Comparisons of physical separation methods of Kunjin virus-induced membranes

The two sets of connected membranes induced in Kunjin virus-infected cells are characterized by the presence of NS3 helicase/protease in both, and by RNA-dependent RNA polymerase (RdRp) activity plus the associated double-stranded RNA (dsRNA) template in vesicle packets (VP), or by the absence of both the VP-specific markers in the convoluted membranes/paracrystalline arrays (CM/PC). Attempts were made to separate flavivirus-induced membranes by sedimentation or flotation analyses in density gradients of sucrose or iodixanol, respectively, after treatment of cell lysates by sonication, osmotic shock, or tryptic digestion. Only osmotic shock treatment provided suggestive evidence of separation. This was explored by flow cytometry analysis (FCA) of RdRp active membrane fractions from a sucrose gradient, using dual fluorescent labelling via antibodies to NS3 and dsRNA. FCA revealed the presence of a dual labelled membrane population indicative of VP, and in a faster sedimenting fraction a membrane population able to be labelled only in NS3, representative of CM/PC and associated (R)ER. It was postulated that osmotic shock ruptured the bounding membrane of the VP, releasing the enclosed small vesicles associated with the Kunjin virus replication complex characterized previously. Notably, the presence of the full spectrum of nonstructural proteins in some membrane fractions was not a reliable marker for RdRp activity. These experiments may provide the opportunity for isolation of relatively pure flavivirus replication complexes in their native membrane-associated state by fluorescence-activated cell sorting. (C) 2004 Elsevier B.V. All rights reserved.

Germination response of Phalaris paradoxa L. seed to different light qualities

The influence of different light regimes on the germination of Australian and English populations of Phalaris paradoxa L. (awned canary-grass) seed was investigated to determine the impact of changing tillage practices on weed infestation. Seeds of all biotypes were highly viable, but differed in levels of innate dormancy (26-99%). In one experiment seed from a single Australian biotype, either enclosed in the spikelet glumes or having the spikelet glumes removed, were exposed to nine light treatments. Germination was stimulated by red and white light, but was inhibited by far-red light. Time to 50%, germination was less for seed enclosed in the spikelet glumes than for naked caryopses, although the final percentage of seed germinating when still enclosed in the spikelet glumes was significantly lower than for naked caryopses. In another experiment, six Australian and English biotypes with varying dormancy characteristics were exposed to eight light treatments. Red light did not stimulate germination in the deeply dormant biotype, however stimulated all other biotypes. Germination in darkness was below 20% in all biotypes except for one where germination was 51%. To overcome dormancy seeds were imbibed and placed in darkness at 16degreesC for either 7 or 14 days prior to exposure to red or white light for a single 15-min period. Dormancy in all biotypes was overcome indicating that a period of burial may decrease the dormancy level and increase seed sensitivity to light. This increased light sensitivity suggests that exposure to light during tillage may stimulate germination in P. paradoxa seed.

Anatomy of ethylene-induced floral-organ abscission in Chamelaucium uncinatum (Myrtaceae)

Postharvest abscission of Geraldton waxflower (Chamelaucium uncinatum Schauer) flower buds and flowers is ethylene-mediated. Exposure of floral organs to exogenous ethylene (1 mu L L-1) for 6 h at 20 degrees C induced separation at a morphologically and anatomically distinct abscission zone between the pedicel and. oral tube. Flower buds with opening petals and flowers with a nectiferous hypanthium were generally more responsive to exogenous ethylene than were flower buds enclosed in shiny bracteoles and aged (senescing) flowers. The anatomy of abscission-zone cells did not change at sequential stages of floral development from immature buds to aged flowers. The zone comprised a layer of small, laterally elongated-to-rounded, closely packed and highly protoplasmic parenchyma cells. Abscission occurred at a two- to four-cell-wide separation layer within the abscission zone. The process involved degradation of the middle lamella between separation layer cells. Following abscission, cells on both the proximal and distal faces of the separation layer became spherical, loosely packed and contained degenerating protoplasm. Central vascular tissues within the surrounding band of separation layer cells became torn and fractured. For flower buds, bracteoles that enclose the immature floral tube also separated at an abscission zone. However, this secondary abscission zone appeared less sensitive to ethylene than the primary ( central). oral-tube abscission zone as bracteoles generally only completely abscised when exposed to 10 mu L L-1 ethylene for the longer period of 24 h at 20 degrees C. The smooth surfaces of abscised separation-layer cells suggest that hydrolase enzymes degrade the middle lamella between adjacent cell walls.

Atoll lagoon flushing forced by waves

Water level and current measurements from two virtually enclosed South Pacific atolls, Manihiki and Rakahanga, support a new lagoon flushing mechanism which is driven by waves and modulated by the ocean tide for virtually enclosed atolls. This is evident because the lagoon water level remains above the ocean at all tidal phases (i.e., ruling out tidal flushing) and because the average lagoon water level rises significantly during periods with large waves. Hence, we develop a model by which the lagoons are flushed by waves pumping of ocean water into the lagoon and gravity draining water from the lagoon over the reef rim. That is, the waves on the exposed side push water into the lagoon during most of the tidal cycle while water leaves the lagoon on the protected side for most of the tidal cycle. This wave-driven through flow flushing is shown to be more efficient than alternating tidal flushing with respect to water renewal. Improved water quality should therefore be sought through enhancement of the natural wave pumping rather than by blasting deep channels which would change the system to an alternating tide-driven one.

Using machine vision classification to control access of animals to water

Invasive vertebrate pests together with overabundant native species cause significant economic and environmental damage in the Australian rangelands. Access to artificial watering points, created for the pastoral industry, has been a major factor in the spread and survival of these pests. Existing methods of controlling watering points are mechanical and cannot discriminate between target species. This paper describes an intelligent system of controlling watering points based on machine vision technology. Initial test results clearly demonstrate proof of concept for machine vision in this application. These initial experiments were carried out as part of a 3-year project using machine vision software to manage all large vertebrates in the Australian rangelands. Concurrent work is testing the use of automated gates and innovative laneway and enclosure design. The system will have application in any habitat throughout the world where a resource is limited and can be enclosed for the management of livestock or wildlife.

Effectiveness of evaporative cooling of beef cattle housed in confinement

Eighteen Angus steers exposed to high heat load conditions were used to assess the effectiveness of four spray cooling systems, on reducing the effects of heat load, the impact on microclimate and water usage. The steers were housed in groups of nine in a fully enclosed shed and were exposed to high heat load conditions for four days. The cooling systems used were water applied via a hose, via overhead sprinklers, via sprinklers at leg height and via misters. The water used was approximately 31 oC and contained 3% NaCl. Fans were used to ensure adequate air movement over the cattle. The animal parameters measured were feed intake, respiration rate, panting score and behaviour. Climatic factors were ambient temperature and wet bulb temperature. Ammonia levels were also measured. The hose, overhead sprinklers and misting were successful in reducing heat load on the cattle. The leg wetting system did not work because the dominant cattle blocked access to the sprinklers. The misting system used the most water (5483 L) and the hose the least (845 L). The application of water had minor impacts on wet bulb temperature, but resulted in significant reductions in dry bulb temperatures.

Holocene climate records in coral reefs from the South China Sea

Coral reefs, excellent climatic and environmental archives in tropical oceans, are widely distributed in the South China Sea (SCS), which is the largest enclosed marginal sea of western Pacific, covering over 20° in latitude and different climate conditions. Our recent research in the SCS focuses on coral-based high-resolution climate reconstruction and coral reef ecological responses using geochemical and U-series geochronological tools, which provide an ideal opportunity for understanding of Holocene climate processes and events. Some major research highlights are summarized below: