A modified culture medium for improved isolation of marine vibrios

Marine Vibrio members are of great interest for both ecological and biotechnological research, which often relies on their isolation. Whereas many efforts have been made for the detection of food-borne pathogenic species, much less is known about the performances of standard culture media toward environmental vibrios. We show that the isolation/enumeration of marine vibrios using thiosulfate-citrate-bile salts-sucrose agar (TCBS) as selective medium may be hampered by the variable adaptability of different taxa to the medium, which may result even in isolation failure and/or in substantial total count underestimation. We propose a modified TCBS as isolation medium, adjusted for marine vibrios requirements, which greatly improved their recovery in dilution plate counts, compared with the standard medium. The modified medium offers substantial advantages over TCBS, providing more accurate and likely estimations of the actual presence of vibrios. Modified TCBS allowed the recovery of otherwise undetected vibrios, some of which producing biotechnologically valuable enzymes, thus expanding the isolation power toward potentially new enzyme-producers Vibrio taxa. Moreover, we report a newly designed Vibrio-specific PCR primers pair, targeting a unique rpoD sequence, useful for rapid confirmation of isolates as Vibrio members and subsequent genetic analyses

Proportional Venn diagram and determinants of allergic respiratory diseases in Italian adolescents

Large variations in prevalence of atopy and allergic diseases are reported worldwide in children, but in epidemiological studies the use of skin prick tests (SPT) and spirometry along with questionnaires is not common in the Mediterranean Area. The present work was aimed at evaluating the prevalence of current asthma (CA), rhinoconjunctivitis (RC), and eczema (E), with atopy and respiratory function, and the role of risk factors for allergic respiratory diseases. A total of 2150 Italian schoolchildren were cross-sectionally investigated through respiratory questionnaire, SPT, and spirometry. A proportional Venn diagram quantified the distribution of CA, RC, and E, stratifying for allergic sensitization to show differences in prevalence of allergic diseases among subjects with and without positive SPT. CA prevalence was 4.2%, RC 17.9%, and E 5.3%. CA and RC increased, while E decreased, with respect to previous local studies. Allergic sensitization prevalence (evaluated as positive response to at least one SPT) was 39.2%. A double Venn diagram identified 15 categories. Atopic CA was threefold more frequent than non-atopic CA. Atopic vs non-atopic RC and E were 9.6% vs 10.3% and 2.0% vs 3.3%, respectively. Atopic vs non-atopic RC associated with CA were 1.6% vs 0.5%; the same figures for RC associated with E were 0.8% vs 1.3%. Asymptomatic atopic subjects were 27.0%. Atopy, RC, parental asthma, and environmental risk factors were associated with CA. Atopy and environmental factors were risk factors also for RC. Asthma and traffic exposure were linked to reduced lung function. Respiratory allergic diseases are still increasing and largely concomitant in Italian adolescents. Atopy is more important for CA than RC. Avoiding exposures to measured environmental risk factors would prevent 41% of current asthma and 34% of rhinoconjunctivitis

Characterization of translationally controlled tumour protein from the sea anemone Anemonia viridis and transcriptome wide identification of cnidarian homologues

Gene family encoding translationally controlled tumour protein (TCTP) is defined as highly conserved among organisms; however, there is limited knowledge of non-bilateria. In this study, the first TCTP homologue from anthozoan was characterised in the Mediterranean Sea anemone, Anemonia viridis. The release of the genome sequence of Acropora digitifera, Exaiptasia pallida, Nematostella vectensis and Hydra vulgaris enabled a comprehensive study of the molecular evolution of TCTP family among cnidarians. A comparison among TCTP members from Cnidaria and Bilateria showed conserved intron exon organization, evolutionary conserved TCTP signatures and 3D protein structure. The pattern of mRNA expression profile was also defined in A. viridis. These analyses revealed a constitutive mRNA expression especially in tissues with active proliferation. Additionally, the transcriptional profile of A. viridis TCTP (AvTCTP) after challenges with different abiotic/biotic stresses showed induction by extreme temperatures, heavy metals exposure and immune stimulation. These results suggest the involvement of AvTCTP in the sea anemone defensome taking part in environmental stress and immune responses

Esperimento pilota mirato all’analisi dell’effetto della stimolazione immunitaria su A. viridis, finalizzato alla produzione di composti biologicamente attivi

L’esperimento mira a valutare l’eventuale attività antibatterica di peptidi prodotti da individui di Anemonia viridis in seguito a stimolazione immunitaria e quindi alla realizzazione di un protocollo riproducibile e ottimizzato per la produzione di tali composti

Applicazione di un protocollo di immuno-stimolazione su individui di Pracambarus clarkii mirato alla produzione di peptidi con attività biologica

Uno dei principali problemi inerenti l’identificazione e l’analisi di nuovi peptidi naturali di origine marina è la disponibilità in termini quantitativi del materiale bioattivo (Cragg et al., 1997). Infatti, le concentrazioni di tali peptidi negli invertebrati marini sono spesso meno del 10-6 % del peso umido, ed il loro rendimento a seguito delle procedure di estrazione tradizionali risulta ancora più basso. Tuttavia, sono molteplici i fattori che possono influenzare l'esito della identificazione degli Anti Microbial Peptides (AMP), come le variazioni geografiche e stagionali (sito di campionamento), le diverse fasi della vita (specie pelagiche o bentoniche), l'età, il sesso e lo stato fisiologico (malattia, allevamento, muta). A causa della difficoltà nell’ottenere quantità sufficienti di composti bioattivi, potrebbe essere più produttivo dissezionare in diversi tessuti/organi gli invertebrati selezionati per poi estrarre il materiale da saggiare (Haug et al., 2002a, 2002b, 2004) ciò al fine di separare i peptidi di interesse da migliaia di sostanze inattive. Dividere gli organismi in parti differenti selezionando ciascun tessuto potrebbe anche fornire l'indicazione se l'animale produce da sé il principio attivo o se esso proviene dalla dieta o ancora è associato a parassiti o microrganismi. La maggior parte degli AMP marini sono stati di fatto isolati dal compartimento sangue, sia dall’emolinfa/fluido celomatico (sangue intero) o dagli emociti/celomociti (cellule del sangue). Alcuni peptidi tuttavia sono stati scoperti e isolati da altri tessuti, ma essendo questi circondati (negli invertebrati marini) dagli emociti/celomociti, è possibile che provengano da questi ultimi. Inoltre tutte le variabili analitiche hanno una potenziale influenza sul risultato sia in termini quantitativi che qualitativi oltre che per la stabilità del campione anche per la riproducibilità del protocollo. Tra queste variabili ricadono le condizioni di conservazione del campione (tempo trascorso dall’estrazione, la temperatura prima dell’estrazione ed alla separazione, il congelamento e la liofilizzazione), la tipologia del campione ed il metodo di prelievo, la separazione delle cellule dal sangue (velocità di centrifugazione, durata e temperatura), l’uso del tampone (tipo, pH, forza ionica e temperatura), l'uso e il tipo di anticoagulanti e cocktail inibitori della proteasi. Inoltre, dati bibliografici riportano come la stimolazione immunitaria determini un aumento sia quantitativamente che qualitativamente della produzione di peptidi antimicrobici. Per tale motivo si è proceduto ad effettuare un esperimento pilota per valutare se in tali condizioni fosse superiore la quantità di piccoli peptidi sintetizzati rispetto ad individui non stimolati e se nei differenti tempi di stimolazione selezionati, emergesse una differenziazione a carico dei peptidi prodotti in relazione ad una possibile attività di natura antibatterica

Gene expression changes after parental exposure to metals in the sea urchin affect timing of genetic programme of embryo development

It is widely accepted that phenotypic traits can be modulated at the epigenetic level so that some conditions can affect the progeny of exposed individuals. To assess if the exposure of adult animals could result in effects on the offspring, the Mediterranean sea urchin and its wellcharacterized gene regulatory networks (GRNs) was chosen as a model. Adult animals were exposed to known concentrations of zinc and cadmium (both individually and in combination) for 10 days, and the resulting embryos were followed during the development. The oxidative stress occurring in parental gonads, embryo phenotypes and mortality, and the expression level of a set of selected genes, including members of the skeletogenic and endodermal GRNs, were evaluated. Increased oxidative stress at F0, high rates of developmental aberration with impaired gastrulation, in association to deregulation of genes involved in skeletogenesis (dri, hex, sm50, p16, p19, msp130), endodermal specification (foxa, hox11/13b, wnt8) and epigenetic regulation (kat2A, hdac1, ehmt2, phf8 and UBE2a) occurred either at 24 or 48 hpf. Results strongly indicate that exposure to environmental pollutants can affect not only directly challenged animals but also their progeny (at least F1), influencing optimal timing of genetic programme of embryo development, resulting in an overall impairment of developmental success

Obesity and iron deficiency anemia as risk factors for asymptomatic bacteriur

Background: Few studies examined the risk factors of asymptomatic bacteriuria, showing contradictory results. Our study aimed to examine the association between different clinical and laboratory parameters and asymptomatic bacteriuria in internal medicine patients. Materials and methods: 330 consecutive hospitalized subjects, asymptomatic for urinary tract infections (UTIs), underwent to microscopic examination of urine specimens. 100 subjects were positive for microscopic bacteriuria and were recruited into the study. At the quantitative urine culture 31 subjects of study population were positive while 69 subjects were negative for bacteriuria. Results: The analysis of clinical characteristics showed that the two groups of subjects (positive and negative urine culture for bacteriuria) were significant different (p b 0.05) about obesity (76.7% vs 42% respectively), metabolic syndrome (80.6% vs 44,9%), cholelithiasis (35.5% vs 13,2%) and iron deficiency anemia (80.6% vs 53,6%). The univariate analysis showed that only obesity, cholelithiasis and iron deficiency anemia were positively associated with positive urine culture for bacteriuria (Odds Ratios [OR] = 3.79, p = 0.0003; OR = 2,65, p =0.0091; OR = 2.63, p = 0.0097; respectively). However, the multivariate analysis by logistic regression showed that only obesity and iron deficiency anemia, independently associated with positive urine culture for bacteriuria (OR = 3.9695, p = 0.0075; OR = 3.1569, p = 0.03420 respectively). Conclusions: This study shows that obesity and iron deficiency anemia are independent risk factors for asymptomatic bacteriuria

Rapporto tecnico sulla valutazione della biomassa ittioplanctonica mediante l'utilizzo del Multi Plankton Sampler (MPS)

Il piano di campionamento della campagna oceanografica BANSIC'14, condotta a bordo della N/O "URANIA" dal 22 Luglio al 9 Agosto 2014, lungo transetti sotto costa e a largo delle coste meridionali della Sicilia, ha avuto l’obiettivo generale dello studio delle relazioni tra le strutture oceanografiche a mesoscala (vortici verticali ed orizzontali, upwelling, etc.) e le strutture spaziali dei fenomeni biologici relativi ai primi anelli della catena trofica (zooplancton, distribuzione e abbondanza di larve di piccoli pelagici e grandi pelagici) (vedi Rapporto finale BANSIC 2014) per la stima dell’abbondanza dello stock riproduttore. E' una campagna di ricerca nell'ambito del WP3 del progetto SSD-Pesca, finanziato dal MIUR su fondi MISE, a supporto della pesca italiana nelle Regioni Obiettivo 1 e del progetto RITMARE (SP2_WP4_AZ2_UO04). Il campionamento ittioplanctonico è inserito anche nel piano di lavoro del progetto regionale MIPAF-FAO “MedSudMed” (“Assessment and Monitoring of the Fishery Resources and the Ecosystems in the Straits of Sicily”). Il campionamento dell’ittioplancton, durante questa campagna, oltre ai metodi tradizionali quali le reti di tipo Bongo, ha visto l'utilizzo del Multi Plankton Sampler (MPS) MultiNet. I campionatori ittioplanctonici hanno lo scopo di prelevare porzioni di mesozooplancton da un massimo di 100 m fino alla superficie, in quanto le uova di pesci pelagici possiedono una galleggiabilità tale che nonostante le turbolenze superficiali dell’acqua, un campionamento entro i primi metri restituisce un dato affidabile della distribuzione anche se alcune uova possono trovarsi a maggiore profondità (Ahlstrom, 1959). Il campionatore MPS consente, a differenza di altri strumenti, di prelevare la frazione di zooplancton d'interesse con diverse modalità di campionamento: orizzontale, verticale e obliquo e, allo stesso tempo, permette di campionare a differenti quote di profondità. Le informazioni così ottenute sono state utilizzate per valutare la variazione della biomassa ittioplanctonica, l'abbondanza e la composizione delle specie lungo gli strati della colonna d'acqua anche in relazione alle componenti oceanografiche. Grazie all'utilizzo di questo strumento è possibile validare e verificare alcune informazioni e acquisirne delle nuove sull’ecologia delle specie larvali e sul mesozooplancton, sul modo in cui queste si distribuiscono lungo la colonna d'acqua e sulle interazioni intra ed interspecifiche legate anche a fattori oceanografici. Ciò consente di ottenere maggiori informazioni e contribuire al miglioramento della comprensione della biologia e dell'ecologia delle specie rinvenute e, nel contempo, approfondire e migliorare le conoscenze sugli stadi di sviluppo di uova di specie ittiche che allo stadio embrionale sono ancora poco conosciuti

Seascape connectivity of European anchovy in the Central Mediterranean Sea revealed by weighted Lagrangian backtracking and bio-energetic modelling

Ecological connectivity is one of the most important processes that shape marine populations and ecosystems, determining their distribution, persistence, and productivity. Here we use the synergy of Lagrangian back-trajectories, otolith-derived ages of larvae, and satellite-based chlorophyll-a to identify spawning areas of European anchovy from ichthyoplanktonic data, collected in the Strait of Sicily (Central Mediterranean Sea), i.e., the crucial channel in between the European and African continents. We obtain new evidence of ecosystem connectivity between North Africa and recruitment regions off the southern European coasts. We assess this result by using bio-energetic modeling, which predicts species-specific responses to environmental changes by producing quantitative information on functional traits. Our work gives support to a collaborative and harmonized use of Geographical Sub-Areas, currently identified by the General Fisheries Commission for the Mediterranean. It also confirms the need to incorporate climate and environmental variability effects into future marine resources management plans, strategies, and directives

Moving Toward a Strategy for Addressing Climate Displacement of Marine Resources: A Proof-of-Concept

Realistic predictions of climate change effects on natural resources are central to adaptation policies that try to reduce these impacts. However, most current forecasting approaches do not incorporate species-specific, process-based biological information, which limits their ability to inform actionable strategies. Mechanistic approaches, incorporating quantitative information on functional traits, can potentially predict species- and population-specific responses that result from the cumulative impacts of small-scale processes acting at the organismal level, and can be used to infer population-level dynamics and inform natural resources management. Here we present a proof-of-concept study using the European anchovy as a model species that shows how a trait-based, mechanistic species distribution model can be used to explore the vulnerability of marine species to environmental changes, producing quantitative outputs useful for informing fisheries management. We crossed scenarios of temperature and food to generate quantitative maps of selected mechanistic model outcomes (e.g., Maximum Length and Total Reproductive Output). These results highlight changing patterns of source and sink spawning areas as well as the incidence of reproductive failure. This study demonstrates that model predictions based on functional traits can reduce the degree of uncertainty when forecasting future trends of fish stocks. However, to be effective they must be based on high spatial- and temporal resolution environmental data. Such a sensitive and spatially explicit predictive approach may be used to inform more effective adaptive management strategies of resources in novel climatic conditions