Comparison of cellulose solubilisation rates in rumen and landfill leachate inoculated reactors

The aim of this study was to conduct a number of controlled digestions to obtain easily comparable cellulose solubilisation rates and to compare these rates to those found in the literature to see which operational differences were significant in affecting cellulose degradation during anaerobic digestion. The results suggested that differences in volumetric cellulose solubilisation rates were not indicative of the true performance of cellulose digestion systems. When cellulose solubilisation rates were normalised by the mass of cellulose in the reactor at each time step, the comparison of the rates became more meaningful. Cellulose solubilisation was surface area limited. Therefore, changes in the loading rate of cellulose to the reactor altered the volumetric solubilisation rate without changing the mass normalised rate. Comparison of mass normalised solubilisation rates from this study and the literature demonstrated that differences in reactor configuration and operational conditions did not significantly impact on the solubilisation rate whereas the difference in composition of the microbial communities showed a marked effect. This work highlights the importance of using appropriately normalised data when making comparisons between systems with differing operational conditions. (c) 2005 Elsevier Ltd. All rights reserved.

Development of an Efficient Genome Editing Method by CRISPR/Cas9 in a Fish Cell Line

This project was carried out during CD’s BBSRC Eastbio PIPS placement at Marine Scotland. The authors are grateful to Dr Milena Monte (University of Aberdeen) for help with the FACS analysis. The authors wish to thank Dr Filippo Del Bene (Neuronal Circuit Development, Institut Curie) and Dr Wenbiao Chen (School of Medicine, Vanderbilt University) for the Addgene plasmids, #61051 and #47929, respectively, and Prof. Nancy C. Reich Marshall (Department of Molecular Genetics and Microbiology, Stony Brooks University) for the plasmid pmEGFP-N1.

Development of an Efficient Genome Editing Method by CRISPR/Cas9 in a Fish Cell Line

This project was carried out during CD’s BBSRC Eastbio PIPS placement at Marine Scotland. The authors are grateful to Dr Milena Monte (University of Aberdeen) for help with the FACS analysis. The authors wish to thank Dr Filippo Del Bene (Neuronal Circuit Development, Institut Curie) and Dr Wenbiao Chen (School of Medicine, Vanderbilt University) for the Addgene plasmids, #61051 and #47929, respectively, and Prof. Nancy C. Reich Marshall (Department of Molecular Genetics and Microbiology, Stony Brooks University) for the plasmid pmEGFP-N1.

Cytotoxicity Evaluation of Si-Diatom Frustules in a Mouse Model

Silica additives in bone substitute materials are topical, clinically interesting and have significant support in the Orthopaedic field. Biosilica, e.g isolated from diatoms, has many advantages over its synthetic counterparts, e.g. it is amorphous, thus will be absorbed by the body, however, issues such as purity, presence of endotoxins and cytotoxicity need to be addressed before it can be further exploited. Biosilica isolated from Cyclotella Meneghiniana was then tested in a mouse model, to test the immunological response, organ toxicity (kidney, spleen, liver) and route of metabolism/excretion of silica. Five-week-old Balb-c mice were injected subcutaneously with a single high dose (50mg/ml) of Si-frustules, Si-frustules + organic linker and vehicle only control. Animals were sacrificed at 1d and 28d. The animal studies were conducted under an ethically approved protocol at Queen’s University, Belfast. The animals showed no adverse stress during the experiment and remained healthy until sacrifice. Blood results using ICP-OES analysis suggest the frustules were metabolized between comparator groups at different rates, and clearly showed elevated levels of silicon in groups injected with frustules relative to control. The histology of organs showed no variation in morphology of mice injected frustules relative compared to the control group.
Acknowledgements: The authors would like to thank Marie Curie International Outgoing Fellowships from the EU and Beaufort Marine Biodiscovery Award as part of the Marine Biotechnology Ireland Programme for providing financial support to this project.

Molecular biotechnology of marine algae in China

Molecular biotechnology of marine algae is referred to as the biotechnology on the identification, modification, production and utilization of marine algal molecules. It involves not only the manipulation of macromolecules such as DNA, RNA and proteins, but also deals with low molecular weight compounds such as secondary metabolites. In the last decade, molecular systematic researches to investigate the relationship and to examine the evolutionary divergence among Chinese marine algae have been carried out by Chinese scientists. For example, RAPD has been widely used in several laboratories to elucidate genetic variations of the reds, such as Porphyra, Gracilaria, Grateloupia and the greens such as Ulva and Enteromorpha. Some important data have been obtained. The study on molecular genetic markers for strain improvement is now in progress. In 1990s, genetic engineering of economic seaweeds such as Laminaria, Undaria, Porphyra, Gracilaria and Grateloupia has been studied in China. For Laminaria japonica, the successfully cultivated kelp in China, a model transformation system has been set up based on the application of plant genetic techniques and knowledge of the algal life history. Progress has been made recently in incorporating a vaccine gene into kelp genome. Evidence has been provided showing the expression of gene products as detectable vaccines. In the present paper, the progress of molecular biotechnological studies of marine algae in China, especially researches on elucidating and manipulating nucleic acids of marine algae, are reviewed.

Extracellular beta-Glucosidase production by marine Aspergillus sydowii BTMFS 55

The beta-glucosidase enzyme purified from the marine fungus, Aspergillus sydowii BTMFS 55 showed a good yield of enzyme production under solid state fermentation. The statistical optimization of the media components revealed that moisture content, concentration of peptone and inoculum are the major parameters which supported the maximal enzyme production. The purified enzyme showed low pH activity and stability, glucose tolerance and activation by ethanol. It could produce ethanol from wheat bran and rice straw by simultaneous saccharification and fermentation with yeast.The glucosidase purified from Aspergillus sydowii BTMFS 55 shows great potential for several biotechnological applications such as the production of bio-ethanol from agricultural biomass and improvement in the aromatic character of wines and fruit juices through the hydrolysis of flavour glucosidic precursors. There is immense scope for the application of this marine fungus in the biofuel production besides in other industries provided further studies are pursued in exploiting this enzyme and the organism particularly scale up studies with respect to application. There is also ample scope for cloning of the gene encoding beta-glucosidase in domesticated hosts such as Pichia pastoris or S. cerevisiae that can produce ethanol directly from cellulosic biomass.

Sustainable production of biologically active molecules of marine based origin

The marine environment offers both economic and scientific potential which are relatively untapped from a biotechnological point of view. These environments whilst harsh are ironically fragile and dependent on a harmonious life form balance. Exploitation of natural resources by exhaustive wild harvesting has obvious negative environmental consequences. From a European industry perspective marine organisms are a largely underutilised resource. This is not due to lack of interest but due to a lack of choice the industry faces for cost competitive, sustainable and environmentally conscientious product alternatives. Knowledge of the biotechnological potential of marine organisms together with the development of sustainable systems for their cultivation, processing and utilisation are essential. In 2010, the European Commission recognised this need and funded a collaborative RTD/SME project under the Framework 7-Knowledge Based Bio-Economy (KBBE) Theme 2 Programme 'Sustainable culture of marine microorganisms, algae and/or invertebrates for high value added products'. The scope of that project entitled 'Sustainable Production of Biologically Active Molecules of Marine Based Origin' (BAMMBO) is outlined. Although the Union is a global leader in many technologies, it faces increasing competition from traditional rivals and emerging economies alike and must therefore improve its innovation performance. For this reason innovation is placed at the heart of a European Horizon 2020 Strategy wherein the challenge is to connect economic performance to eco performance. This article provides a synopsis of the research activities of the BAMMBO project as they fit within the wider scope of sustainable environmentally conscientious marine resource exploitation for high-value biomolecules. © 2013 Elsevier B.V.

A suite of novel promoters and terminators for plant biotechnology

The gene regulation signals from subterranean clover stunt virus (SCSV) were investigated for their expression in dicot plants. The SCSV genome has at least eight circular DNA molecules. Each circular DNA component contains a promoter element, a single open reading frame and a terminator. The promoters from seven of the segments were examined for their strength and tissue specificity in transgenic tobacco (Nicotiana tabacum L.), potato (Solanum tuberosum L.) and cotton (Gossypium hirsutum L.) using a GUS reporter gene assay system. While the promoters of many of the segments were poorly expressed, promoters derived from segments 4 and 7 were shown to direct high levels of expression in various plant tissues and organs. The segment 1 promoter directs predominantly callus-specific expression and, when used to control a selectable marker gene, facilitated the transformation of all three species (tobacco, potato and cotton). From the results, a suite of plant expression vectors (pPLEX) derived from the SCSV genome were constructed and used here to produce herbicide- and insect-resistant cotton, demonstrating their utility in the expression of foreign genes in dicot crop species and their potential for use in agricultural biotechnology.

Quantitative assessment of total and Gram-positive aerobic bacteria in fresh and ambient-temperature-stored sub-tropical marine fish

The current study was undertaken to enumerate Gram-positive bacteria in fresh sub-tropical marine fish and determine the effect of ambient storage (25°C) on the Gram-positive bacterial count. Total and Gram-positive bacteria were enumerated in the muscles, gills and gut of fresh and stored Pseudocaranx dentex, Pagrus auratus and Mugil cephalus on tryptone soya agar (TSA) and TSA with 0.25% phenylethyl alcohol (PEA), respectively. Initial studies indicated that PEA significantly reduced total aerobic bacterial count (TABC) whereas control Gram-positive bacteria were not affected by 0.25% PEA. TABC significantly increased in all fish body parts, whereas Gram-positive aerobic bacterial count (GABC) significantly increased only in the muscles and gills during ambient storage for 15 h. The TABC of the fish species increased from 4.00, 6.13 and 4.58 log cfu g-1, respectively in the muscles, gills, and gut to 6.31, 7.31 and 7.23 log cfu g-1 by the end of storage. GABC increased from 2.00, 3.52 and 2.20 log cfu g-1 to 4.70, 5.85 and 3.36 log cfu g-1. Within each species, TABC were significantly higher in the gills compared to that of muscles and gut; however, no significant differences were found in GABC between muscles and gills. This study demonstrated the potential importance of Gram-positive bacteria in sub-tropical marine fish and their spoilage.

Seagrass-Watch: Engaging Torres Strait Islanders in marine habitat monitoring

Involvement in scientifically structured habitat monitoring is a relatively new concept to the peoples of Torres Strait. The approach we used was to focus on awareness, and to build the capacity of groups to participate using Seagrass-Watch as the vehicle to provide education and training in monitoring marine ecosystems. The project successfully delivered quality scientifically rigorous baseline information on the seasonality of seagrasses in the Torres Strait-a first for this region. Eight seagrass species were identified across the monitoring sites. Seagrass cover varied within and between years. Preliminary evidence indicated that drivers for seagrass variability were climate related. Generally, seagrass abundance increased during the north-west monsoon (Kuki), possibly a consequence of elevated nutrients, lower tidal exposure times, less wind, and higher air temperatures. Low seagrass abundance coincided with the presence of greater winds and longer periods of exposure at low tides during the south-east trade wind season (Sager). No seasonal patterns were apparent when frequency of disturbance from high sedimentation and human impacts was high. Seagrass-Watch has been incorporated in to the Thursday Island High School's Marine Studies Unit ensuring continuity of monitoring. The students, teachers, and other interested individuals involved in Seagrass-Watch have mastered the necessary scientific procedures to monitor seagrass meadows, and developed skills in coordinating a monitoring program and skills in mentoring younger students. This has increased the participants' self-esteem and confidence, and given them an insight into how they may participate in the future management of their sea country.

Chronicle of Marine Diatom Culturing Techniques

Diatoms are regarded as useful neutral lipid sources, as liquid fuel precursors, as foods for marine culture of zooplankters, larval and post-larval shrimp, copepods, juvenile oysters and as micromachines in nanotechnology. Combining microscopic observation with in situ culturing has been useful in areas of taxonomy, ecology, biomonitoring, biotechnology, etc. This communication reviews various culturing techniques of marine diatoms with the relative merits.

Agricultural Best Management Practices and Treatment Wetlands in the Gabilan Watershed: Project Asessment and Evaluation Plan

Several local groups have come together for this project to addresses water quality concerns in the Gabilan Watershed – also known as the Reclamation Ditch Watershed (Fig. 1.1). These are Moss Landing Marine Laboratories (MLML), the Resource Conservation District of Monterey County (RCDMC), Central Coast Watershed Studies (CCoWS), Return of the Natives (RON), Community Alliance with Family Farmers (CAFF), and Coastal Conservation and Research (CC&R). The primary goal is to reduce non-point source pollution – particularly suspended sediment, nutrients, and pesticides – and thereby improve near-shore coastal waters of Moss Landing Harbor and the Monterey Bay. (Document contains 33 pages)