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Il progetto “Impact of Nanoparticles in Environmental Sustainability and Ecotoxicity” (INESE), finanziato dall’Istituto Italiano di Tecnologia (IIT), ha studiato gli effetti delle nanoparticelle (NPs) ingegnerizzate su diversi ecosistemi (acquatici e terrestri), proponendo differenti sistemi biologici di studio (piante acquatiche, ricci di mare, organismi del suolo, pomodoro-impollinatore, ecc). In particolare l’UO dell’Università di Bologna ha focalizzato l’attenzione sullo studio dei modelli di interazione tra particelle di suolo, microorganismi (biomassa microbica), invertebrati (Lumbricus rubellus) e pianta (Solanum lycopersicum). In questa sede verranno riportati i risultati ottenuti dall’impatto delle nanoparticelle di argento (Ag-NPs) nell’ecosistema suolo. La crescita esponenziale dell’uso di materiali contenenti Ag-NPs è dovuta alla loro importante azione antimicrobica, sempre più sfruttata in diversi campi: dalla medicina (bendaggi) all’abbigliamento (magliette, calzini etc.), ai prodotti di igiene personale (spazzolini da denti), dalla sanificazione delle acque reflue con conseguente produzione di biosolidi impiegati come ammendanti agricoli, all’utilizzo in agricoltura di prodotti dedicati ("Nano-Argentum 10", Fa Nanosys, Svizzera). Studi basati su monitoraggio registrano l’aumento di Ag-NPs nei diversi comparti ambientali (aria-acqua e suolo) e dimostrano che il suolo è destinato ad esserne il maggior ricettore. Il C contenuto nella biomassa microbica del suolo (Cmic) è significativamente diminuita dopo l'esposizione ad Ag-NPs rispetto al controllo, e ciò viene anche confermato dalla diminuzione del DNA microbico estraibile e dall’analisi PCR-DGGE. Quest’ultima ha mostrato la capacità battericida dell’Ag-NPs, riducendo la comunità microbica di un suolo forestale a solo 5 ceppi di batteri resistenti. Riguardo all’impatto sugli invertebrati, il maggior contenuto di Ag rispetto al controllo trovato nei tessuti dei lombrichi è stato riconfermato dall’analisi ESEM che ha mostrato la presenza di Ag-NPs nell’intestino dei lombrichi, dopo depurazione. Inoltre l’esposizione di questi organismi ad Ag-NPs ha causato significativi cambiamenti nella composizione lipidica dei tessuti, rilevando quindi condizioni di stress. La somministrazione di Ag-NPs alle piante di pomodoro tramite l’acqua di irrigazione ha messo in evidenza come le NPs possano essere traslocate dalle radici alle foglie e infine al frutto edule. I potenziali rischi per la salute umana ed ambientale dovuti all’incontrollata diffusione delle NPs costituiscono motivi di grave preoccupazione per la comunità scientifica internazionale.

Presentazione La Sezione di Studio di Radiologia Informatica della SIRM con questo documento ha voluto condurre un'approfondita . analisi delle problematiche relative all'informatizzazione del Servizio di Radiologia con l'intento di contribuire alla definizione dei requisiti dei Sistemi Informativi di Radiologia (RIS). E' intuitivo che lo strumento informatico, unito a buone capacità organizzative e gestionali, è un naturale alleato del Radiologo nel processo di continuo miglioramento della qualità del servizio offerto. D'altra parte lo strumento informatico non è unico né perfetto. Pur parlando solo del RIS, noi sappiamo come sul mercato esistano molti prodotti, tutti di pregio eppure a volte molto diversi tra di loro per caratteristiche tecniche, usabilità e funzioni disponibili. La stessa legge di mercato che impone ai prodotti di differenziarsi può rendere difficile coglierne i pregi e i difetti e a volte capita che proprio quella funzionalità a cui non si era pensato in sede di valutazione preliminare del prodotto si riveli poi critica nella realtà operativa. In questo documento si tenta di definire i requisiti che un Sistema Informativo di Radiologia dovrebbe soddisfare per essere considerato allo stato dell'arte, dove con questo temine si intende non tanto lo stato dell'arte tecnologico ma la capacità di migliorare "al massimo grado" la qualità del lavoro del Radiologo e dei suoi Collaboratori. E' difficile pensare a questo documento come definitivo. Esso dovrà piuttosto servire come base per una più ampia discussione che coinvolga Radiologi, Informatici, Tecnici e le stesse Ditte produttrici. In futuro piacerebbe poter disporre di vere e proprie Linee Guida che facilitino il dialogo tra il Radiologo e l'informatico, nella ricerca di un linguaggio comune e nel rispetto delle reciproche esigenze e competenze.

By using abdominal ultrasonography (UlS), deep nodes were detected in 41 of 85 (48%) HIV-1 positive subjects, most of them heroin addicts, but in none of 85 healthy HIV-negative controls. Computerized tomography, performed in 10 cases of lymphadenopathy, invariably confirmed the UlS findings. Prevalence [asymptomatic carriers: 8/15 (53%); PGL patients: 8/18 (44%); ARC: 13/27 (48%); AIDS: 12/25 (48%)], number, size, and site of deep nodes were comparable among the different CDC groups. No correlation was found between abdominal and superficial lymphadenopathy. Median serum concentrations of gammaglobulins (g/dl) and IgG (mg/dl) were higher in patients with than without deep nodes (2.25 vs 1.87 and 2540 vs 1900, respectively) (p0.01) as well as in cases with than without superficial nodes (2.15 vs 1.80 and 2340 vs 1941, respectively) (p0.05). Abdominal lymphadenopathy occurred during all stages of HIV infection even in asymptomatic carriers: this should be considered in the differential diagnosis of UlS-detected deep nodes. Enlargement of either deep or superficial nodes seems to reflect a state of polyclonal B cell activation.

Serum samples from 66 seropositive subjects (56 with a history of intravenous drug abuse), including asymptomatic carriers and patients with persistent generalised lymphadenopathy (PGL), AIDS related complex (ARC), and AIDS, were tested by indirect immunofluorescence on rat tissue sections and HEp-2 cells for the presence of antibodies to nuclei, smooth muscle, intermediate filaments (anti-IMF) and microfilaments (anti-MF). Counterimmunoelectrophoresis was also used to detect antibodies to extractable nuclear antigens. Smooth muscle antibodies with the V pattern or antinuclear antibodies, mainly of the speckled type, or anti-IMF, occurred in 35 cases, being widely distributed in all groups. Such an autoantibody response resembles the "viral" autoimmunity described in various infectious diseases and in particular that of non-A, non-B post-transfusion hepatitis. Autoantibodies may be of some prognostic relevance, as the prevalence of smooth muscle antibodies V increased as the disease progressed (asymptomatic carriers 20%, those with PGL 29%, those with ARC 47%, and those with AIDS 63%. In the PGL group autoantibody positivity correlated with the presence of skin anergy. The fact that autoantibodies were more frequently detected in patients with circulating immune complexes suggests that these can contain autoantibodies and the corresponding autoantigens.

Using counterimmunoelectrophoresis (CIE), serum antibodies to rabbit thymus extractable antigens were detected in 15% (38/259) of patients with chronic liver disease (CLD) of various aetiologies and 33% (41/124) of patients with miscellaneous connective tissue diseases (CTD). A remarkable diversity of precipitating systems was apparent among cases with the two classes of disorders. All the five systems found in CLD (XR, XR2, SS-B, XR3, XR4) were associated mostly with immunological hepatic disorders. In the 52 autoimmune hepatitis cases, XR was mainly detected (29%), whereas in the 82 primary biliary cirrhosis patients the whole spectrum of reactivities was represented (XR: 11%, XR2: 10%, SS-B and XR3: 2% each, XR4: 1%). XR proved to be closely associated with smooth muscle antibodies (SMA, detected by indirect immunofluorescence on rat kidney sections) both qualitatively and quantitatively. Since all SMA positive sera with anti-actin specificity (SMAT, SMAG) were XR positive and purified actin could absorb out XR CIE reactivity, the hypothesis is made that a cross-reaction occurs between XR antigen and actin epitope(s).

Antibodies against thymus epithelial cells (anti-TEC) and the basal cell layer (BCLA) of squamous epithelia have been described in association with HDV-related chronic liver disease (CLD). Data are lacking on their presence during nAnB virus infection. Sera from 51 patients with nAnB post-transfusion hepatitis, including acute and chronic cases diagnosed during a prospective study on candidates for cardiac surgery, and 167 with various forms of CLD were tested for the presence of anti-TEC and BCLA using indirect immunofluorescence on human thymus and rat forestomach sections. Both antibodies mainly occurred in nAnB, HDV and cryptogenic CLD (anti-TEC: 51%, 47% and 42%; BCLA: 29%, 38% and 31%, respectively). The prevalence of anti-TEC in nAnB CLD turned out to be higher than that recorded in alcoholic, HBV-related, autoimmune, liver and kidney microsomal antibody positive CLD and primary biliary cirrhosis (p ranging from less than 0.03 to less than 0.0004). Two monoclonal antibodies (Mabs) to cytokeratins gave a pattern superimposable on that of spontaneous anti-TEC (both Mabs) and BCLA (only one). Antibodies against epithelial constituents, presumably targeting cytokeratin-associated antigens, occur not only in HDV CLD, as previously reported, but also in nAnB CLD, where they might represent a diagnostic aid, due to the unavailability of reliable serological markers of nAnB infection. The close similarity of anti-TEC and BCLA status between nAnB and cryptogenic CLD suggests a nAnB etiology of at least a proportion of chronic liver patients at present scored as cryptogenic.

BACKGROUND: Coronavirus disease 2019 (COVID-19) has spread worldwide determining dramatic impacts on healthcare systems. Early identification of high-risk parameters is required in order to provide the best therapeutic approach. Coronary, thoracic aorta and aortic valve calcium can be measured from a non-gated chest computer tomography (CT) and are validated predictors of cardiovascular events and all-cause mortality. However, their prognostic role in acute systemic inflammatory diseases, such as COVID-19, has not been investigated. OBJECTIVES: The aim was to evaluate the association of coronary artery calcium and total thoracic calcium on in-hospital mortality in COVID-19 patients. METHODS: 1093 consecutive patients from 16 Italian hospitals with a positive swab for COVID-19 and an admission chest CT for pneumonia severity assessment were included. At CT, coronary, aortic valve and thoracic aorta calcium were qualitatively and quantitatively evaluated separately and combined together (total thoracic calcium) by a central Core-lab blinded to patients' outcomes. RESULTS: Non-survivors compared to survivors had higher coronary artery [Agatston (467.76 ​± ​570.92 vs 206.80 ​± ​424.13 ​mm2, p ​

Background A connection between amyotrophic lateral sclerosis (ALS) and altered gut microbiota composition has previously been reported in animal models. This work is the first prospective longitudinal study addressing the microbiota composition in ALS patients and the impact of a probiotic supplementation on the gut microbiota and disease progression. Methods Fifty patients and 50 matched controls were enrolled. The microbial profile of stool samples from patients and controls was analyzed via PCR-Denaturing Gradient Gel Electrophoresis, and the main microbial groups quantified via qPCR. The whole microbiota was then analyzed via next generation sequencing after amplification of the V3–V4 region of 16S rDNA. Patients were then randomized to receive probiotic treatment or placebo and followed up for 6 months with ALSFRS-R, BMI, and FVC%. Results The results demonstrate that the gut microbiota of ALS patients is characterized by some differences with respect to controls, regardless of the disability degree. Moreover, the gut microbiota composition changes during the course of the disease as demonstrated by the significant decrease in the number of observed operational taxonomic unit during the follow-up. Interestingly, an unbalance between potentially protective microbial groups, such as Bacteroidetes, and other with potential neurotoxic or pro-inflammatory activity, such as Cyanobacteria, has been shown. The 6-month probiotic treatment influenced the gut microbial composition; however, it did not bring the biodiversity of intestinal microbiota of patients closer to that of control subjects and no influence on the progression of the disease measured by ALSFRS-R was demonstrated. Conclusions Our study poses the bases for larger clinical studies to characterize the microbiota changes as a novel ALS biomarker and to test new microbial strategy to ameliorate the health status of the gut. Trial registration CE 107/14, approved by the Ethics Committee of the “Maggiore della Carità” University Hospital, Italy.

Background and aims: As potent allelochemicals, flavonoids are believed to be associated with the development of soil-borne diseases. Fusarium species infection is responsible for soybean [Glycine max(L.) Merr.] root rot. However, it is uncertain if and how flavonoids influence rhizosphere Fusarium communities under consecutive soybean monoculture. Methods: Quantitative real-time PCR and pyrosequencing were used to study the Fusarium community. Spectrophotometric techniques and ultra-performance liquid chromatography (UPLC) were used to quantify flavonoids and daidzein and genistein levels, respectively. Results: In field soil, the sizes of Fusarium community were higher from 6 to 13years monoculture than from 1 to 3years. The abundance of Fusarium community was significantly positively correlated with rhizosphere concentrations of the flavonoids and isoflavonoids daidzein and genistein levels. Consecutive monocultured soil selectively inhibited or stimulated certain Fusarium species, F. oxysporum remaining the most abundant. The application of daidzein and a mixture of daidzein and genistein to soil affected the Fusarium community depending on the incubation time and ranging from inhibition to promotion over time. Conclusions: Consecutive soybean monoculture results in shifts in the composition and size of rhizosphere Fusarium community. The Fusarium community was strongly influenced by total flavonoids, in particular daidzein and genistein, rather than soil properties.

Several studies have outlined that changes in the honeybee gut microbial composition may impair important metabolic functions supporting the honeybees’ life. Gut dysbiosis may be caused by diseases like Nosema ceranae or by other anthropic, environmental or experimental stressors. The present work contributes to increasing knowledge on the dynamics of the gut microbiome acquisition in caged honeybees, an experimental condition frequently adopted by researchers, with or without infection with N. ceranae, and fed with a bacterial mixture to control N. ceranae development. Changes of the gut microbiota were elucidated comparing microbial profile of caged and open-field reared honeybees. The absolute abundance of the major gut microbial taxa was studied with both NGS and qPCR approaches, whereas changes in the functionality were based on RAST annotations and manually curated. In general, all caged honeybees showed important changes in the gut microbiota, with $$\gamma$$ γ -proteobacteria (Frischella, Gilliamella and Snodgrassella) lacking in all caged experimental groups. Caged honeybees infected with N. ceranae showed also a strong colonization of environmental taxa like Citrobacter, Cosenzaea and Morganella, as well as possibly pathogenic bacteria such as Serratia. The colonization of Serratia did not occur in presence of the bacterial mixture. The functionality prediction revealed that environmental bacteria or the supplemented bacterial mixture increased the metabolic potential of the honeybee gut microbiome compared to field and caged controls.

Intensive conventional farm management, characterized by high agrochemicals input, could alter the composition of microbial communities with potential negative effects on both functional traits and the ecosystem services provided. In this study, we investigated the gut microbial composition of a high ecological relevance carabid Pseudoophonus rufipes, sampled in two fields subjected to conventional and organic management practices. Carabids’ gut microbiota was analyzed via qPCR and NGS. Profound differences between the microbial composition of organic and conventional samples were detected: the abundance of Tenericutes and Proteobacteria was significant higher in organic and conventional samples, respectively. Spiroplasmataceae and Bifidobacteriaceae families were significantly more abundant in samples from organic management, while Enterococcaceae, Morganellaceae and Yersiniaceae were more abundant in samples from conventional management. The diverse gut microbial composition of insects between the two management systems is related to the pressure of environmental stressors and it may representing an important bioindication of ecological functions and services provided by a carabid species.

The role of a balanced gut microbiota to maintain health and prevent diseases is largely established in humans and livestock. Conversely, in honeybees, studies on gut microbiota perturbations by external factors have started only recently. Natural methods alternative to chemical products to preserve honeybee health have been proposed, but their effect on the gut microbiota has not been examined in detail. This study aims to investigate the effect of the administration of a bacterial mixture of bifidobacteria and Lactobacillaceae and a commercial product HiveAliveTM on honeybee gut microbiota. The study was developed in 18 hives of about 2500 bees, with six replicates for each experimental condition for a total of three experimental groups. The absolute abundance of main microbial taxa was studied using qPCR and NGS. The results showed that the majority of the administered strains were detected in the gut. On the whole, great perturbations upon the administration of the bacterial mixture and the plant-based commercial product were not observed in the gut microbiota. Significant variations with respect to the untreated control were only observed for Snodgrassella sp. for the bacterial mixture, Bartonella sp. in HiveAliveTM and Bombilactobacillus sp. for both. Therefore, the studied approaches are respectful of the honeybee microbiota composition, conceivably without compromising the bee nutritional, social and ecological functions.

Several studies have outlined that a balanced gut microbiota offers metabolic and protective functions supporting honeybee health and performance. The present work contributes to increasing knowledge on the impact on the honeybee gut microbiota of the three most common veterinary drugs (oxytetracycline, sulfonamides, and tylosin). The study was designed with a semi-field approach in micro-hives containing about 500 honeybees. Micro-hives were located in an incubator during the day and moved outdoors in the late afternoon, considering the restrictions on the use of antibiotics in the open field but allowing a certain freedom to honeybees; 6 replicates were considered for each treatment. The absolute abundance of the major gut microbial taxa in newly eclosed individuals was studied with qPCR and next-generation sequencing. Antimicrobial resistance genes for the target antibiotics were also monitored using a qPCR approach. The results showed that the total amount of gut bacteria was not altered by antibiotic treatment, but qualitative variations were observed. Tylosin treatment determined a significant decrease of α- and β-diversity indices and a strong depletion of the rectum population (lactobacilli and bifidobacteria) while favoring the ileum microorganisms (Gilliamella, Snodgrassella, and Frischella spp.). Major changes were also observed in honeybees treated with sulfonamides, with a decrease in Bartonella and Frischella core taxa and an increase of Bombilactobacillus spp. and Snodgrassella spp. The present study also shows an important effect of tetracycline that is focused on specific taxa with minor impact on alfa and beta diversity. Monitoring of antibiotic resistance genes confirmed that honeybees represent a great reservoir of tetracycline resistance genes. Tetracycline and sulfonamides resistance genes tended to increase in the gut microbiota population upon antibiotic administration. IMPORTANCE This study investigates the impact of the three most widely used antibiotics in the beekeeping sector (oxytetracycline, tylosin, and sulfonamides) on the honeybee gut microbiota and on the spread of antibiotic resistance genes. The research represents an advance to the present literature, considering that the tylosin and sulfonamides effects on the gut microbiota have never been studied. Another original aspect lies in the experimental approach used, as the study looks at the impact of veterinary drugs and feed supplements 24 days after the beginning of the administration, in order to explore perturbations in newly eclosed honeybees, instead of the same treated honeybee generation. Moreover, the study was not performed with cage tests but in micro-hives, thus achieving conditions closer to real hives. The study reaches the conclusion that the most common veterinary drugs determine changes in some core microbiota members and that incidence of resistance genes for tetracycline and sulfonamides increases following antibiotic treatment.

Nosema ceranae is a major pathogen in the beekeeping sector, responsible for nosemosis. This disease is hard to manage since its symptomatology is masked until a strong collapse of the colony population occurs. Conversely, no medicaments are available in the market to counteract nosemosis, and only a few feed additives, with claimed antifungal action, are available. New solutions are strongly required, especially based on natural methods alternative to veterinary drugs that might develop resistance or strongly pollute honey bees and the environment. This study aims at investigating the nosemosis antiparasitic potential of some plant extracts, microbial fermentation products, organic acids, food chain waste products, bacteriocins, and fungi. Honey bees were singularly infected with 5 × 104 freshly prepared N. ceranae spores, reared in cages and fed ad libitum with sugar syrup solution containing the active ingredient. N. ceranae in the gut of honey bees was estimated using qPCR. The results showed that some of the ingredients administered, such as acetic acid at high concentration, p-coumaric acid, and Saccharomyces sp. strain KIA1, were effective in the control of nosemosis. On the other hand, wine acetic acid strongly increased the N. ceranae amount. This study investigates the possibility of using compounds such as organic acids or biological agents including those at the base of the circular economy, i.e., wine waste production, in order to improve honeybee health.

Bees can be severely affected by various plant protection products (PPP). Among these, neonicotinoid insecticides are of concern as they have been shown to be responsible for extensive honeybee colonies death when released into the environment. Also, sublethal neonicotinoid doses contaminating single honeybees and their colonies (e.g. through contaminated pollen) are responsible for honeybees physiological alterations with probable implication also on microbiome functionality. Honeybees show symbiotic interactions with specific gut bacteria that can enhance the adult host performances. Among the known mechanisms, the modulation of the immune system, the degradation of recalcitrant secondary plant metabolites, pollen digestion, and hormonal signaling, are the most important functional benefits for the host honeybee. To date, few research efforts have aimed at revealing the impact of PPP on the gut microbial community of managed and wild honeybees. The majority of the existing literature relays on cage or semifield tests of short duration for research investigating neonicotinoids-gut microbiome interactions. This research wanted to unravel the impact of two neonicotinoids (i.e. imidacloprid and thiacloprid) in natural field conditions up to 5 weeks of exposure. A long-term impact of neonicotinoids on gut microbial community of honeybees was observed. The alterations affected several microbial genera and species such as Frischella spp., lactobacilli and bifidobacteria, whose shifting is implicated in intestinal dysbiosis. Long-term impact leading to dysbiosis was detected in case of exposure to imidacloprid, whereas thiacloprid exposure stimulated temporary dysbiosis. Moreover, the microbial diversity was significantly reduced in neonicotinoid-treated groups. Overall, the reported results support a compromised functionality of the gut microbial community, that might reflect a lower efficiency in the ecosystemic functionality of honeybees.

Honey bees are important pollinators of several crops and ecosystems, having a great ecological and economic value. In Europe, the restricted use of chemicals and therapeutic agents in the beekeeping sector has stimulated the search for natural alternatives with a special focus on gut symbionts. The modulation of the gut microbiota has been recognised as a practical and successful approach in the entomological field for the management of insect-related problems. To date, only a few studies have investigated the effect of bacterial supplementation on the health status of colonies, colony productivity and gut symbionts. To this purpose, a preparation of sugar syrup containing bifidobacteria and lactobacilli isolated from bee gut was sprayed on the frames of an apiary located in open field once a week for four weeks. Treated and control hives were monitored for two months for brood extension, honey and pollen harvest. The presence of beneficial gut microorganisms within bee gut was investigated with denaturing gradient gel electrophoresis and next generation sequencing. The administered bacteria led to a significant increase of brood population (46.2%), pollen (53.4%) and harvestable honey in honey supers (59.21%). Analysis of the gut microbiota on the new generation of bees in treated hives showed an increase in relative abundance of Acetobacteraceae and Bifidobacterium spp., which are known to be involved in bee nutrition and protection.

In order to meet the increasing demand for food quality and safety, the control of pathogenic microorganisms from “farm to fork” is a continuous challenge. This challenge has become more important due to changes in animal production, product processing and distribution, new food habits, higher numbers of consumers at risk for infection and increased awareness. This review is focused on the use of protective and probiotic cultures as “natural” intervention measures to control and prevent the transmission of pathogens along the food chain and on the most used technologies to produce these cultures at the large scale.

Environmental and hereditary factors, together with lifestyle, are important factors in colon cancer development. Considering the increasing incidence of this disease, especially in the developed western world, the last decade has seen much attention directed towards understanding possible prevention strategies. Efforts to study the intestinal microbiota and its interaction with the host have underlined that disbiosis in colonic bacterial composition is a risk factor for colon cancer. Modulation of the composition of intestinal microbiota through the use of probiotic, prebiotic and synbiotic products could therefore represent a strategy for prevention of cancer development. The mechanisms underlying the probiotic-prebiotic anticarcinogenic effect involve a combination of events: e.g. binding of mutagens, suppression of bacteria that convert pro-carcinogens into carcinogens, immune system stimulation, and a reduction in the level of certain intestinal bacterial enzymes that promote carcinogen formation.

Nosema ceranae is a widespread microsporidium of European honeybee Apis mellifera L. affecting bee health. The ban of Fumagillin-B (dicyclohexylammonium salt) in the European Union has driven the search for sustainable strategies to prevent and control the infection. The gut microbial symbionts, associated to the intestinal system of vertebrates and invertebrates and its impact on host health, are receiving increasing attention. In particular, bifidobacteria and lactobacilli, which are normal inhabitants of the digestive system of bees, are known to protect their hosts via antimicrobial metabolites, immunomodulation and competition. In this work, the dietary supplementation of gut bacteria was evaluated under laboratory conditions in bees artificially infected with the parasite and bees not artificially infected but evidencing a low natural infection. Supplemented bacteria were selected among bifidobacteria, previously isolated, and lactobacilli, isolated in this work from healthy honeybee gut. Four treatments were compared: bees fed with sugar syrup (CTR); bees fed with sugar syrup containing bifidobacteria and lactobacilli (PRO); bees infected with N. ceranae spores and fed with sugar syrup (NOS); bees infected with N. ceranae and fed with sugar syrup containing bifidobacteria and lactobacilli (NP). The sugar syrup, with or without microorganisms, was administered to bees from the first day of life for 13 days. N. ceranae infection was carried out individually on anesthetised 5-day-old bees. Eight days after infection, a significant (P

L’invenzione concerne una composizione e un metodo per promuovere la crescita di piante erbacee, in particolare piante erbacee comprese in tappeti erbosi per uso sportivo o ornamentale e in terreni agricoli, e favorire un accumulo di sostanza organica nel terreno in cui vivono e crescono le suddette piante erbacee.

Background: Fusarium head blight (FHB) is a severe disease caused by different Fusarium species, which affects a wide range of cereal crops, including wheat. It determines from 10 to 30 % of yield loss in Europe. Chemical fungicides are mainly used to reduce the incidence of FHB, but low environmental impact solutions are looked forward. Applications of soil/rhizobacteria as biocontrol agents against FHB in wheat are described in literature, whereas the potential use of lactobacilli in agriculture has scarcely been explored. Results: The aim of this work was to study the inhibitory effect of two bacterial strains, Lactobacillus plantarum SLG17 and Bacillus amyloliquefaciens FLN13, against Fusarium spp. in vitro and to assess their efficacy in field, coupled to the study of the microbial community profile of wheat seeds. Antimicrobial assays were performed on agar plates and showed that the two antagonistic strains possessed antimicrobial activity against Fusarium spp. In the field study, a mixture of the two strains was applied to durum wheat i) weekly from heading until anthesis and ii) at flowering, compared to untreated and fungicide treated plots. The FHB index, combining both disease incidence and disease severity, was used to evaluate the extent of the disease on wheat. A mixture of the two microorganisms, when applied in field from heading until anthesis, was capable of reducing the FHB index. Microbial community profile of seeds was studied via PCR-DGGE, showing the presence of L. plantarum SLG17 in wheat seeds and thus underlining an endophytic behavior of the strain. Conclusions: L. plantarum SLG17 and B. amyloliquefaciens FLN13, applied as biocontrol agents starting from the heading period until anthesis of wheat plants, are promising agents for the reduction of FHB index.

CONTENTS 16.1 Bifidobacteria and Their Use as Probiotics....................................................................................291 16.2 Dietary Flavonoid Bioconversion by Bifidobacteria.....................................................................293 16.2.1 Dietary Flavonoids...........................................................................................................293 16.2.2 Role of Bifidobacteria in Increasing the Biological Activity of Dietary Flavonoids.......295 16.3 Folate Production by Bifidobacteria..............................................................................................296 16.3.1 Structure of Folates..........................................................................................................296 16.3.2 Sources.............................................................................................................................298 16.3.3 Folate Production by Bifidobacteria.................................................................................298 16.4 Cancer Prevention Activities by Bifidobacteria............................................................................299 16.4.1 Cancer and Intestinal Microbiota.....................................................................................299 16.4.2 Bifidobacteria and Anticarcinogenic Activities...............................................................301 16.4.2.1 Binding/Absorption of Carcinogens.................................................................301 16.4.2.2 Modulation of Intestinal Bacterial Enzymes....................................................301 16.4.2.3 Effects on Oxidative Stress...............................................................................302 16.4.2.4 Production of Beneficial Compounds...............................................................302 References...............................................................................................................................................303

Characterization of strains for taxonomic purposes. The current methodologies used for the characterization of a strain for taxonomic purpose are a combination of those considered traditional and the new one that have been developed recently. In case of strains belonging to novel taxa the characterization needs to provide data for their allocation within the hierarchical framework laid down by the Bacteriological Code. The information on the strain should be as complete as possible and should include the location and the environment peculiarity from which the strain was isolated. Genetic and phenotypic methods are used for characterization. Genotypic criteria refers to sequence analysis of the 16S rRNA gene, DNA-DNA hybridization, DNA base composition, sequencing of the housekeeping genes. Phenotypic criteria typically comprises parameters such as cell shape, colony morphology, biochemical properties, pH and temperature optima etc. In addition, chemotaxonomy is in essence a part of the phenotypic characterization. Characterization of strains for probiotics applications. Functional characterization of probiotic strains have been performed in our laboratory regarding humans , animals and plants. Three examples are reported. The application to humans aimed at the characterization of Bifidobacterium strains capable of inhibiting pathogens typical of the infant gastrointestinal tract. After a preliminary screening regarding the antimicrobial properties against coliforms, Escherichia coli, Salmonella enteriditis, and Clostridium difficile strains, the most interesting Bifidobacterium strains were characterized for cytotoxic effects, adhesion to gut epithelium cell lines and for their ability to stimulate gut health by increasing the metabolic activity and the immune response of epithelial cells. The examination of all these features allowed the identification of three Bifidobacterium breve strains and a Bifidobacterium longum subsp. longum strain as potential probiotics for the treatments of enteric disorders in newborns such as infantile colics. The application to animals regarded the characterization of probiotic strains for the reduction of Campylobacter spp. levels in poultry aimed at increasing the safety of poultry meat. Within this work, a synbiotic mixture was developed, composed of a galactooligosaccharide combined with a probiotic Bifidobacterium strain (B. longum subsp. longum PCB133), characterized for its antimicrobial activity against C. jejuni. A significant reduction of C. jejuni in poultry feces was obseved in the treated chickens. The application to plants focused on the inoculation of microorganisms to improve plant growth and root development of perennial ryegrass. A microorganism-based commercial product was used to amend hydroponically grown Lolium perenne L. and results compared with the use of the same filtered product, a phytohormone solution and an untreated control. The main microbial groups present in the product were identified, evidencing bacterial and yeast species, including several Lactobacillus strains. The microbial composition of sand and root samples was investigated by PCR-DGGE, showing different profiles in the different treatments. Inoculated lactobacilli were found to be able to colonize plant roots. Plants treated with the product showed an increased resistance to tearing out with respect to other treatments and roots were longer with respect to the control.

Introduction. With the ban of dietary antimicrobial agents in animal farms, the use of probiotics, prebiotics or synbiotics to reduce pathogen load has attracted a great attention. In particular, Campylobacter jejuni has emerged as a leading bacterial cause of food-borne gastroenteritis in humans, mainly due to poultry food chain and poultry products. In a recent work a synbiotic mixture were administered for 15 days to young broilers and qPCR revealed an increase of the beneficial bacteria (i.e. bifidobacteria, lactobacilli) and a reduction of C. jejuni. In the current study the microbial gut community profiles and C. jejuni were evaluated in treated and untreated broilers by PCR-DGGE analysis to better understand the impact of the tested synbiotic product and support qPCR results. Methods. The synbiotic product was composed by the microencapsulated B. longum subsp. longum PCB133 (>109 UFC/g) and a galactooligosaccharide (3% w/w). It was administered to broilers for 15 days directly mixed to normal feed; faecal samples were collected three times along the trial (t0, t15, t20). DNA extracted from faecal samples was analyzed with DGGE technique after amplification with universal primers and with Campylobacter-specific primers (fla-DGGE). Obtained profiles analysed using the GelCompareII software package version 6.1 (Applied Maths, Kortrijk, Belgium). After normalization, a cluster analysis was performed using the Pearson similarity coefficient and unweighted pair group mathematical average (UPGMA) clustering algorithm. Results. DGGE profiles provided evidence that synbiotic consumption did not alter microbial profiles in the fecal microbiota of both untreated and treated animals; nevertheless a shift in the faecal microbiota of both groups was observed, due probably to animal growth and development. Moreover, DGGE results showed that the band corresponding to the administered probiotic strain PCB133 was detected in all treated animals and was persistent after the wash-out period. Preliminary fla-DGGE analysis underlined the effectiveness of the treatment on naturally infected animals. Discussion. The experimental study highlights the positive effect of the synbiotic approach in broiler chickens, confirming the efficacy of the probiotic PCB133 against C. jejuni without perturbation of gut microbial community.

Lactic Acid Bacteria [LAB] are worldwide known as probiotic microorganisms. One of the most important probiotic feature is their strong activity against a wide range of harmful bacteria/fungi (Gaggìa et al., 2011). Production of organic acids and production of antagonistic compounds are the main mechanisms described nowadays. This work focuses on the in vitro selection of LAB strains isolated from environmental sources against Fusarium genus. Fusarium is a well distributed genus of filamentous fungi affecting plant, animal and human health. In the agricultural field, Fusarium is one of the most important plant pathogenic genera, with a record of devastating infections in various economically important plants. In this study, in vitro antagonistic effects of 18 isolates of Lactobacillus spp. were evaluated against F. oxysporum f. sp. lactucae, agent of tracheofusariosis in Lactuca sativa, F. culmorum and F. graminearum, agents of severe diseases in cereals. Fungal inhibition was performed using spot agar test in a triplicate assay (Magnusson et al., 2003). Overnight cultures of LAB were inoculated as spot on MRS agar plates and allowed to grow at 30°C for 24 h. The plates were then overlaid with 10 ml of PDA containing 104 mould spores/ml. After 48-72 h of aerobic incubation at 25°C, the zone of inhibition was measured. All LAB strains (except one) showed antifungal activity against all Fusarium species (inhibition halo between 23.7- 65.3 mm). The highest halos were obtained against F. graminearum on which 50% of the LAB strains showed a total growth inhibition of the colony. These results are probably related to the high amount of organic acids produced by LAB strains but it can also be hypothesized the presence of antifungal compounds. In vivo application of the most successful LAB strains is foreseen towards wheat and lettuce to confirm in vitro results.

Bifidobacteria are an important commensal group of the gut microbiota of animals. They belong to key bacterial groups playing symbiotic and probiotic role promoting health activities for the host. The distribution of bifidobacterial species in non human primates is currently poorly investigated. The occurrence of bifidobacterial species in animals is characterized by a peculiar trend where there is a cluster of species typical for non primates animals and a different cluster typical of man (human primates). Recently 5 new bifidobacterial species have been isolate from non-human primates such as new world monkeys (common marmoset, Callithrix jacchus L., and red-handed tamarin, Saguinus midas L.). Due to the novelty of these species they cannot be assigned to any bifidobacterial species cluster. Studies currently under development in our lab show that bifidobacterial isolates from Orangutan and Chimpanzee belong to human primates bifidobacteria cluster: this is in agreement with the strict evolutionary link between hominoids monkey such as Orangutan and Chimpanzee and humans. The aim of the present study is to investigate the biodiversity of bifidobacterial species in primates more distant to human from an evolutionary point of view such as common marmoset, New World monkey, in order to establish where does the bifurcation between the two bifidobacterial cluster can occur. Fecal samples were collected from 5 infants of common marmoset. Colony counts and isolation of bifidobacteria were performed in mTPY (modified TPY) as describe by Rada & Petr (2000). The isolates were tested for the activity of fructose-6-phosphate phosphoketolase (F6PPK) according to Biavati and Mattarelli (2012). Rep-PCR using primer BOXAR1 (Masco et al., 2009) was conducted and the isolates were grouped using R software (Ishii et al., 2009). For the species identification, the 16S rRNA gene amplification and sequencing of one representative strain from each obtained group were performed. Bifidobacteria were isolated from all sampled animals. The faecal Bifidobacterium spp. counts ranged from 9.15 to 9.48 log10 CFU/g in infant common marmosets.. The analysis of BOX profiles reveled richness of species which, basing on the rep-PCR analysis, formed 29 groups. Sequencing analysis of 16S rRNA gene are currently on going and partial results revealed that one of the most representative strains is a novel species: it is not closely related to known Bifidobacterium species as its high sequencing similarity shared 96% to B. scardovii while values

Use of biological control agents, such as plant growth promoting rhizobacteria (PGPR), and Lactic Acid Bacteria (LAB) can be a suitable approach in control of disease in vegetable crops to reduce and limit the widespread use of chemicals. PGPR, such as Pseudomonas and Bacillus strains, are the major root colonizers that can elicit plant defense and compete with pathogens. Lactic Acid Bacteria [LAB] are worldwide known as probiotic microorganisms and one of the most important probiotic feature is their activity against a wide range of harmful bacteria/fungi. Their use as “effective microorganisms” is increasing as active components of soil amendments for sustainable agriculture. This work focuses on the in vitro screening of 32 PGPR and LAB strains isolated from environmental sources against Fusarium spp, Sclerotinia spp. and Rhizoctonia spp which are widely distributed fungi affecting plant, animal and human health. In the agricultural field, they are considered important plant pathogenic fungi, with a record of devastating infections in various economically important crop plants. Fungal inhibition was performed using spot agar test in a triplicate assay with different conditions for LAB and PGPR. All LAB strains (except one) showed an antifungal activity against all Fusarium species tested (inhibition halo between 23.7- 65.3mm); the highest halos were obtained against F. graminearum on which 50% of the strains showed a total growth inhibition of the colony. Among PGPR, preliminary results showed a strong inhibition by some B. subtilis strains against all fungi tested. These results are promising for the formulation of a commercial product that could be used for a wide spectrum of plant crops. In vivo application with the most successful strains is foreseen towards wheat and lettuce to confirm in vitro results.

Turfgrasses are not only designed for recreation activities, but they also provide beneficial environmental effects and positively influence the human wellness. Their major problems are predisposition to tearing out and microbial diseases. The aim of this study was to investigate whether the inoculation of microorganisms can be effective to improve plant growth and root development of perennial ryegrass, to evaluate new sustainable practice for green preservation. A microorganism-based commercial product was used to amend hydroponically grown Lolium perenne L. and results compared with the use of the same filtered product, a phytohormone solution and an untreated control. Plants were grown for five weeks, shoots cut and measured at one-week interval and, at the end, roots were measured for length and weight. Shoot resistance to tearing out was also tested. Moreover, the main microbial groups present in the product were characterized and the microbial profile of sand and root samples was investigated by PCR-DGGE. The plants treated with the product showed an increased resistance to tearing out with respect to other treatments and roots were longer with respect to the control. Microbial analyses of the product evidenced bacterial and yeast species with plant growth promoting activity, such as Stenothrophomonas maltophilia, Candida utilis and several Lactobacillus species. Some Lactobacillus strains were also found to be able to colonize plant roots. In conclusion, the treatment with microorganisms has a great potential for the maintenance and increased performance of turfgrass surfaces.