An Ins(1,4,5)P3 receptor in Paramecium is associated with the osmoregulatory system

In the ciliate Paramecium, a variety of well characterized processes are regulated by Ca2+, e.g. exocytosis, endocytosis and ciliary beat. Therefore, among protozoa, Paramecium is considered a model organism for Ca2+ signaling, although the molecular identity of the channels responsible for the Ca2+ signals remains largely unknown. We have cloned - for the first time in a protozoan - the full sequence of the gene encoding a putative inositol (1,4,5)-trisphosphate (Ins(1,4,5)P3) receptor from Paramecium tetraurelia cells showing molecular characteristics of higher eukaryotic cells. The homologously expressed Ins(1,4,5)P3-binding domain binds [3H]Ins(1,4,5)P3, whereas antibodies unexpectedly localize this protein to the osmoregulatory system. The level of Ins(1,4,5)P3-receptor expression was reduced, as shown on a transcriptional level and by immuno-staining, by decreasing the concentration of extracellular Ca2+ (Paramecium cells rapidly adjust their Ca2+ level to that in the outside medium). Fluorochromes reveal spontaneous fluctuations in cytosolic Ca2+ levels along the osmoregulatory system and these signals change upon activation of caged Ins(1,4,5)P3. Considering the ongoing expulsion of substantial amounts of Ca2+ by the osmoregulatory system, we propose here that Ins(1,4,5)P3 receptors serve a new function, i.e. a latent, graded reflux of Ca2+ to fine-tune [Ca2+] homeostasis.

Characterization of Acanthamoeba-microsphere association by multiparameter flow cytometry and confocal microscopy

Background: Acanthamoebae, in common with other protozoa, readily endocytose particulate material, which in turn may lead to the spread of infectious disease. Methods: Evaluation and quantification of plain and carboxylate FITC-microsphere association with acanthamoebal trophzoites was undertaken using a combination of flow cytometry and confocal microscopy. Trophozoites from strains and species of Acanthamoeba were exposed to plain and carboxylate FITC-microspheres. Microsphere size and aspects such as trophozoite starvation, maturity, and exposure to metabolic inhibitors were assessed. Results: All species and strains of Acanthamoeba readily endocytosed plain and carboxylate microspheres. Starving trophozoites significantly increased binding and potential ingestion of microspheres, whereas trophozoites of increasing maturity lost such abilities. Trophozoites showed a significant preference for 2.0- and 3.0-μm-diameter microspheres when compared with other sizes, which in turn could occupy much of the cytoplasm. The physiological inhibitors sodium azide, 2,4-clinitrophenol, and cytochalasin B reduced microsphere association with trophozoites; however, some microspheres still bound and associated with trophozoites after inhibitor exposure, a manifestation of both active and inactive agent involvement in microsphere endocytosis. Conclusions: Even though the origins of microsphere binding by acanthamoebal trophozoite remains shrouded, the combination of flow cytometry and confocal microscopy supported synergistic quantification and qualification of trophozoite-microsphere endocytosis. © 2006 International Society for Analytical Cytology.

Glycogen Synthase Kinase 3 influences cell motility and chemotaxis by regulating phosphatidylinositol 3 kinase localization in Dictyostelium discoideum

Glycogen Synthase Kinase 3 (GSK3), a serine/threonine kinase initially characterized in the context of glycogen metabolism, has been repeatedly realized as a multitasking protein that can regulate numerous cellular events in both metazoa and protozoa. I recently found GSK3 plays a role in regulating chemotaxis, a guided cell movement in response to an external chemical gradient, in one of the best studied model systems for chemotaxis - Dictyostelium discoideum. ^ It was initially found that comparing to wild type cells, gsk3 - cells showed aberrant chemotaxis with a significant decrease in both speed and chemotactic indices. In Dictyostelium, phosphatidylinositol 3,4,5-triphosphate (PIP3) signaling is one of the best characterized pathways that regulate chemotaxis. Molecular analysis uncovered that gsk3- cells suffer from high basal level of PIP3, the product of PI3K. Upon chemoattractant cAMP stimulation, wild type cells displayed a transient increase in the level of PIP3. In contrast, gsk3- cells exhibited neither significant increase nor adaptation. On the other hand, no aberrant dynamic of phosphatase and tensin homolog (PTEN), which antagonizes PI3K function, was observed. Upon membrane localization of PI3K, PI3K become activated by Ras, which will in turn further facilitate membrane localization of PI3K in an F-Actin dependent manner. The gsk3- cells treated with F-Actin inhibitor Latrunculin-A showed no significant difference in the PIP3 level. ^ I also showed GSK3 affected the phosphorylation level of the localization domain of PI3K1 (PI3K1-LD). PI3K1-LD proteins from gsk3- cells displayed less phosphorylation on serine residues compared to that from wild type cells. When the potential GSK3 phosphorylation sites of PI3K1-LD were substituted with aspartic acids (Phosphomimetic substitution), its membrane localization was suppressed in gsk3- cells. When these serine residues of PI3K1-LD were substituted with alanine, aberrantly high level of membrane localization of the PI3K1-LD was monitored in wild type cells. Wild type, phosphomimetic, and alanine substitution of PI3K1-LD fused with GFP proteins also displayed identical localization behavior as suggested by the cell fraction studies. Lastly, I identified that all three potential GSK3 phosphorylation sites on PI3K1-LD could be phosphorylated in vitro by GSK3.^

Compilation of respiration rate data for marine pelagic organisms

The metabolic rate of organisms may either be viewed as a basic property from which other vital rates and many ecological patterns emerge and that follows a universal allometric mass scaling law; or it may be considered a property of the organism that emerges as a result of the organism's adaptation to the environment, with consequently less universal mass scaling properties. Data on body mass, maximum ingestion and clearance rates, respiration rates and maximum growth rates of animals living in the ocean epipelagic were compiled from the literature, mainly from original papers but also from previous compilations by other authors. Data were read from tables or digitized from graphs. Only measurements made on individuals of know size, or groups of individuals of similar and known size were included. We show that clearance and respiration rates have life-form-dependent allometries that have similar scaling but different elevations, such that the mass-specific rates converge on a rather narrow size-independent range. In contrast, ingestion and growth rates follow a near-universal taxa-independent ~3/4 mass scaling power law. We argue that the declining mass-specific clearance rates with size within taxa is related to the inherent decrease in feeding efficiency of any particular feeding mode. The transitions between feeding mode and simultaneous transitions in clearance and respiration rates may then represent adaptations to the food environment and be the result of the optimization of tradeoffs that allow sufficient feeding and growth rates to balance mortality.

Antarctic sympagic meiofauna in winter

This study of Antarctic sympagic meiofauna in pack ice during late winter compares communities between the perennially ice-covered western Weddell Sea and the seasonally ice-covered southern Indian Ocean. Sympagic meiofauna (proto- and metazoans > 20 µm) and eggs > 20 µm were studied in terms of diversity, abundance and carbon biomass, and with respect to vertical distribution. Metazoan meiofauna had significantly higher abundance and biomass in the western Weddell Sea (medians: 31.1 * 10**3/m**2 and 6.53 mg/m**2, respectively) than in the southern Indian Ocean (medians: 1.0 * 10**3 /m**2 and 0.06 mg/m**2, respectively). Metazoan diversity was also significantly higher in the western Weddell Sea. Furthermore, the two regions differed significantly in terms of meiofauna community composition, as revealed through multivariate analyses. The overall diversity of sympagic meiofauna was high, and integrated abundance and biomass of total meiofauna were also high in both regions (0.6 - 178.6 * 10**3/m**2 and 0.02 - 89.70 mg/m**2, respectively), mostly exceeding values reported earlier from the western Weddell Sea in winter. We attribute the differences in meiofauna communities between the two regions to the older first-year ice and multi-year ice that is present in the western Weddell Sea, but not in the southern Indian Ocean. Our study indicates the significance of perennially ice-covered regions for the establishment of diverse and abundant meiofauna communities. Furthermore, it highlights the potential importance of sympagic meiofauna for the organic matter pool and trophic interactions in sea ice.

Pigments of sediment cores of the Northwest European Continental Margin

In the context of the European OMEX Programme this investigation focused on gradients in the biomass and activity of the small benthic size spectrum along a transect across the Goban Spur from the outer Celtic Sea into Porcupine Abyssal Plain. The effects of food pulses (seasonal, episodic) on this part of the benthic size spectrum were investigated. Sediments sampled during eight expeditions at different seasons covering a range from 200 m to 4800 m water depth were assayed with biochemical bulk measurements: determinations of chloroplastic pigment equivalents (CPE), the sum of chlorophyll a and its breakdown products, provide information concerning the input of phytodetrital matter to the seafloor; phospholipids were analyzed to estimate the total biomass of small benthic organisms (including bacteria, fungi, flagellata, protozoa and small metazoan meiofauna). A new term 'small size class biomass' (SSCB) is introduced for the biomass of the smallest size classes of sediment-inhabiting organisms; the reduction of fluorescein-di-acetate (FDA) was determined to evaluate the potential activity of ester-cleaving bacterial exoenzymes in the sediment samples. At all stations benthic biomass was predominantly composed of the small size spectrum (90% on the shelf; 97-98% in the bathyal and abyssal parts of the transect). Small size class biomass (integrated over a 10 cm sediment column) ranged from 8 g C/m**2 on the shelf to 2.1 g C/m**2 on the adjacent Porcupine Abyssal Plain, exponentially decreasing with increasing water depth. However, a correlation between water depth and SSCB, macrofauna biomass as well as metazoan meiofauna biomass exhibited a significantly flatter slope for the small size classes in comparison to the larger organisms. CPE values indicated a pronounced seasonal cycle on the shelf and upper slope with twin peaks of phytodetrital deposition in mid spring and late summer. The deeper stations seem to receive a single annual flux maximum in late summer. SSCB and heterotrophic activity are significantly correlated to the amount of sediment-bound pigments. Seasonality in pigment concentrations is clearly followed by SSCB and activity. In contrast to macro- and megafauna which integrate over larger periods (months/years), the small benthic size classes, namely bacteria and foraminifera, proved to be the most reactive potential of the benthic communities to any perturbations on short time scales (days/weeks). The small size classes, therefore, occupy a key role in early diagenetic processes.

Fossil associations from the middle and upper Eocene strata of the Pamplona Basin and surrounding areas (Navarre, western Pyrenees)

Fossil associations from the middle and upper Eocene (Bartonian and Priabonian) sedimentary succession of the Pamplona Basin are described. This succession was accumulated in the western part of the South Pyrenean peripheral foreland basin and extends from deep-marine turbiditic (Ezkaba Sandstone Formation) to deltaic (Pamplona Marl, Ardanatz Sandstone and Ilundain Marl formations) and marginal marine deposits (Gendulain Formation). The micropalaeontological content is high. It is dominated by foraminifera, and common ostracods and other microfossils are also present. The fossil ichnoasssemblages include at least 23 ichnogenera and 28 ichnospecies indicative of Nereites, Cruziana, Glossifungites and ?Scoyenia-Mermia ichnofacies. Body macrofossils of 78 taxa corresponding to macroforaminifera, sponges, corals, bryozoans, brachiopods, annelids, molluscs, arthropods, echinoderms and vertebrates have been identified. Both the number of ichnotaxa and of species (e. g. bryozoans, molluscs and condrichthyans) may be considerably higher. Body fossil assemblages are comparable to those from the Eocene of the Nord Pyrenean area (Basque Coast), and also to those from the Eocene of the west-central and eastern part of South Pyrenean area (Aragon and Catalonia). At the European scale, the molluscs assemblages seem endemic from the Pyrenean area, although several Tethyan (Italy and Alps) and Northern elements (Paris basin and Normandy) have been recorded. Palaeontological data of studied sedimentary units fit well with the shallowing process that throughout the middle and late Eocene occurs in the area, according to the sedimentological and stratigraphical data.

LtpD is a novel legionella pneumophila effector that binds phosphatidylinositol 3-phosphate and inositol monophosphatase IMPA1

The Dot/Icm type IV secretion system (T4SS) of Legionella pneumophila is crucial for the pathogen to survive in protozoa and cause human disease. Although more than 275 effector proteins are delivered into the host cell by the T4SS, the function of the majority is unknown. Here we have characterized the Dot/Icm effector LtpD. During infection, LtpD localized to the cytoplasmic face of the membrane of the Legionella-containing vacuole (LCV). In A549 lung epithelial cells, ectopically expressed LtpD localized to large vesicular structures that contained markers of endosomal compartments. Systematic analysis of LtpD fragments identified an internal 17-kDa fragment, LtpD_471-626, which was essential for targeting ectopically expressed LtpD to vesicular structures and for the association of translocated LtpD with the LCV. LtpD_471-626 bound directly to phosphatidylinositol 3-phosphate [PtdIns(3)P] in vitro and colocalized with the PtdIns(3)P markers FYVE and SetA in cotransfected cells. LtpD was also found to bind the host cell enzyme inositol (myo)-1 (or 4)-monophosphatase 1, an important phosphatase involved in phosphoinositide production. Analysis of the role of LtpD in infection showed that LtpD is involved in bacterial replication in THP-1 macrophages, the larvae of Galleria mellonella, and mouse lungs. Together, these data suggest that LtpD is a novel phosphoinositide- binding L. pneumophila effector that has a role in intracellular bacterial replication. © 2013, American Society for Microbiology.

Nueva especie de microalga y su aplicación para consumo animal, humano y en la obtención de carotenoides

Una nueva cepa de Scenedesmus ha sido aislada en la estación experimental "Las Palmerillas-CAJAMAR", siendo identificada como "microorganismo no previamente registrado" por la Universidad de Gottinghem, y depositada en la Culture Collection of Alagae and Protozoa (CCAP) como Scenedesmus almeriensis. La nueva cepa puede ser utilizada para consumo animal y/o humano, y produce elevadas cantidades de carotenoides, especialmente luteína y betacaroteno. Scenedesmus almeriensis crece adecuadamente en un amplio rango de temperaturas de 10°C a 40°C, a un pH de 7.0 a 9.5, y es tolerante a elevadas concentraciones de cobre, de hasta 1 mg/L. La cepa microalgal, cultivada en un fotobiorreactor de 4000 L en el que el cultivo se impulsa mecánicamente, produce luteína en una cantidad de al menos 4 mg por gramo de materia seca. Esta cepa es muy adecuada para la producción de carotenoides de aplicación en el tratamiento de desordenes de la macular ocular.

O microbioma humano

O organismo humano encontra-se colonizado por uma complexa diversidade de microrganismos. O conjunto destes microrganismos, que incluem as bactérias, fungos, vírus e protozoários, no homem denomina- se microbioma humano. Estima-se que entre a superfície interna e externa, o microbioma humano seja composto por 100 triliões de microrganismos. O microbioma humano varia muito nas mais diversas regiões do nosso corpo, dependendo de condições ambientais. Sabe-se, por exemplo, que nas regiões mais húmidas e quentes encontram-se uma maior concentração de microrganismos, enquanto que nas regiões menos húmidas, existe uma quantidade menor de microrganismos. O microbioma é de vital importância para a saúde humana, e o seu estudo conduz a um melhor conhecimento da sua complexa dinâmica, podendo conduzir ao desenvolvimento de novas formas de diagnóstico e até mesmo de tratamento de certas patologias. Assim sendo, a compreensão da diversidade fisiológica humana, bem como a de outros animais, passa pelo conhecimento da distribuição destes microrganismos nos diferentes órgãos e seu papel biológico. O desenvolvimento de técnicas de genética permitiram o estudo metagenómico importante para descrever a diversidade do microbioma humano, que não seria possível através da cultura das espécies pois um grande número destas não é cultivável. Esta dissertação tem como principal objetivo descrever o microbioma humano e de que forma esta influência o sistema imunitário. E numa segunda parte dar uma visão sobre a importância da análise metagenómica na identificação e caraterização do microbioma Humano.

Bacterioplâncton de quatro lagos rasos subtropicais com distintas características tróficas

O papel exercido pelas bactérias é reconhecido como fundamental no metabolismo de qualquer sistema aquático, não só pela mineralização da matéria orgânica, como também pela transferência de matéria e energia para níveis tróficos superiores (“microbial loop”). Para a realização deste estudo foram escolhidos quatro lagos com diferentes estados tróficos no Campus Carreiros da Universidade Federal do Rio Grande – FURG - RS. O Lago Biguás e o da Base possuem características de ambientes eutrófico - hipereutrófico, enquanto que, o Lago Polegar é caracterizado como um ambiente oligo-mesotrófico e o Lago Negro é considerado um ambiente distrófico. Em um estudo anterior em nove lagos rasos nesta mesma região, incluindo os quatro analisados no presente trabalho, Souza (2007) sugeriu que as bactérias livres atuariam como mineralizadoras e o seu crescimento seria limitado pela disponibilidade de fosfato (controle “bottom-up”), enquanto que as bactérias aderidas participariam da decomposição dos agregados orgânicos. Também foi sugerido que as bactérias aderidas seriam controladas principalmente pela predação por flagelados e ciliados (controle “top-down”), provavelmente devido ao seu maior biovolume. Porém, estas informações foram obtidas a partir de relações estatísticas de dados coletados em uma única amostragem. Assim, neste estudo a comunidade bacteriana (abundância e biomassa) e outros parâmetros físicos, químicos e biológicos dos quatro lagos rasos sub-tropicais foram estudados em amostragens quinzenais no decorrer de um ano entre junho de 2008 e maio de 2009. Nossos resultados indicam que a disponibilidade de carbono orgânico dissolvido produzido pelo fitoplâncton parece ser um dos principais fatores controladores da dinâmica de bactérias nestes lagos. Entretanto, a predação no Lago Negro parece ter sido de maior magnitude no controle das bactérias neste ambiente, uma vez que não houve um incremento na abundância bacteriana deste lago proporcional ao incremento da clorofila a. A presença de um maior número de nano - e microflagelados neste lago dá suporte a esta hipótese. Para testar esta hipótese, foi realizado um experimento utilizando-se a Técnica da Diluição em conjunto com a técnica a de FISH (Hibridização in situ Fluorescente) para identificar as taxas de produção e consumo não só dos diferentes morfotipos, mas também dos diferentes grupos filogenéticos (Archaea, Eubacteria, Alfa- Beta- e Gama-Proteobacteria e Cytophaga-Flavobacter) de uma amostra de água do Lago Negro. Os resultados deste experimento indicaram que as bactérias estão, de fato, sendo consumidas por vi protozoários na mesma proporção que estão sendo produzidas. Além disso, no Lago Negro a predação parece estar vinculada ao tamanho/biovolume celular, sendo os morfotipos de tamanho reduzido mais resistentes a predação e, por isso, mais abundantes.

Preliminary plankton research in the Bay of Nhatrang, Vietnam

An account is given of the work done on plankton of the Bay of Nhatrang, beginning with a study made by Rose in 1926. At the present time identification studies are being conducted at the Faculty of Sciences in Saigon, bearing on diatoms and protozoa

Survey of parasitic infection of Sander lucioperca (Linnaeus, 1758) and Silurus glanis (Linnaeus, 1758) in Aras reservoir with emphasizes of monogenean parasites

The parasites fauna of 491 specimens of Sander lucioperca, Linnaeus 1758 (246 specimens) and catfish, Silurus glanis, Linnaeus 1758 (245 specimens) in different size from Aras Reservoir situated in North —west of the Iran was investigated. During 2006-2007 Totally 16 parasite species were recorded. The most various parasites was found in catfish (10 species) while the lowest number was recorded in Sander lucioperca (6 species). Among them, however three genera of protozoa (Trichodina, Vorticella, Ichthyophthirius), two genera of Monogenea (Gyrodactylus, Silurodiscoides), Digenea, Cestoda, Nematoda, Acanthocephala and Annelida one species each (Diplostomum, Protocephalus, Eustrongylides, Neoechinorhynchus, Pisicola) and two crustacean genera (Argulus and Lernea) recorded and we can come to conclusion in comparison with the earlier data the actual parasite fauna of two hosts has been greatly improved. According to the present study the prevalence, mean abundance and mean intensity of parasites species of both hosts were highly influenced by seasons of the year. Some species found, however show a tendency to be more abundant ides Trichodina sp., Ichthyophthirius multifiliis, Silurodiscoides vistolensis, Protocephalus osculatus respectively. Most parasites species live in gills and skin, where is highly sensitive to some pathogens parasites species (Trichodina, Vorticella, Ichthyophthirius, Pisicola geometra, Argulus foliaceus; Lernea) and While some are specialist (Silurodiscoides vistolensis and Silurodiscoides siluri) other more or less generalist (ichthyophthirius).

Uma Lectina D-lactose específica da esponja marinha Cinachyrella apion: purificação, caracterização e interação com Leishmania chagasi

Four different sponge species were screened using Ouchterlony agarose gel and immunodiffusion tests to identify cross-reactivity with the polyclonal antibody IgG anti-deglicosilated CvL, a lectin from Cliona varians. Crude extract from the sponge Cinachyrella apion showed cross-reactivity and also a strong haemmaglutinating activity towards human erythrocytes of all ABO groups. Thus, it was submitted to acetone fractionation, IgG anti-deglicosilated CvL Sepharose affinity chromatography, and Fast Protein Liquid Chromatography (FPLC-AKTA) gel filtration on a Superose 6 10 300 column to purify a novel lectin. C. apion lectin (CaL) agglutinated all types of human erythrocytes with preference for papainized type A and O erythrocytes. The haemagglutinating activity is independent of Ca2+, Mg2+ and Mn2+ ions, and it was strongly inhibited by the disaccharide D-lactose, up to a minimum concentration of 6.25 mM. CaL molecular mass determined by FPLC-AKTA gel filtration on a Superose 12 10 300 column and SDS gel electrophoresis was approximately 124 kDa, consisting of eight subunits of 15.5 kDa, assembled by hydrophobic interactions. The lectin was relatively heat- and pH-stable. Leishmania chagasi romastigotes were agglutinated by CaL, indicating that lactose receptors could be presented in this parasite stage. These findings are indicative of the physiological defense roles of CaL and its possible use in the antibiosis of pathogenic protozoa