Fine structure of Chloromyxum menticirrhi n. sp. (Myxozoa) infecting the urinary bladder of the marine teleost Menticirrhus americanus (Sciaenidae) in Southern Brazil

A myxosporidian was found in the urinary bladder of the teleost Menticirrhus americanus Linnaeus, 1758 (Sciaenidae) collected from the South Atlantic coast of Brazil. Polysporic amoeboid plasmodia containing sporoblasts, developing pansporoblasts and spores were free in the bladder lumen. The prevalence of infection was 17.64% (15/85). Unfixed spores were spherical to subspherical, on average 10.5 μm long, 9.8 μm wide and 10.1 μm thick (n=25), and fixed spores measured 10.1×9.5×9.7 μm. The two spore valves were of equal size and each possessed prominent sutural lines and about 41 (37–45) surface ridges aligned parallel with the suture line. These ridges gave transverse sections a cog-wheel-like outline. The spores contained four pyriform polar capsules of equal size (3.20×2.0 μm) (n=25) (fixed), each with a polar filament having 3–4 (rarely 5) coils. The binucleate sporoplasm was irregular in shape, with granular matrix and randomly distributed dense bodies. The shape and dimensions of the spore, as well as the number, position and arrangement of the surface ridges, polar capsules and polar filament indicate that this is a new species, herein designated Chloromyxum menticirrhi. The gill, liver, gall bladder and intestine of the host showed no abnormalities.     

Morphology and phylogeny of Henneguya jocu n. sp. (Myxosporea,Myxobolidae), infecting the gills of the marine fish Lutjanus jocu

Henneguya jocu n. sp. (Myxosporea, Myxobolidae) is described from the gill lamellae of the marine teleost fish Lutjanus jocu, with a focus on ultrastructural and molecular features. This myxosporean forms subspherical cysts up to ∼260 μm × 130 μm long, and develops asynchronously. Mature myxospores ellipsoidal with a bifurcated caudal process. Myxospore length 10.9 ± 0.4 μm (n = 50); width, 8.2 ± 0.3 μm (n = 50); and thickness, 2.9 ± 0.5 μm (n = 50). Two equal caudal processes, 34.1 ± 1.0 μm long (n = 50); and total myxospore length, 45.2 ± 1.0 μm (n = 50). Two symmetric valves surround two ellipsoidal polar capsules, 5.0 ± 0.3 × 1.4 ± 0.2 μm (n = 20), each containing an isofilar polar filament forming 4–5 coils along the inner wall of these structures, as well as a binucleated sporoplasm presenting a spherical vacuole and several globular sporoplasmosomes. Both the morphological data and molecular analysis of the SSU rDNA gene identify this parasite as a new species of the genus Henneguya. Maximum Likelihood and Maximum Parsimony analyses further indicate that the parasite clusters within others marine Myxobolidae species, forming a group alongside other Henneguya species described from marine hosts.     

Glomerella truncata sp. nov., the teleomorph of Colletotrichum truncatum

Anthracnose of lentil, caused by Colletotrichum truncatum is a serious threat to lentil (Lens culinaris) grown in western Canada. The teleomorph stage of this pathogen was induced to form under laboratory conditions. Random pairing of single conidium isolates enabled the identification of fertile isolates. The individual isolates of this fertile pair were crossed with 14 other isolates, and all isolates were also incubated alone. Self-sterility was observed for all 16 isolates tested. Three isolates did not produce perithecia with either tester isolate, and none of the isolates tested produced perithecia with both tester isolates. Perithecia were brown–black, superficial, solitary or in small groups, obpyriform to ovate or ampulliform, 200–520 × 110–320 μm (mean: 350 × 200 μm). Asci were cylindrical, narrowing slightly at the apex, unitunicate, evanescent, 53–142 × 5–14 μm (mean: 90 × 8 μm), and contained eight ascospores. Ascospores were hyaline, aseptate, oblong, 12–20 × 5–8 μm (mean: 15.7–6.7 μm). The characteristics agree with those described for the genus Glomerella, and the species was named G. truncata sp. nov. The morphology of the new species is compared with that of other species in the genus, and future research on G. truncata is described.     

The effects of light intensity on the growth of Japanese Gambierdiscus spp. (Dinophyceae)

Marine toxic dinoflagellates of the genus Gambierdiscus are the causative agents of ciguatera fish poisoning (CFP), a form of seafood poisoning that is widespread in tropical, subtropical and temperate regions worldwide. The distributions of Gambierdiscus australes, Gambierdiscus scabrosus and two phylotypes of Gambierdiscus spp. type 2 and type 3 have been reported for the waters surrounding the main island of Japan. To explore the bloom dynamics and the vertical distribution of these Japanese species and phylotypes of Gambierdiscus, the effects of light intensity on their growth were tested, using a photoirradiation-culture system. The relationship between the observed growth rates and light intensity conditions for the four species/phylotypes were formulated at R > 0.92 (p < 0.01) using regression analysis and photosynthesis-light intensity (P-L) model. Based on this equation, the optimum light intensity (L_max) and the semi-optimum light intensity range (L_s-opt) that resulted in the maximum growth rate (μ_max) and ≥80% μ _max values of the four species/phylotypes, respectively, were as follows: (1) the L_max and L_s-opt of G. australes were 208 μmol photons m⁻² s⁻¹ and 91–422 μmol photons m⁻² s⁻¹, respectively; (2) those of G. scabrosus were 252 and 120–421 μmol photons m⁻² s⁻¹, respectively; (3) those of Gambierdiscus sp. type 2 were 192 and 75–430 μmol photons m⁻² s⁻¹, respectively; and (4) those of Gambierdiscus sp. type 3 were ≥427 and 73–427 μmol photons m⁻² s⁻¹, respectively. All four Gambierdiscus species/phylotypes required approximately 10 μmol photons m⁻² s⁻¹ to maintain growth. The light intensities in coastal waters at a site in Tosa Bay were measured vertically at 1 m intervals once per season. The relationships between the observed light intensity and depth were formulated using Beer’s Law. Based on these equations, the range of the attenuation coefficients at Tosa Bay site was determined to be 0.058–0.119 m⁻¹. The values 1700 μmol photons m⁻² s⁻¹, 500 μmol photons m⁻² s⁻¹, and 200 μmol photons m⁻² s⁻¹ were substituted into the equations to estimate the vertical profiles of light intensity at sunny midday, cloudy midday and rainy midday, respectively. Based on the regression equations coupled with the empirically determined attenuation coefficients for each of the four seasons, the ranges of the projected depths of L_max and L_s-opt for the four Gambierdiscus species/phylotypes under sunny midday conditions, cloudy midday conditions, and rainy midday conditions were 12–38 m and 12–54 m, 1–16 m and 1–33 m, and 0 m and 0–16 m, respectively. These results suggest that light intensity plays an important role in the bloom dynamics and vertical distribution of Gambierdiscus species/phylotypes in Japanese coastal waters.     

A new record of Myxobolus brachysporus and M. israelensis in the tilapia (Oreochromis niloticus) collected from the Nile River,Egypt

The present study was carried out as part of an ongoing general survey for myxosporean parasites infecting tilapias in the River Nile, Egypt. In the present study, 77 Nile tilapia (Oreochromis niloticus) were collected from boat landing sites at Beni-Suef governorate, Egypt and examined for the myxosporean infection. The infection was encountered as a huge number of free spores in the kidney and the spleen. The infection showed a prevalence of 51.9% (40/77) for Myxobolus brachysporus while it was 25.9% (20/77) for Myxobolus israelensis. Mature spores of M. brachysporus were ellipsoidal and measured 8.6 × 13.2 μm. The polar capsules were subcircular with 5–6 filament turns and measured 4.7 × 3.6 μm. Spores of M. israelensis were ellipsoidal in the frontal view and fusiform in the lateral view. Spore measurements were 13.4 μm long and 8.7 μm wide. The polar capsules were elongated with 6–7 filament coils and measured 8.6 × 3.1 μm. The findings presented here proved that tilapia fishes in the Nile River are still suffering from infections with Myxobolus species. Therefore, further studies should be carried out to survey the Myxobolus infection among tilapias under culture conditions to clarify the pathological impacts of this parasite in tilapias aquaculture.     

New antimicrobial anthraquinone 6,61-bis (1,5,7-trihydroxy-3-hydroxymethylanthraquinone) isolated from Streptomyces sp. isolate ERI-26

The present report is about Streptomyces sp. isolate ERI-26 isolated from the soil sample of Nilgiri forest, Western Ghats. The methanol extract of ERI-26 showed good antimicrobial activity against tested microbes. The antimicrobial novel anthraquinones were purified by bioactivity-guided fractionation using a silica gel column and preparative HPLC. The compound was characterized and identified by UV, IR, NMR and MASS spectral data. The compound named as 6,6¹-bis (1,5,7-trihydroxy-3-hydroxymethylanthraquinone), showed significant antimicrobial activities against tested microbes. The isolated compound inhibited the tested bacterial growth, Staphylococcus aureus at 62.5 μg/ml, Staphylococcus epidermidis at 15.62 μg/m, Bacillus subtilis at 62.5 μg/ml, fungi; Trichophyton mentagrophytes at 15.62 μg/m Trichophyton simii at 15.62 μg/ml, Aspergillus niger at. 7.81 μg/ml, Aspergiller flavus at 3.90 μg/ml, Trichophyton rubrum 296 at 62.5 μg/ml, T. rubrum 57/01 at 7.81 μg/ml, Magnaporthe grisea at 15.62 μg/ml. and Botrytis cinerea at 3.90 μg/ml. Isolated anthraquinone compound and its antimicrobial activity were newly reported.     

Changes in the size distribution of goat steroidogenic luteal cells during pregnancy

Experiments were conducted to investigate the size distribution of goat steroidogenic luteal cells throughout pregnancy. Corpora lutea were collected from very early (<6 weeks), early (6–8 weeks), middle (9–14 weeks) or late (15–18 weeks) stages of pregnancy. Luteal tissue was dissociated into single-cell suspension by enzyme treatments. Cells were stained for 3β-hydroxysteroid dehydrogenase (3β-HSD) activity, a marker for steroidogenic cells. The steroidogenic cells covered a wide spectrum of size ranging from 5 to 45 μm in diameter. There was a significant increase in mean cell diameter (P>0.01) as pregnancy progressed. Mean diameter of 3β-HSD positive cells increased from 14.73±0.35 μm in the corpus luteum of very early pregnancy to 24.20±0.45 μm in the corpus luteum of late pregnancy. The ratio of large (>20 μm in diameter) to small (5–20 μm in diameter) luteal cells was 0.28:1.0 in very early pregnancy, with the 7.5–15 μm cell size class being dominant. However, the ratio of large-to-small luteal cells was increased to 1.77:1.0 μm as pregnancy advanced and 25–35 μm cell sizes became predominant. It is likely that small luteal cells could develop into large cells as pregnancy progresses. Development of pregnancy is also associated with an increase in size of steroidogenic luteal cells.     

Nutritional effects of culture media on mycoplasma cell size and removal by filtration

Careful media filtration prior to use is an important part of a mycoplasma contamination prevention program. This study was conducted to increase our knowledge of factors that influence efficient filtration of mycoplasma. The cell size of Acholeplasma laidlawii was measured after culture in various nutritional conditions using scanning electron microscopy. The maximum cell size changed, but the minimum cell size remained virtually unchanged and all tested nutritional conditions resulted in a population of cells smaller than 0.2 μm. Culture in Tryptic Soy Broth (TSB) resulted in an apparent increase in the percentage of very small cells which was not reflected in increased penetration of non-retentive 0.2 μm rated filters. A. laidlawii cultured in selected media formulations was used to challenge 0.2 μm rated filters using mycoplasma broth base as the carrier fluid. We used 0.2 μm rated filters as an analytical tool because A. laidlawii is known to penetrate 0.2 μm filters and the degrees of penetration can be compared. Culture of A. laidlawii in TSB resulted in cells that did not penetrate 0.2 μm rated filters to the same degree as cells cultured in other media such as mycoplasma broth or in TSB supplemented with 10% horse serum.     

Exogenous and endogenous stages of Eimeria perforans naturally infected domestic rabbit (Oryctolagus cuniculus) in Saudi Arabia: Light microscopic study

Exogenous and endogenous stages of Eimeria perforans naturally infected rabbits in Saudi Arabia were described. The prevalence of infection was 75%. Oocysts were ovoid to elliptical and measured 16 × 10 μm. The four dizoic sporocysts were ovoid and measured 7 × 5 μm. Endogenous stages were restricted to the duodenum. Meronts, microgamonts, macrogamonts and young oocysts were recorded and described.     

Crocodylus niloticus (Crocodilia) is highly sensitive to water surface waves

Crocodiles show oriented responses to water surface wave stimuli but up to now behavioral thresholds are missing. This study determines the behavioral thresholds of crocodilians to water surface waves. Nile crocodiles (Crocodylus niloticus) were conditioned to respond to single-frequency water surface wave stimuli (duration 1150 ms, frequency 15, 30, 40, 60 and 80 Hz), produced by blowing air onto the water surface. Our study shows that C. niloticus is highly sensitive to capillary water surface waves. Threshold values decreased with increasing frequency and ranged between 10.3 μm (15 Hz) and 0.5 μm (80 Hz) peak-to-peak wave amplitude. For the frequencies 15 Hz and 30 Hz the sensitivity of one spectacled caiman (Caiman crocodilus) to water surface waves was also tested. Threshold values were 12.8 μm (15 Hz) down to 1.76 μm (30 Hz), i.e. close to the threshold values of C. niloticus. The surface wave sensitivity of crocodiles is similar to the surface wave sensitivity of semi-aquatic insects and fishing spiders but does not match the sensitivity of surface-feeding fishes which is higher by one to two orders of magnitude.     

p90 ribosomal S6 kinase 1 (RSK1) isoenzyme specifically regulates cytokinesis progression

The p90 ribosomal S6 kinase family (RSK1–4) of Ser/Thr kinases is a downstream component of the Ras-MAPK cascade responsible for regulating various cellular processes. Here, we examined the potential involvement of RSKs in regulating mitosis by transfecting HeLa cells with siRNAs targeting RSK1 and -2, which are the major isoforms. Depletion of RSK1 but not RSK2 triggered a significant accumulation of binucleated cells compared to control cells (0.5% vs. 10.5%, respectively); this was rescued by expression of exogenous RSK1 but not a kinase-defective mutant. Monitoring of cell division by time-lapse imaging revealed that the observed binucleation mainly stemmed from a failure to form and ingress the cleavage furrow during early cytokinesis. Immunocytochemical analysis of RhoA and anillin, the two principal regulators of cleavage furrow formation and ingression, showed that these proteins were abnormally localized during anaphase in RSK1-depleted cells. Furthermore, RSK1-depleted cells seemed to have impairments in midzone microtubule formation, as suggested by morphological changes and lengthening of the midzone (15.2 ± 1.7 μm vs. 17.4 ± 1.7 μm in control cells). We also observed shortening of the pole-to-polar-cortex distance in RSK1-depleted cells (4.30 ± 1.37 μm vs. 2.80 ± 0.84 μm in control cells) and scanty distribution of microtubules at the periphery of the equatorial region during anaphase, suggesting an aberrant distribution of astral microtubules. Taken together, these results suggest that RSK1 is specifically required for cleavage furrow formation and ingression during cytokinesis. This may occur via the involvement of RSK1 in proper midzone and astral microtubule structure formation during anaphase, which is essential for the correct localization of anillin and RhoA.     

Picrasin K,a new quassinoid from Quassia amara L. (Simaroubaceae)

A new quassinoid Picrasin K 1 was isolated from a decoction made of Quassia amara leaves, traditionally used in French Guyana to treat malaria. The structure and relative stereochemistry of 1 was determined through extensive NMR analysis. Picrasin K showed a low activity against Plasmodium falciparum in vitro (IC₅₀ = 8 μM), and a similar low activity on human cancerous cells line (IC₅₀ = 7 μM on MCF-7 cells line).     

Glomus drummondii and G. walkeri,two new species of arbuscular mycorrhizal fungi (Glomeromycota)

Two new ectocarpic arbuscular mycorrhizal fungal species, Glomus drummondii and G. walkeri (Glomeromycota), found in maritime sand dunes of northern Poland and those adjacent to the Mediterranean Sea are described and illustrated. Mature spores of G. drummondii are pastel yellow to maize yellow, globose to subglobose, (58–)71(–85) μm diam, or ovoid, 50–80 × 63–98 μm. Their wall consists of three layers: an evanescent, hyaline, short-lived outermost layer, a laminate, smooth, pastel yellow to maize yellow middle layer, and a flexible, smooth, hyaline innermost layer. Spores of G. walkeri are white to pale yellow, globose to subglobose, (55–)81(–95) μm diam, or ovoid, 60–90 × 75–115 μm, and have a spore wall composed of three layers: a semi-permanent, hyaline outermost layer, a laminate, smooth, white to pale yellow middle layer, and a flexible, smooth, hyaline innermost layer. In Melzer's reagent, only the inner- and outermost layers stain reddish white to greyish rose in G. drummondii and G. walkeri, respectively. Both species form vesicular–arbuscular mycorrhizae in one-species cultures with Plantago lanceolata as the host plant. Phylogenetic analyses of the ITS and parts of the LSU of the nrDNA of spores placed both species in Glomus Group B sensu Schüßler et al. [Schüßler A, Schwarzott D, Walker C, 2001. A new fungal phylum, the Glomeromycota: phylogeny and evolution. Mycolological Research 105: 1413-1421.]     

Ostreopsis cf. siamensis and Ostreopsis cf. ovata from the Atlantic Iberian Peninsula: Morphological and phylogenetic characterization

Individuals of Ostreopsis, a genus containing potentially toxic species which affects human health, were collected during summer-autumn 2010 and 2011 from 17 sites located along the Atlantic coast of the Iberian Peninsula, a temperate area which during summer presents contrasting seawater temperatures. Ostreopsis cells were obtained by shaking macroalgae collected from rocky-shore areas bordering accessible beaches. Isolated strains and field samples were analyzed for morphological and phylogenetic characterization where sequences of the ITS1-5.8S-ITS2 region of the rDNA delineated two different species fitting Ostreopsis cf. ovata and Ostreopsis cf. siamensis. By means of calcofluor staining and scanning electron microscopy, it was observed that field samples of both species exhibited a wide and overlapping range of dorsoventral as well as width values. Those cells presented 11–18 pores/100 μm² and were also similar concerning plates shape and size. The main differential feature between the two species was the presence of two sizes of thecal pores (0.07–0.13 μm and 0.15–0.39 μm) in Ostreopsis cf. siamensis and one size (0.24–0.56 μm) in Ostreopsis cf. ovata. A comparison of field vs. cultured cells indicated that field isolates presented larger cells than in culture.     

Pod,seed traits and cytotaxonomic studies of some Vicia narbonensis L. accessions (Fabaceae)

This study aimed to characterize eight accessions of Vicia narbonensis L. originated from different Mediterranean countries. The cytology of these species is rarely known despite the fact of its great socio-economical and ecological interest in these arid and semi-arid zones. This work aimed mainly to characterize the karyotype, morphological pod and seed traits of the species. Karyotypes of all accessions were similar to a diploid number of 2n = 2x = 14. All the accessions have submetacentric chromosomes with a secondary constriction attached to the long arm of pair VII close to the centromere. Variation in chromosome size was observed; it ranged from 5.86 μm to 7.62 μm. Indices of karyotype asymmetry were calculated as the total form percentage (TF%) and symmetric indices (Syi) which ranged from 33.75% to 35.42% and from 51.01% to 54.85%, respectively. The predominance of submetacentric chromosomes indicated that the karyotype is symmetrical and can be considered as primitive. However, the analysis of quantitative parameters measured on pods and seeds showed a significant variation between accessions. A relationship between centromeric index and the pod beak length was found. Estimation of phenotypic diversity using the Shannon diversity index (H′) showed that the length, the seed color and the number of seeds per pod are the most polymorphic traits with respectively, H′ = 0.92, 0.80 and 0.83. Cluster analysis of karyological, pod and seed traits showed four groups of accessions. This clustering is partially due to the geographical origin of the studied accessions. The variation in chromosome size, pod and seed traits could offer potentially valuable genetic resources for the improvement of V. narbonensis which is considered as neglected and underutilized crop species (NUCS).     

Green biosynthesis of silver nanoparticles from Annona squamosa leaf extract and its in vitro cytotoxic effect on MCF-7 cells

The biological method for the synthesis of silver nanoparticles (AgNPs) using Annona squamosa leaf extract and its cytotoxicity against MCF-7 cells are reported. The synthesized AgNPs using A. squamosa leaf extract was determined by UV–visible spectroscopy and it was further characterized by FT-IR, X-ray diffraction (XRD), Transmission electron microscopy (TEM), Zeta potential and energy dispersive spectrometric (EDS) analysis. The UV–visible spectrum showed an absorption peak at 444 nm which reflects surface plasmon resonance (SPR) of AgNPs. TEM photography showed biosynthesized AgNPs were predominantly spherical in shape with an average size ranging from 20 to 100 nm. The Zeta potential value of ?37 mV revealed the stability of biosynthesized AgNPs. Furthermore, the green synthesized AgNPs exhibited a dose-dependent cytotoxicity against human breast cancer cell (MCF-7) and normal breast epithelial cells (HBL-100) and the inhibitory concentration (IC₅₀) were found to be 50 μg/mL, 30 μg/mL, and 80 μg/mL, 60 μg/ml for AgNPs against MCF-7 and normal HBL-100 cells at 24 h and 48 h incubation respectively. An induction of apoptosis was evidenced by (AO/EtBr) and DAPI staining. Application of such eco-friendly nanoparticles makes this method potentially exciting for the large scale synthesis of nanoparticles.     

Biomorphometric characteristics of different types of sensilla detected on the antenna of Helicoverpa armigera by scanning electron microscopy

A morphometric study on H. armigera antenna showed four styles of sensilla, i.e., styloconica, chaetica, coeloconica, and trichodea, and their numbers were estimated. Sensilla trichodea detect inter and intraspecific communication signals and was the most numerous. They were divided into three types: type I, the longest, with a length of 34.04 ± 3.16 μm and about 2.16 to 2.42 μm in diameter at its base; 2) type II, intermediate, with a length of 22.58 ± 0.77 μm and basal diameter of 1.8–2.52 μm; 3) type III, the shortest sensilla trichodea, with a length of 7.62 ± 0.4 μm and a range in diameter similar to that of type II. The length of the female sensilla trichodea was longer than that of the male. The total number of sensilla trichodea was estimated to be 7520 on the antenna of the female, and 6831 on the male antenna. The lengths of the sensilla trichodea type I and type III were significantly different on male (t = 4.6881, P = 0.0034) and female antenna (t = 18.9852, P = 0.0001). An estimation of the predicted surface area of the most numerous type I on sampled segments between the 12th and 20th segments from a female of H. armigera showed a surface area of 5 × 10³ μm² and a sensillar density of 38 sensilla/10³ μm². The fraction of sensilla-occupied surface area was 0.4 μm².     

Serotonin transporter protein (SERT) and P-glycoprotein (P-gp) binding activity of montanine and coccinine from three species of Haemanthus L. (Amaryllidaceae)

The alkaloid rich extracts from an acid/base extraction of bulb material of Haemanthus coccineus L., H. montanus Baker and H. sanguineus Jacq. revealed that two montanine type Amaryllidaceae alkaloids, montanine (1) and coccinine (2) were the major alkaloid constituents. Together these two alkaloids constituted 88, 91 and 98% of the total alkaloid extract from each species respectively. GC–MS analysis revealed that H. coccineus and H. sanguineus had a relative abundance of coccinine (74 and 91% respectively) to montanine (14 and 7% respectively); whereas H. montanus had 20% coccinine and 71% montanine. The three extracts and two isolated alkaloids were evaluated for binding to the serotonin transporter protein (SERT) in vitro. Affinity to SERT was highest in H. coccineus (IC₅₀ = 2.0 ± 1.1 μg/ml) followed by H. montanus (IC₅₀ = 6.8 ± 1.0 μg/ml) and H. sanguineus (IC₅₀ = 28.7 ± 1.1 μg/ml). Montanine (IC₅₀ = 121.3 ± 3.6 μM or 36.56 ± 1.14 μg/ml; K_i = 66.01 μM) was more active than coccinine (IC₅₀ = 196.3 ± 3.8 μM or 59.15 ± 1.08 μg/ml; K_i = 106.8 μM), both of which were less active than the total alkaloid extracts of each species investigated. The possible synergistic effects of two coccinine/montanine mixtures (80:20 and 20:80) were investigated, however the mixtures gave similar activities as the pure compounds and did not show any increase in activity or activity similar to the total alkaloid extracts. Thus the considerably higher activity observed in the total alkaloid extracts is not correlated to the relative proportions of coccinine and montanine in the extracts and thus are likely to be due to more potent unidentified minor constituents. Both alkaloids exhibited low binding affinity to P-glycoprotein (P-gp) as demonstrated by low inhibition of calcein-AM efflux in the MDCK-MDR1 cell line. This indicates that P-gp efflux will not be limiting for blood–brain-barrier passage of the alkaloids.     

Reinvestigation of epithelial lining of the genital coelomic sinus in asteroids. An ultrastructural study

Ultrastructural study of gonadal muscles in sea star, Asterina pectinifera, showed that myoepithelial cells were located only in the epithelial lining of the genital coelomic sinus. No myoepithelial cells were found in the visceral peritoneal epithelium or within connective tissue layer of the outer sac. Morphology of the myoepithelial cells in gonads of A. pectinifera varies during the reproductive cycle. During the gametogenic phase of the reproductive cycle, the myoepithelial cells get an elongated, spindle-like shape having a length of 20–30 μm. In prespawning gonads, many of the myoepithelial cells form cytoplasmic extensions of 3–5 μm in length, filled with myofilaments and penetrating into the underlying connective tissue of the outer sac or haemal sinus. Besides, myoepithelial cells, simultaneously anchored in the inner and outer sacs, were also observed. These changes result in development of more elaborated musculature and increase in contractility of the gonadal wall in prespawning gonads as compared to that during other stages of the reproductive cycle.     

Morphology and bloom dynamics of Cochlodinium geminatum (Schütt) Schütt in the Pearl River Estuary,South China Sea

An unarmored dinoflagellate bloom of Cochlodinium geminatum (Schütt) Schütt has been identified in the Pearl River Estuary, South China Sea during the severe dry season, from late October to early November, 2009, when temperature and salinity ranged between 20.0–27.2 °C and 10.6–33.4, respectively. Light and scanning electron microscopy were used to identify the characteristics of C. geminatum and provided the clear morphological structure for this species. The organism was primarily found in chains of two cells or single cell, and no longer chains were observed. Cells were irregularly spherical or slightly dorso-ventrally, with size ranged between 28 and 36 μm and longer than wide. A large nucleus in the center with numerous golden chloroplasts was present, and the cingulum made 1.5 turns around the cell. The concentration of C. geminatum ranged from 10² to greater than 10⁷ cells l⁻¹ during the bloom period. Nutrient concentration ranges during the bloom were 1.29–81.00 μM NO₃, 0.14–12.14 μM NO₂, 0.21–6.29 μM NH₄, 0.23–6.26 μM PO₄ and 3.29–171.43 μM SiO₃, respectively. Total biomass expressed in terms of chlorophyll a ranged from 2.44 to 135.45 μg l⁻¹, with an average 19.9 μg l⁻¹ in surface water throughout the PRE. Two main clusters corresponding to the water sectors were defined with multivariate analysis (cluster and nMDS). Based on the composition and abundance of phytoplankton, spatial variations were observed at a significant level (ANOSIM, R = 0.44, P < 0.01). Although the pairwise correlation analysis detected no significant effect of any single environmental variable on the abundance of C. geminatum, the multivariate analysis (BIO-ENV) between biotic and abiotic variables resulted in the best variables combination with all measured factors involved (temperature, salinity, turbidity, NO₃, NO₂, NH₄, PO₄ and SiO₃) which showed a combined effect during the bloom of C. geminatum in the Pearl River Estuary (ρ_w = 0.477).