IDENTIFICATION OF ALEXANDRIUM (DINOPHYCEAE) SPECIES USING PCR AND rDNA-TARGETED PROBES

A PCR (polymerase chain reaction)-based assay for the detection of Alexandrium species in cultured samples using rDNA-targeted probes was developed. The internal transcribed spacers 1 and 2 (ITS1 and ITS2) and the 5.8S ribosomal RNA gene (rDNA) from cultured isolates of A. tamarense (Lebour) Taylor, A. catenella (Whedon et Kofoid) Balech, A. fundyense Balech and A. lusitanicum Balech were amplified using PCR and sequenced. Sequence comparisons showed that the 5.8S and ITS1-ITS2 regions contain sequences specific for the Alexandrium genus, especially at the 3' end of the 5.8S coding region. PCR primers and a radioactive ³²P-labeled DNA probe were devised for this region. The cross-reactivity of the PCR primers and probe was tested against cultured isolates of Alexandrium and other dinoflagellates and diatoms. All the Alexandrium isolates screened reacted toward the genus-specific probe; in contrast, the other groups of microalgae (dinoflagellates and diatoms) did not react with the probe. Furthermore, the PCR amplification technique combined with the use of the rDNA-target probe allowed us to develop a method for the detection of Alexandrium cells in cultured samples. This PCR method might offer a new approach for the identification and enumeration of the HAB (harmful algal bloom) species present in natural phytoplankton populations.     

Cage culture effects on mullets (Mugilidae) liver histology and blood chemistry profile

A comparative study of blood chemistry and histology was conducted on two groups of mullets (Mugilidae) living under different conditions with different feed sources. The aquaculture influenced mullet group (AIM), was collected near fish farms and the control group of mullet (CM) was caught in the waters without any aquaculture activities. Histological and biochemical procedures were employed to study liver histomorphology, plasma aspartate and alanine aminotransferase (AST, ALT), triglyceride (TRIG), cholesterol (CHOL), glucose (GLU) and total protein (TP) of both AIM and CM. Moderate histological changes (lipid infiltration) were observed in the liver of AIM. Significant changes in plasma variables were observed in AIM. Blood chemistry variables measured proved to be good indicators of artificial feed effects. Classical statistical approaches were applied to the blood chemistry and histopathology data. For the first time machine learning techniques were used to generate comprehensible classification models and to explore blood chemistry variable importance, strength, their mutual interactions or dependencies, and to investigate reliability of particular variables within the groups.     

FUnaria mediterranea monstruositäten aus italien

Abstract

The aim of the present study is to demonstrate the value of some nitrophilous plants as bioindicators related to global warming. As a case study, the space–time dynamics of the populations of some nitrophilous and ruderal species have been investigated along the southern-central Adriatic side of the Italian Peninsula. These have been examined according to the range of their distribution areas, to their ecological needs, and to the availability of past data, which have then been related to the data on global warming across the same territories. The choice for this investigation was for nitrophilous species with a Mediterranean distribution – of the Stenomediterranean type – and for some recent entries into the local flora as alien species. The spread of these species occurs in areas with intense human activity, where they have exploited the conditions of the warmer niche, such that their presence and their spatial spread observed over time are clearly linked to global warming. For all of the species in question, rapid increases in the population numbers have been observed, along with a northward shift of their distribution areas. These changes correspond to the increase in average annual temperature as revealed by the thermometric measurements.     

The benthic algal flora on rocky substrata of the Tremiti Islands (Adriatic Sea)

ABSTRACT

The results of a floristic study of benthic marine macroalgae on rocky substrata from the Tremiti Islands (Adriatic Sea) are presented. The list of taxa at specific and infraspecific level consists of 226 Rhodophyceae, 59 Fucophyceae and 36 Chlorophyceae. Of these 321 taxa, 147 are newly reported from the Tremiti Islands. Data on the reproductive phenology of each species, and comments on some species of particular interest are also given.     

Cartography of littoral rocky-shore communities (CARLIT) as a tool for ecological quality assessment of coastal waters in the Eastern Adriatic Sea

Littoral cartography is a valuable tool which uses shallow water macroalgal communities for monitoring ecosystem health and water quality. Cartography of littoral rocky-shore communities (CARLIT) is a European Union Water Framework Directive-compliant monitoring method widely used in the Western Mediterranean Sea. This non-destructive method is based on a visual observation of the type and length of coast occupied by rocky-shore communities in the upper-sublittoral zone. Here we present the first wide-scale application of CARLIT in the Adriatic Sea. The reference values calculated specifically for the Croatian coast (Eastern Adriatic Sea) ensure accurate calculation of the ecological status. The measured ecological quality ratio values (EQR) show a good linear relation with the Land Uses Simplified Index (LUSI) and appropriately reflect an increase in nutrient concentrations. The results suggest that this method can be used as a reliable biomonitoring tool in the Eastern Adriatic Sea. This study highlighted that Cystoseira forests thrive in 39% of the surveyed coastline and 23% of the coastline seems to be subjected to overgrazing by sea urchins. Our results provide a spatially accurate information on the abundance of different upper-sublittoral communities and the associated ecological status, which can be used in future management plans for improving water quality.     

The Late Glacial–Holocene record from Central Adriatic Sea: Paleoenvironmental reconstruction based on benthic foraminiferal assemblages

New micropaleontological data coming from three cores collected on the meso-Adriatic continental shelf (Vasto area) are studied. Comparisons to foraminiferal assemblages and radiocarbon dates previously collected from cores in the San Benedetto del Tronto and Tremiti areas allow the correlation of patterns observed in shallow water areas with those in deeper parts of the basin. We focused on the response of benthic foraminifera during the Holocene high-stand, corresponding to the installation of the recent sedimentary and trophic system. An influence of anthropogenic impacts cannot be ruled out; its effects consist of a depletion of oxygen level with a consequent modification of the structure of benthic foraminiferal assemblages. During the glacial/post glacial cycle, three phases, characterized by a total of six foraminiferal biofacies including different species assemblages were recognized. The first phase, from 14 kyr BP to 11 kyr BP, corresponds to the Bölling/Alleröd and Younger Dryas cold event, before the Holocene sea-level rise. During this phase, the continental shelf was characterized by an infralittoral environment with productive waters owing to the proximity of the Po river delta at the edge of the Mid-Adriatic Deep. The second phase, from 11 kyr BP to 4 kyr BP, represents the Holocene sea-level rise and is characterized by a condensed sedimentation spread over the entire basin. The third phase corresponds to the Holocene high-stand, during which time the modern current system became established. During this phase, the eastern portion of the shelf underwent to the central part of the mud-belt, corresponding to the sub-recent conditions. Recent eutrophication resulting from human activities over the last few centuries is evidenced by frequency fluctuations of typically opportunistic taxa such as Nonionella turgida and Epistominella vitrea.     

The evolution and phytoplankton composition of mucilaginous aggregates in the northern Adriatic Sea

Recurrent occurrences of visible mucilage “clouds” that cover areas up to several hundred kilometres with vertical dimensions of 20–30 m have been recorded in the stratified water column in the northern Adriatic. In the past this was described as “mare sporco” phenomenon. Past studies of the phenomenon indicated that phytoplankton is an important component of mucilage. Our research revealed the composition of phytoplankton assemblages in different types of mucilaginous aggregates collected during the summers of 1997 and 2000 using pigment biomarkers (HPLC). Phytoplankton biomass in the mucilage samples was very high, ranging from 7.9 μg g⁻¹ to 390.8 μg g⁻¹ of chlorophyll a per unit of dry mass of mucilage. The phytoplankton community in the early, loose stage of mucilaginous aggregates was heterogeneous, as indicated by the diversity of detected pigments. The number of phytoplankton groups decreased as the aggregates aged and diatoms increased in relative biomass (up to 92.7%). Phytoplankton biomass in seawater was similar in years with and without mucilage; however, significantly higher contributions to the total biomass of 19′-hexanoyloxyfucoxanthin-containing phytoplankton (prymnesiophytes) were found in the upper 10 m in spring of the “mucilaginous years” (1997 and 2000) followed by prevalence of diatoms in summer. The F_p pigment index used to assess seawater trophic conditions reached lower values in April–May in mucilaginous years (1997 and 2000) compared to non-mucilaginous years (1998 and 1999). We conclude that the role of prymnesiophytes and other small flagellates is crucial for the initial phases of mucilage appearance. Aggregates represent a favourable environment for the secondary development of opportunistic diatoms that foster mucilage formation.     

Seasonal and vertical distribution of the ciliated protozoa and micrometazoa in Kaštela Bay (central Adriatic)

Seasonal and vertical distribution of tintinnids, non-loricate ciliates and micrometazoa were studied in Kaštela Bay (central Adriatic Sea) throughout 1995. The species composition of tintinnids and copepods were studied as well. This is the first estimation of non-loricate ciliate biomass in the coastal area of the central Adriatic. Non-loricate ciliates were quantitatively the best represented ciliated protozoa, whereas nauplii were the most numerous micrometazoan organisms. Temperature affected the distribution of most micrometazoan components of the zooplankton and that of non-loricate ciliates. The temperature-dependent presence of individual size categories of non-loricate ciliates was also established. Apart from the interaction between microzooplankton groups, the influence of biotic factors, such as phytoplankton, bacteria, non-pigmented nanoflagellates (NNF) and mesozooplankton, was also discussed. The abundance of ciliates was controlled by both food supply (phytoplankton and NNF) and micrometazoan grazing. The results point to very complex trophic relationships within the planktonic community, suggesting that microzooplankton could be an important link between the microbial food web and higher trophic levels. Received in revised form: 8 November 2000 Electronic Publication     

Correlation between the presence of Gonyaulax fragilis (Dinophyceae) and the mucilage phenomena of the Emilia-Romagna coast (northern Adriatic Sea)

The formation of massive amounts of suspended mucilaginous organic matter which periodically affects the Adriatic Sea, has been regarded as a complex physico-chemical phenomenon resulting from the production of extracellular material by phytoplankton. Although the exact cause has remained obscure, the mechanism of its formation has usually been considered to be a long-term process, starting after the late winter-early spring blooms, and involving the participation of various algal species, mainly within the diatom group. In this paper we report on the results of a phytoplankton monitoring programme in northern Adriatic seawaters off the Emilia-Romagna coast of Italy which revealed the constant concomitant presence of the dinoflagellate Gonyaulax fragilis (Schütt) Kofoid and mucilaginous formations. In the early stages of the phenomenon the dinoflagellate was clearly observable by microscopic examination in the mucilage, but as the mucilage aged this alga almost completely decomposed and diatom cells increased in number and became predominant. Although characterized by a slow growth rate in culture, in natural seawater G. fragilis was observed to reach cell densities of up to 7.0×10⁶ cells l⁻¹. The results of this study lead us to propose the hypothesis that the appearance of mucilage in the water column of the Adriatic Sea is the consequence of a seasonal growth of this dinoflagellate favoured by specific environmental circumstances.