REFLECTIONS OF ECOSYSTEM SERVICES ON AGRICULTURAL-SOIL PHYLOGENETIC AND FUNCTIONAL DIVERSITY OF PROKARYOTES Polymerase chain reaction (PCR) based approaches in prokaryotic ecology

Il suolo è un ambiente complesso che è alla base di molteplici servizi ecosistemici per la produzione agricola. Questa complessità è riflessa nella composizione e nelle funzioni degli organismi microbici coinvolti. Lo scopo di questa tesi è stato quello di esplorare le risposte della comunità microbica all’intervento dell’uomo in suoli agricoli. Lo studio ha coinvolto suoli di comune origine, modificatisi negli ultimi 6-700 anni a causa di differente uso e gestione agronomica del suolo. Per ottenere questo risultato, il DNA del suolo è stato analizzato con tecniche di sequenziamento avanzato. I risultati indicano che i suoli disturbati sono più diversi rispetto ai suoli naturali. I fattori che influenzano la comunità microbica sono il disturbo, la disponibilità di nutrienti e la dormienza microbica. In un altro esperimento, a carico di batteri ammonio ossidanti, si sono studiati gli effetti di diversi stress come l’umidità e la concentrazione di zinco nel suolo o la presenza di pesticidi nella lettiera. In tutte queste situazioni i batteri hanno mostrato una ridondanza e una variabilità che permettono di rispondere agli stress. In conclusione i risultati di questa tesi dimostrano che l’intervento umano è responsabile nel determinare la struttura e le funzioni della comunità procariotica dei suoli.Soil is a complex environment comprising the basis for several ecosystem services, with many of them being connected to agricultural production. This complexity is reflected on the composition and functions of the hosted microbial life mainly responsible for the acquired services. Aim of the described studies was to explore microbial community responses to ecosystem services related human intervention in agricultural soils. Total prokaryotic diversity was studied in soils of common origin, which diverged in properties during the late 6-7 centuries due to different land use and management. For achieving this, related DNA markers were screened with high throughput sequencing. Cultivated environments had increased diversity compared to more natural soils. Factors potentially affecting the microbial community structure were: soil disturbance events; available nutrients; and microbial dormancy. In a second approach, ammonia oxidizing prokaryotes were used as biomarkers for studying stress effects caused by humidity and increased zinc concentrations and also the presence of organic pesticides in soil and litter respectively. In both referred cases the studied microbial guilds responded to the applied stresses showing strain or taxon level functional redundancy potentials, and tolerance variability. Overall, results show that human intervention is determining for the prokaryotic structure and functions in agricultural soils

Self-care management, knowledge, self-efficacy for medication and Quality of Life in chronic heart failure patients

Background: The management of Heart Failure (HF) is the cornerstone for the appropriate therapy and the knowledge obtained concerning the disease is known to affect self-care behaviours. Aim: The aim was to test the association between self-care management, knowledge, self-efficacy for medication and quality of life in chronic HF patients.Material and Method: A cross-sectional study was conducted and four questionnaires were used: Atlanta Heart Failure Knowledge Test (AHFK), Minnesota Living with Heart Failure (HMLHFQ), Self-Efficacy for Appropriate Medication Use Scale (SEAMS), European Heart Failure Self-care Behaviour Scale (EHFScBS-9). Multiple linear regression analysis was performed to identify independently associated variables with study subscales.Results: The sample consisted of 122 Heart Failure patients (102 men-20 women) with mean age 67.1 years (SD=12.3). Scores on Fluid and sodium management, Physical activity and recognition of deteriorating symptoms were  significantly greater  as compared with scores on Adhering to recommendations (p<0.001), indicating that the needs concerning Adhering to recommendations were greater.  Linear regression analyses showed that score on knowledge-test and New York Heart Association (NYHA) class were predictive of medication adherence factors. Better knowledge was associated with better medication adherence, while worse scores were found in subjects with NYHA class III or IV as compared to those with NYHA class I or II. Additionally, Knowledge and years of Education were associated with Self-care behaviour factors (β=0.44, SE=0.08, p<0.001). Conclusions: Patients at a higher level of HF knowledge show better adherence to the Medication. Self-care behavior and life quality are improved

Using biochar for environmental recovery and boosting the yield of valuable non-food crops: the case of hemp in a soil contaminated by potentially toxic elements (PTEs)

Hemp (Cannabis sativa L.) is known to tolerate high concentrations of soil contaminants which however can limit its biomass yield. On the other hand, organic-based amendments such as biochar can immobilize soil contaminants and assist hemp growth in soils contaminated by potentially toxic elements (PTEs), allowing for environmental recovery and income generation, e.g. due to green energy production from plant biomass. The aim of this study was therefore to evaluate the suitability of a softwood-derived biochar to enhance hemp growth and promote the assisted phytoremediation of a PTE-contaminated soil (i.e., Sb 2175 mg kg-1; Zn 3149 mg kg-1; Pb 403 mg kg-1; and Cd 12 mg kg-1). Adding 3% (w/w) biochar to soil favoured the reduction of soluble and exchangeable PTEs, decreased soil dehydrogenase activity (by ~2.08-fold), and increased alkaline phosphomonoesterase and urease activities, basal respiration and soil microbial carbon (by ~1.18-, 1.22-, 1.22-, and 1.66-fold, respectively). Biochar increased the abundance of selected soil culturable microorganisms, while amplicon sequencing analysis showed a positive biochar impact on α-diversity and the induction of structural changes on soil bacterial community structure. Biochar did not affect root growth of hemp but significantly increased its aboveground biomass by ~1.67-fold for shoots, and by ~2-fold for both seed number and weight. Biochar increased the PTEs phytostabilisation potential of hemp with respect to Cd, Pb and Zn, and also stimulated hemp phytoextracting capacity with respect to Sb. Overall, the results showed that biochar can boost hemp yield and its phytoremediation effectiveness in soils contaminated by PTEs providing valuable biomass that can generate profit in economic, environmental and sustainability terms

Διοξίνες και φουράνια: Επιπτώσεις στην ανθρώπινη υγεία

Σκοπός της παρούσας Μεταπτυχιακής Διπλωματικής Εργασίας είναι η καταγραφή και η ανάλυση των σημαντικότερων χαρακτηριστικών των διοξινών και των φουρανίων, καθώς και των επιπτώσεών τους στην ανθρώπινη υγεία, ούτως ώστε να αναδειχθούν ορισμένες δυσμενείς συνέπειες των ανθρωπογενών δραστηριοτήτων. Οι πολυχλωριωμένες διβενζο-παρα-διοξίνες (PCDDs) και τα πολυχλωριωμένα διβενζοφουράνια (PCDFs) απαρτίζουν μία ομάδα τοξικών χημικών ρυπαντών, γνωστών ως ανθεκτικών οργανικών ρύπων (POPs). Τα οργανικά αυτά μόρια, λοιπόν, σχηματίζονται κυρίως ως παραπροϊόντα βιομηχανικής διεργασίας, χημικής καύσης, καύσης ξύλου και αποτέφρωσης απορριμμάτων, τα οποία δε, είναι άνευ χρησιμότητας για τον άνθρωπο. Οι εν λόγω περιβαλλοντικοί ρυπαντές είναι πανταχού παρόντες, καθώς ανιχνεύονται, έστω και σε ιχνοποσότητες, στον αέρα, στα ύδατα, στο έδαφος, στα ιζήματα, στην πανίδα, στη χλωρίδα, στα στερεά απόβλητα και στα λύματα. Επιπλέον, ως λιπόφιλες ουσίες έχουν την ικανότητα να απορροφώνται από τον λιπώδη ιστό με αποτέλεσμα να βιοσυσσωρεύονται στα ζώα και ως εκ τούτου, μέσω της τροφικής αλυσίδας που αποτελεί την κυριότερη πηγή έκθεσης, μπορούν να μεταφέρονται στον άνθρωπο. Εν ολίγοις, η άμεση και η έμμεση έκθεση του ανθρώπου σε αυτούς τους ύπουλους ρυπαντές είναι συνεχής, αναπόφευκτη και ξεκινάει από την in utero ζωή. Ωστόσο, η έκθεση, εκτός από περιβαλλοντική, μπορεί να είναι επαγγελματική και ατυχηματική, ενώ ακόμη υπάρχει η δυνατότητα χρήσης αυτών των χημικών παραγόντων ως όπλων. Επιπροσθέτως, έχει δειχθεί ότι ορισμένα από τα ομοειδή των PCDDs και των PCDFs, των οποίων ο μηχανισμός δράσης σχετίζεται με τον υποδοχέα Ah, παρουσιάζουν υψηλή τοξικότητα με αποτέλεσμα να καταγράφονται αρνητικές επιπτώσεις στην υγεία των ανθρώπων αλλά και των ζώων. Ανάμεσα στις αρνητικές επιδράσεις αυτών των τοξικών ουσιών συγκαταλέγονται η χλωρακμή, η καρκινογένεση, οι νευροαναπτυξιακές διαταραχές, οι διαταραχές στην ανοσολογική απόκριση, οι ενδοκρινικές διαταραχές και η αναπαραγωγική δυσλειτουργία με χαρακτηριστικό παράδειγμα τον χαμηλό αριθμό σπερματοζωαρίων. Ακόμη, υφίστανται γονιδιοτοξικές επιπτώσεις αλλά και δυσμενείς επιδράσεις σε άλλα βιολογικά συστήματα, όπως είναι το καρδιαγγειακό σύστημα, το αναπνευστικό σύστημα και το γαστρεντερικό σύστημα. Τέλος, ας σημειωθεί ότι το τοξικότερο ομοειδές αυτών των οργανικών μορίων είναι η 2,3,7,8-τετραχλωρο-p-διοξίνη (TCDD), η οποία έχει χαρακτηριστεί ως καρκινογόνος ουσία για τον άνθρωπο από τον Παγκόσμιο Οργανισμό Υγείας και αποτελεί ουσία αναφοράς για τον προσδιορισμό ορισμένων τοξικολογικών δεικτών, όπως είναι το τοξικό ισοδύναμο (TEQ).The aim of the present Postgraduate Diploma Thesis is to record and analyze the most important characteristics of dioxins and furans as well as their effects on human health, in order to highlight certain adverse consequences of anthropogenic activities. Polychlorinated dibenzo-para-dioxins (PCDDs) and polychlorinated dibenzofurans (PCDFs) constitute a group of chemical pollutants, known as persistent organic pollutants (POPs). So, these organic molecules are mainly formed as by-products of industrial processes, chemical combustion, wood burning and waste incineration, which are of no use to human. These environmental pollutants are ubiquitous as they are detected, even in trace amounts, in air, water, soil, sediments, fauna, flora, solid waste and sewage. In addition, as lipophilic substances they have the ability to be absorbed by adipose tissue resulting in bioaccumulation in animals and hence, through the food chain which is the main source of exposure, they can be transferred to human. In sum, direct and indirect human exposure to these insidious pollutants is continuous, unavoidable and it begins in utero life. However, exposure can be occupational and accidental, beyond environmental, and there is the possibility of using those chemical agents as weapons. Additionally, it has been shown that some congeners of PCDDs and PCDFs, which mechanism of action is related to Ah receptor, have high toxicity resulting negative effects on human and animal health. Among the negative effects of these toxic substances are chloracne, carcinogenesis, neurodevelopmental disorders, immune system disorders, endocrine disorders and reproductive dysfunction such as in the characteristic case of low sperm count. Furthermore, there are genotoxic effects as well as adverse effects on other biological systems, such as the cardiovascular system, the respiratory system and the gastrointestinal system. Finally, it should be noted that the most toxic congener of these organic molecules is 2,3,7,8-tetrachloro-p-dioxin (TCDD), which has been classified as a human carcinogen by World Health Organization and it is a reference substance for the determination of certain toxicological indices, such as the toxic equivalent (TEQ)

Pesticide researchers face formidable challenges: Annual Meeting Report of the Mediterranean Group of Pesticide Research, Piacenza, Italy, 13 and 14 November 2008

The Mediterranean Group of Pesticide Research (MGPR) is a scientific society of people committed to research concerning effects of pesticides on the environment and human health. A main task of the MGPR is to contribute to evaluation and dissemination of relevant scientific information, which includes novel analytical methods for pesticide-residue analysis, updates about new pesticides or new application methods, evaluation of pesti- cides and pesticide-application methods in the context of sustainable agriculture and the relevant legislation. Another strong point of MGPR is encouragement of a collaborative spirit among its members through the formation of working groups in order to accomplish its mission, so annual meetings serve as reference points for not only information but also collaboration among scientists in the field of pesticides research. Update on the work carried out by the members of the MGPR and other researchers of the field was given during the annual meeting that took place in Piacenza, Italy, 13 and 14 November 2008. Activities included presentations, a poster session, a technical update on technical advances at the host institute (Universita Cattolica, Piacenza, Italy) and social events

Different factors are operative in shaping the epiphytic grapevine microbiome across different geographical scales: Biogeography, cultivar or vintage?

Abstract Introduction The composition of the grapevine microbiome is controlled by a range of factors all contributing to the establishment of microbial terroir. Most studies have focused on the grape must microbiome, while less is known about the assemblage mechanisms of the epiphytic grapevine microbiome. We aimed to disentangle the role of geographic location, cultivar and vintage on the composition of the epiphytic fungal and bacterial communities of grapevine: within (regional scale) and across (national scale) viticultural zones in Greece. We hypothesised that: (i) the influence of all three factors varies at different geographical scales; (ii) fungi and bacteria respond differently to the factors shaping the grapevine microbiome. Materials and Methods We analysed leaves and berries collected from Greek cultivars (i) Vidiano and Agiorgitiko located in three geographically distant regions (from 100 to >600 km), (ii) Roditis and Sideritis from different terroir units of the viticultural zone of Aigialeia. Results Amplicon sequencing analysis identified strong regional signatures on the epiphytic microbiome across viticultural zones, whereas cultivar becomes a strong determinant at the viticultural zone scale. Fungal communities were more responsive to all studied factors compared to bacterial communities. Differential abundance (DA) analysis identified dominant fungal (Alternaria, Cladosporium, Nigrospora, Aureobasidium, Vishniacozyma) and bacterial (Sphingomonas, Masillia, Streptococcus, Staphylococcus, Enterobacterales) amplicon sequence variants (ASVs) exhibiting geographical and cultivar‐specific patterns. Conclusion Our data suggest that different factors are operative in shaping the epiphytic grapevine microbiome at different geographical scales with bacterial and fungal communities showing different responses to the tested structural factors

How do metal pollutant concentration and speciation affect wastewater microbial diversity and antibiotic resistance?

In this study we compared metal-induced cross-resistance to antibiotics in urban wastewater, whilst paying particular attention to issues of metal speciation. Municipal wastewater influent was used to establish microcosms spiked with silver (Ag), copper (Cu), or zinc (Zn) at concentrations from 1-10000 μM. All treatments were sampled for analysis after 1 and 7 day's incubation. X-ray absorption spectroscopy (a powerful, synchrotron-based structural technique) was used to directly investigate Ag, Cu, and Zn speciation. Diffusive Gradients in Thin Film (DGT), filtration, and acid digestion based methods were used to further interrogate metal chemistry and solubility. Partial 16S rRNA amplicon sequencing was used to assess bacterial diversity, and real-time PCR to measure relative abundance of marker genes for metal (Ag, Cu, Zn) resistance, antibiotic-resistance, and horizontal-gene-transfer (HGT), and to confirm interesting trends indicated by the sequencing data. Multivariate statistical analysis of bacterial community responses to Ag, Cu and Zn showed that exposure time, metal concentration gradient, and metal speciation were all significant factors associated with measured changes. Ag/Cu/Zn sulfides comprised a significant proportion of the metal speciation profiles at low-medium range concentrations, whereas Ag-chloride, metal-phosphate and organic bound species were more important at higher concentrations. Maximum likelihood phylogenies of dominant OTUs indicated strong metal induced selection of potential human pathogens (e.g. Acinetobacter spp., Arcobacter spp.); this was confirmed by targeted qPCR (e.g. for A. baumanii, A. butzlerii). These results demonstrate that wastewater metal pollutants induce significant increases in microbial resistance and HGT potential. Importantly, metal speciation is frequently overlooked in studies investigating metal-induced microbial effects but can significantly affect microbial community selection and function

Silver Toxicity Thresholds for Multiple Soil Microbial Biomarkers

Material flow analysis shows that soil is a key repository for silver (Ag) from (nano)silver-functionalized consumer products, but the potential effects of Ag toxicity, via Ag+ release, on soil microbial communities and their ecosystem services remains largely unknown. We examined the responses of multiple microbial biomarkers to increasing Ag+ doses (nine concentrations, 0-2000 mg kg-1) in nine different soils representing a wide range of soil properties. Analyses included substrate-induced microbial respiration, nine different soil enzyme activities, and quantification of bacterial 16S-rRNA (SSU) and fungal intergenic spacer (ITS) copies. The resulting half-maximal effective concentrations (EC50) for Ag ranged from ∼1 to >500 mg kg -1 and showed soil-specific responses, including some hormesis-type responses. Carbon cycle-associated enzyme activities (e.g., cellobiohydrolase, xylosidase, and α/β-glucosidase) responded similarly to Ag. Sulfatase and leucine-aminopeptidase activities (linked to the sulfur and nitrogen cycles) were the most sensitive to Ag. Total organic carbon, and to a lesser extent pH, were identified as potentially useful response predictors, but only for some biomarkers; this reflects the complexity of soil Ag chemistry. Our results show Ag toxicity is highly dependent on soil characteristics and the specific microbial parameter under investigation, but end point redundancies also indicated that representative parameters for key microbial functions can be identified for risk assessment purposes. Sulfatase activity may be an important Ag toxicity biomarker; its response was highly sensitive and not correlated with that of other biomarkers

Host Species Determines the Composition of the Prokaryotic Microbiota in Phlebotomus Sandflies

Phlebotomine sandflies are vectors of the humans&rsquo; and mammals&rsquo; parasite Leishmania spp. Although the role of gut microbiome in the biological cycle of insects is acknowledged, we still know little about the factors modulating the composition of the gut microbiota of sandflies. We tested whether host species impose a strong structural effect on the gut microbiota of Phlebotomus spp. Sandflies were collected from the island of Leros, Greece, and classified to P. papatasi, P. neglectus, P. tobbi, and P. similis, all being negative to Leishmania spp. The prokaryotic gut microbiota was determined via 16S rRNA gene amplicon sequencing. Phlebotomus species supported distinct microbial communities (p &lt; 0.001). P. papatasi microbiota was the most distinct over-dominated by three Spiroplasma, Wolbachia and Paenibacillus operational taxonomic units (OTUs), while another Wolbachia OTU prevailed in P. neglectus. Conversely, the microbiota of P. tobbi and P. similis was composed of several less dominant OTUs. Archaea showed low presence with the dominant OTUs belonging to methanogenic Euryarcheota, ammonia-oxidizing Thaumarcheota, and Nanoarchaeota. We provide first insights into the composition of the bacterial and archaeal community of Phlebotomus sandflies and showed that, in the absence of Leishmania, host genotype is the major modulator of Phlebotomus sandfly gut microbiota