Space-temporal dynamics of limnological variables in a floodplain wetland located at Ribeira do Iguape Valley (São Paulo, Brazil) at different hydrological periods

AIM: The main goal of this research was to investigate the influence of the hydrological pulses on the space-temporal dynamics of physical and chemical variables in a wetland adjacent to Jacupiranguinha River (São Paulo, Brazil); METHODS: Eleven sampling points were distributed among the wetland, a tributary by its left side and the adjacent river. Four samplings were carried out, covering the rainy and the dry periods. Measures of pH, dissolved oxygen, electrical conductivity and redox potential were taken in regular intervals of the water column using a multiparametric probe. Water samples were collected for the nitrogen and total phosphorus analysis, as well as their dissolved fractions (dissolved inorganic phosphorus, total dissolved phosphorus, ammoniacal nitrogen and nitrate). Total alkalinity and suspended solids were also quantified; RESULTS: The Multivariate Analysis of Variance showed the influence of the seasonality on the variability of the investigated variables, while the Principal Component Analysis gave rise in two statistical significant axes, which delimited two groups representative of the rainy and dry periods. Hydrological pulses from Jacupiranguinha River, besides contributing to the inputs of nutrients and sediments during the period of connectivity, accounted for the decrease in spatial gradients in the wetland. This "homogenization effect" was evidenced by the Cluster Analysis. The research also showed an industrial raw effluent as the main point source of phosphorus to the Jacupiranguinha River and, indirectly, to the wetland; CONCLUSIONS: Therefore, considering the scarcity of information about the wetlands in the study area, this research, besides contributing to the understanding of the influence of hydrological pulses on the investigated environmental variables, showed the need for adoption of conservation policies of these ecosystems face the increase anthropic pressures that they have been submitted, which may result in lack of their ecological, social and economic functions.

Aumento de fijación de nitrógeno en diazótrofos unicelulares versus Trichodesmium tras un evento de deposición de polvo atmosférico en las Islas Canarias

[ES]En este trabajo hemos estudiado la relación entre la deposición de polvo atmosférico, y la abundancia y tasas de fijación de nitrógeno asociadas a Trichodesmium y fijadores unicelulares en las Islas Canarias. La fijación de nitrógeno asociada a los fijadores unicelulares aumentó entre el 86 y el 92% tras un evento de deposición de polvo atmosférico, mientras que la asociada a Trichodesmium disminuyó entre el 34 y el 92%. Tras el evento de deposición de polvo, aumentó la abundancia de fijadores unicelulares y la mayoría de éstos aparecieron asociados a partículas de materia orgánica. Creemos que esta estrategia les permite ligar el hierro que contiene el polvo y así poder usar este nutriente limitante.

Organismi acquatici e ambiente: meccanismi biochimici di interazione, risposta e adattamento

The research is focused on the relationship between some Mg2+-dependent ATPase activities of plasma- and mitochondrial membranes from tissues of cultured marine bivalve molluscs and potentially stressful environmental conditions, such as the exposure to contaminants both of natural origin (ammonia nitrogen, the main contaminant of aquaculture plants) and of anthropic source (alkyltins). The two filter-feeding bivalve species selected colonize different habitats: the common mussel Mytilus galloprovincialis binds to hard substrates and the Philippine clam Tapes philippinarum burrows into sea bottom sandy beds. The choice of typical species of coastal waters, extremely suitable for environmental studies due to their features of poor motility, resistance to transport and great filtering efficiency, may constitute a model to evaluate responses to contaminants of membrane-bound enzyme activities involved in key biochemical mechanisms, namely cell ionic regulation and mitochondrial energy production. In vitro and in vitro approaches have been pursued. In vitro assays were carried out by adding the contaminants (NH4Cl and alkyltins) directly to the ATPase reaction media. In vivo experiments were carried out by exposing mussels to various tributyl tin (TBT) concentrations under controlled conditions in aquaria. ATPase activities were determined spectrophotometrically according to the principles of the method of Fiske and Subbarow (1925). The main results obtained are detailed below. In Tapes philippinarum the interaction of NH4 +, the main form of ammonia nitrogen at physiological and seawater pHs, with the Na,K-ATPase and the ouabaininsensitive Na-ATPase was investigated in vitro on gill and mantle microsomal membranes. The proven replacement by NH4 +of K+ in the activation of the Na,KATPase and of Na+ in the activation of the ouabain-insensitive ATPase displayed similar enzyme affinity for the substituted cation. on the one hand this finding may represent one of the possible mechanisms of ammonia toxicity and, on the other, it supports the hypothesis that NH4 + can be transported across the plasma membrane through the two ATPases. In this case both microsomal ATPases may be involved and co-operate, at least under peculiar circumstances, to nitrogen excretion and ammonia detoxification mechanisms in bivalve molluscs. The two ATPase activities stimulated by NH4 + maintained their typical response to the glycoside ouabain, specific inhibitor of the Na,K-ATPase, being the Na++ NH4 +-activated ATPase even more susceptive to the inhibitor and the ouabain-insensitive ATPase activity activated indifferently by Na+ or NH4 + unaffected by up to 10-2 M ouabain. In vitro assays were carried out to evaluate the response of the two Na-dependent ATPases to organotins in clams and mussels and to investigate the interaction of TBT with mussel mitochondrial oligomycin-sensitive Mg-ATPase. Since no literature data were available, the optimal assay conditions and oligomycin sensitivity of mussel mitochondrial MgATPase were determined. In T. philippinarum the ouabain-insensitive Na-ATPase was found to be refractory to TBT both in the gills and in the mantle, whereas the Na,K-ATPase was progressively inhibited by increasing TBT doses; the enzyme inhibition was more pronounced in the gills than in the mantle. In both tissues of M. galloprovincialis the Na,K-ATPase inhibition by alkyltins decreased in the order TBT>DBT(dibutyltin)>>MBT(monobutyltin)=TeET(tetraethyltin) (no effect). Mussel Na-ATPase confirmed its refractorimess to TBT and derivatives both in the gills and in the mantle. These results indicate that the Na,K-ATPase inhibition decreases as the number of alkyl chains bound to tin decreases; however a certain polarity of the organotin molecule is required to yield Na,K-ATPase inhibition, since no enzyme inhibition occurred in the presence of tetraalkyl-substituted derivatives such as TeET . Assays carried out in the presence of the dithioerythritol (DTE) pointed out that the sulphhydrylic agent is capable to prevent the Na,K-ATPase inhibition by TBT, thus suggesting that the inhibitor may link to -SH groups of the enzyme complex.. Finally, the different effect of alkyltins on the two Na-dependent ATPases may constitute a further tool to differentiate between the two enzyme activities. These results add to the wealth of literature data describing different responses of the two enzyme activities to endogenous and exogenous modulators . Mussel mitochondrial Mg-ATPase was also found to be in vitro inhibited by TBT both in the gills and in the mantle: the enzyme inhibition followed non competitive kinetics. The failed effect of DTE pointed out that in this case the interaction of TBT with the enzyme complex is probably different from that with the Na,K-ATPase. The results are consistent with literature data showing that alkyltin may interact with enzyme structures with different mechanisms. Mussel exposure to different TBT sublethal doses in aquaria was carried out for 120 hours. Two samplings (after 24 and 120 hrs) were performed in order to evaluate a short-term response of gill and mantle Na,K-ATPase, ouabain-insensitive Na-ATPase and Mg-ATPase activities. The in vivo response to the contaminants of the enzyme activities under study was shown to be partially different from that pointed out in the in vitro assays. Mitochondrial Mg-ATPase activity appeared to be activated in TBTexposed mussels with respect to control ones, thus confirming the complexity of evaluating in vivo responses of the enzyme activities to contaminants, due to possible interactions of toxicants with molluscan metabolism. Concluding, the whole of data point out that microsomal and mitochondrial ATPase activities of bivalve molluscs are generally responsive to environmental contaminants and suggest that in some cases membrane-bound enzyme activities may represent the molecular target of their toxicity. Since the Na,K-ATPase, the Na-ATPase and the Mg-ATPase activities are poorly studied in marine bivalves, this research may contribute to enlarge knowledge in this quite unexplored field.

Short-term variability of picoplankton across the wind-shear region south of Gran Canaria

[EN]We have studied the short-term variability -at temporal scale of days and spatial scale of 5 km- of the hydrographic field, organic and inorganic nutrients, chlorophyll and picoplanktonic abundances, across a 40 Km section crossing a frontal system south of Gran Canaria, where anticyclonic eddies in early-stages of formation and convergent fronts have been widely reported in the past. Each cruise consisted in a 3-4 daily-repeated section, and was carried out at the same period of the year (May) during two consecutive years (2011 and 2012). The main goal of our study was to analyze the picoplankton response to short-term variability at scales not considered in regular oceanographic samplings, even in regions with complex hydrographic fields.

New approaches to open problems in gene expression microarray data

In the past decade, the advent of efficient genome sequencing tools and high-throughput experimental biotechnology has lead to enormous progress in the life science. Among the most important innovations is the microarray tecnology. It allows to quantify the expression for thousands of genes simultaneously by measurin the hybridization from a tissue of interest to probes on a small glass or plastic slide. The characteristics of these data include a fair amount of random noise, a predictor dimension in the thousand, and a sample noise in the dozens. One of the most exciting areas to which microarray technology has been applied is the challenge of deciphering complex disease such as cancer. In these studies, samples are taken from two or more groups of individuals with heterogeneous phenotypes, pathologies, or clinical outcomes. these samples are hybridized to microarrays in an effort to find a small number of genes which are strongly correlated with the group of individuals. Eventhough today methods to analyse the data are welle developed and close to reach a standard organization (through the effort of preposed International project like Microarray Gene Expression Data -MGED- Society [1]) it is not unfrequant to stumble in a clinician's question that do not have a compelling statistical method that could permit to answer it.The contribution of this dissertation in deciphering disease regards the development of new approaches aiming at handle open problems posed by clinicians in handle specific experimental designs. In Chapter 1 starting from a biological necessary introduction, we revise the microarray tecnologies and all the important steps that involve an experiment from the production of the array, to the quality controls ending with preprocessing steps that will be used into the data analysis in the rest of the dissertation. While in Chapter 2 a critical review of standard analysis methods are provided stressing most of problems that In Chapter 3 is introduced a method to adress the issue of unbalanced design of miacroarray experiments. In microarray experiments, experimental design is a crucial starting-point for obtaining reasonable results. In a two-class problem, an equal or similar number of samples it should be collected between the two classes. However in some cases, e.g. rare pathologies, the approach to be taken is less evident. We propose to address this issue by applying a modified version of SAM [2]. MultiSAM consists in a reiterated application of a SAM analysis, comparing the less populated class (LPC) with 1,000 random samplings of the same size from the more populated class (MPC) A list of the differentially expressed genes is generated for each SAM application. After 1,000 reiterations, each single probe given a "score" ranging from 0 to 1,000 based on its recurrence in the 1,000 lists as differentially expressed. The performance of MultiSAM was compared to the performance of SAM and LIMMA [3] over two simulated data sets via beta and exponential distribution. The results of all three algorithms over low- noise data sets seems acceptable However, on a real unbalanced two-channel data set reagardin Chronic Lymphocitic Leukemia, LIMMA finds no significant probe, SAM finds 23 significantly changed probes but cannot separate the two classes, while MultiSAM finds 122 probes with score >300 and separates the data into two clusters by hierarchical clustering. We also report extra-assay validation in terms of differentially expressed genes Although standard algorithms perform well over low-noise simulated data sets, multi-SAM seems to be the only one able to reveal subtle differences in gene expression profiles on real unbalanced data. In Chapter 4 a method to adress similarities evaluation in a three-class prblem by means of Relevance Vector Machine [4] is described. In fact, looking at microarray data in a prognostic and diagnostic clinical framework, not only differences could have a crucial role. In some cases similarities can give useful and, sometimes even more, important information. The goal, given three classes, could be to establish, with a certain level of confidence, if the third one is similar to the first or the second one. In this work we show that Relevance Vector Machine (RVM) [2] could be a possible solutions to the limitation of standard supervised classification. In fact, RVM offers many advantages compared, for example, with his well-known precursor (Support Vector Machine - SVM [3]). Among these advantages, the estimate of posterior probability of class membership represents a key feature to address the similarity issue. This is a highly important, but often overlooked, option of any practical pattern recognition system. We focused on Tumor-Grade-three-class problem, so we have 67 samples of grade I (G1), 54 samples of grade 3 (G3) and 100 samples of grade 2 (G2). The goal is to find a model able to separate G1 from G3, then evaluate the third class G2 as test-set to obtain the probability for samples of G2 to be member of class G1 or class G3. The analysis showed that breast cancer samples of grade II have a molecular profile more similar to breast cancer samples of grade I. Looking at the literature this result have been guessed, but no measure of significance was gived before.

Effect of multiple sources of disturbance on rocky benthic assemblages in some localities of Salento, Apulia

The first part of my work consisted in samplings conduced in nine different localities of the salento peninsula and Apulia (Italy): Costa Merlata (BR), Punta Penne (BR), Santa Cesarea terme (LE), Santa Caterina (LE), Torre Inserraglio (LE), Torre Guaceto (BR), Porto Cesareo (LE), Otranto (LE), Isole Tremiti (FG). I collected data of species percentage covering from the infralittoral rocky zone, using squares of 50x50 cm. We considered 3 sites for location and 10 replicates for each site, which has been taken randomly. Then I took other data about the same places, collected in some years, and I combined them together, to do a spatial analysis. So I started from a data set of 1896 samples but I decided not to consider time as a factor because I have reason to think that in this period of time anthropogenic stressors and their effects (if present), didn’t change considerably. The response variable I’ve analysed is the covering percentage of an amount of 243 species (subsequently merged into 32 functional groups), including seaweeds, invertebrates, sediment and rock. 2 After the sampling, I have been spent a period of two months at the Hopkins Marine Station of Stanford University, in Monterey (California,USA), at Fiorenza Micheli's laboratory. I've been carried out statistical analysis on my data set, using the software PRIMER 6. My explorative analysis starts with a nMDS in PRIMER 6, considering the original data matrix without, for the moment, the effect of stressors. What comes out is a good separation between localities and it confirms the result of ANOSIM analysis conduced on the original data matrix. What is possible to ensure is that there is not a separation led by a geographic pattern, but there should be something else that leads the differences. Is clear the presence of at least three groups: one composed by Porto cesareo, Torre Guaceto and Isole tremiti (the only marine protected areas considered in this work); another one by Otranto, and the last one by the rest of little, impacted localities. Inside the localities that include MPA(Marine Protected Areas), is also possible to observe a sort of grouping between protected and controlled areas. What comes out from SIMPER analysis is that the most of the species involved in leading differences between populations are not rare species, like: Cystoseira spp., Mytilus sp. and ECR. Moreover I assigned discrete values (0,1,2) of each stressor to all the sites I considered, in relation to the intensity with which the anthropogenic factor affect the localities. 3 Then I tried to estabilish if there were some significant interactions between stressors: by using Spearman rank correlation and Spearman tables of significance, and taking into account 17 grades of freedom, the outcome shows some significant stressors interactions. Then I built a nMDS considering the stressors as response variable. The result was positive: localities are well separeted by stressors. Consequently I related the matrix with 'localities and species' with the 'localities and stressors' one. Stressors combination explains with a good significance level the variability inside my populations. I tried with all the possible data transformations (none, square root, fourth root, log (X+1), P/A), but the fourth root seemed to be the best one, with the highest level of significativity, meaning that also rare species can influence the result. The challenge will be to characterize better which kind of stressors (including also natural ones), act on the ecosystem; and give them a quantitative and more accurate values, trying to understand how they interact (in an additive or non-additive way).

Habitatwahl und Artenzusammensetzung von Arthropodenpopulationen im urbanen Bereich am Beispiel des Rhein-Main-Ballungsraumes unter besonderer Berücksichtigung der Saltatoria

SummaryThe present thesis deals with the choosing of habitats and species structures of invertebrate populations in urban areas. The main emphasis is focused on the group of saltatoria. Samplings by stationary traps and also by net-catching were carried out from 1993 to 1997. The survey extended over the whole city area of Mainz, Germany, as well as several spots in the area of Frankfurt upon Main. Shifting calcerous sands received special consideration. For a better distinction of the surveying area, three different zones were established, 'Relicts of natural landscape', 'Traditional worked rural landscape' and 'Urban environment'. Partly significant differences in species and individual abundance between the zones showed the influence of the different cultivation on the fauna. To investigate population size and -structure as well as migration and habitat preference - especially in urban areas - capture-recapture experiments were performed on the two species Oedipoda caerulescens L. and Sphingonotus caerulans L. in 1998. Large secondary habitats, such as train stations, are getting more and more important for the survival of the highly endangered S. caerulans. Another centre of attention was the structure of invertebrate populations on class level. The results of the investigation show, that the importance of urban environment for the conservation of species and also nature has been highly underestimated so far.ZusammenfassungDie vorliegende Arbeit beschäftigt sich mit der Habitatwahl und Artenzusammensetzung von Arthropodenpopulationen im besiedelten Bereich. Der Schwerpunkt lag auf der Gruppe der Saltatoria. Über mehrere Jahre (1993-1996) wurden Fänge mit stationären Fallen sowie Kescherfänge durchgeführt. Das Untersuchungsgebiet umfaßte in erster Linie das Stadtgebiet von Mainz, darüber hinaus wurden einzelne Standorte in Frankfurt/M. und Umgebung untersucht. Ein Schwerpunkt der Untersuchungen lag auf Kalkflugsanddünen, besonders Restflächen im besiedelten Bereich. Sekundäre Sandstandorte oder vergleichbare anthropogen entstandene Böden wurden ebenfalls beprobt Das Stadtgebiet wurde aufgrund besserer Unterscheidung in drei Zonen aufgeteilt, die 'Naturlandschaftsrelikte', die 'Zone der Alten Kulturlandschaft' und die 'Urbane Landschaft'. Größtenteils signifikante Unterschiede in der Arten- und Individuenanzahl zwischen den Zonen zeigen den bedeutenden Einfluß der unterschiedlichen Bearbeitung und Kultivierung auf die Fauna. An den beiden Spezies Oedipoda caerulescens L. und Sphingonotus caerulans L. wurden Fang-Wiederfangexperimente zur Abschätzung der Populationsgröße, -struktur, Habitatwahl sowie Migration durchgeführt. Besonders für die stark gefährdete S. caerulans stellte sich die enorme Bedeutung von großflächigen Sekundärhabitaten im besiedelten Bereich, wie sie Bahnhöfe darstellen, heraus. Der zweite Schwerpunkt lag auf der Struktur von Invertebratenpopulationen im besiedelten Bereich. Sie wurden auf Ordnungsniveau erfaßt. Die Ergebnisse der Untersuchungen zeigen, daß die Bedeutung des besiedelten Bereiches für den Arten- und Biotopschutz bisher weit unterschätzt wurde.

Effetti della Salinizzazione sugli Invertebrati del suolo

Lo scopo di questa tesi è quello di valutare l'effetto della salinizzazione dei suoli sugli invertebrati edafici. Nell'ambito di questo obiettivo generale sono state effettuate due distinte attività di ricerca: una indagine sul campo e una serie di esperimenti di laboratorio. Lo studio sul campo è stato condotto nella Pineta di San Vitale (Ravenna, Italia). L'obiettivo specifico è stato quello di valutare la qualità biologica dei suoli attraverso l'analisi del popolamento dei microartropodi edafici, in relazione alla diversità del suolo e in particolare alla salinizzazione. La qualità biologica dei suoli è stata valutata mediante l'indice QBS-ar. La Pineta è stata campionata nella zona Est, più colpita da intrusione salina e nella zona Ovest dove questo fenomeno è meno evidente. I campionamenti sono stati effettuati in primavera ed estate. I risultati confermano che le caratteristiche chimico-fisiche si modificano in base al gradiente sommità dunali-depressioni interdunali. Per quanto riguarda il popolamento dei microartropodi alcune caratteristiche sono comuni alla maggior parte delle stazioni con lo stesso pedotipo. Non è stato evidenziato alcuno stress sui popolamenti attribuibule alla salinizzazione. Nel complesso, i valori di QBS-ar sono piuttosto elevati. Gli esperimenti di laboratorio sono stati finalizzata alla valutazione degli effetti combinati della salinità del suolo e della contaminazione da pesticidi (chlorpyrifos) sul lombrico Eisenia andrei. Nel complesso, i risultati indicano che effetti avversi sui lombrichi sono possibili a livelli di salinizzazione dei suoli ancora compatibili a concentrazioni di chlorpyrifos che sono piuttosto alte in confronto con i tipici risultati di campo, ma ancora compatibili con l'uso consigliato.

Patogeni e zecche vettrici: studio epidemiologico in parchi pubblici dell'Emilia Romagna

Le malattie trasmesse da zecche sono un importante problema sia per la salute animale che per quella umana e negli ultimi decenni hanno aumentato notevolmente la loro diffusione, in seguito ai cambiamenti climatici, che hanno permesso la distribuzione delle zecche in aree prima non interessate. Per tale motivo si è deciso di effettuare un’indagine sulla diffusione delle zecche e sui patogeni da loro trasmessi, mediante campionamenti sia a livello ambientale, sia su animali e umani infestati in quattro siti di tre parchi dell’Emilia Romagna, dove non risultavano precedenti segnalazioni, nelle province di Bologna e Ravenna, da Aprile a Ottobre 2010. In totale sono state raccolte 8212 zecche. Dall’ambiente sono state campionate 6734 larve, 1344 ninfe, 61 adulti; dagli animali e da persone sono stati raccolti 68 adulti e 5 ninfe appartenenti a diverse specie di Ixodidae. Sono state condotte analisi sull’abbondanza delle zecche nelle diverse aree di raccolta, in funzione del periodo di campionamento, della temperatura e dell’umidità relativa misurata a 5 cm dal suolo al momento del campionamento e della vegetazione. Su tutti gli individui adulti e su pool di ninfe e di larve, per un totale di 393 campioni, sono state condotte analisi di tipo molecolare per la ricerca di piroplasmi, Anaplasma phagocytophilum e Borrelia burgdorferi s.l. Attraverso la PCR e il sequenziamento, è emerso che il 7,6% dei campioni era positivo per piroplasmi, tra i quali è stata riscontrata anche la presenza delle specie zoonosiche Babesia EU1 e B. divergens. La real-time PCR eseguita solo sui campioni costituiti da ninfe e adulti ha evidenziato una prevalenza del 9,2% per A. phagocytophilum e del 21,6% per B. burgdorferi s.l. Su questi patogeni sono state quindi condotte analisi di tipo filogenetico. In alcuni campioni sono state riscontrate coinfezioni con combinazioni di due patogeni contemporaneamente.

Portal hypertension: a comparison between portal-venous pressure measurement and ARFI measurement of liver and spleen stiffness

PURPOSE. Portal pressure is measured invasively as Hepatic Venous Pressure Gradient (HVPG) in the angiography room. Liver stiffness measured by Fibroscan was shown to correlate with HVPG values below 12 mmHg. This is not surprising, since in cirrhosis the increase of portal pressure is not directly linked with liver fibrosis and consequently to liver stiffness. We hypothesized that, given the spleen’s privileged location upstream to the whole portal system, splenic stiffness could provide relevant information about portal pressure. Aim of the study was to assess the relationship between liver and spleen stiffness measured by Virtual Touch™ (ARFI) and HVPG in cirrhotic patients. METHODS. 40 consecutive patients (30 males, mean age 62y, mean BMI=26, mean Child-Pugh A6, mean platelet count=92.000/mmc, 19 HCV+, 7 with ascites) underwent to ARFI stiffness measurement (10 valid measurements in right liver lobe both surface and centre, left lobe and 20 in the spleen) and HPVG, blindly to each other. Median ARFI values of 10 samplings on every liver area and of 20 samplings on spleen were calculated. RESULTS. Stiffness could be easily measured in all patients with ARFI, resulting a mean of 2,61±0,76, 2,5±0,62 and 2,55±0,66 m/sec in the liver areas and 3.3±0,5 m/s in the spleen. Median HPVG was 14 mmHg (range 5-27); 28 patients showed values ≥10 mmHg. A positive significant correlation was found between spleen stiffness and HPVG values (r=0.744, p<0.001). No significant correlation was found between all liver stiffness and HVPG (p>0,05). AUROC was calculated to test spleen stiffness ability in discriminating patients with HVPG ≥10. AUROC = 0.911 was obtained, with sensitivity of 69% and specificity of 91% at a cut-off of 3.26 m/s. CONCLUSION. Spleen stiffness measurement with ARFI correlates with HVPG in patients with cirrhosis, with a potential of identifying patients with clinically significant portal hypertension.

New associations between native parasitoids and exotic insects introduced in Italy and in Europe

The introduction of exotic species is one of the most important threats to biodiversity.This phenomenon may cause economic and environmental damage. To prevent these invasions there are institutions like EPPO. Nevertheless, the introduction of exotic pests is an increasing issue, difficult to control. Classic biological control, based on importation of natural enemies from the country of origin, has been successfully used for over 120 years, but it has also raised some criticism. My research work has focused on the study of the new associations occurring between indigenous parasitoids and three exotic pests introduced in Italy and Europe. The three target insects considered were: Cacyreus marshalli Butler (Lepidoptera: Lycaenidae), a pest of Geranium plants; Dryocosmus kuriphilus Yasumatsu (Hymenoptera: Cynipidae), a plague of Castanea sp. and Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae). This ladybug has been introduced as a biological control agent, but since some years it considered as an invasive species. For C. marshalli I performed laboratory tests on acceptance and suitability of immature stages of this butterfly by Exorista larvarum (Diptera: Tachinidae) and Brachymeria tibialis (Hymenoptera: Chalcidicae). The experiments showed that these two parasitoids could be used to contain this pest. For D. kuriphilus I performed field samplings in an infested chestnut area, the samples were maintained in rearing chamber until gall wasp or parasitoids emergence. In the 3-year research many parasitoids of gall wasps were found; one of these, Torymus flavipes (Walker), was found in large number. For H. axyridis the research work included a first phase of field sampling, during which I searched indigenous parasitoids which had adapted to this new host; the only species found was Dinocampus coccinellae (Schrank) (Hymenoptera: Braconidae). Laboratory tests were performed on the wasp rearing, biology and capacity to contain H. axyridis.

Relationships between plant diversity and environmental heterogeneity in rupicolous grasslands on gypsum. The case study of Alysso-Sedion albi (Habitat 6110).

Plant communities on weathered rock and outcrops are characterized by high values in species richness (Dengler 2006) and often persist on small and fragmented surfaces. Yet very few studies have examined the relationships between heterogeneity and plant diversity at small scales, in particular in poor-nutrient and low productive environment (Shmida and Wilson 1985, Lundholm 2003). In order to assess these relationships both in space and time in relationship, two different approaches were employed in the present study, in two gypsum outcrops of Northern Apennine. Diachronic and synchronic samplings from April 2012 to March 2013 were performed. A 50x50 cm plot was used in both samplings such as the sampling unit base. The diachronic survey aims to investigate seasonal patterning of plant diversity by the use of images analysis techniques integrated with field data and considering also seasonal climatic trend, the substrate quality and its variation in time. The purpose of the further, synchronic sampling was to describe plant diversity pattern as a function of the environmental heterogeneity meaning in substrate typologies, soil depth and topographic features. Results showed that responses of diversity pattern depend both on the resources availability, environmental heterogeneity and the manner in which the different taxonomic group access to them during the year. Species richness and Shannon diversity were positively affected by increasing in substrate heterogeneity. Furthermore a good turnover in seasonal species occurrence was detected. This vegetation may be described by the coexistence of three groups of species which created a gradient from early colonization stages, characterized by greater slope and predominance of bare rock, gradually to situation of more developed soil.

Comparison of gingival crevicular fluid sampling methods in patients with severe chronic periodontitis

The analysis of samplings from periodontal pockets is important in the diagnosis and therapy of periodontitis. In this study, three different sampling techniques were compared to determine whether one method yielded samples suitable for the reproducible and simultaneous determination of bacterial load, cytokines, neutrophil elastase, and arginine-specific gingipains (Rgps). Rgps are an important virulence factor of Porphyromonas gingivalis, the exact concentration of which in gingival crevicular fluid (GCF) has not been quantified.

Vertical transmission of a fluoroquinolone-resistant Escherichia coli within an integrated broiler operation

The epidemiology of an enrofloxacin-resistant Escherichia coli clone was investigated during two separate outbreaks of colibacillosis in the Danish broiler production. In total five flocks were reported affected by the outbreaks. Recorded first-week mortalities were in the range of 1.7-12.7%. The clone was first isolated from dead broilers and subsequently demonstrated in samples from associated hatchers and the parent flock with its embryonated eggs, suggesting a vertical transmission from the parents. The second outbreak involved two broiler flocks unrelated to the affected flocks from the first outbreak. However, the clone could not be demonstrated in the associated parent flock. Furthermore, samplings from grand-parent flocks were negative for the outbreak clone. The clonality was evaluated by plasmid profiling and pulsed-field gel electrophoresis. None of the recognized virulence factors were demonstrated in the outbreak clone by microarray and PCR assay. The molecular background for the fluoroquinolone-resistance was investigated and point mutations in gyrA and parC leading to amino-acid substitutions in quinolone-resistance determining regions of GyrA and ParC were demonstrated. Vertical transmission of enrofloxacin-resistant E. coli from healthy parents resulting in high first-week mortality in the offspring illustrates the potential of the emergence and spreading of fluoroquinolone-resistant bacteria in animal husbandry, even though the use of fluoroquinolones is restricted.

Two-year outcome with Nobel Direct implants: a retrospective radiographic and microbiologic study in 10 patients

INTRODUCTION: The Nobel Direct implant (Nobel Biocare AB, Göteborg, Sweden) was developed to minimize marginal bone resorption and to result in "soft tissue integration" for an optimized aesthetic outcome. However, conflicting results have been presented in the literature. The aim of this present study was to evaluate the clinical and microbiologic outcomes of Nobel Direct implants. MATERIALS AND METHODS: Ten partially edentulous subjects without evidence of active periodontitis (mean age 55 years) received 12 Nobel Direct implants. Implants were loaded with single crowns after a healing period of 3 to 6 months. Treatment outcomes were assessed at month 24. Routine clinical assessments, intraoral radiographs, and microbiologic samplings were made. Histologic analysis of one failing implant and chemical spectroscopy around three unused implants was performed. Paired Wilcoxon signed-rank test was used for the evaluation of bone loss; otherwise, descriptive analysis was performed. RESULTS: Implants were functionally loaded after 3 to 6 months. At 2 years, the mean bone loss of remaining implants was 2.0 mm (SD +/- 1.1 mm; range: 0.0-3.4 mm). Three out of 12 implants with an early mean bone loss >3 mm were lost. The surviving implants showed increasing bone loss between 6 and 24 months (p = .028). Only 3 out of the 12 implants were considered successful and showed bone loss of <1.7 mm after 2 years. High rates of pathogens, including Aggregatibacter actinomycetemcomitans, Fusobacterium spp., Porphyromonas gingivalis, Pseudomonas aeruginosa, and Tanerella forsythia, were found. Chemical spectroscopy revealed, despite the normal signals from Ti, O, and C, also peaks of P, F, S, N, and Ca. A normal histologic image of osseointegration was observed in the apical part of the retrieved implant. CONCLUSION: Radiographic evidence and 25% implant failures are indications of a low success rate. High counts and prevalence of significant pathogens were found at surviving implants. Although extensive bone loss had occurred in the coronal part, the apical portion of the implant showed some bone to implant integration.