. All strains had been isolated over the period 2002-2003 from specimens of different human origin obtained in 14 different Italian centres. RESULTS: All 16-membered macrolides showed very low MICs (MIC(50)s and MIC(90)s, < or =0.06-0.5 mg/L) for the erythromycin-susceptible isolates and for those with the M phenotype, but the telithromycin MICs for the M-type isolates were at least four times higher (MIC(90)s, 0.5 mg/L). In S. pyogenes, the MIC(50)s of 16-membered macrolides for the cMLS(B) isolates were > or = 256 mg/L, whereas that for telithromycin was 4 mg/L; the MIC(50)s of 16-membered macrolides and telithromycin ranged from < or = 0.06 to 0.5 mg/L for the iMLS(B) isolates with erm(A) and from 0.12 to > or = 256 mg/L for those with erm(B). In S. pneumoniae, the MIC(50)s of the 16-membered macrolides for the cMLS(B) isolates ranged from 0.5 to 128 mg/L, whereas for the iMLS(B) isolates their values ranged from < or = 0.06 to 4 mg/L; the MIC(50)s and MIC(90)s of telithromycin for both the cMLS(B) and the iMLS(B) isolates ranged from < or = 0.06 to 0.12 mg/L. CONCLUSIONS: MICs ranged for all the drugs, except telithromycin, from < or = 0.06 to > or = 256 mg/L, with 15% to 30% resistant S. pyogenes for all drugs tested except clindamycin (8%) and telithromycin (5.4%) and 10% to 40% resistant S. pneumoniae for all drugs tested except telithromycin (0.3%). In both S. pyogenes and S. pneumoniae, erythromycin resistance related to a mef gene meant that telithromycin MICs were definitely higher than in erythromycin-susceptible isolates, although telithromycin susceptibility was preserved in all cases. In S. pyogenes, the activity of both 16-membered macrolides and telithromycin against the iMLS(B) strains proved to be dependent on the erm gene involved, being greater against isolates with erm(A).

Viable Newcastle disease vaccine strains in a pharmaceutical dump.

To assess the viability of discarded and buried vaccine strains, we examined vaccines that had been buried for >20 years in an industrial waste dump in the city of Milan, Italy. Viability results showed potential biological risk associated with uncontrolled burial of pharmaceutical industry waste, including some live vaccines.

Neurogenin3 triggers beta-cell differentiation of retinoic acid-derived endoderm cells.

Neurogenin3 is a member of the basic helix-loop-helix ('bHLH') family of transcription factors. It plays a crucial role in the commitment of embryonic endoderm into the pancreatic differentiation programme. This factor is considered to act upstream of a cascade of other transcription factors, leading to the fully differentiated endocrine phenotype. Direct observation of the sequential activation of these factors starting from Neurogenin3 had never been demonstrated. By using retinoic acid-derived-endoderm F9 cells as a model, the present study indicates that the ectopic expression of Neurogenin3 is able to start the differentiation pathway of endocrine pancreas. Neurogenin3 triggers the expression of several pancreatic transcription factors following a well defined temporal activation sequence. By reverse transcriptase PCR, immunohistochemistry and RIA, it is shown that stable transfected cells are able to form embryod bodies that produce insulin in response to glucose stimulation. This is the first report of a differentiation event induced by the ectopic expression of a transcription factor in embryonic pluripotent stem cells.