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Within the framework of the European project LIFEOONAT/IT/7216 for conservation of environments of endangered bats, in vitro propagation of Dianthus balbisii Ser. ssp. liburnicus (Bartl.) Pign. (Caryophyllaceae) from wild plants in the Onferno Natural Reserve has been planned to restore foraging habitats. A satisfactory percentage of not contaminated explants has been achieved after double sterilisation of nodal segments. Shoot proliferation occurred on a MS based medium enriched with 0.5 mg L -1 BAP and 0.7% commercial agar (B&V, Parma, Italy, S1000). After a short in vitro phase on a medium enriched with 1 mg L -1 IBA for root induction, acclimatisation in the greenhouse was successful. Plants have already been transplanted to outdoor flowerbeds in the Reserve. Different characteristics shown during proliferation and after acclimatisation by clones, each derived from single initial shoot tip, are still under observation.

Environmental pollution represents one of the main threats for marine ecosystems conservation. Top predators are particularly exposed to contaminants; due to their anatomical and physiological characteristics, cetaceans can reach very high pollutants tissue body burdens (i.e. organochlorine and heavy metals). These marine mammals are thus considered as good indicators of marine environmental pollutants and among them heavy metals are one of the major contaminants classes. Present research reports about the quantification of heavy metals in tissues of stranded bottlenose dolphins (Tursiops truncatus) from Northern Adriatic Sea and presents some speculation on possible subtle adverse effects exerted by these levels on the general health of the animals. Stranded dolphins were collected along Northern Adriatic Sea coasts during a 5 years period and underwent necropsy for the determination of the causes of death. During examination, organs were sampled and stored at -20°C until analysis, which was performed with an Inductively Coupled Plasma-Atomic Emission Spectroscopy (ICP-AES) technique. Quantified metals were Cr, Ni, Cu, Zn, Hg, As, Se, Cd and Pb and data are reported as mg/kg (ppm) on a wet weight basis. When possible, gender and age of the animals were recorded. All metals were at detectable levels, presenting a huge variation among animals and tissues. Interestingly, some of the most common contaminants for marine environment were detected at low levels, often below limit of quantification, also in accumulation target organs like liver and kidney. Mean heavy metals detected were to be considered as low, but high enough to exert a potential immunotoxic effect on animals, as the levels detected are comparable to those which proved to be active in vitro on lymphocytic cytotoxicity, apoptosis, lymphocytic proliferation and phagocytic activity, as confirmed by some anatomo-pathological observations of lymph node swelling. Some speculation on potential reproductive effect of detected heavy metals levels are also proposed, even if no clear evidence of altered reproductive activity was observed. From obtained data there seems to be a reduced exposure to heavy metals, even thought a higher susceptibility to infectious diseases can be considered for studied animals.

Leatherback turtles Dermochelys coriacea are regularly reported in the Mediterranean Sea but rarely reach the northern Adriatic Sea. In the summer of 2009, a well-preserved carcass of an adult female of this species was found dead along the coast of Lido di Venezia. A complete necropsy was carried out along with tissue trace element levels evaluation. The results of the postmortem studies revealed an acute severe bacterial gastroenteritis caused by Photobacterium damselae subsp. piscicida, an opportunistic agent that infected an apparently debilitated animal weakened by ingested plastic debris. High levels of heavy metals (Hg, Pb, Cd, As) found in liver and kidneys might have contributed to the animal’s demise. These findings support previous indications that marine debris is one of the major threats to marine animals, particularly for critically endangered species such as the leatherback turtle.

This study was conducted to determine heavy metals concentrations in tissues of 94 birds belonging to different species from coastal areas of Naples and Salerno (Southern Italy) in order to provide baseline data concerning trace element levels in wild birds living in or close to an area characterized by high anthropogenic impact. Additional aim of this study was to verify if diet influenced metal accumulation, so birds were classified as birds of prey, fish eating birds and insectivorous birds. Kidney was the primary organ for Cd accumulation in all groups considered, Pb was accumulated preferentially in bone, whereas Hg showed high values in liver of fish eating birds. Zn showed the highest mean concentrations, while Cu levels were one order of magnitude less. The variance analysis with respect to feeding habits disclosed only a lower accumulation of Hg in insectivores with respect to the other groups. For all metals, the concentrations measured in tissues should be considered indicative of chronic exposure to low, “background” environmental levels and/or to the presence of low bioavailable metals in the environment. In addition, metal levels were not of toxicological concern, being always well below the toxic thresholds defined for each metal.

The Mediterranean toothcarp (Aphanius fasciatus) can be used to biologically control mosquito larvae. The reproductive performance of 234 Mediterranean toothcarp (180 females and 54 males) was investigated. Reproductive factors were determined in fish kept in fresh or saline water (males and females together), and in fish kept separately by sex in saline water and hormonallytreated with carp pituitary extract (CPE). In the hormonally-treated group, the combined effect of temperature, photoperiod, and hormonal treatment induced the best ovary maturation and larvae production rates. The rate of reproduction was very high (average eggs/female: 5.9-10.5) compared to natural reproduction in the wild (average eggs/female: 2-4). Additionally, the hatching rate was 97-100%. The results of this study show that reproduction of Mediterranean toothcarp can be controlled in an artificial environment: the lack of mortality in adults during acclimation and conditioning in fresh and marine waters indicates good domestication and plasticity in reproductive parameters. The IJA appears exclusively as a peer-reviewed on-line open-access journal at http://www.siamb.org.il. Only registered persons are able to access full papers. To register free of charge, complete and return our registration form.

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The north western coast of Adriatic sea is characterized by high level of urbanization, an intensive touristic season, overfishing and high ship traffic. Such a strong fishing effort is severely impacting ecosystems and it is the major direct (bycatch) or indirect (food competition) threat for cetaceans. In the area the most common marine mammal is the bottlenose dolphin Tursiops truncatus. A series of random trips with small boats were carried out in April and May 2010 in the area between the river Po delta and river Reno estuary, for 268 hours and 1079 nautical miles of transects. Eleven percent of observation time was spent in areas between 1 to 5 miles from the shore, 38% from 5,1 to 10, 6% from 10,1 to 15, 29% from 15,1 to 25 and 16% from 25,1 to 35 miles of distance. Sightings were 2 in April, one with 1 adult specimen and one with 3 adults, and 4 in May, one with 3 bottlenose dolphins and three encounters with groups of 10, 15 and 35 specimens respectively. The distance from the coast was equally distributed between the distance ranges 1-5 miles, 10-20 miles and 20-30 miles. No correlation with meteorological and sea condition were found. This first results confirm the presence of a viable population of bottlenose dolphins in the area. Further observations will be dedicated to understand if there is one large group or independent sub-units along the whole 150 km coastal area and to start a photo identification activity that can help understanding movement, social structure and conservation necessities.

Il declino delle popolazioni di Anfibi è un’urgenza planetaria della conservazione. Le specie in pericolo o addirittura estinte stano divenendo un dato di fatto in tutti i continenti. Il contributo si volge a presentare una breve sintesi dei fattori concausanti questo fenomeno e stigmatizzare il ruolo soprattutto dei contaminanti quali fattori di deperimento diretto o fondamentali induttori negli squilibri con parassiti, infezioni e connessioni eco-sistematiche. Un altro ruolo fondamentale in molti ambiti è poi rappresentato dall’ingresso nei fragili ecosistemi acquatici di specie aliene predatrici che squilibrano i rapporti preda-predatore favorendo ulteriormente il crollo dei popolamenti. In questo quadro si presenta inoltre qui un progetto di ricerca relativo alla salute delle popolazioni di alcuni anfibi che la Provincia di Ravenna ha promosso recentemente e che è in carico al Corso di Laurea in Acquacoltura ed Igiene delle Produzioni Ittiche di Cesenatico della Facoltà di Medicina Veterinaria. Lo studio valuta presenze, comunità, struttura dei popolamenti e presenza di parassiti e contaminanti. Una particolare attenzione è naturalmente rivolta al fungo Batrachochytrium dendrobatidis così come ai fattori che ne potrebbero far sviluppare la virulenza. Ogni sito esaminato è caratterizzato in termini di struttura territoriale, caratteristiche dell’area umida e tasso di potenziale predazione da parte di uccelli e di Procambarus clarkii, astacide americano che si è diffuso con ingenti problemi in tutto il bacino padano e non solo. Lo studio parassitologico è focalizzato sulla presenza di organismi intestinali, cutanei ed ematici. I dati ottenuti saranno messi in relazione con la presenza di vari contaminanti (metalli e pesticidi) al fine di valutare se questi ultimi possono o meno influire sul grado di infestazione delle varie popolazioni. I dati complessivi confluiranno in un’analisi che speriamo possa dare indicazioni di piano per la realizzazione futura di azioni di salvaguardia.

Spinal ganglia (SG) contain the somata of primary afferent neurons that transmit sensory information from the periphery into the spinal cord. A subpopulation of ganglionic neurons containing neuronal nitric oxide synthase (nNOS), an enzyme known to generate nitric oxide, has been detected in a number of mammalian species. In this study, the presence of nNOS-immunoreactive (IR) neurons was examined in the bottlenose dolphin SG by immunohistochemical techniques. Double immunostaining was used to verify whether there was colocalization of nNOS with substance P (SP). In all SG, a subpopulation of small- and medium-sized neurons exhibited nNOS immunoreactivity. Data analysis revealed that the majority of nNOS-IR neurons expressed SP. On the contrary, few SP-IR neurons were also nNOS-positive The density of nNOS-immunoreactive and nNOS/SP-double immunopositive cells was relatively constant throughout the ganglia. The results of the present study suggest that nNOS-IR neurons may be involved in the afferent transmission of visceral and nociceptive information as well as in the control of the flow circulation in the retia mirabilia.

Heavy metals can be considered among major marine pollutants, presenting toxicological and accumulation problems in different environmental compartments. Even if they are present in all living organisms, their passage from one level to the other of the trophic chain is till now little or no studied. In present work we wanted to evaluate the transfer intensity of heavy metals (As, Cd, Co, Cr, Cu, Fe, Hg, Ni. Pb, Se) along a short, 4 steps trophic chain, composed by benthic and pelagic species, ending with bottlenose dolphin (Tursiops truncatus) as top predator. Samples (n= 30) from 3 species belonging to trophic level (TL) 1, 7 from TL 2, 5 from TL 3 and finally from tissues of stranded bottlenose dolphins were collected in Romagna area (Italy). Samples were microwave digested and analyzed with ICP-OES technique. Analytical results were used to calculate the biomagnification factor (BMF), as ratio between the upper level and the lower level specie concentration (BMF= Cn/Cn-1). When bottlenose tissue were considered, the BMF was calculated for each single tissue, to identify the “marker” tissue for each metal. Obtained results underline how contamination levels are to be considered as low for all of the species and all of the metals, with the only exception of Natica hebraea and Hinia reticulate, presenting very high levels of arsenic. Notable is the data concerning BMF calculation, that showed to be extremely variable not only depending on species, but also on different trophic level. Indeed, BMF for all toxic metals is lower than 1 in higher and lower levels, while it is always higher than 1 in intermediate level. This data is indicative of a strong concentration of toxicant in this trophic level. As concerns essential metals, BMF is always higher than 1, as expected considering their physiological role, thus indicating an active absorption and accumulation. Such an accumulation profile is to our knowledge not so common and presents interesting aspects concerning contaminants transfer, as it seems to underline, at least in the short food-chain we considered, a basic and “amplifying” role for intermediate predators in toxicant accumulation. It should anyway remembered that the contribution of each of the species considered to the contaminant burden of the species at upper level is influenced by the percent these species represent in total diet of their predators

Vitellogenin (VTG) is considered as a marker of endocrine disruption. A Western blot method for VTG quantification in Caretta caretta turtle plasma was developed using anti‐VTG antibody for Chelonia mydas. A screening of samples ( \documentclass{aastex} \usepackage{amsbsy} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{bm} \usepackage{mathrsfs} \usepackage{pifont} \usepackage{stmaryrd} \usepackage{textcomp} \usepackage{portland,xspace} \usepackage{amsmath,amsxtra} \usepackage[OT2,OT1]{fontenc} \newcommand\cyr{ \renewcommand\rmdefault{wncyr} \renewcommand\sfdefault{wncyss} \renewcommand\encodingdefault{OT2} \normalfont \selectfont} \DeclareTextFontCommand{\textcyr}{\cyr} \pagestyle{empty} \DeclareMathSizes{10}{9}{7}{6} \begin{document} \landscape $n=61$ \end{document} ) collected in the southern Mediterranean Sea around Lampedusa Island, Italy, was performed. The antibody showed a good cross‐reactivity with C. caretta VTG, suggesting a certain conservation of the core of the protein in dif...

E' stata valutata l’incidenza dell’età di riproduttori di trota fario sulla qualità delle uova e sull’entità di larve prodotte. Sono state analizzate le principali performance riproduttive di un gruppo di fattrici di taglia ≤ 37 cm (STG) e ≥ 45 cm (STV) stoccate presso un centro di ripopolamento dell’Appennino pesarese. I dati sono stati sottoposti ad analisi statistica. L’età delle femmine selezionate per la sperimentazione è risultata di 3-4+ nei soggetti STG e di 5-8+ nelle femmine STV. I risultati relativi alla riproduzione hanno evidenziato una migliore resa delle femmine giovani (STG) in termini di numero di uova emesse/100g di prodotto (1428 ±97 in STG, 719 ±58 in STV), di fecondità relativa (2777 ±410 in STG, 1305 ±400 in STV) e di percentuale di fecondazione (92,6 ±4,4 in STG, 79,4 ±7,3 in STV). Quanto rilevato infine ha determinato una maggiore produttività in termini di quantità di larve prodotte/kg peso vivo (2495 ±369 in STG, 974 ±298 in STV). Non sono emerse differenze significative tra i due gruppi in termini di numero di femmine che hanno deposto, di Indice Gonado-Somatico e di percentuale di schiusa. Le minori performance delle femmine più anziane evidenziano la necessità del rinnovo dello stock dei riproduttori.

Nell’ambito di un progetto di ampio respiro sull’ecologia e la riproduzione del Nono (Aphanius fasciatus) sono state indagate le performance riproduttive di uno stock proveniente da popolazioni della costa adriatica settentrionale. Oltre a rilevare l’andamento dell’indice gonadosomatico nelle diverse stagioni la riproduzione è stata indotta, cumulando gli effetti del condizionamento ecofisiologico e l’apporto di estratto pituitario di carpa, sia in acque dolci che salmastre. Temperatura, fotoperiodo e trattamento ormonale hanno permesso la maturazione ovarica e produzione di larve in tutte le femmine. Il tasso di schiusa è stato alto e il numero finale medio di larve per femmina si è attestato tra 3,33 e 7,43. L’allevamento di A. fasciatus in ambiente artificiale risulta essenziale per la produzione massiva utile a poter utilizzare questa specie autoctona e in pericolo per il controllo biologico delle larve di zanzara.

This work reports about the levels of heavy metals in female leatherback turtles and in their eggs and evaluates any potential correlation between blood heavy metals levels and thyroid hormones concentration. No correlation was found between mothers and eggs metals concentration nor between hematic metals and hormonal levels. Exposure levels in leatherbacks seems to be low, and there seems to be no active transfer of metals to the eggs, with the only exception of arsenic and chromium. Thus there seems to be no risk for both adult females and eggs from the exposure to environmental levels of heavy metals. Nevertheless, a certain concern arises for As and Cr, which should be further investigated for their toxic action on embryos.

I metalli pesanti rappresentano tra i principali contaminanti dell’ambiente marino, sia dal punto di vista tossicologico che di concentrazioni nei vari compartimenti ambientali. Sebbene presenti in tutti gli organismi viventi, il loro passaggio da un livello all’altro delle catene trofiche marine è ad oggi poco studiato ed approfondito. Nel presente lavoro si è voluto valutare l’intensità del trasferimento di metalli pesanti (As, Cd, Co, Cr, Cu, Fe, Hg, Ni. Pb, Se) lungo una catena trofica breve, composta da soli 4 livelli sia bentonici che pelagici, che partono da consumatori primari per arrivare ai tursiopi (Tursiops truncatus). A tal fine, sono stati prelevati campioni (n= 30) di 3 specie appartenenti al livello trofico 1, 7 specie del livello trofico 2, 5 di quello 3 ed infine tessuti diversi da esemplari di tursiope spiaggiati lungo le coste della Romagna. I campioni sono stati opportunamente mineralizzati ed analizzati con tecnica ICP-OES. I risultati analitici sono poi stati utilizzati per il calcolo del fattore di biomagnificazione (BMF), inteso come rapporto tra la concentrazione nella specie del livello superiore e quello del livello inferiore. I risultati ottenuti dalle analisi consentono innanzitutto di definire come i livelli di contaminazione osservati non siano da ritenersi elevati per nessuna delle specie considerate tranne che nel caso di Natica hebraea e di Hinia reticulata che presentano elevati livelli di arsenico. Dati interessanti vengono dall’analisi del BMF, che risulta estremamente variabile non solo in funzione delle specie, ma anche dei livelli trofici considerati. Infatti se il BMF è inferiore ad 1 nel caso dei livelli più alti e più bassi per tutti i metalli tossici, per il livello intermedio questo è sempre superiore a 1, indicando una forte concentrazione di contaminanti in questa fascia trofica. Per quanto riguarda gli elementi essenziali, come per altro immaginabile, il BMF è invece sempre superiore a 1, indicando un loro attivo assorbimento ed accumulo. Un simile profilo di accumulo è a nostra conoscenza non comune e presenta aspetti interessanti dal punto di vista del trasferimento dei contaminanti perché sembra delineare, per lo meno nella catena corta da noi considerata, un ruolo fondamentale ed “amplificante” dei predatori intermedi nell’accumulo degli inquinanti. Va per altro sottolineato come il contributo che ciascuna di queste specie fornisce al carico totale di metalli del livello superiore viene influenzato anche dalla percentuale che esse rappresentano nella dieta dei loro predatori.

This article reports the results of necropsy, parasitologic, microbiologic, histopathologic, immunohistochemical, indirect immunofluorescence, biomolecular, and serologic investigations on 8 striped dolphins ( Stenella coeruleoalba) found stranded from August to December 2007 on the Ligurian Sea coast of Italy. Severe, nonsuppurative meningoencephalitis was found in 4 animals, as characterized by prominent perivascular mononuclear cell cuffing and macrophage accumulations in neuropil. These lesions were associated with mild lymphocytic–plasmacytic infiltration of choroid plexuses in 1 dolphin. Toxoplasma gondii cysts and zoites, confirmed by immunohistochemical labeling, were scattered throughout the brain parenchyma of 2 of the 4 dolphins. No viral inclusions were seen in the brain of any animal. Other findings included severe bronchointerstitial pneumonia and pulmonary atelectasis, consolidation, and emphysema. Parasites were identified in a variety of organs, including lung ( Halocerchus lagenorhynchi). Microbiologic and serologic examinations for Brucella spp were negative on all 8 dolphins. The 4 animals with meningoencephalitis had serum antibodies against T gondii (titers ranging from 1:80 to 1:320) but not against morbillivirus. In contrast, the other 4 dolphins were seropositive for morbillivirus (with titers ranging from 1:10 to 1:40) but seronegative for T gondii. No morbillivirus antigen or nucleic acid was detected in the tissues of any dolphin. It is concluded that the severe lung and brain lesions were the cause of death and that T gondii was the likely etiologic agent of the cerebral lesions. Morbillivirus infection was not considered to have contributed to death of these animals.

Age determination based on the examination of hard anatomical parts is of fundamental importance in fisheries research. The most common method used for shark age assessment is the investigation of growth rings in vertebrae. As considerable variability exists in the amount and pattern of calcification within and among taxonomic groups of elasmobranch fishes, it is necessary to adopt a species-specific approach. The aim of the study was to employ radiographic and histological approaches to determine the age of a specimen of Cetorhinus maximus caught close to Ancona (Italy). Macroscopic evaluation of vertebral centra of three different vertebral segment chosen showed some difference among the numbers of rings. X-radiographs showed from 8 to 10 rings in the different vertebral bodies selected and histology of vertebral centra permitted to enhance the visibility of growth bands and to count up to 10 vertebral growth rings confirming data obtained by macroscopic evaluation and X-radiographs. According to the data obtained and considering a growth rings pattern of two depositions per year in this species as reported in literature, the young female here checked seem to be between the 4th and 5th year.

La presente ricerca è volta alla quantificazione di composti organoclorurati nel plasma di esemplari di Caretta caretta del bacino del Mediterraneo, ricoverati presso il Centro Regionale Recupero Fauna Selvatica e Tartarughe Marine del Fondo Siciliano per la Natura di Comiso. I dati analitici indicano una esposizione a vari composti organo clorurati e PCBs a livelli sufficientemente alti da indurre una alterazione del sistema immunitario in molti dei soggetti studiati. I dati ottenuti da esemplari per i quali erano disponibili campioni in entrata e in uscita dal centro di recupero, sebbene scarsi numericamente, sembrano indicativi di una modificazione del profilo residuale indotta dal cambiamento di alimentazione durante la fase di stabulazione, a testimoniare l’importanza della dieta nell’accumulo di questi composti persistenti e la validità e sensibilità del sangue nel monitoraggio dell’esposizione a contaminanti ambientali.

Starting from UNEP report 1999 Italy is first producer for heavy metals (Pb, Cd, Cu, Zn) marine pollution, being responsible of 30% of total reelase of these compounds in Mediterranean Sea. Being it a closed sea, with a reduced hydric exchange, Mediterranean is particularly exposed to risks derived from chemical pollution. Knowing pollution degree of this sea by estimating contaminants concentrations in marine species placed at the top of food chains, including marine mammals and cetacean, is thus mandatory. Despite the high number of studies focusing on heavy metals in tissues of different cetacean species, little information is available concerning contaminants transfer along their trophic chains, thus defining which could be main diet components contributing to toxicant body burden for each of the metal considered. Present work focuses on the evaluation of heavy metals presence in tissues of bottlenose from Northern Adriatic Sea and in some components of its diet, trying to define which could be the contribution that each of them give to the body burden of predators.

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Il progetto posto in essere dal laboratorio di Ecotossicologia del Dipartimento di Sanità Pubblica Veterinaria e Patologia Animale di Bologna è finalizzato alla valutazione dei livelli di metalli pesanti in specie diverse ricoverate negli zoo. Grazie alla collaborazione col Parco Zoo di Falconara si è iniziato il monitoraggio su specie che afferiscono a taxa con alimentazione differenziata, dei quali sono note la storia e la condizione clinica. Per la fase iniziale del progetto si sono utilizzati soggetti deceduti per cause naturali o per malattia, in modo da poter analizzare vari organi interni degli stessi. In particolare sono stati analizzati due saimiri, un mara, due giraffe e una leonessa. I campioni di organi (rene, fegato, muscolo, polmone, milza, midollo) sono stati prelevati in fase autoptica e conservati a -20°C fino al momento delle analisi, eseguite con tecnica ICP-OES per la ricerca di piombo, arsenico, mercurio, cadmio, cobalto, cromo, rame, ferro, selenio, manganese e zinco. I risultati analitici sono stati poi messi in relazione con lo stato di salute degli animali e con la vicinanza alle strade di accesso allo zoo, per verificare un’eventuale influenza del traffico veicolare sul contenuto di metalli. I primi risultati sembrano indicare una ridotta contaminazione da metalli pesanti in tutti gli esemplari esaminati, a prescindere dalla loro alimentazione, dalla loro storia clinica e dalla vicinanza alle strade, ma il ridotto numero di animali disponibili non permette di giungere alla definizione di valori basali per quel che riguarda il contenuto di metalli pesanti nelle specie considerate. Ciononostante, queste indagini sono a nostra conoscenza tra le poche che stimano i livelli di elementi al di fuori di stati di intossicazione, che per altro vengono valutati a livello ematico, e possono quindi essere considerate come un primo passo verso al definizione di valori di riferimento per gli elementi traccia e i metalli tossici; sono inoltre di notevole importanza per la valutazione dello stato di benessere degli animali.

Present work reports about the quantification of contaminants and thyroid hormones in blood of sea turtles from Southern Mediterranean Sea.

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We investigated the effect of five different diets and three volumes on the culture of Paracentrotus lividus larve (Echinodermata:Echinoidea), in controlled conditions. Larvae survivorship was proved to depend on the culture parameters applied. 1600 litre volumes and Nannochloropsis salina - Chaetoceros calcitrans diet produced more morphologically typical larvae and gave better results in terms of survivorship at metamophosis.

Aim of the workshop is to focus on the potential effects of pollutants on the health of marine megavertebrates. The workshop will focus on different aspects of natural and anthropogenic pollutants and on suspected or proved connections with specific/some pathologies. This workshop will be a valid chance to put together different competences and to allow an exchange on knowledge and opinions; it will as well present various ecopathological aspects in species which have a great value, also under the sociological point of view.

Heavy metals concentration was evaluated in tissues collected from stranded sunfish Mola mola from Northern Adriatic Sea. Obtained data show characteristic distribution of metals among tissues, which differs from that of other fish species. Present data are to our knowledge the first one concerning pollutants presence in this species.

Cervids have been used since a long time as biomonitoring tools, because of their ecological characteristics. An interesting matrix to bone for long term analysis of contaminants could be teeth, which represent an indication of life span accumulation of metals. Present work reports about heavy metals concentration in teeth of two species of cervids, roe deer (Capreolus capreolus) and red deer (Cervus elaphus) from two hunting areas of North-East Italy. Samples were collected from mandibles obtained from hunters and were analysed for heavy metals content by usin ICP-OES technique. Obtained data, reported as g/g ± s.e. dry weight, were analysed in order to identify any difference in heavy metals concentration between species and between sampling areas. Highest concentration were observed for Cr, followed by As and Cd. All other metals were close or below the limit of detection. No differences in heavy metals concentrations between sampling areas or species was observed. A similar deposition process can be considered for the two species, as well as a similar chronic exposure levels.

We report herein the results of post-mortem, parasitological, microbiological, histopathological, immunohistochemical (IHC), indirect immunofluorescence (IIF), biomolecular (RT-PCR) and serological investigations carried out on 8 (1 male calf, 1 subadult female, 5 adult females, 1 adult male) striped dolphins (S. coeruleoalba) found stranded from August to November 2007 on the Ligurian Sea coast of Italy. A (broncho-interstitial) pneumonia occurred in 6 animals, with numerous parasitic bodies in both pulmonary and extra-pulmonary locations. Histologically, a multifocal, subacute to chronic, nonpurulent meningo-encephalitis occurred in 1 adult male and 3 adult females, with more or less prominent perivascular cuffs of inflammatory mononuclear cells and peri-paravascular macrophage accumulations. A mild choroid plexus lymphocytic-plasmacytic infiltration was also present, along with moderate neuronal damage, although no viral inclusions were seen in any brain cell (nor in any other cell type). The blood sera from 4 dolphins (1 calf, 1 subadult, 2 adults) had anti-Morbillivirus neutralizing antibodies, with positive titres ranging from 1:10 to 1:40. Nevertheless, IHC, IIF and RT-PCR investigations gave negative results in all animals. On the basis of the above findings, we conclude that one or more factors other than Morbillivirus likely were the primary cause of these dolphin mortality episodes.

Presentation of GVP

Quinolones are widely used due to their high efficacy against Gram- positive and anaerobial bacteria. Anyway, among adverse effects of these drugs chondrotoxicity has been observed and recorded, thus leading to a reduced use in pediatric age. These drugs are also commonly used for the treatment of infectious diseases in chondroichtes, but little or no information is at present available concerning potential adverse effect of these active principles on the cartilage of selaceans. Present work reports about the evaluation of chondrotoxicity to Scyliorhinus canicola following intramuscular treatment with a quinolone, enrofloxacin. Three experimental groups were prepared: a control, a low dose and a high dose group, which were treated with saline, 5 mg/kg and 10 mg/kg enrofloxacin respectively, daily for a 14 days period, following normal therapeutic protocols (Smith et al., 2004). Animals were fed daily ad libitum and weight was regularly recorded every 3 days in order to detect for any toxicity. Blood samples were taken at day 0 and 14 of treatment and at day 14 post-treatment. Half of the animals were suppressed at the end of the 14 days treatment period, while all other animals were killed at the end of the 14 days post-treatment period. Ultrastuctural or densitometric alteration in cartilage structure was detected, as well as any changing in hematological parameters. Preliminary results are discussed in present work as well as the possibility to use small sharks as lab animals for this protocols.

The impact of morbillivirus on cetacean is well known since a long time. Indeed, since 1987, at least eight morbillivirus infection (MI) epidemics have caused mass mortality of several free-living pinniped and cetacean populations around the world. At post-mortem examination, a commonly occurring macroscopic lesion is represented by more or less severe bilateral pneumonia, with consolidation, congestion and oedema of both lungs, which fail to collapse. Among the still debated, or even controversial issues regarding MI in sea mammals, the one related to the origin of their causative agents is of particular concern. It has recently also debated if some episodes ascribed to MI were real morbillivirus infections, as serological or immunohistochemical techniques failed to identify the virus. Another intriguing issue regards the synergistic effects, if any, associated with chronic exposure to a number of environmental pollutants, such as organochlorines and heavy metals. In fact, it is also unknown whether and how these chemicals contribute towards modulating the pathogenic and pathogenetic activity primarily displayed by sea mammal morbilliviruses. Among others, heavy metals have proved to act as immunomodulators in many mammals species, mainly acting as immunosuprressor. In present study we report about heavy metals (Cd, Pb and Hg) concentration in liver and muscle of stranded dead stripped dolphin (n= 9) from Tirrenian Sea collected during a suspected MI episode. Sampled cohort included a 5 days old calf, two sub-adult and 6 adult subjects. Contemporarily, autoptic lesions were recorded and a serological survey was conducted on brain samples of 8 out of 9 animals in order to verify if the causative agent of death was morbillivirus. Any possible correlation with immunosuppressive action of metals was also tested, even if low number of animals did not allow any statistical analysis. Obtained results showed mean metals concentration reported in table 1. Lowest levels of all metals were observed in the calf, but it should be noted that 10 out of 18 samples were below the limit of detection (LOD) for lead. These levels were always below the immunotoxic thresholds for cadmium and lead, while were always well below the threshold for mercury. Thus a certain degree of immunosuppression, including increase of apoptosis, reduction in lymphocyte proliferation and fagocytic activity, could be considered for all animals. Serological survey revealed that only 4 out of 8 animals analysed were positive to morbillivirus. Interestingly, positive animals presented no typical morbillivirus brain lesion, which were present in negative subjects. It should be noted the presence among sero-positive animals of the calf. In this case, a mother-calf antibody transfer with colostrum should be considered. Giovanni, qui ti sarei molto grata se potessi scrivere tu qualcosa sulla sierologia, tipo la questione del passaggio col colostro al cucciolo, la possibilità di un'immunità acquisita a seguito di esposizione "cronica" al virus, i dubbi sull'effettiva esposizione a morbillivirus o ad altri patogeni. Interestingly, when metal data were evaluated on the basis of serological results, higher levels of metal were found in sero-negative animals, as evident in hepatic lead and mercury Obtained data seems to suggest the occurrence of an infection different from morbillivirus, which induce cerebral lesions similar to MI and whose morbidity is increased by the action of mercury and probably of other contaminants known to act as immunosuppressive agents. Further and deeper researches are considered necessary in order to better define which is the role of pollutants in the severity of epidemics and, more important, which could be the real causative agent of the die-off of 2007 in Italy.

Estimation of age in fish is fundamental to know the growth rate and other biological parameters. Here is presented a first effort to use eye lens weight and diameter in order to determine age in Silurus glanis. Eyes were obtained from a sample of Silurus collected in Campotto lakes and dried lenses were then measured in weight and diameter with a precision balance and digital calliper. Correlation with an estimation of age based on the vertebral body was calculated. The best results came from the weight as the diameter produced a less informative curve. No significant differences between genders were detected.