Publications by authors named "Stefania Stefani"

145 Publications

Challenges in the management of chronic wound infections.

J Glob Antimicrob Resist 2021 Jun 15. Epub 2021 Jun 15.

Maria Cecilia Hospital, Cotignola, Italy.

Objective: Chronic wound infections may delay the healing process and are responsible for a significant burden for the healthcare system. Since inappropriate management may commonly occur in the management of these patients, this review aims to provide a practical guide underlining actions to avoid in the management of chronic wound infections.

Methods: We performed a systematic review of the literature available on PubMed in the last 10 years, identifying studies about the management of patients with chronic wound infections. A panel of experts discussed about the potential malpractices in this area. A list of Don'ts including the main actions to be avoided was drawn up through the Choosing Wisely methodology.

Results: In this review we proposed a list of actions to avoid for an optimal management of these patients. The adequate wound bed preparation and the wound antisepsis should be combined, because the absence of one of them lead to delayed healing and higher risk of wound complications. Moreover, avoiding inappropriate use of systemic antibiotics is an important point because of the risk of selection of multidrug resistant organisms and antibiotic-related adverse events.

Conclusions: A multidisciplinary team of experts in different fields (surgeon, infectious disease expert, microbiologist, pharmacologist, geriatrician) is required for an optimal management of chronic wound infections. The implementation of this approach may be useful to improve the management of patients with chronic wound infections.
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http://dx.doi.org/10.1016/j.jgar.2021.05.010DOI Listing
June 2021

Combination of aztreonam, ceftazidime-avibactam and amikacin in the treatment of VIM-1 Pseudomonas aeruginosa ST235 osteomyelitis.

Int J Infect Dis 2021 Jun 3. Epub 2021 Jun 3.

Infectious Diseases Section, Department of Medicine and Surgery, University of Insubria, Varese, Italy.

We describe a challenging case of a patient with MBL-producing Pseudomonas aeruginosa sternal osteomyelitis following aortic valve replacement with biological prosthesis. The strain exhibited a multidrug-resistance phenotype carrying the bla1 gene and belonged to the high-risk clone sequence type ST235. The patient was successfully treated with surgical debridement plus antibiotic therapy with ceftazidime/avibactam, aztreonam, and amikacin. Time kill curves showed that this triple antibiotic combination at 1 X MIC was strongly synergic after 8 hours, achieving 99.9% killing, and maintaining this until 48 hours.
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http://dx.doi.org/10.1016/j.ijid.2021.05.085DOI Listing
June 2021

Resistance to Echinocandins Complicates a Case of Bloodstream Infection: A Case Report.

J Fungi (Basel) 2021 May 21;7(6). Epub 2021 May 21.

U.O.C. Laboratory Analysis Unit, A.O.U. Policlinico-San Marco, 95123 Catania, Italy.

Invasive candidiasis is known to be one of the most common healthcare-associated complications and is caused by several species. First-line drugs, particularly echinocandins, are effective, but there are increasing reports of resistance to these molecules, though rarely related to . Even though the rate of echinocandins resistance remains low (<3%), sporadic cases are emerging. Here, we present a case of bloodstream infection by a pan-echinocandin-resistant affecting a critically ill patient, who died in an intensive care unit following therapeutic failure and multiple organ dysfunction syndrome. This case highlights the need to suspect pan-echinocandin resistance in patients with prolonged echinocandin exposure, particularly in the presence of urinary tract colonization. Our study shows the importance of sequencing to predict therapeutic failure in patients treated with echinocandins and persistent candidemia.
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http://dx.doi.org/10.3390/jof7060405DOI Listing
May 2021

In vitro activity of cefiderocol and comparators against isolates of Gram-negative pathogens from a range of infection sources: SIDERO-WT-2014-2018 studies in Italy.

J Glob Antimicrob Resist 2021 Jun 18;25:390-398. Epub 2021 May 18.

Department of Biomedical and Biotechnological Sciences, Università di Catania, Via Santa Sofia 97, I-95123 Catania, Italy.

Objectives: Antimicrobial resistance, particularly carbapenem resistance, in Gram-negative pathogens poses a significant healthcare threat. Carbapenem resistance rates in Italy are among the highest in Europe. We report the in vitro activity of cefiderocol, a novel siderophore cephalosporin, and comparator antibiotics against Gram-negative isolates from Italy as part of the SIDERO-WT studies.

Methods: Isolates were collected between 2014 and 2018. Minimum inhibitory concentrations (MICs) were determined using International Organization for Standardization and EUCAST guidelines. Antimicrobial susceptibilities were interpreted using EUCAST breakpoints; pharmacodynamic/pharmacokinetic breakpoints were used if EUCAST breakpoints were not specified.

Results: The 2472 isolates [1545 (62.5%) Enterobacterales and 927 (37.5%) non-fermenters] represented a range of infection sources, including nosocomial pneumonia (902; 36.5%), complicated urinary tract infection (374; 15.1%), bloodstream infection (596; 24.1%), complicated intra-abdominal infection (257; 10.4%) and other infection sources (343; 13.9%). Cefiderocol was active against the majority of isolates, regardless of infection source (susceptibility, 94.2-97.3%). A high proportion of non-fermenters (97.6%) and Enterobacterales (95.6%) were cefiderocol-susceptible, although susceptibility was lower in Klebsiella pneumoniae (88.1%). Susceptibility to cefiderocol was significantly (P < 0.01) greater than comparators overall (96.4% vs. 71.3-81.6%) and in non-fermenters (97.6% vs. 44.3-90.3%) across infection sources. Overall 612/2472 isolates (24.8%) were meropenem-resistant (MIC > 8 mg/L), comprising 516/927 (55.7%) non-fermenters and 96/1545 (6.2%) Enterobacterales. Cefiderocol (499/516; 96.7%) activity was greater than colistin (440/516; 85.3%), ceftazidime/avibactam (123/516; 23.8%) and ceftolozane/tazobactam (89/516; 17.2%) in meropenem-resistant non-fermenter isolates.

Conclusion: Susceptibility to cefiderocol was significantly greater than meropenem, colistin, ceftazidime/avibactam and ceftolozane/tazobactam overall, regardless of infection source.
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http://dx.doi.org/10.1016/j.jgar.2021.04.019DOI Listing
June 2021

Diagnostic stewardship based on patient profiles: differential approaches in acute versus chronic infectious syndromes.

Expert Rev Anti Infect Ther 2021 May 14:1-11. Epub 2021 May 14.

Department of Public Health and Infectious Diseases, Sapienza University of Rome, Rome, Italy.

: New diagnostics may be useful in clinical practice, especially in contexts of high prevalence of multidrug-resistant organisms (MDRO). However, misuse of diagnostic tools may lead to increased costs and worse patient outcome. Conventional and new techniques should be appropriately positioned in diagnostic algorithms to guide an appropriate use of antimicrobial therapy.: A panel of experts identified 4 main areas in which the implementation of diagnostic stewardship is needed. Among chronic infections, bone and prosthetic joint infections and subacute-chronic intravascular infections and endocarditis represent common challenges for clinicians. Among acute infections, bloodstream infections and community-acquired pneumonia may be associated with high mortality and require appropriate diagnostic approach.: Diagnostic stewardship aims to improve the appropriate use of microbiological diagnostics to guide therapeutic decisions through appropriate and timely diagnostic testing. Here, diagnostic algorithms based on different patient profiles are proposed for chronic and acute clinical syndromes. In each clinical scenario, combining conventional and new diagnostic techniques is crucial to make a rapid and accurate diagnosis and to guide the selection of antimicrobial therapy. Barriers related to the implementation of new rapid diagnostic tools, such as high initial costs, may be overcome through their rational and structured use.
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http://dx.doi.org/10.1080/14787210.2021.1926986DOI Listing
May 2021

Antimicrobial properties of Lactobacillus cell-free supernatants against multidrug-resistant urogenital pathogens.

Microbiologyopen 2021 Feb;10(2):e1173

Department of Biomedical and Biotechnological Sciences, Microbiology Section, University of Catania, Catania, Italy.

The healthy vaginal microbiota is dominated by Lactobacillus spp., which provide an important critical line of defense against pathogens, as well as giving beneficial effects to the host. We characterized L. gasseri 1A-TV, L. fermentum 18A-TV, and L. crispatus 35A-TV, from the vaginal microbiota of healthy premenopausal women, for their potential probiotic activities. The antimicrobial effects of the 3 strains and their combination against clinical urogenital bacteria were evaluated together with the activities of their metabolites produced by cell-free supernatants (CFSs). Their beneficial properties in terms of ability to interfere with vaginal pathogens (co-aggregation, adhesion to HeLa cells, biofilm formation) and antimicrobial activity mediated by CFSs were assessed against multidrug urogenital pathogens (S. agalactiae, E. coli, KPC-producing K. pneumoniae, S. aureus, E. faecium VRE, E. faecalis, P. aeruginosa, P. mirabilis, P. vulgaris, C. albicans, C. glabrata). The Lactobacilli tested exhibited an extraordinary ability to interfere and co-aggregate with urogenital pathogens, except for Candida spp., as well as to adhere to HeLa cells and to produce biofilm in the Lactobacillus combination. Lactobacillus CFSs and their combination revealed a strong bactericidal effect on the multidrug resistant indicator strains tested, except for E. faecium and E. faecalis. The antimicrobial activity was maintained after heat treatment but decreased after enzymatic treatment. All Lactobacilli showed lactic dehydrogenase activity and production of D- and L-lactic acid isomers on Lactobacillus CFSs, while only 1A-TV and 35A-TV released hydrogen peroxide and carried helveticin J and acidocin A bacteriocins. These results suggest that they can be employed as a new vaginal probiotic formulation and bio-therapeutic preparation against urogenital infections. Further, in vivo studies are needed to evaluate human health benefits in clinical situations.
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http://dx.doi.org/10.1002/mbo3.1173DOI Listing
February 2021

Staphylococcus aureus ST228 and ST239 as models for expression studies of diverse markers during osteoblast infection and persistence.

Microbiologyopen 2021 Mar;10(2):e1178

Department of Biomedical and Biotechnological Sciences (BIOMETEC), Medical Molecular Microbiology and Antibiotic Resistance laboratory (MMARLab, University of Catania, Catania, Italy.

The ability of S. aureus to infect bone and osteoblasts is correlated with its incredible virulence armamentarium that can mediate the invasion/internalization process, cytotoxicity, membrane damage, and intracellular persistence. We comparatively analyzed the interaction, persistence, and modulation of expression of selected genes and cell viability in an ex vivo model using human MG-63 osteoblasts of two previously studied and well-characterized S. aureus clinical strains belonging to the ST239-SCCmecIII-t037 and ST228-SCCmecI-t041 clones at 3 h and 24 h post-infection (p.i). S. aureus ATCC12598 ST30-t076 was used as a control strain. Using imaging flow cytometry (IFC), we found that these strains invaded and persisted in MG-63 osteoblasts to different extents. The invasion was evaluated at 3 h p.i and persistence at 24 h p.i., in particular: ATCC12598 internalized in 70% and persisted in 50% of MG-63 cells; ST239-SCCmecIII internalized in 50% and persisted in 45% of MG-63 cells; and ST228-SCCmecI internalized in 30% and persisted in 20% of MG-63 cells. During the infection period, ST239-III exerted significant cytotoxic activity resulting from overexpression of hla and psmA and increased expression of the genes involved in adhesion, probably due to the release and re-entry of bacteria inside MG-63 cells at 24 h p.i. The lower invasiveness of ST228-I was also associated with non-cytotoxic activity inside osteoblasts. This clone was unable to activate sufficient cellular reaction and succumbed inside MG-63 cells. Our findings support the idea of considering new strategies, based on a translational approach-eukaryotic host-pathogen interaction (EHPI)-and to be applied on a large scale, to predict S. aureus /osteoblast interaction and treat bone infections. Such strategies rely on the study of the genetic and biochemical basis of both pathogen and host.
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http://dx.doi.org/10.1002/mbo3.1178DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8087985PMC
March 2021

Internalization in Osteoblast Cells: Mechanisms, Interactions and Biochemical Processes. What Did We Learn from Experimental Models?

Pathogens 2021 Feb 19;10(2). Epub 2021 Feb 19.

Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMARLab), Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95125 Catania, Italy.

Bacterial internalization is a strategy that non-intracellular microorganisms use to escape the host immune system and survive inside the human body. Among bacterial species, showed the ability to interact with and infect osteoblasts, causing osteomyelitis as well as bone and joint infection, while also becoming increasingly resistant to antibiotic therapy and a reservoir of bacteria that can make the infection difficult to cure. Despite being a serious issue in orthopedic surgery, little is known about the mechanisms that allow bacteria to enter and survive inside the osteoblasts, due to the lack of consistent experimental models. In this review, we describe the current knowledge about internalization mechanisms and various aspects of the interaction between bacteria and osteoblasts (e.g., best experimental conditions, bacteria-induced damages and immune system response), focusing on studies performed using the MG-63 osteoblastic cell line, the best traditional (2D) model for the study of this phenomenon to date. At the same time, as it has been widely demonstrated that 2D culture systems are not completely indicative of the dynamic environment in vivo, and more recent 3D models-representative of bone infection-have also been investigated.
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http://dx.doi.org/10.3390/pathogens10020239DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7922271PMC
February 2021

Testing Surgical Face Masks in an Emergency Context: The Experience of Italian Laboratories during the COVID-19 Pandemic Crisis.

Int J Environ Res Public Health 2021 02 4;18(4). Epub 2021 Feb 4.

Presidio Tecnico/Scientifico di Ateneo per l'Emergenza COVID-19, Centro Servizi Metrologici e Tecnologici Avanzati, Università di Napoli Federico II, 80124 Naples, Italy.

The first wave of the COVID-19 pandemic brought about a broader use of masks by both professionals and the general population. This resulted in a severe worldwide shortage of devices and the need to increase import and activate production of safe and effective surgical masks at the national level. In order to support the demand for testing surgical masks in the Italian context, Universities provided their contribution by setting up laboratories for testing mask performance before releasing products into the national market. This paper reports the effort of seven Italian university laboratories who set up facilities for testing face masks during the emergency period of the COVID-19 pandemic. Measurement set-ups were built, adapting the methods specified in the EN 14683:2019+AC. Data on differential pressure (DP) and bacterial filtration efficiency (BFE) of 120 masks, including different materials and designs, were collected over three months. More than 60% of the masks satisfied requirements for DP and BFE set by the standard. Masks made of nonwoven polypropylene with at least three layers (spunbonded-meltblown-spunbonded) showed the best results, ensuring both good breathability and high filtration efficiency. The majority of the masks created with alternative materials and designs did not comply with both standard requirements, resulting in suitability only as community masks. The effective partnering between universities and industries to meet a public need in an emergency context represented a fruitful example of the so-called university "third-mission".
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http://dx.doi.org/10.3390/ijerph18041462DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7915703PMC
February 2021

Genomic Characterization of a New Biofilm-Forming and Adhesive ST398 Human-Adapted MSSA Lineage Causing Septic Knee Arthritis Following Surgical Reconstruction.

Microorganisms 2021 Feb 2;9(2). Epub 2021 Feb 2.

Department of Biomedical and Biotechnological Sciences, University of Catania, 95123 Catania, Italy.

Methicillin-susceptible (MSSA) and methicillin-resistant (MRSA) is a pathogen commonly found in bone and joint infections, including septic arthritis. virulence and the frailty of affected patients can cause several complications; a prompt and specific antibiotic treatment can positively affect the outcome of patients. We carried out an in-depth genomic characterization by Illumina whole genome sequencing and bioinformatics of two biofilm-producing M1 and M2 ST398 MSSA causing septic knee arthritis not-responding to antimicrobial therapy. The strains were characterized for antibiotic resistance, biofilm and adhesive properties as well as genomics, single nucleotide polymorphism phylogeny, resistomics and virulomics. Our results showed that M1 and M2 MSSA were ST398-t1451-I-Cap5, susceptible to cefoxitin and resistant to erythromycin and clindamycin, traits consistent with the lack of the SCC-locus and the presence of the sole Z and T. Furthermore, M1 and M2 were biofilm-producing and largely potentially adhesive strains, as indicated by the adhesion gene profile. Our data characterized a new human-adapted ST398 MSSA lineage, representing a "fusion" between the human-animal independent ST398 and the Livestock Associated (LA) ST398 lineages, forming biofilm and genomically predicted high adhesive, characterized by different genomic adaptation conferring a great ability to adhere to the host's extracellular matrix causing septic knee arthritis.
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http://dx.doi.org/10.3390/microorganisms9020305DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7913009PMC
February 2021

Different Modulatory Effects of Four Methicillin-Resistant Clones on MG-63 Osteoblast-Like Cells.

Biomolecules 2021 01 7;11(1). Epub 2021 Jan 7.

Department of Biomedical and Biotechnological Sciences (BIOMETEC), University of Catania, 95125 Catania, Italy.

is a Gram-positive bacterium responsible for a variety of mild to life-threatening infections including bone infections such as osteomyelitis. This bacterium is able to invade and persist within non-professional phagocytic cells such as osteoblasts. In the present study, four different strains, namely, 2SA-ST239-III (ST239), 5SA-ST5-II (ST5), 10SA-ST228-I (ST228), and 14SA-ST22-IVh (ST22), were tested for their ability to modulate cell viability in MG-63 osteoblast-like cells following successful invasion and persistence. Methicillin-sensitive (MSSA) ATCC-12598-ST30 (ST30) was used as control strain. Despite being proven that ST30, ST239, and ST22 have a similar ability to internalize and persist in MG-63 osteoblast-like cells under our experimental conditions, we demonstrated that the observed decrease in cell viability was due to the different behavior of the considered strains, rather than the number of intracellular bacteria. We focused our attention on different biochemical cell functions related to inflammation, cell metabolism, and oxidative stress during osteoblast infections. We were able to show the following: (1) ST30 and ST239 were the only two clones able to persist and maintain their number in the hostile environment of the cell during the entire period of infection; (2) ST239 was the only clone able to significantly increase gene expression (3 and 24 h post-infection (p.i.)) and protein secretion (24 h p.i.) of both interleukin-6 (IL-6) and tumor necrosis factor alpha (TNF-α) in MG-63 osteoblast-like cells; (3) the same clone determined a significant up-regulation of the transforming growth factorbeta 1 (TGF-β1) and of the metabolic marker glyceraldehyde 3-phosphate dehydrogenase (GAPDH) mRNAs at 24 h p.i.; and (4) neither the MSSA nor the four methicillin-resistant (MRSA) strains induced oxidative stress phenomena in MG-63 cells, although a high degree of variability was observed for the different clones with regard to the expression pattern of nuclear factor E2-related factor 2 (Nrf2) and its downstream gene heme oxygenase 1 (HO-1) activation. Our results may pave the way for an approach to -induced damage, moving towards individualized therapeutic strategies that take into account the differences between MSSA and MRSA as well as the distinctive features of the different clones. This approach is based on a change of paradigm in antibiotic therapy involving a case-based use of molecules able to counteract pro-inflammatory cytokines activity such as selective cytokine signaling inhibitors (IL-6, TNF-α).
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http://dx.doi.org/10.3390/biom11010072DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7825699PMC
January 2021

In Vitro Activity of Dalbavancin against Refractory Multidrug-Resistant (MDR) Isolates.

Antibiotics (Basel) 2020 Dec 3;9(12). Epub 2020 Dec 3.

Department of Biomedical and Biotechnological Sciences (BIOMETEC)-Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMARLab), University of Catania, 95123 Catania, Italy.

The high prevalence of methicillin-resistant (MRSA) infections, always treated with vancomycin and daptomycin, has led to the emergence of vancomycin-intermediate (VISA), heteroresistant vancomycin-intermediate (hVISA) and daptomycin non-susceptible (DNS) . Even if glycopeptides and daptomycin remain the keystone for treatment of resistant , the need for alternative therapies that target MRSA has now become imperative. The in vitro antibacterial and bactericidal activity of dalbavancin was evaluated against clinically relevant showing raised antibiotic resistance levels, from methicillin-susceptible to Multidrug-Resistant (MDR) MRSA, including hVISA, DNS and rifampicin-resistant (RIF-R) strains. A total of 124 strains were tested for dalbavancin susceptibility, by the broth microdilution method. Two VISA and 2 hVISA reference strains, as well as a vancomycin-resistant (VRSA) reference strain and a methicillin-susceptible (MSSA) reference strain, were included as controls. Time-kill curves were assayed to assess bactericidal activity. Dalbavancin demonstrated excellent in vitro antibacterial and bactericidal activity against all resistance classes, including hVISA and DNS isolates. The RIF-R strains showed the highest percentage of isolates with non-susceptibility, reflecting the correlation between B mutations and VISA/hVISA emergence. Our observations suggest that dalbavancin can be considered as an effective alternative for the management of severe MRSA infections also sustained by refractory phenotypes.
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http://dx.doi.org/10.3390/antibiotics9120865DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7761838PMC
December 2020

Combined Effects of the 2039 A/G and -29 G/A Polymorphisms on Male Reproductive Parameters.

World J Mens Health 2020 Sep 28. Epub 2020 Sep 28.

Department of Clinical and Experimental Medicine, University of Catania, Catania, Italy.

Purpose: The aim of this study was to evaluate the combined effect of 2039 A/G and -29 G/A single nucleotide polymorphisms (SNPs) on the male reproductive function in a cohort of Sicilian men.

Materials And Methods: One-hundred thirty Sicilian men were enrolled and underwent blood withdrawal for hormone measurement and 2039 A/G and -29 G/A SNP genotyping, testicular volume evaluation by ultrasound scan, and semen analysis. A meta-analysis of the -29 G/A SNP, evaluated in a previous study of the Sicilian population was done.

Results: No genotype of the 2039 A/G SNP correlated with serum follicle-stimulating hormone (FSH) levels, testicular volume, sperm concentration, and total sperm count. In contrast, normozoospermic men with -29 GG and -29 GA genotypes had significantly lower sperm concentrations compared to men with the -29 AA genotype. The other sperm parameters did not show any significant difference. The meta-analysis showed no significant difference in serum FSH levels, testicular volume, sperm concentration, and total sperm count between -29 GG and -29 AA in Sicilian men. No difference was found even when the two SNPs were evaluated in combination. However, this combination was present, as expected, only in a low proportion (3.8%) of the men studied.

Conclusions: The SNPs 2039 A/G and -29 G/A in combination did not seem to have any effect on male reproductive function in a cohort of Sicilian men. The effect of these SNPs has only been studied in granulosa cells so far. Further studies on their role in Sertoli cells are needed.
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http://dx.doi.org/10.5534/wjmh.200070DOI Listing
September 2020

SARS-CoV-2 diagnostics: Some reflections on current assays.

Diagn Microbiol Infect Dis 2021 Feb 7;99(2):115237. Epub 2020 Oct 7.

Department of Translational Medicine and New Technologies in Medicine and Surgery, Retrovirus Centre, University of Pisa, "Clinical Virology Service," Pisa University Hospital, Pisa, Italy.

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http://dx.doi.org/10.1016/j.diagmicrobio.2020.115237DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7538382PMC
February 2021

Gold standard susceptibility testing of fosfomycin in Staphylococcus aureus and Enterobacterales using a new agar dilution panel®.

J Glob Antimicrob Resist 2020 12 27;23:334-337. Epub 2020 Oct 27.

Department of Biomedical and Biotechnological Sciences, section of Microbiology, University of Catania, Italy.

Objectives: Many clinical laboratories have difficulty in routinely performing in vitro fosfomycin susceptibility testing using the agar dilution (AD) method, considered to be the gold standard method. The objective of our work was to evaluate a rapid commercial fosfomycin agar dilution panel against clinical Staphylococcus aureus and Enterobacterales strains, in two different centres located in Italy and in the UK.

Methods: A total of 99 Enterobacterales (mostly Escherichia coli and Klebsiella pneumoniae) and 80 S. aureus clinical isolates was used to evaluate the commercial device, a 12-well panel containing fosfomycin incorporated into CA-MH agar supplemented with 25mg/L of glucose-6-phosphate (Liofilchem S.r.l., Roseto degli Abruzzi, Italy). Testing was performed in two centres (Italy and UK) and kit results were compared against the gold standard in-house AD MIC method.

Results: According to the EUCAST breakpoints, fosfomycin inhibited 61% of the S. aureus strains, and 76% of the Enterobacterales isolates tested by the AD reference method. There was a Categorical Agreement (CA) of 100% and an Essential Agreement (EA) of 91.25% for S. aureus; while the Enterobacterales strains showed a CA of 94% and an EA of 97%. No evaluation errors were observed among S. aureus, while 5% Major Error and 1% Very Major Error were observed for the Enterobacterales.

Conclusions: Our results confirmed the feasibility of determining fosfomycin susceptibility using a commercial AD panel as a routine substitution for the AD test. The few differences observed were only in strains with MICs around the breakpoint used.
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http://dx.doi.org/10.1016/j.jgar.2020.08.025DOI Listing
December 2020

In vitro fosfomycin study on concordance of susceptibility testing methods against ESBL and carbapenem-resistant Enterobacteriaceae.

J Glob Antimicrob Resist 2020 12 9;23:286-289. Epub 2020 Oct 9.

Department of Biomedical and Biotechnological Sciences, Section of Microbiology, University of Catania, Italy. Electronic address:

Objectives: The increasing emergence and diffusion of multidrug-resistant (MDR) pathogenic bacteria, both in hospital and community settings, is inducing clinicians to reconsider old antibiotics, such as fosfomycin, to overcome the difficulties posed by these microorganisms. Recent studies have reported good in vitro activity of fosfomycin against extended spectrum ß-lactamases (ESBL) and carbapenem-resistant Enterobacteriaceae. The aim of this study was to assess thein vitro activity of fosfomycin by different methods against 120 clinical MDR isolates.

Methods: Fosfomycin minimum inhibitory concentrations were determined using the agar dilution reference method (AD), gradient test (GT), broth microdilution method (BMD), according to CLSI recommendations, and automated systems (VITEK 2 and BD Phoenix) against 85 carbapenem-resistant Klebsiella pneumoniae and 35 ESBL-producing Escherichia coli. Agreement and discrepancies between the evaluated methods and the reference method were calculated.

Results: Fosfomycin showed very good activity against ESBL-producing E. coli (88.6%). Excellent agreement (100%) between the three (AD, BMD and GT) susceptibility methods was found for E. coli. No major errors were observed. The fosfomycin resistance rate ranged from 24% (KPC-producing) to 100% (NDM-OXA-48 co-producing) K. pneumoniae. For all carbapenem-resistant K. pneumoniae strains, categorical agreement was >90% for all methods except for VITEK 2, which was 84%.

Conclusions: When ESBL E. coli isolates are found to be susceptible to fosfomycin with automated systems, it is not necessary to verify these results with the AD reference method; while for resistant strains, the GT can be used. In cases of KPC K. pneumoniae resistant to fosfomycin, the AD method is the only reference method.
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http://dx.doi.org/10.1016/j.jgar.2020.09.022DOI Listing
December 2020

Genomic and Long-Term Transcriptomic Imprints Related to the Daptomycin Mechanism of Action Occurring in Daptomycin- and Methicillin-Resistant Under Daptomycin Exposure.

Front Microbiol 2020 14;11:1893. Epub 2020 Aug 14.

Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy.

Daptomycin (DAP) is one of the last-resort treatments for heterogeneous vancomycin-intermediate (hVISA) and vancomycin-intermediate (VISA) infections. DAP resistance (DAP-R) is multifactorial and mainly related to cell-envelope modifications caused by single-nucleotide polymorphisms and/or modulation mechanisms of transcription emerging as result of a self-defense process in response to DAP exposure. Nevertheless, the role of these adaptations remains unclear. We aim to investigate the comparative genomics and late post-exponential growth-phase transcriptomics of two DAP-resistant/DAP-susceptible (DAP) methicillin-resistant (MRSA) clinical strain pairs to focalize the genomic and long-term transcriptomic fingerprinting and adaptations related to the DAP mechanism of action acquired under DAP pressure using Illumina whole-genome sequencing (WGS), RNA-seq, bioinformatics, and real-time qPCR validation. Comparative genomics revealed that membrane protein and transcriptional regulator coding genes emerged as shared functional coding-gene clusters harboring mutational events related to the DAP-R onset in a strain-dependent manner. Pairwise transcriptomic enrichment analysis highlighted common and strain pair-dependent Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways, whereas DAP double-pair cross-filtering returned 53 differentially expressed genes (DEGs). A multifactorial long-term transcriptomic-network characterized DAP MRSA includes alterations in (i) peptidoglycan biosynthesis, cell division, and cell-membrane (CM) organization genes, as well as a B/S autolysin genes; (ii) 2 involved in fermentative metabolism; (iii) CM-potential perturbation genes; and (iv) oxidative and heat/cold stress response-related genes. Moreover, a D-alanyl-D-alanine decrease in cell-wall muropeptide characterized DAP/glycopeptide cross-reduced susceptibility mechanisms in DAP MRSA. Our data provide a snapshot of DAP MRSA genomic and long-term transcriptome signatures related to the DAP mechanism of action (MOA) evidencing that a complex network of genomic changes and transcriptomic adaptations is required to acquire DAP-R.
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http://dx.doi.org/10.3389/fmicb.2020.01893DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7456847PMC
August 2020

Potential Associations Among Alteration of Salivary miRNAs, Saliva Microbiome Structure, and Cognitive Impairments in Autistic Children.

Int J Mol Sci 2020 Aug 27;21(17). Epub 2020 Aug 27.

Department of Biomedical and Biotechnological Sciences, Section of Biology and Genetics G. Sichel, University of Catania, 95123 Catania, Italy.

Recent evidence has demonstrated that salivary molecules, as well as bacterial populations, can be perturbed by several pathological conditions, including neuro-psychiatric diseases. This relationship between brain functionality and saliva composition could be exploited to unveil new pathological mechanisms of elusive diseases, such as Autistic Spectrum Disorder (ASD). We performed a combined approach of miRNA expression profiling by NanoString technology, followed by validation experiments in qPCR, and 16S rRNA microbiome analysis on saliva from 53 ASD and 27 neurologically unaffected control (NUC) children. MiR-29a-3p and miR-141-3p were upregulated, while miR-16-5p, let-7b-5p, and miR-451a were downregulated in ASD compared to NUCs. Microbiome analysis on the same subjects revealed that , and increased their abundance in ASD patients, while and decreased. Variations of both miRNAs and microbes were statistically associated to different neuropsychological scores related to anomalies in social interaction and communication. Among miRNA/bacteria associations, the most relevant was the negative correlation between salivary miR-141-3p expression and abundance. MiRNA and microbiome dysregulations found in the saliva of ASD children are potentially associated with cognitive impairments of the subjects. Furthermore, a potential cross-talking between circulating miRNAs and resident bacteria could occur in saliva of ASD.
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http://dx.doi.org/10.3390/ijms21176203DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7504581PMC
August 2020

Sensitivity assessment of droplet digital PCR for SARS-CoV-2 detection.

Int J Mol Med 2020 Sep 13;46(3):957-964. Epub 2020 Jul 13.

Department of Biomedical and Biotechnological Sciences, Section of Microbiology, University of Catania, I‑95123 Catania, Italy.

Reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) is the gold standard method for the diagnosis of COVID‑19 infection. Due to pre‑analytical and technical limitations, samples with low viral load are often misdiagnosed as false‑negative samples. Therefore, it is important to evaluate other strategies able to overcome the limits of RT‑qPCR. Blinded swab samples from two individuals diagnosed positive and negative for COVID‑19 were analyzed by droplet digital PCR (ddPCR) and RT‑qPCR in order to assess the sensitivity of both methods. Intercalation chemistries and a World Health Organization (WHO)/Center for Disease Control and Prevention (CDC)‑approved probe for the SARS‑CoV‑2 N gene were used. SYBR‑Green RT‑qPCR is not able to diagnose as positive samples with low viral load, while, TaqMan Probe RT‑qPCR gave positive signals at very late Ct values. On the contrary, ddPCR showed higher sensitivity rate compared to RT‑qPCR and both EvaGreen and probe ddPCR were able to recognize the sample with low viral load as positive even at 10‑fold diluted concentration. In conclusion, ddPCR shows higher sensitivity and specificity compared to RT‑qPCR for the diagnosis of COVID‑19 infection in false‑negative samples with low viral load. Therefore, ddPCR is strongly recommended in clinical practice for the diagnosis of COVID‑19 and the follow‑up of positive patients until complete remission.
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http://dx.doi.org/10.3892/ijmm.2020.4673DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7388836PMC
September 2020

The 2039 A/G FSH receptor gene polymorphism influences glucose metabolism in healthy men.

Endocrine 2020 12 17;70(3):629-634. Epub 2020 Jul 17.

Department of Clinical and Experimental Medicine, University of Catania, Catania, Italy.

Objective: To assess the role of c. 2039 A/G (p. Asp680Ser) (rs6166) and c. -29 G/A (rs1394205) follicle-stimulating hormone receptor (FSHR) gene single nucleotide polymorphisms (SNPs) in a cohort of healthy men.

Methods: One-hundred twenty-seven healthy men underwent evaluation of the anthropometric parameters, assessment of metabolic and lipid profile, measurement FSH serum levels, and genotyping of both the aforementioned FSHR SNPs. Data grouped according to the FSHR rs6166 or rs1394205 genotypes underwent to statistical analysis.

Main Results: The three groups of men for each FSHR SNP did not differ statistically significantly for body mass index and serum FSH levels. As for FSHR rs6166 SNP, glucose levels were significantly lower in men with the GG genotype compared with those with the AA genotype. Men with AG had lower insulin levels and HOMA index values compared with those carrying the genotype AA (p < 0.05). The GG group showed a negative correlation between serum FSH levels and insulin and between serum FSH levels and HOMA index (p < 0.05). In contrast, men grouped according to the FSHR rs1394205 genotype showed no significant difference in blood glucose, serum insulin levels, and HOMA index. The AG group showed a negative correlation between FSH insulin and between serum FSH levels and HOMA index (p < 0.05).

Conclusions: Men with the genotype GG of the FSHR rs6166 SNP have lower blood glucose levels than those with the AA genotype. Their FSH levels inversely correlated with insulin and HOMA index. In contrast, the genotype FSHR rs6166 A/G did not reveal any role of FSH on glucose metabolism in healthy men. The inverse relationship between FSH and insulin or HOMA index in the group with the genotype GG of the FSHR rs6166 SNP suggests a possible cross-talk between FSH and insulin.
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http://dx.doi.org/10.1007/s12020-020-02420-3DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7674314PMC
December 2020

Fluoroquinolone Metalloantibiotics: A Promising Approach against Methicillin-Resistant .

Int J Environ Res Public Health 2020 04 30;17(9). Epub 2020 Apr 30.

REQUIMTE-LAQV (Rede de Química e Tecnologia - Laboratório Associado para a Química Verde), Departamento de Química e Bioquímica da Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre, s/n, 4169-007 Porto, Portugal.

Fluoroquinolones (FQs) are antibiotics commonly used in clinical practice, although nowadays they are becoming ineffective due to the emergence of several mechanisms of resistance in most bacteria. The complexation of FQs with divalent metal ions and phenanthroline (phen) is a possible approach to circumvent antimicrobial resistance, since it forms very stable complexes known as metalloantibiotics. This work is aimed at determining the antimicrobial activity of metalloantibiotics of Cu(II)FQphen against a panel of multidrug‑resistant (MDR) clinical isolates and to clarify their mechanism of action. Minimum inhibitory concentrations (MICs) were determined against MDR isolates of and methicillin-resistant (MRSA). Metalloantibiotics showed improved antimicrobial activity against several clinical isolates, especially MRSA. Synergistic activity was evaluated in combination with ciprofloxacin and ampicillin by the disk diffusion and checkerboard methods. Synergistic and additive effects were shown against MRSA isolates. The mechanism of action was studied though enzymatic assays and atomic force microscopy (AFM) experiments. The results indicate a similar mechanism of action for FQs and metalloantibiotics. In summary, metalloantibiotics seem to be an effective alternative to pure FQs against MRSA. The results obtained in this work open the way to the screening of metalloantibiotics against other Gram‑positive bacteria.
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http://dx.doi.org/10.3390/ijerph17093127DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7246690PMC
April 2020

Detection of methicillin-resistant Staphylococcus aureus persistence in osteoblasts using imaging flow cytometry.

Microbiologyopen 2020 05 1;9(5):e1017. Epub 2020 Apr 1.

Department of Biomedical and Biotechnological Sciences (BIOMETEC), Medical Molecular Microbiology and Antibiotic Resistance Laboratory (MMARLab), University of Catania, Catania, Italy.

Methicillin-resistant S. aureus has been reported as the main pathogen involved in chronic infections, osteomyelitis, and prosthetic joint infections. The host/pathogen interaction is dynamic and requires several changes to promote bacterial survival. Here, we focused on the internalization and persistence behavior of well-characterized Staphylococcus aureus invasive strains belonging to the main ST-MRSA-SCCmec clones. To overcome the limitations of the cell culture method, we comparatively analyzed the ability of internalization within human MG-63 osteoblasts with imaging flow cytometry (IFC). After evaluation by cell culture assay, the MRSA clones in the study were all able to readily internalize at 3h postinfection, the persistence of intracellular bacteria was evaluated at 24h both by routine cell culture and IFC assay, after vancomycin-BODIPY staining. A statistical difference of persistence was found in ST5-SCCmecII (26.59%), ST228-SCCmecI (20.25%), ST8-SCCmecIV (19.52%), ST239-SCCmecIII (47.82%), and ST22-SCCmecIVh (50.55%) showing the same ability to internalize as ATCC12598 (51%), the invasive isolate used as control strain for invasion and persistence assays. We demonstrated that the intracellular persistence process depends on the total number of infected cells. Comparing our data obtained by IFC with those of the cell culture assay, we obtained greater reproducibility rates and a number of intracellular bacteria, with the advantage of analyzing live host cells. Moreover, with some limitations related to the lack of whole-genome sequencing analysis, we validated the different proclivities to persist in the main Italian HA-MRSA invasive isolates and our results highlighted the heterogeneity of the different clones to persist during cell infection.
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http://dx.doi.org/10.1002/mbo3.1017DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7221431PMC
May 2020

COVID-19 Therapeutic and Prevention.

Int J Antimicrob Agents 2020 04 7;55(4):105937. Epub 2020 Mar 7.

MEФI, IRD, Aix Marseille Univ, AP-HM, Marseille, France; IHU-Méditerranée Infection, Marseille, France.

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http://dx.doi.org/10.1016/j.ijantimicag.2020.105937DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7135736PMC
April 2020

Antimicrobial stewardship in patients with acute bacterial skin and skin-structure infections: An international Delphi consensus.

J Glob Antimicrob Resist 2020 09 14;22:296-301. Epub 2020 Feb 14.

Infectious Diseases Unit, Cisanello Hospital, AOUP, Department of Clinical and Experimental Medicine, University of Pisa, 56100 Pisa, Italy. Electronic address:

Objectives: The aim of this survey was to identify a set of actions aimed to improve the diagnosis and management of acute bacterial skin and skin-structure infections (ABSSSIs) and the implementation of some principles of antimicrobial stewardship (AMS) in this setting.

Methods: A list of 76 statements for which there was a lack of clarity were generated by an expert panel and were validated by a group of experts. The questionnaire was administered to 112 experts in infectious diseases or microbiology. Participants were asked to vote on a list of statements. An agreement threshold of 66% was required to reach consensus.

Results: Overall, 57 responders participated in the survey. Positive consensus was reached on the fact that ABSSSIs represent a significant cause of infection in the emergency department, are frequently associated with increased hospital stay and are mainly caused by Staphylococcus aureus. The panellists strongly supported collection of samples from purulent infections by needle aspiration as well as collection of blood cultures in the presence of signs/symptoms of systemic infection. The importance of source control and prompt adequate microbiological documentation, the objective to reduce the length of hospital stay, the choice of a narrow-spectrum antibiotic and the role of new therapeutic options (e.g. long-acting drugs) in improving compliance also reached a positive consensus.

Conclusion: This Delphi survey provides useful indicators for the implementation of AMS principles in the clinical management of ABSSSI and offers interesting elements of discussion about the barriers existing in Europe for optimal implementation of AMS programmes.
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http://dx.doi.org/10.1016/j.jgar.2020.02.002DOI Listing
September 2020

Acute wound infections management: the 'Don'ts' from a multidisciplinary expert panel.

Expert Rev Anti Infect Ther 2020 03 17;18(3):231-240. Epub 2020 Feb 17.

Infectious Diseases Unit, Azienda Ospedaliera Universitaria Pisana, Pisa, Italy.

: The management of acute wounds may be affected by malpractices leading to poor outcome, prolonged hospital stay and inappropriate use of antibiotic therapy.: Acute wound infections are represented by surgical site and post-traumatic infections. The aim of this expert opinion is to identify a list of inadvisable actions and to provide a guide for an optimal management of acute wound infections. A literature search using Pubmed/MEDLINE database was performed. Articles pertaining to areas covered published until December 2019 were selected. We identified the most common malpractices in this setting and, using the Choosing Wisely methodology, we proposed a list of "Don'ts" for an easy use in clinical practice.: Malpractices may occur from the surgical prophylaxis to the discharge of patient. A prolonged surgical prophylaxis, the underestimation of signs and symptoms, the omission of source control, the inappropriate collection of wound swab, the improper use of clinical microbiology and pharmacology, the lack of hygiene measures and the delay of discharge are all factors that may lead to unfavorable outcome. A multidisciplinary approach is needed to optimally manage these patients. The "Don'ts" refer to all professional figures involved in the management of patients with acute wound infections.
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http://dx.doi.org/10.1080/14787210.2020.1726740DOI Listing
March 2020

COL sRNA Signatures: Computational Comparative Identification and Biological Targets.

Front Microbiol 2019 17;10:3075. Epub 2020 Jan 17.

Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy.

Multidrug-Resistant (MDR) and Extensively Drug Resistant (XDR) () represent a serious cause of healthcare-associated infections worldwide. Currently, the available treatment options are very restricted and colistin-based therapies are last-line treatments of these infections, even though colistin resistant (COL) have rarely been isolated yet. In bacteria, small non-coding RNAs (sRNAs) have been implicated in regulatory pathways of different biological functions, however, no knowledge exists about the sRNA role on the biological adaptation in COL. Our study investigated two Italian XDR isogenic colistin-susceptible/resistant (COL) strain-pairs to discover new sRNA signatures. Comparative sRNA transcriptome (sRNAome) analyses were carried out by Illumina RNA-seq using both a Tru-Seq and a Short Insert library, whilst ATCC 17978 and ACICU Reference Genome assembly, mapping, annotation and statistically significant differential expression (-value ≤ 0.01) of the raw reads were performed by the Rockhopper tool. A computational filtering, sorting only similarly statistically significant differentially expressed (DE) sRNAs mapping on the same gene in both COL isolates was conducted. COL vs. COL sRNAome, analyzed integrating the DE sRNAs obtained from the two different libraries, revealed some statistically significant DE sRNAs in COL. In detail, we found: (i) two different under-expressed -acting sRNAs (sRNA and sRNA) mapping in antisense orientation the 16S rRNA gene A1S_r01, (ii) one under-expressed -acting sRNA (sRNA) targeting the A1S_2505 gene (hypothetical protein), (iii) one under-expressed microRNA-size small RNA fragment (sRNA) and its pre-microsRNA targeting the A1S_0501 gene (hypothetical protein), (iv) as well as an over-expressed microRNA-size small RNA fragment (sRNA) and its pre-microsRNA targeting the A1S_3097 gene (signal peptide). Custom TaqMan probe-based real-time qPCRs validated the expression pattern of the selected sRNA candidates shown by RNA-seq. Furthermore, analysis on sRNA ΔA1S_r01, ΔA1S_2505 as well as the over-expressed A1S_3097 mutants revealed no effects on colistin resistance. Our study, for the first time, found the sRNAome signatures of clinical COL with a computational prediction of their targets related to protein synthesis, host-microbe interaction and other different biological functions, including biofilm production, cell-cycle control, virulence, and antibiotic-resistance.
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http://dx.doi.org/10.3389/fmicb.2019.03075DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6978653PMC
January 2020

Results of the Italian infection-Carbapenem Resistance Evaluation Surveillance Trial (iCREST-IT): activity of ceftazidime/avibactam against Enterobacterales isolated from urine.

J Antimicrob Chemother 2020 04;75(4):979-983

Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy.

Objectives: To assess the in vitro antibacterial activity of ceftazidime/avibactam against a recent Italian collection of carbapenem-resistant Enterobacterales (CRE) isolated from urine specimens.

Methods: Consecutive Gram-negative isolates from urine specimens, collected from inpatients in five Italian hospitals during the period October 2016 to February 2017, were screened for CRE phenotype using chromogenic selective medium and identified using MALDI-TOF MS. Antimicrobial susceptibility testing was performed by reference broth microdilution (BMD) and, for ceftazidime/avibactam, also by Etest® CZA. Results were interpreted according to the EUCAST breakpoints. All confirmed CRE were subjected to real-time PCR targeting blaKPC-type, blaVIM-type, blaNDM-type and blaOXA-48-type carbapenemase genes. Non-MBL-producing isolates resistant to ceftazidime/avibactam were subjected to WGS and their resistome and clonality were analysed.

Results: Overall, 318 non-replicate presumptive CRE were collected following screening of 9405 isolates of Enterobacterales (3.4%) on chromogenic selective medium. Molecular analysis revealed that 216 isolates were positive for a carbapenemase gene (of which 92.1%, 2.8%, 1.4% and 1.4% were positive for blaKPC-type, blaOXA-48-type, blaNDM-type and blaVIM-type, respectively). Against the confirmed carbapenemase-producing Enterobacterales (CPE), ceftazidime/avibactam was the most active compound, followed by colistin (susceptibility rates 91.6% and 69.4%, respectively). Compared with BMD, Etest® for ceftazidime/avibactam yielded consistent results (100% category agreement). All class B β-lactamase producers were resistant to ceftazidime/avibactam, while OXA-48 and KPC producers were susceptible, with the exception of seven KPC-producing isolates (4.2%). The latter exhibited an MIC of 16 to >32 mg/L, belonged to ST512, produced KPC-3 and showed alterations in the OmpK35 and Ompk36 porins.

Conclusions: Ceftazidime/avibactam showed potent in vitro activity against a recent Italian collection of CPE from urine.
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http://dx.doi.org/10.1093/jac/dkz547DOI Listing
April 2020

Titration of Igs contained in an intravenous IgM-enriched preparation against selected pathogens involved in sepsis.

Immunobiology 2020 03 14;225(2):151897. Epub 2019 Dec 14.

Laboratory of Molecular Microbiology and Antibiotic Resistance (MMARL), Dept. of Biomedical and Biotechnological Sciences, Via Santa Sofia 97, University of Catania, Catania, Italy. Electronic address:

The goal of our work was to titer the IgG, IgM and IgA in Pentaglobin® (a preparation enriched in IgM), targeting specific surface antigens of Gram-positive and Gram-negative bacteria as well as a C. albicans strain. Lipopolysaccharides from Gram-negative bacteria, peptidoglycan and lipoteichoic acid from the other microorganisms were extracted and used in several ELISA assays in order to determine the titration of immunoglobulins in Pentaglobin® directed towards the aforementioned surface antigens. Our results showed an overall immunoglobulin titer of at least 10 in Pentaglobin® with some exceptions for the IgA titers and for some immunoglobulin titers against E. faecalis, K. pneumoniae and P. aeruginosa. According to these results, Pentaglobin® can be considered as a potential adjuvant for antimicrobial therapy.
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http://dx.doi.org/10.1016/j.imbio.2019.12.006DOI Listing
March 2020

Prevalence of meticillin-resistant and -susceptible coagulase-negative staphylococci with the first detection of the mecC gene among cows, humans and manure in Tunisia.

Int J Antimicrob Agents 2020 Jan 18;55(1):105826. Epub 2019 Oct 18.

University of Manouba, ISBST, BVBGR-LR11ES31, Biotechpole Sidi Thabet, Ariana, Tunisia. Electronic address:

In Europe, a novel mecA homologue - mecC (formerly mecA) - has emerged recently in staphylococci from animals, humans and the environment. This paper reports the first occurrence of the mecC gene in Staphylococcus sciuri from cows and manure in Tunisia and Africa. Forty-nine coagulase-negative staphylococci (CNS) were isolated from the milk of cows with mastitis (n=20), manure (n=20) and human nares (n=9), including 16 Staphylococcus equoruim (32.6%), 12 S. xylosus (24.5%), 12 S. sciuri (24.5%), two S. saprophyticus (4.1%), two S. haemolyticus (4.1%), two S. lentus (4.1%), two S. vitulinus (4.1%) and one S. cohnii (2%). CNS from the three origins carried various resistance genes [mecA, blaZ, tet(K), erm(A), erm(B), msr(A)], suggesting an ongoing genetic exchange among CNS from the three niches. The mecA gene was detected in CNS (n=11) recovered from cows, manure and humans, whereas the mecC gene (n=3) was only detected in CNS from cows and manure. Various staphylococcal cassette chromosome mec (SCCmec) - SCCmec type I (n=1), II (n=3), IV (n=2), V/VII (n=2) and untypeable (n=3) - and diverse pulsed-field gel electrophoresis (PFGE) patterns were observed in mecA-positive CNS. Otherwise, similar SCCmec types and PFGE patterns were found in meticillin-resistant CNS within different farms and origins, showing the potential of SCCmec interspecies exchange and circulation of the same clones of meticillin-resistant CNS in the human-animal-environment interface. This study provides baseline data to support clonal dissemination of CNS between cows, humans and manure, and indicates the possible transmission of the mecC gene to humans in contact with cows and manure.
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http://dx.doi.org/10.1016/j.ijantimicag.2019.10.007DOI Listing
January 2020

Myiasis from Sarcophaga spp in a patient with cutaneous lymphoma.

Infez Med 2019 Sep;27(3):340-344

Department of Biomedical and Biotechnological Sciences, University of Catania, Catania, Italy.

Human autochthonous myiasis is uncommonly reported in Europe. This report describes a case of myiasis of a wound caused by Sarcophaga spp. Suffering from cutaneous lymphoma, the patient showed, at the level of his scalp lesions, the presence of larvae that were removed during curettage surgery; they were subsequently identified as belonging to the genus Sarcophaga. Preservation of these larvae in 10% formalin did not allow identification at the species level using molecular methods.
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September 2019