Heterologous promoter recognition leading to high-level expression of cloned foreign genes in Bombyx mori cell lines and larvae

The baculovirus expression system using the Autographa californica nuclear polyhedrosis virus (AcNPV) has been extensively utilized for high-level expression of cloned foreign genes, driven by the strong viral promoters of polyhedrin (polh) and p10 encoding genes. A parallel system using Bombyx mori nuclear polyhedrosis virus (BmNPV) is much less exploited because the choice and variety of BmNPV-based transfer vectors are limited. Using a transient expression assay, we have demonstrated here that the heterologous promoters of the very late genes polh and p10 from AcNPV function as efficiently in BmN cells as the BmNPV promoters. The location of the cloned foreign gene with respect to the promoter sequences was critical for achieving the highest levels of expression, following the order +35 > +1 > -3 > -8 nucleotides (nt) with respect to the polh or p10 start codons. We have successfully generated recombinant BmNPV harboring AcNPV promoters by homeologous recombination between AcNPV-based transfer vectors and BmNPV genomic DNA. Infection of BmN cell lines with recombinant BmNPV showed a temporal expression pattern, reaching very high levels in 60-72 h post infection. The recombinant BmNPV harboring the firefly luciferase-encoding gene under the control of AcNPV polh or p10 promoters, on infection of the silkworm larvae led to the synthesis of large quantities of luciferase. Such larvae emanated significant luminiscence instantaneously on administration of the substrate luciferin resulting in 'glowing silkworms'. The virus-infected larvae continued to glow for several hours and revealed the most abundant distribution of virus in the fat bodies. In larval expression also, the highest levels were achieved when the reporter gene was located at +35 nt of the polh.

Linking spatial stock dynamics and economics: Evaluation of indicators and fishery management for the travelling eastern king prawn (Melicertus plebejus)

Reduced economic circumstances havemoved management goals towards higher profit, rather than maximum sustainable yields in several Australian fisheries. The eastern king prawn is one such fishery, for which we have developed new methodology for stock dynamics, calculation of model-based and data-based reference points and management strategy evaluation. The fishery is notable for the northward movement of prawns in eastern Australian waters, from the State jurisdiction of New South Wales to that of Queensland, as they grow to spawning size, so that vessels fishing in the northern deeper waters harvest more large prawns. Bioeconomic fishing data were standardized for calibrating a length-structured spatial operating model. Model simulations identified that reduced boat numbers and fishing effort could improve profitability while retaining viable fishing in each jurisdiction. Simulations also identified catch rate levels that were effective for monitoring in simple within-year effort-control rules. However, favourable performance of catch rate indicators was achieved only when a meaningful upper limit was placed on total allowed fishing effort. Themethods and findings will allow improved measures for monitoring fisheries and inform decision makers on the uncertainty and assumptions affecting economic indicators.

A retrospective evaluation of sustainable yields for Australia's Northern Prawn Fishery

The fundamental aim in fisheries management is to determine an optimal fishing effort for sustainably harvesting from a replenishable resource. The current management objective of Australia's Northern Prawn Fishery is to maximize the long-term net economic return following Australian government policy, resulting in an average recent catch of tiger prawn species of about 1,250 tons only. However, the maximum sustainable catch stated from different studies is around 3,000-4,700 tons. We also evaluated the net profit assuming that there was no buyback scheme in 2005 and the fishing fleet was kept at 89 vessels since 2005 and concluded that 40% more catch on average (2006-2009) and an additional total profit of A$ 17 million ( excluding crew cost) could have been gained in addition to the many millions of dollars of savings in the buyback scheme. These findings have great implications for future management in Australia and elsewhere because there is a grave concern of overfishing worldwide.

Latitudinal and seasonal effects on growth of the Australian eastern king prawn (Melicertus plebejus)

The growth of the Australian eastern king prawn (Melicertus plebejus) is understood in greater detail by quantifying the latitudinal effect. The latitudinal effect is the change in the species' growth rate during migration. Mark-recapture data (N = 1635, latitude 22.21 degrees S-34.00 degrees S) presents northerly movement of the eastern king prawn, with New South Wales prawns showing substantial average movement of 140 km (standard deviation: 176 km) north. A generalized von Bertalanffy growth model framework is used to incorporate the latitudinal effect together with the canonical seasonal effect. Applying this method to eastern king prawn mark-recapture data guarantees consistent estimates for the latitudinal and seasonal effects. For M. plebejus, it was found that growth rate peaks on 25 and 29 January for males and females, respectively; is at a minimum on 27 and 31 July, respectively; and that the shape parameter, k (per year), changes by -0.0236 and -0.0556 every 1 degree of latitude south increase for males and females, respectively.

Tropical prawn trawl bycatch of fish and seasnakes reduced by Yarrow Fisheye Bycatch Reduction Device

Reducing unwanted trawl bycatch is actively encouraged in Australia, particularly in prawn trawl fisheries. We tested the performance of a Bycatch Reduction Device, the Yarrow Fisheye, during two periods of commercial fishing operations in Australia's Northern Prawn Fishery, by comparing the catches of paired treatment and control nets. We compared the catch weights of the small fish and invertebrate bycatch, and the commercially important tiger prawns, from 42 trawls in 2002. The Yarrow Fisheye reduced the weight of small bycatch by a mean of 22.7%, with no loss of tiger prawn. We also compared the numbers of seasnakes caught in 41 and 72 trawls during the spring trawling seasons of 2004 and 2005, respectively. The Yarrow Fisheye reduced the catches by a mean of 43.3%. Flume-tank tests of the Yarrow Fisheye showed that this device created a slow water-flow region extending over 2 m downstream from its position in the net, and close to where the catch accumulates. Finfish and seasnakes may be exploiting this slow water-flow region to escape via the eye, Although the reductions in fish and seasnake bycatch were excellent, we think they could be further improved by relating differences in fisheye position and localised water displacements, to design and rigging changes.

The impact of global positioning systems and plotters on fishing power in the northern prawn fishery, Australia

The impact of global positioning systems (GPS) and plotter systems on the relative fishing power of the northern prawn fishery fleet on tiger prawns (Penaeus esculentus Haswell, 1879, and P. semisulcatus de Haan, 1850) was investigated from commercial catch data. A generalized linear model was used to account for differences in fishing power between boats and changes in prawn abundance. It was found that boats that used a GPS alone had 4% greater fishing power than boats without a CPS. The addition of a plotter raised the power by 7% over boats without the equipment. For each year between the first to third that a fisher has been working with plotters, there is an additional 2 or 3% increase. It appears that when all boats have a GPS and plotter for at least 3 years, the fishing power of the fleet will increase by 12%. Management controls have reduced the efficiency of each boat and lowered the number of days available to fish, but this may not have been sufficient to counteract the increases. Further limits will be needed to maintain the desired levels of mortality.

Stock-recruitment relationships of the tiger prawns (Penaeus esculentus and Penaeus semisulcatus) in the Australian Northern Prawn Fishery

This paper investigates the stock-recruitment and equilibrium yield dynamics for the two species of tiger prawns (Penaeus esculentus and Penaeus semisulcatus) in Australia's most productive prawn fishery: the Northern Prawn Fishery. Commercial trawl logbooks for 1970-93 and research surveys are used to develop population models for these prawns. A population model that incorporates continuous recruitment is developed. Annual spawning stock and recruitment indices are then estimated from the population model. Spawning stock indices represent the abundance of female prawns that are likely to spawn; recruitment indices represent the abundance of all prawns less than a certain size. The relationships between spawning stock and subsequent recruitment (SRR), between recruitment and subsequent spawning stock (RSR), and between recruitment and commercial catch were estimated through maximum-likelihood models that incorporated autoregressive terms. Yield as a function of fishing effort was estimated by constraining to equilibrium the SRR and RSR. The resulting production model was then used to determine maximum sustainable yield (MSY) and its corresponding fishing effort (f(MSY)). Long-term yield estimates for the two tiger prawn species range between 3700 and 5300 t. The fishing effort at present is close to the level that should produce MSY for both species of tiger prawns. However, current landings, recruitment and spawning stock are below the equilibrium values predicted by the models. This may be because of uncertainty in the spawning stock-recruitment relationships, a change in carrying capacity, biased estimates of fishing effort, unreliable catch statistics, or simplistic assumptions about stock structure. Although our predictions of tiger prawn yields are uncertain, management will soon have to consider new measures to counteract the effects of future increases in fishing effort.

Structure of Raster in Melolonthine Larvae

Ultrastructural and electrophysiological investigations carried out on larval rasters of Rhopaea magnicomis Blackburn, Lepidiota frerzclzi Black, and Antitr-ogus consanguineus Blackburn revealed that the raster is a complex of mechanoreceptive setae. Chemical and morphological investigations provide no evidence that the raster is a site for chemical emissions; however, species differences in hydrocarbon profiles were found among larval cuticle samples. Ultrastructure of the setae (pali) show that each seta is innervated by a single dendrite which ends in a tubular body at the base of the seta. The connection with the seta is on the proximal side, which corresponds to the production of a phasic-tonic electrophysiological signal on downward deflection. The dendrite is surrounded by a granular, electron-dense sheath which has inwardly directed arms distally and outwardly directed arms proximally. Two sheath cells are present, 1 forming a large receptor lymph cavity which is lamellate and lined with electron-dense material.

Quantifying the effects of bycatch reduction devices in Queensland's (Australia) shallow water eastern king prawn (Penaeus plebejus) trawl fishery

This study presents results from an experimental 10-day research charter that was designed to quantify the effects of a) a turtle excluder device (TED) and b) a radial escape section bycatch reduction device (BRD) and c) both devices together, on prawn and bycatch catch rates in the Queensland shallow water eastern king prawn (Penaeus plebejus) trawl fishery. The bycatch was comprised of 250 taxa, mainly gurnards, whiting, lizard fish, flathead, dragonets, portunid crabs, turretfish and flounders. The observed mean catch rates of bycatch and marketable eastern king prawns from the standard trawl net (i.e., net with no TED or BRD) used during the charter were 11.06 (se 0.90) kg per hectare swept by the trawl gear (ha-1) and 0.94 kg ha-1, respectively. For the range of depths sampled (20.1-90.7 m), bycatch catch rates declined significantly at a rate of 0.14 kg ha-1 for every 1 m increase in depth, while prawn catch rates were unaffected. When both the TED and radial escape section BRD were used together they resulted in a 24% reduction in total bycatch catch rate compared to a standard net, but at a 20% reduction in marketable prawn catch rate. The largest reductions were achieved for stout whiting Sillago robusta (57% reduction) and yellowtail scad Trachurus novaezelandiae (32% reduction). Multidimensional scaling and analysis of similarities revealed that bycatch assemblages differed significantly between depths and latitude, but not between the different combinations of bycatch reduction devices. Despite the lowered prawn catch rates, the reduced bycatch catch rates are promising, particularly for S. robusta which is not permitted to be retained by the prawn trawl fleet and yet experiences considerable incidental fishing mortality, and because it is targeted in a separate licensed commercial fishery.

Minimising mortality in field and pot experiments using Heteronyx piceus Blanchard (Coleoptera: Scarabaeidae) eggs and larvae

The establishment of experimental populations of scarab larvae using eggs and early instar larvae has proven to be difficult for many researchers. Despite this, little work has been published examining ways to optimise establishment under artificial conditions. In this experiment, we examined the effect of shade and irrigation on the establishment of Heteronyx piceus Blanchard larvae introduced into pots as eggs and first-, second- and third-instar larvae to optimise artificial infestation techniques. The most important factor affecting larval establishment was the life stage introduced. Establishment of eggs and first instars was very low, with only 21% of eggs and 11% of first-instar larvae establishing. In contrast, 82% of second-instar larvae and 84% of third-instar larvae established successfully. The addition of shade marginally improved overall survival from 45% in the unshaded pots to 53% in the shaded pots. However, most of this increase was in the eggs and first instars. Irrigation did not improve survival. These results suggest that when introducing scarab larvae to field or pot experiments, second- or thirdinstar larvae should be used to maximise establishment. The provision of shade and supplementary irrigation is optional.

Effect of soil burial depth and wetting on mortality of diapausing larvae and patterns of post-diapause adult emergence of sorghum midge, Stenodiplosis sorghicola (Coquillett) (Diptera: Cecidomyiidae)

In south-eastern Queensland, Australia, sorghum planted in early spring usually escapes sorghum midge, Stenodiplosis sorghicola, attack. Experiments were conducted to better understand the role of winter diapause in the population dynamics of this pest. Emergence patterns of adult midge from diapausing larvae on the soil surface and at various depths were investigated during spring to autumn of 1987/88–1989/90. From 1987/88 to 1989/90, 89%, 65% and 98% of adult emergence, respectively, occurred during November and December. Adult emergence from larvae diapausing on the soil surface was severely reduced due to high mortality attributed to surface soil temperatures in excess of 40°C, with much of this mortality occurring between mid-September and mid-October. Emergence of adults from the soil surface was considerably delayed in the 1988/89 season compared with larvae buried at 5 or 10 cm which had similar emergence patterns for all three seasons. In 1989/90, when a 1-cm-deep treatment was included, there was a 392% increase in adult emergence from this treatment compared with deeper treatments. Some diapausing larvae on the surface did not emerge at the end of summer in only 1 year (1989/90), when 28.0% of the larvae on the surface remained in diapause, whereas only 0.8% of the buried larvae remained in diapause. We conclude that the pattern of emergence explains why spring plantings of sorghum in south-eastern Queensland usually escape sorghum midge attack.

Aspects of reproduction and diet of the Australian endemic skate Dipturus polyommata (Ogilby) (Elasmobranchii: Rajidae), by-catch of a commercial prawn trawl fishery

The Australian endemic skate Dipturus polyommata collected from by-catch of a benthic prawn fishery off southern Queensland was examined to provide information on reproduction and diet. Morphological relationships of total length (LT) to disc width and LT to mass were estimated. Size at birth was estimated at c. 100-110 mm and size at first feeding at c. 105-110 mm LT. Size at 50% maturity (LT50 and 95% CI) was 321 (305-332) and 300 (285-306) mm LT for females and males, respectively. Size at first maturity corresponded to 87.7% of observed maximum size in females (366 mm LT) and 87.5% in males (343 mm L T). Two females, representing 18.2% of mature females sampled in the austral winter were each carrying two egg cases. Descriptions of egg cases are given. Diet described by the index of relative importance as a percentage (%IRI) was predominantly crustacean based with carid shrimps (53.64%) and penaeoid prawns (23.30%) the most significant prey groups. Teleosts (11.72%), gammarid amphipods (5.31%) and mysids (4.72%) were also important to the diet of the species, while a further six prey groups made only a minor contribution to diet (1.31%). An ontogenetic change was evident between the diets of immature and mature skates. Immature animals fed more extensively on carids and amphipods and mature animals on penaeoids, teleosts and mysids.

Developments in controlled green-water larval culture technologies for estuarine fishes in Queensland, Australia and elsewhere

Since 1989, researchers with the Department of Primary Industries and Fisheries (DPI&F) in Queensland, Australia, have successfully used controlled low-water exchange green-water cultures to rear the larvae of estuarine fishes and crustaceans through to metamorphosis. High survivals and excellent fry condition have been achieved for several commercially important endemic species produced for various projects. They include barramundi or sea bass, Lates calcarifer, Australian bass, Macquaria novemaculeata, dusky flathead, Platycephalus fuscus, sand whiting, Sillago ciliata, red sea bream or snapper, Pagrus auratus, banana prawn, Fenneropenaeus merguiensis, and others. The consistent success of our standardised and relatively simple approach at different localities has led to it being incorporated into general fingerling production practices at several establishments in Australia. Although post-metamorphosis rearing methods have differed for each species investigated, due to various biological and behavioural traits and project requirements, these larval rearing methods have been successful with few species-specific modifications. Initially modelled on the Taiwanese approach to rearing Penaeids in aerated low-water exchange cultures, the approach similarly appears to rely on a beneficial assemblage of micro-organisms. Conceptually, these micro-organisms may include a mixture of the air-borne primary invaders of pure phytoplankton cultures when exposed to outdoor conditions. Whilst this would vary with different sites, our experiences with these methods have consistently been favourable. Mass microalgal cultures with eco-physiological youth are used to regularly augment larval fish cultures so that rearing conditions simulate an exponential growth-phase microalgal bloom. Moderate to heavy aeration prevents settlement of particulate matter and encourages aerobic bacterial decomposition of wastes. The green-water larval rearing approach described herein has demonstrated high practical utility in research and commercial applications, and has greatly simplified marine finfish hatchery operations whilst generally lifting production capacities for metamorphosed fry in Australia. Its potential uses in areas of aquaculture other than larviculture are also discussed.

Survival of Campylobacter spp. in darkling beetles (Alphitobius diaperinus) and their larvae in Australia

Campylobacter infection is the most frequently reported notifiable food-borne disease in humans in Australia. Our studies investigated the persistence of Campylobacter spp. in or on darkling beetles (Alphitobius diaperinus) and their larvae. Our results in analyses with chickens confirm that, unless very short turnaround times are used (<72 h), beetles colonized in one production cycle (i.e., one batch of chickens) are most unlikely to still be colonized during the next cycle of chickens.

Population genetic structure of the brown tiger prawn, Penaeus esculentus, in tropical northern Australia

Eight polymorphic microsatellite loci were analysed in six population samples from four locations of the Australian endemic brown tiger prawn, Penaeus esculentus. Tests of Hardy-Weinberg equilibrium were generally in accord with expectations, with only one locus, in two samples, showing significant deviations. Three samples were taken in different years from the Exmouth Gulf. These showed no significant heterogeneity, and it was concluded that they were from a single panmictic population. A sample from Shark Bay, also on the west coast of Australia, showed barely detectable differentiation from Exmouth Gulf (F (ST) = 0 to 0.0014). A northeast sample from the Gulf of Carpentaria showed low (F (ST) = 0.008) but significant differentiation from Moreton Bay, on the east coast. However, Exmouth Gulf/Shark Bay samples were well differentiated from the Gulf of Carpentaria/Moreton Bay (F (ST) = 0.047-0.063). The data do not fit a simple isolation by distance model. It is postulated that the east-west differentiation largely reflects the isolation of east and west coast populations that occurred at the last glacial maximum when there was a land bridge between north-eastern Australia and New Guinea.