Molecular cloning of a novel putative potassium channel-blocking neurotoxin from the venom of the North African scorpion, Androctonus amoreuxi

Scorpion venoms are a particularly rich source of neurotoxic proteins/peptides that interact in a highly specific fashion with discrete subtypes of ion channels in excitable and non-excitable cells. Here we have employed a recently developed technique to effect molecular cloning and structural characterization of a novel putative potassium channel-blocking toxin from the same sample of venom from the North African scorpion, Androctonus amoreuxi. The deduced precursor open-reading frame is composed of 59 amino acid residues that consists of a signal peptide of approximately 22 amino acid residues followed by a mature toxin of 37 amino acid residues. The mature toxin contains two functionally important residues (Lys27 and Tyr36), constituting a functional dyad motif that may be critical for potassium channel-blocking activity that can be affirmed from structural homologs as occurring in the venoms from other species of Androctonus scorpions. Parallel proteomic/transcriptomic studies can thus be performed on the same scorpion venom sample without sacrifice of the donor animal.

Halophilic microorganisms as a source of novel antibiofilm compounds

Objectives: There is great urgency for alternate sources of antibiotics to be identified. One relatively untapped source of novel bioproducts, including antimicrobials, is organisms derived from extreme environments. Halophiles (which require high salt concentrations) are one such group which is being increasingly explored for their biotechnological potential. The aim of this study was to identify halophilic environmental isolates which possessed in vitro and in vivo antimicrobial and antibiofilm activities. Methods: 73 halophilic bacteria and archaea were isolated from Kilroot salt mine in Northern Ireland. Culture extracts of each isolate were screened for antimicrobial and antibiofilm activity against numerous pathogenic bacteria, including Staphylococcus species and Pseudomonas aeruginosa, both model strains and clinical isolates. The methods used included disc diffusion assays of crude extracts, MIC screening, the MBEC assay, and an in vivo model based on the Greater Wax Moth (Galleria mellonella). Results: The assays indicated >50% of extracts displayed antimicrobial and antibiofilm activity against at least one pathogen, the majority being Staphylococcus species, but also E. coli and P. aeruginosa. Biofilms were either reduced or eradicated by halophile extracts when tested with the MBEC device. Further experiments demonstrated that these effects could be replicated in vivo, with extracts reducing the severity of infections and enhancing the survival of infected G. mellonella. Conclusions: The importance of extremophiles to pharmaceutical research should not be underestimated. While not yet fully characterised, based on the data obtained, the halophiles isolated during this study may provide a promising reservoir of novel antimicrobial and antibiofilm compounds.

Commercial oyster stocks as a potential source of larvae in the regeneration of Ostrea edulis in Strangford Lough, Northern Ireland.

Ostrea edulis was extremely rare in the wild in Strangford Lough from the early 1900s until renewed spatfall was observed at a number of sites in the 1990s. A monitoring programme was undertaken to investigate the presence and distribution of planktonic oyster larvae at nine sites around the lough between June and September in 1997 and 1998 as a precursor to studies of spatfall patterns. Larval densities at sites in the northern basin of the lough were significantly higher than those in the southern basin where larvae were lacking or in low numbers. Densities and sizes of oyster larvae showed significant temporal variation suggesting pulsed larval release. Larval densities also showed significant spatial variation with higher densities at sites closer to commercial stocks pointing to these as the main source of oyster larvae. This hypothesis was supported during a larval flux study over a complete tidal cycle which indicated a 90% net tidal movement of O. edulis larvae from the entrance of the bay where commercial stocks were held to the main body of the lough. Thus the maintenance of dense commercial stocks of flat oysters may provide the key to the redevelopment of native oyster beds in Strangford Lough and elsewhere by providing an initial broodstock nucleus from which larvae can be exported.

Halomethane: bisulfide/halide ion methyltransferase, an unusual corrinoid enzyme of environmental significance isolated from an aerobic methylotroph using chloromethane as the sole carbon source

QUESTOR, DuPont , ICI and EC Framework 4 collaboration (Groningen, Cardiff, Dresden) – Belfast PI Larkin.