Studies on habitat selection by amphipods using artificial substrates within an estuarine environment

Field studies on habitat selection and community development of four gammarid amphipod species were conducted in a brackish lagoon at Nanakita River Estuary, Japan. Development of amphipod community was studied by focussing mainly on provision of shelter for these invertebrates. Seven artificial substrates (including wire net cage, circular concrete block, dried oyster shell, brush, sponge, blue and green tapes) were used in the field as substitutes of natural substrates (macroalgae). This paper demonstrates that amphipods will mostly select and inhabit substrates with available space that provide them with adequate protection. Brush substrate was ranked the best among the selected substrates, because its filamentous nature like natural substrates (macroalgae) provided suitable habitat for both tube-dwelling and free-living amphipods and accommodated maximum densities of amphipods in all seasons at all stations. Tube-dwelling amphipods (domicolous) constructed their tubes in and on algal substrates and their densities tended to be correlated with algal biomass, whilst free-living species (nestlers) prowled on surfaces, clang to filamentous structures and hid themselves in crevices, holes and boulders. Brush substrates at different stations suggested the importance of site difference.     

Experimental and physiological ecology of ammonia fungi: studies using natural substrates and artificial media

Experiments using natural substrates and artificial media were categorized into three groups. Using experiments in these three categories, we investigated dispersion, invasion sequence, and decomposing ability of ammonia fungi in the field on natural substrates. Spore germination and vegetative and reproductive growth of ammonia fungi derived from monocultures and five-species cultures were assessed on natural substrates and on artificial media. These features assist understanding of the colonization and successive occurrence of ammonia fungi in the field based on estimation of interactions among the fungi and the physiological characteristics of each ammonia fungus. The sequential colonization and fruiting of ammonia fungi in the field can be explained primarily by the preference or tolerance of a species to high concentrations of ammonium-N under alkaline to neutral conditions. Succession of ammonia fungi results from their sequential colonization associated with the time needed for each fungus to produce reproductive structures. Colonization likely initiates from mycelia and/or spores preinhabiting the soil and from newly invading airborne propagules. Duration of occurrence of ammonia fungi in the field is shortened by the interactions among them. Saprobic ammonia fungi are the principal agents of litter decomposition under alkaline to neutral conditions. Parts of this review were presented as the presidential address of the Mycological Society of Japan (MSJ) at the Mycological Society of America (MSA) and MSJ Joint Meeting 2005 held in Hawaii in August 2005     

A multi-lake comparison of epilithic diatom communities on natural and artificial substrates

In a multi-season, multi-lake study of epilithic diatom communities, glass slide artificial substrate samplers provided poor representation of communities on natural substrates. Percent similarities between the two communities averaged only 37 over the course of the study. Overall, natural substrates exhibited greater species richness than artificial samplers, particularly during the summer, although this difference was slight. Patterns of selection and inhibition by artificial substrates of individual diatom species varied by both season and lake. Members of the genus Cymbella, particularly C. microcephala, appeared to be the most consistently inhibited, while Achnanthes minutissima was often selected for. In spite of the great differences between the two substrate types, replicability of artificial substrates was very high, and could prove to be a more important quality in monitoring applications than accurate representation of the natural community.     

Studies on sphingomyelinase and beta-glucosidase activities in Niemann-Pick disease variants. Phosphodiesterase activities measured with natural and artificial substrates

Cultured fibroblasts were studied from 12 cases of Niemann-Pick disease group C. In 11, sphingomyelinase and glucocerebrosidase (and beta-glucosidase) activities were reduced to around 50% of those of controls. On isoelectric focusing, all 12 strains lacked sphingomyelinase activity in the major cathodic region (pI 8.0). The defect was also demonstrated with the artificial phosphodiester substrates bis(4-methylumbelliferyl) phosphate and 4-methylumbelliferyl pyrophosphate diester. In control fibroblasts and those heterozygous for types A or B or group C Niemann-Pick disease, the major sphingomyelinase peak electrofocused at pI 8.0. No direct interaction could be demonstrated by mixing experiments between group C Niemann-Pick extracts and those of type A disease or Gaucher disease. Profiles for beta-glucosidase activity appeared normal in Niemann-Pick group C fibroblasts. No reduction of sphingomyelinase or glucocerebrosidase activities was found in Niemann-Pick group C liver, nor any attenuation of cathodic sphingomyelinase activity in the affected tissue. Results suggest that sphingomyelinase expression differs in fibroblasts and liver. Enzyme defects associated with Niemann-Pick disease group C were only observed in cultured cells.     

Specificity of natural and artificial substrates for human Cdc25A

Cdc25A is a dual-specific protein phosphatase involved in the regulation of the kinase activity of Cdk-cyclin complexes in the eukaryotic cell cycle. To understand the mechanism of this important regulator, we have generated highly purified biochemical reagents to determine the kinetic constants for human Cdc25A with respect to a set of peptidic, artificial, and natural substrates. Cdc25A and its catalytic domain (dN25A) demonstrate very similar kinetics toward the artificial substrates p-nitrophenyl phosphate (k(cat)/K(m) = 15-25 M(-1) s(-1)) and 3-O-methylfluorescein phosphate (k(cat)/K(m) = 1.1-1.3 x 10(4) M(-1) s(-1)). Phospho-peptide substrates exhibit extremely low second-order rate constants and a flat specificity profile toward Cdc25A and dN25A (k(cat)/K(m) = 1 to 10 M(-1) s(-1)). In contrast to peptidic substrates, Cdc25A and dN25A are highly active phosphatases toward the natural substrate, T14- and Y15-bis-phosphorylated Cdk2/CycA complex (Cdk2-pTpY/CycA) with k(cat)/K(m) values of 1.0-1.1 x 10(6) M(-1) s(-1). In the context of the Cdk2-pTpY/CycA complex, phospho-threonine is preferred over phospho-tyrosine by more than 10-fold. The highly homologous catalytic domain of Cdc25c is essentially inactive toward Cdk2-pTpY/CycA. Taken together these data indicate that a significant degree of the specificity of Cdc25 toward its Cdk substrate resides within the catalytic domain itself and yet is in a region(s) that is outside the phosphate binding site of the enzyme.     

Chick embryo liver beta-glucuronidase. Comparison of activity on natural and artificial substrates.

The beta-glucuronidase in homogenates of 12-day chick embryo livers catalyzed the release of glucuronic acid from 4-methylumbelliferyl-beta-D-glucuronide and from the nonreducing terminals of the hexasaccharides of chondroitin-6-SO4 and chondroitin-4-SO4 at rates of 143, 114, and 108 nmol of glucuronic acid/h/mg of protein, respectively, when assayed at pH 3.5 in 0.05 M sodium acetate buffer. During a 60-fold purification of the enzyme, the ratios of the activities on these substrates did not change. When 4-methylumbelliferyl-beta-D-glucuronide was used as substrate the enzyme was active at pH values from 3.0 to 5.5, with maximal activity between pH values 4.0 and 4.5. Concentrations of NaCl from 0.15 to 0.3 M inhibited the activity at low pH values but activated the enzyme between pH 4.0 and 5.5. The enzyme was active on the chondroitin-6-SO4 hexasaccharide from pH 3.0 to 5.5, with a broad optimum between 3.0 and 4.5. NaCl inhibited the activity on the oligosaccharide substrate at all pH values. Eadie-Scatchard plots of rates of 4-methylumbelliferyl-beta-D-glucuronide hydrolysis at substrate concentrations ranging from 2 to 1000 microM showed multiple kinetic forms of the enzyme, a form with a Km of approximately 11 microM, and a second form with a Km of approximately 225 microM. The pH optimum of the low Km form was 3.5 to 4.0; that of the high Km form was pH 4.5. NaCl inhibited the activity of the low Km form, but activated the high Km form of the enzyme. Chondroitin SO4 oligosaccharides competed with 4-methylumbelliferyl-beta-D-glucuronide for the low Km form of the enzyme but had little effect on the hydrolysis of 4-methylumbelliferyl-beta-D-glucuronide by the high Km form of the enzyme. The activities of the beta-glucuronidase on tetra-, hexa-, octa-, and decasaccharides of chondroitin-6-SO4 and chondroitin-4-SO4, measured using a new assay procedure which can detect the formation of 1 nmol of product, were similar, although rates were somewhat lower for the higher oligosaccharides. With the exception of the chondroitin-4-SO4 tetrasaccharide, all of the oligosaccharide substrates saturated the enzyme at concentrations of 20 to 30 microM, indicating Km values of less than 10 to 15 microM for the oligosaccharides. Highly purified beta-glcuronidases from human placenta and from rat preputial gland also showed multiple kinetic forms when assayed using 4-methylumbelliferyl-beta-D-glucuronide as substrate.     

Observations on diatoms inhabiting natural and artificial substrates in Kongsfjorden,Svalbard, the Arctic

Polar Biology - The most dramatic effects of global climate change are predicted for the Arctic, and there is a raising concern about the lack of baseline information on microalgal biodiversity....     

Studies in the life cycle of Allomyces javanicus Kniep

Summary The life cycle of Allomyces javanicus was studied with the hanging drop method under laboratory conditions. The isolate has a life cyrle similar to that already described in A. javanicus and A. arbuscula. Planonts from resistant sporangia do not germinate directly to produce gametophytic plants, but on the contrary, the latter germinate to produce plants like their immediate parents. Female and male gametes growing in the gametangia of relatively young sexual mycelia develop into asexual mycelia after conjugation, but some of the female gametes after germination develop into asexual mycelia without conjugation.     

Vertical profiles of current velocity and dissolved oxygen saturation in biofilms on artificial and natural substrates

We determined vertical changes in current velocity and dissolved oxygen concentration in biofilms on artificial and natural substrates using microelectrodes. We used biofilms developed on glass slides dipped in an artificial stream for 3 months, artificial clay tiles dipped in an outdoor artificial stream for 3 months, and natural pebbles. In the biofilm on a glass slide, current velocity significantly decreased from the surface of the biofilm and became 0 cm s⁻¹ at the surface of the glass slide. Vertical profile of current velocity versus depth indicated a presence of a viscous sublayer of 0.2-mm thickness above the surface of glass slide. Dissolved oxygen (DO) concentration increased within the biofilm and attained the maximum (123%) at the surface of the glass slide, indicating active photosynthesis by sessile diatoms at the layer corresponding to the observed viscous sublayer. In the biofilm on an artificial tile, DO increased to 163% saturation at 24.849-mm depth, followed by rapid decrease (6%) at the surface of the tile. A similar result of remarkable decrease in DO saturation was also found in the biofilm on a natural pebble. These results suggest that smoothness of the substrate surface is related to the vertical profile of DO saturation. The thickness of the viscous sublayer and oxygen-depleted area (up to several hundred micrometers, μm) was sufficient for the presence of bacteria, protists, and other metazoan animals, suggesting high activity and diversity of those heterotrophs in the bottom part of biofilms.     

Studies on conidial germination and initial growth of the grapevine powdery mildew Uncinula necator on artificial substrates

Summary The requirements for conidial germination and formation of infection structures of the grapevine parasites Uncinula necator were studied on different substrates and supports. Direct contact with water or agar surfaces frequently caused bursting of conidia or inhibited formation of infection structures. Normal germination and initial growth depended on the presence of a dry surface covering a liquid substrate which could be reached by the penetration pegs. The water necessary for germination must be provided by a high air humidity whose optimal value (99.8%) was determined using closed slide microchambers in which air humidity was controlled by salt solutions. Even 99.6% humidity led to significant growth reduction, while at higher values deleterious condensation droplets formed around the spores. If liquid substrates are covered with thin (0.5 m), waterproof polyacrylic membranes, germination appears normal. Formation of appressoria, successful penetration of the membrane by infection pegs and the development of functional haustoria indicate a lack of host specifity during the initial growth stage. although on water and on glucose media growth stopped after 1–2 weeks, this method seems to indicate a way to axenically culture this obligate parasite. The lack of host specificity at the level of host surface or tissue anatomy was corroborated by a successful culture of powdery mildew on grapevine callus.Dedicated to Professor Dr. Dr. hc. K. Esser on the occasion of his 65th anniversary     

Comparison of temperature and moisture requirements for sporulation of Aspergillus flavus sclerotia on natural and artificial substrates

A key step in the infection cycle by Aspergillus flavus in maize is sporulation of sclerotia present in soil or in crop debris. However, little information is available on this critical and important phase. This study included experiments on artificial (Czapek Dox Agar (CZ)) and natural (maize stalks) substrates under different conditions of temperature (T; from 5 to 45 °C) and water activity (a(w); from 0.50 to 0.99) levels to quantify sporulation from sclerotia. The mean numbers of spores were higher on defined nutritional medium in vitro on CZ agar than on maize stalks (4.5×10(6) spores/sclerotium versus 4.2×10(4) spores/sclerotium) with production initiated after 6 and 24h, respectively. Surprisingly, the optimal temperature was found at 30-35 °C for CZ agar (9.23×10(6) spores/sclerotium) and to be 20-25 °C for maize stalks (6.26×10(4) spores/sclerotium). Water stress imposition only reduced sporulation at ≤0.90 a(w.) With more available water no significant differences were found between 0.90 and 0.99 a(w). This type of data is critical in the development of a mechanistic model to predict the infection cycle of A. flavus in maize in relation to meteorological conditions.     

Gas exchange and nutrition patterns during the life cycle of an artificial wheat crop

The growth, mineral and shoot and root CO₂ exchange of wheat plants ( Triticum aestivum L. cv. Courtot) cultivated in growth chambers have been studied during the complete life cycle. The life cycle could be divided into 4 periods according to the patterns of CO₂ exchange: exponential increase, linear increase, stabilization and decline of photosynthesis. These patterns are analysed in relation to light interception, tillering, competition between plants and ageing of the leaves, all of which constitute successive limiting factors. Root metabolism seemed to be subordinated to the demand of the shoot for minerals. Ion uptake from the nutrient solution was particularly pronounced in young plants, which were higher in minerals and nitrogen than older ones. The ratios of K and P uptake varied with plant age, and the charge balance in ion exchange was equlibrated by H uptake. Rhythmic patterns appeared in all exchanges and varied with plant age. The highest amplitude of rhythm was found in root respiration. After taking the losses due to respiration and photorespiration into account, the maximum rate of photosynthesis approached the theoretical value calculated from the light energy absorbed.     

Liquefied natural gas for the UK: a life cycle assessment

Purpose

Liquefied natural gas (LNG) is expected to become an important component of the UK’s energy supply because the national hydrocarbon reserves on the continental shelf have started diminishing. However, use of any carbon-based fuel runs counter to mitigation of greenhouse gas emissions (GHGs). Hence, a broad environmental assessment to analyse the import of LNG to the UK is required.

Methods

A cradle to gate life cycle assessment has been carried out of a specific but representative case: LNG imported to the UK from Qatar. The analysis covers the supply chain, from gas extraction through to distribution to the end-user, assuming state-of-the-art facilities and ships. A sensitivity analysis was also conducted on key parameters including the energy requirements of the liquefaction and vaporisation processes, fuel for propulsion, shipping distance, tanker volume and composition of raw gas.

Results and discussion

All environmental indicators of the CML methodology were analysed. The processes of liquefaction, LNG transport and evaporation determine more than 50% of the cradle to gate global warming potential (GWP). When 1% of the total gas delivered is vented as methane emissions leakage throughout the supply chain, the GWP increases by 15% compared to the GWP of the base scenario. The variation of the GWP increases to 78% compared to the base scenario when 5% of the delivered gas is considered to be lost as vented emissions. For all the scenarios analysed, more than 75% of the total acidification potential (AP) is due to the sweetening of the natural gas before liquefaction. Direct emissions from transport always determine between 25 and 49% of the total eutrophication potential (EP) whereas the operation and maintenance of the sending ports strongly influences the fresh water aquatic ecotoxicity potential (FAETP).

Conclusions

The study highlights long-distance transport of LNG and natural gas processing, including sweetening, liquefaction and vaporisation, as the key operations that strongly affect the life cycle impacts. Those cannot be considered negligible when the environmental burdens of the LNG supply chain are considered. Furthermore, the effect of possible fugitive methane emissions along the supply chain are critical for the impact of operations such as extraction, liquefaction, storage before transport, transport itself and evaporation.

     

Studies on the catalytic domains of multiple JmjC oxygenases using peptide substrates

The JmjC-domain-containing 2-oxoglutarate-dependent oxygenases catalyze protein hydroxylation and N^?-methyllysine demethylation via hydroxylation. A subgroup of this family, the JmjC lysine demethylases (JmjC KDMs) are involved in histone modifications at multiple sites. There are conflicting reports as to the substrate selectivity of some JmjC oxygenases with respect to KDM activities. In this study, a panel of modified histone H3 peptides was tested for demethylation against 15 human JmjC-domain-containing proteins. The results largely confirmed known N^?-methyllysine substrates. However, the purified KDM4 catalytic domains showed greater substrate promiscuity than previously reported (i.e., KDM4A was observed to catalyze demethylation at H3K27 as well as H3K9/K36). Crystallographic analyses revealed that the N^?-methyllysine of an H3K27me3 peptide binds similarly to N^?-methyllysines of H3K9me3/H3K36me3 with KDM4A. A subgroup of JmjC proteins known to catalyze hydroxylation did not display demethylation activity. Overall, the results reveal that the catalytic domains of the KDM4 enzymes may be less selective than previously identified. They also draw a distinction between the N^?-methyllysine demethylation and hydroxylation activities within the JmjC subfamily. These results will be of use to those working on functional studies of the JmjC enzymes.     

Studies on the life cycle and reproduction of the parasitic amphipodHyperia galba in the North Sea

The structure of aHyperia galba population, and its seasonal fluctuations were studied in the waters of the German Bight around the island of Helgoland over a period of two years (1984 and 1985). A distinct seasonal periodicity in the distribution pattern of this amphipod was recorded. During summer, when its hosts—the scyphomedusaeAurelia aurita, Chrysaora hysoscella, Rhizostoma pulmo, Cyanea capillata andCyanea lamarckii—occur in large numbers, supplying shelter and food, a population explosion ofH. galba can be observed. It is caused primarily by the relatively high fecundity ofH. galba which greatly exceeds that of other amphipods: a maximum of 456 eggs was observed. The postembryonic development is completed in the medusae infested; only then are the young able to swim and search for a new host. The smallest fréely-swimming hyperians obtained from plankton samples were 2.6 mm in body size. The size classes observed as well as moult increment and moulting frequencies in relation to different temperatures suggest that two generations are developed per year: a rapidly growing generation in summer and a slower growing generation in winter that shifts to a benthic mode of life and hibernation. For short periods, adult hyperians may become attached to zooplankters other than scyphomedusae. However, when releasing the progeny, they are dependent on the presence of these coelenterates. Apparently, a host specificity does not exist. During daytime, the hyperians seem to avoid a host change; only 0.2% of all the individuals sampled in the plankton during the day were not associated with medusae. The heavy infestation of medusae by this crustacean leads to a weakening and a progressive breakdown of these important predators on fish larvae.H. galba occupies a specific position in the marine food web which is discussed in detail.