Salvatore Oliveri

Environmental distribution of Cryptococcus neoformans and C. gattii around the Mediterranean basin

In order to elucidate the distribution of Cryptococcus neoformans and C. gattii in the Mediterranean basin, an extensive environmental survey was carried out during 2012-2015. A total of 302 sites located in 12 countries were sampled, 6436 samples from 3765 trees were collected and 5% of trees were found to be colonized by cryptococcal yeasts. Cryptococcus neoformans was isolated from 177 trees and C. gattii from 13. Cryptococcus neoformans colonized 27% of Ceratonia, 10% of Olea, Platanus and Prunus trees and a lower percentage of other tree genera. The 13 C. gattii isolates were collected from five Eucalyptus, four Ceratonia, two Pinus and two Olea trees. Cryptococcus neoformans was distributed all around the Mediterranean basin, whereas C. gattii was isolated in Greece, Southern Italy and Spain, in agreement with previous findings from both clinical and environmental sources. Among C. neoformans isolates, VNI was the prevalent molecular type but VNII, VNIV and VNIII hybrid strains were also isolated. With the exception of a single VGIV isolate, all C. gattii isolates were VGI. The results confirmed the presence of both Cryptococcus species in the Mediterranean environment, and showed that both carob and olive trees represent an important niche for these yeasts.

Experience with the Platelia Candida ELISA for the diagnosis of invasive candidosis in neonatal patients

This preliminary study evaluated the use of the Platelia Candida antigen kit for the diagnosis of invasive candidosis in 70 of 184 pre-term infants admitted to a neonatal intensive care unit between March 2004 and March 2006. The frequency of confirmed candidaemia was 6.5%. The sensitivity and specificity of the assay were 94.4% and 94.2%, respectively, with a positive predictive value of 85% and a negative predictive value of 98%. These results suggest that the inclusion of regular serological surveillance for mannanaemia in some pre-term infants would complement blood cultures for the early detection of candidosis.

Malassezia furfur fungaemia in a neonatal patient detected by lysis‐centrifugation blood culture method: first case reported in Italy

Malassezia yeast typically colonise the skin of healthy individuals (Morishita N, Sei Y. Mycopathologia 2006; 162: 373–6). However, under some conditions, they are associated with a number of skin infections in adults (Tarazooie B et al., BMC Dermatol 2004; 4: 5). Also newborns can be colonised. In a study of hospitalised neonates, Malassezia spp. were isolated from 68.7%, with Malassezia furfur the most isolated species. The high prevalence of colonisation encountered in hospitalised neonates highlights the potential danger of nosocomial infections caused by Malassezia sp., particularly in those instances in which intravenous lipids or indwelling catheters are needed (Gupta AK et al., J Am Acad Dermatol 2004; 51: 785–98). These yeast are important agents of catheter-associated sepsis, primarily in low-birth-weight infants receiving lipids through a central venous line (Zomorodain K et al., Pediatric Dermatology 2008; 3: 312–6). Invasive infections caused by species of Candida or Malassezia have been documented in the neonatal intensive care units (NICU) and are often associated with significant morbidity and mortality (Devlin RK, Adv Neonatal Care 2006; 6: 68– 77). Malassezia sp. remains probably underdiagnosed, as it does not routinely grow in the usual cultures and requires lipid-supplemented media for in vitro growth. The Isolator system has been reported to provide greater sensitivity than conventional broth blood culture systems in recovering fungal organisms from the blood (Berenguer J et al., Diagn Microbiol Infect Dis 1993; 17: 103–9). In the literature, cases of Malassezia fungaemia in Italy have not been reported. Case report

Comparison between Disk Diffusion and Microdilution Methods for Determining Susceptibility of Clinical Fungal Isolates to Caspofungin

ABSTRACT We compared the caspofungin (CAS) susceptibility testing results generated by the disk diffusion (DD) assay with the results of the Clinical and Laboratory Standards Institute (CLSI) broth microdilution (BD) reference method for 106 yeast isolates. The isolates represented 11 different fungal species, including Candida albicans (n = 50), C. parapsilosis (n = 10), C. glabrata (n = 10), C. tropicalis (n = 10), C. guillermondii (n = 6), C. rugosa (n = 5), C. krusei (n = 5), C. kefyr (n = 2), C. pelliculosa (n = 2), Saccharomyces cerevisiae (n = 3), and Geotrichum candidum (n = 3). The DD assay was performed in supplemented Mueller-Hinton agar with CAS, which was tested at concentrations of 2, 10, and 25 μg per disk. MICs and inhibition zone diameters were evaluated at 24 and 48 h. In general, the results obtained by the DD assay correlated well with those obtained by the BD method. In particular, a significant correlation between methods was observed when CAS was used at concentration of 2 μg/disk at a reading time of either 24 or 48 h.

Detection of fungal DNA in lysis–centrifugation blood culture for the diagnosis of invasive candidiasis in neonatal patients

We report data concerning the detection of fungal DNA directly from lysis-centrifugation blood culture to assess its value in the detection of fungaemia in 86 of the 347 patients admitted to the neonatal intensive-care unit between January 2009 and December 2010. The sensitivity and specificity of the PCR were 87.5% and 98.5%, respectively, with a positive predictive value of 93.3% and a negative predictive value of 97.1%. Detection of fungal DNA directly from blood culture Isolator 1.5 microbial tubes, without prior cultivation, is a promising approach for the rapid detection of Candida spp. in neonates with suspected candidaemia.

Evaluation of the disk diffusion method compared to the microdilution method in susceptibility testing of anidulafungin against filamentous fungi.

ABSTRACT Susceptibility testing of anidulafungin (AFG) against 32 mold isolates showed an excellent correlation between disk diffusion (DD) and broth microdilution methods. Based on our data, a 2-μg disk of AFG and a 24-h reading time might represent the best parameters for AFG DD testing against filamentous fungi.

Azole-Resistance in Aspergillus terreus and Related Species: An Emerging Problem or a Rare Phenomenon?

Objectives: Invasive mold infections associated with Aspergillus species are a significant cause of mortality in immunocompromised patients. The most frequently occurring aetiological pathogens are members of the Aspergillus section Fumigati followed by members of the section Terrei. The frequency of Aspergillus terreus and related (cryptic) species in clinical specimens, as well as the percentage of azole-resistant strains remains to be studied. Methods: A global set (n = 498) of A. terreus and phenotypically related isolates was molecularly identified (beta-tubulin), tested for antifungal susceptibility against posaconazole, voriconazole, and itraconazole, and resistant phenotypes were correlated with point mutations in the cyp51A gene. Results: The majority of isolates was identified as A. terreus (86.8%), followed by A. citrinoterreus (8.4%), A. hortai (2.6%), A. alabamensis (1.6%), A. neoafricanus (0.2%), and A. floccosus (0.2%). One isolate failed to match a known Aspergillus sp., but was found most closely related to A. alabamensis. According to EUCAST clinical breakpoints azole resistance was detected in 5.4% of all tested isolates, 6.2% of A. terreus sensu stricto (s.s.) were posaconazole-resistant. Posaconazole resistance differed geographically and ranged from 0% in the Czech Republic, Greece, and Turkey to 13.7% in Germany. In contrast, azole resistance among cryptic species was rare 2 out of 66 isolates and was observed only in one A. citrinoterreus and one A. alabamensis isolate. The most affected amino acid position of the Cyp51A gene correlating with the posaconazole resistant phenotype was M217, which was found in the variation M217T and M217V. Conclusions: Aspergillus terreus was most prevalent, followed by A. citrinoterreus. Posaconazole was the most potent drug against A. terreus, but 5.4% of A. terreus sensu stricto showed resistance against this azole. In Austria, Germany, and the United Kingdom posaconazole-resistance in all A. terreus isolates was higher than 10%, resistance against voriconazole was rare and absent for itraconazole.

Error control in UAV image acquisitions for 3D reconstruction of extensive architectures

This work describes a simple, fast, and robust method for identifying, checking and managing the overlapping image keypoints for 3D reconstruction of large objects with numerous geometric singularities and multiple features at different lighting levels. In particular a precision 3D reconstruction of an extensive architecture captured by aerial digital photogrammetry using Unmanned Aerial Vehicles (UAV) is developed. The method was experimentally applied to survey and reconstruct the ‘Saraceni’ Bridge’ at Adrano (Sicily), a valuable example of Roman architecture in brick of historical/cultural interest. The variety of features and different lighting levels required robust self-correlation techniques which would recognise features sometimes even smaller than a pixel in the digital images so as to automatically identify the keypoints necessary for image overlapping and 3D reconstruction. Feature Based Matching (FBM) was used for the low lighting areas like the intrados and the inner arch surfaces, and Area Based Matching (ABM) was used in conjunction to capture the sides and upper surfaces of the bridge. Applying SIFT (Scale Invariant Feature Transform) algorithm during capture helped find distinct features invariant to position, scale and rotation as well as robust for the affinity transformations (changes in scale, rotation, size and position) and lighting variations which are particularly effective in image overlapping. Errors were compared with surveys by total station theodolites, GPS and laser systems. The method can facilitate reconstruction of the most difficult to access parts like the arch intrados and the bridge cavities with high correlation indices.

Dynamics of a high-performance motorcycle by an advanced multibody/control co-simulation

The present work aims at the development of an advanced control system implemented through Adams/View-Matlab/Simulink co-simulation for a high-performance motorcycle dynamics study. In particular, the purpose of this study is to create a model able to consider several aspects of the rider–motorbike dynamic simulation and its control system, generally treated separately in the literature, making also use of an original and accurate modelling of the rider. From a previous multi-body model of motorcycle/virtual rider, developed by the authors, a flexible tool is created to simulate system dynamics to follow any trajectory at a prescribed velocity profile. Considering high-performance motorcycle dynamics are greatly influenced by the rider’s weight, his movements have been accurately replicated to obtain the most realistic results. To simulate the passive impedance of rider’s arms, a torque was applied to the steering as per the literature. The aerodynamic force was modelled as a function of kinematics variables and rider’s posture. The control system is very flexible and adaptable to different manoeuvres realistically reproducing engine and braking performance, steering torque and rider movements. Numerical results show that the control system can accurately direct the motorcycle/rider system along an entire lap of the Monza circuit, following a desired path at a given velocity profile. The model developed allows a complete view of the motorbike-rider dynamic behaviour thus being useful during both design phase and set-up, with a considerable saving in terms of both cost and time; it can also evaluate the influence on the system dynamics of riders with different anatomical characteristics and driving styles.

An advanced multibody model for evaluating rider’s influence on motorcycle dynamics

The dynamics of a high-performance motorcycle are greatly influenced by the rider’s weight and movements especially when the power-to-weight ratio is very high. Generally in motor vehicles, the driver’s/rider’s weight is a significant fraction of the entire system. This work is about ADAMS/View multibody modelling of a motorcycle and virtual rider who simulates handlebar interaction and saddle sliding. In the literature, the rider’s influence is unrealistic being limited to considering him as a concentrated mass or in other cases as a fixed passive system. Even vehicle modelling is often inaccurate, referring at best to simplified models of rigid bodies. In this work, the vehicle and rider have been accurately modelled to most realistically reproduce the dynamic behaviour of the system. The motorcycle was modelled with 12 bodies incorporating concentrated flexibility for the two suspension units and considering the chassis as a flexible body using modal synthesis. The virtual rider is made up of 15 rigid bodies and has 28 degrees of freedom. To study the effects on the motorcycle of the rider’s movements as well as the motorcycle’s dynamics and performance, a monitoring system similar to that in the literature was used to read handlebar torque and engine and braking torque. Furthermore, in the literature there are simulations of standard manoeuvres whereas in this work an entire lap of Monza was simulated. There were simulations of a fixed and mobile rider validating the model in advance and thereafter monitoring the most significant dynamic parameters. The multibody model provides useful results at the design phase and insights into the whole vehicle/rider dynamic to setup all the reference parameters for immediately evaluating system effects.