Reappearance of Eumarcia fumigata (G.B. Sowerby II, 1853) (Bivalvia: Veneridae) into the Swan River,Western Australia

ABSTRACT

The venerid bivalve Eumarcia fumigata was a common species in Western Australia (WA) during the Pleistocene, where it was distributed as far north as Shark Bay. It became extinct in WA as the climate changed several thousand years ago but remains common in eastern Australia from southern Queensland to South Australia and Tasmania. The species has recently reappeared in the Swan River, probably due to shipping movements. Of the > 60 marine and estuarine species introduced into WA it is only the third confirmed introduction from eastern Australia, and the first that is a reappearance in the Swan River of a species present in the geological past. The present reappearance of E. fumigata, and the introduction of other species, has been made possible by the removal of a rocky bar at the mouth of the estuary and the creation of more marine conditions in the lower Swan estuary.     

Improved sensitivity,accuracy and prediction provided by a high-performance liquid chromatography screen for the isolation of phytase-harbouring organisms from environmental samples

HPLC methods are shown to be of predictive value for classification of phytase activity of aggregate microbial communities and pure cultures. Applied in initial screens, they obviate the problems of ‘false-positive’ detection arising from impurity of substrate and imprecision of methodologies that rely on phytate-specific media. In doing so, they simplify selection of candidates for biotechnological applications. Combined with 16S sequencing and simple bioinformatics, they reveal diversity of the histidine phosphatase class of phytases most commonly exploited for biotechnological use. They reveal contribution of multiple inositol-polyphosphate phosphatase (MINPP) activity to aggregate soil phytase activity, and they identity Acinetobacter spp. as harbouring this prevalent soil phytase activity. Previously, among bacteria MINPP was described exclusively as an activity of gut commensals. HPLC methods have also identified, in a facile manner, a known commercially successful histidine (acid) phosphatase enzyme. The methods described afford opportunity for isolation of phytases for biotechnological use from other environments. They reveal the position of attack on phytate by diverse histidine phosphatases, something that other methods lack.     

Purification and partial characterisation of a broad-range L-amino acid oxidase from Bacillus carotarum 2Pfa isolated from soil

The L-amino acid oxidase (L-aao) from Bacillus carotarum 2Pfa was purified to homogeneity, as judged by polyacrylamide gel electrophoresis, from crude sonicated cell extract by a combination of anion exchange chromatography and gel filtration. The purified enzyme was a dimer with a native relative molecular mass of approximately 102,000 to 115,000 and comprised two identical subunits of 54,000. The isoelectric point of the L-aao was at pH 4.8 the ph optimum was at 8.0–8.5 and the temperature optimum was at approximately 50° C. It was stable for several months at + 4° C and at –20° C. The enzyme contained 2 mol flavin adenine dinucleotide (FAD)/mol enzyme and exhibited relatively broad range substrate specificity, oxidising a total of ten L-amino acids and , albeit to a much lesser extent, seven D-amino acids. Kinetic studies revealed that the three aromatic L-amino acids were the preferred substrates.     

How does light and phosphorus fertilisation affect the growth and ectomycorrhizal community of two contrasting dipterocarp species?

Phosphorus concentrations in many south-east Asian tropical rain forest soils are very low. To determine the growth responses of seedlings of a light-demanding (Shorea leprosula) and a more shade-tolerant (Hopea nervosa) dipterocarp species to increasing P, we carried out a nursery fertilisation experiment. Responses of symbiotic ectomycorrhizal (EcM) fungi to the treatments were also determined. Seedlings were grown under high light (13 mol m⁻² d⁻¹) or moderate light (4 mol m⁻² d⁻¹) in shade-chambers and were fertilised with a solution containing 0, 1, 10 or 100 mg L⁻¹ P. The growth of Hopea and Shorea showed different responses to the light and P fertilisation treatments with Hopea having greater growth under moderate light conditions and Shorea having greater growth under high light conditions. Shorea responded to P fertilisation by increasing its foliar P concentrations and growth rates, whereas Hopea did not take up additional P and did not improve its growth rates. There was no effect of either light or P fertilisation on total EcM colonisation or EcM diversity, but around half of the EcM morphotypes observed were affected by one of these two abiotic perturbations, most notably for Riessiella sp. which increased with P fertilisation suggesting it may not be a mutualistic fungus. These results show how niche partitioning in both dipterocarp seedlings and EcM fungi can be divided along contrasting axes.     

Response of the porcine MYH4-promoter and MYH4-expressing myotubes to known anabolic and catabolic agents in vitro

Myosin heavy chain-IIB (MyHC-IIB; encoded by MYH4 or Myh4) expression is often associated with muscle hypertrophic growth. Unlike other large mammals, domestic pig breeds express MyHC-IIB at both the mRNA and protein level.AimTo utilise a fluorescence-based promoter-reporter system to test the influence of anabolic and catabolic agents on increasing porcine MYH4-promoter activity and determine whether cell hypertrophy was subsequently induced.MethodsC2C12 myoblasts were co-transfected with porcine MYH4-promoter-driven ZsGreen and CMV-driven DsRed expression plasmids. At the onset of differentiation, treatments (dibutyryl cyclic-AMP (dbcAMP), Des(1–3) Insulin-Like Growth Factor-1 (IGF-I), triiodo-l-thyronine (T3) and dexamethasone (Dex)) or appropriate vehicle controls were added and cells maintained for up to four days. At day 4 of differentiation, measurements were collected for total fluorescence and average myotube diameter, as indicators of MYH4-promoter activity and cell hypertrophy respectively.ResultsPorcine MYH4-promoter activity increased during C2C12 myogenic differentiation, with a marked increase between days 3 and 4. MYH4-promoter activity was further increased following four days of dbcAMP treatment and average myotube diameter was significantly increased by dbcAMP. Porcine MYH4-promoter activity also tended to be increased by T3 treatment, but there were no effects of Des(1–3) IGF-I or Dex treatment, whereas average myotube diameter was increased by Des(1–3) IGF-I, but not T3 or Dex.ConclusionPorcine MYH4-promoter activity responded to dbcAMP, Des(1–3) IGF-I and T3 treatment in vitro as observed previously in reported in vivo studies. However, we report that increased MYH4-promoter activity was not always associated with muscle cell hypertrophy. The fluorescence-based reporter system offers a useful tool to study muscle cell hypertrophic growth.