Genomic and functional analysis of the mucinolytic species Clostridium celatum, Clostridium tertium, and Paraclostridium bifermentans

Mucins are large glycoproteins whose degradation requires the expression of several glycosil hydrolases to catalyze the cleavage of the oligosaccharide chains and release monosaccharides that can be assimilated. In this study, we present a characterization on the strains Clostridium celatum WC0700, Clostridium tertium WC0709, and Paraclostridium bifermentans WC0705. These three strains were previously isolated from enrichment cultures on mucin of fecal samples from healthy subjects and can use mucin as sole carbon and nitrogen source. Genome analysis and in vitro functional analysis of these strains elucidated their physiological and biochemical features. C. celatum WC0700 harbored the highest number of glycosyl hydrolases specific for mucin degradation, while P. bifermentans WC0705 had the least. These predicted differences were confirmed growing the strains on 5 mucin-decorating monosaccharides (L-fucose, N-Acetylneuraminic acid, galactose, N-acetylgalactosamine, and N-acetylglucosamine) as only source of carbon. Fermenting mucin, they all produced formic, acetic, propionic, butyric, isovaleric, and lactic acids, and ethanol; acetic acid was the main primary metabolite. Further catabolic capabilities were investigated, as well as antibiotic susceptibility, biofilm formation, tolerance to oxygen and temperature. The potential pathogenicity of the strains was evaluated through in silico research of virulence factors. The merge between comparative and functional genomics and biochemical/physiological characterization provided a comprehensive view of these mucin degraders, reassuring on the safety of these species and leaving ample scope for deeper investigations on the relationship with the host and for assessing if some relevant health-promoting effect could be ascribed to these SCFA producing species

Antibiotic resistance, virulence factors, phenotyping, and genotyping of non\u2013escherichia coli enterobacterales from the gut microbiota of healthy subjects

Non-Escherichia coli Enterobacterales (NECE) can colonize the human gut and may present virulence determinants and phenotypes that represent severe heath concerns. Most information is available for virulent NECE strains, isolated from patients with an ongoing infection, while the commensal NECE population of healthy subjects is understudied. In this study, 32 NECE strains were isolated from the feces of 20 healthy adults. 16S rRNA gene sequencing and mass spectrometry attributed the isolates to Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Enterobacter kobei, Citrobacter freundii, Citrobacter amalonaticus, Cronobacter sp., and Hafnia alvei, Morganella morganii, and Serratia liquefaciens. Multiplex PCR revealed that K. pneumoniae harbored virulence genes for adhesins (mrkD, ycfM, and kpn) and enterobactin (entB) and, in one case, also for yersiniabactin (ybtS, irp1, irp2, and fyuA). Virulence genes were less numerous in the other NECE species. Biofilm formation was spread across all the species, while curli and cellulose were mainly produced by Citrobacter and Enterobacter. Among the most common antibiotics, amoxicillin-clavulanic acid was the sole against which resistance was observed, only Klebsiella strains being susceptible. The NECE inhabiting the intestine of healthy subjects have traits that may pose a health threat, taking into account the possibility of horizontal gene transfer

Antibiotic resistance, virulence factors, phenotyping, and genotyping of non\u2013escherichia coli enterobacterales from the gut microbiota of healthy subjects

Non-Escherichia coli Enterobacterales (NECE) can colonize the human gut and may present virulence determinants and phenotypes that represent severe heath concerns. Most information is available for virulent NECE strains, isolated from patients with an ongoing infection, while the commensal NECE population of healthy subjects is understudied. In this study, 32 NECE strains were isolated from the feces of 20 healthy adults. 16S rRNA gene sequencing and mass spectrometry attributed the isolates to Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Enterobacter kobei, Citrobacter freundii, Citrobacter amalonaticus, Cronobacter sp., and Hafnia alvei, Morganella morganii, and Serratia liquefaciens. Multiplex PCR revealed that K. pneumoniae harbored virulence genes for adhesins (mrkD, ycfM, and kpn) and enterobactin (entB) and, in one case, also for yersiniabactin (ybtS, irp1, irp2, and fyuA). Virulence genes were less numerous in the other NECE species. Biofilm formation was spread across all the species, while curli and cellulose were mainly produced by Citrobacter and Enterobacter. Among the most common antibiotics, amoxicillin-clavulanic acid was the sole against which resistance was observed, only Klebsiella strains being susceptible. The NECE inhabiting the intestine of healthy subjects have traits that may pose a health threat, taking into account the possibility of horizontal gene transfer

Antibiotic Resistance, Virulence Factors, Phenotyping, and Genotyping of E. coli Isolated from the Feces of Healthy Subjects

Escherichia coli may innocuously colonize the intestine of healthy subjects or may instigate infections in the gut or in other districts. This study investigated intestinal E. coli isolated from 20 healthy adults. Fifty-one strains were genotyped by molecular fingerprinting and analyzed for genetic and phenotypic traits, encompassing the profile of antibiotic resistance, biofilm production, the presence of surface structures (such as curli and cellulose), and their performance as recipients in conjugation experiments. A phylogroup classification and analysis of 34 virulence determinants, together with genes associated to the pks island (polyketide-peptide genotoxin colibactin) and conjugative elements, was performed. Most of the strains belonged to the phylogroups B1 and B2. The different phylogroups were separated in a principal coordinate space, considering both genetic and functional features, but not considering pulsed-field gel electrophoresis. Within the B2 and F strains, 12 shared the pattern of virulence genes with potential uropathogens. Forty-nine strains were sensitive to all the tested antibiotics. Strains similar to the potential pathogens innocuously inhabited the gut of healthy subjects. However, they may potentially act as etiologic agents of extra-intestinal infections and are susceptible to a wide range of antibiotics. Nevertheless, there is still the possibility to control infections with antibiotic therapy

Black soldier fly (Hermetia illucens L.) as a high-potential agent for bioconversion of municipal primary sewage sludge

The treatment of municipal wastewater produces clean water and sewage sludge (MSS), the management of which has become a serious problem in Europe. The typical destination of MSS is to spread it on land, but the presence of heavy metals and pollutants raises environmental and health concerns. Bioconversion mediated by larvae of black soldier fly (BSFL) Hermetia illucens (Diptera, Stratiomyidae: Hermetiinae) may be a strategy for managing MSS. The process adds value by generating larvae which contain proteins and lipids that are suitable for feed and/or for industrial or energy applications, and a residue as soil conditioner. MSS from the treatment plant of Ladispoli (Rome province) was mixed with an artificial fly diet at 50% and 75% (fresh weight basis) to feed BSFL. Larval performance, substrate reduction, and the concentrations of 12 metals in the initial and residual substrates and in larval bodies at the end of the experiments were assessed. Larval survival (> 96%) was not affected. Larval weight, larval development, larval protein and lipid content, and waste reduction increased in proportion the increase of the co-substrate (fly diet). The concentration of most of the 12 elements in the residue was reduced and, in the cases of Cu and Zn, the quantities dropped under the Italian national maximum permissible content for fertilizers. The content of metals in mature larvae did not exceed the maximum allowed concentration in raw material for feed for the European Directive. This study contributes to highlight the potential of BSF for MSS recovery and its valorization. The proportion of fly diet in the mixture influenced the process, and the one with the highest co-substrate percentage performed best. Future research using other wastes or by-products as co-substrate of MSS should be explored to determine their suitability

The LuGRE project: a scientific opportunity to study GNSS signals at the Moon

The Lunar GNSS Receiver Experiment (LuGRE) is a joint NASA-Italian Space Agency (ASI) payload on the Firefly Blue Ghost Mission 1 with the goal to demonstrate GNSS-based positioning, navigation, and timing at the Moon. When launched, LuGRE will collect GPS and Galileo measurements in transit between Earth and the Moon, in lunar orbit, and on the lunar surface, and will conduct onboard and ground-based navigation experiments using the collected data. These investigations will be based on the observation of the data collected by a custom development performed by the company Qascom, based on the Qascom QN400-Space GNSS receiver. The receiver is able to provide, PVT solutions, the GNSS raw observables obtained by the real time operation, as well as snapshots of IF digital samples collected by the RF front-end at frequencies L1/E1 and L5/E5. These data will be the input for the different science investigations, that require then the development of proper analysis tools that will be the core of the ground segment during the mission. The current work done by the science team of NASA and ASI, which is supported by a research team at Politecnico di Torino, is planning the data acquisitions during the time windows dedicated to the LuGRE payload in the checkout, transit and surface mission phases

Effetto dell'estratto di Asphodelus ramosus (L), specie mediterranea, sul controllo di patogeni ed insetti di interesse agrario

L'Asfodelo è una pianta erbacea originaria del sud Europa, Africa, Medio Oriente e sub continente Indiano, ma esprime la sua maggiore diversità soprattutto nel bacino del Mediterraneo. Fin dall’antichità sono riconosciute le sue proprietà terapeutiche ed alimentari essendo ricca in flavonoidi, come luteolina, isovitexina, quercetina, e acidi fenolici (e.g. acido caffeico, gallico e clorogenico) acidi grassi e polisaccaridi (“inulina-type fruttoligosaccarides”). Il documento del Green Deal Europeo, ‘Farm to Fork’, ha posto l’obbiettivo, entro il 2030, di una riduzione del 50% dell'uso di sostanze chimiche fitosanitarie. In questo contesto l’uso dei fito-estratti naturali rappresenta una possibile alternativa all’uso dei composti chimici di sintesi da integrare ad altre soluzioni per una gestione sostenibile del sistema produttivo. A questo scopo si è voluto determinare l’effetto di estratti acquosi da radice di A. ramosus (L) a diverse concentrazioni sia sullo sviluppo in vitro di Botrytis cinerea, patogeno ubiquitario in grado di infettare molte specie vegetali, che sulla schiusa di uova, e sugli stadi giovanili della cimice asiatica, Halimorpha halys (Stål), specie invasiva polifaga su cui non sono ancora disponibili efficaci sistemi di controllo. I risultati ottenuti evidenziano un effetto inibitorio dell’estratto di asfodelo, in funzione della sua concentrazione ed un efficace effetto insetticida sulla cimice asiatica. L’estratto è stato analizzato anche per le sue caratteristiche fisico/chimiche.The Asphodelus genus, is an herbaceous plant originating in southern Europe, Africa, the Middle East and the Indian subcontinent, which exhibits a great genetic variability in the Mediterranean area. Its therapeutic properties have been recognised since the ancient times, as it is rich in flavonoids, such as luteolin, isovitexin, quercetin, and phenolic acids (e.g. caffeic, gallic and chlorogenic acids) fatty acids and polysaccharides ('inulin-type fructooligosaccharides'). The European Green Deal, in particular the 'Farm to Fork', established the objective of 50% reduction of pesticides use by 2030. In that context, the use of natural phytoextracts represents a possible alternative to the application of synthetic chemical compounds, coupled with other solutions for a sustainable crop management system. The aim of this study was to determine the effect of aqueous extracts of Asphodelus ramosus roots at different concentrations on both, in vitro development of Botrytis cinerea, a ubiquitous pathogen able to infect many crops, and on eggs and juvenile stages of the Asian bedbug, Halimorpha halys (Stål), an invasive polyphagous insect, for which no efficient control systems are yet available. The results obtained show an inhibitory effect of the A. ramosus extract, according to the concentrations, and a significant insecticidal effect on the Asian bedbug. The extract was also analysed for its physic/chemical characteristics

[SENTIERI Project: rationale and objectives].

The National strategic programme “Environment and health” was funded by the Ministry of Health and coordinated by the Istituto Superiore di Sanità. The Programme focused on “the health impact associated with residence in polluted sites, in areas affected by waste disposal/incineration facilities and exposure to air pollution in urban areas” and included six research projects (with a total of 41 units). One of the six projects called “Risk to health in polluted sites: exposure assessment, biomonitoring and epidemiological characterization” comprised ten units, eight of which were devoted to SENTIERI Project (Mortality study of residents in italian polluted sites). The Project started in 2007 and was completed in December 2010. The results are published in two supplements of Epidemiologia & Prevenzione. SENTIERI Project is the first report systematically discussing cause-specific mortality in populations living in IPSs. The results are based on the a priori evaluation of the evidence presented in the previous supplement (Pirastu et al., 2010), and on the consequent etiological hypotheses; the discussion takes into account possible confounding from socioeconomic deprivation. In this second Supplement, which is divided into two sections, the results of the mortality analysis for the 44 IPS included in the Project for the years 1995-2002 are presented. Some comments and operational guidance on further epidemiological characterization of these areas are also provided. The second section of the present Supplement is devoted to a thorough analysis of SENTIERI Project and to its future development. It includes several chapters describing new activities that are under way or planned. It is the Working Group’s opinion, and in particular of the editors, that additional epidemiological data about populations residing in IPSs are necessary for a deeper understanding of the health impact of polluted sites, and an appropriate detection of priority intervention in environmental remediation