Types of collagen fibres in the calcareous sponges Sycon and Leucandra

Collagen from the calcareous sponges Sycon and Leucandra has been studied in thin sections and in sheaths from decalcified spicules. Sections show abundant thin fibrils 8–15 mμ in diameter and, in Sycon juveniles, 20 mμ fibrils with a clear 50 mμ period. The sheaths are also mainly composed of the thin fibrils but giant fibres up to 130 mμ in diameter and many μ in length are also found.     

A Lipidomic Approach to Understanding Free Fatty Acid Lipogenesis Derived from Dissolved Inorganic Carbon within Cnidarian-Dinoflagellate Symbiosis

The cnidarian-dinoflagellate symbiosis is arguably one of the most important within the marine environment in that it is integral to the formation of coral reefs. However, the regulatory processes that perpetuate this symbiosis remain unresolved. It is essential to understand these processes, if we are to elucidate the mechanisms that support growth and resource accumulation by coral host, and conversely, recently observed reduction and/or mortality of corals in response to rapid environmental change. This study specifically focused on one area of metabolic activity within the symbiosis, that of free fatty acid synthesis within both the dinoflagellate symbionts and cnidarian host. The main model system used was Aiptasia pulchella and Symbiodinium sp. in combination with aposymbiotic A. pulchella, the symbiotic coral Acropora millepora system and dinoflagellate culture. Fatty acids (FAs) were selected because of their multiple essential roles inclusive of energy storage (resource accumulation), membrane structure fluidity and cell signaling. The study addressed free FA lipogenesis by using a new method of enriched stable isotopic (¹³C) incorporation from dissolved inorganic carbon (DI¹³C) combined with HPLC-MS. FAs derived from DI¹³C aligned with a mixture of known lipogenesis pathways with the addition of some unusual FAs. After 120 hr, ¹³C-enriched FA synthesis rates were attributed to only a complex integration of both n–3 and n–6 lipogenesis pathways within the dinoflagellate symbionts. Furthermore, there was no detectible evidence of symbiont derived enriched isotope fatty acids, catabolized ¹³C derivatives or DI¹³C being directly utilized, in host late n–6 pathway long-chain FA lipogenesis. These findings do not align with a popular mutualistic translocation model with respect to the use of translocated symbiont photoassimilates in host long-chain FA lipogenesis, which has important connotations for linking nutrient sources with metabolite production and the dynamic regulation of this symbiosis.     

Sterols and related metabolites from five species of sponges

Free sterol fractions were isolated from the marine sponges Phyllospongia madagascarensis, Scalarispongia sp., Oceanapia sp., Monanchora clathrata and studied by GLC, GLC–MS, and spectroscopy NMR. P. madagascarensis and Scalarispongia sp. contained common Δ⁵-sterols; cholesterol was shown to be a main sterol of both the sponges. Oceanapia sp. contained stanols and minor Δ⁵-sterols with 24R-24,25-methylene-5α-cholestan-3β-ol as a main constituent. Many free sterols from M. clathrata were Δ⁷-series compounds, and latosterol was a main sterol. Δ⁴-3-Ketosteroids and Δ⁵-sterol esters were found in the Antarctic sponge Haliclona sp., but free sterols were practically absent except for trace amount of cholesterol. A chemotaxonomic application of sterols in relation to the genera Phyllospongia, Oceanapia and the family Crambeidae is provided. The known cases of the absence of sterols in sponges and probable reasons of the phenomenon are discussed.     

Seasonal variation in light-shade adaptation of natural populations of the symbiotic sea anemone Aiptasiapulchella (Carlgren, 1943) in Hawaii

The productivity and biomass parameters of the symbiotic anemone Aiptasia pulchella (Carlgren, 1943) from a shaded mangrove lagoon (maximum summer irradiance of 100 μE m⁻² · s⁻¹) and a sunlit reef flat (maximum summer irradiance of 1400 μE · m⁻² · s⁻¹) were examined in Hawaii. Light-shade adaptation was evident in the summer populations (1981) but not observed during the fall (1982). In the summer, zooxanthellae from the lagoon A. pulchella (shade anemones) contained 2.97 pg Chl a cell ⁻¹ and those from the reef flat (sun anemones) contained 1.70 pg Chl a · cell⁻¹; but Chl a : c₂ ratios were 2.5 in zooxanthellae from both shade and sun anemones. During the fall, there were no significant differences in Chl a and c₂ of zooxanthellae (2.25 pg Chl a · cell⁻¹) in shade and sun anemones, but Chl a : c₂ ratios averaged 3.9. During both seasons, shade anemones were larger and contained higher densities of zooxanthellae than sun anemones. In addition to differences between shade and sun habitats, there was localized photoadaptation of zooxanthellae within individual anemones due to microhabitat variations in ambient irradiance. Growth rates of zooxanthellae in A. pulchella differed in shade and sun anemones. Specific growth rates for zooxanthellae in situ were the same for shade populations in both summer and fall (0.016 day⁻¹). However, zooxanthellae in sun anemones grew four times faster in the fall (0.033 day⁻¹) than during the summer (0.008 day⁻¹). These results suggest that growth of zooxanthellae in these anemones was independent of ambient irradiance. Photosynthesis-irradiance (P-I) responses of shade and sun anemones during the summer showed that shade anemones had greater photosynthetic efficiencies (α) but lower photosynthetic capacities (P_max) than sun anemones. Dark-respiration rates of sun anemones were twice those obtained with shade anemones. In the fall, these populations of anemones did not exhibit P-I responses characteristic of light-shade adaptation. Both α and P_max of shade and sun anemones were higher in the fall, indicating that zooxanthellae in A. pulchella adapted to seasonal reduction in irradiance.     

A comparative study of the thermal stability of formate dehydrogenases from microorganisms and plants

A comparative study of the thermostability of NAD⁺-dependent formate dehydrogenases (FDHs; EC 1.2.1.2) from both methylotrophic bacteria Pseudomonas sp. 101 and Moraxella sp. C1, the methane-utilizing yeast Candida boidinii, and plants Arabidopsis thaliana and Glycine max (soybean) was performed. All the enzymes studied were produced by expression in E. coli cells. The enzymes were irreversibly inactivated in one stage according to first-order reaction kinetics. The FDH from Pseudomonas sp. 101 appeared as the most thermostable enzyme; its counterpart from Glycine max exhibited the lowest stability. The enzymes from Moraxella sp. C1, C. boidinii, and Arabidopsis thaliana showed similar thermostability profiles. The temperature dependence of the inactivation rate constant of A. thaliana FDH was studied. The data of differential scanning calorimetry was complied with the experimental results on the inactivation kinetics of these enzymes. Values of the melting heat were determined for all the enzymes studied.     

Flagellar apparatus structure of choanocyte in Sycon sp. and its significance for phylogeny of Porifera

The problem of the origin of Metazoa has been one of the most discussed for nearly the last one and a half centuries. Some 20 years ago, morphological approaches were replaced with molecules, but the problem then became more complex. At the same time, morphological data were incomplete, and therefore, this approach can still help in comparison with choanoflagellates and sponges—two sister groups having uniflagellated cells with a collar. The structure of the flagellar apparatus has phylogenetic significance, but sponge choanocytes are poorly studied in this respect, and we still do not know what the ancestral kinetid of Porifera looks like. The kinetid structure of choanocytes in Sycon sp. is investigated here for the first time, and a 3D reconstruction of the kinetid provided. It is composed of a flagellar kinetosome with a nuclear fibrillar root and a basal foot with a few microtubules; the accessory centriole lies orthogonal to and just below the foot of the kinetosome, and a dictyosome is near the centriole. This kinetid is similar to that of the choanocyte of Corticium candelabrum (Homoscleromorpha) and is considered to be the ancestral type for the whole branch Calcarea + Homoscleromorpha.     

Characterization of archaeal symbionts of sponges from the coral reef ecosystems of the Gulf of Mannar,Southeast coast of India

Sponges accommodate a diverse group of microorganisms with varied metabolic capabilities. The bacterial associates of sponges are widely studied while our understanding of archaeal counterparts is scanty. In the present study, we report the archaeal associates of two sponges, Pseudoceratina purpurea (NCBI barcode: KX454492) and Cinachyra sp. (NCBI barcode: KX454495), found in the coral reef ecosystems of Gulf of Mannar, India. Archaea in the water column was predominated by members of class Halobacteria of Phylum Euryarchaeota (97%) followed by a minor fraction (3%) of Nitrosopumilus sp. of phylum Thaumarchaeota. Interestingly, Thaumarchaeota was identified as the sole archaeal population associated with the two sponges studied, among which Nitrosopumilus sp. occuppied 80 and 100% of the sequences in the clone library of P. purpurea and Cinachyra sp. respectively. Other archaea found in the P. purpurea were Nitrososphaera (10%) and unclassified ones (10%). The study identified Nitrosopumilus sp. as a unique symbiotic archaeon of sponges, P. purpurea and Cinachyra sp. The existence of host driven factors in selecting specific associates from a diverse group of archaea in the environment may need further investigations.     

The energetics of asexual reproduction: Pedal laceration in the symbiotic sea anemone Aiptasia pulchella (Carlgren, 1943)

Asexual reproduction of the symbiotic sea anemone Aiptasia pulchella (Carlgren, 1943) was studied in the context of energy budgets determined under a variety of light and feeding conditions. Continuous darkness significantly increased rates of pedal laceration (the method of asexual reproduction), while different feeding rates had no significant effect. Digestive assimilation efficiency averaged 63% for whole brine shrimp, and conversion efficiencies (assimilated energy into biomass) averaged 31%. There were no significant differences between the conversion efficiencies of energy obtained from light or food. The index of reproductive effort (RE) for pedal laceration was very low, ranging from 0.004 to 0.044. This low RE associated with a substantial rate of asexual reproduction may, in part, explain the widespread success of certain sea anemones.     

Production of extracellular enzymes and degradation of biopolymers by saprotrophic microfungi from the upper layers of forest soil

Production of extracellular enzymes participating in the degradation of biopolymers was studied in 29 strains of nonbasidiomycetous microfungi isolated from Quercus petraea forest soil based on the frequency of occurrence. Most of the isolates were ascomycetes and belonged to the genera Acremonium, Alternaria, Cladosporium, Geomyces, Hypocrea, Myrothecium, Ochrocladosporium, and Penicillium (18 isolates), and two isolates were zygomycetes. Only six isolates showed phenol oxidation activity which was low and none of the strains were able to degrade humic acids. Approximately half of the strains were able to degrade cellulose and all but six degraded chitin. Most strains produced significant amounts of the cellulolytic enzymes cellobiohydrolase and ??-glucosidase and the chitinolytic enzymes chitinase, chitobiosidase, and N-acetylglucosaminidase. The highest cellulase activities were found in Penicillium strains, and the highest activity of chitinolytic enzymes was found in Acremonium sp. The production of the hemicellulose-degrading enzymes ??-galactosidase, ??-galactosidase, and ??-mannosidase was mostly low. The microfungal strains were able to produce significant growth on a range of 41?C87, out of 95 simple C-containing substrates tested in a Biolog? assay, monosaccharides being for all strains the most rapidly metabolized C-sources. Comparison with saprotrophic basidiomycetes from the same environment showed that microfungi have similar cellulolytic capabilities and higher chitinase activities which testifies for their active role in the decomposition of both lignocellulose and dead fungal biomass, important pools of soil carbon.     

Effects of free amino acids on the photosynthetic carbon metabolism of symbiotic dinoflagellates

Synthetic host factor (SHF) was used in parallel with crude cnidarian’s host factor (CHF) to investigate their effects on both photosynthetic carbon metabolism and the first step of lipid synthesis in the symbiotic dinoflagellates Symbiodium purchrorum of the sea anemone Aiptasia pulchella. Several species have been studied, namely, sea anemone A. pulchella, reef coral Pocillopora damicornis, and green alga Chlamydomonas reinhardii. Both short-term and long-term experiments with radioactive carbon have shown a higher rate of the alga ¹⁴C photoaccumulation with host factor(s) than of dinoflagellates located in artificial sea water (ASW) alone. In dinoflagellates incubated with both ASW and CHF, ¹⁴C-labeled glycerol was detectable after 15 s of alga illumination. In dinoflagellates isolated from P. damicornis and incubated in CHF and SHF and in dinoflagellates isolated from A. pulchella and incubated in CHF, a higher percentage of ¹⁴C was found in the glycerol as compared to the ASW trial. At the same time, in ASW trial the radioactive label was primarily located in ethanol-soluble lipid fraction. Similar results were observed when dinoflagellates isolated from P. damicornis were incubated with aspartate or glutamate. But there was no effect with taurine, serine, valine, glycine, or lysine. C. reinhardii, incubated in salt-free CHF, partitioned a greater percent of ¹⁴C into the glycerol and less into the ethanol-soluble lipids as compared to the corresponding control incubations. The amount of ¹⁴C in neutral and polar lipids was identical in that in A. pulchella dinoflagellates incubated in ASW or CHF. The arrays of neutral ¹⁴C-lipids produced under both ASW alone and ASW with CHF conditions, and over time were not significantly distinguishable. Host factors appeared to provide an optimum environment to sustain maximum metabolic efficiency. A biochemical model, based on the quantitative and qualitative assessment of carbon pathways in dinoflagellates incubated both in host factor and sea water alone, is presented. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 2, pp. 195–208. The text was submitted by the authors in English.     

Kinetic of the production of polygalacturonase and pectin lyase by two closely relatedFormae speciales ofFusarium oxysporum

The kinetic of thein vitro production of polygalacturonase and pectin lyase of two closely related fungi,Fusarium oxysporum f.sp.lycopersici andF. oxysporum f.sp.radicis-lycopersici, was examined under various culture conditions such as the source of carbon, the pH, and the age of cultures. Over a 5-day period, the production of these enzymes by various isolates of the sameforma specialis (f. sp.) ofF. oxysporum was not significantly different (P ≥ 0.05). However, the amount of the enzymes produced differed markedly between both f. sp. The different carbon sources added to the culture media, such as citrus pectin, apple pectin, tomato cell wall fragments, andd-galacturonic acid, proved to be higher pectinase inducible substrates than sucrose and glucose. For both fungi, polygalacturonase and pectin lyase activities were optimal at pH 5.0 and 8.0, respectively. Furthermore, pectin lyase production had a partial Ca²⁺ requirement in contrast to polygalacturonase production which was limited by Ca²⁺. In most experiments performed, the production of polygalacturonase appeared superior withF. oxysporum f.sp.radicislycopersici than withF. oxysporum f.sp.lycopersici. On the other hand, pectin lyase production ofF. oxysporum f.sp.lycopersici was approximately 10-fold greater than that byF. oxysporum f.sp.radicis-lycopersici in media supplemented withd-galacturonic acid.     

Biodiversity of Actinomycetes Associated with Caribbean Sponges and Their Potential for Natural Product Discovery

Marine actinomycetes provide a rich source of structurally unique and bioactive secondary metabolites. Numerous genera of marine actinomycetes have been isolated from marine sediments as well as several sponge species. In this study, 16 different species of Caribbean sponges were collected from four different locations in the coastal waters off Puerto Rico in order to examine diversity and bioactive metabolite production of marine actinomycetes in Caribbean sponges. Sediments were also collected from each location, in order to compare actinomycete communities between these two types of samples. A total of 180 actinomycetes were isolated and identified based on 16S rRNA gene analysis. Phylogenetic analysis revealed the presence of at least 14 new phylotypes belonging to the genera Micromonospora, Verruscosispora, Streptomyces, Salinospora, Solwaraspora, Microbacterium and Cellulosimicrobium. Seventy-eight of the isolates (19 from sediments and 59 from sponges) shared 100 % sequence identity with Micromonospora sp. R1. Despite having identical 16S rRNA sequences, the bioactivity of extracts and subsequent fractions generated from the fermentation of both sponge- and sediment-derived isolates identical to Micromonospora sp. R1 varied greatly, with a marked increase in antibiotic metabolite production in those isolates derived from sponges. These results indicate that the chemical profiles of isolates with high 16S rRNA sequence homology to known strains can be diverse and dependent on the source of isolation. In addition, seven previously reported dihydroquinones produced by five different Streptomyces strains have been purified and characterized from one Streptomyces sp. strain isolated in this study from the Caribbean sponge Agelas sceptrum.     

Production of Industrial Enzymes (Amylase,Carboxymethylcellulase and Protease) by Bacteria Isolated from Marine Sedentary Organisms

The abilities of bacteria isolated from eight marine sedentary organisms, six marine sponges (Spirastrella sp., Phyllospongia sp., Ircinia sp., Aaptos sp., Azorica sp. and Axinella sp.), one soft coral (Lobophytum sp.) and one alga (Sargassum sp.) to produce industrial enzymes (amylase, carboxymethylcellulase and protease) were examined. The mean total viable counts of the bacterial isolates ranged from 8.7 × 10⁴ to 8.4 × 10⁵ cfu/g wet weight of the organism. All eight organisms harboured amylase (0.05–0.5 IU/ml), carboxymethylcellulase (0.05–0.5 IU/ml) and protease (0.1–0.5 IU/ml) producing bacteria. Of 56 bacterial strains tested, as many as 60 to 83% of the strains produced at least one of the three enzymes, and 47% of strains were able to produce all three enzymes. High activities (> 0.5 IU/ml) of the three enzymes were recorded in bacterial strains belonging to the genera Alcaligenes and Bacillus. From the results of this study, it appears that bacteria associated with marine sedentary organisms are the novel source of industrial enzymes for possible commercial applications and may play an important role in enzyme‐catalysed organic matter cycling in marine environments.     

The Arabidopsis thaliana Immunophilin ROF1 Directly Interacts with PI(3)P and PI(3,5)P2 and Affects Germination under Osmotic Stress

A direct interaction of the Arabidopsis thaliana immunophilin ROF1 with phosphatidylinositol-3-phosphate and phosphatidylinositol-3,5-bisphosphate was identified using a phosphatidylinositol-phosphate affinity chromatography of cell suspension extracts, combined with a mass spectrometry (nano LC ESI-MS/MS) analysis. The first FK506 binding domain was shown sufficient to bind to both phosphatidylinositol-phosphate stereoisomers. GFP-tagged ROF1 under the control of a 35S promoter was localised in the cytoplasm and the cell periphery of Nicotiana tabacum leaf explants. Immunofluorescence microscopy of Arabidopsis thaliana root tips verified its cytoplasmic localization and membrane association and showed ROF1 localization in the elongation zone which was expanded to the meristematic zone in plants grown on high salt media. Endogenous ROF1 was shown to accumulate in response to high salt treatment in Arabidopsis thaliana young leaves as well as in seedlings germinated on high salt media (0.15 and 0.2 M NaCl) at both an mRNA and protein level. Plants over-expressing ROF1, (WSROF1OE), exhibited enhanced germination under salinity stress which was significantly reduced in the rof1⁻ knock out mutants and abolished in the double mutants of ROF1 and of its interacting homologue ROF2 (WSrof1⁻/2⁻). Our results show that ROF1 plays an important role in the osmotic/salt stress responses of germinating Arabidopsis thaliana seedlings and suggest its involvement in salinity stress responses through a phosphatidylinositol-phosphate related protein quality control pathway.     

Macrophage-mediated cytolysis of erythrocytes in the guinea pig. II. Role of oxidative burst products in macrophage cytotoxicity activated by lectins and phorbol ester derivatives

Guinea pig peritoneal macrophages (GPPM) exhibited enhanced production of O₂^? and H₂O₂, and cytolytic activity toward erythrocytes, in response to reagents such as 12-O-tetradecanoyl-phorbol-13-acetate (TPA), its methylated derivative 4-O-MeTPA, Con A, wheat germ agglutinin (WGA), and opsonized zymosan. In order to examine the possible role of oxidative burst products such as O₂^? and H₂O₂ in the cytolytic process, we used reagents and enzymes which influence the balance of O₂^? and H₂O₂ outside and inside the GPPM cells. Macrophage-mediated cytolysis (MMC) of erythrocytes in the presence of the activators and modulators was assessed by ⁵¹Cr release assay. MMC activated by TPA and 4-O-MeTPA was inhibited by scavengers of H₂O₂ such as catalase and α-tocopherol, and was augmented by the catalase inhibitor 3-amino-1,2,4-triazole, and by horseradish peroxidase. TPA- and 4-O-MeTPA-activated MMC was only partially inhibited by the O₂^? scavenger cytochrome c and the enzyme superoxide dismutase and unaffected by cytochalasin D (an inhibitor of phagocytosis). MMC activated by the lectins Con A and WGA was unaffected by the scavengers and enzymes used, but markedly inhibited by cytochalasin D. Activation of MMC by TPA, WGA, and phagocytosis of opsonized zymosan, as well as O₂^? and H₂O₂ generation triggered by these reagents, were markedly inhibited by chlorpromazine. The results indicate that GPPM-mediated cytolysis activated by lectins, phorbol ester derivatives, and phagocytosis of opsonized zymosan, is dependent on the generation of oxidative burst products, mainly H₂O₂. TPA- or 4-O-MeTPA-activated MMC is mainly an extracellular event, while lectin-activated MMC may take place within the macrophages.     

Taxonomic notes on fruit flies of tribes Schistopterini and Eutretini (Diptera: Tephritidae: Tephritinae) in India,with description of a new species of Rhabdochaeta de Meijere

Rhabdochaeta nigroapicalis David, Hancock and Sachin, sp. n., is described from Assam, NE India. A key to all species of Schistopterini and Eutretini recorded from India is provided and taxonomic notes on previously described species are included. DNA barcode sequences of Calloptera asteria (Schiner), Rhabdochaeta pulchella de Meijere, Rhabdochaeta nigroapicalis David, Hancock and Sachin, sp. n. and Rhochmopterum venustum (de Meijere) were obtained and reported. Phylogenetic analysis using 62 mtCOI sequences of Tephritinae revealed Schistopterini to be a monophyletic group and the new species closely related to Rhabdochaeta pulchella, justifying its placement in Rhabdochaeta de Meijere.http://zoobank.org/A97337B3-E2BD-460D-BA62-6D3D4A1A60A2     

Inhibition of Quorum Sensing Mediated Virulence Factors Production in Urinary Pathogen Serratia marcescens PS1 by Marine Sponges

The focal intent of this study was to find out an alternative strategy for the antibiotic usage against bacterial infections. The quorum sensing inhibitory (QSI) activity of marine sponges collected from Palk Bay, India was evaluated against acyl homoserine lactone (AHL) mediated violacein production in Chromobacterium violaceum (ATCC 12472), CV026 and virulence gene expressions in clinical isolate Serratia marcescens PS1. Out of 29 marine sponges tested, the methanol extracts of Aphrocallistes bocagei (TS 8), Haliclona (Gellius) megastoma (TS 25) and Clathria atrasanguinea (TS 27) inhibited the AHL mediated violacein production in C. violaceum (ATCC 12472) and CV026. Further, these sponge extracts inhibited the AHL dependent prodigiosin pigment, virulence enzymes such as protease, hemolysin production and biofilm formation in S. marcescens PS1. However, these sponge extracts were not inhibitory to bacterial growth, which reveals the fact that the QSI activity of these extracts was not related to static or killing effects on bacteria. Based on the obtained results, it is envisaged that the marine sponges could pave the way to prevent quorum sensing (QS) mediated bacterial infections.     

Proteomics and Metabolomics Analyses to Elucidate the Desulfurization Pathway of Chelatococcus sp.

Desulfurization of dibenzothiophene (DBT) and alkylated DBT derivatives present in transport fuel through specific cleavage of carbon-sulfur (C-S) bonds by a newly isolated bacterium Chelatococcus sp. is reported for the first time. Gas chromatography-mass spectrometry (GC-MS) analysis of the products of DBT degradation by Chelatococcus sp. showed the transient formation of 2-hydroxybiphenyl (2-HBP) which was subsequently converted to 2-methoxybiphenyl (2-MBP) by methylation at the hydroxyl group of 2-HBP. The relative ratio of 2-HBP and 2-MBP formed after 96 h of bacterial growth was determined at 4:1 suggesting partial conversion of 2-HBP or rapid degradation of 2-MBP. Nevertheless, the enzyme involved in this conversion process remains to be identified. This production of 2-MBP rather than 2-HBP from DBT desulfurization has a significant metabolic advantage for enhancing the growth and sulfur utilization from DBT by Chelatococcus sp. and it also reduces the environmental pollution by 2-HBP. Furthermore, desulfurization of DBT derivatives such as 4-M-DBT and 4, 6-DM-DBT by Chelatococcus sp. resulted in formation of 2-hydroxy-3-methyl-biphenyl and 2-hydroxy –3, 3^/- dimethyl-biphenyl, respectively as end product. The GC and X-ray fluorescence studies revealed that Chelatococcus sp. after 24 h of treatment at 37°C reduced the total sulfur content of diesel fuel by 12% by per gram resting cells, without compromising the quality of fuel. The LC-MS/MS analysis of tryptic digested intracellular proteins of Chelatococcus sp. when grown in DBT demonstrated the biosynthesis of 4S pathway desulfurizing enzymes viz. monoxygenases (DszC, DszA), desulfinase (DszB), and an NADH-dependent flavin reductase (DszD). Besides, several other intracellular proteins of Chelatococcus sp. having diverse biological functions were also identified by LC-MS/MS analysis. Many of these enzymes are directly involved with desulfurization process whereas the other enzymes/proteins support growth of bacteria at an expense of DBT. These combined results suggest that Chelatococcus sp. prefers sulfur-specific extended 4S pathway for deep-desulphurization which may have an advantage for its intended future application as a promising biodesulfurizing agent.     

Lethal and non-lethal effects of an invasive naticid gastropod on the production of a native clam

Predators often affect prey production not only by lethal predation but also by unintentional inhibition of feeding and growth. The present study examined the lethal and non-lethal effects of the invasive naticid Laguncula pulchella on the survival and growth of the prey clam Ruditapes philippinarum in a sandy tidal flat. Cages accommodating 30 clams (10 individuals × 3 size classes of ≤ 20 mm, 20–30 mm, and > 30 mm shell length) and one L. pulchella (approximately 37 mm shell height) per cage were buried in the tidal flat for 10 weeks. Medium sized clams were consumed by predators much more (80.5%) than small (12.2%) and large clams (7.3%). Clams were consumed by L. pulchella at a frequency of 0–2.5 individuals per predator per week. The growth of clams caged with L. pulchella was lower (23, 27, 33, 41, and 57% for clam of 10, 20, 30, 40, and 50 mm, respectively) than that in control cages (clams without L. pulchella). The clam burial depths did not increase by the presence of predators in a laboratory experiment, indicating that growth suppression was caused by the reduced feeding activity following physical disturbance and/or chemical signals. The results of this study demonstrate that the introduction of L. pulchella reduced the productivity of the commercially important clams not only by lethal predation but also by mere presence.     

Fatty Acid and Phospholipid Syntheses Are Prerequisites for the Cell Cycle of Symbiodinium and Their Endosymbiosis within Sea Anemones

Lipids are a source of metabolic energy, as well as essential components of cellular membranes. Although they have been shown to be key players in the regulation of cell proliferation in various eukaryotes, including microalgae, their role in the cell cycle of cnidarian-dinoflagellate (genus Symbiodinium) endosymbioses remains to be elucidated. The present study examined the effects of a lipid synthesis inhibitor, cerulenin, on the cell cycle of both cultured Symbiodinium (clade B) and those engaged in an endosymbiotic association with the sea anemone Aiptasia pulchella. In the former, cerulenin exposure was found to inhibit free fatty acid (FFA) synthesis, as it does in other organisms. Additionally, while it also significantly inhibited the synthesis of phosphatidylethanolamine (PE), it did not affect the production of sterol ester (SE) or phosphatidylcholine (PC). Interestingly, cerulenin also significantly retarded cell division by arresting the cell cycles at the G₀/G₁ phase. Cerulenin-treated Symbiodinium were found to be taken up by anemone hosts at a significantly depressed quantity in comparison with control Symbiodinium. Furthermore, the uptake of cerulenin-treated Symbiodinium in host tentacles occurred much more slowly than in untreated controls. These results indicate that FFA and PE may play critical roles in the recognition, proliferation, and ultimately the success of endosymbiosis with anemones.