Levels of PSP toxins in bivalves exposed to natural blooms of Alexandrium minutum in Catalan harbours

The development, validation, comparison and evaluation of analytical methods for marine toxins rely on the availability of toxic material. Within the project JACUMAR PSP, our interest is mainly focused on autochthonous bivalve species with the toxic profile of Alexandrium minutum, since this is the principal species involved regionally in PSP outbreaks. Mussels and oysters were exposed during few days in the harbor of Vilanova i la Geltrú, to blooms reaching a maximum A. minutum concentration of 200,000 cells L-1 in 2008, and 40,000 and 800,000 cells L-1, in 2009. Mussels, oysters and clams were exposed to one bloom of 22,000 cells L-1 in the harbor of Cambrils in 2009. In all situations higher toxic levels analyzed by HPLC-FD with postcolumn oxidation were observed in mussels (i.e. 1,200-2,500 μg eq. STX kg-1) than in oysters (i.e. 60-800 μg eq. STX kg-1) exposed to the same bloom. Blooms with higher concentrations of A. minutum did not correspond to higher levels of PSP toxins in bivalves. These differences may be explained by differences in A. minutum population dynamics, toxin production or in the physiological state or behaviour of shellfish. These results confirm that mussels concentrate more PSP toxins from A. minutum than oysters and clams.

A new local variety of Crepidomanes minutum (Hymenophyllaceae) in the Mascarene Archipelago (Indian Ocean) and a new record for Mauritius

Crepidomanes minutum (Hymenophyllaceae) is here identified and recorded from Mauritius for the first time. The Mauritian specimens, in addition to those of La Reunion observed at low to middle elevations, are easily distinguished from populations observed outside the Mascarene Archipelago by their dwarfed size and rarity of the stipe proliferation that usually characterizes this species. We thus describe a new variety in this species for the Mascarene Islands.

Exposure to toxic Alexandrium minutum activates the detoxifying and antioxidant systems in gills of the oyster Crassostrea gigas

Harmful algal blooms of Alexandrium spp. dinoflagellates regularly occur in French coastal waters contaminating shellfish. Studies have demonstrated that toxic Alexandrium spp. disrupt behavioural and physiological processes in marine filter-feeders, but molecular modifications triggered by phycotoxins are less well understood. This study analyzed the mRNA levels of 7 genes encoding antioxidant/detoxifying enzymes in gills of Pacific oysters (Crassostrea gigas) exposed to a cultured, toxic strain of A. minutum, a producer of paralytic shellfish toxins (PST) or fed Tisochrysis lutea (T. lutea, formerly Isochrysis sp., clone Tahitian (T. iso)), a non-toxic control diet, in four repeated experiments. Transcript levels of sigma-class glutathione S-transferase (GST), glutathione reductase (GR) and ferritin (Fer) were significantly higher in oysters exposed to A. minutum compared to oysters fed T. lutea. The detoxification pathway based upon glutathione (GSH)-conjugation of toxic compounds (phase II) is likely activated, and catalyzed by GST. This system appeared to be activated in gills probably for the detoxification of PST and/or extra-cellular compounds, produced by A. minutum. GST, GR and Fer can also contribute to antioxidant functions to prevent cellular damage from increased reactive oxygen species (ROS) originating either from A. minutum cells directly, from oyster hemocytes during immune response, or from other gill cells as by-products of detoxification.

Rapid detection and quantification of the marine toxic algae, Alexandrium minutum, using a super-paramagnetic immunochromatographic strip test

The dinoflagellates of Alexandrium genus are known to be producers of paralytic shellfish toxins that regularly impact the shellfish aquaculture industry and fisheries. Accurate detection of Alexandrium including A. minutum is crucial for environmental monitoring and sanitary issues. In this study, we firstly developed a quantitative lateral flow immunoassay (LFIA) using super-paramagnetic nanobeads for A. minutum whole cells. This dipstick assay relies on two distinct monoclonal antibodies used in a sandwich format and directed against surface antigens of this organism. No sample preparation is required. Either frozen or live cells can be detected and quantified. The specificity and sensitivity are assessed by using phytoplankton culture and field samples spiked with a known amount of cultured A. minutum cells. This LFIA is shown to be highly specific for A. minutum and able to detect reproducibly 105 cells/L within 30 min. The test is applied to environmental samples already characterized by light microscopy counting. No significant difference is observed between the cell densities obtained by these two methods. This handy super-paramagnetic lateral flow immnunoassay biosensor can greatly assist water quality monitoring programs as well as ecological research.

Changes in the annual harmful algal blooms of Alexandrium minutum : effects of environmental conditions and drainage basin inputs in the Rance estuary (Brittany, France)

Time series of physico-chemical data and concentrations (cell L-1) of the toxic dinoflagellate Alexandrium minutum collected in the Rance macrotidal estuary (Brittany, France) were analyzed to understand the physico-chemical processes of the estuary and their relation to changes in bloom development from 1996 to 2009. The construction of the tidal power plant in the north and the presence of a lock in the south have greatly altered hydrodynamics, blocking the zone of maximum turbidity upstream, in the narrowest part of the estuary. Alexandrium minutum occurs in the middle part of the estuary. Most physical and chemical parameters of the Rance estuary are similar to those observed elsewhere in Brittany with water temperatures between 15–18 °C, slightly lowered salinities (31.8–33.1 PSU), low river flow rates upstream and significant solar radiation (8 h day-1). A notable exception is phosphate input from the drainage basin which seems to limit bloom development: in recent years, bloom decline can be significantly correlated with the decrease in phosphate input. On the other hand, the chemical processes occurring in the freshwater-saltwater interface do not seem to have an influence on these occurrences. The other hypotheses for bloom declines are discussed, including the prevalence of parasitism, but remain to be verified in further studies.

Development of Trichogramma australicum Girault (Hymenoptera : Trichogrammatidae) in Helicoverpa (Lepidoptera : Noctuidae) host eggs

Morphological studies of development of the egg parasitoid Trichogramma australicum Girault in the cotton bollworm, Helicoverpa armigera (Hubner), were conducted to provide benchmarks for assessing developmental rates in both natural hosts and artificial diets. Observations of living embryos and histological sections show that embryos proceed rapidly through cleavage and blastoderm formation and show a characteristic pinching or rotation 8 h after deposition. Eggs progressively increase in volume, primarily by increasing in diameter at the widest point. At 29 rectangle 1 C the duration of the egg stage is 22-24 h, the larval stage 27 h, the prepupal stage 50-52 h, and pupa 85 h. Larvae undergo dramatic shape changes as they ingest food but do not show signs of larval moults, reinforcing observations that there is only one larval instar. Criteria for staging the embryonic and postembryonic development in natural hosts will be used for future studies aimed at developing and refining artificial diets for Trichogramma.

Les Périssodactyles (Mammalia) du Miocène de Lisbonne

New data led us to revise Miocene Perissodactyla from Lisbon. Equids (Anchitherium) have been decribed by Alberdi & González (1999); we fully agree with their conclusions. On the other hand, the only Chalicotheriid cannot be clearly reported to any genus. It therefore will be left in open nomenclature Schizorheriinae ind.). The Rhinocerotids have been described in detail (1983). We could recognize now that: Protaceratherium sagicum is a synonym of P. minutum; Plesiaceratherium platyodon and Plesiaceratherium lumiarense should be ascribed to the genus Plesiaceratherium and not to Aceratherium; there are no reasons for name changes as far as the forms previously referred as Diaceratherium aurelianensis, Prosantorhinus germanicus and Hispanotherium matritensis are concerned; as the genera Dicerorhinus and Lartetotherium are distinct, Dicerorhinus (Lartetotherium) sansaniensis has to be named Lartetotherium sansaniensis; as Gaindatherium (Ïberotherium) rexmanueli cannot be reported to Gaindatherium, we therefore upgrade the subgenus Iberotherium to the genus'rank - hence the names of the concerned taxa become Iberotherium rexmanueli rexmanueli and I. rexmanueli zbyszewski; Chilotherium ibericus is but a dental variation of I. rexmanueli zbyszewski. The stratigraphic distribution and age of the Miocene Perissodactyla so far known in the Lower Tagus basin / Lisbon region are presented.

Contribution à l'étude des infusoires parasites du coecum de l'hydrocheire (Hydrocheirus capybara L.): I. La famille des cycloposthiidae

Os autores referem inicialmente que os ciliados da capivara (Hydrochoerus capibara L.) são conhecidos pelos estudos de MARQUES DA CUNHA & MUNIZ bem como de FLÁVIO DA FONSECA. Fazem o estudo em material recentemente colhido em viagem ao Brasil e agradecem às pessoas que facilitaram êstes estudos. Dizem das técnicas usadas na coleta e estudo do material. Trabalhando o material colecionado em uma capivara sacrificada em Ribeirão Prêto estudam os representantes da família Cycloposthidae. Dividem as espécies do gênero Cycloposthium em três secções: a primeira com C. hydrochoeri Cunha, 1915; a segunda com C. minutum Cunha & Muniz, 1927; C. lenticularis sp. n. e C. elongatum sp. n.; a terceira com C. caudatum Cunha & Muniz, 1927 e C. compressum Cunha 1915 incluidas em novo subgênero: Diplolophus. Estudam pormenorisadamente C. hydrochoeri fazendo numerosas observa��ões da morfologia em exemplares de prepara��ão total, bem como em cortes. Na segunda secção dão a diagnose das 3 espécies nela incluídas, das quais duas novas para a ciência: C. lenticularis, com corpo quase tão largo como longo, 40 a 60 μ de comprimento por 40 a 45 μ de largura e processos caudais muito curtos ora acuminados ora arredondados. C. elongatum, de tamanho relativamente grande, medindo de comprimetno 160 a 200 μ por 60 a 90 μ de largura. Para a terceira secção estabelecem um novo subgênero - Diplolophus - com D.(D.) compressum e D.(D.) caudatum. O subgênero é caracterisado, principalmente, por serem os cirros adorais distribuídos em duas espirais: uma interna e outra mais externa. Finalisando o trabalho fazem comentários sôbre os componentes da família Cycloposthidae.

A new species of Limnoderetrema (Trematoda, Digenea) from the freshwater Atherinid fish Basilichthys australis Eigenmann, 1927 from the south of Chile

Limnoderetrema tolosai sp. n. (Trematoda, Digenea) within an autochthonous freshwater fish, the silverside Basilichthys autralis Eigenmann, 1927, from Lakes Riñihue and Ranco in Southern Chile is described. The species is distinguished from Limnoderetrema minutum (Manter, 1954) by the presence of one spine in the cirrus and cecal bifurcation nearer to ventral sucker than to pharinx. It is proposed Limnoderetrema macrophallus (Szidat & Nani, 1951) n. comb. (originally Steganoderma). Limnoderetrema tolosai differs from L. macrophallus since it cirrus has a distal spine and by its vitelline follicles distribution. It seems that Limnoderetrema spp. of South America are highly specific unlike L. minutum of New Zealand.