Feeding entrainment of locomotor activity and clock gene expression in Senegalese sole, Solea senegalensis

The present study was conducted to investigate the influence of restricted food access on Solea senegalensis behaviour and daily expression of clock genes in central (diencephalon and optic tectum) and pheripheral (liver) tissues. The Senegalese sole is a marine teleost fish belonging to the Class of Actinopterygii, Order Pleuronectiformes and Family Soleidae. Its geographical distribution in the Mediterranean sea is fairly broad, covering the south and east of the Iberian Peninsula, the North of Africa and Middle East until the coast of Turkey. From a commercial perspective Solea senegalensis has acquired in recent years, a key role in aquacolture industry of the Iberian Peninsula. The Senegalese sole is also acquiring an important relevance in chronobiological studies as the number of published works focused on the sole circadian system has increased in the last few years. The molecular mechanisms underlying sole circadian rhythms has also been explored recently, both in adults and developing sole. Moreover, the consideration of the Pleuronectiformes Order as one of the most evolved teleost groups make the Senegalese sole a species of high interest under a comparative and phylogenetic point of view. All these facts have reinforced the election of Senegalese sole as model species for the present study. The animals were kept under 12L:12D photoperiod conditions and divided into three experimental groups depending on the feeding time: fed at midlight (ML), middark (MD) or random (RND) times. Throughout the experiment, the existence of a daily activity rhythm and it synchronization to the light-dark and feeding cycles was checked. To this end locomotor activity was registred by means of two infrared photocells placed in pvc tube 10 cm below the water surface (upper photocell) and the other one was located 10 cm above the bottom of the tank (bottom photocell). The photocell were connected to a computer so that every time a fish interrupted the infrared light beam, it produced an output signal that was recorded. The number of light beam interruptions was stored every 10 minutes by specialized software for data acquisition.

Ciclo riproduttivo annuale del corallo solitario Caryophyllia inornata (Scleractinia, Caryophylliidae) caratterizzato da un inusuale modello di embriogenesi

La varietà dei processi riproduttivi dei coralli riflette la loro straordinaria abilità rigenerativa. Le scleractinie sono un noto esempio di animali clonali, che possono esibire una strategia mista di riproduzione sessuale e asessuale per mantenere la loro popolazione. Questo studio descrive per la prima volta il ciclo riproduttivo annuale e l’embriogenesi del corallo temperato Caryophyllia inornata. I campioni sono stati raccolti mediante immersioni subacquee dal relitto di un aereo, a largo delle coste dell’Isola d’Elba. Sono state eseguite analisi citometriche per definire lo sviluppo annuale delle cellule germinali e l’embriogenesi. Caryophyllia inornata è risultata gonocorica con una sex ratio di 1:3.3 a favore dei maschi. I polipi erano sessualmente maturi a 7 mm di lunghezza. Lo sviluppo degli embrioni è stato osservato in maschi, femmine ed individui sessualmente inattivi. Gli spermiari richiedevano circa 10 mesi di maturazione, mentre l’ovogenesi sembrava essere più rapida. I polipi femmina sono stati trovati solo da Febbraio a Luglio. Il tasso di sviluppo gonadico aumentava sensibilmente da Marzo a Maggio e la fecondazione avveniva tra Maggio e Luglio, quando le cellule germinali all’interno dei polipi iniziavano a scomparire. La gametogenesi ha mostrato una forte influenza stagionale mentre gli embrioni sono stati trovati durante tutto l’anno nei maschi e negli individui sessualmente inattivi, non presentando un andamento ben definito. Questo modello di embriogenesi è poco comune ed è stato osservato solo in quattro scleractinie tropicali (Pocillopora damicornis, Tubastrea Diaphana, T. coccinea e Oulastrea crispata) in cui avviene una produzione interna di embrioni per via agamica. La natura precisa di questo processo è ancora sconosciuta.

Studio dell'effetto dell'acidificazione del mare sulla riproduzione del corallo Astroides calycularis (Scleractinia, Dendrophylliidae) lungo un gradiente naturale di CO2

E’ previsto che l’emissione in atmosfera di anidride carbonica (CO2) da parte dell’uomo e la conseguente acidificazione delle acque dei mari provocherà effetti negativi di vasta portata sui processi biologici di diversi ecosistemi marini. L’acidificazione del mare può provocare effetti negativi su tutti gli stadi del ciclo vitale degli organismi marini, influenzandone la fisiologia, morfologia, comportamento ed alcune fasi della riproduzione e del reclutamento. Esistono poche informazioni riguardo gli effetti dell’acidificazione sulla riproduzione sessuale dei coralli. Questo studio ha lo scopo di quantificare la capacità riproduttiva di Astroides calycularis (scleractinia coloniale mediterranea azooxantellata) in colonie trapiantate lungo un gradiente naturale di pH, generato dalle emissioni di CO2 del cratere vulcanico sottomarino di Panarea (Isole Eolie). Le colonie sono state trapiantate in 4 siti, dal centro del cratere, dove si riscontra la condizione più acida (pH 7.40), alla periferia, caratterizzata da condizioni di pH normali (pH 8.07). La popolazione di A. calycularis di Panarea ha mostrato una sessualità mista a livello coloniale (polipi gonocorici in colonie ermafrodite), in contrapposizione alla condizione gonocorica riscontrata nella stessa specie studiata a Palinuro e a Punta de la Mona (Spagna). I risultati preliminari sembrerebbero mostrare un effetto negativo dell’acidificazione sullo sviluppo gonadico femminile della specie. L’ipotesi è che esso sia dovuto ad una riallocazione delle energie sui processi di calcificazione a danno dell’ovogenesi. La spermatogenesi risentirebbe meno degli effetti negativi dell’acidificazione nelle fasi di sviluppo interne all’ individuo, in quanto meno dispendiose da un punto di vista energetico rispetto alle fasi di ovogenesi. Gli spermatozoi potrebbero invece essere influenzati negativamente dall’ ambiente più acido una volta entrati in contatto con esso, con conseguenze sulla riuscita della fecondazione. Questo studio necessita di ulteriori dati e verrà completato con le analisi istologiche in corso.

Use of isotopes for studying nitrogen processes in a wetland within the Venice Lagoon watershed

The present study is based on the use of isotopes for evaluating the efficiency of nutrients removal of a wetland, in particular nitrogen and nitrates, also between the different habitats present in the wetland. Nutrients like nitrogen and phosphorus, normally distributed as fertilizers, are among the principal causes of diffuse pollution. This is particularly important in the Adriatic Sea, which is frequently subjected to eutrophication phenomena. So it is very crucial requalification of wetland, in which there are naturally depurative processes such as denitrification and plant uptake, which allow the reduction of pollutant loads that flow in water bodies. In this study nutrient reduction is analyzed in the wetland of the Comuna drain, which waters flow in the Venice lagoon. Chemical and isotopical analyses were performed on samples of water, vegetation, soil and sediments taken in the wetlands of the Comuna drain in four different periods of the year and on data of nitrogen and phosphorus concentration obtained by the LASA of the University of Padova. Values of total nitrogen and nitrates were obtained in order to evaluate the reduction within the different systems of the wetland. Instead, the isotopic values of nitrogen and carbon were used to evaluate which process influence more nitrogen reduction and to understand the origin of the nutrient, if it is from fertilizers, waste water or sewage. To conclude, the most important process in the wetland of the Comuna drain is plant uptake, in facts the bigger percentage of nitrogen reduction was in the period of vegetative growth. So it is important the study of isotopes in plant tissues and water residence time, whose increase would allow a greater reduction of nutrients.

Genetic structuring of Eunicella singularis populations along a bathymetric gradient

Phenotypic plasticity refers to the ability of an organism to express different morphologies depending on the abiotic and biotic environment. Depth integrating many variables (e.g. temperature, light and hydrodynamics), may affect population structure and dynamics of the populations, as well as connectivity patterns and genetic diversity. Eunicella singularis is a Mediterranean arborescent gorgonian who plays an important rule as engineer species providing biomass and complexity to coralligenous habitats. It has a wide bathymetric distribution ranging from shallow rocky bottoms to deep sublittoral reefs. The species shows two depth-related morphotypes which taxonomic status is not yet clarified. The aim of the study is to analyses genetic variability and/or structuring along a vertical gradient to test the presence of the two morphotypes. Furthermore, a preliminary analyses of the phylogenetic relationship among species of the genus Eunicella has been done. Six populations of Eunicella singularis were sampled from 10 to 60 m depth in Cap de Creus and individuals belonging to Eunicella cavolinii, E. verrucosa, E. racemosa and E. stricta aphyta were collected. The genetic analyses were carried out using five microsatellite loci and ITS-1 sequence polymorphism. The results showed a reduction of genetic variability along the vertical gradient. A threshold in connectivity was observed across 30 - 40 m depth, confirming the presence of two different Eunicella singularis morphotypes. The two morphological forms could be due to phenotypic plasticity, which allowed populations to suit different environmental conditions, or to a break in gene flow that determined the isolation of the two populations and an accumulation of genetic differences. The molecular markers used were not able to clarify the phylogenetic relationship among Eunicella species and the systematic position of the two morphotypes, conversely they risen the question on the existence of single species of Mediterranean Eunicella.

Modelling growth dynamics and toxin production in Ostreopsis cf. ovata

The benthic dinoflagellate O. ovata represents a serious threat for human health and for the ecology of its blooming areas: thanks to its toxicity this microalga has been responsible for several cases of human intoxication and mass mortalities of benthic invertebrates. Although the large number of studies on this dinoflagellate, the mechanisms underpinning O. ovata growth and toxin production are still far to be fully understood. In this work we have enriched the dataset on this species by carrying out a new experiment on an Adriatic O. cf. ovata strain. Data from this experiment (named Beta) and from another comparable experiment previously conducted on the same strain (named Alpha), revealed some interesting aspects of this dinoflagellate: it is able to grow also in a condition of strong intracellular nutrient deficiency (C:P molar ratio > 400; C:N > 25), reaching extremely low values of chlorophyll-a to carbon ratio (0.0004). Was also found a significant inverse relationships (r > -0.7) between cellular toxin to carbon and cellular nutrient to carbon ratios of experiment Alpha. In the light of these result, we hypothesized that in O. cf. ovata nutrient-stress conditions (intended as intracellular nutrient deficiency) can cause: i) an increase in toxin production; ii) a strong decrease in chlorophyll-a synthesis; iii) a lowering of metabolism associated with the formation of a sort of resting stage. We then used a modelling approach to test and critically evaluate these hypotheses in a mechanistic way: newly developed formulation describing toxin production and fate, and ad hoc changes in the already existent formulations describing chlorophyll synthesis, rest respiration, and mortality, have been incorporated in a simplified version of the European Regional Seas Ecosystem Model (ERSEM), together with a new ad hoc parameterization. The adapted model was able to accurately reproduce many of the trends observed in the Alpha experiment, allowing us to support our hypotheses. Instead the simulations of the experiment Beta were not fully satisfying in quantitative terms. We explained this gap with the presumed different physiological behaviors between the algae of the two experiments, due to the different pre-experimental periods of acclimation: the model was not able to reproduce acclimation processes in its simulations of the experiment Beta. Thus we attempt to simulate the acclimation of the algae to nutrient-stress conditions by manual intervention on some parameters of nutrient-stress thresholds, but we received conflicting results. Further studies are required to shed light on this interesting aspect. In this work we also improve the range of applicability of a state of the art marine biogeochemical model (ERSEM) by implementing in it an ecological relevant process such as the production of toxic compounds.

Diversity of microbial communities at shallow-hydrothermal vents off Dominica, Lesser Antilles

Hydrothermal vents are often compared to desert oases, because of the presence of highly diverse and abundant biotic communities inhabiting these extreme environments. Nevertheless, the microbial communities associated with shallow-hydrothermal systems have been poorly studied. Hydrothermal activity at Dominica Island is quite well known under the geological and geochemical aspects, but no previous information existed about the microbial communities associated to this area. This thesis is therefore targeting the microbiology of hydrothermal sediments combining geochemical and molecular biological investigations, focusing on differences between hydrothermal vents and background (i.e. control) areas, and between hydrothermal sites. It was also intended to assess relationship between geochemical parameters and microbial diversity at the two hydrothermally impacted sites. Two shallow-sea hydrothermal vents located south-west off Dominica Island (Lesser Antilles) have been investigated in this study: Champagne Hot Springs and Soufriere Bay offshore vent. During this study, sediments for geochemical and molecular analyses were collected every 2 cm from the two impacted areas and from two control sites not associated with hydrothermal activity; in situ temperatures measurements were also taken every 5 cm deep in the sediment for all the sites. A geochemical characterization of the sediment porewater was performed through the analysis of several elements’ concentrations (i.e. H2S, Cl-, Br-, SO42-, Fe2+, Na+, K+, B+, Si+). Microbial communities at the different sites were studied by Automated Intergenic Spacer Analysis (ARISA). Inspection of the operational taxonomic units (OTUs) distribution was performed, as well as statistical analyses for communities’ structure and composition differences, and for changes of β-diversity along with sediment geochemistry. Data suggested that mixing between hydrothermal fluids and seawater results in distinct different environmental gradients and potential ecological niches between the two investigated hydrothermal vents, reflecting a difference in microbial community structures between them.

Genetic and morphological variation and differentiation of Raja clavata populations in the European seas: marker development and preliminary data

In this study the population structure and connectivity of the Mediterranean and Atlantic Raja clavata (L., 1758) were investigated by analyzing the genetic variation of six population samples (N = 144) at seven nuclear microsatellite loci. The genetic dataset was generated by selecting population samples available in the tissue databases of the GenoDREAM laboratory (University of Bologna) and of the Department of Life Sciences and Environment (University of Cagliari), all collected during past scientific surveys (MEDITS, GRUND) from different geographical locations in the Mediterranean basin and North-east Atlantic sea, as North Sea, Sardinian coasts, Tuscany coasts and Cyprus Island. This thesis deals with to estimate the genetic diversity and differentiation among 6 geographical samples, in particular, to assess the presence of any barrier (geographic, hydrogeological or biological) to gene flow evaluating both the genetic diversity (nucleotide diversity, observed and expected heterozygosity, Hardy- Weinberg equilibrium analysis) and population differentiation (Fst estimates, population structure analysis). In addition to molecular analysis, quantitative representation and statistical analysis of morphological individuals shape are performed using geometric morphometrics methods and statistical tests. Geometric coordinates call landmarks are fixed in 158 individuals belonging to two population samples of Raja clavata and in population samples of closely related species, Raja straeleni (cryptic sibling) and Raja asterias, to assess significant morphological differences at multiple taxonomic levels. The results obtained from the analysis of the microsatellite dataset suggested a geographic and genetic separation between populations from Central-Western and Eastern Mediterranean basins. Furthermore, the analysis also showed that there was no separation between geographic samples from North Atlantic Ocean and central-Western Mediterranean, grouping them to a panmictic population. The Landmark-based geometric morphometry method results showed significant differences of body shape able to discriminate taxa at tested levels (from species to populations).

Identification of stock units of Mullus barbatus (Linnaeus, 1758) in the Italian seas: development of genomic 2b-RAD markers and biological variation analysis.

This thesis is settled within the STOCKMAPPING project, which represents one of the studies that were developed in the framework of RITMARE Flagship project. The main goals of STOCKMAPPING were the creation of a genomic mapping for stocks of demersal target species and the assembling of a database of population genomic, in order to identify stocks and stocks boundaries. The thesis focuses on three main objectives representing the core for the initial assessment of the methodologies and structure that would be applied to the entire STOCKMAPPING project: individuation of an analytical design to identify and locate stocks and stocks boundaries of Mullus barbatus, application of a multidisciplinary approach to validate biological methods and an initial assessment and improvement for the genotyping by sequencing technique utilized (2b-RAD). The first step is the individuation of an analytical design that has to take in to account the biological characteristics of red mullet and being representative for STOCKMAPPING commitments. In this framework a reduction and selection steps was needed due to budget reduction. Sampling areas were ranked according the individuation of four priorities. To guarantee a multidisciplinary approach the biological data associated to the collected samples were used to investigate differences between sampling areas and GSAs. Genomic techniques were applied to red mullet for the first time so an initial assessment of molecular protocols for DNA extraction and 2b-RAD processing were needed. At the end 192 good quality DNAs have been extracted and eight samples have been processed with 2b-RAD. Utilizing the software Stacks for sequences analyses a great number of SNPs markers among the eight samples have been identified. Several tests have been performed changing the main parameter of the Stacks pipeline in order to identify the most explicative and functional sets of parameters.

Stock boundaries of Parapenaeus longirostris (Lucas, 1846) populations in the Italian seas: Development of genomic 2b-RAD markers and biological variation analysis.

This thesis is developed in the contest of Ritmare project WP1, which main objective is the development of a sustainable fishery through the identification of populations boundaries in commercially important species in Italian Seas. Three main objectives are discussed in order to help reach the main purpose of identification of stock boundaries in Parapenaeus longirostris: 1 -Development of a representative sampling design for Italian seas; 2 -Evaluation of 2b-RAD protocol; 3 -Investigation of populations through biological data analysis. First of all we defined and accomplished a sampling design which properly represents all Italian seas. Then we used information and data about nursery areas distribution, abundance of populations and importance of P. longirostris in local fishery, to develop an experimental design that prioritize the most important areas to maximize the results with actual project funds. We introduced for the first time the use of 2b-RAD on this species, a genotyping method based on sequencing the uniform fragments produced by type IIB restriction endonucleases. Thanks to this method we were able to move from genetics to the more complex genomics. In order to proceed with 2b-RAD we performed several tests to identify the best DNA extraction kit and protocol and finally we were able to extract 192 high quality DNA extracts ready to be processed. We tested 2b-RAD with five samples and after high-throughput sequencing of libraries we used the software “Stacks” to analyze the sequences. We obtained positive results identifying a great number of SNP markers among the five samples. To guarantee a multidisciplinary approach we used the biological data associated to the collected samples to investigate differences between geographical samples. Such approach assures continuity with other project, for instance STOCKMED, which utilize a combination of molecular and biological analysis as well.

Stato tossicologico di esemplari di foca monaca (Monachus monachus, Hermann 1779) della Grecia, specie "critically endangered" del Mar Mediterraneo.

La foca monaca (Monachus monachus, Hermann 1779) è il pinnipede più a rischio d’estinzione del mondo e pochi sono gli studi che hanno indagato l’impatto antropico dato dagli inquinanti chimici su questa specie. Questo studio si propone di analizzare la pressione che elementi in traccia, organoclorurati (OC) e, per la prima volta, idrocarburi policiclici aromatici (IPA) hanno sulla specie, studiando una della popolazioni più numerose presenti nel Mar Mediterraneo, ossia quella greca. Sono stati utilizzati campioni di 59 esemplari deceduti provenienti da diverse colonie residenti in Grecia, di ambedue i sessi, appartenenti a diverse classi d’età e riferibili ad un arco temporali di quasi 20 anni (1994-2013). Per i contaminanti organici (PCB, DDT ed IPA), caratterizzati da un comportamento lipofilico, sono stati utilizzati esclusivamente campioni di tessuto adiposo, mentre per l’analisi di elementi in traccia (essenziali: Fe, Co, Cu, Se, Zn, Mg; tossici: Cr, Ni, As, Cd, Pb, Hg) sono state impiegate diverse matrici, quali fegato, rene, muscolo, adipe, milza, polmone, cuore e pelo. I risultati evidenziano un’importante contaminazione dai OC (125.896,69±289.934,27 ng/g p.s.), in cui DDT e PCB sono presenti rispettivamente al 48,5% e 51,5%, mentre, tra i metalli considerati tossici (As, Cd, Pb, Hg, Cr, Ni), As e Hg sono gli elementi che destano maggiori preoccupazioni per la salute di Monachus monachus, presenti rispettivamente al 36,44% e 12,83%. IPA ed altri elementi in traccia analizzati invece non raggiungono di per sé concentrazioni di rischio per la specie, ma non si escludono possibili effetti sinergici tra i vari inquinanti. L’elaborazione del fingerprint tossicologico ha evidenziato un’eterogeneità nella pressione data dai contaminanti in studio, manifestando che alcune colonie di foca monaca in Grecia sono maggiormente soggette all’azione negativa di organoclorurati, idrocarburi policiclici aromatici o elementi in traccia tossici a seconda del caso.

Osservazioni in ambiente controllato del ciclo riproduttivo e delle prime fasi di accrescimento di Chiloscyllium punctatum (Muller & Henle 1838)

La presente tesi tratta delle attività riproduttive e delle prime fasi di crescita dello squalo Chiloscyllium punctatum. Da Dicembre 2013 a Giugno 2014, all’Acquario di Cattolica, sono state effettuate osservazioni su una coppia di adulti riproduttori e sui relativi neonati mantenuti in ambiente controllato. Il Chiloscyllium punctatum è una specie ovipara che rilascia uova di forma rettangolare, ha un ciclo riproduttivo relativamente rapido ed è molto sfruttata come specie ornamentale dagli acquari di tutto il mondo. Per ottenere una corretta gestione in ambiente controllato, la riproduzione ed estrapolare il maggior numero d’informazioni certe su gli stadi di vita, sulla dieta di divezzamento dei neonati dal sacco vitellino, sull’illuminazione più consona da adottare, sulle dimensioni medie di uova, nascituri e adulti, sulla crescita dei nascituri, sui tempi di sviluppo embrionale, sulla frequenza di deposizione delle uova e sui tempi di schiusa è stato necessario sviluppare un corretta procedura di gestione ordinaria. In 7 mesi, gli adulti di C. punctatum, in condizioni ambientali ottimali e costanti, hanno dato vita a 33 neonati. Questi sono stati sottoposti a misurazioni di lunghezza (LT, cm) e peso corporeo (PC, g) per valutare l’influenza dell’illuminamento e della dieta sulla crescita. Inoltre, sono stati confrontati dati sperimentali (e.g. tempistiche di sviluppo embrionale) con dati bibliografici (Haraush et al., 2007) che hanno evidenziato alcune similitudini e differenze.

Studio della presenza di capodogli (Physeter macrocephalus, Linnaeus 1758) in due aree del Mar Balear mediante metodi acustici passivi

I capodogli (Physeter macrocephalus) sono mammiferi marini con distribuzione cosmopolita; sono presenti anche nel Mar Mediterraneo, soprattutto nella zona occidentale del bacino. Per individuare la presenza di questi animali possono essere utilizzate differenti modalità. Questo studio è stato condotto tramite l’uso di metodi acustici passivi, in quanto i capodogli (P. macrocephalus) sono animali che emettono suoni. L’uso di metodi acustici ha alcuni vantaggi, infatti permette di individuare la presenza di un animale anche se si trova al di sotto della superficie dell’acqua, purché stia emettendo suoni. Le registrazioni possono essere effettuate indipendentemente dalle condizioni meteorologiche e per lunghi periodi di tempo, permettendo la continuità nella raccolta dei dati. Il lavoro di tesi è stato svolto nell’ambito di un Progetto di Ricerca de L’Oceanogràfic di Valencia. Le registrazioni sono state effettuate tramite lo strumento EAR (Ecological Acoustic Recorder) in due aree del Mar Balear, il Mar di Bamba e il Parco Nazionale di Cabrera, in differenti periodi, profondità e con diverse frequenze di campionamento (50 kHz e 5 kHz). I dati ottenuti sono stati analizzati tramite i programmi X-BAT (Extensible Bioacoustic Tool) e Adobe Audition 3.0 al fine di rilevare i pulsi emessi dai capodogli. Per entrambe le aree di studio i dati sono stati elaborati in modo da ottenere modelli giornalieri e stagionali delle ore positive e del numero di pulsi emessi. La presenza di P. macrocephalus è stata confermata in entrambe le aree di studio, e dal confronto delle analisi spettrali dei pulsi si può affermare che, nonostante le differenze, i pulsi registrati in entrambe le aree sono effettivamente emessi da capodogli. Il lavoro di Tesi permette di suggerire che per futuri studi sia applicata la frequenza di registrazione di 20 kHz utilizzando lo strumento SAMARUC, alla cui progettazione ha partecipato L’Oceanogràfic di Valencia.

Distribuzione dei neuroni che esprimono il peptide correlato al gene della calcitonina (CGRP) nel midollo spinale di Tursiope (Tursiops truncatus, Montagu, 1821)

Il peptide correlato al gene della calcitonina (CGRP) è una molecola presente nei neuroni del midollo spinale di diverse specie di Mammiferi, inclusi topi, ratti, conigli, cani, gatti, pecore, scimmie e uomo. Nonostante la distribuzione dei neuroni contenenti questo neuropeptide sia stata studiata in maniera dettagliata nel midollo spinale delle suddette specie, non sono disponibili, in letteratura, informazioni relative alla presenza di queste cellule nel midollo spinale dei Cetacei. Di conseguenza, è stata condotta la presente ricerca che ha avuto lo scopo di determinare, mediante metodiche di immunoistochimica, la distribuzione e la morfologia dei neuroni esprimenti il CGRP nel midollo spinale di tursiope (Tursiops truncatus). In questa specie, la distribuzione laminare (secondo Rexed) dei neuroni CGRP-immunoreattivi è assai simile a quella che si osserva nei Roditori, nei Carnivori e nei Primati; infatti, i corpi cellulari immunopositivi sono localizzati soprattutto in corrispondenza dell’apice del corno dorsale (lamine I e II) e nel corno ventrale (lamine VIII e IX). La distribuzione e la morfologia dei neuroni esprimenti CGRP nel midollo spinale di tursiope suggeriscono come tale neuropeptide possa essere coinvolto nella trasmissione delle informazioni sia sensitive (somatiche e viscerali) che motorie. I neuroni CGRP-immunoreattivi localizzati nelle lamine I e II del midollo spinale di tursiope, come dimostrato in altre specie, potrebbero agire da interneuroni modulando le informazioni nocicettive che dai gangli spinali vengono trasmesse al midollo spinale. Nelle lamine I e II sono presenti anche numerosi processi immunopositivi che, oltre ad appartenere a neuroni locali, derivano, molto probabilmente, dai ai neuroni pseudounipolari dei gangli spinali. In accordo con quanto appena affermato, è opportuno sottolineare come le fibre afferenti primarie provenienti dai gangli spinali utilizzino il CGRP per la trasmissione delle informazioni dolorifiche. La presenza di CGRP nei neuroni della lamina VIII, invece, indica come questo neuropeptide possa essere implicato nella trasmissione di segnali di natura motoria, utilizzando meccanismi presinaptici. Infine, la presenza di numerosi motoneuroni immunoreattivi per il CGRP nella lamina IX indicherebbe un’azione diretta svolta da questo neuropeptide nell’interazione tra motoneurone inferiore e muscolo scheletrico.

Evaluation of green algae deployment for organic pollutant removal in urban wastewater through ERE-CALUX potency assessment

The interest of the scientific community towards organic pollutants in freshwater streams is fairly recent. During the past 50 years, thousands of chemicals have been synthesized and released into the general environment. Nowadays their occurrence and effects on several organism, invertebrates, fish, birds, reptiles and also humans are well documented. Because of their action, some of these chemicals have been defined as Endocrine Disrupters Compounds (EDCs) and the public health implications of these EDCs have been the subject of scientific debate. Most interestingly, among those that were noticed to have some influence and effects on the endocrine system were the estrone, the 17β-estradiol, the 17α-estradiol, the estriol, the 17α-ethinylestradiol, the testosterone and the progesterone. This project focused its attention on the 17β-estradiol. Estradiol, or more precisely, 17β-estradiol (also commonly referred to as E2) is a human sex hormone. It belongs to the class of steroid hormones. In spite of the effort to remove these substances from the effluents, the actual wastewater treatment plants are not able to degrade or inactivate these organic compounds that are continually poured in the ecosystem. Through this work a new system for the wastewater treatment was tested, to assess the decrease of the estradiol in the water. It involved the action of Chlorella vulgaris, a fresh water green microalga belonging to the family of the Chlorellaceae. This microorganism was selected for its adaptability and for its photosynthetic efficiency. To detect the decrease of the target compound in the water a CALUX bioassay analysis was chosen. Three different experiments were carried on to pursue the aim of the project. By analysing their results several aspects emerged. It was assessed the presence of EDCs inside the water used to prepare the culture media. C. vulgaris, under controlled conditions, could be efficient for this purpose, although further researches are essential to deepen the knowledge of this complex phenomenon. Ultimately by assessing the toxicity of the effluent against C. vulgaris, it was clear that at determined concentrations, it could affect the normal growth rate of this microorganism.